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FABAD J. Pharm. Sci., 33, 51–66, 2008
51
REVIEW ARTICLE
A Special Focus on Mycotoxin Contamination in
Baby Foods: Their Presence and Regulations
Pınar ERKEKOĞLU*, Gönül ŞAHIN*, Terken BAYDAR*°
A Special Focus on Mycotoxin Contamination in Baby
Foods: Their Presence and Regulations
Summary
Food safety is one of the major concerns in researches related
to food toxicology. Contaminants present in food and feed
are the most attention-drawing subjects in the last decade.
Particularly, mycotoxin contamination is of great importance
as it is widespread and unpreventable. Mycotoxins are toxic
secondary metabolites by different fungi species. These
compounds pose a potential threat to human and animal
health through the ingestion of food products prepared
from these commodities. Mycotoxicosis is the term used for
poisoning associated with exposures to mycotoxins. The
symptoms of a mycotoxicosis depend on the type of mycotoxin;
the concentration and length of exposure; as well as age,
health, and sex of the exposed individual. Aflatoxin B1 and
ochratoxin A are mutagenic, teratogenic, and carcinogenic
in many species where Fusarium toxins such as T2 toxin
pose a threat as biological warfare agent. Many international
agencies are trying to achieve universal standardization of
regulatory limits for mycotoxins. Special emphasis must be
drawn to mycotoxin contamination of baby foods and infant
formulas as babies and small children are the most susceptible
population to the effects of these toxins. In this review, the
toxic effects of mycotoxins, the regulations in Europe and
United States as well as Turkey and particularly the studies
and regulations in baby foods will be dwelt upon.
Key Words: mycotoxin, regulatory limits, baby food/infant
formula
Received: 26.11.2009
Revised: 25.05.2010
Accepted: 03.07.2010
Bebek Mamalarında Mikotoksin Kontaminasyonuna
Bakış: Bulunuşları ve Yasal Düzenlemeleri
Özet
Gıda güvenliği gıda toksikolojisi ile ilgili araştırmaların
ana konularından biridir. Gıdalarda ve yemlerdeki
bulaşıcılar son on yılın en çok ilgi çeken konusu olmuştur.
Yaygın ve önlenemez olması nedeniyle özellikle mikotoksin
kontaminasyonu çok önemlidir. Mikotoksinler farklı mantar
türlerinin toksik sekonder metabolitleridir. Bu bileşikler
bunları içeren gıdaların alımıyla insan ve hayvan sağlığı
için potansiyel bir tehdittir. Mikotoksikoz mikotoksin
maruziyetiyle ortaya çkan zehirlenme için kullanılan bir
terimdir. Mikotoksikozisin semptomları mikotoksin türüne,
maruziyetin miktarına ve süresine, bireyin yaşına, sağlık
durumuna ve cinsiyetine göre değişmektedir. Aflatoksin B1
ve okratoksin A mutajenik, teratojenik ve karsinojeniktir.
Fusarium türlerinden T2 biyolojik savaş ajanı olarak
bir tehdit oluşturmaktadır. Birçok uluslararası kuruluş
mikotoksinlerin limit düzenlemelerine bir standardizasyon
getirmeye çalışmaktadır. Bebekler ve küçük çocuklar bu
toksinlerin etkilerine daha hassas oldukları için özel olarak
bebek mamalarındaki mikotoksin kontaminasyonuna
dikkat edilmelidir. Bu derlemede mikotoksinlerin toksik
etkilerinden; Avrupa, Amerika Bileşik Devletleri ve
Türkiye’deki regülasyonlardan bahsedilecek ve özellikle bebek
mamaları üzerindeki çalışmalar ve regülasyonların üzerinde
durulacaktır.
Anahtar Kelimeler: mikotoksin, düzenleyici limitler, bebek
maması
* Department of Toxicology, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey
° Corresponding author E-mail: tbaydar@hacettepe.edu.tr
Erkekoğlu, Şahin, Baydar
52
INTRODUCTION
A major international focus has been ensuring the
safety of food. Toxins present in food and animal feed
are of major concern of health care givers and public
for decades. A toxin can be defined as a substance
that is synthesized by a plant species, an animal, or by
microorganisms, that is harmful to another organism.
The term ‘mycotoxin’ is usually reserved for the
relatively small (MW 700), toxic chemical products
formed as secondary metabolites by a few fungi that
readily colonize crops in the field or after harvest.
These compounds pose a potential threat to human
and animal health through the ingestion of food
products prepared from these commodities. Generally,
crops that are stored for more than a few days become
a potential target for mould growth and mycotoxin
formation. Mycotoxins can occur both in temperate
and tropical regions of the world, depending on the
species of fungi. Contamination can occur pre- or post-
harvest or at the field (1). Favorable conditions such as
high humidity and high temperature can increase the
content of mycotoxin during storage. Cereals, spices,
nuts, grapes, apples, dried fruit, dried vegetables
(peas, beans), oil seeds, teas, cocoa and coffees can
contain high amount of different mycotoxins. Food-
based mycotoxins and their health effects were
extensively studied in the last century and there are
several regulations based on their presence in different
foodstuffs (2-4). Mycotoxins can also enter the human
food chain via meat or other animal products such as
eggs, milk and cheese as the result of livestock eating
contaminated feed (5).
Mycotoxin Types
Aflatoxins
Aflatoxins (AFs) are naturally occurring highly
toxic mycotoxins that are produced as secondary
metabolites of different widespread Aspergillus
species (Aspergillus flavus, Aspergillus parasitius,
Aspergillus nomius) and they may be present in
groundnuts, other edible nuts, dried fruits, spices,
figs and cereals (especially maize) (2, 6-10). Sources
of AF contamination in animal feedstuffs may vary
geographically. Contamination of agricultural
crops with AFs is a worldwide problem not limited
to developing countries, where both climatic
and technological conditions stimulate aflatoxin
formation (11). Aspergillus flavus produces AFB1 and
AFB2, while two other species produce AFG1 and
AFG2 (10). The diseases caused by AF consumption
are called aflatoxicosis. Acute aflatoxicosis results
in death; chronic aflatoxicosis results in cancer,
immune suppression, and other “slow” pathological
conditions (12).
AFB1 is the most known potent natural carcinogen.
The activation of AFB1 by Phase I enzymes namely,
cytochrome P450 (CYPs) isoenzymes CYP1A2 and
CYP3A4, produces AF1-8,9-epoxide which is highly
carcinogenic in humans. It forms DNA adducts
and albumin adducts (13). A reactive glutathione
S-transferase (GST) system found in the cytosol and
microsomes catalyzes the conjugation of activated
aflatoxins with reduced glutathione, leading to the
excretion of aflatoxin (14). Variations in the level
of the GST system as well as variations in the CYP
system are thought to contribute to the differences
observed in interspecies aflatoxin susceptibility (15).
AFB1 is listed as Group I agent by International
Agency for Research on Cancer (IARC). The principle
target organ of AFB1 is liver. It is known that the
reactive aflatoxin epoxide binds to the N
7
position
of guanines (16). AFB1 causes mutation of p53 gene
at third base of codon 249, and takes the form of G
>T transversions. This mutation may inactivate p53
and the detection of TP53 mutant DNA plasma is a
biomarker of both AFB1 exposure and hepatocellular
carcinoma (17, 18). AFB1 alters the activation of p53
in CYP450-expressing human lung cells (19). Long-
term exposure to AFB1 produces liver enlargement
and hepatocellular carcinoma (HCC), which is one
of the most common cancers worldwide, causing
millions of deaths annually (2, 13, 17, 20). There are
several studies conducted on animals indicating the
carcinogenic potency of AFB1. Mice and hamster
are protected against AFB1-induced HCCs in in vivo
and in vitro conditions. Mouse liver is protected as a
consequence of the impermeability of mitochondrial
membrane to the toxin. On the other hand, it is a more
complex process including both a permeability barrier
and a possible scavenging system in hamsters (21).
Moreover, AFB1 causes colon and kidney cancers in
rats, lung adenomas in mice, cholangiocellular cancer
FABAD J. Pharm. Sci., 33, 51–66, 2008
53
in hamsters, osteogenic sarcoma, adenocarcinoma of
gall bladder and pancreas cancer in monkeys (22).
Antibodies to AFB1 have been reported in humans
and they are considered to be an indicator of
exposure (23). Besides, AFs cause increased levels of
tumor necrosis factor alpha (TNF-a) and changes in
serum lactate dehydrogenase activity (24). AFs are
also suspected to cause Reye-like syndrome with
multiple symptoms. Moreover, AFs target kidney
and cause renal cortex changes (25).
