• Journal of Microbiology
• /
• v.45 no.6
• /
• pp.515-520
• /
• 2007

Vaginal lactic acid-producing bacteria of 80 pre-menopausal women were studied by isolation on Blood and DeMan-Rogosa-Sharpe agar, PCR with group-specific primers for Lactobacillus-denaturing gradient gel electrophoresis (DGGE), and PCR with specific primers for V3 region in 16S rRNA-temporal temperature gel electrophoresis (TTGE). Conventional isolation method on media detected only one lactobacillus (Lactobacillus brevis) while TTGE detected only Lactobacillus sp. DGGE detected seven Lactobacillus species; L. coleohominis, L. crispatus, L. iners, L. reuteri, L. rhamnosus, L. vaginalis, and Leuconostoc lactis. L. acidophilus and L. gasseri, which are prevalent in Western women, were not detected in Korean women. Furthermore, L. rhamnosus, Leuc. lactis, L. coleohominis, and Weissella cibaria, which were not previously reported in the vaginal microbiota of Korean women, were detected. The five most prevalent LABs in vaginal microbiota in Korean women were L. iners, Enterococcus faecalis, L. crispatus, Leuc. lactis, and W. cibaria.

• Korean Journal of Poultry Science
• /
• v.27 no.1
• /
• pp.31-35
• /
• 2000

This study was investigated to observe Lactobacillus spp. population of poultry digestive organ, duodenum, ilium, cecum and colon by growing steps and to select of valuable lactobacilli as probiotics. Two strains of male broiler commercial chickens, Hybrid and Ross, were used to evaluate population of Lactobacillus spp. in intestinal tracts. Three strains of Lactobacillus were identified, and bile salts environment. The number of lactovacilli was the lowest in duodenum compared to other intestinal tracts which had similar population. Population of Lactobacillus was maintained constantly regardless growing steps after one week of age. Identification of Lactobacillus from cecum resulted in L.reuteri BC5, L. crispatus BC7, L.reuteri BC9. All strains was depressed in pH 1 and 2, although two strains could survive for one hour at pH 2. And they could survive at pH 4 for 4hours. In bile salts tolerance, L.reuteri BC5, L.crispatus BC7 were maintained for 2 hours, but the growth reduced from 2hours. Growth of L.reuteri BC9 was increased continuously. In conclusion, Lactobacillus of intestinal tracts were established at first week, and maintained constant population. They were influenced on severe acidic condition and bile salts. Cecal Lactobacillus has different growth charcteristics by strains.

• Korean Journal of Microbiology
• /
• v.48 no.4
• /
• pp.309-313
• /
• 2012

The purpose of this study is to simultaneously detect and identify specific genus of Lactobacillus distributed in vagina of healthy women in their twenties by using multiplex primer. Vaginal fluid samples were taken from 166 women who were healthy and did not have vaginosis symptoms caused by bacterial infection. Six species of lactic acid bacteria belong to Lactobacillus genus that are known to inhabit in healthy vagina were selected to make a species-specific multiplex primer. Multiplex primer I was specified to selectively detect L. iners, L. crispatus, L. gasseri and multiplex primer II was specified to selectively detect L. acidophilus, L. jensenii, L. fermentum. L. crispatus (77%) was most frequently detected, and L. acidophilus (57%) and L. jensenii (57%) were relatively higher than others. Although the proportion of L. iners (59%) was a little higher than those of L. acidophilus and L. jensenii, this should be further validated in healthy women's vagina to be sure since they often appear in women having bacterial vaginosis and/or during the antibiotic therapy. Conclusively, the multiplex PCR technique using the species-specific primer could a useful tool to predict variation of vaginal health condition and process of recovery from vaginal disease.

• Journal of Animal Science and Technology
• /
• v.50 no.5
• /
• pp.667-676
• /
• 2008

This experiment was conducted to investigate the effects of dietary supplementation of various Lactobacilli strains on growth performance, intestinal microflora, ileal villi development and ileal mucosal surface of broiler chickens. Six hundred 1-d-old male chicks(Avian) were randomly divided into five groups of four replicates, each replicates containing 30 birds and fed corn-soybean meal diets containing Latobaillus were isolated from cecum of chichens at 107cfu/g diet for five weeks. The treatments were control(antibiotics-free diet), Lactobacillus crispatus avibro1(LCB), Lactobacillus reuteri avibro2(LRB), Lactobacillus crispatus avihen1(LCH) or Lactobacillus vaginalis avihen2(LVH). The body weight gain in groups fed Lactobacillus significantly improved as compared to those of control(P<0.05). Feed intake was not statistically different among the groups. The number of Lactobacillus in ileum and cecum of chicks fed various Lactobacillus tended to be increased or significantly increased as compared to those of the control(P<0.05), but there was difference by age of chicks and species of Lactobacillus. The number of yeast was significantly increased in cecum and ileum at three weeks old chicks fed Lactobacilli compared with the control(P<0.05). The anaerobes’ number of ileum and cecum tended to increase or significantly increased in Lactobacillus treatments compared with the control(P<0.05). The ileal villi length extended greatly at three weeks of age in groups fed Lactobacillus compared with the control(P<0.05). The length of ileal villi in chicks fed Lactobacillus was continuously increased up to five weeks of age, but did not increased in the control(P<0.05). Lactobacillus was found on ileal mucosal surface. And ileal mucosal surface was maintained better with Latobacillus feeding. These results suggest the possibility that cecal Lactobacillus of chickens could be used as probiotics by improving the growth performance and promoting development of ileal villi in broiler chicks.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.918-922
• /
• 2001

