Preliminary Studies on Isolation, Bile Tolerance and Antibiogram of Potential Probiotics (Probionts) from Locally Fermented Food Products at Beach Market, Nsukka Metropolis, Enugu State, Nigeria

Aims: The study aims to isolate, evaluate bile tolerance and antibiogram studies of potential probiotics (Lactobacillus spp) from locally fermented Food Products (Akamu, Aqua Rafa® Yoghurt, Ogiri, Okpeye) and Kunu at Beach Market, Nsukka. Study Design: A ten - fold serial dilution and spread plate method using De Man, Rogosa and Sharpe (MRS) medium was adopted for isolation of potential Probionts. Place and Duration of Study: Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences, University of Nigeria Nsukka, between March - September, 2018. Methodology: Only catalase negative and Gram positive isolates characteristic of lactobacilli were used. Bile tolerance test was performed by monitoring the bacterial growth at different Bile salt concentrations (0.2%, 0.3% and 2%). The antibiogram of the isolates was assessed using the Kirby-Bauer disc diffusion method against commercial antibiotic discs of ampicillin, vancomycin, gentamycin, ciprofloxacin, methicillin and erythromycin. Results: All the 18 screened isolates were tolerant to bile salt at 0.2 % and 0.3 % with inhibition of growth at 2 % bile concentration. All isolates were observed to be resistant to methicillin (100 %) but very sensitive to gentamycin (11%) and ciprofloxacin (22%) respectively. The isolates showed intermediate resistance to other antibiotics: vancomycin (33%), erythromycin (33%) and ampicillin (44%). The decreasing pattern of resistance was thus: methicillin > ampicillin > vancomycin and erythromycin > ciprofloxacin > gentamycin. Isolates from Yoghurt (66.67%) and Ogiri (53.33%) provided most of the resistant isolates. Methicillin would provide best antagonist potential as all the isolates exhibited very high level of resistance (100 %). Conclusion: These results suggest that all the eighteen potential Lactobacillus spp strain show potential for probiotic applications and the locally fermented food products are rich sources of probionts.

Effects of lactic acid bacteria isolated from fermented mustard on lowering cholesterol

Objective: To evaluate the ability of lactic acid bacteria (LAB) strains isolated from fermented mustard to lower the cholesterol in vitro.Methods:The ability of 50 LAB strains isolated from fermented mustard on lowering cholesterol in vitro was determined by modified o-phtshalaldehyde method. The LAB isolates were analyzed for their resistance to acid and bile salt. Strains with lowering cholesterol activity, were determined adherence to Caco-2 cells. Results: Strain B0007, B0006 and B0022 assimilated more cholesterol than BCRC10474 and BCRC 17010. The isolated strains showed tolerance to pH 3.0 for 3 h despite variations in the degree of viability and bile-tolerant strains, with more than 108 CFU/mL after incubation for 24 h at 1%oxigall in MRS. In addition, strain B0007 and B0022 identified as Lactobacillus plantarum with 16S rDNA sequences were able to adhere to the Caco-2 cell lines.Conclusions:These strains B0007 and B0022 may be potential functional sources for cholesterol-lowering activities as well as adhering to Caco-2 cell lines.

Effects of lactic acid bacteria isolated from fermented mustard on lowering cholesterol

<p><b>OBJECTIVE</b>To evaluate the ability of lactic acid bacteria (LAB) strains isolated from fermented mustard to lower the cholesterol in vitro.</p><p><b>METHODS</b>The ability of 50 LAB strains isolated from fermented mustard on lowering cholesterol in vitro was determined by modified o-phtshalaldehyde method. The LAB isolates were analyzed for their resistance to acid and bile salt. Strains with lowering cholesterol activity, were determined adherence to Caco-2 cells.</p><p><b>RESULTS</b>Strain B0007, B0006 and B0022 assimilated more cholesterol than BCRC10474 and BCRC 17010. The isolated strains showed tolerance to pH 3.0 for 3 h despite variations in the degree of viability and bile-tolerant strains, with more than 10(8) CFU/mL after incubation for 24 h at 1% oxigall in MRS. In addition, strain B0007 and B0022 identified as Lactobacillus plantarum with 16S rDNA sequences were able to adhere to the Caco-2 cell lines.</p><p><b>CONCLUSIONS</b>These strains B0007 and B0022 may be potential functional sources for cholesterol-lowering activities as well as adhering to Caco-2 cell lines.</p>

Effects of lactic acid bacteria isolated from fermented mustard on lowering cholesterol

