concentration of polysaccharides in spring - vii-th international ...

2 

VII th ARTHROPODS, 18-23 September, 2011, Białka Tatrzańska, Poland 

Faculty of Chemistry 

VII th INTERNATIONAL CONFERENCE 

ON ARTHROPODS: CHEMICAL, 

PHYSIOLOGICAL, BIOTECHNOLOGICAL 

AND ENVIRONMENTAL ASPECTS 

Stefan Kopeć Memorial Conference 

A B S T R A C T S 

September 18-23, 2011 

Białka Tatrzańska 

Poland


Front cover design: Mariola Kuczer 

© Copyright by Faculty of Chemistry, University of Wrocław, 2011 

Abstracts of the conference edited by: 

DANUTA KONOPIŃSKA and MARIOLA KUCZER 

Faculty of Chemistry, University of Wrocław, Poland 

ISBN: 978-83-60043-16-5 

Printed by: 

Wrocławska Drukarnia Naukowa PAN Sp. z o.o. 

ul. Lelewela 4, 53-305 Wrocław 

http://www.wdn.pl 

3

4 


Scientific Committee: 

Danuta Konopińska (Poland) – chair 

Dov Borovsky (USA) 

Arnold De Loof (Belgium) 

Gerd Gäde (South Africa) 

Jadwiga Giebultowicz (USA) 

Graham Goldsworthy (UK) 

Angela B. Lange (Canada) 

Paweł Migula (Poland) 

Ronald J. Nachman (USA) 

Jan Nawrot (Poland) 

Local Organizing Committee: 

Danuta Konopińska 

Stanisław Ignatowicz 

Paweł Migula 

Bogumił Leszczyński 

Jerzy Łukasiak 

Remigiusz Olszak 

Secretariat: 

Mariola Kuczer 

Alicja Kluczyk 

Remigiusz Olszak (Poland) 

Ian Orchard (Canada) 

Grzegorz Rosiński (Poland) 

Frantińek Sehnal (Czech Republic) 

Karel Slama (Czech Republic) 

Guy Smagghe(Belgium) 

EugeniaTęgowska (Poland) 

Czesław Wawrzeńczyk (Poland) 

Dusan Zitnan (Slovakia) 

Andrzej Plech 

Grzegorz Rosiński 

Maryla Szczepanik 

Eugenia Tęgowska 

Czesław Wawrzeńczyk 

Anna Gliszczyńska


5


PLENARY 

LECTURES

7 

VII th ARTHROPODS, 18-23 September, 2011, Białka Tatrzańska, Poland


PL-1 

MIMETIC ANALOGS OF INSECT NEUROPEPTIDES AS 

RATIONAL TOOLS FOR DEVELOPMENT OF NOVEL 

PEST CONTROL STRATEGIES 

8 

Nachman R., Coast G., Kaczmarek K., Zabrocki J., Denlinger D., Smagghe G. 

Southern Plains Agricultural Research Center, U.S. Department of Agriculture, 

College Station, TX 77845, USA 

Insect neuropeptides regulate critical processes and behaviors in insects, 

though they are unsuitable as tools to arthropod endocrinologists and/or as pest 

management agents due to unsuitable biostability and/or bioavailability characteristics. 

Peptidomimetic analogs can overcome these limitations and either over-activate 

or block critical neuropeptide-regulated functions. Biostable agonists of the 

diuretic/myotropic myokinin and tachykinin-related (TRP) classes of neuropeptides 

were developed that match or exceed the potency of natural peptides 

in bioassays/expressed receptors. When fed to aphids they lead to potent induction 

of high mortality, matching or exceeding the potency of several commercial aphicides. 

Unmodified myokinin and TRP peptides are virtually inactive in the aphid assays. 

Highly potent mimetic analogs of the allatostatin class are also discussed. Diapause 

hormone (DH) has been shown to terminate pupal diapause of heliothine species. 

A novel antagonist prevents diapause termination. A potent agonist analog both 

prevents the onset of pupal diapause in heliothine larvae and leads to high mortality, 

apparently affecting both diapause induction and ecdysis. Modified analogs 

demonstrate that the pupal diapause-prevention and mortality effects can be separated. 

These analogs provide leads for the generation of agents capable of disrupting 

diapause (and ecdysis) in economically important lepidopteran pests. 

- STEFAN KOPEĆ MEMORIAL LECTURE -


PL-2 

RNA INTERFERENCE – STUDIES OF GENE 

FUNCTIONS IN ARTHROPODS 

Klaus Hoffmann 1 , Martina Meyering-Vos 1 , Mohatmed Abdel-Latief 2 , Laura Vannini 3 , 

Franziska Wende 1 , Joseph Woodring 1 

1 Department of Animal Ecology I, University of Bayreuth, 95440 Bayreuth, Germany. 

2 Institute of Biology, Free University of Berlin, 12163 Berlin, Germany. 

3 Department of Evolutionary Biology, University of Siena, 53100 Siena, Italy. 

e-mail: klaus.hoffmann@uni-bayreuth.de 

The increasing availability of invertebrate genomes has revealed a large 

number of genes with unknown functions. RNA interference (RNAi) or post 

translational gene silencing has emerged as a powerful tool for sequence-specific 

down-regulation of gene expression thus elucidating in vivo functions of gene 

transcripts in insects and other arthropods. RNAi can unveil the functions of new 

genes, especially in non-model organisms, lead to the discovery of new functions for 

known genes, and find the genes for old functions [Belles X. (2010) Annu. Rev. 

Entomol. 55, 111]. Many but not all arthropods studied so far exhibit a systemic RNAi 

response. Besides its use in functional genomic research, RNAi has also a considerable 

potential for the control of pest species [Huvenne H., Smagghe G. (2010) J. Insect 

Physiol. 56, 227]. 

In gene function studies, the method has mainly relied on the injection of the doublestranded 

RNA (dsRNA), which is not possible for a practical application in pest 

control. However, recent results have shown that dsRNA fed as a diet component is 

also effective, at least in some species. Moreover, expression of the dsRNA directed 

against suitable arthropod target genes in transgenic plants has been shown to give 

protection against pests [Price D.R.G., Gatehouse J.A. (2008) Trends Biotechnol. 26, 

393]. 

After a short review on the current status of RNAi-based experiments in insects [Mito 

T. et al. (2011) Entomol. Sci. 14,1], crustaceans [Kato Y. et al. (2011) Dev. Genes 

Evol. 220, 337], and spiders [Prpic N.M. et al. (2008) Cold Spring Harb. Protoc. 

pdp.prot 5070] I shall present results from our laboratory, where RNAi has been used 

to elucidate the role of allatoregulating neuropeptides (allatotropins, allatostatins) and 

juvenile hormone (JH) synthesis and degradation enzymes, respectivel, in controlling 

haemolymph JH titers, and thereby fertility of females from various “model” and nonmodel” 

insects (Tribolium castaneum, Ceratitis capitata, Gryllus bimaculatus, 

Spodoptera frugiperda, Mastotermes darwiniensis). 

Allatostatins (AS) and allatotropins (AT) also have been shown to affect spontaneous 

contractions of the (fore)gut in a variety of insects [Audsley N., Weaver R.J. (2009). 

Gen. Comp. Endocrinol. 162, 93]. Therefore, we used the RNAi technique to see, 

whether AS, AT and other neuropeptides (e.g. sulfakinins) are associated with the 

regulation of feeding and the secretion of digestive enzymes in crickets and 

armyworms. 

Understanding the molecular details of (neuro)peptide actions in a large spectrum 

of arthropods may certainly help to find genes which should be targeted in RNAimediated 

pest control. 

- PLENARY LECTURES - 9

10 


PL-3 

SMALL RNAS AND TRANSPOSON AND GENE 

REGULATION IN MOSQUITOES 

Arensburger P. 1 , Hice R.H 1 ., Wright J.A 1 ., Craig N. L. 2 , Atkinson P.W. 1 

1Department of Entomology and Institute of Integrative Genome Biology, University 

of California, Riverside, CA 92521, USA, peter.atkinson@ucr.edu 

2Department of Molecular Biology and Genetics and HHMI, Johns Hopkins School 

of Medicine, Baltimore, MD, 21205, USA 

The piRNA pathway is one of three small RNA pathways present in metazoans 

and, based on studies in model organisms, plays a significant role in the regulation 

of transposons in the genome thereby leading to increased genome stability. We report 

on the characterization of the piRNA pathway in the mosquito, Aedes aegypti, and 

insect with a large genome size (1.38 Gb) and a large and diverse transposon 

complement (46%, with the members of most transposon superfamilies present). Ae. 

aegypti also contains an expansion of the piwi genes involved in piRNA biogenesis. 

We find that Ae. aegypti has a very robust piRNA response to transposons however 

many non-transposon gene sequences also appear to be targets of this small RNA 

response, including genes implicated in piRNA biogenesis. We also explored the 

relationship between the abundance of piRNAs directed to transposons and the 

transcription levels of transposon-specific transcripts using RNAseq analysis 

of ovaries and eggs. 

- PLENARY LECTURES -


- PLENARY LECTURES - 11


ORAL 

PRESENTATIONS

13 


14 


O-1 

THE ALLATOTROPIN- LIKE RECEPTOR OF 

TRIBOLIUM CASTANEUM 

Roger Huybrechts 1 , Yoshiki Nagaba 1 , Marleen Lindemans 2 , Heleen Verlinden 3 , 

Kristel Vuerinckx 1 

1 Laboratory of Insect Physiology and Molecular Ethology, KULeuven, Leuven, 

roger.huybrechts@bio.kuleuven.be; nagaba.nagaba@bio.kuleuven.be; 

kristel.vuerinckx@bio.kuleuven.be 

2 Laboratory of Genomics and Proteomics, KUleuven, Leuven, 

marleen.lindemans@bio.kuleuven.be 

3 Molecular Developmental Physiology and Signal Transduction, KULeuven, Leuven, 

heleen.verlinden@bio.kuleuven.be 

In Tribolium 3 genes each encodes a precursor isoform of one and the same 

allatotropin- like peptide. No Manduca like allatotropin peptide has been annotated 

in beetlebase and the ATL peptide is effective in stimulating JH release from corpora 

allata of Tenebrio, making TricaATL a true allatotropin (Abdel-Latief and Hoffman 

(2010) Arch. Insect Biochem. Physiol. 74,135). We succeeded in identifying the 

TricaATL receptor which was formerly annotated as orexin type two receptor. Our 

finding confirms the earlier suggestion of the orexin like receptor being an allatotropin 

receptor in Bombyx ((Yamanaka et al., (2008) Plos one,3,:e3048). Corresponding 

receptors were simultaneously identified as true allatotropin receptor in Manduca 

(Horodyski et al (2011) J.insect biochem.mol.biol.in press) and Aedes (Noriega et al, 

(2011) personal communication). 

Apart from its presence in the head (CC-CA?, frontal ganglion?) Q-PCR indicates that 

this receptor is strongly expressed in the gonadal tract, males always displaying high 

expression in the accessory glands. Northern analysis and Western blotting also 

pinpoint ATL receptor protein presence in the accessory glands but also demonstrates 

immunopositive signal in the testes. Q-PCR further indicates minor expression 

of TricaATL receptor in the intestine but relevant expression in fat body. All together 

our present data about ATL receptor in Tribolium castaneum indicate a role for the AT 

receptor pathway beyond JH synthesis. The mRNA profiling of the ATL peptide 

precursor, indicating ATL neuropeptide expression outside the head in as well 

accessory glands and gonads as in fat body and intestine further substantiate this 

hypothesis. 

- ORAL PRESENTATIONS -


O-2 

ENDOCRINE DISRUPTORS AFFECT REPRODUCTION 

OF THE GIANT WATER BUG: IMPLICATION FOR THE 

CONSERVATION STRATEGY 

Yoshiki Nagaba 1 , Roger Huybrechts 1 , Muhammad Tufail 2 , Hideyuki Inui 2 , 

Makio Takeda 3 

1 Biology Department, Katholieke Universiteit Leuven, Leuven, Belgium 

e-mail: roger.huybrechts@bio.kuleuven.be 

2 Organization of Advanced Science and Technology, Kobe University, Kobe, Japan 

3 Graduate School of Agricultural Science, Kobe University, Kobe, Japan, 

e-mail: mtakeda@kobe-u.ac.jp 

The giant water bug, Lethocerus deyrollei is an endangered species in Japan and 

needs conservation program. The number of bugs rapidly decreased around 1950 that 

coincided with increased pesticide use, rearrangement of farmland, and the increased 

street lights. With these ecological causes considered, we felt that more direct cause 

remained unsought. When we consider the most critical factor that regulates bug's life 

cycle, we came across possible roles of so-called endocrine disruptors. First we 

confirmed that natural insect hormones regulated the transcription of vitellogenin gene. 

Juvenile hormone is a principal endocrine factor that up regulates the synthesis 

of vitellogenin, while 20 hydroxy ecdysone inhibit the synthesis. To identify the effect 

of such chemicals, we cloned a cDNA encoding vitellogenin, the yolk protein 

precursor. We investigated the effect of four such chemicals, estradiol, bisphenol A, 

nonylphenol and octylphenol by Northern analysis. The result clearly showed that the 

former two upregulated the transcription, while the latter two down regulated at the 

common range of occurrence in the natural water systems. A number of aquatic 

Coleoptera (Dysticidae) and Hemiptera (Blastomatidae) synthesize a variety 

of steroids, mostly pregnane (Scrimshaw S and Kerfoot WC. Chemical defenses 

of freshwater organisms: beetles and bugs. In Kefoot WC, Sih A eds, Predation: 

Directand Indirect Impactson Aquatic Communities. Unive Press of New England, 

London pp240-262). This may be the reason for the rapid disappearance in the natural 

water system in Japan. The similar trend occurs now in Thailand and sooner or later 

these insects may go to extinct from everywhere unless necessary protection is 

undertaken. 

- ORAL PRESENTATIONS - 15

16 


O-3 

NEW ADVANCES IN CLONING AND EXPRESSING 

TMOF AND BTI TOXINS IN YEAST, PLANTS AND 

FUNGI 

Borovsky D. 

Florida Medical Entomology Laboratory, University of Florida-IFAS, Vero Beach, 

Florida 32962, USA, dovborovsky@gmail.com 

Advances in gene cloning and expression of peptide hormones and bacterial 

toxins were used to genetically engineer, clone and express a mosquito decapeptide 

hormone, Trypsin Modulating Oostatic Factor (TMOF; H-YDPAPPPPPP-COOH) that 

was discovered in my laboratory 20 years ago and sequenced from the ovaries 

of female Ae. aegypti. The unique property of the hormone to bind a unique gut 

receptor (recently sequence in my lab) and signal the cessation of trypsin biosynthesis 

in adult and larval mosquitoes by controlling the translation of the gene, allowed my 

laboratory in collaboration with other groups to design new cells that can control 

mosquito larvae in the field. Several unicellular cells including: chlorella and yeast 

were genetically engineered to express TMOF and efficiently controlled mosquito 

larvae in the laboratory. Because the action of TMOF is slow, it starves mosquito 

larvae, and TMOF is an excellent synergist of Bacillus thuringiensis israelensis (Bti) 

toxins we engineered yeast cells and maize pollen to expressed TMOF and Bti. Maize 

is ubiquitously used in Africa and mosquito larvae avidly eat the pollen that fall into 

the water where mosquito larvae breed. Thus, it is possible now to control mosquito 

larvae using these bio- engineering cells. These genetically engineered approaches 

will be discussed at length. 



O-4 

EFFECT OF TRYPSIN MODULATING OOSTATIC 

FACTOR (TMOF) ON TRYPSIN AND CHYMOTRYPSIN 

IN GLYPHODES PYLOALIS AND HYPHANTRIA 

CUNEA 

Ajam Hassani 1 , M., J. Jalali Sendi 1 , M. Ghadamyari 1 , D. Borovsky 2 

1 Department of Plant Protection, College of Agriculture, University of Guilan Rasht. 

Iran 

2 Florida Medical Entomology Laboratory, University of Florida, USA 

jjalali@guilan.ac.ir 

Aea-TMOF, the trypsin modulating oostatic factor is a decapeptide. It has been 

isolated from vitellogenic ovaries of the yellow fever mosquito Aedes aegypti by 

Borovsky in1985. TMOF binds to a specific gut epithelial cell receptor on the 

hemolymph side of the gut and stops trypsin biosynthesis. The hormone also affects 

larval growth, digestion of blood, eggs-maturation and oviposition rate in defferent 

insects. 

In the present study, the effect of three concentrations of TMOF (7.5, 15 and 30 G 

dissolved in water) on trypsin and chymotrypsin biosynthesis in fourth instar larvae 

of Glyphodes pyloalis and Hyphantria cunea were studied 24 h and 48 h after injecting 

TMOF. All experiments were carried out in a randome fashion with 4 treatments and 

4 replications each. Distilled water without TMOF was used as a control. Our results 

indicate that TMOF inhibits trypsin biosynthesis in H. cunea and G. pyloalis 24 h after 

the injections. In G. pyloalis lower concentrations (7.5 and 15 G) inhibited trypsin 

biosynthesis by 61.75±3.36% and 56.63±2.45%, respectively. (P< 0.05). TMOF, 

however, did not significantly affect trypsin biosynthesis at 48 h. In H. cunea on the 

other hand, at 48 h after the injection, all concentrations (7.5, 15 and 30 G per larvae) 

significantly inhibited trypsin biosynthesis by 48.2±7.1%, 43.4±6.2% and 58.4±9.5%, 

respectively (P < 0.05). No significant differences between treatments and control 

were found 24 h after injecting the hormone. Injections of TMOF did not significantly 

affect chymotrypsin in both insects. 

Because TMOF is rapidly degraded by vertebrates and it is not toxic to non target 

organisms, it could be formulated as an insecticide for the control of mosquitoes and 

lepidopteran pests. 


18 


O-5 

HOST REGULATION BY MEALYBUG PARASITOID 

AENASIUS BAMBAWALEI HAYAT (ENCYRTIDAE: 

HYMENOPTERA) 

Zain Ul Abdin 1 *, M. Ashfaq 2 , Anjum Suhail 2 , M. Jalal Arif 2 , Sohail Ahmed 2 , 

M. Ahsan Khan 2 , Dildar Gogi 2 , M. Arshad 2 , Atif Manzoor 1 , Hoor Shaina 1 , 

Erum Mubarak 1 

1. “Insect Molecular Biology Lab.” Department of Agri. Entomology, 

University of Agriculture, 38040 Faisalabad, Pakistan 

2. Department of Agri. Entomology, University of Agriculture, 38040 Faisalabad, 

Pakistan 

*Principal author to whom correspondence should be addressed: 

e-mail: zainentomology@yahoo.com, Tel: 0092419200161-170 Ext. 2906, 2904 

Successful parasitisim by the endophagous Encyrtid Aenasius bambawalei Hayat 

(Encyrtidae: Hymenoptera) has a negative impact on the fecundity of its host 

Phenacoccus solenopsis Tinsley (Pseudococcidae: Hemiptera) which ultimately leads 

to the death of the host. A significant parasitization have been observed at 27 o C±1, 

40-55 ± 5% relative humidity (RH) and 18 h light /6 h dark photoperiod which 

revealed that parasitoid took 15-20 days to complete its development after oviposition 

in the host and effective parasitization was obvious when the parasitized mealybug 

shed its wax and hardened into a leathery, brownish mummies. The study was taken 

with the aim to investigate the lethal factors responsible for the mortality of the host 

which are essential for the successful parasitism of hosts by parasitoids and it usually 

depends on gene products that the adult wasp injects at oviposition or that offspring 

produce during the course of development. These lethal factors (host regulation 

factors) usually contains venom proteins, virus-like particles, polydnaviruses, 

or ovarian proteins) present in the secretions of the ovipositing female parasitoid. 

Recent advances in Genomics and Proteomics techniques are allowing to understand 

the nature of these lethal factors/ host regulation factors of insect parasitoids and in 

this talk I will review our work specifically on venom of A.bambawalei Hayat and its 

possible use as Bioinsecticides in IPM of mealybugs. Venom of the parasitoid was 

extracted and evaluated for its Biological activity. A significant mortality of the host 

was also observed in artificially envenomated mealybugs, suggesting that venom 

of the parasitoid possess insecticidal potential. 



O-6 

CLIMATE CHANGE AND INSECT BIODIVERSITY IN 

PAKISTAN: A CRITICAL REVIEW 

Anjum Suhail, Muhammad Arshad 

Department of Agricultural Entomology, University of Agriculture Faisalabad, 

Pakistan, dranjumsuhailuaf@gmail.com 

Studies have been made to investigate the impacts of climate change on this region and 

biodiversity in Pakistan. Climate change affects biodiversity directly, and leads to the 

extinction of plants and animals that cannot keep up with rapid climate change. This is 

especially devastating in places where there are few plants or animals scattered over large 

areas. Because ecosystems are interlinked (i.e. if one plant dies out, it may affect a whole 

range of animals and insects (and possibly even other plants) that rely on it for food, shelter 

or reproduction) then even small changes may be important to the system as a whole. It is 

predicted that a major shift of flora and fauna is due to the increased temperatures and variable 

rains, especially on tropical dry forests, arid and semi-arid regions and deserts of Punjab, 

mangrove and sandy desert of Sindh, rocky desert of Balochistan and moist & dry savanna and 

pine forest of North-West Frontier Province of Pakistan. Climate change may alter the range 

and abundance of species present in an ecologic community. Nutrient cycle changes, 

community relocation, and biodiversity loss may each affect vector physiology and behavior 

(such as host-seeking characteristics and biting rates), vector populations (by increasing or 

decreasing birth or death rates), and vector migration (by changing availability of suitable 

habitats). Similarly, the ecologic factors may each directly influence pathogen dynamics for 

non-vector-borne diseases. Increase in rainfall and temperature could involve expansion of the 

malaria-carrying Anopheles mosquito into areas where malaria has never existed, or had been 

eradicated. Desertification may counter this effect, but "on the whole, increasing temperatures 

could be expected to increase the reproduction rates both of the parasites and their vectors and 

hence augment transmission of malaria. Humid and warm weather throughout July and into 

August has provided an ideal environment for bed bug, cockroaches, termite, ants to breed. It 

is expected that these insects infest thousands of Pakistani homes. 

There has been little research on Insect Biodiversity of Pakistan with reference to climate 

change. About 5,000 species of insects have been recorded (out of which 1,000 species of true 

bugs, 400 species of butterflies and moths, 256 species of grasshoppers & locusts and crickets, 

135 species of beetles & weevils, 103 species of bugs, 110 species of flies, 73 species bees, 

wasps & ants, 65 species of plant hoppers, 52 species of thrips, 49 species of termites, 48 

species of dragonflies and damselflies, 42 species of neuropterans, 32 species of cockroaches 

and few number of species in minor insect‟s orders are reported by the Department of 

Entomology, University of Agriculture, Faisalabad and University of Karachi, Pakistan, 

Biodiversity Action Plan for Pakistan, WWF - P, IUCN - P and GoP. 

Climate change impacts many of insect populations i.e., reductions in the extent of distribution 

size of some of the species that feed on pests in forest, grassland, and agricultural ecosystems. 

Pollinators‟ population in plains has been decreased and observed to shift to mountainous 

region (northern part of Pakistan) whereas bees‟ swarms which were moved from southern part 

to central and to northern part have been decreased 50 percent since last 10 years due to 

environmental change including high temperature, dust and pesticide sprays. Few years back 

in Murree, Galiat, AJK typical die back symptoms were observed in Kail pine trees during 

summer. A pine shoot beetle (Tomicus piniperda L.) was the culprit. Its population flourish 

owing to the climatic change in the area like high temperature, less snow etc. resulting a severe 

outbreak while Leaf butterfly- Kallima inachus (Nymphalidae: Lepidoptera) in Pothohar 

region has been extinct due to this climate change. 



O-7 

DIURNAL VARIATION OF AGGRESSIVE BEHAVIOUR 

IN CRICKETS 

20 

K. Faßold 1 and M.W. Lorenz 2 

1 Department of Animal Ecology I, University of Bayreuth, Bayreuth, Germany 

2 Department of Animal Ecology I, University of Bayreuth, Bayreuth, Germany, 

e-mail: matthias.lorenz@uni-bayreuth.de 

Insects exhibit distinct daily rhythms of activity. The most obvious signs 

of insect photoperiodism are the daily patterns of locomotor or feeding activity, 

although, in addition, a variety of physiological changes, including fluctuating 

hormone titres, are observed. The two-spotted cricket, Gryllus bimaculatus, is 

a nocturnal insect. In the field, flight activity and male acoustic signalling mainly 

occur in the late afternoon/early evening and the first halve of the night. Likewise, in 

the laboratory, locomotor activity of crowd-reared males and females increases during 

the late photophase and reaches a maximum during the first halve of the scotophase. 

Thereafter, locomotor activity gradually decreases, reaching low values during early 

photophase. Simultaneously to locomotor activity, the “excitability” of crickets, i.e. the 

quantity of the startle response, increases. These behavioural changes are accompanied 

by rising haemolymph lipid and, to a lower extent, carbohydrate titres, which most 

likely serve to fuel the increased physical activity. Higher titres of the energymobilising 

adipokinetic hormone (AKH) in the haemolymph of scotophase crickets 

suggest that AKH is released in higher amounts during late photo-/early scotophase 

and increases the availability of energy-rich substrates in the haemolymph, thus 

allowing the insects to “prepare” for the phase of high physical activity to come. 

Here we show that aggressive behaviour, a characteristic feature of adult male 

crickets, is also influenced by the time of day. Generally, in laboratory settings 

agonistic behaviour is measured during normal working hours, i.e. during photophase, 

where the crickets are comparatively inactive. Although male crickets almost always 

display a remarkable agonistic behaviour, an increased aggressiveness and engagement 

in fighting is observed during the scotophase. In heterologous fights, where scotophase 

males compete with photophase males, the scotophase males dominate over 

photophase males. Since haemolymph lipid titres in scotophase males are higher, there 

is a strong correlation between haemolymph lipid titre and fighting success. In 

homologous fights (photophase vs. photophase and scotophase vs. scotophase males, 

respectively), the fights between scotophase males are initiated within shorter time, are 

of higher intensity and of longer duration than the aggressive encounters between 

photophase males. In homologous fights, however, there is no clear relationship 

between haemolymph lipid titres and fighting success. On the basis of these results 

possible roles of physiological parameters (AKH-release, biogenic amines, and 

energy-rich substrates in the haemolymph) underlying the observed diurnal variations 

in behaviour are discussed. 

Supported by the Deutsche Forschungsgemeinschaft (grant No. LO 697/4-4 and 8-1) 



O-8 

STUDY ON CANNIBALISM BEHAVIOR IN LARVAL 

STAGE OF ANOPHELES STEPHENSI 

L. Tajeddin, S. Zakeri, N. D. Djadid 

Insectary of PII, Malaria and Vector Research Group (MVRG), Pasteur Institute of 

Iran (PII), Pasteur Avenue, P. O. BOX 1316943551,Tel: +98 6100955, Tehran, Iran 

We investigated the cannibalism behavior among the aquatic larval stages 

of the Anopheles stephensi (Diptera: Culicidae) with respect to food completion under 

insectary condition. The analyzed parameters were obtained as follows: 1) 

Cannibalism rate: it is the absolute number of cannibalized eggs or larvae, obtained by 

counting at the end of the experimental period; 2) Time of development: pupation and 

emergence times were measured. Both counting started at the larva hatching day and 

ended at the pupation and the emergence days respectively; 3) Adult size: Cu 1a was the 

wing vein selected to represent the adult size. The measures were taken from the wing. 

Cannibalism behavior was observed in all stages of larvae 1-4 age, being more intense 

at the end of larval development. However, in the absence of food, larvae 

of An.stephensi would have extreme tendency to cannibalism. There seems to be 

a direct correlation among the larval density per space unit and the behavior 

of cannibalism. Although it may have no straightforward application in larval control 

in field condition, but in breeding of mosquitoes in insectaries, it would be a major 

concern for having the routine breeding in progress and the required population 

density in a defined period for insecticide susceptibility tests, population genetics 

studies, and vector-parasite interaction. Possible implications of these effects for 

population dynamics of these major malaria vector mosquitoes are discussed. 


22 


O-9 

DISCOVERY OF ECDYSOGENS; THE 

BIOCHEMICALLY ACTIVATED PORPHYRIN- 

ECDYSTEROID COMPLEXES WITH POTENTIAL 

INSECTICIDE PROPERTIES 

Zhylitskaya 1 H., Litvinovskaya 1 R., Kripach 1 V., Slama 2 K. 

1 Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, 

Kuprevich str. 5/2, 220141 Minsk, Belarus; 

2 Institute of Entomology, Czech Academy of Sciences, Drnovská 507, 16100 Praha 6, 

Czech Republic zhylitskaya@gmail.com; slama@entu.cas.cz 

The polyhydroxylated, 6-keto derivatives of 7-dehydrocholesterol 

(ecdysteroids; ECD) are essential growth factors of microorganisms, animals and 

plants. In insects, they function as homeostatic factors of peripheral tissues 

synchronising gowth cycles during metamorphosis or ovarian growth cycles in the 

adults. It is generally known that ECD are effective only when injected into the 

haemolymph; previous attempts to come out with topically effective preparations 

failed due to the inability of ECD to penetrate the lipophilic layers of insect cuticle. 

