Lactobacillus acidophilus UBLA-34, L. paracasei UBLPC-35, L. plantarum UBLP-40, and L. reuteri UBLRU-87 were isolated from different varieties of fermented foods. To determine the probiotic safety at the strain level, the whole genome of the respective strains was sequenced, assembled, and characterized. Both the core-genome and pan-genome phylogeny showed that L. reuteri was closest to L. plantarum than to L. acidophilus, which was closest to L. paracasei. The genomic analysis of all the strains confirmed the absence of genes encoding putative virulence factors, antibiotic resistance, and the plasmids.
PARK JAE-YONG;NAM SU JIN;KIM JONG-HWAN;JEONG SEON-JU;KIM JUNG KON;HA YEONG LAE;KIM JEONG HWAN
Journal of Microbiology and Biotechnology
/
v.15
no.3
/
pp.525-531
/
2005
From a genomic library of Lactobacillus reuteri ATCC 55739, one clone, NE347, carrying a pyrH gene encoding UMP kinase, was identified. pNE347 carried a 1.88 kb EcoRI fragment and the pyrH was located in the middle of the insert. pyrH ORF was 723 bp in size and capable of encoding UMP kinase composed of 240 amino acid residues. tsf encoding an elongation factor-Ts and frr encoding a ribosomal recycling factor were present upstream and downstream of pyrH, respectively. When introduced into E. coli KUR1244, a pyrH-negative strain, pNE347 restored the ability to grow at $42^{\circ}C$, indicating that pyrH from L. reuteri synthesized functional UMP kinase in E. coli. Northern blot experiment showed that pyrH and frr were cotranscribed as a 1.4 kb single transcript. pyrH was overexpressed in E. coli by using a pET26b(+) vector, and a major 25 kDa protein band appeared on SDS-polyacrylamide gel.
This study was conducted to evaluate the feasibility of using L. reuteri Pg4, a strain isolated from the gastrointestinal (GI) tract of healthy broilers, as a probiotic. In preliminary in vitro studies the Pg4 strain was proven capable of tolerating acid and bile salts, inhibiting pathogenic bacteria and can adhere to intestinal epithelial cells. The probiotic properties were then evaluated on the basis of the broiler's growth performance, intestinal microbial population and cecal volatile fatty acid and lactic acid concentrations under conventional feeding. Dietary supplementation of dried L. reuteri Pg4 decreased significantly feed intake in grower chickens and improved significantly the feed conversion by 5% in a 0-6 weeks feeding period compared with the control group. The Lactobacillus counts in the crop, ileum, and cecum of the probiotic group were higher than in the control group. The L. reuteri Pg4 strain was traceable in the GI tract of probiotic supplemented chicks and showed capability of survival in the intestine for a protracted period. The probiotic group had a higher lactic acid concentration and lower pH value in the cecum than the control chicks. Probiotic supplement also affected the histology of the intestinal mucosa of chicks. The present findings demonstrated that L. reuteri Pg4 possesses probiotic characteristics and it is suggested, therefore, that the organism could be a candidate for a new probiotic strain.
A feeding trial was conducted to investigate the influence of feeding Lactobacillus reuteri culture (LR) on productive performance, intestinal microflora and availability in laying hens. Four hundred and eighty, Isa-Brown layers, 49 weeks of age, were fed diets supplemented with LR at the level of 0 (control), 0.1, 0.2, and $0.4\%$ of the diets for eight weeks. Egg production and egg weight were measured daily. Feed intake was weighed every two weeks. Egg quality was measured three times at the start, mid-term, and end of the experiment. Intestinal microflora were examined for Lactobacillus spp., E. coli and Salmonella at the end of the experiment. Overall egg production was the highest in $0.2\%$ LR (P<0.05), but that of $0.1\%$ or $0.4\%$ LR treatments did not significantly differ from that of control. Egg weight was significantly higher in LR feeding group than the control (P<0.05). Daily egg mass was significantly higher in $0.2\%$ and $0.4\%$ LR treatments compared to the control and $0.1\%$ LR (P<0.05). The number of jumbo and extra large eggs were increased in LR supplemented groups, especially in $0.1\%$ LR. Feed intake of layers fed LR supplemented diets tended to be lower than the control. However, feed conversion ratio significantly improved in LR supplemented groups (P<0.05). Availability of dry matter and crude protein improved significantly in $0.4\%$ LR treatment (P<0.05). But, those of ether extract and crude ash were not significantly different among treatments. Eggshell breaking strength and eggshell thickness were not significantly influenced by LR supplementation, and Haugh unit and yolk index were also similar to the control. Total number of Lactobacillus spp. in ileum and cecum fed LR supplemented diets were significantly higher than those of the control (P<0.05). There were no significant differences in intestinal E. coli and Salmonella in all treatments. Therefore, it is concluded that dietary supplementation of Lactobacillus reuteri culture can improve the laying performance, feed efficiency and intestinal Lactobacillus.
