1054

Journal of Food Protection, Vol. 68, No. 5, 2005, Pages 1054–1059

Copyright Q, International Association for Food Protection

Effect of Water Activity and Temperature on Growth and the

Relationship between Fumonisin Production and the Radial
Growth of Fusarium verticillioides and
Fusarium proliferatum on Corn
SIMBARASHE SAMAPUNDO,1 FRANK DEVLIEHGERE,1* BRUNO DE MEULENAER,2 AND JOHAN DEBEVERE1

1Laboratory of Food Microbiology and Food Preservation and 2Laboratory of Food Chemistry, Department of Food Safety and Food Quality, Faculty
of Bioscience Engineering, Ghent University, Coupure Links 653, B 9000 Gent, Belgium

MS 04-436: Received 14 September 2004/Accepted 22 January 2005

ABSTRACT
The two major fumonisin-producing Fusarium species are Fusarium verticillioides and Fusarium proliferatum. The growth
and fumonisin production of these two isolates on corn was studied at water activities (aw) between 0.860 and 0.975 and at
temperatures between 15 and 308C. Growth rates (g, mm/day) were obtained by linear regression during the linear phase of
growth. In general, growth rates for both isolates increased significantly (P , 0.05) with increases in aw and temperature.
Both fumonisin production and radial growth (mycelial development) for both isolates increased with aw at all temperatures
investigated, but the effect of temperature on this relationship was not obvious. The effect of temperature on fumonisin
production at high aw values optimal for growth was only marginal, whereas at lower aw values the effect of temperature was
more pronounced, with more fumonisin production occurring at temperatures not optimal for growth. The optimum temperature
for fumonisin production was between 15 and 258C. For F. proliferatum, the optimum temperature for growth at all aw values,
308C, resulted in the poorest fumonisin production. For both isolates, the slowest initial rate of fumonisin production was at
158C, the temperature at which the slowest growth rates were obtained.

Although the Fusarium species are best known for the
production of the tricothecene mycotoxins (26), species
such as Fusarium verticillioides and Fusarium proliferatum
produce a series of other important secondary metabolites
such as moniliformin, fusarin C, fusaric acid, and fumoni-
sins (3, 5, 6, 16, 18, 32). Fumonisins are important because
of their links to both human and animal toxicoses from
consumption of contaminated corn-based food and feeds
(28, 29).
Corn intended for human consumption and containing
high levels of fumonisins has been associated with high
incidences of esophageal cancer in the Transkei region of
South Africa (19, 29–31), in Northern Italy (9), in the Linx-
hian region of China (8, 29), and in the southeastern United
States (11). The incidence of neural tube defects in Transkei
South Africa also is very high (34). The association of fu-
monisins with animal toxicoses such as equine leucoence-
phalomalacia (28) and porcine pulmonary edema (12, 28)
has been well documented. Fumonisins also have been re-
ported to be hepatotoxic, nephrotoxic, and carcinogenic to
rats (10, 11, 14).
The effects of environmental factors, competition, and
antifungal agents on growth and/or fumonisin production
by F. verticillioides and F. proliferatum have been previ-
ously investigated (2, 4, 7, 20–24). In experiments used to
investigate the relationship between growth or time and tox-
* Author for correspondence. Tel: 11 32-9-264-6178; Fax: 11 32-9-
225-5510; E-mail: frank.devlieghere@ugent.be.
in production, effects have been evaluated after relatively
long periods of time; in most cases, single measurements
were made after two or more weeks. The major objective
of the present study was to obtain a better understanding
of the relationship between growth and fumonisin produc-
tion as influenced by water activity (aw) and temperature
by evaluating both growth and fumonisin production over
shorter time and growth intervals.
MATERIALS AND METHODS
Experimental design. A full factorial design was used to
investigate F. verticillioides and F. proliferatum growth and fu-
monisin production on corn. Four temperatures (15, 22, 25, and
308C) and four aw values between 0.860 and 0.975 at which
growth took place were examined with 20 replicates prepared per
condition. Samples for fumonisin analysis were collected six times
during the growth to coincide either with time intervals of 1 week
for colonies that grew slowly or with colony diameters of ap-
proximately 15, 30, 45, 60, 75, and 85 mm for colonies that grew
rapidly.
