Graduation Project ( Presentation Slides ) About Incidence of AFLATOXIN M1 in MILK

1. Incidence Of
AFLATOXIN M1
in Milk
A Graduation Project Presented By:
Mohammed Azzaz Ghamri Ibraheem
Food Quality & Control Program (FQCP)
Faculty of Veterinary Medicine Benha University
Under Supervision of
Prof. Fahim Shaltout Prof. Adham M. Abdou
Professor of Meat Hygiene Professor of Milk Hygiene
Food Control Department, Fac.Vet.Med, Benha University

2. Prof. Mohamed Abu Salem
HassaneinProf. Faten

3. General Background
Biological Data
Analytical Methods
Sampling Protocols
Food Consumption and Dietary Intake Estimates
Levels and Patterns of Contamination of Food Commoidites
Effects of Processing
Adverse Effect
Dose–response Relationship and Estimation of
Carcinogenic Risk

5. AFLATOXINS from Aspergillus spp
(A. flavus, A.parasiticus and the rare A.Nomius )
Aflatoxin B1 (AFB1)
Most Potent biological effects**
Aflatoxin M1 (AFM1) and Aflatoxin M2 (AFM2)
are the hepatic hydroxylated metabolites
of AFB1 and AFB2
AFB1 per se is not genotoxic but is but is
bioactivated by liver metabolism to increases its
hydrophilic properties and excretion in urine or
milk
AFM1is found in milk and milk
products obtained from livestock that have
ingested AFB1-contaminated feed
Introdution

6. The Aim
of The
Subject
What Is
AFLATOXIN
M1
Incidence
&
Occurance
Direct &
Indirect
reasons
Pathoo-
genicity
Toxicity
&
Adverse
Effect
Community
awareness
Prevention
Control
Measurement &
Treatment
Stability &
Resistance

7. ** General Background
Aflatoxins can be produced by three species of Aspergillus—A.
flavus, A. parasiticus, and the rare A. nomius—which contaminate
plants and plant products.
A. flavus produces only B aflatoxins, while the other two species
produce both B and G aflatoxins. Aflatoxins M1 and M2 are the
hydroxylated metabolites of aflatoxins B1 and B2 and can be
found in milk or milk products obtained from livestock that have
ingested contaminated feed. The main sources of aflatoxins in
feeds are peanut meal, maize and cottonseed meal.

8. **BIOLOGICAL DATA
1 – Chemical Structure

9. **BIOLOGICAL DATA
2 - Biochemical aspects:A- Biotransformation:
B- Effects of oltipraz and ethoxyquin

10. **ANALYTICAL METHODS
1- Screening tests :
Early detection of aflatoxin M1 and
removal of small lots of
contaminated milk can prevent
contamination of much larger
volumes. Screening methods are
particularly useful if they can be
carried out quickly, easily, and
economically. They should allow
detection of concentrations of
aflatoxin M1 in milk as low as those
detected by the ultimate
quantitative methods

11. **ANALYTICAL METHODS
:2- Quantitative methods
The quantitative methods that have been
developed and validated for aflatoxin M1 in
milk and milk products were originally
designed to analyse milk powder. Milk was
spray-dried or lyophilized to preserve its shelf
life and to reduce sample bulk.
*A more recent advance in quantitative
extraction of aflatoxin M1 and subsequent
clean-up is use of immunoaffinity cartridges.
These columns are composed of monoclonal
antibodies specific for aflatoxin M1, which are
immobilized on Sepharose® and packed into
small cartridges (see Figure 2). The first
published method for aflatoxin M1 with
immunoaffinity columns was that of Mortimer
et al. (1987).

12. **ANALYTICAL METHODS
3- Analytical quality
assurance
The availability of collaboratively studied
‘official’ methods of analysis for aflatoxin M1,
with acceptable performance characteristics,
is no guarantee of accurate results. Check
sample programmes for aflatoxins, including
aflatoxin M1 in milk, organized by IARC
(Friesen & Garren, 1982) have shown that
there can be wide variation in results. In
compliance with the principles of analytical
quality assurance, measurements of the
mycotoxin by different laboratories should be
reliable and comparable. A quality assurance
programme includes, when possible, use of
(certified) reference materials and many other
elements.

13. **SAMPLING PROTOCOLS
Two effective methods for
controlling aflatoxin M1 in the food supply are
to sample dairy feed for aflatoxin B1 or to
sample the milk directly for aflatoxin M1.
but there has been little evaluation of
sampling plans to detect aflatoxin M1 in milk.
It might be difficult to design an effective
programme to control aflatoxin in granular
feed, particularly at low concentrations,
because of its heterogeneous distribution in
these commodities, which results in wide
sampling variation.
In the absence of information on the efficacy
of sampling plans for the determination of
aflatoxin M1 in milk, it is recommended that
the European model, in which a 500-g sample
composed of five 100-g portions of milk is
taken from a batch, be used for the minimum
sample size and sample selection method.

