TRENDS IN BIOSCIENCES JOURNAL 6-2 JUNE 2013 EDITION

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Trends 

in 

Biosciences 


Volume 6 Number 2 

April, 2013 



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Trends in Biosciences 

Volume 6 Number 2 April, 2013 

CONTENTS 

MINI REVIEW 

1. Mechanism of Cadmium Induced Hepatocellular Carcinoma 125 

Sundas Asghar, Muhammad Saif Ur Rahman and Usman Ali Asfaq 

2. Role of Herbal Ice Cream in Human Health: A Review 130 

Swashankh Kumar, D.C. Rai and Dinker Singh 

3. Management of Stored-product Insect Pests Through Biorational Approaches: A review 133 

S. Murali 

RESEARCH PAPERS 

4. Preliminary Studies of Phenology of Some Selected Tree Species from Ngel-Nyaki Forest reserve 138 

Mohammed, S., Aliyu, B., Umar, M.D. 

5. In Vitro Evaluation of Antibacterial Activity of A Novel Polyenzyme Preparation Immunoseb 142 

Anita Joshi, Varsha Shahane, Varsha Gore, Anju Kagal, Shilpa Risbud and Renu Bharadwaj 

6. Comparative Analysis of Morphological and Molecular Diversity in Mungbean (Vigna radiata L. Wilczek) 146 

G. Roopa Lavanya and Shirish A. Ranade 

7. Efficient Protocol for In Vitro Regeneration in Pigeonpea (Cajanus cajan (L.) Millsp.) 152 

Tushita Gargi, S. Acharya, J.B. Patel and Vandana Thakkar 

8. Study on Two Edible Spiders of the Genus: Nephila (Fam. Nephilidae) of Manipur, India 154 

A.Kananbala, M. Bhubaneshwari and Manju Siliwal 

9. Effect of Coagulants on Nutrient and Antinutrient Parameters of Soy Tofu 158 

M.K.Tripathi, S.mangaraj 

10. Integrated Weed Management Studies on Weed Flora and Yield in Kharif Maize 161 

Birendra Kumar, Ranvir Kumar, Suman Kalyani and Mizzanul Haque 

11. Influence of Pseudomonas putida on the Yield of Agaricus bisporus (Lange) Imbach 165 

Prabhat Kumar Singh, Abhilasha A. Lal , Sobita Simon and Satish Sharma 

12. Effect of Varying NPK Levels and Bio-fertilizers on Growth and Yield of Okra 

[Abelmoschus esculentus (L.) Moench] under Sustainable Condition 167 

Param Hans Prasad and Abhishek Naik 

13. Breeding Periodicity of the Mullet, Liza macrolepis from Mangalore Waters 170 

H.N. Anjanayappa1, S. Benakappa, S., M. Shivaprakash, S.R. Somashekara, A. S. Kumar Naik, 

Jitendra Kumar, Mahesh, V. 

14. Effect of Biofertilzers on Growth and Yield Attributes of Pea (Pisum sativum L.) 174 

Pushkar Singh Patel, R.B. Ram , Jayprakash and M.L.Meena 

15. Studies of Quality Characteristics in Short Grain Scented Rice (Oryza sativa L.) Varieties accessions 177 

Sunita Kumari1, R.N.Kewat, R.P.Singh and Pratibha Singh 

16. Studies on the Association of Plant Parasitic Nematodes Associated with Root-knot Nematode Infecting 

Potato (Solanum tuberosum) 180 

Zenith N.G., Joymati L and Ronibala K.H. 

17. Taxonomic Study on Fishes in the Rivers of Imphal Valley 182 

N. Mohendra Singh 

18. Analyses of the Forest Cover Change in Rani and Garbhanga Reserve forest, Assam, North East 

India Using Geospatial Technique 185 

H Suchitra Devi, A. Pinokiyo1 and S.K. Borthakur 

19. Effect of Different Levels of Boron and Sulphur on Growth of Chickpea with Mustard Intercropping System 188 

Sunil Kumar, S.K. Patel and Gautam Ghosh

20. Efficacy of Fungicides on In Vitro Growth of Pigeonpea against Stem canker 190 

Srujani Behera, R. B. Singh and Laxman Prasad Balai 

21. Influence of Microbial Inoculants and Nutrients on Morpho-Physiological, Growth Parameters and 

Yield Potential in Tomato (Lycopersicon esculentum L. Mill.) 192 

Mohan Kumar, K.G., and Chetti M.B. 

22. Sensory Quality Evaluation of MA Packaged Fruits Applying Fuzzy Logic 195 

S. Mangaraj, M.K. Tripathi 

23. Effect of Menopause on Serum Lipid Profile Pattern in Women 200 

Ekta A.Andriyas, Sapna Smith Lal 

24. Sensory Evaluation of Vegetables Grown Under Organic and Inorganic Conditions 203 

Bajpai Preeti and Punia Darshan 

25. Hypolipidemic Potential of Bacopa monniera in Cholesterol Fed Rats 206 

Syed Mansoor Ali and Gyan Chand Jain 

26. Chemoattraction in Entomopathogenic Nematodes 210 

Rashid Pervez and S. S. Ali 

27. Effectiveness and Economics of Integrated Weed Management in Transplanted Rice (Oryza sativa) 212 

Birendra Kumar, Ranvir Kumar, Suman Kalyani and M. Haque 

28. Biology of Mango - Hopper, Amritodus atkinsoni (Leth.) (Jassidae : Hemiptera) in Agro - ecosystem of Manipur 216 

M. Bhubaneshwari Devi 

SHORT COMMUNICATIONS 

29. A New Blight Disease of Rice Caused by Curvularia sp. from U.P. 218 

Kamaluddeen, Sobita Simon and Abhilasha A. Lal 

30. Exploring Precision Agriculture Approaches for Insect Pest Management 219 

Prashant Kumar and Ashutosh Kumar 

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Trends in Biosciences 6 (2): 125-129, 2013 


Mechanism of Cadmium Induced Hepatocellular Carcinoma 

SUNDAS ASGHAR, MUHAMMAD SAIFUR RAHMAN AND USMAN ALI ASFAQ 

Govt. College University Faisalabad, Allama Iqbal Road, Faisalabad Pakistan 38000. 

e-mail: sundaskth@gmail.com 

ABSTRACT 

Cadmium (Cd) is an important environment pollutant having 

carcinogenic properties. This heavy metal poses serious threats 

to the animals and human beings. From the environment it 

enters into the animals and human bodies and destroyed the 

basic organs of the body including lungs, pancreas, testis, liver 

and kidney. Liver is the basic primary organs involved in the 

elimination of toxic effects and removal of wastes from the 

body. This organ is more prone to the adverse effects of this 

metal than any other organ. Cd causes oxidative stress and 

apoptosis which eventually lead to hepatocellular dysfunction. 

The review is to highlight the current data available regarding 

the Cd induced cellular and molecular mechanisms involved 

in apoptosis, necrosis and hepatocellular carcinoma. 

Key words Cadmium, Hepatotoxicity, Hepatocytes, Mitochondria, 

Metallothionien, Oxidative Stress, Apoptosis, Hepatocellular 

carcinoma. 

Cd is ranked seventh in the Top 20 Hazardous substance 

priority list published by U.S. Environmental Protection 

Agency and “Agency for Toxic Substances and Disease 

Registry” (Fay and Mumtaz, 1996). International agency of 

research on cancer (IARC) has classified Cd as the category 

1 carcinogen. Cadmium(Cd) enters in the environment by 

various human activities or natural activities e.g. volcanic 

activity, erosion and weathering,river transport, tobacco 

smoke,smelting, mining, non-ferrous metals. Moreover, 

municipal wastes, Fossil fuels combustion, refining, plastic 

wastes, Cd containing batteries, fertilizers manufacturers, 

Cd plated steel scrap recycling electrical and electronic waste 

are also important sources of Cd induced toxicity (UNEP 2008). 

Nanotechnology having Cd containing non particles can also 

be source of Cd accumulation. 

General Effect of Cadmium to the Human Health : 

Cd is toxic and non-essential but is taken into the cell by 

the process termed as ionic and molecular mimicry. Cd has no 

known beneficial effects in humans. Individuals exposed to 

Cd can show serious effects such as pulmonary edema, renal 

and hepatic dysfunction, osteomalacia, testicular damage 

(IARC, 1993). Cd effects number of tissues and organs in 

humans including lungs, pancreas, testis, placenta, bone, 

kidney and liver (Zalups and Ahmad, 2003). Cd caused 

oxidative DNA damage, DNA protein cross linkage, inhibition 

of DNA repair and apoptosis (Filipic and Hei, 2004). Cd toxicity 

Sundas Asghar 

Sundas Asghar completed MS in Molecular 

Biology/Biomedicine from University of Skövde 

(HIS) Sweden, with research projects in 

“Investigation on gene regulation and signal 

transduction in cadmium induced carcinogenesis” and 

“An in vitro cell system based approach to clarify 

the specific molecular mechanisms behind the proposed estrogenic 

effects of cadmium in estrogen responsive tissues/cells” from 

Department of Environmental Medicines (IMM), Karolinska 

Institute, Stockholm, Sweden. She was haveing working experience 

as researcher with project in “Molecular genetics of sediment and 

C. metallidurans (CH3)” at Laboratory of Applied Microbiology- 

Bioengineering (GEBI), Earth and Life Institute (ELIM), UCL, 

Belgium. Presently working as lecturer in Department of 

Bioinformatics and Biotechnology at Government College 

University, Faisalabad, Pakistan. Presently she is working 

experience on effects of heavy metals in fish, as well as on aquatic 

toxicity and health, health risk assessments of environmental 

carcinogens and molecular biology of environmental toxicants 

leading to carcinogenesis. 

Usman Ali Ashfaq 

Dr. Usman Ali Ashfaq has done Research on 

“Studies on the therapeutic effect of selected 

phytochemicals against Hepatitis C Virus” at Center 

of Excellence in Molecular Biology, University of the 

Punjab, Lahore. He has worked at a Center of 

Excellence in Molecular Biology, University of the 

Punjab also post Ph.D. Research Experience He has worked at 

Allama Iqbal Medical College and Research center. Presently, he is 

working as Assistant Professor at Department of Bioinformatics 

and Biotechnology at Government College University, Faisalabad. 

Dr. Asfaq also worked on antiviral drugs against HCV, siRNAs 

against viral and cellular genes. He has 28 research paper to his 

credit published and foreign journals. 

Muhammad Saifur Rahman 

Mohd. Saifun Rahman, doing research project 

on “Investigating the cadmium induced hepatocellular 

and nephrotoxic effects in mice” at Department of 

Bioinfomatics and Biotechnology, Government 

College University Faisalabad, Pakistan. Earlier he 

was engaged on research progress of “Biodiversity of snails from 

the agro-ecosystem of tehsil Faisalabad city”. He was working 

experience on projects in “Heavy metal toxicity in fish” and 

“Toxicity of various insecticides on various developmental stages 

of Trichogramma” at Govt. College University Faisalabad, Pakistan. 

He is co-author of two books namely 

1. Toxic effects of different insecticides on Trichogramma 

cholini. 

2. Blood and renal pathology in labeo rohita exposed to 

mercuric chloride.

126 Trends in Biosciences 6 (2), 2013 

increases the negative effects on the anti-oxidants defense 

system. Cd and Cd-Mg causes oxidative stress in liver 

(Matovic, et. al., 2012). Cd is very toxic and harmful pollutant 

and bio-accumulation in various levels of food chain. Cd 

exposure damages various organs of the body especially in 

liver and kidney (Jihen, et. al., 2009). 

Damage to Liver : 

Hepatic Response : 

Cd either taken orally or via pulmonary route or parental 

exposure, liver is the primary organ that encounters with the 

excess level of Cd in the early hours after the exposure as 

shown in Fig 1. It has been investigated that Cd induces 

hepatotoxicity injury and ischemia due to endothelial cells 

injury by the activation of inflammatory cytokines (Yamano, 

et al., 2000). Cd exposure results in severe glycogen depletion, 

cellular degradation necrosis and other effects to the liver i.e. 

loss of normal architecture of the parenchymatous tissues, 

cytoplasm vacuolization, cellular degeneration, necrosis, 

congested blood vessels, destructed Mitochondrial cristae, 

fat globules, severe glycogen depletion, lipofuscin pigments 

and collagenous fibers formation are observed in liver tissues 

on exposure to Cd (El-Sokkary, et. al., 2010). These cellular 

changes result in necrosis and apotosis. Hepatic injury is 

appeared to be caused by the association of sulfhydral groups, 

membrane and cytoplasmic proteins and enzymes. Cd is non 

redox metal and can deplete the cellular level of glutathione 

which can induce oxidative stress(Martelli, et. al., 2006). Cd 

interfering with cellular processes that are energy metabolism 

and metal membrane transport. Cd affectsvarious cellular 

functions, enzyme activities; redox state of cells; signal 

transduction and DNA repair system (Van, et. al., 2010). It 

disassembles E-Cadherin/beta-catenin complex and changes 

cell to cell adhesion. Cd affects the translational machinery in 

liver cells. Cd shows the increased; DNA damage, level of 

lipid per oxidation in liver, apoptosis and altered histology. 

Cd effects the metallothionein gene expression in liver. It has 

greater interaction with Mt-mRNA and proteins as compared 

to other heavy metals (Zhang, et. al., 2012). 

Cd infiltrated by polymorphonuclear neutrophil 

(PMN)and Kupffer cells causes hepatotoxicity by enhancing 

the necrosis and promoting inflammation in the liver 

(Horiguchi, et. al., 2000). Variety of cytotoxic mediator i.e.nitric 

oxide, cytokines and reactive oxygen species (ROS) are 

activated by Kupffer cellswhich damage thehepatocytes 

directly as shown in Fig 2. Tumor necroses factor also plays 

animportantrole in Cd toxicity (Mousa, 2004). It is thought 

that for the Cd induced hepatotoxicitythe hepatic endothelial 

cells might be the first cellular target. Cd induced hepatic 

endothelial degeneration was observed after the early hours 

of exposure. 

Damaged liver cells enter into Capillary lumen which 

leads to Ischemia(Rikans and Yamano, 2000). The injured 

endothelial cells impede the capillary lumen and develop 

ischemia which in turn may induct molecular and cellular 

events accompanied by the activation of Kupffer cells, bring 

out inflammatory mediator and enlisting of inflammatory cells 

chiefly PMN and leukocytes. In liver inflammatory cells 

cumulate in sinusoidal andcohere to endothelial cells. This is 

facilitated by the E-selectin, Mac-1 and ICAM-1. Plateletendothelial 

adhesion molecule-1 (PECAM-1) has an implicative 

role during hepatotoxicity in adhesion molecule (Mousa, 2004) 

as shown in Fig 2. ROS also enhance inflammation by 

activating the transcription factors i.e.; activator protein-1 

(AP-1) and nuclear factor-kB (NF-kB) which caters signals 

responsible for the expression of adhesion molecule and 

pro-inflammatory genes (Jaeschke, 2000; Souza, et. al., 2004). 

Fig. 1. 

Cd entres into the body by the following routes. Cd 

through food enters into GIT from where it enters into 

the Liver through blood. Cd enters into Blood and then 

to liver through skin and through lungs via respiratory 

inhalation. 

Fig. 2. 

It is shown that how Cd indduces inflammatory 

response in Kupffer cells cells which activate ROS and 

Cytokines damage hepatocytes PMN also effected by 

inflammatory response and affirms.

ASGHAR et. al.,: Mechanism of Cadmium Induced Hepatocellular Carcinoma 127 

Cd and cellular Cd traffic deregulates the metal 

homeostasis and thus contributes to Cd toxicity. Cd and Zn 

has same chemical group and do not change their oxidation 

state. In biological environment they are in the form of cations. 

Cd in metalloprotien binds with sulfur exclusively but Cd also 

binds with Zn in coordination in alcohol dehydrogenase site. 

This is also explained as the molecular mechanism responsible 

for Cd hepatotoxicity (Moulis, 2010). 

MT is involved in Zn exchange between proteins ad Zn 

buffering. Cd bound tightly with MT and produce stress which 

in result mediate Cd toxicity because of highly sensitive 

expression of MT genes under stress. Improper expression 

decreases the ability of Zn buffering by decreasing the 

antioxidant cellular defense and changing the Zn exchange. 

Cd accumulation can destroy Zn homeostasis and can increase 

hepatic Zn by redistributing the Zn from non hepatic tissues 

by increasing hepatic and aminolevulinate dehydratase (and 

ALA-D). Cd is non essential metal and it is assumed that it is 

transported by the transport system of essential metals Zn, 

Fe, Ca, Mn and Mg transporters are involved in the transport 

of Cd to liver cells (Zhang, et. al., 2001). 

Cd has almost the same ionic radius as that of Ca which 

is helpful in the transport of cadmium. Cadmium replaces Ca 

and binds with the calcium bindingproteins and via these Ca 

channels Cd makes its way into the cells. After entrance in 

hepatocytes Cd alters the intracellular concentration of Ca by 

the mobilization of Ca from its storage site in the cells. Cd 

modulate extracellular calcium sensing receptor (CaSR), CaSR 

proteins are G proteins-coupled receptors which express in 

hepatocytes by this mechanism Cd interferes with Ca 

Homeostasis (Chang and Shoback, 2004).The activation of 

CaSR leads to cell signaling via different pathways; by coupling 

with G protein and activate Phospholipase C. Phospholipase 

C affects the phosphatidyl inositol 1, 4, 5-triphosphate (IP3), 

also mobilize Ca ions from the cell storage activation of protein 

kinase K and production of di-acylglycerol. Either acute or 

chronic exposure Cd ultimately modulates CaSR activities. Cd 

also inhibits Ca-ATPase activity. Na+, K+-ATPase is affected 

by Cd which ultimately leads to the general decrease in the 

cell membrane transport (Karthikeyan and Bavani, 2009). 

Cd shows higher affinity with sulfhydral group than 

phosphate, carbonyl, chloride or amino groups and as a result 

cadmium induces hepatotoxicity. By disrupting the 

intracellular redox state, toxicity can be produced by the 

activation of both protein and non-protein thiol group and as 

a result number of biological processes is affected. Redox 

balance shifts toward the oxidative state and oxidative stress 

is induced which cause number of deleterious effects. 

Oxidative stress is an imbalance state between the 

concentration of antioxidants defense mechanism and ROS 

concentration. Oxidative stress causes liver damage and Cd 

causes oxidative stress in liver. This also affects the prooxidantant 

and lipids per-oxidation balance by damaging the 

antioxidant barriers as shown in Fig 3. Non enzymatic antioxidants 

including total sulfhydral groups and GSH level is 

decreased by the Cd. As a result super dismutase, glutathione 

reductase, catalase and glutathione per-oxidase is inactivated. 

Amount of unbound free chelated Fe and Cu ions responsible 

for the promotion of oxidative stress also resulted by the Cd. 

By this way oxidative stress is induced in biological systems 

resulting in over production of ROS i.e.; hydroxyl radicals, 

hydrogen peroxide and superoxide ions. Very little knowledge 

is there about the exact mechanism and the evidences about 

free radical generation induced by Cd. Glutothionine is 

considered as the main thiol-disulphide redox buffer in the 

cells (Masella, et. al., 2005). 

GHS and heavy metal ion complex formation has entailed 

as the initiation of biological detoxification before the transfer 

to the cysteine rich peptides such as MT. the GHS capacity to 

regenerate antioxidant is linked to the redox state of GSSG/ 

2GSH. The decrease in hepatic GHS increases GSSG and 

decreased level of GHS/GSSG ratio increases Cd induced 

mortality and hepatotoxicity as shown in Fig 3. Cell death and 

GHS is lost in rat hepatocytes and in HepG2 cells on Cd 

exposure which indicate that GHS is the major element in the 

liver damage (Gebhardt, 2009). In normal conditions GSSG is 

regenerated to GHS by the help of flavor protein glutathione 

reductase which uses NADPH as electron donor. Cd inhibited 

GR activity is non competitive with respect to NADPH and 

GSSG and enhance Cd hepatotoxicity and mortality. 

Xenobiotics and their metabolites are detoxified by GSH which 

form compounds in conjugation with GHS either enzymatically 

or spontaneously with glutathione s-transferase (Mah and 

Jalilehvand 2010). Activity of antioxidant enzymes in the liver 

e.g.; SOD, CAT and GPx is also decreased with Cd. Possible 

mechanism which can explain increase in Fe is caused by Cd 

is the redistribution of Fe in the organism. Cd also binds with 

apoferritin and ferritin and displaces Fe from cellular sites (Lai 

Fig. 3. 

Cd decreases hepatic GSH which induce oxidative 

stress, which inhibit lipid per-oxidation as result 

hepatocellular damage is done. One the other hand Cd 

induced ROS block ETC in Mitochondria as a result 

less energy is produced and effects cellular functions.


and Loo, 2011). It has also been reported that stress genes 

and haem-oxygenase are unable to express fully by Fe after 

exposure to the Cadmium. 

There are several evidences that mitochondrion is the 

Key target on exposure to Cd (Belyaeva, et. al., 2011, Zhang, 

et. al., 2011). Mitochondria is chief source of Cd induced ROS 

production by blocking the electron transport chain in 

Mitochondria and by the acceleration of H+ influx through 

uncoupling oxidative stress. Mitochondrion is responsible 

for the production of energy. It interferes with the normal 

oxidative metabolism resulting in energy deficiency which 

affects various cell functions. Cd affects membrane’s structure 

function relationship by changing the lipids/Phospholipids 

profile and inhibits oxidative phosphorylation by changing 

function of Mitochondria (Modi and Katyare, 2009). Cd 

interacts with Mitochondrial membrane and produces 

conformational changes. As a result uptake of Ca uniporter– 

mediated cations is activated and Mitochondrial permeability 

transition is affected. The liver Mitochondria are considered 

as the major source of Cd induced ROS production (Belyaeva, 

et. al., 2006). 

Fig. 4. 

Cd interefers with Ca and inhibit Ca dependent AT 

Pases in endoplasmic reticulum. Ca level is increased 

which by dissipating Mitochondrial membrane potential 

and oxidative phosphorylation uncoupling can cause 

Apoptosis. In Mitochondria release of cyt c by Cd 

leads to apoptosis. ROS is released in response to Cd 

from Mitochondria and GSH level decreases. GSH 

induces oxidative stress which leads to Apoptosis. 

Cadmium Induced Apoptosis : 

Regulated form of cell death is called apoptosis. 

Apoptosis plays an important role in the maintenance and 

development of tissues and homeostasis in multicellular 

organisms. Cd induces apoptosis in liver which leads to 

hepatotoxicity (Lasfer, et. al., 2008) (Pham, et. al., 2006) (Yu, 

et. al., 2011). It is assumed that toxicity might be caused by 

the binding of thio-group to Cd in Mitochondria, which results 

Mitochondrial dysfunction leading to hepatotoxicity. Cd can 

also inhibit Mitochondrial functions i.e.; inhibition of 

respiration, MPT loss of trans-membrane potential, release of 

cyt-c. Cd may enter in Mitochondria through Ca uniporter 

and reacts with thiol group of adenine nucleotide translocator 

ANT which induces the MPT, cyt c release and apoptosis. 

Cyt c release and dysfunction of Mitochondria can suppress 

and delay in apoptosis (Oh and Lim, 2006). It is suggested 

that Mitochondrial mediated caspase independent pathway 

don’t inhibit apoptosis (Pham, et. al., 2006). In hepatoma cell 

line Hep-3 B Mitochondrial derived protein endonuclase-G 

(Endo G) is identified as potential caspase independent 

mediator in Cd induced apoptosis. This is done by the 

subsequent alteration in Ca and ROX of Mitochondrial 

homeostasis and release of Endo G and AIF (Lemarie, et al., 

2004). It is shown that Cd induced apoptosis is mediated by 

the release of Ca from the intracellular Ca storage but not an 

influx of extracellular Ca. Cd induces ER stress in vitro and in 

vivo and induces apoptosis (Kitamura and Hiramatsu, 2010). 

However, no exact data is present regarding the mechanism of 

Cd induced ER stress and its relationship with hepatocytes 

apoptosis (Biagioli, et al., 2008) as shown in Fig 4. 

Several studies have revealed that Cd is a toxic metal 

and caused imbalance in the redox state and as a result 

hepatocellular damage is done. Oxidative stress is due to 

imbalance in redox state. It is also thought that Cd is probable 

carcinogenic. Several questions are unaddressed regarding 

the exact mechanism of Cd induced damages. Cd competes 

with other metals in our body system and transported and 

disturbs the activities of or loss in antioxidant defense 

mechanism. Cd disturbs Ca homeostasis and leads to 

apoptosis. Cd also blocks ETC in mitochondria and effects 

the energy production. GHS production is depleted by Cd 

which leads to the aggravation of oxidative stress. Cd damages 

all liver cells. In order to set up proper therapeutic approaches 

we must know the proper mechanisms that how Cd cause 

chronic and acute toxicity. 

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Belyaeva, E.A., Dymkowska, D., Wieckowski, M.R., Wojtczak, L. 

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Received on 27.12.2012 Accepted on 15.02.2013



Role of Herbal Ice Cream in Human Health: A Review 

SWASHANKH KUMAR 1 , D C RAI AND DINKER SINGH 

Department of Animal Husbandry and Dairying, Institute of Agricultural Sciences, Banaras Hindu University, 

Varanasi 221 005, U.P., India. 

e-mail: shakyabhu68@gmail.com 1 

ABSTRACT 

Ice cream is a frozen dairy dessert that is made from products 

such as cream (or substituted ingredients) combined with 

flavourings and sweeteners. Herbal ice cream is gaining more 

popularity over synthetic products because of their functional, 

nutritional and pharmacological activities like gastrointestinal 

health, hypoglycaemic, immunomodulatory, anti-stress, 

analgesic, antipyretic, anti-inflammatory, antiulcerogenic, 

antihypertensive, radioprotective, antitumor and reinforcement 

of the immunological system. 

Key words: Herbal, Functional, Nutritional and Pharmacological 

Activities 

Ice cream is a frozen dessert that is delicious, nutritious 

and relatively cheap. The ice-cream industry in India is 

estimated to be worth 2000 cr, in which the branded market is 

100 million litres annually valued at 800 cr. The global market 

of ice-creams was pegged at US$61 billion in terms of retail 

value or 15 billion litres in terms of volume. The per capita 

consumption of ice-cream in India is just 300 ml/annum 

compared to 22 litres in the US or the world average of 2.3 

litres/annum (Soni, 2009). 

Ice Cream : 

Ice cream is a frozen dessert that is made from dairy 

products such as cream (or substituted ingredients) combined 

with flavourings and sweeteners such as sugar (Arbuckle, 

1986).Goff,et al., 1999, defined ice cream as a complex food 

colloid, containing fat globules, air bubbles and ice crystals 

dispersed in a freeze-concentrated dispersion D solution of 

proteins, salts, polysaccharides and sugars. 

History of Ice Cream : 

Ice cream had its origin in Europe and was introduced 

later in the United States where it is developed into industry. 

It is widely believed that ice cream evolved from iced beverage 

and water ices that were popular in the medieval period. It is 

know that wines and fruits juices were cooled with ice and 

snow brought from mountains to the court of the Roman 

Emperor Nero in the first century. Unfortunately, no definite 

description exits, except that snow and ice were used to cool, 

and possibly, to freeze sweet dessert. Over time, these water 

ices evolved into the popular frozen desserts of today. 

Swashankh Kumar is a Research 

Scholar at Department of Animal Husbandry 

and Dairying, Institute of Agricultural 

Sciences in Banaras Hindu University (BHU). 

He received M.Sc. Degree in Dairy 

Technology in 2007 from Institute of 

Agricultural Sciences, BHU, Varanasi, UP, India. His research 

interests include process optimization of herbal ice cream, 

development of a traditional product Malaeo. 

Preparation of Herbal Ice Cream : 

A detailed flowchart with mass balance and process 

details is given in Figure 1. 

Composition of Ice Cream : 

The composition of ice cream, which usually is 

expressed as a percentage of the constituents, varies 

depending on the market requirements and processing 

conditions.Approximate composition of commercial ice cream 

(%) given by Arbuckle, 1986 presented in Table 1. The 

Standards for microbiology analysis of ice cream is given in 

Table 2. 

Table 1. 

Approximate composition of commercial ice cream 

(%) given by Arbuckle, 1986 

Milk solid-not-fat/ 

Stabilizer & Total 

Milk fat 

Sugar 

Serum Solids 

Emulsifier Solids 

Economy Ice Cream 

10 10 to 11 13 to 15 0.30 to 0.50 35 to 37 

Good Average Ice Cream 

12 11 15 0.30 37.5 to 39 

Table 2. 

Standards for microbiology analysis of ice cream 

Parameter PFA rules 1955 

Total plate count < 2,50,.000 

Coliform count 

< 10/im 

E. coli Absent in 1 gm 

Salmonella 

Absent in 25 gm 

Stablylococcus aureus 

Absent in 25gm 

Yeast and mold count 


Anaerobic spore count 


Listeria monocytogens 

Absent in 1 gm

KUMAR et. al., : Role of Herbal Ice Cream in Human Health: A Review 131 

Health Benefits Provided by Ice Cream : 

Ice cream manufacturers and researchers have focused 

on producing highly palatable fat-reduced ice cream that meets 

the demand of health-conscious consumers (Prindiville,et al., 

2000).Ice cream fortified with fish protein (FP) can be a novel 

functional dessert with high nutritional value giving additional 

health benefits to a well-accepted traditional product. 

Davies, and Obafemi, 1985 investigated ice cream seems 

suitable for delivering probiotics in human diet because of its 

pleasant taste and attractive texture.Themechanism behind 

the specific health benefits of probiotic ice cream are related 

to gut micro flora modification, the strengthening the gut 

mucosal barrier, e.g. Adherence, pathogens inactivation, 

modification of dietary proteins by intestinal microflora 

modification of bacterial enzyme activity and influence on gut 

mucosal permeability and regulation of the immune system. 

Duthie,et al., 1981,reported ice cream might be the ideal dairy 

product to serve as a carrier for Lactobacillus acidophilus 

and is a better vehicle than milk. 

Now the trends to use of dietary fibre in ice cream 

increasing. Dietary fibre consists of nondigestible 

carbohydrates and lignin that are intrinsic and intact in plants. 

There are little data dealing with the study of the functionality 

of dietary fibres in ice creams (Thebaudin, et al., 1997).The 

physiological actions promoted by fibre addition in foods 

such as ice cream include the maintenance of gastrointestinal 

health, reduction of intestine transit time, protection against 

colon cancer, lowering of total and low-density lipoprotein 

cholesterol in the blood serum, reduction of postprandial blood 

glucose levels, increase of calcium Bioavailability and 

reinforcement of the immunological system (Tungland,and 

Meyer, 2002). 

The use of citrus fibre has led to significant improvement 

of melting quality of ice cream but failed to improve viscosity, 

overrun and texture (Dervisoglu,andYazici, 2006). In a similar 

study, the addition of rice flour was found to be a satisfactory 

fat replacer, though it imparted a powdery mouthfeel. However, 

there are limited data concerning the impact of dietary fibre on 

ice crystallisation and rubbery to glassy state transition 

phenomena. 

It is interesting to note that herbal ice cream contains 

similar health-promoting photochemical as do fruits and 

vegetables (Watson, 2001). Some studies were carried out to 

determine the minerals content of plants. Zengin,et al., 2008, 

concluded that aromatic plants are important sources of 

nutrients and essential elements. Some plants contained 

appreciable amounts of calcium, iron, zinc and potassium. 

Minerals such as Calcium is required for normal growth and 

skeleton development, aid in transmission of nerve impulses 

throughout the body; it is also required in association with 

phosphorus, sodium and potassium for water balance 

(Wardlaw, and Kessel, 2002). On the other hand, basil, fennel 

and yastimadhu contain appreciable amounts of minerals and 

trace elements. The role of minerals in our general health is 

now well demonstrated, on the other hand, the demand for 

herbs and spices is increasing now a days for their safety and 

culinary appeal (Vasudevan, and Sreekumari, 2007). 

Overrun : 

The overrun in ice-cream affects the body, texture and 

palatability of the final product. It is also related to the yield 

and profit. 

The addition of Herbal (Tulsi) extract intended to 

decrease the overrun significantly. All the samples could be 

statistically differentiated from each other. The lower overrun 

encountered in the experimental samples may be ascribed to 

the relatively higher viscosity associated with such samples 

(Das, et. al., 1989). 

Melting Resistance : 

Meltdown is also an important property of ice-cream 

affecting its sensory quality. It is important from at least two 

viewpoints (a) eye appeal and (b) mouth feel (Flack, 1988). 

Deviation in the melting property from ideal condition can 

make the ice-cream defective. Melting time was dependent on 

the ice-cream formulation and especially on the nature of the 

emulsifier. Fat aggregation appeared to be the major 

contributor to the melting resistance of ice-cream (Pelan, 

et al., 1997; Bolliger, et al., 2000; Goff, and Spagnuolo, 2001) 

through the existence of networks resulting from the presence 

of fat, proteins or other stabilizer. The melting of the ice was 

also controlled by the outside temperature and the rate of 

heat transfer. 

Melting resistance was obtained using the following 

equation: 

In general, as the viscosity increases, the resistance to 

melting and smoothness increases (Arbuckle, 1986). Slow 

melting generally indicates over stabilization and such 

condition can be corrected by reducing the amount of 

stabilizer and/or emulsifier. 

Structural Properties of Ice Cream : 

Ice cream is produced by freezing liquid ice cream mix. 

Ice cream has a complex physicochemical system (Koxholt,et 

al., 2001), containing fat globules, air bubbles and ice crystals 

dispersed in a freeze-concentrated dispersion of proteins, salts 

and polysaccharides (Goff, 1997). In particular, fat plays an 

important role in the stabilisation of ice cream structure by 

forming networks of fat globules surrounding the air bubbles 

(Koxholt,et al., 2001). The structure of the fats and their thermal 

characteristics is important factors in the behaviour of ice 

cream during freezing. In particular the destabilization of the 

fats is responsible for the creation of a network of fat globules, 

more or less coalesced, which contributes to the texture of the 

ice cream (Lucas,et al., 2005).


Air in ice cream provides a light texture and influences 

the physical properties of melt down and hardness and 

because of air incorporated in the ice cream mixture, the volume 

of the ice cream increases. Overrun is a measurement that 

shows increase in the volume of ice cream (Marshall, and 

Arbuckle, 1996). The percentage of overrun for ice cream is 

between 30 and 60% depending on total solids used in the 

formulation. Percentage of overrun increases with the amount 

of total solid (Marshall, and Arbuckle, 1996). 

Herbal ice cream has very high nutritive, characteristics 

flavor, taste, palatable nature and possible therapeutic value. 

Herbalice creamis very refreshing frozen dairy dessert 

particularly during summer months which improves the 

gastrointestinal health, hypoglycaemic, immuno-modulatory, 

anti-stress, analgesic, antipyretic, anti-inflammatory, 

antiulcerogenic, antihypertensive, radio-protective, antitumor 

and reinforcement of the immunological system. Keeping all 

the above nutritional benefit in mind we can say that herbal 

ice cream is the health food. 


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Publishing Company, Inc. 

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colloidal properties of ice-cream mix and ice-cream. Int. Dairy J., 

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Potato Pulp. Indian J. Dairy Sci., 42: 295-297. 

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London, U. K. Elsevier applied Science Publisher. pp. 83-107. 

Dervisoglu, M., and Yazici, F. 2006.The effect of citrus fibre on the 

physical, chemical and sensory properties of ice cream. Food Science 

and Technology International, 12: 159–164. 

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An ideal vehicle for lactobacillus acidophilus. Dairy Field, 164: 

139-140. 

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fat globule sizes, on meltdown of ice cream. J. Dairy Sci., 84: 31–37. 

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York: International Thompson Publishing. 

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stability of aerated milk protein emulsions in the presence of small 

molecule surfactants. J. Dairy Sci., 80: 2631–2638. 

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Publications Pvt. Ltd. 

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milk fat, cocoa butter, and why protein fat replacers on the sensory 

properties of low fat and nonfat chocolate ice cream. J. Dairy Sci., 

83: 2216-2223. 

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53. 

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1997. Dietary fibres: Nutritional and technological interest. Trends 

in Food Science and Technology, 8: 41–49. 

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polysaccharides (dietary fibre): Their physiology and role in human 

health and food: Comprehensive Reviews.Food Science and Food 

Safety, 1: 73–92. 

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LTD., New Delhi. pp. 316-317. 

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McGrow Hill. 

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CRC Press LLC. 73: 180. 

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of some aromatic plants, their growth soils and infusions. J. Sci. 

Food and Agriculture, 88(4): 581-589. 

Received on 17.01.2013 Accepted on 12.03.2013



Management of Stored-product Insect Pests Through Biorational Approaches: 

A Review 

S. MURALI 

Department of Entomology, UAS, GKVK, Bangalore 560 065, Karnataka, India 

e-mail: dr.mmrl@rediffmail.com 

ABSTRACT 

Stored-product insects are serious pests of dried, stored, durable 

agricultural commodities and of many value-added food 

products and non-food derivatives of agricultural products 

worldwide. The effect of diatomaceous earth on mortality of 

adults of Sitophilus oryzae clearly revealed the importance of 

temperature and relative humidity and found hundred per cent 

mortality even at highest RH (90%) at both highest temperatures 

(30 and 35ºC) and at combinations of 20 and 25ºC with only 

lower RH (30 and 50%). The exposed adults of Sitophilus oryzae 

and Rhyzopertha dominica and adults and eggs of Corcyra 

cephalonica to essential oils of geranium, lemongrass and 

peppermint in the fumigation chamber noticed that complete 

mortality of adults of S. oryzae at 100 and 150 ìl/250 ml of 

peppermint oil. Whereas, in case of R. dominica, 100% mortality 

was observed in all the doses (50, 100, 150 & 200 ìl/250 ml). The 

effectiveness of controlled atmosphere (decreased oxygen at 

1± 0.5%) at elevated temperature (41ºC) in controlling the 

major storage pests, fig moth (Ephestia cautella), Indian meal 

moth (Plodia interpunctella), and dried fruit beetle (Carpophilus 

sp.) showed effective control. 

Key words Stored-product insects, biorational pest management, 

Temperature, Relative humidity, Controlled atmosphere. 

Murali, S. presently pursueing Ph.D. 

degree in the Department of Agricultural 

Entomology at UAS, GKVK, Bangalore. He has 

been completed masters degree at Bangalore, 

GKVK. During M. Sc. my research on 

prospecting antimicrobial peptides from aculeate 

Hymenoptera: In vivo screening of antimicrobial peptides 

from bees and wasps. Now presently working on 

management of Brinjal shoot ant fruit borer at NBAII, 

Bangalore. 

insects causes to dried-stored durable agricultural 

commodities, Value-added food products and Non-food 

derivatives of agricultural products. 

Economic loss in Stored grains: 

There is estimation of annual loss -14 million tonnes. 

i.e., Rs. 7,000 crores and Insects alone causes Rs.1, 300 crores 

and estimated to cause 600 species of beetle pests and 70 

species of moths & about 355 species of mites. 

External Feeders: 

Stored-product insects are serious pests of dried, stored, 

durable agricultural commodities and of many value-added 

food products and non-food derivatives of agricultural 

products worldwide. Stored-product insects can cause serious 

postharvest losses, estimated from up to 9 per cent in 

developed countries to 20 per cent or more in developing 

countries, but they also contribute to contamination of food 

products through the presence of live insects, insect products 

such as chemical excretions or silk, dead insects and insect 

body fragments, general infestation of buildings and other 

storage structures and accumulation of chemical insecticide 

residues in food, as well as human exposure to dangerous 

chemicals as a result of pest control efforts against them 

(Rajendran, 2002). 

Khapra beetle 

(Trogoderma 

granarium) 

Internal Feeders: 

Saw toothed grain 

beetle 

(Oryzaephilus 

surinamensis) 

Red flour beetle 

(Tribolium 

castaneum) 

Stored product Insects: 

They are categorised as Internal feeders, External 

feeders, Scavengers and Secondary pests. Stored product 

Rice weevil 

(Sitophilus oryzae) 

Lesser grain borer 

(Rhizopertha 

dominica) 

Pulse beetle 

(Callosobruchus 

chinensis)


High Temperature: Temperature above 45 C - insects 

die within 24 h, Monitoring of the temperatures is often done 

by handheld electronic thermometer and super heating system 

by infrared heaters in storage godowns at 55-60 0 C for 10 to 20 

minutes. 

Response To Stored Product Insects To Temperature: (Fields, 

1992) 

Zone Temperature Effect 

Lethal 50-60 Death in minutes 

45-50 Death in hours 

Sub optimal 35 Development stops 

33-35 Development slow 

optimal 25-33 Max Rate of devt 

Development stops 13-25 Development slow 

13-20 Development stops 

Lethal 5 Death in week 

-10- -5 Death in days 

-25 - -15 Death in minutes 

Biorational Approaches: 

Safe, Effective and Relatively simple. 

Monitoring: 

Mainly it helps to avoid unnecessary pest management 

expenses, Determining when and where insect pest 

suppression will be needed. Sampling – Best tool to provide 

this information and no. of insects in samples or % of samples 

infested is used to estimate overall level of insect infestation. 

Sanitation and Exclusion: 

Sanitation of grain and food storage facilities and the 

effective exclusion of stored-product insects from such 

structures and from food packages are the keys to preventive 

management of storage insects. For bulk-stored grain it is 

imperative that newly harvested commodities be stored in 

clean bins and not be loaded into bins that contain older 

products that may harbour insects. 

Temperature Management: 

Insect populations can be managed by manipulating 

the temperature of their environment. The maximum rate of 

growth and reproduction for most insects occurs between 25 

and 33æ%C and is reduced at temperatures above and below 

this range, with complete cessation of development and 

eventual death at approximately 13 and 35æ%C. Maximum 

rate of growth & reproduction for insects 25 and 33 °C, 

Cessation of development & eventual death 13 and 35 °C and 

two ways of temperature - Low Temperature and High 

Temperature. 

Low Temperature: Mainly through turning grain over, 

ambient air aeration or chilled aeration and Force cool air 

through the grain bulk. 

Irradiation: 

Irradiation of durable stored products is legal in most 

countries and can be conducted using ionizing radiation such 

as gamma rays, which have the potential to dislodge electrons 

from chemical bonds in molecules, and nonionizing radiation 

such as radio frequencies, microwaves, or infrared rays, which 

do not break bonds but essentially heat the product and the 

insects by vibrating bonds in water. Irradiation could be used 

to disinfest product entering a grain storage system or as a 

remedial treatment for infested product in a storage system. 

Irradiation technqnics used storage pest control mainly 

includes Infra red, Ultra violet lights (

MURALI : Management of stored-product insect pests through biorational approaches: A review 135 

Advantages: 

Mainly includes no residues in treated products, workers 

safety issues, environmentally safe and Low risk for resistance 

development. 

Disadvantages: 

Mainly includes high gas tightness required, longer 

treatment times, increased production (GAS) costs and 

logistical problems related to material availability and supply. 

Insect Growth Regulators (IGRS): 

Insect growth regulators (IGRs) used in stored product 

systems in the United States and elsewhere include the insect 

juvenile hormone analogs methoprene, hydroprene, and 

pyriproxyfen. All three compounds mimic the effects of 

sustained increased titer of insect juvenile hormone by 

disrupting normal development between larval instars and in 

metamorphosis from larvae to pupae and then from pupae to 

normal adults. These IGRs are not directly toxic to adults, 

although their potential effects on reproductive sterility have 

not been fully investigated. Another key attribute to these 

IGRs is their low levels of toxicity to mammals and inherent 

high level of food safety. 

Insect juvenile hormone analogs methoprene, 

hydroprene and pyriproxyfen and they act by disrupting 

normal development – Between larval instars, metamorphosis 

from larvae to pupae and from pupae to normal adult. 

Application Of IGR’S: 

Hydroprene (Gentrol) - effective against late-instars of 

Plodia interpunctella, Tribolium castaneum and T. confusum. 

As surface treatments to flooring structures of mills, 

warehouses and processing plants for example Methoprene 

and Pyriproxyfen and Used as aerosol spray treatments. 

Biological Control: It includes mainly Pathogens, 

Parasitoids and Predators. 

Microbial Insecticides: 

Natural occurrence of entomopathogenic fungi and 

bacteria has been reported among storage pests. Commercially 

available fungi Beauveria bassiana, Metarhizium anisopliae 

and Bacillus thuringiensis alone or in conjunction with others 

can control some stages of storage pest species. 

Spinosad: 

Spinosad is an insecticide derived from metabolites in 

the fermentation of the actinomycete bacterium, 

Saccharopolyspora spinosa Mertz and Yao (Actinomycetales: 

Actinomycetaceae). Spinosad is currently registered by the 

U.S. There is much interest in the use of spinosad on stored 

grain because other residual insecticides registered in the 

United States and elsewhere have limited efficacy against the 

major pest of stored wheat, R. dominica, either because of 

simple lack of efficacy or because of development of 

resistance. Spinosad is effective for season long control of R. 

dominica in stored wheat; it is highly toxic to larvae of many 

stored-product insects and shows good compatibility with 

insect natural enemies. It is a bacterial fermentation product - 

Saccharopolyspora spinosa and it acts by Ingestion and 

contact poison; Effective for season long control of R. 

dominica in stored wheat. 

Important Parasitoids: 

Several species of parasitoid wasps from the 

Pteromalidae are solitary ectoparasitoids of internal-feeding 

grain-infesting species of beetles, and similarly there are 

several common species of Ichneumonidae and Braconidae 

as ecto- and endoparasitoids associated with stored-product 

Lepidoptera. Some species of free-living predatory beetles, 

true bugs (Heteroptera: Anthocoridae), and mites prey on any 

life stage of numerous species of stored-product insect pests 

that they can subdue and consume. Populations of parasitoids 

and predators in storage systems display delayed density 

dependency in their dynamics that are typical of other predatorprey 

and parasitoid-host systems in other insect communities, 

and population declines of stored product pest species are 

typically followed by increase in these natural enemy 

population. It includes Braconidae, Ichneumonidae, 

Pteromalidae and Bethylidae. The Parasitic wasp, 

Anisopteromalus calandrae - reduced rice weevil infestation 

of spilled wheat by 90% in a simulated warehouse. Almond 

moth, Cadra cautella populations were supressed to extend 

of 97.3% of due to B. hebetor release and Egg parasitoid, 

Trichogramma pretiosum - against Cadra cautella, Ephestia 

spp. and P. interpunctella and T. pretiosum suppressed 

C. cautella upto 42% and P. interpunctella upto 57%. 

Botanicals: 

Farmers often use homegrown or naturally occurring 

plant materials for insect control in developing countries. 

Problems with botanical insecticides are lack of consistency, 

safety concerns, and sometimes odor. It is often falsely 

assumed that because a plant material is used as a food 

flavoring or medicine that extracts from the material will be 

safe for human consumption. Various extracts from the neem 

tree, Azardirachta indica, collectively referred to as the 

insecticide Neem, are commercially available botanical 

insecticides, and local formulations have been widely used in 

some parts of the world for stored-product insect control. 

Plant derived compounds acts as ovicidal, repellent, 

antifeedant, sterilization and toxic effects in insects. 

Plant extracts: 

Root powder of sweet flag, Acorus calamus L., with 

activity against stored grain beetles and leaves of Azadirachta 

indica with protectant properties against stored grain beetles 

and Angoumois grain moth, Sitotroga cerealella in wheat.


Varietal Resistance: 

Hull integrity is the best predictor of rice resistance to 

R. domnica and Phenolic content (corn) related to kernel 

hardness - resistance to the maize weevil, S. zeamais and the 

larger grain borer, Prostephanus truncates. 

Edible Oils: 

Obtained from leaves or seeds of plants are often applied 

to stored products and volatility and insecticidal efficiency of 

essential oils – act as “fumigant”. 

Inert Dusts (ID’S): 

IDs are powders that render the insects more sensitive 

to desiccation and divided into four group’s viz., Clays and 

ashes, Minerals, Diatomaceous earth (DE) and Synthetic silica 

aero gels. 

Modes of action: 

Mainly acts by Asphyxiation through blocking of 

spiracles leads to the insects killing, Abrasion of cuticle leads 

to increase water loss and dusts absorb water and epicuticular 

lipids of arthropods leading to excessive water loss through 

the cuticle. 

Diatomaceous Earth (DE): 

Many tests have been conducted to synergize 

pathogens with other control technologies, particularly those 

that might be expected to increase efficacy of pathogens, such 

as DE by presumably abrading the cuticle, or grain varietal 

resistance by delaying larval development, both of which 

might make the insect more susceptible to the pathogen. 

Fossilized remains of the silicon dioxide skeletons of 

diatoms, which are aquatic algae that are insecticidal as 

desiccants. Mode of action through killing of insects by 

absorbing hydrocarbons from their cuticles causes 

dehydration and death; has very low mammalian toxicity and 

add 3-4 Kg. of Diatomaceous Earth to each tonne of grain for 

effective control of pests in storage. 

Activated Clay: 

Raw clay (Kaolinate clay) is used in the ratio of 45% 

silicon dioxide + 38% aluminium oxide for effective control of 

pests; Grain purpose use activated kaolin 1 kg/100 kg of grain 

and Seed purpose mix 1 Kg of Activated Kaolin with every 

100 Kg of seed and pack it. 

Pheromones: 

Attractant pheromones, which are intraspecific chemical 

signals, and other attractant semiochemicals have been 

identified for over 40 species of stored-product insects over 

the past 40 years. There are two broad categories of 

pheromone systems recognized in stored-product insects, 

which follow life-history models for insects in general. Species 

with short-lived, usually nonfeeding adult stages utilize femaleproduced 

sex pheromones in which a receptive adult female 

“calls” by releasing one or more attractant compounds and 

one or more males respond upwind to the pheromone after 

which mating occurs. 

Categories: 

1. Species with short-lived (< 1 month) 

2. Species with long-lived (> 1 month) 

Short-lived: 

Non-feeding adult stages utilize female-produced sex 

pheromones to attract males 

Eg: Pyralidae, Anobiidae, Bruchidae and Dermestidae 

Long-lived: 

Feeding adults utilize male produced aggregation 

pheromones to attract both males and females 

Eg:Bostrichidae, Curculionidae, Cucujidae, Sylvanidae 

and Tenebrionidae. 

