Mycotoxin E book 3 A5 size 04

MYCOTOXIN
&
PESTICIDE
Digest For Poultry
Kemin Industries South Asia Private Limited
#C-3, First Street, Ambattur Industrial Estate, Chennai - 600 058, INDIA
044 4220 2800 | mail.india@kemin.com | www.kemin.com
TM
© Kemin Industries, Inc. and its group of companies 2017 All rights reserved. ® Trademarks of Kemin Industries, Inc., U.S.A.
zestcreative.in

MYCOTOXIN
&
PESTICIDE
Digest For Poultry
Kemin Industries South Asia Private Limited
#C-3, First Street, Ambattur Industrial Estate, Chennai - 600 058, INDIA
044 4220 2800 | mail.india@kemin.com | www.kemin.com
TM
© Kemin Industries, Inc. and its group of companies 2017 All rights reserved. ® Trademarks of Kemin Industries, Inc., U.S.A.
zestcreative.in

INDEX
Mycotoxins
What Are mycotoxins? 6-7
Where do mycotoxin come from? 8-9
What are different kinds of mycotoxins? 10 -11
Difference between molds and mycotoxins? 12-13
Critical points in c production- New Maize 14-15
Critical points in mycotoxin production- Field and Storage 16-17
What are the factors affecting mycotoxin production? 18-19
Mycotoxin Prevalence and shi in contamination 20-21
What is the safe limit? 22-23
How do they affect poultry health and production? 24-25
Pesticides
What are Pesticides? 28
What is pesticide toxicity? 29
How pesticides enter bird’s body? 30-31
What is MRL? 32-33
What is the effect of chemical toxicity in poultry? 34-35
Analysis of Mycotoxins
What are different methods to analyze ycotoxin? 36-37
Prevention
How can we prevent mycotoxin contamination in feed? 40-41
Why is Sampling a critical point? 42-43
What is the proper sampling method? 44-45
o
Kemin’s 360 toxin
Management program
2 3

MYCOTOXINS
4 5

What are Mycotoxins?
• Mycotoxins are the secondary metabolites of certain molds
which occur on agricultural products. Various mycotoxins
have been studied for their toxic effects on both humans
and animals.
• Even the minimalist quantity of mycotoxins is harmful in
terms of productivity, immunosuppression etc.
Aflatoxin B1 | Ochratoxin A |
Zearalenone (ZON) | T-2 Toxin
Fumonisin B1 | Deoxynivalenol (DON)
Cyclopiazonic Acid (CPA)
Mycophenolic Acid (MPA)
J. W. Bennett1,* & M. Klich2
6 7

Where do Mycotoxin
come from?
• Mycotoxins are produced by toxigenic fungus or molds
• The reason for their production is still unknown but it is
believed that they help them to survive in the niche
environment
• One Fungus can produce more than one type of mycotoxins
and vice versa
Myth #1
USING MOLD INHIBITOR IN FEED WILL
SOLVE MYCOTOXIN PROBLEM
Myth Buster
False. Mycotoxins produced before application of mold
inhibitor will still be present and are not destroyed.
J. W. Bennett1,* & M. Klich2
8 9

What are different
kinds of Mycotoxins?
Classification Mold Mycotoxins
Field Fusarium sp. Fumonisin B1, T-2
Toxin, Deoxynivalenol
(DON), Zearalenone
(ZON)
Storage Penicillium sp. Ochratoxin A,
Cyclopiazonic acid
(CPA), Citrininn
Aspergillus Sp.
Cyclopiazonic
Acid(CPA), Aflatoxin
B1, B2, G1, G2,
Ochratoxin A
http://www.fao.org
10 11

MYCOTOXINS
Difference between
Molds and Mycotoxins?
• Molds are filamentous fungi which produce secondary
metabolites called mycotoxins.
• Molds grow on cereals and grains commonly used in feedmaize,
wheat etc.
• In the challenge period, fungus produces mycotoxins to
survive in the niche environment .
• Different fungus may produce different or same
mycotoxins depending on the environment temperature,
moisture etc.
• Molds can be destroyed by on feed application of mold
inhibitors but not mycotoxins.
• Mycotoxin once entered in bird’s digestive system cannot
be destroyed and are required to be excreted.
Molds
Nutrient reduction in feed
Reduce palatability of feed
Change in physical
appearance of feed
Invite insects on feed
which may carry
pathogenic organisms
E.g. Aspergillus flavus,
Penicillium etc.
Mycotoxins
Impacts performance of
the bird negatively
In high concentration
cause mortality
Affects liver, kidney, immune
system etc. of the bird
Increases organ weight
E.g. Aflatoxins, Fumonisins,
Richard S. Adams,
Maciorowski et al., 2007
12 13

