CHAPTER 30
Mycotoxins
A very large number of molds produce toxic
substances designated mycotoxins. Some are
mutagenic and carcinogenic, some display spe-
cific organ toxicity, and some are toxic by other
mechanisms. While the clear-cut toxicity of many
mycotoxins for humans has not been demon-
strated, the effect of these compounds on experi-
mental animals and their effect in in vitro assay
systems leaves little doubt about their real and
potential toxicity for humans. At least 14 myco-
toxins are carcinogens, with the aflatoxins being
the most potent.74 It is generally accepted that
about 93% of mutagenic compounds are carcino-
gens. With mycotoxins, microbial assay systems
reveal an 85% level of correlation between car-
cinogenicity and mutagenesis.74
Mycotoxins are produced as secondary me-
tabolites. The primary metabolites of fungi as
well as for other organisms are those compounds
that are essential for growth. Secondary metabo-
lites are formed during the end of the exponen-
tial growth phase and have no apparent signifi-
cance to the producing organism relative to
growth or metabolism. In general, it appears that
they are formed when large pools of primary
metabolic precursors such as amino acids, acetate,
pyruvate, and so on, accumulate. The synthesis
of mycotoxins represents one way the fungus
has of reducing the pool of metabolic precursors
that it no longer requires for metabolism.
For the detection of mycotoxins in foods, see
Chapter 11 and reference 56.
AFLATOXINS
Aflatoxins are clearly the most widely stud-
ied of all mycotoxins. Knowledge of their exis-
tence dates from 1960, when more than 100,000
turkey poults died in England after eating pea-
nut meal imported from Africa and South
America. From the poisonous feed were isolated
Aspergillus flavus, and a toxin produced by this
organism that was designated aflatoxin {Aspergil-
lus flavus toxin—A-fla-toxin). Studies on the
nature of the toxic substances revealed the fol-
lowing four components:
It was later determined that A parasiticus pro-
duces aflatoxins. Another Aspergillus species,
A. nominus, also produces aflatoxins.41 These
compounds are highly substituted coumarins, and
at least 18 closely related toxins are known. Afla-
toxin Bi (AFB1) is produced by all aflatoxin-
positive strains, and it is the most potent of all.
AFM1 is a hydroxylated product of AFB1 and
appears in milk, urine, and feces as a metabolic
product.27 AFL, AFLH1, AFQ1, and AFP1 are all
derived fromAFB1. AFB2 is the 2,3-dehydro form
OfAFB1, and AFG2 is the 2,3-dihydro form of
AFG1. The toxicity of the six most potent afla-
toxins decreases in the following order: B1 > M1
> G1 > B2 > M2 # G2.3 When viewed under ultra-
violet (UV) light, six of the toxins fluoresce as
noted:
BJ and B2—blue
G1—green
G2—green-blue
M1—blue-violet
M2—violet
They are polyketide secondary metabolites
whose carbon skeleton comes from acetate and
malonate.
The proposed partial pathway for AFB1 syn-
thesis is as follows: Acetate > norsolorinic acid
> averantin > averufanin > averufin > versiconal
hemiacetal acetate > versicolorin A >
sterigmatocystin> O-methylsterigmatocystin >
AFB1. Versicolorin A is the first in the pathway
to contain the essential C2—C3 double bond.
Requirements for Growth and
Toxin Production
No aflatoxins were produced by 25 isolates of
A. flavus/parasiticus on wort agar at 2°, 7°, 41°,
or 46°C within 8 days, and none was produced
under 7.5° or over 400C even under otherwise
favorable conditions.66 In another study employ-
ing Sabouraud's agar, maximal growth of
A. flavus and A, parasiticus occurred at 330C
when pH was 5.0 and water activity (aw) was
0.99.35 At 15°C, growth occurred at aw 0.95 but
not at 0.90, while at 27° and 33°C, slight growth
was observed at an aw of 0.85.The optimum tem-
perature for toxin production has been found by
many to be between 24° and 28°C. In one study,
maximal growth of A. parasiticus was 35°C, but
the highest level of toxin was produced at 25°C.69
The limiting moisture content for AFB1 and
AFB2On corn was 17.5% at a temperature of
24°C or higher, with up to 50 ng/g being pro-
duced.85 No toxin was produced at 13°C. Over-
all, toxin production has been observed over the
aw range of 0.93 to 0.98, with limiting values
variously reported as being 0.71 to 0.94.51 In
another study, no detectable quantities OfAFB1
were formed by A. parasiticus at aw values of 0.83
and 100C.55 The optimum temperature at aw 0.94
was 24°C (Figure 30-1). Growth without demon-
strable toxin appeared possible at ^ 0.83 on malt
agar-containing sucrose. It has been observed by
several investigators that rice supports the pro-
duction of high levels of aflatoxins at favorable
temperatures but none is produced at 50C on ei-
ther rice or cheddar cheese.59
Overall, the minimal and maximal parameters
that control growth and toxin production by these
eukaryotic organisms are not easy to define, in
part because of their diverse habitats in nature
and in part because of their eukaryotic status. It
seems clear that growth can occur without toxin
production.
AFG1 is produced at lower growth tempera-
tures than AFB1, and while some investigators
have found more AFB1 than AFG1 at around
aflatoxin B,
rate of growth
water activity temperature 0C
Figure 30-1 Growth and aflatoxin B1 production on
malt extract-glycerine agar at various water activity
values and temperatures. White columns: rate of
growth; black columns: average AFB1 production.
Source: From Northolt et al.,55 copyright © 1976 by
International Association of Milk, Food and Environ-
mental Sanitarians.
300C, others have found equal production. With
regard to A. fla vus and A. parasiticus, the former
generally produces only AFB and AFG.21 Aera-
tion favors aflatoxin production, and amounts of
2 mg/g can be produced on natural substrates
such as rice, corn, soybeans, and the like.21 Up
to 200 to 300 mg/L can be produced in broth
containing appropriate levels of Zn2+. The release
OfAFB1 by A. flavus appears to involve an en-
ergy-dependent transport system.
Production and Occurrence in Foods
With respect to production in foods, aflatoxin
has been demonstrated on fresh beef, ham, and
bacon inoculated with toxigenic cultures and
stored at 15°, 20°, and 300C8 and on country-
cured hams during aging when temperatures
approached 300C, but not at temperatures less
than 15°C or relative humidity (RH) over 75%.9
They have been found in a wide variety of foods,
including milk, beer, cocoa, raisins, soybean
meal, and so on (see below). In fermented sau-
sage at 25°C, 160 and 426 ppm of AFG1 were
produced in 10 and 18 days, respectively, and 10
times more AFG1 was found than B1.45 Aflatox-
ins have been produced in whole-rye and whole-
wheat breads, in tilsit cheese, and in apple juice
at 22°C. They have been demonstrated in the
upper layer of 3-month-old cheddar cheese held
at room temperature46 and on brick cheese at
12.8°C by A. parasiticus after 1 week but not for
A. flavus.69 AFB1 was found in 3 of 63 commer-
cial samples of peanut butter at levels less than
5 ppb.60 From a 5-year survey of around 500
samples of Virginia corn and wheat, aflatoxins
were detected in about 25% of corn samples for
every crop year, with 18% to 61% of samples
containing 20 ng/g or more and 5% to 29% con-
taining more than 100 ng/g.70 The average quan-
tity detected over the 5-year period was 21 to
137 ng/g (Table 30-1). Neither aflatoxins nor
zearalenone and ochratoxin A were detected in
any of the wheat samples. The 1988 drought led
to an increase in the amount of aflatoxin pro-
duced in corn in some midwestern states that
received less than 2 inches of rain in June and
July. About 30% of samples contained more than
20 ppb compared to 2 to 3 ppb levels during
normal rainfall.73
Cyclopiazonic acid is produced by some A.
flavus strains and it is thought to contribute to
the toxicity of aflatoxin. In an examination of
seven truckloads of corn, it was found in four at
levels of 25 to 250 ng/g.43 Also found in four of
five loads was deoxynivalenol (DON, vomitoxin)
at levels of 46 to 676 ng/g.
The effect of temperature cycling between 5°
and 25°C on production in rice and cheese has
been investigated. A. parasiticus produced more
toxin under cycling temperatures than at 15°, 18°,
or 25°C, while A. flavus produced less under
these conditions.59 On cheddar cheese, however,
less aflatoxin was produced than at 250C, and
these investigators noted that cheese is not a good
substrate for aflatoxin production if it is held
much below the optimum temperature for toxin
production.
Aflatoxin production has been demonstrated
to occur on an endless number of food products
in addition to those noted. Under optimal condi-
tions of growth, some toxin can be detected
within 24 hours-otherwise within 4 to 10 days.17
On peanuts, Hesseltine34 has made the following
observations:
• Growth and formation of aflatoxin occur
mostly during the curing of peanuts after
removal from soil.
• In a toxic lot of peanuts, only comparatively
few kernels contain toxin, and success in
detecting the toxin depends on collecting a
relatively large sample, such as 1 kg, for
assay.
• The toxin will vary greatly in amount even
within a single kernel.
• The two most important factors affecting
aflatoxin formation are moisture and tem-
perature.
The U.S. Food and Drug Administration (FDA)
has established allowable action levels of afla-
toxins in foods as follows: 20 ppb for food, feeds,
Brazil nuts, peanuts, peanut products, and pista-
chio nuts and 0.5 ppb for milk.42 A committee of
Table 30-1 Aflatoxin Levels in Dent Corn Grown in Virginia, 1976-1980

