Taxonomic and molecular characterization of lactic acid bacteria and yeasts in nunu, a Ghanaian fermented milk product.

Produced from raw unpasteurized milk, nunu is a spontaneously fermented yoghurt-like product made in Ghana and other parts of West Africa. Despite the importance of nunu in the diet of many Africans, there is currently only limited information available on the microorganisms associated with nunu processing. With the aim of obtaining a deeper understanding of the process and as a first step towards developing starter cultures with desired technological properties for nunu production, a microbiological characterization of nunu processing in three different towns in the Upper East region of Ghana, namely Bolgatanga, Paga and Navrongo, was carried out. Lactic acid bacteria (LAB) and yeasts associated with nunu processing were isolated and identified using a combination of pheno- and genotypic methods including morphological and carbohydrate fermentation tests, (GTG)5-based rep-PCR, multiplex PCR, and 16S and 26S rRNA gene sequencing. The LAB counts during nunu processing increased from 4.5 ± 0.4 log cfu/ml at 0 h to 8.7 ± 1.8 log cfu/ml at 24 h of fermentation while yeasts counts increased from 2.8 ± 1.2 log cfu/ml at 0 h to 5.8 ± 0.5 log cfu/ml by the end of fermentation. Lactobacillus fermentum was the dominant LAB throughout the fermentations with Lactobacillus plantarum and Leuconostoc mesenteroides playing prominent roles during the first 6-8 h of fermentation as well. Less frequently isolated LAB included Lactobacillus helveticus, Enterococcus faecium, Enterococcus italicus, Weissella confusa and a putatively novel Lactococcus spp. The yeasts involved were identified as Candida parapsilosis, Candida rugosa, Candida tropicalis, Galactomyces geotrichum, Pichia kudriavzevii and Saccharomyces cerevisiae with P. kudriavzevii and S. cerevisiae being the dominant yeast species.

Identification of lactic acid bacteria isolated during traditional fura processing in Ghana.

Fura is a millet-based spontaneously fermented dumpling produced and consumed in parts of West Africa, particularly Nigeria, Burkina Faso and Ghana. From eight traditional fura production sites in northern Ghana, 862 lactic acid bacteria were isolated and identified to species level using a combination of genotypic and phenotypic methods including (GTG)(5)-based PCR fingerprinting and 16S rRNA gene sequencing, multiplex PCR by means of recA gene sequence comparison, conventional morphological characteristics and carbohydrate fermentation profiling. During millet dough fermentation, pH decreased from 5.6-6.4 to 4.1-3.7 and total lactic acid bacteria (LAB) counts increased from 4.4-5.3 to 7.9-9.2 log(10) (cfu/g). The initial stages of the fermentation were characterized by co-dominance of homo- and heterofermentative species of Pediococcus acidilactici, Weisella confusa, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus salivarius, and Lactobacillus paraplantarum whereas L. fermentum was dominating at the end of the fermentation. L. fermentum was predominant in all fermentations (p < 0.05) and a high uniformity was observed among production sites regarding the dominance of L. fermentum. L. fermentum and W. confusa were isolated in all production sites and almost at all fermentation stages indicating that they are indigenous to traditional fura processing. The other LAB bacteria species which comprised a minor proportion of the total LAB occurred occasionally and in an irregular pattern among the production sites.

Biodiversity of Saccharomyces cerevisiae isolated from a survey of pito production sites in various parts of Ghana.

Biodiversity among Saccharomyces cerevisiae predominating the spontaneous fermentation of Dagarti pito in Ghana was assessed. Two hundred and forty-nine isolates obtained from samples of dried yeast taken from commercial pito production sites in eight geographical regions of Ghana were characterized phenotypically by colony and cell morphology as well as carbohydrate assimilation profiling. Yeast populations ranged between 10(6) and 10(8) cfug(-1). Ninety-nine percent of the isolates (247) investigated showed macro-and micro morphological characteristics typical of S. cerevisiae. Of these, 72% (179) had assimilation profiles similar to S. cerevisiae while 28% (68) had assimilation profiles atypical of S. cerevisiae or any other member of the Saccharomyces sensu stricto complex. Amplification of the region spanning the two intergenic transcribed spacers (ITS) and the 5.8S ribosomal gene (ITS1-5.8S rDNA-ITS2), followed by restriction analysis, as well as determination of chromosome length polymorphism by pulsed field gel electrophoresis (PFGE) of 25 representative isolates strongly indicated that all belonged to S. cerevisiae, notwithstanding the phenotypic differences. Sequencing of the mitochondrial cytochrome-c oxidase II gene (COX 2) and the actin-encoding gene (ACT1) of four isolates, confirmed their close relatedness to S. cerevisiae, particularly to the type strain CBS1171 (98.7%), as well as other members of the Saccharomyces sensu stricto complex. Twenty isolates selected from eight geographical regions of Ghana and investigated for their technological properties, showed different patterns of growth and flocculation but otherwise similar technological characteristica. Most of the isolates produced pito having sensory attributes, which compared favourably with commercially produced pito.