Corn grain. Dried yellow corn was obtained from Aveve
NV (Leuven, Belgium). The corn had an initial moisture content
of 12.79 6 0.45% dry basis and an aw of 0.698 6 0.015. Both
adsorption and desorption isotherms were developed to fully char-
acterize the water content of the corn substrate.
Fungal isolates. F. verticillioides Sheldon (25N) and F. pro-
liferatum (Matsushima) Nirenberg (73N) were isolated from corn
and are proven high fumonisin producers. The two isolates were
obtained from the Food Technology Department Culture Collec-
J. Food Prot., Vol. 68, No. 5 FUSARIUM SPP. GROWTH AND FUMONISIN PRODUCTION
FIGURE 1. Plots of colony growth rate g (mm/day) versus temperature (8C) for (a) F. verticillioides and (b) F. proliferatum at different
aw values.
tion of the University of Lleida (Lleida, Spain). Both isolates were
maintained on potato dextrose agar (PDA) (Oxoid, Basingstoke,
UK).
Preparation of corn substrate. The corn was sterilized us-
ing 25 kGy of g-irradiation at IBA Mediris (Fleurus, Belgium) to
ensure no fungal infection or contamination of the substrate while
retaining the ability of the corn to germinate. Corn kernels were
then stored aseptically at 4 to 78C until further use. The adjust-
ment of aw to the desired value was achieved by the direct addition
of a certain amount of sterile distilled water as determined by the
adsorption isotherms developed for corn. The corn kernels were
then allowed to equilibrate in two phases: they were kept at 48C
for 2 days with periodic mixing and then placed in perforated
aluminum cups over a glycerol-water solution with aw similar to
that desired in the substrate and incubated for 7 days at the final
incubation temperature. The final aw was determined using the
Novasina Thermoconstanter (TH200, Axair Ltd. Systems, Pfaffi-
kon, Switzerland).
Preparation of inoculum, inoculation, incubation, and
growth assessment. An inoculation loop was used to aseptically
scrape sporulating mycelia from the surface of PDA slants on
which the isolates were maintained. The loop was then used to
centrally inoculate the surfaces of petri plates (90 mm) containing
PDA. The plate were incubated at 308C for 6 days and then at
15, 22, 25, or 308C for 1 day to enable the isolates to adapt to
the incubation conditions to be investigated. About 24 g of re-
hydrated corn was placed in each sterile plate to form a single
layer of kernels. A 5.6-mm-diameter agar disk cut from the margin
of a 7-day-old colony growing on PDA was transferred to the
center of each plate using a sterile cork borer. Plates containing
kernels of the same aw were then placed over glycerol-water so-
lutions of the same aw in sealed containers, with 20 plates in each
container. Containers were then incubated at 15, 22, 25, or 308C.
From each container, four randomly chosen plates containing
growing colonies were assessed periodically for changes in
growth. The growth was assessed as the change in diameter of the
growing colony and was determined by measuring two perpen-
dicular diameters per chosen plate. The average of the four chosen
colonies was used to evaluate the change in colony diameter at a
particular time.
1055
Fumonisin analyses. The total fumonisin concentration in
the samples was determined in duplicate for each sample using a
commercial enzyme-linked immunosorbent assay (ELISA) kit
(RIDASCREEN Fumonisin Fast, BioPharm, Darmstadt, Germa-
ny) containing a competitive immunoassay for the quantitative
analysis of fumonisin residues in corn and corn products. For this
ELISA, the mean lower detection limit for fumonisins is 0.2 mg/
g, and the upper quantification limit is 9 mg/g. Sample extraction
and preparation and assay conditions were as suggested by the
manufacturer. However, samples with fumonisin concentrations
higher than 9 mg/g were diluted until the concentration fell within
the limits of the assay.
Mathematical and statistical methods. The slopes of plots
of colony diameter during the linear growth phase versus time
were estimated by simple linear regression using Excel 2000 (Mi-
crosoft, Redmond, Wash.), and the colony growth rate (g, mm/
day) was determined. Division of g by 2 gives the radial growth
rate (mm/day). Analysis of variance (ANOVA) was used to de-
termine whether the environmental factors had a significant effect
on colony growth rates.
