Molecular and biochemical characterizations of human oral lactobacilli as putative probiotic candidates
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Introduction: The objective of this study was to characterize the lactobacilli from the human oral cavity as a potential source of probiotic strains. Methods: Samples were collected from four different locations within the oral cavity: surface of healthy tooth, oral mucous membrane, surface of tooth decay and deep tooth decay. On the basis of morphological and biochemical properties eight categories were formed and 26 isolates were selected for further characterization. The isolates were determined as Lactobacillus sp. using primers specific for 16S rDNA. Sequencing of 16S rDNA genes and repetitive sequence‐based polymerase chain reactions were used for determination to species and subspecies levels. Results: Predominant species were Lactobacillus fermentum , Lactobacillus plantarum , Lactobacillus salivarius and Lactobacillus paracasei subsp. paracasei , while Lactobacillus acidophilus , Lactobacillus cellobiosus , Lactobacillus delbrueckii subsp. lactis and Lactobacillus gasseri were also present. The isolates Lactobacillus salivarius BGHO1, Lactobacillus fermentum BGHO36 and BGHO64, Lactobacillus gasseri BGHO89 and Lactobacillus delbrueckii subsp. lactis BGHO99 exhibited antagonistic action on the growth of Staphylococcus aureus , Enterococcus faecalis , Micrococcus flavus , Salmonella enteritidis , Streptococcus pneumoniae and Streptococcus mutans , but not on growth of Candida albicans. Moreover, the isolates L. salivarius BGHO1 and L. gasseri BGHO89 were tolerant to low pH and high concentration of bile salts. Conclusion: Taken together, these findings imply that L. salivarius BGHO1 and L. gasseri BGHO89 might be subjects for additional investigation as potential probiotic strains.Keywords:
Lactobacillus salivarius
Lactobacillus gasseri
Lactobacillus fermentum
Lactobacillus paracasei
Lactobacillus crispatus
Enterococcus faecalis
The efficacy of a biochemical kit, API50CHL kit, for identification of intestinal and vaginal lactobacilli from humans was evaluated by comparing with the results of DNA-DNA hybridization assay. The results showed that in total only 52 of the 172 strains (30.2%) tested were identified correctly by the kit at species level. Especially all strains of some species, such as Lactobacillus gasseri and Lactobacillus crispatus, were misidentified as Lactobacillus acidophilus by the kit. However, the kit was found to be reliable for identification of Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus salivarius. This suggests that the exact identification of Lactobacillus isolates from human stool and vaginal specimens by API50CHL kit is difficult without the support of modern genotypic technique.
Lactobacillus gasseri
Lactobacillus salivarius
Lactobacillus paracasei
Lactobacillus crispatus
Lactobacillus rhamnosus
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Objective Identifying the dominant Lactobacillus producing bacteriocin,which were screened from 60 healthy women vaginas,provides choice and credible strains for the research of probiotic.Methods 19 strains of Lactobacillus which produce bacteriocin and abundant lactic acid were screened by the method of Oxford-cup tests,and they were identified by several important biochemical characteristics.Results According to the results of these experiments,19 strains were identified that 9 were Lactobacillus.gasseri,1 was Lactobacillus.salivarius and 9 were Lactobacillus.crispatus.Conclusions These 19 strains Lactobacillus which were screened through this experiment were dominant beneficial bacteria in the healthy female vagina,which had high ability of producing acid and bacteriocin.16 strains of them produce H_2O_2 and some had great probiotic production value.
Lactobacillus gasseri
Lactobacillus crispatus
Lactobacillus salivarius
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About 16,000 isolates from animal feces and intestines were assayed for the production of acid urease, and the 700 strains selected as producers were examined as to their taxonomic properties. Of these 700 strains, 370 belonged to the genus Streptococcus, 312 to the genus Lactobacillus, 9 to the genus Escherichia, 6 to the genus Staphylococcus, 2 to the genus Morganella and 1 to the genus Bifidobacterium. The majority of the streptococci were identified as Streptococcus mitior, the remainder being Streptococcus salivarius, Streptococcus faecalis, Streptococcus faecium, Streptococcus avium and Streptococcus gallinarum. The majority of the lactobacilli were considered to be Lactobacillus reuteri or Lactobacillus fermentum and Lactobacillus animalis or Lactobacillus salivarius, the remainder being Lactobacillus ruminis, Lactobacillus viridescens, Lactobacillus vac- cinostercus and strains considered to be Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus crispatus or Lactobacillus gasseri.