We have now prepared and tested for possible topical effects several porphyrin 

derivatives, which were coupled by hydroxyboronate method with one or more 

molecules of 20-hydroxyecdysone (20-E). Unfortunately, topical assays performed 

with these porphyrin-ECD complexes in several model insect species also failed to act 

through body surface. It is more important to state, however, that the porphyrin-ECD 

complexes exhibited a hitherto unknown, delayed ecdysteroid action when injected. 

The dose-response relationships calculated for several porphyrin-ECD complexes 

revealed a range of effective concentrations which were close to that of the parental 

20-E. In larvae of the wax moth (Galleria mellonella), for example, the unwanted 

patho-physiological syndromes of "Hyperecdysonism" that are currently associated 

with higher ecdysteroid dosages, did not occur at all. By contrast, injections of 

porphyrin-ECD complexes induced a series of perfectly timed developmental events 

prerequisite for physiological course of the normal moulting process (rapid 

inactivation of larval muscles, breakdown of specifically larval tissue and organs, 

cuticle apolysis, proliferation and differentiation of pupal epidermal patterns, 

pigmentation of the pupal cuticle, resorption of exuvial fluid). In comparison with 20- 

E, the effects of porphyrin-ECD complexes were usually delayed for one or two days 

(at room temperature). We conclude, therefore, that these compounds develop a 

concentration gradient of 20-E in the body, imitating thus the naturally occurring 

endogenous ecdysteroid peaks. We are convinced that porphyrin-ECD complexes 

constitute a new group of the metabolically activated ecdysteroid complexes 

(ecdysogens), liberating the physiologically active ecdysteroid product (20-E) from the 

covalently bound porphyrin complex by intrinsic metabolic pathways. These 

properties are analogous to the previously described, biochemically activated juvenoid 

complexes (juvenogens), liberating biologically active products with juvenile hormone 

activity. Selected structural and developmental features produced by injections 

of porphyrin-ECD complexes in larvae and pupae of insects will be presented. 



O-10 

THE DESERT LOCUST CLOCK GENES: BEYOND 

CIRCADIAN FUNCTIONS 

J. Tobback 1 , B. Boerjan 2 , H.P. Vandersmissen 3 , R. Huybrechts 1 

1 Research group of Insect Physiology and Molecular Ethology 

2 Research group of Functional Genomics and Proteomics 

3 Research group of Molecular Developmental Physiology and Signal Transduction 

Department of Biology, KULeuven, Leuven, Belgium, 

e-mail: julie.tobback@bio.kuleuven.be, bart.boerjan@bio.kuleuven.be, 

hanspeter.vandersmissen@bio.kuleuven.be, roger.huybrechts@bio.kuleuven.be 

In all living organisms, behaviour, metabolism and physiology are under the 

regulation of a circadian clock. The molecular machinery (e.g. a number of oscillating 

feedback loops) has been conserved throughout the animal kingdom. Besides 

regulating the circadian timing, a variety of processes through a central oscillating 

mechanism, these circadian clock genes were found to be implicated in non-circadian 

processes such as reproduction in different insects and developmental timing in 

C. elegans. 

Recently, three circadian genes of the central feedback loops have been found 

in the desert locust Schistocerca gregaria. In the brain, period (per), timeless (tim) and 

clock (clk) transcript levels oscillate in a circadian manner. A RNA interference 

(RNAi) mediated knock down of per and tim in females resulted in a decreased 

progeny size. Currently, we are investigating the effect of a similar knock down on the 

reproductive success of males. 

More intriguing is the dose-dependent lethal effect of a clk dsRNA-mediated 

knock down in the desert locust, while clk RNAi in Drosophila melanogaster is not 

lethal and loss-of-function mutants like clk Jrk are available. Starting the RNAi in the 

fifth larval stage, delays the final moult to the adult stage, also in a dose-dependent 

manner. In adult clk RNAi treated animals, little fat tissue is present in the abdomen 

and the development of the ovaries and testes is absent or delayed. At present, the 

effect of clk RNAi on signal transduction pathways involved in reproduction and 

nutritional status are under investigation to elucidate the role of clk in Schistocerca 

gregaria. 



O-11 

THE MIDGUT BRUSH BORDER MEMBRANE IN 

INSECTS IS THE PRIMARY TARGET OF THE FUNGAL 

LECTIN (RSA) FROM RHIZOCTONIA SOLANI 

24 

Mohamad Hamshou 1,2 , Guy Smagghe 1 , Els J.M. Van Damme 2 

¹Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience 

Engineering, Ghent University, Coupure links 653, B-9000 Ghent, Belgium. 

²Laboratory of Biochemistry and Glycobiology, Department of Molecular 

Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure links 

653, B-9000 Ghent, Belgium. 

Many proteins exert toxic effects towards insects. Some of these insecticidal 

proteins belong to class of lectins, a group of proteins possessing at least one noncatalytic 

domain that binds selectively to a mono- or oligosaccharides. These 

carbohydrate-binding proteins can be found in all living organisms, ranging from 

plants to fungi, bacteria and animals. In the last decade, many plant and fungal lectins 

have been reported to have high entomotoxic effects towards insects belonging to 

different orders. 

Recently we have shown that the fungal lectin from Rhizoctonia solani (called RSA 

or Rhizoctonia solani agglutinin) has high toxic effects towards cotton leafworm 

Spodoptera littoralis (Hamshou et al. 2010) and pea aphid Acyrthosiphon pisum (on 

unpublished data). More detailed studies have now been performed to understand the 

mode of action of RSA toxicity. 

First, the binding and the interaction of the lectin with A. pisum and S. littoralis tissues 

have been examined using cross sections of A. pisum nymphs and S. littoralis larvae 

fed on an artificial diet containing FITC-labeled RSA. Analysis of the section using 

fluorescence microscopy revealed that RSA is bound to the epithelial brush border of 

the midgut in both insects. 

Second, the cellular toxicity and interaction of RSA with different insect cell lines 

were investigated. RSA showed a high toxicity towards all cell lines used in this study 

(CF-203 midgut cells, S2 embryonal cells, Bm5 ovarian cells and CPB fat body cells). 

The lectin effect on cells was dose-dependent and about 50% toxicity was observed for 

0.3 µM RSA. Analyses using the confocal fluorescence microscope demonstrated that 

FITC-labeled RSA was not internalized in any of these cell lines, but was bound to the 

cell surface which nicely correlates with the fact that RSA bound to the surface of the 

epithelium cells of the midgut in pea aphid nymphs and cotton leafworm larvae. 

Therefore we focused our research on the CF-203 cell line which is derived from 

midgut cells. The binding of different lectins with different carbohydrate-binding 

specificities to CF-203 midgut cells was compared. Interestingly, cell death of CF203 

cells caused by RSA was accompanied with DNA fragmentation and the effect was 

caspase-7, -8 and -9 dependent but caspase-3 independent. 

More studies are necessary to understand the exact mechanism behind the toxicity 

of this lectin and to investigate the possibility of using RSA for bioengineered insect 

resistance in important agronomical crops. 

Hamshou, M., Smagghe, G., Van Damme, E.J.M. (2010). Entomotoxic effects of fungal lectin from 

Rhizoctonia solani towards Spodoptera littoralis. Fungal Biology 114, 34-40. 



O-12 

COMMUNITY ECOLOGY AND PEST CONTROL 

POTENTIAL BY CARABID BEETLES IN 

AGRICULTURAL LANDSCAPES 

Stefaniya Kamenova, Bertrand Gauffre, Vincent Bretagnolle, Pierre Taberlet, Eric 

Coissac, Manuel Plantegenest 

Effective crop protection with minimal impact on the environment is now 

a major concern. Promoting biological control of pests by natural enemies has often 

been proposed as a possible alternative strategy to ensure crop protection. Hence, 

research need now to identify situations favourable to biological control. It is therefore 

essential to understand the factors structuring the community of predatory insects. 

Carabid beetles are usually designated as natural control agents in agricultural areas 

but the lack of precise information in both on their diets and on factors structuring their 

community, limits our ability to draw clear conclusions. To search for factors 

determining the presence and regulation potential of carabid beetles, it is important 

to focus on trophic relationships across the community. Until then, such an approach 

has been difficult to achieve because of methodological limitations. The recently 

developed molecular technologies such as barcoding and throughput sequencing 

provide exciting opportunities for large-scale studies of trophic relationships. 

Combining these new molecular tools with a more traditional isotopic analysis, we 

investigated the diet and the trophic niche of a community of the seven most abundant 

carabid species in a french agricultural landscape. Individuals were collected between 

April and June, in 12 wheat and oilseed rape fields. Isotopic signatures of specimens 

were obtained by a mass spectometer analysis of 13C and 15N ratios. In parallel DNA 

from faeces of the same indviduals was amplified with three different sets of primers 

and a set of blocking primers was used to prevent the amplification of predator DNA. 

One set of primers, more specific to the Carabidae familly, was developed to detect the 

possible intra-guild predation. Pooled amplicons from each individual were sequenced 

with the Solexa technology (Illumina). 

This is the very first case such a complementary approach is applied on the community 

of carabid beetles. The quantity of generated data opens a vast field of research and 

many new prospects for the development of large-scale management strategies. 


26 


O-13 

ECO-PHYSIOLOGICAL CHALLENGES AND 

COMPATIBILITIES AMONG DESERT INSECTS 

Meera Srivastava, M.M. Saxena 

P.G. Department of Zoology, Govt. Dungar College, Bikaner -3340 01, Rajasthan, 

India, meerasrivastava@rediffmail.com 

In animal kingdom arthropods constitute the biggest group among which 

insects alone share the major chunk. Insects are the most versatile class of animals that 

inhabit all concern able environments and occupy any kind of possible ecological 

niche. In food habits they range from detritivores, herbivores, scavengers, to parasites 

and in habitat they are found in soil, on the soil, in water, in air, on high altitudes and 

frigid zones to warm tropics, in orchids to rancid mud. 

A hot desert offers conditions that are not tenable to many organisms, particularly cold 

blooded animals. Despite, variety of insects are found in such a harsh environment, 

and authors have documented over hundred insect species of land aquatic insects from 

the Indian desert. The land insects are found to withstand extremes of temperature, 

dryness, and general dearth of food while aquatic one face periodic dryness of water 

bodies and deficiency of oxygen. To meet such challenges such insects are found to be 

highly adapted through physiology, anatomy, ecology and behaviour. 



O-14 

POTENTIAL OF HERBAL MEDICINE PRODUCTS FOR 

PREVENTING FRUIT INFESTATION BY CODLING 

MOTH NEONATES 

Maciej A. Pszczolkowski 1 , Kevin Durden 1 , Samantha Sellars 1 , Brian Cowell 1 , 

John J. Brown 2 

1 William Darr School of Agriculture, Missouri State University, 

Mountain Grove, MO 6571, USA, e-mail: MPszczolkowski@missouristate.edu 

2 Department of Entomology, Washington State University, Pullman, 

WA 99164, USA 

Effects of popular herbal medicine products (dietary supplements) on fruit 

infesting behavior of neonate codling moth, Cydia pomonella, larvae were studied. 

The supplements based on Ginkgo biloba, Artemisia sp., and Tanacetum sp. showed 

the most promising potential for preventing fruit infestation. Five secondary 

metabolites from these plants were identified as insect feeding deterrents. 


28 


O-15 

BIOLOGICAL PARAMETERS AND PREFERENTIAL 

FEEDING RESPONSE OF COCCINELLA 

UNDECIMPUNCTATA L. ON THREE APHID SPECIES 

J.M.Mari 

Department of Plant Protection, Sindh Agriculture University Tando Jam Pakistan 

janmarreedr@yahoo.com and janmarree@gmail.com 

Biological Parameters and Preferential feeding response of Coccinella 

undecimpunctata L. on three aphid species was observed in the department 

of Entomology, Sindh Agriculture University, Tando Jam Pakistan. The data indicated 

that the mean incubation period of 11-spotted eggs laid by females reared on maize, 

alfalfa, and mustard aphids were 6.50, 8.20, and 7.00 days respectively. For fecundity 

mean number of eggs per female was 621, 616, and 601 when reared on maize, alfalfa, 

and mustard aphids, respectively. The hatch percent of eggs was 70.21, 68.11, and 

66.03 when reared on safflower, alfalfa, and mustard aphids. The duration values 

in days for first, second, third and fourth instars ranged between 4.50, 5.60, 7.90 and 

8.01, respectively on maize aphid. For alfalfa aphid it was 4.90, 5.25, 6.10 and 7.80, 

respectively and for Mustard aphid the values were 3.99, 5.23, 5.66 and 8.50, 

respectively. Pupal period in this study ranged between 4.1, 4.59 and 4.30, respectively 

for maize, alfalfa, and mustard aphids. Pre-oviposition and post-oviposition periods 

of 11-Spotted beetle in the study on maize aphid were 4.02 and 24.22, respectively. 

For alfalfa aphid it was 3.51 and 3.91days, respectively on mustard aphid it lasted 3.91 

and 23.62 days respectively. Adult (female) life span of the beetle under present study 

was 35.00, 50.00 and 40.90 days for maize, alfalfa and mustard aphid, respectively, for 

adult (male) it was 30.00, 42.00 and 32.00, respectively. Preferential feeding response 

of beetle revealed variation in their voracity. For instance, 1 st , 2 nd , 3 rd and 4 th instar 

larvae were 5.50, 20, 30 and 41.25 for maize aphid, respectively. On the other hand, 

for alfalfa aphid it was 10.95, 36, 46.5 and 57, respectively and for mustard aphid it 

was 6.55, 25.5, 36.5 and 46¸ respectively. Preferential feeding response for aphid 

feeding rate per day for adult (female and male) was 24.62 and 24.26 for maize aphid, 

respectively. It for alfalfa aphid was 30.08 and 26.19, respectively and for mustard 

aphid it was 20.72 and 24.18¸ respectively. Coccinella undecimpunctata grubs and 

adults were voracious feeders on safflower, alfalfa, and mustard aphids. This beetle 

has great potential for use biologically control of these three aphid species. Further 

studies on predator efficiency of Coccinella undecimpunctata may be conducted with 

other important aphid pests such as cotton, berseem, corn leaf, chickpea aphids, etc. 

Keywords: Biology, feeding response, aphid and 11-spotted 



O-16 

CAPILLARY ELECTROPHORESIS IN SCREENING 

OF BIOACTIVE COMPOUNDS - PHYSICOCHEMICAL 

PROPERTIES AND MOLECULAR INTERACTIONS 

Alicja Kluczyk 

Faculty of Chemistry, Wroclaw University, Wroclaw, Poland, kluczyk@wchuwr.pl 

Interactions between biomolecules constitute the most important, as well as the 

most interesting part of mechanisms of life at molecular level. There are several 

methods of identification of interacting components, including two-hybrid system, 

application of reactive linkers, chromatography and mass spectrometry. The selection 

of the method depends on the information about the investigated biological system, its 

level of complexity and availability of the interacting components. Best results are 

usually obtained for methods combining separation and identification of substances. 

Capillary electrophoresis (CE), method based on separation of ionized molecules in 

narrow-bore capillary according to their mobility in electric field, could be used to 

investigate the interactions between molecules [1]. The method allows the use of 

buffer solutions in the whole range of pH, as well as several additives, including 

inorganic salts, organic solvents and surfactants. Several strategies have been 

developed and successfully applied to analyze the affinity of molecules. Depending on 

the method, either a new signal is observed in electrpherogram, or the mobility of the 

analyte changes, or a vacancy appears in the region where a component was consumed 

to form a complex. CE could be used not only to identify the interactions, but also to 

measure the binding constants [2]. The advantages of CE include minimal 

consumption of reagents (comparable to the amount required for mass spectrometry 

analysis) and broad range of experimental conditions which could be used to 

determine the influence of environmental conditions on the interactions. 

The main areas of application of CE include interactions of drug molecules with 

proteins, immunoaffinity studies, chiral differentiation, incorporation of chemicals into 

model membranes and complexing abilities towards metal ions [3-6]. The general 

strategies and practical applications will be discussed to present the method and its 

advantages. 

References 

[1] Galbusera Ch. et al. (2003) Curr. Opin. Biotech., 14, 126. 

[2] Tanaka Y. et al. (2002) J. Chromatogr. B, 768, 81. 

[3] Ostergaard J. et al. (2003) Electrophoresis, 24, 2903. 

[4] Gawron A. et al. (2000) Electrophoresis, 21, 2067. 

[5] Corradini D. et al. (2004) Chromatographia, 60, S125. 

[6] Miki S. et al. (2005) J. Chromatogr. A, 1066, 197. 



O-17 

RE-EXAMINATION OF THE FACTORS CONTROLLING 

BODY-COLOR POLYPHENISM IN DESERT LOCUSTS 

30 

Seiji Tanaka 

1 Locust Research Laboratory, National Institute of Agro-biological Sciences at 

Ohwashi (NIASO), Tsukuba, Ibaraki 305-8634, Japan 

Many studies have been carried out to investigate various aspects of phase 

variation and body-color polyphenism in locusts and grasshoppers (Uvarov, 1966), and 

much information is available especially for the desert locust, Schistocerca gregaria. 

Juvenile hormone induces solitary green body color and a neuropeptide, [His 7 ]corazonin, 

is involved in the control of dark colors and other phase-dependent 

morphological traits (Tanaka, 2006). It has been widely accepted that humidity is a 

primary factor controlling green-brown polyphenism in this locust under solitary 

conditions (Pener and Simpson, 2009). Solitary nymphs are green at high humidity, 

whereas those at low humidity assume various brown (non-green) body colors. During 

a field trip to Mauritania, however, we encountered green solitary nymphs of this 

locust even under considerably dry conditions (Tanaka et al., 2010). Crowding is 

another important factor inducing black patterns characteristic of gregarious forms. 

Black patterning is often assumed to be a gregarious (and transient) trait that can be 

induced by tactile, olfactory and visual stimuli from other individuals in the laboratory 

(Lester et al., 2005). In the field, however, some solitary nymphs appear to develop 

black patterns and such individuals look fairly inconspicuous on brown vegetation 

(Tanaka et al. 2010). I re-investigated how body-color polyphenism of this locust was 

controlled in the laboratory, and found that 1) neither humidity nor food quality 

influences green-brown polyphenism, 2) this locust exhibits homochromic responses 

to the background color, 3) black patterns appear even among solitary (isolated) 

hoppers under certain conditions and 4) visual stimuli instead of olfactory stimuli play 

an important role in inducing gregarious body coloration. This paper presents 

experimental results documenting these findings and discusses the significance 

of body-color polyphenism in this locust. 

References 

Lester R.L. et al. (2005) J. Insect Physiol. 51,737. 

Pener M.P., Simpson, S. (2009) Adv. Insect Physiol. 36, 1. 

Tanaka S, (2006) Appl. Entomol. Zool. 41, 179. 

Tanaka S. et al. (2010) Appl. Entomol. Zool. 45, 643. 

Uvarov, B. (1966) Grasshoppers and Locusts Vol. I. Cambridge Univ. Press. 



O-18 

BIORATIONAL CONTROL OF ARTHROPOD PESTS 

WITH EMPHASIS ON THE USE OF THE CHITIN 

SYNTHESIS NOVALURON 

Ishaaya I., Lebedev G., A., Horowitz, A. R. 

Department of Entomology, Agricultural Research Organization, The Volcani Center, 

Bet Dagan 50250, Israel 

E-mail: vpisha@volcani.agri.gov.il 

Efforts have been made during the past three decades to develop insecticides 

with selective properties that act specifically on biochemical sites present in 

a particular insect group, but whose properties differ from those present in the 

mammalian system. This approach has led to the discovery of compounds that affect 

the hormonal regulative of molting and developmental processes in insects such 

as ecdysone agonists, juvenile hormone mimics and chitin synthesis inhibitors. 

The search for potent acylureas has led to the development of novaluron (Rimon) 

developed by Makhteshim Chemical Works. The LC-50 value of novaluron on 3 rd - 

instar Spodoptera littoralis fed on treated leaves is approximately 0.1 mg a.i./liter. This 

value resembles that of chlorfluazuron and is tenfold lower than that of teflubenzuron. 

Novaluron affects nymphs of Bemisia tabaci to a much greater extent than does 

chlorfluazuron and teflubenzuron. Artificial rain, at a rate of 40 mm/h, applied 5 and 

24 h after treatment in a cotton field had no appreciable effect on the potency 

of novaluron on S. littoralis larvae or B. tabaci nymphs. Hence, novaluron can be used 

in tropical areas and in rainy seasons. In general, benzoylphenyl ureas had no direct 

effect on parasitoids and phytoseiids and are considered to have a mild effect on other 

natural enemies. Novaluron has no cross-resistance with conventional insecticides, 

the JH mimic pyriproxyfen and neonicotinoids. As such, it is considered an important 

addition in pest management programs. 


32 


O-19 

LETHAL AND SUBLETHAL EFFECTS OF THREE 

INSECT GROWTH REGULATORS, PYRIPROXYFEN, 

CYROMAZINE AND METHOPRENE ON THE 

BOLLWORM, HELICOVERPA ARMIGERA 

Farshbaf Pourabad, R. 1 , Askari, G. 2 , Ebrahimi, L. 3 

1 Department of Plant Protection, University of Tabriz, Tabriz, Iran, 

rfpourabad@yahoo.com 


askarigh@gmail.com 


ebrahimi.laleh@gmail.com 

Bollworm, Helicoverpa armigera, is one of the most important pests of crops 

particularly cotton in Iran. Lethal and sublethal effects of three IGR insecticides, 

pyriproxyfen, methopren and cyromazine, were studied on H. armigera in laboratory 

condition. Bioassays were carried out on second instars. Larvae were fed 

by insecticide-treated artificial diet for five days. Experimental concentrations for 

pyriproxyfen and methopren treatment were 2000, 4000 and 8000 ppm and for 

cyromazine treatment were 500, 2000, 4000 and 8000 ppm. Lethal and sublethal 

effects were recorded daily until 22 days. Mortality of the treated insects was higher 

than 90% for all compounds in 4000 and 8000 ppm. In 2000 ppm mortality 

percentages were 77.25, 82.5 and 80% for methoprene, pyriproxyfen and cyromazine, 

respectively. Insect mortality was 80% in concentration of 500 ppm for cyromazin 

which demonstrated high lethal effect of this insecticide against H. armigera, 

in comparison with two other compounds. Sublethal effects of the insecticides 

included deformation of the larvae and pupae, increase of development duration 

especially by pyriproxyfen treatments, blister-like lesions by cyromazine treatments 

in third and fourth larvae and presence of abdominal legs in pupae by pyriproxyfen 

treatments. In pyriproxyfen and methopren intermediate stages, larva-pupa and pupa – 

adult were formed. In these treatments adult with wrinkle wings were formed. Results 

of this study revealed that pyriproxyfen, cyromazine and methoprene were lethal 

against H. armigera in life duration, especially in cyromazine treatment which caused 

60% death of larvae in third day of experiment. 



O-20 

BIODIVERSITY OF CHIRONOMIDAE (DIPTERA) AND 

GENOME RESPONSE TO STRESS AGENTS IN THE 

ENVIRONMENT 

Michailova P. 1 , Szarek-Gwiazda E. 2 , Kownacki A. 2 Warchałowska-Śliwa E. 3 

1 Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 1 

Tzar Osvoboditel boulv., Sofia, Bulgaria 

2 Institute of Nature Conservation, Polish Academy of Sciences, al. Mickiewicza 33, 

Krakow, Poland, e-mail: szarek@iop.krakow.pl 

3 Institute of Systematic and Evolution of Animals, Polish Academy of Sciences, 

Sławkowska 17, Krakow, Poland 

The effects of heavy metals on the biodiversity and genome level 

of Chironomidae larvae (Diptera, Insecta) were evaluated in the strongly contaminated 

water environments. No influence on the Chironomid species diversity was found. 

However, the higher concentrations of trace metals (Cd, Cu, Pb, Mn, Cr, Fe and Ni) 

affect the genome of 5 Chironomidae species: Chironomus bernensis, C. plumosus, 

C. sp.1., Kiefferulus tendipediformis (cytotype 2), Glyptotendipes gripekoveni. 

Genome instability of Chironomid larvae was manifested by two ways: 1. Fixed 

chromosome rearrangements; homozygous inversions and tandem fusions create new 

gene linkage groups and show an intensive microevolutionary process of species. 2. 

Somatic structural and functional alterations in salivary gland chromosomes of all 

studied Chironomidae species: inversions, deficiencies, deletions, breaks; decreasing 

the functional activity of cell key structures: Balbiani rings (BRs) and Nucleolar 

Organizer (NOR). Species – specific response was found: new puffs, decondenzation 

of centromere heterochromatin, asynapsis, “dark knob” at the telomeres of the 

chromosome and etc. Detecting somatic rearrangements in salivary gland 

chromosomes of these widely distributed aquatic insects is potentially one of the best 

validated bioassay and can be used as a cost effective early warning signals 

of environmental damage in freshwater basins. 

Acknowledgement: project D0 – 02 -259/08 and exchange program between BAS and PAS. 


34 


O-21 

SUSCEPTIBILITY OF DIFFERENT VARIETIES OF 

COTTON TO INFESTATION WITH SOME SUCKING 

INSECTS UNDER FIELD CONDITIONS IN EGYPT 

M. I. Mogahed 

Pests & Plant Protection Dept. 

National Research Centre. Tahrir St., Dokki, Cairo ,Egypt 

E-mail: mogahedprof2007@hotmail.com 

A field experiments were conducted to study the susceptibility of eight cotton 

varieties( Aleppo 33; McNair 308; Tashkent 3; McNair 307; McNair 235; TamCot 

CAMD; McNair 220 As imported var. and Moubarak 93 as local var.) to infestation 

with cotton aphids and cotton whitefly.The results showed that the aphid population 

had three main peaks (1st peak in July, 2 nd peak in the 2 nd week of August and the 

minor peak (3 rd ) occurred at the last week of August). Aphid population showed high 

reduction with low average of infestation (17.9-24.0%) at May and June with Temp. 

(22.9-25 o C), R. H. (48-56%) Wind speed (1.8-2.0m/sec.) and sunshine duration (13.6- 

14.1 hr and mid of September of cultural season. 

The most attractive varieties of cotton to A. gossypii were Var. Aleppo 33, var. 

McNair235 and var. TashKent 3 followed by var. McNair 307, whereas the least 

attractive varieties of cotton were var. Tamcot CAMD and var. McNair 220, 

comparing with the control (var. Mubarak 93).There were high significant difference 

between infestation of cotton varieties with A.gossypii . 

The population of cotton whitefly, B. tabaci had three main peaks in growing 

season (July, August and September) with Temp.(26.2-26.6 o C), R. H. (68.3- 

69.0%),wind speed (0.77-0.90 m/sec) and sunshine duration (12.4-13.9 hr), 

however,the lowest population of whitefly was recorded in May with Temp. 23.0 o C), 

R.h. (51.7%), W. speed (1.7 m/sec.) and sunshine duration (13.6 hr ). Both A. gossypii 

and B. tabaci reached peak population in the third and fourth week of August 

(2008).The local variety (Moubarak 93) was the most tolerant against infestation with 

A. gossypii and B. tabaci..It was found that the rates of cotton yield were decreased 

with increasing infestation rates of aphid or white fly. 

Keywords: Susceptibility, Cotton varieties, Sucking insects, Environmental factors, Yield 



O-22 

THE EFFECT OF SOME MICROBIAL CONTROL 

AGENT AGAINST TOMATO PESTS U IN EGYPT 

Magda Sabbour 

Pest and Plant Protection Department, National Research Center, El-Tahrir Street 

(El Behoose St., P.O. Box 12622), Dokki, Cairo,Egypt. 

e-mail: sabbourm9@yahoo.com 

Two microbial control agents Nomuraea rileyi(N.r) and Paecilomyces 

fumosoroseus (P.f) were evaluated against Bemisia tabaci and Aphis gossypii 

in tomato under laboratory, greenhouse and field conditions. Results showed that 

percentages of infestation decreased after treatment with the fungi. Under laboratory 

conditions, LC50 values for B. tabaci were 125.7 x10 8 spores/ml for (N.r) and 137.4 

x10 8 spores/ml for (P.f) LC50, values for A. gosspii were 129.1 x10 8 spores/ ml for 

(N.r) and 139.8 x10 8 spores/ ml for(P.f). In the greenhouse LC50 values for B. tabaci 

were 56.4 x10 8 spores/ml for(N.r). and 76.6x10 8 spores/ml for(P.f)., and for 

A. gossypii were 68x10 8 spores/ml for(N.r). and 88x10 8 spores/ml for (P.f). In field 

conditions the percentage of infested plants with B. tabaci and A. gossypii significantly 

decreased after treatments with both(N.r). and(P.f). as compared with the control 

treatment in El-Esraa farm (Nobaria region) and in El-Kassaseen (Ismailia). Weight of 

tomato yield in El-Esraa was 3487 and 3448 kg/ faddan when (P.f) was applied as in 

comparison with 1000 kg/ faddan in the control and in El-Kassaseen 3531 kg/feddan 

when (N.r) was applied and 3400 kg/feddan kg/feddan when(P.f). was applied as 

compared to 977 kg/ feddan in the control. 

