RESUMEN
Aflatoxins pose significant food security and public health risks, decrease productivity and profitability of animal industries, and hamper trade. To minimize aflatoxin contamination in several crops, a biocontrol technology based on atoxigenic strains of Aspergillus flavus is commercially used in the United States and some African countries. Significant efforts are underway to popularize the use of biocontrol in Africa by various means including incentives. The purpose of this study was to develop quantitative pyrosequencing assays for rapid, simultaneous quantification of proportions of four A. flavus biocontrol genotypes within complex populations of A. flavus associated with maize crops in Nigeria to facilitate payment of farmer incentives for Aflasafe (a biocontrol product) use. Protocols were developed to confirm use of Aflasafe by small scale farmers in Nigeria. Nested PCR amplifications followed by sequence by synthesis pyrosequencing assays were required to quantify frequencies of the active ingredients and, in so doing, confirm successful use of biocontrol by participating farmers. The entire verification process could be completed in 3-4 days proving a savings over other monitoring methods in both time and costs and providing data in a time frame that could work with the commercial agriculture scheme. Quantitative pyrosequencing assays represent a reliable tool for rapid detection, quantification, and monitoring of multiple A. flavus genotypes within complex fungal communities, satisfying the requirements of the regulatory community and crop end-users that wish to determine which purchased crops were treated with the biocontrol product. Techniques developed in the current study can be modified for monitoring other crop-associated fungi.
RESUMEN
Although recent reports indicated that produce contamination with foodborne pathogens is widespread in Nigeria, the sources and magnitude of microbial contamination of fruits and vegetables on farms and in markets have not been thoroughly identified. To ascertain possible pathways of contamination, the frequency and magnitude of coliform and Escherichia coli contamination of tomatoes produced in northwest Nigeria was assessed on farms and in markets. Eight hundred twenty-six tomato fruit samples and 36 irrigation water samples were collected and assessed for fecal indicator organisms. In addition, the awareness and use of food safety practices by tomato farmers and marketers were determined. Median concentration of coliforms on all field- and market-sourced tomato fruit samples, as well as in irrigation water sources, in Kaduna, Kano, and Katsina states exceeded 1,000 most probable number (MPN) per g. Median E. coli counts from 73 (17%) of 420 field samples and 231 (57%) of 406 market tomato fruit samples exceeded 100 MPN/g. Median E. coli concentrations on tomato fruits were higher (P < 0.01) in the rainy season (2.45 Log MPN/g), when irrigation was not practiced than in the dry (1.10 Log MPN/g) and early dry (0.92 Log MPN/g) seasons. Eighteen (50%) of 36 irrigation water samples had E. coli counts higher than 126 MPN/100 ml. Median E. coli contamination on market tomato fruit samples (2.66 Log MPN/g) were higher (P < 0.001) than those from tomatoes collected on farms (0.92 Log MPN/g). Farmers and marketers were generally unaware of the relationship between food safety practices and microbial contamination on fresh produce. Good agricultural practices pertaining to food safety on farms and in local markets were seldom used. Adoption of food safety practices on-farm, during transport, and during marketing could improve the microbial quality of tomatoes available to the public in this region of the world.