14. **FOOD CONSUMPTION AND
DIETARY INTAKE ESTIMATES
Dietary intake of aflatoxin
B1 was estimated from
data on the concentrations
of aflatoxin M1 in milk
submitted to FAO/WHO,
from selected reports in
the literature, and from
data on milk consumption
in the GEMS/Food
Regional Diets (WHO,
1998).

15. **FOOD CONSUMPTION AND
DIETARY INTAKE ESTIMATES
* Effect of proposed maximum levels in
:foods on dietary intake of aflatoxin M1

16. ** Toxicity
Aflatoxin M
1
is cytotoxic, as demonstrated
in human hepatocytes in vitro and its
acute toxicity in several species is similar
to that of aflatoxin B
1
. In ducklings and
rats, the acute and short-term toxicity of
aflatoxin M
1
was similar to or slightly less
than that of aflatoxin B
1
. In studies of
carcinogenicity, aflatoxin M
1
was about
one order of magnitude less potent than
aflatoxin B
1
, even in sensitive species like
the rainbow trout and the Fischer rat. The
in vitro genotoxic potency of aflatoxin
M
1
was similar to that of aflatoxin B
1
in
some test systems and between one-half
and one-sixth that of aflatoxin B
1
in other
test systems.

17. **EFFECTS OF PROCESSING

18. **EFFECTS OF PROCESSING
Fate of aflatoxin M1 during
processing of milk:
Treatments that are common in
the dairy industry can be
separated into two distinct
processes: those that do not
involve separation of milk
components, such as heat
treatment, low-temperature
storage, and yoghurt preparation;
and processes that involve
separation of milk components,
such as concentration, drying,
and cheese and butter production.

19. **EFFECTS OF PROCESSING
The stability of aflatoxin M1 during
heat processing, such as
pasteurization and heating milk
directly on a fire for 3–4 h has been
studied
Studies of the stability of aflatoxin
M1 in milk during cool or frozen
storage gave variable results ,but
storage of frozen contaminated
milk and other dairy products for a
few months did not appear to affect
the aflatoxin M1 content.
The manufacture of cultured dairy
products, such as kefir and
yoghurt, also did not significantly
decrease the aflatoxin M1 content

20. **EFFECTS OF PROCESSING
Degradation of aflatoxin
M1 in milk:
The observation that the processes
described above do not generally lead
to loss of aflatoxin M1 is of
considerable practical importance.
Several possibilities for eliminating or
inactivating aflatoxin M1 in milk,
involving chemical and physical
treatment, have been investigated. The
chemicals that have been studied for
their ability to degrade aflatoxin M1 are
limited to those that are permitted as
food additives: sulfites, bisulfites, and
hydrogen peroxide

21. **EFFECTS OF PROCESSING
The chemical and physical
treatments described are not
readily applicable in the dairy
industry, at least at present, as
little is known about the biological
safety, or the nutritional value of
the treated products. Moreover,
the costs of the processes may be
considerable and prohibitive for
large-scale application. If aflatoxin
M1 cannot be destroyed or
removed readily, it can be
excluded from milk only by
eliminating aflatoxin B1 from the
diet of animals.

22. ** Adverse Effect

23. ** Adverse Effect
aflatoxin can appear in the meat, milk, and eggs of food animals
that have consumed aflatoxin-contaminated feeds. The aflatoxin
group consist of B1, B2, G1, and G2. Aflatoxin B1 is the most
potent of the group and has been shown to be a potent
carcinogen. Aflatoxin M1 is metabolic breakdown product of
aflatoxin B1 and can appear in the milk of lactating cows
consuming significant quantities of aflatoxin B1. Aflatoxin M1 is
not as carcinogenic as B1 but can be as toxic. The conversion
of aflatoxin B1 in feed to aflatoxin M1 in milk is about 1% to 2%.
The FDA regulatory level of aflatoxin in milk is 0.5 ppb.

24. ** Adverse Effect
High-level aflatoxin exposure produces an acute hepatic necrosis,
resulting later in cirrhosis, or carcinoma of the liver. Acute hepatic
failure is made manifest by hemorrhage, edema, alteration in digestion,
changes to the absorption and/or metabolism of nutrients, and mental
changes and/or coma.
No animal species is immune to the acute toxic effects of aflatoxins;
however, adult humans have a high tolerance for aflatoxin exposure
and rarely succumb to acute aflatoxicosis.
Chronic, subclinical exposure does not lead to symptoms as dramatic
as acute aflatoxicosis. Children, however, are particularly affected by
aflatoxin exposure, which leads to stunted growth and delayed
development. Chronic exposure also leads to a high risk of developing
liver cancer.
Medical research indicates that a regular diet including apiaceous
vegetables such as carrots, parsnips, celery and parsley, may reduce
the carcinogenic effects of aflatoxin.
Moreover, aflatoxin B1 can permeate through the skin. Dermal
exposure to this aflatoxin in particular environmental conditions can
lead to serious health risks.