Keywords: Nomuraea rileyi Paecilomyces fumosoroseus, Bemisia tabaci, Aphis gossypii, microbial 

control. 



O-23 

SCREENING OF INDUCIBLE IMMUNE PROTEINS FROM 

SILKWORM BOMBYX MORI BY TWO DIMENSIONAL 

ELECTROPHORESIS 

A. Mallikarjun Gadwala 1 , B. V Shyam Kumar 2 , C. Neetha Kari 3 , D. Narayan Moger 4 , 

E. B.B.Kaliwal 5 

1 Dept of Sericulture, karnatak university, Dharwad. INDIA, email:gspmalli@gmail.com 

2 Dept of Biotechnology and Microbiology, karnatak university, Dharwad. 

e-mail:vootlashyam@gmail.com 

3 Department of Biotechnology, University of Agricultural sciences, Dharwad, INDIA , 

Amongst all the living species insects occupy more than 50% of the current 

biodiversity. Insects are evolutionarily successful organisms and occupy almost all 

habitats in nature. Insect immune response is comprised of cellular and humoral 

components; the humoral component comprises of antibacterial proteins which are 

inducible. Antibacterial proteins are an important component of insect immunity. 

Among the insects, only the dipterans (Drosophila and various mosquito species) have 

been widely investigated for their immune responses towards diverse pathogens. In the 

present study, we have attempted to investigate the immune transcriptome of the 

lepidopteron insect silkworm Bombyx mori L. We studied the bacterial infected 

silkworm haemolymph by SDS-PAGE & 2-DE (Two- Dimensional Electrophoresis) 

to identify the immune proteins. Differentially expressed protein were isolated and 

characterized by MALDI-TOF analysis. A novel antibacterial protein in 5 th instar 4 th 

day of immunized haemolymph of 31.8 KDa M.W. was characterized and concluded 

as an active protein involving immunity in B.mori. 

36 



O-24 

LEPIDIOTA MANSUETA BURMEISTER (COLEOPTERA: 

SCARABAEIDAE): A NEW SEVERE KEY PEST IN 

MAJULI ISLAND, ASSAM 

Bhattacharyya Badal 1 , Bhuyan Utpal 2 , Baruah A.A.L.H. 3 , Pujari D. 4 , Saud R.K 5 

All India Network Project on White grubs and other soil arthropods 

Department of Entomology, Assam Agricultural University, Jorhat-785013, Assam, 

India, Phone: +91-9435648957, Fax: + 91- 376-2310037, 

email: badalassam@gmail.com 

Field and laboratory investigations were conducted at Assam Agricultural 

University, Jorhat, Assam during 2005-2010 on different aspects of Lepidiota 

mansueta Burmeister (Coleoptera: Scarabaeidae). Of late, this species has appeared 

as a severe key pest of many field crops and its incidence was recorded in fifteen 

villages in Majuli, the largest mid-river deltaic island of the world located in Jorhat 

district of Assam, India. While estimating the damage potential of this pest, it was 

observed that the third instar larvae were found to cause the heavy damage to the 

crops. The most severely affected crops were potato, sugarcane, Colocasia and green 

gram and the extent of damage was found to vary from 42-48, 15-20 , 35-40 and 30-35 

per cent respectively. L. mansueta was first detected in 2008 with high flight activity 

(flight year) and subsequently in 2010. There was no adult emergence during 2009 and 

only the larvae were found under the soil, indicating the biennial life cycle of the pest. 

The duration of egg, grub and pupal stages were found to vary from 12-17, 635-671 

and 28-35 days respectively in laboratory condition. There were three larval instars. 

Duration of first and second instar grubs were found to be relatively short and ranged 

from 35-45 and 55-63 days respectively. The third instar grubs showed a prolonged 

developmental period which varied from 545 to 563 days. The adults were found to be 

phototactic and maximum flight activity of the beetles was observed during 18.30 

to 20.30 hours. Adults of both sexes of this insect were not observed feeding on any 

plants in the field unlike other white grub species, probably due to the presence 

of degenerated mouth parts (atrophied).The host specificity test, gut physiology and 

mouth parts dissection further confirmed the nonfeeding behaviour of the adults and 

on the basis of these findings, L. mansueta has got the unique distinction of first Indian 

phytophagous white grub species with nonfeeding adults. Severity of this insect 

in Majuli island may be attributed to the presence of abundant thatch zone of that 

locality with high organic carbon content of the soil. Among the weeds, Elusine indica 

was found to be the main host of the larvae of the pest. The recent upsurge of this pest 

in different crops is probably due to cultivation in the fallow low-lying areas which 

were the original inhabiting site of the pest. The possible effect of climatic change 

on the activities of migratory predatory birds of L. mansueta has a strong relevance 

on the recent outbreak of this pest in Majuli Island. 


38 


O-25 

TOXICITY OF FIVE MEDICINAL PLANT OILS TO 

WOOLLY APPLE APHID, ERIOSOMA LANIGERUM 

(HOMOPTERA: APHIDIDAE) 

M. Ateyyat 1 , M. Abdel-Wali 2 , T. Al-Antary 3 

1 Department of Plant Production and Protection, Faculty of Agricultural Technology, 

Al-Balqa’ Applied University, Al-Salt 19117, Jordan, e-mail: ateyyat@bau.edu.jo 

2 National Center of Agricultural Research and Transfer, Al-Baqa’, Jordan 

3 Plant Protection Department, Faculty of Agriculture, University of Jordan, Jordan 

The toxicity of oil extracts of five medicinal plants was evaluated against 

the woolly apple aphid, Eriosoma lanigerum, (Homoptera: Aphididae). Oils were 

extracted from Achillea santolina, Artemisia seiberi, Salvia officinalis, Thymus 

Vulgaris and Senecio vernalis. Each oil extract was used at three concentrations, 100 

ppm, 1000 ppm and 100,000 ppm. Mortalities were corrected using Abbott‟s formula 

and TableCurve fitting program(Jandel Scientific ® ) was used to predict the LC50 

of each extract. Results revealed that LC50 after 24 hr post treatment is 7011, 6161, 

6667, 9090 and 9034 ppm for A. santolina, A. seiberi, S. officinalis, T. Vulgaris and 

S. vernalis, respectively. As time response, LC50 was obtained at


O-26 

THE PURPOSE OF PUDDLING OF MALE PAPILIO 

BUTTERFLIES 

Asaoka K., Hata T., Ito T., Hagiya H., Niihara K., Yokohari F. Inoue T.A.* 

*Shin-Machi 3-24-14, Kokubunji, Tokyo, 185-0004, Japan 

*e-mail: inoueatp@affrc.go.jp 

We examined the purpose of puddling behaveior of male Japanese Papilio 

butterflies from the aspects of morphology, behavior, electrophysiology and 

measurement of the fluctuation of the metal ion concentrations between the water 

given to the butterflies and urine of butterflies. 

By morphology, we already found taste sensilla from inside of proboscis of butterflies, 

additionally, we could find the structures that were considered olfaction sensilla on the 

antenna of butterflies. 

By behavioral research in the field, male Papilio butterflies selected Na ion taste 

among Na, K, Ca and Mg, most preferred concentration of Na ion was 10mM, and 

anion were not affected to this preference. And these butterflies attracted to the smell 

of H3N and H2O, in contrast, H2S and AceticAcid did not attract butterflies. 

By electrophysiology to the taste sensilla in proboscis using tip-recording, these 

sensilla evoked clear spike trains to Na ion solutions and did not evoked to K, Ca and 

Mg. To the olfact-sensilla-like structures using single-cell recording, these evoked 

clear spike trains to H3N and H2O, and not H2S and AceticAcid. 

By measurement of the fluctuation of the metal ion concentrations between the water 

given to the butterflies and urine of butterflies, we found that all butterflies who were 

given 13mM-Na ion water ingest Na ion, in contrast, some butterflies who were given 

more Na ion concentration water excrete Na ion. Na ion concentration in field water 

was up to 6mM, so to much Na ion concentration water must be harmful for 

butterflies. 

As conclusion, male Japanese butterflies puddle to try to get Na ion from the natural 

water, and to search the good places for puddling, they seemed to use the smell of H3N 

and H2O. 



O-27 

EFFECTIVENESS OF TRANSGENIC BT COTTON WITH 

CRY1AC AGAINST SPOTTED BOLLWORM EARIAS 

INSULANA (BOISD.) AND PINK BOLLWORM 

PECTINOPHORA GOSSYPIELLA (SAUND.) 

40 

Muhammad Arshad, Anjum Suhail, Zain ul Abdin, H. Fahim udin Shah 

Department of Agricultural Entomology, University of Agriculture Faisalabad, 

Pakistan, drarshaduaf@uaf.edu.pk 

One option to reduce the insecticide use on cotton is the exploitation 

of transgenic Bt cotton as a component of integrated pest management. Bt cotton, the 

first transgenic non-food crop, has provided a specific, safe and effective tool for the 

control of lepidopteran pests. Transgenic Bt cotton containing Cry1Ac toxic gene from 

Bacillus thuringiensis was evaluated for resistance against spotted bollworm Earias 

insulana (Boisd.) and pink bollworm Pectinophora gossypiella (Saund.) in Punjab 

province, Pakistan during two cotton seasons. We found no significant differences 

in spotted bollworm egg density between Bt and conventional cotton but larval density 

was significantly lower in unsprayed Bt cotton than unsprayed non-Bt cotton plots and 

insecticides were applied in sprayed non-Bt plots to keep the pest population below the 

threshold level. The rosette flowers density caused by pink bollworm infestation and 

number of pink bollworm larvae found in bolls of Bt cotton were significantly lower 

than non-Bt cotton plots. We conclude that transgenic Bt cotton is more effective 

in season-long control of spotted and pink bollworm. 



O-28 

INVESTIGATION RESISTANCE OF POPLAR SPECIES 

AND CLONES TO MONOSTEIRA UNICOSTATA (HET.: 

TINGIDAE) IN KARAJ OF IRAN 

Babmorad, M. 1 ; Sadeghi; S.E. 1 ; Hesamzadeh, M. 1 , Satar, Z. 

1-Research Institute of Forests and Rangelands, Iran 

babmorad@rifr-ac.ir 

Poplar lace bug, Monosteira unicostata (Het.: Tingidae) is one of the most 

important insect pest of Salicaceae family especially poplars in nurseries and 

plantations of Iran. During 2003-2005, damage of poplar lace bug and also number 

of eggs laid of the pest were studied on 15 native & exotic poplar clones belonging to 

Populus nigra L, Populus deltoides Marsh, P. x. euramericana (Dode) Guinier and 

also Populus alba L. in Karaj. area. In the first step, clones were planted as 

randomized complete block design with 3 replications In late March of 2002. Every 

experimental plot included 16 seedlings. In each plot, 4 seedlings (included 4 leaves of 

each seedling) were randomly evaluated for number of female adults ovipositing at 30 

day intervals As well as for lace bug damage at 15 day intervals, starting in early June 

until early October. The data modified and were analyzed by SAS software and 

Duncan test (α=1%) was applied to compare the means. Based on the three years 

results, there was significant difference (α=1%) between poplar species and clones. 

Of all poplar clones, P. alba 44.9 and P. alba 58.57 had the highest susceptibility 

against poplar lace bug.and P. deltoides 77.51, P. deltoides 73.51, P. deltoides 69.55 

and also P. x. euramericana 561.41 showed the highest resistance to poplar lace bug 

compared with other tested clones. 

Keywords: poplar (populus), species, clones Poplar lace bug, Monosteira unicostata, egg, damage, 

Karaj, Iran 


42 


O-29 

INSECT NEUROPEPTIDE LEUCOPYROKININ 

STIMULATES INTERFERON PRODUCTUIN IN RATS 

Plech Andrzej 1,2,3 , Konopińska Danuta 4 , Rykaczewska-Czerwińska Monika 1 

1Department of Pharmacology, Medical Faculty in Zabrze of the Medical University of 

Silesia in Katowice, 28 Jordana Str., 41-808 Zabrze, Poland; 

2 Cardinal August Hlond Upper Silesian College of Pedagogical Education, Faculty of 

Public Health, 19 Powstańców Str., 41-400 Mysłowice, Poland; 

3 Silesian College of Economics and Administration, Faculty of Physiotherapy, 12 

A.Frycza – Modrzewskiego St., 41-907 Bytom, Poland, 

4 Faculty of Chemistry, Wrocław University, 14 F Joliot-Curie Str., 

50-383 Wrocław, Poland 

Insect-derived peptides: octapeptide leucopyrokinin (LPK) and its analog [2-8]leucopyrokinin 

([2-8]-LPK) exert several biological effects in mammals (Plech A. et 

al., 1997, Pol. J. Pharmacol., 49, 119-126), (Rykaczewska-Czerwińska M. et al., 2001, 

Comp. Biochem. Physiol. Part C, 130, 271-279). These peptides applied directly into 

the lateral brain ventricle (icv) induced evident antinociceptive effect in rats (Plech A. 

et al., 1997, Pol. J. Pharmacol., 49, 119-126), (Rykaczewska-Czerwińska M. et al., 

2001, Comp. Biochem. Physiol. Part C, 130, 271-279). This effect is mediated by 

central opioid receptors. The other recently reported effect of LPK and [2-8]-LPK is 

their prominent accumulation in adrenals and cortisol releasing into the blood in rats 

(Rykaczewska-Czerwińska M. et al. 2010, Pharmacol. Reports 2010,62, Suppl. p. 95). 

Interferons (IFNs) are a family of proteins belonging to naturally occurring class 

of glycoproteins named cytokines. IFNs are produced and released in response of the 

host to the presence of different pathogens in the viral or bacterial infection, or of 

tumoral cells. IFNs activate immune system of the host. 

In this report we present results of the study of effect of icv administration LPK 

or [2-8]-LPK on IFN-γ plasma level in rats. Investigation was performed on rats, 

which 7 days earlier were implanted with polyethylene cannulas into the lateral brain 

ventricle (icv). On the day of experiment LPK or [2-8]-LPK dissolved in a constant 

volume of 5μl of 0.9% NaCl were icv injected through implanted cannulas. Control 

group was treated icv with the same volume of 0.9% NaCl. After 2 and 24 hours 

animals were anesthetized and a sample of the blood was taken by heart puncture. 

IFN-γ level was determined in plasma by the method of ELISA. It was found that icv 

administration of LPK or [2-8]-LPK induced significant increase of IFN-γ in rats 

plasma. Pretreatment rats with Naloxone (1mg/kg ip), or with equimolar dose of 

antagonist LPK and of μ-opioid receptors prevented the effect LPK or [2-8]-LPK. 

Thus obtained results evidently proved immunostimulatory effect of both evaluated 

peptides: LPK and [2-8]-LPK. We suppose that these both peptides may be a useful 

tool substances for modulation of production IFNs blood levels in rats. Moreover it 

was revealed that central opioid receptors mediate immunostimulatory effect of LPK 

and [2-8]-LPK. 

At present β-interferons are used in the treatment of multiple sclerosis, an idiopathic, 

chronic, inflammatory diseases of central nervous system in humans. 



O-30 

INVESTIGATIONS ON RESISTANCE OF THE CODLING 

MOTH CYDIA POMONELLA (L.) (LEP., TORTRICIDAE) 

TO INSECTICIDES IN POLAND 

Damian Gorzka, Remigiusz W. Olszak, Michał Hołdaj 

Institute of Pomology and Floriculture. Poland, 96-100 Skierniewice, 

Pomologiczna 18 st. 

The codling moth (Cydia pomonella) is one of the most significant pests 

occurring in apple orchards in all regions of apple tree cultivation. Increasing 

frequency of chemical treatments led to acquisition of pest resistance to a considerable 

number of recommended plant protection products. Since many years of insecticide 

treatments, Cydia pomonella developed, cross and multiple resistance to different 

pesticides in many parts of world. We have a few tools to measure pesticide resistance. 

In our investigations codling moth resistance was tested by analysis of enzymatic 

activity of three complex: mixed function oxidases (MFO), glutathione transferases 

(GST), and esterases (EST). That enzymatic activities, participate in codling moth 

insecticide detoxification. For bioassay investigation 5 field populations of 

C. pomonella from different parts in Poland (pesticides treated orchards) was used in 

comparison with laboratory strain as control. Field materials (adult moths) was 

collected systematically during all vegetative season of 2010 and 2011, using different 

kind of traps. Bioassay were performed using enzymatic activity of three complex 

which participate in codling moth insecticide detoxifications. Resistances to azinphosmethyl, 

diflubenzuron, spinosad, tebufenozide and thiacloprid are significantly 

correlate with mixed function oxidases activity, while increased glutathione 

transferases and reduced non-specific esterases activity is correlate with resistance to 

azinphos-methyl and emamectin, respectively. Conversely, resistances to azinphosmethyl, 

tebufenozide and thiacloprid were negatively correlate with increased 

esterases activity. During now performed 1500 enzimatic analises of MFO, EST GST 

from different field populations. In our investigation we observed high activity 

of esterases (EST) however enzymatic analyses are still in progress. Enzymatic 

activity test is one of innovative, fast and reliable instrument to check insecticide 

resistance in Tortricidae pests. As it is known from literature only a few of the 

registered insecticides against Cydia pomonella L. are still fully effective in areas 

where insecticide resistance has emerged in this pest. 

Keywords: Cydia pomonella, insecticide resistance 



O-31 

THE EFFECT OF SPRING APPLICATION OF 

MICRONUTRIENT (Zn, Ca, U) AND PESTICIDEON 

ABUNDANCE OF EGG AND NYMPHS COMMON 

PISTACHIO PSYLLA, AGONOSCENA PISTACIAE (HEM.: 

APHALASRIDAE) IN PISTACHIO ORCHARDS 

44 

M. A. Samih, M. Rouhani, M. Esmaiilizadeh, M. Fekri 

Department of Plant Protection, Faculty of Agriculture, Vali-e-Asr University, 

Rafsanjan, Iran, e-mail: rouhani_valiasr@yahoo.com 

The common pistachio psylla, Agonoscena pistaciae Burckhardt and Lauterer 

(Hem.: Aphalaridae) is one of the most important pests of pistachio, trees distributed 

in all pistachio producing regions of Iran. Nowadays various pesticides are used to 

control this pest. Extreme spraying against this pest during many years has defeated its 

natural enemies. Moreover, the overlapping of generations has caused its overflow. 

Many herbivorous insects can qualitatively distinguish among host plants, and feed 

and oviposit preferentially on high quality plants. Cultural methods such as crop 

fertilization can affect susceptibility of plants to insect pests by altering plant tissue 

nutrient levels. we studied in relation to varying fertilization levels of N, Ca, Zn 

on common pistachio psylla, A. pistaciae in pistachio orchards based on complete 

randomized blocks with three replications and nine treatments including once-applied 

Zn, Ca, U, UCa, ZnCa, ZnU, ZnCaU, Pesticide (amitraz) and control (distilled water) 

on common pistachio psylla fed on 20-year-old pistachio trees of Ahmadaghaei 

rootstock. The results showed that the effect of nutrition solution on nymph and eggs 

population had a significant difference at %5 level, and the highest measure of control 

on eggs was related to the combination of Pesticide, ZnCa and Zn while the least was 

related to UZn also results showed that the highest measure of control was related 

to the combination of UZn, Ca and UZnCa while the least was related to U and Zn. 

The results indicated that the combination of the elements mentioned had a stronger 

reducing effect on the concentration of pistachio psylla nymphs than that of pesticide 

too. 

Keywords: Agonoscena pistaciae, amitraz, nutrition element, pesticide. 



O-32 

BOMBYX TRANSFORMATION AS A TOOL FOR 

PHYSIOLOGICAL STUDIES 

Duńan Ņitňan 1 ,Ivana Daubnerová 1,2 , Ladislav Roller 1 , Michael E. Adams 3 

1 Institute of Zoology, SAV, Dúbravská cesta 9, Bratislava, Slovakia; 

dusan.zitnan@savba.sk 

2 Department of Genetics, Faculty of Natural Sciences, Comenius University, 

Bratislava, Slovakia 

3 Depts of Entomology & Neuroscience, University of California, 

Riverside, 92521, USA 

Germ-line transformation using various promoters and molecular markers 

became an essential approach to understand complex physiological and biochemical 

processes in living organisms including insects. In spite of remarkable economic and 

medical importance of many pests and pathogen vectors (e.g. moths, beetles, flies, 

mosquitoes), routine transformation has been performed in a single species 

(Drosophila), while it is very difficult or impossible in other species. Here we will 

discuss our progress in Bombyx transduction and transformation using the baculovirus 

and piggyBac systems. We used these systems to understand hormonal regulation 

of neuropeptide genes and receptors, as well as to determine their physiological 

functions in the central nervous system and endocrine glands. To drive expression 

of molecular markers and receptors in specific neurons or endocrine cells we identified 

numerous neuropeptide promoters from the Bombyx genome databases and cloned 

neuropeptide genes. The commercially available baculovirus expression system (Bacto-Bac, 

Invitrogen) and SF9, SF21 cell lines have been employed to evaluate the 

specificity of these promoters in Bombyx. The cassettes containing identified 

promoters and molecular markers, receptors or transcription factors have been 

subcloned into piggyBac vectors. Injections of these constructs into fresh eggs resulted 

in production of transgenic Bombyx lines showing specific expression of various 

markers/receptors in specific peptidergic neurons on the CNS, endocrine Inka cells, 

or corpora allata. Physiological and behavioural consequences of overexpression 

of several peptide receptors in different cell types will be discussed. 

This work was supported by a Slovak grant (VEGA 2/0132/09) and NIH, USA (GM 67310). 



O-33 

ENTOMOPHAGY AND ENTOMOTHERAPY IN NORTH- 

EAST INDIAN TRIBALS 

46 

Meyer-Rochow V.B. 1 , Chakravorty J. 2 

1 School of Engineering & Science, Jacobs University, D-28759 Bremen, Germany 

b.meyer-rochow@jacobs-university.de 

2 Dept. of Zoology, Rajiv Gandhi University, Arunachal Pradesh 791112, India 

At least 80 species of insects, in particular Odonata, Orthoptera, Isoptera, 

Hemiptera, Homoptera, Coleoptera, Lepidoptera, and Hymenoptera, are being used by 

North-East Indian tribals of Nagaland and Arunachal Pradesh as food. Twelve species, 

raw or boiled, find applications in treating stomach disorders, coughs and colds, skin 

allergies, boils, malaria, blood pressure anomalies, scabies (in case of humans) and 

foot and mouth disease of cattle. One widely consumed insect is the ghondibug 

Aspongopus nepalensis, but excessive consumption of this insect can result 

in symptoms akin to anxiety, reminiscent of reports of seasonal ataxia, caused by the 

consumption of African silkworm (Adamolekun et al. 1997) and the use of Coridius 

(Aspongus) chinensis as an aphrodisiac known as “chushan” and “hai tao chung” 

in China (Hoffmann 2004). Since lipid amounts in the ghondibug were higher than 

those of most other insects (crude lipids made up 38.35%), we examined the fatty acid 

content of this species and found MUFAs to represent up 56.1% of the total fat 

content. Oleic and palmitic acids (46.41% and 32.32% of total fat) were the two major 

fatty acids in this insect‟s oil. The latter also contained moderate amounts of stearic, 

palmitoleic, and linoleic, myristic, eicosenoic, and linolenic acids. A. nepalensis was 

rich in Na: 1.02%, K: 0.35%, Ca: 0.12%, Mg: 0.16% and, moreover, contained 

considerable amounts of Fe, Cu, Zn and Mn. With regard to vitamins, A was most 

abundant (34.38 µg/ 100g); C, D, E, and those of the B-complex were present, but less 

abundant. It is obvious that due to population increases the resources of the forest, 

including food insects, can become over-exploited; this has apparently happened 

already in some parts of North-East India. Many of our informants indicated that it has 

become increasingly more difficult to collect useful insect species. We, therefore, see 

an urgent need to assess insect biodiversity and the role of ethno-entomology together 

and not separate from each other. On the one hand, it is desirable to see practices 

of entomophagy and entomotherapy not disappear (at least not before they have been 

thoroughly documented); on the other hand, we need to protect the biodiversity of the 

region and conserve the valuable insect resources found there for posterity. 

References 

Adamolekun B. et al. (1997) Metabol. Brain Dis. 12, 251. 

Hoffmann, H.J. (2004) Heteropteron 18, 19. 



O-34 

CONTROL OF INSECT DIGGESTION: ARE 

ADIPOKINETIC HORMONES INVOLVED? 

D. Kodrík 1,2 , K. Vinokurov 1 , A. Tomčala 3 , R. Socha 1 

1 Institute of Entomology, Biology Centre, CAS, České Budějovice, Czech Republic 

E-mails: kodrik@entu.cas.cz, orchesia@seznam.cz, socha@entu.cas.cz 

2 Faculty of Science, South Bohemian University, České Budějovice, Czech Republic 

3 Institute of Organic Chemistry and Biochemistry, CAS, Praha, Czech Republic 

E-mail: a.tomcala@centrum.cz 

A representative of herbivorous insects, the firebug Pyrrhocoris apterus 

(Heteroptera: Pyrrhocoridae) served as a model species for biochemical 

characterization of midgut digestive processes that constituted a baseline for a study 

of hormonal regulation of insect digestion. The analysis of the midgut content revealed 

acidic conditions with pH in the range 4.6-5.2 for all three studied midgut parts - 

anterior (ANT), middle (MID) and posterior (POS). In accordance with those low pH 

values, the studied digestive hydrolases (peptidase, amylase and alfa-glucosidase) were 

highly active at the same conditions having optimum at pH 5.5. Lipase activity in this 

acidic pH constituted approximately 50% of the maximum (measured at pH 7.5). 

The study also revealed that the distribution of enzymatic activities along the midgut 

parts was not uniform. P. apterus possesses two adipokinetic hormones - octapeptides 

Pyrap-AKH (Kodrík et al., 2000, Insect Biochem. Mol. Biol. 30, 489) and Peram- 

CAH-II (Kodrík et al., 2002, Peptides 23, 585). A major function of these hormones is 

the control of insect metabolism in stress situations, however, these peptides are 

pleiotropic, with a number of actions attached to their metabolic role (Kodrík, 2008, 

Physiol. Entomol. 33, 171). An injection of 10 pmol Pyrap-AKH into the firebug body 

modified the processes in the midgut within 24 hours. The activity of lipases 

(carboxyesterases) was stimulated predominantly in the MID (5.5 times), while in the 

POS the stimulation was moderate (1.3 times) and in the ANT only negligible. 

The level of lipids was enhanced mainly in the content of first two midgut parts, while 

in the midgut tissue the enhancement was much lower, nevertheless in the MID 

significant, suggesting that the food intake is AKH-affected. The protease activity 

increased significantly in all three midgut parts after the hormonal treatment, but the 

stimulation was the most intensive in the POS (almost 3 fold). Accordingly, the protein 

level was also significantly enhanced, but only in the content of all midgut parts; no 

hormonal effect in the gut tissue was recorded. The AKH-effect on carbohydrate 

digestion was disunited: while the activity of glucosidases seems to be stimulated (in 

the MID about 4 times), activity of amylases and glycogen content was not 

significantly affected. It can reflect a carbohydrate specificity of the firebug food, 

linden seeds, which are rich in sugars and poor in starch. If this phenomenon is a result 

of more or less direct effect of AKH on the enzyme producing of midgut epithelium 

columnar cells, and/or an indirect result of food intake stimulation and the midgut 

filling, is a matter of further investigation. 

The study was supported by project No. P501/10/1215 from the Czech Science Foundation (DK). 



O-35 

DIVERSITY OF BEETLES ASSOCIATED WITH ACACIA 

SENEGAL L. (WILD) IN GUM ARABIC BELT OF SUDAN 

48 

Maymoona Ahmed Eisa 1 , Ingo Brunk 2 

1 Technische Universität Dresden, Institute of Forest Botany and Forest Zoology, 

Tharandt, Germany, 1 Fax: +49 (0) 35203-38-31317 email: 

maymoonaeisa@yahoo.com 

1 University of Kordofan, Department of Crop protection, Faculty of Natural resources 

& Environmental Studies, P.O Box 160 -Elobeid 51111, Sudan 

2 Technische Universität Dresden, Institute of Forest Botany and Forest Zoology, 

Tharandt, Germany email: brunkin@web.de 

Corresponding author: M. A. Eisa 1 

The Acacia senegal (gum Arabic producing tree) serves a variety of valuable 

economic and ecological functions. Due to lack of sound information with respect to 

the ecological requirements of the pest species and the environmental factors a study 

was carried during season 2007 and 2008. Main focus was to observe longhorned 

beetles, (Cerambycidae) on the gum belt in northern Kordofan state-Sudan, but also 

other arthropods on Acacia senegal (gum Arabic) tree were sampled. Collection was 

done using flight interception traps located in El Demokeya reserve forest. Fourteen 

potentially pest species of arthropods were identified to be associated with Acacia 

senegal. Among the Coleoptera seven species of Cerambycidae, four species 

of Buprestidae, two species of Meloidae and one of Curculionidae were found. It was 

recommended to detect the presence of these species on other forest locations 

of Acacia senegal. 

. 

