Jai shree ram
12/11/2013

2
University of Horticultural Sciences, Bagalkot
Kittur Rani Channamma College of Horticulture, Arabhavi

MYCOTOXINS AND THE...
12/11/2013

4
What are Mycotoxins?
Mycus = Fungus / Mold
Toxicum = Poision
Mycotoxins are toxic substances / secondary
metabolites produ...
Mycotoxicosis is used to describe the action
.
characterized as:
Acute mycotoxicosis- high dose and rapid
Chronic mycotoxi...
History
Turkey X Disease
Death of about 100,000 turkey poultry birds ---- United Kingdom in
1960 ----ingestion of a poultr...
1093 in France - holy fire – - St. Anthony’s fire Ergot of rye - Claviceps purpurea - Ergot alkaloids Russia in 1926, Irel...
Cont…

• Alimentary Toxic Aleukia in Russia during World War 2 (T-2)
death of > 50000 soldiers
• 1930 ---- Moldy rice in J...
Why are they important?
• Destroy crops and causes economic loss
• Ingested through contaminated food
• Cancer, birth defe...
Where do they come from ?
 Moisture in crops
 Contamination during




Handling
Storage
Processing of foods

 Fungi ...
Mycotoxin and climate change
Extreme precipitation, storms and
floods
Cause moist conditions

Drought
Weakens seed kernels...
Determination of mycotoxins
Presumptive tests

Biological assays

Chemical analysis
HPLC- MS Test

(Miraglia et al., 2009)
Fig 1:- ELISA kit for the determination of mycotoxins. Hexagon-triangles
enzyme- mycotoxin conjugate, triangles free mycot...
Examination of Fungal Colonies under UV
Light

Fig 2 :- Two non-aflatoxigenic (top colonies) and one aflatoxigenic (bottom...
Lateral Flow Immunochromatography

(Vishay et al., 2009)
12/11/2013

17
Groups of mycotoxins
Aflatoxins
Ergot alkoloids
Ochratoxins
Patulin
Fusarium toxins
Zearalenone
(Abid et al., 2003)
12/11/...
Mycotoxin distribution
Mycotoxin

Fruits,
vegetables

Cereals

Milk and
milk
products

Coffee,
wine

Nuts,
spices

Baby
fo...
20

(Miraglia et al., 2009)
Producing:
Aspergillus flavus and A parasiticus

Important metabolite is Aflatoxin B1, B2, G1, G2, M1
Warm temperature con...
Conditions favorable for Aspergillus
— High moisture
— Drought stress
— Insect damage

Aspergillus Growth on Corn

Aspergi...
A. fumigatus

A. nidulus

A. niger

A. oryzae

A. clavatus

A. flavus
Chemical structure of aflatoxins
B

A

(A) The B-type aflatoxins are characterized
by a cyclopentane E-ring. These
compoun...
AF binds to DNA

Aflatoxin B1 binds to
DNA at the guanine
base in liver cells,
corrupting the genetic
code that regulates ...
Table 1:-Acceptable levels of Aflatoxins in Food & Feeds
(Source: www.fda.gov)
Action Level

Commodity

Species

0.5 ppb A...
Aflatoxins contamination in chilli samples from Pakistan
R. Russell and M. Paterson
April 2006, Pakistan

12/11/2013

27
Table 2: Description of the chilli samples used in this study
(Russell and Paterson, 2007, Pakistan)
Sample
number

Locati...
Aflatoxin Contamination of Red Chili Pepper From Bolivia and
Peru, Countries with High Gallbladder Cancer Incidence Rates
...
Table 3: Concentrations and Recovery Rates of Aflaoxins B1, B2, G1,
and G2
(Takao et al., 2012 , Peru)
Red chili peppers

...
Fig 4 : HPLC Chromatograms of Each 10 μg/Kg Portion of
Aflatoxin Standard (A) Bolivian Red Chili Pepper (B).Aflatoxins B1 ...
Effects of Aflatoxin B1 (AFB1) on Liver of chickens
(Severe periportal cytoplasmic vacuolation of hepatocytes)

Control
12...
Aflatoxin
12/11/2013

Control
(Hsieh, 2009, Lowa)
Tumor induction in fishes

(Santis, 2008)

12/11/2013

34
Effects of aflatoxin on pigmentation
(Doerr et al., 1993)

12/11/2013

(Aflatoxin +)

(Aflatoxin -)

35
Produced by –Penicillium and Aspergillus

Growing in different climates and on different plants - contamination of food cr...
Structure of ochratoxin

12/11/2013

37
(Aish et al., 2000, U.K.)
Effects of OA on Pigmentation

OA +

OA -

12/11/2013

(Turner et al., 2007)
12/11/2013

39
(Phillips et al., 2007, U.S.A.)
The fuminosins ---- due to outbreak of equine leukoencephalomalacia in South Africa in
1970
Produced by > 11 species of th...
SPHINGOLIPIDS


Found in membranes, brain, nerves



Functions include:
• Internalization of lipids & proteins
• Cell-ce...
Mechanism of Toxic Action of FB1 - disruption of sphingolipid
biosynthesis
(Broomhead et al., 2002, USA)

Palmitoyl-CoA + ...
Effect of FB1 on ducklings

Control
(3.5 kg wt.)
12/11/2013

400 µg/kg
FB- 1 toxin injected
(900mg wt.)
(Musser and Platte...
Fig 6:- Fuminosin Induced Equine Leucoencephalomalacia
(Diaz et al., 2000)
12/11/2013

44
(Leeson et al., 1995)

Produced by Trichothicium, Fusarium
tricinctum (T-2 toxins),
Fusarium
graminearum (vomitoxin or DON...
Control (5.6 kg )

T-2 toxin 550µg/kg affected (1.9kg)

