• Journal of Dairy Science and Biotechnology
• /
• v.30 no.1
• /
• pp.45-53
• /
• 2012

Lactobacillus acidophilus isolated from the feces of a 7-day-old breast-fed infant was characterized to examine the scope of its commercial use. Forty-three Lactobacillus strains, which could grow at pH 5.5, were isolated. From these Lactobacillus isolates, 14 Lactobacillus strains were selected, which demonstrated more than 80% viability and homofermentative lactic fermentation. Finally, 9 L. acidophilus strains (NB 201~NB 209) were identified as candidate strains based upon biochemical properties, carbohydrate utilization, and cellular fatty acid composition. L. acidophilus isolates demonstrated a survival rate of more than 80% when exposed to pH 2.5 for 2 h. In particular, L. acidophilus NB 204 showed a strong acid tolerance, with a 71% survival rate even at pH 2.0. L. acidophilus isolates also manifested high bile acid tolerance; more than 87% of the cells survived on agar containing 1% bile extract, except for L. acidophilus NB 206, which showed a 73% survival rate. All L. acidophilus isolates were confirmed to have proteolytic activity; L. acidophilus NB 204 and NB 209 yielded higher levels of TCA-soluble peptides and free amino acids. The ${\beta}$-galactosidase activity of the L. acidophilus isolates was in the range of 1.97~2.45 units/mL.

• Journal of Animal Science and Technology
• /
• v.49 no.5
• /
• pp.621-632
• /
• 2007

The aim of this study was to isolate and characterize bacteriocin-producing bacteria from the intestine of duck to use as probiotics for livestock. A total of 416 strains were isolated from the small intestine and cecum of ducks and 13 isolates were finally selected after determinging inhibitory activity against pathogenic indicators by spot-on-lawn method. The selected strains were identified as Lactobacillus salivarius JWS 58, Lactobacillus plantarum JWS 1354, Pediococcus pentosaceus JWS 939, 7 strains of enterococci, and 3 strains of Escherichia coli. Lact. salivarius JWS 58, Ent. faecium JWS 833, and Ped. pentosaceus JWS 939 showed a strong inhibitory activity against Listeria monocytogenes. E. coli JWS 108 inhibited the growth of E. coli and Staphylococcus aureus. Lact. salivarius JWS 58 strain survived almost 50% in pH 2.5 phosphate buffer for 2 hr. Ped. pentosaceus JWS 939 and Lact. plantarum JWS 1354 showed strong amylolytic activity. These results suggest that a combination of bacteriocins or multispecies probiotics of the selected strains has a strong potential of alternative to antibiotics in livestock production.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.10
• /
• pp.1495-1500
• /
• 2008

In an attempt to develop a probiotic formulation for poultry feed, a number of lactic acid bacteria (LAB) were isolated from chicken intestinal specimens and a series of in vitro experiments were performed to evaluate their efficacy as a potential probiotic candidate. A total of 650 LAB strains were isolated and screened for their antagonistic potential against each other. Among all the isolates only three isolates (TMU121, 094 and 457) demonstrated a wide spectrum of inhibition and were thus selected for detailed investigations. All three selected isolates were able to inhibit the growth of E. coli and Salmonella species, although to variable extent. The nature of the inhibitory substance produced by the isolates TMU121 and 094 appeared to be associated with bacteriocin, as their activity was completely lost after treatment with proteolytic enzymes, while pH neutralization and catalase enzyme had no effect on the residual activity. In contrast, isolate TMU457 was able to resist the effect of proteolytic enzymes while pH neutralization completely destroyed its activity. Attempts were made to study the acid, bile tolerance and cell surface hydrophobicity of these isolates. TMU121 showed high bile salt tolerance (0.3%) and high cell surface hydrophobicity compared to the other two strains studied, while TMU094 appeared the most pH resistant strain. Based on these results, the three selected LAB isolates were considered as potential ingredients for a chicken probiotic feed formulation and were identified to species level based on their carbohydrate fermentation pattern by using API 50CH test kits. The three strains were identified as Lactobacillus fermentum TMU121, Lactobacillus rhamnosus TMU094, and Pediococcus pentosaceous TMU457.

• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.10
• /
• pp.1490-1499
• /
• 2016

In this study, 69 lactobacilli isolated from Tibetan Qula, a raw yak milk cheese, were screened for their potential use as probiotics. The isolates were tested in terms of: Their ability to survive at pH 2.0, pH 3.0, and in the presence of 0.3% bile salts; tolerance of simulated gastric and intestinal juices; antimicrobial activity; sensitivity against 11 specific antibiotics; and their cell surface hydrophobicity. The results show that out of the 69 strains, 29 strains (42%) had survival rates above 90% after 2 h of incubation at pH values of 2.0 or 3.0. Of these 29 strains, 21 strains showed a tolerance for 0.3% bile salt. Incubation of these 21 isolates in simulated gastrointestinal fluid for 3 h revealed survival rates above 90%; the survival rate for 20 of these isolates remained above 90% after 4 h of incubation in simulated intestinal fluid. The viable counts of bacteria after incubation in simulated gastric fluid for 3 h and simulated intestinal fluid for 4 h were both significantly different compared with the counts at 0 h (p<0.001). Further screening performed on the above 20 isolates indicated that all 20 lactobacilli strains exhibited inhibitory activity against Micrococcus luteus ATCC 4698, Bacillus subtilis ATCC 6633, Listeria monocytogenes ATCC 19115, and Salmonella enterica ATCC 43971. Moreover, all of the strains were resistant to vancomycin and streptomycin. Of the 20 strains, three were resistant to all 11 elected antibiotics (ciprofloxacin, erythromycin, tetracycline, penicillin G, ampicillin, streptomycin, polymyxin B, vancomycin, chloramphenicol, rifampicin, and gentamicin) in this study, and five were sensitive to more than half of the antibiotics. Additionally, the cell surface hydrophobicity of seven of the 20 lactobacilli strains was above 70%, including strains Lactobacillus casei 1,133 (92%), Lactobacillus plantarum 1086-1 (82%), Lactobacillus casei 1089 (81%), Lactobacillus casei 1138 (79%), Lactobacillus buchneri 1059 (78%), Lactobacillus plantarum 1141 (75%), and Lactobacillus plantarum 1197 (71%). Together, these results suggest that these seven strains are good probiotic candidates, and that tolerance against bile acid, simulated gastric and intestinal juices, antimicrobial activity, antibiotic resistance, and cell surface hydrophobicity could be adopted for preliminary screening of potentially probiotic lactobacilli.

• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.12
• /
• pp.1790-1797
• /
• 2002

Lactic acid bacteria were isolated from piglets and chicken and characterized. Lactic acid bacteria showing resistance to low pH and bile, adhesion to intestinal epithelium cells, and the inhibition of Escherichia coli and Salmonella spp. were identified as Lactobacillus acidophilus. L. acidophilus PF01 survived for 2 h in MRS broth adjusted to pH 2. L. acidophilus CF07 was less resistant than L. acidophilus PF01 to pH 2, but survived at pH 2.5 for 2 h. Both of isolates were able to grow in MRS broth containing 0.3% (w/v) bile, with L. acidophilus CF07 being more tolerant to bile than L. acidophilus PF01. L. acidophilus PF01 and CF07 adhered specifically to the duodenal and jejunal epithelium cells of piglet, and the cecal and duodenal epithelium cells of chicken, respectively. Both of isolates did not adhere to the epithelium cells of the various animal intestines from which they were isolated. When L. acidophilus was cultured with E. coli and Salmonella spp. in MRS broth, MRS broth containing 2% skim milk powder or modified tryptic soy broth at $37^{\circ}C$, L. acidophilus PF01 and CF07 inhibited the growths of E. coli K88 and K99, and S. enteritidis and S. typhimurium, respectively. Both of isolates were found to possess the essential characteristics of probiotic lactic acid bacteria for piglet and chicken.

• Asian-Australasian Journal of Animal Sciences
• /
• v.19 no.5
• /
• pp.751-755
• /
• 2006

Dadih is Indonesian traditional fermented buffalo milk believed by the natives to have beneficial effects on human health. This may be due to the probiotic properties possessed by the lactic acid bacteria (LAB) involved in its fermentation process. It was discovered that ten strains of dadih lactic isolates possessed some probiotic properties in vitro. In this study, the adhesion properties of dadih LAB, in comparison with documented probiotic strains, were investigated in vitro by using mucin extracted from human faeces and Caco-2 cells as the models for human intestinal mucosal surface and intestinal cells respectively. The adhesion results showed the distinction of Lactobacillus reuteri IS-27560 in adhering to both mucus layer and Caco-2 cells. The competition assay for adhesion to the mucus layer between dadih LAB and selected pathogens indicated the competence of Lactococcus lactis IS-16183 and Lactobacillus rhamnosus IS-7257 in significantly inhibiting the adhesion of Escherichia coli O157:H7. Accordingly, these two strains may be potential candidates for use as probiotic strains. Overall, the adhesion properties of all dadih LAB strains were relatively comparable to that of Lactobacillus casei Shirota and Lactobacillus rhamnosus GG, the documented probiotic strains.