Keywords: Acacia senegal, gum belt, Arthropods, Flight traps, Sudan 



O-36 

TWENTY FIVE YEARS OF AZADIRACHTINS 

Hans E. Hummel 

Chair of Organic Agriculture, Justus-Liebig-University Giessen, Germany 

& Prairie Research Institute, University of Illinois, Urbana-Champaign, USA. 

Hans.E.Hummel@agrar.uni-giessen.de 

Azadirachtins (azas) from Indian neem Azadirachta indica, Thai neem 

A. siamensis, and marrango A. excelsa (Meliaceae) seeds have long been recognized as 

phagorepellents, developmental modifiers and sterilants for arthropods while virtually 

harmless for vertebrates. This prompted, during the second half of the last century, 

a search for the natural products which are responsible for the effects observed. 

It culminated in a preliminary structure proposal by Zanno et al. 1975 of aza as 

a tetranortriterpenoid with the molecular structure of C35H44O16. Between 1985 and 

1987 two laboratories in Germany and England independently and simultaneously 

succeeded in the unequivocal structure determination of aza by NMR studies (Kraus et 

al. 1985) and X-ray crystallography (Broughton et al. 1986, Bilton et al.1987). Veitch 

et al. 2007 finally confirmed the structure of aza by total synthesis. - Today, we know 

a larger family of closely related azadirachtins. One of those compound is known 

as marrangin (mar) and was first found, isolated and identified at the Giessen 

laboratory by Ermel et al. (1991), Doll and Schmutterer (1993), and Kalinowski et al. 

(1993). The biological activity of mar (EC50 = 0.25ppm) significantly exceeds the 

potency of aza (EC50 = 1.6 ppm) in the Mexican bean beetle assay of Epilachna 

varivestis. Mar is also active in mites where Sanguanpong 1992 found an EC50 of 1200 

ppm for the mortality of protonymphs of Tetranychus urticae (Acari: Tetranychidae). 

Biorational compounds of this kind today are sought as sustainable, ecologically 

appealing, renewable, and fully biocompatible replacements for classical toxicants. 

Aza and mar are tolerated in organic agriculture where strict rules admit only about 

half a dozen natural products, among them pheromones, azas and Quassia. In spite 

of the comparatively high price of the commercially available neem products, they 

occupy a growing market share. 

Recently, aza and analogs are also highly welcome in veterinary and human medicine 

(Schmutterer 2002, Kleeberg and Strang 2009). In addition, aza and congeners are 

excellent probes for mechanistic studies in arthropod hormone physiology and 

neuroendocrinology. 

References 

Bilton et al. (1987) Tetrahedron. 43, 2805. 

Broughton et al. (1986) J. Chem. Soc., Chem. Comm. 46-47. 

Doll and Schmutterer (1993) Mitt. Dtsch. Ges. Allg. Angew. Entomol. 8, 775. 

Ermel et al. (1991) J. Appl. Entomol. 112, 512. 

Kalinowski et al. (1993) J. Liebigs Ann. Chem. 1993, 1033. 

Kleeberg and Strang (2009) Proceedings of the 14 th workshop, Wetzlar, Germany. 

Kraus et al. (1985) Tetrahedron Letters. 26, 6435. 

Sanguanpong (1992) PhD Thesis, Justus-Liebig-Universität Gießen. 

Schmutterer (2002) The neem tree Azadirachta indica A. Juss. and other meliaceous plants. 2 nd edition, 

Neem Foundation, Mumbai. 

Veitch et al. (2007) Angew. Chem. Int. Ed. 46, 1. 

Zanno et al. (1975) J. Am. Chem. Soc. 97, 1975. 


50 


O-37 

THE ROLE OF COUMARINS AS INSECTICIDAL 

ACTIVITY AGAINST COWPEA BEETLE, 

CALLOSOBRUCHUS MACULATUS (FAB.) 

(COLEOPTERA: BRUCHIDAE) 

I. A. Ismail *, S. E. El-Gengaihi**; A. Affifi ** 

* Pests and Plant Protection Dept. 

** Medicinal and Aromatic Plants Dept., National Research Centre, Dokki, Cairo, 

Egypt, Email: ismfer44@hotmail.com and also, ismailismail12@yahoo.com 

Cowpea beetle, Callosobruchus maculatus (Fab.) (Coleoptera: Bruchidae), is 

the most important storage pest of cowpea. The quantification of cowpea losses 

through C. maculatus is very desirable. Coumarins possess insecticidal activity more 

frequently in Rutaceae and Apiaceae families. Thus some plants grown in Egypt such 

as murraya, kumquat and celery belonging to Rutaceae and Apiaceae known to contain 

coumarins were extracted, isolated and evaluated as insecticidal activity agents for 

controlling cowpea beetle. The major coumarin from each plant was crystallized and 

identification. Chloroform extract of celery was the most toxic as it induce 100% 

mortality after 96 hours with all the concentrations used. Comparing the mortalities 

induced by the two extracts of the three plants, it was obviously revealed that murraya 

had the least toxic effect with both extracts; chloroform and ethanol while, kumquat 

produced moderate effect. 

Keywords: Callosobruchus maculatus, Coumarins, Insecticidal activity. 

Correspondence: Prof. I.A. Ismail, Pests & plant protection Department, National Research Centre, 

Dokki, Egypt. Email: ismfer44@hotmail.com and also, ismailismail12@yahoo.com 



O-38 

SYSTEMIC RNA INTERFERENCE IN THE DESERT 

LOCUST, SCHISTOCERCA GREGARIA 

N. Wynant 1 , H. Verlinden 1 , B. Breugelmans 1 , J. Vanden Broeck 1 

1 Molecular Developmental Physiology and Signal Transduction, Animal Physiology 

and Neurobiology, K.U.Leuven, Naamsestraat 59, 3000 Leuven, Belgium. 

Niels.wynant@bio.kuleuven.be 

Introduction of dsRNA induces RNA interference (RNAi), a sequence-specific 

inhibition of cellular gene expression at the post-transcriptional level. Thanks to its 

selective and robust effect on gene expression RNAi has become a widely used tool to 

knock down and analyse the function of genes. Several recent studies have shown that 

RNAi may also contribute to novel strategies for selectively controlling agricultural 

pests including a number of insect species. Although components of the intracellular 

RNAi machinery have been found in many different eukaryotes, the systemic nature 

of the RNAi-response, whereby the silencing signal can be spread between cells and 

even passed on to the progeny, seems to be more species-specific. The ability of the 

silencing signal to spread has great implications, both as a means to introduce 

insecticidal dsRNA as well as a „loss of function‟-research tool. It was first thought 

that insects were merely insensitive toward extracellularly administrated dsRNA. Now, 

it has become clear that many insect species can respond in a systemic nature, but not 

all show the same degree of sensitivity. By means of reverse transcriptase real time 

(RT-)PCR we reported a very sensitive and long term systemic RNAi response in the 

desert locust. Although this was the case for all investigated tissues, a clear difference 

in sensitivity could be seen. In addition we used RNAi to silence genes that are 

essential for the survival of this voracious pest insect, after which mortality was 

observed. This was only the case when dsRNA was injected directly into the 

hemocoel, since we could not observe the same degree of silencing when dsRNA was 

orally administrated. In conclusion we state that the desert locust seems to contain the 

essential components for a strong and long lasting systemic RNAi response. 


52 


O-39 

THE ROLE OF TWO PUTATIVE SEROTONIN 

RECEPTORS IN THE PHASE TRANSITION OF THE 

DESERT LOCUST, S. GREGARIA 

Vleugels R. 1 , Verlinden. 1 , Ott SR. 2 , Burrows M. 2 , Blenau W. 3 , Vanden Broeck J. 1 

1 Department of Animal Physiology and Neurobiology, K.U. Leuven, Leuven, Belgium 

2 Department of Zoology, University of Cambridge, Cambridge, UK 

3 Department of Animal Physiology, Universität Potsdam, Potsdam, Germany 

E-mail: Rut.Vleugels@bio.kuleuven.be/ Jozef.Vandenbroeck@bio.kuleuven.be 

The desert locust, Schistocerca gregaria, displays an extreme, density 

dependent phenotypic plasticity between a harmless solitarious and a swarming 

gregarious phase. The phase transition is a multilayered process with remarkable 

changes in several aspects of the physiology and the morphology of the locusts. 

However the first characteristic that changes is their behaviour. Within two to four 

hours of crowding, behaviour switches from a strong reciprocal repulsion in solitarious 

locusts to coherent aggregation and higher activity in gregarious ones. Phase transition 

is the reason why desert locusts can form huge, devastating swarms. Serotonin (5hydroxytryptamin, 

5-HT) was shown to play an important role during the initial 

behavioural changes of phase transition. Since interfering in these early stages may be 

an interesting target for preventing locust swarms, the signal transduction pathways 

of 5-HT need to be clarified. Therefore, we investigated two putative 5-HT-receptors 

of S. gregaria (5-HT1Sg and 5-HT7Sg). The transcript levels of both receptors were 

analysed and compared in long term solitarious and long term gregarious locusts using 

real time qRT-PCR. The expression of the putative 5-HT1Sg is higher in crowdedreared 

locusts as compared to isolated-reared individuals. Isolated-reared locusts 

showed higher expression levels of 5-HT7Sg compared to crowded-reared ones. We 

also investigated the effect of a knockdown via RNA interference of both receptors 

on the behavioural changes during gregarisation. 5-HT7Sg RNAi animals tend to be 

more gregarious, indicating a positive effect of the receptor on the behavioural 

gregarisation. 5-HT7Sg couples positively to the adenlyate cyclase (AC), this may 

indicate that AC activity leads to more gregarious behaviour. Knockdown of 5-HT1Sg 

has a positive influence on the behavioural gregarisation, suggesting that the receptor 

itself has a negative influence on the gregarisation process. Since 5-HT1Sg couples 

negatively to AC, this fits into the hypothesis that higher AC activity leads to more 

gregarious behaviour. This may also indicate that 5-HT1Sg is part of a negative 

feedback mechanism to control phase-related behaviour. 



O-40 

GROWTH INHIBITION AND COMPENSATORY 

EFFECTS OF PACIFASTIN-LIKE INHIBITORS ON A 

PEST INSECT: THE DESERT LOCUST, SCHISTOCERCA 

GREGARIA 

Spit J. 1 , Breugelmans B. 1 Simonet G. 1 , Vanden Broeck J. 1 

1 Department of Animal Physiology and Neurobiology, Zoological Institute 

K.U.Leuven, Naamsestraat 59, B-3000 Leuven, Belgium 

Protease or peptidase inhibitors (PI) are considered as important possible 

defence molecules for future insect-resistant crop plants. PI are an important part 

of the natural defence mechanisms in plants upon herbivory. They are present in seeds 

or can be induced after insect feeding. After consumption they inhibit gut proteases, 

which could lead to amino acid shortage, ultimately resulting in growth inhibition. 

Diverse genes from a variety of plant sources are available and multiple PI genes 

of plant origin were considered promising in the past. However, in many cases 

polyphagous pest insects manage to physiologically adapt quickly to these inhibitors. 

The main reason for the varying degrees of success of peptidase inhibitors as 

biological insecticides is the existence of a yet poorly understood mechanism, which 

allows several pest insects to compensate for PI present in their diet. To study this 

highly flexible respons mechanism and to prolong the inhibitory effect of PI on insect 

growth, a number of measures were taken into account before and during feeding 

experiments on a notorious pest insect, the desert locust, Schistocerca gregaria: (i) 

non-plant PI (i.e. pacifastin-related inhibitors) were used to reduce the effects of coevolutionary 

adaptation of the pest insect digestive system. (ii) Based on the main 

types of digestive enzymes present in the midgut, mixtures of multiple PI with 

different enzyme specificity were selected, ensuring maximal inhibition and (iii) 

digestive peptidase samples were taken during the feeding experiments to study 

compensatory responses. 

Contrary to larvae fed on a diet containing plant-derived PI, a significant growth 

impediment was observed in larvae that were fed a mixture of different pacifastin-like 

PI, illustrating the strength of our strategy to use non-plant PI. Nevertheless, the 

insects physiologically adapted to the PI after a few days, attenuating the growth 

inhibition effect. Moreover, a comprehensive study of the observed compensatory 

responses that occurred during the feeding experiments revealed that S. gregaria 

larvae can respond to PI in their diet in two distinct ways depending on the 

composition/concentration of the PI-mixture. 


54 


O-41 

MODELING AND PREDICTING THE POTENTIAL 

DISTRIBUTION OF FOUR HETEROPTERAN 

PREDATORS IN IRAN 

Samaneh Solhjouyefard 1 , Alimorad Sarafrazi 2 , Mehdi Moeini Minbashi 3 , 

Ali Ahadiyat 1 

1 Department of Entomology, Science and Research Branch, Islamic Azad University, 

Tehran, Iran, e-mail: s.solhjouyefard@srbiau.ac.ir, a.ahadiyat@srbiau.ac.ir 

2 Department of Insect Taxonomy Research, The Iranian Research Institute of Plant 

Protection, Tehran, Iran, e-mail: sarafrazi@iripp.ir 

3 Department of Weed Science Research, The Iranian Research Institute of Plant 

Protection, Tehran, Iran, e-mail: mehdiminbashi@gmail.com 

Prediction and modeling of potential suitable habitats of entomophagous 

species is an important factor in their conservation and further practical application 

in integrated pest management (IPM) programs. In order to assess the influence 

of environmental factors on their potential distribution in Iran, predictive species 

distribution models on four entomophagous species including Nabis palifer, N. 

Pseudoferus, (Nabidae), Orius albidipennis (Anthocoridae) and Geocoris sp. 

(Lygaediae) were obtained using the Maximum Entropy (MaxEnt) method. The 

presence-only data and a set of environmental predictor variables were used. For 

calibrating and testing the accuracy of models, 75% training and 25% testing datasets, 

were used respectively. Environmental datasets consisted of altitude, temperature, 

precipitation layers and their relatives. The area under the curve (AUC) as evaluation 

statistic was also evaluated. The average AUC values for training and testing samples 

ranged between 0.81-0.91 and 0.74-0.86 respectively. According to the obtained maps, 

the prediction outcomes made by Maxent, besides considering the effective factors for 

predicting the distribution of N. palifer (e.g. precipitation seasonality and annual 

precipitation) all suggested the Khorasan-e Shomali, Tehran, Markazi, Hamedan and 

northen Semnan proviences as the suitable areas for this species. North, north-west and 

part of south-west of Iran (e.g. Golestan, Mazandaran, Azarbayjan-e Sharghi 

provinces) were more suitable areas for N. pseudoferus. In terms of this species, 

isothermality and precipitation seasonality included as the effective variables. For 

Orius albidipennis suitable areas included west, north-west, south-west, north-east and 

part of south-east of Iran, altitude and temperature annual range were effective factors. 

Annual precipitation was effective variable for predicting the distribution of Geocoris 

sp. and suitable areas included north, west, south-west and part of north-east of Iran. 

The results could be used as an effective tool for conservation and using the predators 

in further IPM plannings. 



O-42 

EVALUATING INHIBITORY EFFECTS OF SOME 

COMPOUNDS ON DIGESTIVE ALPHA-AMYLSE OF 

MEDITERRANEAN MEAL MOTH ANAGASTA 

KUEHNIELLA ZELLER 

Mohammadi, D. 1 , Farshbaf Pourabad, R. 2 , Jafarlu, M. 3 , Valizade, M. 4 

1 Department of Plant Protection, University of Tarbiat Moallem Azarbaijan, Tabriz, 

Iran, d.mohammadi@ymail.com 




majidjafarlu1983@yahoo.com 

4 Department of agronomy and Plant breeding, University of Tabriz, Tabriz, Iran, 

mvalizadeh@tabrizu.ac.ir 

In this investigation Mediterranean meal moth reared on commercial wheat 

meal in controlled condition and the last male and female larval stages were used for 

inhibitory assays. Alpha-amylase activity measured using specific diagnostic kit (Pars 

Azemon, Iran). The results revealed that effects of different concentrations of ethanol 

on male and female amylase activity were statistically significant and the 

concentration of 96% had the most inhibitory activity. Enzyme activity in the presence 

of different NaCl concentrations in both male and female digestive enzymes were 

affected and with increasing NaCl concentrations enzyme activity decreased. Also 

different concentrations of SDS affected alpha-amylase activity directly and the most 

enzyme activity after control was detected in 1mM and the least activity were detected 

in 5mM of SDS. TRIS affected enzyme activity such as SDS. EDTA affected alphaamylase 

activity differently in sexes and about male insects inhibitory activity in the 

presence of different concentrations of EDTA was not affected. Magnesium Nitrate 

and Potassium Phosphate also affected alpha-amylase activity with increasing their 

concentrations 



O-43 

EFFECT OF LUFOX, INSECT GROWTH REGULATOR, 

ON GRAPE BERRY MOTH, LOBESIA BOTRANA DEN. & 

SCHIFF. (LEPIDOPTERA: TORTRICIDAE) 

56 

Nouraddin Shayesteh* 2 , JafarHosseinzadeh* 1 , Hossein Farazmand 3 , 

Younes Karimpour 1 

1. Department of Plant Protection, Faculty of Agriculture Science, Urmia University, 

Urmia, Iran. Email: Jafar.entomologist@gmail.com, Y.karimpour@mail.urmia.ac.ir 

2. Department of Plant medicine, Mahabad Branch, Islamic Azad University, 

Mahabad, Iran. Email: Profshayesteh@yahoo.com 

3. Department of Agricultural Entomology, Iranian Research Institute of Plant 

Protection, Tehran, Iran. Email: Farazmand@entomology.ir 

*Corresponding authors, E-mail: Profshayesteh@yahoo.com, 

Jafar.entomologist@gmail.com 

The grape berry moth, LobesiabotranaDen & Schiff (Lepidoptera: 

Tortricidae),is a major pest of vineyards in the world and also in Iran. Currently the 

most effective way in controlling of this pest, is using chemical insecticides. Lufox is 

being an insect growth regulator and is a mixture of Lufenuron and Fenoxicarb, that 

both are from insect growth regulator pesticides.In this study the effect of Lufox has 

been compared with Phosalone and that application of insecticides has been done 

against the three pest generations. Different factors including: percentage of infected 

bunches, and percentage of infected berries to grape berry moth recorded.The 

resultsanalysis showed significant differences between the treatments. The average 

number damaged grape berries are each bunch were 0.91in Lufox treatments,whereas 

the average of the damaged grape berries in each bunch in the treatment of Phosalone 

and control were 2.74 and 10.45, respectively. According to the obtained results, 

Lufox effect on controlling of grape berry mothis better than Phosalone. 

Keywords:Lobesiabotrana,Lufox, Phosalone, Insect growth regulator, Pheromone traps. 



O-44 

IMPACT OF THIODON ON THE BEHAVIOUR OF 

PLEXIPPUS PAYKULLI (SAVIGNY ET AUDOUIN, 1827) 

Muhammad Tahir, Malook Bano 

Department of Biological Sciences, University of Sargodha, Pakistan 

hafiztahirpk1@yahoo.com 

Spiders are important biological control agents. In the present study which was 

conducted in District Sargodha, Pakistan, we evaluated the potential effects of Thiodon 

(insecticide) on the survival and avoidance behaviour of one salticids spider e.g., 

Plexippus paykulli. For residual testing, filter papers (6 x 12 inches) were dipped in 

150 ml of solution of Thiodon and dried at room temperature. Control spiders were 

exposed to the filter papers dipped in simple water. Mortality was assessed both in the 

treated and control cohort after 24 hours. Plexippus paykulli was found to be 

susceptible to Thiodon. To record the avoidance behaviour Watmann filter papers 

(12X 9 inch) were used. Filter paper was divided into two halves and one half of the 

filter paper was dipped in Thiodon solution while other half in the distilled water. 

Spiders were released on the filter papers and time spend by each spider on herbicide 

or water treated part was recorded. There was significant difference in the time spent 

by Plexippus paykulli on the insecticide or water treated part of filter paper. 

Keywords: Thiodon, spiders, behaviours 


58 


O-45 

BIOLOGY OF ACANTHIOPHILUS HELIANTHI ROSSI 

(DIPTERA: TEPHRITIDAE): A SAFFLOWER PEST OF 

SOUTHERN IRAN 

Saeidi, K, Nur Azura, A 

Department of Plant Protection, Faculty of Agriculture, University Putra Malaysia, 

43400UPM, Serdang, Malaysia 

Corresponding Author: karim Saeidi, Department of Plant Protection, University 

Putra Malaysia, 43400UPM, Serdang. Fax no: +603-86560698, Tel no: +603- 

89467232,email: saeedi391@yahoo.com 

The safflower fly is one of the most important pests of safflower in Iran. Losses 

caused by larval feeding leads to disrupted plant activities, reduction in flower buds, 

and, ultimately, to decreased quality and quantity of crop. This study was conducted 

on Acanthiophilus helianthi Rossi (Diptera: Tephritidae) at the Gachsaran Agricultural 

Research Station in Iran from November 2008 to July 2009. Sampling was performed 

weekly using a two-stage cluster sampling method. For the life cycle studies, the 

infected flower heads were collected from an experimental field plot and were 

developed in cages (160×160×100 cm) from egg to adult under laboratory 

conditions(27±2°C; relative humidity, 60%; and 16/8 L:D).The results showed that the 

first adults emerged gradually in mid April 2009. Female A.helianthi had a preoviposition 

period of 5.8 ± 1.0 days and the average fecundity was 27 ± 3.2 eggs. The 

Eggs were laid in the bracts of flower heads singly or in clusters of 3-18. The 

Incubation period was 3.8 ± 0.6 days under field conditions and 3.4 ± 0.6 days under 

cage conditions. The egg dimensions were 1.19 × 0.19 mm. Three larval instars 

occurred, and the larval phase was 7-10 days. The mean body dimensions (L × W) of 

larvae were 4.79 × 1.71mm. Pupa was coarctate and become sluggish before last molt. 

The pupal period was 7.5 days (range,6-9 days). The mean pupal dimensions of male 

and female cocoons were 3.95 × 1.42 and 4.55 × 2.27 mm, respectively. The flies 

spent their entire lifespan from egg to adult inside the flower heads of safflower plants. 

Males emerged earlier than females, but the longevity of the adult females (12 ± 3.0) 

was significantly greater than that of males (8 ± 1.0). Analysis of aggregated male and 

female sampling data showed that the sex ratio was 1:1.28 

Key words: Acanthiophilus helianthi, Safflower fly, Biology, Safflower, Iran 



O-46 

INSECTICIDAL PEPTIDES 

Neil Audsley, Rachel Down, June Matthews 

Food and Environment Research Agency, Sand Hutton, York, YO41 1LZ, UK 

E mail: neil.audsley@fera.gsi.gov.uk 

Insect peptide hormones regulate virtually all physiological processes in 

insects, as well as behaviour. These peptides act by binding to receptors on cell 

membranes, which trigger intracellular changes, leading to a physiological or 

behavioural response. The peptide and/or receptor can be identified then manipulated 

to have detrimental effects on the insect. 

We are currently investigating the regulation of feeding and/or reproductive behaviour 

in aphids, beetles and flies as targets for controlling pest populations. 

Both (C-type) allatostatins and myosupressins suppress feeding activity resulting in an 

increase in mortality when continually fed to aphids or injected into adult house flies, 

but have no effects when injected into beetles or cabbage root flies. The mechanisms 

of action of these peptides in these insects are currently unclear, but it is possible they 

are acting on gut motility. 

In the cabbage root fly (Delia radicum) we have demonstrated that male accessory 

gland extracts suppress mating when injected into virgin female flies. We are currently 

in the process of identifying the factor(s) involved. 



POSTERS



P-1 

THE EFFECT OF GLIOCLADIUM CATENULATUM J1446 

TO THE RESPIRATION RATE OF THE BUMBLE BEE 

BOMBUS TERRESTRIS 

62 

R. Muljar 1 , M. Mänd 2 

1,2 Department of Plant Protection, Estonian University of Life Sciences, Tartu 51014, 

Estonia, e-mail: riin.muljar@emu.ee 

Increasing use of agrochemicals has led to pesticide residues in food and 

environment, has caused resistance problems in the control of many plant pests and 

diseases. An alternative to chemical spraying is to use bees to disperse biological 

control agents, however, the effects of such biocontrol agents to bumble bees, which 

are important pollinators of agricultural and natural ecosystems, are relatively little 

studied. Prestop Mix is a biofungicide that contains spores of the naturally occurring 

parasitic fungus Gliocladium catenulatum J1446. There is no information about the 

effects of G. catenulatum to the respiratory system of bees, which is one of the most 

vulnerable targets of most contemporary pesticides. Therefore our goal was to study 

the effect of G. catenulatum J1446 to the respiration rate of bumble bees. We 

conducted laboratory experiments with commercially produced Bombus terrestris 

colonies. For the oral treatments the test bumble bees were fed for three weeks with a 

mix of pollen, sugar solution and the Prestop Mix preparation, whereas the control 

bees were fed with pollen and sugar solution only. For the contact treatments the test 

bumble bees were dusted with Prestop Mix powder, whereas the control bees remained 

untreated. An infrared gas analyzer (Infralyt-4, VEB, Junkalor, Dessau) was used to 

measure the respiration rate of test and control bees by recording the amount of CO2 

release (VCO2 ml h -1 ). Preliminary results of the laboratory tests show that dusting 

bumble bees with the Prestop Mix powder lowered the respiration rate of treated bees, 

whereas feeding bumble bees with the Prestop Mix preparation had no effect on the 

respiration rate of the treated bees. 

- POSTERS -


P-2 

MODULATION OF THE PARACELLULAR PATHWAY IN 

THE LEPIDOPTERAN LARVAL MIDGUT 

B. Diamante 1 , I. Castelli 1 , F. Pennacchio 2 , B. Giordana 1 , M. Casartelli 1 

1 Dipartimento di Biologia, Università degli Studi di Milano,via Celoria 26, 20113 

Milano, Italy. E-mail: barbara.diamante@unimi.it; morena.casartelli@unimi.it; 

2 Dipartimento di Entomologia e Zoologia Agraria “Filippo Silvestri”, Università di 

Napoli “Federico II”, via Università 100, 80055 Portici, Italy 

Control of insect pests is still mainly performed using broad spectrum chemical 

pesticides even though the limits of this strategy are presently well known: the 

reduction in pest population is associated with an unfavourable alteration of food 

quality and safety, with a strong impact on non-target species and the rise of a 

widespread resistance in target insects. Therefore, significant research efforts are 

devoted to the development of innovative technologies based on the use of biological 

control agents, which include small organic molecules and peptide or protein toxins, 

deriving from plants and insect natural antagonists. For an effective oral delivery of 

these bioinsecticides, it will be crucial to develop strategies to facilitate their passage 

through the midgut barriers. They can reach the haemocoel either through the cellular 

pathway, crossing the two polarized plasma membranes of the epithelial cells, or 

through the paracellular pathway, along the aqueous channels formed by the junctional 

complexes, according to the physico-chemical and biological properties of each 

molecule. 

The insect midgut paracellular pathway is particularly interesting for the delivery 

of bioinsecticides of low molecular weight targeting hemocelic receptors. We have 

previously demonstrated that the permeability of the septate junction (SJ) of 

lepidopteran larval midgut can be modulated by a precise regulation of cytosolic Ca 2+ 

concentration. We have now studied one of the signaling pathway that may lead to the 

intracellular release of Ca 2+ responsible for the increase of SJ permeability. We used 

C10, a medium-chain fatty acid, known to modulate mammalian tight junctions by 

activation of a Ca 2+ ions-mediated intracellular signaling pathway. Addition of C10 to 

the luminal side of lepidopteran larval midguts isolated in conventional Ussing 

chambers, causes a decrease of the transepithelial shunt resistance, i.e. an increase of 

the junction ion conductance, and an increase of the paracellular fluxes of proctolin 

and fluorescein, two organic molecules that cross the epithelium exclusively by the 

paracellular route. Our data show that C10 triggers an IP3- dependent signaling 

cascade by activation of phopholipase C. As a result, Ca 2+ is released from the 

intracellular stores and a calmodulin-dependent kinase is activated. This event leads to 

the phosphorylation of the myosin light chain by myosin light chain kinase. Myosin 

light chain phosphorylation presumably causes a modification of the cytoskeleton 

organization connected to the SJ and thereby an increase of the paracellular 

permeability to ions and small organic molecules. 

- POSTERS - 63

64 


P-3 

THE STRATEGIES ADOPTED BY A DENSOVIRUS TO 

CROSS THE INSECT MIDGUT BARRIER 

I. Castelli 1 , B. Diamante 1 , AS. Gosselin Grenet 2 , M. Ogliastro 2 , M. Casartelli 1 

1 Dipartimento di Biologia, Università degli Studi di Milano, Via Celoria 26, 20113 

Milano, Italy. e-mail: ilaria.castelli@guest.unimi.it; morena.casartelli@unimi.it 

2 Biologie Intégrative et Virologie des Insectes UMR 1231 INRA-Université 

Montpellier II, Place Eugéne Bataillon, Bat. 24, cc101, 34095 Montpellier Cedex 5, 

France 

Densoviruses (DNVs) are parvoviruses highly pathogenic for arthropods, 

mostly insect at larval stages, including agronomical pest and insect vector borne 

disease. They have limited host range and are not pathogenic to vertebrates, 

characteristic that make them particularly interesting as potential pest control agents 

alternative to chemical pesticides. As a model, we studied the Junonia coenia 

Densovirus (JcDNV) infection of the Lepidopteran pest Spodoptera frugiperda. In 

natural condition JcDNV infection is initiated after ingestion of viral particles. The 

virus does not replicate in intestinal cells, so the midgut epithelium represents a barrier 

that the virus has to overcome to reach the internal target tissue. To understand the 

early step of infection, we first analyzed the JcDNV entry in midgut cells in culture. 

