Kwaku Tano-Debrah

Yeast populations associated with Ghanaian cocoa fermentations analysed using denaturing gradient gel electrophoresis (DGGE).

The yeast populations associated with the fermentation of Ghanaian cocoa were investigated using denaturing gradient gel electrophoresis (DGGE). Samples were collected at 12–24 h intervals from heap and tray fermentations, at three different fermentation sites and different periods during the season. Eukaryotic universal primers were used to amplify a fragment of the 26S rRNA gene. The DGGE profiles were relatively complex, underlining that the fermentation of cocoa is a complex microbial process. The identities of selected fragments in the denaturing gels were revealed by sequencing. Hanseniaspora guilliermondii, Candida krusei and Pichia membranifaciens were detected from most fermentations, indicating their possible important role in the fermentation of Ghanaian cocoa. Saccharomyces cerevisiae and Candida zemplinina were almost exclusively detected during tray fermentations. The developed DGGE protocol was compared with traditional culture‐based isolations. The results were comparable but slightly different, as one yeast species (C. zemplinina) was only detected using DGGE. On the other hand, Trichosporon asahii yielded only faint bands in the denaturing gels, despite the fact that it was detected using culture‐based methods. Analysis of pure cultures showed that the targeted region of the 26S rRNA gene was poorly amplified in T. asahii, whereas all other investigated isolates were amplified efficiently using the chosen PCR approach. Cluster analysis revealed that the DGGE profiles clustered according to fermentation method and fermentation site. Furthermore, clustering according to progress in the fermentation was observed. The DGGE technique therefore seems to offer a relatively fast and reliable method for studying yeast population dynamics during cocoa fermentations. The nucleotide sequences determined in this study have been assigned Genbank Accession Nos AY762330–AY762349. Copyright

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.

Microorganisms associated with Maari, a Baobab seed fermented product

A microbiological study was carried out on Baobab fermented seeds (Maari) obtained from 4 different production sites in Burkina Faso (Mansila, Toulfé, Ouagadougou and Gorgadji). A total of 390 representative isolates comprising 251 aerobic mesophilic bacteria (AMB) and 139 lactic acid bacteria (LAB) were isolated and identified to species level using a combination of pheno- and genotypic methods including conventional morphological analysis, carbohydrate fermentation profiling, rep-PCR ((GTG)(5)-fingerprinting) and 16S rRNA gene sequencing. The fermentation of Baobab seeds was initiated by the AMB identified as Bacillus subtilis (82% of AMB isolates) and Staphylococcus sciuri (18% of AMB isolates). No lactic acid bacteria were isolated at the beginning of the process. After 24h fermentation time, Enterococcus faecium appeared in the fermenting seeds and remained until the end of the fermentation, as the predominant LAB. In Maari collected from retail outlets the AMB count ranged from 6.7log10CFU/g to 10log10CFU/g while the LAB load ranged from 4.4log10CFU/g to 9.9log10CFU/g. The AMB were identified as belonging to genus Bacillus (12 species), Staphylococcus (3 species) and one species of Aerococcus, Macrococcus, Leifsonia, Kurthia, Proteus, Acinetobacter and Globicatella, respectively. A putatively novel, previously undescribed Corynebacterium sp. was also found. E. faecium was the dominant LAB in all investigated retail samples except one sample dominated by Pediococcus acidilactici.

Microbial modification of the texture of grated cassava during fermentation into akyeke

The traditional akyeke inoculum and fermenting akyeke, an indigenous cassava product, were investigated to identify microbial species responsible for the modification of cassava texture during fermentation. Both field and laboratory samples were examined and only some cultures isolated on Plate Count Agar and Malt Extract Agar were found to be capable of causing a softening of cassava tissue when plated directly on sterile cassava slices. The cassava tissue softening isolates on PCA were tentatively identified as Bacillus subtilis and isolates on MEA as Candida tropicalis and Zygosacchromyces florentinus. The population of B. subtilis in the laboratory sample of inoculum was found to be 2.4 x 10(9) cfu g(-1) and increased during dough fermentation from 1.1 x 10(7) to 3.5 x 10(9) cfu g(-1) after 96 h. C. tropicalis was present in the inoclum at 3.0 x 10(9) cfu g(-1) and increased during dough fermentation from 3.2 x 10(6) to 6.9 x 10(7) cfu g(-1) whilst Z. florentinus was present in the inoclum at 9.1 x 10(8) cfu g(-1) and increased from 8.1 x 10(5) to 7.5 x 10(6) cfu g(-1) during dough fermentation.