AF exposure cause changes in oxidative phosphor-
ylation, which subsequently cause changes in the
structure of mitochondria (abnormal mitochondrial
structure, and elevation in mitochondrial enzymes)
(25-30). The changes in mitochondria are important in
aflatoxin-induced hepatocarcinogenesis as AFs pref-
erably attack mitochondrial DNA (mtDNA) three to
four times higher than nuclear DNA (30). Aflatoxins
also cause mitochondria-directed apoptosis (31).
The “no observed adverse effect level (NOAEL)”
for AFB1 in male CD-1 mice, male BALB/c mice
and male C57B1/6 mice is found to be 30 µg/kg
b.w. and in male weanling rats it is 60 µg/kg b.w.
(32-34). In studies performed on Gambian children,
considering the impairment in host resistance to
infections, the NOAEL value was found to be 30 µg/
kg b.w. (35, 36). However, the European Food Safety
Authority (EFSA) panel on mycotoxins could not
establish a NOAEL as a point of departure for the
risk assessment (37).
AFM1 is the metabolite of AFB1 in milk of cattle fed
on contaminated foods. AFM1 may be present in
animal organs and tissues, e.g. kidneys, and in animal
products, e.g. milk, milk powder, cheese, butter and
other dairy products after consumption of AFB1-
contaminated feeds by animals (11). A tolerable daily
intake (TDI) of 0.2 ng/kg b.w. for AFM1 was calculated
by Kuiper-Goodman (38) and it has been categorized
by the IARC as a Group IIB, a possible human
carcinogen. In the assessment of carcinogenicity, the
infants are more exposed to the risk because the milk
is a major constituent of their diet. It must also be
considered that young animals have been found to
be more susceptible to AFB1 (and so probably AFM1)
than adults. Therefore, the presence of AFM1 in milk
and milk products is considered to be undesirable
(39-43). AFM1 is cytotoxic, as shown in human
hepatocytes in vitro and its acute toxicity in several
species is similar to that of AFB1. AFM1 can also
cause DNA damage, gene mutation, chromosomal
anomalies and cell transformation in mammalians
cells in vitro, in insects, lower eukaryotes and bacteria.
However, AFM1 is less mutagenic, and genotoxic
than AFB1 (44-47).
Ochratoxins
Ochratoxins A, B, and C are mycotoxins produced by
some Aspergillus species and Penicillium species, like
Aspergillus ochraceus or Penicilium viridicalum, with
OTA as the most prevalent and relevant fungal toxin
of this group. The mostly debated toxin of this group
of mycotoxins is OTA (48). OTA was discovered as
a metabolite of Aspergillus ochraceus in 1965 (48).
OTA is known to occur in commodities like cereals,
coffee, dried fruit and red wine. Besides, OTA is of
special interest as it can be accumulated in the meat
of animals. OTA-contaminated feed has its major
economic impact on the poultry industry. Chickens,
turkeys and ducklings are susceptible to this toxin.
OTA has been detected in blood and other animal
tissues and in milk, including human milk (49).
The biological effects of OTA are well documented.
IARC has classified OTA as a possible carcinogen
(Group 2B) (50). There have been reports on its
immuno-suppressive nature (51), teratogenicity
(52), reproductive toxicity (55), mutagenicity and
carcinogenicity (52, 56). OTA is a nephrotoxin to all
animal species studied to date and is most likely
toxic to humans, who have the longest half-life for its
elimination of any of the species examined (57). OTA
disturbs cellular physiology in multiple ways, but it
seems that the primary effects are associated with
the enzymes involved in phenylalanine metabolism,
mostly by inhibiting the enzyme involved in the
synthesis of the phenylalanine-tRNA complex
(58, 59). In addition, it inhibits mitochondrial ATP
production (58) and stimulates lipid peroxidation (61).
It has also been hypothesized that the heterozygous
gene pattern for phenylketonuria might occur in
relatively high frequency and it is an advantage in
Erkekoğlu, Şahin, Baydar
54
the ochratoxin poisoning (62) and that OTA might be
a risk factor for testicular cancer (63).
Exposure to OTAs through diet can cause acute
nephrotoxicity, and may be carcinogenic (55). Several
studies in literature have suggested a correlation
between exposure to OTA and Balkan endemic
nephropathy (BEN), a chronic tubulointerstitial
disease, found between 0.5 to 4.4 % (in some places as
high as 20%) in South-Eastern Europe (Serbia, Bosnia
and Herzegovina, Croatia, Romania, and Bulgaria).
More specifically, BEN is most likely to occur among
those living along the confluence of the Danube River,
a region in which the plains and low hills generally
have high humidity and rainfall. These conditions
seem to contribute to high occurrence of OTA in
food and feed. A high frequency of urothelial atypia,
occasionally culminating in tumors of the renal pelvis
and urethra, is associated with this disorder (56).
The most comprehensive studies on OTA toxicity in
rats have been performed within the US National
Toxicology Program (US-NTP) (63). The overall
NOAEL level derived from these studies was found
to be was 21 μg/kg b.w. per day for 5 days/week,
equivalent to 15 μg/kg b.w. per day (64, 65). On the
other hand, pigs were found to be more susceptible
to OTA toxicity and the NOAEL level in female pigs
was reported as 8 μg/kg b.w. per day. In consideration
of these findings, the EFSA Panel in year 2006 tried to
find the lowest observed adverse effect level (LOAEL)
and NOAEL for OTA in humans by applying a set of
uncertainty factors; however, these levels have not
yet been set for humans (66).
Patulin
Patulin is a mycotoxin produced by a variety of
molds, particularly Aspergillus and Penicillium. It is
commonly found in rotting apples, and the amount
of patulin in apple products is generally viewed
as a measure of the quality of the apples used in
production. It is not a particularly potent toxin, but
a number of studies have shown that it is genotoxic,
which has led to some theories stating that it may be
a carcinogen, though animal studies have remained
inconclusive. IARC has classified patulin as a Group
III carcinogen (a compound for which there is not
enough data to allow its classification) (67). Several
countries have instituted patulin restrictions in
apple products. The World Health Organization
(WHO) recommends a maximum concentration of 50
µg/L in apple juice (68).
The maximum provisional
tolerable daily intake (PTDI) of 0.43 μg/kg b.w./
day. NOAEL value in rats was found to be 0.3 mg/
kg b.w./day in rats (69). Several studies have been
conducted on patulintent of apple juices marketed in
Turkey. In a study performed by Gökmen and Acar,
the researchers found that throughout the years the
concentrations of patulin in apple juices decreased.
Percentages of concentrates exceeding the maximum
permitted concentration of 50 µg/L were 52%, 34%,
8% and 8% for 1996, 1997, 1998 and 1999, respectively
(70). In another study performed by Yurdun et al., 40%
of the apple juice samples had patulin contamination
levels higher than 50 µg/L (71).
Fusarium Toxins
A variety of Fusarium fungi, which are common soil
fungi, produce a number of different mycotoxins of
the class of trichothecenes: T-2 toxin, HT-2 toxin, de-
oxynivalenol and nivalenol and some other toxins
zearalenone and fumonisins. The Fusarium fungi are
probably the most prevalent toxin-producing fungi
of the northern temperate regions and are commonly
found on cereals grown in the temperate regions of
America, Europe and Asia. Fusarium toxins have been
shown to cause a variety of toxic effects in both ex-
perimental animals and livestock. In some occasions,
toxins produced by Fusarium species have also been
suspected to have caused toxicity in humans (72).
a. Fumonisins
Fusarium verticillioides and the related Fusarium
proliferatum are the only fungi that produce
significant quantities of fumonisins. Fumonisins
occur in sorghum, asparagus, rice, beer and mung
beans infrequently (72). Conditions favoring
fumonisin production appear to include a period of
drought during the growing season with subsequent
cool, moist conditions during pollination and kernel
formation. Equine leukoencephalomalacia, caused
by high exposure to fumonisins, is a disease of the
central nervous system (CNS) that affects horses,
mules, and donkeys (73).
FABAD J. Pharm. Sci., 33, 51–66, 2008
55
Fumonisins are classified as Fumonisin B1,
Fumonisin B2 and Fumonisin B3 where Fumonisin
B1 is the most prevalent member and Fumonisin
B3 is relatively There is inadequate evidence in
humans for the carcinogenicity of fumonisins. There
is sufficient evidence in experimental animals for
the carcinogenicity of fumonisin B1. It is classified
as “possibly carcinogenic to humans (Group
IIB)” by IARC (74). Fumonosin B1 is hepatotoxic
and nephrotoxic in all animal species tested (68).