Insects 

Trogoderma sp. 

Rhizopertha dominica, Protsephanus truncatus 

Sitophilus oryzae 

Cadra cautella, Plodia interpunctella 

Lasioderma serricorne 

Pheromones Based On Chemical Groups: 

Traps: 

Lures 

Trogodermal 

Dominicalure 

Sitophilure 

TDA 

Serricornin 

Traps are designed to sit on a floor or flat surface and 

capture insects that walk into the trap, which eventually 

become stuck to the trapping surface or ensnared inside the 

trapping receptacle. 

Barak and Burkholder developed a trap with horizontal 

layers of corrugated cardboard in which responding beetles 

walked through the tunnels of corrugations to reach a cup of 

oil into which they fell and became suffocated. Traps can be 

used to monitor or directly reduce insect populations and 

typically use food, visual lures, chemical attractants and 

pheromones as bait. 

Impact and Removal: 

Internal and external feeding insects killed by impact 

and Entoleters & pneumatic conveyers used to move grains 

from one bin to another. Mechanism of impact through 

commodities are fed into the center of rotor of an impact machine 

this can be accelerated by centrifugal force thus insects killed 

when they hit the pegs on rotor and removed by aspiration or 

sieving. 

Thomas and James 2010 reported that stored-product 

insects are ubiquitous, essentially cosmopolitan, occurring 

in feral habitats as well as in human-made facilities and

MURALI : Management of stored-product insect pests through biorational approaches: A review 137 

infestation can be a continual year-round process that makes 

pest control difficult. Bio-based pest management methods is 

available for stored-product systems, including inert 

diatomaceous earth (DE) as an insecticidal desiccant, the 

microbial insecticide spinosad, highly safe synthetic insect 

growth regulators (IGRs), controlled and modified 

atmospheres as alternatives to traditional chemical fumigants, 

insect natural enemies that can regulate or control pest 

populations, pheromones and other semio-chemicals that can 

be used in traps for monitoring or applied as control tactics in 

mating disruption or attract-and-kill. 

Studies of Poornima and Avaknavar 2008 on the effect 

of DE on mortality of adults of Sitophilus oryzae clearly 

revealed the importance of temperature and relative humidity. 

They found hundred per cent mortality even at highest RH 

(90%) at both highest temperatures (30 and 35ºC) and at 

combinations of 20 and 25ºC with only lower RH (30 and 50%). 

Michaelraj, et al. 2008 in their studies exposed the adults of 

Sitophilus oryzae and Rhyzopertha dominica and adults and 

eggs of Corcyra cephalonica to essential oils of geranium, 

lemongrass and peppermint in the fumigation chamber. They 

noticed complete mortality of adults of S. oryzae at 100 and 

150 ìl/250 ml of peppermint oil. Whereas, in case of R. dominica, 

100% mortality was observed in all the doses (50, 100, 150 & 

200 ìl/250 ml). 

Fatih, et al., 2010 reported the effectiveness of controlled 

atmosphere (CA) (decreased oxygen at 1± 0.5%) at elevated 

temperature (41º C) in controlling the major storage pests, fig 

moth (Ephestia cautella), Indian meal moth (Plodia 

interpunctella), and dried fruit beetle (Carpophilus sp.). The 

CA treatment can be recommended as a post-harvest methyl 

bromide (MB) alternative for dried figs since it provided 100% 

control of the pest species tested, had neutral or positive 

effects on dried fruit quality and required comparatively short 

treatment times compared with other MB alternatives. 


Fatih Sen., Kamer, B., Ferit Turanli. and Yugun Aksoy., 2010, Effects 

of short term controlled treatment at elevated temperature on 

dried fig fruit. Journal of Stored Products Research., 46: 28-33. 

Grenier, A. M., Pintareau, B. and Nardo, P., 1994, Enzymatic variability 

in three species of Sitophilus oryzae (Coleoptera: Curculionidae). 

J. Stor. Prod. Res., 6: 201-213. 

Michaelraj, S., Kirti Sharma. and Sharma, R. K., 2008, Fumigant toxicity 

of essential oils against key pests of stored maize. Ann. Pl. Protec. 

Sci., 16: 356-359. 

Poornima, V. and Avaknavar, J. S., 2008, Effect of temperature and 

relative humidity on efficacy of diatomaceous earth on mortality 

of rice weevil, Sitophilus oryzae. Karnataka J. Agric. Sci., 22: 99-103. 

Thomas, W. and James, E., 2010, Biorational approaches to manage 

stored-product insects. Annu. Rev. Entomol., 55: 375-397. 



Preliminary Studies of Phenology of Some Selected Tree Species from Ngel-Nyaki 

Forest Reserve 

MOHAMMED, S., ALIYU, B., UMAR, M.D. 

*Department of Biological Sciences, Gombe State University, PMB 0127. Nigeria. 

e-mail: sumulsu@yahoo.com, aliyubabale@yahoo.com and danladi.umar@pg.canterbury.ac.nz 

ABSTRACT 

A checklist of some selected tree species as data bank on plant phenology (i.e. leafing, flowering and fruiting) at Ngel-Nyaki forest 

reserve was carried out. Sixteen individual trees belonging to eight species of angiosperm were selected for the study through six 

months duration. Variation in flowering related to leaf flushing revealed three flowering types; Nov-Dec, Dec-Mar and Jan-Mar. A 

wide range of producton of new leaves were seen in some species and reproductive phase were correlated to see the relation. 

Available phenological information on the sixteen individual trees showed that Nov-Dec flowering species were most abundant 

among the three types recognize. Among species are; Anthocleista vogellii, Bridelia speciosa, Trema orientalis and Dombeya 

ledermannii. Period of production of new leaves, flowering and fruiting were positively correlated in seven species. Correlation of 

same in Anthonotha noldeae was insignificant (r=-0.0095). 

Key words Preliminary, Phenology, Ngel-Nyaki, montane, Forest reserve 

Phenology is an area of science that has receive a 

renewed interest in recent years. It is defined as the study of 

cyclic and seasonal natural phenomena, especially in relation 

to climatic and animal life (Primack, 1987). The return of 

migratory songbirds, the blooming of wild flower and woody 

landscape plants and the development of locally indigenous 

insect are all examples of phenological events which are easily 

observed each spring in any location. 

Changes in timing of such events; leafing, flowering 

and fruiting can indicate climatic change. For instance,the 

data on Anthonotha noldeae, showed flowering on-set every 

Nov through Feb. But in 2008, June at Ngel-Nyaki main forest 

three individual trees have been spotted with flowers, while a 

few with flower buds (NMFP Station,2008). Climate change 

affects individual plants species by changing the speed and 

duration of physiological (life) processes such as growth rate, 

the degree of evaporation of available water and their 

interaction with birds and other insects. Phenology has 

recently become very popular in climatic change studies worldwide 

(Chapman, et al., 2004). 

Many of these are endemic to Afromontane region, four 

tree species, including Anthonotha noldeae were new to west 

Africa and other new to Nigeria. Diversity is reflected in the 

high number of primates and other animal diversity (Chapman 

and Chapman, 2001). 

Complete phenology study must be all year round and 

this research was carried out for a period of six months as a 

preliminary. As the data are scanty with respect to checklist of 

Nigerian montane forest, so the aim of this work is to establish 

a checklist of some selected tree species as data bank on 

phenology (onset of leaf development, flowering and fruiting 

duration) at Ngel-Nyaki forest so as to facilitate an 

understanding of their growth cycle within the forest and 

surrounding fragments. 

MATERIALS AND METHODS 

Study Area 

The study was conducted at Ngel-Nyaki forest reserve 

and it’s surrounding riverside fragments. It is the most diverse 

forest on Mambilla Plateau. Ngel-Nyaki forest is located 

between Longitude 11 0 00’ and 11 0 30’ East and Latitude 6 0 30’ 

and 7 0 15’ North and has 1,400-1,500m elevation on the Western 

escarpment of Mambilla Plateau. It located in the South-East 

corner of Taraba State Nigeria and mesures about 3100km 2 . 

The reserve experiences an avearge rainfall of 250days, 

between the months of March and October. Mean annual 

rainfall of 1,780mm peaks between the the months of June and 

July. Temperature has never exceeded 30 0 c. Over 146 vascular 

plant species were collected, many of which are endemic to 

the Afromontane region (Chapman and Chapman, 2001). 

The reserve lies on the West facing slopes of an old 

volcano, between 1,650m to 1,450m elevation and it is 6.6km 2 in 

area (actual forest), while Ngel-Nyaki forest reserve is 46km 2 . 

Apart from plant diversity which is reflected in the high number 

of primates and other animal species (Beck and Chapman, 

2008). Ngel-Nyaki forest is also home to a good number of 

birds (e.g. collared songbird, green headed songbird, northerndouble 

collared songbird,turacos, barbets, bulbuls etc.) and 

wintering site to some paleactic and neactic migrants 

(Chapman and Chapman,2001, Hamilton, 1975 and Richards, 

1957). 

Experimental design and selected tree species with their 

respective sites : 

A total of 16 individual trees of 8-species (i.e. duplicate) 

belonging to the seven-families of Angiosperm were 

phenologically censured. The plants were selected randomly 

in both main forest and two surrounding fragments (i.e.

MOHAMMED et. al., : Preliminary studies of phenology of some selected tree species from Ngel-Nyaki forest reserve 139 

fragment B and C). Method of data collection differs based on 

research interest (Newstrom, et al., 1994 and Schwartz, 2003). 

The plants are selected randomly in both the main forest 

and two surrounding fragments and tagged for observation 

on leafing, flowering and fruiting. Each tree species is 

replicated twice (i.e. two stand per species). 

The location of the plant species are identified as follows: 

1, tree species in the fragment A and B; 

Anthocleista vogellii 

Croton macrostachyus 

Dombeya ledermannii 

Syzygium guineence 

Trema orientalis 

2, tree species occur in both main forest and fragments; 

Albizia gummifera 

Anthonotha noldeae 

Bridelia speciosa 

Data collection/Record of reproductive events : 

A scale of 1-4 was employed insuring at two weeks 

interval for record of events i.e. Leafing, flowering and fruiting, 

where 1 is 25% and 4 is 100% of the events. A pair of Binocular 

was used for viewing and a record sheet. Each tree was scored 

as new/no leaf, flower bud, flower, immmature fruit and mature 

fruit. Correlation Co-efficient statistical analysis was used 

between two individual thesame species (i.e. r=) 

RESULTS AND DISCUSSION 

Discription of the Result, best on the two individual 

trees of thesame species and their phenological relation. 


Fragment C tree: in Nov.; 37% no leaf and 62.5% new 

leaf, Dec.; 62.5% new leaf, Jan; 25% no leaf, Feb.; 12.5% no 

leaf, Mar.; 12.5 no and new leaf and Apr.; 25% new leaf. 

Main forest tree: Nov.; 75% new leaf and 25% flower 

bud, Dec.; 50% no leaf and 37.5% new leaf, Jan.; 25% new 

leaf, 37.5% flower bud and 12.5% flower, Feb.; 50% flower, 

Mar.; 37.5% flower bud, 25% flower and 25% immature fruit, 

Apr.; 12.5% immature and mature fruit. 

The species in fragment C and main forest; r= 0.5799 


Fragment B tree: Nov.; 12.5% no leaf and flower bud, 

37.5% immature and mature fruit, Dec.; 25% flower bud, 37.5% 

flower, 25% immature fruit and 12.5% mature fruit, Jan. 37.5% 

flower bud, 12.5% flower and 25% immature and mature fruit, 

Feb.; 25% flower, 50% immaature and 25% mature fruit, also 

the same in March and April. 

Fragment C tree: Nov.; 12.5% new leaf and flower bud 

and 37.5% immature and mature fruit, Dec.; 12.5% new leaf 

and 50% mature fruit, Jan.; 12.5% new leaf and flower and 

50% mature fruit, Feb.; 12.5% flower and immature fruit and 

50% mature fruit, Mar.; 25% flower, immature and mature fruit, 

April; 25% flower, 50% immature fruit and 25% mature fruit. 

The species in fragment B and C, r=0.6725 


Fragment C tree: 25% flower bud and flower in Nov., 

Dec., Jan., Feb. and Mar. The individual shed its leaf about 

12.5% and resting period in April. 

Main forest tree: Nov.; 37.5% flower bud and 50% flower, 

Dec.; 62.5% flower and 25% immature fruit, Jan.; 25% no leaf 

and 50% immature fruit, Feb.; 25% no leaf, 12.5% new leaf and 

50% immature fruit, Mar.; 12.5% no leaf and new leaf and 50% 

immature fruit, Apr.; 25% no leaf and new leaf and 50% immature 

fruit. 

The species in fragment C and main forest; r= - 0.0095 


Fragment C tree: Nov.; 12.5% new leaf, 50% flower bud 

and 37.5% flower, Dec.; 12.5% flower bud, 50% flower and 

12.5% immature fruit, Jan. and Feb.; 12.5% no leaf and immature 

fruit, Mar.; 12.5% no leaf and 50% immature fruit, Apr.; 50% 

immature fruit. 

Main forest tree: Nov.; 50% flowerbud and flower, Dec.; 

37.5% flower bud, 25% flower and immature fruit, Jan. 25% 

immature fruit, Feb.; 50% immature fruit, Mar.; 12.5% immature 

fruit, Apr.; 62.5% new leaf. 

The species in fragment C and main forest, r= 0.9197 


Fragment B tree: Nov. and Dec.; 25% new leaf, Jan.; 

12.5% no leaf and new leaf, Feb.; 25%no leaf and 12.5% new 

leaf, Mar.; 25% no leaf and Apr.; 50% no leaf. 

Fragment C tree: Nov.; 37.5% no leaf and 25% new leaf, 

Dec.; 50% new leaf, Jan.; 12.5% no leaf and 37.5% new leaf, 

Feb.; 37.5% no leaf and 12.5% new leaf, Mar. and Apr.; 12.5% 

new leaf and 50% no leaf. 



Fragment B tree: Nov.; 37.5% new leaf, 25% flower bud 

and 12.5% flower, Dec.; 37.5% flower bud, 50% flower and 

12.5% immature fruit, Jan.; 62.5% immature fruit and 37.5% 

mature fruit, Feb.; 75% mature fruit, Mar.; 25% no leaf and 

25% mature fruit and 50% no leaf in April.


Fragment C tree: all the reproductive events are thesame 

as the above fragment B tree. 

So, r = 1 

Syzygium guineense 

Fragment B tree: Nov.; 37.5% no leaf and 12.5% new 

leaf, Dec.; 25% no leaf and 75% new leaf, Jan.; 50% new leaf 

and 25% flower bud, Feb.; 50% flower bud and 37.5% flower, 

Mar.; 37.5% flower bud and flower and 25%immature fruit, 

Apr.; 25% flower and 50% immature fruit. 

Fragment C tree: Nov.; 37.5% no leaf and 12.5% new 

leaf, Dec.; 25% no leaf and 62.5% new leaf, Jan. and Feb.; 25% 

new leaf and 50% flower bud, Mar.; 37.5% flower bud, 50% 

flower and 12.5% immature fruit, Apr.; 25% flower and 50% 

immature fruit. 



Fragment B tree: Nov.; 12.5% flower bud and flower, 

50% immature fruit and 12.5% mature fruit, Dec.; 12.5% flower, 

62.5% immature fruit and 25% mature fruit, Jan.; 12.5% flower 

bud, 25% flower and 62.5% immature fruit, Feb.; 12.5% flower 

bud, 37.5% flower and 50% immature fruit, Mar.; 12.5% flower, 

62.5% immature fruit and 25% mature fruit, Apr.; 37.5% 

immature fruit and 62.5% mature fruit. 

Fragment C tree: Nov.; 12.5% flower bud, 37.5% flower, 

37.5% immature fruit and 12.5% mature fruit, Dec.; 50% immature 

and mature fruit, Jan., Feb. and Mar.; 12.5% flower, 62.5% 

immature fruit and 25% mature fruit, Apr.; 37.5% immature 

fruit and 62.5% mature fruit. 

The specie sin fragment B and C, r=0.8832 




Fig. 1, reproductive events among individual trees in 

main forest of Ngel-Nyaki forest between the months of Nov.- 

April. 

Keys 


Croton macrostahyus 




Fig. 2, reproductive events among individual trees in 

fragment B of Ngel-Nyaki forest between the months of Nov.- 

April. 

Keys 





Fig. 3i. 

Keys 





Fig. 3ii. 

Fig. 3 (i and ii), reproductive events among individual 

trees in fragment C of Ngel-Nyaki forest between the months 

of Nov.-April. 

The occurence of leaf/leaf flushing (vegetative phase) 

and flowering (reproductive phase) requires the availability 

of substantial amount of resources within the trees. Flower 

production and maintenance requires considerable expense 

of energy to form non-photosynthetic tissue and nectar 

(Knight, et al., 2005). 

The result of this research work, agrees with the findings 

of (Hamza, 1990 and Keay, 1989) which indicates that Albizia 

gummifera, Anthocleista vogellii and Croton macrostachyus 

leaf in the months of Nov. and Feb. The duration of leaf 

shedding (no leaf) and time lag between the onset of leafing 

and flowering help in making use of available resources for 

growth and reproduction. 

Flowering and fruiting in fragment B, C and main forest 

in tree species typically occur throughout the six months, 

may be on one species or another. Synchronization of 

flowering with a particular season of the annual cycle by 

Anthocleista vogellii and Trema orientalis appear to be under 

the control of prevailing climatic conditions of the season. 

The detection of the several flowering types in fragment B, C 

and main forest trees revealed that Bridelia speciosa, Croton 

macrostachyus, Syzygium guineense and Trema orientalis 

flowered at the end of rainy season. This result agrees with 

the findings of Hamza 1990 and Keay 1989. The proportion of 

species flowering during dry period of the year varies widely 

among tropical deciduous forest trees bearing different 

intensity of drought. As the case of Albizia gummifera, 

Bridelia speciosa, Dombeya ledermannii and Trema 

orientalis. 

Fruit maturation and suitable condition(s) for dispersal 

are closely synchronized in tropical dry forest because of the 

pronounced differences of abiotic and biotic conditions 

between dry and rainy seasons. As Trema orientalis and 

Anthonotha noldeae flower in Nov, produce fruit in Dec.- 

April. This result agrees with the findings of Chapman and 

Chapman 2001 and Lieberman and Lieberman, 1987 who

MOHAMMED et. al., : Preliminary studies of phenology of some selected tree species from Ngel-Nyaki forest reserve 141 

reported that different flowering are types related to varying 

duration of fruiting phenophase. Thus; all flowering types 

complete the fruiting phenophase before the onset of the 

succeeding rainy season, ensuring that some (if not all) seeds 

are available for germination when the soil is sufficiently moist. 

Phenological events in tropical trees may defend on 

preceeding and successive stages; hence the fruiting duration 

may impose constraints on flowering time. Although, fruiting 

duration is related to flowering time, especially in species 

flowering in dry season of the year. As the case of Albizia 

gummifera and Syzygium guineense (Hamza, 1990). 

The variety of seasonal flowering of the tree species at 

Ngel-Nyaki forest with linkages to leaf flush time and leafless 

period reflect the fact that variable reproductive and survival 

strategies evolved in tree species under climatic condition. 

Determine the flowering and fruting period of the tree species 

will help to devise appropriate strategies for conserving the 

plants at Ngel-Nyaki forest which are of positive value. 


Beck, J. and Chapman, H. 2008. A population estimate of the endangered 

chimpanzee Pan troglodytes vellerosus in a Nigerian montane forest: 

implications for conservation. Oryx, 42: 448-451. 

Chapman, H. M., Olson, S. M. and Trumm, D. 2004. An assessment of 

changes in the montane forests of Taraba State, Nigeria, over the 

past 30 years. Oryx, 38, 282-290. 

Chapman, J. D. and Chapman, H. 2001. The forests of Taraba and 

Adamawa States, Nigeria: an ecological account and plant species 

checklist, Dept. of Plant and Microbial Sciences, University of 

Canterbury. 

Hamilton, A. C. 1975. The significance of patterns of distribution 

shown by forest plants and animals in tropical Africa for the 

reconstruction of palaeoenvironments: a review, AC Hamilton. 

Hamza, M. 1990. Trees and shrubs of the Sudan. Ithaca Press, Exeter, UK. 

Keay, R. W. J. 1989. Trees of Nigeria, Clarendon Press. 

Knight, T. M., Steets, J. A., Vamosi, J. C., Mazer, S. J., Burd, M., 

Campbell, D. R., Dudash, M. R., Johnston, M. O., Mitchell, R. J. 

and Ashman, T.-l. 2005. Pollen limitation of plant reproduction: 

pattern and process. Annual Review of Ecology, Evolution, and 

Systematics, 467-497. 

Lieberman, D. and Lieberman, M. 1987. Forest tree growth and dynamics 

at La Selva, Costa Rica (1969-1982). Journal of tropical ecology, 

3: 347-358. 

Newstrom, L., Frankie, G. W. and Baker, H. 1994. A new classification 

for plant phenology based on flowering patterns in lowland tropical 

rain forest trees at La Selva, Costa Rica. Biotropica, 141-159. 

Primack, R. B. 1987. Relationships among flowers, fruits, and seeds. 

Annual Review of Ecology and Systematics, 18, 409-430. 

Richards, P. 1957. Ecological Notes on West African Vegetation: I. 

The Plant Communities of the Idanre Hills, Nigeria. The Journal of 

Ecology, 563-577. 

Schwartz, M. D. 2003. Phenology: an integrative environmental science, 

Springer. 



In Vitro Evaluation of Antibacterial Activity of A Novel Polyenzyme Preparation 

Immunoseb 

ANITA JOSHI 2 , VARSHA SHAHANE 1 , VARSHA GORE 1 , ANJU KAGAL 1 , SHILPA RISBUD 2 AND RENU 

BHARADWAJ 1 

1 

B. J. Medical College, Pune, India. 2Advanced Enzyme Technologies Ltd. Thane, India and Specialty Enzymes 

Co., USA, Group of companies 

e-mail : renu.bharadwaj@gmail.com 

ABSTRACT 

Novel polyenzyme formulations which are proprietary blends 

of different enzymes such as bromelain, papain, acid and 

alkaline proteases, serratiopeptidase, lysozyme etc. have been 

developed by Advanced Enzyme Technologies Ltd., Thane and 

Specialty Enzymes Co. USA, group of Companies, Screening 

for antibacterial activity of Immunoseb, a novel polyenzyme 

formulation under development, was done using ATCC strains 

and clinical isolates of Salmonella typhi, Pseudomonas 

aeruginosa, E. coli, Staphylococcus aureus, Streptococcus pyogenes 

and S. pneumoniae employing a modified antibiotic 

susceptibility test. The polyenzyme formulation was found to 

have activity against P. aeruginosa S. aureus, S. pyogenes and S. 

pneumoniae at the test concentration. Synergistic effect was 

established when Immunoseb was used in conjunction with 

Ampicillin against E. coli and with Penicillin against 

Streptococcus pyogenes. Partial synergy was established when it 

was used in conjunction with Ceftazidime against Pseudomonas 

aeruginosa and with Ciprofloxacin against Salmonella typhi. 

Antagonism was established when it was used in combination 

with Ceftazidime against S. typhi and with Ciprofloxacin against 

both, coagulase negative staphylococcal strain and 

Staphylococcus aureus. Immunoseb has antibacterial activity 

against some commonly encountered pathogens and therefore 

has the potential of being a novel broad range antibacterial 

drug. It can be used in conjunction with those antibiotics with 

which a synergistic effect or partial synergy could be 

demonstrated since their MIC levels could be decreased. Thus 

they appear to have good potential as potentiators of currently 

used antibiotics. In cases where antagonism was established, it 

should not be used in combination with that particular 

antibiotic. 

Key words Polyenzyme formulation, novel antimicrobial, broad 

spectrum antimicrobial, MIC. 

The discovery of antibiotics in the 1940’s is regarded as 

a major milestone in medical research, one which was 

responsible for controlling morbidity and mortality of several 

diseases. The next few years saw development of different 

antibiotics leading to a rapid decline in death rate and increased 

life spans. However, over the years, the indiscriminate use of 

antibiotics has led to an increased emergence of multi drug 

resistant pathogenic strains that do not respond to the usual 

line of treatment. In their fight for survival, bacteria have 

developed their own mechanisms to confront antibiotics and 

in the process have emerged much more stronger and resilient. 

Consequently, those infections that were once curable are 

now on the verge of becoming incurable. The situation is 

such that we are now facing a threat of epidemics of resistant 

bacteria. 

Therefore, it has become imperative to search for new 

antimicrobials so as to combat this problem and provide 

effective treatment. In addition to antibiotics and chemically 

synthesized drugs, the trend to look out for alternative 

medicines such as natural or herbal medicines is increasing 

because they may have fewer side effects or toxicity and may 

be more acceptable to the body owing to their natural source. 

Being much more complex as compared with synthetic drugs 

and antibiotics, bacteria would find it difficult to build 

resistance to them. 

The following study aims at exploring the possibility of 

using a polyenzyme formulation, Immunoseb, developed by 

Advanced Enzyme Technologies Thane / Specialty Enzymes 

Co. USA, both, as an antibacterial agent or as a potentiator of 

existing antimicrobial agents. These polyenzyme formulations 

are blends of purified mixtures of enzymes obtained from 

animal glands, plants, bacteria and fungi. 


Part 1: Screening of polyenzyme formulation for 

antibacterial activity by a modified agar based antibiotic 

susceptibility test 

To test whether the polyenzyme mixture possessed any 

antibacterial properties, a modified antibiotic susceptibility 

method was used. The method involved inoculation of 

standardized bacterial cultures (matched with 0.5 Macfarland 

standard) on Mueller Hinton agar (MH agar, Qualigens Fine 

Chemicals, GlaxoSmithKline, Mumbai) so as to obtain lawn 

cultures after which 10 ml of enzyme formulation (20mg/ml in 

normal saline) was applied directly as a drop, onto the cultures 

and incubated appropriately (Miles, et. al., 1938 b). In case of 

streptococci, Blood Agar Base no. 2 (Oxoid) supplemented 

with sheep blood was used. Zones of clearing in the lawn of 

bacteria indicated antibacterial activity of the enzyme

JOSHI et. al., : In Vitro Evaluation of Antibacterial Activity of A Novel Polyenzyme Preparation Immunoseb 143 

formulation. The zones of clearance were graded as per the 

scheme given in the results section. 

Organisms used : 

For the study 5 strains of each of the 5 organisms 

mentioned below were used. Thus, a total of 25 strains were 

used for the study. These were as follows: Staphylococcus 

aureus ATCC 25923 and 4 clinical isolates Pseudomonas 

aeruginosa ATCC 27853 and 4 clinical isolates, E. coli ATCC 

25922 and 4 clinical isolates; Streptococcus pneumoniae ATCC 

49619 and 4 clinical isolates of Streptococcus pyogenes and 5 

clinical isolates of Salmonella typhi. These are the pathogens 

that are most commonly encountered in nature. Moreover, 

these are also organisms that are routinely used in a primary 

screen for a new antimicrobial (Victor, et. al., 1991). 

Part 2: Study of interaction between formulation and 

currently used antibiotic by checker board MIC method: 

The checkerboard Minimum Inhibitory Concentration 

(MIC) method using the Mueller Hinton broth (Qualigens Fine 

Chemicals, then GlaxoSmithKline Pharmaceuticals Ltd., 

Mumbai) was used to study the combined effect of currently 

used antibiotic with the enzyme formulation against organisms 

of choice (Victor, 1991) Results were recorded as positive if 

growth or turbidity/ button formation (where bacteria settle 

to form a tight pellet):could be seen and negative if no growth 

or clear medium was seen. MIC is the highest dilution where 

there is no button formation. 

Organisms used : 

E. coli ATCC 25922, P. aeruginosa ATCC 27853, S. typhi, 

Staphylococcus (coagulase negative): emerging opportunistic 

pathogen, Staphylococcus aureus, Streptococcus pyogenes 

Interpretation : 

● 

● 

● 

● 

If FIC < 0.5: synergistic effect of 2 drugs 

If FIC = 0.5-1: partial synergy or addition 

If FIC lies between 0.5-2.0: indifference 

If FIC > 2: antagonistic effect of 2 drugs 

Synergy: Synergistic action of a combination of 

antibiotics is present if the effect of the combination exceeds 

the additive effects of the individual components. 

Partial synergy/ addition: The additive effect of 

combination is equal to that of the sum of the effects of the 

individual components. 

Indifference: An indifferent effect of a combination is 

one that is equal to the effects of the most active component. 

Antagonism: Antagonism is present if a reduced effect 

of a combination of antibiotics is observed in comparison 

with the effect of the most effective individual substance. 

(Victor, et. al., 1991, Bharadwaj, 2013). 

Part 3: Studies with Streptococcus pyogenes 

An agar-based method was used to study the interaction 

between Penicillin and Immunoseb. On the first Blood agar 

base no. 2 (Oxoid) plate inoculated with a standardized 

suspension of S. pyogenes, two penicillin disks (potency 10 

units; HiMedia Laboratories, Mumbai) were placed and a drop 

of (7 ml) of 20mg/ml solution of Immunoseb was applied onto 

one of the two disks. Thus the first served as a control while 

combined effect could be observed on the second disk. On a 

second plate inoculated with S. pyogenes, a penicillin disk 

was placed along with another disk impregnated with 7 ml of 

20mg/ml Immunoseb at a distance of 1.5cm from the penicillin 

disk. Similarly, a disk impregnated with 7 ml of another 

formulation under development was placed at a distance of 

1.5cm from the penicillin disk Fig. 1. If the zone size of the 

combination disk is ³ 2mm larger than the largest zone size 

produced by either substance alone, synergy has occurred. 

MIC of Penicillin along with Immunoseb against S. 

pyogenes : 

Fixed concentration (40 mg/ml) of Immunoseb was added 

to serial dilutions of penicillin in glucose broth, and the tubes 

were inoculated with a standardized suspension of S pyogenes. 

After appropriate incubation, all the suspensions were 

streaked onto Blood Agar plates ad incubated appropriately. 


Part 1: Screening for antibacterial activity of novel 

polyenzyme formulation using modified antibiotic 

sensitivity method: 

For recording the results, the following scheme was used: 

Clear zone 3+ 

< or equal to 5 colonies in the zone 2+ 

> or equal to 5 colonies in the zone 1+ 

Lawn culture with no zone 0 

Gradation of zones: 

Interpretation: 0 and 1+ were considered as negative or 

resistant to the formulation. 

The results obtained were as follows: 

1. Staphylococcus aureus : 

ATCC strain: 3+ 

4 Clinical isolates: 3+ 

2. Pseudomonas aeruginosa 


4 Clinical isolates (including 2 MDR strains): 3+ (See 

Figure 1). 

3. E. coli 


3 Clinical isolates: ‘0’, 1 showed 1+ zone


4. Salmonella typhi 

3 clinical isolates: ‘0’; 2 showed 1+ gradation 

5. Streptococcus pyogenes: all 4 clinical isolates 3+ 

Streptococcus pneumoniae ATCC 49619: 3+ zones 

The polyenzyme preparation, Immunoseb shows broadspectrum 

antibacterial properties. The standard and sensitive 

ATCC strains viz., Staphylococcus aureus ATCC 25923, 

Pseudomonas aeruginosa ATCC 27853 and Streptococcus 

pneumoniae ATCC 49619 are clearly sensitive to Immunoseb 

at the test concentration of 20 mg/ml, as was evident from the 

clear zones obtained. E. coli ATCC 25922 and S. typhi isolate 

used as a standard sensitive strain produced zones of inhibition 

which had >5 colonies in the zones. This shows that the two 

strains are resistant at 20mg/ml concentration to Immunoseb. 

All 4 clinical isolates of S. aureus were found to be 

sensitive to the formulation at the 20mg/ml concentration 

producing distinct and clear zones. 

All 4 clinical isolates of P. aeruginosa were found to be 

sensitive to Immunoseb at the 20mg/ml concentration. Two 

multi drug resistant strains of P. aeruginosa were found to be 

sensitive to Immunoseb at 20mg/ml concentration. (fig. 1) 

Fig. 1. 

Action of polyenzyme formulation against two strains 

of Pseudomonas aeruginosa. 

All 4 clinical isolates of S. pyogenes were found to be 

sensitive against the formulation at the 20 mg/ml 

concentration producing distinct zones. 

All 4 clinical isolates of E. coli and S. typhi produced 

zones with >5 colonies (1+ gradation) or no zones at all 

indicating resistance to Immunoseb at the 20 mg/ml 

concentration. 

Part 2: Checkerboard MIC: With the help of the formula, 

FIC values were calculated and the interaction between 

antibiotic and polyenzyme formulation was classified as 

“synergy”, “antagonism” or “partial synergy” (Table 1). 

MIC values of the formulation could be obtained for all 

the strains studied. Synergistic effect was obtained when 

Immunoseb was used in combination with Ampicillin against 

E. coli ATCC 25922. Partial synergy was observed when 

Immunoseb was used in combination with Ceftazidime against 

Pseudomonas aeruginosa ATCC 27853. Antagonistic effect 

was observed when Immunoseb was used in combination 

with Ceftazidime against Salmonella typhi. Here, the MIC of 

Ceftazidime was increased although the MIC of polyenzyme 

decreased. However independently Immunoseb is effective 

against Salmonella typhi. Partial synergy was observed when 

Immunoseb was used in combination with Ciprofloxacin 

against Salmonella typhi. In this case the MIC of Ciprofloxacin 

was lowered but that of Immunoseb remained the same. 

Independently, Immunoseb has MIC = 1.25mg/ml against this 

clinical isolate of Salmonella. 

In case of Staphylococcus aureus for the checker board 

titrations, two Staphylococcus strains were used: coagulase 

–ve strain which is being considered as an emerging 

opportunistic pathogen and a sensitive Staphylococcus 

aureus (MSSA) clinical isolate. Both strains showed 

antagonism with Ciprofloxacin. 

Part 3: Studies with Streptococcus pyogenes: Synergy 

could be established by all methods employed between 

Penicillin and Immunoseb as seen by enhanced zone of 

inhibition (Fig. 2) against S. pyogenes. The MIC of penicillin 

dropped from 0.12 mg/ml. to 0.031 mg/ml as seen by growth 

on the blood agar plates, confirming synergistic effect of 

Immunoseb with penicillin against S. pyogenes. A four-fold 

decrease in MIC value of penicillin was established clearly 

indicating synergy. 

Table1. 

Interaction between antibiotics and polyenzyme formulation 

Organism 

Antibiotic MIC of antibiotic (g/ml) MIC of enzyme FIC Interpretation 

(mg/ml) 

value 

Alone Combination Alone Combination 

E. coli ATCC 25922 Ampicillin 4 1 20 5 0.5 Synergy 

P. aeruginosa ATCC 28753 Ceftazidime 4 2 10 5 1.0 Partial synergy 

S. typhi Ceftazidime 0.5 1.0 2.5 1.25 2.5 Antagonism 

S. typhi Ciprofloxacin 2 0.5 1.25 1.25 1.25 Partial synergy 

Staphylococcus (coagulase –ve) Ciprofloxacin 0.25 2 0.62 1.25 10.0 Antagonism 

Staphylococcus aureus (coagulase Ciprofloxacin 0.5 2 0.31 0.15 4.48 Antagonism 

+ve) 

Streptococcus pyogenes Penicillin Disc Diffusion 

Method 

Synergy

JOSHI et. al., : In Vitro Evaluation of Antibacterial Activity of A Novel Polyenzyme Preparation Immunoseb 145 

The results indicate that Immunoseb has broad range 

antibacterial properties. The studies clearly show that 

Immunoseb is effective against several commonly encountered 

pathogens such as Staphylococcus aureus, Streptococcus 

pneumoniae, S. pyogenes, Pseudomonas aeruginosa, E. coli 

and S. typhi. 

Fig. 2. 

Enhanced action of Penicillin due to Immunoseb and 

another polyenzyme formulation under development 

against Streptococcus pyogenes 

Furthermore, synergy or partial synergy of Immunoseb 

with currently used antibiotic could be established in all cases 

except with the staphylococci. Therefore, Immunoseb can be 

used for potentiating the effect of antibiotics. Thereby it could 

help reduce MIC of the currently used antibiotic. This is a 

significant finding, especially in case of S. typhi. Ciprofloxacin 

is the currently used drug against S. typhi. However, increase 

in MIC of Ciprofloxacin by the currently circulating strains of 

S. typhi has become a cause for concern among clinicians. 

Lowering the MIC of Ciprofloxacin when used in conjunction 

with Immunoseb will help control borderline resistant 

Salmonella typhi infections. Such studies were carried out 

with the hope that combination therapy would prevent the 

development of resistance and allow the use of lower drug 

doses thereby reducing the risk of toxicity associated with 

one or both agents in the combination. Earlier reports show 

that enzymes have been used for combination therapy. The 

combination of trypsin and bromelain given together with 

antibiotics was found more effective for treating urinary tract 

infections than those patients who received only the antibiotic. 

This indicates a synergistic effect of the enzymes when given 

together with antibiotics. Other studies have shown that the 

level of antibiotic in target tissues could be increased by 

administering proteolytic enzymes in conjunction with 

antibiotics. For instance, such an effect was seen when 

serratiopeptidase was administered along with the antibiotic 

Cefotiam to lung cancer patients undergoing thoracotomy. 

Thus due to their digestive nature, enzymes can bring about 

better penetration of the antibiotic thereby increasing efficacy 

of action of the drug given along with it (Koyama, et. al.,). 

By itself, Immunoseb shows activity against coagulase 

negative Staphylococcus strains, which are considered to be 

emerging opportunistic pathogens. It also shows activity 

against Methicillin sensitive S. aureus clinical strains. This 

finding is of great importance since Staphylococcus is a 

causative agent of nosocomial infections of increasing 

importance. However, it should not be used in conjunction 

with Ciprofloxacin since antagonistic effect was recorded. 

Streptococci spp. are sensitive to the formulation as 

shown by the agar diffusion method. Therefore, it has potential 

applications in topical microbicidal mouthwashes, dental 

preparations, cough syrups and tablets etc. Synergy could 

be demonstrated by the agar diffusion method between 

Penicillin, which is the antibiotic of choice and Immunoseb 

against S pyogenes. 

Another important finding is the sensitivity of 

Pseudomonas aeruginosa to Immunoseb. It also is synergistic 

with third generation cephalosporins against Pseudomonas 

aeruginosa. The organism Pseudomonas aeruginosa is a 

major opportunistic pathogen responsible for hospital-based 

infections often resulting in treatment failures due to its drug 

resistance. Therefore, Immunoseb has great potential in topical 

applications for treating burn patients, and could be used in 

conjunction with currently used antibiotics. 

In conclusion, it can be said that Immunoseb shows 

good promise as a potentiator of currently used antibiotics 

against important pathogens. Besides it also shows its own 

antibacterial activity. Further to these in vitro studies, in vivo 

pharmacokinetic studies need to be undertaken in which 

additional data on the antimicrobial activity of the formulation 

can be obtained. 


Bharadwaj, R. Vidya, A. Dewan, B. and Pal, A. 2013. An in vitro study to 

evaluate the synergistic activity of norfloxacin and metronidazole. 

Ind J Pharmacol. 35: 220-226. 

Koyama A, Mori, J., Tokuda, H. 1986. Augmentation by serrapeptase 

of tissue permeation by Cefotiam (Japanese). Jpn. J. Antibiot, 39(3): 

761-771. 

Miles, A. A., Misra, S. S., Irwin, J. O. 1938. The estimation of the 

bactericidal power of the blood J. of Hyg., 38: 732-749. 

Victor Loriane 1991. Antibiotics in Lab Medicine, 3rd edition. 

a) Chapter 2: Disk Susceptibility test. J F Acar and F W Goldstein. 

pp 17-52. 

b) Chapter 21: Evaluation of new antimicrobials in vitro and in 

experimental animal infections. R Cleeland and E Squires. pp. 739-786. 

c) Chapter 13: Antimicrobial Combinations. G M Eliopoulos, R C 

Moellering Jr. pp. 432-492. 



Comparative Analysis of Morphological and Molecular Diversity in Mungbean 

(Vigna radiata L. Wilczek) 

G. ROOPA LAVANYA 1* AND SHIRISH A. RANADE 2 

1 

Department of Genetics and Plant Breeding, Allahabad School of Agriculture, Sam Higginbottom Institute of 

Agriculture, Technology and Sciences, Deemed-to-be-University, Allahabad 211 007 

(e-mail: lavanya.roopa@gmail.com) 1 

2 

PMB (Genomics), CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow 226 001 

(e-mail: shirishranade@yahoo.com) 2 

ABSTRACT 

Estimation of genetic diversity in a crop is a prerequisite for its 

improvement. Fifty four accessions of mungbean were compared 

through RAPD profiles and morphological traits. Field data of 

consecutive years was subjected to multivariate analysis and 

non hierarchical Euclidean analysis. The 54 accessions were 

grouped into 10 and five clusters on the basis of morphological 

and RAPD data respectively. The clusters, though not congruent 

to each other, revealed interesting trends for RAPD data and 

mean performance under the 10 morphological traits. Plants 

with above average performance for plant height and number 

of seeds per pod clustered together in RAPD analysis and were 

in general the superior performing accessions. Among 

morphological traits, the maximum Euclidean distance was 

between clusters IV and VIII, in good agreement with four out 

of the five RAPD data clusters. Thus by combining the RAPD 

and Euclidean analyses it would be possible to compile a panel 

of contrasting parental accessions for a meaningful mungbean 

improvement program. 

Key words Distances, Genetic diversity, mungbean, Neighbour- 

Joining dendrogram, Non-hierarchical Euclidean cluster analysis. 

Low productivity of Mungbean crop has been attributed 

to narrow genetic base (resulting in low yield potential and 

susceptibility to biotic and abiotic stresses) as well as to lack 

of suitable plant types for different cropping conditions 

(Dikshit, et al., 2009). The genetic reconstruction of plant type 

is therefore, required for developing high yielding varieties 

(incorporating and improving yield component traits) suitable 

for cultivating in several different agro-climatic regions of the 

country. The crosses between parents with maximum genetic 

divergence are generally the most responsive for such genetic 

reconstruction and improvement (Arunachalam 1981). The 

available germplasm serves as the most valuable natural 

reservoir for providing the required plant attributes for 

obtaining the high yielding crop varieties (Hawkes 1981). In 

order to utilize such accessions with maximum required plant 

attributes, it is necessary to screen and characterize the 

germplasm for the nature and extent of genetic diversity 

included in it. Characterization and cataloguing of germplasm 

have been traditionally carried out using morpho-agronomic 

traits while in the last two decades or so the molecular markers 

have also been used for germplasm characterization. The 

molecular markers, highly heritable and available in large 

numbers are often polymorphic enough to enable 

discrimination of closely related genotypes. Of the several 

molecular methods possible, using Random Amplified 

Polymorphic DNA (RAPD) profiles (Williams, et al., 1990) 

offers a rapid and reliable identification and characterization 

of genotypes by generating markers for comparative analysis 

that are quick, easy to use, free from environmental influences, 

unlimited in number, random but wide coverage of genome 

and have a relatively higher level of polymorphism (Newbury 

and Ford-Lloyd 1993). Studies have been carried out earlier 

using RAPD profiles alone or in combination with ISSR profiles 

(Bisht, et al., 1998; Santalla et al. 1998; Lakhanpaul, et al., 

2000; Afzal, et al., 2004 and Lavanya, et al., 2008) in mungbean. 

The present study was carried out to assess relationship, if 

any, between the molecular and morphological traits in a set 

of mungbean accessions representing a number of local land 

races and varieties released by the different centers with an 

objective to select divergent parents for further use in 

improvement of mungbean. 


Plant material, Morphological characterization: Fifty 

four mungbean accessions representing diverse agro ecologies 

were selected for the present study. The accessions were 

obtained from different locations of India and from Asian 

Vegetable Research and Development Centre (AVRDC), 

Taiwan (Table 1). The morphological data were generated by 

evaluating the germplasm during kharif seasons over the past 

5 years at Field Experimentation Centre, Department of 

Genetics and Plant Breeding, SHIATS, Allahabad. The 

experiment was laid in randomized complete block design in 

three replications of plot size 1x1 m 2 with 30cm and 10cm interand 

intra- row spacing, respectively. Ten plants were randomly 

selected to record the data on 10 morphological traits viz., 

plant height (cm), number of primary branches, number of 

clusters per plant, number of pods per cluster, number of pods 

per plant, number of seeds per pod, pod length (cm), days to 

maturity, seed index (g) and seed yield per plant (g). The 

observations on days to 50% flowering and maturity were 

recorded on plot basis. The pooled data was subjected to

LAVANYA & RANDADE : Comparative Analysis of Morphological and Molecular Diversity (Vigna radiata L. Wilczek) 147 

Table 1. 

Mungbean accessions used for Euclidean and 

RAPD analysis 

Sample No. Name of the accession Source 

MB1 PDM 84-139 IIPR, Kanpur 

MB2 Narendra Mung 1 NDUAT, Faizabad 

MB3 LGG 478 RARS, Guntur, AP. 

MB4 EC 30400 AVRDC, Taiwan 

MB5 MSO 9 AVRDC, Taiwan 


MB7 IPRM 90 IIPR, Kanpur 

MB8 T1 IIPR, Kanpur 

MB9 Pusa Bold 2 IARI, New Delhi 

MB10 LM 497 PAU, Ludhiana 

MB11 OUM 11-5 OUAT, Bhubaneswar 

MB12 PDM 84-143 IIPR, Kanpur 

MB13 HUM 1 BHU, Varanasi 

MB14 V 557 AVRDC, Taiwan 

MB15 LLR 1 Local Land Race 1 

MB16 ML 287 PAU, Ludhiana 

MB17 V 4512 AVRDC, Taiwan 

MB18 AKM 9601 PKV, Akola 

MB19 Lam M2 RARS, Guntur, AP. 



MB22 MH 309 PAU, Ludhiana 


MB24 PDM89-226 IIPR, Kanpur 

MB25 K851 IIPR, Kanpur 


MB27 PS 10 IARI, New Delhi 


MB29 K 2192 Kanpur Local germplasm 

MB30 PUSA 9332 IARI, New Delhi 

MB31 MUM 1-1 CCSMU, Meerut 

MB32 K 1284 Kanpur Local germplasm 


MB34 NARP 280 PAU, Ludhiana 

MB35 DMG 1103 Kanpur Local germplasm 

MB36 TARM 1 BARC, Mumbai/ PKV, Akola 

MB37 WGG 37 RARS, Warangal, AP. 

MB38 SUJATA OUAT, Bhubaneshwar 

MB39 PIMS-11/99 PAU, Ludhiana 


MB41 PDM 11 IIPR, Kanpur 

MB42 DMG 1098-1 Kanpur Local germplasm 

MB43 MARP 280 PAU, Ludhiana 

MB44 AKM 8802 PKV, Akola 

MB45 TARM 2 BARC, Mumbai/ PKV, Akola 



MB48 PM 9001 Kanpur Local germplasm 

MB49 OBGG 11 OUAT, Bhubaneshwar 


MB51 MGG 47 RARS, Madhira 


MB53 PDM 1 IIPR, Kanpur 


OUT 

Bauhinia purpurea L. NBRI, Lucknow 

multivariate analysis as suggested by Mahalanobis, 1936 and 

accessions were grouped into different clusters based on 

Euclidean distances by non- hierarchical cluster analysis 

(Spark, 1973). 

DNA isolation: Young leaf tissue from plants raised in 

the field was used for DNA extraction. It was harvested from 

the plants, washed free of dust and then quickly mopped dry 

on blotting sheets. The leaves were de-ribbed and wrapped in 

tissue paper. The packed leaf tissue was kept in Ziplock bags 

along with fine mesh blue silica gel for rapid dehydration. The 

dried leaf tissue was used for extracting the genomic DNA 

extraction. Genomic DNA was isolated using Plant DNA 

isolation kit (Bangalore Genei, Bangalore, India). Bauhinia 

purpurea L. was randomly selected as the non-Vigna 

leguminous out-group for all studies to test whether our 

reaction conditions were optimized to resolve it as distinct 

from the rest of the mungbean accessions. 

RAPD primers, reactions and agarose gel 

electrophoresis: The PCR conditions, primers used and 

agarose gel electrophoresis conditions were as reported 

previously by Lavanya, et al., 2008. 

Scoring and analysis of bands: Clear and well marked 

bands were coded in a binary form by denoting ‘0’ and ‘1’ 

intended for absence and presence of bands, respectively in 

each accession and these data then were used as input for 

further calculations. In order to describe genetic relationships 

among the mungbean accessions, RAPD band data were used 

to estimate genetic distances, based on Jaccard coefficients 

computed using the Free Tree (ver. 0.9.1.50) program (Pavlicek, 

et al., 1999). Cluster analysis was carried out based on genetic 

distances using Neighbour Joining (NJ) program in the Free 

Tree package. The resulting clusters were represented as a 

dendrogram and viewed and printed in the program Tree View 

(ver. 1.6.5; Page 2001). The AMOVA and PCoA analysis were 

carried out using GenAlEx software (Peakall and Smouse, 2012) 

by considering the mungbean accessions in five groups as 

resolved by the NJ dendrogram for the RAPD data. 


Cluster formation based on non-hierarchical Euclidean 

analysis of field data on all traits generated 10 clusters, Cluster 

V had 11 accessions, followed by cluster I with nine accessions 

while, cluster IV included only one accession i.e., EC-398889. 

The accessions developed at or released from same or different 

locations were not always grouped into the same cluster. Four 

varieties developed at Regional Agricultural Research Station, 

Lam, Guntur, Andhra Pradesh was grouped into cluster VIII 

and one variety (LGG 478) in cluster X. Similarly, three varieties 

namely PDM 84-139, PDM 11 and PDM 1 from IIPR, Kanpur 

distributed in cluster V but the other two varieties, also from 

same source, PDM 84-143 and PDM 89-226 were included in 

cluster VII and IX, respectively. Interestingly clusters III, IX 

and X comprised varieties developed at different locations


Table 2. 