In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2002.11a
• /
• pp.64-84
• /
• 2002

These studies were conducted to evaluate the Properties of lactic acid Producing bacteria(LAB), isolated from broiler and laying hens cecum and select the optimum strains to improve the performance, environment of poultry house, immunity, and intestinal microflora of broiler and laying hens. In experiment I , 23 LAB strains were isolated from broiler and laying hens cecum as a colony form. Six strains were selected by acid tolerance, bile salt tolerance, viability, enzyme release, antagonism, and antibiotics susceptibility. In Experiment II, selected LABs from Ex. 1 were conducted to investigate the effects of feeding various Lactobacillus on performance, nutrients digestibility, intestinal microflora, villi development and observation of epithelium surface, blood chemicals and fecal noxious gas of broiler chicks. One thousand eighty one day old broiler chicks were fed into Lactobacillus crispatus avibrol(LCB), Lactobacillus reuteri avibro2(LRB), Lactobacillus crispatus avihen1(LCH), and Lactobacillus vaginalis avihen2(LVH) at the level of 10$^4$ and 10$\^$7/cfu/g diet. Weight gam of chicks fed Lactobacillus tended to increase from the first week and was higher from 50 to 100g in Lactobacillus treatments than control. Feed intake and feed conversion were not statistically different of all treatments. Dry Matter digestibility of Lactobacillus treatments was prone to improve compared to that of control, but was not significantly different. Protein and Ca digestibility were also tended to improve in Lactobacillus treatments relative that of control. Lactobacillus treatments showed improved tendency in crude ash and fat compared to those of control, whereas phosphorus digestibility was not consistency. Nutrients digestibilities of bird fed LCH were superior to those of other treatments, It showed significantly higher in Ca and P digestibility than control(P〈0.05). Total Lactobacillus spp. of birds fed various Lactobacillus was significantly higher in illeum for five weeks(P〈0.05), but was not different at cecum. Yeast was thought to be not completely attached to intestinal lumen for one week. However, total number of yeast was significantly increased in cecum and illeum of three weeks old chicks (P〈0.05). The number of anaerobes exhibited to tendency the increase in Lactobacillus treatments from one week old of age at both ileum and cecum.

• Korean Journal of Poultry Science
• /
• v.28 no.1
• /
• pp.27-40
• /
• 2001

This study was conducted to investigate the effects of feeding various Lactobacillus on performance, nutrients digestibility, intestinal microflora, and fecal noxious gas of broiler chicks. One thousand eighty one dat old male broiler chicks were fed into Lactobacillus crispatus avibrd 1(LCB), Lactobacillus reuteri avibro 2(LRB), Lactobacillus crispatus avihen 1(LCH), and Lactobacillus vaginalis avehen 2(LVH) at the level of $10^{4}$ and 10(sup)7cfu/g diet. There were four replicates with thirty chicks per pen. Diets contained ME 3,100, 3,100kcal/kg, and CP 22.0, 20.0% for starting and finishing period, respectively. Weight gain of checks fed Lactobacillus spp. tended to increase from the first week and was higher from 50 to 100g in Lactobacillus treatments than control. Feed intake and feed conversion were not statistically different of all treatments. Dry Matter(DM) digestibility of Lactobacillus treatments was tended to improve compared to that of control, but was not significantly different. Protein and Ca digestibility were also tended to improve in Lactobacillus treatments relative that of control. Lactobacillus treatments showed improved tendency in crude ash and fat compared to those of control, whereas phosphorus digestibility was not consistency. Nutrients digestibilities of bird fed LCH were superior to those of other treatments. It showed significantly higher in Ca and P digestibility than control(P〈0.05). Total Lactobacillus spp. of birds fed various lactobacillus was significantly higher in ileum for five weeks(P〈0.05), but was not different at cecum. Yeast was thought to be not completely attached to intestinal lumen for one week. However, total number of yeast was significantly increased in cecum and ileum from three weeks old chicks(P〈0.05). The number of anaerobes exhibited to tendency the increase in Lantobacillus treatments from one week old of age at both ileum and cecum. Moisture contents of birds fed Lactobacillus spp. was shown from 27 to 30% at five weeks old, whereas it was 38.2% in control. It decreased approximately 25% in Lactobacillus treatments relative to that of control and showed significance between them(P〈0.05). Fecal $NH_{3}$ gas emission was significantly decreased in Lactobacillus treatments and maintained $\frac{1}{3}$to $\frac{1}{2}$ of control(P〈0.05). LCH and LVH treatments were most effective to decrease fecal $NH_{3}$ gas and especiallt impreved ti significantly at the level of the level of 10(sup)7cfu/g diets covpared to that of $10^{4}$ cfu/g diets and control(P〈0.05). In conclusion, feeding Lactobacillus spp. increase performance and nutrients digestibilities of broiler chicks and improve environments of house. Proper supplemental level of Lactobacillus spp. is considered more than $10^{4}$ cfu/g diet.