RESULTS AND DISCUSSION
Effect of aw and temperature on growth rates of F.
verticillioides and F. proliferatum. The growth curves
based on colony diameters were typical of fungal growth
for both isolates and were characterized by a lag phase for
those colonies where growth was hindered by low aw and/
or temperature and followed by linear growth for all colo-
nies. Figure 1 shows the estimated colony growth rate g at
different aw values as a function of the temperature for F.
verticillioides and F. proliferatum. The upper asymptote of
growth was not achieved in most cases because of the lim-
ited radial growth surface of the 90-mm-diameter plates or
the maximum period of 6 weeks of incubation with no sub-
strate limitation. At high aw values and temperatures suit-
able for growth, the colonies grew rapidly, reaching the
plate diameter after a few days, whereas at less optimal
conditions the colonies grew at a very slow rate and some
did not reach 90 mm within 6 weeks.
1056 SAMAPUNDO ET AL. J. Food Prot., Vol. 68, No. 5

TABLE 1. Analysis of variance of growth rates of Fusarium ver- et al. (15), Cahagnier et al. (7), Marin et al. (20, 22, 24),
ticillioides and F. proliferatum at different aw and temperature and Velluti et al. (33).
valuesa
Effect of aw on the relationship between fumonisin
Variable df MSb MSc Fb Fc
production and radial growth. The effect of aw on the
Temperature 3 31.9 13.6 27.8d 28.0d relationship between colony diameter or time and fumoni-
aw 3 12.2 3.1 10.7d 6.5d sin production at different temperatures is shown for F.
Residual 9 1.1 0.5 proliferatum and F. verticillioides in Figures 2 and 3, re-
Total 15 spectively. For both isolates at the temperatures investigat-
a
ed, fumonisin production was highly aw dependent. Gen-
df, degrees of freedom; MS, mean square. erally, more fumonisin was produced for the same amount
b F. verticillioides.
c F. proliferatum. of growth (colony diameter) or time elapsed under condi-
d P , 0.05. tion of higher aw values. When F. verticillioides colonies
grown at 308C reached 80 mm diameter, fumonisin at
168.1, 3.7, 0.9, and 1.0 mg/g were produced at aw values
Table 1 shows the results of the ANOVA for the sig- of 0.969, 0.949, 0.937, and 0.922, respectively. The greatest
nificance of the effects of temperature and/or aw on the differences occurred between aw values of 0.969 and 0.949,
growth rates of F. verticillioides and F. proliferatum. Both where the decrease in fumonisin production ranged from 4-
aw and temperature had a significant effect (P , 0.05) on to almost 50-fold. The only exception to this trend was
the growth rates of both isolates. Despite the significant noted for F. proliferatum at 15 and 258C, where the amount
effect of each variable, there was no interaction between of fumonisin produced for the same amount of growth was
the variables for either isolate. However, Marin et al. (20, initially greater at aw 0.948 than at 0.972. However more
22, 24), and Velluti et al. (33) found interactions between fumonisin was produced for the same amount of growth at
aw and temperature in relation to growth rates of F. proli- aw 0.972 than at 0.948 after a colony diameter of approx-
feratum and F. verticillioides. Although the reason for these imately 70 mm had been reached at both temperatures.
different results is not clear, the range of conditions in the The general trend of a marked decrease in the amount
present study may not have been large enough for inter- of fumonisin produced and in the rate of production in re-
actions to be observed. sponse to a decrease in aw at the temperatures investigated
The optimum temperature for growth for both isolates has been reported by others but with larger colonies and
was 308C at all aw values tested. Using in vitro studies, over longer periods of time. Marin et al. (20) observed the
other researchers have found various optimum temperatures same trend for one F. verticillioides isolate (25N) and two
for growth. Marin et al. (20, 22) reported an optimum of F. proliferatum isolates (75N and 131N), which all gener-
308C on various growth media, including autoclaved corn, ally produced more fumonisin at aw 0.98 than at 0.95 and
for both F. proliferatum and F. verticillioides. Alberts et al. 0.92 after 4 weeks at 15 to 308C. Fumonisin B1 production
(1) reported optimum growth at both 20 and 258C for F. increased with aw at temperatures between 7 and 308C for
verticillioides on autoclaved corn, and Le Bars et al. (15) isolates of F. verticillioides and F. proliferatum grown on
reported optimum growth at 258C for F. verticillioides on corn (21). Marin et al. (20) also reported that F. prolifer-
autoclaved corn. Generally, an increase in aw or temperature atum grown at 158C produced more fumonisin at aw 0.95
results in an increase in the growth rate for both isolates. and 0.92 than at 0.98 after 2 and 4 weeks, results similar
This results confirms similar findings reported by Le Bars to ours. Thus, aw stress may be stimulating fumonisin pro-

FIGURE 2. Plots of (a) colony diameter (mm) and (b) time (days) versus fumonisin (mg/g) produced by F. proliferatum at different
temperatures and aw values.