Fig 7 :- Effects of T-2 on Growth of the chickens
(Leeson et al., ...
Fig 8 :- Oral Lesions on chickens caused by T-2
toxin
(Leeson et al., 1995)
12/11/2013

47
The 40 ergot alkaloids -Claviceps purpurea can be divided into three groups:
Lysergic acid (e.g. ergotamine and ergocristi...
 Indole
NH

 Tryptamine

NH2
CH2

CH2

NH

NH2

 Tryptophan

CH2
NH

CH
COOH

(Swamy and Devegowda, 1998)
Zearalenone
• First described in US 1920s
• F-2 toxin
• Fusarium roseum,
F. tricinctum, F. gibbosum,
F. oxysporum, F. moni...
Phenolic compound
Zearanol (zearalenol)
Moisture - > 23%
Temp – 12 - 25 C,
Affects on reproductory system – hormonal imbal...
Effects of ZEN on humans
(Sundlof and

Dose
1 µg/day
6 µg/day
12.6 µg/day

Strickland, 1986)

Clinical Lesions
Vulva swell...
Co- occurrence of aflatoxins, ochratoxin A and zearalenone in
Capsicum powder samples available in the Spanish market
L. S...
Fig 9 : Aflatoxins, ochratoxin A and zearalenone distribution in Capsicum
powder samples
(Santos et al., 2010, Spain)
12/1...
Patulin
Patulin is produced by Penicillium clariform, P. expansum, P.
patulum and by Aspergillus spp
Sources :- Bread, sau...
Table 6: Effect of chronic oral administration of apple juice contaminated with
patulin (152.5 µg/kg) on dopamine (DA) con...
SCREENING OF MYCOTOXINS IN FRUITS AND
VEGETABLES GROWN IN SINDH, PAKISTAN
Najmus Sahar, Mubarik Ahmed, Zahida Parveen, Amb...
Table 7: Qualitative analysis of Aflatoxin in fruits and vegetables samples by TLC method
(Najmus 2009, karachi)
S. No.

N...
Table 8:- Aflatoxin contamination in medicinally important dried
rhizomes samples
(Anju and Geeta, 2008, Jammu)

Plants

S...
Ecological aspects of distribution of potential
toxin-producing micromycetes on stored apple fruit
(Alma and Elena, 2006, ...
Table 9: Distribution of fungi on healthy and rotten apple fruits
(Alma and Elena, 2006, Lithuania)
Fungal species
Distrib...
Cont…
Penicillium corylophilum

0

25

Penicillium clavigerum

0

8

Penicillium corymbiferum

0

10

Penicillium expansum...
Table 10: Potentially toxin-producing micromycete species isolated from store house
environment.
(Alma and Elena, 2006, Li...
Mycotoxigenic fungi and mycotoxins in fast food of Bihar

H. K. Chourasia, Sanjay Kumar Suman and Pramila Prasad
Journal o...
Table 11:-Incidence of fungi (%) in raw materials of fast foods of Bihar
(Chourasia et al., 2008, Bhagalpur)
Raw
materials...
Fig 11 : Associaion of microbes with raw materials of fast foods
a: Cabbage, b: carrot, c: Beet, d: egg, e: noodles, f:piz...
12/11/2013

67
12/11/2013

68
Effects of Mycotoxins
ZON
• Irregular heats
• Low conception rates
• Ovarian cysts
• Embryonic Loss

T2-Toxin, DON, AFB1
•...
Mycotoxicoses in dairy
T2-Toxin: dermal lesions

Zearalenone

Trichothecenes: Ketosis
Swollen vulva, Uterus prolapse

Deox...
Review of genetic studies of susceptibility to facial eczema
in sheep and dairy cattle
C. A. Morrisa, S. H. Phuab, N. G. C...
Fig12 :- Mean loge[GGT] of sheep groups, after dosing with the FE toxin sporidesmin and classified
By the progeny-test sta...
Fig 13 :- Mean loge[GGT] of calf groups, sired by Holstein Friesian bulls that had been
scored (performance tested) as wea...
Carry-over
metabolites

Poor
performance

Uterine
infections

Economic
Impact

Higher
Incidence
Diseases

Cystic ovaries

...
Impact of
mycotoxins in milk quality

12/11/2013

75
Carry-over of aflatoxins in dairy
Aflatoxin B1 is carried over into the milk as
Aflatoxin M1
From cow…

Human health conce...
Fig 14:- Effect of mycotoxin binder on Daily Milk production at
the Jersey Farm in Bhopal.
(Whitlow, 1986, Bhopal)
12/11/2...
 Neural tube birth defects
 Esophageal cancer
 Hepatocellular carcinoma

 Carcinogenesis
(Amer, 2009)
12/11/2013
Indigenous population that consumes high amounts of maize
as their staple food
Incidence of neural tube defects
(NTD per 1...
Fabio Galvano, Amedeo Pietri, Terenzio Bertuzzi, Luigi
Gagliardi, Sabina Ciotti, Stefano Luisi, Matteo Bognanno, Luca La
F...
Fig 16 : Chromatogram (HPLC; fluorescence detection) of: a) an OTA standard
solution (479 ng/L), equivalent to 47.9 pg of ...
Fig 17 : Chromatogram (HPLC; fluorescence detection) of: a) an AFM1
standard solution (62 ng/L), equivalent to 6.2 pg of A...
Anencephaly

Spina Bifida

Encephalocele

Fig 18 :- Examples of suspected fumonisin-associated birth defects: anencephaly,...
Spina bifida (a common form of NTD)
Spina bifida (Latin: "split spine") is a developmental birth defect caused by the
inco...
Encephalocele: which results when
other parts of the brain remain
unfused.