Intra-cluster (diagonal shaded cells) and inter-cluster average distances (D 2 ) for 10 traits in mungbean with the 

highest values of distances indicated in bold 

Cluster No. I II III IV a V VI VII VIII IX X 

I 2.880 11.820 4.486 91.413 8.556 7.801 24.780 28.558 10.916 29.149 

II 2.554 7.290 91.394 8.744 4.844 15.682 15.809 6.760 8.375 

III 2.329 85.933 5.058 6.497 10.530 14.478 9.157 15.856 

IV a 0.000 a 73.943 86.583 104.04 70.560 87.984 83.466 

V 2.088 3.968 15.666 9.647 9.616 13.126 

VI 3.320 20.857 15.194 11.069 16.443 

VII 3.010 15.531 12.397 12.278 

VIII 5.988 21.988 11.089 

IX 3.280 12.103 

X 2.772 

a 

The cluster IV had just a single accession and hence no Intra-cluster distance could be computed. 

clusters IV and VII (104.04) followed by clusters I and IV 

(91.413) and clusters II and IV (91.394). Clusters with maximum 

inter cluster distance were found to be highly divergent 

groups. Recombination among accessions of diverse clusters 

is necessary to obtain desirable plant types. Hence inter 

cluster distance must be taken into consideration while 

selecting the parents for hybridization program. 

Table 3. 

Mean performance for 10 characters of mungbean 

accessions included in five clusters as in NJ 

dendrogram (Figure 1) by RAPD analysis 

Character (Mean value) NJ dendrogram cluster number 

I IIA IIB IIC IID 

Plant height (47.65 cm) 58.87 40.5 42.56 49.27 58.16 

No. of primary branches (3.66) 3.79 3.33 3.56 3.58 3.95 

No. of clusters / plant (15.41) 15.73 14.83 4.48 13.75 17.67 

No. of pods / cluster (3.29) 3.67 3.11 3.37 3.19 3.25 

No. of pods / plant (40.57) 45.91 36.83 40.19 34.38 48.47 

No. of seeds / pod (9.65) 10.12 7.78 9.85 9.24 9.58 

Pod length (6.57 cm) 7.22 5.82 6.50 6.85 7.27 

Days to maturity (77.39) 77.61 73.00 75.33 77.31 78.26 

Seed index (3.82 g) 3.92 3.13 3.95 3.42 3.81 

Seed yield / plant (11.04 g) 13.96 9.47 9.82 10.96 12.90 

Fig. 1. 

NJ tree for the RAPD band data in case of the 

mungbean accessions and outgroup DNA (indicated by 

the underscored label OUT) and its separation from the 

mungbean accessions is indicated by a thick line. The 

OTU labels are to the right of the branches and are as 

given in Table 1. The branch lengths are based on the 

distances between the accession pairs. The distribution 

of the accessions into different clusters is indicated by 

oval labels on the corresponding branches of the 

dendrogram. 

(Table 2). Cluster VIII exhibited maximum intra-cluster distance 

(D 2 ) and the accessions of this cluster were diverse (Table 2). 

Inter-cluster distance (D 2 ) was found maximum between 

High (>20%) genotypic coefficient of variation (GCV) 

and phenotypic coefficient of variation (PCV) were recorded 

for plant height, number of clusters plant -1 , number of pods 

cluster -1 , number of pods plant -l , seed index and seed yield 

plant -1 . High genotypic and phenotypic coefficient of variation 

for seed yield plant -l , number of branches plant -l and number 

of pods plant -1 were also reported earlier by Reddy 1997 and 

Loganathan et al., 2001 while for 100-seed weight by Samad 

and Lavanya 2005 reported a high genotypic and phenotypic 

coefficient. A scrutiny of Table 5 revealed that all characters 

recorded moderate (30-60%) to high (above 60%) heritability 

except number of primary branches. Burton 1952 has suggested 

that genetic variation along with the heritability estimates 

would give a better idea about the expected efficiency of 

selection. Thus, a character possessing high GCV along with 

the high heritability will be valuable in a selection program. 

Seed yield plant -1 and number of pods plant -1 recorded high 

estimates of GCV coupled with high heritability in the present


study. Maximum genetic advance was recorded for plant height 

followed by number of pods plant -1 . However, high (30%) 

estimates of genetic gain were registered for plant height, 

number of clusters plant -1 , number of pods cluster -1 , number 

of pods plant -1 , pod length and seed yield plant -1 . Low genetic 

advance as per cent of mean was observed for days to maturity, 

indicating the involvement of non additive gene action in 

controlling this trait and heterosis breeding may be useful for 

further generation of variability for this character. Heritability 

estimates along with genetic advance are more useful than 

heritability alone in predicting the effectiveness of selection. 

Further, the heritability estimates coupled with genetic gain, 

indicate the mode of gene action in choosing an appropriate 

breeding methodology. High genetic advance as per cent of 

mean coupled with high heritability recorded for plant height, 

seed yield plant -1 and number of pods plant -1 . This situation 

indicates that the genetic variances for these traits are 

probably owing to their high additive gene effects (Johnson, 

et. al., 1955) and thus there is better scope for improvement of 

these traits through direct selection. 

Significant morphological variation was present among 

the accessions for all traits studied. Fifty four accessions were 

distributed into 10 clusters and distribution of accessions 

into different clusters, suggested the presence of substantial 

genetic divergence among the germplasm lines screened in 

the present investigation and further, indicated that this 

material may serve as good source for selecting the diverse 

parents for hybridization program, aimed at isolating desirable 

recombinants for seed yield as well as other traits (Naidu and 

Satyanarayana, 1991 and Manivannan, et. al., 1998). Choice 

of parents plays a vital role in hybridization program. Best 

performing accessions from distant clusters like IV, VII, I and 

II in the present study should be selected for further breeding 

program. Progenies of genetically diverse parents are likely to 

produce a broad spectrum of variability in segregating 

generations which will facilitate the isolation of transgressive 

segregants in a hybridization program. 

The perusal of clustering pattern of the accessions 

clearly revealed the lack of relationship between geographic 

distribution and genetic diversity as the distribution of 

accessions into various clusters was fairly random (Reddy, 

1997 and Loganathan, et. al., 2001). The grouping of 

accessions originating from different eco-geographical regions 

into one cluster could be attributed to frequent exchange of 

breeding material and due to operation of similar forces of 

natural and artificial selection resulting in perpetuation, 

adaptation and stabilization of similar accessions (Murty and 

Arunachalam, 1966). In case of mungbean, Singh, et al., 2010 

have estimated genetic variability in over 200 hundred varieties 

by Euclidean hierarchical analysis and have reported a wide 

diversity among the germplasm that resulted in grouping of 

all the accessions into eight clusters. However, they have 

also observed that genetic diversity and geographical origin 

of the accessions were not congruent. 

Fig. 2. The Principal Coordinates (PCoA) plot for all the 54 

mungbean accessions considered as five groups as in NJ 

dendrogram (Fig. 1) are indicated by colour coded 

symbols as shown in the figure. The numbers by each 

symbol correspond to the sample numbers as in 

Table 1. 

In the present study, data from seven primers (174 bands 

with an average of 24.84 bands per primer) were used for the 

computation of NJ dendrogram and for AMOVA and PCA 

analysis as described. In the pairwise distances measure for 

computation of the NJ dendrogram, the lowest distance was 

0.524 between the accessions, LLR 1 and ML 287 while the 

accessions WGG 37 and PDM 1 and WGG 37 and OBGG 52 

exhibited the greatest distance 1.0 from each other (data not 

shown). The NJ dendrogram (Fig. 1) distributed all 54 

mungbean accessions into two broad clusters, I (with 11 

accessions) and II (with remaining 43 accessions). The latter 

cluster was further composed of four subclusters (II A with 

six, II B with nine, II C with 15 and II D with 13 accessions), as 

shown in Figure 1. The out group genotype, B. purpurea was 

clearly distinguished from all mungbean accessions and 

separated from them in the NJ dendrogram. The AMOVA and 

PCoA are analyses are shown in Figure 3. 

The accessions were arranged in order of their 

occurrence in the NJ dendrogram clusters and their 

performances for 10 characters were recorded. Cluster I plants 

were found to be above average for all 10 characters; Cluster 

IIA plants were distinctly below average for all 10 characters; 

Cluster IIB plants were above average pods/ cluster, seeds/ 

pod and seed index. Cluster IIC and IID plants were mostly


medium performers but distinguished from each other in that 

cluster IIC plants show greater plant height, fewer days to 

maturity and higher seed yield per plant than the plants of 

cluster IID (Table 3). Plants of cluster I in general show superior 

performance in most of the traits examined except days to 

maturity where more than half the plants have below average 

performance. Such a distribution of plants based on agronomic 

performance and supported by RAPD proûle analyses are a 

very good starting point for further breeding eûorts involving 

contrasting parental lines. Since the plants of the cluster I are 

in general, superior performing varieties, it is these varieties 

that should be used not only for commercialization but also 

as, the starting material for further improvement by breeders. 

Random Amplified Polymorphic DNA, a method widely 

used to study genetic polymorphism revealed a wide and 

diverse genetic base of the germplasm accessions analyzed. 

Earlier, low to moderate polymorphism was observed while 

analyzing 32 Indian mungbean cultivars using 21 RAPD 

primers where genetic didstances ranged from 0.06 – 0.30 

(Lakhanpaul, et al., 2000). However, as has been stated earlier, 

the present study with a larger number of accessions 

representing more geographical regions is more efficient in 

resolving the extent of genetic diversity amongst these 

accessions. The different accessions may have common 

pedigrees (whole or partial) and may have been subjected to 

same selection during their breeding but are still 

distinguishable from each other on the basis of their RAPD 

profiles. On the basis of distances and cluster analysis, 

accessions WGG 37, PDM 1and OBGG 52 were found to be 

quite distinct and these can be used for their desirable 

characteristics in breeding programs for mungbean 

improvement. The genetic similarities obtained from the 

analysis can also be used for the selection of the parents to 

generate mapping populations and for selecting parents for 

breeding purposes. 

Clustering of cultivars into five clusters showed 

reasonable variability that may be exploited for yield 

improvement (Afzal, et al., 2004). The tree obtained by the NJ 

clustering method did not show any significant correlation 

between the genetic divergence and geographical distribution. 

In contrast, the varieties developed from different 

geographical regions grouped into same cluster. For example 

cluster II B included PS 10, TARM 1, K 1284 and ML 406 and 

II D included K 851, ML 583, AKM 9601, LGG 491. Such 

clustering of cultivars of different locations ignored the 

influence of geographic variations within the genetic diversity 

of mungbean. Such a lack of correlation between geographic 

diversity and genetic diversity in mungbean has also been 

reported by earlier workers (Bisht, et al., 1998 and 

Manivannan, et al., 1998). The overall levels of genetic 

diversity in mungbean in the present study is also supported 

AMOVA analysis where within group variations were 

approximately five-fold more than those between groups. 

These results suggest that the individuals in each population 

are likely to be as different from each other as from any other 

mungbean accession selected at random. Similarly in case of 

the PCA analysis (Figure 2), the different groups of accessions 

are not well resolved from each other, indicating overlap of 

genetic relatedness among individual accessions of the 

different groups. Thus the accessions appear to be discrete 

genotypes with very little or no common parents and 

pedigrees. Despite this, the accessions are grouped together 

for common traits (morphological as well as RAPD profiles) 

which will allow efficient screening of these genotypes for 

the presence of common alleles for use in molecular breeding 

program. RAPD markers have been used for the identification 

of cultivars and determination of the genetic relationships 

among cultivars of other leguminous crops including 

Phaseolus vulgaris (Skroch, et al., 1992), cowpea (Mignouna, 

et al., 1998) and Vigna angularis (Yee, et al., 1999). The present 

study has also shown the usefulness of the method to not 

only assess the range of genetic diversity in the germplasm. 

The present study showed that the clustering pattern of 

accessions is different when analyzed through non hierarchical 

Euclidean cluster analysis based on morphological traits and 

molecular marker (RAPD) analysis. The 54 accessions were 

grouped into 10 clusters based on Euclidean cluster analysis 

and five clusters on RAPD analysis. The environment might 

have influenced the expression of morphological traits to some 

extent, resulted in formation of more number of clusters. High 

estimates of genotypic coefficient of variation and heritability 

and genetic advance as per cent of mean indicate the 

predominance of additive gene action in controlling these 

characters and simple directional selection may be effective 

to improve these characters. The accessions MB 37, MB 42 

and MB 45 grouped in IIC cluster and MB 51, MB 52 and 

MB54 grouped into cluster IIA together were included in 

cluster I based Euclidean cluster analysis. Similarly cluster 

IID included MB 14, MB 19, MB 21 and MB 23 were grouped 

into cluster VIII based on cluster Euclidean cluster analysis. 

Further, the accessions included in cluster III (MB29 and 

MB 36) and cluster V (MB 27 and MB 28) were grouped into 

IIB based on RAPD analysis. These results suggested that 

Euclidean cluster analysis largely corresponded to molecular 

analysis. Diverse accessions observed in the present study 

could be a good alternative for successful mungbean breeding 

program. 

ACKNOWLEDGEMENT 

Authors are grateful to the Director Research, Sam 

Higginbottom Institute of Agriculture, Sciences and 

Technology, Allahabad and Director, CSIR-National Botanical 

Research Institute, Lucknow for facilitating the present study.


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Efficient Protocol for In Vitro Regeneration in Pigeonpea (Cajanus cajan (L.) Millsp.) 

TUSHITA GARGI, S. ACHARYA, J.B. PATEL AND VANDANA THAKKAR 

Centre of Excellence for Research on Pulses, S. D. Agricultural University, Sardarkrushinagar, 

385 506 (Gujarat) 

e-mail: t.gargi21@gmail.com 

ABSTRACT 

The present study precisely emphasized on the establishment 

of a reproducible regeneration protocol through different 

explants in pigeonpea [Cajanus cajan (L.) Millsp.] using 

different explants like primary leaf, epicotyls, cotyledonary 

node and embryonic axis in variety ‘GT 100’. In vitro raised 

fourteen days old seedlings grown aseptically on Murashige 

and Skoog’s medium were most efficient in producing multiple 

adventitious shoot from cotyledonary node explants and shoot 

bud originated from the petiolar cut end of explants. Efficient 

regeneration in pigeonpea from callus was obtained using MS 

medium supplemented with growth regulator viz; 5 ìM 

Gibberellins for shoot elongation, precise combination of BAP 

(22 ìM) and kinetin (10 ìM) for formation of shoot buds and 

10ìM NAA for root initiation under precise conditions. 

Key words Callus induction, Cajanus cajan, BAP–6- 

benzylaminopurine, Murashige and Skoog’s medium, Kin – kinetin. 

Pigeonpea is an important grain legume that dominates 

the vegetarian diet particularly in semi-arid tropics due to rich 

source of protein and minerals (Nene and Sheila, 1990). The 

productivity of pigeonpea has been static for the last four 

decades mainly due to narrow genetic base. The gene mining 

through conventional methods is rather difficult due to 

crossing barriers among different species. The availability of 

a genetic transformation system could precisely facilitate the 

transfer of genes across crossing barriers affecting both 

quantitative and qualitative production efficiency in 

pigeonpea (Lawrence and Koundal, 2001, Singh, et. al., 2003). 

The regeneration protocols are prerequisite for genetic 

transformation and have been vividly reported in pigeonpea 

(Naidu, et al., 1995; Geetha, et al., 1998; Mohan and 

Krishnamurthy, 1998, Venkatachalam, et al., 1999, and Guru 

Prasad, et. al., 2011). The present study precisely emphasized 

on the establishment of a reproducible regeneration protocol 

through different explants in pigeonpea. 


The present study was carried out at the Centre of 

Excellence for Research on Pulses, Sardarkrushinagar 

Dantiwada Agricultural University, Sardarkrushinagar, Gujarat, 

India using genetically pure seed of a variety having 

determinate growth habit viz., GT 100 of pigeonpea. 

Culture medium and conditions 

Seeds were soaked in tap running water for 15 minutes, 

rinsed in 70% alcohol for 10 minutes and surface sterilized 

with 0.1% aqueous mercuric chloride solution for 15 minutes 

followed by further rinsing five times with sterile distilled water. 

Five surface sterilized seeds were germinated in conical flask 

(100ml) containing 40 ml of MS basal medium containing 3 % 

sugar under16h photoperiod condition under aseptic 

condition after removing its seed coat. Fourteen days old 

seedlings were used as a source of auxiliary leaf, shoot tip, 

cotyledonary node and hypocotyls explants. Transverse 

sections of each types of explants measuring approximately 

2mm in length were cultured in culture tubes (25X150 mm) 

containing 20 ml culture medium. The explants were cultured 

on MS medium containing 3% sucrose, 0.8% extra pure agar 

powder and 0.044% calcium chloride supplemented with 

6-benzylaminopurine (5, 10, 15, 20 and 22 ìM). 

For shoot initiation twenty days old callus were 

transferred in culture tubes (25X150 mm) containing 20 ml MS 

medium containing 3% sucrose, 0.8% extra pure agar powder 

and 0.044% calcium chloride supplemented with different 

combinations of concentrations of growth regulators viz; 

6-benzylaminopurine (5, 10, 15, 20 and 22 ìM) in combination 

with Kinetin (2, 5, 7, 10 and 15 ìM) and Gibberellins (1, 2, 3, 4 

and 5 ìM). 

Thirty days old multiple shoots were transferred in 

rooting medium containing MS medium with 3% sucrose, 0.8% 

extra pure agar powder and 0.044% calcium chloride 

supplemented with different concentrations of Naphthalene 

acetic acid (2, 4, 6, 8 and 10ìM). The uniform incubation 

conditions were kept as 26 0 C under 16h photoperiod for all 

cultures. The plantlets with inducted roots were transferred 

to greenhouse for hardening. 


The experiment evinced callus initiation from the cut 

ends of the cotyledonary node, hypocotyls and shoot tip 

within 10-15 days of incubation, while no callusing was 

observed on control media. Among the various auxins and 

cytokinins studied BAP (22 ìM) and Kinetin (10 ìM) was 

observed to be highly potent for callus induction while 

cotyledonary node was most efficient among the explants 

that produced multiple adventitious shoot. Further the shoot

GARGI et al., Efficient Protocol for In Vitro Regeneration in Pigeonpea (Cajanus cajan (L.) Millsp.) 153 

bud originated from the petiolar cut end of explants. Among 

the different concentrations of Gibberellins (1, 2, 3, 4 and 5 

ìM) used for shoot elongation, 5 ìM was observed most potent 

for multiple shoot elongation. For root induction from multiple 

shoot plantlets, 10ìM of the different concentrations of NAA 

(2, 4, 6, 8 and 10 ìM) was found to be the best. 

The nodular calli obtained from distal cotyledonary 

segments on MS basal medium supplemented with BAP (22 

ìM) and 10 ìM kinetin were separated from the explants and 

transferred to MS medium supplemented with different 

concentrations of BAP (5, 10, 15, 20 and 22 ìM) and Kinetin (2, 

5, 7, 10 and 15 ìM) to evaluate the differentiation of the callus 

into shoots/shoot buds. All the cultures were incubated at 

26 0 C under 16h photoperiod for 4 weeks. The calli were 

transferred to freshly prepared medium 4 times at an interval 

of 4 weeks each. Though shoot bud initiation was observed 

in all concentration combinations, yet the formation of shoot 

buds were obtained in media with precise combination of BAP 

(22 ìM) and kinetin (10 ìM). The cotyledonary segments swell 

and turn green after 4 weeks in culture, producing small, green, 

dome-like structures (Fig. a) all over the surface of the 

cotyledonary segment. After 5 weeks of culture these 

structures developed into shoot buds (Fig. b, c, d, e and f). 

Fig. 1: 

Callus initiation (a), shoot initiation (b), shoot 

elongation (c & d) and multiple shoots development 

(e, f, g & h) from callus obtained from cotyledonary 

node explants in (Cajanus cajan (L.) Millsp.) 

The shoot buds obtained from cotyledonary segments 

on MS basal medium supplemented with BAP (22 ìM), kinetin 

(10 ìM) along with explants were transferred as a mass to test 

tubes containing various media combinations for shoot 

elongation. The elongation of shoot buds had been achieved 

when MS basal medium was supplemented with GA3 (5 ìM) 

(Fig. g and h). 

Among the different concentrations of NAA (2, 4, 6, 8 

and 10ìM) used for root initiation, 10 ìM NAA revealed the 

highest rooting. Thus, at regeneration in pigeonpea from 

callus can be had by using MS medium supplemented with 

growth regulator viz; 5 ìM Gibberellins for multiple shoot 

elongation, precise combination of BAP (22 ìM) and kinetin 

(10 ìM) for formation of shoot buds and 10 ìM NAA for root 

initiation under precise conditions. 


Geetha, N. Venkatachalam, P. Prakash, V. and Lakshmisita, G. 1998. 

High frequency induction of multiple shoots and plant regeneration 

from seedling explants of pigeonpea (Cajanus cajan L.) Current 

Science, 17:1036–1041. 

Guru Prasad, M. Prasad, T.N.V.K.V. and Sudhakar, P. 2011. In vitro 

proliferation of shoot regeneration from embryo of Cajanus cajan 

L (var.LGG-29). Journal of Developmental Biology and Tissue 

Engineering, 3(5): 62-65. 

Lawrence, P.K. and Koundal, K.R. 2001. Agrobacterium 

tumefaciansmediated transformation of pigeonpea (Cajanus cajan 

(L.) Millsp,) and molecular analysis of regenerated plants Current 

Science 80: 1428-1432. 

Mohan, M.L. and Krishnamurthy, K.V. 1998. Plant regeneration in 

pigeonpea [Cajanus cajan (L.) Millsp.] by organogenesis. Plant 

Cell Report, 17:705–710 

Naidu, R.B., Kulkarni, D.D. and Krishnamurthy, K.V. 1995. 

Genotypedependent morphogenetic potentiality of various explants 

of a food legume, the pigeonpea (Cajanus cajan L.). In Vitro Cellular 

& Developmental Biology. Plant 31: 26–30. 

Nene, Y.L. and Sheila, V.K. 1990. Pigeonpea: geography and importance. 

In: The pigeonpea eds. Nene Y.L, Hall S.D., Sheila, V.K. CAB, 

Wallingford, UK, pp. 1–14. 

Singh, N.D., Sahoo, L. Sonia and Jaiwal, P.K. 2003. In vitro shoot 

organogenesis and plant regeneration from cotledonary node and 

leaf explants of pigeonpea (Cajanus cajan (L.) Millsp.), Molecular 

Breeding, 11: 159-168. 

Venkatachalam, P. Geetha, N. Khandelwal, A. Shaila, M.S. and Lakshmi 

Sita, G. 1999. Induction of direct somatic embryogenesis and plant 

regeneration from mature cotyledon explants of Arachis hypogaea 

L. Current Science, 77: 269-273. 



Study on Two Edible Spiders of the Genus: Nephila (Fam. Nephilidae) of Manipur, 

India 

1 

A.KANANBALA, 2 M. BHUBANESHWARI AND MANJU SILIWAL 1,2 

Entomology Research Laboratory, P.G. Department of Zoology,D.M. College of science Imphal 795 001 

Wildlife Information Liaison Development Society, 9-A, Lal Bahadar Colony, Peelamedu, 

Coimbatore 641 004, Tamil Nadu, India 

e-mail: akhamkanan@gmail.com 1 

ABSTRACT 

This paper deals with an account of two edible spiders belonging 

to the genus Nephila (Family: Nephilidae Simon, 1894) from 

Manipur , India. The two species are N. pilipes Fabricus 1793 

and N. clavata L. Koch 1878. These two species are reported for 

the first time from Manipur. 

Key words Nephila pilipes, Nephila clavata Manipur, edible spider 

The family Nephilidae Simon, 1894 includes long-legged 

orb weave spiders. These orb-weavers occupy a variety of 

habitats and are important predators of large arthropod prey 

in most tropical and subtropical ecosystem worldwide 

(Sebastian and Peter, 2009). N. maculata (Fab.) were found to 

be one of the most effective bioagents to control the insect 

pest of vegetable crops of Eastern Himalayas (Satpathi, 2004). 

They are easily identified by their long legs and colourful. 

Females are larger than males. The genus Nephila Leach 1815 

are commonly known as Banana spider, containing 27 different 

species.Several species of banana spider are eaten in New 

Guinea after roasting over open fire. In Austria and Caledonia, 

N. edulis is also eaten. A total of 6 species of this genus 

Nephila are reported from India so far. 

In Manipur two species of Nephila, N. clavata and N. 

pilipes are commonly found in the forest of hill and nearby 

adjoining valley areas. The local people after collecting 

buckets ful, smoked and sometimes dried in the pan in open 

fire and a popular dish was made. 

The present paper contains the description of two edible 

spider species, N. clavata , N. pilipes found in the different 

hill districts as well as small hillock area of valley district of 

Manipur. 


Altogether 5@&N. pilipes and 5@& and 1 B& of N. 

clavata were collected from different districts of Manipur and 

studied in the Entomology Research Laboratory of Zoology 

Department; D.M. College of Science, Imphal. The collected 

specimens are kept in 70% alcohol, photographed and 

deposited in the Entomology Research Laboratory Museum 

of Department of Zoology, D. M. College of Science. 

Morphometry of the spider was taken with vernier caliper and 

ocular meter. All measurements are in millimeter. 


Nephila pilipes Fabricius. 1793 [Fig. 1 and 2] 

Aranea pilipes Fabricius, 1793, Ent.sys., 2:407-428 

Description of female from Ukhrul District. 

Total length- 27.47, Cephalothorax- 8.80 long, 7.02 wide, 

Abdomen- 18.67 long, 8.41 wide. 

Colour in alcohol : Carapace grayish-black, cheliceraldark 

brown, labium brown with yellowish distal margin, maxilla 

dark brown with yellowish margin. Sternum dark brown, legs 

dark brown, palp dark , ventral femur to patella light yellow, 

tibia and tarsus black. Abdomen- Dorsum yellowish in the 

middle with lateral brownish colour and ventrum with yellow 

and dark brown spots. 

Cephalothorax : Greyish black, longer than wide, slightly 

narrower in front, cephalic region higher than thoracic region 

and provided with a pair of sharp tubercles posteriorly. 

Thoracic region flat with a deep circular fovea. Clypeus 0.55 

long. 

Eyes : Eight eyes in two rows, both rows recurved. Ocular 

region 3.68 long, 0.91 wide, median eyes equal in size, lateral 

eyes small and subequal in size, close and situated on 

prominient tubercles. Maxillae :2.88 long, 1.71 wide. Elongated, 

broader distally, dark brown with yellowish outer margins, 

provided with distinct scopulae. Labium : 1.72 wide, 1.37 long, 

deep brown with yellowish distal margin. Chelicera :5.28 long, 

strong, stout darkbrown with 4 retromarginal and 3 promarginal 

teeth respectively. 

Legs : Legs very long, strong, clothed with hairs and 

spines; coxae of legs and proximal half of palpus yellowish in 

colour ventrally. All legs are with 2 claws. Pedipalp provided 

with trichobothria on the lateral side in all the segments. Leg 

formla 1423. 

Spination : 

Abbreviation: fe = femur, pat = patella, tib = tibia, P = 

prolateral, met = metatarsus, r = retrolateral, d=dorsal v= 

ventral, mm = millimeter. 

Leg I: Fe- p=12, r=10, pat-r=1, ti- p=2, r=6, a=2, mt- p=18, 

r=13, Leg II: Fe- p=3, r=6, ti- p=1, r=6, mt- p=11, r=5. Leg III: Fep=2, 

r=2, pat- r=1, ti- p=3, r=3, mt- p=7, r=6. Leg IV: Fe- p=3, 

r=2, pat- p=2, ti- p=5, r=3, mt- p=7, r=2.

Kananbala et. al., : Study on two edible spiders of the Genus: Nephila (Fam. Nephilidae Simon 1894) of Manipur, India 155 

Measurements of legs segments 

Leg Femur Patella Tibia Metatarsus Tarsus Total 

I 17.10 3.10 13.85 21.45 3..80 59.30 

II 14.95 2.85 10.75 16.95 3.20 48.70 

III 9.40 2.25 5.60 9.35 2.60 29.20 

IV 17.25 2.90 11.30 16.80 3.85 52.10 

Leg formula 1423 

Pedipalp 3.51 1.48 1.95 - 3.58 10.52 

Abdomen : Abdomen long, cylindrical, clothed with hairs, 

slightly over lapping on the cephalothorax. Dorsum with a 

pair of mid- longitudinal yellowish lines decorated yellow 

patches, laterally with some weavy yellowish lines. Five pairs 

of sigilla arrange mid-dorsally. Ventrum provided with a broad 

longitudinal olive-brown patch between epigastric furrow and 

spinnerets.Anterior spinneret 0.64. Epigynum heavily 

sclerotised, having a transverse concave groove with distinct 

anterior ridge.Male not found. 

Diagnosis : Cephalic region convex, more elevated than 

thoracic region and generally armed posteriorly with two 

tubercles; labium longer than wide, maxillae elongated. Ocular 

quadrangle nearly square a slightly wider behind. Legs very 

long and strong, clothed with spines, metatarsi longer than 

tibiae and patella together. Epigynum sclerotised. 

Materials examined : 1@&, 2.XI.2009, Ukhrul district. 

Coll. Bhubaneshwari, Noren 

2@&, 25.XI.2010, Moreh, Chandel district. 

Coll. Bhubaneshwari, Manoj 

1@& , 4.XII.2010, koubru Leikha, Senapati district. 

Coll.Manoj Bhubaneshwari, Binarani 

1@&, 23.XI.2010, Leimaram, Waroiching, Bishenpur 

district. 

Coll. Kanan, Bhubaneshwari, Manoj 

Distribution : INDIA : Manipur (Dist. Ukhrul, Senapati, 

Chandel, Bishenpur); Tamil Nadu, Karnataka, Uttar Pradesh, 

Madhya Pradesh, Assam, Sikkim, West Bengal, 

Maharashtra,Gujarat, Andaman & Nicobar Island. Myanmar, 

Shri Lanka, China, Australia, Malaysia, Japan, New Guinea. 

Habitat : Commonly found in deep forests and construct 

large webs between the branches of large tall trees on the 

bushes. Commonly found in deep forest and construct large 

webs between the branches of tall trees on the bushes. 

Nephila clavata L. Koch [Fig. 1 & 2] 

Nephila clavata Koch, L., 1878, Ver. Zool.-bot. Ges. 

Wien. 27:741. 

Description female from Ukhrul and male from 

Waroiching. 

Female 

Total length – 18.10; Cephalothorax – 5.40 long, 3.30 

wide; Abdomen – 14.35 long, 7.20 wide 

Colour in alcohol : Carapace brown chelicerae reddishbrown, 

labium dark brown with pale distal margins. Sternum 

dark brown with yellowish patch. Legs black, femur and tibia 

with yellowish bands. Abdomen dorsum yellowish brown, 

ventrum olive brown with some yellowish patches. 

Cephalothorax : Longer than wide slightly narrower in 

front, cephalic region provided with v-shaped yellowish patch 

posteriorly and lighter in colour than thoracic region; dark 

row with shallow fovea. Eyes : Eight eyes in two rows, slightly 

narrower in front than behind, median eyes equal in size, lateral 

eyes sub-equal in size, Closer than median eyes and situated 

on prominient tubercles. Maxillae : 1.86 long, 1.13 wide, broader 

distally, provided with distinct scapulae Labium : 1.23 long, 

1.13 wide, dark brown with pale distal margin. Chelicera : 3.76 

long, strong, stout, reddish brown with 3 promargin and 4 

retromargin teeth on cheliceral furrow. 

Sternum : 2.65 long, 2.36 wide, triangular, a dark-brown 

with a J- shaped, yellowish patch clothed with pubescence 

and hairs. Legs : Legs very long, strong, black but femora 

brown ; femora & tibiae with distinct yellowish bands in middle 

and clothed with hairs & spines. Leg formula 1243 

Spination : 

Leg I : Fe- p=4, r=1, ti- p=7, r=4, met- p=9, r=5, 

Leg II : Fe- p=1, r=1, ti- p=6, r=2, 

Leg III : pat- r=1, ti- p=2, r=2, met- p=4, r=3; Leg 

IV : pat- r=1, ti- p=2, r=1, met- p=2, r=1. 

Palp – Trichobothria are arranged laterally. 

Measurements of legs segments : 


I 12.35 2.30 10.15 14.85 3.05 42.70 

II 10.50 1.80 7.35 11.70 2.60 33.95 

III 5.75 1.20 2.80 4.85 1.85 16.45 

IV 10.20 1.45 5.75 9.80 2.20 29.40 


Pedipalp 1.84 0.50 1.14 - 2.01 5.49 

Abdomen : Abdomen oval, longer than cephalothorax, 

dorsum-yellow in colour, decorated with 4 dark green bands 

arranged horizontally. A dark mid band extending from the 

anterior end of the abdomen upto the posterior end 

intersecting the horizontal bands, 5 pairs of sigilla arranged 

mid-longitudinally. Ventrum olive brown with some yellowish 

patches and reddish brown posteriorly. Spinneret beyond the 

apex of the abdomen, anterior spinnerets length 0.63. Epigynal 

plate small with indistinct curved-ridge. 

Descrpition of male from Waroiching : 

Total length - 7.4; Cephalothorax - 2.79 long, 2.15 wide; 

Abdomen – 4.61 long, 1.39 wide. 

Cephalothorax : Cephalothorax light yellow, laterally dark 

brown, ocular quad 0.82 long; 2.38 wide. PME light yellow in


Fig. 1

Kananbala et. al., : Study on two edible spiders of the Genus: Nephila (Fam. Nephilidae Simon 1894) of Manipur, India 157 

colour, PLE and AME dark in colour, trichobothria present 

just above each of the PLE, anterior row of eyes strongly 

recurved, posterior slightly recurved. Maxilla- 2.11 long, 1.42 

wide; light yellow directed towards each other with brown 

lateral sides covered with small hairs. Labium : 1.41 long, 1.31 

wide; light yellow broad at the middle with few hairs at the 

base. Chelicera : Light yellow, fang dark brown, 0.85 long 

with each 3 retomargin and promargin teeth. Sternum : Slightly 

heart-shaped, 1.42 long, 0.83 wide. Legs : Yellow colour upto 

patella, red and dark band extending upto tarsus with 3 claws. 

Leg formula 1243. Pedipalp : Light yellow upto patella, tibia 

dark brown in colour, trichobothria present in the femur and 

patella, embolus long coiled, papal organ compex type with 

dark colour. 

Spination : 

Leg I : Fe- p=5, r=1, pat- r=1, d=1, ti- p=3, r=4, met- r=1. 

Leg II : Fe- r=2, pat-p=2, r=2, ti- p=2,r=2, met- p=2,r=2. Leg III 

: a==2, r=2, pat- r=1, ti- p=2, r=1, met- p=1, r=1. Leg IV : Fe- p=3, 

r=1, pat- a=1, ti- p=3, r=2, met- p=3, r=1. 


I 7.61 1.55 6.55 9.85 2.66 28.22 

II 6.85 1.50 4.75 7.88 2.23 23.21 

III 3.56 0.69 1.51 2.56 1.34 9.66 

IV 6.19 0.89 3.36 5.76 1.59 17.79 


Pedipalp 0.83 0.37 0.35 - 0.82 2.37 

Measurements of legs segments : 

Abdomen : Dorsum yellow extending from anterior to 

posterior decorated with dark brown mid-dorsal band. Ventrum 

with dark brown, mid-ventral band with shining yellow colour 

ventro-laterally. 

Materials examined : 3@&, 23.XI.2009, Ukhrul District. 

Coll. Bbubaneshwari, Noren 

1B& & 3@&, 23.XI.2010, Waroiching, Bishenpur District. 

Coll. Bhubaneshwari, Manoj, Binarani 

2@&, 25.XI.2010, Chandel District (Moreh) 

Coll. Bhubaneshwari, Manoj, Binarani 

Distribution : INDIA : Manipur ( Ukhrul District, 

Bishenpur District, Chandel District), Sikkim, West Bengal, 

Meghalaya, Andaman & Nicober Island, Laccadive Island; 

Bhutan; Myanmar; Thailand; Pakistan; Japan; China; 

Formosa. 

Habitat : Commonly found in the branches of trees in 

the forest area. 

Diagnosis : Cephalic region convex, and elevated 

without tubercles provided with v-shaped yellow patch. 

Abdomen yellowish in colour, longer than wide. 


The authors thanks to UGC for the financial assistance 

and thanks to the principal, D.M. College of Science, Imphal 

for providing the laboratory facilities. 


Sebastian, P.A and K. V. Peter 2009. Spiders of India. 

Satpathi, C. R. 2004. Predacious spiders of crop pests. 



Effect of Coagulants on Nutrient and Antinutrient Parameters of Soy Tofu 

M.K.TRIPATHI, S.MANGARAJ 

Agro Produce Processing Division, Centra Institute of Agricultural Engineering, Nabi Bagh, Baresia Road, 

Bhopal, M.P. India 

e-mail: tripathimanoj007@gmail.com 

ABSTRACT 

Soymilk has always been a rich source of protein which is 

inexpensive and abundantly available. Tofu is one of the most 

popular soy-products and is prepared by coagulating soymilk. 

The effects of four coagulating agents (CaCl 2 

, MgCl 2 

, CaSO 4 

and MgSO 4 

), on the yield, nutrient, and anti-nutrient 

composition of tofu samples produced was studied. The 

percentage yield for tofu coagulated with CaSO 4 

was 

significantly different (p

TRIPATHI & MANGARAJ : Effect of Coagulants on Nutrient and Antinutrient Parameters of Soy Tofu 159 


The Tofu yield was found to affected by nature of 

coagulants used. The % yield was modulated by the 

coagulants is presented in Fig. 2. It varied from 64% (for Tofu 

coagulated with CaSO4) to 69 % (for Tofu coagulated with 

MgCl2). Highest yield was found with coagulant B which is 

higher than any other coagulants at used concentration and 

temperature and similar result was also reported by Oboh, 

2006. The processing conditions and soybean varieties used 

in tofu preparation might have affect on Tofu Yield. 

Fig.1. Detail steps for coagulation of soymilk Fig. 2 Yield of Tofu processed with coagulants (A, B, C and D) 

Chemical Analysis : 

Tofu samples were analyzed in triplicates for moisture, 

protein, lipid, fiber and ash using standard methods of analysis. 

The chemical composition was estimated according to AOAC 

(AOAC 2005): Moisture (AOAC, 967.08); Protein by Kjeldahl 

(AOAC, 988.05); Fat by Soxhlet (AOAC, 2003.06); Fiber 

(AOAC, 958.06) and Ashes (AOAC, 942.05). Carbohydrate 

was estimated by difference. Trypsin inhibitory activity was 

determined by the method of Kakade, et al., 1969. 

Determination of textural properties of Tofu : 

Texture Profile Analysis uses mechanical parameters of 

texture, which imitate the action of jaws, and the texture 

analyser is programmed to compress a bite-size piece twice in 

a reciprocating motion. Samples of 2x2 cm dia. cylinders of 

tofu, cut from the main block with a cork borer, then trimmed 

to length by cutting with fine wires set in a frame 2 cm apart 

were used for texture measurement. A two-cycle compression 

test was used, in which the probe touched the sample and 

compressed it to 75% of the brick height (15 mm) at a crosshead 

speed 18 mm/min, returned and repeated the test using 

the same parameters and recorded the fracturability, hardness, 

cohesiveness, gumminess, springiness, chewiness and 

resilience as forces. 

Total bacterial count (TBC) : 

Plate count agar was used for the determination of total 

viable counts. All plates were triplicated, incubated at 37 0 C 

for 48 h, and viable cell numbers were determined as colony 

forming units (CFU) per mL. 

Proximate analysis of Tofu : 

The proximate composition of the Tofu prepared by all 

four coagulants is presented in Table 1. The moisture content 

of Tofu samples varied from 72.01 % to 75.18%. The variation 

in the moisture content of Tofu prepared with different 

coagulants is probably due to the differences in gel network 

within the Tofu particles that is influenced by different anions 

and its ionic strengths towards the water holding capacity of 

soy protein gels. It may also be due to the unique coagulating 

properties of the coagulants used. Tofu is relatively high in 

protein, about 10.7% for firm Tofu and 5.3% for soft silken tofu 

with about 5% and 2% fat respectively as a percentage of 

weight. The protein content Tofu sample in experiments was 

found 8.53 % to 10.5% of weight (Table 1). 

Table 1. Proximate composition of Tofu (% dry weight) 

Coagul 

ants 

Concentr 

ation 

(mM) 

Temperature 

(°c) 

Moisture* Fat* Ash* 

(%) (%) (%) 

Protein* Carbohy 

(%) drate* 

(%) 

CaCl2 50 mM 90 0 C 73.93 6.95 2.87 10.5 5.75 

MgCl2 50 mM 90 0 C 73.45 7.51 4.32 9.28 5.44 

Ca SO4 50 mM 90 0 C 75.18 5.82 3.92 9.75 5.33 

MgSO4 50 mM 90 0 C 72.01 7.58 4.54 8.53 7.34 

*Average of 3 replicates 

Textural characteristics of Tofu : 

Hardness and gumminess were shown to be the most 

significant indicators of Tofu texture. For tested chemical 

coagulants, significant correlations were found between Tofu 

hardness and volume of separated whey. No correlation 

between the pH of Tofu whey and any of the studied texture


characteristics of Tofu was observed indicaticating that the 

extent of soymilk gelation and tofu texture is not determined 

by a single characteristic but rather results from a combination 

of factors. The hardness of CaCl 2 

-Tofu had on an average the 

highest value compared to other tested coagulants. The results 

also confirmed the well-known fact that the use of a suitable 

concentration of the quick-acting coagulants is more critical 

than that of the slow-acting coagulants in Tofu making. 

Utilization of each coagulant in Tofu making influenced the 

tofu texture resulted, particularly in firmness of Tofu. 

Antinutrient (Trypsin Inhibitors) characteristics of Tofu : 

Trypsin inhibitor is an anti-nutritional factor that affects 

the protein digestibility (Liener, et al., 1980). Though it is heatlabile, 

the heat treatment insolubilizes the much-valued 

proteins and, more importantly, excessive heat treatment can 

cause loss of amino acids in soy proteins. There are very 

limited data on the effect of coagulants on the level of trypsin 

inhibitors in tofu. The trypsin inhibitor levels in Tofu found to 

ranges from 1.46 TIU/mg proteins in Tofu coagulated with 

MgSO 4 

to 1.94 TIU/mg proteins in Tofu coagulated with CaCl 2 

(Fig.3). 

Fig. 3. 

Anti-nutrient (Trypsin Inhibitor) composition of tofu 

with coagulants (A, B, C and D) 

Microbial quality evaluation of Tofu : 

Table 2 shows the changes of the viable microbial counts 

of tofu prepared with above salt coagulants during storage at 

10 0 C for 8 days. All Tofu had initial bacterial concentrations of 

10 2 CFU/g at 0 day of storage. The viable microbial counts of 

tofu prepared with all coagulants increased more rapidly with 

time at 10 0 C. Kim and Lee 1992 reported that Tofu spoilage 

would start when viable counts were above 10 7 CFU/mL. Based 

on this report, the shelf life of Tofu was found to 2 to 3 days. 

Tofu is one of the most popular soy-products and is 

prepared by coagulating soymilk. The quality of Tofu depends 

on several parameters such as coagulation method, processing 

condition, texture, the content of two storage protein 

Table 2. Microbial quality evaluation of tofu 

Days 

Total bacterial counts (cfu/ml) 

0 1.8X10 2 

2 1.2X10 4 

4 6X10 8 

6 3X10 9 

8 9X10 9 

components glycinin and -conglycinin and their ratio. The 

yield, nutrient and anti-nutrient contents of tofu are greatly 

affected by the type of coagulant used. The results showed 

that the concentration and type of coagulant had a great 

influence on the properties of the Tofu. All coagulants used in 

investigation were able to coagulate the soymilk at selected 

concentrations of 50mM. Depending on the type of coagulant 

used, as well as stirring during coagulation and pressure 

applied to the curd, Tofu ranges in hardness from soft to firm 

with a moisture content of 72 to 75% and protein content of 8 

to 10% dry weight basis. Tofu made with CaCl 2 

and MgCl 2 

was 

coarse, granular, and hard, whereas calcium sulfate gave a 

smooth and soft texture. 


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Maryland, USA. 

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of bench and production scale methods for making soymilk and 

tofu from 13 soybean varieties Food Res. Int. 30(9): 659–668. 

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Applications, Garant Inc, pp. 200-214. 

Food and Drug Administration (FDA). 1999. http://www.fda.gov/food/ 

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cientifica greementssa/ucm074740.htm. 

Kakade, M.L. Simons, S.N. Liener, I.E.1969.An evaluation of 

naturalvs.synthetic substances for measuring the antitryptic activity 

of soybean samples. Cer. Chem. 46: 518-526. 

Liener, I.E. Kakade, M.L. 1980. Toxic constituents of plant food stuffs 

In: I.E. Liener, 2nd ed. Academic Press: New York pp. 7–72. 

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K. Liu (ed.) (International Thomson Publishing, New York: pp. 

442-477. 

Messina, M. 1999. Legumes and soybeans: overview of their nutritional 

profiles and health effects. Am. J. Clin. Nutr., 70: 439S-450S. 

Oboh, G. 2006. Coagulants modulate the hypocholesterolemic effect 

of tofu (coagulated soymilk). Afr. J. Biotech., 5(3): 290–294. 

Shokunbi, O.S. Babajide, O.O. Otaigbe, D. O. Shobowale, O.A. Ajiboye, 

A.A. Tayo, G.O. 2011. Coagulants Modulate the Yield and 

Micronutrient Composition of Tofu. WJDFS, 6(1): 67-70. 



Integrated Weed Management Studies on Weed Flora and Yield in Kharif Maize 

BIRENDRA KUMAR 1 RANVIR KUMAR 2 SUMAN KALYANI 2 AND MIZZANUL HAQUE 1 

1 

Deptt. of Agronomy, Bihar Agricultural College, Sabour, Bhagalpur, Bihar 813 210 

2 

B.P.S. Agricultural College, Purnea City, Purnea, Bihar 854 302 

e-mail: ranvir.bausabour@gmail.com 2 

ABSTRACT 

A field experiment was conducted during the Kharif season of 

2011-12 at Bihar Agricultural University, Sabour, ( Bihar) to 

evaluate the effectiveness and economically integrated weed 

management method in maize (Zea mays L.) .Weed management 

had a positive influence on growth, yield attributes and yield of 

the maize. Manual weeding at 15 and 30 days after sowing 

(DAS) with in Zero tillage ( ZT) - Glyphosate Pre Plant followed 

by Atrazine+ Halosulfuran (1.0 kg + 90 g a.i./ha) as Post 

emergence, Conventional tillage( CT) - Atrazine + 

Halosulfuran @ (1.5kg+90 g a.i/ha) as Post emergence and 

Zero tillage (ZT) - Glyphosate Pre plant followed by 

Topramezone+Atrazine @( 40 ml + 500 g a.i/ha) as Post 

emergence proved equally effective in increasing most of the 

growth parameters, yield attributes, yield and economic 

advantage. The effect due to different weed management 

practices on grain yield of maize was found to be statistically 

significant. The maximum mean grain yield of (68.2 q/ha) was 

recorded from the plots where two hand weeding at 15 and 30 

DAS was performed and was statistically at par with the mean 

grain yield obtained under different weed management 

practices i.e. ZT - Glyphosate Pre Plant followed by Atrazine+ 

Halosulfuran (1.0 kg + 90 g a.i./ha) as POE (67.1 q/ha), CT- 

Atrazine + Halosulfuran @ (1.5kg+90 g a.i/ha) as POE (64.1 q/ 

ha), ZT-Glyphosate Pre plant followed by 

Topramezone+Atrazine @( 40 ml + 500 g a.i/ha) as POE (63.6 

q/ha), and the grain yield obtained these were significantly 

superior to the grain yield obtained under rest of the weed 

management practices. Yield advantages due to different weed 

management practices over weedy check were mainly attributed 

due to enhance yield attributing parameters as a result of lower 

weed population, biomass along with higher weed control 

efficiency. The highest net return Rs, 34856/ha and maximum 

benefit: cost ratio of 1.2 was noted in treatment ZT-Glyphosate 

Pre plant followed by Topramezone+Atra zine @( 40 ml + 500 

g a.i/ha) POE and lower value of net return Rs, 22980/ha and 

benefit: cost ratio of (0.82) were recorded under weedy check. 

Key words Conventional tillage, Economics, Maize, Weed density, 

Yield, Zero tillage 

In Bihar, total area under Maize (Zea mays L.) is 6.98 

lakhs ha, producing 21.11 matric tons and with average 

productivity of 3025 kg/ha (Anon., 2012). The area under maize 

is increasing in Bihar due to favorable prices and increasing 

in demand. Tillage is a critical practice in crop production as it 

provides favourable condition for crop growth and 

development. It is reported that the normal tillage may not be 

required for getting optimum crop yield. Rainy season maize 

suffers from severe weed competition and depending upon 

the intensity, nature, stages and duration of weed infestation 

causes yield losses varying from 28-100% (Patel, et. al., 2006). 

A wide spaced crop suffers from heavy weed infestation due 

to slow initial growth particularly under Kharif season. Weed 

infestation is one of the major constraints for low yield of 

maize as weeds compete with crop plants for essentials inputs. 

Weed depletes 30-40% of applied nutrients from the soil. The 

losses caused by weeds exceed the losses from any other 

category of agricultural pests was noticed by Padhi and 

Panigrahi, 2006 and Sharma and Behera, 2009. Under such a 

situation, the concept of zero tillage offers ample scope for 

combating weeds without any threat to eco-system. 

