Food Control 22 (2011) 1760e1764

Contents lists available at ScienceDirect

Food Control
journal homepage: www.elsevier.com/locate/foodcont

Natural occurrence of Aflatoxin B1 in peanut collected from Kinshasa,

Democratic Republic of Congo
I. Kamika a, b, *, Losona L. Takoy a
a
Department of Biology, Faculty of Sciences, University of Kinshasa (UNIKIN), PO Box 190, Kinshasa XI, The Democratic Republic of the Congo
b
Department of Life Science, School of Agriculture and Environmental Sciences, University of South Africa (UNISA), PO Box 3037, Pretoria 0001, Gauteng, South Africa

a r t i c l e i n f o a b s t r a c t

Article history: Aflatoxin B1 is a potent carcinogen to both animal and human health. Since peanut is a suitable substrate
Received 24 July 2010 for aflatoxin production as well as an important oilseed and food in the Democratic Republic of Congo,
Received in revised form the risk of consuming aflatoxin-contaminated peanuts is very high. This paper assessed the natural
2 April 2011
occurrence of aflatoxin B1 in raw peanuts collected in rural areas of Kinshasa, Democratic Republic of
Accepted 9 April 2011
Congo. A total of 60 peanut samples were analyzed for aflatoxin B1, using thin layer chromatography. The
results show that aflatoxin B1 levels increased from the dry season to the rainy season with values
Keywords:
ranging from 1.5 to 390 and 12 to 937, respectively. 70% of the peanut samples from both seasons
Mycotoxin
Aflatoxin B1
exceeded the maximum limit of 5 mg/kg prescribed by the World Health Organization (WHO). As the
DRC Democratic Republic of Congo is amongst African countries listed with high prevalence of liver cancer,
Peanut continuous research on aflatoxin B1 is sought after.
Fungus Ó 2011 Elsevier Ltd. All rights reserved.
Aspergillus

1. Introduction Njapau, Muzungaile, & Changa, 1998). Stable once in foods,

Fungi are ubiquitous in the natural environment and can grow on
almost any organic matter (Marquardt, 1996). These fungi produce
diverse metabolites, mycotoxins that occur as both food and animal
feed contaminants. Among these fungi, Aspergillus, Fusarium and
Penicillium species are the most prevalent in food-crop as they can
contaminate in the field, during harvest, after harvest and during
storage (Bankole & Adebanjo, 2003). While Aspergillus and Penicil-
lium species are commonly found in stored grain or food, and
produce aflatoxins, ochratoxins, and citrinin, respectively (JECFA,
2001; Marquardt, 1996). The appearance of mycotoxins is as old as
agriculture itself and their effects were even represented in Egyp-
tian hieroglyphics (Dutton, 1990). However they received scientific
attention in 1960 when 100,000 turkey poults died after consuming
aflatoxin-contaminated peanut meal (Blount, 1961).
Because of their ability to contaminate human food and animal
feeds, aflatoxins have become the most prevalent mycotoxin due to
their economic and health significance (Bhat & Vashanti, 1999;
* Corresponding author. Department of Life and Consumer Sciences, School of
Agriculture and Life Sciences, College of Agriculture and Environmental Sciences,
University of South Africa (UNISA), PO Box 3037, Pretoria 0001, Gauteng, South
Africa. Tel.: þ27 72 7374336; fax: þ27 12 320 8168.
E-mail address: alainkamika2@yahoo.com (I. Kamika).
aflatoxins are naturally occurring mycotoxins found in maize,
peanut, cottonseed, spices and other feed grains (McDonald &
Harkness, 1964; Phillips, 1997); but mostly in peanut and maize
(Bankole & Adebanjo, 2003). Unfortunately peanut, as a suitable
substrate for fungal growth and aflatoxin productions, is an
important oilseed and food for many Congolese. Moreover, D’Mello
(2003) reported that aflatoxins are the most significant problem
regarding the quality of peanuts worldwide. It is estimated that 25%
of the world’s food commodities are contaminated by mycotoxins
each year (Magan, Sanchis, & Aldred, 2004).
Like many microbial secondary metabolites, aflatoxins are
a family of closely related compounds which include aflatoxin B1,
B2, G1 and G2, however aflatoxin B1 is usually in the highest
concentration and the most toxic (Henry, Bosch, Troxell, & Bolger,
1999; Lewis et al., 2005). According to several experimental data
and epidemiologic studies in the human population, aflatoxins,
particularly aflatoxin B1 (AFB1), is a potent carcinogen to humans
and classified as human carcinogen group 1 by the International
Agency for Research on Cancer (Hussein & Brasel, 2001; IARC, 1993;
Moss, 1996). Many authors confirmed that the liver is the primary
target organ of aflatoxins such as AFB1 (Hendrickse, 1983; Van
Rensburg et al., 1985). It also affects other organ systems leading
to diverse illness (Ciegler & Bennett, 1980; Coulombe, 1994;
Dvorackova, Stora, & Ayraud, 1981; Fung & Clark, 2004; Gong
et al., 2002; Qureshi, Brake, Hamilton, Hagler Jr., & Nesheim, 1998).