25. ** Adverse Effect

26. ** Prevention & Control
there are several possibilities for
preventing the presence of aflatoxin
M1 in dairy products; each has its
advantages and disadvantages.
Although prevention of contamination
of dairy cattle feed is the ideal, it may
not be possible in practice. Various
decontamination methods have a role
to play in preventing and reducing the
concentrations of aflatoxin M1 in dairy
products.

27. The most effective way of
controlling aflatoxin M1 in the food
supply is to reduce contamination
with aflatoxin B1 of raw materials
and supplementary feedstuffs for
dairy cattle. Specific regulations
exist in many countries (FAO,
1997), and practical programmes
are being developed; e.g. the
Codex Committee on Food
Additives and Contaminants has
developed a code of practice for
reducing aflatoxin B1 in raw
materials (van Egmond et al.,
** Prevention & Control

28. ** Prevention & Control
Reduction can be achieved by good manufac-turing practices and
good storage practices. If preventive measures fail to reduce fungal
growth and aflatoxin B1 formation in agricultural commodities
intended for use as animal feeds, the last means for avoiding or
reducing the occurrence of aflatoxins in feed is to eliminate (part of)
the toxins. Feeds that have higher concentrations of aflatoxin B1 may
be acceptable for feeding to dairy animals if they are blended with
feed that has lower concentrations, provided that the resultant
aflatoxin M1 concentration in milk does not exceed levels considered
to be safe. In principle, aflatoxin-contaminated consignments of feeds
can be decontaminated by removing the toxin (segregation) or by
converting it to a non-toxic form (degradation).

29. ** Prevention & Control
Degradation may be achieved by physical, chemical, or biological means.
Attempts have been made to degrade aflatoxins in feed by applying physical
treatments such as heat, microwaves, gamma-rays, X-rays, ultra-violet light
and adsorption (van Egmond & Speijers, 1999). Degradation of aflatoxin
M1 has also been attempted by combined treatments, such as ultra-violet
radiation followed by ultrafiltration. In most cases, neither heat treatment not
irradiation is effective. Adsorption of aflatoxins from animal feed onto
bentonite and hydrated sodium calcium aluminosilicate (Veldman, 1992;
Galvano et al., 1996a) has been used in the feed industry to reduce the
aflatoxin M1 content of milk (Harvey et al., 1991; Phillips et al., 1995).

30. ** Prevention & Control
A newer approach is use of oltipraz,
a substituted dithiolthione that
inhibits aflatoxin B1 metabolism by
inhibiting the activity of several
cytochrome P450 enzymes
(Kuilman et al., 2000). No aflatoxin
M1 formation was found in bovine
hepatocytes incubated with
aflatoxin B1 and oltipraz. The
findings suggest that oltipraz is
highly effective in inhibiting
aflatoxin M1 contamination of milk
from diary cows exposed to
aflatoxin B1-contaminated feeds.

31. ** Prevention & Control
Some chemical procedures have been
developed to degrade aflatoxins in
animal feed, usually based on addition
of oxidizing agents, aldehydes, acids,
and bases. The most widely used
chemical detoxication reagent is
ammonia, as an anhydrous vapour or
as an aqueous solution. Treatment of
aflatoxin B1 with ammonia opens the
lactone ring of the molecule.
Ammoniation of agricultural
commodities leads to decomposition
of 95–98% of the aflatoxin B1 present.
This process is used in various
countries for the decomposition of
animal feedstuffs

35. ** Conclusion
Aflatoxin M1 have a lot of bad effects
which more toxic for human body and
may cause liver of lung carcinoma
,degeneration of DNA and stunted
growth , hepatic diseases,,,. .

36. ** Conclusion
The most important part of treatment of
AFLATOXIN M1 is have a good hygienic
practice in handling and treating of the
animal feed (transfer, storage,….) to prevent
of AFLATOXIN B1 (Most Carcinogenic) which
transformation to AFLATOXIN
M1(Toxic)which secreted in milk ,urine.
There are different ways to prevent or
treatment of reduce of AflaToxin M1 which
may physical ,chemical ,biological or
.mechanical processes

37. ** Recommendation

38. ** Recommendation

39. Regards ,,,