• Korean Journal of Poultry Science
• /
• v.29 no.3
• /
• pp.213-223
• /
• 2002

This study was conducted to investigate the influence of feeding various Lactobacillus on production performance, nutrients digestibility, intestinal microflora, and fecal $NH_{3}$ gas emission in laying hens. Three hundred and sixty ISA Brown layers, 21 weeks of age, were randomly allotted to nine treatments, with low replicates per treatment. Nine treatments consisted of Control(no Lactobacillus), Lactobacillus crispatus avibrol(LCB: KFCC-11195), Lactobaciilus reuteri avibro2(LRB: KFCC-11196), Lactobacillus crispatus avihenl(LCH: KFCC-11197), Lactobacillus vaginalis avihen2(LVH: KFCC-11198). Each Lactobacillus was added at two levels ($10^{4}$and $10^{7}$ cfu/g diets). Egg production, and egg weight were measured daily. A metabolism trial was conducted following the 12-week feeding trial, during which egg qualities, intestinal microflora and fecal $NH_{3}$ gas emission were examined. Egg production and daily egg mass improved significantly by the addition of various Lactobacillus(P<0.05), of which effect was more notable during the latter part of the feeding trial. But, no significant differences were found among Lactobacillus strains and between two levels of supplementation. Egg weight and feed intake showed no difference among all treatments. Feed conversion ratio of birds fed lactobacillus was significantly improved compared to that of the Control(P<0.05), but not different among lactobacillus treatments. Digestibility of crude protein, ether extract and crude ash improved significantly in lactobacillus treatments(P<0.05). However, there were not statistically different by adding levels and strains. Total counts of Lactobacillus spp. in ileum of layers fed Lactobacillus were significantly higher than that of the control, but no consistent trend was found in cecum. There were no significant differences in intestinal yeast and anaerobes counts among all treatments. The Lactobaciilus supplementation did not exert my effect on the eggshell quality and Haugh unit. Fecal $NH_{3}$ gas emission decreased significantly in Lactobacillus treatments, and showed no difference between the two supplementation levels. From the result of this study, it could be concluded that dietary supplementation of Lactobacillus, regardless of their species, Improves the laying performance and decreases the fecal ammonia gas emission. The proper level of supplementation appears to be $10^{4}$ cfu/g of diet.

• KSBB Journal
• /
• v.17 no.3
• /
• pp.311-313
• /
• 2002

Lactobacillus spry. are Gram-positive bacteria playing important roles in human health. In this study, we successfully isolated the total RNA from the cells broken by glass beads using hot phenol method. Moreover, we were able to omit lysozyme and proteinase K treatment by using glass beads to break cell more efficiently. This method was more rapid and simple when compared to the previous one. Prepared RNA can be used for the transcriptional analysis of Lactobacillus spp.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.2
• /
• pp.248-258
• /
• 2020

The vaginal microbiota may be important for pregnancy prognosis because vaginal dysbiosis during pregnancy appears to be related to preterm birth (PTB) or pregnancy loss. Previous reports have indicated that a Lactobacillus-poor microbial flora in the vagina and intrauterine infection by diverse anaerobes ascending from the vagina are associated with undesirable delivery outcomes. However, no research has involved the use of pyrosequencing analysis to examine vaginal microbiota profiles or their potential associations with high-risk pregnancy in Korean women. Vaginal swabs were collected from 500 Korean women for the identification of community state types (CSTs). Of these, 137 samples were further analyzed using a Roche/454 GS Junior pyrosequencer. Three distinct CSTs were identified based on the dominant vaginal microbes: CST I (Lactobacillus crispatus dominated), CST III (Lactobacillus iners dominated), and CST IV (with diverse species of anaerobes). Twelve of the 67 pregnant women had undesirable pregnancy outcomes (four miscarriages and eight PTBs). The dominant microbe in the vaginal microbiota of women who gave birth at full-term was L. crispatus. In contrast, L. iners was the dominant vaginal microbe in women who miscarried. Most (n = 6/8) vaginal microbiota profiles of women who experienced PTB could be classified as CST IV, with diverse bacteria, including anaerobic vaginal species. The present study provides valuable information regarding the characteristics of the vaginal microbiota of Korean women related to high-risk pregnancy. Investigation of the vaginal microbiotic structure in pregnant Korean women is necessary to enable better prediction of adverse pregnancy outcomes.