Anencephaly : a condition in which the
portion...
LUNG TUMORS
Adenocarcinoma
Large Cell
Aflatoxin Positive

Mesothelioma
Ochratoxin found in
chest wall, diaphragm.
No ochra...
Removal or Elimination of
Mycotoxins

• Pre harvest control
• Resistant varieties
• Crop rotation
Inactivation of Mycotoxi...
Conclusion............!!!!!
• Mycotoxins are highly stable compounds and ubiquitous
• Causes severe crop loss because cont...
Mycotoxin ppt by archana m. madalagi.
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  • Jai shree ram
  • Powdery mildew of Rose and Its Management Archana A. Madalgi
  • INTRODUCTIONHISTORYGROUPS OF MYCOTOXINSEFFECT OF MYCOTOXINS ON PLANTSEFFECT OF MYCOTOXINS ON ANIMALSEFFECT OF MYCOTOXINS ON HUMANSMANAGEMENT OF MYCOTOXINSCONCLUSION
  • (Agrios, 2000)
  • Lima et al., 2009
  • HPLC- MS Test
  • Distribution of mycotoxins
  • Conditions favorable for aspergillus
  • G- ground chilli P- chilli pods
  • (Santis, 2008)
  • Co- occurance of aflatoxins, ochratoxin A and zearalenone in Capsicum powder samples available on the Spanish market L. Santos, S. Marin, V. Sanchis and A.J. Ramos Spain 2010
  • Anju and geeta, 2008, Jammu
  • Alma and elena, 2006, Lithuania.
  • Cont…
  • H. K. Chourasia, Sanjay KumarSuman and Pramila Prasad J. mycol pl pathol, vol.38, No.3 2008
  • Table:- incidence of fungi (%) in raw materials of fast foods of bihar
  • Effect of 350 ppb Dietary T-2 Toxin on Daily Milk production at the Randleigh Jersey Farm Whitlow, 1986, NCSUBinder is Clay carbon
  • Mycotoxin ppt by archana m. madalagi.