MYCOTOXINS
Critical points in
Mycotoxin production
In India, there are two
major seasons of
maize harvest
1. March to May- Bihar
2. October to
January- Karnataka
Generally maize
is harvested
at 20-25% moisture
It is then dried
for 1 week to bring down
moisture to 10-12%
In monsoons,
harvest is not able to
dry properly
Because of this inadequate
loss of moisture, it remains
at 15-17%. This invites
high fungus growth
and subsequent
mycotoxin production
https://www.indiaagronet.com/indiaagronet/crop%20info/maize.htm
14 15

MYCOTOXINS
Critical points in
Mycotoxin production
Field Injuries:-
Insect injuries,
mechanical injuries
Improper drying
before transport
(Moisture >12%)
Storage at both
farmer and feed
producer
16 17

MYCOTOXINS
What are the factors
affecting Mycotoxin
production?
Mycotoxin
producing fungus
Mycotoxin produced
Climate condition
Optimum
Temp Reqt.
Optimum
Water Activity
Aspergillus flavus, A. parasiticus,
A. pseudotamarii
Aflatoxin B1, B2, G1, G2 (Afla)
Warm, Humid, Tropical,
Delayed harvest, >50mm rainfall
0
33 C
0.99
A. ochraceus, Penicillium verrucosum
Ochratoxin A (OTA)
High moisture on harvest, Damp storage
0
25-30 C
0.98
A. flavus A. versicolor,
Penicillium cyclopium,P. camemberti
Cyclopiazonic Acid(CPA)
High moisture subtropical conditions
0
20 C
0.98
Fusarium verticillioides,
F. proliferatum
Fumonisin B1 (FUMB1)
Temperate conditions
0
15-30 C
0.90-0.99
F. graminearum F. avenaceum
Trichothecenes: T-2 toxin,
High Moisture and Subtropical areas
0
26-30 C
0.995
F. graminearum
Trichothecenes: Deoxynivalenol (DON)
Sensitive to slightest
change in temperatures
0
26-30 C
0.995
F. graminearum, F. sporotrichioides
Zearalenone (ZON)
A humid tropical climate is ideal
0
25 C
0.96
Penicillium citrinum, P. verrucosum
Citrinin
Generally formed post harvest
in stored grains
20-30 0
C
0.75-0.85
J.M. Milani PAUL E. NELSON et al Wageha Awad et al A.GENTLES et al Jewers K.
18 19

MYCOTOXINS
Prevalence
in Asia
900
800
700
600
500
400
300
200
100
0
Asia (Maximum Mycotoxin levels detected in ppb)
797
691
210
90
15
AFLB1 ZEA DON FB1 OTA
Multiple Mycotoxins are detected in raw material and finished feed samples
% Positive samples for Mycotoxins
Shi in
Contamination
100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0
Alfa B1
Zea T - 2 Fum B2 OTA
2001-2006 2006-2010 2011-2015
Year on year there is a shift in contamination from Aspergillus and Penicillium sp. to more of Fusarium sp. toxins
Luciano Pinotti et al
20 21

MYCOTOXINS
What is the safe limit?
European Food Safety Authority (EFSA) has provided
following recommendation for major mycotoxins for animal
feed
Mycotoxins
Aflatoxin B1
Ochratoxin
Citrinin
Maximum Permissible
Limits (EFSA*)
20 ppb
40 ppb
100 ppb
T2
200 ppb
Myth #2
THERE IS A SAFE LIMIT
TO MYCOTOXIN CONTAMINATION
Myth Buster
This is not true. Because even minimalistic
quantity of multiple mycotoxins can synergistically
have a deleterious effect on poultry health.
Zearalenone
400 ppb
DON
5000 ppb
*May cause toxicity at lower levels also if multiple Mycotoxins are present.
http://eur-lex.europa.eu/legalcontent/EN/TXT/PDF/?uri=CELEX:32006H0576&from=EN
22 23