J. Food Prot., Vol. 68, No. 5 FUSARIUM SPP. GROWTH AND FUMONISIN PRODUCTION
FIGURE 3. Plots of colony diameter (mm) (a and b) and time (days) (c and d) versus fumonisin (mg/g) produced by F. verticillioides
at different temperatures and aw values.
duction by F. proliferatum at temperatures suboptimal for
growth during the initial growth phases. There also may be
a species-specific component, because this result was not
observed for F. verticillioides.
Effect of temperature on the relationship between
growth and fumonisin production. The effect of temper-
ature on fumonisin production as a function of growth or
time at different aw values is shown for F. proliferatum and
F. verticillioides in Figures 2 and 3, respectively. Although
the effect of temperature on the rate of mycelial develop-
ment follows a very clear pattern, its effect on fumonisin
production is not as clear. For both isolates, the effect of
temperature on the rate of fumonisin production, the
amount of fumonisin produced for the same amount of
growth, and the final amount of fumonisin produced varied
markedly with aw.
Figure 2a shows the fumonisin production results for
F. proliferatum based on colony diameter. For this isolate
at aw 0.972, the most fumonisin was produced at 228C, and
production decreased in order at 15, 25, and 308C for the
same amount of growth. At aw 0.972, 192.6, 160.7, 153.9,
1057
and 144.1 mg/g were produced after approximately 80 mm
of colony growth at 22, 25, 15, and 308C, respectively. At
aw 0.948, the most fumonisin was produced at 158C, and
production decreased in order at 25, 22, and 308C for the
same amount of growth. At aw 0.928, much less fumonisin
was produced in comparison to that produced at higher aw
values; 32.4, 10.9, 6.7, and 0.8 mg/g were produced after
approximately 80 mm of colony growth at 25, 22, 15, and
308C, respectively.
Figure 2b shows the fumonisin production results for
F. proliferatum as a function of time. For this isolate at aw
0.972, the production rates were fastest at 22, 25, and 308C
and slowest at 158C. The time for significant fumonisin
production to occur was shorter, approximately 7 to 10
days, at the higher temperatures and longer at 158C (ap-
proximately 14 days). At aw 0.948, the fumonisin produc-
tion rate was highest at 258C and did not differ significantly
at 22, 30, and 158C. However, it generally took longer for
significant fumonisin production to occur at 30 and 158C
than at 22 and 258C.
Although 308C was the optimum temperature for
1058 SAMAPUNDO ET AL.
growth of F. proliferatum, it resulted in production of the
smallest amount of fumonisin at any aw. The smallest
amounts of fumonisin at any colony diameter and after ap-
proximately 80 mm of colony growth or 6 weeks of incu-
bation at any aw were also produced at 308C. The effect of
temperature on fumonisin production appears to be margin-
al at higher aw; fumonisin production by F. proliferatum at
aw 0.969 and 308C was still high. The total fumonisin pro-
duced and rate of production were lowest at 158C.
The findings from the few available studies of F. pro-
liferatum are consistent with those of the present study. Ma-
rin et al. (21) reported that the optimum temperature for
fumonisin B1 production was 158C followed by 20, 25, 10,
and 308C at aw 0.97, which is consistent with our finding
that the smallest amount of fumonisin was produced at
308C. Marin et al. (20) also found an increase in fumonisin
produced by F. proliferatum (131N) after 4 weeks as tem-
perature was increased from 15 to 308C at aw 0.98. How-
ever, in accordance with our results, these authors noted
that more fumonisin was produced after 4 weeks at 15 and
258C than at 308C at lower aw values of 0.95 and 0.92.