A feeding trial was carried out to evaluate the effects of feeding Lactobacillus reuteri culture(LRC) on the performance, nutrients digestibility, intestinal microflora, serum metabolites, ammonia generation and litter dampness in broiler chicks. Four hundred eighty, one day old male chicks were fed into none, 0.1, 0.2 and 0.4% of LRC supplementation fer seven weeks. Basal diets excluded antibiotics contained ME 3,100, 3,100 kcal/kg, and CP 22.0, 20.0% for starter and grower, respectively. Weight gain of chicks fed LRC was significantly higher than no supplemental group in overall period(P<0.05). Feed intake was the highest in the 0.1% LRC, but not statistically different from other treatments. Feed conversion showed no significance among treatments. Viable Lactobacillus spp. number of chicks fed 0.2 and 0.4% LRC was significantly higher in cecum at seven weeks of age compared to the none(P<0.05). The tendency of anaerobes number was similar to Lactobacillus spp in ileum and cecum. Total number of E. coli and Salmonella were no difference in all treatments. In serum metabolites, feeding LRC increased triglyceride, and inorganic phosphorus, but no different total protein, albumin, total cholesterol, glucose, blood urea nitrogen and Ca. Nutrients digestibility improved significantly in 0.4% LRC compared to that of none(P<0.05). Fecal NH$_3$, gas generation was greatly decreased in the LRC supplemental groups(P<0.05). Moisture contents of bedding was also significantly decreased in LRC feeding group. It was concluded from the present study that feeding Lactobacillus reuteri culture improved the growth performance and nutrients digestibility of broiler chicks and minimize the fecal noxious gas emission.
Proceedings of the Korea Society of Poultry Science Conference
/
2003.11a
/
pp.105-106
/
2003
This study was conducted to investigate the effects of supplemental Lactobacillus with drinking water on growing performance, nutrients digestibility, intestinal microflora, fecal NH3-N and VFA of broiler chicks. Four hundreds and eighty one day broiler chicks were divided into no supplement (none) and three drinking water, 10$^{6}$ (T1), 10$^4$(T2) and 10$^2$ cfu/ml water(T3) with Lactobacillus reuteri for five week. Weight gain of T2 was the highest in all treatments(P<0.05), and other supplements also tended to increase but there were not significantly difference compared to the none. Feed intake and feed conversion were not statistically different of all treatments. Dry Matter digestibility of Lactobacillus treatments was tended to improve compared to that of none, but was not significantly different. Protein and fat digestibility were also tended to Improve In Lactobacillus treatments relative that of control(P<0.05), whereas ash digestibility was not consistency. Total Lactobacillus spp. of birds fed Lactobacillus was significantly higher in cecum at five weeks(P<0.05), but was not different at ileum. E. coli and Salmonella were not difference by treatments. In conclusion, feeding Lactobacillus increase performance and nutrients digestibility of broiler chicks and Improve environments of house.
Journal of the Korea Academia-Industrial cooperation Society
/
v.20
no.12
/
pp.110-116
/
2019
To investigate the antagonistic inhibitory effects in a mixed culture between probiotics and various pathogenic microorganisms, 140 probiotics were identified using a 16 rRNA sequencing phylogenetic analysis method, and various probiotics strains were isolated from Korean kimchi from January to December 2016. The antagonistic inhibition test of a mixed culture of four probiotics (Enterococcus faecalis, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus reuteri) with excellent antimicrobial activity and six pathogenic microorganisms (Candida albicans, Salmonella Enteritidis, E. coli O157:H7, Shigella flexneri, Staphylococcus aureus, and Pseudomonas aeruginosa)showed that the growth of most probiotics strains increased normally after culture, but growth was inhibited almost completely in most pathogenic microorganisms, except for S. Enteritidis. This antagonistic inhibitory effect in vitro was attributed to the low pH of the lactic acid and organic acid produced during fermentation. As a result, four probiotics strains isolated from Korean Kimchi are very likely to be developed as therapeutic agents for female yeast infections and colon and skin care. In the future, these therapeutic agents will help improve public health related to probiotics.