Fumonisin were not found to be genotoxic (72).
Concerning chronic exposure when liver toxicity is
taken as the end point, the NOAEL level is given as
0.6 mg/kg b.w./day for rats. When kidney toxicity is
the end point in rats, NOAEL level was given as 0.25
mg/kg b.w./day by US-NTP (75).
b. Trichothecenes
The trichothecenes constitute a family of more than sixty
sesquiterpenoid metabolites produced by a number
of fungal genera, including Fusarium, Myrothecium,
Phomopsis, Stachybotrys, Trichoderma, Trichothecium,
and others (76, 77). The term trichothecene is derived
from trichothecin, which was the one of the first
members of the family identified. They are commonly
found as food and feed contaminants (78-80). The
symptoms produced by various trichothecenes
include effects on almost every major system of the
vertebrate body; many of these effects are due to
secondary processes that are initiated by often poorly
understood metabolic mechanisms related to the
inhibition of protein synthesis (81). Of the naturally
occurring trichothecenes, T-2 and diacetoxyscirpenol
appear to be the most potent in animal studies. In
addition to their cytotoxic activity, they have an
immunosuppressive effect that results in decreased
resistance to infectious microbes (82, 83). They cause
a wide range of gastrointestinal, dermatological, and
neurologic symptoms (84). (DON and T-2 are the best
studied of the trichothecenes produced by Fusarium
species.
c. Deoxynivalenol
Deoxynivalenol (DON) is a common mycotoxin found
in grains. DON may have adverse health effects after
acute, short-term, or long-term administration. After
acute administration, deoxynivalenol produces two
characteristic toxicological effects: decrease in feed
consumption (anorexia) and emesis (vomiting) (78).
When ingested in high doses by agricultural animals, it
causes nausea, vomiting, and diarrhea; at lower doses,
pigs and other farm animals exhibit weight loss and
food refusal (83). For this reason, DON is sometimes
called “vomitoxin” or “food refusal factor”. Although
less toxic than many other major trichothecenes, it is
the most prevalent and is commonly found in barley,
corn, rye, safflower seeds, wheat, and mixed feeds
(85). In 1993, IARC placed deoxynivalenol in Group
III, not classifiable as to its carcinogenicity to humans
(86). The no-observed-effect level (NOEL) is 0.1 mg/
kg b.w./per day (85). On the other hand, NOAEL for
DON is found as100 μg/kg b.w./ per day (87).
d. T-2 Toxin
Among the naturally-occurring trichothecenes
found in food and feed, T-2 toxin is the most potent
and toxic mycotoxin. Corn, wheat, barley, oats, rice,
rye and other crops have been reported to contain
T-2 toxin (88). Toxin production is greatest with
moisture and temperatures and adequate storage
with low moisture and insect control will minimize
further fungal growth and T-2 toxin production
(89). The major effect of T-2 toxin and other
trichothecenes is that they inhibit protein synthesis
which is followed by a secondary disruption of DNA
and RNA synthesis. It affects the actively dividing
cells such as those lining the gastrointestinal tract,
skin, lymphoid and erythroid cells. It can decrease
antibody levels, immunoglobulins and certain other
humoral factors such as cytokines (90, 91). The
manifestations of disease include signs of weight
loss or poor weight gains, bloody diarrhea, dermal
necrosis or beak and mouth lesions, hemorrhage
and decreased production of milk and eggs.
Besides, it characteristically causes aleukia which is
an absence or extreme reduction in the number of
white blood cells in circulating blood (92, 93). No
effect (NOAEL) was observed in rats was found to
be 0.5 mg/kg b.w./day by Sirkka et al (1992) et al.
(94). On the other hand, for mice the NOAEL value
was found to be 0.23 mg/kg b.w/per day (1994).
The EFSA panel in 2001 after applying a set of
uncertainty factors set the NOAEL forT-2 toxin as
0.06 µg/kg b.w./day (95).
Erkekoğlu, Şahin, Baydar
56
e. Zearalenone
Zearalenone is also named as RAL, FES, Compound
F-2 or Toxin F2 and is a potent estrogenic metabo-
lite produced by some Giberella species. Zearalenone
is the primary toxin causing infertility, abortion or
other breeding problems, especially in swine (96-99).
Studies of the pharmacokinetics and metabolism of
zearalenone demonstrate that it is extensively me-
tabolized by intestinal tissue in pigs, and possibly in
humans, during its absorption, with the formation
of a- and β-zearalanol, which are subsequently con-
jugated with glucuronic acid. Zearalenone has been
tested for genotoxicity in a variety of test systems
and the results were negative, except for the induc-
tion of chromosomal aberrations after exposure of
mammalian cells in vitro to very high concentrations.
Hepatocellular adenomas and pituitary tumors were
observed in studies of long-term toxicity and carcino-
genicity in mice, but only at doses greatly in excess of
the concentrations that have hormonal effects, i.e. at
8-9 mg/kg b.w./per day or more. These tumors were
a consequence of the estrogenic effects of zearalenone
(96-99). The NOAEL value of zearalenone was given
as 4 mg/kg/per day in rats (100).
Regulations for Mycotoxins in Food
Risk Assessment for Mycotoxins and Worldwide
Mycotoxin Regulations
The number of countries regulating mycotoxins has
increased significantly over the years. Regulations
have become more diverse and detailed with newer
requirements with regard to official procedures for
sampling and analytical methodology (101). At least
99 countries had mycotoxin regulations for food
and/or feed in 2003, an increase of approximately
30% compared with 1995. All countries with
mycotoxin regulations in 2003 had regulatory limits
for at least AFB1 or the sum of AFB1, B2, G1, and
G2 in food and/or feed. Specific regulations also
exist for several other mycotoxins such as AFM1; the
trichothecenes DON, diacetoxyscirpenol, T-2 toxin
and HT-2 toxin; the fumonisins B
1
, B
2
, and B
3
; agaric
acid; the ergot alkaloids; OTA; patulin; phomopsins;
sterigmatocystin, and zearalenone. Most of the limits
are set for human foods. Typically higher regulatory
levels are used for animal feed (101).
Risks associated with mycotoxins depend on both
hazard and exposure. Exposure throughout the
world is at different levels, because of different levels
of contamination and dietary habits in the various
parts of the world. Food and Drug Administration
(FDA) action levels for total AFs in food and feed are
presented in Table 1. European Union regulations for
AFs for human food and feed are presented in Table
2. EU regulations for OTs are showed in Table 3.
Mycotoxin Studies in Baby Foods and Infant
Formulas
There are several studies in literature on the levels
of several mycotoxins in infant formulas and baby
foods. These studies are largely based on AFs and
OTA and only few of them consider exposure to
other mycotoxins through baby foods.
In a study performed in Czechoslovakia using
enzyme-linked immunosorbent assay (ELISA)
technique, AFM1 was measured in 376 samples of
raw milk from farms in the area of a new dairy plant
producing milk baby foods. 87.8 % of the samples
contained no AFM1 (detection limit 0.025 mg/L) and
only 2 samples (0.5 %) possessed higher concentration
Table 1. FDA action levels for total aflatoxins in food and feed
Commodity AF (μg/kg)
All products, except milk, designated for humans 20
All other feedstuffs 20
Peanuts and Peanut products 20
Pistachio nuts 20
Milk 0.5 (for AFM1)
Foods 20
FABAD J. Pharm. Sci., 33, 51–66, 2008
57
than 0.1 mg/L, which represents the tolerance limit
for AFM1 in baby milk foods admitted in the country
(102). In another study performed in São Paulo,
Brazil AFM1 was surveyed in 300 samples of whole
milk powder consumed by infants at municipal
schools and nurseries. The analyses were performed
by ELISA. Results showed 11 % of the samples were
positive for AFM1 at levels of 0.10-1.00 ng/ml (mean:
0.27 ± 0.20 ng/ml) (103). In a study performed in
Turkey, the risk of exposure to aflatoxin in infants
fed by breast milk and formula was investigated. For
this purpose, AFB1 was determined in the serum of
both breast-fed and formula-fed infants. Serum AFB1
availability was significantly higher in the formula-
feeding (F) group than the breast-feeding (B) group
(42.8 vs 8.5 %). The AFB1 concentration in different
commercial formulas was also determined. AFB1 was
found in seven of the eight newly opened packages
of different brands of formula. The concentrations
showed a statistically significant increase at the 30
th
day after the opening of the packages. It was again
concluded that for infants, human milk was safer
than commercial formulas because of the lower
contamination risk of AF (104).
