Ivan M. Mukisa

The dominant microbial community associated with fermentation of Obushera (sorghum and millet beverages) determined by culture-dependent and culture-independent methods

Obushera includes four fermented cereal beverages from Uganda namely: Obutoko, Enturire, Ekitiribita and Obuteire, whose microbial diversity has not hitherto been fully investigated. Knowledge of the microbial diversity and dynamics in these products is crucial for understanding their safety and development of appropriate starter cultures for controlled industrial processing. Culture-dependent and culture-independent techniques including denaturating gradient gel electrophoresis (DGGE) and mixed DNA sequencing of polymerase chain reaction (PCR) amplified ribosomal RNA genes were used to study the bacteria and yeast diversity of Obushera. The pH dropped from 6.0-4.6 to 3.5-4.0 within 1-2 days for Obutoko, Enturire and Obuteire whereas that of Ekitiribita decreased to 4.4 after 4 days. Counts of lactic acid bacteria (LAB) increased from 5.0 to 11.0 log cfug(-1) and yeasts increased from 3.4 to 7.1 log cfug(-1) while coliform counts decreased from 2.0 to <1 log cfug(-1) during four days of fermentation. LAB and yeast isolates were identified by rRNA gene sequence analysis. LAB isolates included: Enterococcus spp., Lactobacillus (Lb.) plantarum, Lb. fermentum, Lb. delbrueckii, Lactococcus lactis, Leuconostoc lactis, Streptococcus (S.) infantarius subsp. infantarius, Pediococcus pentosaceus and Weisella (W.) confusa. DGGE indicated predominance of S. gallolyticus, S. infantarius subsp. infantarius, Lb. fermentum, Lb. delbrueckii, W. confusa, Lb. reuteri, Fructobacillus spp., L. lactis and L. lactis. Yeast isolates included Clavispora lusitaniae, Cyberlindnera fabianii, Issatchenkia orientalis and Saccharomyces cerevisiae. DGGE indicated predominance of S. cerevisiae in Obutoko, Enturire and Obuteire and also detected Pichia spp. and I. orientalis in Obutoko. Obushera produced in the laboratory was initially dominated by Enterobacteriaceae and later by Lactococcus spp. Enterobacteriaceae and Bacillus spp. were also detected in Ekitiribita. Development of starters for Obushera may require combinations of LAB and S. cerevisiae for Obutoko, Enturire and Obuteire and LAB for Ekitiribita.

Influence of Cofermentation by Amylolytic Lactobacillus plantarum and Lactococcus lactis Strains on the Fermentation Process and Rheology of Sorghum Porridge

ABSTRACT Amylolytic lactic acid bacteria (ALAB) can potentially replace malt in reducing the viscosity of starchy porridges. However, the drawback of using ALAB is their low and delayed amylolytic activity. This necessitates searching for efficient ALAB and strategies to improve their amylolytic activity. Two ALAB, Lactobacillus plantarum MNC 21 and Lactococcus lactis MNC 24, isolated from Obushera, were used to ferment starches in MRS broth: sorghum, millet, sweet potato, and commercial soluble starch. The amylolytic activity of MNC 21 was comparable to that of the ALAB collection strain Lb. plantarum A6, while that of MNC 24 was extremely low. MNC 21, MNC 24, and their coculture were compared to A6 and sorghum malt for ability to ferment and reduce the viscosity of sorghum porridge (11.6% dry matter). ALAB and the coculture lowered the pH from 6.2 to <4.5 within 12 h, while malt as a carrier of wild starter took about 20 h. Coculturing increased lactic acid yield by 46% and 76.8% compared to the yields of MNC 21 and MNC 24 monocultures, respectively. The coculture accumulated significantly larger (P < 0.05) amounts of maltose and diacetyl than the monocultures. Sorghum malt control and the coculture hydrolyzed more starch in sorghum porridge than the monocultures. The coculture initiated changes in the rheological parameters storage modulus (G′), loss modulus (G″), phase angle (δ), and complex viscosity (η*) earlier than its constituent monocultures. The shear viscosity of sorghum porridge was reduced significantly (P < 0.05) from 1950 cP to 110 cP (malt), 281 cP (coculture), 382 cP (MNC 21), 713 cP (MNC 24), and 722 cP (A6). Coculturing strong ALAB with weak ALAB or non-ALAB can be exploited for preparation of nutrient-dense weaning foods and increasing lactic acid yield from starchy materials.

