Aflatoxicosis

Introduction:-
Mycotoxins are naturally produced by Fungi .

( Metabolites of fungi ) .

Fungus ( singular ) & fungi ( plural )

Fungus is eukaryotic cells ( having characteristic of animal and plant's cell.)

Fungi => have cell membrane as well as cell wall.

Types of Fungi:-
i) Unicellular ( Yeast )

ii) Multicellular ( Mold => produce Toxins )

iii) intermediate ( having characteristics of unicellular as well Multicellular )

Fungi => reproduction => asexual and sexual.

Mycotoxins contamination :
i) Pre Harvest ( Before harvesting Crop attacked by fungus , e.g. Claviceps)

ii) Post Harvest ( After harvesting e.g. during cooking/feed manufacturing . E.g. Aflatoxins ( produce by
Aspergillus )

Aflatoxicosis
Aflatoxins are most potent hepatotoxin & hepato-carcinogenic ( Chronically ) .

( B¹ is the most toxic to liver and cause mutagenesis . WHO classified it as Class 1 ) .

• Alflaroxin Produce by Aspergillus flavus and parasitcus.

Note : Aflaxtoxin name derived from 1st letter of Aspergillus ( A ) and first 3 letters from Flavus (fla) =>
Afla - Toxin => Aflaxtoxin.

Other species :
A. Parasiticus , nominus , Fumigatus , Nidulans , Niger , terreus .

[ A. Flavus and Parasitcus most abundant in warm and hot areas of world. ]
Aflatoxins are divided as following:

B¹ , B² and G¹ , G² and M¹ , M² and Q¹ , Aflatoxicol.

• Toxicity order

AFB¹ > AFG¹> AFB²> AFG²

[ B² and G² are dihydoxy derivatives of B¹ and G² respectively]

[ Spores which gives Blue colour under fluorescent light=> Blue / B and which gives green colour =>
Green / G while ¹ and ² numbers indicate major and minor compounds] .

M¹ is metabolite of B¹ and M² is metabolite of B².

Note : [ M= Milk toxin => M¹ & M² secreted in milk of Ruminants after metabolism ] .

• Aflatoxicol and Q¹ are also metabolites of B¹.

[ B¹,B² and M¹,M² => have Difurocoumarocyclopentenone while G¹,G² and Q¹ => Difurocoumarolactone ]

• Host Range
Note : Aflatoxins affect wide range of animal's species. ( All birds are susceptible

Species. But duck and turkey birds are more sensitive bcz their microsomal enzymes in liver ( abundant )
Cyp450 which involve in AB¹ biotransformation ) .

• Note : Similarly younger birds are more susceptible than older bcz of microsomal enzymes in liver are
more active towards AB¹ biotransformation than older .

Factors:
i) contaminated Litter ( during harvesting)

ii) Contaminated Feed ( Preharvest/ Postharvest)

iii) Poor ventilation ( => increase Humidity=> sporulation => toxins )

iv) High temperature ( => sporulation => toxins )

v) Immunocompromised birds
Vertical transmission:
• Aflatoxins affect uterus/ hatchery contamination => egg shell contamination by Aspergillus spores
chick pipping during hatching => inhale spore => Aspergillosis .

( piping chick break egg shell during hatching. ).

Pathogenesis:-
1- Inhalation route: Aspergillus Spores inhale => if Spores are of A. Flavus ( 4-6 um ) affect upper
respiratory tract while Spores are A. Fumigatus (2-3 um ) affect lower respiratory tract.

Spores => vegetate => produce toxins => upper respiratory tract (A.flavus ) / lower respiratory tract ( A.
Fumigatus ) => irritate Mucosa => increase Vascular permeability in Lamina propria => plasma oozing out
in lumen and spores also direct stimulate sensory receptors=> increase vagal tone => increase Mucus
production( by goblet cells ) in Tracheal, bronchial & parabronchial lumen ( Trachea= Upper respiratory
tract & Bronchi & parabronchi = Lower respiratory tract) =>increase thick Mucus production =>
cilliostasis => Mucoid plug => Gasping ( bcz of narrow lumen ) => Asphyxia=> Death .

2- Ingestion Route => Gut => irritate GiT mucosa => increase permeability of vasculature => decrease
nutrients absorption ( specially decrease iron absorption , Vit A , D³ , ca² and other nutrients) => loose
droppings ( osmotic diarrhea ) .

Toxins liberated by Spores also absorb from Gut ( Toxins are highly lipophilic and easily cross cell
membrane. ) => liver [ in liver AB¹ converted into AM¹ , AFBO ( Aflaxtoxin Exo 8,9 Epoxide & Aflaxtoxin
Endo 8,9 Epoxide ) , Aflatoxicol and ABQ . ]

( Toxins also convert into toxic metabolites in GuT )

AFBO => bind with DNA ( at N⁷ position guanine and Formed=>8,9-dihy-dro-8-(N7-guanyl)-9-

hydroxy-AFB1 ) . AFBO also binds to other cellular proteins and molecules. ) . This DNA-AFBO metabolite
adducts cause => Toxicity and due to Damage to DNA => initiates carcinogenic proliferation .

AFBO also cause dysfunction of GST ( Glutathione S transferase ) which normally involve in it's
detoxification.

AFBO transport to other organs of body via circulation => AFBO-DNA adduct cause mutagenesis.