In an Italian study performed during 1995, 159
samples of milk, 97 samples of dry milk for
infant formula, and 114 samples of yogurt were
randomly collected in supermarkets and drug
stores in four large Italian cities and checked
for AFM1 by immunoaffinity column extraction
and high performance liquid chromatography
(HPLC). AFM1 was detected in 136 (86 %) of the
milk samples (in amounts ranging from <1 ng/L to
108.5 ng/L; mean level: 10.19 ng/L), in 81 (84 %) of
the dry milk samples (in amounts ranging from <1-
101.3 ng/kg; mean level: 21.77 ng/kg), and in 91 (80
%) of the yogurt samples (in amounts ranging from
<1 ng/L to 496.5 ng/L; mean level: 18.08 ng/L).
Altogether, only two samples of milk, two samples
of yogurt, and one sample of dry milk had levels
of AFM1 exceeding the Swiss legal limits, which
are the most restrictive limits in the world. AFM1
contamination levels in milk and yogurt samples
collected in the period of November to April were
four times as high as those in samples collected
in the period of May to October (105). In another
study performed by the same working group in
year 1996, 161 samples of milk, 92 samples of dry
milk for infant formula and 120 samples of yoghurt,
were randomly collected in supermarkets and drug
stores in four big Italian cities, and checked for
AFM1 by HPLC. AFM1 was detected in 125 (78 %)
of milk samples (ranging from < 1-23.5 ng/L; mean
Table 2. EU regulations for aflatoxins
Human food
AFB1
(μg/kg)
AFB1, B2, B3, B4
(μg/kg)
AFM1
(μg/kg)
Groundnuts, dried fruit and processed products thereof 2 4 –
Groundnuts subjected to sorting or phys. treating 8 15 –
As above but for nuts and dried fruits 5 10 –
Cereals (including maize) and processed products thereof 2 4 –
Milk – – 0.05
Table 3. EU regulations for ochratoxin.
Product OTA (μg/kg)
Raw cereal grains 5
All products derived from cereals intended for direct human consumption 3
Dried vine fruit (currants, raisins and sultanas) 10
Baby food 0.5
Erkekoğlu, Şahin, Baydar
58
level: 6.28 ng/L), in 49 (53 %) of dry milk samples
(ranging from <1-79.6 ng/kg; mean level: 32.2 ng/
kg) and in 73 (61 %) of yoghurt samples (ranging
from <1-32.1 ng/kg; mean level: 9.06 ng/kg). In
both of the studies, evaluating all of the samples
analyzed, the researchers concluded that during
1996, despite the widespread occurrence of AFM1,
mean contamination levels in dairy products sold
in Italy were not a serious human health hazard
(105).
In a Korean study, the occurrence of AFM1
in pasteurized milk and dairy products was
investigated by ELISA and HPLC. Among a total
of 180 samples collected the incidence of AFM1 in
pasteurized milk, infant formula, powdered milk
and yoghurt was 76, 85, 75, and 83 %, respectively,
with a mean concentration of 18, 46, 200, and 29
pg/g, respectively (106). In another study performed
in Kuwait, as a part of the program on monitoring
of environmental contaminants in food stuff in the
country, 54 samples of fresh full cream and skimmed
milk, powdered milk, yoghurt, and infant formulae
were analyzed for AFM1 by HPLC. Of the samples
analyzed, 28 % were contaminated with AFM1 with
6 % being above the maximum permitted limit of 0.2
mg/L (107).
A survey of AFs, OTA and patulin in a variety of
foods for infants and young children was carried out
by the Food Standards Agency between November
2003 and April 2004 in the U.K. Total 199 foods,
including breakfast/rusk products, baby rice,
savory products and desert/cereal bar/biscuits
were sampled. Of these, 169 were analyzed for AFs
and OTA. A further 14 products were analyzed for
patulin as well and 16 products, including apple-
based drinks and apple fruit products, were tested
for patulin only, as patulin is much more likely
to occur in these products compared with the
other mycotoxins studied. Mycotoxins were not
detectable in 90 % of the products analyzed. In
those samples where mycotoxins were detectable,
levels were very low and regulatory limits were
not exceeded in any of them. Data from the survey
were used to assess the exposure of infants to
mycotoxins and these do not raise a concern for
infant health (108). There are also several studies
performed on apple-based baby foods. In a study
performed in Tunisia, the researchers did not find
any patulin contamination in 21 infant fruit purees
(109). In an Italian study, of 10 apple-based baby
foods, two samples were contaminated with 17.7
and 13.1 mg/L and both were labeled as “organic
food” (110). In another study performed in Italy,
patulin was detected (<1 µg/kg) in only 3 of the
23 fruity baby food samples tested (homogenized
fruits, 11 conventional and 12 organic) (111).
In a survey performed in Canada on breakfast and
infant cereals for AFs B1, B2, G1 and G2, 349 breakfast
and infant cereal samples (rice-, soy-, barley-based
and mixed-grain infant cereals, corn-, wheat-, and
rice-based and mixed-grain breakfast cereals) were
collected at retail level across the country from 2002
to 2005. Results showed that 50 % of both breakfast
and infant cereals had detectable levels (limit of
detection = 0.002 ng/g) of AFB1. The levels found
varied from 0.002 to 1.00 ng/g for AFB1, from 0.002
to 0.14 ng/g for AFB2, from 0.008 to 0.27 ng/g for
AFG1, and from 0.008 to 0.048 ng/g for AFG2. Only
4 % of the breakfast cereals and 1 % of the infant
cereals had AFB1 levels exceeding 0.1 ng/g, which
is the European Union maximum limit for AFB1
in baby foods and processed cereal-based foods
for infants and young children (112). In another
Canadian study demonstrated on three hundred and
sixty-three samples of cereal-based infant foods, soy-
based cereals (which usually contain corn) exhibited
the highest incidences of deoxynivalenol (100%),
zearalenone (46%) and fumonisins (75%). Overall,
deoxynivalenol was the most frequently detected
mycotoxin--it was detected in 63% of samples
analyzed (113).
In another study performed in Russia, OTA content
in baby foods was determined. The analysis was
performed by immunoaffinity column clean-up and
HPLC. OTA was detected in 22.5 % of 40 samples
up to 1.2 mg/kg. Mean level was 0.15 and 0.31 mg/
kg. OTA level was higher in oat-based samples.
Calculations made on the basis of the obtained means
showed that the daily OTA dietary intake were up to
1.72 ng/kg. b.w. (114).
FABAD J. Pharm. Sci., 33, 51–66, 2008
59
Mycotoxin Studies in Baby Foods Performed in
Turkey
In a study conducted by Baydar et al., 63 infant
formulae, follow on formulae and baby foods
were randomly collected from pharmacies and
supermarkets in Ankara, Turkey. AFB1, AFM1, and
OTA levels were assessed by ELISA. AFB1, AFM1and
OTA levels were found in 87, 36.5 and 40 % of the
samples between 0.10-6.04 ppb, 0.06-0.32 ppb and
0.27-4.50 ppb, respectively (115). Another study
performed in Turkey on 24 cereal-based baby foods
using immunoaffinity column (IAC) clean-up and
HPLC determined that OTA was present in 17% of
cereal-based baby food samples. OTA levels ranged
from 0.122 to 0.374 ng/ml and the levels were much
lower below the limit recommended by European
Commission Regulation (116).
Recently, Gürbay et al. have indicated OTA levels in 75
Turkish mother breast milk samples ranging from 0.62
to 13 ng/L (117). Besides, the same group performed
another study to determine the levels of AFB1 and
AFM1 in breast milk. The level of AFM1 were in the
ranges of 60.90-299.99 ng/L, and AFB1 were in the
ranges of 94.50-4123.80 ng/L (118). Since there is no
limit value for AFM
1
and AFB
1
in mother’s breast
milk neither in Turkey nor European Union, making
a comparison of these results with limit values could
not be possible. However, when limit value of AFM
1
for animal milk (50 ng/l) accepted by Turkey and
European Union is considered, it has been shown that
all samples analyzed, contained AF M
1
above this limit.
Moreover, as there is not any limit for AFB
1
concerning
animal milk, it is not feasible to compare these
results of AFB
1
. On the other hand, The Joint FAO/
WHO Expert Committee on Food Additives (JEFCA)
does not establish a tolerable daily intake (TDI) for
aflatoxins, but strongly recommends that the level of
aflatoxin should be as low as possible (119). As a result,
the authors concluded that breast milk AFM
1
and B
1
levels determined in this study should be considered
seriously high and detrimental to human health.