Nutritional composition, quality, and shelf stability of processed Ruspolia nitidula (edible grasshoppers)

Abstract The nutritional and commercial potential of the edible grasshopper (Ruspolia nitidula, nsenene in Luganda), a delicacy in Uganda and many East African tribes, is limited by a short shelf life and unverified nutritional value. This research established that R. nitidula is nutritious with 36–40% protein, 41–43% fat, 2.5–3.2% carbohydrate, 2.6–3.9% ash, 11.0–14.5% dietary fiber, and 900–2300 μg/100 g total carotenoids on a dry matter basis. Sautéing was the most preferred processing method resulting in grasshoppers with a notably better aroma and flavor. After 12 weeks of storage at room temperature, processed and vacuum packed, ready‐to‐eat grasshoppers maintained their edible quality with an acid value of 3.2 mg KOH/g, a total plate count of log 1.8 cfu/g, and an overall acceptability of 6.7–7.2 on a 9‐point hedonic scale. Further research is required for extending the shelf stability beyond 12 weeks and characterizing the profile of major nutrients.

Production of organic flavor compounds by dominant lactic acid bacteria and yeasts from Obushera, a traditional sorghum malt fermented beverage

Abstract Single and mixed starter cultures of lactic acid bacteria (LAB): Weissella confusa MNC20, Lactobacillus plantarum MNC21, Lactococcus lactis MNC24 and Lactobacillus fermentum MNC34 and yeasts: Issatchenkia orientalis MNC20Y and Saccharomyces cerevisiae MNC21Y were used to produce Obushera, a fermented sorghum beverage. Microbial counts, pH, sugars, organic acids, and volatile compounds in starter culture and spontaneous fermentations were monitored during 48 hrs. Maximum counts of LAB (8.4–9.4 log cfu g−1) and yeasts (7.5 ± 0.1 cfu g−1) starter cultures were attained in 6–48 hrs. Weissella confusa, Lc. lactis, and Lb. fermentum showed possible acid sensitivity while I. orientalis produced surface films. LAB starter cultures and their combinations with S. cerevisiae lowered pH from 5.83 to <4.5 (3.50–4.13) in a shorter time (12 hrs) than spontaneous fermentations (24 hrs). Lactococcus lactis and W. confusa metabolized glucose the fastest (p < .05) during the first 6 hrs. Lactobacillus fermentum, Lb. plantarum, and S. cerevisiae utilized glucose and maltose concurrently. Lactobacillus plantarum and S. cerevisiae additionally utilized fructose. S. cerevisiae metabolized sugars the fastest (p < .05) during the first 12–24 hrs. Lactobacillus plantarum and W. confusa produced the highest (p < .05) amounts of lactate (5.43 g kg−1) and diacetyl (9.5 mg kg−1), respectively. LAB also produced acetate, ethanol, acetaldehyde, acetone, and acetoin. Coculturing LAB with S. cerevisiae reduced (p < .05) lactate and diacetyl yield. Yeasts produced high amounts of acetaldehyde and methyl alcohols. Issatchenkia orientalis produced higher (p < .05) amounts of 2‐methy‐1‐propanol and 3‐methyl‐1‐butanol than S. cerevisiae. Combinations of LAB with S. cerevisiae produced a profile flavor compounds close to that of spontaneously fermented Obushera. These combinations can be adopted for controlled fermentation of Obushera and related fermented cereal products.

Effect of tamarind (Tamarindus indica L.) seed on antioxidant activity, phytocompounds, physicochemical characteristics, and sensory acceptability of enriched cookies and mango juice.

Abstract Tamarind seeds are not consumed despite their high antioxidative activity. In this study, 0–10% tamarind seed powder (TSP) was incorporated into mango juice and cookies. Total phenolics (Folin–Ciocalteu assay), antioxidant activity (2,2‐diphenyl‐1 picrylhydrazyl (DPPH) radical scavenging assay), flavonoid (aluminum chloride assay), condensed tannins content (Vanillin‐HCl assay), and consumer acceptability (n = 50) of the products were determined. TSP increased the pH and viscosity and reduced titratable acidity of juice. Incorporation of TSP increased the: total phenolic content (6.84 ± 0.21 to 88.44 ± 0.8 mg GAE/100 mL); flavonoid (4.64 ± 0.03–21.7 ± 0.36 mg CE/100 mL); condensed tannins (0.24 ± 0.01–21.81 ± 0.08 mg CE/100 mL) and total antioxidant activity (4.65 ± 0.88–21.70 ± 0.03 mg VCE/100 mL) of juice. A similar trend was observed for cookies. Maximum sensorially acceptable TSP levels were 1.5% and 6%, respectively, for juice and cookies. TSP can thus be utilized as a source of natural antioxidants in food products.

Effect of processing, packaging and storage-temperature based hurdles on the shelf stability of sautéed ready-to-eat Ruspolia nitidula

Ruspolia nitidula (an edible grasshopper) is a seasonal delicacy in Uganda and its neighbouring countries, with a postharvest shelf stability of 1-2 days. Our previous processing research increased the shelf stability of ready-to-eat R. nitidula to 12 weeks at ambient temperature. However, there is need to double the shelf-life to cover the 20-24 weeks between swarming seasons and, increase the efficiency and cost effectiveness of the processing and packaging techniques developed. This research investigated the effectiveness of 9 preservation hurdles on the microbial (total plate count), sensorial (overall acceptability) and chemical (acid value, peroxide value and thio-barbituric acid value) stability of ready-to-eat R. nitidula. Chilled storage and ambient temperature storage of the vacuum packed product increased the shelf stability of R. nitidula from 12 to 22 weeks. After 22 weeks of storage, the products had an overall acceptability of 6-7 on a 9-point hedonic scale, a total plate count of <4 log cf...