0956-7135/$ e see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodcont.2011.04.010
 I. Kamika, L.L. Takoy / Food Control 22 (2011) 1760e1764
AFB1 is known as a potent hepatocarcinogen and can play
a synergic action with hepatitis B or C viral infections leading to
hepatocellular carcinoma and also with other mycotoxins like
ochratoxins and fumonisin (Li, Yoshizawa, Kawamura, Luo, & Li,
2001; McKean et al., 2006; Parkin, Laara, & Muir, 1988; Turner
et al., 2000, Turner, Moore, Hall, Prentice, & Wild, 2003). It is
a serious health and economic problem worldwide, and pushed
many countries, and international agencies to establish the legis-
lation for maximum limits of mycotoxins, especially AFB1 in foods
for human and animal consumption (Creppy, 2002; Moss, 2002).
The United Nations-Food and Agriculture Organization (FAO) as
well as the World Health Organization (WHO) were faced with the
dilemma of setting the limits for aflatoxin in human foods against
high background levels of malnutrition; including the danger that
aflatoxin could produce liver cancer. They established the upper
aflatoxin limits of 30 mg/kg in foods for human consumption (Moss,
1996).
Recently, the European Union fixed the stringent maximum
residue levels for total aflatoxin and AFB1 levels in human
commodities to 4 and 2 mg/kg respectively (Moss, 2002), which had
an important impact on the economy of several African countries
(Dimanchie, 2001; Otzuki, Wilson, & Sewadeh, 2001). The Codex
Alimentarius commission (joint FAO/WHO) also adopted the limit
for total aflatoxin at 15 mg/kg in peanut (Codex, 2001). It should
nevertheless be mentioned that the WHO prescribed the maximum
limit for AFB1 in various foodstuffs at 5 mg/kg (Papp, H-Otta, Zaray,
& Mincsovics, 2002).
Exposure to aflatoxin is the most regulated and surveyed of all
the mycotoxins over the world, especially in more economically
developed parts of the world (Shephard, 2003; Van Egmond, 1989),
while, in developing countries, such protection is absent because of
high poverty (Ramos & Hernández, 1996). Unfortunately, factors
inducing fungal spoilage and aflatoxins productions are more
present in developing countries than elsewhere (Kamika, 2005).
Several authors have affirmed that aflatoxin problems are more
serious in developing countries such as the Democratic Republic of
Congo (DRC) where the climatic conditions (water activity or
moisture and temperature), transportation, marketing, and storage
practices are inadequate and considered conducive to fungal
growth and mycotoxin productions (Moss, 1996; Nakai et al., 2008;
Papp et al., 2002).
The DRC has an average temperature of 25.2  C with the highest
average monthly temperature (31  C) in February, March, April and
May. The average monthly relative humidity ranges from 74% in
August & September to 83% in January, November and December
(Brudzynski, 1985; Kazadi & Kaoru, 1996.) which are very close to
optimum conditions for Aspergillus growth as reported by Hell,
Cardwell, Setamou, and Poehling (2000).
The effects of aflatoxins, food safety and food security are
becoming a great threat to the health of both human and animal in
Africa (Shephard, 2003). Moreover, a high incidence of aflatoxicosis
was reported in Sub-Saharan countries (i.e. Kenya, Mozambique)
and India as well (Ciegler & Bennett, 1980; Lewis et al., 2005). In
DRC, the humanitarian crisis remains among the most complex,
deadly and prolonged ever documented since the World War 2
(Coghlan et al., 2006). This resulted in massive socio-economic
hazards, food crisis and malnutrition, and unfortunately there is
