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Aflatoxins are mycotoxins that can contaminate grains, legumes, and oil seeds. These toxic compounds are an especially serious problem in tropical and sub-tropical climates. The objective of this study was to raise awareness of aflatoxin exposure among primary school children in Shebedino woreda, southern Ethiopia, by measuring urinary aflatoxin M1 (AFM1). The study employed a cross-sectional design and systematic random sampling of children from eight schools in the district. The mean ± SD age of the children was 9.0 ± 1.8 years. Most (84.6%) households were food insecure with 17.9% severely food insecure. Urinary AFM1 was detected in more than 93% of the children. The median [IQR] concentration of AFM1/Creat was 480 [203, 1085] pg/mg. Based on a multiple regression analysis: DDS, consumption of haricot bean or milk, source of drinking water, maternal education, and household food insecurity access scale scores were significantly associated with urinary AFM1/Creat. In conclusion, a high prevalence of urinary AFM1 was observed in this study. However, the relation between AFM1 and dietary intake was analyzed based on self-reported dietary data; hence, all of the staple foods as well as animal feeds in the study area should be assessed for aflatoxin contamination.
The study was conducted in Shebedino woreda, Sidama Zone, SNNPR. Shebedino woreda, located in the Great Rift Valley, lies approximately 1710 m above sea level and is a tropical climate. The average temperature is between 18–25 °C. The area is characterized by seasonal and variable rainfall with average annual rainfall between 900–1100 mm [19]. The study population depends on staple foods such as maize, haricot beans, and enset (false banana). Most of the population depend on mixed farming where farmers support their livelihoods from crop production and animal husbandry [19]. This cross-sectional school-based study was conducted from May to June 2017 as part of a study examining prevalence of iodine deficiency. Study subjects were randomly selected using a single proportion sample size calculation formula. A sample size of 408 school age children was computed and allocated to eight randomly selected schools by population proportional to size. Students were randomly selected from all children 6–12 years of age in those schools as explained in further detail in our previously published manuscript [20]. Demographic and socio-economic characteristics of study participants were assessed using questions adapted from the Ethiopian Demographic and Health Survey 2011 report [21]. Food consumption patterns of households and children were estimated using a standardized food frequency questionnaire [22]. Household food insecurity was assessed using the Household Food Security Access Scale (HFIAS) developed by the Food and Nutrition Technical Assistance (FANTA) project of the United States Agency for International Development (USAID) [23]. Children’s dietary diversity score was assessed based on the FAO recommendation [24]. Urine samples were collected from each participant in a wet season (May to June). Short term AF exposure was assessed by analyzing urine samples for AFM1 using a commercially available enzyme-linked immunosorbent assay (ELISA) kit for quantitative determination of aflatoxin in urine (Helica Biosystems Inc., Santa Ana, CA, USA). Briefly, the kit contained a microwell plate in which all wells had been coated with an antibody with high affinity for AFM1. When added to a microwell, the AFM1 in the urine sample bound to the coated antibody. Next, a reagent containing aflatoxin bound to horse-radish peroxidase (HRP) was added to each well. After the incubation period, wells were decanted and washed with a PBS buffer with 0.05% Tween20. An HRP substrate was added to each well followed by a stop solution. The microwell plate was then read at 450 nm on a plate reader. Six standards provided with the kit established a standard curve from 0 to 200 pg/mL of aflatoxin. All standards and reagents were provided with the kit. In our hands, the limit of detection was 1.25 pg/mL of aflatoxin and the limit of quantification was 2.5 pg/mL. Urinary creatinine was analyzed using a BioLis 24i clinical chemistry analyzer with standard reagents (Carolina Liquid Chemistries Corp., Brea, CA, USA).
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