Collected at Harvest by Statistical

Collected from Trucks by Federal Grain Inspection Service (FGIS) Reporting Service (SRS)
1976 1977 1978 1979 1980 1978 1979
Total Aflatoxin, No. of No. of No. of No. of No. of No. of No. of
ng/g Samples (0Zo) Samples (0Zo) Samples (0Zo) Samples (0Zo) Samples (0Zo) Samples (°/o) Samples (0Zo)

ND* 11 (63) 52 (51) 63 (64) 81 (71) 18 (18) 79 (88) 93 (79)

<20 13 (10) 17 (17) 10 (10) 13 (11) 20 (20) 2 (2) 9 (8)
20-100 21 (17) 18 (18) 21 (21) 8 (7) 32 (32) 5 (6) 7 (6)
101-500 9 (7) 10 (10) 5 (5) 10 (9) 26 (26) 4 (4) 7 (6)
501-1,000 1 (1) (1) 2 (2)
>1,000 2 (2) 3 (3) 2 (2) 1 (1)
Total 123 101 99 114 99 90 117

%lncidence 37 49 36 29 82 12 21
% >20 ng/g 27 32 26 18 61 10 13
% >100 ng/g 10 14 5 11 29 4 7
Average level 48 91 21 34 137 13 36
(ng/g), all
samples
Average level 130 187 58 118 167 110 176
(ng/g),
positive
samples

*ND = Not detected.

Source: Shotwell and Hesseltine;70 copyright © 1983 by Association of Official Analytical Chemists.
the Codex Alimentarius Commission has recom-
mended the following maximum levels of my-
cotoxins in specific foods: 15 ug/kg of aflatox-
ins in peanuts for further processing; 0.05 jug/kg
of aflatoxin M1 in milk; 50 ug/kg of patulin in
apple juice and apple juice ingredients in other
beverages; and 5 ug/kg of ochratoxin A in cere-
als and cereal products.53
Relative Toxicity and Mode of Action
For the expression of mutagenicity, mamma-
lian metabolizing systems are essential for afla-
toxins, especially AFB1. Also essential is their
binding with nucleic acids, especially DNA.
While nuclear DNA is normally affected, AFB1
has been shown to bind covalently to liver mito-
chondrial DNA preferentially to nuclear DNA.54
Cellular macromolecules other than nucleic ac-
ids are possible sites for aflatoxins. The site of
the aflatoxin molecule responsible for mutage-
nicity is the C2—C3 double bond in the dihydro-
furofuran moiety. Its reduction to the 2,3-dihydro
(AFB2) form reduces mutagenicity by 200- to
500-fold.74 Following binding to DNA, point
mutations are the predominant genetic lesions
induced by aflatoxins, although frameshift mu-
tations are known to occur. The mutagenesis of
AFB1 has been shown to be potentiated twofold
by butylated hydroxyanisole (BHA) and buty-
lated hydroxytoluene (BHT) and much less by
propyl gallate employing the Ames assay, but
whether potentiation occurs in animal systems
is unclear.68
The LD50 OfAFB1 for rats by the oral route is
1.2 mg/kg, and 1.5 to 2.0 mg/kg for AFG1.11 The
relative susceptibility of various animal species
to aflatoxins is presented in Table 30-2. Young
ducklings and young trout are among the most
sensitive, followed by rats and other species.
Most species of susceptible animals die within 3
days after administration of toxins and show
gross liver damage, which, upon postmortem
examination, reveals the aflatoxins to be hepa-
tocarcinogens.89 The toxicity is higher for young

Table 30-2 Comparative Lethality of Single Doses of Aflatoxin B1

Animal Age (or Weight) Sex Route LD50 (mg/kg)

Duckling 1 day M PO 0.37
1 day M PO 0.56
Rat 1 day M-F PO 1.0
21 days M PO 5.5
21 days F PO 7.4
100g M PO 7.2
100g M IP 6.0
15Og F PO 17.9
Hamster 30 days M PO 10.2
Guinea pig Adult M IP ca. 1
Rabbit Weanling M-F IP ca. 0.5
Dog Adult M-F IP ca. 1
Adult M-F PO ca. 0.5
Trout 100g M-F PO ca. 0.5

Note: PO = oral; IP = intraperitoneal.