Prevalence, virulence factor genes and antibiotic resistance of Bacillus cereus sensu lato isolated from dairy farms and traditional dairy products

BackgroundB. cereus are of particular interest in food safety and public health because of their capacity to cause food spoilage and disease through the production of various toxins. The aim of this study was to determine the prevalence, virulence factor genes and antibiotic resistance profile of B. cereus sensu lato isolated from cattle grazing soils and dairy products in Ghana. A total of 114 samples made up of 25 soil collected from cattle grazing farm land, 30 raw milk, 28 nunu (yoghurt-like product) and 31 woagashie (West African soft cheese). Ninety-six B. cereus sensu lato isolates from 54 positive samples were screened by PCR for the presence of 8 enterotoxigenic genes (hblA, hblC, hblD, nheA, nheB, nheC, cytK and entFM), and one emetic gene (ces). Phenotypic resistance to 15 antibiotics were also determined for 96 B. cereus sensu lato isolates.ResultsAbout 72% (18 of 25 soil), 47% (14 of 30 raw milk), 35% (10 of 28 nunu) and 39% (12 of 31 woagashi) were positive for B. cereus sensu lato with mean counts (log10 cfu/g) of 4.2 ± 1.8, 3.3 ± 2.0, 1.8 ± 1.4 and 2.6 ± 1.8 respectively. The distribution of enterotoxigenic genes revealed that 13% (12/96 isolates) harboured all three gene encoding for haemolytic enterotoxin HBL complex genes (hblA, hblC and hblD), 25% (24/96 isolates) possessed no HBL gene, whereas 63% (60/96 isolates) possessed at least one of the three HBL genes. All three genes encoding for non-haemolytic enterotoxin (nheA, nheB and nheC) were detected in 60% (57/96) isolates, 14% (13/96) harboured only one gene, 19% (18/96) whereas 8% possessed none of the NHE genes. The detection rates of cytk, entFM, and ces genes were 75, 67 and 9% respectively. Bacillus cereus s. l. isolates were generally resistant to β-lactam antibiotics such as ampicillin (98%), oxacillin (92%), penicillin (100%), amoxicillin (100%), and cefepime (100%) but susceptible to other antibiotics tested.ConclusionsBacillus cereus s. l. is prevalent in soil, raw milk and dairy products in Ghana. However, loads are at levels considered to be safe for consumption. Various enterotoxin genes associated with virulence of B. cereus are widespread among the isolates.

Improving complementary feeding in Ghana: reaching the vulnerable through innovative business--the case of KOKO Plus.

Reaching vulnerable populations in low‐resource settings with effective business solutions is critical, given the global nature of food and nutrition security. Over a third of deaths of children under 5 years of age are directly or indirectly caused by undernutrition. The Lancet series on malnutrition (2013) estimates that over 220,000 lives of children under 5 years of age can be saved through the implementation of an infant and young child feeding and care package. A unique project being undertaken in Ghana aims to bring in two elements of innovation in infant and young child feeding. The first involves a public–private partnership (PPP) to develop and test the efficacy and effectiveness of the delivery of a low‐cost complementary food supplement in Ghana called KOKO Plus™. The second involves the testing of the concepts of social entrepreneurship and social business models in the distribution and delivery of the product. This paper shares information on the ongoing activities in the testing of concepts of PPPs, social business, social marketing, and demand creation using different delivery platforms to achieve optimal nutrition in Ghanaian infants and young children in the first 2 years of life. It also focuses on outlining the concept of using PPP and base‐of‐the‐pyramid approaches toward achieving nutrition objectives.

The Use of Lactic Acid Bacteria Starter Culture in the Production of Nunu, a Spontaneously Fermented Milk Product in Ghana

Nunu, a spontaneously fermented yoghurt-like product, is produced and consumed in parts of West Africa. A total of 373 predominant lactic acid bacteria (LAB) previously isolated and identified from Nunu product were assessed in vitro for their technological properties (acidification, exopolysaccharides production, lipolysis, proteolysis and antimicrobial activities). Following the determination of technological properties, Lactobacillus fermentum 22-16, Lactobacillus plantarum 8-2, Lactobacillus helveticus 22-7, and Leuconostoc mesenteroides 14-11 were used as single and combined starter cultures for Nunu fermentation. Starter culture fermented Nunu samples were assessed for amino acids profile and rate of acidification and were subsequently evaluated for consumer acceptability. For acidification properties, 82%, 59%, 34%, and 20% of strains belonging to Lactobacillus helveticus, L. plantarum, L. fermentum, and Leu. mesenteriodes, respectively, demonstrated fast acidification properties. High proteolytic activity (>100 to 150 μg/mL) was observed for 50% Leu. mesenteroides, 40% L. fermentum, 41% L. helveticus, 27% L. plantarum, and 10% Ent. faecium species. In starter culture fermented Nunu samples, all amino acids determined were detected in Nunu fermented with single starters of L. plantarum and L. helveticus and combined starter of L. fermntum and L. helveticus. Consumer sensory analysis showed varying degrees of acceptability for Nunu fermented with the different starter cultures.

Optimisation of chocolate formulation using dehydrated peanut-cowpea milk to replace dairy milk.

