Impact of Mycotoxins on Reproductive Physiology.

1. FeedWorks Australia
Sept 14th – 18th 2022 FeedWorks Conference
Impact of mycotoxins on Reproductive Physiology

2. Mycotoxins: an overview
Field mycotoxin
(Fusarium)
Storage mycotoxin
(Aspergillus,
Penicillium)
Trichothecenes
Aflatoxins
Fumonisins
Ochratoxins
Deoxynivalenol
(DON)
T2 Toxin
Zearalenone
(ZEA)
Fumonisin B1
(FB1)
Aflatoxin B1
(AFB1)
Ochratoxin A
(OTA)
Barley
Rice
Corn
Sorghum
Wheat
Oat

3. zearalenone (ZEA) α-zearalenol (α-ZOL) ϐ-zearalenol (ϐ-ZOL)
zearalanone (ZAN) α-zearalanol (α-ZAL) ϐ-zearalenol (ϐ-ZAL)

4. Zearalenone
• Has a resorcylclic acid lactone structure and can cross cell membranes
• Binding to the cytosolic 17ß- estradiol (E2) receptors forming a
Zearalenone- E2R complex
• ZEA- E2R complex is transferred into the cell nucleus and binds to
specific nuclear receptors
• Responsible for mRNA synthesis besides E2
• ZEA interacts with both types of estrogen receptors
• ZEA is also a substrate for hydroxysteriod dehydrogenases which
create alpha-zearalenone and beta-zearalenone

5. Zearalenone, most common pathological
effects
• Anestrus
• Abortion
• Increased embryonic and fetal death
• Failure of induction programs with PGF2α
• Increased incidence of stillborn and splay legged pigs
• Gilts are more sensitive than sows
• Oocyte dies in the Graafian follicles despite signs of estrus there is no
ovulation

6. Zearalenone (ZEA)
• Rapidly absorbed
• 80-85% uptake
• Detected in the blood in about 30 min
• Entero-hepatic cycle prolongs retention of ZEA and deviations in the
blood stream retarding elimination and enhancing the duration of
adverse effects. ( D’Mello, J.P.F. et al., 1999)
• Swine most sensitive to Zearalenone, dairy cattle, chicken, turkey,
horses and sheep
• Heifers conception rates decline from 87% to 62%

7. Zearalenone
Vulva Swelling
Mammary
Enlargement

8. Zearalenone
Impact of ZEA and/or its
Metabolites on newborn piglet
and carries through the milk.

9. Zearalenone (ZEA) – Male responses
• Decrease testicular size
• Reduced libido
• Reduce body weight
• 30% weight reduction of testes
• Lowering testosterone
• Decreased fertility due to reduction in sperm quality and viability
• Apoptosis increases

10. Scrotal Insulation in the Boar
John Parrish,, University of Wisconsin

11. Citation Round Spermatids/
Sertoli Cell
% Loss
Theoretical 256 0
Franca et al., 2005 68 73.4%
Costa et al., 2013 21.5 91.6%
Parrish unpublished 30.4 88.1%
Losses During Spermatogenesis
(Boar)
Most losses (approx. 90%) occur during Mitotic processes
• A, I, B Spermatogonia or Primary Spermatocyte

12. Deoxynivalenol DON

13. Deoxynivalenol (DON/Vomitoxin)
 Reduced feed intake and weight gain (> 1 ppm)
 Diarrhoea, nutrients such as glucose, AA
 Reproductive failure
 Anorexia, PYY (gut satiety hormone), insulin
 Vomiting
 Death
 DON toxic effects on semen and impacts sperm in epididymis
 Presentation of analysis from samples from more than 100 countries suggest
that DON and ZEN are among the top mycotoxins 64% and 45% respectively.

14. T-2 Toxin

15. T-2 toxin impairs male reproductive function
• Down-regulated reproductive organs (testis, epididymis, and seminal
vesicle) weight, sperm concentration, sperm malformation
• Damage to ultrastructure of sperm and structure of testes
• Increase in reactive oxygen species (ROS)
• Decrease total anti-oxidation capacity (T-AOC)
• Limited superoxide dismutase activity in testes.
• World Health Organization listed T-2 toxin as unavoidable
contamination to human food. (Young et al., 2016)
• T-2 cytotoxic on rapidly dividing cells activates apoptosis

16. Fumonisins

17. Fumonisin (FB1)
0
20
40
60
80
100
120
containing mg/kg % sperm
Chart Title
diet 1 diet 2 diet 3 Control
5 mg/kg 10 mg/kg 15 mg/kg 0.2 mg/kg
83.3% 79.1% 59.6% 100%
Gbore, FA Animal 2009 Aug; 3(8): 1133-1137

18. Fumonisins
• Do not expose boars to FB1 higher than 5 mg/kg
• Toxicity due to enzyme mediated conversion of Sphingamine to
Sphingosine
• Increases SA/SO ratio interfering with cell cycle control and cell
function
• Poorly absorbed orally (3-6%) excreted rapidly in bile and urine
• FB1 cell signaling proteins protein kinase C (PKC)
• FB1 inhibits ceramide synthase pathway on the ascending aortic
impedance spectrum of pigs (Heart Failure)
• > 120 ppm fatalities will range 50-90%

