Joseph D. Hounhouigan

Lactic acid bacteria and yeasts associated with gowé production from sorghum in Bénin

Aims:u2002 To identify the dominant micro‐organisms involved in the production of gowé, a fermented beverage, and to select the most appropriate species for starter culture development.

The microbiology of alkaline-fermentation of indigenous seeds used as food condiments in Africa and Asia

Alkaline-fermented food condiments play an important role in the diets of many people in developing and a few developed countries. The rise in pH during production of these foods is due to the ability of the dominant microorganisms, Bacillus spp., to hydrolyze proteins into amino acids and ammonia. Studies have been undertaken which have investigated a number of these products like dawadawa, ugba, bikalga, kinema, natto, and thua-nao. In this review, current knowledge about the principal microbiological activities and biochemical modifications which occur during the processing of the alkaline condiments including nutritional, antimicrobial, and probiotic aspects are discussed. The current use of molecular biology methods in microbiological research has allowed unambiguous and more reliable identification of microorganisms involved in these fermentations generating sufficient knowledge for the selection of potential starter cultures for controlled and better production procedures for alkaline-fermented seeds condiments.

How blanching and drying affect the colour and functional characteristics of yam (Dioscorea cayenensis-rotundata) flour

Abstract Colour and texture are important features of amala, a traditional thick paste obtained from dry yam flour. Tubers from eight yam cultivars were blanched at 65xa0°C for 20 min and dried at 40xa0°C for 5 days to test biochemical, thermal and pasting changes occurring during traditional yam flour processing. Blanching reduced peroxidase activity and drying reduced polyphenoloxidase activity, but total phenol content and the brown index of flour and of amala increased dramatically during the latter operations. The brown index of amala was significantly correlated with the total phenol content of the flour (r=0.84) and the peroxidase activity of the fresh tubers (r=0.75). Amylose content and starch gelatinization enthalpy remained stable. For all cultivars, drying significantly increased the onset gelatinization temperature, suggesting the occurrence of starch annealing. The latter leads to a reduction in swelling power and solubility during pasting, and hence to a lower paste viscosity.

The microbiota of Lafun, an African traditional cassava food product

Lafun is a fermented cassava food product consumed in parts of West Africa. In the present work the microorganisms (aerobic bacteria (AB), lactic acid bacteria (LAB) and yeasts) associated with the fermentation of Lafun under traditional conditions have for the first time been studied using a combination of pheno- and genotypic methods. During Lafun fermentation the AB count ranged from 6-7 log(10) CFU/g at the beginning to 9 log(10) CFU/g at the end. Similarly, the number of LAB increased from 5 log(10) CFU/g to 9 log(10) CFU/g during the process while the yeast load increased from 3 log(10) CFU/g at the onset of the fermentation to 5-6 log(10) CFU/g at the end of the fermentation. A total of 168 isolates (31 AB, 88 LAB, and 49 yeasts) were isolated and identified by means of phenotypic tests, PCR-based methods and 16S rRNA gene sequencing. The aerobic bacteria were mostly identified as belonging to the Bacillus cereus group (71%). The B. cereus isolates lacked the genetic determinant specific for cereulide producers but harboured several genes encoding the heat-labile toxins hemolysin BL and nonhemolytic enterotoxin as detected by PCR. The other aerobic bacteria isolated were Gram negative and identified as Klebsiella pneumoniae and Pantoea agglomerans. The dominant LAB were identified as Lactobacillus fermentum (42% of LAB isolates) followed by Lactobacillus plantarum (30%) and Weissella confusa (18%). Seven isolates remained unidentified and constitute probably a novel LAB species. The predominant yeast species associated with Lafun fermentation were Saccharomyces cerevisiae (22% of yeast isolates), Pichia scutulata (20%), Kluyveromyces marxianus (18%), Hanseniaspora guilliermondii (12%), Pichia rhodanensis (8%) and Candida glabrata (8%) as well as Pichia kudriavzevii, Candida tropicalis and Trichosporon asahii at lower incidence (<5% each).