As observed for F. proliferatum, temperature had a
marked effect on the initial fumonisin production rate for
F. verticillioides (Fig. 3c and 3d). The slowest production
rate irrespective of aw was at 158C, at which the slowest
growth rates also were observed. The smallest amount of
fumonisin after approximately 80 mm of colony growth
was produced at 22, 25, 30, and 158C at aw values of 0.969,
0.949, 0.937, and 0.922, respectively (Fig. 3a and 3b). The
optimum temperature for fumonisin production varied with
aw: 308C at aw 0.969, 158C at aw 0.949, and 228C at aw
0.937 and 0.922. Results from other studies are also am-
biguous but in most cases are consistent with our findings.
Le Bars et al. (15) reported that for F. verticillioides the
optimum fumonisin B1 production rate occurred at 208C
and then decreased sharply in order at 25, 15, 30, and 108C.
Marin et al. (20) observed an increase in amount of fu-
monisin produced after 4 weeks as temperature increased
from 15 to 308C at aw 0.98 for F. verticillioides (25N).
However, they also noted that more fumonisin was pro-
duced after 4 weeks at 15 and 258C than at 308C at lower
aw values of 0.95 and 0.92. Marin et al. (21) also reported
that the optimum temperature for fumonisin B1 production
by F. verticillioides at aw 0.97 varied between 20 and 308C.
Alberts at al. (1) reported that the maximal production of
fumonisin B1 by F. verticillioides on corn cultures occurred
at 208C, with very low production at 308C; however, they
did not have accurate details of the water content or aw.
Le Bars et al. (15) suggested that F. verticillioides de-
graded the fumonisin it produced after a period of time,
because toxin amounts were stable in corn. Alberts et al.
(1) reported that this degradation started after 13 weeks of
incubation at 20 to 258C, whereas Marin et al. (22) reported
that degradation began after approximately 5 weeks. In the
present study, only a few plates were incubated for the max-
imum period of 6 weeks; the incubation time for most of
the conditions optimal for growth was less than 5 weeks
and in some cases only days. The only conditions under
which fumonisin degradation was noted were F. prolifer-
J. Food Prot., Vol. 68, No. 5
atum at aw 0.928 and 308C, at which a maximum fumonisin
concentration of 3.2 mg/g was attained after 14 days. There-
after, the concentration progressively decreased to 0.8 mg/
g after 38 days.
The stimulation of mycotoxin formation under condi-
tions of growth stress has long been postulated. However,
the results of this and other studies indicate that different
stresses have different effects; aw is positively correlated
with fumonisin production and production rate and tem-
perature has a less direct effect. At aw values suboptimal
for growth, fumonisin production is higher at intermediate
temperatures.
ACKNOWLEDGMENTS
The authors are grateful to the Belgian Technical Cooperation (Brus-
sels, Belgium) for their financial assistance and the Department of Food
Technology Culture Collection, University of Lleida (Lleida, Spain) for
supplying the fungal isolates.
REFERENCES
1. Alberts, J. F., W. C. A. Gelderblom, P. G. Thiel, W. F. O. Marasas,
D. J. Van Schalkwyk, and Y. Vehrend. 1990. Effects of temperature
and incubation period on the production of fumonisin B1 by Fusar-
ium moniliforme. Appl. Environ. Microbiol. 56:1729–1733.
2. Avantaggiato, G., F. Quaranta, E. Desiderio, and A. Visconti. 2002.
Fumonisin contamination of maize hybrids visibly damaged by Se-
samia. J. Sci. Food Agric. 83:13–18.
3. Bacon, C. W., and P. E. Nelson. 1994. Fumonisin production in corn
by toxigenic strains of Fusarium moniliforme and Fusarium proli-
feratum. J. Food Prot. 57:514–521.
4. Bacon, C. W., and J. W. Williamson. 1992. Interactions of Fusarium
moniliforme, its metabolites and bacteria with corn. Mycopathologia
117:65–71.
5. Booth, C. 1971. The genus Fusarium. Commonwealth Mycological
Institute, Kew, UK.