    1. 1. Jai shree ram
    2. 2. 12/11/2013 2
    3. 3. University of Horticultural Sciences, Bagalkot Kittur Rani Channamma College of Horticulture, Arabhavi MYCOTOXINS AND THEIR EFFECTS ON PLANTS AND ANIMALS By: Archana A. Madalgi UHS12PGM223 12/11/2013 3
    4. 4. 12/11/2013 4
    5. 5. What are Mycotoxins? Mycus = Fungus / Mold Toxicum = Poision Mycotoxins are toxic substances / secondary metabolites produced by fungi (molds) growing on crops in the field and in storage. (Phreusa, 2009) 12/11/2013 5
    6. 6. Mycotoxicosis is used to describe the action . characterized as: Acute mycotoxicosis- high dose and rapid Chronic mycotoxicosis- slow development of infection (Harvey et al., 1991) Toxicoses – injection of toxic secondary metabolites (Osborne et al., 1982) Mycosis - generalized invasion of living tissue (McKinney et al., 1973) Mycetism - poisoning of organisms (Phillips et al., 2007) 12/11/2013 6
    7. 7. History Turkey X Disease Death of about 100,000 turkey poultry birds ---- United Kingdom in 1960 ----ingestion of a poultry feed containing Brazilian groundnut cake---- AFLATOXIN Chemical and microbiological investigations ---- toxic effects produced by the Brazilian groundnut cake ---- presence of quantities of four secondary metabolites ---- Aspergillus flavus in the diet (Agrios, 2000) 7 12/11/2013
    8. 8. 1093 in France - holy fire – - St. Anthony’s fire Ergot of rye - Claviceps purpurea - Ergot alkaloids Russia in 1926, Ireland in 1929, France in 1953, India in 1958, and Ethiopia in 1973 8
    9. 9. Cont… • Alimentary Toxic Aleukia in Russia during World War 2 (T-2) death of > 50000 soldiers • 1930 ---- Moldy rice in Japan - > 15000 people suffered • 1972 in China – 800 persons affected due to consumption of moldy sugar cane • Gibberella ear rot – contaminated beer– vomitoxin and zearalenone – cattle's, swine, chickens, horses, dog , fishes and humans in Canada • Fusarium toxins - in Virginia – Crazy horse disease- swelling of respiratory system in - human cancers • Dog Food- burning and death - Aflatoxin Poisoning 2006 in Kenya (Agrios, 2000) 12/11/2013 9
    10. 10. Why are they important? • Destroy crops and causes economic loss • Ingested through contaminated food • Cancer, birth defects, liver damage, and nervous tissue damage, etc. • Not destroyed in normal cooking procedures. • Played an important role in some historical events • Annual losses - $5 billion (Paterson et al., 2011) 12/11/2013 10
    11. 11. Where do they come from ?  Moisture in crops  Contamination during    Handling Storage Processing of foods  Fungi present in air, water, food and dust (Lima et al., 2009) 12/11/2013 11
    12. 12. Mycotoxin and climate change Extreme precipitation, storms and floods Cause moist conditions Drought Weakens seed kernels of plants, allowing greater fungal contamination Increased temperatures Promote fungal growth (WHO, 2009) 12
    13. 13. Determination of mycotoxins Presumptive tests Biological assays Chemical analysis
    14. 14. HPLC- MS Test (Miraglia et al., 2009)
    15. 15. Fig 1:- ELISA kit for the determination of mycotoxins. Hexagon-triangles enzyme- mycotoxin conjugate, triangles free mycotoxin, Y- anti mycotoxin (Varma et al., 2010) antibody. 15
    16. 16. Examination of Fungal Colonies under UV Light Fig 2 :- Two non-aflatoxigenic (top colonies) and one aflatoxigenic (bottom colony) strains of parasiticus visualized (a) under visible light; (b) 365 nm UV light. The ring around the aflatoxigenic strain displays blue fluorescence; (c) room temperature phosphorescence was photographed with a digital camera with a 2.5mp exposure after switching-off the UV lamp. (Rojas et al., 2007)
    17. 17. Lateral Flow Immunochromatography (Vishay et al., 2009) 12/11/2013 17
    18. 18. Groups of mycotoxins Aflatoxins Ergot alkoloids Ochratoxins Patulin Fusarium toxins Zearalenone (Abid et al., 2003) 12/11/2013 18
    19. 19. Mycotoxin distribution Mycotoxin Fruits, vegetables Cereals Milk and milk products Coffee, wine Nuts, spices Baby food Aflatoxins B1B2G1G2 Aflatoxin M1 Trichothecenes (DON, T2 toxins) Zearalenone OTA Patulin (Kofi, 2009) 12/11/2013 19
    20. 20. 20 (Miraglia et al., 2009)
    21. 21. Producing: Aspergillus flavus and A parasiticus Important metabolite is Aflatoxin B1, B2, G1, G2, M1 Warm temperature conditions favours the development Potential sources tomato, chilli, cocoa, coffee, groundnut, soybean, corn, milk and milk products, meat etc.... Effects: Inhibit protein synthesis Tumour cell development cancer cause mutation cell toxicity liver damage (Williams et al., 2004)
    22. 22. Conditions favorable for Aspergillus — High moisture — Drought stress — Insect damage Aspergillus Growth on Corn Aspergillus parasiticus Aflatoxin B1 12/11/2013 22 (Williams et al., 2004)
    23. 23. A. fumigatus A. nidulus A. niger A. oryzae A. clavatus A. flavus
    24. 24. Chemical structure of aflatoxins B A (A) The B-type aflatoxins are characterized by a cyclopentane E-ring. These compounds have a blue fluorescence under long-wavelength ultraviolet light. R3 R1 R4 (B) The G-type aflatoxins, with a green fluorescence, have a xanthone ring in place of the cyclopentane. (C) Aflatoxins of the B2 and G2 type have a saturated bis-furanyl ring. Only the bis-furan is shown. R1 R2 C D R1 R1 (D) Aflatoxin of the B1a and G1a type have a hydrated bis-furanyl structure. (Abid et al., 2003) 12/11/2013