To realize the maximum benefit of applied costly inputs 

and high yields, control of weeds is inevitable. Growing 

intercrops in widely spaced maize crop not only reduce 

intensity of weeds but also gives additional yield was noticed 

by Hussain Nazim, et. al., 2003. Hence, keeping the above fact 

in mind, the present investigation was carried out to assess 

the possibility of increasing crop production per unit area by 

introducing effective tillage and weed control method in rainy 

season maize. 


A field experiment was carried out during rainy season 

of 2011-12 at Bihar Agricultural University, Sabour, Bihar 

(25 O 04’ N Latitude, 87 O 04’ E Longitude and 37.19 meter above 

mean sea levels), in a randomized block design with three 

replications. In conventional tilled treatment three ploughing 

followed by planking was done. Under zero tilled condition, 

crop was sown directly and glyphosate @ 1.0 kg a.i/ha was 

sprayed one week before sowing of the crop to kill the existing 

weeds. The treatments comprised 10 weed management 

practices i.e., weed control treatments were : Un-weeded check, 

CT- HW at 15 and 30 DAS,CT- Maize+ black gram as intercrop, 

CT- Atrazine@1.5kg a.i/ha as Pre-em., CT- Atrazine + 

Halosulfuran @ (1.5kg+90 g a.i/ha) as Post emergence, CT- 

Halosulfuran @ 90 g a.i/ha as POE, CT- Topramezone+Atrazine 

@( 40 ml + 500 g a.i/ha) as POE, ZT-Glyphosate Pre plant 

followed by Atrazine +Halosulfuran(1.0 kg + 90 g a.i/ha) as 

POE, ZT-Glyphosate Pre plant followed by Topramezone + 

Atrazine @ (40 ml + 500 g a.i/ha) as POE, ZT-Glyphosate Pre 

plant followed by Maize+ black gram as intercrop ‘DHM 117’


maize was sown on 01 July in 2011-12. In maize, 1/3 N was 

applied basal along with P and K and the remaining nitrogen 

were applied in two splits only in rows of maize each at knee 

high and pre-taselling stage. Pre-emergence and Postemergence 

herbicides were applied at next day and 30 days 

after sowing using water volume of 800 liters/ha. The data on 

weed population, weed density and weed control efficiency 

were recorded at different stages of crop. The experimental 

soil was sandy-loam in texture with pH 7.2. The organic carbon, 

electrical conductivity and available nitrogen, phosphorus 

and potash were 0.58%, 0.107ds/m, 270.6, 15.36 and 290.15 kg/ 

ha, respectively. The rainfall received during the crop season 

of respective years was 1202 mm. Cost of cultivation and gross 

return were calculated on the basis of prevailing market prices 

of different inputs and produces, respectively. 


Weed growth, Density and weed dry weight 

Dominant weed species present in the experimental site 

were Cynodon dactylon L, Cyperus rotundus L., Echinochloa 

colonum L, Echinochloa crusgalli L, Eleusine indica L, 

Phyllanthus niruri L, Euphorbia hirta L , Amaranthus viridis 

L,Commelina benghalensis L and Parthenium hysterophorus L. 

Weed density was significantly affected due to different 

tillage systems. There was reduction in total weed density in 

ZT when compared with CT. Higher weed density in CT may 

be due to better tilth and exposure of weed seeds to the upper 

soil layers (Singh, et. al., 2001). Density of all grasses was 

maximum in CT system. All the herbicides reduced weed 

density and weed dry weight over weedy check. Minimum 

population of all major weed species were significantly reduce 

due to two hand weedings at 15 and 30 DAS. 

Growth and yield attributes of maize 

Different weed management practices significantly 

influenced the growth, yield attributes and yield of maize crop 

(Table1) .Two hand weeding at 15 and 30 days after sowing 

being statistically at par with the weed control treatment, 

ZT-Glyphosate Pre Plant followed by Atrazine+ Halosulfuran 

(1.0 kg + 90 g a.i./ha) as POE recorded significantly higher 

values of growth, yield attributes and yield to the rest of the 

weed control treatments. This might be probably due to the 

creation of modified micro-climate inturns of physical 

environment for mechanical manipulation of soil and lower 

crop –weed competition under two hand weeding might have 

led to better yield components and thus resulted in higher 

yield (Mundra, et. al., 2003).during hand weeding and being 

persistence and broad spectrum control of weeds keep the 

population of weed under check by arresting or inhibiting the 

germination of weed seeds and arresting the growth and 

development of weeds which provide weed-free environment 

to the crop resulted into better manifestation of growth and 

yield attributes and ultimately enhanced the crop yield. The 

results are in conformity with those reported by Singh, et. al., 

2005. Yield advantage due to different weed management 

practices over weedy check were mainly attributed for better 

yield attributing parameters and cooperatively less weed 

population weed biomass along with higher weed control 

efficiency. Interaction effect of tillage and weed control method 

on grain yield was found significant. 

Total productivity 

Different weed management practices significantly 

influenced the maize-equivalent yield. Hand weeding at 15 

and 30 DAS being statistically at par with ZT-Glyphosate Pre 

Plant followed by Atrazine+ Halosulfuran (1.0 kg + 90 g a.i./ 

ha) as POE recorded significantly higher grain yield to the 

rest of the weed control treatments. 

Table 1. Effect of weed management on growth, yield attributes and yield of maize. 

Treatment 

Plant height Weight of one 100 grains Mean Grain Yield 

(cm) cob(gm) weight (g) ( q/ha) &MEY 

W1-Un-weeded check 140.4 210 30 51.0 

W2- CT-HW at 15 & 30 DAS 157.8 216 36 68.2 

W3- CT-Maize+ black gram as intercrop. 145.6 212 33 56.5 (65.5) 

W4- CT-Atrazine@1.5kg a.i/ha as Pre-em 148.3 213 34 61.4 

W5- CT-Atra zine@1.5kg a.i followed by Halosulfuron @ 90 g a.i/ha as POE 150.6 214 35 64.1 

W6- CT-Halosulfuron @ 90 g a.i/ha as POE 149.5 213 34 62.6 

W7- CT- Topramezone+Atrazine @ ( 40 ml + 500 g a.i/ha) as POE 149.8 213 34 62.7 

W 

8- ZT-Glyphosate Pre plant followed by Atrazine +Halosulfuron(1.0 kg+90 g a.i/ha) 

as POE 

W 

9- ZT-Glyphosate Pre plant followed by Topramezone+Atra zine @( 40 ml + 500 g 

a.i/ha) POE 

156.1 215 35 67.1 

150.3 214 34 63.6 

W10- ZT-Glyphosate Pre plant followed by Maize+ black gram as intercrop 148.4 212 34 57.7 (66.4) 

SEm± 0.84 0.21 0.93 3.9 

CD (P=0.05) 1.8 0.45 1.93 8.3

KUMAR et. al., : Integrated Weed Management Studies on Weed Flora and Yield in Kharif Maize 163 

Intercropping systems significantly reduced the weed 

population and weed dry weight than sole cropping (Table 

2). Intercropping with maize + black gram was the more 

effective in suppressing weeds and recorded the less weed 

population and weed dry weight. The reduction in weed 

population and weed dry biomass in intercropping systems 

may be attributed to shading effect and competition stress 

created by the canopy of more number of crop plants in an 

unit area having suppressive effect on associated weeds, thus 

preventing the weeds to attain full growth. Intercropping 

system of maize + black gram exhibited land equivalent ratio 

greater than sole cropping, indicating greater biological 

efficiency of intercropping system and thereby resulting in 

higher productivity per unit of space. Planting of maize and 

black gram recorded the higher land equivalent ratio. All the 

weed control treatments significantly reduced the density and 

dry weight of weeds compared with weedy check. Hand 

weeding at 15 and 30 DAS proved most effective in reducing 

the population of weeds and weed dry matter production. 

The performance of hand weeding, ZT-Glyphosate Pre Plant 

followed by Atrazine+ Halosulfuran (1.0 kg + 90 g a.i./ha) as 

POE, CT- Atrazine + Halosulfuran @ (1.5kg+90 g a.i/ha) as 

POE and ZT-Glyphosate Pre plant followed by 

Topramezone+Atrazine @( 40 ml + 500 g a.i/ha) as POE was 

Table 2. Effect of weed management on growth, yield attributes and yield of maize. 

Treatment 

W 

1-Un-weeded check 

Cob length Cob diameter Number of grain Number of 

(cm) (cm) row per cob grain per row 

12.4 4.3 13 35 

W 

2- CT-HW at 15 & 30 DAS 

17.0 5.3 15 42 

W 

3- CT-Maize+ black gram as intercrop. 

13.6 4.4 13 39 

W 

4- CT-Atrazine@1.5kg a.i/ha as Pre-em 

14.2 4.5 14 40 

W 

5- CT-Atra zine@1.5kg a.i followed by Halosulfuron @ 90 g a.i/ha as POE 

16.1 4.8 14 41 

W 

6- CT-Halosulfuron @ 90 g a.i/ha as POE 

15.0 4.6 14 40 

W 

7- CT- Topramezone+Atrazine @ ( 40 ml + 500 g a.i/ha) as POE 

15.0 4.6 14 40 

W 

8- ZT-Glyphosate Pre plant followed by Atrazine +Halosulfuron(1.0 kg+90 g a.i/ha) as 

16.5 5.1 15 41 

POE 

W 

9- ZT-Glyphosate Pre plant followed by Topramezone+Atra zine @( 40 ml + 500 g 

15.6 4.7 14 40 

a.i/ha) POE 

W 

10- ZT-Glyphosate Pre plant followed by Maize+ black gram as intercrop 

13.7 4.5 14 39 

SEm± 0.21 0.17 0.61 0.78 

CD (P=0.05) 0.45 0.36 1.2 1.65 

Table 3. Effect of weed management on weed biomass ,weed density, weed control efficiency . 

Treatment 

Weed population 

(No/m2) at 30 

DAS 

Weed 

population 

(No/m2) at 

60 DAS 

Dry wt of weed 

(gm/m2)at 30 

DAS 

Dry wt of weed 

(gm/m2) at 

60 DAS 

Weed 

control 

efficiency at 

30 DAS 

Weed 

control 

efficiency at 

60 DAS 

235 238 71.97 81.27 - - 

W 

1-Un-weeded check 

W 

2- CT-HW at 15 & 30 DAS 

57 67 24.00 27.03 66.5 66.7 

W 

3- CT-Maize+ black gram as intercrop. 

92 115 40.00 44.53 44.4 45.2 

W 

4- CT-Atrazine@1.5kg a.i/ha as Pre-em 

72 80 28.00 32.07 61.0 60.5 

W 

5- CT-Atra zine@1.5kg a.i followed by Halosulfuron @ 

60 72 25.43 29.10 64.6 64.1 

90 g a.i/ha as POE 

W 

6- CT-Halosulfuron @ 90 g a.i/ha as POE 

76 86 28.30 31.50 60.6 61.2 

W 

7- CT- Topramezone+Atrazine @ ( 40 ml + 500 g 

68 77 26.07 30.33 63.7 62.6 

a.i/ha) as POE 

W 

8- ZT-Glyphosate Pre plant followed by Atrazine 

59 71 25.00 28.57 65.2 64.8 

+Halosulfuron(1.0 kg+90 g a.i/ha) as POE 

W 

9- ZT-Glyphosate Pre plant followed by 

62 82 26.80 29.50 62.7 63.7 

Topramezone+Atra zine @( 40 ml + 500 g a.i/ha) POE 

W 

10- ZT-Glyphosate Pre plant followed by Maize+ black 

90 110 28.30 31.93 60.6 60.7 

gram as intercrop 

SEm± 8.4 6.7 1.9 2.3 - - 

CD (P=0.05) 17.8 14.2 4.0 4.8 - -


Table 4. 

Effect of weed management on yield of maize, maize equivalent yield, cost of cultivation, gross return, net return, 

benefit : cost ratio. 

Treatment 

Mean grain &maize General cost of cultivation + Cost 

equivalent yield (t/ha) due to herbicides (Rs./ha) 

W1-Un-weeded check 51 28020 

Gross return 

(Rs./ha) 

Net return 

(Rs./ha) 

Benefit : cost 

ratio (Rs.) 

W2- CT-HW at 15 & 30 DAS 68.2 28020+8640= 36,660 68200 31540 0.86 

W3- CT-Maize+ black gram as intercrop. 56.5(65.5) 28020+1632=29652 56500(65500) 26848(35848) 0.90(1.20) 

W4- CT-Atrazine@1.5kg a.i/ha as Pre-em. 61.4 28020+1392=29412 61400 31988 1.08 

W 

5- CT-Atra zine@1.5kg a.i followed by 

Halosulfuron @ 90 g a.i/ha as POE . 

64.1 28020+1692=29712 64100 34388 1.15 

W6- CT- Halosulfuron @ 90 g a.i/ha as POE . 62.6 28020+300=28320 62600 34280 1.21 

W 

7- CT- Topramezone+Atrazine @ ( 40 ml + 

500 g a.i/ha) as POE . 

62.7 28020+4724=32744 62700 29956 0.91 

W 


Atrazine +Halosulfuron(1.0 kg+90 g a.i/ha) as 67.1 24020+9213=33233 67100 33867 1.01 

POE 

W 


Topramezone+Atra zine @( 40 ml + 500 g 

63.6 24020+4724=28744 63600 34856 1.21 

a.i/ha) POE 

W 


Maize+ black gram as intercrop 

57.7(66.4) 24020+1632=25652 57700(66400) 32048(40748) 1.24(1.58) 

SEm± 3.9 688.5 3256.6 1121.5 0.091 

CD (P=0.05) 8.2 2162 9896 3398 0.29 

statistically alike and inturn were significantly superior to the 

remaining weed control treatments (Table 2). 

Significant reduction of weed density and weed dry 

biomass under hand weeding and mixed herbicides Atrazine+ 

Halosulfuran, Topramezone+Atrazine might be due to the fact 

that these weed control treatments gave almost season-long 

control of weeds obviously due to their persistence in soil for 

a sufficiently long time and broad spectrum control of weeds. 

The results are in conformity with those reported by Ram, et. 

al., 2003. All the weed control methods resulted significant 

increase in grain and biological yield over weedy check. 

Economics 

Atra zine @( 40 ml + 500 g a.i/ha) POE had the highest 

net return(Rs,34856/ha) and B:C ratio (1.21) as compared to 

rest of the weed control methods. 

It may be concluded that Zero tillage and two hand 

weeding at 15 & 30 days after sowing appeared to be the best 

in reducing weed growth and producing maximum grain yield 

in maize. 


Hussain, Nazim, Imran Haider, Shamsi, Khan Sherin, Habib Akbar and 

Wajid Ali, Shah: 2003. Effect of legume intercrops and nitrogen 

levels on the yield performance of maize. Asian Journal of Plant 

Science 2(2): 242–46. 

Mundra, S.L., Vyas, A.K and Maliwal, P.L. 2003. Effect of weed and 

nutrient management on weed growth and productivity of maize 

(Zea mays L.). Indian Journal of weed science 35 (1 & 2):57-61. 

Padhi, A.K. and Panigrahi, R.K. 2006. Effect of intercrop and crop 

geometry on productivity, economics, energetic and soil fertility 

status of maize based intercropping systems. Indian Journal of 

Agronomy 51(3):65-67. 

Patel, V.J., Upadhyay, P.N., Patel, J.B and Meisuriya, M.I. 2006. Effect 

of herbicide mixture on weeds in Kharif maize(Zea mays L.) under 

middle Gujarat conditions. Indian Journal of Weed science 38(1 & 

2): 54-57. 

Ram, B., Choudhary, A.S. Jat, A.S. and Jat, M.L. 2003. Effect of 

integrated weed management and intercropping systems on growth 

and yield of pearlmillet (Pennisetum glaucum). Indian Journal of 

Agronomy 48(4): 254–56. 

Sharma, A.R. and Behera, U.K. 2009. Recycling of legume residues for 

nitrogen economy and higher productivity in maize-wheat cropping 

system. Nutrition Cycling Agroecosystem, 83: 197–10. 

Singh, Mahender, Singh, Pushpendra and Nepalia, V. 2005. Integrated 

weed management studies in maize based intercropping system. 

Indian Journal of Weed Science, 37(3 & 4): 205–08. 



Influence of Pseudomonas putida on the Yield of Agaricus bisporus (Lange) Imbach 

PRABHAT KUMAR SINGH, ABHILASHA A. LAL , SOBITA SIMON AND SATISH SHARMA 1 

Department of Plant Protection, SHIATS (Deemed to- be University) Allahabad, U.P., India. 

1 

School of Bio-technology (FOA), Sher-e-Kashmir University of Agriculture Sciences and Technology, 

Kashmir, India. 

e:mail: prabhat9791@gmail.com, aalalplantpathology@gmail.com 

ABSTRACT 

In cultivation of button mushroom [Agaricus bisporus (Lange) 

Imbach], casing layer that is nutritionally deficient to compost 

is believed to trigger the fruit body formation and this is 

conducted by the bacterial community residing in casing layer. 

Therefore, relationship between different concentrations (1x 

10 4 ,1x 10 6 ,1x 10 8 ,1x 10 10 , 1x 10 12 ) of bacterial inoculants 

(Pseudomonas putida) and yield of Agaricus bisporus were 

determined. Available data showed a significant difference in 

yield in respect to concentration of bacterial inoculant added. 

In addition, concentration 1x 10 8 cfu/ml was found to be high 

yielding concentration, which took minimum case run period 

together with higher biological efficiency compared to other. 

Key words White button mushroom, Pseudomonas putida, Bacterial 

inoculants, Yield. 

Agaricus bisporus is the most cultivated species of 

edible mushroom and it is the most popular cultivar among 

the artificially grown fungi of the world that contributes about 

31.8% to the global mushroom cultivation and 85% of the 

total produce in india (Angrish et al.,2003). In commercial 

mushroom cultivation, it is necessary to cover the vegetative 

growth of Agaricus bisporus, which takes place in the compost 

(consisting of composted straw and other plant derived 

materials),with a layer of soil and FYM or various combination 

of similar materials. Among these use of farm yard manure 

(FYM) as a casing medium for mushroom cultivation has been 

vogue in Indian subcontinent because of its easy availability 

and non-availability of peat moss generally used for casing in 

Europe and USA. The application of these materials, termed 

the casing layer, is essential for initiation of basidiomata 

(Eger,1972; Flegg et al., 1985). Bacteria present in casing layer 

considerably influence the growth and morphogenesis of 

Agaricus bisporus production. It supports beneficial microbial 

populations that release growth stimulating substances which 

are reportedly involved in stimulating the initiation of 

pinheads. Several reports are available on the beneficial effects 

of casing soil microbes, especially Pseudomonas putida and 

Alcalgenes faccalis, on Agaricus bisporus (Rainey et al., 1990). 

This work describes the role of population density of 

Pseudomonas putida in the production of Agaricus bisporus. 


Cultivation of Agaricus bisporus : 

Long method of composting (LMC) was adopted using 

the method proposed by Mantel et. al., 1972 using newly 

harvested wheat straw. The formulation given by Singh and 

Mishra, 2006 was modified depending on the availability of 

ingredients locally. 

Spawn was procured from Department of Plant 

Pathology, Chandra Shekher Azad University of Agriculture 

and Technology, Kanpur (U.P). Having completed the 

composting process, thorough spawning was done @75gm/ 

10kg compost. The spawned compost weight 7kg was filled in 

the bags at Mushroom Crop Room, Department of Plant 

Protection, SHIATS, Allahabad. Room temperature, condition 

of spawn run of each bag recorded separately. 

A mixture of FYM and garden soilin the proportion of 

2:1 was used as casing mixture. The mixture was pre- treated 

with 2% formalin two weeks before to eliminate undesirable 

microorganisms. Casing was done at a thickness of 2-3 cm 

and again bags were kept in the dark crop room. The 

temperature and relative humidity were maintained at 14- 18 0 C 

and 80-90% respectively. Temperature during cropping, time 

taken to complete case run, relative humidity in the crop room 

and number of days to pinhead initiation were recorded. 

Inocula preparation : 

The culture of bacterial inoculum (Pseudomonas putida) 

was procured from Department of Plant Pathology, IARI, New 

Delhi, India. 

Inocula were prepared by growing the selective strains 

in King , s B broth medium. After incubation at 30 0 C for 72h, the 

densities of culture were determined. Then cultures were 

diluted further in King , s B broth until final bacterial cell 

numbers were 1x10 4 ,1x10 6 ,1x10 8 ,1x10 10 ,1x10 12 cells/ml . Bacterial 

suspension (77ml/bag) was sprayed in different treatments at 

the time of casing. The harvesting began when buttons were 

fully-grown (but not yet open), and total harvest was recorded 

in each bag.Biologicalefficiency (BE) of the compost was 

calculated using the following formula.



The results of this study showed that there is close 

relation between population density of Pseudomonas putida 

in the casing soil and yield of Agaricus bisporus. 

Concentrations inoculated in casing soil,1x10 8 cfu/ml 

recorded significantly superior over all concentrations, 

required minimum average number of days (13.4) for mycelium 

run in casing soil, highest yield (1.268 kg/bag) and highest 

biological efficiency(27.806%) followed by 1 x 10 10 , 1 x 10 12 , 1 

x 10 6 and 1 x 10 4 cfu/ml as compared to control. Number of 

days for mycelium run increased with the incensement (1 x 

10 10 , 1 x 10 12 cfu/ml) of inocula concentration and also with 

decreasment (1 x 10 6 , 1 x 10 4 cfu/ml) of inocula concentration. 

Table1. 

Effect of bacterial inoculant (Pseudomonas 

putida) on the number of days required for 

mycelium run in the casing, yield of Agaricus 

bisporus and on the biological efficiency of compost 

Concentration Av. No. of days* Yield (kg/bag)* BE (%)* 

Control 17.20 0.316 8.391 

1x10 12 cfu/ml 15.00 0.698 17.868 

1x10 10 cfu/ml 14.60 0.756 19.128 

1x10 8 cfu/ml 13.40 1.268 27.806 

1x10 6 cfu/ml 15.20 0.575 16.478 

1x10 4 cfu/ml 15.60 0.492 13.776 

CD at 5% 0.719 0.184 3.149 

*Average of five replications 

Mushroom yield dramatically increased in concentration 

of 1x10 8 cfu/ml but decreased with increasment of concentration 

(1x10 10 , 1x1012cfu/ml) and also with decreasment of 

concentration (1x10 6 , 1x104cfu/ml). 

Biological efficiency of compost prepared by LMC 

ranged between 13-28%. Highest biological efficiency was 

recorded in bags where concentration inoculated 1x10 8 cfu/ml 

(27.806) followed by 1x10 10 ,1x10 12 ,1x10 6 and 1x10 4 as compared 

to control (Plates 1 and 2). This indicates that the biological 

Fig. 1. Control bag 

Fig. 2. Treated bag with Pseudomonas putida (10 8 cfu/ml) 

efficiency of compost can be improved by inoculation of 

Pseudomonas putida in the casing mixture (Table 1). These 

results are in accordance with the earlier work of Hossein et 

al. (2011), which reported the role of bacterial and cyanobacterial 

culture on growth and yield of Agaricus bisporus. 

In cultivation of Agaricus bisporus, casing soil is major 

element. Population of Pseudomonas in the casing layer on 

which the mushroom fruit body develops is very important. 

We conclude that inoculation of Pseudomonas putida in 

casing soil in 1x10 8 cfu/ml concentration is very efficient for 

increasing yield and biological efficiency (BE) of compost. 


The authors are thankful to Hon ’ ble Vice-Chancellor, 

SHIATS for the facilities provided during the experiment. 


Angrish, M., Sodhi, H.S., Khanna, P.K. and Arora, C. L. 2003. Ideal 

casing material for Agaricus bisporus cultivation under the natural 

climatic condition of Punjab. Mushroom Research, 12: 93-96. 

Eger, G. 1972. Experiments and comments on the action of the bacteria 

on sporophore initiation in Agaricus bisporus.Mushroom Science, 

8: 719-725. 

Flegg,P.B., Spencer, D.M. and Wood, D. A. 1985. The biology and 

technology of cultivated mushroom. John Wiley sons. pp.347. 

Hossein, R. Eskash, A. and Shariatmadari, Z. 2011. Effect of bacterial 

and cyanobacterial culture on growth, quality and yield of Agaricus 

bisporus .International conference on mushroom biology and 

mushroom products. pp. 411- 416. 

Mantel, E.F.K. and Agarwal, P.K. 1972. A guide to mushroom cultivation 

unit, Directorate of Extension. Ministry of Agriculture, New Delhi. 

Rainey, P.B. and Cole, A.L. 1990. A model system for examining 

involvement of nbacteria in basidiome initiation of Agaricus 

bisporus. Mycol. Res., 94: 191-195. 

Singh, R. P. and Mishra, K. K. 2006. Mushroom cultivation, Mushroom 

research and training center, GBPUAT, Pantnagar.pp. 13. 



Effect of Varying NPK Levels and Bio-fertilizers on Growth and Yield of Okra 

[Abelmoschus esculentus (L.) Moench] under Sustainable Condition 

PARAM HANS PRASAD AND ABHISHEK NAIK 

Department of Vegetable Crops, Faculty of Horticulture,Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, 

Nadia 741 252, India 

e-mail: abhishek.hort@gmail.com 

ABSTRACT 

An experiment was conducted at Horticultural Research Station, 

Mondouri, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, 

Nadia in West Bengal during rabi season of 2007-08 with 

objectives to find out the suitable combination of biofertilizers 

[Azotobacter, Azospirillium and Phospho Solubilizing Bacteria 

(PSB)] in presence or absence of FYM (15 t/ha) and two levels 

of graded NPK fertilizers (80-60-50 and 40-30-25 N, P 2 

O 5 

,K 2 

O 

kg/ha) on the growth and fruit yield of okra cv. Arka Anamika 

[Abelmoschus esculentus (L.) Moench] and the availability status 

of N, P and K in soil considering the growth, yield and quality 

parameters data revealed that significantly minimum height 

of plant was recorded by the application of PSB + FYM. In 

respect of plant height at 30 days, plant height at 60 days, 

number of branches, number of leaves, number of fruits, fruit 

length, fruit diameter, fruit yield per plot and fruit yield per ha 

were significantly maximum in the plant receiving 50% 

recommended dose of fertilizer (RDF) + Azotobactor + 

Azospirillum + PSB + FYM with good yield (196.97 q/ha) and 

export fruit quality of Okra. 

Key words Okra, Bio-fertilizer, PSB, NPK, Growth, Yield 

Sustainable crop production requires a judicious 

management of all nutrient sources such as organics, chemical 

fertilizers and biofertilizer available to farmers including precise 

application of need-based irrigation to the crops. The main 

objectives of integrated plant nutrient management (IPNM) 

are to maintain and possibly enhance soil fertility through a 

balanced use of chemical fertilizers combined with organic 

and biological sources to improve efficiency of plant nutrients, 

increase crop productivity and minimizing losses to the 

environment. Nutrient management is one of the prime 

considerations for getting higher yield of any crop. Sustainable 

crop production requires a judicious management of all nutrient 

sources such as organic manure, chemical fertilizers and 

biofertilizer available to farmers including precise application 

of need-based irrigation to the crops. Okra or Bhindi 

[Abelmoschus esculentus (L.) Moench] is one of the most 

important vegetables in New Alluvial Zone of West Bengal. 

Keeping in view the important of the above facts and lack of 

sufficient information, the present experiment was undertaken 

with the following objectives: (i) To find out the suitable 

combination of bio-fertilizer in presence or absence of FYM 

and two levels of graded NPK fertilizers on the growth and 

fruit yield of Lady’s finger (ii) To determine the availability 

status of N, P and K in soil considering the growth, yield and 

quality attributes of crop. 


The experiment was carried out at Horticultural Research 

Station (HRS), Mondouri, Bidhan Chandra Krishi 

Viswavidyalaya, Mohanpur, Nadia in West Bengal. The 

experiment was laid out in Randomized Block Design (RBD) 

having nine different nutritional management with three 

replications. The Treatment details are as follows- 

T 1 

: 

T 2 

: 

T 3 

: 

Recommended dose of fertilizer (RDF) + FYM (15 t/ha). 

Biofertilizer (Azotobacter) + FYM (15 t/ha). 

Biofertilizer (Azospirillum) + FYM (15 t/ha). 

T 4 

: Biofertilizer (Phospho Solubilizing Bacteria) + FYM (15 

t/ha) . 

T 5 

: Biofertilizer (Azotobacter) + FYM (15 t/ha) + RDF (50%). 

T 6 

: Biofertilizer (Azospirillum) + FYM (15 t/ha) + RDF (50%). 

T 7 

: Biofertilizer (Phospho Solubilizing Bacteria) + FYM (15 

t/ha) + RDF (50%). 

T 8 

: 

T 9 

: 

Biofertilizer (Azotobacter + Azospirillum + Phospho 

Solubilizing Bacteria) + FYM (15 t/ha) + RDF (50%). 

Biofertilizer (Azotobacter + Azospirillum + Phospho 

Solubilizing Bacteria) + FYM (15t/ha). 

Before preparation of beds well decomposed farm yard 

manure (FYM) was applied and it was evenly mixed with the 

soil. Okra seed was shown directly at a planting distance of 45 

x 30cm. Irrigation and other intercultural operation were done 

as when required. Observation regarding growth viz., plant 

height cm (30 and 60 DAS), number of branch per plant, 

number of leaves per plant and yield contributing characters 

like number of fruit per plant, fruit length (cm), fruit diameter 

(cm), fruit yield per plot and yield of fruits per hectare (q) were 

recorded and statistically analyzed.



Plant height (cm) 

The data presented in Table 1 revealed significant effect 

on plant height at 30 days after sowing and 60 days after 

sowing under different nutrient management practices. The 

plants of the treatment with Biofertilizer (Azotobacter + 

Azospirillum + Phospho Solubilizing Bacteria) + FYM (15 t/ 

ha) + RDF (50%) were recorded the maximum plant height of 

46.96cm (at 30 DAS) and 72.05cm (at 60DAS). The minimum 

plant height of 41.85cm (at 30DAS) and 51.59cm (at 60DAS) 

were recorded in treatment Biofertilizer (Phospho Solubilizing 

Bacteria) + FYM (15 t/ha). Similarly, in both the studies, 

integrated application of FYM (15t/ha) +RDF (50%) along with 

biofertilizers (Azotobacter, Azospirillum and PSB) 

significantly increased the plant height over control. This is 

probably due to the fact that Integrated Nutrient Management 

(INM) practices might have induced favorable conditions for 

proper growth and development of plants. Application of both 

nitrogen and phosphorus from inorganic fertilizer, accelerate 

the synthesis of chlorophyll and amino acid those are 

involved in the major plant processes. The consequence of 

which might be increased the plant height in okra. Similar 

effect has also been reported by Arora, et al. 1991. Integrated 

application of inorganic fertilizers, organic manures and seed 

treatment with bio-fertilizers might have lead to enhance the 

cell division and cell elongation, resulting in better root 

development, increased the uptake of water and nutrients etc., 

leading to increased in plant height in Okra. Similar result has 

also been recorded by Anburani and Manivannan, 2002. 

Significant improvements in plant height due to combined 

application of both organic and inorganic nutrients have also 

been reported by Barani and Anburani, 2004. 

Number of branches and number of leaves/plant 

Data presented in table 1clearly revealed that different 

treatments have significant effect on number of branches and 

number of leaves / plant. The maximum number of branch 

(3.97) and leaves (41.32) was obtained by the application of 

biofertilizer (Azotobacter+ Azospirillum + PSB) +FYM (15t/ 

ha) +RDF (50%) NPK /ha followed by (3.47 and 34.73) 

respectively with application of RDF+FYM (15t/ha) and 

biofertilizer (Azotobacter) +FYM (15t/ha). Among the different 

treatments, biofertilizer (PSB) + FYM (15t/ha) produced 

minimum number of branch (2.58/plat) and minimum number 

of leaves (26.15/plant) at par (2.89 and 28.46), with bio-fertilizer 

(Azospirillum) +FYM (15t/ha). INM as whole stimulated the 

assimilation of carbohydrates and proteins which in turn 

enhanced the cell division and cell elongation in the regions 

of auxiliary buds. Similar results have also been reported by 

Rafi et. al., 2002 and Prabhu, et. al., 2003. Increased number of 

branch/plant by application of inorganic fertilizer and biofertilizer 

was also observed by Dange, et. al., 2003. In respect 

of number of branches and number of leaves per plant, the 

maximum number of branches and leaves/plant reported in 

the plant treated with 50% RDF +Azotobacter + 1300ppm 

Cycocel with export quality fruits in okra Nawalkar, et. al. 

2007. 

Number of fruits per plant : 

Biofertilizers and FYM with reduced dose of RDF have 

significant influence on marketable green fruit yield of okra 

(Table 1). The highest number of fruits (21.13/plant) were 

obtained from treatment with biofertilizer (Azotobacter+ 

Azospirillum + PSB) +FYM (15t/ha) +RDF (50%) followed by 

(16.44/plant) in biofertilizer (Azotobacter) +FYM (15t/ha) +RDF 

(50%). However, the minimum number of fruits (11.65/plant) 

were obtained with the treatment biofertilizer (PSB) + FYM 

(15t/ha). Similar findings of significantly higher number of 

fruits/plant by integrated application of chemical fertilizer, 

organic manures and bio-fertilizers have also been reported 

by Prabhu, et. al., 2003 in okra. 

Fruit length (cm) 

Significant effects on variation in fruit length have been 

recorded due to different treatment. The longest fruit (11.62cm) 

were obtained with biofertilizer (Azotobacter+ Azospirillum 

+ PSB) +FYM (15t/ha) +RDF (50%) followed by (10.79cm) in 

Table 1. 

Treatments 

Effect of nutrient management on growth & yield attributing characters of okra 

Plant height 

(30 DAS) 

cm 

Plant height 

(60 DAS) cm 

Number of 

branch /plant 

Number of 

leaves/plant 

Number of 

fruit/plant 

Fruit 

length (cm) 

Fruit 

diameter 

(cm) 

Fruit 

yield/plot 

(kg) 

Fruit yield 

(q/ha) 

T 1 44.46 57.54 3.47 33.03 15.85 10.53 1.70 6.60 132.00 

T 2 45.12 56.80 3.15 31.97 13.99 8.87 1.49 7.06 141.20 

T 3 42.14 54.88 2.89 28.46 12.63 8.32 1.42 6.26 125.27 

T 4 41.85 51.59 2.58 26.15 11.65 7.27 1.35 5.56 111.20 

T 5 46.63 70.91 3.21 34.73 16.44 10.79 1.76 8.07 161.47 

T 6 43.02 56.08 3.12 31.88 15.07 9.58 1.57 7.41 148.27 

T 7 43.65 54.96 2.99 31.45 14.38 8.34 1.45 7.26 145.27 

T 8 46.98 72.05 3.97 41.32 21.13 11.62 1.88 9.76 196.07 

T 9 44.14 55.24 3.31 32.82 15.19 9.35 1.49 8.14 162.80 

SEm (±) 0.560 0.944 0.088 0.559 0.417 0.318 0.029 0.205 0.204 

CD (P=0.05) 1.188 2.001 0.187 1.184 0.886 0.674 0.063 0.434 0.433

PRASAD & NAIK : Effect of Varying NPK Levels and Bio-fertilizers on Growth and Yield of Okra 169 

treatment with biofertilizer (Azotobacter) +FYM (15t/ha) +RDF 

(50%). However, the shortest fruit (7.27cm) were obtained by 

the treatment biofertilizer (PSB) + FYM (15t/ha). Similar 

significant effects with longest fruits have also been reported 

by Kabura, et al. (2001) with the application of organic manures 

and bio-fertilizers. 

Fruit diameter (cm) 

Significantly maximum fruit diameter (1.88cm) were 

obtained by biofertilizer (Azotobacter+ Azospirillum + PSB) 

+FYM (15t/ha) +RDF (50%) followed by (1.76) with biofertilizer 

(Azotobacter) +FYM (15t/ha) +RDF (50%). However, 

significantly the minimum diameter of fruit were obtained by 

the treatment with bio-fertilizer (PSB) + FYM (15t/ha) (1.35cm). 

Better fruit diameter of okra was due to integrated application 

of 50% RDF + organic manure +Biofertilizer have also been 

reported by Naidu, et al., 2002. 

Fruit yield /plot (kg) 

The crop yields/plot varied significantly due to different 

treatments. The yields were increased by 5.56 kg to 9.76 kg 

per plot due to application of graded recommended dose of 

fertilizer (50%) levels with FYM (15t/ha), biofertilizer or both. 

Application of biofertilizer (Azotobacter+ Azospirillum + PSB) 

+FYM (15t/ha) +RDF (50%) significantly produced the highest 

fruit yield per plot (6.76 kg/plot) followed by (8.07 kg/plot) 

with application of biofertilizer (Azotobacter) +FYM (15t/ha) 

+RDF (50%). However, lowest yield of fruits/plot were obtained 

by the treatment of biofertilizer (PSB) + FYM (15t/ha) (5.56kg). 

Similar results have also been found by Gowda and Gowda, 

1983 in okra. 

Fruit yield (q/ha) 

The crop yields due to application of graded 

recommended dose of chemical fertilizer (50%) levels with FYM 

(15t/ha), biofertilizers or both were varies significantly and 

increased by 111.20 to 196.07 (q/ha). Application of biofertilizer 

(Azotobacter+ Azospirillum + PSB) +FYM (15t/ha) 

+RDF (50%) significantly produced maximum fruit yield 

(196.07q/ha) followed by (161.47q/ha) with bio-fertilizer 

(Azotobacter) +FYM (15t/ha) +RDF (50%) yield. However, 

significantly minimum fruit yield (111.20q /ha) were obtained 

by the application of biofertilizer (PSB) + FYM (15t/ha). Similar 

reports of significantly higher total fruit yield due to integrated 

application of organic and inorganic sources of nutrients than 

the sole application of inorganic fertilizers have also been 

reported in okra by Patil, et al., 2000 and Ray, et al., 2005. 


Anburani, A. and Manivannan, K. 2002. Effect of intergrated nutrient 

management on growth in brinjal (Solanum melongena L).cv. 

annamalai. S. Indian Hort., 50(4-6): 377-386. 

Arora, S.K.; Kumar, N. and Sharma, B.R. 1991. Effect of nitrogen and 

phosphorus fertilizers on growth and yield component in okra 

(Abelmoschus esculentus (L.)Moench). Haryana J. Hort., 20 

(3-4): 261-266. 

Barani, P. and Anburani, A. 2003. Influence of vermi composting on 

major nutrients in bhindi (Abelmoschus esculentus (L.) 

Moench).var.Arka Anamika. S. Indian Hort., 52(1-6): 351-354. 

Dange, R. G.; Naik, D. M. and Prabhu, T. 2002. Effect of organic and 

inorganic fertilizers on growth, yield and quality of chilli ( Capsicum 

annum L.) S. Indian Hort., 50(4-6): 578-583. 

Gowda,N. N.C.and Gowda, P.M. 1983. Effect of inter row and cycocel 

on growth and yield of Bhindi. S. Indian Hort., 31 (4/5): 210-214. 

Kabura, B.H.; Kwari, J.D. and Mainu, I. 2001. Response of okra varieties 

to FYM in the semi- arid zone of northeastern Nigeria. J. Sustain. 

Agric. Envrt. 3(1): 63-69. 

Naidu, A.K.; Kushwah, S.S. and Dwivedi, Y.C. 2002. Influence of organic 

manures, chemical and biofertilizers on growth, yield and economics 

of brinjal. S. Indian Hort., 50(4-6): 370-376. 

Nawalkar, L.R.;Khiratkar, S.D.;Badge, S.A.; Chopde,N.K. and Dadgal, 

S.S. 2007. Effect of biofertilizers and growth regulator with reduced 

doses of NPK on growth and yield of okra. J. Soils and Crops. 

17(1): 145-149. 

Patil, M.B.; Jogdand, S.D. and Jadhar, A.S. 2000. Effect of organic and 

biofertilizers on yield and quality of okra. J. Maharashtra Agric. 

Univ., 25(2): 213-214. 

Prabhu, T.; Narwadekar, P. R.; Sannindranath, A.K. and Rafi, Mahd. 

2003. Effect of intergrated nutrient management on growth and 

yield of okra. (Abelmoschus esculentus (L.)Moench) cv. Parbhani 

Kranti. The Orissa J. Hort., 31(1): 17-21. 

Rafi, Mahd.; Narwadekar, P.R.; Prabhu, T. and Sannindranath, A.K. 

2002. Effect of organic and inorganic fertilizers on yield and quality 

of Tomato( Lycopersocon esculetum Mill.) J. Soils and Crops., 

12(2): 167-169. 

Roy, R., Patra, S.K., Ghosh, K.K. and Sahoo, S.K. 2005. Intergrated 

nutrient management in okra(Abelmoschus esculentus (L.)Moench) 

in a river basin. Indian J. Hort., 62(3): 260-264. 



Breeding Periodicity of the Mullet, Liza macrolepis from Mangalore Waters 

H.N.ANJANAYAPPA 1 , S. BENAKAPPA, S.M. SHIVAPRAKASH, S.R. SOMASHEKARA, A. S. KUMAR 

NAIK, JITENDRA KUMAR, MAHESH, V. 

Dept. of Fisheries Resources and Management, Karnataka Veterinary, Animal and Fisheries Sciences University, 

College of Fisheries, Mangalore 575 002 

e-mail: anjanayappahn@rediffmail.com, jitenderanduat@gmail.com 

ABSTRACT 

Studies on breeding periodicity of Liza macrolepis inhabiting 

Mangalore waters indicated that this species spawns only once 

in a year over a period extending from June to November with 

a peak during July to August. Data on the size at first maturity 

of Liza macrolepis using cumulative frequency method showed 

that male attained first maturity at 223.64 mm T.L., while the 

female at 236.36 mm T.L. Male always recorded lower Gonado- 

Somatic Index (GSI) values than female due to higher ovary 

weight compared to testis. The GSI values were high during 

June to November with peaks in July and August indicating 

the spawning period of this fish. The fecundity of fish ranged 

from 1, 22,278 to 6, 74,232 eggs, with an average of 2, 84,146 

eggs. Log linear relationships were established between 

fecundity and total length, fecundity and body weight of the 

fish as well as fecundity and gonad weight. The sex-ratio of 

male to female was 1:0.85. There was an overall male dominance 

in the population. 

Key words Breeding periodicity, Liza macrolepis, Mangalore. 

Mullets form one of the important fisheries of the 

estuaries and coastal waters of India.Though the catch of 

grey mullets is negligible (0.24%) in the total marine fish 

production of India, they are highly valuable and are in great 

demand. There is also a considerable scope to increase their 

production through aquaculture practices like extensive and 

intensive culture, poly or monoculture. The mullet, Liza 

macrolepis is one of the most important species along 

Dakshina Kannada coast, very little information is available 

on the spawning area, time of spawning and other related 

parameters hence, the present study was undertaken to 

investigate spawning, fecundity and sex-ratio of this species 

from Mangalore region. It is hoped that this information would 

help in rational exploitation and management of Liza 

macrolepis along this part of the coast. 


The study is based on the random samples totaling to 

1100 individuals in the size range of 95 to 300 mm total length 

(TL) comprising 594 male and 506 female. Fortnightly samples 

were collected from the markets and commercial fish landing 

centres along the Mangalore coast during the period between 

April 2003 and March 2004. The commercial catches are landed 

by using cast nets, seine nets and gill nets from the estuaries 

of Netravati, Gurpur, Pavanje and Shambhavi. 

In the laboratory, the total length (mm), weight (g), sex 

and maturity stage of individual fish were noted. The ovaries 

of matured females were preserved in 5% formalin for further 

studies. Six stages of maturity (immature, maturing, early 

mature, late mature, gravid and spent) were recognized on the 

macroscopic appearance of the ovary and microscopic 

characteristics of ova. Eggs were measured by an occular 

micrometer. Frequency polygons were drawn for all the stages 

of maturity to find out the frequency of spawning. Gonado 

Somatic Index (GSI) was calculated by using the formula, 

gonad weight x 100/ fish weight. Size at first maturity (50%) 

was determined by using fish with gonads from stage IV 

onwards and the relative condition factor (K n 

) values at various 

size groups. Fecundity was estimated by using ovaries of 

stages IV and V. Sex-ratio was calculated for different months 

and different size groups of fish. 


Development of ova to maturity: 

Ova diameter frequency polygons of Liza macrolepis 

in various stages of maturity are given in Fig. 1.The size of 

ova ranged between 0.02 and 0.64 mm (Table 1).In stage I, the 

size of ova ranged from 0.02 mm to 0.10 mm, majority of them 

ranging in size from 0.02 mm to 0.06 mm. In stage II, a batch of 

maturing eggs is withdrawn from the general egg stock. The 

maturing group had a modal value at 0.06 mm, while the largest 

ova measures at 0.24 mm. In stage III, this progresses to 0.18 

mm, the maximum size being 0.32 mm. The mode at 0.18 mm of 

stage III moves to 0.32 in stage IV with a maximum size at 0.44 

mm. In stage V, there was only one group of mature eggs 

forming the mode at 0.52 mm. However, maximum size of ova 

measured up to 0.64 mm. In stage VI, there was a mode at 0.02 

mm, maximum size of the ova being 0.16 mm. This stage 

resembles I or II stage. The fish seems to remain in this 

condition until the maturation cycle commences again. 

Table 1. 

Stages of 

maturity 

Classification of maturity stages of Liza 

macrolepis 

Description of 

ova 

Mode of largest 

group of ova (mm) 

I Immature 0.02 0.10 

II Maturing 0.06 0.24 

III Early mature 0.18 0.32 

IV Late mature 0.32 0.44 

V Gravid 0.52 0.64 

VI Spent 0.02 0.16 

Maximum size 

of ova (mm)

ANJANAYAPPA et. al.,: Breeding Periodicity of the Mullet, Liza macrolepis from Mangalore Waters 171 

Frequency of spawning: 

It was observed from the ova diameter studies that there 

are two groups of ova, viz., immature and mature in a mature 

ovary. A batch of the immature stock of ova develops to form 

a distinct group of mature eggs to be spawned in the ensuing 

spawning season. The unspawned larger eggs in spent ovaries 

are resorbed as evidenced by the presence of thin shell like 

structure left over in such ovaries. This fish seems to remain 

in this condition until the maturation cycle commences again. 

Hence, it appears that the individual spawns only once a year. 

Single spawning was also reported by Sarojini, 1957 in Mugil 

Fig. 1. 

Ova diameter frequency polygons of Liza macrolepis 

in various stages of maturity. 

Table 2. 

Month-wise percentage of gonads in different 

stages of maturity of Liza macrolepis 

Months No. of fish Sex 

Maturity stages 

I II III IV V VI 

52 M 51.92 32.69 15.38 - - - 

Apr 2003 51 F 47.06 19.61 33.33 - - - 

47 M 40.43 25.53 27.66 6.38 - - 

May 45 F 44.44 28.88 26.67 - - - 

50 M 46.00 26.00 14.00 10.00 4.00 - 

Jun. 49 F 20.41 24.49 26.53 22.45 6.12 - 

41 M 31.71 21.95 19.51 21.95 4.89 - 

Jul. 52 F 7.69 15.38 15.38 28.85 32.69 - 

Aug. 

44 M 20.45 4.55 19.91 29.55 27.27 2.27 

51 F 9.80 11.76 15.69 39.22 21.57 1.96 

Sep. 

50 M 10.00 22.00 14.00 26.00 24.00 4.00 

41 F 21.95 7.32 17.07 34.15 14.63 4.88 

Oct. 

39 M 17.95 33.33 25.64 12.85 7.69 2.56 

26 F 30.77 30.77 19.23 11.54 7.69 - 

Nov. 

54 M 27.78 16.67 18.52 20.37 14.81 1.85 

34 F 35.29 47.06 8.82 5.88 2.94 - 

Dec. 

48 M 54.17 27.08 16.67 2.08 - - 

36 F 61.11 38.89 - - - - 

Jan. 2004 

56 M 44.64 42.86 10.71 1.79 - - 

38 F 52.63 47.37 - - - - 

Feb. 

60 M 68.33 31.67 - - - - 

42 F 61.90 38.09 - - - - 

Mar. 

53 M 45.28 41.51 13.21 - - - 

41 F 53.65 34.15 12.19 - - - 

Cumulative percentage of mature fish 

100 

90 

80 

70 

60 

50 

40 

30 

20 

10 

Male 

Female 

Fig.2. 

0 

90 110 130 150 170 190 210 230 250 270 290 

Total length (mm) 

Size at first maturity of Liza macrolepis 

Fig.3. 

Monthly variation in the Gonado-Somatic Index in Liza 

macrolepis


Table 3. 

parsia and Liza macrolepis and Mugil cephalus by Luther, 

1963 concluded that mullets exhibited non intermittent and 

single spawning per season because of the production of 

only one set of ova per year. 

Spawning season: 

Number of mature ova in individuals of L Liza 

macrolepis 

Sl. Total length Body weight Gonad weight 

Stage of 

Fecundity 

No. (mm) (g) 

(g) 

maturity 

1 215 79 4.18 1,22,278 IV 

2 220 81.5 4.24 1,37,262 IV 

3 225 84.5 5.16 1,51,549 IV 

4 245 129 5.98 2,27,240 IV 

5 250 139 6.12 2,20,320 V 

6 255 125 4.79 1,57,942 IV 

7 265 180 6.81 2,35,444 V 

8 275 187 9.87 2,53,066 IV 

9 275 189 10.45 2,36,170 IV 

10 280 200 12.38 2,58,907 V 

11 285 20 15.96 2,92,919 V 

12 290 207 16.48 3,12,460 V 

13 295 25 10.50 3,46,920 IV 

14 300 300 16.41 6,74,232 V 

15 300 295 14.54 6,35,494 IV 

Percentage occurrence of different stages of gonads of 

Liza macrolepis in different months are given in Table 2. 