Source: Wogan.89
animals and males than for older animals and
females, and the toxic effects are enhanced by
low protein or cirrhogenic diets.
Circumstantial evidence suggests that aflatox-
ins are carcinogenic to humans. Among condi-
tions believed to result from aflatoxins is the
EFDV syndrome of Thailand, Reye's syndrome
of Thailand and New Zealand,1112 and an acute
hepatoma condition in a Ugandan child. In the
last a fatal case of acute hepatic disease revealed
histological changes in the liver identical to those
observed in monkeys treated with aflatoxins, and
an aflatoxin etiology was strongly suggested by
the findings.67 Two researchers who worked with
purified aflatoxin developed colon carcinoma.23
On the other hand, it has been noted that no my-
cotoxin has been linked with a specific cancer
in humans in the absence of chronic infection
with hepatitis B virus.77 Although some mycotox-
ins are extremely toxic to the young of many
animal species, the view exists that their toxicity
for humans is overstated.
Degradation
AFB1 and AFB2 can be reduced in corn by bi-
sulfite. When dried figs were spiked with 250
ppb OfAFB1 and subjected to several treatments,
1% sodium bisulfite effected a 28,2% reduction
in 72 hours; 0.2% H2O2 (added 10 minutes be-
fore sodium bisulfite) effected a 65.5% reduc-
tion; heating at 45° to 650C for 1 hour effected a
68.4% reduction; and ultraviolet (UV) radiation
effected a 45.7% reduction.2 Aflatoxin-contami-
nated cottonseed treated with ammonia and fed
to cows led to lower levels OfAFB1 and AFM1
in milk than nontreated product.36 When yellow
dent corn naturally contaminated with 1,600 ppm
aflatoxin was treated with 3% NaOH at 1000C
for 4 minutes, further processed, and fried, 99%
of the aflatoxin was destroyed.13
ALTERNARIA TOXINS
Several species of Alternaria (including A.
citri, A. alternata, A. solani, and A. tenuissima)
produce toxic substances that have been found
in apples, tomatoes, blueberries, grains, and other
foods. 7576 The toxins produced include
alternariol, alternariol monomethyl ether,
altenuene, tenuazonic acid, and altertoxin-I.75 On
slices of apples, tomatoes, or crushed blueber-
ries incubated for 21 days at 210C, several Alter-
naria produced each of the toxins noted at lev-
els up to 137 mg/100 g.75 In another study,
tenuazonic acid was the main toxin produced in
tomatoes, with levels as high as 13.9 mg/100 g;
on oranges and lemons, A. citri produced
tenuazonic acid, alternariol, and alternariol
monomethyl ether at a mean concentration of
1.15 to 2.66 mg/100 g.76 The fruits were incu-
bated at room temperature for 21 to 28 days.
In a study of 150 sunflower seed samples in
Argentina, 85% contained alternariol (mean of
187 fig/kg), 47% contained alternariol
monomethyl ether (mean of 194 ug/kg), and 65%
contained tenuazonic acid (mean of 6,692 jig/
kg).18 Following fermentation for 28 days by A.
alternata and separation into oil and meal, no
alternariol, 1.6 to 2.3% of tenuazonic, and 44 to
45% alternariol monomethyl ether were found
in oil, but none of these toxins were in the meal.18
An A. alternata strain produced stemphyltoxin
III, which was mutagenic by the Ames assay22
More information on the alternaria toxins can
be found in reference 16.
CITRININ
The citrinin mycotoxin is produced by Peni-
cillium citrinum, R vindication, and other fungi.
It has been recovered from polished rice, moldy
bread, country-cured hams, wheat, oats, rye, and
other similar products. Under long-wave UV
light, it fluoresces lemon yellow. It is a known
carcinogen. Of seven strains of R viridicatum
Citrinin
recovered from country-cured hams, all pro-
duced citrinin in potato dextrose broth and on
country-cured hams in 14 days at 200C to 300C
but not at 100C.90 Growth was found to be poor
at 100C.
Citrinin was identified from moldy foods ex-
amined in Germany, and it along with some other
mycotoxins could be produced in a synthetic
mediun.44
While citrinin-producing organisms are found
on cocoa and coffee beans, this mycotoxin as well
as others is not found to the extent of growth.
The apparent reason is the inhibition of citrinin
in P. citrinum by caffeine. The inhibition of citri-
nin appears to be rather specific, since only a
small decrease in growth of the organisms occurs.5
OCHRATOXINS
The ochratoxins consist of a group of at least
seven structurally related secondary metabolites
of which ochratoxin A (OA) is the best known
and the most toxic. OB is dechlorinated OA and
along with OC, it may not occur naturally. OA is
produced by a large number of storage fungi,
including^. ochraceus,A. alliaceus,A. ostianus,
A. mellus, and other species of aspergilli. Among
penicillia that produce OA are P. viridicatum,
R cyclopium, P variable, and others.
OA is produced maximally at around 300C and
aw 0.95.4 The minimum aw supporting OA pro-
duction by A. ochraceus at 300C in poultry feed
is 0.85.4 Its oral LD50 in rats is 20 to 22 mg/kg,
and it is both hepatotoxic and nephrotoxic.
Ochratoxin A
This mycotoxin has been found in corn, dried
beans, cocoa beans, soybeans, oats, barley, cit-
rus fruits, Brazil nuts, moldy tobacco, country-
cured hams, peanuts, coffee beans, and other
similar products. Two strains of A. ochraceus iso-
lated from country-cured hams produced OA and
OB on rice, defatted peanut meal, and when in-
oculated into country-cured hams.28 Two-thirds
of the toxin penetrated to a distance of 0.5 cm
after 21 days, with the other one-third located in
the mycelial mat. Of six strains of/? viridicatum
recovered from country-cured hams, none pro-
duced ochratoxins. From a study of four chemi-
cal inhibitors of both growth and OA production
by two OA producers at pH 4.5, the results were
potassium sorbate > sodium propionate> methyl
paraben > sodium bisulfite; while at a pH of 5.5,
the most effective two were methyl paraben and
potassium sorbate.83 Like most other mycotox-
ins, OA is heat stable. In one study, the highest
rate of destruction achieved by cooking faba
beans was 20%, and the investigators concluded
that OA could not be destroyed by normal cook-
ing procedures.26 Under UV light, OA fluoresces
greenish, while OB emits blue fluorescence. It
induces abnormal mitosis in monkey kidney
cells.
PATULIN
Patulin (clavicin, expansin) is produced by a
large number of penicillia, including P.
claviforme, P expansum, R patulum; by some
aspergilli (A. clavatus, A. terreus, and others);
and by Byssochlamys nivea and B. fulva.21
Patulin
Its biological properties are similar to those
of penicillic acid. Some patulin-producing fungi
can produce the compound below 2°C.3 This
mycotoxin has been found in moldy bread, sau-
sage, fruits (including bananas, pears, pine-
apples, grapes, and peaches), apple juice, cider,
and other products. In apple juice, levels as high
as 440 jug/L have been found, and in cider, levels
up to 45 ppm.
 Along with citrinin and ochratoxin A, it was iden-
tified from moldy foods examined in Germany.44
Minimum aw for growth of P expansum and/?
patulum has been reported to be 0.83 and 0.81,
respectively. In potato dextrose broth incubated
at 12°C, patulin was produced after 10 days by
P. patulum and/? roquefortii, with the former or-
ganism producing up to 1,033 ppm.7 Patulin was
