Trop Sci 43.

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Trop. Sci. 2003, 43, 76–79

Fusarium spp. and fumonisin B1 in stored maize

from Ogun State, Nigeria
SA Bankole1, OO Mabekoje1 and OA Enikuomehin2
1Department of Biological Sciences, Olabisi Onabanjo University, PMB 2002, Ago-Iwoye, Ogun State, Nigeria.
2Department of Plant Breeding and Seed Technology, University of Agriculture, Abeokuta Ogun State, Nigeria

Abstract A survey was conducted in 2000 to determine the incidence of Fusarium spp and
fumonisin B1 contamination in maize in farm stores in eight areas of Ogun State, Nigeria.
F. moniliforme was the predominant species, in 56% of the samples, with a mean kernel
infection of 68% in positive samples. The other species in decreasing order of abundance
were F. graminearum, F. pallidoroseum, F. solani and F. oxysporum. Analysis by high
performance liquid chromatography (HPLC) indicated that 51% of the samples were conta-
minated with fumonisin B1, at 65 to 1830 µg/kg.

Keywords: maize, Fusarium spp., fumonisin B1, storage.

Introduction
Fungi cause kernel rots of maize in the field and in storage, and Fusarium is the most
important genus of the field fungi. Strains of F. moniliforme and F. proliferatum produce
toxic metabolites known as fumonisins (Bacon and Nelson 1994). F. moniliforme is a
frequent contaminant of maize in Nigeria (Adesuyi and Shode 1977; Essien 2000), but infor-
mation on fumonisin in maize in West Africa is only available from Benin (Doko et al.
1995). This study was undertaken to identify Fusarium spp. and determine the levels of
fumonisin B1 in maize from farmers’ stores in Ogun State of Nigeria.

Materials and methods

Maize samples were collected from farm stores in eight areas (Table 1). The farmers stored
maize in outdoor bamboo cribs or in stacked polyethylene bags, or indoors in jute sacks or on
the floor of a hut. All the stored maize was meant for human consumption. For each
sampling, subsamples were collected from four points in each of three different layers of a
storage system giving a total of 12 sampling sites. Composite samples were obtained by
pooling and thoroughly mixing the subsamples. Sample sizes ranged from 500 g to 1 kg
depending on the capacity of the store. About half of each composite sample was milled and
a 10 g portion was used for analysis. The proportion of visibly mouldy, rotted or discoloured
kernels was determined on 200 representative kernels from each sample (Bankole et al.
1999).

Accepted 12 July 2002

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Table 1. Number of maize samples contaminated with Fusarium spp. in eight areas of Ogun State

Area Imeko Ipokia Odeda Owode Yewa – Abk – Ikenne Ijebu – Total
North North North samples

Total samples 21 16 10 8 18 8 12 15 108

F. oxysporum 0 2 0 1 0 1 2 2 08
F. moniliforme 13 7 6 5 8 4 7 10 60
F. solani 2 3 0 1 3 2 0 1 12
F. graminearum 8 1 3 2 4 3 2 5 28
F. pallidoroseum 5 4 1 2 4 0 3 2 11
Other Fusarium spp. 2 1 1 0 2 1 2 1 10
No Fusarium spp. 0 4 2 1 4 0 3 2 16

Fusarium spp. were isolated by directly plating kernels on peptone-pentachloroni-

trobenzene selective medium (Nash and Synder 1962). Ten kernels randomly picked from each
sample were surface-sterilized in 1% NaOCl for 1 min, and then rinsed in three changes of
sterile distilled water. The kernels were then plated on the agar plates (five grains/plate) and
incubated at 28 ± 2ºC with a 12 h photoperiod under Phillips fluorescent tubes for 5–10 days.
The colonies that grew from the kernels were transferred individually to potato dextrose agar,
incubated at 28 ± 2ºC for 10 days and identified by the method of Nelson et al. (1983).
Extraction, clean up and assay of fumonisin in maize samples were by a modification of the
method of Shephard et al. (1990). The extract obtained by stirring 10 g of ground maize in 20 ml
of methanol/water (3:1, v/v) for 30 min was filtered through Whatman 2V filter paper. The
filtrate (5 ml) was cleaned up by chromatography using a Bond-Elut SAX cartridge. The
cartridge was washed with 3 ml of methanol/water (4:1), and the toxin was eluted with 7:3
methanol:water. The eluate was evaporated to dryness under a gentle stream of nitrogen and kept
at 4ºC in a refrigerator until use. The final extracts were derivatized with o-phthaldialdehyde
solution, and the product of this reaction was analysed using a reverse-phase isocratic HPLC
system with an RF55 fluorescent detector (excitation wavelength 355 nm, emission wavelength
440 nm). The mobile phase was methanol/sodium acetate buffer (7:3 v/v, pH 3.6). The detection
limit for fumonisin B1 in this experiment was 50 µg/kg. Standard fumonisin B 1 was purchased
from the Council for Scientific and Industrial Research, South Africa.

