Characterization of physicochemical and microbial quality, functional properties, and shelf stability of fermented tigernut-based probiotic beverages

Tigernuts and millet are indigenous underutilized crops in West Africa that have versatile applications in food processing. These crops are rich in fermentable carbohydrates, resistant starch, fiber, and micronutrients, making them ideal candidates for pre- and probiotic (synbiotic) foods. This study utilized whole tigernuts in a dairy–millet-based fermented beverage called brukina, turned it to a synbiotic, and assessed the functional and physicochemical profiles, microbial quality, and shelf stability of the beverage. The tigernut–millet agglomerate was prepared by incorporating cellulose-hydrolyzed tigernut fibrous (TNF) cake and non-hydrolyzed TNF (10% and 15%, respectively) into millet and allowing to ferment for 12 and 24 h. Brukina produced from composite tigernut milk: dairy in a ratio of 40%:60% was inoculated with the probiotic Lacticaseibacillus casei after pasteurization. The beverage was analyzed for physicochemical, proximate, and functional properties and microbiological stability at 5°C and 25°C. The obtained data were subjected to analysis of variance (ANOVA) in Minitab version 17 using a general linear model to determine the variability, interactions, and significance of the measured product characteristics. The agglomerate water absorption capacity (l/g) ranged from 0.70 ± 0.17 to 0.89 ± 0.17, bulk density (g/l) from 0.55 ± 0.04 to 0.63 ± 0.00, and swell index (%) from 1.62 ± 0.08 to 1.80 ± 0.06. The agglomerate prepared from dough and fermented for 12 h had excellent functional characteristics and was selected for synbiotic brukina production. Moisture content of the product decreased (p < 0.001) with tigernut incorporation ranging from 78.85% to 70.45%, while sodium, phosphorus, protein, total carbohydrate, and crude fiber increased with tigernut incorporation (p < 0.05). Synbiotic brukina supported the growth of L. casei attaining 11 log CFU/mL with a corresponding increase in lactic acid production and was microbiologically safe at 5°C and 25°C for 5 days compared to unpasteurized and uninoculated probiotic control (p < 0.05). The addition of whole tigernuts and L. casei to brukina enhanced its nutritional content with a shelf stability of 3 days

Prevalence and genomic characterization of Salmonella isolates from commercial chicken eggs retailed in traditional markets in Ghana

Salmonella enterica are important foodborne bacterial pathogens globally associated with poultry. Exposure to Salmonella-contaminated eggs and egg-related products is a major risk for human salmonellosis. Presently, there is a huge data gap regarding the prevalence and circulating serovars of Salmonella in chicken eggs sold in Ghana. In this study, 2,304 eggs (pools of six per sample unit) collected from informal markets in Accra, Kumasi and Tamale, representing the three ecological belts across Ghana, were tested for Salmonella. Antimicrobial susceptibility testing and Whole Genome Sequencing (WGS) of the isolates were performed using standard microdilution protocols and the Illumina NextSeq platform, respectively. The total prevalence of Salmonella was 5.5% with a higher rate of contamination in eggshell (4.9%) over egg content (1.8%). The serovars identified were S. Ajiobo (n = 1), S. Chester (n = 6), S. Hader (n = 7), S. enteritidis (n = 2); and S. I 4:b:- (n = 8). WGS analysis revealed varied sequence types (STs) that were serovar specific. The S. I 4:b:- isolates had a novel ST (ST8938), suggesting a local origin. The two S. enteritidis isolates belonged to ST11 and were identified with an invasive lineage of a global epidemic clade. All isolates were susceptible to ampicillin, azithromycin, cefotaxime, ceftazidime, gentamicin, meropenem, and tigecycline. The phenotypic resistance profiles to seven antimicrobials: chloramphenicol (13%), ciprofloxacin (94%), and nalidixic acid (94%), colistin (13%), trimethoprim (50%) sulfamethoxazole (50%) and tetracycline (50%) corresponded with the presence of antimicrobial resistance (AMR) determinants including quinolones (gyrA (D87N), qnrB81), aminoglycosides (aadA1), (aph(3“)-Ib aph(6)-Id), tetracyclines (tet(A)), phenicols (catA1), trimethoprim (dfrA14 and dfrA1). The S. enteritidis and S. Chester isolates were multidrug resistant (MDR). Several virulence factors were identified, notably cytolethal distending toxin (cdtB gene), rck, pef and spv that may promote host invasion and disease progression in humans. The findings from this study indicate the presence of multidrug resistant and virulent strains of Salmonella serovars in Ghanaian chicken eggs, with the potential to cause human infections. This is a critical baseline information that could be used for Salmonella risk assessment in the egg food chain to mitigate potential future outbreaks

Microbiological quality and antimicrobial resistance characterization of Salmonella spp. in fresh milk value chains in Ghana.