produced in apple juice also at 12°C by B. nivea,
but the highest concentration was attained after
20 days at 210C after a 9-day lag.62 The next high-
est amount was produced at 300C, with much
less at 37°C. These investigators confirmed that
patulin production is favored at temperatures
below the growth optimum, as was previously
found by Sommer et al.72 The latter investigators
used/? expansum and found production over the
range 5° to -20 0 C, with only small amounts pro-
duced at 300C. In five commercial samples in
Georgia, patulin levels from 244 to 3,993 jug/L
were found, with a mean of 1,902 jig/L.88 The
overall incidence of patulin in apple juice has
been reviewed.33 Atmospheres of CO2 and N2 re-
duced production compared to that in air. To in-
hibit production, SO2 was found more effective
than potassium sorbate or sodium benzoate.62
The LD50 for patulin in rats by the subcutane-
ous route is 15 to 25 mg/kg, and it induces sub-
cutaneous sarcomas in some animals. Both patu-
lin and penicillic acid bind to —SH and —NH2
groups, forming covalently linked adducts that
appear to abate their toxicities. Patulin causes
chromosomal aberrations in animal and plant
cells and is a carcinogen.
PENICILLIC ACID
This mycotoxin has biological properties simi-
lar to patulin. It is produced by a large number
of fungi, including many penicillia (P
puberulum, for example) as well as members of
the A. ochraceus group. One of the best produc-
ers is P cyclopium. It has been found in corn,
beans, and other field crops and has been
produced experimentally on Swiss cheese. Its
LD50 in mice by subcutaneous route is 100 to
300 mg/kg, and it is a proved carcinogen.
Penicillic acid
Of 346 penicillia cultures isolated from salami,
about 10% produced penicillic acid in liquid
culture media, but 5 that were inoculated into
sausage failed to produce toxin after 70 days.19
In another study, some 183 molds were isolated
from Swiss cheese; 87% were penicillia, 93% of
which were able to grow at 50C. Thirty-five per-
cent of penicillia extracts were toxic to chick
embryos, and from 5.5% of the toxic mixtures
were recovered penicillic acid as well as patulin
and aflatoxins.6 Penicillic acid was produced at
5°C in 6 weeks by 4 of 33 fungal strains.
STERIGMATOCYSTIN
These mycotoxins are structurally and biologi-
cally related to the aflatoxins, and like the latter,
they cause hepatocarcinogenic activity in ani-
mals. At least eight derivatives are known.
Among the organisms that produce them are
Aspergillus versicolor, A. nidulans,A. rugulosus,
and others. The LD50 for rats by intraperitoneal
injection is 60 to 65 mg/kg. Under UV light, the
toxin fluoresces dark brick-red. Although not
often found in natural products, they have been
found in wheat, oats, Dutch cheese, and coffee
beans. While related to the aflatoxins, they are
not as potent. They act by inhibiting DNA
synthesis.
FUMONISINS
The fumonisins are produced by Fusarium
spp. on corn and other grains, and certain dis-
eases of humans and animals are associated with
the consumption of grains and grain products that
contain high levels of these molds.
The species demonstrated to produce
fumonisins include E anthophilum, E dlamini,
F. napiforme, F. nygami, F. moniforme, and
Eproliferation.52 The latter species produces large
quantities. F. moniliforme (formerly E verti-
cillioides; Gibberella fujikuroi) was the first to
be associated with the mycotoxin and it is the
best studied of the three. The prevalence of
E moniliforme is significantly higher in corn
from areas where a high rate of human esoph-
ageal cancer occurred than in low esophageal
cancer rate areas.48
There are at least seven fumonisins, four Bs
and at least three As: FB1, FB2, FB3, FB4, FA1,
FA2, and FA3. The major ones are FB1-FB3, and
the others are considered to be minor and less
well characterized. Of the three major toxins, FB1
(also designated macrofusine) is produced in the
largest quantities by producing strains. For ex-
ample, among nine strains of E moniliforme, the
range of FB!produced on autoclaved corn was
960 to 2,350 ug/g while for FB2 the range was
120 to 320 ^ig/g.63
Fusarin C is produced by E moniliforme but
apparently is not involved in hepatocarcinogenic
activity.31 It is mutagenic in the Ames test but
only after liver fraction activation.87 In a culture
medium, more was produced at pH < 6.0 than
above, and the highest yields were achieved be-
tween days 2 and 6 at around 28°C.29 Corn iso-
lates were shown to produce about 19 to 332 ug/g
when grown on corn.29
Growth and Production
In regard to optimum growth temperature and
pH, the maximum yield of FB1 by a strain of E
moniliforme in a corn culture occurred in 13
weeks at 200C with a yield of 17.9 g/kg dry
weight.1 The higher growth rate of the fungus
occurred at 25°C, not 200C, and the stationary
phase was reached in 4 to 6 weeks at either tem-
perature.1 In the same study, FB1 production com-
menced after 2 weeks of active growth and de-
creased after 13 weeks. Overall, the optimum
time and temperature for FB1 production was 7
weeks at 25°C. Good growth by anE moniliforme
strain at 25 and 300C over the pH range of 3 to
9.5 has been demonstrated.86 Little growth oc-
curred at 37°C over the same pH range. Culture
media were used with acidic pH values adjusted
with phosphoric acid.86 Neither E moniliforme
nor E proliferatum produces much toxin at aw
0.925.50 For one strain of the former grown on
sterile corn for 6 weeks at 25°C, the ppm OfFB1
produced were 6.8, 14.4, 93.6, and 102.6 at the
respective aw values of 0.925, 0.944, 0.956, and
0.968.50
The preservative compounds benzoic acid,
BHA, and carvacrol have been shown to inhibit
or retard the mycelial growth of a number of
Fusarium spp., with benzoic acid being the most
effective followed by carvacrol and BHA.82 The
simultaneous effect on fumonisin production is
unclear.
Prevalence in Corn and Feeds
It has been observed since the mid-1980s that
leukoencephalomalacia (LEM) in horses, pulmo-
nary edema (PE) of porcines, and esophageal
cancer (EC) in humans occur in areas of the world
where high levels of fumonisins are found in
grain-based foods.91 For example, the highest rate
of human EC in southern Africa occurs in the
Transkei where high levels of FB1 and FB2 are
found in corn. Concomitant with the occurrence
of the fumonisins is the presence of Fusarium
spp., especially E moniliforme. The incidence of
fumonisin FB1 in a high-risk county in China was
about two times higher than in a low-risk area,
although the differences were not statistically
significant.91 Trichothecenes (mainly deoxyni-
valenol) in addition to fumonisins were found in
corn from the high-risk area.
The incidence and prevalence OfFB1 in corn
and some corn products in six countries are sum-
marized in Table 30-3. The highest levels found
were in corn from an area in the Transkei, South
Africa, where EC occurred at high rates. The
range for these six samples was 3,020 to 117,520
ng/g with a mean of 53,740 ng/g.79 These levels
exceeded those found in 12 samples of mold corn
from the same general area, where the mean was
23,900 ng/g.61 Overall, FB1 was found at lower
Table 30-3 Incidence and Prevalence of Fumonisin B1 In Corn and Corn Products from Several
Countries