not enough data on food safety and food security. Indeed, Wagacha
and Muthomi (2008) estimated that there is a correlation between
the socio-economic status of the majority of sub-Saharan countries
and exposure to mycotoxins. The DRC can be classified among the
countries without permissive maximum levels of aflatoxins
(Shephard, 2003; Van Egmond, 1989). In 1977, Brudzynski, Van Pee,
and Kornazewski (1977) recorded the presence of aflatoxins up to
1000 mg/kg in many foods such as maize and peanuts from the DRC
1761
(former Zaire) and emphasized that the DRC has favorable climatic
conditions for fungal growth and the rural population may be the
most exposed to aflatoxins.
Additionally, the presence of aflatoxins, especially AFB1 in
peanuts and peanut products, were also found elsewhere due to the
peanut’s suitability for fungal growth and aflatoxins production
(Barro, Quattara, Nikiema, Quattara, & Traore, 2002; Brigido, 1998;
Garcia, 1989; McDonald, 1964; Mutegi, Ngugi, Hendriks, & Jones,
2009; Ominski, Marquardt, Sinah, & Abramson, 1994; Siame,
Mpuchane, Gashe, Allotey, & Teffera, 1998).
As the DRC is known to have favorable climatic conditions for
fungal growth and classified on the list of African countries with
a high risk of primary liver cancer (Stora, 1990), the purpose of this
study was to assess AFB1 level in raw peanuts collected during both
rainy and dry seasons in the DRC.
2. Materials and methods
2.1. Chemicals and reagents
All chemicals used were of analytical-grade and were purchased
from Sigma and Merck (Pretoria, PTA, RSA). Aflatoxin B1 standard
was purchased from Supelco (Bellefone, PA, USA).
2.2. Samples
Sixty peanut samples were purchased from street hawkers, local
markets and retail shops in rural areas of Kinshasa (Matadimayo,
Kinseso, Kimwenza, Djili Secomaf, Djili Brasserie), DRC. The
samples were collected randomly in the year 2009 during the dry
season (May and June) and rainy season (October and November).
They were dispensed into plastic bags and stored at 4  C until
analysis.
2.3. Aflatoxin B1 analysis
This analysis was conducted at the Quality Control Laboratory of
the “Office Congolais de Control” (OCC) and at the University of
Kinshasa, DRC.
2.3.1. Standard preparation
The standard solution was prepared according to AOAC-971.22
method (AOAC, 1990). Briefly, the solid standard (Sigma, Sigma
Chemical Co - St. Louis, MO) was dissolved in benzene/acetonitrile
(98/2) and vigorously agitated for 1 min on shaker (Flask Shaker SF
1, Stuart scientific, Poole, UK). The purity chromatographic and the
exact concentration were determined by spectrophotometer
(PerkineElmer, USA). A working solution of 0.55 mg/mL was
prepared by diluting appropriate aliquots of stock solution in
benzene/acetonitrile (98/2) and stored at 20  C.
2.3.2. Extraction of aflatoxin B1
The extraction of AFB1 was carried out according to the
AOAC-970.45D method (AOAC, 1990) slightly modified. Briefly,
100 g of prepared samples of raw peanuts were mixed with 350 ml
methanol/water (85/15), 200 ml of hexane and 4g of sodium
chloride were added and blended for 3 min at high speed by High
speed blender (Waring Products DIV., Torrington, CT). After
blending, samples were immediately filtered through filter paper
(Whatman No.1) and 250 ml of the filtrate was centrifuged for
5 min at 2000 rpm.
25 mL of aqueous methanol phase was pipetted into a 125 mL
separating funnel, mixed with 25 mL chloroform (CHCl3) and
agitated for 30e60 s. The layers separation was let and drained the
bottom layer of the CHCl3 through anhydrous sodium sulfate into
1762 I. Kamika, L.L. Takoy / Food Control 22 (2011) 1760e1764