Optimization of extrusion conditions for the production of instant grain amaranth-based porridge flour

Abstract Malnutrition is one of the foremost causes of death among children below 5 years in developing countries. Development of nutrient‐dense food formulations using locally available crops has been proposed as a means to combat this menace. This study optimized the extrusion process for the production of a nutritious amaranth‐based porridge flour. Least cost formulations containing grain amaranth, groundnut, iron‐rich beans, pumpkin, orange‐fleshed sweet potato, carrot, and maize were developed and evaluated by a sensory panel (n = 30) for acceptability using the 9‐point hedonic scale. Extrusion process of the most acceptable porridge flour was optimized by response surface methodology (RSM). Barrel temperature (130–170°C) and feed moisture content (14%–20%) were the independent variables which significantly (p < .05) affected in vitro protein digestibility, vitamin A retention, total polyphenol, phytic content, and iron and zinc extractabilities. Optimization of the extrusion process improved the nutritional quality of the instant flour.

Effect of sweet potato endogenous amylase activation on in vivo energy bioavailability and acceptability of soy-enriched orange-fleshed sweet potato complementary porridges

Abstract Energy bioavailability can be influenced by food matrix factors and processing conditions or treatments. In this study, the effects of endogenous sweet potato amylase enzyme activation and slurry solids content of soy‐enriched orange‐fleshed sweet potato (OFSP) porridges on in vivo energy bioavailability (energy, weight gain, and feed efficiency ratio) and porridge acceptability were determined. Fifty‐six weanling albino rats were randomly assigned to two blocks each having eight groups of seven rats. The rats were housed in individual cages in a well‐ventilated animal house. The intervention block had rats fed on activated porridges (held at 75°C for 15 min), while rats in the control block were fed on nonactivated porridges (boiled at 90–95°C for 10 min). The rats were fed for 28 days on 50 ml of porridge per rat per day. The four groups per block were each fed on porridges with varying amounts of total solids content (10%, 15%, 20%, and 25%). Weight gain, energy bioavailability, and feed efficiency ratio were determined at the end of the feeding period. Consumer acceptability of activated and nonactivated porridges at 25% solids content was determined using a nontrained human panel (n = 40). Activation of amylases did not significantly (p > .05) affect the bioavailable energy, cumulative weight gain, and feed efficiency of the rats. Increasing slurry solids content of activated and nonactivated porridges significantly (p < .05) increased feed efficiency ratio (−14.6 ± 11.7 to 102.3 ± 2.3), weight gain (−1.4 to 5.6 g ± 1.9 g), and bioavailable energy (702.8 ± 16.2 to 1242.8 ± 12.2 kcal). Activation of amylases reduced porridge viscosity but did not significantly influence the overall acceptability. This work demonstrates the opportunity of utilizing sweet potato amylases to facilitate the preparation of complementary porridges with appropriate viscosity and increased energy density.

Application of starter cultures in the production of Enturire – a traditional sorghum-based alcoholic beverage

Abstract Enturire is an alcoholic sorghum‐ and honey‐based beverage traditionally produced by spontaneous fermentation. Its fermentation process is lengthy (5–7 days), does not guarantee product quality and safety and thus necessitates use of pure starter cultures. This study compared a modified production process of Enturire, with honey added at the start to the traditional one (honey added 3 days into the fermentation). The study also evaluated two starter culture combinations (L. plantarum MNC 21 + S. cerevisiae MNC 21 Y) and (L. plantarum MNC 21 + W. confusa MNC 20 + S. cerevisiae MNC 21Y). Microbial counts, pH, alcohol content, titratable acidity, (TA) and consumer acceptability of the Enturire were determined. Lactic acid bacteria (LAB) and yeast counts increased from 4–6 log cfu/ml to 8–9 log cfu/ml and 3–4 log cfu/ml to 5–8 log cfu/ml, respectively. Acidification of Enturire to pH <4.5 was significantly (p < .05) faster (12 hr) with starter fermentations than in the modified (18 hr) and the traditional processes (31 hr). More alcohol (9.2%–9.4%) was produced by the starters than the spontaneous fermentations (3.24%–4.38%). The modified process without starters produced a more acceptable (p < .05) product than the traditional process. The starters produced acceptable Enturire within 12 hr with pH, acidity, and alcohol content of about 3.8, 0.7%, and 2.5%–3.5%. Both starter combinations can thus be used to produce safe and acceptable Enturire in a short time.