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Research Article Open access CC BY 4.0

Prevalence of Aflatoxin Biosynthesis Genes According to Aflatoxin Levels in Maize of Different Varieties in Kenya

Samson Chebon, Wanjiru Wanyoike, Christine Bii, James Gathumbi, Dorington Ogoyi

Biotechnology Journal International · pp. 1–21 · Published 11 Aug 2017

10.9734/BJI/2017/28685

Abstract

Aims: To determine aflatoxin biosynthetic genes in fungal isolates in relation to aflatoxin levels in maize grain varieties from epidemiologically aflatoxicosis hot spots and non-aflatoxin hot spots agro-ecological zones in Kenya. Study Design: Purposeful sampling technique was applied targeting regions of variable aflatoxicoses susceptibilities. Place and Duration of Study: Samples were sourced from Kitui/Kibwezi counties, an aflatoxin hot spot, during 2008-2010 Maize growing seasons. Comparative samples were from Uasin-Gishu county and Perkerra irrigation scheme in Baringo County, both with no previous acute aflatoxicosis and practicing commercial cultivation under rain-fed and irrigation farming systems, respectively. Methodology: Maize samples (n=295) and fungal isolates (n=61) were analyzed for aflatoxin contamination and presence of aflatoxin biosynthetic genes, respectively. Total aflatoxin quantification was by a commercial Enzyme Linked Immunosorbent Assay (ELISA) kits, Boratest®, while molecular characterization of Aspergillus flavus (n=40) and A. parasiticus (n=21) isolates applied Quadruplex Multiplex PCR technique encompassing four aflatoxin biosynthetic genes: nor-1, ver-1, omt-A and aflR. Findings from the study variables were analyzed according to maize variety, agro-ecological origin of maize samples and fungal species besides type of farming system. Results: Uasin-Gishu maize samples (n=158) assayed for aflatoxins belonged to six maize commercial varieties; H614, H629, H6213, H6210, H613 and H628 alongside an indigenous variety, Kipkaa. All Perkerra samples (n=61) also belonged to a commercial variety, H513. Contrastingly, all Kitui/Kibwezi samples (n=76) belonged to an indigenous variety, Kikamba (Kinyanya). The varieties H613, H628 and Kipkaa all had the samples (100%) within the Kenyan statutory safe total aflatoxin limit (≤10.0 ppb) whereas Kikamba and H513 varieties had 82.9% and 83.6% samples within safety limits, respectively. Similarly, the mean aflatoxin content for all the seven Uasin-Gishu varieties was only 1.62 ppb while Kikamba and H513 had means of 14.6 ppb and 15.6 ppb, respectively (P=0.05). Positive PCR amplification results were obtained in 96.3%, 84.2% and 80% for Kitui/Kibwezi, Perkerra and Uasin-Gishu isolates, respectively whereas regional distribution of amplicon spectrum was 6, 3 and 2 out of 8, respectively. A similar regional pattern was established regarding prevalence of PCR positive isolates and whose maize samples of origin also tested ELISA-aflatoxin positive, having been 81.5%, 52.6% and 26.7% for Kitui/Kibwezi, Perkerra and Uasin-Gishu. Interestingly, the only isolate PCR positive for all the four genes under assay and whose maize sample of origin had aflatoxins was coincidentally from Kitui/Kibwezi, an epidemiologically aflatoxicosis  hot spot agro-ecological zone.

Multiplex PCR aflatoxin biosynthesis genes maize varieties Kenya

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