Results and discussion

Fusarium contamination was found in 92 samples (85%; Table 1). F. moniliforme was the
most frequent species, in 56% of the samples. It predominated in all eight areas. The
percentage of kernels infected by F. moniliforme in positive samples ranged from 32 to 86%,
with a mean of 68%. F. graminearum was the second most prevalent, and F. pallidoroseum,
the third. F. solani and F. oxysporum were also isolated. The predominance of F. monili-
forme confirms the findings of Broadbent et al. (1969), Adesuyi and Shode (1977) and
Oyeniran (1977), that this was the most frequently occurring Fusarium on stored maize in
Trop Sci 43.2_crc 12/5/03 12:47 pm Page 78

78 SA Bankole et al.

Nigeria. Bankole (1994) reported that F. moniliforme constituted 57% of the fungi isolated
from freshly harvested maize in Nigeria.
Fumonisin B1 was detected in 55 of the 108 samples (Table 2). The proportion of positive
samples ranged from 40% in Ipokia and Odeda, to 67% in Ijebu North. The concentration of
fumonisin B1 varied from 65 to 1830 µg/kg with mean concentrations in positive samples of
390 µg/kg. Linear correlation analysis showed a significant positive correlation between the
number of samples positive to fumonisin B1 and those infected by F. moniliforme (P < 0.05;
r = 0.43). In South Africa, Rheeder et al. (1992) obtained fumonisin B1 concentrations of 1400
and 1600 µg/kg in high-quality and mouldy maize respectively from smallholder farms, while
Doko et al. (1995) reported 640 and 410 µg/kg in maize from Benin and Zambia respectively.
Kedara et al. (1999) found that maize from smallholder farms in Kenya had a mean level of
fumonisin B1 of 672 µg/kg. Thus the mean level of fumonisin B 1 in this study was lower than
the values in the other African countries except Zambia. Considering the high incidence of
F. moniliforme in maize, the levels of fumonisin B1 are lower than expected. A low correlation
between the incidence of F. moniliforme and fumonisin B1 was observed in maize in South
Africa (Munkvold and Desjardins 1997; Rheeder et al. 1995).

Table 2. Fumonisin B1 (µg/kg) in stored maize

Area % of samples No of samples Concentration of fumonisin B1

contaminated with positive for Mean* Range
F. moniliforme fumonisin (%)*

Imeko 62 13 (62) c 500 a 130–1830

Ipokia 44 08 (50) bc 350a 120–1130
Odeda 60 04 (40) b 440 b 130–1800
Owode 63 04(50) bc 390 ab 80–650
Yewa – North 44 10 (56) c 260 a 65–680
Abeokuta North 50 05 (63) c 580 bc 170–1600
Ikenne 58 03 (25)a 210 a 130–310
Ijebu – North 67 08 (53) bc 320 ab 90–1300
Total 56 55 (51) 390 65–1830

*Figures followed by the same letter(s) are not significantly different (P ≤ 0.05) by the Chi square test.

On the disease ratings of maize samples, 35 were in quality grade 1 (1–10% diseased
kernels), 41 were grade 2 (10–20%) and 17 were in grade 3 (21–30% diseased kernels) while
15 were in grade 4 (> 30%diseased kernels). Fourteen samples in grade 1 contained fumonisin
B1 as did 26 of grade 2, 11 of grade 3 samples and only four of grade 4 samples. Thus the extent
of visible mouldiness could not be used to assess fumonisin B1 contamination of stored maize

Acknowledgements
We thank the International Foundation for Science, Sweden, and the Organisation of Islamic
Conference Standing Committee on Scientific and Technological Cooperation, Pakistan, for
financial help.
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