Kim, Dong-Wook;Kim, Ji-Hyuk;Kang, Geun-Ho;Kang, Hwan-Ku;Lee, Sang-Jin;Lee, Won-Jun;Kim, Sang-Ho
Journal of Animal Science and Technology
/
v.50
no.6
/
pp.807-818
/
2008
This study was conducted to prove the optimum feeding method of Lactobacillus in broiler chickens by investigating the intestinal viability of ingested Lactobacillus and the effect of feeding levels and frequency of Lactobacillus on growth performance in broiler chickens. In experiment 1, A total of one hundred, 5 weeks old male broiler chickens(Abor Acre) were fed Lactobacillus reuteri avibro2 expressed green fluorescent protein(GFP) at 104cfu/g diet to investigate the retention time of ingested Lactobacillus in the intestine for 1 day. The percentage of Lactobacillus expressed GFP in intestinal contents was 26% at 1 day after fed Lactobacillus expressed GFP. The percentage of Lactobacillus expressed GFP in intestinal contents was decreased in length of time. In experiment 2, A total of four hundred eighty, 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each to prove the optimum feeding level of Lactobacillus. The treatments were control(free antibiotics), Lactobacillus reuteri avibro2 5.0×10cfu/mL, 5.0×103cfu/mL, and 5.0×105cfu/mL. The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). Feed conversion ratio was not significantly difference among the groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). Protein and fat digestibility in Lactobacillus 5.0×103cfu/mL and 5.0×105cfu/mL treated groups were significantly improved(P<0.05). No significant differences were observed on the availability of dry matter and crude ash in Lactobacillus treatments compared to those of control. In experiment 3, A total of six hundred 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each and were fed Lactobacillus reuteri avibro2 at intervals of 1, 2, 3, and 5 day for five weeks. Feeding level of Lactobacillus was 5.0×103cfu/mL The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). The final body weight and body weight gain were significantly increased, when Lactobacillus was fed at intervals of 1 days, or 2 days. There were no significant differences in feed intake and feed conversion ratio among the all groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). No significant differences were observed on the number of coliform bacteria and Salmonella of ileum and cecum. Consequently, supplemental Lactobacillus influenced positive effects on the growth performance, nutrient availability and intestinal microflora. The optimum feeding level of Lactobacillus was 5.0×103cfu/mL, and the constant feeding of Lactobacillus was effective.
Purpose: Gastrointestinal symptoms are often related to antibiotic treatment. Their incidence, risk and protective conditions in children are not well defined and represent the aims of this study. Methods: We prospectively enrolled inpatient children submitted to antibiotic treatment. Indication, type, dose and duration of treatment, probiotic supplementation and gastrointestinal symptoms were recorded at recruitment, after two and four weeks. Antibiotic-associated diarrhea (AAD) was defined as the presence of at least 3 loose/liquid stools within 14 days from antibiotic onset. Results: AAD occurred in 59/289 (20.4%) of patients, with increased risk in children younger than 3 years (relative risk [RR]=4.25), in lower respiratory (RR=2.11) and urinary infections (RR=3.67), intravenous administration (RR=1.81) and previous AAD episodes (RR=1.87). Abdominal pain occurred in 27/289 (9.3%), particularly in children >6 years (RR=4.15), with previous abdominal pain (RR=7.2) or constipation (RR=4.06). Constipation was recorded in 23/289 (8.0%), with increased risk in children having surgery (RR=2.56) or previous constipation (RR=7.38). Probiotic supplementation significantly reduced AAD (RR=0.30) and abdominal pain (RR=0.36). Lactobacillus rhamnosus GG (LGG) and L. reuteri significantly reduced AAD (RR=0.37 and 0.35) and abdominal pain (RR=0.37 and 0.24). Conclusion: AAD occurred in 20.4% of children, with increased risk at younger age, lower respiratory and urinary tract infections, intravenous treatment and previous AAD. LGG and L. reuteri reduced both AAD and associated abdominal pain.
Probiotics are considered as the best effective alternatives to antibiotics. The aim of this study was to characterize the probiotic potential of lactobacilli for use in swine farming by using in vitro evaluation methods. A total of 106 lactic acid bacterial isolates, originating from porcine feces, were first screened for the capacity to survive stresses considered important for putative probiotic strains. Sixteen isolates showed notable acid and bile resistance, antibacterial activity, and adherence to intestinal porcine epithelial cells (IPEC-1). One isolate, LR1, identified as Lactobacillus reuteri, was selected for extensive study of its probiotic and functional properties in IPEC-1 cell models. L. reuteri LR1 exhibited good adhesion to IPEC-1 cells and could inhibit the adhesion of enterotoxigenic Escherichia coli (ETEC) to IPEC-1 cells. L. reuteri LR1 could also modulate transcript and protein expression of cytokines involved in inflammation in IPEC-1 cells; the Lactobacillus strain inhibited the ETEC-induced expression of proinflammatory transcripts (IL-6 and TNF-α) and protein (IL-6), and increased the level of anti-inflammatory cytokine (IL-10). Measurement of the permeation of FD-4 showed that L. reuteri LR1 could maintain barrier integrity in monolayer IPEC-1 cells exposed to ETEC. Immunolocalization experiments showed L. reuteri LR1 could also prevent ETEC-induced tight junction ZO-1 disruption. Together, these results indicate that L. reuteri LR1 exhibits desirable probiotic properties and could be a potential probiotic for use in swine production.
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