stainless steel beaker of 600 mL and repeated twice with 25 mL Table 1

portions of chloroform. The beaker was placed on the steam plate Recovery test of AFB1 in raw peanut samples.

under stream of nitrogen (N) and evaporated to dryness and dis- Sample Number AFB1 level AFB1 recovered %Recovery
solved in 5 mL methanol/water (85/15). The extract was diluted of Analyses added (mg/kg) (mg/kg) (mean  SD) (mean  SD)
with 20 ml of phosphate-buffered saline (PBS) solution and then Peanut 3 2 1.63  0.31 75.14  12.53
filtered using glass microfiber filter. Peanut 3 4 3.70  0.28 82.23  13.18
Peanut 3 6 5.49  0.16 87.63  9.04
Filtrate (20 mL) was passed through an immuno-affinity column
Peanut 3 10 8.94  0.09 71.50  5.04
(IAC, Rhone-Poulenc Diagnostics, UK) previously conditioned in
PBS at a flow-rate of 1e2 drops/seconds. The IAC was washed with
15 mL of 0.1% Tween-20 followed with 10 mL of distilled water until
air came through the column. AFB1 was eluted with 2 mL of HPLC 2003; Barro et al., 2002; Brudzynski et al., 1977; Garcia, 1989;
grade methanol at a rate of 1e2 drops/seconds and eluate collected McDonald, 1964; Mutegi et al., 2009).
in screw capped borosilicate vial. The eluate was well mixed with In this study, thin layer chromatography was used to determine
50 mL of methanol-formic acid solution, taken to dryness at 40  C AFB1 level in peanut samples collected during three months of
under gentle stream of N and re-dissolved in 200 mL of bezene/ rainy season and three months of dry season from different loca-
acetonitrile (98/2) for spotting on thin layer chromatography (TLC) tions. The limit of the detection of AFB1 was 1 mg/kg and the
plate (Stroka, van Otterdijk, & Anklam, 2000). recovery values ranged from 71% to 87% (see Table 1). These
recovery values were higher than the values obtained from raw and
2.3.3. Aflatoxin B1 determination on TLC roasted peanuts with TLC detection method conducted by Kamika
The identification and quantification of AFB1 was performed by (2005) which ranged from 56% to 72%.
thin layer chromatography (AOAC 968.22F(c)-(d)) on silica gel 60 Table 2 summarizes the results of peanut samples collected
plates of dimension 20  20 cm (Merck, Art.5553) according to during rainy and dry seasons. 27 peanut samples, corresponding to
AOAC (1990). After reconstitution of sample to 200 mL benzene/ 90% of peanut samples collected during the rainy season were
acetonitrile (98/2) and homogenization, 5 mL of sample was spotted contaminated with AFB1 at a concentration ranging from 12 to
on a plate along with a working standard solution. The plate was 937 mg/kg with a mean of 205.7 mg/kg. In November all the samples
first eluted with anhydrous ethyl ether, dried up in a fume hood for were positive and contained the highest concentration of AFB1 937
5 min, and developed with chloroform/acetone (90/10) in the same mg/kg. While in the dry season peanut samples were contaminated
direction. The TLC plate was visually examined under ultraviolet with AFB1 in only 16 peanut samples, corresponding to 53% of the
(UV) light at 366 nm. The AFB1 level in the samples was calculated peanut samples collected and analyzed during the dry season.
by comparing the area of chromatographic peak of the samples The samples contained AFB1 concentrations ranging from 1.5 to
with the AFB1 standard solution by densitometric analysis 390 mg/kg with the highest AFB1 level at 390 mg/kg in June.
(Shimadzu Densitometer, CS9301PC, Shimadzu Scientific Instru- The highest level of AFB1 for both seasons was detected in
ments, Japan) 200e300 nm, mercury lamp l ¼ 366 nm, photo samples collected during the rainy season (November: 937 m/kg).
mode: fluorescence, beam size: 0.4  5.0 mm, reference wave: On the contrary, the lowest level of AFB1 was detected in May
360 nm, scan mode: linear, fluorescence sensitivity: high. (1.5 m/kg) during the dry season.
Several studies investigated the presence of aflatoxins, partic-
3. Recovery detection ularly AFB1, in human foods such as peanuts. Brudzynski et al.
(1977) found AFB1 in many Congolese foods including peanuts,
To test the sensitivity of the method, the AFB1 standard solution maize and cassava ranging from 12 to up to 1000 mg/kg. Another
at different concentrations (2, 4, 6, and 10 mg/kg) were added to the investigation reported the presence of aflatoxin at a concentration
peanut samples suspected to contain less than 1 mg/kg of aflatoxin. up to 2000 mg/kg in Nigerian peanut (McDonald, 1964) and the
The extraction of the spiked peanuts was done as described above, presence of aflatoxin ranging from 12 to 329 mg/kg in peanut
in triplicate. samples collected from Botswana (Barro et al., 2002).
The recovery of AFB1 from spiked peanut samples was deter- Recently, Mutegi et al. (2009) also analyzed the prevalence and
mined as previously described. factors associated with aflatoxin contamination of peanuts from