Samples Fumonisin Mean,

Products Country Pos./Total Range, ng ng/g Reference

Corn grits Switzerland 34/55 0-790 260 58

Corn grits South Africa 10/18 0-190 125 80
Corn grits U.S.A. 10/10 105-2,545 601 80
Corn meal/muffin mix U.S.A. 10/17 <200-15,600 57
Corn meal Switzerland 2/7 0-110 85 58
Corn meal South Africa 46/52 0-475 138 80
Corn meal U.S.A. 15/16 0-2,790 1,048 80
Corn meal Canada 1/2 0-50 50 80
Corn meal Egypt 2/2 1,780-2,980 2,380 80
Corn meal Peru 1/2 0-660 660 80
Corn* Charleston, SC 111 105-1,915 635 80
Corn* Transkei, S.Af. 6/6 3,020-117,520 53,740 80
Corn (good)* Transkei, S.Af. 12/12 50-7,900 1,600 61
Corn (good)+ Transkei, S.Af. 2/12 0-550 375 61
Corn (moldy)1 Transkei, S.Af. 12/12 3,450-46,900 23,900 61
Corn (moldy)t Transkei, S.Af. 11/11 450-18,900 6,520 61
White corn meal U.S.A. (MD) 3,500-7,450 14
Yellow corn meal U.S.A. (MD) 500-4,750 14
Tortilla, white U.S.A. (MD) 200-400 14
Yellow corn meal U.S.A. (AZ) 450-650 14
Yellow corn meal U.S.A. (NE) 500-2,500 14
Tortilla, white U.S.A. (AZ) 250-1,450 14
Tortilla, white U.S.A. (NE) 200-550 14
Maize + meal Botswana 28/33 20-1,270 247 [jg/kg 71
Maize Kenya 92/197 110-12,000 670 38

*From areas of respective countries where human esophageal cancer was high.
1
FrOm areas where esophageal cancer was low.