6. Bullerman, L. B., and W.-Y. J. Tsai. 1994. Incidence and levels of
Fusarium moniliforme, Fusarium proliferatum and fumonisins in
corn and corn-based foods and feeds. J. Food Prot. 57:541–546.
7. Cahagnier, B., B. Melcion, and D. Richard-Molard. 1995. Growth
of Fusarium moniliforme and its biosynthesis of fumonisin B1 on
maize grain as a function of different water activities. Lett. Appl.
Microbiol. 20:247–251.
8. Chu, F. S., and G. O. Li. 1994. Simultaneous occurrence of fumon-
isin B1 and other mycotoxins in mouldy corn collected from the
People’s Republic of China in regions with high incidence of oe-
sophageal cancer. Appl. Environ. Microbiol. 60:847–852.
9. Fransceschi, S., E. Bidoldi, A. E. Baron, and C. La Vecchia. 1990.
Maize and risk of cancers of the oral cavity, pharynx, and oesoph-
agus in northeastern Italy. J. Natl. Cancer Inst. 82:1407–1411.
10. Gelderblom, W. C. A., K. Jaskiewicz, W. F. O. Marasas, P. G. Thiel,
R. M. Horak, R. Vleggaar, and N. P. J. Kriek. 1988. Fumonisins—
novel mycotoxins with cancer promoting activity produced by Fu-
sarium moniliforme. Appl. Environ. Microbiol. 54:1806–1811.
11. Gelderblom, W. C. A., W. F. O. Marasas, R. Vleggaar, P. G. Thiel,
and M. E. Cawood. 1992. Fumonisins: isolation, chemical charac-
terization and biological effects. Mycopathologia 117:11–16.
12. Haschek, W. M., G. Motelin, D. K. Ness, K. S. Harlin, W. F. Hall,
R. F. Vesonder, R. E. Peterson, and V. R. Beasley. 1992. Character-
ization of fumonisin toxicity in orally and intravenously dosed
swine. Mycopathologia 117:83–96.
13. Headrick, J. M., J. K. Pataky, and J. A. Juvik. 1990. Relationships
among carbohydrate content of kernels, condition of silks after pol-
lination, and response of sweet corn inbred lines to infection of ker-
nels by Fusarium moniliforme. Phytopathology 80:487–494.
14. International Agency for Research on Cancer. 2002. Some traditional
herbal medicines, some myotoxins, naphthalene and styrene. IARC
Monogr. 82:301.
J. Food Prot., Vol. 68, No. 5 FUSARIUM SPP. GROWTH AND FUMONISIN PRODUCTION
15. Le Bars, J., P. Le Bars, J. Dupuy, H. Boudra, and R. Casini. 1994.
Biotic and abiotic factors in fumonisin B1 production and stability.
J. AOAC Int. 77:517–521.
16. Leslie, J. F., R. D. Plattner, A. E. Desjardins, and C. J. R. Klittich.
1992. Fumonisin B1 production by strains from different mating
populations of Giberrella fujikoroi (Fusarium section Liseola). Phy-
topathology 82:341–345.
17. Marasas, W. F. O. 1993. Occurrence of Fusarium moniliforme and
fumonisins in maize in relation to human health. S. Afr. Med. J. 83:
383–384.
18. Marasas, W. F. O., J. D. Miller, R. T. Riley, and A. Visconti. 2000.
Environmental health criteria 219: fumonisin B1. International pro-
gramme on chemical safety, p. 150. World Health Organization, Ge-
neva.
19. Marasas, W. F. O., F. C. Wehner, S. J. Van Rensburg, and D. J. Van
Schalkwyk. 1981. Mycoflora of corn produced in human oesopha-
geal cancer areas in Transkei South Africa. Phytopathology 71:792–
796.
20. Marin, S., V. Homedes, V. Sanchis, A. J. Ramos, and N. Magan.
1999. Impact of Fusarium moniliforme and F. proliferatum coloni-
sation of maize on calorific losses and fumonisin production under
different environmental conditions. J. Stored Prod. Res. 35:15–26.