Objective: To evaluate the ability of lactic acid bacteria (LAB) strains isolated from fermented mustard to lower the cholesterol in vitro. Methods: The ability of 50 LAB strains isolated from fermented mustard on lowering cholesterol in vitro was determined by modified o-phtshalaldehyde method. The LAB isolates were analyzed for their resistance to acid and bile salt. Strains with lowering cholesterol activity, were determined adherence to Caco-2 cells. Results: Strain B0007, B0006 and B0022 assimilated more cholesterol than BCRC10474 and BCRC 17010. The isolated strains showed tolerance to pH 3.0 for 3 h despite variations in the degree of viability and bile-tolerant strains, with more than 10

Bacillus strains as feed additives: In vitro evaluation of its potential probiotic properties / Bacilos utilizados como aditivos para piensos: Evaluación in vitro de sus potenciales propiedades probióticas / Bacilos utilizados como aditivos para concentrados: Avaliação in vitro de suas propriedades probióticas

Background: much recent attention has been devoted to the genuine value of Bacillus species as multifunctional probiotic products, which produce various extracellular enzymes that enhance feed digestibility as well as many antimicrobial compounds for the purpose of improving animal performance. Objective: to describe novel, in vitro potential probiotic properties such as acid tolerance, bile tolerance, safety, and antimicrobial activity of mesophilic and psychrophilic Bacillus strains in conjunction with their extracellular enzymatic activities. Methods: four Bacillus strains (B. sp. T3, B. sp. T4, B. sp. SM2, and B. sp. JSP1) isolated from different sources were used. Strains were identified according to 16S rDNA sequences. Escherichia coli K88, E. coli O157:H7, Salmonella enteritidis KCCM 12021, Enterococcus faecalis, Listeria monocytogenes, and Staphylococcus aureus were used as indicator bacteria for the antimicrobial activity trial. Strains were activated and cultured in tryptic soy broth (pH 7.0) or broth solidified with 1.5% agar. Results: B. sp. JSP1 was fully resistant to both pH 2 and 3, whereas B. sp. SM2 showed relatively good viability at pH 3. All strains tolerated oxgall (0.3%) bile salt and were not cytotoxic to the HEK 293 human embryonic kidney cells. Three strains, except B. sp. T3, displayed differential production of extracellular enzymes including amylase, xylanase, cellulase, protease, phytase, and α-galactosidase. In particular, B. sp. SM2 inhibited six indicator pathogens: Escherichia coli K88, E. coli O157:H7, Salmonella enteritidis, Enterococcus faecalis, Listeria monocytogenes, and Staphylococcus aureus. Conclusion: the single use of B. sp. SM2 or the mixed use of the strain combined with acid or bile tolerant Bacillus strains secreting extracellular enzymes may be an alternative to antibiotics as a feed additive in farm animal production.

Antecedentes: recientemente el valor de las especies de Bacillus como productos probióticos multifuncionales ha recibido bastante atención, debido a que estos producen varias enzimas extracelulares que potencian la digestibilidad de los alimentos, así como también compuestos antimicrobianos que mejoran el desempeño del animal. Objetivo: describir y evaluar potenciales propiedades probióticas ''in vitro'' -tales como acidez, tolerancia a la bilis, seguridad y actividad antimicrobiana- de cuatro cepas de Bacilos (B. sp. T3, B. sp. T4, B. sp. SM2 y B. sp. JSP1) aisladas de diferentes fuentes, en conjunción con sus actividades enzimáticas extracelulares. Métodos: se usaron cuatro cepas de Bacillus (B. sp. T3, B. sp. T4, B. sp. SM2, and B. sp. JSP1) aisladas de diferentes fuentes. Las cepas se identificaron de acuerdo a secuencias 16S rDNA. Escherichia coli K88, E. coli O157:H7, Salmonella enteritidis KCCM 12021, Enterococcus faecalis, Listeria monocytogenes, y Staphylococcus aureus fueron empleadas como bacterias indicadoras para el ensayo de actividad antimicrobiana. Las cepas fueron activadas y cultivadas en caldo soya tripticasa (PH 7.0) o caldo solidificado con 1.5% de agar. Resultados: el B. sp. JSP1 resultó totalmente resistente tanto a pH 2 como a pH 3, mientras que el B. sp. SM2 mostró viabilidad relativamente alta a pH 3. Todas las cepas toleraron oxgall (0.3%) de sales biliares y no resultaron citotóxicas para las células humanas HEK 293 de riñón embrionario. Tres cepas, con excepción de B. sp. T3, presentaron producción diferencial de enzimas extracelulares -incluyendo amilasa, xilanasa, celulasa, proteasa, fitasa y α-galactosidasa. En particular, el B. sp. SM2 inhibió seis indicadores patógenos (Escherichia coli K88, E. coli O157: H7, Salmonella enteritidis, Enterococcus faecalis, Listeria monocytogenes y Staphylococcus aureus). Conclusiones: el uso específico de B. sp. SM2, o el uso combinado de esta cepa junto con cepas secretoras de enzimas extracelulares y tolerantes a ácidos o bilis puede ser una alternativa para reemplazar los antibióticos frecuentemente usados como aditivos en alimentación animal.