    25. 25. AF binds to DNA Aflatoxin B1 binds to DNA at the guanine base in liver cells, corrupting the genetic code that regulates cell growth. Out- of- control cells grow into tumors that eventually become cancerous. 25 (Hsieh, 2009, Lowa)
    26. 26. Table 1:-Acceptable levels of Aflatoxins in Food & Feeds (Source: www.fda.gov) Action Level Commodity Species 0.5 ppb Afla M1 Milk Humans 20 ppb Any food Humans 20 ppb Feed All species of animals Exceptions: 300 ppb 300 ppb Cotton seed Meal used in Feed All species of animals Corn Beef Cattles 200 ppb Corn Swine 100 ppb Corn 12/11/2013 Breeding Cattle, breeding swine, and mature poultry 26
    27. 27. Aflatoxins contamination in chilli samples from Pakistan R. Russell and M. Paterson April 2006, Pakistan 12/11/2013 27
    28. 28. Table 2: Description of the chilli samples used in this study (Russell and Paterson, 2007, Pakistan) Sample number Location Aspergillus flavus (cfu) Total aflatoxin (µg/kg) 1 G Karachi 0 96.2 2 G Islamabad 0 66.1 3 G Islamabad 103 61.3 4 G Islamabad 103 48.9 5 G Faislabad 102 46.8 6 G Islamabad 104 40.1 7 G Lahore 103 34.3 8 G Lahore 2 102 25.5 9 G Faislabad 9 102 6.8 10 P Peshwar 2 102 6.6 11 P Karachi 6 102 1.7 12 P Lahore 2 101 0.2 13 12/11/2013 Sample P Lahore 2 100 0.1 G- Ground chilli P- chilli pods 28
    29. 29. Aflatoxin Contamination of Red Chili Pepper From Bolivia and Peru, Countries with High Gallbladder Cancer Incidence Rates Takao Asai, Yasuo Tsuchiya, Kiyoshi Okan, Alejandro Piscoya, Carlos Yoshito Nishi, Toshikazu Ikoma, Tomizo Oyama, Kikuo Ikegami, Masaharu Yamamoto Asian Pacific Journal of Cancer Prevention, Vol.13, Peru, 2012 12/11/2013 29
    30. 30. Table 3: Concentrations and Recovery Rates of Aflaoxins B1, B2, G1, and G2 (Takao et al., 2012 , Peru) Red chili peppers Aflatoxins (μg/Kg) B1 B2 G1 G2 Bolivia (1) N.D. N.D. N.D. N.D. (2) N.D. N.D. N.D. N.D. (3) 11.3 0.6 N.D. N.D. Peru (1) 0.9 N.D. N.D. N.D. (2) N.D. N.D. N.D. N.D. (3) N.D. N.D. N.D. N.D. 0.7 N.D. N.D. N.D. (2) N.D. N.D. N.D. N.D. (3) 2.3 N.D. N.D. N.D. Japan (1) N.D. N.D. N.D. N.D. Recovery rate (%) 102.1 101.6 103.6 109.1 China (1) N.D.:- Not detected 12/11/2013 30
    31. 31. Fig 4 : HPLC Chromatograms of Each 10 μg/Kg Portion of Aflatoxin Standard (A) Bolivian Red Chili Pepper (B).Aflatoxins B1 and B2 were detected at concentrations of 11.3 and 0.6 μg/Kg, respectively. 12/11/2013 31 (Takao et al., 2012 , Peru)
    32. 32. Effects of Aflatoxin B1 (AFB1) on Liver of chickens (Severe periportal cytoplasmic vacuolation of hepatocytes) Control 12/11/2013 AFB1 (Hsieh, 2009, Lowa)
    33. 33. Aflatoxin 12/11/2013 Control (Hsieh, 2009, Lowa)
    34. 34. Tumor induction in fishes (Santis, 2008) 12/11/2013 34
    35. 35. Effects of aflatoxin on pigmentation (Doerr et al., 1993) 12/11/2013 (Aflatoxin +) (Aflatoxin -) 35
    36. 36. Produced by –Penicillium and Aspergillus Growing in different climates and on different plants - contamination of food crops Outbreaks of Balkan nephropathy, a fatal chronic renal disease (Petkova and Castegnarom, 1985) Found in a wide range of human foods such as cereals, beer, wine, cocoa, coffee, dried vine fruit, spices and in meat products. (Abdel et al., 1999) Mode of action :- deformation of cells by producing phenyl - alanine In food is widespread and shown nephrotoxic, hepatotoxic, and carcinogenic (Aish et al., 2000, U.K.) 12/11/2013 36
    37. 37. Structure of ochratoxin 12/11/2013 37 (Aish et al., 2000, U.K.)
    38. 38. Effects of OA on Pigmentation OA + OA - 12/11/2013 (Turner et al., 2007)
    39. 39. 12/11/2013 39 (Phillips et al., 2007, U.S.A.)
    40. 40. The fuminosins ---- due to outbreak of equine leukoencephalomalacia in South Africa in 1970 Produced by > 11 species of the fungus Fusarium ( F. roseum, F. oxysporum, F. ricinctum F. graminearum etc..) Divided into structurally distinct groups, four of which have been designated A, B, C and P fuminosins Disrupt sphingo lipid metabolism FB1 implicated in neural tube defects in Texas population consuming corn (1989 – 1991) Moisture >23%; Temp – 28ºC Water soluble Esophageal cancer (Marasas, 2001) 12/11/2013 40
    41. 41. SPHINGOLIPIDS  Found in membranes, brain, nerves  Functions include: • Internalization of lipids & proteins • Cell-cell communication • Regulation of cell surface receptors • Regulation of ion pumps (Rheeder et al., 2002) 12/11/2013 41
    42. 42. Mechanism of Toxic Action of FB1 - disruption of sphingolipid biosynthesis (Broomhead et al., 2002, USA) Palmitoyl-CoA + Serine Sphinganine FB1 acyltransferase Ceramide Sphingosine 12/11/2013 Sphingolipids turnover 42
    43. 43. Effect of FB1 on ducklings Control (3.5 kg wt.) 12/11/2013 400 µg/kg FB- 1 toxin injected (900mg wt.) (Musser and Plattener, 1997)
    44. 44. Fig 6:- Fuminosin Induced Equine Leucoencephalomalacia (Diaz et al., 2000) 12/11/2013 44
    45. 45. (Leeson et al., 1995) Produced by Trichothicium, Fusarium tricinctum (T-2 toxins), Fusarium graminearum (vomitoxin or DON) Cellular level – Primary inhibition of protein synthesis Disruption of DNA and RNA synthesis Oral lesions, poor growth, abnormal feathering, decreased egg production, poor egg shell quality, regression of bursa of Fabricius Vomitoxin (DON)