MYCOTOXINS
How do they affect poultry
health and production?
Aflatoxin
Acute- High contamination: Gizzard
erosion, Hepatotoxic, Mortality in
severe cases, less feed intake, less
weight gain, reduced hatchability
T-2 toxin
Broiler- lesions in the digestive tract,
organs and skin, neural disturbances
and decreased weight gain.
Layer- reduction in egg production, and
hatchability% small mucosal ulcer in the
anterior portion of the gizzard
Ochratoxins
Broiler - High mortality, nephrotoxicity
and severely reduce weight gain
efficiency, lesions mostly seen after 3
weeks of age
Layers/ Breeders - Nephropathy,
reduced egg quality and production,
severe immunosuppression
Citrinin
Mainly nephrotoxic, reduced feed
consumption and body weight but
water consumption is increased
Myth #3
MYCOTOXINS CANNOT
CAUSE VACCINATION FAILURE
Zearalenone
Important for layers and
Breeders. Reproductive
system, Egg shell quality,
lower semen quality,
Deoxylivalenol
Less acute incidences.
Immunosuppression and Affects GIT,
Atrophy of liver, villus atrophy
Myth Buster
This is not true. Because immunosuppressive
Mycotoxins like Aflatoxin, Mycophenolic acid,
T-2 toxin, DON contamination can lead to significant
reduction in immune response.
Fumonisins
Effects seen after age of 2 weeks.
Body weight and feed intake
reduction, Liver enlargement and
damage, Kidney damage, reduction
of villus height which results in
production losses
Cyclopiazonic acid
High Mortality, Decreased weight gain,
nervous disorders and egg shell
abnormalities, Gizzard erosion
Mycophenolic acid
Immunosuppression
J.M. Milani PAUL E. NELSON et al Wageha Awad et al A.GENTLES et al Jewers K.
J.M. Milani
PAUL E. NELSON et al Wageha Awad
et al A.GENTLES et al Jewers K.
24 25

PESTICIDES
Toxicity and Effects on poultry
26 27

What are
Pesticides?
• According to WHO, Pesticides are chemical compounds that
are used to kill pests, including insects, rodents, fungi and
unwanted plants (weeds).
• Pesticides are used in public health to kill vectors of
disease, such as mosquitoes, and in agriculture, to
kill pests that damage crops.
Myth #3
Myth Buster
MYCOTOXIN IS THE ONLY
THREAT COMING FROM RAW MATERIALS
In addition to Mycotoxicoses, pesticide toxicity is also a ma er
of concern and can have similar impact as that of Mycotoxins
What is Pesticide
toxicity?
• Pesticides are potentially toxic to animals and humans,
and need to be used safely and disposed of properly
• When used irrationally these chemicals can enter into
feed and food chain via feed and food ingredients or by
leaching into ground water
• The Maximum residual limit has been set by European
commission which regulates the maximum amount of
pesticides residue that can enter feed or food chain.
• When residues are more than specified MRL values, they
may cause toxicity
http://www.who.int/topics/pesticides/en/
https://ec.europa.eu/food/plant/pesticides/max_residue_levels_en
28 29

How pesticides
enter bird’s body?
Pesticides
residue in crop
• Maize is the one of the most important ingredient in
animal feed.
• Contamination of maize with pesticide will allow entry
in feed chain
• Pesticides leeching in ground water will reach bird
through drinking water
Pesticides residues
leaching out
in ground water
Pesticides
residues
through feed
Storage grains
residue from
storage chemicals
How Pesticides
reach poultry feed?
Pesticides residue
through drinking
ground water
Residue accumulated
in muscles, fat tissues
& negative impact
on performance
Ref: Md. Wasim Aktar 2009, Vijay J. jadhav 2011
30 31

MYCOTOXINS
What is MRL-
Maximum
Residue Limit?
What is MRL for
Commonly Occurring
Pesticides in Maize?
The maximum residue limit (MRL) is defined as the
maximum concentration of a residue, resulting from
the registered use of a chemical, that is
recommended to be legally permitted or recognized
as acceptable in or on a food, animal feed. The
concentration is expressed in milligrams per
kilogram of the commodity.
Pesticide
Carbofuran
Deltamethrin
Atrazine
Malathion
Thiamethoxam
Thiram
MRL* (ppb)
50
1000
50
8000
50
100
*May cause toxicity at lower levels also if multiple mycotoxin infection is present
Ref: Md. Wasim Aktar 2009,
Vijay J. jadhav 2011
32 33