Consumer perception of poor hygiene of fresh milk products is a major barrier to promotion of milk consumption as an intervention to alleviate the burden of malnutrition in Ghana. Fresh milk is retailed raw, boiled, or processed into unfermented cheese and spontaneously fermented products in unlicensed outlets. In this study, we have determined microbiological quality of informally retailed fresh milk products and characterized the genomic diversity and antimicrobial resistance (AMR) patterns of non-typhoidal Salmonella (NTS) in implicated products. A total of 159 common dairy products were purchased from five traditional milk markets in Accra. Samples were analysed for concentrations of aerobic bacteria, total and fecal coliforms, Escherichia coli, staphylococci, lactic acid bacteria and yeast and moulds. The presence of Salmonella, E. coli O157:H7, Listeria monocytogenes and Staphylococcus aureus were determined. AMR of Salmonella against 18 antibiotics was experimentally determined. Genome sequencing of 19 Salmonella isolates allowed determination of serovars, antigenic profiles, prediction of AMR genes in silico and inference of phylogenetic relatedness between strains. Raw and heat-treated milk did not differ significantly in overall bacterial quality (P = 0.851). E. coli O157:H7 and Staphylococcus aureus were present in 34.3% and 12.9% of dairy products respectively. Multidrug resistant (MDR) Salmonella enterica serovars Muenster and Legon were identified in 11.8% and 5.9% of unfermented cheese samples respectively. Pan genome analysis revealed a total of 3712 core genes. All Salmonella strains were resistant to Trimethoprim/Sulfamethoxazole, Cefoxitin, Cefuroxime Axetil and Cefuroxime. Resistance to Chloramphenicol (18%) and Ciprofloxacin (100%), which are first line antibiotics used in treatment of NTS bacteremia in Ghana, was evident. AMR was attributed to presence and/or mutations in the following genes: golS, sdiA for cephalosporins, aac(6')-Iy, ant(9) for aminoglycosides, mdtK, gyrA, gyrB, parC, parE for quinolones and cat1, cat4 for phenicols. Phylogenetic analysis based on accessory genes clustered S. Legon strains separately from the S. Muenster strains. These strains were from different markets suggesting local circulation of related strains. Our study justifies consumer resistance to consumption of unripened soft cheese without further lethal heat treatment, and provides evidence that supports the Ghana Health Service recommendation for use of 3rd generation cephalosporins for the treatment of MDR NTS infections

Modelling the Survival of Acid-Adapted and Nonadapted Escherichia coli O157:H7 in Burkina: A Western African Traditional Fermented Milk Product

Burkina, a traditional fermented dairy product, is consumed in most parts of West Africa, including Ghana. Studies on the microflora of Burkina have indicated the presence of Escherichia coli and other pathogenic organisms. Thus, predicting the survival of E. coli in the product will inform the best manufacturing and handling practices. This study investigated the combined effect of storage temperature and time on the survival of acid-adapted and acid-non-adapted E. coli O157:H7 in Burkina. Samples were pasteurised and inoculated with acid-adapted or acid-non-adapted E. coli O157:H7. They were stored at 5, 15, and 30°C for 0, 2, 4, 6, 8, 10, 12, 14, 18, and 48 h, and the bacteria colonies were enumerated. Growth rate (survival versus time) models were developed using MATLAB software. Observed data were fitted to the Baranyi model using the DMFit curve fitting software. The E. coli O157:H7 strain appeared inherently tolerant to acid, with storage time having the most significant effect on the response parameter, survival (log CFU/mL). A negative correlation was observed for the primary models (survival versus time), which accounted for 79-97% of the relationship (p<0.05). Although E. coli survived, its growth was inhibited over time regardless of acid adaptation

DataSheet1_Characterization of physicochemical and microbial quality, functional properties, and shelf stability of fermented tigernut-based probiotic beverages.pdf