Lactobacillus salivarius
Lactobacillus fermentum
Lactobacillus crispatus
Lactobacillus gasseri
Lactobacillus reuteri
Streptococcus salivarius
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This study aimed to identify oral Lactobacillus species and characterize their adhesion properties and antibacterial activity in patients with periodontitis compared with periodontally healthy individuals.Three hundred and fifty-four isolates from the saliva, subgingival, and tongue plaque of 59 periodontitis patients and 59 healthy individuals were analyzed. Oral Lactobacillus species were identified through the culture method in the modified MRS medium and confirmed by molecular testing. Moreover, the radial diffusion assay and cell culture methods were used to determine the antibacterial activities of oral strains against oral pathogens and their adhesion activity in vitro.67.7% of the cases and 75.7% of the control samples were positive for the Lactobacillus species. Lacticaseibacillus paracasei and Limosilactobacillus fermentum were the dominant species in the case group, whereas Lacticaseibacillus casei and Lactiplantibacillus plantarum were dominant in the control group. Lactobacillus crispatus and Lactobacillus gasseri had higher antibacterial effects against oral pathogens. Moreover, Ligilactobacillus salivarius and L. fermentum demonstrated the highest ability to adhere to oral mucosal cells and salivary-coated hydroxyapatite.L. crispatus, L. gasseri, L. fermentum, and L. salivarius can be introduced as probiotic candidates since they demonstrated appropriate adherence to oral mucosal cells and salivary-coated hydroxyapatite and also antibacterial activities. However, further studies should be conducted to assess the safety of probiotic interventions using these strains in patients with periodontal disease.
Lactobacillus fermentum
Lactobacillus salivarius
Lactobacillus gasseri
Lactobacillus casei
Lactobacillus paracasei
Lactobacillus crispatus
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Introduction: The objective of this study was to characterize the lactobacilli from the human oral cavity as a potential source of probiotic strains. Methods: Samples were collected from four different locations within the oral cavity: surface of healthy tooth, oral mucous membrane, surface of tooth decay and deep tooth decay. On the basis of morphological and biochemical properties eight categories were formed and 26 isolates were selected for further characterization. The isolates were determined as Lactobacillus sp. using primers specific for 16S rDNA. Sequencing of 16S rDNA genes and repetitive sequence‐based polymerase chain reactions were used for determination to species and subspecies levels. Results: Predominant species were Lactobacillus fermentum , Lactobacillus plantarum , Lactobacillus salivarius and Lactobacillus paracasei subsp. paracasei , while Lactobacillus acidophilus , Lactobacillus cellobiosus , Lactobacillus delbrueckii subsp. lactis and Lactobacillus gasseri were also present. The isolates Lactobacillus salivarius BGHO1, Lactobacillus fermentum BGHO36 and BGHO64, Lactobacillus gasseri BGHO89 and Lactobacillus delbrueckii subsp. lactis BGHO99 exhibited antagonistic action on the growth of Staphylococcus aureus , Enterococcus faecalis , Micrococcus flavus , Salmonella enteritidis , Streptococcus pneumoniae and Streptococcus mutans , but not on growth of Candida albicans. Moreover, the isolates L. salivarius BGHO1 and L. gasseri BGHO89 were tolerant to low pH and high concentration of bile salts. Conclusion: Taken together, these findings imply that L. salivarius BGHO1 and L. gasseri BGHO89 might be subjects for additional investigation as potential probiotic strains.
Lactobacillus salivarius
Lactobacillus gasseri
Lactobacillus fermentum
Lactobacillus paracasei
Lactobacillus crispatus
Enterococcus faecalis
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Abstract Aim: To analyze the acid‐producing and growth abilities of different oral Lactobacillus species. Methods: Thirty‐nine oral clinical strains and type strains of Lactobacillus , representing nine species, including Lactobacillus casei/paracasei , Lactobacillus fermentum , Lactobacillus gasseri , Lactobacillus mucosae , Lactobacillus oris , Lactobacillus plantarum , Lactobacillus rhamnosus , Lactobacillus salivarius , and Lactobacillus vaginalis were studied. Anaerobically‐grown bacterial cells were inoculated overnight in de Man, Rogosa, and Sharpe broth containing 2% glucose (pH 7.0). Acid production and growth were measured at 0, 1.5, 3, 5, 7, and 24 h. Results: Lactobacillus salivarius , Lactobacillus rhamnosus , Lactobacillus casei/paracasei , and Lactobacillus plantarum grew rapidly and reached an optical density higher than other species. They also produced more acid than the others. Lactobacillus vaginalis showed the lowest rate of growth and acid production. These findings demonstrated that the different species of Lactobacillus showed different abilities to generate acid, allowing the species to be categorized into three groups: strongly, moderately, and weakly acidogenic. Conclusion: There was variation in acid production and growth between the Lactobacillus species. The strongest acid producers were Lactobacillus salivarius , Lactobacillus rhamnosus , Lactobacillus plantarum , and Lactobacillus casei/paracasei , respectively. It seems possible that these species might play a more important role in caries development than the others.