19. Aflatoxins

20. Aflatoxins (AFB1)
• Decrease in body weight
• Poor feed conversion (FCR)
• Liver and kidney enzymes were severely elevated
• Testosterone level decreased
• Male birds pernicious effects of aflatoxins
• Poor libido, low semen motility, testicular atrophy
• Arrest of spermatogenesis

21. ELITOX®
Mycotoxin Eliminator

22. Immune response
• Zootechnical parameters
• BW, DWG, FI, FCR, egg production, reproductive performance,…
• Gut barrier functioning
• Tight junction proteins
• Gut morphology (villi, crypt,…)
• Microbiome in lumen
• Immune response
• Circulating immune cells (blood)
• Local immune responses (gut)
• Oxidative stress
• Endogenous enzymes (liver, gut, blood)

23. JAM1
JAM1
JAM1 JAM1
Claudin
Occludin
Tight Junction Complex
ZO1
ZO2
ZO3
ZO1
ZO2
ZO3
Actin cytoskeleton
Villi

24. Leaky Gut
Healthy Gut

25. Composition
• Enzymes
• Biopolymer
• Combination of minerals
• Natural extracts
• vitamins

26. Enzymes
• Selected enzymes must have the following characteristics:
• Stability during feed processing
• Efficacy under conditions of digestive tract (temperature, pH)
• Specificity towards the mycotoxin
• Products of degradaton or transformation not harmful to the
animals

27. Examples of deactivation:
OH
CH3
O
H
O
H
H
OH
1-(3,5-dihydroxyphenyl)-10’-
hydroxy-1’undecen-6’-one
lactonase
OH O
CH3
O
H
O
H
H
O
Zearalenone
Takahashi-Ando et al., 2002
oxidase
O
OH
O
OH
CH2OH
O
H
H
H
CH
3
H
H
DON
O
O
OH
CH2OH
O
H
H
CH
3
H
H
O
3-keto-DON
Shima et al., 1997

28. Biotransformation
• Conversions of mycotoxins into less or non-toxic molecules by
enzymes or microorganisms.
• Physical adsorption and this degradation takes place in the gastro-
intestinal tract.
• Astrid Koppenol, PhD, Ghent University of Belgium

29. Biopolymer
• Derived from prawn and crab shell
• Produced by deacetylation of chitin
• Biologically active compound
• Honeycomb like structure
• Anti-fungal and anti-bacterial
• Chitosan has antibacterial activity
• Supports active immune function of
enterocytes

30. Percentage of mycotoxins eliminated when including Elitox™
- summary of different gut simulator trials

31. Percentage of zearalenone disappeared when
including (ELITOX®) at 1.5 kg/MT of feed
100
200
300
82.00% 84.50%
80.70%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
%
Zearalenone
disappeared
Zearalenone level in feed (ppb)

32. Conclusions:
• 1,25 ppm resulted in significantly lower ADG and F/G
• Elitox restored ADG and F/G
• 2,5 ppm resulted in lower feed intake and
significantly lower ADG and F/G
• Elitox restored feed intake, ADG and F/G
• 1 kg/t proved to be the most cost-effective inclusion
rate
Trial: DON Challenge Piglets - USA

33. Conclusions
• A combination of ZEN and DON at levels in this trial
produced a dramatic reduction in health and
performance of challenged animals
• Visual signs of ZEN and DON challenge reduced by
~35% with 1 kg/t Elitox
• With 1.25 ppm DON and 750 ppb ZEN average
daily gain was improved by 31% and FCR improved
by 24% with 1 kg/t Elitox

34. A Report from the Regional Committees on Swine Nutrition and Management,
coordinated by Don Mahan, Ohio State University,Columbus, OH
Diet 1mg/kg DON 4 mg/kg DON
Control Biofix Plus Mycosorb Control Biofix Plus Mycosorb
Intake
(kg/d)
680 655 689 544 526 526
Gain (g/d) 462 449 467 340 362 336
FCR 1.48 1.46 1.49 1.60 1.54 1.64

35. Alkaloid Toxins
• Ergovaline from Perennial Rye grass Toxicosis
• Sorghum Ergot (Dihydroergosine... Structurally
and impact related to Ergovaline)
• Both Potent Vasoconstrictors- Big heat stress
impacts.
• Both Depress prolactin hormone , reducing
mammary development and yield of milk, and
also heat impacts -sweat

36. Recommendations
• Use 0.5kg/tonne of Elitox for lower risk grains (cost
$3/tonne) in pig grower finishers and poultry
• When the risk of mycotoxin is medium to high, use
1kg/tonne ($6/tonne)
• If presence of mycotoxin is guaranteed, then use 1.5
kg/tonne in pig breeder and weaner diets.