4. Data analysis

Data were analyzed using the SPSS program. Tests of signifi- Table 2
cance and relationships were carried out using Analysis of variance Presence of AFB1 in peanut samples collected during Dry and Rainy seasons.

(ANOVA) and linear regression models, respectively. Unless indi- Months Samples Positive Range Mean Frequency
cated otherwise, tests for significance were performed at the 5% analyzed samples No (m/kg) (m/kg) distribution of
level. No (%) AFB1 level (%)

5 m/kg >5 m/kg

5. Result and discussion Rainy season
October 10 9 (90%) 12e370 94.7 10% 90%
In the DRC, peanuts are consumed raw, cooked (roasted or November 10 10 (100%) 67e937 310.4 0% 100%
December 10 8 (80%) 15e910 212.1 20% 80%
boiled) or mixed with other food substances (vegetables, meats, Sub-total 30 27 (90%) 12e937 205.7 10% 90%
fishes, etc.) and considered as an everyday meal by many rich and Dry season
poor Congolese families. The aflatoxin contamination problems May 10 4 (40%) 1.5e50 9.45 80% 20%
have also been encountered in different foods consumed by the June 10 7 (70%) 3.5e390 60.85 40% 60%
July 10 50 (50%) 2e80 14.8 60% 40%
population (Brudzynski et al., 1977). Since peanut is considered to
Sub-total 30 16 (53%) 15e390 23.37 50% 50%
be one of the most susceptible food materials for fungal growth and
aflatoxin productions, several researchers have investigated it for Total 60 43 (72%) 1.5e937 229.07 30% 70%