Another study performed on human breast milk
and row cow’s milk using ELISA and HPLC,
demonstrated that AFM1 was present in 8 (13.1%) of
61 human breast milk samples examined (5.68±0.62
ng/L; ranged between 5.10- 6.90 ng/L) and 20 (33.3%)
of 60 raw cow’s milk samples (ranged between 5.40
-300.20 ng/L). Five (8.3%) of the positive raw cow’s
milk samples had AFM1 levels (153.52 ±100.60 ng/L;
ranged between 61.20-300.20 ng/L). The levels
in these samples were higher than the maximum
tolerance limit (0.05 ppb) stipulated by regulations in
Turkey and some other countries (120).
Regulations in Turkey
For baby foods and infant formulas, the
permissible levels for total aatoxin contamination
(AFB1+B2+G1+G2) is 2 ppb as indicated in
“Announcement for Aatoxin Control” by Turkish
Ministry of Agriculture, Forestry and Rural Affairs,
published in Turkish Republic Ofcial Paper on
May 2, 1990 (121). Turkish Ministry of Agriculture
and Rural Affairs set different limits for different
food stuff in 2002 and published “The Regulation
Stating a Change in the Regulation of the Turkish
Food Codex” in Turkish Republic Ofcial Paper No
24885, September 23, 2002. These regulations set by
Turkish Ministry of Agriculture and Rural Affairs are
summarized in Table 4.
Is The Mycotoxin Contamination In Baby Foods
A Treat?
The reaction of infants and young children differ from
that of adults against many drugs and toxins and, in
most cases, they are more susceptible. Furthermore,
infants and young children eat and drink more
relative to their size than adults. The fact that most
mycotoxins are toxic in very low concentrations and
they can be present in infant formulas and baby foods
as a result of contamination or bad storage, there is
the need for exible, reliable, accurate, inexpensive,
rapid and reproducible methods for detection and
quantication. Due to the varied structures of these
compounds, it is not possible to use one standard
technique to detect all mycotoxins, as each will
require a different method. What works well for
some molecules could be inappropriate for others
of similar properties or for the same molecule in a
different environment/matrix. As baby foods and
infant formulas have complex matrices, practical
requirements for high-sensitivity detection and
the need for a specialist laboratory setting create
Erkekoğlu, Şahin, Baydar
60
challenges for routine analysis. Therefore, depending
on the physical and chemical properties, procedures
have been developed around existing analytical
techniques, which offer exible and broad-based
methods of detecting compounds. Among other
methods, analytical liquid chromatography linked
with mass spectroscopy is gaining popularity (122).
WHO recommends exclusive breast-feeding for the
first six months of life. Breast milk remains the best
source of nutrition for infants, and mothers need to
be motivated to continue with it as long as possible.
However, when breast-feeding is not possible or
enough, many different infant formulae and baby
foods are available for infants and young children.
Preparing or buying safe and proper food for infants
and young children is essential for the health of the
child. Although the qualities of these products are
strictly regulated, contamination is inevitable. Most
mycotoxins are chemically stable so they tend to
survive in storage and processing, even when cooked
at quite high temperatures such as those reached
during bread baking or breakfast cereal production.
This makes it important to avoid the conditions
that lead to mycotoxin formation. Mycotoxins are
notoriously difficult to remove and the best method
of control is prevention (123).
Mycotoxin contamination is a very important issue as
the possible outcomes in the exposure to these toxins
may be the cause of serious problems experienced
in the rst years or later periods of life such as poor
growth, suppressed immune system, and cancer. An
accurate prediction of the possible health impact of
individual mycotoxins in foods for the vulnerable
group is difcult; possible additive and synergistic
effects of multiple mycotoxins make the task even
more complex and the long-term effects are beyond
foresight. Therefore, infant foods must be routinely
tested for the mycotoxins presence at every step of
manufacturing and marketing (116).Therefore, strict
law enforcement is required in each step of the
production of infant formulas/baby foods and the
public consciousness must be provided for the risks
Table 4. Maximum permissible levels for mycotoxins in Turkey.
Product
Total Aatoxin
(B1+B2+G1+G2)
(ppb)
AFM1
(ppb)
AFB1
(ppb)
OTA
(ppb)
Baby foods/infant formula (milk based) - 0.05 - -
Baby foods/infant formula 2 - 1 -
Spice 10 - 5 -
Milk 0.5 0.05 - -
Milk powder - 0.5 - -
Cheese - 0.25 - -
Agricultural products 20 - - -
Animal feed 50 - - -
Other foodstuff 10 - 5 -
Nuts, ground nuts and dried oily fruits, oily seeds,
dried fruits including g and grape, foodstuff prepared
from the procession of these
10 - 5 -
Cereals (including black wheat Fagopyrum) and all
products prepared using cereal
4 2 3
Processed cereal seproducts - - - 5
Dried grape - - - 10
Apple juice, fruit juices including apple juice, vinegars - - - 50
FABAD J. Pharm. Sci., 33, 51–66, 2008
61
of mycotoxins. Parents should be well-informed for
the right choice and for the use of the product they
bought (how to preserve, how to prepare etc) and
governments should pursuit the production of each
food produced for children.
Conclusion
It can be suggested that manufacturers of foods for
infants and young children should give an extreme
importance to mycotoxin content. The manufacturers,
pediatrician, health-care personnel and parents
should be provided with enough information
and training to minimize health hazards and to
form the public policies. In order to protect public
health, it is essential to keep contaminants at levels
toxicologically acceptable. Ultimately, surveillance
should be continuous, widespread and must be
conducted by the government and related ministries
as the quality of the end product depend on the
precise controlling at every step of the production
REFERENCES
1. Bryden WL. Mycotoxins in the food chain:
human health implications. Asia Pac J Clin Nutr
16:95-101, 2007.
2. Shephard GS. Aflatoxin analysis at the beginning
of the twenty-first century. Anal Bioanal Chem
395: 1215-1224, 2009.
3. Shephard GS. Determination of mycotoxins in
human foods. Chem Soc Rev 37: 2468-2477, 2008.
4. Dorner JW. Management and prevention of
mycotoxins in peanuts. Food Addit Contam Part
A Chem Anal Control Expo Risk Assess 25:203-208,
2008.
5. Turner NW, Subrahmanyam S, Piletsky SA.
Analytical methods for determination of
mycotoxins: a review. Anal Chim Acta 632: 168-
180, 2009.
6. Ardic M, Karakaya Y, Atasever M, Durmaz H.
Determination of aflatoxin B(1) levels in deep-
red ground pepper (isot) using immunoaffinity
column combined with ELISA. Food Chem Toxicol
46:1596-1599, 2008.
7. Korpinen EL. rence of aflatoxin in ground nuts,
some other nuts and industrial proteins imported
into Finland. Nord Hyg Tidskr 52:60-69, 1971.
8. Dorner JW. Biological control of aflatoxin
contamination in corn using a nontoxigenic
strain of Aspergillus flavus. J Food Prot 72:801-
804, 2009.
9. Dorner JW. Management and prevention of
mycotoxins in peanuts. Food Addit Contam Part
A Chem Anal Control Expo Risk Assess 25:203-208,
2008.
10. Karaca H, Nas S. Aflatoxins, patulin and
ergosterol contents of dried figs in Turkey. Food
Addit Contam 23: 502-508, 2006.
11. Prandini A, Tansini G, Sigolo S, Filippi L, Laporta
M, Piva G. On the occurrence of aflatoxin M1 in
milk and dairy products. Food Chem Toxicol 47:
984-991, 2009.
12. Hsieh D. Potential human health hazards of
mycotoxins. In Natori S, Hashimoto K, Ueno Y,
editors. Mycotoxins and phytotoxins. Third Joint
Food and Agriculture Organization/W.H.O./
United Nations E? Program International
Conference of Mycotoxins. The Netherlands
(Amsterdam): Elsevier; 1988. p. 69-80.
13. Guengerich FP, Johnson WW, Shimada T, Ueng
YF, Yamazaki H, Langouët S. Activation and
detoxication of aflatoxin B1. Mutat Res 402:121-
128, 1998.
14. Raj HG, Prasanna HR, Mage PN, Lotlikar
PD. Effect of purified rat and hamster hepatic
glutathione S-transferases (GST) on the
microsome mediated binding of aflatoxin B
1
to
DNA. Cancer Lett 33: 1-91, 1986.
