Paulin Azokpota

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.

Identification, genetic diversity and cereulide producing ability of Bacillus cereus group strains isolated from Beninese traditional fermented food condiments.

Bacillus cereus sensu lato is often detected in spontaneously fermented African foods but is rarely identified to species level. Only some of the B. cereus group species are reported to be pathogenic to humans and identification to species level is necessary to estimate the safety of these products. In the present study, a total of 19 Bacillus cereus group spp. isolated from afitin, iru and sonru, three spontaneously fermented African locust (Parkia biglobosa) bean based condiments produced in Benin, were investigated. The strains were isolated at 6, 12, 18, 24 and 48 h fermentation time. By using phenotypic and genotypic methods all of the isolates could be identified as B. cereus sensu stricto. The isolates were grouped according to their PM13 PCR (random amplification of polymorphic DNA PCR) fingerprint and formed two major clusters, one of which contained eight strains isolated from afitin (cluster 1). Highly similar PM13 profiles were obtained for seven of the isolates, one from afitin, one from iru and five from sonru (cluster 2). Four of the isolates, one from afitin and three from sonru, did not form any particular cluster. The PM13 profiles of cluster 2 isolates were identical to those which are specific to emetic toxin producers. Cereulide production of these isolates was confirmed by liquid chromatography mass spectrometry/mass spectrometry. This is the first report on cereulide producing B. cereus in African fermented foods. Occurrence of the opportunistic human pathogen B. cereus, which is able to produce emetic toxin in afitin, iru and sonru, could impose a health hazard. Interestingly, no reports on food poisoning from the consumption of the fermented condiments exist.

Biodiversity of aerobic endospore-forming bacterial species occurring in Yanyanku and Ikpiru, fermented seeds of Hibiscus sabdariffa used to produce food condiments in Benin

Yanyanku and Ikpiru made by the fermentation of Malcavene bean (Hibiscus sabdariffa) are used as functional additives for Parkia biglobosa seed fermentations in Benin. A total of 355 aerobic endospore-forming bacteria (AEFB) isolated from Yanyanku and Ikpiru produced in northern and southern Benin were identified using phenotypic and genotypic methods, including GTG5-PCR, M13-PCR, 16S rRNA, gyrA and gyrB gene sequencing. Generally, the same 5-6 species of the genus Bacillus predominated: Bacillus subtilis (17-41% of isolates), Bacillus cereus (8-39%), Bacillus amyloliquefaciens (9-22%), Bacillus licheniformis (3-26%), Bacillus safensis (8-19%) and Bacillus altitudinis (0-19%). Bacillus aryabhattai, Bacillus flexus, and Bacillus circulans (0-2%), and species of the genera Lysinibacillus (0-14%), Paenibacillus (0-13%), Brevibacillus (0-4%), and Aneurinibacillus (0-3%) occurred sporadically. The diarrheal toxin encoding genes cytK-1, cytK-2, hblA, hblC, and hblD were present in 0%, 91% 15%, 34% and 35% of B. cereus isolates, respectively. 9% of them harbored the emetic toxin genetic determinant, cesB. This study is the first to identify the AEFB of Yanyanku and Ikpiru to species level and perform a safety evaluation based on toxin gene detections. We further suggest, that the gyrA gene can be used for differentiating the closely related species Bacillus pumilus and B. safensis.

Volatile compounds profile and sensory evaluation of Beninese condiments produced by inocula of Bacillus subtilis

BACKGROUND Three Beninese food condiments (ABS1(24h), IBS2(48h) and SBS3(48h)) were produced by controlled fermentation of African locust beans using inocula of pure cultures of Bacillus subtilis, BS1, BS2 and BS3, respectively. Quantitative and qualitative assessments of the volatile compounds in the condiments produced have been performed using the Likens-Nickerson simultaneous distillation-extraction method and GC-MS analysis, followed by a sensory evaluation in comparison with the spontaneously fermented condiments. RESULTS A total of 94 volatile compounds have been found including 53 compounds identified in relatively high concentrations and were subdivided into seven main groups with the predominance of four major groups: pyrazines, aldehydes, ketones and alcohols. Compared to the spontaneously fermented condiments, volatile compounds identified in controlled fermented condiments have been found in high number and in concentrations which varied according to the inoculum of B. subtilis used. The condiments produced with starter cultures scored significantly (P < 0.05) higher for odour than the spontaneously fermented condiments. But the overall acceptability (7/10) of the two types of condiments was similar. CONCLUSION The investigated B. subtilis, BS1, BS2 and BS3 can be considered as potential starter cultures for the fermentation of African locust beans to produce good quality of Beninese food condiments.

Prediction of spoilage of tropical shrimp (Penaeus notialis) under dynamic temperature regimes

The spoilage activity of Pseudomonas psychrophila and Carnobacterium maltaromaticum, two tropical shrimp (Penaeus notialis) spoilage organisms, was assessed in cooked shrimps stored at 0 to 28 °C. Microbiological, chemical and sensory analyses were performed during storage. P. psychrophila had a higher growth rate and showed a higher spoilage activity at temperatures from 0 to 15 °C, while at 28 °C, C. maltaromaticum had a higher growth rate. The spoilage activity of P. psychrophila was found to be higher in cooked shrimp than in fresh shrimp. Observed shelf-life data of shrimps stored at constant temperatures were used to validate a previously developed model that predicts tropical shrimp shelf-life at constant storage temperatures. Models predicting the growth of the spoilage organisms as a function of temperature were constructed. The validation of these models under dynamic storage temperatures simulating temperature fluctuation in the shrimp supply chain showed that they can be used to predict the shelf-life of cooked and fresh tropical shrimps.