    46. 46. Control (5.6 kg ) T-2 toxin 550µg/kg affected (1.9kg) Fig 7 :- Effects of T-2 on Growth of the chickens (Leeson et al., 1995) 12/11/2013 46
    47. 47. Fig 8 :- Oral Lesions on chickens caused by T-2 toxin (Leeson et al., 1995) 12/11/2013 47
    48. 48. The 40 ergot alkaloids -Claviceps purpurea can be divided into three groups: Lysergic acid (e.g. ergotamine and ergocristine) Isolysergic acid (e.g. ergotaminine) Dimethylergoline (clavines, e.g. agroclavine) Medicinally important indole alkaloids Derived from amino acid tryptophan 48 12/11/2013 (Swamy and Devegowda, 1998)
    49. 49.  Indole NH  Tryptamine NH2 CH2 CH2 NH NH2  Tryptophan CH2 NH CH COOH (Swamy and Devegowda, 1998)
    50. 50. Zearalenone • First described in US 1920s • F-2 toxin • Fusarium roseum, F. tricinctum, F. gibbosum, F. oxysporum, F. moniliforme • Source - corn, wheat, barley, oats, sorghum, tomato, beans , cucurbits, okra, spinach etc... (Huff and Ruff, 1982)
    51. 51. Phenolic compound Zearanol (zearalenol) Moisture - > 23% Temp – 12 - 25 C, Affects on reproductory system – hormonal imbalance in mammals (Huff and Ruff, 1982)
    52. 52. Effects of ZEN on humans (Sundlof and Dose 1 µg/day 6 µg/day 12.6 µg/day Strickland, 1986) Clinical Lesions Vulva swelling Smaller litter size Prevention of early featal development
    53. 53. Co- occurrence of aflatoxins, ochratoxin A and zearalenone in Capsicum powder samples available in the Spanish market L. Santos, S. Marin, V. Sanchis and A. J. Ramos Spain 2010 12/11/2013 53
    54. 54. Fig 9 : Aflatoxins, ochratoxin A and zearalenone distribution in Capsicum powder samples (Santos et al., 2010, Spain) 12/11/2013 54
    55. 55. Patulin Patulin is produced by Penicillium clariform, P. expansum, P. patulum and by Aspergillus spp Sources :- Bread, sausage, fruits (apricots, grapes, peaches, pears, and apples), and apple juice Carcinogenic toxin and responsible for subcutaneous sarcoma Affects on nervous system The allowable daily intake limit is 0.4 g/kg body weight (Smith et al., 1994) 12/11/2013 55
    56. 56. Table 6: Effect of chronic oral administration of apple juice contaminated with patulin (152.5 µg/kg) on dopamine (DA) content in the different brain areas of albino mice. (Hazmi et al., 2009, Cairo) (Hazmi et al., 2009, Cairo) 384 1.5 Pons & Medulla 508 3.5 341 2.9 630 Cerebellum 2 weeks Control Mean S.E change -11.20% 24.02% 815 4.7 1119 6.3 37.30% * ** *** significant 4 weeks Control Mean S.E change significant 6 weeks Control Mean S.E change significant Striatum 2.9 433 1.9 532 3.1 -4.95% 24.06% 824 3.1 924 503 12.14 412 3.3 660 5.0 n.s ** Cortex Hypothalamus Midbrain Hippocampus 216 1.3 655 3.7 557 402 981 7.1 296 2.5 961 4.9 575 6.3 1107 9.4 37.04% 46.72% 3.23% 12.84% *** *** n.s * 238 2.0 612 4.0 500 3.1 838 6.3 267 1.9 834 906 526 4.7 987 6.6 * 12.18% 36.27% 5.2 17.78% * *** n.s * 314 3.2 565 2.7 634 2.7 187 1.2 608 5.1 403 2.7 667 4.5 320 5.6 664 3.2 822 3.5 237 1.5 846 7.0 397 2.1 863 5.6 +1.88% 17.52 29.65 26.74% 39.14% -1.49% 29.39% n.s * *** *** *** n.s *** n.s. non-significant change according to Student,s T-test. * significant change ( P< 0.05 ) according to Student,s T-test. ** highly significant change ( p< 0.01 ) according to Student,s T-test. *** more highly significant change ( p<0.001 ) according to Student,s T-test. 12/11/2013 56
    57. 57. SCREENING OF MYCOTOXINS IN FRUITS AND VEGETABLES GROWN IN SINDH, PAKISTAN Najmus Sahar, Mubarik Ahmed, Zahida Parveen, Amber Ilyas and Abbas Bhutto Pak. J. Bot., 41(1): 337-341, 2009, Karachi 12/11/2013 57
    58. 58. Table 7: Qualitative analysis of Aflatoxin in fruits and vegetables samples by TLC method (Najmus 2009, karachi) S. No. No. of samples analyzed Observation Type of aflatoxins 1 Tomato 3 Aflatoxin B1 2 Potato 2 ND ND 3 Peas 4 ND ND 4 Beet Root 3 ND ND 5 Pumpkin 4 Aflatoxin G1 6 Garlic 3 ND ND 7 Ginger 3 ND ND 8 Onion 3 ND ND 9 Chillies (powder) 4 Aflatoxin B1 10 Carrot 2 ND ND 11 Coriander(dry) 3 Aflatoxin B1 12 Cucumber 3 Aflatoxin B1 13 Grapes 3 ND ND 14 Pomegranate 3 ND ND 15 Persimmon 3 Aflatoxin G1 16 Peanuts(dry) 4 Aflatoxin B1 17 12/11/2013 Name of sample Dates 3 ND ND 18 Peach(dry) 3 Aflatoxin B1 ND: Not Detected 58
    59. 59. Table 8:- Aflatoxin contamination in medicinally important dried rhizomes samples (Anju and Geeta, 2008, Jammu) Plants Samples analyzed Positive sample (%) AF B1 AF B2 Range of contamination (µg/g) AF B1 AF B2 Acorus calamus 26 26.9 34.6 15.55-5.11 0.14-2.01 Bergenia ciliata 24 8.3 ---- 0.10-0.61 -------- Curcuma longa 27 37 22.2 0.30-8.31 0.13-0.70 Zinziber officinale 27 11.1 14.8 0.23-0.38 0.07-0.19 AFB- Aflatoxin B 12/11/2013 59
    60. 60. Ecological aspects of distribution of potential toxin-producing micromycetes on stored apple fruit (Alma and Elena, 2006, Lithuania) • Objective :- To investigate the ecological aspects of contamination of stored apples and to elucidate the potential toxin producing micromycetes • Materials and methods • There were inspected 300 fruits (100 from each of three replicate trees) of the same maturity, size and free of physical damage and fungal infection in each control plot. The yield was kept in a storehouse with controlled storage conditions: the temperature was adjusted to 0–2 C and relative air humidity to 90% The orchard management activities and fungicide spray program were carried out according to recommendations accepted at the Lithuanian Institute of Horticulture Microbial contamination was tested immediately after apple harvesting and after three months of storage in a storehouse To isolate micromycetes, 1 cm2 pieces of apples were plated onto Malt, Czapek Dox and Potatoes Dextrose agars. Plates were incubated at 26 2 C for seven days in the dark. Grown fungi were isolated . The identification was performed according to particular manuals . The distribution frequency (%) was calculated • • • • • • • 12/11/2013 60