15. Eaton DL, Groopman JD. The toxicology of
aflatoxins: human health, veterinary, and
agricultural significance. 1
st
ed. San Diego:
Academic Press; 1994.
16. Eaton DL, Gallagher EP. Mechanisms of aflatoxin
carcinogenesis. Annu Rev Pharmacol Toxicol 34:
135-172, 1994.
17. International Agency for Research on Cancer
(IARC). Monographs on the evaluation of the
carcinogenic risk of chemicals to man. Geneva:
World Health Organization, 1972.
18. Olivier M, Hussain SP, Caron de Fromentel C,
Hainaut P, Harris CC. TP53 mutation spectra
and load: a tool for generating hypotheses on
the etiology of cancer. IARC Sci Publ 157: 247-270,
2004.
Erkekoğlu, Şahin, Baydar
62
19. Van Vleet TR, Watterson TL, Klein PJ, Coulombe
RA Jr. Aflatoxin B1 alters the expression of p53 in
cytochrome P450-expressing human lung cells.
Toxicol Sci 89:399-407, 2006.
20. Farazi PA, DePinho RA. The genetic and
environmental basis of hepatocellular carcinoma.
Discov Med 6: 182-186, 2006.
21. Hengstler JG, Van der Burg B, Steinberg P, Oesch
F. Interspecies differences in cancer susceptibility
and toxicity. Drug Metab Rev 31:917-970, 1999.
22. Ciegler AM, Bennet JW. Mycotoxins and
mycotoxicoses. Bioscience 30:512, 1980.
23. Autrup H, Seremet T, Wakhisi J. Evidence for
human antibodies that recognize an aflatoxin
epitope in groups with high and low exposure to
aflatoxins. Arch Envir Health 45:31-34, 1990.
24. Nuntharatanapong N, Suramana T,
Chaemthavorn S, Zapuang K, Ritta E,
Semathong S, Chuamorn S, Niyomwan V,
Dusitsin N, Lohinavy O, Sinhaseni P. Increase
in tumor necrosing factor-alpha and a change
in lactate dehydrogenase isoeznyme pattern
in plasma of workers exposed to aflatoxin-
contaminated feeds. Arh Hig Rada Toksikol 52:
291-298, 2001.
25. Ranjan BG, Bhat NK, Avadhani NG. Preferential
attack of mitochondrial DNA by aflatoxin B1
during hepatocarcinogenesis. Science 215:73-75,
1982.
26. Shanks ET, Statkiewicz WR, Llwelleyn GC,
Dashek WV. Tissue distribution and hepatic
ultrastructural effects of aflatoxin B1 in Japanese
quail. Pol Arch Weter 26: 117-131, 1986.
27. Rainbow L, Maxwell SM, Hendrickse RG.
Ultrastructural changes in murine lymphocytes
induced by aflatoxin B1. Mycopathologia 125:33-
39, 1994.
28. Obasi SC. Effects of scopoletin and aflatoxin B1
on bovine hepatic mitochondrial respiratory
complexes, 2: a-ketoglutarate cytochrome c and
succinate cytochrome c reductases. Z Naturforsch
[C] 56:278-282, 2001.
29. Pasupathy K, Krishna M, Bhattacharya RK.
Alterations of phosphatidylinositol signal
pathway in hepatic mitochondria following
aflatoxin B1 administration. Indian J Exp Biol
47:876-880, 1999.
30. Sajan MP, Satav JG, Bhattacharya RK.
Alteration of energy-linked functions in rat
hepatic mitochondria following aflatoxin B1
administraton. J Bochem Toxicol 11:235-241,
1996.
31. Toskulkao C, Glinsukon T. Hepatic mitochondrial
function and lysosomal enzyme activity in
ethanol-potentiated aflatoxin B1 hepatotoxicity.
Toxicol Lett 52:179-90, 1990.
32. Reddy RV, Taylor MJ, Sharma RP. Studies of
immune function of CD-1 mice exposed to
aflatoxin B1. Toxicology 43:123-132, 1987.
33. Reddy RV, Sharma RP. Effects of aflatoxin B1 on
murine lymphocytic functions. Toxicology 54: 31-
44, 1989.
34. Hatori Y, Sharma RP, Warren RP. Resistance of
C57Bl/6 mice to immunosuppressive effects of
aflatoxin B1 and relationship with neuroendocrine
mechanisms. Immunopharmacology 22: 127-136,
1991.
35. Turner PC, Moore SE, Hall AJ, Prentice AM, Wild
CP. Modification of immune function through
exposure to dietary aflatoxin in Gambian
children. Environ. Health Perspect. 111: 217-220,
2003.
36. Jiang Y, Jolly PE, Ellis WO, Wang JS, Phillips TD,
Williams JH. Aflatoxin B1 albumin adduct levels
and cellular immune status in Ghanaians. Int.
Immunol. 17: 807-814, 2005.
37. Opinion of the Scıentıfıc Panel on Contamınants
in The Food Chain on a Request from the
Commission Related to The Potential Increase
of Consumer Health Risk by a Possible Increase
of the Existing Maximum Levels for Aflatoxıis in
Almonds, Hazelnuts and Pistachios and Derived
Products. Question N EFSA-Q-2006-174. EFSA
J.446,1 – 127, 2007. Available at: http://www.efsa.
europa.eu/en/efsajournal/doc/CONTAM%20_
op_ej446_aflatoxins_en.pdf?ssbinary. Last
accessed: 15 April 2010.
38. Kuiper-Goodman T. Uncertainties in the risk
assessment of three mycotoxins: aflatoxin,
ochratoxin, and zearalenone. Can J Physiol
Pharmacol 68:1017-1024, 1990.
39. European Mycotoxin Awareness Network.
Available at: Available from: URL: http://www.
mycotoxins.org/.
FABAD J. Pharm. Sci., 33, 51–66, 2008
63
40. Galvano F, Galofaro V, Ritieni A, Bognanno M, De
Angelis A, Galvano G. Survey of the occurrence
of aflatoxin M1 in dairy products marketed in
Italy: second year of observation. Food Addit
Contam 18: 644-666, 2001.
41. International Agency for Research on Cancer
(IARC). Some naturally occurring substances:
food items and constituents, heterocyclic
aromatic amines and mycotoxins. In: IARC
monographs on the evaluation of carcinogenic
risk to humans, Lyon: IARC Scientific
publications, p. 599, 1993.
42. Chelcheleh M, Allameh A. In vivo biotransfor-
mation of aflatoxin B1 and its interaction with
cellular macromolecules in neonatal rats. Mech
Ageing Dev 78:189-196, 1995.
43. Pietri A, 2003, Van Egmond H.P. Aflatoxin M1:
occurrence, toxicity, regulation. In: Van Egmond
H, editor. Mycotoxins in dairy products, London
and New York: Elsevier Applied Science; 1989. p.
11–55.
44. Scientific Committee on Food (SCF) Opinion on
Aflatoxins, Ochratoxin A, and Patulin expressed
on 23 September 1994 (94
th
Meeting of the SCF).
European Commission Report of the Scientific
Committee for Food (35
th
series), pp. 45–50, 1994.
45. Available from: URL: http://www.sciencedirect.
com/science?_ob=RedirectURL&_method
=extern ObjLink&_locator=url&_cdi=5036&_
plusSign=%2B&_targetURL=http%253A%252
F%252Feuropa.eu.int%252Fcomm%252Ffood%
252Ffs%252Fsc%252Fscf%252Freports%252Fs
cf_reports_35.pdf.
46. Barnes JM. Aflatoxin as health hazard. J Appl
Bacter 33: 285–298, 1970.
47. Lafont P, Siriwardana MG, DeBoer E.
Contamination of dairy products by fungal
metabolites. J Environ Pathol Toxicol Oncol 10: 99-
102, 1990.
48. Govaris A, Roussi V, Koidis PA, Botsoglou NA.
Distribution and stability of aflatoxin M1 during
production and storage of yoghurt. Food Addit
Contam 19: 1043-1050, 2002.
49. Van der Merwe KJ, Steyne PS, Fourie LF, Scott
DB, Theron JJ. Ochratoxin A, a toxic metabolite
produced by Aspergillus ochraceus Wilh. Nature
205: 1112-1113, 1965.
50. Marquardt RR, Frohlich AA. A review of recent
advances in understanding ochratoxicosis. J
Anim Sci 70: 3968-3988, 1992.
51. International Agency for Research on Cancer
(IARC). Ochratoxin A. In: IARC monographs on
the evaluation of carcinogenic risks to humans.
