Yeasts and bacteria associated with kocho, an Ethiopian fermented food produced from enset (Ensete ventricosum).

Enset (Ensete ventricosum) is the basis of the staple food consumed by about 20% of the Ethiopian population. Kocho is one of the food products generated from enset by spontaneous fermentation of decorticated and pulverized pseudostem and corm sections. We isolated culturable microbes associated with kocho from different stages of fermentation. Twelve yeast species, six lactic acid bacteria (LABs) species and eleven species of aerobic bacteria were identified by sequencing ITS/D1D2 regions of 26S rDNA of yeasts and 16S rDNA of bacteria, respectively. More yeast species were identified in fresh (fermented for 2-5 days) kocho, compared to long-term (7-12 months) fermented kocho, while we observed an opposite trend for LABs. In fresh kocho, the most frequently isolated yeast species were Pichia exigua, Galactomyces geotrichum, and Pichia fermentans. From mid-term (3-4 months) kocho most frequently Candida cabralensis, G. geotrichum, and Candida ethanolica were isolated. In the long-term fermentations, the most frequently isolated yeast was Saturnispora silva. Lactobacillus plantarum was the most frequently isolated LAB in both fresh and mid-term kocho. In long-term fermented kocho, Acetobacter pasteurianus and L. plantarum were most frequently isolated. L. plantarum was consistently isolated from all the three stages of fermentation. Aerobic bacteria in fresh kocho were mostly gram-negative, with Raoultella planticola and Pantoea agglomerans being the most frequently isolated species. In long-term fermented kocho, mainly gram-positive, spore-forming bacteria of the genus Bacillus were found, among them also species of the Bacillus cereus group, Bacillus anthracis and Bacillus thurigiensis.

Distribution of mycotoxins produced by Penicillium spp. inoculated in apple jam and crème fraiche during chilled storage.

Estimations of consumer exposure to mycotoxins through surveillance of mycotoxins in the food trade are well described, but the exposure due to mouldy food in private homes is not known, and may result from removing visible mould on food and eating the rest. In this study, we followed the growth of Penicillium expansum on the surface of apple jam and Penicillium verrucosum on crème fraiche, as well as production and distribution of fungal metabolites throughout the sample (approx. 6 cm high divided into three equal layers), using a multianalyte method, over time (up to 28 days) and at 4, 8 and 15 °C. Growth rates and apparent lag times for P. expansum in apple jam at different temperatures were estimated by fitting to the Baranyi model. The growth rates were 1.7, 2.7 and 4.3 mm day-1 for storage at 4, 8 and 15 °C, respectively; apparent lag times decreased with increasing storage temperature and were 10.6, 7.9 and 2.6 days at corresponding temperatures. Patulin and roquefortine C were identified and quantified, among other fungal metabolites. Patulin was detected in all 2-cm layers of the apple jam at 15 °C. Concentrations in the upper two layers of the jar corresponded to exposures exceeding the health based guidance value (HBGV) for a normal serving size. Consequently, removal of the mouldy part is insufficient to avoid unhealthy exposure. In contrast to patulin, roquefortine C was also produced at 4 °C. The growth of P. verrucosum on crème fraiche was very restricted and could not be modelled. Despite the small colony (8 ±â€¯0.5 mm in diameter), ochratoxin A and citrinin were detected after 21 days at 15 °C in the top 2 cm layer (including the fungal colony), and at concentrations in a normal serving corresponding to an exposure above the HBGV established by EFSA for both mycotoxins. Questiomycin A, an antibiotic, was also produced in crème fraiche but in contrast to the two mycotoxins, was detected throughout all layers of the crème fraiche and was produced also at 4 and 8 °C. As a complement to a previous study, we also present production and the distribution of major fungal metabolites in apple jam and crème fraiche for some additional fungal strains (P. crustosum, P. roqueforti and P. verrucosum on apple jam and P. expansum on crème fraiche). A pilot study investigating the effect of inoculation size on toxin production may have implications for the best inoculum to use in experimental studies.