levels in corn grits, while in corn meal levels
tended to be higher (Table 30-3).
Feed samples from 11 U.S. states were exam-
ined for FB1.63 Of the 83 equine feeds that were
associated with equine LEM, 75% contained
>10 ug/g with a range of < 1.0 to 126 ug/g. Of
the 42 associated with porcine PE syndrome,
71% contained >10 u/g with a range of < 1.0 to
330 (xg/g. On the other hand, all 51 samples of
nonproblem feeds had <9 ug/g of FBi, with 94%
of the 51 being <6 ug/g.63 Of 71 retail samples
of corn-based and other grain products in Michi-
gan, 11 contained fumonisins including 10 of 17
corn-based products.57 The highest level OfFB1
found using ELISA was 15.6 ug/g in blue corn
meal.57 Based on literature reports OfFB1 in maize,
it has been estimated that people in the Netherlands
may be exposed to an intake of 1,000 ng/d.24
When Fusarium-contaminated corn that was
associated with outbreaks of mycotoxicosis in
various animals in Brazil was examined for FB1
and FB2, 20 of 21 samples revealed FB1 levels
that ranged from 200 to 38,500 ng/g, and 18/21
had an FB2 range of 100 to 12,000 ng/g.78 Ex-
cept for one isolate from this corn, all were
acutely toxic to ducklings. In a 1996-1997 study
of fumonisins in Spanish beers, 14 were positive
at levels from 4.76 ng/mL to 85.53 ng/mL.84
Physical/Chemical Properties of
FB1 and FB2
The chemical structure OfFB1 and FB2 is indi-
cated below.25 The two differ only by FB1 having
an —OH group in lieu of an H on carbon 10.
These toxins differ from most others in this chap-
ter in two ways: they do not possess cyclic or
ring groups, and they are water soluble. On the
other hand, they are heat-stable, as are many other
mycotoxins. In one study, lyophilized culture
materials containing FB1 were boiled for 30 min-
utes and then oven dried at 600C for 24 hours
without loss of toxic activity.l
In another study, the thermal stability of these
toxins at a level of 5 ug/g OfFB1 in processed
corn products was assessed.15 No significant loss
was found upon baking at 2040C for 30 min-
utes. Almost complete loss occurred upon roast-
ing corn meal samples at 218°C for 15 minutes.
Significant but not total reduction was noted in
cornbread at 232°C for 20 minutes. In regard to
the thermal stability in canned foods, 5 ug/g were
added to canned foods and then recanned. No
significant loss occurred in creamed corn for
infants and canned dog food, but significant re-
ductions occurred in cream-style corn and
whole-kernal corn, although it was not elimi-
nated.15 Overall, roasting was more effective than
baking.
Pathology
In experimental animals, the liver is the pri-
mary target of FB1. In a study using rats over a
26-month period, all animals that either died or
were killed after 18 months had micro- and
macronodular cirrhosis and large expansile nod-
ules of cholangiofibrosis at the hilus of the liver.30
(Cholangiof ibrosis is considered to be a precur-
sor lesion for cholangiocarcinoma in rats.) Of
15 rats that died or were killed between 18 and
26 months, 66% developed primary hepatocel-
lular carcinoma. Some involvement of the kid-
neys occurred but only toward the end of this
study. No esophageal lesions were noted in test
animals, and no neoplastic changes were noted
in the 25 controls.30 The hepatocarcinogenic ac-
tivity OfFB1 in rats was demonstrated by adding
50,000 ng/g in food rations over a 26-month pe-
riod.30 In an earlier study, FB1 was shown to pos-
sess cancer-promoting activity by its capacity to
elevate 7-glutamyltranspeptidase activity in rats.1
Leukoencephalomalacia (LEM) was repro-
duced in a horse by the intravenous (i.v.) injec-
tion of seven daily doses of FB1 at a level of
0.125 mg/g live mass spread over 10 days (see
reference 81). LEM was produced in two horses
via the oral administration of FB1 at a level of
[1] R = OH
[2] R = H
fumonisins B 1 [1] and B2 [2].
1.25-4 mg/g body weight, and symptoms oc-
curred in around 25 days (see reference 81).
Pulmonary edema was produced in a pig after
daily injections of 0.4 mg FB1Zg body weight for
4 days.81 The prevalence of human esophageal
cancer in the Transkei, South Africa, is statisti-
cally correlated with high levels OfFB1 and FB2
in corn.80
SAMBUTOXIN
The sambutoxin mycotoxin was first reported
in 1994,39 and its structure is shown below. It is
associated with dry-rotted potatoes and is pro-
duced primarily by strains of Fusarium
sambucinum and F. oxysporum. Of 13 Fusarium
species examined, about 90% of strains of the
two species noted produced this toxin. From rot-
ten potato samples in Korea, 9 of 21 contained
15.8 to 78.1 ng/g of sambutoxin with a mean of
49.2 ng/g.40 Using wheat media, levels of 1.1 to
101 |ig/g of sambutoxin were produced. The toxin
was found in potatoes from parts of Iran that had
a high incidence of esophageal cancer.39
Sambutoxin causes hemorrhage in the stom-
ach and intestines of rats, and the animals refuse
feed and lose weight.39 Rats died within 4 days
when their diets contained 0.1% sambutoxin. It
is toxic to chick embryos with an LD50 of 29.6
jig/egg.39
ZEARALENONE
There are at least five naturally occurring
zearalenones, and they are produced by Fusarium
spp., mainly Fgraminearum (formerly Froseum,
= Gibberella zeae) and E tricinctum. Associated
with corn, these organisms invade field corn at
the silking stage, especially during heavy rain-
fall. If the moisture levels remain high enough
following harvesting, the fungi grow and pro-
duce toxin. Other crops, such as wheat, oats,
barley, and sesame, may be affected in addition
to corn.
The toxins fluoresce blue-green under long-
Zearalenone
wave UV and greenish under short-wave UV
They possess estrogenic properties and promote
estrus in mice and hyperestrogenism in swine.
While they are nonmutagenic in the Ames assay,
they produce a positive response in the Bacillus
subtilis Rec assay.74
CONTROL OF PRODUCTION
A number of organisms, especially other fungi,
have been shown to control the growth of toxi-
genic fungi and to inhibit toxin production (for
reviews, see references 32 and 65). Among the
early studies on the detoxification of aflatoxins
was that of Ciegler et al.,20 who showed that the
bacterium Flavobacteriu m aurantiacum removed
aflatoxins from solution. This bacterium was later
shown to actually degrade AFB1 in culture.47
Actively growing yeasts have been shown to de-
grade patulin.10 Among lactobacilli,Z. acidophi-
lus was found to be an efficient inhibitor of
growth and toxin production by A. flavus.31 Colo-
nization of maize by Fusarium spp. has been
shown to be clearly inhibited by Aspergillus and
Penicillium spp. at 25°C, depending on aw and
the species tested.49 Interactions that led to de-
creased colonization by Fusarium did not nega-
tively affect fumonisin production.
Attempts to control the growth of Botrytis ci-
nerea on apples have included testingPseudomo-
nas cepacia, Erwinia sp., Pichia guilliermondii,
Cryptococcus sp., Acremonium breve, and Tri-
choderma pseudokoningii, and all were found to
REFERENCES
1. Alberts, J.F., W.C.A. Gelderblom, P.G.Thiel, et al. 1990.
Effects of temperature and incubation period on pro-
duction of fumonisin B1 by Fusarium moniliforme.
Appl. Environ. Microbiol. 56:1729-1733.
2. Altug, T, A.E. Yousef, and E.H. Marth. 1990. Degra-
dation of aflatoxin B1 in dried figs by sodium bisulfite