We showed that JcDNV enters specifically columnar cells, but not stem and goblet 

cells, the three cells types that form the midgut epithelium. Mounting isolated midgut 

in a Ussing chamber in the presence of JcDNV in the luminal compartment, we 

showed that after 10 min of incubation viral particles are detectable in the cytoplasm 

of midgut cells and after 30 min or more of exposition the virus appears in the 

intercellular spaces. We also demonstrated that JcDNV internalization induces a raise 

of cytosolic Ca 2+ concentration in columnar cells, an intracellular mediator that causes 

an increase of the epithelium permeability through the paracellular pathway. We 

effectively verified, in Ussing chamber, that JcDNV causes a significant decrease of 

the paracellular electrical resistance, i.e an increase of the septate junction (SJ) 

permeability to ions. Using the same apparatus we also observed that after 10 min of 

incubation the amount of virus able to cross the epithelium is lower compared to that 

measured after 30 min. Therefore our results indicate that the internalization of JcDNV 

into midgut cells causes the raise of intracellular calcium concentration and, 

consequently, the increase of the permeability of the paracellular route, the pathway 

used by the virus to cross the midgut epithelium to reach in a massive amount the 

internal target tissues. To characterized the mechanism involved in JcDNV 

internalization into midgut cells, isolated midguts were mounted in a Ussing chamber 

and incubated with the virus in the absence or in the presence of drugs able to inhibit 

specific endocytic pathway. Our results show that clathrin-coated vesicle formation 

and lipid rafts-dependent endocytosis are essential for JcDNV entry into columnar 

cells. Using specific molecular markers for vesicle trafficking we also demonstrated 

that early and late endosomes are involved in JcDNV intracellular trafficking. 

- POSTERS -


P-4 

AN ATTEMPT OF NATURAL SUPERVISION 

OF AN ABUNDANCE OF INSECT PLANT FEEDERS 

ON THE WHEAT FIELD WITHOUT APPLICATION 

OF PESTICIDES 

A. Semenov 

Department of Entomology, Moscow State University, Moscow, Russian Federation, 

SashaSem7@yandex.ru 

The gradation of grievous plant feeders in some certain cases can be prevented 

by creating zones with other plants on the field which are able to increase the species 

diversity of arthropods. In this case the ecosystem can become more stable. 

The purpose of the study was to reveal how zones with an alfalfa influence on the 

insect complex of the wheat field. 

Our work was held in Stavropol Territory on the field of the common wheat 

(Triticum aestivum L.). We seeded zones with an alfalfa (Medicago sativa L.) which 

crossed the field. These zones were parallel, with 200 meters of distance. The width of 

zones was about 3 meters. 

The data which we get from an entomological mowing revealed that the 

influence of an alfalfa zones can be observed not further than 100 meters from an 

alfalfa and is expressed in the variation of the species composition and abundance of 

some certain species. Variation in the ground fauna shows itself well when moving 

away alfalfa zone at least on 10 m. 

The alfalfa zones influence well both on predatory insects and on grievous 

plant feeders. We showed the tendency to increase abundance for certain species 

(Zabrus tenebrioides G-Z, Eurygaster intergriceps Put.), for other species – to 

decrease (Philaenus spumarius L.). It depends on increasing of the distance to the 

alfalfa zone. 

- POSTERS - 65


P-5 

LC/MS AND GC FID TECHNIQUES EMPLOYED IN THE 

ANALYSES OF LIPIDS INVOLVED IN INSECT 

METABOLIC AND ADAPTATION PROCESSES 

66 

Tomčala A. 1 , Kindl J. 1 , Schneedorferová I. 3 , Kodrík D. 2 , Valterová I. 1 

1 Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech 

Republic, Flemingovo nám. 2, 166 10 Prague, Czech Republic, 

2 Institute of Entomology, Biology Centre, Academy of Sciences of the Czech Republic, 

Branišovská 31, 370 05 České Budějovice, Czech Republic 

3 Czech Technical University in Prague, Faculty of Biomedical Engineering 

a.tomcala@centrum.cz 

Insects represent a very useful experimental model in various branches of 

biological research. The investigation is driven by economic importance of many 

insect species, and also by biological features of insects as model organisms such as 

short period of reproduction, easy breeding and manipulation and, in particular, the 

minimal regulatory requirements as compared with the management of vertebrates. 

Here we present our robust and efficient LC/MS (liquid chromatography/mass 

spectrometry) and GC/FID (gas chromatography/flame ionisation detection) methods 

for the determination of physiologically important lipids in insects. Target molecules 

are polar (glycerophospholipids - GPL) and non polar lipids (diacylglycerols - DG and 

triacylglycerols - TG). Combination of the LC/MS data with the subsequent GC fatty 

acid (FA) analysis enables complete structural elucidation of particular lipid species 

including their fatty acid composition. 

The methodology was applied to studies of the chill tolerance of the invasive 

heteropteran species Oxycarenus lavaterae. The fields animals were collected during 

one year to reveal a fluctuation of physiological characteristics related to chill 

tolerance with emphasis on the restructuring of the GPL composition. A relationship 

between the course of temperature changes and GPL characteristics were evaluated by 

the PCA (principal component analysis). These data revealed a positive correlation 

between the low temperature and the GPL membrane composition. The GC/FID 

technique was also employed for the polyol quantitative and qualitative investigations. 

Adipokinetic hormone (AKH) treatment studies of the model insect species Locusta 

migratoria and Pyrrhocoris apterus proved the specificity of particular AKHs on 

ahaemolymph composition of the DGs in the experimental insects. Heterelogous assay 

- application of the L. migratoria AKHs into the P. apterus body and vice versa – 

revealed that the AKH action is rather species than AKH specific. 

Biochemical studies involved also a representative of the Bombus genus, where the 

transport of DGs in haemolymph with emphasis to a function of FAs in bumblebee 

lipid metabolism and pheromone biosynthesis was studied. The structural 

determination of particular DGs and TGs was done, however, further more complex 

study is necessary for better understanding of the phenomenon. 

The study was supported by the Czech Science Foundation projects Nos. P501/10/1215 (DK), and 

P502/10/1734 (AT, IV). 

- POSTERS -


P-6 

ELECTROPHYSIOLOGICAL RESPONSE OF THE 

ANTENNAL TASTE NEURONS TO PLANT SUGARS 

AND SALT IN THE CLICK BEETLE AGRIOTES 

OBSCURUS 

E. Tooming, E. Merivee, A. Must 

Institute of Agricultural and Environmental Sciences, Estonian University of Life 

Sciences, 1 Kreutzwaldi street, 51014 Tartu, Estonia, e-mail: ene.vanatalo@emu.ee 

Electrophysiological responses of antennal taste bristles to thirteen 0,01–100 

mmol l -1 plant sugars were tested in the click beetle Agriotes obscurus (Coleoptera, 

Elateridae), an important pest of many agricultural crops. For the first time, the salt-, 

sugar- and mechano-sensitive neurons were identified in the adult click beetles. 

Sucrose and fructose were the two most stimulatory sugars for the sugar-sensitive 

neuron. The firing rates they evoked were concentration dependent and reached 

approximately 30 imp/s at 100 mmol l -1 . The stimulatory effect of arabinose, glucose, 

mannose, maltose and raffinose was approximately by 65–85% lower. Seasonal 

changes in sensitivity of antennal salt- and sugar-sensitive neurons were observed. In 

winter, in hibernating cold reactivated beetles, the taste neurons were considerably 

more sensitive to tested compounds compared to those of reproductively active beetles 

in summer. Tested active plant sugars are probably involved in maturation feeding of 

the beetles in spring and in searching for their oviposition site in summer. 

- POSTERS - 67

68 


P-7 

THE GENE, TRANSCRIPT AND PEPTIDE ANALYSES 

OF ADIPOKINETIC HORMONE IN THE PEA APHID, 

ACYRTHOSIPHON PISUM 

P. Jedlička 1 , V. Steinbauerová 2 , P. Ńimek 3 

1 Department of Biochemistry and Physiology, Institute of Entomology, Biology Centre 

AS CR, v.v.i., České Budějovice, Czech Republic, e-mail: jedlicka@entu.cas.cz 

2 Faculty of Science, University of South Bohemia, České Budějovice, Czech Republic, 

e-mail: v.steinbauerova@post.cz 

3 Laboratory of Analytical Biochemistry, Biology Centre AS CR, v.v.i., České 

Budějovice, Czech Republic, e-mail: psimek@bclab.eu 

The aphids are known to be important plant phloem-sucking pests, and detailed 

knowledge of their sugar metabolism can effectively contribute to their control 

management. The insect neuropeptides belonging to the Adipokinec Hormone/Red 

Pigment Concentrating Hormone family seem to be good target for this purpose 

(Kodrík D., 2008, Physiol. Entomol. 33, 171). Therefore the main goal of this study 

was to identify a coding region of the Acypi-AKH gene and to analyse its transcription. 

Furthermore, a potential role of this hormone when applied into the aphid‟s body was 

studied as well. 

Molecular study of the Acypi-AKH involved analysis of the gene expression and 

detection of the genomic sequence. Using the 3' RACE PCR method we revealed 

multiple polyadenylation sites in the Acypi-AKH transcripts. Moreover, we compared 

the transcript levels of Acypi-AKH among different morphs. Studying the genomic 

sequence we detected a large intron within the coding region which is over 2.8 kbp 

long. The identification of active form, the Acypi-AKH decapeptide, was subjected by 

the LC/MS and +ESI tandem mass spectrometry. The amount of Acypi-AKH was 

estimated to be 0.79 fmol per apterous female. Applying of a synthetic Acypi-AKH 

into the apterous females we recorded significant increase in concentration of nonreducing 

sugars, mainly glucose, in the whole body extracts. To evaluate this finding 

more accurate we analyzed changes in amount of specific sugars (glucose, trehalose, 

fructose etc.) in the haemolymph and guts of the Acypi-AKH treated aphids using the 

LC/MS. The effect of Acypi-AKH on sugar mobilization in A. pisum virginoparas was 

compared with other “carbohydrate-fuelled” insect species. 

This study was supported by the grant No. 522/09/P382 from the Czech Science Foundation 

- POSTERS -


P-8 

TOXICITY AND PHYSIOLOGICAL EFFECT OF FOUR 

ESSENTIAL OILS OF LABIATAE ON GLYPHODES 

PYLOALIS WALKER (LEP: PYRALIDAE) 

Elham Yazdani, Jalal Jalali Sendi*, Jalil Hajizadeh, Mohammad Ghadamyari 

Department of Plant Protection, College of Agriculture, University of Guilan, Rasht, 

Iran, E-mail: jjalali@guilan.ac.ir 

Essential oils of four plants belonging to family Labiatae were investigated on 

toxicity and physiological aspects of mulberry pyralid Glyphodes pyloalis Walker in 

controlled condition (24±2ºC, 75±5 RH and 16:8 D:L). Acute toxicity and the effect of 

LC10, LC30 and LC50 on feeding efficiency and biochemical indices have been 

performed by leaf disk method on 4 th instar larva. LC10, LC30 and LC50 were estimated 

0.107%, 0.188% and 0.279% for Thymus vulgaris L., 0.31%, 0.705% and 1.24% for 

Lavandula angustifolia Mill, 0.77%, 1.189% and 1.595% for Rosmarinus officinallis 

L. and 0.67%, 1.173% and 1.728% for Origanum vulgare L. respectively. The results 

showed the highest toxicity for T. vulgaris and the lowest one for O. vulgar. The effect 

of sublethal dose LC30 was investigated on relative growth rate (RGR), efficiency of 

conversion of ingested food (ECI), efficiency of conversion of digested food (ECD), 

approximate digestibility (AD) and consumption index (CI) and found to be reduced 

compared with the controls. The effect of the essential oil on total protein, 

carbohydrate, lipid and α-amylase was reduced compared with the controls. Similarly 

protease, lipase, sterases and glutathione S- transferase showed significant differences 

with the controls. Hence, it is concluded that the used concentrations of the essential 

oils not only has toxicity and deterrence on G. pyloalis but also has nonreversible 

effect on the metabolism of biochemical compounds of the test insect. 

Keywords: Mulberry pyralid, Thymus vulgaris, Lavandula angustifolia, Rosmarinus officinallis, 

Origanum vulgare. 

- POSTERS - 69

70 


P-9 

MYOINHIBITORY EFFECTS OF SHORT 

NEUROPEPTIDE F LED-NPF-I ON THE OVIDUCT 

OF BEETLES 

Paweł Marciniak 1* , Lidia Rogalska 1 , Joanna Pacholska-Bogalska 1 , Monika Szymczak 1 , 

Mariola Kuczer 2 , Danuta Konopińska 2 , Grzegorz Rosiński 1 

1 Department Of Animal Physiology and Development, Adam Mickiewicz University, 

Umultowska 89 str., 61-614 Poznan, Poland 

2 Faculty of Chemistry, Wrocław University, Joliot-Curie 14 str., 50-383 Wroclaw, 

Poland, *pmarcin@amu.edu.pl 

Family of short neuropeptide F (sNPF) consists of peptides (typically 3-10 

amino acid residues) isolated from several insect species, including flies, mosquitoes, 

moths, cockroaches and beetles. All of them share common C-terminus sequence – 

RFLRFamide and are recently distinguished from neuropeptides F (typically 36 amino 

acid residues). sNPF are referred as hormones with pleiotropic physiological activities, 

involved in feeding, growth, regulation of locomotion and circadian clock, water 

homeostasis and possibly in learning and olfaction. 

In beetles, sNPF have been characterised in two species Tribolium castaneum 

and Leptinotarsa decmlineata. In L. decemlineata, two peptides Led-NPF-I 

(ARGPQLRLRFa) and Led-NPF-II (APSLRLRFa) were isolated from head extract 

and proposed as diapause regulators. Our previous study showed that Led-NPF-I 

inhibits heart contractions frequency in two beetles Tenebrio molitor and Zophobas 

atratus. Here we report effects of this peptide on the oviduct of the beetles. Led-NPF-I 

inhibited frequency contractions of the oviduct of both beetles. It showed dosedependent 

and species-specific effects. The T. molitor oviduct was inhibited at the 

concentration range 10 -12 -10 -8 M, wheras a threshold concentration for an observable 

effect on the Z. atratus oviduct was 10 -10 M. In both species, the maximal observable 

effect was on the same level and amounted to 20% of inhibition. In addition, several 

analogues were tested, in order to establish agonists and antagonists of sNPF receptor 

in the oviduct muscles. 

- POSTERS -


P-10 

NEW ALLOFERON ANALOGUES AND THEIR IN VITRO 

ANTIVIRAL EFFECT 

Kuczer Mariola 1 , Majewska Anna 2 , Midak-Siewirska Anna 2 , Zahorska Renata 2 , Łuczak 

Mirosław 2 , Konopińska Danuta 1 

1 Faculty of Chemistry, University of Wroclaw, Wroclaw, POLAND; 

2 Department of Microbiology, Medical University of Warsaw, Warsaw, POLAND 

Alloferon is tridecapeptides (H-His-Gly-Val-Ser-Gly-His-Gly-Gln-His-Gly- 

Val-His-Gly-OH) found in blood of an experimentally bacteria-challenged blow fly 

Calliphora vicina [1]. This peptide has antiviral and antitumor properties in vitro [1, 2]. 

Continuing our studies on structure/activity relationship in alloferon we performed the 

synthesis of a series of further analogues of alloferon such as: 

H-Tyr-Gly-Val-Ser-Gly-His-Gly-Gln-His-Gly-Val-His-Gly-OH, H-Trp-Gly-Val-Ser- 

Gly-His-Gly-Gln-His-Gly-Val-His-Gly-OH, H-Phe-Gly-Val-Ser-Gly-His-Gly-Gln- 

His-Gly-Val-His-Gly-OH, H-Phg-Gly-Val-Ser-Gly-His-Gly-Gln-His-Gly-Val-His- 

Gly-OH, H-Phe(p-Cl)-Gly-Val-Ser-Gly-His-Gly-Gln-His-Gly-Val-His-Gly-OH, H- 

Phe(p-OMe)-Gly-Val-Ser-Gly-His-Gly-Gln-His-Gly-Val-His-Gly-OH, H-Gly-Val- 

Ser-Gly-His-Gly-Gln-His-Gly-Val-His-Gly-OH, H-Val-Ser-Gly-His-Gly-Gln-His- 

Gly-Val-His-Gly-OH, H-Ser-Gly-His-Gly-Gln-His-Gly-Val-His-Gly-OH, H-Gly-His- 

Gly-Gln-His-Gly-Val-His-Gly-OH, H-His-Gly-Gln-His-Gly-Val-His-Gly-OH, H-Gly- 

Gln-His-Gly-Val-His-Gly-OH, H-Gln-His-Gly-Val-His-Gly-OH, H-His-Gly-Val-His- 

Gly-OH, H-Gly-Val-His-Gly-OH. 

Peptides were synthesized by the standard solid phase method. The cytotoxic 

properties of investigated peptides were evaluated using Vero, LLC-MK2 and HEp-2 

cell lines. The cytotoxicity was determined by the MTT (3-[4,5 dimethylthiazol-2-yl]- 

2,5 diphenyl tetrazolium bromide) assay. In further experiments the antiviral activity 

was assessed in respect to HHV-1MC, the wild-type HHV-1, and 971 PT Coxsackie 

type B2, the wild type Coxsackie type B2. We found that most of the evaluated 

peptides presented antiviral activity. Moreover, all the peptides did not show cytotoxic 

activity against cell lines. 

References 

1. Chernysh S, Kim S. I, Bekker G, Pleskach V. A, Filatova N. A,. Anikin V. B, Platonov V. 

G, Bulet P. PNAS 99: 12628-12632, 2002 

2. Kuczer M, Dziubasik K, Midak-Siewirska A, Zahorska R, Łuczak M, Konopińska D. J. 

Pept. Sci. 16:186-92, 2010 

Acknowledgements 

This work was supported by the Polish Ministry of Science and Higher Education (Grant No. N N204 

085638), the University of Wroclaw (Grant No. 2237/W/WCh/09) and by the Medical University of 

Warsaw grants AM20/WA and W2/2008–200 

- POSTERS - 71

72 


P-11 

OXICITY OF PHYTOHEMAGGLUTININ ( PHA) 

TO THE GRAIN APHID SITOBION AVENAE Fabricius - 

MODE OF ACTION 

Sprawka I., Czerniewicz P., Goławska S., Sytykiewicz H., B. Leszczyński 

Department of Biochemistry and Molecular Biology, Siedlce University of Natural 

Sciences and Humanities, Siedlce, Poland, e-mail: isprawka@uph.edu.pl 

The insecticidal activity of plant lectins against a wide range of insect species 

have been intensively studied. Understanding the mechanism of the toxicity of lectins 

is one of performed aspects. 

In the present research, the first step was determine the effect of phytohemagglutinin 

(PHA) on the development, fecundity and mortality of grain aphid. Then, the 

following aspects of PHA toxicity mechanisms in the insects were investigated: (1) the 

effect of PHA lectin on the activity of such enzymes as: - and - glucosidases, 

alkaline (AkP) and acid (AcP) phosphatases involved in the metabolism of sugar and 

phosphorus respectively and (2) the effect of PHA lectin on the feeding behavior of an 

adult apterae aphids. 

The all experiments were conducted under laboratory conditions, with an application 

of liquid diets or sucrose-agarose gels of various PHA lectin concentrations. 

The PHA lectin added into the liquid diet increased the pre-reproductive period, 

mortality of S. avenae, the time of generation development and decreased its 

fecundity and the intrinsic rate of natural increase. The aphids fed on sucrose-agarose 

gels with higher concentrations of PHA lectin exhibited no phloem-feeding behavior. 

Total number of the gel penetrations was reduced and duration of the pathways was 

increased. In additions, activity of -glucosidase and alkaline phosphatase of adult 

apterae exposed to PHA was reduced. 

The results indicate that the insecticidal activity of PHA on S. avenae may involve 

changes in activity of the enzymes in the midgut and modification of the aphid 

feeding behavior, which means that, the suppression of S. avenae by PHA may involve 

multiple mechanisms 

- POSTERS -


P-12 

THE EFFECTS OF PARAQUAT ON DNA DAMAGE 

IN THE GRASSHOPPER, CHORTHIPPUS BRUNNEUS 

LARVAE 

M. Augustyniak, Ł. Nocoń, P. Łaszczyca, A. Kędziorski, P. Migula 

Department of Animal Physiology and Ecotoxicology, University of Silesia, 

Katowice, Poland, e-mail: pawel.migula@us.edu.pl 

Genotoxic changes are considered as one of the most detrimental effects of 

pesticide use in non-target organisms. Repair of DNA damage may require substantial 

energy expenses and unrepaired damage may cause various functional disturbances 

affecting fecundity, survival and overall fitness of individuals within a population. 

This may result in decline of susceptible populations and affect biological diversity of 

the ecosystem. Hence, it seems reasonable to assess possible genotoxic potential of 

pesticides in order to control their deleterious side-effects. The comet assay seems to 

be well suited for such assessment because it is sensitive, inexpensive and produce fast 

response. 

Paraquat (PQ) is widely used non-selective and fast-acting herbicide. It causes 

production of reactive oxygen species that can manifest their genotoxic activity in 

animals, both invertebrates and vertebrates. The aim of our study was to assess doseresponse 

relationship between a single PQ administration and DNA damage in brain 

cells of hatching grasshopper (Chorthippus brunneus) larvae. 

Cells isolated from cerebral ganglia of one-day old C. brunneus larvae were treated „in 

vivo‟ or 'in vitro‟ with water solutions of PQ in concentrations: 50, 250, 1250 µM PQ, 

and 10, 50, 250 µM PQ respectively. DNA damages, measured by its increased 

content in the comet tail (TDNA) and length of comet tail (TL), were significant in 

PQ-treated groups in comparison with PQ untreated control group. Positive dose-effect 

correlation was stated for TDNA (but not TL) when PQ was applied „in vivo‟. After „in 

vitro‟ PQ administration significant positive dose-effect correlation was stated for TL. 

Moreover, during „in vivo‟ PQ exposure of cells significant decrease of the damage 

was observed after 24 hours. 

Our results clearly pointed out that PQ penetrated through internal barriers and 

directly or indirectly damaged cellular DNA. Time-dependent further decrease of the 

damage suggests involvement of repair mechanisms that probably were induced in 

response to the oxidative stress. 

- POSTERS - 73

74 


P-13 

GLYCOLITIC ACTIVITY IN MIDGUT OF ADULT WESTERN 

CORN ROOTWORM DIABROTICA VIRGIFERA VIRGIFERA 

LECONTE (COLEOPTERA: CHRYSOMELIDAE), A NEW PEST 

SPECIES OF MAIZE IN EUROPE 

Guzik J. 1 , Nakonieczny M. 1 , Tarnawska M. 1 , Bereś P. 2 , Drzewiecki S. 3 And 

Migula P 1 . 

1 Department of Animal Physiology and Ecotoxicology, Faculty of Biology and 

Environmental Protection, University of Silesia, Katowice, Poland, 

e-mail: jguzik@us.edu.pl 

2 Institute of Plant Protection National Research Institute, Poznań, Poland 

3 Institute of Plant Protection National Research Institute, Sośnicowice Branch, 

Sośnicowice, Poland 

One of the factors influencing survival of insects in the environment is effective food 

assimilation. Hence, our knowledge about activity of midgut enzymes of economically 

important pests would be helpful for better protection of crops. In this study we concentrated 

on the Western corn rootworm, Diabrotica virgifera: Coleoptera; Chrysomelidae (WCR), 

recently next to the corn borer (Ostrinia nubilalis: Lepidoptera; Crambidae), most important 

pest species of maize (Zea mays) in Europe. 

In consequence of active feeding of D. virgifera larvae on maize roots losses in maize crops 

are very high. Adult beetles are active feeders of inflorescence and led to their serious damages 

also reducing maize productivity. Generally this does not cause tying the kernels on the cob. 

So far, biochemical studies of digestive enzymes have focused on larval stages and the 

investigations concerning adult beetles are insufficient. 

The diet of adult D. virgifera is rich in starch and other polysaccharides present in cereals. This 

is the reason why in this study we decided to check what enzymatic spectrum is available to 

effective digestion of a specific food of this stage. Effective digestion is one of the important 

steps for further synthesis of vitellogenins and other energetic substrated in developing yolk. 

We examined activities of a group of glycolytic enzymes. Endoglicosidases: -amylase, 

cellulase, chitinase, licheninase, laminarynase – all that are hydrolyzing polysaccharides. Also 

specific glycosidases: - and -glucosidase, - and -galactosidase were examined using a set 

of specific assays based on spectrophotometric determinations. 

Activity of -amylase , responsible for the first phase of the starch digestion was high. Among 

glycosidases activity of -glucosidase, hydrolysing products of the final digestion of starch 

degradation was the highest among all investigated enzymes from this group. 

Optimal pH values for -amylase, -galactosidase and other specific glycosidases: 4.2, 3.8, 

5.0, respectively were established. The optimum range of pH for -amylase, the main 

hydrolytic enzyme in the metabolism of carbohydrates is relatively large. We suppose that in 

adults, as in the larval stage, two isoenzymes with different pH optimum are active. When 

ranked in a decreasing order of activity of endoglycosidases -amylase, then laminarynase, 

licheninase, chitinase and finally cellulase. In this rank amylolytic activity was ca. 200 times 

higher than that of laminarynase. Among specific glycosidases a high activity level 

characterized -glucosidase and -galactosidase. This means that enzymes most active in food 

digestion in the midgut of adult WCR are -amylase, -glucosidase and -galactosidase - 

enzymes important in hydrolysis of starch, products of its digestion and other compounds 

containing β-galactosidic bonds. 

This study was supported by the grant No. N N310 088936 from the Ministry of Science and Higher 

Education in Poland. 

- POSTERS -


P-14 

PROTEASES OF WESTERN CORN ROOTWORM MIDGUT 

(DIABROTICA VIRGIFERA VIRGIFERA LECONTE) AS A 

POTENTIAL TARGET IN THE MAIZE CROP PROTECTION 

Nakonieczny M. 1 , Guzik J. 1 , Tarnawska M. 1 , Migula P. 1 , Skorek A. 1 , Bereś P. 2 , 

Drzewiecki S. 3 

1 Department of Animal Physiology and Ecotoxicology, University of Silesia, Katowice, 

Poland, e-mail: miroslaw.nakonieczny@us.edu.pl 




A new expansionary pest of maize crops – Western corn rootworm – WRC (Diabrotica 

virgifera virgifera) has been noted in Europe since 1992. After almost 20 years of the first 

imagoes finding in maize plantations near Belgrad in Serbia WRC infested the whole Central 

Europe and reached East France, Central Germany, North Italy. 

D. virgifera is one of the most economically important pests in North America where 

estimated losses only in maize agriculture business are calculated for $ 1 billion. So far the 

available data on its biology are based on the research carried out in US. These unusual 

expanding features of WRC are related both with great adaptative potential of Chrysomelidae 

species to new environmental conditions, and rapidly appearing resistance towards new 

generations of classic herbicides. Therefore, the biology of D. virgifera in European conditions 

should be well found, what would allow finding the most effective methods of controlling this 

pest in Europe and preventing economical loss. 

One of the factors which determine insect‟s survival in the environment is ability to 

effective food assimilation. Hence, knowledge about specific enzyme activity of D. virgifera 

midgut in conditions of feeding in European maize varieties or substitute native plants in 

absence of the main host plant is very important. Especially examination of protease activities 

seems to be crucial in the light of new pest control possibilities by protease inhibitors usage. 

Till now our knowledge about WRC and its protease activities is mostly based on studies on 

larvae stages. 

So far in the midgut of examined imagoes presence of endo- and exoproteases was 

confirmed. Among endoproteases, the highest activity was stated for the serine proteinase – 

trypsin. A chymotrypsin activity depends on the substrate. Similarly to other insect species, the 

high activity of chymotropsin was confirmed only for so-called a long chymotrypsin substrate. 

The optimum pH for both endoproteinases was stated between 10.5 and 11.5. The activity of 

cathepsin G, the other serine proteinase, was much lower. Among cysteine proteinases the 

highest activity was obtained for cathepsin L and much lower for cathepsin B and H with pH 

optimum 5.5 and 6.5-7.0, respectively. The activities of other endoproteases: astacin, elastase 

and papain were also detected. Examination of exoproteases (aminopetidase and 

carboxypeptidases) has shown the major role both of them in the food digestion. Their 

optimum pH activity is 7.5. Among carboxypeptidases, the carboxypeptidase A activity is 

about 6 times higher than carboxypetidase B. The total proteolytic activity as well as trypsin, 

chymotrypsin, aminopeptidase and carboxypeptidase activities did not differ during the 

vegetation season when WRC was actively feeding on the maize. The high activity of serine 

proteinases being the main target for many well known protease inhibitors seems to be 

promising way for the control of D. virgifera in future. 