Spawning stages of gonads first occurred during the month 

of June. From the Table 2 it is clear that the species Liza 

macrolepis spawned only once in a year over a prolonged 

period extending from June to November with a peak during 

July to August along the Mangalore coast. The gonadosomatic 

index values also revealed (Fig.3) that the spawning 

season extended from June to November. The average GSI 

observed was more in female than in male perhaps due to a 

greater ovary weight compared to testes and subsequent drop 

in GSI indicates the release of gamets. According to Luther, 

1963 the spawning season of L. macrolepis and Mugil cephalus 

in Mandapam waters extended from June to February with a 

peak during July and August. Such extended spawning season 

was also reported in L. tade (Reddy, 1985), V. speigleri 

(Baburaj, 1987) and V. seheli (Moorthy, 2000) from Mangalore 

waters. 

Size at first maturity: 

Data on the size at first maturity of L. macrolepis using 

cumulative frequency method (Fig.2) showed that male 

attained first maturity at 223.64 mm TL. While for female, the 

size at first maturity was 236.36 mm TL. According to the 

observations of Luther, 1963, male and female of L. macrolepis 

attained first maturity at about 160 and 170 mm respectively. 

Jhingran and Natarajan, 1970 reported that the size at first 

maturity for the male and female of M. macrolepis was 212 mm 

and 237 mm respectively. Wijayarathne and Costa, 1987 

reported that L. macrolepis Negombo lagoon, attained 

maturity at 16 and 21 cm TL of in male and female respectively. 

Krishna kumar and Balakrishnan Nair, 1995 worked on breeding 

biology of V. cunnesius inhabiting the Ashtamudi estuary. 

According to them the size at first maturity for male was around 

139 mm and for female 148 mm TL. 

Fecundity 

Knowledge of total number of eggs produced by a fish 

in a year is important for determining the spawning potential 

of a fish. Fecundity is usually determined from the number of 

ova of the mature group in the ovary. The details are presented 

in Table 3. In the present study, fecundity of L. macrolepis 

found to vary from 1,22,278 to 6,74,232 eggs depending upon 

the size of fish (215 mm to 300 mm TL.). Luther, 1963 reported 

that in case of L. macrolepis the fecundity ranged from 1,51,920 

to 6,76,200 eggs. Das 1977b observed the fecundity of grey 

mullet, M. cephalus at 43,000 to 5,13,000 in fishes varying in 

length from 140 to 315 mm. Baburaj, 1987 reported that 

fecundity in V. speigleri varied from 1,34,327 to 8,39,492 for 

the fish ranging between 150 and 415 mm in total length. 

Logarithmic relations of fecundity (F), with length of fish (L), 

weight of fish (W), and gonad weight (GW) are given in 

Table 4. The Logarithmic relations between fecundity and 

length of fish, fecundity and weight of fish and fecundity and 

gonad weight were found to be linear indicating that the 

fecundity generally increased with increasing length, weight 

and gonad. 

Sex ratio: 

Sex-ratio studies (Table 4) indicated that there was an 

overall predominance of male in the population. The sex-ratio 

between male and female was 1.00: 0.85 during the period. 

Chi-square test (5%) showed no significant deviation from 

the theoretical ratio of 1: 1 except November. 

Table 4. 

Logarithmic relationship of fecundity with length 

(L), weight (W), and gonad weight (GW) of Liza 

macrolepis 

Statistical relationship 

Correlation 

Logarithmic form Power equation form coefficient (r) 

F = -3.8713 + 3.8271 L F = -3.8713 + L 3.8271 0.8634 

F = 3.0492 + 1.0604 W F = 3.0492 + W 1.0604 0.9173 

F = 4.6016 + 0.8512 GW F = 4.6016 + GW 0.8512 0.8539 

Study on spawning periodicity of Liza macrolepis 

indicated that this species spawned only once in a year over 

a prolonged period extending from June to November with a 

peak during July to August. Data on the size at first maturity 

of L. macrolepis using cumulative frequency method showed 

that male attained first maturity at 223.64 mm T.L., while for 

female, the size at first maturity was 236.36 mm T.L. Male always 

recorded lower gonado-somatic index (GSI) values than female 

due to greater ovary weight compared to testes. The GSI 

values were high during June to November with peaks in July 

and August indicating the spawning period of this fish along 

Mangalore coast. The fecundity of fish ranged from 1,22,278 

to 6,74,232 eggs, with an average of 2,84,146 eggs. Log linear

ANJANAYAPPA et. al.,: Breeding Periodicity of the Mullet, Liza macrolepis from Mangalore Waters 173 

Table 5. 

Sex ratio in different size groups of Liza 

macrolepis 

Months 

Total no. Male Female Chi – square Male : 

of fishes n N values Female 

Apr. 2003 103 52 51 0.0098 1 : 0.98 

May 92 47 45 0.0434 1 : 0.96 

Jun. 99 50 49 0.0102 1 : 0.98 

Jul. 93 41 52 1.3010 1 : 1.27 

Aug. 95 44 51 0.5158 1 : 1.16 

Sep. 91 50 41 0.8902 1 : 0.82 

Oct. 65 39 26 2.6000 1 : 0.67 

Nov. 88 54 34 4.5454* 1 : 0.63 

Dec. 84 48 36 1.7142 1 : 0.75 

Jan. 2004 94 56 38 3.4468 1 : 0.68 

Feb. 102 60 42 3.1764 1 : 0.70 

Mar. 94 53 41 1.5318 1 : 0.77 

Total 1100 594 506 7.0400 1 : 0.85 

relationships were established between fecundity and total 

length, fecundity and body weight of the fish as well as 

fecundity and gonad weight. The sex ratio of male: female was 

1: 0.85. There was an overall predominance of male in the 

population. 


Baburaj, D. 1987. Some aspects of biology of the mullet, Valamugil 

speigleri (Bleeker) from Mangalore region. M.F.Sc. thesis, Univ. 

Agril. Sci., Bangalore. pp.155. 

Das, H. P. 1977 (b). Length - weight relationship and relative condition 

of grey mullet, M. cephalus. Mahasagar., 10(3-4): 145-149. 

Jhingran, V.G. and Natarajan, A. V. 1970. Study on the fisheries and fish 

populations of the Chilka lake for breeding. Curr. Sci., 27(5): 

181-182. 

Krishnakumar, K. and Balakrishnan Nair, N.B. 1995. Breeding biology 

of the grey mullet Valamugil cunnesius from the Astamudi estuary, 

South-west coast of India. Fish. Technol. Soc. Fish. Technol (India)., 

32(1): 1-5. 

Luther, G. 1963. Some observations on the biology of Liza macrolepis 

(Smith) and M. cephalus Linnaeus (Mugilidae) with notes on the 

fishery of grey mullets near Mandapam. Indian J. Fish., 10: 642-665. 

Moorthy K.S.V. 2000. Feeding ecology and population characteristics 

of the mullet, Valamugil seheli (Forskal) from Mangalore coast. 

Ph. D. thesis, Univ. Agril. Sci., Bangalore. pp.190 

Reddy, S. 1985. Some aspects of biology of the mullet, Liza tade (Forskal) 

from Mangalore region. M.F.Sc. thesis, Univ. Agril. Sci., Bangalore. 

pp.125. 

Sarojini, K.K. 1957. Biology and fisheries of the grey mullets of Bengal. 

1. Biology of Mugil parsia (Hamilton) with notes on its fishery in 

Bengal. Indian J. Fish., 4: 160-205. 

Wijeyaratne, M.J.S. and Costa, H.H. 1987 (b). The food, feeding and 

reproduction of the Borneo mullet, Liza macrolepis (Smith) in a 

coastal estuary in Srilanka. Indian J. Fish., 34(3): 283-291. 



Effect of Biofertilzers on Growth and Yield Attributes of Pea (Pisum sativum L.) 

PUSHKAR SINGH PATEL, R.B. RAM, JAYPRAKASH AND M.L.MEENA 

Department of Applied Plant Science (Horticulture),Babasaheb Bhimrao Ambedkar (Central University), 

Vidya Vihar Raebareli Road, Lucknow, Uttar Pradesh 226 025 

e-mail: singhjayprakash94@gmail.com 

ABSTRACT 

A field experiment was carried out to assess the growth and 

yield characters of pea cv Arkel. The experiment was laid out in 

randomized block design with three replications for pea crop 

consisting 15 treatments namely, RDF of NPK (25: 70: 50 ) kg/ 

ha, Azospirillum+50% N and RDF of P and K, Azospirillum+ 

75% N and RDF of P and K, Azospirillum+ RDF of NPK, PSB+ 

50% P and RDF of N &K, PSB+ 75% P and RDF of N&K, PSB+ 

RDF of NPK, Rhizobium+50% N RDF of N&K, Rhizobium+ 

75% N and RDF of P&K, Rhizobium+ RDF of NPK, VAM+ 50% 

P and RDF of N&K, VAM+ 75% P and RDF and N&K, VAM+ 

RDF of NPK, PSB+ Rhizobium +50% NPK, PSB+ Rhizobium+ 

50% NP & full dose of K to find out the effect of bio fertilizers, 

inorganic and its combinations on days to germination, number 

of plants per plot, plant height (cm), days to flowering, number 

of flowers per plant, days to first fruit harvest, number of pods 

per plant, number of seeds per pod, length of leaf (cm), shelling 

percentage, weight of 10- pods (g), seed weight (g), yield per 

plot, length of pod (cm).However, The application of inorganic 

and biofertilizers, treatment – T 1 

- (Rhizobium and RDF of 

NPK), showed superior among all treatments under 

investigation. 

Key words Biofertilizer, Pea, VAM, Rhizobium, Azospirillum 

The progressive use of fertilizers along with inorganic 

fertilizer may be the right answer in the recent year and use of 

microbial inoculants as source of biofertilizers have become a 

hope for most of the countries as for as economically and 

environmental point of voice are concerned. Bio-fertilizers are 

economically, attractive and ecologically sound means of 

fertilization. Therefore, is developing countries like India it 

can solve the problem at highest cost of fertilizers and help in 

saving the economy of the country. The biofertilizers denote 

all the nutrients are part of biological origin for plant growth. 

They passes unique ability to enhance productivity by 

biological nitrogen fixation are solubilization at insoluble 

phosphate or producing hormones vitamins are other growth 

factor require for plant growth. The beneficial microbes which 

are of great significant are biological nitrogen fixation 

phosphate solubilizers and mycorrhizal fungi Azotobacter is 

the bacterial which have been known to fix nitrogen 

biologically use of these microbial fertilizer cut down the 

quantity of nitrogenous fertilizer with some improvement in 

the crop field. They are also reported to have an effective role 

in improving disease resistant in the crop by producing anti 

bacterial and fungal compounds and also produce growth 

regulators. Azospirillum is a group of bacteria which fixes N 2 

in loose association with plant. It fixes nitrogen equivalent to 

40-50 Kg/ ha and also produces IBA, GA 3 

and cytokinins like 

substances. It also increases root biomass in the inoculated 

plants, thereby it helps in greater absorption of nutrients. As 

well as resulting is higher yield. Around 95 to 99% of the total 

soil phosphorus is insoluble which is not directly available to 

plants. The phosphate solubilizing bacteria may convert it 

into soluble form for by producing organic acids about 15 to 

25% of insoluble phosphate can be solubilized saving chemical 

fertilizers significantly. The endomycorrhizae otherwise known 

as the vesicular arbuscular mycorrhizae (VAM) are the most 

ubiquitous fungal association among angiosperms. The 

importance of VAM fungi is well appreciated due to their ability 

to improve nutrient uptake, disease resistant and drought 

resistant etc. 


The present investigation entitled “Effect of bio 

fertilizers on growth and yield traits of pea (Pissum sativum L 

sp. hortense) cv. Arkel” was carried out at the Horticulture 

Research Farm of the Department of Applied Plant Science 

(Horticulture), Babasaheb Bhimrao Ambedkar University, 

Vidya Vihar, Rae Bareli Road Lucknow (U. P.) in randomized 

block design with three replications in plot size of 1.5 x1.0 m 

with spacing of 30 x10 cm during Rabi season of 2011-12. 

Experimental field was laid out in randomized block design 

with 15 treatments and replicated thrice. The treatment 

combinations comprised namely, T 1 

- RDF of NPK (25:70:50) 

kg /ha, T 2 

- Azospirillum + 50% N and RDF of P&K, T 3 

- 

Azospirillum +75% N and RDF of P&K, T 4 

- Azospirillum + 

RDF, T 5 

- PSB+ 50% P and RDF of N&K, T 6 

- PSB+ 75% P and 

RDF of N&K, T 7 

- PSB+ RDF, T 8 

- Rhizobium+ 50% N and RDF 

of P&K, T 9 

- Rhizobium+ 75% N and RDF of P&K, T 10 

- 

Rhizobium+RDF, T 11 

- VAM+50% P and RDF of N&K, T 12 

- 

VAM+ 75% P and RDF of N&K, T 13 

- VAM+ RDF, T 14 

- 

PSB+Rhizobium + 50% NPK, T 15 

- PSB+ Rhizobium+ 50% NP 

and full dose of K. Other cultural practices like mulching, 

irrigation, hoeing, insect-pest and disease management were 

common for the each treatment. Observations were record on 

14 characters including vegetative and yield attributing 

characters viz., days to germination, number of plants per 

plot, plant height (cm), days to flowering, number of flowers

PATEL et. al., : Effect of Biofertilzers on Growth and Yield Attributes of Pea (Pisum sativum L.) 175 

per plant, days to first fruit harvest, number of pods per plant, 

number of seeds per pod, length of leaf (cm), shelling 

percentage, weight of 10- pods (g), seed weight (g), yield per 

plot, length of pod (cm).The data were subjected to statistical 

analysis to test the level of significance. 

RESULTS AND DISCUSION 

All parameters viz., showed a significant increase 

(Table 1 and 2) which the application of biofertilzers in various 

combinations. Number of plants per plot-The observations 

recorded in case of number of plants per plot in various 

treatment found significant but there are maximum number of 

plants under treatment T 15 

- (PSB+ Rhizobium + 50% NPK), 

and minimum number of plant per plot under treatment T 1 

- 

(RDF of NPK). The probable reasons for such results could 

be because of certain growth promoting substances secreted 

by the microbial inoculants, which is turn might have led to 

better root development, better transpiration of water, uptake 

and deposition of nutrients similar observations have also 

been made by other workers (Kandasamy, et al., 1985) in 

various crops. Days of germination- The observation in case 

of days to require to germination among various treatments 

were found significant. The maximum days to germination 

were in treatment T 5 

- (PSB+ 50% P and RDF of N&K), and 

minimum day to require for germination under treatment T 11 

- 

(VAM+ 50% P and RDF of N and K). Number of flowers per 

plant. The maximum flowers are found under treatment T 7 

- 

(PSB+ RDF of NPK), and minimum flower observed under 

treatment T 1 

- (RDF of NPK), (Bisen, et. al., 1985) also found 

maximum number of flower on the application of PSB in brinjal 

and chilli. Days to flowering- The maximum days required to 

flowering observed under treatment T 1 

- (RDF of NPK), and 

minimum days to required to flowering T 7 

- (PSB+ RDF of NPK). 

Plant height (cm) - the maximum plant height was shown 

constantly by the treatment T 10 

- (Rhizobium+RDF of NPK), 

The leaf grown was studied and it was found that leaf length 

showed significant result due to different bio fertilizers 

treatments. The treatment T 11 

- (VAM+ 50% P and RDF of N&K), 

showed the maximum leaf length and development of vigorous 

plant width and larger leaf size may be due to the synthesis of 

grown promoting substances by the PSB. The similar result 

were reported by (Chattoo, et. al., 1997), in case of knoll kohl. 

Although Azotobactor showed a better performance but was 

not comparable with Azospirillum in aspect of growth 

attributes. The probable reasons for such result could be 

because of certain growth promoting substances secreted by 

the microbial inoculants, which is turn might have led to better 

root development, better transpiration of water, uptake and 

deposition of nutrients similar observations have also been 

made by other workers (Kandasamy, et. al., 1985), in various 

crops treatment T 1 

- (Recommended dose of NPK though 

chemical fertilizers) showed the minimum number of leaves 

per plant at45 DAT but from 75 DAT onwards , the minimum 

number of leaves per plant was observed under treatment T 1 

- 

( Recommended dose of NPK though chemical fertilizers ), up 

to 90 DAT. Length of pods –the maximum length of pods was 

observed under treatment T 7 

- (PSB+ RDF of NPK), and 

minimum length of pods observed under treatment T 1 

- (RD of 

NPK). Number of pods per plant–the maximum number of 

pods per plant was recorded in case of treatment T 13 

- (VAM+ 

RDF of NPK), and minimum number of pods per plant under 

treatment T 1 

- (RDF of NPK). Number of pods was directly 

related to the number of flowering of plants. No. of seed per 

pods-the observation recorded in case of number of seeds 

per pod was maximum under treatment T10- (Rhizobium+ RDF 

of NPK), and minimum seed per pod under treatment T 1 

- (RDF 

of NPK). Days to first fruit harvest- the maximum days of first 

pod harvest observed under treatment T 10 

- (Rhizobium+ RDF 

Table 1. Effect of bio fertilizers on vegetative characters of pea. 

S.N. Treatments Parameters 

No. of plant 

per plot 

Days to germination 

(50%) 

No. of flower 

per plant (50%) 

Days to 

flowering 

Plant height 

(cm) 

Length of leaf 

(cm) 

Length of 

pods (cm) 

1 T 1 41.120 7.670 8.220 36.650. 45.890 5.860 7.150 

2 T 2 43.670 8.920 10.110 41.120 47.890 7.980 8.120 

3 T 3 44.890 7.170 12.240 38.780 49.780 8.120 8.450 

4 T 4 45.780 8.020 14.780 41.170 55.830 9.080 8.720 

5 T 5 44.550 6.190 11.120 41.980 46.320 8.170 7.980 

6 T 6 46.180 7.180 13.560 43.670 47.820 9.810 8.280 

7 T7 46.980 6.780 15.170 44.920 52.320 9.970 8.880 

8 T 8 45.180 8.180 11.780 39.190 54.720 7.790 7.860 

9 T9 46.280 6.380 13.680 42.370 53.120 8.130 8.170 

10 T 10 47.070 7.790 14.520 44.890 56.390 9.170 8.290 

11 T 11 45.130 9.120 10.870 40.350 50.240 6.890 7.950 

12 T 12 46.380 7.890 12.890 42.270 52.360 8.160 8.080 

13 T 13 46.830 7.110 13.820 39.650 54.630 9.340 8.380 

14 T 14 46.860 8.840 12.870 41.170 51.910 8.120 7.980 

15 T 15 47.780 6.953 14.650 42.340 55.810 8.980 8.040 

C.D.(P= 0.05) 1.538 0.087 0.227 0.184 1.337 0.189 0.061 

S.E.m± 0.744 0.068 0.092 0.100 0.648 0.104 0.045


Table 2. 

Effect of bio fertilizers on yield and yield attributing traits of pea. 

S.N. 

Treatments 

Parameters 

No. of pods per 

plant 

No. of seeds 

per pod 

Days to first 

fruit harvest 

Shelling (%) 

Weight of ten 

pods (g) 

100-Seed 

weight (g) 

1 T 1 8.340 5.890 50.130 58.000 52.890 20.020 428.000 

2 T 2 9.130 5.920 52.120 60.000 53.120 20.340 490.000 

3 T 3 9.980 6.120 52.890 61.120 54.340 20.980 515.000 

4 T 4 10.340 6.340 53.870 61.340 54.450 21.000 590.000 

5 T 5 8.940 5.940 52.130 60.340 53.000 20.000 510.000 

6 T 6 9.120 6.340 53.410 60.980 53.830 21.890 580.000 

7 T 7 10.780 6.810 55.430 61.500 54.120 22.080 621.000 

8 T 8 9.180 6.120 52.810 59.120 53.020 19.980 480.000 

9 T 9 9.870 6.670 54.160 60.340 53.890 20.340 528.000 

10 T 10 11.120 7.120 56.230 61.120 54.050 20.980 630.000 

11 T 11 9.350 5.970 52.350 60.670 53.120 19.970 5000.000 

12 T 12 10.120 6.340 53.830 60.980 53.980 20.870 580.000 

13 T 13 11.920 6.780 55.820 61.860 54.250 21.100 620.000 

14 T 14 10.390 6.030 54.320 60.080 53.830 20.120 510.000 

15 T 15 10.890 6.820 55.120 60.870 54.120 20.920 610.000 

C.D.(P=0.05) 0.092 0.134 1.177 1.489 0.934 0.437 2.596 

S.E.m± 0.033 0.157 0.572 0.726 0.456 0.200 1.267 

Yield per plot(g) 

OF NPK),and minimum days to be required for first pod harvest 

under treatment T 1 

- (RDF of NPK). Shelling percentage- It 

was observed that, minimum shelling percentage under 

treatment T 1 

- (Rhizobium+ RDF of NPK), and maximum 

shelling percentage under treatment T 13 

-(VAM+ RDF of NPK). 

Weight of 10- pods –As per data recorded it was found that, 

the weight of ten pods was maximum in the case of treatment 

T 14 

- (Azospirillum + RDF of NPK), while the same was found 

to be minimum under treatment T 1 

- (RDF of NPK). 100 Seed 

weight in the study on pea should maximum seed weight is 

under treatment of T 7 

- (PSB+ RDF of NPK), and minimum 

loading seed weight is observed under treatment T 11 

- (VAM+ 

% P and RDF of NPK). Yield per plot- the maximum yield per 

plot was recorded under the trial treatment T 15 

- (PSB+ 

Rhizobium + % NP and full dose of K), while the minimum 

yield was shown to be under treatment T 1 

-(RDF of NPK). 

LITECTURE CITED 

Bisen , R.K.; Chaubey, P.C.; Pandey, B.R. and Asati, K.P. 1985. Influences 

of nitrogen and spacing of growth and green pod of pea. JNKVV 

Res. J., 19: 68-70. 

Chattoo. M.A.; Gandoo, M.Y. and Zargar, M.Y. 1997. Effect of 

azospirillum and azotobactor on growth yield and quality at knolkhol 

(Brassica oleracia var. gongigloses L.). Veg. sci., 24(1): 16-19. 

Kandasamy, D.G.; Mohanraj Samuel and Oblisamy, G. 1985. Influence 

of VAM and phoshphobacteria on the growth of brinjal and chilli in 

nursery. South Indian Hort., 32: 170-175. 



Studies of Quality Characteristics in Short Grain Scented Rice (Oryza sativa L.) 

Varieties Accessions 

SUNITA KUMARI 1 , R.N.KEWAT, R.P.SINGH AND PRATIBHA SINGH 

Department of Biochemistry, N.D.University of Agriculture & Technology, Kumarganj, Faizabad 

e-mail: sunitafzd@gmail.com 1 

ABSTRACT 

The present investigation on Studies of quality characteristics 

in short grain scented rice varieties/accessions was conducted 

at Crop Research Station, Masodha, Faizabad during kharif 

season. Twelve varieties/strains namely V1, (NDR 6279), V2 

(NDR 6265), V3 (Badshah Bhog), V4 (NDR 6257), V5 (Badshah 

Pasand), V6 (Bhanta Phool A), V7 (IET 19800), V8 Kalanamak A, 

V9 (Kanak Jeer), V10 (Rambhog B), V11 (Kalanamak Berdpur) 

and V12 (Lalmati) were screened out on the basis of physical 

and biochemical traits. Maximum kernel length was observed 

in (7.07 mm ) NDR 6265 and kernel breadth was (1.81 mm) in 

NDR 625. Maximum elongation ratio was observed in (1.88mm) 

Kankjeer and Banta Phool A and kernel length after cooking 

was maximum in (11.4mm) NDR 6265. Maximum carbohydrate 

content was found in Kalanamak Berdpur (79.4%). Maximum 

amylose content was found in variety Kalanamak Berdpur 

(19.8%). Highest total sugar content and non reducing sugar 

content was Badshah Pasonda 1.92 g/100g and 1.72g/100g 

respectively whereas reducing sugar content was highest in 

Kalanamak Berdpur .32 g/100g. On the basis of above 

parameters variety Kalanamak Berdpur, Badshah pasonda, NDR 

6265 and NDR 625 were rated superior among the all varieties/ 

accessions tested in the present investigation. 

Key words Aromatic rice, kernel length, kernel breadth, Elongation 

ratio, Carbohydrate, Amylose and Sugars content. 

Aromatic rice (Oryza sativa L.)contains several 

biochemicals, but the most significant one is identified as 2- 

acetyl pyrroline (2AP). The popcorn-like smell of aromatic 

rice, stemming primarily from its 2-acetyl-1-pyrroline content 

(Bhattacharjee, et al., 2002) or pandan (Pandanus 

amaryllifolius)-like odour. Aromatic rice fetches high prices 

in some international markets including south Asia, the Middle 

East, and particularly India, Pakistan, and Thailand (Kaosaard 

and Juliano, 1992). Thailand, India, and Pakistan are 

competitive producers and developers of aromatic rice in the 

world. However, many of farmers in these countries grow 

specific varieties mainly for export markets. One of the reasons 

is the limitation in aromatic rice production is yield 

improvement. The first high-yielding Basmati rice cultivars 

are Pusa Basmati1 and Kasturi; they yield 4.5 and 4.0 ton/ha 

(about 1.5 and 1.0 tons/ha) higher than traditional Basmati 

varieties (Bhattacharjee, et al., 2002). Until 2007, it was 

accounted for 40- 60% of Basmati rice exports from India 

(Siddiq, et al., 2012). 

There are significant cultural differences in quality 

preferences and the most important acceptance factors for 

Asian consumers living in the United States are cooked rice 

appearance and aroma (Meullenet, et al., 2001). Starch is the 

major constituent of milled rice at about 90% of the dry matter. 

Cooking characteristics, texture, water absorption ability, 

stickiness, volume expansion, hardness and even the 

whiteness and gloss of the cooked milled rice are affected by 

the amylose content (Juliano, 1985b). Carbohydrate is one of 

the major constituents of rice and it also affects their cooking 

qualities which reflect its functional property in rice. Keeping 

in view of above facts, the present investigation was 

conducted. 


The field experiment was conducted during kharif, 

season in completely randomized block design with three 

replications at Crop Research Station Masodha, Faizabad 

(U.P.). All the agronomical practices were done to achieve 

good crop in relation to biochemical content. Healthy scented 

rice seeds were hulling and milling in CRS, Masodha by hulling 

and milling machine. After that all the varieties were brought 

to the laboratory of Biochemistry in N.D.U.A.&T. for 

biochemical analysis. After threshing all varieties were brought 

to laboratory of biochemistry for biochemical analysis. 

Vernaearcalipers was used for calculation of length and 

breadth. Carbohydrate content in grain was determined by 

the method of (Mc cready, et al., 1950). Amylose content in 

grain was determined by the method of Juliano 1979. Total 

sugar content in grain was determined by the method of 

Dubois, et al., 1950. Reducing sugar content in grain was 

determined by the method of Miller, 1959. The non –reducing 

sugar was obtained by subtraction of reducing sugar from 

total sugar. 

Non- reducing sugar =Total sugar –reducing sugar 


Kernel length, Kernel breath, elongation ratio and kernel 

length after cooking have been represented in Table 1. The 

kernel length in varieties ranged from 3.96-7.07 mm. Maximum 

kernel length was found 7.07 mm in NDR 6265 which was 

significantly higher over the rest varieties. Kernel length of 

variety is governed by its genetic potential. Hence, varieties


Table 1. 

Varieties 

Fig.1. 

Table 2. 

Physical characteristics of different varieties/ 

accessions of short grain aromatic rice kernel 

Kernel length 

(mm) 

Kernel 

breadth 

(mm) 

Elongation 

ratio 

Kernel 

length after 

cooking 

Treatment 

NDR 6279 6.30** 1.78** 1.44* 9.12 

NDR 6265 7.07*** 1.66 1.61 11.4*** 

3.96* 1.57 1.68 6.68* 

NDR 625 5.67 1.81*** 1.56 8.86 

5. 64 1.69 1.63 9.12 

Bhanta Phool A 4.26 1.64 1.88*** 8.02 

IET-19800 5.05 1.44 1.65 8.37 

Kalanamak A 5.47 1.72 1.70 9.34 

Kanak-Jeer 5.07 1.60 1.88*** 9.54 

Ram-Bhog-B 5.6 0 1.62 1.59 9.85** 

Kalanamak Berdpur 5.19 1.48 1.72 8.92 

Lalmati 5.36 1.33* 1.75** 9.43 

CD at 5% 0.770 0.220 0.205 1.388 

SEm± 0.264 0.075 0.070 0.475 

Varieties 

Treatment 

Physical characteristics of different varieties/strain of 

short grain aromatic rice kernel 

Variability in carbohydrate, Amylose and sugar 

content of different varieties/strains of short grain 

aromatic rice 

Carbohydrate 

content 

(%) 

Amylose 

content 

(%) 

Total 

sugar 

content 

(g/100g) 

Reducing 

sugar 

content 

(g/100g) 

Nonreducing 

sugar 

(g/100g) 

NDR 6279 67.85 21.78 1.72 0.26 1.46 

NDR 6265 69.53 22.30 1.66* 0.21 1.45 

Badshah-Bhog 78.69** 23.77 1.69 0.24 1.45 

NDR 625 73.69 22.75 1.91 0.22 1.69 

Badshah-Pasonda 69.09 21.62 1.98*** 0.26 1.72*** 

Bhanta-Phool A 64.28* 21.93 1.77 0.18* 1.59 

IET 19800 67.44 20.15 1.72 0.25 1.47 

Kalanamak A 77.61 23.97*** 185 0.20 1.65 

Kanak-Jeer 71.42 21.78 1.75 0.31** 1.44* 

Ram-Bhog B 68.07 21.62 1.86 0.29 1.57 

Kalanamak- 

Berdpur 

79.54*** 22.92 1.94 0.32*** 1.62 

Lalmati 72.27 19.81* 1.97** 0.27 1.70** 

CD at 5% 12.584 3.213 0.329 0.025 0.245 

SEm± 4.311 1.100 0.113 0.008 0.084 

Fig. 2. 

Variability in carbohydrate, Amylose and sugar content 

of different varieties/strains of short grain aromatic rice. 

differed significantly with each other in respect of kernel 

length. The result is supported by Trivedi, 2007 evaluated 

maximum length of rice kernel was found in Pusa Basmati 7.5 

mm. The results are close favors with Singh, et. al., 2005 as 

found the variation in kernel length in range from 4-5.60 mm 

for forty Kalanamak germplasm. 

The kernel breadth in varieties ranged from 1.33-1.81 

mm. maximum kernel breadth was found 1.81 mm in NDR-6265 

which was significantly more over the rest varieties. In respect 

of undertaken varieties differ significantly with each other in 

respect of kernel breadth. The result is supported by Trivedi, 

2007 examined the kernel breadth from 1.74-1.94 mm whereas 

Singh, et al., 2005 witnessed in range from 1.5-2.1 mm. 

Elongation ratio in short grain scented rice ranged from 

1.44-1.88. maximum elongation ratio was found 1.88 in varieties 

Bhonta-Phool A and Kanak-Jeer which was better over the 

rest of the varieties. This result was supported by Singh, 

et al. (2005). 

Kernel length after cooking ranged from 6.68-11.40 mm. 

Maximum kernel length after cooking was found 11.40 mm in 

NDR 6265 which was significantly higher over the rest 

varieties. Similar findings have also been reported by Horuichi, 

1966 and Singh, et al. 2005. 

Data pertaining to carbohydrate content, amylose 

content, total sugar content, reducing sugar content and non 

reducing sugar content have been shown in Table 2.The 

carbohydrate content in various short grain scented rice 

varieties varies from 64.28-79.54 per cent. Maximum 

carbohydrate content was observed 79.54 per cent in variety 

kalanamak- Berdpur. These results have been supported by 

Mangkaeo et al. (2005). 

Amylose content in various short grain scented rice 

varieties ranged from 19.81-23.97 per cent. Maximum amylose 

content was observed 23.97 per cent in variety Kalanamak- 

A. The variation among the varieties is non-significant. These 

results have been close coherent from Dipti et al., 2002, 

Rayaguru, and Pandey, 2007 and Shabbir, et al., 2008.

KUMARI et. al., : Studies of quality characteristics in short grain scented rice (Oryza sativa L.) varieties/strains 179 

Total sugar content in various short grain scented rice 

varieties ranged from 1.66-1.98 per cent. Maximum total sugar 

content was observed 1.98 per cent in variety Badshah- 

Pasonda which was significantly higher over the rest varieties. 

These results are in agreement to Borua, et al., 2003. 

Reducing sugar content in various scented rice varieties 

varied from .18-.32 g/100g. Maximum reducing sugar content 

was observed in kalanamak-Berdpur (.33 g/100g) which was 

significantly superior over the rest of varieties. These results 

have been supported by Borua et al. (2003). 

Non-reducing sugar content was in ranged from 1.44- 

1.72 g/100g. Maximum non-reducing sugar content was 

observed in Badshah-Pasonda (1.72 g/100g) which was non 

significantly superior over the rest of varieties. These results 

have been favoured by Borua, et al., 2003. 


Bhattacharjee, P., Singhal, R.S. and Kulkarni, P.R., 2002. ‘Basmati 

rice: a review, International Journal of Food Science and 

Technology, 37: 1-12. 

Borua, I., Ahmed, S.A., Sarkar, C.R. and Das, D. 2003. Biochemical 

evaluation of scented rice of North India Bio-prospecting of 

commercially important plants. In: Proceeding of the national 

symposium on “Biochemical approaches for utilization and 

exploitation of commercially important plant, Jorhat, India. 12- 

14 Nov. pp. 79-85. 

Dipti, S.S.; Hossain, S.T.; Bari, S.W.; and Kabir, K.A. 2002. 

Physiochemical and cooking properties of some fine rice varieties. 

Pakistan J. Nutri., 1:188-190. 

Dubois, M., Cilles, R.A., Hamilton, J.K., Raverm, P.A. and Smith, F. 

1950. Chemistry, pp. 28-35. 

Horiuchi, H. 1966. Studies on the cereals starch (part-v) serological 

properties of the starch of rice. Agri. Biol. Chem., 30:457-465 

Juliano, B.O. 1985b. Criteria and tests for rice grain qualities. In: Rice 

Chemistry and Technology (edited by B.O. Juliano). Pp. 443–524. 

St Paul, Minnesota: American Association of Cereal Chemists. 

Juliano, B.O. 1979. The chemical basis of rice grain quality in 

proceedings of the workshop on chemical aspects of rice grain 

quality, IRRI, Los Banos, Philippines, pp. 69-90. 

Kaosa-ard, M. and Juliano B., 1992. Assessing quality characteristics 

and price of rice in selected international markets. In: Consumer 

Demand for Rice Grain Qualily, eds. (L.J. Unnevehr, B. Duft, and 

B.Juliano) International Rice Research Institute, Manila 

Mangkaeo, R., Srichuwong and Vearasilip, S. 2005. Influence of 

packaging material and storage time on seed viability and chemical 

component of rice seed, Conference on International Agricultural 

Research for Development, October, 11-13. 

Mccready, R.M., Guggolz, J., Silviera, V. and Owens, H.S. 1950. 

Determination of starch and amylose rice. Analytical Chemistry, 

22:1156-1158. 

Meullenet, J.F., Griffin, V.K., Carson, K. 2001. Rice external preference 

mapping for Asian consumers living in the United States. Journal 

of Sensory Studies, 16: 73–94. 

Miller, G.L. 1959. Use of DNS reagent for determining of reducing 

sugar. Analytical Chenistry, 31: 426-428. 

Rayagauru, K. and Pandey, J.P. 2007. Influence of extended milling on 

quality characteristics of Aromatic rice. Society for Engineering in 

Agriculture 2007. National Conference: pp. 305-312. 

Shabbir, M.A., Anjum, F.M., Zahoor, T. and Haqnawaz, 2008. Mineral 

and pasting characterization of India rice varieties with different 

milling fractions. Inter. J. Agri. Bio., 10(5):556-5650 

Siddiq, E.A., Vemireddy, L.R., and Nagaraju, J. 2012. Basmati rices: 

genetics, breeding and trade’, Agricultural Research, (1): 25-36. 

DOI 10.1007/s40003-011-0011-5. 

Singh, U.S., Singh, Neelam, Singh, H.N.; Singh, O.P. and Singh, R.K. 

2005. Rediscovering scented rice cultivar kalanamak, Asion Agri. 

History, 9(3): 211-219. 

Singh, V.P., Khush, G.S. and Dela Cruz, N. 1997. Variability and quality 

indices in aromatic rice germplasm. IRRN, pp. 22: 22. 

Trivedi, V. 2007. Biochemical compostion of aromatic rice. M.Sc. 

(Ag.) Thesis Department of Biochemistry, N.D.U.A.&T., 

Kumarganj, Faizabad. 



Studies on the Association of Plant Parasitic Nematodes Associated with Root-knot 

Nematode Infecting Potato (Solanum tuberosum) 

ZENITH N.G., JOYMATI L AND RONIBALA K.H. 

Post Graduate Department of Zoology, D. M. College of Science, Imphal 

e-mail: zenith_imp@yahoo.in, drjoymatidevi@gmail.com 

ABSTRACT 

An extensive survey was undertaken to determine the diversity 

and abundance of endemic nematodes in the potato fields of 15 

villages in Thoubal district of Manipur. The most obvious host 

response was shown by the root-knot nematode i,e Meloidogyne 

incognita in the Awang Takyenjam Leikai associated with 3 

other plant parasitic nemotodes. It was followed by the Potato 

cyst nematode Globodera spp. in Dolaithabi and Helicotylenchus 

spp. in Langmeidong area. 

Key words Diversity, nematode, Meloidogyne incognita, Potato and 

Thoubal 

Potato is one of the most important food crops in the 

world and forms the staple food for more than half of the 

population. It is popularly known as the king of vegetable 

and is very susceptible to root knot nematode (RKN). It is 

also found to be parasitized by various plant parasitic 

nematodes (PPN). In Manipur, the root knot nematode or 

Meloidogyne spp. are economically important pests of many 

crop plants, including potato. The root knot nematodes are 

soil borne and feed on roots. Their life cycle includes egg, 

juvenile and adult stage; moreover the plant parasitic 

nematodes are also soil borne pest that causes significant 

changes to agricultural and horticultural crops worldwide. So 

far, not much is known about the occurrence of plant parasitic 

nematodes associated with potato plant in this region. 

Therefore, the present study was undertaken in order to assess 

the nematode community, especially the root knot nematode 

infecting potato plant of Thoubal district. The paper highlights 

the association of 3 plant parasitic nematodes identified up to 

their generic level. 


Observations were made on the survey of root 

knot nematode in 15 different localities of Thoubal district, 

viz., Chagoning loupham, Mamang loupham, Dolaithabi, 

Khekman-mayai leikai, Waithou, Kopalli, Moijing, Keibung, 

Leisangthem-khong Manung, Leisangthem-khong Maning, 

Leisangthem –khong Mamang, Awang-Takyenjam Leikai, 

Langmeidong, Lilong chajing and Lilong haoreibi in Manipur 

from January to June, 2012. From the potato fields, the tubers 

as well as soil samples about 500gms were collected from 

around the rhizospheric regions of potato plants. Details 

relevant to the study like locality, month, date and time of 

collection were also noted. The soil samples collected in 

polythene bags were brought to the laboratory for processing, 

root samples and tubers were washed with tap water and the 

processing of the soil samples were done following the 

extraction of nematodes by Cobb’s sieving and decanting 

method followed by Baermann’s funnel method (Southey, 

1986). The nematodes collected were also processed further 

for identification and their population or counts were noted. 

Root knot species was identified as M .incognita up to their 

species level by seeing cuticular perennial pattern of gravid 

female (Norton ,1978). 


The result presented in Table 1 indicate that 3 genera of 

plant parasitic nematodes were found associated with potato 

and the root knot nematode is well distributed in a wider range 

comparing to the other plant parasitic nematodes which infect 

potato. Out of the 15 localities investigated, the root knot 

nematode has been encountered in 13 localities, except in the 

2 localities namely Kopalli and Keibung of Thoubal district. 

The highest rate of root knot nematode infection was recorded 

in Awang-Takyenjam Leikai followed by Chagoning loupham 

and medium rate of infection was seen in Leisangthem–khong 

Mamang. Globodera spp. and Helicotylenchus spp. also 

showed medium rate of infection whereas the lowest rate of 

infection was done by Tylenchus spp. The results of the 

investigation co-relates with the works of Mani and Prakash 

1992 , who reported about the association of plant parasitic 

nematodes associated with certain vegetable crops in Andhra 

Pradesh. Krishna Prasad 2006 also worked out on potato cyst 

nematodes and their management which can adjustable 

conformity with the present findings. Our results also support 

the findings of Seenivasan, et. al., 2007 investigated on 

management of potato cyst nematode through biological 

control and increased in yield production .The present 

investigation also support the findings of Joymati, et. al., 

2011 about the community analysis of plant parasitic 

nematodes in different villages of Bisnupur Districts of 

Manipur. 

The present study revealed lesser knowledge, high level 

of ignorance of the farmers of this district about plant parasitic 

nematodes associated with potato in spite of their common 

occurrence. Further, it implied that rigorous efforts are needed 

to be undertaken by all extensive agencies by using the

Table 1. 

ZENITH el. al.,: Studies on the association of plant parasitic nematodes associated with root-knot nematode infecting 181 

Association of plant parasitic nematode associated with M. incognita infecting potato plant in Thoubal diatrict of 

Manipur 

Sl. No. Localities RKN (Meloidogyne spp.) Globodera spp. Helicotylenchus spp. Tylenchus spp. 

1 Chagoning loupham +++ ++ + _ 

2 Mamang loupham + _ + _ 

3 Dolaithabi + +++ + _ 

4 Khekman-mayai leikai ++ _ _ + 

5 Waithou + ++ ++ _ 

6 Kopalli _ _ + + 

7 Moijing ++ _ _ _ 

8 Keibung _ + _ + 

9 Leisangthem-khong Manung ++ ++ + _ 

10 Leisangthem-khong Maning _ + _ _ 

11 Leisangthem –khong Mamang +++ _ + _ 

12 Awang-Takyenjam Leikai +++ ++ + + 

13 Langmeidong + _ ++ _ 

14 Lilong chajing + _ + _ 

15 Lilong haoreibi + _ _ _ 

****The symbols +++ indicates high incidence or infection by the species in the particular area, ++ indicates medium infection, + indicates lowest 

infection rate and – sign indicates the absence of the species in the areas. 

modern methods in order to disseminate the knowledge of 

these infections and their management among the farmers. 


The authors are grateful to the Head, Department of 

Zoology, D.M College of Science, Imphal for providing 

necessary laboratory facilities. They are also thankful to the 

Department of Biotechnology, Indian Institute of Science, 

Bangalore for providing financial assistance during the course 

of studies. 


Brown, E.B 1969. Assessment of the damage caused to potatoes by 

potato cyst eelworm, Heterodera rostochiensis. Annals of Applied 

Biology. 53: 493-502. 

Kaushal, K.K., Srivastava, A.N., Pankaj, Chawla, G. and Khajan Singh 

2007. Cyst forming nematodes in India – A review. Indian journal 

of Nematology 37: 1-7. 

Joymati L, Christina Kh., Jennifer O. and Bibi K. 2011. Community 

analysis of plant parasitic nematodes associated with agricultural 

crop in Bishnupur district of Manipur. Indian journal of Nematology, 

41: 220-221. 

Krishna Prasad, K.S. 2006. Potato cyst nematodes and their 

management in the Nilgiris (India). Technical bulletin No.77, Central 

Potato Research Institute (ICAR). Shimla, H.P., India, pp. 22. 

Mani, A. and Prakash, K.S. 1992. Distribution of plant parasitic 

20nematodes associated with certain principal crops in Andhra 

Pradesh. Current Nematology, 3: 21-26. 

Seenivasan, N., Devrajan, K. and Selvaraj N. 2007. Management of 

potato cyst nematodes, Globodera spp. through biological control. 

Indian journal of Nematology, 37: 21-26. 

Norton, D.C. 1978. Ecology of plant parasitic nematodes. John Wiley, 

New York. pp 266. 

Southey, J.F. 1986. Laboratory method for work with plant and soil 

nematodes, Min. Agric. Fish @ Fa., No. 402/London, HMSO. pp. 202. 



Taxonomic Study on Fishes in the Rivers of Imphal Valley 

N. MOHENDRA SINGH 

P.G. Department of Zoology Laboratory of Fishery D.M. College of Science, Imphal 795 001 

e-mail: mohendra.mangang@gmail.com 

ABSTRACT 

Altogether 53 fish specimen have been collected from different 

sapling stations of each river. The identified species belong to 

cyprinidae, Bagridae, Cobitidae, Belontidae, Channidae, 

Metacembelidae, Notopteridae, Heteropneustidae, Sisoridae, 

Belonidae, Ambassidae, Anabantidae, Claridae, Anguilla, and 

Clupeidae families. 

Key words Fishes taxonamy, Imphal 

The present study has been conducted from May 2011 

to August 2012 in the rivers of Imphal Valley. For present work 

Imphal River is selected from Chindwind River system while 

Nambul and Thoubal Rivers are the principal tributaries of 

Imphal River. 

Table 1. 

Total Nos. of fish samples in imphal river in 

different sampling stations 

Months 

Stations 

Total 

1 2 3 4 

May 3 5 - 2 10 

June 6 7 3 - 16 

July - 2 10 - 12 

August 10 2 1 8 21 

Sept. 12 - 6 7 25 

Oct. 4 5 - 2 11 

Nov. - 3 6 6 15 

Dec. 2 4 6 3 15 

Jan. 3 4 2 3 12 

Feb. 3 5 6 - 14 

March 2 1 - 10 13 

April - 10 9 6 25 

May 36 2 5 4 14 

Total 40 50 54 51 203 

Table 2. 

Total Nos. of fish samples in thoubal river in 

different sampling station in different months 

Months 

Stations 

Total 

1 2 3 4 

May 3 - 1 9 13 

June 15 36 4 6 28 

July - 5 3 3 11 

August 2 3 6 - 11 

Sept. 2 1 - 3 6 

Oct. - 4 5 - 9 

Nov. 10 - 6 4 20 

Dec. 5 2 2 - 9 

Jan. - 15 - 5 20 

Feb. - 3 6 87 16 

March 2 1 3 8 14 

April 1 3 3 7 14 

May 3 - 6 9 18 

Total 42 40 45 60 189 

Table 3. 

Total Nos. of fish samples in nambul river in 

different sampling station in different months 

Months 

Stations 

Total 

1 2 3 4 

May 2 3 - - 5 

June - 1 - 3 4 

July 3 - 2 4 9 

August - - - - - 

Sept. - 2 1 5 8 

Oct. 2 4 3 2 11 

Nov. 1 - - 6 7 

Dec. - 2 - 4 6 

Jan. - 1 - - 1 

Feb. 3 2 - 3 8 

March 1 1 - 2 4 

April - 3 - 5 8 

May - - - 2 2 

Total 12 19 6 36 73

SINGH : Taxonomic study on fishes in the rivers of imphal valley 183 

The aim of the project was to collect the fishes from the 

rivers in the valley as much as possible in 15 months. The 

other objective of the study was establish the correlation 

between the physico chemical parameter of the water and fish 

fauna in the state for academical & research purposes. 


Fishes were mostly collected from every sampling station 

randomly once in a month with the help of fisherman by using 

cast net, dip net. The collected fishes were preserved in 4-6% 

formaldehyde or10% formaline in glassware as described in 

Table 4. Check list of collected fishes in the rivers of Imphal valley 

Scientific Name Local Name Habitate 

Chitala chetala Ngapai Imphal River, Nambul River (upper region) 

Notopterus notopterus Kandla Thoubal River (Kshetri Leikai) 

Anguila bengalensis Ngaril laina Imphal River 

Gudusia chapra Wana manbi Imphal River (Koirengei) 

Labeo rohita Rou Thoubal River (Haokha) 

Labeo calbasu Ngathi Nambal River (Mantrikhong) 

Labeo gonius Kuri Imphal, Thoubal River 

Labeo pangusia Ngatin Thoubal, Imphal River 

Labeo boga Nathi Thoubal, Imphal River 

Labeo bata Ngaton Thoubal, Imphal and Nambul 

Catla catla Bao Imphal River 

Cirrhinus mrigala Mirgal Thoubal River 

Chagunius chagunio Thangol pubi Imphal River 

Tor putitora Ngara Thoubal, Imphal River 

Puntius sarana Nganoi Thoubal, Imphal, Nambul River 

Puntius sophore Phabounga Most of the study areas 

Puntius chola Phabounga Most of the study areas 

Puntius ticto Phabounga Most of the study areas 

Puntius conchonius Phabounga Most of the study areas 

Osteobrama Sps. Pengba Imphal River 

Cyprinus carpio Puklaobi Most of the study areas 

Cyprinus sps. Puklaobi Thoubal River 

Amblypharyngodon Mola Mukanga Most of the study areas 

Barilius barna Ngawa Thoubal River 

Barilius bandelisis Ngawa Thoubal River 

Barilius vagra Ngawa Imphal River 

Esomus denricus Ngasang Most of the study areas 

Rasbora rasbora Nunga Thoubal River 

Hypoathalmichthys molitrix Silver carp Nambul, Thoubal River 

Ctenopharyngodon idella Napichabi Thoubal River 

Botia dero Sarengkhoibi Most of the study areas 

Lepidocephalus guntea Ngankijou Most of the study sites 

Mystus bleekeri Ngasep Most of the study areas 

Mystus cavasus Ngasep Most of the study sites 

Wallago attu Sareng Imphal, Thoubal River 

Ompok bimaculatus Ngaten Thoubal River 

Gangata cenia Ngarang Imphal River 

Glyptothorax cavia Ngapang Imphal River 

Clarius batrachus Ngakra Nambul, Thoubal River 

Xenentodon cancila Nga Cheklaobi Thoubal River 

Chanda nama Ngamhai Most of the study areas 

Anabas testudineus Ukabi Most of the study areas 

Colisa fasciatus Ngabemba Most of the study areas 

Colisa lalia Phetin Imphal River 

Channa punctatus Ngamu bogra Nambul River 

Channa striatus Porom Nambul River 

Channa marulius Porom Thoubal River 

Macrgnathus aral Ngaril Pokchaobi Imphal River 

Mastacembelus armetus Ngaril arangba Imphal River


Shrivastava, 1968 and carried to the P.G. Fishery lab. in the 

Department of Zoology D.M. College of Science, for 

identification of fish were followed after Shrestha 1981, and 

Vishwanath (2002). 