Some naturally occurring substances: food items
and constituents, heterocyclic aromatic amines
and mycotoxins. Geneva: International Agency
for Research on Cancer. Vol. 56, 1993, p. 489-521.
52. Størmer FC, Lea T. Effects of ochratoxin A
upon early and late events in human T-cell
proliferation. Toxicology 95: 45-50, 1995.
53. O’Brien E, Prietz A, Dietrich DR. Investigation of
the teratogenic potential of ochratoxin A and B
using the FETAX system. Birth Defects Res B Dev
Reprod Toxicol 74:417-423, 2005.
54. Biró K, Barna-Vetró I, Pécsi T, Szabó E, Winkler
G, Fink-Gremmels J, Solti L. Evaluation of
spermatological parameters in ochratoxin A--
challenged boars. Theriogenology 60: 199-207,
2003.
55. de Groene EM, Hassing IG, Blom MJ, Seinen W,
Fink-Gremmels J, Horbach GJ. Development of
human cytochrome P450-expressing cell lines:
application in mutagenicity testing of ochratoxin
A. Cancer Res 56: 299-304, 1996.
56. Creppy EE. Human ochratoxicosis. J Toxicol Toxin
Rev 18: 277-293, 1999.
57. Pfohl-Leszkowicz A, Manderville RA. Ochratoxin
A: An overview on toxicity and carcinogenicity
in animals and humans. Mol Nutr Food Res 51:
61–99, 2007.
58. Harris JP, Mantle PG. Biosynthesis of ochratoxins
by Aspergillus ochraceus. Phytochemistry 58: 709-
716, 2001.
59. Bunge I, Heller K, Roschenthaler R. Isolation and
purification of ochratoxin AZ Lebensm Unters
Forsch 168: 457-458, 1979.
60. McMasters DR, Vedani A. Ochratoxin binding to
phenylalanyl-tRNA synthetase: computational
approach to the mechanism of ochratoxicosis
and its antagonism. Med Chem 42:3075-3086,
1999.
61. Rahimtula AD, Bereziat JC, Bussacchini-Griot V,
Bartsch H. Lipid peroxidation as a possible cause
of ochratoxin A toxicity. Biochem Pharmacol
Erkekoğlu, Şahin, Baydar
64
37: 4469-4475, 19Woolf LI. The heterozygote
advantage in phenylketonuria. Am J Hum Genet
38: 773-775, 1986.
62. Schwartz GG. Hypothesis: does ochratoxin A
cause testicular cancer? Cancer Causes Control 13:
91-100, 2002.
63. US-NTP (United States-National Toxicology
Program), 1989. Toxicology and Carcinogenesis
Studies of Ochratoxin A (CAS No. 303–47–9) in
F344/N Rats (Gavage Studies). Technical Report
Series No 358. NTIS Publication No. PB90–
219478/AS. Research Triangle Park, NC and
Bethesda, MD: National Toxicology Program.
142 pp.
64. Opinion of the Scientific Panel on Contaminants
in the Food Chain on a Request from the
Commissoin Related to Ochratoxin A in Food.
Question N° EFSA-Q–2005–154. EFSA J 365:1
– 56, 2006. Available at: http://www.efsa.
europa.eu/en/scdocs/doc/contam_op_ej365_
ochratoxin_a_food_en.pdf. Last accessed: 15
April 2010.
65. FAO/WHO (Food and Agriculture Organisation/
World Health Organisation), 2001. Ochratoxin
A. In: Safety evaluation of certain mycotoxins
in food, Prepared by the 56
th
Meeting of the
Joint FAO/WHO Expert Committee on Food
Additives (JECFA). WHO Food Additives Series
47, pp 281–387. World Health Organisation,
Geneva, Switzerland.
66. FAO/WHO (Food and Agriculture
Organisation/World Health Organisation),
1996. Toxicological evaluation of certain food
additives and contaminants. Joint FAO/WHO
Expert Committee on Food Additives (JECFA).
WHO Food Additive Series 35. World Health
Organisation, Geneva, Switzerland.
67. International Agency for Research on Cancer
(IARC). Some naturally occuring and synthetic
food components, furocoumarins and ultraviolet
radiation. IARC Monographs on the Evaluation
of Carcinogenic Risk of Chemicals to Humans;
Lyon: IARC, Vol. 40, 1986; p.83-98.
68. World Health Organization (WHO). Biological
hazards. Chapter 2. Foodborne hazards. Pp.1-
12. Available from: URL: http://www.who.int/
foodsafety/ publications/ capacity /en/2.pdf
69. Food for thought. New food safety regulations/
guidelines. Vol 3, Issue 3, 2003. Available
at: http://www.doh.gov.za/department/
foodcontrol/newsletter/2003/december.html.
Last accessed: 15 April 2010.
70. Gökmen V, Acar J. Long-term survey of patulin
in apple juice concentrates produced in Turkey.
Food Addit Contam. 17:933-996, 2000.
71. Yurdun T, Omurtag GZ, Ersoy O. Incidence of
patulin in apple juices marketed in Turkey. J Food
Prot. 64:1851-1853, 2001.
72. Creppy EE. Update of survey, regulation and
toxic effects of mycotoxins in Europe. Toxicol Lett
127: 19-28, 2002.
73. The Merck Veterinary Manual. Fumonisin
Toxicosis. Available from: URL: http://
w w w. m e r c k v e t m a n u a l . c o m / m v m /
i n d e x . j s p ? c f i l e = h t m / b c / 2 1 2 2 1 1 .
htm&word=leukoencephalomalacia.
74. Rastogi S, Shukla Y, Paul BN, Chowdhuri
DK, Khanna SK, Das M. Protective effect of
Ocimum sanctum on 3-methylcholanthrene,
7,12-dimethylbenz(a)anthracene and aflatoxin
B1 induced skin tumorigenesis in mice. Toxicol
Appl Pharmacol 224:228-240, 2007.
75. United States National Toxicology Program),
1999. Toxicology and carcinogenesis studies of
fumonisin B1 (CAS no 116355-83-0) in F344/N
Rats and B6C3F Mice (Feed studies). NTP
Technical Report TR 496; NIH Publication No
99-3955. U.S. Department of Health and Human
Services, Research Triangle Park, North Carolina,
USA.
76. Cole RJ, Cox RH. Handbook of toxic fungal
metabolites. 1
st
ed. New York: Academic Press;
1981.
77. Seagrave, S. Yellow rain: a journey through
the terror of chemical warfare. In: Ueno Y.
editor. Trichothecenes: chemical, biological
and toxicological aspects. 1
st
ed. Amsterdam:
Elsevier; 1981. p. 81-9.
78. Joffe AZ. 1986. Fusarium species: their biology
and toxicology. New York: John Wiley and Sons,
1984.
79. Beasley VR. Toxigenic Fusarium species:
identity and mycotoxicology. Pennsylvania:The
Pennsylvania State University Press; 1989.
FABAD J. Pharm. Sci., 33, 51–66, 2008
65
80. Richard JL. Some major mycotoxins and their
mycotoxicoses--an overview. Int J Food Microbiol
119: 3-10, 2007.
81. Beasley VR, Lambert RJ. The apparently minimal
hazard posed to human consumers of products
from animals fed trichothecene-contaminated
grains. Vet Hum Toxicol 32Sl: 27-39, 1990.
82. Pestka J J, Bondy GS. Immunotoxic effects of
mycotoxins, In: Miller JD, Trenhold H. editor.
Mycotoxins in grain. Compounds other than
aflatoxins. St. Paul, Minn: Eagan; 1994. p. 339-58.
83. Rotter BA, Prelusky DB, Pestka J J. Toxicology
of deoxynivalenol (vomitoxin). J Toxicol Environ.
Health 48:1-34, 1996.
84. Rocha O, Ansari K, Doohan FM. Effects of
trichothecene mycotoxins on eukaryotic cells: a
review. Food Addit Contam 22: 369-78, 2005.
85. Pestka JJ, Smolinski AT. Deoxynivalenol:
toxicology and potential effects on humans. J
Toxicol Environ Health B Crit Rev 8: 39-69, 2005.
86. International Agency for Research on Cancer
(IARC). Monographs on the evaluation of
the carcinogenic risk of chemicals to humans:
some naturally occurring substances. Food
items and constituents, heterocyclic aromatic
amines and mycotoxins. Lyon: International
Agency for Research on Cancer, Vol. 56, 1993.
p. 397–444l.