This study was supported by the grant No. N N310 088936 from the Ministry of Science and 

Higher Education in Warsaw, Poland. 

- POSTERS - 75


P-15 

CHANGES OF CCAP INFLUENCE ON AGEING HEART 

OF TENEBRIO MOLITOR 

76 

Joanna Pacholska-Bogalska 1 , Kamila Woźniak 1 , Paweł Marciniak 1 , Monika 

Szymczak 1 , Grzegorz Rosiński 1 

1 Department of Animal Physiology and Development, Adam Mickiewicz University, 

Umultowska 89 Str., 61-614 Poznan, Poland, e-mail: pacholsk@amu.edu.pl 

Ageing of the organism is a natural biological process related to occurring 

adverse changes, which determine long life of the organism, at molecular, cellular and 

tissue level. Heart is one of the organs, which function during ageing is influenced by 

dysfunction such as arrhythmia, tachycardia or bradycardia. Physiological changes in 

adult heart of ageing insect are poorly known. There is no information about changes 

in the heart contractile activity parameters of T. molitor, its sensitivity to regulatory 

peptides influence or ion channels activity. 

One of the neuropeptides, which exerts cardiotropic activity, and can modulate 

the action of the insect heart is the crustacean cardioactive peptide (CCAP; 

PFCNAFTGCa). It has been identified in neurohormonal system of many insect 

species including the beetle T. molitor. The peptide has been shown to induce an 

increase in the frequency and amplitude of contraction of the T. molitor myocardium. 

The aim of the present study was the analysis of endogenic heart rhythm of the 

ageing T. molitor beetles and characterization of the CCAP influence on the ageing 

myocardium as well. 

We detected chronotropic and inotropic changes in endogenous rhythm of 

semi-isolated heart of ageing beetles. We also observed that stimulating impact of 

CCAP on the heart was age-dependant. CCAP application caused, depending on age, 

alternations in chronotropic and inotropic parameters of the heart contractions and the 

myocardium of the older beetles was less sensitive to the peptide. 

Grant support: N N303 805640 (National Science Centre) 

- POSTERS -


P-16 

HAEMOLYMPH PICTURE IN DESERT LOCUST AS 

AFFECTED BY TWO ENTOMOPATHOGENIC FUNGI 

BEAVERIA BASSIANA AND METARHIZIUM 

ANISOPLIAE VAR ACRIDIUM 

F. Bounaceur 1 , Z. Bissaad 2 , B. Doumandji-Mitiche 3 

1 Faculty of Science of nature and live, Ibn Khadoun University, Tiaret, Algeria, 

fbounaceur@yahoo.fr 

2 Department of Biology, Science faculty, University M’Hamed Bouguera, Boumerdes, 

Algeria, bissaad@yahoo.com 

3 Department of Agricultural and Forest Zoology, National School Agronomy, El- 

Harrach, Algiers, Algeria, doumandjimitiche@yahoo.fr 

Interest of using pathogens as biological control agents against locust and 

grasshoppers has grown since the last major locust plague of the 1980s. Millions liters 

of chemical pesticides were used to combat this plague, which led to an outcry among 

environmental groups. The most extensively studied pathogens are the 

entomopathogenic fungi such as M. anisopliae var. acridum and Beauveria bassiana 

(Greathead D.J. (1992) Proceeding of workshop held at the International Institute of 

Tropical Agriculture, Cotonou, 29April- 1 May 1991,105-121). The use of pathogens 

may offer an environmentally sound method for the management of grasshoppers and 

locusts, hyphomycete fungi are the most promising candidates (Perior C. and 

Greathead D.J. (1989) FAO Plant Protection Bulletin. Vol. 37, 37-48). 

The infection of desert locust of both fungi (sole or in combination) caused reduction 

of the total haemocytes count and resulted in changes of the percentages of the 

haemocyte types. The combined infection caused potential effect at all observations 

when 5 th nymphal instar was treated with 10 spores/nymph. While in case of both 

fungi (2x103 spores/nymph), there was an additive effect on day 7 and potential effect 

on days 1, 3, 6 and 9 post infection, the combined infection also accelerated the 

mortality. 

Keywords: Schistocerca gregaria, Metarhizium anisopliae var. acridum, Beauveria bassiana,, toxicity 

effect, haemolymph picture. 

- POSTERS - 77


P-17 

HAEMOLYMPH CHEMICAL COMPONENTS IN 

DESERT LOCUST AS AFFECTED BY TWO 

ENTOMOPATHOGENIC FUNGI BEAUVERIA BASSIANA 

AND METARHIZIUM ANISOPLIAE VAR ACRIDUM 

78 

F.Z. Bissaad 1 , F. Bounaceur 2 , B. Doumandji-Mitiche 3 

1 Department of Biololgy, Science faculty, University M’Hamed Bouguera, Boumerdes, 

Algeria, bissaad@yahoo.com 

2 Faculty of Science of nature and live, Ibn Khadoun University, Tiaret, Algeria, 

fbounaceur@yahoo.fr 

3 Department of Agricultural and Forest Zoology, National School Agronomy, El- 

Harrach, Algiers, Algeria, doumandjimitiche@yahoo.fr 

Locusts and grasshoppers are major economic pests of crops and grasslands 

throughout the world's dry zones, their attacks attract much public attention, few other 

pests make headline news (Lomer, C. J. et al. (1999) Agricultural and Forest 

Entomology.1:71-88). Desert locust of Schistocerca gregaria (Forskal, 1775) 

consumes approximately their own weight (ca. 2g) of fresh vegetation each day. 

Swarms often contain 50 million individuals per Km², so that even a moderate swarm 

measuring 10 Km² could consume about 1000 tons of fresh vegetation daily during 

migration (Coper, A. (1982) In Van Huis, A. (Ed) Proceeding of the seminar held in 

Wageningen. 11-17). 

The aim of the present study is to evaluate the susceptibility of S. gregaria to the two 

entomopathogens: Metarhizium anisopliae var. acridum (Metchnikoff) Soroken and 

Beauveria bassian a(Bals.) Vuill. 

In this study, the effects of M. anisopliae var. acridum, B.bassiana. sole and their 

combination were studied on the haemolymph chemical composition of the 5 th 

nymphal instar. The infection of both fungi (sole or in combination) caused reduction 

in chemical composition of haemolymph in the 5 th nymphal instar of the desert locust 

S. gregaria, also reduced the activity of Acid phosphatase, prophenyleoxidase, and 

phenyleoxidase. 

Keywords: Schistocerca gregaria, Metarhizium anisopliae var. acridum, Beauveria bassiana, toxicity 

effect, haemolymph. 

- POSTERS -


P-18 

CONTRIBUTION TO THE FOLLOW-UP OF THE 

DYNAMICS OF THE POPULATIONS OF TUTA 

ABSOLUTA MYRICK 1917 ON TOMATO CULTURE 

EXCEPT GROUND IN ALGERIA 

Chebouti-Meziou Nadjba 1 , Bousaad Friza 1 , Belhadj Samira 1 Et Doumandji 

Salahedine 2 

1. Departement of biology faculty of science University M hamed Bougara Boumerdes, 

Algeria 

2. Uuniversity National of Agronomy Algeria 

The mineuse one of the tomato Tuta absoluta Myrick 1917, is a species known 

as of forty. It was announced in Algeria in May 2008. It causes considerable damage, 

in particular on the vegetable farming of the family of solanaceous, they are ravageurs 

very frightening. She causes losses from 80 to 100 % on tomato culture. The present 

study consists in making a follow-up of the dynamics of the populations of Tuta 

absoluta on culture except ground (coconut fibre) of tomato in the area of hamiz 

(Algeria). An intense activity of Tuta absoluta was recorded the end May 2010 is 

successively 59, 49, 63, and 10 individuals for the second, third, larval fourth, stages 

and the chrysalis. It is to be announced a strong pullulation on the Pristyla variety 

compared to the variety will jawhara is a rate from 20 to 44 %. The small percentages 

of attack are noted on the tomato cherry (K6, Sontonio and Minilon) that is to say 15 

to 28 %. The mines dug on the sheets of the round varieties are more important than 

those dug on the sheets of the cherry varieties. 

Keywords: Tuta absoluta, tomato, damage, dynamics of the populations, Algeria. 

- POSTERS - 79


P-19 

REPELLENT ACTIVITY OF FLAVONOID AGLYCONES 

AND CATECHOL AGAINST RHOPALOSIPHUM PADI L. 

80 

G. Chrzanowski, P. Czerniewicz, S. Goławska, H. Sytykiewicz, 

B. Leszczyński and A. Kozak 


Sciences and Humanities, Siedlce, Poland, e-mail: grzegorzc@uph.edu.pl 

Flavonoids are a group of polyphenolic compounds widely distributed in plant 

kingdom. They are components of different plant organs, as inflorescences, leaves and 

fruits. Numerous positive effects of flavonoids have been described and their activities 

are mostly connected with antioxidant and free radical scavenging. Moreover, 

polyphenolic compounds are plant resistance factors towards insects and pathogens. 

Thus, the aim of the study was to examine influence of selected flavonoid aglycones 

and catechol on behaviour of wingless females and larvae of bird cherry-oat aphid 

(Rhopalosiphum padi L.). 

Catechol, (+)-catechin, (-)-epicatechin, quercetin, apigenin, myricetin and luteolin 

were obtained from Sigma Chemical Co. (St. Louis, Mo, USA). Effect of four 

concentrations (0.001 mM, 0.01 mM, 0.1 mM and 1 mM) of polyphenols on aphid 

settling behaviour was studied in choice-tests. 

Apigenin and myricetin were strong repellents towards apterous females and larvae of 

R. padi in all concentrations. Moreover, (+)-catechin and (-)-epicatechin showed a 

negative influence on settling behaviour of the bird cherry-oat aphid in higher 

concentrations, instead catechol and luteolin weakly effected aphid settling. 

This study was carried out with the support of the Ministry of Science and Higher Education of the 

Republic of Poland (Project No. N N310 298739). 

- POSTERS -


P-20 

PLANT EXTRACTS AS BIOPESTICIDES 

Michalczyk A., Cieniecka-Rosłonkiewicz A., Morytz B., Kazimierczak J. 

Institute of Organic Industry in Warsaw , Michalczyk@ipo.waw.pl 

The ecological trends in agriculture and problems of the new pesticides 

introduction to the market caused the natural compounds from plants became the 

subject of researcher is interest. Plants produce a very diverse range of secondary 

metabolites such as essential oils, tannins glycosides, organic acids, alkaloids, 

saponins. The newest literature sources revealed that extracts from the bark and 

needles of Taxus baccata as well as extracts from Cinnamomum cassia bark and the 

seeds of Myristica fragrans showed the activity against Tetachynus urticae, Blattela 

germanica, Tribolium castaneum, Musca domestica. The alcoholic extracts from 

cinnamon bark and from seeds of nutmeg showed also activity against fungal and 

bacterial plant patogents. The aim of this work was the extraction of bioactive 

compounds from above mentioned plants using organic solvents and 4 ionic liquids. 

The activity of extracts was evaluated against Musca domestica, Blatta orientalis, 

Tetrachynus urticae and plant pathogenic fungi and bacteria. Extracts were prepared 

using traditional organic solvents such as hexane and ethyl acetate and 4 ammonium 

ionic liquids. It was found that extracts from yew (Taxus baccata)obtained using ionic 

liquids exhibited the weak acaricidal and insecticidal action. Extracts from yew 

obtained by organic solvents didn‟t exhibit the activity against used bioindicators. 

Extracts from nutmeg and cinnamomum obtained using ionic liquids revealed the high 

insecticidal activity. The above mentioned extracts showed also the fungicidal and 

bactericidal action against plant pathogens. 

- POSTERS - 81

82 


P-21 

CHARACTERIZATION OF HEMOCYTES FROM THE 

MEALWORM BEETLES TENEBRIO MOLITOR AND 

THEIR RESPONSE TO NEB-COLLOOSTATIN 

Czarniewska E. 1 , Mrówczyńska L. 2 , Kuczer M. 3 , Rosiński G 1 . 

1 Department of Animal Physiology and Development, A. Mickiewicz University, 

Poznań, Poland, czarniew@amu.edu.pl 

2 Department of Cell Biology, A. Mickiewicz University, Poznań, 

3 Institute of Chemistry, Wroclaw University, Wroclaw, Poland 

In this study, we demonstrate that the hemolymph from T. molitor adults 

contains three hemocyte types: prohemocytes, plasmatocytes, and granulocytes, 

distinguished by light and scanning electron microscopes. Plasmatocytes and 

granulocytes, involved in cellular defense reaction, are the most abundant cells type of 

the hemolymph. Prohemocytes, multipotent stem cells, comprised less than 8 % of the 

total hemocytes population. 

Hemocytes were exposed in vivo to Neb-colloostatin, gonadotropic peptide isolated 

from the ovaries of the flesh fly Neobellieria bullata, by its injecting into the body 

cavity. The obtained results indicate that injection with Neb-colloostatin affected the 

morphology and viability of hemocytes in the dose-dependent manner. Utilizing 

different methods, the typical features of apoptosis in every type of Neb-colloostatintreated 

hemocytes were shown, i.e. induction of apoptotic markers (the caspases), 

DNA fragmentation, and membrane blebbing. Authophagic vacuoles 

(authophagosomes) were detected in none of Neb-colloostatin-treated hemocytes 

types. The F-actin cytoskeleton has been found aggregated into distinctive patches 

after peptide treatment. 

Neb-colloostatin did not impair granulocytes and plasmatocytes phagocytosis of 

foreign material, including the latex beads, both in vitro and in vivo. 

This is the first report that Neb-colloostatin exposure induces apoptosis and the 

cytoskeleton disruption in insect hemocytes. 

Acknownledgements: This work was supported by a grant from Ministry of Science and Higher 

Education N N 204085638 

- POSTERS -


P-22 

Neb-COLLOOSTATIN INDUCES FOLLICULAR 

ATRESIA DURING TENEBRIO MOLITOR OOGENESIS 

Czarniewska E. 1 , Mrówczyńska L. 2 , Kuczer M. 3 , Rosiński G 1 . 

1 Department of Animal Physiology and Development, A. Mickiewicz University, 

Poznań, Poland, 2 Department of Cell Biology, A. Mickiewicz University, Poznań, 3 

Institute of Chemistry, Wroclaw University, Wroclaw, Poland 

czarniew@amu.edu.pl 

Our earlier physiological studies showed that Neb-colloostatin 

(SIVPLGLPVPIGPIVVGPR), peptide with the oostatic properties isolated from the 

ovaries of the flesh fly Neobellieria bullata, exerts gonadoinhibitory effect, reduces the 

number of eggs and their hatchability, and delays the embryonic development in T. 

molitor. [1] 

In this study, we report that the injection of 2 or 20 ng of Neb-colloostatin into the 

body cavity induces an atresia of both previtellogenic and vitellogenic follicles in the 

ovaries of the 4-day-old females of T. molitor. We used several cytochemical methods 

to demonstrate the morphology of the ovaries of Neb-colloostatin treated females. The 

analysis in light and confocal microscopes showed that the peptide, in both doses, 

induces the atresia in the oocyte and in the follicular cells. In atretic follicular cells, 

we observed typical features of apoptosis: (1) changes in the chromatin organization, 

(2) induction of the caspases activity and (3) disorganization of the actin cytoskeleton. 

In addition autophagic vacuoles were present in the follicular cells. 

This is the first report that Neb-colloostatin induces apoptosis in insect ovariole. We 

present data showing that Neb-colloostatin exerts pleiotropic activities in regulation of 

insect oogenesis. 

[1] Wasielewski O., Rosiński G. (2007) Arch. Insect Biochem. Physiol. 64, 131-141. 

Acknownledgements: This work was supported by a grant from Ministry of Science and Higher 

Education N N 204085638 

- POSTERS - 83

84 


P-23 

EFFECT OF PHENOLIC ACID EXTRACT FROM 

JUGLANS REGIA L. ON BIRD CHERRY-OAT APHID 

P. Czerniewicz, G. Chrzanowski, I. Sprawka, H. Sytykiewicz and B. Leszczyński 


Sciences and Humanities, Siedlce, Poland, e-mail: pawelc@uph.edu.pl 

Aphids are important pests of cereals that can reduce cereal yield and quality 

due to its capacity for extremely rapid population growth. Frequent application of 

synthetic insecticides is associated with reduction quality of grains, negative impact on 

environment and increased insect resistance. An alternative to such compounds may be 

extracts containing plant allelochemicals. One of the most active anti-herbivore 

chemicals produced by plants are phenolics. The aim of this work was to examine the 

effect of phenolic acid extract from leaves of walnut on bird cherry-oat aphid 

(Rhopalosiphum padi L.) settling behaviour and development. 

Phenolic acid fraction from mature walnut leaves was extracted and evaporated to 

dryness. The obtained residue was used to prepare working solutions for bioassays. 

Aphid settling was assessed using choice-test, where aphids were exposed to four 

concentrations of phenolic acids: 0.01%, 0.025%, 0.05% and 0.1%. Effect of the 

phenolic acids on the antibiosis parameters was determined after application 0.1% and 

0.2% solutions on R. padi individuals reared on winter wheat seedlings. 

The obtained results revealed that phenolic acid extract from walnut negatively 

influenced the bird cherry-oat aphid settling behaviour and development. Wheat 

seedlings treated with higher concentrations of phenolic acids reduced aphid settling 

during the entire time of the experiment. It was also shown that application of walnut 

extract prolonged the aphid prereproductive period, decreased its fecundity and 

increased mortality compared to the aphids reared on control plants. Aphids exposed to 

walnut extracts also showed smaller values of intrinsic rate of natural increase (rm). 

The lowest values of rm and daily fecundity were characteristic for the insects treated 

with 0.2% solutions. In this case the analysed antibiosis parameters were reduced 

about 30%. The obtained results suggest that phenolic acid extracts from walnut can be 

used as a source of natural biopesticides for cereal aphids. 

This study was carried out with the support of the Ministry of Science and Higher Education of the 

Republic of Poland (Project No. N N310 298739). 

- POSTERS -


P-24 

HALOLACTONES WITH THE p-MENTHANE SYSTEM – 

ACTIVE ANTIFEEDANTS AGAINST APHID MYZUS 

PERSICAE 

A. Grudniewska 1 , K. Dancewicz 2 , B. Gabryś 2 , A. Gliszczyńska 1 , Cz. Wawrzeńczyk 1 

1 Department of Chemistry, Wrocław University of Environmental and Life Sciences, 

Wrocław, Poland, e-mail: czeslaw.wawrzenczyk@up.wroc.pl 

2 Department of Biology and Ecology, University of Zielona Góra, Zielona Góra, 

Poland, e-mail: b.gabrys@wnb.uz.zgora.pl 

Lactones with the p-menthane system are are known for a broad variety 

of biological activity (Gaudin, J. M. (2000) Tetrahedron 56, 4769; Wawrzeńczyk, C. et 

al. (2005) Pol. J. Environ. Stud. 14, Suppl. II, 69). Our interest in the synthesis of this 

group of compounds is connected with their feeding deterent activity against insects. 

Here we present the results of antifeedant activity of five racemic and five 

enantiomerically enriched (ee = 91-98%) halolactones (1-5) with the p-menthane 

system against Myzus persicae. The lactones were obtained in the stereoselective 

syntheses from racemic monoterpene ketone – piperitone (6) (Grudniewska A. et al. 

(2011) Org. Biomol. Chem. Submitted). 

All of the iodo-, bromo- and chlorolactones showed high antifeedant activity 

against peach-potato aphid, which was in contrast to starting piperitone that had weak 

attractant properties. The results showed that the introduction of the lactone moiety 

and halogen atom into the p-menthane skeleton produced feeding deterrent activity of 

the compounds. 

- POSTERS - 85

86 


P-25 

EFFECTS OF DIFFERENT COMPOUNDS ON 

ALIMENTARY CANNEL ALPHA-AMYLASE OF 

MEDITERRANEAN MEAL MOTH 

Farshbaf Pourabad, R. 1 , Dastour, L. 2 , Ebrahimzade, M. 3 , Mohammadi, D. 4 




lailadastoor@yahoo.com 


m.ebrahimzadeh88@yahoo.com 



Mediterranean meal moth is one of the most important stored products pests 

that impose damages to storage products such as meals. In this study effects of 

different concentrations of some compounds such a CaCl2, MgCl2, Ascorbic acid and 

Ammonium sulfate on alimentary alpha-amylase of above mentioned insect were 

investigated. Studied insects were reared on general wheat meals on controlled 

condition and the midgets of 5 th larval stages were used for enzyme assays. Enzyme 

activity were measured using specific diagnostic kit (Pars-Azemon, Iran) using an auto 

analyzer system. The results revealed that CaCl2 and MgCl2 in male larvae increased 

enzyme activity dramatically but in females had a minor effect. The effects of ascorbic 

acid was dose and gender dependent, with increasing the dose to 3 mM enzyme 

activity in male insects increased but 4 and 5 mM caused a sharp decrease in enzyme 

activity and about female insects increasing the dose had inverse effect on enzyme 

activity. Ammonium sulfate inhibitory activity in females was more than males and in 

both sexes with increasing the concentrations enzyme activity decreased. 

- POSTERS -


P-26 

BIOLOGICAL PARAMETERS OF RHOPALOSIPHUM 

MAIDIS (FITCH) (HOMOPTERA: APHIDIDAE) ON 

CERTAIN CORN CULTIVARS 

Erol Bayhan*, Selime Ölmez Bayhan* 

*Department of Plant Protection, Agricultural Faculty, Dicle University, Diyarbakır- 

Turkey. 

E-mail: erolbayhan@gmail.com 

The development, survival rate, reproduction and biological parameters of the 

corn leaf aphid, Rhopalosiphum maidis (Fitch), on five corn cultivars were evaluated 

at 25°C under laboratory conditions. The corn leaf aphid reared on Gözdem had the 

highest fecundity (69.90 offspring/aphid). The lower percentage survival rates were 

observed on 359P12 corn cultivar than that of the others. Net reproductive rate (Ro) 

was lowest (66.85 offspring/aphid/generation) on 359P12 corn cultivar. The intrinsic 

rate of increase (rm) was lower on 359P12 corn cultivar than that of the other corn 

cultivars. 

Keywords: Rhopalosiphum maidis, biological parameters, corn cultivars. 

- POSTERS - 87

88 


P-27 

HOST AGE PREFERENCE BY DIAERETIELLA RAPAE 

(M’INTOSH) (HYMENOPTERA: BRACONIDAE: 

APHIDIINAE) ON CABBAGE APHID, BREVICORYNE 

BRASSICAE (L.) (HEMIPTERA: APHIDIDAE) 

Selime Ölmez Bayhan 1 , Mehmet Rifat Ulusoy 2 , Erol Bayhan 1 

1 Dicle University, Agricultural Faculty, Plant Protection Department, 

Diyarbakır-Turkey 

2 Cukurova University, Agricultural Faculty, Plant Protection Department, 

Adana-Turkey 

The instars preference of Brevicoryne brassicae by the parasitoid Diaeretiella 

rapae was investigated at 20 1 0 C, 65 5 % r.h. and 16 L: 8 D photoperiod. In the 

first trial, mixture of all stages of cabbage aphid in equal ratios, and in second trial, 

separately for each stage was used as host for parasitisation. The objective was to 

identify the most preferred age of cabbage aphid for parasitism by D. rapae. 

As a result of the study, both mixed with cabbage aphid of different biological 

stages and given separately, the aphid of the second and third periods were preferred 

by D. rapae. Both at trial as a result of statistical analysis performed between the 2nd 

and 3rd nymphal stages despite the lack of significant quantitative difference between 

them was found to be more interference in the 2nd instar. 

- POSTERS -


P-28 

ALFALFA LUTEOLIN, TRICIN AND CHRYSOERIOL 

GLYCOSIDES AND THEIR EFFECT ON THE PEA 

APHID 

S. Goławska 1 , G. Chrzanowski 1 , I. Łukasik 1 , I. Sprawka 1 , I. Kapusta 2 , B. Janda 2 

1 Department of Biochemistry and Molecular Biology, Siedlce University of Natural 

Sciences and Humanities, Siedlce, Poland, e-mail: sylwia@uph.edu.pl 

2 Deprtment of Biochemistry, Institute of Soil Science and Plant Cultivation, Puławy, 

Poland 

Alfalfa (Medicago sativa L., Fabaceae) is an important agricultural and 

commercial crop. Unfortunately, these economically important crops have serious 

pest, the pea aphid, Acyrthosiphon pisum Harris (Homoptera: Aphididae). Beside the 

nutritional components alfalfa also produce a variety of secondary metabolites. Many 

of them help to protect the plant against herbivores. It has been shown that plant 

secondary metabolites can seriously affect the insects behaviour, physiology and 

metabolism and as a result reduce their populations on resistant crops. Among the 

secondary metabolites of alfalfa, that have been identified and quantified flavonoids 

are suggested to play an important role in defense against herbivores, but little 

information on such effect of the alfalfa flavonoids exist. Due to such deleterious 

effects of flavonoids on insect species, the aim of the study was identification, 

quantification and determination of an influence of the flavonoids content on the pea 

aphid. 

Liquid chromatography (HPLC) was used to determine the flavonoid profiles, 

especially luteolin, tricin and chrysoeriol glycosides, their total concentration, as well 

as changes in the amount of eight flavones found in the aerial parts of alfalfa 

(Medicago sativa L.) (Fabaceae) Radius cv. for three vegetative stages, uninfested and 

infested by the pea aphid (Acyrthosiphon pisum Harris) (Homoptera: Aphididae). It 

was shown that both control and infested green aerial parts of alfalfa plants had similar 

flavonoid profiles. The dominant flavonoid of alfalfa was compound 7-O-[2-Oferuloyl- 

-D-glucurono-pyranosyl(1 2)-O- -D-glucuronopyranosyl]-4‟-O- -Dglucuronopyranosideluteolin. 

Compound 7-O-{2‟-O-feruloyl-[ -Dglucuronopyranosyl(1 

3)]-O- -D-glucuronopyranosyl(1 2)-O- -D- 

glucuronopyranosidetricin was present in the smallest amounts. The total 

concentration of flavones was rather high and ranged from 9.68 to 11.89 mg/ g d.m., 

but there were no significant differences between uninfested and infested alfalfa 

plants. Pea aphid abundance on the alfalfa was significantly related to the 

concentration of studied flavonoid glycosides. This finding may indicate the 

importance of flavonoid glycoside forms in the alfalfa resistant towards the pea aphid. 

- POSTERS - 89


P-29 

EFFECTS OF DIFFERENT COMPOUNDS ON 

DIGESTIVE LIPASE ACTIVITY OF GREAT WAX 

MOTH GALLERIA MELLONELLA L. (LEP., PYRALIDAE) 

90 

Mohammadi D. 1 , Farshbaf Pourabad R. 2 , Alipour, E. 3 , Valizade M. 4 






ent20032003@yahoo.com 

4 Department of agronomy and Plant breeding, University of Tabriz, Tabriz, Iran, 

mvalizadeh@tabrizu.ac.ir 

Lipases, able to catalyze the hydrolysis of oils/fats as well as a wide variety of 

reactions in aqueous and nonaqueous phase. The great wax moth, Galleria mellonella 

feeds on wax and the lipase are one on the most important enzymes in alimentary 

cannel of this insect. For inhibitory or activity assays, GWM were reared on wax based 

artificial diet in controlled condition. After dissection alimentary cannel of last larval 

instars were used as lipase sources. The enzyme activity was measured using specific 

diagnostic kit using an auto analyzer system. Effects of different concentrations of 

some compounds on lipase activity were studied. The results revealed that lipase 

activity decreased in the presence of 75% ethanol and 5mM magnesium nitrate in 

addition with different concentrations of SDS (1-5 mM). Potassium phosphate and 

NaCl increased the enzyme activity especially in 5mM concentrations. Other tested 

compounds such as different concentrations of EDTA had no effects on enzyme 

activity. In overall the applied compounds in biochemistry and molecular biology such 

those ones were tested in this study could have diverse effects on enzyme activity; we 

can look at them as a compounds for changes in enzyme activity in biochemistry 

assays. 

- POSTERS -


P-30 

POPULATION DYNAMICS OF THE INVASIVE PEST 

SPECIES DIABROTICA V.VIGIFERA IN SOUTHERN 

EUROPE 

Hans E. Hummel 1,2 and M. Bertossa 3 

1 Chair of Organic Agriculture, Justus-Liebig-University Giessen, Germany. 

Hans.E.Hummel@agrar.uni-giessen.de 

2 Illinois Natural History Survey, Prairie Research Institute, University of Illinois, 

USA. 