• Microbiology and Biotechnology Letters
• /
• v.38 no.2
• /
• pp.222-226
• /
• 2010

This study was carried out to isolate and characterize the Lactobacillus plantarum with bile salt deconjugation activity that was isolated from Kimchi. Some isolates were selected and identified as L. plantarum by 16S rRNA gene sequence and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of whole cell protein patterns. They were assayed to determine their capacities to express bile salt hydrolase (BSH) activity. Among the identified strains, L. plantarum CIB 001 showed the highest level of BSH activity. Then, resistance to gastric acidity and bile condition were analyzed for further characterization. This strain was able to maintain viability for 1h at pH 2.0 and to survive in a MRS (deMan, Rogosa, and Sharpe) broth with 1.0% of bile acids. L. plantarum CIB 001 would potentially be useful in the food industry as probiotics.

• Food Science of Animal Resources
• /
• v.33 no.4
• /
• pp.522-530
• /
• 2013

The ability of probiotics to adhere to the intestinal epithelium likely plays an important role in their colonization of the gastrointestinal tract. Here, we performed high-throughput screening (HTS) for suitable characteristics of potential probiotic bacteria using attachment and colonization ability through a C. elegans surrogate in vivo model. A total of 100 strains of lactic acid bacteria (LAB) isolated from infant feces were subjected to the colonization assay using C. elegans intestine. Based on colonization ability, we showed that nine isolates have a high attachment ability during whole experimental periods (up to 168 h), compared to Lactobacillus rhamnosus strain GG as a control. Also, through the use of an in vitro cell attachment model, nine isolates revealed highly binding activity to the mucus layer. Next, the selected 9 isolates were assayed for their survival ability when exposed to acidic and bile conditions as well as cholesterol reduction and the utilization of prebiotic substrates. As a result, the isolated nine strains were determined to be highly resistant to acid and bile conditions. In addition, they have significant activity for the reduction of cholesterol and utilization of several prebiotic substrates as a carbon source. Finally, the selected nine strains were identified by either L. rhamnosus or L. plantarum (4 strains for L. rhamnosus and 5 strains for L. plantarum, respectively). Taken together, we propose that the direct colonization of probiotics using C. elegans may be applicable to the rapid screening of valuable probiotic strains in vivo.

• Food Science of Animal Resources
• /
• v.33 no.1
• /
• pp.75-82
• /
• 2013

This study aimed to isolate lactic acid bacteria (LAB) from Kimchi and to identify suitable probiotic strain for application in fermented dairy product as a commercial starter culture. A total of 106 (LAB) strains were isolated from Kimchi collected from different regions in Korea and their phenotypic characteristics were assayed. Four isolates from MRS agar plates were selected and designated as DKL109, DKL119, DKL121 and DKL128. They were identified first by API 50 CHL kit and then 16S rRNA gene sequencing. DKL121 and DKL128 were identified as Lactobacillus paracasei and Lactobacillus casei, respectively. Other two isolates (DKL109 and DKL119) were identified as Lactobacillus plantarum. To estimate their applicability in dairy products, the characteristics including acid and bile tolerance, cold shock induced cryotolerance and enzymatic activities were determined. There was wide variation in ability of strains to acid tolerance, but no significant differences in bile tolerance, cold shock induced cryotolerance within selected strains. DKL119 and DKL121 showed the highest resistance to acid and bile and the highest ${\beta}$-galactosidase activity, respectively. When these two strains were used for yogurt preparation as a single starter culture, their viable cell counts reached to $1.0{\times}10^9CFU/mL$. Lactobacillus plantarum DKL119 showed faster acid development than commercial starter culture. Also storage trials at $10^{\circ}C$ showed that the viability of these strains was retained over 15 d. With these results, it was indicated that probiotics isolated from Kimchi can be used in yogurt manufacturing as a starter culture.

• Journal of Life Science
• /
• v.22 no.2
• /
• pp.251-258
• /
• 2012

The purpose of this study was to investigate the probiotic properties of lactic acid bacterial strains isolated from animal feces. BCNU 9041 and BCNU 9042 isolates were assigned to Lactobacillus brevis on the basis of their physiological properties and 16S ribosomal DNA sequences analysis. They were confirmed as safe bioresources because of their non-hemolytic activities and non-production of harmful ${\beta}$-glucosidase, ${\beta}$-glucuronidase, tryptophanase, or urease. These isolates were also highly resistant to acid (at pH 2.5) and bile acids (at concentration of 0.3%, 0.6%, and 1% oxgall). In addition, they exhibited good antibacterial activity against food-borne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Shigella sonnei. Furthermore, it was demonstrated that they have the highest levels of hydrophobicity and that they showed bile salt hydrolytic and cholesterol assimilation activity. These results suggest that BCNU 9041 and 9042 have good potential for application in functional foods and health-related products.