87. Opinion of the Scientific Committee on Food
on Fusarium Toxins: Part 5: T2 toxin and HT2
toxin. 2001. p.1-25. Available from: URL: http://
ec.europa.eu/food/fs/sc/scf/out88_en.pdf.
88. CAST. Mycotoxins — risks in plant, animal
and human systems, Task Force Report No. 139,
Council for Agricultural Science and Technology,
Ames, Iowa 2003. p. 1–191.
89. Richard JL. Some major mycotoxins and their
mycotoxicoses--an overview. Int J Food Microbiol
119: 3-10, 2007.
90. Niyo KA, Richard JL, Tiffany LH. Effect of
T-2 mycotoxin ingestion on phagocytosis of
Aspergillus fumigatus conidia by rabbit alveolar
macrophages and on hematologic, serum
biochemical, and pathologic changes in rabbits,
Am J Vet Res 49 1766–1773, 1988.
91. Richard JL. Mycotoxins as immunomodulators
in animal systems. In: Bray G,. Ryan DH editors.
Mycotoxins, Cancer, and Health. Pennington
Center Nutrition Series, Louisiana: Louisiana
State University Press; 1991. p. 197–220.
92. Ler SG, Lee FK, Gopalakrishnakone P. Trends in
detection of warfare agents. Detection methods
for ricin, staphylococcal enterotoxin B and T-2
toxin. J Chromatogr A 1133:1-12, 2006.
93. Paterson RR. Fungi and fungal toxins as
weapons. Mycol Res; 110: 1003-1110, 2006.
94. Sirkka U, Nieminen SA, Ylitalo P. Acute
neurobehavioural toxicity of trichothecene T-2
toxin in the rat. Pharmacol Toxicol. 70:111-114, 1992.
95. European Commission Health and Consumer
Protection Directorate-General. Opinion of the
Scientific Commitee on Food on Fusarium Toxins,
Part 5: T-2 toxin and HT-2 Toxin. Adopted on May
2001. Available at: http: //ec.europa.eu/food/fs/
sc/scf/out88_en.pdf. Last accessed: 15 April 2005.
96. Fermentek Biotechnology Homepage.
Zearalenone. Available from: URL: http://www.
fermentek.co.il/zearalenone.htm.
97. Ding X, Lichti K, Staudinger JL. The mycoestrogen
zearalenone induces CYP3A through activation
of the pregnane X receptor. Toxicol Sci 91: 448-55,
2006.
98. Fermentek Biotechnology Homepage MSDS for
Zearalenone. Available from: URL: http://www.
fermentek.co.il/MSDS/Zearalenone-MSDS.htm.
99. International Programme On Chemical Safety
(INCHEM). World Health Organization. Safety
Evaluation Of Certain Food Additives And
Contaminants. Who Food Additives Series:
44. Available at: http://www.inchem.org/
documents/ jecfa/jecmono/v44jec14.htm. Last
accessed: 15 April 2005.
100. Direction générale de la santé. Bureau VS 3.
Avis du 8décembre1998 du Conseil supérieur
d’hygiène publique de France, relatif aux
mycotoxines dans l’alimentation, évaluation et
gestion du risque (section de l’alimentation et
de la nutrition. Available at: http://www.sante.
gouv.fr/adm/dagpb/bo/1999/99-11/a0110799.
htm. Last accessed: 15 April 2005.
101. van Egmond HP, Schothorst RC, Jonker MA.
Regulations relating to mycotoxins in food:
perspectives in a global and European context.
Anal Bioanal Chem 389:147-157, 2007.
Erkekoğlu, Şahin, Baydar
66
102. Fukal L, Brezina P. Determination of aflatoxin
M1 level in milk in the production of baby and
children’s food using immunoassay. Nahrung 35:
745-748, 1991.
103. de Oliveira CA, Germano PM. Evaluation of
enzyme-linked immunosorbent assay (ELISA)
in milk powder contaminated with known
concentrations of aflatoxin M1. Rev Saude Publica
30: 542-548, 1996.
104. Akşit S, Caglayan S, Yaprak I, Kansoy S. Aflatoxin:
is it a neglected threat for formula-fed infants?
Acta Paediatr Jpn 39: 34-36, 1997.
105. Galvano F, Galofaro V, de Angelis A, Galvano
M, Bognanno M, Galvano G. Survey of the
occurrence of aflatoxin M1 in dairy products
marketed in Italy. J Food Prot 61: 738-741, 1998.
106. Kim EK, Shon DH, Ryu D, Park JW, Hwang HJ,
Kim YB. Occurrence of aflatoxin M1 in Korean
dairy products determined by ELISA and HPLC.
Food Addit Contam 17: 59-64, 2000.
107. Srivastava VP, Bu-Abbas A, Alaa-Basuny, Al-
Johar W, Al-Mufti S, Siddiqui MK. Aflatoxin M1
contamination in commercial samples of milk
and dairy products in Kuwait. Food Addit Contam
18:993-997, 2001.
108. Survey of Baby Foods for Mycotoxins. FSIS 68/04.
p.1-45. Available from: URL: http://www.food.
gov.uk/multimedia/pdfs/fsis6804.pdf.
109. Mhadhbi H, Bouzouita N, Martel A, Zarrouk
H. Occurrence of mycotoxin patulin in apple-
based products marketed in Tunisia. J Food Prot.
70:2642-2645, 2007.
110. Ritieni A. Patulin in Italian commercial apple
products. J Agric Food Chem. 51:6086-6090, 2003.
111. Piemontese L, Solfrizzo M, Visconti A. Occurrence
of patulin in conventional and organic fruit
products in Italy and subsequent exposure as-
sessment. Food Addit Contam. 22:437-442, 2005.
112. Tam J, Mankotia M, Mably M, Pantazopoulos P,
Neil RJ, Calway P, Scott PM. Survey of breakfast
and infant cereals for aflatoxins B1, B2, G1 and
G2. Food Addit Contam 23: 693-699, 2006.
113. Lombaert GA, Pellaers P, Roscoe V, Mankotia
M, Neil R, Scott PM. Mycotoxins in infant cereal
foods from the Canadian retail market. Food
Addit Contam. 20:494-504, 2003.
114. Aksenov IV, Eller KI, Tutel’ian VA. Ochratoxin A
content in baby food. Vopr Pitan 75: 66-69, 2006.
115. Baydar T, Erkekoglu P, Sipahi H, Şahin G.
Aflatoxin B1, M1 and Ochratoxin A levels in
infant formulae and baby foods marketed in
Ankara, Turkey. J Food Drug Anal 15: 89-92,
2007.
116. Kabak B. Ochratoxin A in cereal-derived products
in Turkey: occurrence and exposure assessment.
Food Chem Toxicol 47:348-352, 2009.
117. Gürbay A, Girgin G, Sabuncuoğlu SA, Şahin G,
Yurdakök M, Yiğit Ş, Tekinalp G; Ochratoxin A:
is it present in breast milk samples obtained from
mothers from Ankara, Turkey? J Appl Toxicol, 30
(4): 329-333, 2010.
118. Gürbay A, Sabuncuoğlu SA, Girgin G, Şahin
G, Yiğit Ş, Yurdakök M, Tekinalp G; Exposure
of newborns to aflatoxin M1 and B1 from
mothers’breast milk in Ankara, Turkey; Food
Chem Toxicol, 48: 314-319, 2010.
119. Polychronaki N, C Turner P, Mykkänen H, Gong
Y, Amra H, Abdel-Wahhab M, El-Nezami H.
Determinants of aflatoxin M1 in breast milk in a
selected group of Egyptian mothers. Food Addit
Contam. 23: 700-708, 2006.
120. Keskin Y, Başkaya R, Karsli S, Yurdun T, Ozyaral
O. Detection of aflatoxin M1 in human breast
milk and raw cow’s milk in Istanbul, Turkey. J
Food Prot. 72: 885-889, 2009.
121. “Announcement for Aflatoxin Control”. Turkish
Ministry of Agriculture, Forestry and Rural
Affairs. Turkish Republic Official Paper, May 2,
1990.
122. World Health Organization. Evaluation of
certain food additives and contaminants: forty-
forth report of the Joint FAO/WHO Expert
Committee on Food Additives. Ochratoxin A
and Patulin. WHO Technical Report Series 859:
35–38, 1995.
123. World Health Organization. Toxicological
evaluation of certain food additives and
contaminants. Chapter Ochratoxin A. The
37
th
meeting of the Joint FAO/WHO Expert
Committee on Food Additives, WHO Geneva,
WHO Food Additives Series 28: 365–417, 1991.