3 Agroscope-Changings-Wädenswil ACW, Centro di Cadenazzo, CH-6594 Contone, 

Switzerland. Mario.bertossa@acw.admin.ch 

Diabrotica v. virgifera Le Conte (Coleoptera: Chrysomelidae), the western 

corn rootworm (WCR) is an invasive species to Europe. It first arrived in Europe from 

its native territory in the Americas about two decades ago. After multiple introductions 

by airplanes it is spreading from a number of foci at a considerable rate and is 

establishing itself firmly and irreversibly. We studied its population dynamics in 

Hungary, Serbia, Croatia, Romania, Slovenia, Southern Germany, and most 

extensively in Southern Switzerland. 

In the Canton Ticino, a minute WCR population was first detected in 2000 and 

dramatically increased after the year 2000. In response, Swiss authorities, disregarding 

common insecticide usage trends, decided to take recourse solely to the traditional 

management system of area-wide crop rotation. From the year 2003 onward 

mandatory crop rotation has been enforced and resulted in a stabilization of WCR at 

population levels below the economic threshold. No further increase of populations 

has been observed, in spite of annual northbound flights of WCR from the Lombardy 

region of Northern Italy reaching up to the foothills of the Gotthard Mountains in the 

central chain of the Alps. 

Our methods simply consisted in constantly monitoring, on a yearly basis, the beetle 

flights with sensitive sex pheromone and kairomone baited sticky traps (Bertossa & 

Hummel 2008, Hummel et al. 2009). This inexpensive tool allowed the establishment 

of flight curves in time and space. So, under marginal expenses, area-wide pest 

management can be practiced exclusively with cultural techniques. This approach can 

be recommended for adoption by other European countries with suitable geographical 

features. 

In addition, our studies indicate that WCR beetles are spreading along preferred flight 

paths and can be carried by wind currents over long distances. Maize is not a 

mandatory crop. The larvae may survive on alternative hosts and be trapped at nonmaize 

sites which confirms earlier observations of the senior author in Illinois. 

Bertossa, M. & Hummel, H.E. (2008) Comm. Appl. Biol. Sci., Ghent Univ. 73, 421. 

Hummel et al. (2009) Proc. DPG-BCPC Berlin. P 338. 

- POSTERS - 91


P-31 

ORGANIC ELECTROSPUN NANOFIBERS AS NOVEL 

DISPENSERS FOR INSECT PHEROMONES: AN ENTIRELY 

NOVEL TECHNICAL APPROACH TO MATING DISRUPTION IN 

IPM PROGRAMS 

92 

Hans E. Hummel 1,2 , D.F. Hein 1 , I. Lindner 1 , G. Leithold 1 , A. Greiner 3 , 

C. Hellmann 3 , J.H. Wendorff 3 , M. Breuer 4 , A. Kratt 5 , H. Kleeberg 5 

1 Justus-Liebig-University, Organic Agriculture, Karl-Gloeckner-Str.21c, D-35394 

Giessen, Germany; hans.e.hummel@agrar.uni-giessen.de 


Champaign, Illinois 61820, USA 

3 Philipps-University Marburg, Institute of Macromolecular Chemistry, Hans- 

Meerwein-Str., D-35032 Marburg, Germany 

4 State Institute of Viticulture and Enology Freiburg, Department of Ecology, 

Merzhauserstr.119, D-79100 Freiburg, Germany 

5 Trifolio-M GmbH, Dr.-Hans-Wilhelmi-Weg 1, D-35633 Lahnau, Germany; 

Armin.Kratt@trifolio-m.de 

Conventional pheromone dispensers have a long history of development. In 

spite of nearly forty years of improvements they still fall short of the ideal in 

mechanical and UV stability, predictability of release rate, long lasting efficacy, cost, 

and environmental concerns over limited biodegradability of residual polymer 

material, to name a few obstacles. With modern requirements such as sustainability as 

well as mechanical pheromone distribution in mind, we searched for practical ways of 

overcoming most of past shortcomings. We now have data which prove successful 

dispensing of sex pheromones by nanofibers with the aim of IPM by behavioral, 

nontoxic intervention against mating activities of the European grape vine moth 

Lobesia botrana (Lepidoptera:Tortricidae), a severe pest in vineyards. At Freiburg, 

Germany, electrospun, biocompatible fibers of organic nanomaterial served as 

dispensers during the growing season of 2009 and 2010. - According to Greiner & 

Wendorff (2007), such electrospun nanofibers offer new territory with numerous 

applications. In our hands, they are a viable alternative to currently available 

dispensers. A fine non-woven fleece of Ecoflex ® nanofibers charged with (E,Z)-7,9dodecadien-l-ol 

acetate was deposited in a vineyard. According to the standard 

procedure of Doye (2006), walk-in field cages with reared male and female L .botrana 

moths served as test sites for quantifying the disruption success over time. Three 

consecutive replications were run during 2009 with segments of electrospun 

nanofibers, followed by one experiment with a continuous sheets of nanofibers 

exposed on a mechanical carrier. No treatment (blank) and commercial Isonet ® 

dispensers served as negative and positive controls, resp. Disruption effects observed 

were statistically significant for 3 weeks. With modifications employed in 2010, these 

effects could be extended up to seven weeks (Hummel et al. 2010, Hein et al. 2011). - 

Application of this novel approach to other commercially important pest insects is 

imminent, as are steps taken toward mechanization of the procedure. 

Doye, E. (2006) PhD-Thesis. TU Kaiserslautern. Department of Biology. 

Greiner, A. & Wendorff, J.H. (2007) Angew. Chem. Int. Ed. 46, 5670. 

Hein et al. (2011) Beiträge zur 11. Wissenschaftstagung Ökologischer Landbau, pp 318. 

Hummel et al. (2010) In: Research Journal RMUTT Special Issue 14: D1-6, Thailand. 

- POSTERS -


P-32 

HALF A CENTURY OF PHEROMONE SCIENCE 

Hans E. Hummel 1, 2 , E. Hecker 3 , K.-E. Kaissling 4 

1 Justus-Liebig-University, Organic Agriculture, Karl-Gloeckner-Strasse 21C, 

D-35394 Giessen, Germany. Hans.E.Hummel@agrar.uni-giessen.de 


Champaign, Ill. 61820, USA 

3 Deutsches Krebsforschungszentrum, Im Neuenheimer Feld, D-69120 Heidelberg , 

Germany 

4 Max-Planck-Institut für Ornithologie, D-82346 Seewiesen-Andechs, Germany 

Contemporary biologists take the term as well as the concept of "pheromones" 

for granted. Yet, it was a sensational and controversial event when the January 1959 

issue of Nature magazine published a two page communication by Karlson and 

Lüscher on "Pheromones, a new term for a class of biologically active substances". 

This seminal paper appeared simultaneously with the first identification of the 

chemical structure of bombykol by Butenandt, Hecker and coworkers (1959). Bombyx 

mori, the silkworm moth, with its main pheromone component (E,Z)-10,12hexadecadien-l-ol 

served as a prototype for an entirely new class of exogenously 

active natural signal compounds. Two years later, the total synthesis via several 

independent routes by Butenandt, Hecker et al. 1961 concluded 20 years of pioneering 

work into unknown territory. Immediately, the impact of these discoveries for chemical 

communication, chemical ecology, sensory physiology and practical plant protection 

was recognized. Butenandt‟s forecast of 1939 turned out to be viable. 

Nothing documents the significance of pheromones better than close to twenty 

thousand original research articles and more than forty monographs and major reviews 

which appeared in the literature up to know, including a review of 1939 – 1961 by 

Hecker and Butenandt (1984). Today, pheromones are known from pest insects in all 

major food and fiber crops and from stored products. Most prominent are insect sex 

attractants. But chemical signals for territorial marking, trail following, general 

chemical orientation to host plants and oviposition sites, hive cohesion and caste 

determination in social insects are also known. In insect pest management, 

pheromones and related kairomones play a key role as lures in traps for monitoring, 

mass trapping and for male disorientation/mating disruption. In favorable cases, 

pheromones can compete with non sustainable pesticides which provoke non-target 

toxicity, general ecotoxicity and resistance. Today, pheromones of other arthropods 

including spiders are also known. - Progress in vertebrate pheromone research is 

noteworthy, but slower. If exploited, its long term impact on animals and human 

society, however, may be enormous. - Pheromone research influenced other fields like 

analytics at the nanogram scale, electrophysiological methods including single cell 

recording, and biochemical genetics. Without pheromones, entomology would be like 

a meal without spices. 

Butenandt A (1939) In: Jahrbuch der Preußischen Akademie der Wissenschaften. P 97. 

Butenandt A. et al. (1959) Z. Naturforschung. 14, 283. 

Butenandt A, Hecker E (1961) Ang. Chem. 73, 349. 

Hecker E, Butenandt A. (1984) In: Techniques in pheromone research. Springer Verlag. P 1. 

Karlson P, Lüscher M (1959) Nature. 183, 55. 

- POSTERS - 93


P-33 

DOES ALPHA-CYPERMETHRIN HAVE INFLUENCE ON 

BODY MASS OF THE NON-TARGET PREDACIOUS 

CARABID BEETLE? 

94 

I. Kivimägi, A. Ploomi, L. Metspalu, E. Ńvilponis, K. Jõgar, K. Hiiesaar, E. 

Veromann, M. Mänd, A. Kuusik 

Department of Plant Protection, Estonian University of Life Sciences, Tartu, Estonia; 

e-mail: irja.kivimagi@emu.ee 

Carabid beetles (Coleoptera: Carabidae) are species rich group of important 

natural pest-control agents in agricultural fields. The aim of this study was to test the 

effect of widely used synthetic pyrethroid insecticide alpha-cypermethrin on 

predacious non-target carabid beetle (Platynus assimilis Payk. 1790) under laboratory 

conditions. The pyrethroid is a neuropoison acting on the axons in the peripheral and 

central nervous system. The adult beetles (n = 60) were divided into two groups: 

control and test. Body mass of the beetles was measured before the treatment and in 

the end of the study (14 th day). Beetles were treated with Fastac 50, commercial 

formulation of alpha-cypermethrin (a.i. 50g l -1 ), by dose 0.15%, at recommended 

agricultural application rate. Food pieces were dipped for 10 seconds into the distilled 

water (control) or treatment (test). The beetles were kept 14 days in plastic boxes (0.5 

l) individually on moistened tissue and with free access to manipulated food, at room 

temperature. The measured body masses increased 4 mg in control group, but no 

increase in treated group was detected. According to our results, treatment by alphacypermethrin 

affects insects feeding habits or probably inhibits or disrupts energy 

production from food, which also could reduce predation in agroecosystems. 

Keywords: alpha-cypermethrin, body mass, carabid beetle, non-target 

- POSTERS -


P-34 

BIOCHEMICAL CHARACTERIZATION 

OF HAEMOLYMPH AND DIGESTIVE SYSTEM 

PROTEASES OF NARANGA AENESCENS 

MOORE (LEP.: NOCTUIDAE) 

Ameneh Asadi 1 , Mohammad Ghadamyari 1 , Reza Hassan Sajedi 2 ,Jalal Jalali 1 

1- Department of Plant protection, Faculty of Agriculture, University of Guilan, Rasht, 

Iran 

2- Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran 

e-mail: mghadamyari@gmail.com 

Naranga aenescens L. (Lep.: Noctuidae), commonly known as the rice green 

caterpillar, is one of the most damaging pests of the rice crop in Iran. The biochemical 

properties of proteases from digestive system of N. aenescense were determined in this 

study. 5 th instar larvae were collected from rice seedling Oryza sativa L. „variety of 

Hashemi‟ in the northern provinces of Iran. 5 th instar larvae were randomly selected 

for measuring of enzyme activity. The effect of pH (4-11), temperature (15-75°C) and 

ions (5, 10 mM) and inhibitors (1, 5 mM) on digestive protease activities were 

measured using 50mM phosphate-glycine-acetate sodium buffer with azocasein as 

substrate. Tryptic and chymotrptic activities were assayed using BApNA and BTEE as 

substrates, respectively. Results showed that the optimal pH for protease in digestive 

system and haemolymph were 9 and 7, respectively. Also optimal temperature for 

protease activity in digestive system and haemolymph were 55°C and 35°C, 

respectively. Effect of ions on protease activity showed that Hg 2+ , Co 2+ , Fe 2+ , Zn +2 , 

Mn 2+ and Ca 2+ decreased protease activity in the digestive system of N. aenescense. 

Effect of inhibitors on protease activity in alimentary canal showed that iodoacetamide 

(5 mM), iodoacetate (5 mM), EDTA (2 mM), TLCK(1 mM), TPCK(1 mM) and PMSF 

(5 mM) decreased 39.52%, 54.46%, 28.3%, 19.16%, 33.12% and 24.77 % enzyme 

activity, respectively. Effect of inhibitors on protease activity in haemolymph showed 

that iodoacetamide (5 mM), iodoacetate (5 mM), EDTA (2 mM), TLCK(1 mM), 

TPCK(1 mM) and PMSF (5 mM) decreased 26.24%, 25.14%, 36.02%, 13.17%, 

24.45% and 17.36 % enzyme activity, respectively .The activity of total protease in 

alimentary canal and haemolymph were 3.345 and 0.446 OD/min/mg protein. Trypsin 

and chymotrypsin activities in digestive system wer 0.745 and 116.97 µmol/min/mg 

protein, respectively. 

- POSTERS - 95

96 


P-35 

BIOCHEMICAL CHARACTERIZATION OF α- AND β- 

GALACTOSIDASES FROM ALIMENTARY CANAL, 

SALIVARY GLANDS AND HAEMOLYMPH OF THE 

RICE GREEN CATERPILLAR, AENESCENS MOORE 

(LEP.: NOCTUIDAE) 

Ameneh Asadi 1 , Mohammad Ghadamyari 1 , Reza Hassan Sajedi 2 , Jalal Jalali 1 

1. Department of Plant protection, Faculty of Agriculture, University of Guilan, 

Rasht, Iran 

2. Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran. 

e-mail: mghadamyari@gmail.com 

Naranga aenescens L. (Lep.: Noctuidae), commonly known as the rice green 

caterpillar, is one of the most damaging pests of the rice crop in Iran. α- and βgalactosidases 

have an important role in the final stages of carbohydrate digestion in 

insect such as N. aenescens. 5 th instar larvae were collected from rice seedling Oryza 

sativa L. „variety of Hashemi‟ in the northern provinces of Iran. Larvae were randomly 

selected for measuring of enzyme activity. Alimentary canal, salivary gland and 

haemolymph of 5 th instar larvae were removed for analysis. The effect of pH (4-11), 

temperature (15-75°C) and ions on α- and β-galactosidase activities was measured 

using 40mM citrate-phosphate-acetate sodium buffer. The activity of α- galactosidases 

in the alimentary canal, salivary gland and haemolymph of 5 th instar larvae were 

1.12±0.01, 0.554±0.09 µmol/min/mg protein and 0.236±0.004 nmol.min/µl, 

respectively and the activity of β-galactosidases in the alimentary canal, salivary gland 

and haemolymph of 5 th instar larvae were 3.19±0.054, 0.623±0.034 µmol/min/mg 

protein and 0.633±0.074 nmol/min/µl, respectively. As calculated from Lineweaver- 

Burk plots, the Km value for α- galactosidases in alimentary canal and salivary gland 

were 2.63 and 3.11 mM, respectively. Also, the Km value for ß- galactosidases in 

alimentary canal and salivary gland were as 0.77 and 0.41 mM, respectively. 

Zymogram pattern in the native gel revealed that N. aenescense β- galactosidases in 

alimentary canal, salivary gland and haemolymph, had one isoform. Results showed 

that the optimal pH for α- galactosidases in alimentary canal, salivary glands and 

haemolymph were 6, 5 and 4, respectively. Optimal pH for β- galactosidases in 

alimentary canal, salivary glands and haemolymph were 6, 5 and 5, respectively. Also 

optimal temperature for α- galactosidases activity in alimentary canal, salivary gland 

and haemolymph were 55°C. Optimal temperature for β- galactosidases activity in 

alimentary canal was 45°C and salivary gland and haemolymph were 55°C. Effect of 

ions on the enzyme activity showed that NaCl, KCl, SDS, Urea and EDTA decreased 

even completely in all three tissue of N. aenescense α- and β-galactosidase. CaCl2 

decreased β-galactosidase activity from alimentary canal. 

Keywords: Rice green caterpillar; α- galactosidase; β-galactosidase; alimentary canal; Salivary glands; 

Haemolymph 

- POSTERS -


P-36 

COMPARISON OF MORALITY EFFECT BETWEEN 

NANO PESTICIDES AND PESTICIDES 

ON AGONOSCENA PISTACIAE UNDER LABORATORY 

CONDITIONS 

M. Rouhani and M. A. Samih 

Department of Plant Protection, Vali-e-Asr University, Rafsanjan, Iran 

Rouhani_valiasr@yahoo.com 

The pistachio is one of the most important horticultural products in Iran, 

Armenia, Iraq, Turkmenia and Turkey and famous as green gold tree. The common 

pistachio psylla, Agonoscena pistaciae Burckhardt and Lauterer (Hem.: Aphelaridae) 

is a key pest of pistachio trees throughout the pistachio-producing regions of Iran. The 

control of this pest has been based on insecticides with no attention to the pest density 

and economic damage. Nanotechnology is a form of molecular engineering, has been 

heralded as the „new technological revolution‟. Organic nanoparticles are the building 

blocks in various nanotechnology applications. In this research, we synthesis nanoamitraz 

and nano-thiametoxam, then was used bioassay test for comparison of 

determinate toxicity them with amitraz and thiametoxam. Considering that the main 

damage caused by the pest occurs in nymphs stages, the effect of nano-Amitraz, nanothiametoxam, 

amitraz and thiametoxam was estimated under laboratory condition 

(26±2ºC, 60-70 relative humidity and a photoperiod of 16L: 8D h) on the 5th instar 

nymphs of common pistachio psylla. The analysis of data was performed by SPSS 

software and the comparison of means by Dunkan test. The results showed that nano- 

Amitraz at 1.5µl/ml, nano-Amitraz at 1µl/ml, nano-thiametoxam at 300ppm, nanothiametoxam 

at 270ppm and Amitraz at 1.5µl/ml had the most mortality effect 

respectively. The results indicated that the nano-pesticide had a stronger reducing 

effect on the pistachio psylla nymphs than that of pesticide too. 

Keywords: Agonoscena pistaciae, bioassay, nano-pesticide, pesticide. 

- POSTERS - 97

98 


P-37 

ROLE OF BACULOVIRUSES AND BACILLUS 

THURINGIENSIS BACTERIA IN FOREST PEST 

REGULATION IN LATVIA 

Z. Metla 1 , L. Jankevica 1 , J. Halimona 1 , R. Seskena 1 , I. Zarins 1 

1 Laboratory of Experimental entomology, Institute of biology, University of Latvia, 

Salaspils, Latvia, e-mail: zmetla@inbox.lv 

Institute of Biology has been working on problems, which included development of 

microbiological methods for plant protection in Latvia for many years. Pest outbreaks 

cause significant losses for forestries during recent years. Chemical pest control 

becomes less and less acceptable. Considering the fact, that insecticides may reach 

food and drinking water, EU had established restrictions for applying chemical control. 

The aim of studies was to extend the knowledge of nucleopolyhedroviruses and 

Bacillus thuringiensis bacteria and to clarify their role in regulation of pest 

populations. 

Observations of natural epizootics of dendrophagous pests have been done regularly 

in different regions of Latvia: Venta, Padure, Ance, Engure, Kalsnava and Valka. 

Dead, infected and living insects were collected from natural habitats applying 

standard methods. After collecting living insects were placed in sterile isolators for 

observations. Reasons of death were evaluated by applying microscopy (Olympus CX 

41). The slosest attention was paid to the following collected insect species: 

Neodiprion sertifer, Bupalus piniarius,Diprion pini. 

Nucleopolyhedroviruses were isolated from 15 forest pest species. Granuloviruses 

were isolated from B. piniarius. 

During the study nucleopolyhedrovirus isolates were obtained, which had high 

virulence and may be successfully used as biological control agents. 

- POSTERS -


P-38 

BACILLUS THURINGIENIS LATVIAN WIL D LW1 

INSECTICIDAL ACTIVITY AND ITS INTERACTIONS 

WITH NATURAL GUT MICROFLORA OF GYPSY 

MOTH (LYMANTRIA DISPAR) 

Z. Metla 1 , J. Halimona 1 , L. Jankevica 1 

1 Laboratory of Experimental entomology, Institute of Biology, University of Latvia, 

Salaspils, Latvia, e-mail: zmetla@inbox.lv 

Microbial populations interact with each other in different ways. Usually they are 

divided in two groups – negative interactions (competition, amensalism) and positive 

interactions (commensalism, synergism, mutalisms). Microorganisms play an 

important role in all physical processes in insects life and also have a significant effect 

on insect mortality. 

B. thuringienis is a opportunictic pathogen. There are many publications about 

interactions of B. thuringienis and natural midgut bacteria and their role in increasing 

virulence of B. thuringienis. 

We evaluated the virulence of B. thuringienis isolate LW1 on 3rd instar L. dispar 

larvae and surveyed natural midgut microflora of larvae. Depending on L. dispar 

microbial community structure, three most abudant culturable bacterial isolates were 

chosen and their mode of interactions with B. thuringienis was investigated. 

L. dispar larvae were collected from natural habitats applying standard methods. 

After collecting living insects were placed in sterile isolators for observations. 

Contents of living insects intestine were homogenized and spread on artificial 

mediums. Morphologically different bacteria were classified by carrying on necessary 

chemical reactions. Bacteria were classified applying Crystal GP identification system. 

To determine relationships between B. thuringiensis and midgut bacteria isolates, 

growth of B. thuringiensis and midgut bacteria were explored in LB broth media. 

Mortality of L. dispar larvae caused by B. thuringiensis (conc. 2x10 -7 spores/ml) 

reached 65,91% in 14 days. Composition of midgut microflora of L. dispar was 

relatively simple - it consited of six different bacteria species. Our results on 

microorganism interactions suggest that they interact positively – B. thuringienis 

didn‟t cause the decrease of population density of midgut bacteria. 

- POSTERS - 99


P-39 

ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF 

SOME NOTONECTA SPECIES (HETEROPTERA: 

NOTONECTIDAE) IN TURKEY 

100 

G. Alp Avci 1 , A, Salur 2 , E. Avci 3 , S. Coskun Cevher 4 

1 Department of Microbiology, The Hitit University, Corum, Turkey, 

e-mail: alp.gulcin@yahoo.com 

2 Department of Biology, The Hitit University, Corum, Turkey, 

e-mail: alisalur@gmail.com 

3 Department of Biology, The Hitit University, Corum, Turkey, 

e-mail: avci.emre@yahoo.com 

4 Department of Biology, The Gazi University, Ankara, Turkey, 

e-mail: sule@gazi.edu.tr 

Insects and arthropods “are a large, unexplored and unexploited source of 

potentially useful compounds for modern medicine”. To investigate the antimicrobial 

and antioxidant effects of some Notonecta species (Notonecta viridis, Notonecta 

glauca and Notonecta maculata) from aquatic Heteroptera in this study. In this 

study, N.viridis, N. maculata and N. glauca collected in Corum, Turkey. Extracts of 

N.viridis, N. maculata and N. glauca were prepared with ethanol and chloroform. The 

antimicrobial activity of extracts used in this work was tested against Escherichia coli, 

Enterococcus faecalis, Staphylococcus aureus and Candida albicans using the assay 

described by the disk diffusion method. The total antioxidant activity of extracts was 

determined using total antioxidant status (TAS) kit. Results was calculated as EQV 

mmol Trolox./L. Extracts of N.viridis, N. maculata and N. glauca have been identified 

diffrent inhibitory effect on all microorganisms. The ethanol and chloform extracts 

compared with together, ethanol extract is higher than the antimicrobial effect was 

observed. In addition, the antioxidant capacity of samples ethanol extract was 

determined that a certain level of low, albeit. The antioxidant activity of extracts 

compared with together, N. glauca extract is higher than other extracts was observed. 

These results, in terms of microorganisms in water quality by the presence of insects in 

aquatic environments suggest that there is an indication. 

References: 

1. Erel O. (2005) Clin. Biochem. 38, 1103. 

2. Pembert on, R.W. (1999) Journal of Ethnopharmacology. 65, 207. 

- POSTERS -


P-40 

DISTINCT EXPRESSION PATTERNS OF TWO 

RELATED NEUROPEPTIDES, CCHAMIDE-1 AND -2, IN 

THE SILKWORM BOMBYX MORI 

L. Roller 1 , Y. Tanaka 2 , A. Mizoguchi 3 , D. Ņitňan 1 

1Institute of Zoology, Slovak Academy of Sciences, Dúbravská 9, 84506 Bratislava, 

Slovakia., e-mail: ladislav.roller@savba.sk 

2 Division of Insect Science, National Institute of Agrobiological Sciences, 

Ibaraki 305-8634, Japan. 

3 Division of Biological Science, Graduate School of Science, Nagoya University, 

Nagoya 464-8602, Japan. 

CCHamides are recently discovered neuropetides, whose genes have been 

identified in most arthropods with a sequenced genome. In each analyzed insect two 

forms of CCHamide are encoded by separate genes. Despite characterization of Gprotein 

coupled receptors for both peptide variants in Drosophila (Hansen et al. 2011, 

Biophys. Res. Comm. 404, 184.), physiological roles for CCHamides remain elusive. 

As a prelude for further functional studies we focused on expression analysis of two 

CCHamides in the silkworm Bombyx mori (Bom). Gene transcripts encoding full 

ORFs for BomCCHa-1 and -2 were identified by PCR-based cloning and localized by 

in situ hybridization. In the silkworm, CCHa-1 and CCHa-2 are expressed in two 

distinct neuronal populations in the central nervous system (CNS) and in endocrine 

cells of the midgut. Expression patterns of both genes were slightly altered during 

metamorphosis. A group of sex-specific CCHa-1 neurons was detected in the terminal 

abdominal ganglion. Immunohistochemical analysis with antiserum against 

BomCCHa-2 revealed closer anatomy of some neurons. In pro- and mesothoracic 

ganglia a single pair of ventrolateral neurosecretory cells extends axons along the 

transverse nerves and terminates on the lateral longitudinal tracheal trunks with many 

neurohemal varicosities. CCHa-2 immunoreactivity was also detected in ~150 distinct 

interneurons distributed throughout the CNS. Widespread spatial distribution of each 

peptide form indicates its pleiotropic action in the silkworm. The distinctive 

expression patterns together with recent findings on two receptors specifically 

activated by CCHa-1 and CCHa-2 suggest that these peptides may control specialized 

physiological functions in insects. 

This work was supported by a grant from the National Institute of Health, USA (GM 67310), and 

Slovak grant agencies, Agentúra na podporu výskumu a vývoja (APVV-51-039105) and Vedecká 

grantová agentúra (VEGA 2/0132/09). 

- POSTERS - 101


P-41 

EFFECTS OF QUININE, QUINIDINE 

AND DENATONIUM ON FEEDING BY CODLING MOTH 

NEONATES 

102 

Maciej A. Pszczolkowski 

William Darr School of Agriculture, Missouri State University, 

Mountain Grove, MO 6571, USA, e-mail: MPszczolkowski@missouristate.edu 

Effects of bitter tasting compounds on feeding behavior of neonate larvae were 

studied in codling moth, Cydia pomonella. Preliminary results show that denatonium 

delays feeding commencement and reduces amounts of food consumed. Quinine and 

quinidine at concentrations up to 0.5 mg/ml did not have significant effects on feeding 

behavior of the larvae. Exposure of the larvae to food treated with denatonium and 

phospholipase C inhibitor suggests that phospholipase C is involved in transduction of 

the signal from denatonium. 

- POSTERS -


P-42 

CRANGON CRANGON ECDYSONE RECEPTOR AND 

RETINOID-X-RECEPTOR COMPLEX: DIMERIZATION 

INFLUENCED BY LIGAND BINDING? 

Ruben De Wilde a,b , Luc Swevers c , Yves Verhaegen a,b , Pierre Rougé d , Kris Cooreman b , 

Guy Smagghe a 

a Laboratory of Agrozoology, Faculty of Bioscience Engineering, Ghent University, 

Coupure Links 653, B-9000 Ghent, Belgium 

b Institute for Agricultural and Fisheries Research, Animal Sciences Unit, Fisheries 

Department, Ankerstraat 1, B-8400 Ostend, Belgium 

c Insect Molecular Genetics and Biotechnology, Institute of Biology, National Centre 

for Scientific Research “Demokritos”, Aghia Paraskevi Attikis, Athens, Greece 

d Surfaces Cellulaires et Signalisation chez les Végétaux, UMR Université Paul 

Sabatier CNRS 5546, Castanet Tolosan, France 

Many crustaceans inhabiting the shallow coastal shelf zone, are under heavy 

environmental pressure of human activities. A good example is the common brown 

shrimp (Crangon crangon L.) in the North Sea and Wadden Sea. In this project we 

report on the interaction between the ecdysone receptor (EcR) and the retinoid-Xreceptor 

(RXR) of C. crangon with use of a (yeast-derived) Gal4 reporter system. 

These two important nuclear receptors play a crucial role in growth, development and 

reproduction of crustaceans. EcR binds the ecdysteroid hormone and forms a 

heterodimer with RXR, starting the hormone signalling cascade. It is evident that any 

impairment of EcR/RXR function results in abnormalities and eventually death of the 

organism. Based on previous cloning of these two hormone receptors (Verhaegen et al. 