    61. 61. Table 9: Distribution of fungi on healthy and rotten apple fruits (Alma and Elena, 2006, Lithuania) Fungal species Distribution frequency, % Healthy fruits Acremonium roseum 4 0 Alternaria alternata 0 22 Aspergillus niger 3 17 Cladosporium cladosporioides 0 5 Cladosporium herbarum 2 10 Fusarium lateritium 0 13 Fusarium oxysporum 0 10 Penicillium biforme 4 20 Penicillium brevicompactum 10 17 Penicillium cyclopium 12/11/2013 Rotten fruits 0 28 61
    62. 62. Cont… Penicillium corylophilum 0 25 Penicillium clavigerum 0 8 Penicillium corymbiferum 0 10 Penicillium expansum 50 83 Penicillium italicum 17 67 Penicillium janthinellum 0 9 Penicillium lanosum 0 5 Penicillium roqueforti 13 28 Rhizopus oryzae 0 33 Trichoderma viride 0 38 (Alma and Elena, 2006, Lithuania) 12/11/2013 62
    63. 63. Table 10: Potentially toxin-producing micromycete species isolated from store house environment. (Alma and Elena, 2006, Lithuania) Micromycetes Air Dust Acremonium roseum - + Alternaria alternata - + Aspergillus amstelodam + - Aspergillus clavatus + - Aspergillus repens - + Aspergillus ustus + + Cladosporium cladosporioides + - Cladosporium herbarum + - Penicillium cyclopium + + Penicillium corylophilum - + Penicillium clavigerum + - Penicillium corymbiferum + - Penicillium expansum _ + Penicillium janthinellum - + Penicillium lanosum - + Penicillium oxalicum - - Penicillium roqueforti + - 10 9 12/11/2013 Total species 63
    64. 64. Mycotoxigenic fungi and mycotoxins in fast food of Bihar H. K. Chourasia, Sanjay Kumar Suman and Pramila Prasad Journal of mycology and plant pathology 38:3 2008 12/11/2013 64
    65. 65. Table 11:-Incidence of fungi (%) in raw materials of fast foods of Bihar (Chourasia et al., 2008, Bhagalpur) Raw materials of fast foods Aspergillus spp Mucor Pencillium Noodles 45.565 10.0 (5/7) 4.5 1.5 (4/7) - Cabbage 31.3 7.3 (7/10) 2.5 0.6 (3/10) Carrot 27.4 6.5 (13/20) Pizza bread Rhizopus Fusarium spp Trichoderma Others 6.5 1.2 (3/7) - - 8.5 3.5 (5/7) 15.5 4.7 (6/10) 10.4 3.0 (4/10) 4.0 0.7 (4/10) 7.5 1.5 (5/10) 4.7 1.2 (3/10) 11.5 3.5 (10/2) 10.0 2.5 (14/10) 8.9 2.5 (8/20) - 5.5 1.0 (3/20) 6.5 2.0 (6/20) 20.5 4.0 (4/8) 2.7 0.3 (3/8) 7.1 2.0 (5/8) - 10.4 2.0 4/8 Chilli sauce 15.0 3.3 (12/20) - - - 7.7 2.5 (10/20) - 14.0 4.5 (8/20) Tomato sauce 23.3 5.7 (9/15) 3.0 0.5 (6/15) - - - 6.5 1.5 (5/15) 15.5 5.7 (8/15) Maida bread 12.2 2.5 (4/7) 5.7 1.2 (3/7) - 4.5 2.1 (4/7) 12.4 2.5 (5/7) - - Burger bread 10.6 3.2 (3/7) 4.4 1.5 (4/7) - - - - 7.5 2.6 (5/7) Shimla mirch 10.0 2.7 (7/10) 4.5 1.5 (3/10) - 7.5 2.0 (6/10) - - 5.7 1.2 (2/10) Beet 5.9 1.2 (6/10) 7.8 2.5 (5/10) - 8.0 2.7 (6/10) 5.5 1.2 (4/10) 11.5 3.2 (5/10) - Egg 12/11/2013 6.5 1.5 (4/8) - 20 5.5 (5/8) 6.5 1.2 (3/8) - - 7.5 2.5 65 (4/8)
    66. 66. Fig 11 : Associaion of microbes with raw materials of fast foods a: Cabbage, b: carrot, c: Beet, d: egg, e: noodles, f:pizza bread,g: chilli sauce, h: tomato sauce, i: maida bread, j: burger bread, k: Capsicum ( Shimla mirch) 12/11/2013 66
    67. 67. 12/11/2013 67
    68. 68. 12/11/2013 68
    69. 69. Effects of Mycotoxins ZON • Irregular heats • Low conception rates • Ovarian cysts • Embryonic Loss T2-Toxin, DON, AFB1 • Gastroenteritis • Intestinal hemorrhages • Impaired rumen function • Diarrhea • Ketosis AFB1, T2-Toxin, DON • Milk contamination • Decreased milk production • Mastitis 12/11/2013 DON - Deoxynivalenol ZON – Zearalenone AFB1 – Aflatoxin B1 T2-Toxin T2-Toxin, DON • Decreased feed intake • Lower milk production • Decreased feed efficiency DON • Laminitis 69
    70. 70. Mycotoxicoses in dairy T2-Toxin: dermal lesions Zearalenone Trichothecenes: Ketosis Swollen vulva, Uterus prolapse Deoxynivalenol: Metritis affected ovaries 12/11/2013 Deoxynivalenol: inhomogeneous growth Trichothecenes: Diarrhea 70 Hypertrophy of the uterus
    71. 71. Review of genetic studies of susceptibility to facial eczema in sheep and dairy cattle C. A. Morrisa, S. H. Phuab, N. G. Cullena and N. R. Towersc AgResearch, Ruakura Research Centre, Hamilton, New Zealand; AgResearch, Invermay Agricultural Centre, Mosgiel, New Zealand Towers Consulting, Hamilton. New Zealand, 2013 12/11/2013 71
    72. 72. Fig12 :- Mean loge[GGT] of sheep groups, after dosing with the FE toxin sporidesmin and classified By the progeny-test status for FE susceptibility of their five ‘High’ (susceptible) Jersey sires or five ‘Low’ GGT (resistant) Jersey sires: results are summarized for the calves in loge i.u./l units (with bars shown for the standard error of the difference) against days since the toxin challenge (NB: Factor of 1.75 between the mean loge[GGT] values of High vs Low sire groups (74 calves)). 12/11/2013 72 (Morrisa et al., 2013, New Zealand)