[ Mutagenesis=> DNA-AFBO adduct ( 8,9-dihy-dro-8-(N7-guanyl)-9-hydroxy-AFB1) = formed guanine ( G

) base ( which purine ) and bind to Thymidine ( T ) which is pyrimidine and leads to transversion
mutation. This mutation affect p53 gene( tumour suppressor gene ) .
 Note : Transversion mutation => changing of nucleotide base from purine to pyrimidine or pyrimidine to
purine. As DNA-AFBO do transversion mutation ( Change Guanine nucleotide base with Thymidine . As
we know Guanine( G ) nucleotide base attached with Cytosine ( G ) . So when G is changed to T then
base pair on leading strand then next base on lagging strand ( alternative strand ) will be Adenine ( A ) .
That's how whole genome will disturb => gene mutation => Disturb length of telomeres => change
checkpoints of cell cycle = Tumours. ]

[ normally => Endo and exo peroxides of AB¹ is bind with Glutathione and convert to nontoxic
metabolite by enzyme Glutathione S transferase and formed AFB-mercapturate ( nontoxic ) => excrete .
But higher concentration of ABFO leads to dysfunction of GST ( as mentioned above ). ]

Immunosuppresion: AFBO => affect directly bone marrow=> decrease lymphocytes , Erythroblasts and
also decrease phagocytic activity of Macrophages ( also decrease cytokines production in Chronic
condition => decrease antibodies production => immunosuppresion ( bird more susceptible to
secondary viral / bacterial / parasitic disease )

Clinical Signs:
• General
i) Anemia cause paleness of wattles , comb , bone marrow ( because of iron deficiency & damgaed to
bone marrow decrease in Erythroblasts explained in Pathogenesis)

( microcytic hypochromic Anemia ) .

ii) Increase thirst ( because of loose droppings)

iii) Pyrexia (in acute case due to IL-6 , IL-1 , TNF_x which are released in body => damage caused by
Toxins.)

[ acute Aflatoxicosis when large concentration of toxins ingested while in chronic upto minimum one
week ]

iv ) Hemorrhages under skin ( bcz toxins cause decrease in Factor X , VII and IX and prothrombin =>
increase clothing time and also cause increase in Vascular fragility => Hemorrhages .)

v ) Decrease Production

vi) Decrease hatchabilty & increase embryonic mortality and malformations ( enlarged eye ball and liver
)

vi) Depigmentation ( bcz decrease in beta carotene )

• Respiratory
i) Open mouth breathing => Gasping & Dyspnea ( bcz of excessive Mucus in lumen => narrow air
passage => decrease oxygenation. )

[ Gasping=> without noise while in other Respiratory diseases like IB Gasping with noise) .

ii) Nasal discharge

iii) Cyanosis ( Bluish discolouration of skin , comb and wattles bcz of decrease oxygenation => which due
to Dyspnea. )

• Digestive
i) Loose & foetid droppings ( Explained above in Pathogenesis)

• CNS
i) Neck twisting [ Toxins damage to cortex => convulsions ( Neck twisting if brain stem damaged ) .]

ii) Blindness (toxins absorb from parabronchi cause infection=> eye . )

iii) Paralysis & Opisthotonos ( AFBO => enters in neurons=> decrease Acetylcholine production and
increase Acetylcholinesterase concentration => paralysis.)

Postmortem lesions:
i) Congested and edematous carcass ( bcz of increase fragility of vasculature )

ii) Swollen , Congested or yellowish brown ( fatty ) liver also may be there necrotic foci ( black dots on
liver surface )

Liver will be harder & pale in Chronic form ( not fragile )

iii) Tumors on liver surface ( explained in Pathogenesis)

iv) swollen Kidneys and hemorrhagic

v ) May be there are Hydro pericardium and ascites ( bcz increase in fragility and congested liver )
 Diagnosis:
i) History

ii) Postmortem lesions

iii) Aflatoxins detection ( Fluorescence under UV light )

iv) Feed testing

Differential Diagnosis : -
Aflatoxicosis cause immunosuppression leads to secondary viral / bacterial / parasitic ( Emeria protozoal
) infection.

Management: -
i) Proper storage of Feed

ii) Control of insects and rats

iii) Proper Ventilation ( avoid sporulation in litter / feed by decreasing Humidity in shed )

Treatment:-
i) Change Feed source

ii) Mycotoxin Binder

iii) Vit E and Se ( decrease oxidative stress ) + ViT A , C and bcomplex+ Electrolytes

( Electrolytes=> loose droppings

Fat soluble vitamins provide exogenously bcz of bile duct and liver damaged and loose droppings)

iv) Cynara extract ( Antioxidant and anti-tumors)

Mycotoxin Binder:-
A) Aluminosilicates :
Clay based ( Zeolite , clinoptilolite , kaolinite, HSCAS )

• HSCAS = Hydrated Sodium calcium Aluminosilicates

• Never use clay based mycotoxin binders above 2% in diet .

( Theseay cause decrease in nutrients absorption).

B ) Yeast based
i) Mannan ii) Mannan oligosaccharides (MOS)

iii) Saccharomyces cerevisiae ( bind with toxins and convert them into nontoxic metabolite then excrete
through feces )

C ) Probiotics based:
Enterococcus , lactobacilli , streptococci ( some species are beneficial ) bind with toxins and convert
them into nontoxic metabolite then excrete through feces . Probiotics & Yeast also stimulate immune
system and increase nutrients absorption.

D) polymer polyvinylpyrrolidone (PVP) :

Bind with toxins

Note:
Copper sulphate & antifungal have antifungal activity ( don't bind with mycotoxins ) .

Yeast based , Probiotics , HSCAS , Clinoptilolite => effective against Aflatoxins.

[ Can be use in combination => yeast + probiotics

Probiotics and Yeast bazed => improve nutrients absorption as well as act as toxin binder ] .
Regards;
Dr Hamza Mohsin ( DVM BZU)

Dr Mudassar Ahmad ( DVM Riphah international University Lahore )

Dr Awais Mohsin ( DVM BZU)

Dr Beenish Abbas ( DVM BZU )

Dr Saira Muzammil ( DVM UVAS)

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