2010, 2011), we prepared truncated (consisting of the hinge and ligand binding 

domains) and full length chimeric constructs of CrcEcR and CrcRXR, fused to the 

heterologous Gal4-activating domain (AD) and Gal4-DNA binding domain (DBD). 

These constructs were co-transfected into insect Hi5 cells together with a luciferase 

reporter plasmid under control of a (4x) Gal4-upstream activation sequence (UAS). 

Here we report on the homo- and heterodimerization of the CrcEcR and CrcRXR in 

insect cells, and discuss on the importance of natural ecdysteroid hormone ligands 

(Ponasterone A and 20-hydroxyecdysone) and the effects of important contaminants as 

tributyltin (TBT). 

Keywords: Crangon crangon, EcR, RXR, Gal4, dimerization, ligand 

Verhaegen, Y., Parmentier, K., Swevers, L., Rougé, P., Soin, T., De Coen, W., Cooreman, K. & 

Smagghe, G. (2010). The brown shrimp (Crangon crangon L.) ecdysone receptor complex: cloning and 

functional expression in an EcR-deficient Drosophila cell line. General and Comparative 

Endocrinology, 168(3): 415-423. 

Verhaegen, Y., Parmentier, K., Swevers, L., Renders, E., Rougé, P., De Coen, W., Cooreman, K. & 

Smagghe, G. (2011). The heterodimeric ecdysteroid receptor complex in the brown shrimp Crangon 

crangon: EcR and RXR isoform characteristics and sensitivity towards the marine pollutant tributyltin. 

General and Comparative Endocrinology, in press. 

- POSTERS - 103


P-43 

INFLUENCE OF DIGOXIN – ANTIARRHYTHMIC DRUG 

– ON THE HEART CONTRACTILE ACTIVITY OF 

ZOPHOBAS ATRATUS BEETLE 

M. Szymczak, M. Kowalski, P. Marciniak, J. Pacholska-Bogalska, E. Czarniewska, 

G. Rosiński 

104 

Department of Animal Physiology and Development, Adam Mickiewicz University, 

Umultowska 89 Str., 61-614 Poznan, Poland, e-mail: monikasz@amu.edu.pl 

Researches on insect heart can contribute to increasing our knowledge about 

mechanisms of drug influence on functioning of this organ. Data from evolutionary 

and comparative biology shown that organogenesis of animal heart and basic 

physiology of myocytes differ insignificantly even between distant systematic groups. 

We studied action of digoxin, drug from glycoside group, on the semi-isolated 

heart of Zophobas atratus beetle. The studies showed that digoxin applied at 

concentration range of 10 -11 -10 -5 M exerts a dose-dependent positive chronotropic 

action and reduces arrhythmia in this beetle. However, after washing out this drug 

from incubation system and returning to perfusion with normal physiological saline 

arrhythmia was observed in the contractile activity of myocardium. Terfenadineinduced 

arrhythmia in the heart contractions was abolished after application of 

digoxin. These studies shown new, unexpected action of digoxin. We confirm that 

glycoside interacts with receptor(s) on the myocardium according to Clark‟s 

hypothesis. 

Our studies showed that when we analyze even good known drug on isolated 

organ, we can get new information about it. We can define drug action and we can 

avoid its influence on another systems in organism. 

Grant support: N N303 810 640 (National Science Centre) 

- POSTERS -


P-44 

SUSCEPTIBILITY TO ORGANOPHOSPHATE 

INSECTICIDE IN TWO PEST SPECIES: WESTERN 

CORN ROOTWORM (DIABROTICA VIRGIFERA 

VIRGIFERA Le Conte) AND COLORADO POTATO 

BEETLE (LEPTINOTARSA DECEMLINEATA Say) 

M. Tarnawska 1 , M. Nakonieczny 1 , I. Kalita 1 , A. Jeleń 1 , P. Migula 1 , P. Bereś 2 , S. 

Drzewiecki 3 

1 Department of Animal Physiology and Ecotoxicology, University of Silesia, 

Katowice, Poland, e-mail: monika.tarnawska@us.edu.pl 




Organophosphate compounds (OP) inhibit esterase activity by irreversible 

binding to the catalytic site of enzymes. Acetylcholinesterase (AChE, EC 3.1.1.7) is an 

enzyme playing a key role in the modulation of neuromuscular interactions. 

Carboxylesterases (CbEs, EC 3.1.1.1) are responsible for the initial phase of enzymatic 

detoxification. They contribute to an efficient protective mechanism against acute 

toxicity of organophosphate pesticides in homeotherms and are involved in the 

biochemical mechanism of pesticide resistance in many pest insect species. 

Susceptibility to dimethoate (OP) was assessed by AChE and CbE (biomarkers of 

OP toxicity) activity measurements in two insect pest species: Western corn rootworm 

(WCR) and Colorado potato beetle (CPB). The first one become recently a serious pest 

of corn crops in Poland and the data about its exposition to dimethoate (OP) are still 

lacking. Pesticide was applied topically in a dose 0.15 µg/100 mg fresh weight per 

adult (LD50) to both species under controlled laboratory conditions. Activity of 

enzymes was evaluated in homogenates isolated from the whole body 1 hour and 24 

hours after pesticide administration and compared with the appropriate data of 

untreated insects. 

Differences in activity patterns of both enzymes were species dependent. 

Activities of tested esterases were lover in CPB than in WRC – up to 20 times for 

AChE and 3-4 times for CbE, depending on a pesticide treatment. Only in case of 

WRC a slight, insignificant, rise of AChE and CbEs activity 1 hour after and a 

significant inhibition 24 hours after the pesticide treatment were demonstrated. Higher 

activity of both enzymes in WCR indicates its higher susceptibility to the tested 

insecticide in laboratory conditions. Bearing in mind feeding behavior of this species 

we suppose that in this stage dimethoate might be applicable for chemical control of 

D. virgifera. 

- POSTERS - 105


P-45 

PROTEOMICS ANALYSIS IN GUT OF SUNN PEST, 

EURYGASTER INTEGRICEPS (PUT): ADULT INSECTS 

AND FIFTH-INSTAR NYMPHS 

106 

Mohammad Saadati Bezdi 1* , Reza Farshbaf Pourabad 1 , Mahmoud Toorchi 1 , 

Nosratollah Zarghami 2 , Setsuko komatsu 3 , Mirhasan Hashemi 4 

1 Faculty of Agriculture, University of Tabriz, Tabriz, Iran. 

Mohammad.saadatibezdi@gmail.com 

2 Faculty of Medicine, Tabriz University of Medical sciences, Tabriz, Iran, 

zarghami@yahoo.com 

3 National Institute of Crop Science, Tsukuba 305-8518, Japan, skomatsu@affrc.go.jp 

4 Jahade-keshavarzy organization, Tabriz, Iran. 

Gut of insect is main responsible of feeding which has a critical role in insect 

life and any disturbance in this process affects the other physiological processes such 

as metamorphosis and fecundity. Gut of one-day-old adult insects and fifth-instar 

nymphs after feeding from wheat, were dissected from body for proteomics analysis. 

Crude proteins extracted from guts were separated by 2-DE and visualized by CBB. 

After image analysis, 23 protein spots which differently changed between adult insects 

and fifth-instar nymphs were cut and subjected to in gel trypsin digestion and then 

peptides were injected to nano LC-MS/MS. Peptide masses were searched in NCBI 

database using mascot search engine. Tropomyosin, actin and arginin kinase which 

related to mechanical digestion; glutamate dehydrogenase, glyceraldehide-3-phosphate 

dehydrogenase, enolase, acyl-CoA dehydrogenase and ATP syntheas as metabolismic 

proteins and defense proteins such as catalase and translationally controlled tumor 

protein were accumulated in adult insects. Three plant proteins such as serpin, 

dehydroascorbate reductase and beta-amylase were accumulated in gut of adult insects. 

Heat shock protein 70 was increased in gut of fifth-instars nymphs. These results 

proposed that efficiency of digestive processes and defense response in gut of adult 

insects was more advanced in comparison to fifth-instar nymphs. This study was 

performed for first times in world and we hope to complete our results in near future. 

- POSTERS -


P-46 

CENTRAL IMMUNOMODULATORY EFFECT OF 

SYNTHETIC ALLOFERON 1 (Al 1) IN RATS 

Rykaczewska-Czerwińska Monika 1 , Oleś Piotr 1 , Oleś Michał 1 , Brewczyński Adam 1 , 

Kuczer Mariola 2 , Konopińska Danuta 2 , Plech Andrzej 1,3,4 

1 Department of Pharmacology, Medical Faculty in Zabrze of the Medical University of 

Silesia in Katowice, 28 Jordana Str., 41-808 Zabrze, Poland, 

e-mail: monczer@gazeta.pl 

2 Faculty of Chemistry, Wrocław University, 14 F Joliot-Curie Str., 

50-383 Wrocław, Poland 

3 Cardinal August Hlond Upper Silesian College of Pedagogical Education, Faculty of 

Public Health, 19 Powstańców Str., 41-400 Mysłowice, Poland 

4 Silesian College of Economics and Administration, Faculty of Physiotherapy, 12 

A.Frycza – Modrzewskiego St., PL 41-907 Bytom, Poland 

Insects have a potent defensive mechanisms against microbial infections, by 

producing various agents, including antibacterial peptides. Two bioactive peptides 

were isolated from hemolymph of experimentally infected larvae of the blow fly 

Calliphora vicina (Chernysh S. et al. 2002, Proc. Natl. Acad. Sci USA, 99, 12628- 

12632). These peptides were named as alloferons, due to their functional similarity 

with interferon (IFN) (Chernysh S. et al. 2002, Proc. Natl. Acad. Sci USA, 99, 12628- 

12632). The largest, 13-residue, extracted peptide was named alloferon 1 

(HGVSGHCQHGVHG). It was reported that synthetic alloferon 1 stimulates activity 

of NK lymphocytes from mouse spleen and from human peripheral blood. Moreover it 

stimulated IFN in mice, as well as displayed antiviral and antitumoral effect 

(Chernysh S. et al. 2002, Proc. 

The study presented in this report were undertaken in order to evaluate 

immunomodulatory effect of alloferon 1 administered directly into the lateral brain 

ventricle of the rat. A week before experiments rats were anaesthetized with ketamine 

and xylazine and polyethelene cannulas were stereotaxically implanted into the lateral 

brain ventricle. On the day of experiment synthetic Al 1 was injected into the lateral 

brain ventricle (icv) through implanted cannulas at doses of 5, 25, 50 and 100 nmols. 

Every dose of Al 1 was dissolved in a constant volume of 5 μl of 0.9% NaCl. Control 

animals were treated with the same volume of alone 0.9% NaCl. After 2 and 24 h after 

icv administration of Al 1 or saline, rats were again anaesthetized and a sample of the 

blood was taken by heart puncture. Plasma levels of IFN-γ, TNF-α and Il-2 were 

determined by ELISA method. Al 1 induced a slight decrease of IFN-γ and a 

significant decrease of TNF-α levels. 

Obtained results indicate that intracerebrally (i.e. icv) applied Al 1 may 

influence on cytokine levels in peripheral blood in rats. At present the mechanism of 

this central effect of Al 1 is unknown. We regard that central nervous system at least in 

part may mediate immunomodulatory effect Al 1, expressed in changed cytokine 

levels. 

Acknownledgements: This work was supported by a grant from the Medical University of Silesia 

KNW-2-031/10 and by a grant from Ministry of Science and Higher Education N N 204085638. 

- POSTERS - 107


P-47 

ARTHROPODS INFESTATION DETECTION IN ACACIA 

SEYAL VARIETY SEYAL (BRENAN) NATURAL STANDS 

IN KORDOFAN REGION, SUDAN 

108 

Maymoona Ahmed Eisa 1, 2 , Mohammed Hamed Mohammed 2,3 , Ingo Brunk 1 , 

Heinz Röhle 3 

1 Technische Universität -Dresden, Institute of Forest Botany and Forest Zoology, 

Tharandt, Germany, Fax: +49 (0) 35203-38-31317 

email: maymoonaeisa@yahoo.com 

2 University of Kordofan, Faculty of Natural resources & Environmental Studies, P.O 

Box 160 -Elobeid 51111, Sudan 

3 TU-Dresden, Institute of Forest Growth and Computer Sciences, Tharandt, Germany 

Corresponding author: M. A. Eisa 1 

Acacia seyal variety seyal is an important economic tropical dry savannah tree 

producing fuel-wood and gum which has a significant contribution to total production 

of gum Arabic in Sudan. The tree species has undergone insects‟ damage that may has 

influence in gum yield. This paper presents the damage exerts by some arthropods 

species on Acacia seyal variety seyal natural stands in Kordofan region, Sudan. In 

season 2007/2008, a field survey was conducted in Umfakarin natural forest reserve, 

south Kordofan. Trees of Acacia seyal were selected based on their diameters at breast 

height (DBH, 5-40 cm) mainly for the purpose of gum production experiment. During 

the field survey, the existence and non-existence of tree damage by Isoptera and 

Coleoptera: Bostrichidae were reported. Data were analysed using a binary logistic 

regression. The results indicate that DBH was found to be the only relevant predictor 

influencing tree infestation. The probability of tree infestation increases when diameter 

increases. Further investigation is needed to determine the infestation predictors in 

order to draw effective control measures of Acacia seyal insect pests. 

Keywords: Acacia seyal, Isoptera and Coleoptera, logistic regression, Umfakarin forest. 

- POSTERS -


P-48 

STUDIES ON THE EFFECT OF MICROWAVES 

RADIATION AND COLD STORAGE ON ORYZAEPHILUS 

SURINAMENSIS ( COL. : SILVANIDAE) 

AND SITOPHILUS ORYZAE ( COL. :CURCULIONIDAE) 

ADULTS 

Nouraddin Shayesteh 1 , Somayyeh Ghasemzadeh 2 , Ali A. Pourmirza 2 , Mohammad H. 

Safaralizadeh 2 , Mustafa Maroufpoor 2 

1. Department of Plant Medicine, Mahabad Branch Islamic Azad University, 

Mahabad-Iran 

2. Department of Plant Medicine, Agriculture faculty, University of Urmia, IRAN 

Corresponding Author E-mail: profshayesteh@yahoo.com, Mob: +989141414206, 

Fax:+984422336000 

The combined impact of microwave radiation and cold storage on Oryzaephilus 

surinsmensis (Herbst) and Sitophilus oryzae (L.) adults either continuously or 

intermittently was evaluated. The insects were exposed to 2450 MHz at power level of 

100 W for exposure time of 10 min continuously and intermittently. In all experiments, 

the highest rate of mortality was achieved for exposure time of 10 min intermittently 

exposure and 72 h cold storage duration. Intermittent exposures were generally more 

effective in killing insects of both species compared with those of continuous 

irradiation. Combinations of microwave radiation and cold storage were found highly 

compatible and synergistic. The synergistic interaction indicates that microwave 

radiation can be used with cold storage for management of Oryzaephilus surinsmensis 

and Sitophilus oryzae adult developmental stage. This treatment could provide an 

effective and friendly environmental treatment technique in IPM program. 

Keyword: microwave irradiation, temperature manipulation, Oryzaephilus surinamensis and Sitophilus 

oryzae 

- POSTERS - 109


P-49 

EVALUATION OF MICROWAVES RADIATION AND 

COLD STORAGE ON MOLECULAR PHYSIOLOGY OF 

STORED-PRODUCT INSECTS AND WHEAT SEED 

Nouraddin Shayesteh 1 , Somayyeh Ghasemzadeh 2 , Ali A. Pourmirza 2 , Mohammad H. 

Safaralizadeh 2 , Mustafa Maroufpoor 2 

1. Department of Plant Medicine, Mahabad Branch Islamic Azad University, 

Mahabad-Iran 

2. Department of Plant Medicine, Agriculture faculty, University of Urmia, IRAN 

Corresponding Author E-mail: profshayesteh@yahoo.com, Mob: +989141414206, 

Fax:+984422336000 

Tribolium castaneum and Oryzaephilus surinamensis which are two main 

stored-product insects were exposed to 2450 MHZ frequency at level power 100 W for 

10 min continuously and intermittently. The results showed that the order of 

susceptibility of insects to microwave radiation was as follows: O. surinamensis > T. 

castaneum. Intermittent exposures were generally more effective in killing insects of 

both species compared with those of continuous irradiation. Combinations of 

microwave radiation and cold storage were found highly compatible and synergistic. 

In order to have some insights on the impact of microwave power on wheat 

germination experiments were carried out according to standard protocols. We used 

100 and 200 W of microwave power for 2, 4, 6 and 10 min. We found a drastic 

reduction in germination rate under 200 W microwave powers and 10 min. exposure 

time. 

Keywords: Microwave radiation, Cold storage, Stored-product insects 

110 

- POSTERS -


P-50 

RELATIONSHIP OF THE QUANTITATIVE AND 

QUALITATIVE VOLATILE OIL CONTENTS OF CITRUS 

LEAVES WITH INFESTATION OF CITRUS VARIETIES 

WITH CITRUS LEAF MINER PHYLLOCNISTIS 

CITRELLA 

Mogahed M.I.(1), N. Nazif(2), Kh.A. Abdel Shafeek(2) and M.M. El-Missiry(2) 

(1) Pests and Plant Protection Dept., NRC, Dokki, Cairo, Egypt 

E-mail: mogahedprof2007@hotmail.com 

(2) Chemistry of Medicinal Plants Dept.,NRC 

Screening studies carried out on 7 citrus varieties for their susceptibility to 

infestation with Phyllocnistis citrella Stainton at Qualiyobiya and North Sinai 

Governorates, Egypt, highest infested citrus varieties were Lemon, Grapefruits and 

Saify followed by navel and mandarin. The least infested varieties were sweet and 

baladi orange. Infestation with P. citrella were seasonally fluctuated. Leaves 

infestation (%) were larger at Qualiyobiya than North Sinai. 

Odorous constituents such limonene (Terpene hydrocarbon) in lime. Acids and 

esters of organic acids, in grapefruit making its leaves more attractive to insect than 

other varieties. It was concluded that total content of volatile oils in citrus leaves were 

significantly affected by diversity of varieties. Differentiation of citrus infestation with 

CLM depends on total content of V. oils in citrus leaves and levels of some odorous 

constituents. There is a relationship between chemical composition of V.O. and 

infestation with CLM. Limonene, linalool (T. alcohol) and esters of organic acid in V. 

oils content increased insect infestation of lemon, grapefruits, saify and mandarin 

varieties. An increase of aldehydes and ketones in V. oils content decreased infestation 

of baladi and navel trees caused with P. citrella. 

Keywords: Citrus varieties –Phyllocnistis citrella – volatile oils. 

- POSTERS - 111


AUTHOR INDEX


-AUTHOR INDEX- 113

114 


Abdel Shafeek Kh.A. P-50 

Abdel-Latief M. PL-2 

Abdel-Wali M. O-25 

Abdin Z. O-27 

Adams M.E. O-33 

Affifi A. O-37 

Ahadiyat A. O-41 

Ahmed Eisa M. O-35, P-47 

Ahsan Khan M. O-5 

Ajam Hassani M. O-4 

Al-Antary T. O-25 

Alipour E. P-29 

Alp Avci G. P-39 

Ameneh Asadi P-34, P-35 

Arensburger P. PL-3 

Arshad M. O-5, O-6, O-27 

Asaoka K. O-26 

Ashfaq M. O-5 

Askari G. O-19 

Ateyyat M. O-25 

Atkinson P.W. PL-3 

Audsley N. O-46 

Augustyniak M. P-12 

Avci E. P-39 

Babmorad M. O-28 

Bano M. O-44 

Baruah A.A.L.H. O-24 

Bayhan E. P-26, P-27 

Belhadj S. P-18 

Bereś P. P-13, P-14, P-44 

Bertossa M. P-30 

Bhattacharyya B. O-24 

Bhuyan U. O-24 

Bissaad Z. P-16, P-17 

Blenau W. O-39 

Boerjan B. O-10 

Borovsky D. O-3, O-4 

Bounaceur F. P-16, P-17 

Bousaad F. P-18 

Bretagnolle V. O-12 

Breuer M. P-31 

Breugelmans B. O-38, O-40 

Brewczyński A. P-46 

Brown J.J. O-14


Brunk I. O-35 

Brunk I. P-47 

Burrows M. O-39 

Casartelli M. P-2, P-3 

Castelli I. P-2, P-3 

Chakravorty J. O-33 

Chebouti-Meziou N. P-18 

Chrzanowski G. P-19, P-23, P-28 

Cieniecka-Rosłonkiewicz A. P-20 

Coast G. PL-1 

Coissac E. O-12 

Cooreman K. P-42 

Coskun CevherS. P-39 

Cowell B. O-14 

Craig N. L. PL-3 

Czarniewska E. P-21, P-22, P-43 

Czerniewicz P. P-11, P-19, P-23 

Dancewicz K. P-24 

Dastour L. P-25 

Daubnerová I. O-32 

De Wilde R. P-42 

Denlinger D. PL-1 

Diamante B. P-2, P-3 

Djadid N D. O-8 

Doumandji S. E. P-18 

Doumandji-Mitiche B.P-16, P-17 

Down R. O-46 

Drzewiecki S. P-13, P-14, P-44 

Ebrahimi L. O-19 

Ebrahimzade M. P-25 

El-Gengaihi S. E. O-37 

El-Missiry M.M. P-50 

Esmaiilizadeh M. O-31 

Fahim udin Shah H. O-27 

Farazmand H. O-43 

Farshbaf Pourabad R. O-19, O-42, P-25, 

P-29, P-45 

Faßold K. O-7 

Fekri M. O-31 

Gabryś B. P-24 

Gadwala M. O-23 

Gauffre B. O-12 

Ghadamyari M. O-4, P-8 , P-34, P-35 

Ghasemzadeh S. P-49, P-48 

Giordana B. P-2 

Gliszczyńska A. P-24 

Gogi D. O-5 

Goławska S. P-11, P-19, P-28 

Gorzka D. O-30 

Gosselin Grenet A.S. P-3 

Greiner A. P-31 

Grudniewska A. P-24 

Guy Smagghe G. O-11 

Guzik J. P-13, P-14 

Hagiya H. O-26 

Hajizadeh J. P-8 

Halimona J. P-37, P-38 

Hamed Mohammed M.P-47 

Hamshou M. O-11, P-45 

Hata T. O-26 

Hecker E. P-32 

Hein D.F. P-31 

Hellmann C. P-31 

Hesamzadeh M. O-28 

Hice R.H. PL-3 

Hiiesaar K. P-33 

Hoffmann K. PL-2 

Hołdaj M. O-30 

Horowitz A. R. O-18 

Hosseinzadeh J. O-43 

Hummel H.E. O-36, P-30, P-31, 

P-32 

Huybrechts R. O-1, O-2, O-10 

Inoue T.A. O-26 

Inui H. O-2 

Ishaaya I. O-18 

Ismail I. A. O-37 

Ito T. O-26 

Jafarlu M. O-42 

Jalal Arif M. O-5 

Jalal Jalali P-34, P-35 

Jalali Sendi J. O-4, P-8 

Janda B. P-28 

Jankevica L. P-37, P-38 

Jedlička P. P-7 

Jeleń A. P-44 

Jõgar K. P-33 

Kaczmarek K. PL-1 

Kaissling K.E. P-32 

Kalita I. P-44 

Kaliwal B.B. O-23 

Kamenova S. O-12 

-AUTHOR INDEX- 115

116 


Kapusta I. P-28 

Kari N. O-23 

Karimpour Y. O-43 

Kazimierczak J. P-20 

Kędziorski A. P-12 

Kevin Durden K. O-14 

Kindl J. P-5 

Kivimägi I. P-33 

Kleeberg H. P-31 

Kluczyk A. O-16 

Kodrík D. O-34, P-5 

Komatsu S. P-45 

Konopińska D. O-29, P-9, P-10 

P-46 

Kowalski M. P-43 

Kownacki A. O-20 

Kozak A. P-19 

Kratt A. P-31 

Kripach V. O-9 

Kuczer M. P-9, P-10, P-21, 

P-22, P-46 

Kuusik A. P-33 

Łaszczyca P. P-1 

Lebedev G. O-18 

Leithold G. P-31 

Leszczyński B. P-11, P-19, P-23 

Lindemans M. O-1 

Lindner I. P-31 

Litvinovskaya R. O-9 

Lorenz M.W. O-7 

Łuczak M. P-10 

Łukasik I. P-28 

Mahmoud P-45 

Majewska A. P-10 

Mänd M. P-1, P-33 

Manzoor A. O-5 

Marciniak P. P-9, P-15, P-43 

Mari J.M. O-15 

Maroufpoor M. P-49, P-48 

Matthews J. O-46 

Mehdi M. M. O-41 

Merivee E. P-6 

Metla Z. P-37, P-38 

Metspalu L. P-33 

Meyering-Vos M. PL-2 

Meyer-Rochow V.B. O-33 

Michailova P. O-20 

-AUTHOR INDEX- 

Michalczyk A. P-20 

Midak-Siewirska A. P-10 

Migula P. P-12, P-13, P-14, 

P-44 

Mizoguchi A. P-40 

Mogahed M. I. O-21, P-50 

Moger N. O-23 

Mohammadi D. O-42, P-25, P-29 

Morytz B. P-20 

Mrówczyńska L. P-21, P-22 

Mubarak E. O-5 

Muljar R. P-1 

Must A. P-6 

Nachman R. PL-1 

Nagaba Y. O-1, O-2 

Nakonieczny M. P-13, P-14, P-44 

Nazif N. P-50 

Niihara K. O-26 

Nocoń Ł. P-12 

Nur Azura A. O-45 

Ogliastro M. P-3 

Oleś M. P-46 

Oleś P. P-46 

Ölmez Bayhan S. P-26, P-27 

Olszak R. W. O-30 

Ott S.R. O-39 

Pacholska-Bogalska J.P-9, P-15, P-43 

Pennacchio F. P-2 

Plantegenest M. O-12 

Plech A. O-29, P-46 

Ploomi A. P-33 

Pourmirza A.A. P-49, P-48 

Pszczolkowski M.A. O-14, P-41 

Pujari D. O-24 

Reza Hassan Sajedi P-34, P-35 

Rifat Ulusoy M. P-27 

Rogalska L. P-9 

Röhle H. P-47 

Roller L. O-32, P-40 

Rosiński G. P-9, P-15, P-21, 

P-22, P-43 

Rougé P. P-42 

Rouhani M. O-31, P-36 

Rykaczewska-Czerwińska M.O-29, P-46 

Saadati Bezdi M. P-45 

Sabbour M. O-22


Sadeghi S.E. O-28 

Saeidi K. O-45 

Safaralizadeh M.H. P-49, P-48 

Salur A. P-39 

Samantha Sellars S. O-14 

Samih M. A. O-31, P-36 

Sarafrazi A. O-41 

Satar Z. O-28 

Saud R.K. O-24 

Saxena M.M. O-13 

Schneedorferová I. P-5 

Semenov A. P-4 

Seskena R. P-37 

Shaina H. O-5 

Shayesteh N. O-43, P-49, P-48 

Shyam Kumar V. O-23 

Ńimek P. P-7 

Simonet G. O-40 

Skorek A. P-14 

Slama K. O-9 

Smagghe G. PL-1, P-42 

Socha R. O-34 

Sohail A. O-5 

Solhjouyefard S. O-41 

Spit J. O-40 

Sprawka I. P-11, P-23, P-28 

Srivastava M. O-13 

Steinbauerová V. P-7 

Suhail A. O-5, O-6, O-27 

Ńvilponis E. P-33 

Swevers L. P-42 

Sytykiewicz H. P-11, P-19, P-23 

Szarek-Gwiazda E. O-20 

Szymczak M. P-9, P-15, P-43 

Taberlet P. O-12 

Tahir M. O-44 

Tajeddin L. O-8 

Takeda M. O-2 

Tanaka S. O-17 

Tanaka Y. P-40 

Tarnawska M. P-13, P-14, P-44 

Tobback J. O-10 

Tomčala A. O-34, P-5 

Tooming E. P-6 

Tufail M. O-2 

Valizade M. O-42, P-29 

Valterová I. P-5 

Van Damme E.J.M. O-11 

Vanden Broeck J. O-38, O-39, O-40 

Vandersmissen H.P. O-10 

Vannini L. PL-2 

Verhaegen Y. P-42 

Verlinden H. O-1, O-38, O-39 

Veromann E. P-33 

Vinokurov K. O-34 

Vleugels R. O-39 

Vuerinckx K. O-1 

Warchałowska-Śliwa E. O-20 

Wawrzeńczyk Cz. P-24 

Wende F. PL-2 

Wendorff J.H. P-31 

Woodring J. PL-2 

Woźniak K. P-15 

Wright J.A. PL-3 

Wynant N. O-38 

Yazdani E. P-8 

Yokohari F. O-26 

Zabrocki J. PL-1 

Zahorska R. P-10 

Zain ul Abdin O-5 

Zakeri S. O-8 

Zarghami N. P-45 

Zarins I. P-37 

Zhylitskaya H. O-9 

Ņitňan D. O-32, P-40 

-AUTHOR INDEX- 117


118

concentration of polysaccharides in spring - vii-th international ...

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?