with or without heat, ultraviolet energy or hydrogen
peroxide. J. Food Protect. 53:581-582.
3. Ayres, J.C., J.O. Mundt, and W.E. Sandine. 1980. Mi-
crobiology of foods, 658-683. San Francisco: Freeman.
4. Bacon, C.W., J.G. Sweeney, J.D. Hobbins, et al. 1973.
Production of penicillic acid and ochratoxin A on poul-
try feed by Aspergillus ochraceus: Temperature and
moisture requirements. Appl. Microbiol. 26:155-160.
5. Buchanan, R.L., M.A. Harry, and M.A. Gealt. 1983.
Caffeine inhibition of sterigmatocystin, citrinin, and
patulin production. J. Food Sci. 48:1226-1228.
6. Bullerman, L.B. 1976. Examination of Swiss cheese
for incidence of mycotoxin producing molds. J. Food
ScL 41:26-28.
7. Bullerman, L.B. 1984. Effects of potassium sorbate
on growth and patulin production by Penicillium
patulum and Penicillium roquefortii. J. Food Protect.
47:312-316.
8. Bullerman, L.B., P.A. Hartman, and J.C. Ayres. 1969.
Aflatoxin production in meats. I. Stored meats. Appl.
Microbiol. 18:714-717.
9. Bullerman, L.B., PA. Hartman, and J.C. Ayres. 1969.
Aflatoxin production in meats. II. Aged dry salamis
and aged country cured hams. Appl. Microbiol.
18:718-722.
10. Burroughs, L.F. 1977. Stability of patulin to sulfur
dioxide and to yeast fermentation. J. Assoc. Off. Anal.
Chem. 60:100-103.
11. Busby, W.F., Jr., and G.N. Wogan. 1979. Food-borne
mycotoxins and alimentary mycotoxicoses. In Food-
borne infections and intoxications, ed. H. Riemann and
FL. Bryan, 519-610. New York: Academic Press.
12. Butler, W.H. 1974. Aflatoxin. In Mycotoxins, ed. I.F.H.
Purchase, 1-28. New York: Elsevier.
13. Camou-Arriola, J.P., and R.L. Price. 1989. Destruc-
tion of aflatoxin and reduction of mutagenicity of natu-
rally-contaminated corn during production of a corn
snack. J. Food Protect. 52:814-817.
14. Castelo, M.M., S.S. Sumner, and L.B. Bullerman.
1998. Occurrence of fumonisins in corn-based food
products. J. Food Protect. 61:704-707.
be effective.25 The most effective was Erwinia
sp., especially under ambient conditions.
15. Castelo, M.M., S.S. Sumner, and L.B. Bullerman.
1998. Stability of fumonisins in thermally processed
corn products. J. Food Protect. 61:1030-1033.
16. Chelkowski, J., and A. Visconti, eds. 1992. Alterna-
ria: Biology, plant diseases and metabolites. New York:
Elsevier.
17. Christensen, CM. 1971. Mycotoxins. CRC Crit. Rev.
Environ. Cont. 2:57-80.
18. Chulze, S.N., A.M. Torres, A.M. Dalcero, et al. 1995.
Alternaria mycotoxins in sunflower seeds: Incidence
and distribution of the toxins in oil and meal. J. Food
Protect. 58:1133-1135.
19. Ciegler, A., H.-J. Mintzlaff, D. Weisleder, et al. 1972.
Potential production and detoxification of penicillic
acid in mold-fermented sausage (salami). Appl.
Microbiol. 24:114-119.
20. Ciegler, A., E.B. Lillehoj, R.E. Peterson, et al. 1966.
Microbial detoxification of aflatoxin. Appl. Microbiol.
14:934-939.
21. Davis, N.D., and U.L. Diener. 1987. Mycotoxins. In
Food and beverage mycology, 2d ed., ed. L.R. Beuchat,
517-570. New York: Van Nostrand Reinhold.
22. Davis, VM., and M.E. Stack. 1991. Mutagenicity of
stemphyltoxin III, a metabolite of Alternaria alternata.
Appl. Environ. Microbiol. 57:180-182.
23. Deger, G.E. 1976. Aflatoxin—human colon carcino-
genesis? Ann. Intern. Med. 85:204-205.
24. de Nijs, M., H.P. van Egmond, M. Nauta, et al. 1998.
Assessment of human exposure to fumonisin B 1 .
J. Food Protect. 61:879-884.
25. Dock, L.L., PV Nielsen, and J.D. Floros. 1998. Bio-
logical control of Botrytis cinerea growth on apples
stored under modified atmospheres. J. Food Protect.
61:1661-1665.
26. El-Banna, A.A., and P.M. Scott. 1984. Fate of my-
cotoxins during processing of foodstuffs. III. Ochra-
toxin A during cooking of faba beans (Viciafaba) and
polished wheat. J. Food Protect. 47:189-192.
27. Enomoto, M., and M. Saito. 1972. Carcinogens pro-
duced by fungi. Annu. Rev. Microbiol. 26:279-312.
28. Escher, F.E., PE. Koehler, and J.C. Ayres. 1973. Pro-
duction of ochratoxins A and B on country cured ham.
Appl. Microbiol. 26:27-30.
29. Farber, J.M., and G.W. Sanders. 1986. Fusarin C pro-
duction by North American isolates of Fusarium
moniliforme. Appl. Environ. Microbiol. 51:381-384.
30. Gelderblom, W.C.A., N.P.J. Kriek, W.F.O. Marasas, et
al. 1991. Toxicity and carcinogenicity of the Fusarium
moniliforme metabolite, fumonisin B1, in rats. Carcino-
genesis 12:1247-1251.
31. Gelderblom, W.C.A., K. Jaskiewicz, W.F.O. Marasas,
et al. 1988. Fumonisins—novel mycotoxins with can-
cer-promoting activity produced by Fusarium
moniliforme. Appl. Environ. Microbiol. 54:1806-1811.
32. Gourama, H., and L.B. Bullerman. 1995. Antimycotic
and antiaflatoxigenic effect of lactic acid bacteria:
A review. J. Food Protect. 58:1275-1280.
33. Harrison, M.A. 1989. Presence and stability of patu-
Hn in apple products: A review. J. Food Saf 9:
147-153.
34. Hesseltine, CW. 1967. Aflatoxins and other mycotox-
ins. Health Lab. ScL 4:222-228.
35. Holmquist, G.U., H.W. Walker, and H.M. Stahr. 1983.
Influence of temperature, pH, water activity and anti-
fungal agents on growth of Aspergillus flavus and A.
parasiticus. J. Food Sci. 48:778-782.
36. Jorgenssen, K.V, D.L. Park, S.M. Rua, Jr., et al. 1990.
Reduction of mutagenic potentials in milk: Effects of
ammonia treatment on aflatoxin-contaminated cotton-
seed. J. Food Protect. 53:777-778, 817.
37. Karunaratne, A., E. Wezenberg, and L.B. Bullerman.
1990. Inhibition of mold growth and aflatoxin produc-
tion by Lactobacillus spp. J. Food Protect. 53:
230-236.
38. Kedera, C.J., R.D. Plattner, and A.E. Desjardins. 1999.
Incidence of Fusarium spp. and levels of fumonisin
Bi in maize in western Kenya. Appl. Environ.
Microbiol. 65:41-44.
39. Kim, J-C, and Y-W Lee. 1994. Sambutoxin, a new
mycotoxin produced by toxic Fusarium isolates ob-
tained from rotted potato tubers. Appl. Environ.
Microbiol. 60:4380-4386.
40. Kim, J-C, Y-W Lee, and S-H Yu. 1995. Sambutoxin-
producing isolates of Fusarium species and occurrence
of sambutoxin in rotten potato tubers. Appl. Environ.
Microbiol. 61:3750-3751.
41. Kurtzman, C P , B.W Horn, and CW. Hesseltine. 1987.
Aspergillus nominus, a new aflatoxin-producing spe-
cies related to Aspergillus flavus and Aspergillus
tamarii. Antonie van Leeuwenhoek 53:147-158.
42. Labuza, T.P. 1983. Regulation of mycotoxins in food.
J. Food Protect. 46:260-265.
43. Lee, YJ., and WM. Hagler, Jr. 1991. Aflatoxin and
cyclopiazonic acid production by Aspergillus flavus
isolated from contaminated maize. J. Food Sci. 56:
871-872.
44. Leistner, L., and C. Eckardt. 1979. Vorkommen
toxigener Penicillien bei Fleischerzeugnissen.
Fleischwirtsch. 59:1892-1896.
45. Leistner, L., and F. Tauchmann. 1979. Aflatoxinbildung
in Rohwurst durch verschiedene Aspergillus flavus-
Stamme und einer Aspergillus parasiticus-Stamm.
Fleischwirtschaft 50:965-966.
46. Lie, J.L., and E.H. Marth. 1967. Formation of afla-
toxin in cheddar cheese by Aspergillus flavus and
Aspergillus parasiticus. J. Dairy Sci. 50:1708-1710.
47. Line, IE., R.E. Brackett, and R.E. Wilkinson. 1994.
Evidence for degradation of aflatoxin B1 by Flavobac-
terium aurantiacum. J. Food Protect. 57:788-791.
48. Marasas, W.F.O., K. Jaskiewicz, FS. Venter, et al. 1988.
Fusarium moniliforme contamination of maize in oe-
sophageal cancer areas in Transkei. S. Afr. Med. J.
74:110-114.
49. Marin, S., V Sanchis, F. RuIl, et al. 1998. Coloniza-