the presence of aflatoxins, particularly AFB1 (Bankole & Adebanjo, No: Number.
 I. Kamika, L.L. Takoy / Food Control 22 (2011) 1760e1764
western Kenya and found that peanut samples from both regions
were contaminated and aflatoxin ranged from ND to 7525 mg/kg.
The present study revealed that the occurrence of AFB1 in both
seasons increased from the dry season to the rainy season and this
could be explained in terms of climatic conditions as reported by
Hell et al. (2000).
In addition, this study is in agreement with other previous
studies on the natural occurrence of AFB1 in peanut samples from
other countries, due to the susceptibility of peanuts to be
contaminated by aflatoxins in favorable climatic conditions
(Nakai et al., 2008).
Our result also shows that AFB1 detected in peanut samples
collected in both rainy and dry seasons had mean values of
205.70 mg/kg and 23.37 mg/kg, respectively. Approximately 72% of
the peanut samples were positive (see Table 2) and 70% higher than
the maximum limit prescribed by WHO at 5ug/kg for AFB1 (Papp
et al., 2002). 95% of the positive samples from both seasons
appeared to contain AFB1 above the European standard (2 mg/kg) in
peanuts intended for direct human consumption, whereas all the
positive samples collected during the rainy season were found with
an unacceptable concentration AFB1 for human consumption.
6. Conclusion
This study revealed the occurrence of aflatoxin B1 in raw
peanuts collected from the Democratic Republic of Congo. The
occurrence of this AFB1 increased from the dry season to the rainy
season while the highest level of AFB1 was observed in the samples
collected during November.
Approximately 72% of the peanut samples analyzed were posi-
tive and 70% exceeded the maximum limit of 5 mg/kg prescribed by
WHO (Papp et al., 2002). This is an alarming indication especially
since the DRC does not have an existing regulation on mycotoxins
in food.
However, it should be emphasized that continuous study, with
a greater number and variety of food commodities from different
locations, should be performed to evaluate the natural occurrence
of aflatoxins countrywide. This was a preliminary study on
aflatoxins implemented in Kinshasa, DRC since 1977.
Acknowledgment
The authors are grateful to the Office Congolais de Control (OCC),
Kinshasa, Democratic Republic of Congo for allowing unrestricted
use of their laboratory facilities. We also thank Dr. Muller Kabamba
for his technical assistance.
References
Association of Official Analytical Chemists (AOAC). (1990). Official methods of
analysis (15th ed). Virginia, USA.
Brigido, B. M. (1998). Occurrence of aflatoxins B1, B2, G1, and G2 in peanuts and their
products marketed in the region of Campinas, Brazil in 1995 and 1996. Food
Additives & Contaminants, 15(7), 807e811.
Bankole, S. A., & Adebanjo, A. (2003). Mycotoxins in food in West Africa: current
situation and possibilities of controlling it. African Journal of Biotechnology, 2(9),
254e263.
Barro, N., Quattara, C. A., Nikiema, P. A., Quattara, A. S., & Traore, A. S. (2002).
Microbial quality assessment of some street food Widely consumed in Ouaga-
dougou, Burkina Faso. Sante, 12, 369e374.
Bhat, R. V., & Vashanti, S. (1999). Occurrence of aflatoxins and its economic impact
on human nutrition and animal feed. The New Regulation. Agricultural Devel-
opment, 23, 50e56.
Blount, W. P. (1961). Turkey ‘X’ disease. Journal of British Turkey Federation, 9, 52e61.
Brudzynski, A., Van Pee, W., & Kornazewski, W. (1977). The occurrence of aflatoxin
B1 in peanuts, corn and dried cassava sold at the local market in Kinshasa,
Zaire: its coincidence with high hepatoma morbidity among the population.
Zeszyty Problemowe Postepow Nauk Rolniczych, 189, 113e115.
Brudzynski, A. (1985). L’incidence d’Aspergillus flavus et d’autres espèces de moi-
sissures dangereuses dans la mycoflore du mais, d’arachides et de cossettes du
1763
manioc vendues sur le marche de Kisangani. Industries Alimentaires et Agricoles,
102(4), 307e309.
Ciegler, A., & Bennett, W. (1980). Mycotoxins and Mycotoxicoses. BioScience, 30(8),
512e515.
Codex Alimentarius Commission. (2001). Joint FAO/WHO food standards programme,
codex committee on food additives and contaminants. Thirty-third session. Hague,
Netherlands: CODEX.
Coghlan, B., Brennan, R., Ngoy, P., Dofara, D., Otto, B., Clements, M., et al. (2006).
Mortality in Democratic Republic of Congo: a nationwide survey. Lancet, 367,
