Assessing the mycotoxicological risk from consumption of complementary foods by infants and young children in Nigeria.

This study assessed, for the first time, the mycotoxicological risks from consumption of complementary foods by infants and young children in Nigeria. Molds belonging to Aspergillus aculeatinus, A. flavus, A. luchuensis, A. tubingensis, A. welwitschiae and Geotrichum candidum were recovered from the complementary foods. Twenty-eight major mycotoxins and derivatives, and another 109 microbial metabolites including chloramphenicol (a bacterial metabolite), were quantified in 137 food samples by LC-MS/MS. Aflatoxins and fumonisins co-contaminated 42% of the cereal- and nut-based food samples, at mean concentrations exceeding the EU limits of 0.1 and 200 µg/kg set for processed baby foods by 300 and six times, respectively. Milk contained mainly beauvericin, chloramphenicol and zearalenone. The trichothecenes, T-2 and HT-2 toxins, were quantified only in infant formula and at levels three times above the EU indicative level of 15 µg/kg for baby food. Chronic exposure estimate to carcinogenic aflatoxin was high causing low margin of exposure (MOE). Exposures to other mycotoxins either exceeded the established reference values by several fold or revealed low MOEs, pointing to important health risks in this highly vulnerable population. The observed mycotoxin mixtures may further increase risks of adverse health outcomes of exposure; this warrants urgent advocacy and regulatory interventions.

High-Throughput Sequence Analyses of Bacterial Communities and Multi-Mycotoxin Profiling During Processing of Different Formulations of Kunu, a Traditional Fermented Beverage.

Kunu is a traditional fermented single or mixed cereals-based beverage popularly consumed in many parts of West Africa. Presently, the bacterial community and mycotoxin contamination profiles during processing of various kunu formulations have never been comprehensively studied. This study, therefore, investigated the bacterial community and multi-mycotoxin dynamics during the processing of three kunu formulations using high-throughput sequence analysis of partial 16S rRNA gene (hypervariable V3-V4 region) and liquid chromatography tandem mass spectrometry (LC-MS/MS), respectively. A total of 2,303 operational taxonomic units (OTUs) were obtained across six processing stages in all three kunu formulations. Principal coordinate analysis biplots of the Bray-Curtis dissimilarity between bacterial communities revealed the combined influences of formulations and processing steps. Taxonomically, OTUs spanned 13 phyla and 486 genera. Firmicutes (phylum) dominated (relative abundance) most of the processing stages, while Proteobacteria dominated the rest of the stages. Lactobacillus (genus taxa level) dominated most processing stages and the final product (kunu) of two formulations, whereas Clostridium sensu stricto (cluster 1) dominated kunu of one formulation, constituting a novel observation. We further identified Acetobacter, Propionibacterium, Gluconacetobacter, and Gluconobacter previously not associated with kunu processing. Shared phylotypes between all communities were dominated by lactic acid bacteria including species of Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, and Weissella. Other shared phylotypes included notable acetic acid bacteria and potential human enteric pathogens. Ten mycotoxins [3-Nitropropionic acid, aflatoxicol, aflatoxin B1 (AFB1), AFB2, AFM1, alternariol (AOH), alternariolmethylether (AME), beauvericin (BEAU), citrinin, and moniliformin] were quantified at varying concentrations in ingredients for kunu processing. Except for AOH, AME, and BEAU that were retained at minimal levels of < 2 µg/kg in the final product, most mycotoxins in the ingredients were not detectable after processing. In particular, mycotoxin levels were substantially reduced by fermentation, although simple dilution and sieving also contributed to mycotoxin reduction. This study reinforces the perception of kunu as a rich source of bacteria with beneficial attributes to consumer health, and provides in-depth understanding of the microbiology of kunu processing, as well as information on mycotoxin contamination and reduction during this process. These findings may aid the development of starter culture technology for safe and quality kunu production.

Bacterial species and mycotoxin contamination associated with locust bean, melon and their fermented products in south-western Nigeria.

The microbiological safety of spontaneously fermented foods is not always guaranteed due to the undefined fermenting microbial consortium and processing materials. In this study, two commonly consumed traditional condiments (iru and ogiri) and their respective raw seeds (locust bean and melon) purchased from markets in south-western Nigeria were assessed for bacterial diversity and mycotoxin contamination using 16S rRNA gene sequencing and liquid chromatography tandem mass spectrometry (LC-MS/MS), respectively. Two hundred isolates obtained from the raw seeds and condiments clustered into 10 operational taxonomic units (OTUs) and spanned 3 phyla, 10 genera, 14 species and 2 sub-species. Bacillus (25%) and Staphylococcus (23.5%) dominated other genera. Potentially pathogenic species such as Alcaligenes faecalis, Bacillus anthracis, Proteus mirabilis and Staphylococcus sciuri subsp. sciuri occurred in the samples, suggesting poor hygienic practice during production and/or handling of the condiments. A total of 48 microbial metabolites including 7 mycotoxins [3-nitropropionic acid, aflatoxin B1 (AFB1), AFB2, beauvericin, citrinin, ochratoxin A and sterigmatocystin] were quantified in the food samples. Melon and ogiri had detectable aflatoxin levels whereas locust bean and iru did not; the overall mycotoxin levels in the food samples were low. There is a need to educate processors/vendors of these condiments on good hygienic and processing practices.

Bacterial Diversity and Mycotoxin Reduction During Maize Fermentation (Steeping) for Ogi Production.

Bacterial diversity and community structure of two maize varieties (white and yellow) during fermentation/steeping for ogi production, and the influence of spontaneous fermentation on mycotoxin reduction in the gruel were studied. A total of 142 bacterial isolates obtained at 24-96 h intervals were preliminarily identified by conventional microbiological methods while 60 selected isolates were clustered into 39 OTUs consisting of 15 species, 10 genera, and 3 phyla by 16S rRNA sequence analysis. Lactic acid bacteria constituted about 63% of all isolated bacteria and the genus Pediococcus dominated (white maize = 84.8%; yellow maize = 74.4%). Pediococcus acidilactici and Lactobacillus paraplantarum were found at all steeping intervals of white and yellow maize, respectively, while P. claussenii was present only at the climax stage of steeping white maize. In both maize varieties, P. pentosaceus was found at 24-72 h. Mycotoxin concentrations (µg/kg) in the unsteeped grains were: white maize (aflatoxin B1 = 0.60; citrinin = 85.8; cyclopiazonic acid = 23.5; fumonisins (B1/B2/B3) = 68.4-483; zearalenone = 3.3) and yellow maize (aflatoxins (B1/B2/M1) = 22.7-513; citrinin = 16,800; cyclopiazonic acid = 247; fumonisins (B1/B2/B3) = 252-1,586; zearalenone = 205). Mycotoxins in both maize varieties were significantly (p < 0.05) reduced across steeping periods. This study reports for the first time: (a) the association of L. paraplantarum, P. acidilactici, and P. claussenii with ogi production from maize, (b) citrinin occurrence in Nigerian maize and ogi, and