Uncommonly High Levels of 3-Deoxyanthocyanidins and Antioxidant Capacity in the Leaf Sheaths of Dye Sorghum

Extracts from leaf sheaths of farmers varieties of dye sorghum cultivated and used in Benin as a source of biocolorings were analyzed for their anthocyanidin and phenolic contents, as well as their antioxidant capacity. The aim was to identify and quantify the types of anthocyanin and phenolic acids. The total anthocyanin content of the leaf sheaths ranged from 13.7 to 35.5 mg of cyanidin 3-glucoside equivalent/g of dry matter (DM), with an average of 27.0 mg/g. The total anthocyanin content is 90 times higher than levels usually reported in fruits and vegetables. Anthocyanin consisted essentially of apigeninidin and luteolinidin, two 3-deoxyanthocyanidins with many applications in food, beverage, and pharmaceutical industries. The apigeninidin content of the leaf sheaths was 30 times higher than that in cereal bran and ranged from 14.7 to 45.8 mg/g, with an average of 31.3 mg/g. The amount of luteolinidin ranged from 0.4 to 2.4 mg/g, with a mean of 1.2 mg/g. The total phenolic content expressed as gallic acid equivalent averaged 95.5 mg/g. The free phenolic acids identified were benzoic acid, p-coumaric acid, and o-coumaric acid at amounts of 801.4, 681.6, and 67.9 μg/g, respectively. The leaf sheaths of dye sorghum have an antioxidant capacity [3.8-5.6 mmol of Trolox equivalent (TE)/g of DM] much higher than that reported for cereal bran and fruits and vegetables.

Yeast dynamics during spontaneous fermentation of mawè and tchoukoutou, two traditional products from Benin

Mawè and tchoukoutou are two traditional fermented foods largely consumed in Benin, West Africa. Their preparations remain as a house art and they are the result of spontaneous fermentation processes. In this study, dynamics of the yeast populations occurring during spontaneous fermentations of mawè and tchoukoutou were investigated using both culture-dependent and -independent approaches. For each product, two productions were followed. Samples were taken at different fermentation times and yeasts were isolated, resulting in the collection of 177 isolates. They were identified by the PCR-DGGE technique followed by the sequencing of the D1/D2 domain of the 26S rRNA gene. The predominant yeast species identified were typed by rep-PCR. Candida krusei was the predominant yeast species in mawè fermentation followed by Candida glabrata and Kluyveromyces marxianus. Other yeast species were detected in lower numbers. The yeast successions that took place during mawè fermentation lead to a final population comprising Saccharomyces cerevisiae, C. krusei and K. marxianus. The yeast populations dominating the fermentation of tchoukoutou were found to consist of S. cerevisiae, almost exclusively. Other yeast species were detected in the early stages of fermentation. For the predominant species a succession of biotypes was demonstrated by rep-PCR for the fermentation of both products. The direct analysis at DNA and RNA levels in the case of mawè did not reveal any other species except those already identified by culture-based analysis. On the other hand, for tchoukoutou, four species were identified that were not detected by the culture-based approach. The spontaneous fermentation of mawè and tchoukoutou in the end were dominated by a few autochthonous species.

Determination of yeast diversity in ogi, mawè, gowé and tchoukoutou by using culture-dependent and -independent methods.

The maize based ogi and mawè and the sorghum based gowé and tchoukoutou are traditional, spontaneously fermented products widely consumed by the population of Benin (West Africa). Yeast occurrence in the products, as sold on local markets at different locations, was studied using a combination of culture-dependent and independent methods. Number of yeasts is varied from 3.75 log10 colony forming units (cfu)/g for ogi to 5.60 log10 cfu/g for tchoukoutou. Isolated yeasts (236) were identified based on different migration profiles on denaturing gradient gel electrophoresis (DGGE) and 26S rRNA gene sequencing. Candida krusei was the yeast most frequently isolated with strongest predominance in the maize based products. Other predominant yeast present at equal or lower incidence were Clavispora lusitaniae and Saccharomyces cerevisiae in ogi and mawè, Cl. lusitaniae, Candida tropicalis and Kluyveromyces marxianus in gowè and Cl. lusitaniae, S. cerevisiae and Candida rugosa in tchoukoutou. Grouping of C. krusei isolates (164) by rep-PCR analysis indicated that several biotypes were involved in fermentation of the four products. The DGGE analysis on the DNA directly extracted from the food matrices demonstrated the presence of Dekkera bruxellensis and Debaryomyces hansenii, not detected by the culture-based approach. This is the first study combining culture-dependent and independent methods to reveal predominant yeast species and biotypes in traditional foods from Benin.