RESULTS AND DISCUISSION 

From the present study, it was found 53 Species of fish 

and rivers in the Imphal Valley. The study would give the 

required information/data to our understanding of the river 

ecosystem in Imphal Valley which can be utilized for formation 

of suitable management policies to help conserve and 

sustainable effective productivity of this river system for fish 

and fisheries. 

ACKNOWLEDGMENT 

Author grateful for the financial assistance extended by 

UGC, (NERO), Guwahati in the form of Minor Research Project, 

I also thankful to the Director, Environment and Ecology wing, 

Manipur for helping water analysis and Head, Department of 

Zoology, D.M. College of Science, Imphal for providing 

Laboratory facilities. Thanks are also due to Dr. Kh. Rajmani 

Singh, Associate Professor, Department of Zoology (PG 

Section) D.M. College of Science for his suggestions and 

help during the study period. 


Shrestha J. 1981. Fishes of Nepal CDS, Tribhuvan University Kathmandu, 

Nepal 

Srivastava, G.J. 1986. Fishes of Eastern UP and Bihar, Rajkamal 

Prakashan, Varanashi (U.P.) India. 

Vishwanath, W. 2002. Fishes of North East India. 



Analyses of the Forest Cover Change in Rani and Garbhanga Reserve Forest, Assam, 

North East India Using Geospatial Technique 

H SUCHITRA DEVI, A. PINOKIYO 1 AND S.K. BORTHAKUR 2 

North Eastern Space Applications Centre, Govt. of India, Dept. of Space, Umiam 793 103, Meghalaya, India 

P.G. Department of Botany, D.M. College of Science, Imphal, Manipur, India 

Department of Botany, Gauhati University, Assam, India 

e-mail: suchitrahaobam@gmail.com 

ABSTRACT 

The present study was taken up for quick assessment of forest 

cover. During the study Landsat TM (1991), IRS-1D LISS-III 

sensor data (2000) and IRS P6 LISS-IV (MX) data of 2009 were 

interpreted and mapped to assess the change in the forest cover. 

The study revealed that prominent changes were observed in 

forest blank during 1991 and 2000 in Rani reserve forest. It has 

been observed that the most remarkable changes took place in 

forest blank which was 10.71% of the total area during 1991- 

2000. It was reduced to 5.65% during 1991-2009. There were 

not many changes in the forest area. However the open moist 

deciduous forest is gradually increasing during the study 

period. Similarly, in Garbhanga reserve forest prominent 

changes were observed in forest blank during 1991 and 2000 

which was 10.4% of the total area but reduced to 5.91 during 

1991-2009. Bamboo brakes were also increased during the study 

periods especially during the period 1991 and 2000. Scrub forest 

was increased only by 1.02% during 1991 and 2009. The study 

clearly showed that there has been noticeable increase in forest 

blank and reduction in medium moist deciduous forest in both 

the reserves. Built up area has also increased prominently in 

Rani reserve forest. A gradual increase in scrub forest was also 

seen in both the reserve forests. 

Key words Vegetation, deforestation, landscape. 

The rapid changes in the land utilization for various 

developmental activities from time to time necessitate a 

completely updated knowledge of the forest area and its 

changes for better management. The conventional methods 

of survey have many constraints, time consuming, costly, 

and inaccessibility to remote and difficult terrains, etc. With 

the advent of modern space technology, especially remote 

sensing, it is possible to provide synoptic view and frequent 

repetitive coverage which enables the observation of subtle 

changes. Many users world over are using remote sensing as 

an appropriate tool for data collection on land use and change 

monitoring (Green, et al., 1994, Homer, et al., 1997, Talukdar 

and Roy, 2001, Bartholome and Belward, 2005). 

The study was taken up to map the vegetation cover in 

Rani and Garbhanga reserve forest that lie close vicinity to 

Guwahati city, which has undergone rapid urban growth. The 

study presented here thus analyses the vegetation and forest 

cover changes in both the reserves so as to provide a detailed 

data on pattern of changes in forests cover for planning and 

management. 


The study was carried out for Rani and Garbhanga 

reserve forests, located in Kamrup district of Assam and the 

state of Meghalaya lies toward the southern portion of both 

the reserves. Rani reserved forest is located between 91° 35' 

16'’E to 91° 42' 24'’E Longitude and 26° 06' 41'’N to 26° 01' 

15'’N latitude, while Garbhanga reserve forest is located 

between 91° 36 25'’E to 91° 47' 45'’E longitude and 26° 05' 31'’N 

to 25° 54' 12'’N latitude (Fig. 1). 

Rani reserve forest is situated at a moderate altitude 

ranging from 60m to 401m above mean sea level and Garbhanga 

reserved forest altitude ranges from 80m to 670m above the 

mean sea level. Both the reserved forests fall on the Eastern 

part of the Kamrup district of Assam state. The rock types in 

around Rani and Garbhanga reserve forest derived form gneiss, 

quartzite or conglomerate. The soils are gravely on crests and 

upper slopes, deep red and clayey in the foot hills and alluvial 

lower down. 

Multi-date satellite data of LANDSAT-TM imagery of 

path and row no.137/42(26-11-1991), IRS ID LISS-III imagery 

of path and row no. 110/53 (07-01-2000), IRS P6 LISS-IV (MX) 

(21-08-2006) imagery were used in the study. Satellite data of 

different time period (1991,2000 and 2009) are visually 

interpreted based on the tone, texture, size, shape and pattern 

seen on imagery (Lillisand and Keifer, 1996). 

The following vegetation and land cover types were 

identified and delineated on screen using the standard visual 

image interpretation techniques. 

1. Moist deciduous forest-Closed (density >70%) 

2. Moist deciduous forest-Medium (density 40 to 70%) 

3. Moist deciduous forest-Open (density 10% to 40%) 

4. Forest- Scrub forest (Tree cover


Fig. 1. Location map of the study area 

Ground truth survey was carried out during to verify 

the interpreted areas. Land use/cover maps were prepared on 

1:25,000 (LISS-IV (MX)) and 1:50,000 (LISS-III) scales. 


Closed moist deciduous contributes the major area in 

both the reserve forest. The reserves are dominated by Shorea 

robusta that grows in association with Schima wallichii, 

Adina cordifolia, Gmelina arborea, Lagerstroemia 

parviflora, Dillenia pentagyna, Vitex peduncularis, 

Terminalia bellerica, Emblica officinalis, Premna latifolia, 

Aporosa roxburghii, Aphanamixis polystachya, Garcinia sp., 

Careya arborea, Dendrocalamus hamiltonii, Desmodium 

spp., Eupatorium odoratum, Zizyphus mauratiana, 

Microstegium ciliatum, Imperata cylindrical, Thyanolaena 

maxima, Carex stroementitia, Zizyphus oenoplia, Entada 

phaseoloides, butea parviflora, Artocarpus chaplasha, 

Michelia champaca and Amoora spectabilis , etc. Carex spp. 

occur under dense canopy. 

From the study it is found that deforestation is the main 

factor responsible for land cover change. The process of 

deforestation is illustrated in figure 2. 

It is observed that change is prominent and continuous 

from 1991 to 2009(fig 3) in the forest fringes parts surrounding 

Guwahati city due to expansion of urban area. Details of change 

in vegetation cover and land cover types are shown in 

Table 1 and Table 2. It shows the dynamics of change among 

vegetation and land cover types in Rani and Garbhanga 

reserve forest. The three sets of satellite data used for the 

monitoring the vegetation and land cover types reflected that 

changes are distinct during the period of 1991-2000 than 2000- 

2009. Further, it is evident that the blank forest contributes 

10.71 % and 10.40% during the year 1991-2000 respectively in 

Rani and Garbhanga reserve forest. In reserve bamboo brakes 

also shows an increasing trend in both the reserves. However, 

an insignificant changed was observed in closed moist 

deciduous forest. The major changes in the vegetation and 

land cover has taken place in the forest fringe area. An example 

of major deforestation i.e., complete conversion of forest to 

built up area was observed in the forest fringes (Fig. 2) in both 

the reserves. In Rani reserve forest, the most remarkable 

changes has taken place in forest blank where it was increased 

by 10.71% during 1991-2000. It has been reduced to 5.65% 

during 1991-2009. The growth in built up area changes is seen 

showing an increasing trend from 0.6 % to 34.9% during 1991- 

2000 and 2000-2009 respectively. Open moist deciduous forest 

is a gradually increasing trend during the study period. 

It was also observed during the study period in 

Garbhanga reserve forest that there were not many in closed 

and medium moist deciduous forest. However, prominent 

changes were observed in forest blank during 1991 and 2000 

which was 10.4% of the total area but reduced to 5.91 during 

1991-2009. Bamboo brakes were also increased during the study 

periods especially during the period 1991 and 2000. Scrub 

forest was increased only by 1.02% during 1991 and 2009. 

The major cause of deforestation seems to be more a societal 

need and for additional income from land. The main indirect 

stimulus for deforestation seems to be building of roads, 

transportation and communication infrastructure and 

Fig. 2. Vegetation and land cover types in 2009

Table. 1. 

DEVI et. al., : Analyses of the forest cover change in Rani and Garbhanga reserve forest, Assam, North East India 187 

Vegetation and associated land cover change in Rani reserve forest 

Categories Area in ha (1991) Area in ha 

(2000) 

Change in per 

cent 

(1991-2000) 

Area in 

ha(2009) 

Change in 

per cent 

(2000-2009) 

Change in per cent 

(1991-2009) 

Moist deciduous Forest-Closed 1799.88 1759.46 -0.02 1794.34 0.02 0.00 

Moist Deciduous Forest-Medium 1393.00 1122.00 -0.19 501.81 -0.55 -0.64 

Moist deciduous Forest-Open 766.84 991.64 0.29 1397.88 0.41 0.82 

Bamboo Brakes 131.72 154.60 0.17 195.39 0.26 0.48 

Forest-Scrub Forest 274.95 285.23 0.04 397.64 0.39 0.45 

Forest Blank 4.94 57.83 10.71 32.87 -0.43 5.65 

Agriculture land 72.49 73.00 0.01 73.03 0.00 0.01 

Built-Up area (Rural) 1.27 1.34 0.06 48.10 34.90 36.87 

Waterbodies-River - - - 4.04 - - 

Table. 2. 

Vegetation and associated land cover change in Garbhanga reserve forest 

Categories 

Area in 

ha (1991) 

Area in 

ha (2000) 

Change in 

per cent 

(1991-2000) 

Area in ha(2009) 

Change in 

per cent 

(2000-2009) 

Change in per cent 

(1991-2009) 

Moist deciduous Forest-Closed 7104.47 6545.87 -0.08 7497.74 1.23 0.06 

Moist Deciduous Forest-Medium 7369.10 4237.85 -0.42 3191.40 1.49 -0.57 

Moist deciduous Forest-Open 2499.81 3057.69 0.22 3148.94 0.85 0.26 

Bamboo Brakes 52.34 1570.57 29.01 1364.56 -0.10 25.07 

Forest-Scrub Forest 929.85 1405.20 0.51 1880.71 1.00 1.02 

Forest Blank 108.45 1235.82 10.40 748.96 -0.31 5.91 

Agriculture land 289.82 300.83 0.04 343.55 1.11 0.19 

Built-Up area (Rural) - - - 169.06 - - 

Waterbodies-River - - - 8.91 - - 

accompanying speculated land value. The Guwahati city has 

also experienced spurt in urban sprawl due to population 

growth and rural-urban migration. Highly skewed distribution 

of income and property has generated centrifugal forces 

pushing people outward from the centre to the periphery, thus 

putting more pressure on the forest. 


The authors are thankful to Director, NESAC, Shillong 

for facilities and encouragement and to the Assam Forest 

Department, Govt. of Assam for cooperation. 


Bartholome, E. and Belward, A.S., 2005, GLC 2000: a new approach to 

global land cover mapping from Earth observation data. 

International Journal of Remote Sensing Vol. 26. No. 9, 10 May 

2005, pp. 1959-1977. 

Green, K., Kempka,D. and Lackley, L. 1994. Using remote sensing to 

detect and monitor land cover and land use changes. Photogramm. 

Engg. And Remote Sensing, 60(3): 331-337. 

Lillisand, T.M. and Keifer, R.W. 1994. Remote sensing and image 

interpretation. John Wiley, New York. 

Talukdar, G. and Roy, P.S. 2001, Mapping of open Bamboo brakes and 

Forest Areas Using Satellite Remote Sensing in Meghalaya. 

Unpublished NE-SAC Report. 



Effect of Different Levels of Boron and Sulphur on Growth of Chickpea with Mustard 

Intercropping System 

SUNIL KUMAR 1 , S.K. PATEL 1 AND GAUTAM GHOSH 2 

Department of Agronomy, 2. Deptt. of Agronomy, Alld. School of Agriculture, Sam Higginbottom Institute of 

Agriculture, Technology & Sciences (Deemed-to-be-University), Allahabad - 211 007 

e-mail: sunilagro.chaudhary@gmail.com 

ABSTRACT 

A field experiment was conducted during the two consecutive 

post rainy seasons (rabi) of 2009-2010 and 2010-2011 at Crop 

Research Farm, Department of Agronomy, SHIATS, Allahabad, 

to study the effect of Boron and Sulphur on growth and yield of 

chickpea under chickpea + mustard cropping system, the 

experiment was conducted in factorial randomized block design 

with 3 replications. The combinations of treatments consisted 

of 4 intercropping system viz., sole chickpea, sole mustard, 

chickpea + mustard 4:1(row ratio) and chickpea + mustard 

5:1(row ratio) and consisted of 6 boron and sulphur levels viz., 

control, borax 10 kg ha -1 , boric acid 0.25% foliar spray at 30, 45 

and 60 DAS, sulphur 30 kg ha -1 , borax 10 kg ha -1 + sulphur 30 

kg ha -1 , boric acid 0.25% foliar spray at 30, 45 and 60 DAS + 

sulphur 30 kg ha -1 . Significantly increased plant height due to 

intercropping of chickpea with mustard 4:1 rows ratio and no. 

of branches, no. of nodules and dry weight were obtained in 

sole chickpea than intercropping of chickpea with mustard 4:1 

and 5:1 were on par. Borax 10 kg ha -1 + sulphur 30 kg ha -1 gave 

the significantly highest plant height, no. of branch, no. of 

nodules and dry weight of chickpea and mustard and it was on 

par with boric acid 0.25% trice spray + sulphur 30 kg ha -1 

followed by different borax and sulphur levels were on par over 

the control. 

Key words intercropping, boron, sulphur, growth attributes, chickpea 

and mustard 

Chickpea (Cicer arietinum L.) is a leading pulse crop in 

India, grown in 8.21 million hectares with produced of 7.48 

million tonnes during 2010 (FAOSTAT, 2012). Mustard is in 

5.77 million hectares with production of 6.59 million tonnes 

average productivity of 1142 kg ha -1 during 2009-010 (DRMR, 

2010). In India chickpea and mustard are commonly grown 

either in sole or intercropping system. Scientific approach of 

intercropping of these two crops increases the productivity 

per unit area per unit time (Ali., 1998) under a situation where 

two crops are grown in intercropping at a certain proportion 

and row ratio increase the cropping intensity, productivity 

and profitability under optimum utilization of nutrients. The 

increasing cropping intensity with higher energy requiring 

crop coupled with the use of higher used fertilizer result in to 

deficiency of boron and sulphur micronutrients, which improve 

the productivity of pulses and oil crops to a great extent. The 

important micronutrients that have been found deficient with 

major chickpea and oil crops are boron and sulphur. However, 

not much research effort has been made to augment growth 

of chickpea + mustard intercropping system. Hence, an 

experiment was conducted to study the boron and sulphur on 

growth of chickpea under chickpea + mustard cropping 

system. 


A Field experiment was conducted during post rainy 

season (rabi) of 2009-010 and 2010-011 at the Crop Research 

Farm, Department of Agronomy, SHIATS, Allahabad. The soil 

was sandy loam in texture, well drained, poor in organic carbon 

(0.34%) with pH 7.4. The available N was 114 kg, P 2 

O 5 

25 kg , 

K 2 

O 5 

223 kg ha -1 and available boron and sulphur 11.70, 0.098 

ppm, respectively. The treatments consisted of 4 intercropping 

system viz., sole chickpea, sole mustard, chickpea + mustard 

4:1(row ratio), chickpea + mustard 5:1(row ratio) and 6 boron 

and sulphur levels viz., control, borax 10 kg ha -1 , boric acid 

0.25% foliar spray, sulphur 30 kg ha -1 , borax 10 kg ha -1 + sulphur 

30 kg ha -1 , boric acid 0.25% foliar spray + sulphur 30 kg ha -1 . 

The treatment was conducted in factorial randomized block 

design with 3 replications. 

The crop was sown on 6 November in 2009 and 1 

November 2010, and harvested 10 March 2010 and 14 March 

2011. A spacing 30 X 10 cm of chickpea and 45 X 15 spacing of 

mustard, in intercropping chickpea + mustard 4:1 and 5:1 row 

ratio adapted replacement series. Fertilizers were applied as 

per the treatment, in chickpea 20 kg N ha -1 and 50 P 2 

O 5 

kg, in 

mustard sole 90 kg N ha -1 , 40 P 2 

O 5 

kg and 40 K 2 

O 5, 

in the form 

of urea, DAP and MOP. Application of boron and sulphur 

were application according to treatment basal does and foliar 

application of boric acid done at 30, 45 and 60 DAS. The test 

variety was used Pusa 362 (chickpea) and mustard (Varuna). 


Effects on chickpea : 

Intercropping systems, boron and sulphur levels was 

significant effect on plant height, no. of branches, no. of 

nodules and dry weight of chickpea (Table 1). The significantly 

highest Plant height was recorded under chickpea + mustard 

(4:1 row ratio) intercropping followed by intercropping of 

chickpea with mustard (5:1 row ratio) as compared to sole 

chickpea. Higher plant height was ascribed to shading effect 

of mustard on chickpea due to different plant architecture. 

Significantly increased no. of branches, no. of nodules and

KUMAR et al., Effect of Different Levels of Boron and Sulphur on Growth of Chickpea with Mustard Intercropping System 189 

Table1. Effect of intercropping systems, boron and sulphur levels on growth of chickpea and mustard mean of two years 

Treatments 

Plant height 

(cm) 

No. of 

branches 

Chickpea 

No. of 

nodules 

Dry weight 

(g) 

Plant height 

(cm) 

Mustard 

No. of 

branches 

Dry weight 

(g) 

Intercropping systems 

Sole chickpea 49.17 4.03 11.65 42.1 -- -- -- 

Sole mustard -- -- -- -- 163.91 5.59 77.22 

Chickpea + mustard (4:1) 52.92 3.7 8.81 32.24 161.88 5.73 79.28 

Chickpea + mustard (5:1) 51.26 3.82 9.84 35.05 159.53 5.94 81.25 

F-test S NS S S S S S 

SE (d) 0.2799 -- 0.1379 0.3859 0.371 0.0525 0.043 

CD (p=0.05) 0.5905 -- 0.291 0.8142 0.7828 0.1108 0.0906 

Boron and Sulphur Levels 

Control 49.13 3.41 8.96 34.15 157.89 5.36 78.35 

Borax 10 kg/ha 51.54 3.91 10.26 36.67 162.5 5.87 79.44 

Boric acid 0.25% spray 51.13 3.67 10.11 36.19 159.56 5.7 79.07 

Sulphur 30 kg/ha 50.44 3.53 9.61 35.44 158.76 5.63 78.74 

Borax 10 kg/ha+ sulphur 30 kg/ha 52.4 4.46 11.06 38.62 167.14 6.07 80.15 

Boric acid 0.25% spray+ sulphur 30 kg/ha 52.02 4.12 10.59 37.7 164.81 5.92 79.74 

F-test S S S S S S S 

SE (d) 0.3958 0.1954 0.1951 0.5457 0.5247 0.0742 0.0608 

CD (p=0.05) 0.8351 0.4123 0.4116 1.1514 1.107 0.1566 0.1282 

dry weight were recorded in sole chickpea than intercropping 

of chickpea with mustard 5:1 row ratio. However, intercropping 

of chickpea with mustard 4:1 row ratio recorded the minimum 

of these parameters this could be attributed to more space 

available to individual plant for lateral spread. 

Boron and sulphur levels gave the significantly 

increased plant height, no. of branches, no. of nodules and 

dry weight compared to control. Significantly the highest of 

this parameters was obtained from application of borax 10 kg 

ha -1 + sulphur 30 kg ha -1 than application of boric acid 0.25% 

spray + sulphur 30 kg ha -1 but it was statistically equivalent to 

borax 10 kg ha -1 and sulphur 30 kg ha -1 , respectively. It might 

be ascribed to addition of boron and sulphur is attributed to 

their role in carbohydrates metabolism, protein synthesis and 

activation of number of enzymes in the soil which deficiencies 

occur these nutrients and increased growth components of 

chickpea. These results are in conformity with those of Meena, 

et.al., 2005. 

Effects on mustard : 

Intercropping systems, boron and sulphur levels had 

significant effect on plant height, no. of branches and dry 

weight of mustard (Table 1). Sole mustard gave the significantly 

highest plant height than mustard intercropped in chickpea 

(4:1 row ratio), but no. of branches and dry weight was found 

highest in mustard intercropped in chickpea (5:1 row ratio) 

followed by mustard intercropped in chickpea (4:1 row ratio). 

This may be owing to more plant per unit area and less interaspace 

competition posed by sole mustard and plant height 

decreased by different row proportion through wide space of 

mustard with chickpea due to different plant architecture. 

Among the boron and sulphur levels gave the 

significantly increased plant height, no. of branches and dry 

weight over the control. Application of borax 10 kg ha -1 + 

sulphur 30 kg ha -1 gave the significantly highest plant height, 

no of branches and dry weight, followed by boric acid 0.25% 

spray + sulphur 30 kg ha -1 but it was statistically on par with 

borax 10 kg ha -1 and boric acid 0.25% spray, respectively. This 

may be probably due to adequate application of boron and 

sulphur which were directly involved in better absorption of 

applied nutrients and cell multiplication as well as expansion 

of deep green color of leaves due to better chlorophyll 

synthesis in comparison with plants deficient in boron and 

sulphur. These results were confirmed by Kumar et al. (2006). 


Ali, M. 1998. Chickpea-based intercrops for command areas of different 

agro-ecological zones of India. In: Proceeding of national 

symposium on efficient cropping system zone of India held during 

of 7-10 January at University of Agricultural Science, Bangalore, 

pp. 53. 

D.R.M.R. 2010. Directorate of Rapeseed-Mustard Bharatpur 

(Rajasthan), India. (http://www.drmr.res.in). 

FAOSTAT. 2012. Food and Agriculture Organization of the United 

Nations, Rome, Italy; Crop Production Database (http://fao.org). 

Meena, S.K., Sharma, S. and Meena, H.S. 2005. Effect of sulphur and 

zinc fertilization on yield, quality and nutrient content and uptake 

of chickpea (Cicer arietinum L.) under semi arid tropics. Ann. 

Agric. Res. News Series, 26 (1): 45-47. 

Kumar, A., Prasad, S. and Kumar, S.B. 2006. Effect of boron and 

sulphur on performance of gram (Cicer arietinum). Indian J. Agron., 

51 (1): 57-59. 



Efficacy of Fungicides on In Vitro Growth of Pigeonpea against Stem Canker 

1 

SRUJANI BEHERA, 2 R. B. SINGH AND 2 LAXMAN PRASAD BALAI 

1 

Department of Plant Pathology, Bidhan Chandra Krishi Viswavidyalaya Mohanpur, Nadia, 

West Bengal - 741 252, 

2 

Mycology and Plant Pathology, Institute of Agriculture Sciences, Banaras Hindu University, Varanasi 221005 

e-mail : srujani.lucy@gmail.com 

ABSTRACT 

Phoma stem canker of pigeonpea caused by Phoma cajani is a 

sporadic disease but occasionally attains highly destructive 

proposition and becomes epidemic highly conducive 

environmental condition. Four fungicides viz., SAAF 

(Carbendazim 12% + Mancozeb 63%), Benfil (Carbendazim 

50%), Vitavax (Carboxine 75%), and Dithane M-45 (Mancozeb 

75%) were evaluated against Phoma cajani under in vitro 

condition by poison food technique. The results indicated that 

SAAF was found to be the most effective in suppressing the 

growth followed by Benfil. Among the different concentrations 

tested, SAAF at 350 ppm and Benfil at 400 ppm were found to 

be optimum for the control of pathogen’s growth. 

Key words Phoma stem canker, poison food technique. 

Phoma stem canker of pigeonpea (Cajanus cajan (L) 

Millsp.) incited by Phoma cajani, when it was considered to 

be a minor disease previously but become progressively severe 

in past years causing more economic losses to pigeonpea 

production in recent years. The disease was repeatedly seen 

in experimental plots and farmers fields causing 5-50 % 

mortality in plants at maturity stage during periodical surveys 

and critical observations. During the year 2009-2010 pigeonpea 

has a role in pulse production in India. The stem canker disease 

is a great obstacle in the way pivot able and presently 

management of this devastating disease is of prime importance. 

In the present study a few fungicides were screened for their 

fungitoxic activity against P. cajani. An in vitro effect on 

colony diameter was initially investigated. In vitro screening 

of fungicides has been employed by many plant pathologists 

(Mostert, et al., 2000; Ponmurugan, et al., 2006; Girishab, 

et. al., 2009). 


Poison food technique (Dhingra and Sinclair, 1995) was 

adopted to elucidate the efficacy of different fungicide viz. 

SAAF (Carbendazim 12% W.P. + Mancozeb 63% W.P.), Benfil 

(Carbendazim 50% W.P), Vitavax (Carboxine75% W.P.) and 

Dithane M-45 (Mancozeb 75% W.P.). Required amount of 

different fungicides were added to 250 ml Erlenmeyer flasks 

containing 100 ml of melted PDA medium to make 50, 100, 150, 

200, 250, 300, 350, 400, 450, 500 ppm concentrations, 

respectively. The medium of each flask was poured into Petri 

dishes and allowed to solidify. Mycelium disc (5mm dia.) were 

cut with the help of cork borer from the growing edge of 6-7 

days old culture of the fungus grown on PDA and transferred 

in to the centre of each petridishes. The petridishes containing 

medium without fungicides served as control. The experiment 

was repeated with four replications and the petridishes were 

incubated at 25 0 ± 2 0 C temperature. Radial growth of the fungus 

was measured at an interval of 24 hrs till it reached the edge of 

the petri plate (Dhingra and Sinclair, 1995; Nene and Thapliyal, 

2001). Relative growth of fungus at each concentration was 

evaluated by comparing it with the control for each fungusfungicide 

combination. The per cent inhibition (PI) of the 

fungus over control was calculated using the (Ponmurugan, 

et al., 2006) following formula: 

PI = 

(A - B) 

×100 

A 

Where, A is colony diameter of the fungus in control 

plates (mm) 

B is colony diameter of the fungus in treated plates (mm). 


Response of Phoma cajani to ten concentrations (i.e., 

50, 100, 150, 200, 250, 300, 350, 400, 450 and 500 ppm) of different 

fungicides viz., SAAF, Benfil, Vitavax and Dithane M 45, was 

studied to determine differences in tolerance of the fungicide 

to various toxicants. Table 1. elucidates that different 

fungicides have profound inhibitory effect on isolate of 

P. cajani at its different concentrations. There was significant 

difference among the mycelial growth of P. cajani obtained at 

different dosages of fungicides. Pronounced reduction of 

growth occurred in medium with increasing concentrations of 

fungicides. Maximum per cent inhibition of radial growth of 

the fungus was observed at a concentration of 300 ppm 

(83.33%) of SAAF, 350 ppm (80%) of Benfil, respectively. 

However Vitavax and Dithane M 45 were required at equal 

concentrations (400 ppm) for maximum per cent inhibition of 

radial growth (86.38%, 81.94% respectively) of the fungus. 

The growth of stem containing pathogen was inhibited 

completely at one or another concentration. SAAF most 

effectively controlled P. cajani at 350 ppm. In contrast, Benfil, 

Vitavax and Dithane M 45 required comparatively higher 

concentrations (400 ppm, 450 ppm and 450 ppm respectively) 

than SAAF for complete inhibition of mycelial growth.

Table 1. 

MURALI : Nematode Infecting Thrips and Their Utilization In Pest Management: A Review 191 

Effect of different fungicides on radial growth of P. cajani 

Fungicides 

Per cent inhibition of radial growth* 

Concentration (ppm) 

50 100 150 200 250 300 350 400 450 500 Mean 

SAAF 40.27 48.61 55.55 71.38 78.04 83.33 100.00 100.00 100.00 100.00 77.72 

Benfil 30.83 40.83 49.44 60.83 69.16 74.44 80.00 100 100.00 100.00 70.55 

Vitavax 16.93 29.16 39.16 54.44 63.88 73.88 78.88 86.38 100.00 100.00 64.27 

Dithane M 45 14.44 25.27 31.38 39.99 50.27 62.22 71.94 81.94 100.00 100.00 57.74 

SEm± 1.92 1.85 1.73 1.34 2.54 1.26 0.92 1.00 0.00 0.00 0.00 

CD (P=0.05) 4.2 4.04 3.78 2.93 5.53 2.74 2.02 2.18 0 0 0 

CD (P=0.01) 5.89 5.67 5.3 4.12 7.76 3.85 2.83 3.06 0 0 0 

*On 5th day after inoculation 

In present studies SAAF, Benfil, Vitavax and Dithane 

M-45 completely suppressed radial growth of P. cajani at a 

particular concentration. Significant decrease in mycelial 

growth with higher concentration of different fungicides was 

recorded. Among the 4 chemical fungicides used, SAAF 

(Carbendazim 12% + Mancozeb 63%) was found to be most 

effective at a comparatively lower concentration (300 ppm) 

followed by Benfil (Carbendazim 50%) whereas Vitavax and 

Dithane M 45 required equal concentrations (400 ppm) for 

maximum percent inhibition of radial growth of the fungus. 

In present study, Benfil (Carbendazim 50%) was more 

effective than Dithane M 45. These in formations corroborate 

the findings of Potdukhe, 1998. Out of six fungicides tested, 

he found carbendazim (0.1 and 0.25%) was best in vitro 

followed by Dithane M 45 (0.4 and 0.25%) for inhibiting the 

growth of pathogen. 

In previous observations amongst the chemicals tested, 

Calixin was most effective followed by Dithane M 45 (Somani 

and Rout, 988) but in the present investigation instead of 

Calixin, Vitavax was taken and it was deduced that Vitavax and 

Dithane M-45 have same effect at one concentration but 

Vitavax (86.38%) had advanced radial growth than Dithane M 

45(81.94%). 

After computing it is concluded that is our findings the 

commercial fungicide i.e., SAAF (Carbendazim 12% + 

Mancozeb 63%), which was a mixture of both systemic and 

contact fungicides was best among all the chemicals used. 

There is no earlier findings regarding this hence, further 

research is needed to confirm the aforsaid previous results. 


Dhingra, O.D. and Sinclair, J.B. 1995. Basic plant pathology methods. 

2nd ed. Boca Raton, FL: CRC Press. pp. 267-285. 

Girishab, K., Shankara, B.S. and Raveesha, K. A. 2009. In vitro screening 

of systemic fungicides against Phomopsis azadirachtae, the incitant 

of die-back of neem. Archives of Phytopathology and Plant 

Protection, 42(3): 256–264. 

Mostert, L., Denman, S and Crous, P.W. 2000. In vitro screening of 

fungicides against Phomopsis viticola and Diaporthe perjuncta. 

S. Afr. J. Enol. Vitic., 21: 62 – 65. 

Nene, Y.L and Thapliyal, P.N. 2001. Fungicides in Plant Disease 

Control. New Delhi: Oxford and IBH Publications. 

Ponmurugan, P, Baby, U.I. and Gopi, C. 2006. Efficacy of certain 

fungicides against Phomopsis theae under in vitro conditions. Afr. J. 

Biotechnol., 5: 434 – 436. 

Potdukhe, P.A. 1998. Fungal diseases of root and stem. Published in the 

book. Diseases of pigeonpea. pp. 72-77. 

Somani, R.B. and Raut, B.T. 1988. Efficacy of different fungicides III. 

Pigeonpea stem canker. PKV Research Journal, 12(2):, 171-173. 

Received on 20.7.2012 August 28.09.2012


Influence of Microbial Inoculants and Nutrients on Morpho-Physiological, Growth 

Parameters and Yield Potential in Tomato (Lycopersicon esculentum L. Mill.) 

MOHAN KUMAR, K.G., AND CHETTI M.B. 

Department of Crop Physiology, University of Agricultural Sciences, Dharwad, 

e-mail: mohankumar.kg@rediffmail.com 

ABSTRACT 

A field experiment was conducted during kharif, 2002 at Main 

Agricultural Research Station, University of Agricultural 

Sciences, Dharwad, to study the influence of microbial 

inoculants and nutrients on morpho- physiological traits and 

yield potential in tomato cultivar Mega (L. 15). The experiments 

were laid out in randomized block design with three 

replications. The experiment consisted of three levels of 

nitrogen (30, 45 and 60 kg/ha), three levels of phosphorus 

(25,37.5 and 50 kg/ha) and inoculation with Azospirillum 

brasilense, Pseudomonas striata individually and in combination 

to seed, and seedling . The results reveled that the treatment 

received RDF along with both inoculants recorded significantly 

higher values for morpho-physiological, yield and yield 

components. 

Key words microbial inoculants, Azospirillium, Pseudomonas, yield. 

Tomato is fertilizer responsive crop, the frequent 

application of chemical fertilizers leads to environmental 

hazards especially nitrate, which is major threat causing 

serious health hazards viz., methaemoglobinaemia ,stomach 

cancer etc.and also affecting natural waters by 

eutrophication(Hester and Harrison.1996). Therefore the 

current trend is to explore the possibility of supplementing 

chemical fertilizers with biofertilizers, more particularly of 

microbial origin along with organics. Microbial processes are 

not only quick but also consume relatively less energy than 

industrial processes, secondly they have the advantage of 

being diversified into small units to meet the demands of the 

specific problems of location which is apt to come across in 

the agricultural practice of nations which are not mechanized 

in farming (Subba Rao,1998).Therefore the present 

investigation was under taken to study the effect of microbial 

inoculants and nutrients on morpho- physiological traits and 

yield potential in tomato. 


The experiment was conducted during kharif,2002 at 

College of Agriculture, University of Agricultural Sciences, 

Dharwad. The experiment consisted of fourteen treatments 

(Table 1). 

RDN = Recommended dose of nitrogen, RDP = 

Recommended dose of phosphorus, RDK = Recommended 

dose of potassium 

The experiment was laid out in randomized block design 

with three replication. The seeds of variety Mega (L. 15) were 

treated with captan and were inoculated separately with 

Azospirillum brasilense, Pseudomonas striata and mixture 

of Azospirillum brasilense and Pseudomonas striata. After 

four weeks healthy and uniform seedlings were selected for 

transplanting. Seed inoculated seedlings were dipped in 

respective biofertilizer slurry for 30 minutes. Inoculated and 

un inoculated seedlings were transplanted to the main field at 

spacing of 75 cm x 60 cm. Five plants were selected randomly 

and tagged in each plot for recording various observation at 

different stages. 


The data with respective to morpho-physiological traits 

viz., plant height, number of branches per plant, number of 

leaves per branch and leaf area showed significant differences 

due to microbial inoculants and nutrients at all stages. Among 

the treatments RDF + Azospirillum brasilense + Pseudomonas 

striata recorded significantly higher values for these traits at 

all stages. The growth parameters such as Leaf area index, 

leaf area duration, total dry matter, crop growth rate, absolute 

growth rate, net assimilation rate relative growth rate were 

differed significantly within the treatments, maximum values 

for all these parameters were recorded in the treatment which 

received RDF + Azospirillum brasilense + Pseudomonas striata 

(Table 2). 

The data on yield and yield components presented in 

Table 2 indicated significant differences between treatments 

with respect to all the paramters viz., 100 – seed weight, number 

of fruits per plant, fresh weight per fruits, fruit yield per plant 

and fruit yield per hectare. Among the treatments RDF + 

Azospirillum brasilense + Pseudomonas striata recorded 

significantly higher values for these components. The 

significant differences were observed due to dual inoculation 

of Azospirillum brasilense and Pseudomonas striata with 

RDF in all morpho-physiological traits studied. The increase 

in plant height at all the stages might be attributed to N2- 

fixation by Azospirillum and P – solubilization by 

Pseudomonas striata which in turn make these two essential 

nutrients available to the plant growth and development. 

These substances have also been reported to increase the 

activity of cell division and cell elongation ultimately leading 

to increased plant height. Similar results have also been

KUMAR et. al., : Influence of Microbial Inoculants and Nutrients on Morpho-Physiological, Growth Parameters 193 

Table 1. Influence of microbial inoculants and nutrients on growth parameters in tomato 

Days after transplanting 

Treatments 

Total dry matter 

Crop growth rate Absolute growth rate 

(g/plant) 

(CGR, g/m2/day) 

(AGR, g/day) 

30 60 90 At harvest 30-60 60-90 90-harvest 30-60 60-90 90-harvest 

T1 – Recommended dose of fertilizer 14.2 47.9 86.1 139.6 2.61 2.71 3.90 1.17 1.22 1.75 

T2 – 50%RDN + RDP + RDK 10.0 34.0 69.9 110.8 1.78 2.41 3.02 0.80 1.09 1.36 

T3 – 75% RDN +RDP + RDK 11.8 38.7 73.9 118.3 1.98 2.56 3.29 0.89 1.18 1.49 

T4 – 50% RDN +RDP + RDK + Azo. 12.7 43.4 76.1 124.6 2.27 2.63 3.50 1.02 1.19 1.57 

T5 – 75% RDN +RDP + RDK + Azo. 13.0 47.0 83.1 135.4 2.43 2.68 3.87 1.09 1.20 1.74 

T6 – RDF + Azo. 16.4 58.4 97.4 153.0 23.11 2.86 4.12 1.40 1.29 1.86 

T7 – RDN + 50% RDP + RDK 11.8 38.3 73.2 117.7 1.97 2.55 3.29 0.89 1.15 1.48 

T8 – RDN + 75% RDP + RDK 12.0 41.5 76.0 123.2 2.20 2.62 3.46 0.99 1.18 1.56 

T9 – RDN +50%RDP + RDK + Pseu. 12.7 43.7 79.1 126.0 2.27 2.63 3.59 1.02 1.19 1.62 

T10 – RDN +75%RDP + RDK +Pseu. 13.0 44.5 80.1 131.6 2.34 2.64 3.82 1.05 1.20 1.72 

T11– RDF + Pseu. 14.5 54.3 89.9 146.3 2.89 2.83 4.11 1.30 1.28 1.85 

T12 – 50%RDN +50%RDP + RDK+ Azo.+ Pseu. 15.3 51.0 90.8 142.5 2.71 2.78 3.95 1.22 1.25 1.78 

T13 – 75%RDN +75%RDP + RDK + Azo. + Pseu 18.0 62.2 99.7 157.1 3.28 2.89 4.24 1.47 1.30 1.91 

T14 – RDF + Azo. + Pseu. 19.4 64.9 103.5 160.8 3.37 2.89 4.25 1.52 1.30 1.91 

Mean 13.90 47.80 84.20 134.80 2.52 2.69 3.74 1.13 1.22 1.69 

SEm ± 0.83 2.41 3.24 4.78 0.05 0.33 0.41 0.08 0.19 1.19 

CD at 5% 2.42 6.99 9.41 13.90 0.18 NS NS 0.24 NS NS 

Table 2. Influence of microbial inoculants and nutrients on growth, yield and yield components in tomato 

Treatments 30-60 

DAT 

Net assimilation rate 

(NAR, g/m2/day X 10-2) 

60-90 

DAT 

90- 

harvest 

Relative growth rate 

(RGR, g/g/day) 

30-60 

DAT 

60-90 

DAT 

90 - 

harvest 

100-seed 

weight 

(mg) 

Yield and yield components 

No. of 

fruits per 

plant 

Fresh 

weight 

(g/fruit) 

Fruit 

Yield 

(g/plant) 

T1 – Recommended dose of fertilizer 0.916 0.369 0.56 0.042 0.02 0.016 324.0 22.0 50.0 891.7 19.83 

T2 – 50%RDN + RDP + RDK 0.683 0.3 0.442 0.039 0.016 0.01 282.0 18.4 38.0 681.0 15.13 

T3 – 75% RDN +RDP + RDK 0.72 0.332 0.517 0.04 0.017 0.017 298.0 19.7 42.0 790.8 16.61 

T4 – 50% RDN +RDP + RDK + Azo. 0.81 0.346 0.526 0.04 0.019 0.015 307.0 20.0 45.0 801.5 17.80 

T5 – 75% RDN +RDP + RDK + Azo. 0.841 0.356 0.557 0.041 0.02 0.016 322.0 21.5 48.0 889.4 19.75 

T6 – RDF + Azo. 1.003 0.379 0.584 0.042 0.021 0.016 336.0 22.4 55.0 1120.1 24.88 

T7 – RDN + 50% RDP + RDK 0.713 0.33 0.489 0.039 0.016 0.015 295.0 19.1 40.0 780.6 17.34 

T8 – RDN + 75% RDP + RDK 0.83 0.34 0.525 0.04 0.017 0.015 305.0 19.8 43.0 793.2 17.66 

T9 – RDN +50%RDP + RDK + Pseu. 0.813 0.354 0.531 0.041 0.019 0.015 312.0 20.2 45.3 837.8 18.61 

T10 – RDN +75%RDP + RDK +Pseu. 0.83 0.355 0.532 0.041 0.019 0.016 318.0 20.2 47.7 862.9 19.17 

T11– RDF + Pseu. 0.96 0.377 0.58 0.042 0.02 0.016 333.0 22.3 51.3 1006.2 22.35 

T12 – 50%RDN +50%RDP + RDK+ Azo.+ Pseu. 0.915 0.37 0.572 0.042 0.02 0.016 328.0 22.2 51.0 898.7 19.92 

T13 – 75%RDN +75%RDP + RDK + Azo. + Pseu 1.055 0.385 0.588 0.042 0.022 0.017 338.0 24.7 60.0 1169.5 24.88 

T14 – RDF + Azo. + Pseu. 1.066 0.426 0.592 0.044 0.024 0.018 352.0 26.0 64.0 1280.9 28.46 

Mean 0.866 0.358 0.542 0.041 0.019 0.015 317.93 21.30 48.60 914.60 20.17 

SEm ± 0.067 0.048 0.6 0.003 0.002 0.002 10.36 1.18 2.03 37.99 0.83 

CD at 5% 0.196 NS NS NS NS NS 30.11 3.42 5.89 110.37 2.43 

Fruit 

Yield 

(t/ha) 

reported by Fallik and Okon, 1996 in Setaria italica and coffee 

and tea seedlings Merina, 1995. The increased number of 

branches and number of leaves could be because of certain 

growth promoting substances secreted by microbial 

inoculants and the availability of more nitrogen and 

phosphorus which in turn lead to better root development, 

better translocation of water uptake and deposition of 

nutrients. 

It is well established that the infrastructure of the plant 

is decided by the growth parameters like leaf area, LAI, LAD, 

TDM, CGR,RGR,NAR and AGR. In the present investigation, 

it was observed that the treatments differed significantly with 

respect to LA,LAI,LAD and TDM at all the growth stages 

and not differed significantly with respect to CGR,AGR and 

NAR at later stages and RGR at all growth stages due to dual 

inoculation of Azospirillum brasilense and Pseudomonas 

striata with RDF. The increase LA and LAI could be attributed 

to increased cell division and cell elongation resulting in 

increased leaf expansion, more number of leaves and branches 

per plant due to beneficial influence of biofertilizers which 

release growth promoting substances and enhance the 

availability of both nitrogen and phosphorus.


More LAD might be attributed to the greenness due to 

dual inoculation of Azospirillum and Pseudomonas with RDF. 

The increased TDM in the plants which received both the 

inoculants and the RDF could be attributed to their influence 

on leaf area, LAI, CGR, AGR, RGR and NAR. In the present 

investigation the fruit yield per plant and per hectare were 

significantly higher in the treatment of RDF with dual 

inoculation of Azospirillum and pseddomonas. This might be 

due to the production of more lateral roots, increased symbiotic 

activity of nitrogen fixation and P-solubilization which have 

helped in getting higher nitrogen and phosphorus required 

for growth and development. In the present investigation it is 

seen from the results that there was a significant increase in 

all the morphological, growth, yield and yield components 

due to RDF with dual inoculation of Azospirillum and 

Psedomonas thus signifying the role of biofertilisers. 

LITERATRURE CITED 

Fallik and Okon, Y., 1996, Inoculation effect of Azospirillum brasilense 

on biomass production, survival and growth promotion to setaria 

italica and Zea mays. Soil Biology and Biochemistry, 128: 

123-126. 

Hester, R.E. and Harrison, R.M., 1996, Agriculture chemicals and 

environment, pp. 1. 

Merina, P.S., 1995, Response of certain horticultural crops to inoculation 

with fungi azospirrilum and phosophobacteria. M.Sc.(Agri.) Thesis. 

Tamil Nadu Agricultural university, Coimbatore. 

Subbarao, N.S., 1998, Nitrogen fixing bacteria associated with plantation 

and orchid plants. Canadian Journal of Microbiology, 29: 863-866. 



Sensory Quality Evaluation of MA Packaged Fruits Applying Fuzzy Logic 

S.MANGARAJ 1 , M.K.TRIPATHI 2 

Agro Produce Processing Division, Central Institute of Agriculture Engineering, Nabi Bagh, Baresia Road, 

Bhopal, M.P. India 

e-mail: sukhdev0108@gmail.com 

ABSTRACT 

The laminated MA packed apple, guava and litchi were taken 

after specific storage periods and kept at the recommended 

ripening temperatures and RH for determining the sensory 

qualities like color, texture, aroma and taste. Eleven judges 

were selected based on good health, interests and knowledge 

in sensory evaluation, ability to concentrate, learned and 

familiarity with the fruits. They were asked to judge fruit 

samples quickly but not in hurry. Sensory scale factors assigned 

to each of the quality attributes (viz., color, texture, aroma and 

taste) were not satisfactory, fair, medium, good and excellent. 

The panelists were asked to rank the fruit samples by giving 

tick marks to appropriate scale factor for each of the quality 

attributes. The data obtained from the judges were processed 

applying Fuzzy logic modeling using MATLAB programme. 

The results indicated that the similarity in values under very 

good and good category is the highest for MA packaged apple, 

guava and litchi. Hence the overall quality of these samples 

were ranked first / best as compared to the control storage 

fruits. The important / order of preference of quality attributes 

for apple, guava and litchi fruits, in general, was rated as taste 

> aroma > color > mouth feel; taste > mouth feel > aroma > 

color; and colour > aroma > taste > mouth feel, respectively. 

Key words MA packaging, fuzzy logic, modelling, sensory evaluation 

Sensory evaluation is the science of judging and 

evaluating the quality of a food by the use of the senses, i.e. 

taste, smell, sight, touch and hearing (Das, 2005; Falade and 

Omojola, 2008). Fuzzy logic is an important tool by which 

vague and imprecise data can be analyzed and important 

conclusions regarding acceptance, rejection, ranking, strong 

and weak attributes of food can be drawn. 

MA packaging is a food packaging method in which the 

proportion of CO 2 

, O 2 

and N 2 

in a sealed container are different 

from those in the normal air to enhance the food shelf-life 

(Kader, et. al., 1989, Mangaraj, et, al., 2011). It involves the 

exposure of produce to the atmosphere generated in a package 

by the interaction of the produce, the package and the external 

atmosphere (Mangaraj and Goswami, 2008; Mangaraj et al., 

2009; Montanez, et al., 2010). The higher CO 2 

and lower O 2 

atmosphere surrounding the commodity, potentially reduce 

respiration rate, ethylene sensitivity, and production, 

physiological changes and decay (Mahajan, et al., 2007; 

Mangaraj and Goswami, 2009a). The fruits apple (cv. Royal 

Delicious), guava (cv. Baruipur) and Litchi (cv. Shahi) 

harvested from the orchard at their commercial maturity 

(Mangaraj and Goswami, 2009d), were sealed in laminated MA 

packages and kept for storage study at different temperatures. 

Though the quality attributes were determined objectively at 

regular intervals, various sensory attributes such as color, 

texture, taste and mouth feel of fruit samples were evaluated 

and Fuzzy logic model was developed for the sensory 

evaluation of stored fruits (Mangaraj, et. al., 2005; Mangaraj, 

et. al., 2006). The main objective of this study was to evaluate 

the sensory scores of different MA packaged and unpackaged 

fruit samples using fuzzy logic and grade the samples as per 

their sensory qualities to find out the strength and weakness 

of individual sample, preference of quality attributes of fruits 

in general and ranking of fruits. 


Raw materials : 

The fruits apple (cv. Royal Delicious), guava (cv. 

Baruipur) and Litchi (cv. Shahi) were harvested from the 

orchard at their commercial maturity. It was ensured to maintain 

uniformity in terms of size and weight of individual fruits in 

the whole lot of samples. 

Selection of polymeric films : 

With the objective of meeting MAP requirements the 

polymeric films namely LDPE, BOPP, PVC, PVDC were 

procured considering various film characteristics such as gas 

transmission rates for O 2 

and CO 2 

, WVTR, clarity, strength, 

printability and cost effectiveness (Exama, et al., 1993; Costa, 

et al., 2011). The gas transmission rates of films were measured 

using standard methods (Mangaraj, et al., 2012 a and b). 