44e51.
Coulombe, R. (1994). Nonhepatic disposition and effects of aflatoxin B1. In D. Eaton,
& J. Groopman (Eds.), The Toxicology of aflatoxins: Human health, veterinary, and
agricultural significance (pp. 89e101). San Diego: Academic Press.
Creppy, E. E. (2002). Update of survey, regulation and toxic effects of mycotoxins in
Europe. Toxicology Letters, 127, 19e28.
Dimanchie, P. (2001). Groundnut exporters in Southern countries penalized by new
standards on aflatoxins imposed by the European Union. OCL - Oleagineux,
Corps Gras, Lipides, 8, 237e238.
D’Mello, J. P. F. (2003). Mycotoxins in cereal grains, nuts and other plant products. In
J. P. F. D’Mello (Ed.), Food safety: Contaminants and toxins (pp. 65e90). Wall-
ingford: CAB International.
Dutton, M. F. (1990). Mycotoxins and other natural causes of deaths. Pieterrmaritz-
burg: University of Natal Press, SA.
Dvorackova, I., Stora, C., & Ayraud, N. (1981). Evidence for aflatoxin BI in two cases
of lung cancer in man. Cancer Research and Clinical Oncology, 100, 221e224.
Van Egmond, H. P. (1989). Current situation on regulations for mycotoxins. Over-
view of tolerances and status of standard methods of sampling and analysis.
Food Additives & Contaminants, 6(2), 139e188.
Fung, F., & Clark, R. F. (2004). Health effects of mycotoxins: a toxicological overview.
Clinical Toxicology, 42(2), 217e234.
Garcia, V. V. (1989). An overview of peanut utilization in the Philippines. In
T. C. Yam, & C. Tan (Eds.), Trends in food product development (pp. 21e26).
Singapore: Singapore Institute of Food Science and Technology.
Gong, Y. Y., Cardwell, K. K., Hounsa, A., Eggal, S., Turner, P. C., Hall, A. J., et al. (2002).
Dietary aflatoxin exposure and impaired growth in young children from Benin
and Togo: a cross-sectional study. British Medical Journal, 325, 20e21.
Hell, K., Cardwell, K. F., Setamou, M., & Poehling, H. M. (2000). The influence of
storage practices on aflatoxin contamination in maize in four agroecological
zones of Benin, West Africa. Journal of Stored Products Research, 36, 365e382.
Henry, S. H., Bosch, F. X., Troxell, T. C., & Bolger, P. M. (1999). Reducing liver cancer-
global control of aflatoxin. Science, 286(5449), 2453e2454.
Hendrickse, R. G. (1983). Aflatoxin and kwashiorkor: epidemiology and clinical
studies in Sudanese children and findings in autopsy liver sample from Nigeria
and South Africa. Bulletin de la Société de Pathologie Exotique et de ses Filiales, 76,
559e566.
Hussein, H. S., & Brasel, J. M. (2001). Toxicity, metabolism, and impact of mycotoxins
on humans and animals. Toxicology, 167, 101e134.
IARC. (1993). Some naturally occurring substances: Food items and constituents,
heterocyclic amines and mycotoxins. IARC Monographs on the Evaluation of
Carcinogenic Risks to Humans, 56. Lyon, France, International Agency for
Research on Cancer.
Kamika, I. (2005). L’impact de la température et l’humidité sur la production
d’aflatoxins. Annal Universitaire (Universite de Kinshasa), 102e115.
JECFA (Joint FAO/WHO Expert Committee on Food Additives). (2001). Fifty-sixth
meeting, 6e15 February, Geneva.
Kazadi, S. N., & Kaoru, F. (1996). Interannual and long-term climate variability over
the Zaire River Basin during the last 30 years. Journal of Geophysical Research,
101(D16), 21,351e21,360.
Lewis, L., Onsongo, M., Njapau, H., Schuzz-Rogers, H., Luber, G., Kieskak, S., et al.,
the Kenya aflatoxicosis Investigation Group. (2005). Aflatoxin contamination of
commercial maize products during an outbreak of acute aflatoxicosis in Eastern
and central Kenya. Environmental Health Perspectives, 113, 1763e1767.
Li, F., Yoshizawa, T., Kawamura, O., Luo, X., & Li, Y. (2001). Aflatoxins and fumonisins
in corn from the high incidence area for human hepatocellular carcinoma in
Guangxi, China. Journal of Agricultural and Food Chemistry, 49, 4122e4126.
Magan, N., Sanchis, V., & Aldred, D. (2004). Role of spoilage fungi in seed deterio-
ration. In D. K. Aurora (Ed.), Fungal biotechnology in agricultural, food and
environmental applications, Chapter 28 (pp. 311e323). Marcell Dekker.
Marquardt, R. R. (1996). Effects of moulds and their toxins on livestock perfor-
mance: a western Canadian perspective. Animal Feed Science and Technology,
58(1e2), 77e89.
McKean, C., Tang, L., Tang, M., Billam, M., Wang, Z., Theodorakis, C. W., et al. (2006).
Comparative acute and combinative toxicity of aflatoxin B1 and fumonisin B1 in