Bacterial concentration and diversity in fresh tropical shrimps (Penaeus notialis) and the surrounding brackish waters and sediment.

This study aimed at determining bacterial concentration and diversity in fresh tropical shrimps (Penaeus notialis) and their surrounding brackish waters and sediment. Freshly caught shrimp, water and sediment samples were collected in Lakes Nokoue and Aheme in Benin (West Africa) during two periods with different water salinity and temperature. We used complementary culture-dependent and culture-independent methods for microbiota analysis. During both sampling periods, total mesophilic aerobic counts in shrimp samples ranged between 4.4 and 5.9 log CFU/g and were significantly higher than in water or sediment samples. In contrast, bacterial diversity was higher in sediment or water than in shrimps. The dominant phyla were Firmicutes and Proteobacteria in shrimps, Firmicutes, Proteobacteria, and Actinobacteria in water, and Proteobacteria and Chloroflexi in sediment. At species level, distinct bacterial communities were associated with sediment, water and shrimps sampled at the same site the same day. The study suggests that the bacterial community of tropical brackish water shrimps cannot be predicted from the microbiota of their aquatic environment. Thus, monitoring of microbiological quality of aquatic environments might not reflect shrimp microbiological quality.

Profiling the quality characteristics of the butter of Pentadesma butyracea with reference to shea butter.

BACKGROUND Pentadesma butyracea is a tropical plant species. Its kernels are rich in edible butter similar to shea butter. This study evaluated the quality characteristics of the Pentadesma butter produced by cottage enterprises in Benin, using a quantitative survey approach and physicochemical and sensorial analysis methods. RESULTS The butter of Pentadesma is mostly used for food preparation, cosmetic and therapeutic applications. It is characterized by a yellow colour, a hard texture, a relatively sweet taste and a bright appearance. Consumers preferred Pentadesma butter to shea butter for colour, taste, texture and appearance. Instrumental analysis showed that the average water content (6.5 g kg(-1) ) and peroxide value (0.74 mEq O2 kg(-1) ) of Pentadesma butter were lower than those of shea butter, for which average water content and peroxide values were 20.7g kg(-1) and 2.09 mEq O2 kg(-1) respectively. The fatty acid profile of Pentadesma butter is similar to that of shea butter. CONCLUSION This study showed that the Pentadesma butyracea butter produced by cottage enterprises in Benin exhibited quality characteristics which are better than that of shea butter. The data generated can be used for a better exploitation of the butter in food and cosmetic industries.

Effect of fuel and kiln type on the polycyclic aromatic hydrocarbon (PAH) levels in smoked shrimp, a Beninese food condiment

Smoked shrimp is a food condiment widely used in Beninese local cooking practices. A previous study revealed that this product is highly contaminated with polycyclic aromatic hydrocarbons (PAH). The present study explored possibilities to reduce PAH levels in shrimp smoked using cottage industry smoking techniques with barrel and chorkor kilns, by replacing wood by charcoal from Acacia auriculiformis and Mangifera indica, as fuels. Results showed that only shrimp smoked using acacia charcoal in a chorkor kiln had PAH levels (benzo[a]pyrene = 5 µg kg−1 and sum of benzo[a]pyrene, chrysene, benzo[a]anthracene and benzo[b]fluoranthene = 28 µg kg−1) in accordance with a European standard of 5 and 30 µg kg−1, respectively, and suitable physicochemical characteristics for good storage (moisture content = 11.9% ± 1.5%; water activity = 0.46 ± 0.03). However, further investigations still needs to be done to reduce the duration of product contact with combustion gasses in order to reduce the PAH content of smoked shrimp to safer levels, largely below standards.

Indigenous Perception and Characterization of Yanyanku and Ikpiru: Two Functional Additives for the Fermentation of African Locust Bean

Indigenous perception, processing methods, and physicochemical and microbiological characteristics of Yanyanku and Ikpiru, two additives used to produce fermented African locust bean condiments, Sonru and Iru, were evaluated. According to producers, these additives accelerate the fermentation and soften the texture of the condiments. Yanyanku is produced by spontaneous fermentation with either Hibiscus sabdariffa or Gossypium hirsutum or Adansonia digitata seeds, whereas only Hibiscus sabdariffa seeds are used for Ikpiru. Both additives, with pH values ranging between 6.2 and 10 and Bacillus spores varying between 5.5 and 8.9 Log10 (CFU/g), could be considered as softening additives or enrichment inocula to produce condiments.

Variations in the traditional processing methods of Pentadesma butracea butter in northern Benin

Pentadesma butyracea is used in the North of Benin to produce a type of butter similar to shea butter. The present work was carried out to investigate the traditional processing of butter from P. butyracea through a survey among 192 traditional butter processors from 14 sociocultural groups in five different parklands in northern Benin. Three different pre-treatments of P. butyracea kernels have been identified as the basis of the butter extraction process. The pre-treatment more frequently used (69.7 per cent of processors) consists of depulping the fruit to get fresh kernels which are first boiled for 1–2 h before sun drying for 14–30 days to get cooked and dried kernels which are involved in the next steps of the extraction process. The extraction itself involves two main steps, heating and kneading, which also vary according to processors. Ten methods of processing of P. butyracea including six new methods have been reported. Two of the newly identified methods are mostly used by Boo, Anii, and Mokole...