Development of MA packages : 

The different combination of PVC and BOPP as well as 

that of PVC and LDPE was tailored for lamination to bring the 

gas transmission characteristics of the laminates close to the 

required values. Using these combinations five types of film 

laminates for MA packages viz., PCG-LFR-1 and PCG-LFR-2 

for apple; PCG-LFR-3 and PCG-LFR-4 for guava; and PCG- 

LFR-5 for litchi were developed. A package size of 24 cm x 19 

cm, 19 cm x 19 cm, and 28 cm x 22 cm for a fill weight of 1.00 kg 

± 100 g was found to be appropriate for MA packaging apples, 

guava and litchi (Mangaraj, et. al., 2012a,b, 2013)


Evaluation of MA packages : 

The MA packages were labeled marked and kept in the 

incubator at 10, 15, 20 and 25 ºC for storage study. The 

performance of various packages was evaluated for their, 

ability to extend the shelf life of the packaged fruit. The different 

quality parameters and sensory attributes such as color, 

texture, taste and mouth feel of fruit samples were evaluated 

at regular interval during storage. 

Fuzzy logic model for the sensory evaluation fruits : 

In Fuzzy modelling, linguistic variables (not satisfactory, 

good, excellent etc.) were used for developing relationship 

between independent (color, aroma, taste, mouth feel) and 

dependent variables (acceptance, rejection, ranking, strong 

and weak attributes of food). This modelling utilized the 

linguistic data from the subjective evaluation along with the 

accurate and precise data variable from objective evaluation. 

Sensory analysis : 

For sensory evaluation, fruit samples were taken out 

from MAP and control storage and held at room temperature 

for ripening. The fruit samples was placed on white plates and 

presented to a taste panel of 11 judges familiar with the quality 

and sensory parameters of fruits. Each sample was identified 

by a random two-digit code. The order of presentation of the 

samples on the plates was randomized for each panelist. The 

quality attributes selected for characterizations of fruits were: 

colour, texture, aroma and mouth feel. Judges were asked to 

give tick mark to appropriate scale factor for each of the quality 

attributes as well as the to give rank to quality attributes of 

fruits in general. The panelists assessed all the samples and 

at the same time gave tick mark/score in the score sheet to the 

above-mentioned parameters. 

Fuzzy logic modelling for the sensory evaluation of 

fruits : 

Triplets associated with sensory scales : 

Considering triangular fuzzy membership distribution 

function for all the sensory scale factors, overall ranking of 

four fruit samples, most and least important quality attributes 

of fruits in general, and strong and weak quality attributes of 

sample was determined. Triangular membership pattern of 

b 

1 

a 

0 

Not satisfactory / 

Not at all 

important 

Fig. 1. 

c 1 

c 

25 

Fair / 

Somewhat 

important 

d 1 

d 

50 

Medium / 

important 

75 

Good / highly 

important 

1 

Value of fuzzy 

membership 

function 

100 

Excellent / 

Extremely 

important 

Values of Triplicates Associated with Triangular 

Membership Distribution Function 

sensory scale was represented by a set of three numbers, 

called as triplet, which was formed as triangle a b c representing 

membership distribution function for not satisfactory / not at 

all important category, triangle a c 1 

d representing distribution 

function for fair/somewhat important category (Fig. 1). 

Table-1 showed the ‘triplets’ associated with five point 

sensory scale. 

Table 1. 

Not satisfactory/ 

not at all 

important 

Triplets associated with sensory scales 

Fair / 

somewhat 

important 

Medium / 

important 

Good / 

highly 

important 

Excellent / 

extremely 

important 

0 0 25 25 25 25 50 25 25 75 25 25 100 25 0 

Triplets for sensory score of fruits : 

For a particular sample, a quality attribute, and sensory 

scores; a triplet was obtained from of sensory scores, triplets 

associated with the sensory scale and no. of judges. For 

example, for PCG-LFR-1 sample and its taste attributes, value 

of triplet was found out using equation (1) as follows. In similar 

manner, values of triplets for color, aroma and mouth feel for 

PCG-LFR-1 sample was calculated. 

N 

j 0 0 25 + N 

j 25 25 25 + N 

j 50 25 25 + N 

j 75 25 25 + N 

j 100 25 0 

PCG-LFR-1T = 

TN 

j 

…(1) 

Where, N j 

is the no. of judges associated with the triplets 

and TN j 

is the total numbers of judges used for sensory 

evaluation 

Triplets for sensory score of quality attribute : 

For a particular quality attribute fruits sample in general 

and its sensory scores; the triplet was obtained from sum of 

sensory scores, triplets associated with the sensory scale 

and number of judges. For the quality attributes taste, the 

value of triplets QT was obtained employing equation (2). 

The values of triplets for color, aroma and mouth feel of fruits 

sample was calculated similarly. 

…(2) 

N 

j 

0 0 25 + N 

j 

25 25 25 + N 

j 

50 25 25 + N 

j 

75 25 25 + N 

j 

100 25 0 

QT = 

TN 

Triplets for relative weightage of quality attributes : 

The triplets for the relative weightage of quality 

attributes of fruit sample was estimated using the equation (3) 

Q 

A 

RQ 

rel 

= ...(3) 

Q 

sum 

Where, RQ rel 

is the relative weightage of other quality 

attributes of fruits, Q A 

is the triplets for individual attributes 

and Q sum 

is the triplets for sum of the quality attributes. 

Triplets for overall sensory score of fruits 

The overall sensory score of sample was determined 

using the following expression (equation 4). 

SSS 

overall 

= ST RQT 

rel 

+ SC RQC 

rel 

+ SA RQA 

rel 

+ SM RQM 

rel 

...(4) 

j

RAJ & TRIPATHI : Sensory Quality Evaluation of MA Packaged Fruits Applying Fuzzy Logic 197 

Where, SSS overall 

is the overall sensory score of any 

sample, ST, SC, SA and SM are values of triplets for taste, 

color, aroma and mouth feel, RQT rel 

, RQC rel 

, RQA rel 

and RQM rel 

are the triplets for the relative weightage of quality attributes 

namely taste, color, aroma and mouth feel, respectively. Each 

term of the right hand of the equation (4) represents a triplet. 

The following rule was applied (equation 5) for multiplication 

of triplet (a b c) with (d e f). 

(a b c) x (d e f) = (a x d a x e + d x b a x f + d x c) …(5) 

Values of Membership Function of Standard Fuzzy Scale 

The triangular distribution pattern of 6-point sensory 

scale, called standard fuzzy scale is shown in Fig. 2. Symbols 

F1, F2, F3, F4, F5, and F6 represent sensory scales of fruits as: 

not satisfactory / not at all satisfactory, fair / somewhat 

necessary, satisfactory / necessary, good / important, 

very good / highly important and excellent / extremely important 

respectively. Values of membership function for F1 to F6 were 

defined by a set of 10 numbers between 0 - 100 in the 

step of 10. 

1 1 

Value of 

Fuzzy 

membership 

function 

0 

10 

F1 

Not satisfactory / 

Not at all 

necessary 

20 

30 

F2 

Fair / 

Somewhat 

necessary 

40 

Fig. 2. Standard Fuzzy Scale 

50 

F3 

Satisfactory / 

necessary 

F4 

Good / 

important 

F5 

Very good / 

highly 

important 

F6 

Excellent / 

Extremely 

important 

From Fig. 2, the value of membership function for F1 

(Not satisfactory/not at all necessary) was represented as, 

The membership values for MF2, MF3, MF4, MF5 and 

MF6 was obtained 

Values of overall membership function of sensory scores 

on standard fuzzy scale 

The graphical representation of membership function 

of a triplet (a b c) was shown in Fig. 3. It shows that when the 

value of abscissa is ‘a’, value of membership function is 1 and 

when it is less than (a - b) or greater than (a + c), the value is 

zero. For a given value of x on abscissa, value of membership 

function B x 

is expressed by the equation (6) as follows, 

B = 

x 

B = 

x 

 

x- a-b 

 

b 

 

 

a+c -x 

c 

 

 

60 

70 

for a-b < x < a ; 

 

for a < x < a+c 

 

80 

90 

; ...(6) 

For each samples, membership functions (viz. B1, B2, 

B3, and B4) were compared with the membership function of 

standard fuzzy scale (viz., F1, F2, F3, F4, F5, and F6) 

100 

Fig. 3. 

x 

a 

b 

Graphical Representation of Triplet (a b c) and its 

Membership Function 

c 

B x 

1 

Value of membership 

function 

represented a row matrix having 10 elements. Similarity values 

of fruit samples, Sm (F, B) is defined as equation (7) (Das, 

2005; Sinija and Mishra, 2011). 

F o B 

SmF, B = Maximum of F o F and B o B …(7) 

where, F o B is the product of matrix F with transpose of 

matrix B, F o F is the product of matrix F with transpose of F, B 

o B is the product of matrix B with its transpose, and B is the 

values of overall membership function of sensory score for 

fruit samples on standard Fuzzy Scale. 

The MATLAB programme was run to obtain the 

similarity values of different fruit samples. 


Performance of MA packages : 

Apples packed in MA packages shown reduced weight 

loss, retarded the amount of starch formation, preserved good 

colour and firmness in original than fruits stored in normal 

atmosphere (Mangaraj, et. al., 2011b). The modified 

atmosphere of 5% O 2 

and 4% CO 2 

was found to be suitable for 

preservation of guava (cv. Baruipur) (Mangaraj, et al., 2012b). 

MA packaging of EDTA treated litchi fruits enabled the 

reduction of weight loss, prevention of browning, retention 

of good color and sweet taste during extended storage 

(Mangaraj, et. al., 2013). The main constituent of litchi is water 

and its preservation became essential for maintaining fruit 

color and quality. MA packaging reduced the rate of water 

loss from the pericarp by increasing relative humidity around 

the package. Under lower O 2 

and higher CO 2 

(3% O 2 

and CO 2 

) 

storage condition the metabolic and enzymatic activity was 

reduced. 

Fuzzy Logic analysis of sensory data for quality 

evaluation of fruits : 

The sensory data obtained from the judges were 

processed applying Fuzzy logic modeling by developing 

MATLAB programme and important conclusions regarding 

acceptance, ranking, strong and weak attributes of fruit sample 

was drawn. 

 

 

100


Apple (cv. Royal Delicious) : 

The similarity value for all the quality attributes of apple 

is shown in Table 2. It was found that the similarity value for 

sample 1 (PCG-LFR-1) under not satisfactory category (0.0), 

while the same under fair (0.056), satisfactory (0.27698), good 

(0.5408), very good (0.7223) and excellent (0.3516) (Table 2). 

Since similarity value under very good category is the highest 

(0.7223), the overall quality of sample 1 can be considered as 

very good. Using similar reasoning, overall quality of sample 

2 (PCG-LFR-2) is considered as very good having similarity 

value of 0.6918, sample 3 as satisfactory having similarity value 

of 0.72199). Comparing the similarity values of sample 1 and 

sample 2 under very good category it is inferred that sample 1 

is slightly superior to sample 2. Thus the order of ranking of 

MA packed and unpacked apples are sample 1 (very good) > 

sample 2 (very good) > sample 3 (satisfactory). From Table 3 it 

is observed that the similarity value for taste (0.89671) under 

highly important category is the highest. This is followed by 

aroma (important, 0.9545), color (important, 0.9236) and mouth 

feel (necessary, 0.81037). Also the sensory score for aroma is 

higher than that of colour (Table 3). Thus, the order of ranking 

of quality attributes of apple fruit, in general is taste (highly 

important) > aroma (important) > color (important) > mouth 

feel (necessary). 

Table 2. 

Table 3. 

Similarity values of MA packed and unpacked 

Apple fruit sample 

Scale factors Sample 1 

(PCG-LFR-1) 

Sample 2 

(PCG-LFR-2) 

Sample 3 

(CS) 

Not satisfactory, F1 0.0002 0.0005 0.1182 

Fair, F2 0.0559 0.0712 0.4586 

Satisfactory, F3 0.2698 0.2967 0.7199 

Good, F4 0.5408 0.5592 0.5581 

Very good, F5 0.7223 0.6918 0.1912 

Excellent 0.3516 0.3090 0.0116 

Similarity values for quality attributes of apple in 

general 

Scale factors Color Aroma Taste Mouth feel 

Not at all necessary F1 0.0007 0.0013 0.0020 0.0031 

Some what necessary F2 0.0444 0.0641 0.0182 0.1725 

Necessary F3 0.5162 0.6091 0.1345 0.8037 

Important F4 0.9236 0.9545 0.7709 0.6539 

Highly important F5 0.4061 0.3331 0.8671 0.2603 

Extremely important F6 0.0150 0.0214 0.1925 0.0168 

Guava (cv. Baruipur) 

From Table 4 it is found that for sample 4 and 5, the 

similarity value under good category (0.6701 and 0.6646) is 

the highest. Hence the overall quality of these samples is 

good. However, overall quality of sample 6 is fair having 

similarity value of 0.71036. Sample 4 was found to be slightly 

superior to sample 5 in terms of similarity value (Table 4). 

Thus, the order of ranking of guava fruit is sample 4 (good) > 

sample 5 (good) > sample 6 (fair). The comparison of values 

shows that similarity value for taste under highly important 

category (0.8932) is the highest followed by mouth feel 

(important, 0.9026), aroma (important, 0.8874) and color 

(necessary, 0.7924) (Table 5). The sensory score for mouth 

feel was found to be higher than that of aroma. Thus the order 

of preference of quality attributes of guava fruit, in general is 

taste (highly important) > mouth feel (important) > aroma 

(important) > color (necessary). Therefore, these sensory 

attributes are the most desirable characteristics of guava for 

its acceptability and marketing. 

Table 4. 

Similarity values of MA packed and unpacked 

guava sample 


(PCG-LFR-3) 

Sample 5 

(PCG-LFR-4) 

Sample 6 

(CS) 

Not satisfactory, F1 0.0148 0.0211 0.4268 

Fair, F2 0.1720 0.2047 0.7036 

Satisfactory, F3 0.4562 0.4991 0.3801 

Good, F4 0.6702 0.6646 0.2033 

Very good, F5 0.5413 0.4772 0.0633 

Excellent 0.1796 0.1434 0.0026 

Table 5. 

Litchi (cv. Shahi) 

Similarity values of quality attributes of guava fruit 

in general 




Necessary F3 0.7924 0.3691 0.2402 0.3250 

Important F4 0.6183 0.8874 0.7231 0.9026 



The highest similarity values for sample 7, 8, 9 and 10 

are 0.6919, 0.66591, 0.6726 and 0.74389 lie in the overall quality 

category of good, satisfactory, satisfactory and fair, 

respectively (Table 6). On the basis of comparison of the 

similarity values of samples, they are ranked as sample 7 (good) 

> sample 9 (satisfactory) > sample 8 (satisfactory) > sample 10 

(fair). It is observed that (Table 7) the similarity value for color, 

aroma, taste and mouth feel under the category, highly 

important (0.9132), important (0.9705), important (0.91036) and 

necessary (0.81065), respectively is the highest. The similarity 

value for aroma is higher than that of taste both placed in 

important category. Therefore, the order of ranking of quality 

attributes of litchi fruits, in general is colour (highly important) 

> aroma (important) > taste (important) > mouth feel 

Table 6. 

Similarity Values of Litchi fruit sample during 


(PCG-LFR- 

5T) 

Sample 8 

(PCG-LFR- 

5UT) 

Sample 9 Sample 10 

(CS-T) (CS-UT) 

Not satisfactory, F1 0.0197 0.0668 0.0746 0.2414 

Fair, F2 0.2023 0.3380 0.3574 0.7389 

Satisfactory, F3 0.5070 0.6591 0.6726 0.7149 

Good, F4 0.6919 0.6442 0.6365 0.2459 

Very good, F5 0.4826 0.2846 0.2716 0.0129 

Excellent 0.1322 0.0306 0.0276 0.00

RAJ & TRIPATHI : Sensory Quality Evaluation of MA Packaged Fruits Applying Fuzzy Logic 199 

Table 7. 

Similarity values of Quality Attributes of Litchi 

fruits (in general) 




Necessary F3 0.0618 0.3745 0.2436 0.8065 

Important F4 0.6073 0.9705 0.9036 0.7771 



(necessary). It is inferred that colour, taste, and aroma are the 

major quality attributes of litchi. The retention of attractive 

red colour in litchi improves its quality during storage and 

fetches high value in the market (Sivakumar, et al., 2006). 

It was seen that the similarity value for colour under 

very good category (0.65494) is the highest, followed by aroma 

(good, 0.69891), taste (good, 0.6706) and mouth feel 

(satisfactory, 0.71074). Colour is therefore the strongest quality 

of sample 7 followed by aroma and taste. Preserving the 

original colour or improving the same of the litchi fruits can 

enhance its marketability. Thus the quality attributes ranking 

of sample 7 is colour (very good) > aroma (good) > taste 

(good) > mouth feel (satisfactory). 

The sensory data obtained from the judges were 

processed applying Fuzzy logic modeling by developing 

MATLAB programme. The order of ranking of MA packed 

and control stored apple; guava; and litchi were found to be 

sample 1 (very good) > sample 2 (very good) > sample 3 

(satisfactory); sample 1 (good) > sample 2 (good) > sample 3 

(fair); and sample 1 (good) > sample 3 (satisfactory) > sample 

2 (satisfactory) > sample 4 (fair), respectively. The ranking of 

quality attributes of fruits in general were found to be taste 

(highly important) > aroma (important) > color (important) > 

mouth feel (necessary); taste (highly important) > mouth feel 

(important) > aroma (important) > colour (necessary); and color 

(highly important) > aroma (important) > taste (important) > 

mouth feel (necessary) for apple; guava; and litchi, 

respectively. The MA packaged fruits ranked first than that of 

unpacked fruits. 


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Effect of Menopause on Serum Lipid Profile Pattern in Women 

EKTA A.ANDRIYAS, SAPNA SMITH LAL 

1 

Biochemistry 

2 

Deptt. of Clinical biochemistry, Deptt. of Biochemistry 

Higginbottom, Institute of Agriculture : Technology of Science Allahabad, U.P. 

e-mail : sapnaslal@rediffmail.com 

ABSTRACT 

The present study is aimed at comparing the levels of total 

serum cholesterol including their subunits in among the three 

stages of menopause with that of post menopausal women. One 

hundred Fifty apparently healthy, non pregnant female 50 

perimenopausal in Group I, 50 Menopausal in Group II and 50 

postmenopausal Group in III were recruited for the study of 

serum total cholesterol and their sub fractions HDL,LDL, VLDL 

& triglyceride were estimated using enzymatic & established 

mathematical method. There was Significant difference in 

the serum total cholesterol and triglyceride between the three 

groups. There was significant reduction of HDL and VLDL in 

the post menopausal group and a significant increase in the 

level of LDL in the postmenopausal group. The elevated LDL 

and the reduction of cardio protective HDL and VLDL is an 

indication that menopause is an independent risk factor for 

developing CHD in our environment. 

Key word Menopause, CHD, Cholesterol, HDL-Cholesterol, VLDL- 

Cholesterol, LDL-Cholesterol. 

Menopause is a natural event in aging process it signifies 

the end of reproductive phase of life with the cessation of 

cyclic ovarian function manifested by cyclic menstruation 

(Burger, et.al., 2002). This term was originally used to describe 

the reproductive change in human female fertility where the 

end of fertility is traditionally indicated by the permanent 

stoppage of menstruation or mensus. The average age of 

menopause in females is 51 years. Less than 1 % women 

experience it before age of 40 years, with some women under 

going premature menopause at a very early age affecting their 

ability to have children (Derek, 990). Menopause is a normal 

part of life. Changing level of estrogen and progesterone, 

which are the two hormones produced in the ovaries of a 

female may lead to some symptoms which may last for months 

or years. 

After menopause, there is loss of ovarian function. This 

result in adverse change in glucose and insulin metabolism, 

body fat distribution, Coagulation, Fibrinolysis and vascular 

endothelial dysfunction (Spencer. et.al., 1997)There is also 

deranged of lipoprotein profile independent of age 

(Bales,2000).A number of changes that occur in the lipid profile 

after menopause are associated with increased cardiovascular 

disease risk. Lack of estrogen is an essential factor in this 

mechanism. Apart from maintaining friendly lipid profile, 

estrogen changes the vascular tone by increasing nitrous 

oxide production. It stabilizes the endothelial cells, enhances 

antioxidant effects and alters fibrinolytic protein (Taddec, 

et. al., 1996). All these are cardio protective mechanisms, which 

are lost in menopause. 


All experiments was done at laboratory of Faculty of 

Health and Medical Sciences Indigenous System of Medicine, 

Sam Higginbottom Institute of Agriculture Technology and 

Sciences Allahabad. About 5 ml of fasting blood was collected 

from 150 female having age group 40-45 for perimenopause 

stage, 45-50 for menopausal stage, 50 and above for post 

menopausal stage attending the different hospitals of 

Allahabad using sterilized disposable syringe. The blood was 

put into centrifuge tubes; this was allotted to clot and then 

centrifuge at 3000 rpm for 15 min at room temperature. The 

serum obtained was pipetted into clean blood sample and 

analyzed on the day of collection. The serum was analyzed 

for for Serum Total Cholesterol, Serum HDL-Cholesterol, Serum 

LDL-Cholesterol, Serum triglyceride estimation. 


In the present study effect of menopause on lipid profile 

pattern all the three groups were analysed for serum total 

cholesterol, LDL cholesterol and their sub fractions HDL, LDL, 

VLDL and triglyceride. 

The mean, standard deviation and the p value 

significance and t test value for the three stages of menopause 

for Total cholesterol, LDL cholesterol and their sub fractions 

HDL ,LDL, VLDL and triglyceride is shown in Table 1 and the 

p value and t test value for Total cholesterol, LDL cholesterol 

and their sub fractions HDL ,LDL, VLDL and triglyceride in 

comparison to all the three groups is shown in Table 1b, 2b, 

3b, 4b, 5b respectively. During the study it was found that 

there was significant difference in the serum Total cholesterol, 

LDL cholesterol and their sub fractions HDL, LDL, VLDL 

and triglyceride between the three groups. There was a 

significant increase in the Serum Cholesterol, Serum 

Triglyceride, Serum LDL-C, Serum VLDL-C in the three 

menopausal stages. The table shows the increase in the 

menopausal group when compared with the pre menopausal 

group (p value > .000.1 highly significant) and there was also 

increase in the serum total cholesterol, Serum triglyceride,

ANDRIYAS & LAL : Effect of Menopause on Serum Lipid Profile Pattern in Women 201 

Table 1. Effect of menopause on total cholesterol and its sub fractions HDL ,LDL, VLDL and triglyceride 

Groups 

Mean ± S.D. 

Serum Cholesterol Serum Triglyceride Serum HDL-Cholesterol Serum LDL Cholesterol Serum VLDL Cholesterol 

Group I 211.62±18.17 112.7± 16.42 49.58± 18.17 139.5±18.704 42.324± 3.486 

Group II 255.54±17.35 176.6± 24.23 36.16± 17.35 184.06±18.461 51.108± 3.471 

Group III 296.94±25.31 207.64±16.83 22.26± 25.31 233.15±28.328 59.388± 5.072 

Table 2. 

The p value significance and t-test values of serum 

total cholesterol 

Stages P values significance T-Test 

Perimenopausal : Menopausal Statistically significant 12.1756 

(>0.0001) 

Perimenopausal:Postmenopausal Statistically significant 19.1847 

(>0.0001) 

Perimenopausal: Menopausal Statistically significant 

(>0.0001) 

9.5484 

Table 3. 

The p value significance and t-test values of serum 

HDL cholesterol 



(>0.0001) 

Perimenopausal:Postmenopausal Statistically significant 19.2415 

(>0.0001) 


(>0.0001) 

16.1956 

Table 4. 

The pValue and t Test value of Serum Triglyceride 

Stages 

Perimenopausal : Menopausal 

Perimenopausal:Postmenopausal 

Perimenopausal: Menopausal 

Table 5. 

Table 6: 

P values significance T-Test 

Statistically significant 15.3724 

(>0.0001) 


(>0.0001) 


(>0.0001) 

The pValue and t Test value of Serum LDL 

Cholesterol 

Stages P value significance T Test 

Perimenopausal : Menopausal Statistically 12.0143 

significant (>0.0001) 

Perimenopausal:Postmenopausal Statistically 19.6438 


Perimenopausal: Menopausal Statistically 


10.3835 

Showing the pValue and t Test value of Serum 

VLDL Cholesterol 



(>0.0001) 

Perimenopausal :Postmenopausal Statistically significant 19.6096 

(>0.0001) 


(>0.0001) 

9.5309 

Serum LDL-C, Serum VLDL-C concentrations in the 

postmenopausal women when compared with menopausal 

women (p value > .000.1 highly significant). 

But on the other hand the study shows significant 

decrease in the Serum HDL-Cholesterol in the three menopausal 

stages. The Table 1 shows the decrease in the menopausal 

group when compared with the pre menopausal group 

(p value > .000.1 highly significant) and there was also decrease 

serum HDL-Cholesterol in the concentrations in the 

postmenopausal women when compared with menopausal 

women. (p value > .000.1 highly significant). 

Lipid profiles are affected by metabolic conditions and 

alteration in lipid metabolism have been implicated in 

atherosclerosis and coronary heart disease .results from this 

study on lipid profile indicate that menopause alter the lipid 

profile in women .the total cholesterol, LDL-C Triglyceride 

and VLDL-C were significantly higher and HDL-C lower in the 

post menopausal women a similar observation. Barett and 

Bush 1993 in post, menopausal Caucasians. The elevated 

TC,LDL-C in post menopausal women and women greater 

than 45 yrs has been attributed to hormonal changes and 

failure of follicular development ,where the plasma estradoil 

levels that reduces the risk of coronary heart disease 

Mean values 

300 

250 

200 

150 

100 

50 

0 

211.62 

255.54 

296.94 

Pre-M Meno Post-m 

Stages of menopause 

Fig. 1. Effect of menopause on Serum Total Cholesterol 

Mean values 

50 

45 

40 

35 

30 

25 

20 

15 

10 

5 

0 

49.58 

36.16 

22.26 

Pre-M Meno Post-m 

Stages of menopause 

Fig. 2. Effect of menopause on Serum HDL-Cholesterol


falls below the levels seen in the premenopausal women. 

Unfavorable Changes in HDL-C,LDL-C after menopause have 

been independently reported by (Pascot et. al., 1999: Gardy 

et. al., ,1992).The TC and LDL-C were significantly higher 

and HDL-C Lower in women above 45 yrs when compared to 

those of women aged between 25-45 yrs. Increasing age has 

been associated with higher plasma LDL-C ,where significantly 

higher LDL-C was observed in postmenopausal women than 

in premenopausal women (Schaefer, et. al., 1994) also, 

observed high total cholesterol, LDL-C and VLDL-C as well 

as triglyceride levels with increasing age. 

In the present study there was significant differences in 

the total cholesterol going to the higher side with the increase 

in age but there was significant reduction in the in the cardio 

protective HDL-C and significant increase in the 

atherosclerotic LDL-C and VLDL-C. This is in agreement with 

findings in other studies (Jenson, et. al., 1990, Edr and Gidez, 

1992, Igweh and Aloamaa, et. al., 2003). It has been also 

estimated that for any 1mg/dl(0.026mmol/ml) increase in HDL- 

C there is a 3% decrease in the risk of coronary artery disease 

and a 4.7% decrease of mortality from cardio vascular disease 

(Okonofua, 1990.) 

Several studies have shown the beneficial effect of 

hormonal replacement therapy on the lipid profile of 

menopausal women (Abbot, et. al., 1988. Stampfer, et al., 

1991). 

Observational studies over years have touted the 

beneficial effects of hormone replacement therapy (HRT)in 

preventing coronary artery disease in the post menopausal 

women (Hulley et. al., 1998). More recent studies have 

however ,cast some doubts on the beneficial effect of HRT 

especially in patient with established cardiovascular disease 

(Cheng, 2000). Further studies are needed in this area. 

It should be noted that post menopausal women have 

unfriendly lipid profile, it is thus to counsel or proper dietary, 

social and physical habits. 


I am highly grateful to Prof. Dr. R. B. Lal, Vice Chancellor 

of Sam Higginbottom Institute of Agriculture Technology and 

Sciences Allahabad, for providing me necessary facilities for 

research work. I am also grateful to staff of Hays Memorial 

Mission Hospital for providing me samples and other 

information. 


Abbot, R.D., Wilson, P.W.F., Kannel, W.B. and B. Castelli, W.P. 1988. 

High- density lipoprotein cholesterol, total cholesterolscreening 

and myocardial infarction. The Framingham study. Arterosclerosis; 

8: 207. 

Bales, A.C. 2000. In search of lipid balance in older women; New studies 

raise questions about what works best.Postgrad. Med.; 108(7): 57-72. 

Barrett Connor, E.,T. L. Bush 1993. Estrogen therapy and Coranory 

heart disease in women. JAMA, 265: 1861-1867. 

Burger E.F., Kustin A.A. and Baron, D.C. 2002. Endocinology and 

metabolism. Ann. Int. Med. 251: 872-942. 

Cheng, G.S. 2000. Cardiac events increased in the first 2 years of HRT. 

Intern. Med. News; 33(9): 1-2. 

Derek L.J., 1990. Menopause. Fundamentals of Obstetrics and 

gynaecology, Springfield Illinois, U.S.A. 2nd edition, pp. 71-72. 

Edr, H.A., and Gidez, L.L. 1982. The Clinical significance of the plasma 

high density lipoprotein. Med. Clin. North Am., 66: 431-434. 

Gardy. D.S.M. Robin, and Petitti D.B. 1992. Expert panel on detection 

and evaluation and treatment of high blood cholesterol in adults. 

Arch.int.med. 148: 36-69. 

Hulley, S., Grady, D., Bush, T. 1998. The randomized trial of estrogen 

plus progestin for secondary prevention of coronary heart disease 

in postmenopausal women: Heart and Estrogen/progestin 

replacement study (HERS) Research group. JAMA 7: 605-13. 

Igweh, J.C., Aloamaka, C.P., 2003. HDL/LDL Ratio- A Significant, 

Predisposition to the Onset of Atherosclerosis. njhbs, 2:(2) 78-82. 

Jensen, J., Nilas, L., Christiansen, C. 1990. Influence of menopause on 

serum lipid and lipoprotein. Matruita, 12: 321-31. 

Okonofua, E.E., Lawal, A., Bamgbose, J.K. 1990. Features of 

Menopause and Menopausal age in Nigerian women. Int. J. Gynaecol. 

Obstet., 31(4): 341-5. 

Pascot. A., Lemieux. S., Lemieux. I., Prudhomme, Tremblay. A., 

Bouchard. C., Nadeau. A., Oulliard. C., Tchemof. A., Bergeron. J. 

and Despres. J. P. 1999. Influence of perimenopause on 

cardiovascular risk factors and symptoms of middle age healthy 

women. Arch.int.Med., 154: 2349-2355. 

Schaefer, E. J., Lamon-Fava, S., Johnson, S., Ordovas, J.M., Schaefer, 

M.M., Castelli, W.P. and Wilson, P.W. 1994. Effects of gender and 

menopausal status on the association of apolipoprotein Ephenotype 

with plasma lipoprotein levels J. American Heart Association, 

14: 1105-1113 

Spencer, C.P., Godsland, H, Stevenson 1997. There a menopausal 

metabolicsyndrome Gynecol.Endocrinol.; 11: 341-355. 

Stampfer, M.J., Colditz, G.A., Willet, W.C. 1991. Postmenopausal 

estrogen therapy and cardiovascular disease. Ten year follow up 

from Nurses Health Study.N. Engl. J.Med., 325(11): 956-62 

Taddec, S., Virdis, A., Ghiadoni, L., Mattec, P., Sudano, I., Bernini, 

G. 1996. Menopause is associated with endothelial dysfunction in 

women. Hypertension. 28: 576-582. 



Sensory Evaluation of Vegetables Grown Under Organic and Inorganic Conditions 

1 

BAJPAI PREETI AND 2 PUNIA DARSHAN 

1 

Dept. of Foods and Nutrition, College of Home Science, MPUAT, Udaipur, Rajasthan 313 001 

2 

Department of Foods and Nutrition, I. C. College of Home Science, Chowdhry Charan Singh (CCS) Haryana 

Agricultural University, Hisar Haryana 125 004 

e-mail: preeti.preetibajpai@gmail.com 

ABSTRACT 

The present investigation was carried out with the objectives 

to study sensory characteristics of vegetables grown under 

organic and inorganic conditions. The recipes i.e. carrot 

vegetable, fenugreek leaves vegetable, garlic pickle, okra 

vegetable, Singare vegetable, tomato chutney and tomato puree 

prepared by using organically grown , inorganically grown 

and conventionally grown vegetables were subjected to sensory 

evaluation by a panel of ten judges using 9-point hedonic scale 

The results of the present study revealed significant differences 

for the sensory characteristics between organically and 

inorganically grown vegetables Sensory evaluation of vegetable 

revealed that recipes prepared using organically, grown 

vegetables scored better scores for some of the characteristics 

as compared to their inorganically and conventionally grown 

counterparts 

Key words Vegetables, Nutritional evaluation, Organic, Inorganic 

Organic agriculture is a unique production management 

system which promotes and enhances agro-ecosystem health, 

including biodiversity, biological cycles and soil biological 

activity. Organic agriculture excludes the use of synthetic 

pesticides, conventional fertilizers, pharmaceuticals and by 

definition excludes genetically modified plants and animals 

(Roitner-Schobesberger, et. al., 2008). 

Public interest is increasingly focusing on the problem 

of the quality of foods because of people’s growing awareness 

of health and the environment. Organic food products with 

high nutritive value and without chemicals (with potential 

carcinogenic and mutagenic properties) are being increasingly 

preferred over conventional agro products, which are 

cultivated using insecticides, pesticides and chemical 

fertilizers. 

Survey indicates that consumers purchase organic 

produce because of the belief that they are more nutritious 

than conventionally grown foods (Winter and Davis, 2006). 

Fresh organic foods are even more nutritious because of higher 

percentage of vitamins and minerals. Most importantly, fresh 

organic foods are free from any kind of chemical residues .On 

an average, organic food contain higher level of Vitamin C 

and essential minerals such as calcium, magnesium, iron and 

chromium. Organic spinach, lettuce, cabbage and potatoes 

contain particularly high levels of minerals (Worthington 2001). 

Numerous studies confirm that many people believe that 

organic foods are healthier and safer than inorganically 

produced foods and are produced in a more environmentally 

compatible manner (Rembialkowska, 2007; Roitner- 

Schobesberger, et. al., 2008). Although the interest in the 

organically grown foods has been on increase, there have not 

been substantial studies to substantiate convincingly that 

organically grown foods are nutritionally superior to their 

inorganically grown counterparts. 

Owing to the nutritional importance of vegetables in 

our diet and increasing concern toward health and organic 

farming of people’s it becomes important to evaluate the 

nutrient composition of vegetables grown under organic and 

inorganic conditions. 

The present investigation tried to determine the sensory 

characteristics of vegetables grown under organic and 

inorganic conditions. 


Procurement of vegetables : 

The samples of the vegetables viz. tomato, okra, garlic 

and fenugreek leaves grown under organic and inorganic 

conditions were procured from Vegetables Farm, Chaudhry 

Charan Singh Haryana Agricultural University, Hisar. The 

vegetable viz. carrot and singare (Siliqua of radish) were 

procured from Domestic Farm, College of Home Science, 

Chaudhry Charan Singh Haryana Agricultural University, 

Hisar. The vegetable above mentioned vegetables were also 

procured from the local market for comparative study. 

Product development and sensory evaluation : 

Development of products : 

The recipes i.e. Singare veg., carrot veg., garlic pickle., 

fenugreek leaves veg., okra veg., tomato chutney, and tomato 

puree were prepared from the vegetables viz. Singare, carrot, 

garlic, fenugreek leaves, okra and tomato, grown organically, 

inorganically and conventionally. 

Sensory evaluation : 

The recipes i.e. Singare vegetable, carrot vegetable, 

fenugreek leaves vegetable, garlic pickle, okra vegetable, 

tomato chutney and tomato puree prepared by using


organically grown , inorganically grown and conventionally 

grown vegetables were subjected to sensory evaluation by a 

panel of ten judges using 9-point hedonic scale where: 1 = 

dislike extremely, 2 = Dislike very much, 3 = Dislike moderately, 

4 = Dislike slightly, 5 = neither like nor dislike, 6 = Like slightly, 

7 = Like moderately, 8 = Like very much and 9 = like extremely. 

Panelists were asked to comment on liking of color, texture, 

flavor and overall acceptability in morning time. 


The recipes viz., Singare vegetable, carrot vegetable, 

fenugreek leaves vegetable, garlic pickle, okra vegetable, 

tomato chutney and tomato puree were prepared using 

organically grown , inorganically grown and conventionally 

grown vegetables and were organoleptically evaluated. The 

results obtained on the sensory characteristics of various 

products are discussed below. 

Carrot vegetable prepared from organically grown carrot 

was liked very much in terms of all sensory parameters, 

including color, appearance, aroma, texture, taste and overall 

acceptability. However vegetable prepared from inorganically 

and conventionally grown carrot were ‘liked moderately’ in 

terms of all sensory attributes. Carrot vegetable prepared from 

organically grown carrot was found superior in terms of its 

color, appearance, aroma, texture, taste and overall 

acceptability as compared to carrot vegetable, prepared from 

inorganically and conventional grown carrot. 

Mean scores of sensory characteristics revealed that 

pickle made from organically grown, inorganically grown 

conventionally grown garlic was “liked moderately” in terms 

of color, appearance, aroma, texture, taste and overall 

acceptability by the judges. 

Mean scores of sensory characteristics indicated that 

vegetable prepared using organically, inorganically and 

conventionally grown Singare was “liked moderately” in terms 

Table 1. Sensory characteristics of vegetable recipes 

Carrot vegetable 

Garlic pickle 

Singare vegetable 

Fenugreek leaves 

vegetable 

Okra vegetable 

Tomato chutney 

Tomato puree 

Conditions Color Appearance Aroma Texture Taste Overall 

Acceptability 

Organic 8.40 ± 0.22 8.20 ± 0.25 8.30 ± 0.16 81.00 ± 0.28 8.20 ± 0.25 8.24 ± 0.09 

Inorganic 7.60 ± 0.16 7.50 ± 0.17 7.40 ± 0.16 74.00 ± 0.16 7.50 ± 0.17 7.48 ± 0.04 

Conventional 7.60 ± 0.16 7.60 ± 0.22 7.60 ± 0.16 74.00 ± 0.28 7.30 ± 0.15 7.50 ± 0.07 

CD(P?0.05) 0.53 0.62 0.58 0.7 0.56 0.2 

Organic 8.00 ± 0.33 7.90 ± 0.31 7.80 ± 0.36 7.80 ± 0.35 8.0 ± 0.33 7.90 ± 0.24 

Inorganic 7.30 ±0.33 7.10 ± 0.31 7.30 ± 0.30 7.50 ± 0.27 7.30 ± 0.30 7.30 ± 0.24 

Conventional 7.50 ±0.22 7.40 ± 0.22 7.30 ± 0.30 7.10 ± 0.28 7.20 ± 0.36 7.30 ± 0.23 

CD NS NS NS NS NS NS 

Organic 7.90 ± 0.10 7.90 ± 0.10 7.60 ± 0.22 7.50 ± 0.17 7.50 ± 0.17 7.70 ± 0.10 

Inorganic 7.30 ± 0.33 7.80 ± 0.13 7.80 ± 0.13 7.50 ± 0.17 7.80 ± 0.20 7.74 ± 0.09 

Conventional 7.50 ± 0.22 7.90 ± 0.10 7.90 ± 0.18 7.90 ± 0.10 7.40 ± 0.16 7.78 ± 0.09 

CD(P?0.05) 0.36 NS NS NS NS NS 

Organic 7.80 ± 0.20 8.00 ± 0.15 7.60 ± 0.27 7.60 ± 0.22 7.70 ± 0.21 7.68 ± 0.18 

Inorganic 7.50 ± 0.17 7.40 ± 0.16 7.40 ± 0.16 7.30 ± 0.15 7.40 ± 0.16 7.40 ± 0.15 

Conventional 7.50 ± 0.17 7.70 ± 0.14 7.40 ± 0.16 7.30 ± 0.15 7.30 ± 0.15 7.50 ± 0.11 

CD(P?0.05) NS 0.45 NS NS NS NS 

Organic 7.90 ± 0.23 8.10 ± 0.18 7.90 ± 0.23 8.00 ± 0.21 8.20 ± 0.20 8.02 ± 0.19 

Inorganic 7.60 ± 0.16 7.60 ± 0.22 7.60 ± 0.16 7.50 ± 0.17 7.80 ± 0.13 7.62 ± 0.14 

Conventional 7.50 ± 0.17 7.50 ± 0.17 7.40 ± 0.16 7.20 ± 0.20 7.50 ± 0.17 7.42 ± 0.12 

CD(P?0.05) NS 0.55 NS 0.56 0.5 0.45 

Organic 7.80 ± 0.20 7.70 ± 0.15 7.70 ± 0.15 7.80 ± 0.20 8.00 ± 0.21 7.80 ± 0.17 

Inorganic 7.40 ± 0.22 7.30 ± 0.15 7.20 ± 0.13 7.20 ± 0.14 7.40 ± 0.22 7.30 ± 0.14 

Conventional 7.80 ± 0.20 7.80 ± 0.20 7.60 ± 0.16 7.70 ± 0.21 7.74 ± 0.20 7.74 ± 0.13 

CD(P?0.05) NS NS 0.44 0.54 NS 0.44 

Organic 8.20 ± 0.21 8.10 ± 0.27 7.80 ± 0.20 7.50 ± 0.17 7.50 ± 0.17 7.98 ± 0.16 

Inorganic 7.30 ± 0.13 7.40 ± 0.18 7.80 ± 0.20 7.90 ± 0.18 7.90 ± 0.05 7.50 ± 0.05 

Conventional 7.80 ± 0.13 7.70 ± 0.21 7.80 ± 0.20 7.50 ± 0.17 7.60 ± 0.16 7.68 ± 0.10 

CD(P?0.05) 0.47 0.64 NS NS NS 0.34 

Values are mean ± SE of ten independent observations. 

NS = Not significant

PREETI & DARSHAN : Sensory Evaluation of Vegetables Grown Under Organic and Inorganic Conditions 205 

appearance, aroma, texture, taste and overall acceptability 

differed significantly except the scores for the color of 

vegetable made from organically grown Singare significantly 

higher as compared to vegetable made from inorganically 

grown Singare. Though the color of Singare vegetable 

prepared from organically grown Singare scored significantly 

higher score than that of inorganically and conventionally 

grown singare yet the color of the vegetables liked moderately 

The mean score presented in Table 1 showed that 

appearance of the vegetable prepared from fenugreek leaves 

obtained organically grown leaves ‘liked very much’. The 

scores for all other sensory attributes of the vegetables were 

almost similar and all the three types of vegetables were ‘liked 

moderately’. 

The okra vegetable prepared from organically grown 

okra was ‘liked very much’ in terms of appearance, texture, 

taste and overall acceptability but the color and aroma of the 

vegetable were “liked moderately” by the judges. Okra 

vegetable prepared from inorganically and conventionally 

grown okra were liked moderately in terms of all sensory 

characteristics. 

Chutney prepared from organically grown tomatoes was 

‘liked moderately’ by the panel member in terms of color and 

appearance, aroma texture and overall acceptability but it was 

‘liked very much’ in terms of taste; However, chutney prepared 

from inorganically grown and conventionally grown tomatoes 

was ‘liked moderately’ in terms of all sensory characteristics 

by a panel of ten judges. Tomato chutney made from organically 

grown tomatoes was found superior in terms of aroma, texture, 

and overall acceptability as compared to the chutney prepared 

using inorganically grown tomatoes. 

On the basis of mean scores of sensory characteristics 

it was found that puree prepared from inorganic tomatoes 

was “liked moderately” in terms of all the sensory attributes, 

whereas puree prepared from organic tomatoes was found to 

be “liked very much” in terms of color and appearance but it 

was “liked moderately” in terms of aroma, taste, texture and 

overall acceptability. Tomato puree prepared from 

conventional tomatoes was “liked moderately” by the judges 

in terms of all sensory attributes including color, appearance, 

aroma, texture and taste. 

Rembialkowska, 2007 found better scores for the taste 

and other organoleptic attributes of the organically grown 

carrots. Better total flavor strength, sweet taste in organically 

grown carrot was reported by Hogstad, et al., 1997. Soups 

made from organic fertilized okra were judged more acceptable 

(Taiwo, et al., 2002). Organically grown tomatoes were 

perceived by the panelist to be superior for their taste, texture 

and flavor as compared to inorganically grown (McCollum et 

al., 2005 and Heeb, et.al., 2004). Shankaro and Sumathi, 2008 

reported similar findings for tomato puree made from 

organically grown and inorganically grown tomatoes. 

Better sensory scores for the color of Singare vegetable 

made from organically grown, Singare (7.90) were observed 

than theirs inorganically grown (7.30) counterparts. Mean 

scores for the appearance of the organically grown fenugreek 

leaves vegetable was significantly higher (8.00) as compared 

to mean scores of their inorganic counterparts (7.40). 

Organically grown okra vegetable had significantly higher 

sensory scores for its appearance (8.10), texture (8.00), taste 

(8.20) and overall acceptability (8.02) as compared to its 

conventional (7.5, 7.4, 7.2, 7.5, and 7.42, respectively) 

counterpart. Tomato chutney made from organically grown 

tomatoes found significantly superior in terms of aroma (7.70), 

texture (7.8) and overall acceptability (7.8) as compared to the 

chutney prepared using inorganically grown tomatoes (7.2, 

7.2, and 7.30, respectively). On the basis of mean scores of 

sensory characteristics it was found that puree prepared using 

organic tomatoes had significantly better sensory scores for 

color, appearance and overall acceptability as compared to 

the scores of their inorganic counterparts. 


Heeb, A., Lundegardh, B., Ericsson, T. and Savage, G. P. 2004. Nitrogen 

form affects yield and taste of tomatoes. Journal of the Science of 

Food and Agriculture, 85(8): 1405-1414. 

Hogstad, S., Risvik, E., and Steinsholt, K. 1997. Sensory quality and 

chemical composition in carrots: a multivariate study. Acta 

Agriculturae Scandinavica, 47: 53-264. 

McCollum, T. G., Chellemi, D. O., Rosskopf, E. N., Church, G. T. 

and Plotto, A. 2005. Postharvest quality of tomatoes produced in 

organic and conventional production systems. Hort. Science, 40(4): 

959-963. 

Rembialkowska, E. 2007. Quality of plant products from organic 

agriculture. J. Sci .Food Agri., 87: 2757-2762. 

Roitner-Schobesberger, B., Darnhofer, I., Somsook, S. and Vogl, C. 

2008. Consumer perceptions of organic foods in Bangkok, Thailand. 

Food Policy., 33:112–121. 

Shankaro, K. and Sumathi, S. 2008. Effect of organic farming on 

nutritional profile of selected vegetable crops. Karnataka J. Agric. 

Sci., 20 (3):586-588. 

Taiwo, L. B., Adediran, J. A., Ashaye, O. A., Odofin, O. F., and Oyadoyin, 

A. J. 2002. Organic okro (Abelmoschus esculentus): its growth, 

yield and organoleptic properties. Nutrition & Food Science, 32(5): 

180-183. 

Winter, C. K. and Devis, S. F. 2006. Organic foods. J. Sci. Food Agri. 

71: 117-124 

Worthington, V. 2001. Nutritional quality of organic versus conventional 

fruits, vegetables and grains. J. Alt. Compl. Med., 7(2):161–173. 



Hypolipidemic Potential of Bacopa monniera in Cholesterol Fed Rats 

SYED MANSOOR ALI AND GYAN CHAND JAIN 1 

Department of Zoology, University of Rajasthan, Jaipur 302 004, India 

e-mail: jain_gc@yahoo.co.in 

ABSTRACT 

In the present study the effect of 70% ethanolic extract of Bacopa 

monniera Linn. (Scrophulariaceae) whole plant was evaluated 

on serum and hepatic lipid profiles of hyperlipidemic rats. 

Experimental hyperlipidemia was induced by feeding 

cholesterol (500 mg/kg b.wt./day) suspended in coconut oil for 

60 days. Feeding with cholesterol resulted in a significant 

(P

ALI & JAIN et. al., : Hypolipidemic Potential of Bacopa monniera in Cholesterol Fed Rats 207 

Autopsy : 

At the end of the experiment rats were weighed and 

fasted overnight. These were sacrificed under mild ether 

anesthesia. Blood sample was collected by cardiac puncture. 

The blood was allowed to clot at room temperature and the 

serum was separated by centrifugation and stored at - 20 0 C 

until assayed. 

Liver was quickly excised, cleaned, washed by chilled 

normal saline, and immediately frozen at -70 0 C for biochemical 

analysis. 

cholesterol ratio was observed as compared with cholesterol 

fed control rats. 

Serum biochemistry : 

Serum samples were analyzed for total cholesterol (TC) 

(Zlatkis, et. al., 1953), Low density lipoprotein cholesterol 

(LDL-cholesterol) (Friedwald, et. al., 1972), high density 

lipoprotein cholesterol (HDL-cholesterol) (Burnstein, et. al., 

1970) triglycerides (TG) (Gottfried and Rosenberg, 1973) and 

phospholipid (Zilversmit and Davis, 1950). Atherogenic index 

was calculated from the following formula: 

Atherogenic index = HDL - cholesterol : total cholesterol. 

Tissue biochemistry : 

Liver total cholesterol, triglycerides and phospholipid 

were measured by the methods as described above for serum. 

Fecal cholesterol : 

Fecal matter of control and treated rats was collected 

daily during the last week of the experiment and dried at 40 0 C. 

The dried fecal matter was used for the analysis of total 

cholesterol (Zlatkis, et. al., 1953). 

The results are expressed as mean + SEM Student “t” 

test was used to determine the significance of differences 

between various groups. Values of P


Fecal cholesterol : 

There was significant (P

ALI & JAIN et. al., : Hypolipidemic Potential of Bacopa monniera in Cholesterol Fed Rats 209 

to better understand the mechanisms of action of this medicinal 

plant. 


The authors are grateful to Prof. N.P. Singh, Head of the 

Department and Prof. N. K. Lohiya, Coordinator, SAP-UGC, 

Department of Zoology, University of Rajasthan, Jaipur for 

providing necessary facilities and for the award of SAP-JRF 

to S. M. Ali. 