    73. 73. Fig 13 :- Mean loge[GGT] of calf groups, sired by Holstein Friesian bulls that had been scored (performance tested) as weaned calves for response to FE susceptibility, or response to sporidesmin. Five such ‘High’ GGT and five ‘Low’ GGT bulls were selected for progeny testing for response to sporidesmin, alongside the progeny of eight untested/‘unselected’ Holstein Friesian bulls. Results for progeny a re shown in loge i.u./l units (with bars for standard errors of means, shown in one direction) against days since the toxin challenge) 73 (Morrisa et al., 2013, New Zealand)
    74. 74. Carry-over metabolites Poor performance Uterine infections Economic Impact Higher Incidence Diseases Cystic ovaries Immunesupression Reproductive Failure Decreased Fertility Impact Milk Displaced Abomasum, Ketosis, Retained Placenta, Metrites, Mastites, Fatty Liver, Lameness Non-response veterinary treatments ↓ Microbial growth ↓ Digestion Nutrients 12/11/2013 ↓ Rumen Motility Impaired Rumen Function 74
    75. 75. Impact of mycotoxins in milk quality 12/11/2013 75
    76. 76. Carry-over of aflatoxins in dairy Aflatoxin B1 is carried over into the milk as Aflatoxin M1 From cow… Human health concern • AfM1 is considered as “possible carcinogen” by International Agency For Research On Cancer (IARC); • Legislation limits amount of this metabolite in milk: EU: 50 ppt = 0.05 μg/kg USA: 500ppt = 0.5 μg/kg Economic concern • Milk is discarded or downgraded. 12/11/2013 …to milk! 76
    77. 77. Fig 14:- Effect of mycotoxin binder on Daily Milk production at the Jersey Farm in Bhopal. (Whitlow, 1986, Bhopal) 12/11/2013 77
    78. 78.  Neural tube birth defects  Esophageal cancer  Hepatocellular carcinoma  Carcinogenesis (Amer, 2009) 12/11/2013
    79. 79. Indigenous population that consumes high amounts of maize as their staple food Incidence of neural tube defects (NTD per 10,000 live births) General U.S. population : <3 Guatemala : 106 US Fig 15 :- Survey conducted for neural tube defects in Guatemala, South Africa and U.S.A. (Marasas et al., 2004, Guatemala) 12/11/2013 79
    80. 80. Fabio Galvano, Amedeo Pietri, Terenzio Bertuzzi, Luigi Gagliardi, Sabina Ciotti, Stefano Luisi, Matteo Bognanno, Luca La Fauci, Anna Maria Iacopino, Francesco Nigro, Giovanni Li Volti, Luca Vanella, Giuseppe Giammanco, Gabriella Lucia Tina and Diego Res. 52, Mol. Nutr. FoodGazzolo 496 – 501 2008, Italy 12/11/2013 80
    81. 81. Fig 16 : Chromatogram (HPLC; fluorescence detection) of: a) an OTA standard solution (479 ng/L), equivalent to 47.9 pg of OTA injected; b) a naturally-contaminated human milk sample containing 280 ng/L (equivalent to 56 pg of OTA injected). (Fabio et al.,2008, Italy) 12/11/2013 81
    82. 82. Fig 17 : Chromatogram (HPLC; fluorescence detection) of: a) an AFM1 standard solution (62 ng/L), equivalent to 6.2 pg of AFM1 injected; b) a naturally-contaminated human milk sample containing 72 ng/L (equivalent to 7.2 pg of AFM1 injected). 12/11/2013 (Fabio et al.,2008, Italy) 82
    83. 83. Anencephaly Spina Bifida Encephalocele Fig 18 :- Examples of suspected fumonisin-associated birth defects: anencephaly, spina bifida and encephalocele. All are from one summer in one hospital in Guatemala. Photos courtesy of Dr. Julio Cabrera (from Parrott, 2010). Parrott W (2010) Genetically modified myths and realities. New Biotechnol. 27:545-51. 12/11/2013 83
    84. 84. Spina bifida (a common form of NTD) Spina bifida (Latin: "split spine") is a developmental birth defect caused by the incomplete closure of the embryonic neural tube. Some vertebrae overlying the spinal cord are not fully formed and remain unfused and open. If the opening is large enough, this allows a portion of the spinal cord to protrude through the opening in the bones. There may or may not be a fluid-filled sac surrounding the spinal cord. Spina bifida 12/11/2013 84 (Source:http://en.wikipedia.org/wiki/Spina_bifida)
    85. 85. Encephalocele: which results when other parts of the brain remain unfused. Anencephaly : a condition in which the portion of the neural tube which will become the cerebrum does not close, 12/11/2013 85 (Source: http://neuropathology.neoucom.edu/chapter11/images11/11-2al.jpg)
    86. 86. LUNG TUMORS Adenocarcinoma Large Cell Aflatoxin Positive Mesothelioma Ochratoxin found in chest wall, diaphragm. No ochratoxin found in pleura 12/11/2013 Bronchial Alveolar CA (BAC) Aflatoxin Positive Ochratoxin Positive 86
    87. 87. Removal or Elimination of Mycotoxins • Pre harvest control • Resistant varieties • Crop rotation Inactivation of Mycotoxins • Adequate irrigation Removal of Mycotoxins During • Control of insect pests. Food Processing. • Postharvest Control • 10–13% moisture Avoiding Human Exposure • Avoid insect and rodent infestation • Methods of harvesting and storage Detection and Screening of Mycotoxins Dietary Modifications 12/11/2013 87
    88. 88. Conclusion............!!!!! • Mycotoxins are highly stable compounds and ubiquitous • Causes severe crop loss because contamination can occur both before and after harvest • Severe health hazards to Animals and humans • Good farm management practices and storage • Regulatory commissions should develop • The development of sophisticated kits for the detection of minute amounts of mycotoxins is the most important • Create awareness and education 12/11/2013 88

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