Antecedentes: o valor das espécies de Bacillus como produtos probióticos multifuncionais tem recebido bastante atenção recentemente, devido a que produzem várias enzimas extracelulares que potenciam a digestibilidade dos alimentos, como também compostos antimicrobianos que melhoram o desempenho do animal. Objetivo: descrever e avaliar as propriedades potenciais ''in vitro'' - como acidez, tolerância à bile, segurança e atividade antimicrobial- de quatro cepas de Bacilos (B. sp. T3, B. sp. T4, B. sp. SM2, and B. sp. JSP1) isoladas de diferentes fontes, em conjunto com as suas atividades enzimáticas extracelulares. Métodos: foram usadas quatro cepas de Bacillus (B. sp. T3, B. sp. T4, B. sp. SM2, and B. sp. JSP1) isoladas de diferentes fontes. As cepas foram identificadas de acordo às sequências 16S rDNA. As seguintes bactérias foram empregadas como indicadoras no teste de atividade antimicrobiana: Escherichia coli K88, E. coli O157:H7, Salmonella enteritidis KCCM 12021, Enterococcus faecalis, Listeria monocytogenes, y Staphylococcus aureus. As cepas foram ativadas e cultivadas em caldo soja tripticase (pH 7.0) ou caldo solidificado com 1.5 de Agar. Resultados: o B. sp. JSP1 foi totalmente resistente tanto no pH 2.0 como no pH 3.0, enquanto que o B. sp. SM2 mostrou viabilidade relativamente alta no pH 3.0. Todas as cepas toleraram oxgall (0.3%) de sais biliares e não foram citotóxicas para as células humanas HEK 293 de rim embrionário. Tres cepas, com exceção de B. sp. T3, apresentaram produção diferenciada de enzimas extracelulares -incluindo amilase, xilanase, celulase, protease, fitase e α-galactosidase. Particularmente, o B. sp, SM2 inibiu seis indicadores patógenos (Escherichia coli K88, E. coli O157: H7, Salmonella enteritidis, Enterococcus faecalis, Listeria monocytogenes y Staphylococcus aureus). Conclusões: O uso específico de B. sp. SM2 ou o uso combinado desta cepa junto com as cepas secretoras de enzimas extracelulares e tolerantes a ácidos ou bile, pode ser uma alternativa para substituir os antibióticos frequentemente usados como aditivos na alimentação animal.

Effect of freeze drying process on some properties of Streptococcus thermophilus isolated from dairy products

The present investigation represents the effect of freeze drying on some properties as acid and bile tolerance of Streptococcus thermophilus MTCC 1938 culture isolated from dairy products. The cell paste obtained from milk based medium was freeze dried with a pressure of 50-100 mtorr for 24h at -40ºC. Acid and bile tolerance test exhibited 3.8-4.9 and 3.2-3.8 log counts reduction after freeze drying respectively.

Prospects, Technological Aspects and Limitations of Probiotics – A Worldwide Review.

The term “probiotics” (meaning “for life” in Greek) refers products containing live microorganisms, which increase the population of friendly intestinal bacteria upon ingestion. Several new dairy product formulations containing probiotic cultures are being developed with such microbes which when consumed offer potential health benefits viz. increased resistance to infectious diseases - particularly of the intestine, decreased duration of diarrhea, reduction in blood pressure, reduction in serum cholesterol concentration and allergy, stimulation of phagocytosis by peripheral blood leucocytes, modulation of cytokine gene expression, adjuvant effects, regression of tumors, reduction in carcinogen products, increased tolerance to lactose in lactose intolerant population etc. In addition, all potential benefits could not be achieved from just one type or strain of organism. Examples of probiotic microorganisms used in foods include Lactococcus lactis, Lactobacillus sp., Streptococcus, Enterococcus, Bifidobacterium sp., Pediococcus, Propionibacteria sp. This review paper highlights the benefits, technological aspects, world scenario and limitations of probiotic foods.

Study on the Probiotic Properties of Some Lactic Acid Bacteria Strains / 微生物学通报

The acid-and bile-tolerant,the transit tolerance,the ability to adhere to human intestinal cells and the ability to assimilate cholesterol of seven lactic acid bacteria and Bifidobacteria strains were studied by an in vitro method.The abilities to adhere to CCL-187 cells were different between all strains tested(P95% when incubated in simulated gastric and small intestinal juice for 3h and 4h respectively.All strains have the ability to grow in the presence of bile,and the ability varied among the strains.The cholesterol assimilated ranged from 25% to 54%.Enterococcus faecium A30,A31 and Lactobacillus acidophilus A878,PB1,which assimilated more than 40% cholesterol from circumstances,can be good hypocholesterolemic candidate strains for functional food.