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Chemoattraction in Entomopathogenic Nematodes 

RASHID PERVEZ 1 AND S. S. ALI 2 

2 

Indian Institute of Pulses Research, Kanpur (U.P.) India 208 024 

1 

Indian Institute of Spices Research, Kozhikode (Kerala) - 673 012 

e-mail: rashid_pervez@rediffmail.com 1 , ss_ali@rediffmail.com 2 

ABSTRACT 

Attraction of infective juveniles of entomopathogenic 

nematodes, Steinernema seemae and S. carpocapsae towards 

lepidopteran insect pests viz., 2 nd instar larva of gram pod borer, 

Helicoverpa armigera, legume pod borer, Maruca vitrata and blue 

butterfly, Lampides boeticus were tested in petridish. Results 

show that, both species of EPNs were found positive attraction 

responses towards test insects. Among test EPNs, S. seemae was 

found more responsive towards insect pests than S. carpocapsae. 

The maximum attraction of S. seemae was recorded towards H. 

armigera (81 %), followed by L. boeticus (62 %) and M. vitrata 

(41%). In case of S. carpocapsae, maximum attraction responses 

towards H. armigera (60 %), whereas least towards M. vitrata 

(41 %). The attraction behaviour of entomopathogenic 

nematodes vary between species to species. 

Key words Entomopathogenic nematodes, Attraction, Behaviour. 

Host recognition by EPNs is due to various biotic and 

abiotic stimuli. Chemicals cues and behavioural responses to 

these cues play a major role in inter specific interactions and 

host finding activities of EPNs. Bilgrami 2004 reported that, 

attraction, aggregation and preferential behaviour of EPNs 

involving sensory responses are vital steps in their life cycle. 

EPNs utilize and recognize signals from both categories 

to find their host, depending upon their action on receiver 

insect, allelochemicals may be categorised into allomones, 

synomones and kairomones (Bilgrami, 2004). Role of allomones 

and kairomones has been well defined for host searching, 

attraction and aggregation behaviour of nematodes is still 

mistily known. 

In view of the sparse knowledge on the attraction 

behaviour of EPNs, present studies on the attraction and 

preferential behaviour of S. seemae (Ali, et al., 2005a) and S. 

carpocapsae (Weiser, 1955) Wouts et al., 1982 towards 2 nd 

instar larva of gram pod borer, Helicoverpa armigera, legume 

pod borer, Maruca vitrata and blue butter fly Lampides 

boeticus. 


Nematode and insect cultures: S. seemae and 

S. carpocapsae were taken from Nematology laboratory of 

this institute and test insects, H. armigera, M. vitrata and 

L. boeticus larvae were collected from standing crop from the 

IIPR Experimental Farm and CSAU & T experimental field. 

The larvae were sorted out and those of same size were taken 

for present study. 

Bioassay: The attraction of S. seemae and 

S. carpocapsae towards 2 nd instar larva of H. armigera, 

M. vitrata and L. boeticus was worked out through petridish 

method. A Petridish 5.5 cm in diameter was divided into three 

zones by drawing two concentric circles of 0.5 cm and 2.5 cm 

in diameter. The later circle was termed as reference circle. A 

plastic straw pipe with a small piece of filter paper glued at 

one end was placed vertically in the inner circle of petri-dishes. 

Water agar (1.5 %) is poured into the petridishes and straw 

pipe to make a required thickness (4 mm) of agar. One larva of 

test insect was placed in straw pipe and seal with cellotape. 

Five infective juveniles of test species of EPN was released at 

various places at the periphery of the middle circle (reference 

circle) and their distribution was recorded after 2 h. All 

experiment was done at room temperature along with control 

(without insect) and replicated ten times. 

The area of each zone and weighting factors was 

calculated (Table 1) by dividing the area of outer zone with 

each of the three zones and score were obtained by summing 

up the product of the number of worms in each zone with their 

corresponding weighting factors. These scores were then 

converted into percentage (Table 2). 


Results show that, both species of EPNs were found 

positive attraction responses towards H. armigera, M. vitrata 

and L. boeticus. Among test EPNs, S. seemae was found more 

responsive towards insect pests compared to S. carpocapsae. 

The maximum attraction of S. seemae was found towards H. 

armigera (81 %), followed by L. boeticus (62 %) and M. vitrata 

(42%). In case of S. carpocapsae, maximum attraction 

responses was recorded towards H. armigera (62 %), whereas 

least attraction responses towards L. boeticus (42 %) and M. 

vitrata (41 %) (Fig. 1). 

The attraction behaviour of entomopathogenic 

nematodes vary between species to species and individual to 

individual. They use strategies to find hosts on the basis of 

ambusher and cruiser behaviour (Campbell, et al., 2002). 

Among ambusher, some EPNs species nictate, or raise their 

bodies off the soil surface so they are better poised to attach

Table 1. 

PERVEZ & ALI : Chemoattraction in entomopathogenic nematodes 211 

Area and weighting factors for the zones marked 

on the agar plates 

Inner zone Middle zone Outer zone 

Area (Sq cm) 0.20 4.90 23.74 

Weighting factors 118.70 4.84 1.0 

Table 2. 

Twenty one positions nematodes can occupy with 

their ranks and score 

Position Inner 

zone 

Middle 

zone 

Outer 

zone 

Ranks 

zone 

Scores Attraction 

responses (%) 

1. 5 0 0 21 593 100 

2. 4 1 0 20 479 81 

3. 4 0 1 19 475 80 

4. 3 2 0 18 366 62 

5. 3 1 1 17 362 61 

6. 3 0 2 16 358 60 

7. 2 3 0 15 252 42 

8. 2 2 1 14 247 42 

9. 2 1 2 13 244 41 

10. 2 0 3 12 240 41 

11. 1 4 0 11 138 23 

12. 1 3 1 10 134 23 

13. 1 2 2 9 130 22 

14. 1 1 3 8 126 21 

15. 1 0 4 7 123 21 

16. 0 5 0 6 24 4 

17. 0 4 1 5 20 4 

18. 0 3 2 4 16 3 

19. 0 2 3 3 13 2 

20. 0 1 4 2 9 2 

21. 0 0 5 1 5 1 

to passing insects (Campbell and Gaugler, 1997), while, many 

EPNs are able to jump by forming a loop with their bodies that 

creates stored energy (Campbell and Kaya, 2000). Other 

species adopt a cruising strategy and rarely nictate. Instead, 

they roam through the soil searching for potential hosts. These 

foraging strategies influence which hosts the nematodes 

infect. 


The authors expresses their gratitude’s to Director, 

Indian Institute of Pulses Research, Kanpur, for providing all 

the facilities for this study. This study was supported by a 

grant from the Department of Science and Technology (DST), 

Fig. 1. 

Attraction of entomopathogenic nematodes towards 

lepidopteran insect pests 

Ministry of Science & Technology, Government of India, New 

Delhi, which is gratefully acknowledged. 


Ali, S. S., Shaheen, A., Pervez, R. and Hussain, M. A. 2005a. Steinernema 

masoodi sp. n. and Steinernema seemae sp. n. (Rhabditida: 

Steinernematidae) from Uttar Pradesh, India. International Journal 

of Nematology, 15(1) : 89 – 99. 

Bilgrami, A. L. 2004. Host searching and attraction behaviour of 

entomopathogenic nematodes. International Journal of 

Nematology, 14(1): 23- 29. 

Bilgrami, A. L. and Pervez, R. 2000. Prey searching and attraction 

behavior of Mesodorylaimus bastiani and Aquatides thornei 

(Nematoda: Dorylaimida). International Journal of Nematology, 

10 (2): 199- 206. 

Campbell, J. F. and Gaugler, R. R. 1997. Inter-specific variation in 

entomopathogenic nematode foraging strategy: dichotomy or 

variation along a continuum. Fundamental and Applied 

Nematology. 20(4): 393-398. 

Campbell, J. F. and Kaya, H. K. 2000. Influence of insect associated 

cues on the jumping behavior of entomopathogenic nematodes 

(Steinernema spp,). Behaviour, 137(5): 591-609. 

Received on 03.08.2012 Accepted on 05.09.2012


Effectiveness and Economics of Integrated Weed Management in Transplanted Rice 

(Oryza sativa) 

BIRENDRA KUMAR 1 , RANVIR KUMAR 2 , SUMAN KALYANI 3 AND M. HAQUE 4 

1,4 

Department of Agronomy, Bihar Agricultural College, Sabour, Bhagalpur, Bihar 813 210 

2 

Deptt. of Agril. Economics, B.P.S. Agricultural College, Purnea City, Purnea, Bihar 854 302 

3 

Deptt. of Plant Breeding & Genetics, B.P.S. Agril. College, Purnea City, Purnea, Bihar 854 302 

e-mail : ranvir.bausabour@gmail.com 

ABSTRACT 

In experimental field, Cyperus rotundus (L)., Cyperus iria (L), 

Cyperus difformis (L)., Fimbristylis miliacea (L) Vaha., 

Echinochloa colona ( L).Link, Echinochloa crus-galli (L) 

P. Beauv, and Commelina benghalensis (L) were the dominant 

weed flora species in rice fields. Manual weeding at 20 and 40 

days after transplanting (DAT), butachlor @1.5 kg a.i/ha preem 

followed by (fb) 2,4-D easter 0.5 kg a.i./ha Post Emergence 

(POE), butachlor @ 1.5 kg a.i/ha as pre-em fb azimsulfuron @ 

25.0 g a.i/ha POE, bispyribac 12.5 g a.i/ha + azimsulfuron 12.5 

g a.i/ha POE, butachlor @ 1.5 kg a.i/ha as pre-em fb 

ethoxysulfuron @ 15 g a.i/ha as POE proved equally effective 

in increasing most of the growth parameters, yield attributes, 

yield and economic advantage. The maximum mean grain yield 

of (5.91 t/ha) was recorded from the plots where two hand 

weeding at 20 and 40 DAT was performed and was statistically 

at par with the mean grain yield obtained under different weed 

management practices i.e butachlor @ 1.5 kg a.i/ha as pre-em 

fb azimsulfuron @ 25.0 g a.i/ha POE(5.51t/ha), bispyribac 12.5 

g a.i/ha + azimsulfuron 12.5 g a.i/ha POE (5.56t/ha), butachlor 

@ 1.5 kg a.i/ha as pre-em fb ethoxysulfuron @ 15 g a.i/ha as 

POE(5.65 t/ha) and the grain yield obtained these were 

significantly superior to the grain yield obtained under rest of 

the weed management practices. Significantly lowest mean 

grain yield of (4.21 t/ha) was obtained from weedy check 

treatment. The highest net return Rs, 43083/ha was noted in 

treatment butachlor @ 1.5 kg a.i/ha as pre-em fb ethoxysulfuron 

@ 15 g a.i/ha as POE and maximum benefit: cost ratio of 1.92 

was obtained in treatment bispyribac 12.5 g a.i/ha + 

azimsulfuron 12.5 g a.i/ha POE and lower value of net return 

Rs, 28488/ha and benefit: cost ratio of (1.29) were recorded 

under treatment weedy check. 

Key words Grain yield, Herbicides, Rice, Weed density, Economics. 

Rice (Oryza sativa) is one of the most important cereals 

crops grown over diverse environment and geographical 

ranges for human food, feed, fodder and raw materials for 

industries. In Bihar, total area under this crop is 33.37 lakhs 

ha, producing 83.05 metric tons and with average productivity 

of 2489 kg/ha (Anon., 2011-12). Transplanted rice is infested 

by heterogeneous types of weed flora consisting of grassy, 

broadleaf weeds and sedges causing yield reduction up to 

76% (Singh, et. al., 2004). About 60% of the weeds emerge 

during 7-30 day after transplanting and strongly compete with 

rice plants up to maximum tillering stage (Saha and Rao, 2010). 

Therefore, timely weed control at early stage is imperative for 

realizing desired level of productivity from transplanted rice. 

The use of herbicides offers selective and economic control 

of weeds right from the beginning, giving crop an advantage 

of good start and competitive superiority. A number of preemergence 

herbicides like butachlor, pretilachlor, anilophos 

etc. have been recommended for the control of early flushes 

of grassy weeds in transplanted rice field. The intensive use 

of such herbicides year after year has resulted in herbicide 

resistance problems and consequently, management of weeds 

is becoming increasingly more difficult and complex. 

Moreover, continuous use of these herbicides leads to a shift 

of weed flora from grassy to non grassy broadleaf weeds and 

annual sedges (Rajkhowa, et al., 2006). 

Hence, keeping in view the above fact there is a need to 

reduce the evolution of herbicide resistance in weeds and the 

shift towards problematic weeds by avoiding the continuous 

use of a single herbicide or herbicides with similar mechanisms 

of action. Given that herbicide resistance is likely to increase, 

research is needed to evaluate the performance of different 

herbicides, which can provide effective weed control under 

transplanted rice 


A field experiment was carried out during rainy seasons 

(kharif) of 2011 and 2012 at Bihar Agricultural University, 

Sabour, Bihar campus in a randomized block design with three 

replications. The treatments comprised 12 weed management 

practices viz., Un-weeded check, Hand Weeding (HW) at 20 

and 40 Days After Transplanting(DAT), butachlor @ 1.5 kg 

a.i./ha as pre-emergence, butachlor @1.5 kg a.i/ha preemergence 

followed by (fb) 2,4-D easter 0.5 kg a.i./ha post 

emergence, bispyribac sodium @ 20 g a.i/ha as post emergence, 

bispyribac sodium @ 25 g a.i/ha as post emergence, bispyribac 

sodium @ 30 g a.i/ha as post emergence, butachlor @ 1.5 kg 

a.i/ha as pre-emergence fb azimsulfuron @ 25.0 g a.i/ha post 

emergence, penoxsulam @ 22.5 g a.i/ha as post emergence, 

bispyribac 12.5 g a.i/ha + azimsulfuron 12.5 g a.i/ha post 

emergence, oxadiargyl @ 90 g a.i/ha as pre-emergence., 

butachlor @ 1.5 kg a.i/ha as pre-emergence fb ethoxysulfuron 

@ 15 g a.i/ha as post emergence. Twenty one days old

KUMAR et. al., : Effectiveness and Economics of Integrated Weed Management in Transplanted Rice (Oryza sativa) 213 

Table 1. Effect of weed management practices on growth, yield attributes of Rice (Pooled data of two years) 

Treatments Dose (g ai/ha) Stage of application 

(DAS) 

Plant height 

(cm) 

Panicle/m 2 Panicle 

length (cm) 

Grain /panicle Test 

weight(g) 

Weedy check - - 108.2 158 20.1 99 27.21 

HW at 20 and 40 DAT - 20fb40 116.2 194 25.2 119 28.85 

Butachlor 1500 3 110.6 164 21.6 106 28.51 

Butachlor fb 2,4-D Easter 1500fb500 3fb20 114.3 182 22.7 114 28.66 

Bispyribac sodium 20 20 112.5 168 22.1 109 28.61 

Bispyribac sodium 25 20 113.4 178 22.3 111 28.62 

Bispyribac Sodium 30 20 113.6 181 22.5 111 28.62 

Butachlor fb Azimsulfuron 1500fb25 3fb20 114.5 189 23.2 112 28.80 

Penoxsulam 22.5 20 113.0 174 22.0 107 28.50 

Bispyribac + Azimsulfuron 12.5+12.5 20 115.4 191 24.3 116 28.83 

Oxadiargyl 90 3 111.3 165 22.2 110 28.68 

Butachlor fb Ethoxysulfuron 1500fb15 3fb20 115.6 192 24.4 117 28.84 

SEm± - - 3.4 4.69 0.73 1.94 1.22 

CD (P=0.05) - - 11.34 17.57 2.32 6.51 NS 

seedlings of the test variety ‘Rajendra Bhagwati’ was 

transplanted at 20x10cm row spacing on 25 and 28 June, 2011 

and 2012 respectively. In Rice, half recommended dose of N 

(60 Kg/ha) and full dose of P, K and Zn were applied before 

transplanting at final land preparation and the remaining 

nitrogen (60kg/ha) were applied in two splits dose, half at 

active tillering and the rest half at panicle initiation stage. All 

the other recommended agronomic and plant protection 

measures were adopted to raise the crop. Pre-emergence and 

Post-emergence herbicides were applied in saturated soil 

moisture as per the protocol of application time using 

knapsack sprayer fitted with flat fan nozzle at spray volume of 

600 liters/ha. The experimental soil was sandy-loam in texture 

with pH 7.1. The organic carbon, electrical conductivity, 

available nitrogen, phosphorus and potash were 0.59%, 

0.106ds/m, 271.3, 16.41 and 289.36 kg/ha, respectively. The 

rainfall received during the crop season of respective years 

was 1202 mm and 670 mm respectively. The data on weed 

population, weed dry weight and weed control efficiency were 

recorded at different stages of rice crop with the help of a 

quadrate (0.5mx0.5m) at three places and then converted into 

per m 2 . These were subjected to squire root transformation to 

normalize their distribution. Weeds were cuts at ground level, 

washed with tap water. The biomass was determined after 

drying the samples in an oven at 70 0 C for 72 hrs. Weed control 

efficiency (WCE) was calculated by using the formula: WCE 

%=( weed biomass weedy check-weed biomass in managed 

treatment)/weed biomass in weedy check×100.Grain yield of 

rice along with other yield attributing characters like effective 

panicles/m 2 were recorded at harvest. Grain yield was converted 

to t ha -1 at 14% moisture content. Cost of cultivation and gross 

return were calculated on the basis of prevailing market prices 

of different inputs and produces, respectively. 


Weed flora 

The most predominant weed species present in the 

experimental site were Cynodon dactylon (L) Pers, Cyperus 

rotundus (L)., Cyperus iria (L), Cyperus difformis 

(L).,Digitaria sanguinalis (L) ., Fimbristylis miliacea (L) 

Vaha., Eclipta alba (L).,Echinochloa colona ( L).link, 

Echinochloa crus-galli (L) P. Beauv, Marsilea quadrifoliata 

(L).,Eleusine indica , (L) Phyllanthus niruri (L), Euphorbia 

hirta (L), Amaranthus viridis (L), and Commelina 

benghalensis (L). The combination of grasses, sedges and 

broad leaf weeds in weedy check plot were 20, 32, and 48% 

respectively. Emergence of broad leaf weeds was noticed 

earlier than of sedges and grasses. 

Weed density and weed dry weight were higher at 30 

days after transplanting (DAT) then 60 DAT. This was probably 

because of death of some of the weeds like), Cyperus difformis, 

Echinochloa crus-galli,, Marsilea quadrifoliata and 

Commelina benghalensis and the shading effect of the tall 

weeds like Echinochloa crus-galli and crop plant on short 

nature weeds. At both the stages of observation unweedy 

check recorded significantly higher weed population and weed 

dry weight than any other treatment (Table 2). Two hands 

weeding at 20 and 40 DAT recorded the minimum weed 

population and dry weight and the highest weed control 

efficiency at both the stages. Among the herbicidal treatments, 

application of butachlor @ 1.5 kg a.i/ha as pre-em fb 

azimsulfuron @ 25.0 g a.i/ha POE and bispyribac 12.5 g a.i/ha 

+ azimsulfuron 12.5 g a.i/ha POE, recorded the highest grain 

yield which was on par with application of butachlor @ 1.5 kg 

a.i/ha as pre-em fb ethoxysulfuron @ 15 g a.i/ha as POE and 

these three treatments were comparable with 2 hand weedings. 

The reduction weed density and dry weight may be attributed 

to broad spectrum and season long weed control properties 

exhibited by the herbicide mixture or sequential application. 

Unweeded control plots recorded the highest weed population 

and dry weight and lowest weed control efficiency. Among 

the herbicides treatments, application of oxadiargyl @ 90 g 

a.i/ha as Pre–em. recorded higher weed population dry weight 

and lower weed control efficiency, indicating its 

ineffectiveness. Application of butachlor @1.5 kg a.i/ha pre-


Table 2. 

Effects of weed management practices on weed biomass weed density and weed control efficiency in Rice (Pooled data 

of two years) 

Treatment Dose(g/ha) Stage of Weed Population 

application (No/m2) at 30 

(DAS) DAS 

Weed Population 

(No/m2) at 

60 DAS 

Data subjected to sq.root (“x+0.5) transformation. Figures in parentheses are Transformed value. 

Dry Wt. of Weed 

(g/m2) at 30 DAS 

Dry Wt. of 

Weed (g/m2) 

at 60 DAS 

Weed control 

Efficiency 

(%) at 60 DAS 

Weedy check - - 151(12.78) 200(14.64) 13.1(4.11) 18.1(4.75)) - 

HW at 20 and 40 DAT - 20fb40 18(4.74) 24(5.39) 1.1(1.54 1.2(1.59) 93.3 

Butachlor 1500 3 24(5.39) 48(7.42) 1.2(1.59) 2.4(2.04) 86.7 

Butachlor fb 2,4-D Easter 1500fb500 3fb20 20(4.97) 43(7.05) 1.1(1.54) 2.3(2.01) 87.2 

Bispyribac sodium 20 20 46(7.28) 39(6.74) 2.2(1.98) 2.3(2.01) 87.2 

Bispyribac sodium 25 20 44(7.13) 36(6.5) 2.3(2.01) 2.3(2.01) 87.2 

Bispyribac Sodium 30 20 42(6.98) 32(6.15) 2.0(1.91) 2.1(1.94) 88.3 

Butachlor fb Azimsulfuron 1500fb25 3fb20 22(5.19) 28(5.79) 1.2(1.59) 1.4(1.68) 92.2 

Penoxsulam 22.5 20 55(7.91) 34(6.33) 2.6(2.11) 2.1(1.94) 88.3 

Bispyribac + Azimsulfuron 12.5+12.5 20 28(5.79) 30(5.97) 1.7(1.80) 1.4(1.68) 92.2 

Oxadiargyl 90 3 36(6.5) 54(7.84) 2.1(1.94) 3.8(2.44) 79.0 

Butachlor fb Ethoxysulfuron 1500fb15 3fb20 27(5.69) 35(6.41) 1.7(1.80) 2.2(1.98) 87.8 

SEm± - - 4.24 3.4 0.32 0.35 - 

CD (P=0.05) - - 9.1 7.1 0.66 0.74 - 

Table 3. 

Effect of weed management practices on grain yield, gross return, net return and benefit: cost ratio of Rice (Pooled 

data of two years) 

Treatment 

Dose 

(g/ha) 

Stage of application 

(DAS) 

Grain 

Yield( t/ha) 

Gross return 

(Rs./ha) 

Net return 

(Rs./ha) 

Benefit : cost 

ratio (Rs.) 

Weedy check - - 4.21 50520 28488 1.29 

HW at 20 and 40 DAT - 20fb40 5.91 70920 42888 1.52 

Butachlor 1500 3 4.63 55560 31848 1.34 

Butachlor fb 2,4-D Easter 1500fb500 3fb20 5.30 63600 39568 1.64 

Bispyribac sodium 20 20 4.93 59160 36128 1.56 

Bispyribac sodium 25 20 5.14 61680 38148 1.62 

Bispyribac Sodium 30 20 5.27 63240 39208 1.63 

Butachlor fb Azimsulfuron 1500fb25 3fb20 5.51 66120 42108 1.75 

Penoxsulam 22.5 20 5.04 60480 37448 1.62 

Bispyribac + Azimsulfuron 12.5+12.5 20 5.56 66720 43938 1.92 

Oxadiargyl 90 3 4.85 58200 35388 1.55 

Butachlor fb Ethoxysulfuron 1500fb15 3fb20 5.65 67800 43083 1.74 

SEm± - - 0.27 907.3 696.7 0.063 

CD (P=0.05) - - 0.83 1981.8 1445.1 0.131 

em fb 2,4-D easter 0.5 kg a.i./ha POE , bispyribac sodium @ 20 

g a.i/ha as POE , bispyribac sodium @ 25 g a.i/ha as POE and 

bispyribac sodium @ 30 g a.i/ha as POE were also equally 

effective. 

Effect on rice 

All the weed control treatment combinations 

significantly reduced the weeds as compared to weedy check 

and recorded higher grain yield of rice. Among weed control 

measures, the hand weeding twice, butachlor @ 1.5 kg a.i/ha 

as pre-em fb azimsulfuron @ 25.0 g a.i/ha POE, bispyribac 12.5 

g a.i/ha + azimsulfuron 12.5 g a.i/ha POE and butachlor @ 1.5 

kg a.i/ha as pre-em fb ethoxysulfuron @ 15 g a.i/ha as POE 

increased the grain yield by 40.38, 30.95,32.0 and 34.20% 

respectively, over the unweedy check. The increased grain 

yield in these treatments were owing to reduced weed density, 

weed dry weight and better weed control efficiency and higher 

panicles/unit area (Table 2). This might due to better control 

of broad leaf and sedges which were prevalent in the rice field 

during both the year.. The minimum yield and yield attributes 

in unweeded check were the result of severe weed competition 

by the uncontrolled weed growth. Nawal, et. al., 2002 reported 

increased grain yield by herbicide mixture and sequential 

herbicide application respectively. There was no phytotoxic 

effect of any herbicides at any of doses applied alone or tank 

mixture in transplanted rice crop. Application of the hand 

weedings twice, butachlor @ 1.5 kg a.i/ha as pre-em fb 

azimsulfuron @ 25.0 g a.i/ha POE, bispyribac 12.5 g a.i/ha + 

azimsulfuron 12.5 g a.i/ha POE and butachlor @ 1.5 kg a.i/ha 

as Pre-em fb ethoxysulfuron @ 15 g a.i/ha as POE recorded 

the highest net return and maximum benefit: cost ratio (Table 

3). This may be due to high weed control efficiency and higher 

grain yield obtained owing to application of effective herbicide 

dose and combination. Thus application of butachlor @ 1.5

KUMAR et. al., : Effectiveness and Economics of Integrated Weed Management in Transplanted Rice (Oryza sativa) 215 

kg a.i/ha as pre-em fb azimsulfuron @ 25.0 g a.i/ha POE, 

bispyribac 12.5 g a.i/ha + azimsulfuron 12.5 g a.i/ha POE and 

butachlor @ 1.5 kg a.i/ha as pre-em fb ethoxysulfuron @ 15 g 

a.i/ha as POE proved more effective in checking the weed 

population and their growth and increasing the grain yield in 

transplanted rice. All the weed control methods resulted 

significant increase in grain and biological yield over weedy 

check. 


Anonymous, 2012. Statistcs Department of Agriculture, Govt. of Bihar 

(2011-2012) 

Nawal, Sandeep, Singh, Samar, Panwar, K.S. andMalik, R.K. 2002. 

Performance of acetochlor and anilofos + ethoxysulfuron for weed 

control in transplanted rice (Oryza sativa). Indian Journal of 

Agronomy, 47(1):67-71. 

Rajkhowa, D.J., Borah.N, Barua.I.C and Deka, N.C.2006. Effect of 

pyrazosulfuron ethyl on weed and productivity of transplanted rice 

during rainy season. Indian Journal of Weed science, 38(1& 2): 25-28. 

Saha, Sanjoy and Rao. K.S. 2010.Efficacy of metsulfuron methyl for 

controlling broadleaf weeds in transplanted rice (Oryza sativa) under 

rainfed shallow lowland .Indian Journal of Agricultural Sciences, 

80 (6):522-26. 

Singh, V.P., Singh Govindra and Singh Mahendra. 2004. Effect of 

fenoxaprop-p-ethyl in transplanted rice and associated weeds. 

Indian Journal of Weed Science, 36 (3 & 4): 190–192. 



Biology of Mango – Hopper, Amritodus atkinsoni (Leth.) (Jassidae : Hemiptera) in 

Agro – ecosystem of Manipur 

M. BHUBANESHWARI DEVI 

P. G. Department of Zoology, Laboratory of Entomology, D.M. College of Science, Imphal 

e-mail: aalalplantpathology@gmail.com 

ABSTRACTS 

The mango-hopper, Amritodus atkinsoni (Leth.) (Jassidae : 

Hemiptera) is one of the most serious pests of mango trees in 

Manipur. The maximum infestation is seen during the 

(February and March) flowering season when the temperature 

is 28 0 C (Max.), 25 0 C (Min.); R.H. 85% (Max.), and 75% (Min.); 

Rainfall almost nil. During other months, they are very few in 

number and confine themselves in cracks and crevices of the 

bark of trees. The life cycle continues from February / March 

to April / May. The eggs are laid on rachis of inflorescence, 

having incubation period from 8 to 12 days. There are five 

larval instars with four moultings, the total nymphal period 

including five instars lasted from 15 to 20 days and the total 

life cycle from egg to adult ranged from 25 to 28 days under the 

laboratory conditions. 

Key words Biology, Amritodus atkinsoni, mango pest, Manipur. 

Mango, the king of the fruits, Mangifera indica Linn. 

(Anacardiaceae) is known due to its attractive colour, odour 

and a nice delicacy. It is the most important fruit crop of 

Manipur. But this important fruit of mango is infested by 

several pests. It has been reported that over 175 species of 

insects infests damage mango regularly (Fletcher, 1920, Veva, 

1969 and Nayar, et.al., 1976). These pests damage several 

parts of the mango tree. Reddy 1968 reported that extent of 

damage and the loss in yield of mangoes due to this pest 

varied in the different areas while Wadhi and Batra 1964 

reported 25% – 60% loss in general. Anufriew 1970 reported 

that the mango hopper Amritodus atkinsoni is one of the 

most serious pests of mango trees in Punjab, Bihar, southern 

India and Gujarat. The life cycle of the pest is significant to 

make effective IPM measures to suppress the pest 

manifestation in Manipur. 


The mango hoppers were collected from the field and 

kept in glass rearing jars/Plastic rearing jars covered with fine 

muslin cloth. Inside the rearing jars small plastic boxes fitted 

with water containing fresh mango spikes were kept for egg 

laying. These spikes were changed twice daily to get the 

nymph enough pest food. The observations were carried out 

at room temperature (25±3 0 C and 80±5% R.H.). 


Life history : 

Egg: 

The eggs were laid singly but embedded in the tissue 

and only blunt end was visible. The eggs were smooth, creamy 

white and oblong in shape. It measure on an average 0.70 ± 

0.07 mm in length and 0.50 ± 0.02 mm in diameter. They are 

pointed at one end and blunt at the other end. The incubation 

period ranged from 8 – 12 days. Reddy, 1968 observed that 

the egg stage occupied 7 – 10 days while according to Patel 

and Talgiri 1950, it ranged from 8 – 10 days. Patel, et.al., 1975 

reported that the incubation period ranged from 7 – 9 days. 

First instar larva: 

There are five nymphal instars within a period of 17 – 20 

days. The nymphs are voracious eaters and excreted semiliquid 

sticky substances, the honey dew. 

The newly hatched nymph is very delicate and pale 

yellow in colour. It conspicuous bulging red compound eyes. 

It measured on an average 1.04 ± 0.02 mm in length and 0.49 ± 

0.05 mm in breadth. The duration of this instar ranged from 4 

– 5 days. Patel, et.al., 1975 reported that the first instar ranged 

from 3 – 4 days. 

Second instar larava: 

The first instar larva after 4 – 5 days moulting changed 

into second instar larva. It is yellowish in colour but later on 

changed into dark brown. The head is relatively larger than 

the remaining parts of the body. It measured on an average of 

2.0 ± 0.03 mm in length and 0.60 ± 0.04 mm in breadth. The 

duration of the second instar larva ranged from 4 – 6 days 

(table 1). Patel et.al (1975) reported the duration of the second 

instar larva from 3 – 4 days. 

Third instar larva: 

The second instar larva after 3 – 4 days, metamorphosed 

into the third instar larva. It is of yellowish colour at first but 

the dorsal side is dark brown whereas the ventral side is pale 

yellow in colour. The three notum of the thorax are well 

developed and the pronotum overlapped the head. The 

rudiments of two pairs of wings appeared on mesonotum and 

metanotum. Abdominal segments are well differentiated. It

DEVI : Biology of mango – hopper, Amritodus atkinsoni (Leth.) (Jassidae : Hemiptera) in agro – ecosystem of Manipur 217 

Table 1. 

Measurement of the different instars of Amritodus 

atkinsoni (Leth.) in mm 

Stage No. of insects Length (mm) Breadth (mm) 

Egg 

1 st instar 

2 nd instar 

3 rd instar 

4 th instar 

5 th instar 

10 

10 

10 

10 

10 

10 

0.70 ± 0.07 

1.04 ± 0.02 

2.0 ± 0.03 

2.4 ± 0.03 

3.2 ± 0.05 

5.0 ± 0.02 

0.50 ± 0.02 

0.49 ± 0.05 

0.60 ± 0.04 

0.80 ± 0.03 

0.120 ± 0.02 

0.141 ±0.05 

Adult: 

The adults are cream coloured after just emergence but 

later on changed to brown dorsally and pale yellowish 

ventrally. The adult survived 4 – 5 days on natural food under 

laboratory condition. The females are longer than the males. 

The female adult measured on an average of 4.80 ± 0.03 mm in 

length whereas the male measured on an average of 4.51 ± 

Table 2. Duration of different stages of Amritodus atkinsoni (Leth.) 

1 st instar 2 nd instar 3 rd instar 4 th instar 5 th instar 

Dates No. of days Dates No. of days Dates No. of days Dates No. of days Dates No. of days 

10 th – 13 th March 4 13 th – 17 th March 4 17 th – 22 nd March 5 22 nd – 27 th March 5 27 th – 31 st March 4 

12 th – 14 th March 3 14 th – 18 th March 4 18 th – 21 st March 3 21 st – 25 th March 4 25 th – 29 th March 4 

17 th – 20 th March 4 20 th – 23 rd march 3 23 rd – 27 th March 4 27 th – 30 th March 3 30 th – 2 nd April 3 

measured on an average 2.4 ± 0.03 mm in length and 0.80 ± 

0.03 mm in breadth. The duration of the third instar larva ranged 

from 4 – 5 days. Patel, et. al., 1975 reported the duration of the 

third instar larva 3 – 4 days. 

Fourth instar larva: 

The fourth instar larva is also of yellow colour in the 

beginning but changed to dark brown later. The head is 

conspicuously bulging when compared with thorax. The 

compound eyes are dark black in colour. The larvae are more 

active and moved here and there on the inflorescence, buds 

and leaves of mango. They shared diagonal walking 

movements. The wing pads are enlarged and distinct. The 

sizes are differentiated from this stage onwards. It measured 

on an average of 3.2 ± 0.05 mm in length and 0.12 ± 0.02 mm in 

breadth. The duration of the fourth instar larva ranged from 4 

– 5 days (Table 1). Patel, et.al., also observed that the variation 

of 3 – 4 days in this stage. 

Fifth instar larva: 

This fifth instar larva is pale yellow in colour but later on 

the dorsal side changed to dark grey. The wing pads are very 

large and very active. They move in a diagonal way. The size 

of the body is little increase than the previous one. It measured 

on an average 5.0 ± 0.02 mm in length and 0.14 ± 0.05 mm in 

breadth. The duration of this stage ranged from 3 – 4 days but 

Patel, et.al., 1925 observed 2 – 3 days in this experiment. 

Thus the total larval period ranged from 17 – 20 days in 

the climate condition of Manipur. Patel and Talgiri, 1950 

observed the variation from 15 – 17 days. 

0.05 mm in length. The present observations are at par with 

Patel, et.al., 1975 who reported the female’s body length 4.86 

mm and the male’s body length 4.50 mm. 


The author is grateful to the Principal, D.M. College of 

Science, Imphal and Head of Department of Zoology, D.M. 

College of Science, Imphal for giving me laboratory facilities. 

The author remains thanks to the UGC for giving me financial 

assistance. 


Anufrieu, G.A. 1970. Description of a new genus, Amritodus for Idiocerus 

atkinsoni leth. From India (Hemiptera : Cicadellidae). J. nat.Hist., 

4: 375 – 376. 

Fletcher, B.T. 1970. Fruit trees. Report Proc. 2 nd . Ent.Mtg. Pusa (Bihar) 

February. Calcutta (9,43,94,278). 

Nayar, K.K.; T.N. Ananthakryshnan and B.V.David 1976. General and 

Applied Entomology. Tata Mc Graw Hill Publishing Co. Ltd. New 

Delhi pp. 589. 

Patel, G.A. and Talgiri, G.M. 1950. Crop pests and how to fight them. 

Government of Bombay, pp. 126–145. 

Patel, R.K., Patel, S.R., and Shah. A.H., 1975. Biology of mango 

hopper, Amritodus atkinsoni (Leth.) (Jassidae : Hemiptera) in south 

Gujarat. Indian J. Ent., 37(2): 150–153. 

Reddy, D.B. 1968. Plant Protection in India. Allied Publications, New 

Delhi. pp. 238-239. 

Wadhi, S.R. and Batra, H.N. 1964. Pests of tropical and sub tropical 

fruit trees. In: Entomology in India, (ed. N.C. Pant). The 

Entomological Society of India. New Delhi, 227–260 pp. 

Veva, E.J. 1969c. Know your crop, its pest problem and control. Mango 

Pesticides. 3(12): 21 – 31. 


Trends in Biosciences 6 (2): 218, 2013 

SHORT COMMUNICATION 

A New Blight Disease of Rice Caused by Curvularia sp. from U.P. 

KAMALUDDEEN 1 , SOBITA SIMON AND ABHILASHA A. LAL 

Department of Plant Protection, Sam Higginbottom Institute of Agriculture, Technology & Sciences ,(Formerly 

(Deemed-to-be-University), Allahabad - 211 007 (U.P.) India 

e-mail : kamaluddinpatho@gmail.com 1 

Rice is the most important cereal crop grown all over 

the world. During cultivation of rice variety Pant-12, at the 

tillering stage a new disease was observed at central field 

of Sam Higginbottom Institute of Agriculture, Technology 

& Sciences Allahabad. The rice plants were cultivated 

using standard agronomic 

practices (Singh, 2005). 

The first symptoms 

appeared on the leaves as 

elliptical brown spots which 

increased in size. Gradually 

the colour of the spots 

changed to brownish black. 

The length of spots ranged 

from 0.2 to 1cm (Fig.1). Later 

the spots appeared on leaf 

sheath which were dark 

brown initially with yellow 

margin and the diseased 

sheath became yellow and 

blighted. Initially these 

spots were similar to the 

ones on the leaf but later 

covered the whole leaf 

sheath (Fig.2). Gradually the 

disease spread to the 

kernels. Glumes were 

discoloured and in severe 

infection, the rice kernel 

showed black discoloration 

(Fig. 3). 

M i c r o - s c o p i c a l 

examination revealed the 

presence of conidia of the 

fungus. The conidia were 

Fig.1. Disease symptoms 

on the leaves 

Fig. 2. Disease symptoms on 

the leaf sheath 

Fig. 3. Disease symptoms on 

the kernels 

four celled and generally curved, with two middle cells 

darker than the paler end cells. Conidiophores were 

branched, septate and dark in colour. 

Isolation of the fungus from the infected parts was 

carried out on PDA (Potato Dextrose Agar) plates following 

the standard methodology (Aneja, 2003). After two-three 

days whitish mycelial growth appeared which gradually 

became greyish black in colour (Fig.4). 

Microscopical examination of fungal culture again 

showed the presence of conidia and conidiophores typical 

of Curvularia (Mehrotra and Aneja, 2008). The fungus was 

therefore identified as Curvularia sp. (Fig.5). 

Fig. 4. Pure culture 

of Curvularia sp. 

Perusal of literature revealed that incidence of 

Curvularia sp. have been reported on rice grains by Rao 

and Salam 1954 from Hyderabad. This seems to be first 

report of a new blight disease of rice from India. 


Fig. 5. Conidiophores & conidia 

of Curvularia 

Aneja, K. R. 2003. Experiment in Microbiology, Plant Pathology and 

Biotechnology. New Age International (P) Limited, Publishers. New 

Delhi. pp. 439. 

Mehrotra, R. S. and Aneja K.R. 2008. An Introduction to Mycology. 

New Age International (P) Limited, Publishers. New Delhi. pp. 

599. 

Rao, P. N. and Salam, M.A. 1954. Curvularia sp. from discolored 

grains from Hyderabad. Journal Ind. Bot. Soc., 33: 268-271. 

Singh, S. S. 2005. Crop Management. Kalyani Publishers. Ludhiana. 

pp. 50. 



SHORT COMMUNICATION 

Exploring Precision Agriculture Approaches for Insect Pest Management 

1 

PRASHANT KUMAR AND 2 ASHUTOSH KUMAR 

1 

Department of Vegetable Science Punjab Agriculture University (PAU), Ludhiana 

2 

Department of Botany PAU, Ludhiana 

e-mail: prashantpau@rediffmail.com 1 

New tools are needed for the monitoring of insect pests 

on a small spatial scale (Fleischer, et. al., 1999) for the delivery 

of variable rates of insect pest management other than 

pesticides, such as mass release of biological control agents, 

and for geo-statistical analyses to detect spatial relations 

between variables in the environment (Liebhold, et. al., 1993). 

Goal of precision agriculture is to vary management inputs in 

different parts of the field so that crop response is optimised. 

The efficacy of pest control measures could be improved and 

adverse environmental impact be minimised by considering 

the spatial variability in pest occurrence and their habitat 

(Lan, et. al., 2010). The results are used to illustrate how these 

new tools could be used to optimally arrange crop plants in 

order to reduce insect pest-inflicted damage. 

The main tools used in precision agriculture for insect 

pest management mainly includes yield monitors (YM), sensors 

used to monitor yield during the harvest in order to quantify 

yields across the field, variable rate technologies (VRT) that 

are mounted on application equipment such as sprayers to 

control their delivery rates (Ravi and Jagadeesha, 2002). Global 

positioning system (GPS), a satellite-based locating system 

which identifies an earth-based position using longitude and 

latitude. Geographic information system (GIS), a 

data-management system designed to store spatial data and 

create variable-intensity maps. These allow the delivery of 

variable levels of a specific management practice to various 

parts of the field and in a single field operation. For precision 

insect-pest control, these tools should be combined with a 

decision-support system. 

Remote sensing (RS) is being used with GPS, GIS, and 

VRT to ultimately help farmers maximize the economic and 

environmental benefits of crop pest management 

through precision agriculture (Lan, et. al., 2009). In the past, 

pests were hand-picked with the ultimate in precision pest 

control. For many years integrated pest management (IPM) 

has been the approach of choice for pest control. Yet IPM 

lacks the spatial component so central to precision agriculture 

(Fleischer, et. al., 1999). Precision agriculture, because it has 

been designed for use in crop production, must be adapted 

for use in insect pest management. Various authors have 

discussed the application of remote sensing to insect pest 

monitoring (Pathak and Dhaliwal, 1985) and plant protection 

in general (Hatfield and Pinter, 1993). In spite of this, the 

clustered distribution of insect pests does make them suitable 

for this type of management. RS may be advantageous as it 

can detect infested hot-spots by monitoring either the health 

of the plants or visible by-products of infestations, such as 

defoliation and sooty mould-contaminated honeydew that is 

secreted by homopteran pests (Hart and Myers, 1968, Chaing, 

et. al., 1976). Detection of consistent patterns of insect pest 

infestation over several years will enable the preparation of 

multi-year management maps that could be used to apply preemptive 

control measures (Blackmore, 2000). Data collected 

through RS are transferred to a GIS and used to create, for 

example, maps of defoliation attributable to a pest such as the 

Gypsy moth. If indeed particular areas of the forest are seen 

to be defoliated repeatedly over several years, control 

measures could be applied to these susceptible areas before 

insect pest populations reach damaging levels (Liebhold, 

et. al., 1998). GIS is also highly suitable tool for detecting 

such consistent infestation patterns. GIS and GPS-linked data 

collection could be used to optimally arrange crops in an area. 

The small size, cryptic nature, rapid change in spatial 

occurrence and dynamic infestation pattern characteristic of 

insect pest present exceptional challenges. Precision 

agriculture uses new technologies to provide us with a unique 

opportunity to add a spatial dimension to our pest control 

measures. Although the cost of these technologies is currently 

prohibitive, they may become standard farm equipment in the 

future. Small farmers could use precision agriculture 

technologies by a cooperative system to urgently deal with 

some area wide pest management issues. However, sitespecific 

control of insect pest may be possible based on pest 

biology and parameters correlated with pest infestation. 

Finally, precision agriculture methodologies, such as the GPS, 

RS and GIS, may be used for area-wide management of insect 

pests through farm landscaping. Thus, biological 

characteristics of insect pest infestation in a particular system, 

together with economic criteria, will point to the most effective 

approach. 


Blackmore, B. S. 2000. The interpretation of trends from multiple 

yield maps. Computers and Electronics in Agriculture, 26: 37–51. 

Chaing, H. C., Meyer, M. P. and Jensen, M. S. 1976. Armyworm 

defoliation in corn as seen on IR aerial photographs. Entomologia 

Experimentalis et Applicata, 20: 301–03.


Fleischer, S. J., Blom, P. E. and Weisz, R. 1999 Sampling in precision 

IPM : When the objective is a map. Phytopathology, 89: 1112–18. 

Hart W. G. and Myers V. I. 1968 Infrared aerial colour photography for 

detection of populations of brown soft scale in citrus groves. Journal 

of Economic Entomology, 61: 617–24. 

Hatfield, J. L. and Pinter, P. J. 1993. Remote sensing for crop protection. 

Crop Protection, 12: 403–13. 

Landis D A and Marino P C (1999) Landscape structure and extrafield 

processes: impact on management of pests and beneficials. In 

Handbook of Pest Management. Marcel Dekker, New York pp. 

74–104. 

Lan, Y. Thomson, S. J., Huang, Y. Hoffmann, W. C., Zhang, H. 2010. 

Current status and future directions of precision aerial application 

for site speciûc crop management in the USA. Computers and 

Electronics in Agriculture, 74: 34–38 

Liebhold, A. M., Rossi, R. E. and Kemp, W. P. 1993. Geostatistics and 

geographic information systems in applied insect ecology. Annual 

Review of Entomology, 38: 303–27. 

Liebhold, A., Luzaader, E., Reardon, R. Roberts, A. Ravlin, F. W., Sharov, 

A. and Zhou, G. 1998. Forecasting Gypsy moth defoliation with a 

Geographical Information System. Journal of Economic 

Entomology, 91: 464–72. 

Pathak, M. D. and Dhaliwal, G. S. 1985. Remote sensing and monitoring 

of insect pest problems in rice. In: Application of Remote Sensing 

for Rice Production pp: 77–87. 

Ravi, N. and Jagadeesha, C. J. 2002. Precision Agriculture, Training 

course on Remote Sensing and GIS. Applications in Agriculture, 

RRSSC-Bangalore, pp: 225-28. 


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Research Papers: Each full length of research papers should be covered within 1500 words including tables and illustrations. 

Short communication should be within 175 words including tables, figures and references, in case of exceeding the limit, payment 

has to be made for extra materials by the authors. Correct language is the responsibility of the author. No editing or materials 

changes at the proof stage will be permitted. While short communication will have only title, authors name, address and e-mail 

followed by text and references. In case of full length paper authors should have the following headings. 

Title : The title to be typed in capital and small letters, author names (all capital) and affiliation (capital and small letters with 

italics fonts). Give e-mail address in italic fonts also. Manuscript must confirm to the journal style (see latest issue) 

ABSTRACT : The abstract should indicate the main findings of the papers and typed in bold, single space. It should be not 

more than 150 words. The abstract should be typed before the main text and intended ca. 2cm to the right of it. 

Key words 5-6 key words in italics should be given. 

Tables and figures : Table should be descriptive without and references to the text with heading in bold letters. Each table 

should be typed on a separate sheet. Figures whether line drawing or graph should be of good quality. Legends to figures should 

be given on a separate sheet. Tables and figures should be numbered consequently in Arabic numerals (Table 1., Table 2., Fig.1., 

Fig.2.- 3) can be identified on the back by name (s) of the author(s). 

Introduction : This should be brief and related to aim of the study. The review of the literature should be pertinent to the 

theme of the paper. Extensive review and unnecessary details of earlier work should be avoided. Heading “introduction” should 

not be written. 

MATERIALS AND METHODS : When methods are well known, citation of the standard work is sufficient. All measurement 

should be in metric units. 

RESULTS AND DISCUSSION : The result should be supported by brief and adequate tables, graphs and charts, wherever 

needed. 

LITERATURE CITED : In the text, references should be cited as follows: two authors (Ali, and Pervez, 2005), three or more 

authors (Gaugler, et al., 2001). All references made in the text must be listed under LITERATURE CITED at the end of the text. 

References should be listed alphabetically by the authors, followed by the year of the publication. Journal titles should be cited 

in full and in italics, while for books the place of the publication should precede the name of the publisher, Example Strong, D.R. 

2002. Population of entomopathogenic nematodes in food webs. In: Entomopathogenic Nematology, (ed.Randy Gaugler) CAB 

International, ix + 387. pp. 225-240. 

Fox, P.C. and Atkinson, H.J. 1984. Glucose phosphate isomerase polymorphism in field population of the potato cyst 

nematode, Globedera rostochiensis and G. pallida. Annals of Applied Biology, 104(1):503-506. 

Submission of manuscript : Duplicate copy of manuscript along with soft copy (CD) should be submitted to the Editor in 

Chief, Dr. S.S. Ali, H-1312, VIP Lane, Satyam Vihar, Avas Vikas No. 1, Kalyanpur, Kanpur 208 018, India. The text may sent through 

e-mail to ss_ali@rediffmail.com. Payment must be made through Demand Draft/e-banking payable to State Bank of India, 

Kalyanpur Branch, (Branch code 01962, IFSC : SBIN0001962) Kanpur, U.P. India, in favour of account of “Trends in Bioscience” 

(Account number 31575871348) along with manuscript and send to Dr. S. S. Ali, Editor in Chief. For online subscription of 

journal contact info@indianjournals.com 

Copyright : copyright © of all papers published in Trends in Biosciences, acceptance of manuscript for Trends in Biosciences, 

automatically transfers the copyright to Trends in Biosciences The use of trade name or a propriety product does not constitute 

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may be suitable.

TRENDS IN BIOSCIENCES JOURNAL 6-2 JUNE 2013 EDITION

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