MYCOTOXINS
What is the
effect of
chemical
toxicity in
poultry?
Carbofuran
Use: During Storage and transportation of
Maize
Impact on Birds: Severe Kidney damage ,
Liver damage , Yellow colored bile, Subencapsular
haemorrhages in liver, severe
mortality up to 30% in acute cases
Thiram
Use: Seed Treatment of Maize before sowing
Impact on Birds: Liver enlargement & dysfunction,
hepatitis, degenerative changes focal necrosis
Clinical signs: anorexia, listless behaviour, dyspnea,
convulsions, and death due to cardiac arrest, inhibit egg
laying, soft egg shells, depressed growth and leg
abnormalities.
Deltamethrin
Use: Insecticide, during maize storage and
before sowing
Impact on Birds: Severe body weight drop,
poor hatchability, residue in the different organs
Malathion
Use: On field spray and while storage
Impact on Birds: Nervous disorders and
production stress
Thaimethoxam
Use: Insecticide
Impact on Birds: Severe liver damage,
growth suppression, impacts on hepatic
enzymes
Atrazine
Use: During Storage and trasnportation of Maize
Impact on Birds: Reproductive hormones, late
maturity, hepatoxicity
(Gupta 96), Hasegawa et al., 1988; Maita et al. 1991 Kaya & Bilgili,
1998 Lorgue et al., 1996 Guitart et al., 1996 Chandra et al 2013
Adejumo et al 2015 Wilhems 2006
34 35

MYCOTOXINS
Toxin Analysis methods
Thin Layer
Chromatography (TLC)
High Performance
Liquid Chromatography
(HPLC) with UV OR
Fluorescence(FD)
Enzyme Immunoassays
(EIA)
Liquid
Chromatography–
Mass Spectrometry
(LC-MS)
Gas
Chromatography–
Mass Spectrometry
(GC-MS)
LEAST SENSITIVE
MOST SENSITIVE
Maitree Suttajit Kabak B et al
36 37

PREVENTION
38 39

How can we
prevent
Mycotoxin
toxicity?
Myth #3
Myth Buster
TOXIN BINDERS CAN
ERADICATE ALL THE
TOXICITY
To minimize the impact of
toxicoses, an integrated
approach is required like
be er raw material
quality and storage
management
PRE HARVEST
• Using Fungi resistant
varieties of maize
• Controlling insect, pest
destruction of corn
• Harvesting crop at right
moisture
POST HARVEST
• Proper drying to reduce
moisture to 10-12%
• Proper storage of maize
- In Silos or raised
platforms
- In low temperature
- Away from damp
- Less humid environment
• Using fungi and insect
repellants in storage
IN FEED MILL
• Purchasing good quality of
maize (Moisture 11-13%)
• Optical sorters can be used
to sort the contaminated
seed
• Proper supplier profiling
• Redrying the doubted lot
• Physical separation of
contaminated seed
• Storage at feed mill is
equally important (same as
post harvest)
Maitree Suttajit Kabak B et al
40 41

MYCOTOXINS
Sampling: Critical Point
Ÿ
Quantifying Mycotoxin concentration of a maize lot based on
1 sample will give high uncertainty in results
-9
Mycotoxins are measured in nanogram (10 g) of ycotoxin
per gram of RM/Feed (parts per billion or ppb)
In a test procedure conducted by Whitaker et al shows
- Sampling variance of 5 kg sample of shelled
peanuts is 521.4
- Sample preparation variance is 59.2
- Analytical (TLC) variance is 50.1
The Variance in results can be reduced by obtaining a
representative sample correctly and using more sensitive
analytic methods
Obtaining a representative sample of feed or grains
Collect from different points from
- Moving stream of feed/grains
- From silos
- From truck
Ÿ M
Ÿ
Ÿ
Ÿ
Ÿ
1 ppb is equal to 1 second in 32 years or 1 drop of water
in Olympic size swimming pool!
Thomas. B. Whitaker, Raymond. D. Coker
42 43

MYCOTOXINS
Sampling: Critical Point
Collect incremental
samples of Feed/RM
from different lots
Mix all the
samples thoroughly
Take 300 g of
this sample
Send the sample
for Analysis
Maitree Suttajit Kabak B et al
44 45

Customer Lab
Services (CLS)
More than 1000 samples of Feed
and RM are analyzed for Mycotoxin
every month
Toxfin TM
Toxfin
Broad Spectrum
Toxin binders
• Toxfin
• Toxfin 360
0
Kemin’s 360
Mycotoxin Management
Technical
Services
A team of 24 on -
field Professional
Veterinarians
Kemin’s Knowledge Base
A dedicated research
and development team
to provide best
solutions
(Gupta 96), Hasegawa et al., 1988; Maita et al. 1991
Kaya & Bilgili, 1998 Lorgue et al., 1996 Guitart et al.,
1996 Chandra et al 2013 Adejumo et al 2015 Wilhems 2006
46 47