animals and human cells. Food and Chemical Toxicology, 44(6), 868e876.
McDonald, D., & Harkness, C. (1964). Growth of Aspergillus flavus and production of
aflatoxin in groundnuts. Part IV. Tropical Science, 4, 12e27.
McDonald, D. (1964). Progress reports on research into aflatoxin production in
groundnuts in Northern Nigeria. Samaru Miscellaneous Papers, 2, 13.
Moss, M. O. (1996). Mycotoxic fungi. In A. E. Eley (Ed.), microbial food poisoning (2nd
ed) (pp. 75e93). London: Chapman & Hall.
Moss, M. O. (2002). Risk assessment for aflatoxins in foodstuffs. International
Biodeterioration & Biodegradation, 50, 137e142.
Mutegi, C. K., Ngugi, H. K., Hendriks, S. L., & Jones, R. B. (2009). Prevalence and
factors associated with aflatoxin contamination of peanuts from Western
Kenya. International Journal of Food Microbiology, 130(1), 27e34.
1764 I. Kamika, L.L. Takoy / Food Control 22 (2011) 1760e1764
Nakai, V. K., Rocha, L. O., Goncalez, E., Fonseca, H., Ortega, E. M. M., & Correa, B.
(2008). Distribution of fungi and aflatoxins in a stored peanut variety. Food
Chemistry, 106, 285e290.
Njapau, H., Muzungaile, E. M., & Changa, R. C. (1998). The effect of village processing
techniques on the content of aflatoxins in corn and peanuts in Zambia. Journal
of the Science of Food and Agriculture, 76, 450e456.
Ominski, K. H., Marquardt, R. R., Sinah, R. N., & Abramson, D. (1994). Ecological
aspects of growth and mycotoxin production by storage fungi. In J. D. Miller, &
H. L. Trenholm (Eds.), Mycotoxins in Grain-compounds other than Aflatoxin (pp.
287e312). St. Paul: Eagan Press.
Otzuki, T., Wilson, J. S., & Sewadeh, M. (2001). What price precaution? European
harmonisation of aflatoxin regulations and African groundnut exports. Euro-
pean Review of Agricultural Economics, 28, 263e283.
Papp, E., H-Otta, K., Zaray, G., & Mincsovics, E. (2002). Liquid chromatographic
determination of aflatoxins. Microchemical Journal, 73, 39e46.
Parkin, D. M., Laara, E., & Muir, C. S. (1988). Estimation of the worldwide
frequency of sixteen major cancers in 1980. International Journal of Cancer, 41,
184e197.
Phillips, T. D. (1997). Detection and Decontamination of aflatoxin-contaminated
food products. In J. H. Williams, D. G. Cummins, G. Hutto, & A. King (Eds.),
Impacts and scientific advances through collaborative research on peanut CRSP
(pp. 117e130). Geogia, GA: Griffin.
Qureshi, M. A., Brake, J., Hamilton, P. B., Hagler, W. M., Jr., & Nesheim, S. (1998).
Dietary exposure of broiler breeders to aflatoxin results in immune dysfunction
in progeny chicks. Poultry Science, 77(6), 812e819.
Ramos, A. J., & Hernández, E. (1996). In vitro aflatoxin adsorption by means of
a montmorillonite silicate. A study of adsorption isotherms. Animal Feed Science
and Technology, 62(2e4), 263e269.
Shephard, G. S. (2003). Aflatoxin and food safety: recent African perspectives. Toxin
Reviews, 22(2e3), 267e286.
Siame, B. A., Mpuchane, S. F., Gashe, B. A., Allotey, J., & Teffera, G. (1998). Occurrence
of aflatoxins, fumonisin B1, and Zearalenone in foods and feeds in Botswana.
Journal of Food Protection, 61(12), 1670e1673, (4).
Stora, C. (1990). Intracellular localization of aflatoxin B sub(1) in rat and human livers.
Journal of Environmental Pathology, Toxicology and Oncology, 10(3), 129e131.
Stroka, J., van Otterdijk, R., & Anklam, E. (2000). Immunoaffinity column clean-up
prior to thin-layer chromatography for the determination of aflatoxins in
various food matrices. Journal of Chromatography A, 904, 251e256.
Turner, P. C., Mendy, M., White, H., Fortuin, M., Hall, A. J., & Wild, C. P. (2000).
Hepatitis B infection and aflatoxin biomarker levels in Gambian children.
Tropical Medicine & International Health, 5, 837e841.
Turner, P. C., Moore, S. E., Hall, A. J., Prentice, A. M., & Wild, C. P. (2003). Modification
of immune function through exposure to dietary aflatoxin in Gambian children.
Environmental Health Perspectives, 111, 217e220.
Van Rensburg, S. J., Cook-Mozaffari, P., Van Schalkwyk, D. J., Van der Watt, J. J., et al.
(1985). Hepatocellular carcinoma and dietary aflatoxin in Mozambique and
Transkei. British Journal of Cancer, 51(5), 712e726.
Wagacha, J. M., & Muthomi, J. W. (2008). Mycotoxin problem in Africa: current
status, implications to food safety and health and possible management strat-
egies. International Journal of Food Microbiology, 124, 1e12.