Tigernuts and millet are indigenous underutilized crops in West Africa that have versatile applications in food processing. These crops are rich in fermentable carbohydrates, resistant starch, fiber, and micronutrients, making them ideal candidates for pre- and probiotic (synbiotic) foods. This study utilized whole tigernuts in a dairy–millet-based fermented beverage called brukina, turned it to a synbiotic, and assessed the functional and physicochemical profiles, microbial quality, and shelf stability of the beverage. The tigernut–millet agglomerate was prepared by incorporating cellulose-hydrolyzed tigernut fibrous (TNF) cake and non-hydrolyzed TNF (10% and 15%, respectively) into millet and allowing to ferment for 12 and 24 h. Brukina produced from composite tigernut milk: dairy in a ratio of 40%:60% was inoculated with the probiotic Lacticaseibacillus casei after pasteurization. The beverage was analyzed for physicochemical, proximate, and functional properties and microbiological stability at 5°C and 25°C. The obtained data were subjected to analysis of variance (ANOVA) in Minitab version 17 using a general linear model to determine the variability, interactions, and significance of the measured product characteristics. The agglomerate water absorption capacity (l/g) ranged from 0.70 ± 0.17 to 0.89 ± 0.17, bulk density (g/l) from 0.55 ± 0.04 to 0.63 ± 0.00, and swell index (%) from 1.62 ± 0.08 to 1.80 ± 0.06. The agglomerate prepared from dough and fermented for 12 h had excellent functional characteristics and was selected for synbiotic brukina production. Moisture content of the product decreased (p < 0.001) with tigernut incorporation ranging from 78.85% to 70.45%, while sodium, phosphorus, protein, total carbohydrate, and crude fiber increased with tigernut incorporation (p < 0.05). Synbiotic brukina supported the growth of L. casei attaining 11 log CFU/mL with a corresponding increase in lactic acid production and was microbiologically safe at 5°C and 25°C for 5 days compared to unpasteurized and uninoculated probiotic control (p < 0.05). The addition of whole tigernuts and L. casei to brukina enhanced its nutritional content with a shelf stability of 3 days.</p

Microbial Diversity and Metabolite Profile of Fermenting Millet in the Production of <i>Hausa koko</i>, a Ghanaian Fermented Cereal Porridge

Hausa koko is an indigenous porridge processed from millet in Ghana. The process involves fermentation stages, giving the characteristic organoleptic properties of the product that is produced largely at a small-scale household level and sold as a street food. Like many other indigenous foods, quality control is problematic and depends on the skills of the processor. In order to improve the quality of the product and standardize the process for large-scale production, we need a deeper understanding of the microbial processes. The aim of this study is to investigate the microbial community involved in the production of this traditional millet porridge and the metabolites produced during processing. High-throughput amplicon sequencing was used to identify the bacterial (16S rRNA V4 hypervariable region) and fungal [Intergenic Transcribed Spacer (ITS)] communities associated with the fermentation, while nuclear magnetic resonance (NMR) was used for metabolite profiling. The bacterial community diversity was reduced during the fermentation processes with an increase and predominance of lactobacilli. Other dominant bacteria in the fermentation included Pediococcus, Weissella, Lactococcus, Streptococcus, Leuconostoc, and Acetobacter. The species Limosilactobacillus fermentum and Ligilactobacillus salivarius accounted for some of the diversities within and between fermentation time points and processors. The fungal community was dominated by the genus Saccharomyces. Other genera such as Pichia, Candida, Kluyveromyces, Nakaseomyces, Torulaspora, and Cyberlindnera were also classified. The species Saccharomyces cerevisiae, Stachybotrys sansevieriae, Malassezia restricta, Cyberlindnera fabianii, and Kluyveromyces marxianus accounted for some of the diversities within some fermentation time points. The species S. sansevieria and M. restricta may have been reported for the first time in cereal fermentation. This is the most diverse microbial community reported in Hausa koko. In this study, we could identify and quantify 33 key different metabolites produced by the interactions of the microbial communities with the millet, composed of organic compounds, sugars, amino acids and intermediary compounds, and other key fermentation compounds. An increase in the concentration of organic acids in parallel with the reduction of sugars occurred during the fermentation process while an initial increase of amino acids followed by a decrease in later fermentation steps was observed