tion of maize grain by Fusarium moniliforme and
Fusarium proliferatum in the presence of competing
fungi and their impact on fumonisin production.
J. Food Protect. 61:1489-1496.
50. Marin, S., V Sanchis, I. Vines, et al. 1995. Effect of
water activity and temperature on growth and
fumonisin B 1 and B 2 production by Fusarium
proliferatum and F. moniliforme on maize grain. Lett.
Appl. Microbiol. 21:298-301.
51. Marth, E.H., and B.G. Calanog. 1976. Toxigenic fungi.
In Food microbiology: Public health and spoilage as-
pects, ed. M.P deFigueiredo and D.F. Splittstoesser,
210-256. Westport, CT: AVI.
52. Nelson, PE., R.D. Plattner, D.D. Shackelford, et al.
1992. Fumonisin B1 production by Fusarium species
other than F moniliforme in section Liseola and by
some related species. Appl. Environ. Microbiol.
58:984-989.
53. Newsome, R. 1999. Issues in international trade: Look-
ing to the Codex Alimentarius Commission. Food
Technol. 53, no. 6:26.
54. Niranjan, B.G., N.K. Bhat, and N.G. Avadhani. 1982.
Preferential attack of mitochondrial DNA by aflatoxin
B1 during hepatocarcinogenesis. Science 215:73-75.
55. Northolt, M.D., C.A.H. Verhulsdonk, PS.S. Soentoro,
et al. 1976. Effect of water activity and temperature
on aflatoxin production by Aspergillus parasiticus. J.
Milk Food Technol. 39:170-174.
56. Pestka, JJ., M.N. Abouzied, and Sutikno [sic]. 1995.
Immunological assays for mycotoxin detection. Food
Technol. 49, no. 2:120-128.
57. Pestka, J.J., J.I. Azcona-Olivera, R.D. Plattner, et al.
1994. Comparative assessment of fumonisin in grain-
based foods by ELISA, GC-MS, and HPLC J. Food
Protect. 57:169-172.
58. Pittet, A., V Parisod, and M. Schellenberg. 1992. Oc-
currence of fumonisins B1 and B2 in corn-based prod-
ucts from the Swiss market. J. Agric. Food Chem.
40:1352-1354.
59. Park, K.Y., and L.B. Bullerman. 1983. Effect of cy-
cling temperatures on aflatoxin production by Aspergil-
lus parasiticus and Aspergillus flavus in rice and ched-
dar cheese. J. Food ScL 48:889-896.
60. Ram, B.P., P. Hart, RJ. Cole, et al. 1986. Application
of ELISA to retail survey of aflatoxin Bi in peanut
butter. J. Food Protect. 49:792-795.
61. Rheeder, J.P., W.F.O. Marasas, PG. Thiel, et al. 1992.
Fusarium moniliforme and fumonisins in corn in rela-
tion to human esophageal cancer in Transkei.
Phytophathol. 82:353-357.
62. Roland, J.O., and L.R. Beuchat. 1984. Biomass and
patulin production by Byssochlamys nivea in apple
juice as affected by sorbate, benzoate, SO2 and tem-
perature. J. Food ScL 49:402-406.
63. Ross, P.F., L.G. Rice, R.D. Plattner, et al. 1991. Con-
centrations of fumonisin B1 in feeds associated with
animal health problems. Mycopathol. 114:129-135.
64. Ross, PE, PE. Nelson, J.L. Richard, et al. 1990. Pro-
duction of fumonisins by Fusarium moniliforme and
Fusarium proliferatum isolates associated with equine
leukoencephalomalacia and a pulmonary edema syn-
drome in swine. Appl. Environ. Microbiol. 56:
3225-3226.
65. Schillinger, U, R. Geisen, and W.H. Holzapfel. 1996.
Potential of antagonistic microorganisms and bacte-
riocins for the biological preservation of foods. Trends
Food Sci. Technol 7:158-164.
66. Schindler, A.E 1977. Temperature limits for produc-
tion of aflatoxin by twenty-five isolates of Aspergillus
flavus and Aspergillus parasiticus. J. Food. Protect.
40:39-40.
67. Serck-Hanssen. A. 1970. Aflatoxin-induced fatal
hepatitis? Arch. Environ. Health 20:729-731.
68. Shelef, L.A., and B. Chin. 1980. Effect of phenolic
antioxidants on the mutagenicity of aflatoxin B1. Appl.
Environ. Microbiol. 40:1039-1043.
69. Shih, C.N., and E.H. Marth. 1974. Some cultural con-
ditions that control biosynthesis of lipid and aflatoxin
by Aspergillus parasiticus. Appl. Microbiol. 27:
452-456.
70. Shotwell, O.L., and CW. Hesseltine. 1983. Five-year
study of mycotoxins in Virginia wheat and dent corn.
J.Assoc. Off. Anal. Chem. 66:1466-1469.
71. Siame, B.A., S.E Mpuchane, B.A. Gashe, et al. 1998.
Occurrence of aflatoxins, fumonisin Bi, and
zearalenone in foods and feeds in Botswana. J. Food
Protect. 61:1670-1673.
72. Sommer, N.F., J.R. Buchanan, and RJ. Fortlage. 1974.
Production of patulin by Penicillium expansum. Appl.
Microbiol. 28:589-593.
73. Stahr, H.M., R.L. Pfeiffer, PJ. Imerman, et al. 1990.
Aflatoxins—the 1988 outbreak. Dairy Food Environ.
Sanit. 10:15-17.
74. Stark, A.A. 1980. Mutagenicity and carcinogenicity
of mycotoxins: DNA binding as a possible mode of
action. Annu. Rev. Microbiol. 34:235-262.
75. Stinson, E.E., D.D. Bills, S.F. Osman, et al. 1980.
Mycotoxin production by Alternaria species grown on
apples, tomatoes, and blueberries. J. Agric. Food Chem.
28:960-963.
76. Stinson, E.E., S.F. Osman, E.G. Beisler, et al. 1981.
Mycotoxin production in whole tomatoes, apples, or-
anges, and lemons. J. Agric. Food Chem. 29:790-792.
77. Stoloff, L. 1987. Carcinogenicity of aflatoxins.
Science 237:1283-1284 (letter to editor with two
responses).
78. Sydenham, E.W., W.F.O. Marasas, G.S. Shephard, et
al. 1992. Fumonisin concentrations in Brazalian feeds
associated with field outbreaks of confirmed and sus-
pected animal mycotoxicoses. J. Agric. Food Chem.
40:994-997.
79. Sydenham, E.W, G.S. Shepard, PG. Thiel, et al. 1991.
Fumonisin contamination of commercial corn-based
human foodstuffs. J. Agric. Food Chem. 39:2014-2018.
80. Sydenham, E.W., PG. Thiel, W.F.O. Marasas, et al.
1990. Natural occurrence of some Fusarium mycotox-
ins in corn from low and high esophageal cancer preva-
lence areas of the Transkei, southern Africa. J. Agric.
Food Chem. 38:1900-1903.
81. Thiel, PG., W.F.O. Marasas, E.W. Sydenham, et al.
1992. The implications of naturally occurring levels
of fumonisins in corn for human and animal health.
Mycopathologia 117:3-9.
82. Thompson, D.P. 1997. Effect of phenolic compounds
on mycelial growth of Fusarium and Penicillium spe-
cies. J. Food Protect. 60:1262-1264.
83. Tong, C-H., and EA. Draughon. 1985. Inhibition by
antimicrobial food additives of ochratoxin A produc-
tion by Aspergillus sulphureus and Penicillium
viridicatum. Appl. Environ. Microbiol. 49:1407-1411.
84. Torres, M.R., V Sanchis, and AJ. Ramos. 1998. Oc-
currence of fumonisins in Spanish beers analyzed by
an enzyme-linked immunosorbent assay method. Int.
J. Food Microbiol. 39:139-143.
85. Trenk, H.L., and PA. Hartman. 1970. Effects of mois-
ture content and temperature on aflatoxin production
in corn. Appl. Microbiol. 19:781-784.
86. Wheeler, K.A., B.F. Hurdman, and J.L Pitt. 1991. In-
fluence of pH on the growth of some toxigenic spe-
cies of Aspergillus, Penicillium and Fusarium. Int. J.
Food Microbiol. 12:141-150.
87. Wiebe, L.A., and L.F. Bjeldanes. 1981. Fusarin C, a
mutagen from Fusarium moniliforme grown on corn.
J. Food ScL 46:1424-1426.
88. Wheeler, J.L., M.A. Harrison, and PE. Koehler. 1987.
Presence and stability of patulin in pasteurized apple
cider. J. Food ScL 52:479-480.
89. Wogan, G.N. 1966. Chemical nature and biological
effects of the aflatoxins. Bacteriol. Rev. 30:460-470.
90. Wu, M.T., XC. Ayres, and RE. Koehler. 1974. Produc-
tion of citrinin by Penicillium viridicatum on country-
cured ham. Appl. Microbiol. 27:427-428.
91. Yoshizawa, T., A. Yamashita, and Y. Luo. 1994.
Fumonisin occurrence in corn from high- and low-risk
areas for human esophageal cancer in China. Appl.
Environ. Microbiol 60:1626-1629.