21. Marin, S., N. Magan, A. Belli, A. J. Ramos, R. Canela, and V. San-
chis. 1999. Two-dimensional profiles of fumonisin B1 production by
Fusarium moniliforme and F. proliferatum in relation to environ-
mental factors and potential for modelling toxin formation in maize
grain. Int. J. Food Microbiol. 51:159–167.
22. Marin, S., N. Magan, J. Serra, A. J. Ramos, R. Canela, and V. San-
chis. 1999. Fumonisin B1 production and growth of Fusarium mon-
iliforme and Fusarium proliferatum on maize, wheat, and barley
grain. J. Food Sci. 64:921–924.
23. Marin, S., V. Sanchis, A. Teixido, R. Saenz, A. J. Ramos, I. Vinas,
and N. Magan. 1996. Water and temperature relations and micro-
conidial germination of Fusarium moniliforme and F. proliferatum
from maize. Can. J. Food Microbiol. 42:1045–1050.
24. Marin, S., V. Sanchis, A. Teixido, R. Saenz, A. J. Ramos, I. Vinas,
and N. Magan. 1998. Colonisation of maize grain by F. moniliforme
and F. proliferatum in the presence of competing fungi and their
impact on fumonisin production. J. Food Prot. 61:1489–1496.
1059
25. Natori, S., K. Hashimota, and Y. Ueno. 1989. Mycotoxins and phy-
cotoxins ’88. Bioact. Mol. 10:185–189.
26. Nelson, P. E., A. E. Desjardins, and R. D. Plattner. 1993. Fumonisins,
mycotoxins produced by Fusarium species: biology, chemistry, and
significance. Annu. Rev. Phytopathol. 31:233–252.
27. Ono, E. Y. S., E. Y. Sasaki, E. H. Hashimota, L. N. Hara, B. Corrêa,
E. N. Itano, Y. Sigiura, Y. Ueno, and E. Y. Hirooka. 2002. Post-
harvest storage of corn: effects of beginning moisture content on
mycoflora and fumonisin contamination. Food Add. Cont. 19:1081–
1090.
28. Ross, P. F., L. G. Rice, G. D. Osweiler, P. E. Nelson, J. L. Richard,
T. M. Wilson, and R. T. Riley. 1992. A review and update of animal
toxicoses associated with fumonisin-contaminated feeds and produc-
tion of fumonisins by Fusarium isolates. Mycopathologia 117:109–
114.
29. Sydenham, E. W., W. C. A. Gelderblom, P. G. Thiel, and W. F. O.
Marasas. 1990. Evidence for the natural occurrence of fumonisin B1,
a mycotoxin produced by Fusarium moniliforme in corn. J. Agric.
Food Chem. 38:258–290.
30. Sydenham, E. W., W. C. A. Gelderblom, P. G. Thiel, W. F. O. Mar-
asas, and S. Stockenstroms. 1991. Fumonisin contamination of com-
mercial based human foodstuffs. J. Agric. Food Chem. 39:2014–
2018.
31. Thiel, P. G., W. F. O. Marasas, E. W. Sydenham, G. S. Chef-Arts,
and W. C. A. Gelderblom. 1992. Implications of naturally occurring
levels of fumonisins in corn for human and animal health. Myco-
pathologia 117:3–9.
32. U.S. National Toxicology Program. 1999. Toxicology and carcino-
genesis studies of fumonisin B1 (CAS no. 116355-83-0) in F344/N
rats and B6C3F1 mice (feed studies). Technical report no. 496. Na-
tional Institutes of Health publication no. 99-3955. U.S. Department
of Health, Education, and Welfare, Research Triangle Park, N.C.
Available at: http://ntp.niehs.nih.gov/ntp/htdocs/LTprpts/tr496.pdf.
33. Velluti, A., S. Marin, L. Bettuci, A. J. Ramos, and V. Sanchis. 2000.
The effect of fungal competition on colonization of maize grain by
F. moniliforme, F. proliferatum and F. graminearum on fumonisin
B1 and zearalenone formation. Int. J. Food Microbiol. 59:59–66.
34. Venter, P. A., A. L. Christianson, C. M. Hutamo, M. P. Makhura,
and G. S. Gericke. 1995. Congenital anomalies in rural black South
African neonates—a silent epidemic? S. Afr. Med. J. 85:15–20.