BACKGROUND The rheological properties of chocolate, based upon its acceptability by consumers, are determined largely by the ingredients and their proportions used in the formulations. Milk chocolates are very popular because milk provides flavour and smooth texture to the product. This study aimed to determine the optimal ingredient formulation for vegetable milk chocolate using peanut-cowpea milk as a substitute for dairy milk. The study followed a four-component constrained mixture design, with cocoa liquor, vegetable milk, cocoa butter and sugar as the components. Lecithin and vanillin were added at a constant amount to all formulations. Critical attributes of the chocolates were evaluated using descriptive sensory tests and instrumental techniques. RESULTS Regression models were fitted to the data, and the optimum ingredient formulation for acceptable vegetable milk chocolate was determined. The vegetable milk had significant (P = 0.05) influence on flavour, mouth feel, hardness and after taste of chocolates. CONCLUSIONS The optimum ingredient formulation for acceptable vegetable milk chocolates was determined to be cocoa liquor (18.00%), sugar (30.75%), peanut-cowpea milk (28.93%), and cocoa butter (22.32%). The results demonstrate that it is feasible to use vegetable source milk for chocolate. The findings also provide clues for scale-up criteria for large-scale production of vegetable milk chocolate.

Optimization of the Nutrient Content and Protein Quality of Cereal—Legume Blends for Use as Complementary Foods in Ghana

Background Nutritionally adequate complementary foods made from locally available ingredients are of high priority in developing countries, including Ghana. The majority of complementary foods in these countries are cereal-based and are unable to meet the nutrient intakes recommended by the World Health Organization. Objective To evaluate the nutrient content and protein quality of local cereal—legume blends for complementary foods against recommendations and to determine the quantities of additional ingredients required to meet needs by using linear programming. Methods Nine cereal—legume combinations (maize, sorghum, or millet combined with cowpea, peanut, or soybean) and koko (a traditional Ghanaian maize-based complementary food) were evaluated based on the macronutrient targets for a daily ration of complementary food for the age group 12 to 24 months: 264 kcal, 6.5 g of protein, and 8.2 to 11.7 g of fat. Protein quality was assessed by the Protein Digestibility Corrected Amino Acid Score (PDCAAS). Linear programming was then used to determine the amounts of additional oil, sugar, and lysine needed to meet macronutrient requirements. Results No traditional cereal—legume food met all complementary food macronutrient requirements on its own. Cereal—legume blends made with peanut or cowpeas were low in quality protein, while those with soybean were low in fat. Lysine was the limiting amino acid (PDCAAS 0.50 to 0.82) in all blends. Adding lysine increased utilizable protein by 1% to 10% in soybean blends, 35% to 40% in peanut blends, and 14% to 24% in cowpea blends. Peanut—maize, peanut—millet, and all soybean—cereal blends were able to meet macronutrient targets; most micronutrients remained below recommended levels. Conclusions Traditional cereal—legume blends made from locally available ingredients do not meet energy, quality protein, and fat recommendations for complementary foods; however, such complementary food blends may be optimized to meet nutrient requirements by using linear programming as a tool to determine the exact levels of fortificants to be added (including, but not limited to, added fat, amino acids, and micronutrients).

Assessing Program Coverage of Two Approaches to Distributing a Complementary Feeding Supplement to Infants and Young Children in Ghana

The work reported here assesses the coverage achieved by two sales-based approaches to distributing a complementary food supplement (KOKO Plus™) to infants and young children in Ghana. Delivery Model 1 was conducted in the Northern Region of Ghana and used a mixture of health extension workers (delivering behavior change communications and demand creation activities at primary healthcare centers and in the community) and petty traders recruited from among beneficiaries of a local microfinance initiative (responsible for the sale of the complementary food supplement at market stalls and house to house). Delivery Model 2 was conducted in the Eastern Region of Ghana and used a market-based approach, with the product being sold through micro-retail routes (i.e., small shops and roadside stalls) in three districts supported by behavior change communications and demand creation activities led by a local social marketing company. Both delivery models were implemented sub-nationally as 1-year pilot programs, with the aim of informing the design of a scaled-up program. A series of cross-sectional coverage surveys was implemented in each program area. Results from these surveys show that Delivery Model 1 was successful in achieving and sustaining high (i.e., 86%) effective coverage (i.e., the child had been given the product at least once in the previous 7 days) during implementation. Effective coverage fell to 62% within 3 months of the behavior change communications and demand creation activities stopping. Delivery Model 2 was successful in raising awareness of the product (i.e., 90% message coverage), but effective coverage was low (i.e., 9.4%). Future programming efforts should use the health extension / microfinance / petty trader approach in rural settings and consider adapting this approach for use in urban and peri-urban settings. Ongoing behavior change communications and demand creation activities is likely to be essential to the continued success of such programming.