Weissella beninensis sp. nov., a motile lactic acid bacterium from submerged cassava fermentations, and emended description of the genus Weissella

Four Gram-positive, catalase-negative, short rod-shaped or coccoid, heterofermentative lactic acid bacterial strains (2L24P13(T), 1L48P15, 1L24P31 and 1L24P34) with unusual phenotypic and genotypic properties were isolated from submerged fermenting cassava on MRS agar. All strains were motile, grew at 15 degrees C, produced dl-lactic acid from glucose with gas formation and produced ammonia from arginine. Acid was produced from d-fructose, d-galactose, d-glucose, lactose, maltose, d-mannose, melibiose, d-raffinose, sucrose, N-acetylglucosamine and d-mannitol, but not from d-arabinose or xylose. 16S rRNA gene sequence analysis revealed that the strains belonged to the genus Weissella and were most closely related to Weissella ghanensis LMG 24286(T). Low DNA-DNA reassociation values were obtained between the isolates and W. ghanensis DSM 19935(T). Based on the genetic and phenotypic results, the strains are considered to represent a novel species, for which the name Weissella beninensis sp. nov. is proposed. The type strain is 2L24P13(T) (=DSM 22752(T)=LMG 25373(T)).

Effect of storage conditions on microbiological and physicochemical quality of shea butter

Storage conditions are key constraints for quality assurance of the shea (Vitellaria paradoxa Gaertner) butter. In the Sudan savannah Africa, storage conditions of butter produced by women vary across and among processors, traders and consumers. These conditions could impact the quality of the products and reduced their access to international market. The present study attempted to investigate the effect of storage duration and packaging materials on microbiological and physicochemical characteristics of shea butter under tropical climatic conditions. Five packaging materials traditionally used in shea butter value chain were tested for their efficacy in storing shea butter freshly produced. Total germs, yeasts and mould varied with packaging materials and storage duration. After 2xa0months of storage, moisture content of butter remained constant (5%) whereas acid value increased from 3.3 to 5.4xa0mg KOH/g, peroxide value from 8.1 to 10.1xa0meq O2/kg and iodine value dropped from 48.8 to 46.2xa0mg I2/100xa0g in shea butter irrespectively to the storage materials used. The basket papered with jute bag was the less effective in ensuring the quality of butter during storage while plastic containers and plastic bags seemed to be the best packaging materials.

Physicochemical and micro-structural properties of flours, starch and proteins from two varieties of legumes: bambara groundnut (Vigna subterranea).

This work is part of a large study aimed to evaluate the potential of bambara groundnut (Vigna subterranea) flour as starting raw material for the preparation of a widely cherished legume-based food product known as koki. Towards this objective, the flours from two varieties of bambara groundnut along with their respective starch and protein isolates were analyzed for some physicochemical and microstructural properties. It was observed that bambara flour contained appreciable amount of proteins (24.0–25.5xa0g/100xa0g), carbohydrates (57.9–61.7xa0g/100xa0g), fiber (3.45–3.68xa0g/100xa0g) and ash (3.65–3.85xa0g/100xa0g) with marginal differences between both varieties. The properties of starch and proteins isolated from the flours were different from one variety to another. In particular the starch granules of the white variety were larger (size range 10–35xa0μm) and polygonal while those from the black variety were smaller (size range 6–15xa0μm) and spherical in shape. In addition, the peak of gelatinization temperature was higher for white variety (81.7xa0°C) than for black variety (77.5xa0°C). The gelatinization temperature and the enthalpy of gelatinization of starch in the flours were systematically lower than for the starch isolates, suggesting an interaction of starch with other components on the gelatinization process.