Lactobacillus casei
Lactobacillus salivarius
Lactobacillus rhamnosus
Lactobacillus fermentum
Lactobacillus paracasei
Lactobacillus gasseri
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About 16, 000 isolates from animal feces and intestines were assayed for the production of acid urease, and the 700 strains selected as producers were examined as to their taxonomic properties. Of these 700 strains, 370 belonged to the genus Streptococcus, 312 to the genus Lactobacillus, 9 to the genus Escherichia, 6 to the genus Staphylococcus, 2 to the genus Morganella and 1 to the genus Bifidobacterium. The majority of the streptococci were identified as Streptococcus mitior, the remainder being Streptococcus salivarius, Streptococcus faecalis, Streptococcus faecium, Streptococcus avium and Streptococcus gallinarum. The majority of the lactobacilli were considered to be Lactobacillus reuteri or Lactobacillus fermentum and Lactobacillus animalis or Lactobacillus salivarius, the remainder being Lactobacillus ruminis, Lactobacillus viridescens, Lactobacillus vaccinostercus and strains considered to be Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus crispatus or Lactobacillus gasseri.
Lactobacillus salivarius
Lactobacillus fermentum
Lactobacillus gasseri
Lactobacillus crispatus
Lactobacillus reuteri
Streptococcus salivarius
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Objective:Isolating lactobacillus from health female genital tract, for exploring probiotics to research compound preparation.Methods:Traditional morphology analysis was used to identify lactobacillus, and further screen hydrogen peroxide-producing lactobacillus.The molecular biology method was used to identify exactly and to examine predominant strain by performance test.Results:There were 11 strains of lactobacillus isolated and identified from 18 health examination women genital tract, the detection rate was 61.1%.According to the results of these experiment,11 strains were identified that 4 were lactobacillus acidophilus, 3 were lactobacillus gasseri, 2 were lactobacillus jensenii,1 was lactobacillus crispatus and another one was lactobacillus salivarius.Conclusion:Traditional biochemical identification and molecular biology methods could identify lactobacillus.Only lactobacillus crispatus is hydrogen peroxide-producing one. After the preliminary test, the strain is a strong acid-resistant and hot-resistant one, and is bacteriostasis in vitro testing.
Lactobacillus crispatus
Lactobacillus gasseri
Lactobacillus salivarius
Isolation
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There is limited information concerning the adhesion and aggregation of human oral lactobacilli. In this study, the adhesion of 10 Lactobacillus species was investigated using H357 oral keratinocyte cells as an in vitro model for oral mucosa. Coaggregation with the representative oral pathogen, Streptococcus mutans ATCC 25175, and the physicochemical cell properties was also evaluated. The results demonstrated significant variations in adhesion (42–96%) and aggregation (autoaggregation, 14–95%; coaggregation, 19–65%). All strains showed a high affinity for chloroform, and most strains had a moderate‐to‐high hydrophobicity. All strains, except Lactobacillus casei and Lactobacillus gasseri, showed a moderate affinity for ethyl acetate. There was a strong association of autoaggregation with coaggregation (rs = 0·883, P <0·001). The highest mean for autoaggregation (74%) and coaggregation (47%) belonged to the Lact. gasseri strains. Correlations between the adhesion and surface characteristics and aggregation were observed among the Lactobacillus fermentum and Lactobacillus paracasei strains; however, there was a variation in the strains properties within and between species. This study indicated that the Lact. gasseri, Lact. fermentum, and Lact. paracasei strains might be potential probiotics for the human oral cavity given their desirable properties. It should also be emphasized that a selective process for probiotic strains is required. Adhesion to host tissues and bacterial aggregation (auto‐ and coaggregation) are the highly important criteria for selecting strains with probiotic potential. These abilities are commonly involved with surface‐charged characteristics. This is the first study to investigate the oral Lactobacillus species using an oral keratinocyte cell line. Significant results were found for the correlations between the adhesion and surface charge characteristics and for aggregation among certain strains of Lactobacillus gasseri, Lactobacillus fermentum and Lactobacillus paracasei. This observation could be useful when collecting background information for the selection of probiotic strains for use in oral health.
Lactobacillus fermentum
Lactobacillus gasseri
Lactobacillus paracasei
Lactobacillus casei
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The objective of this study was to investigate the cultivable Lactobacillus population in adult and infant faecal material to identify strains shared across a number of individuals. A range of lactobacilli isolated on Lactobacillus-selective agar from faeces of 16 infants and 11 adults were genetically fingerprinted and further characterized by 16S rRNA gene sequencing. The relatedness of all the Lactobacillus strains isolated to known species was also determined both genetically and phenotypically. This study revealed that the human intestine is initially colonized by only a few (1-2) different cultivable strains whereas in adults the pattern becomes more complex with a higher diversity of strains. The adult samples contained three genetically distinct Lactobacillus strains in some cases, while infant samples generally harboured only one dominant Lactobacillus strain. Moreover, the species in general appeared to differ with Lactobacillus rhamnosus and Lactobacillus casei/paracasei found mainly in adults, whereas Lactobacillus gasseri and Lactobacillus salivarius were more commonly isolated in infant samples. The data reaffirm the differences in Lactobacillus populations both between individual subjects and between the infant and adult, with an overall change in the diversity and complexity from early stages of life to adulthood.
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