Regulation of serotonin production by specific microbes from piglet gut
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Serotonin is an important signaling molecule that regulates secretory and sensory functions in the gut. Gut microbiota has been demonstrated to affect serotonin synthesis in rodent models. However, how gut microbes regulate intestinal serotonin production in piglets remains vague. To investigate the relationship between microbiota and serotonin specifically in the colon, microbial composition and serotonin concentration were analyzed in ileum-cannulated piglets subjected to antibiotic infusion from the ileum when comparing with saline infusion. Microbes that correlated positively with serotonin production were isolated from piglet colon and were further used to investigate the regulation mechanisms on serotonin production in IPEC-J2 and a putative enterochromaffin cell line RIN-14B cells.Antibiotic infusion increased quantities of Lactobacillus amylovorus (LA) that positively correlated with increased serotonin concentrations in the colon, while no effects observed for Limosilactobacillus reuteri (LR). To understand how microbes regulate serotonin, representative strains of LA, LR, and Streptococcus alactolyticus (SA, enriched in feces from prior observation) were selected for cell culture studies. Compared to the control group, LA, LR and SA supernatants significantly up-regulated tryptophan hydroxylase 1 (TPH1) expression and promoted serotonin production in IPEC-J2 cells, while in RIN-14B cells only LA exerted similar action. To investigate potential mechanisms mediated by microbe-derived molecules, microbial metabolites including lactate, acetate, glutamine, and γ-aminobutyric acid were selected for cell treatment based on computational and metabolite profiling in bacterial supernatant. Among these metabolites, acetate upregulated the expression of free fatty acid receptor 3 and TPH1 while downregulated indoleamine 2,3-dioxygenase 1. Similar effects were also recapitulated when treating the cells with AR420626, an agonist targeting free fatty acid receptor 3.Overall, these results suggest that Lactobacillus amylovorus showed a positive correlation with serotonin production in the pig gut and exhibited a remarkable ability to regulate serotonin production in cell cultures. These findings provide evidence that microbial metabolites mediate the dialogue between microbes and host, which reveals a potential approach using microbial manipulation to regulate intestinal serotonin biosynthesis.Keywords:
Enterochromaffin cell
Lactobacillus reuteri
Objective To study the composition of Lactic acid bacteria in the intestine of healthy ICR mice.Methods Fresh fecal samples from 10 healthy male ICR mice were collected and the total genomic DNA was extracted from each sample.Lactic acid bacteria(LAB) group specific primer(Lac1) and universal primer for bacteria(1391r) were used to amplify 16S rRNA gene and construct LAB group specific clone library to analyze the species composition of LAB.Results The results of clone library showed that Lactobacillus reuteri and Lactobacillus johnsonii were the most predominant species in gut of mice.Lactobacillus murinus,Lactobacillus vaginalis,Lactobacillus intestinalis and a potentially new species were also detected.Conclusion The composition and structure of Lactobacillus in healthy ICR mice have high diversity and the species of Lactobacillus reuteri may have high microdiversity at strain level.
Lactobacillus reuteri
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clone (Java method)
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Effect of hot water extract (HWE) from Fructus tribuli on growth of Lactobacillus casei , Lactobacillus reuteri , Lactobacillus rhamnosus and Lactobacillus bulgaricus with material properties was studied by measuring optical density at 600nm (OD 600 ) and pH using MRS media as the control. The addition of HWE (v/v) was 1%, 2%, 3%, 4% and 5%. Results were as follows: addition of HWE has the significant inhibition on growth of Lactobacillus reuteri and Lactobacillus bulgaricus and the significant promotion on growth of Lactobacillus casei and Lactobacillus rhamnosus . The optimum concentration of HWE in MRS media was 2% for Lactobacillus casei and 3% for Lactobacillus rhamnosus , respectively.
Lactobacillus casei
Lactobacillus reuteri
Lactobacillus rhamnosus
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Modern molecular genetic methods of research, which emerged in the 21st century, completely unfixed all established notions not only about the microcosm, but also about its influence on the life and health of the macroorganism. The identification of intestinal bacteria and particularities of their various strains is increasingly expanding our understanding of the mechanisms of their action on the somatic and mental health of a human being. The various lactobacilli strains have been the most studied probiotic at the present time. The genus Lactobacillus is known to include about 90 species with a wide range of biochemical and physiological properties. Lactobacillus reuteri, which has strains with different mechanisms of action, is one of the unique microorganisms of the family Lactobacillaceae. The range of possibilities for using various Lactobacillus reuteri strains is currently very wide. This article deals with clinical effects associated with the strain-specificity of Lactobacillus reuteri. Современные молекулярно-генетические методы исследования, появившиеся в XXI в., полностью перевернули наши представления не только о микромире, но и о его влиянии на жизнь и здоровье макроорганизма. Идентификация кишечных бактерий и особенности их разных штаммов все больше расширяют наше понимание механизмов их влияния на соматическое и психическое здоровье человека. На сегодняшний день наиболее изученными являются различные штаммы лактобацилл. Известно, что род Lactobacillus включает примерно 90 видов с широким спектром биохимических и физиологических свойств. Одним из уникальных микроорганизмов семейства Lactobacillaceae является Lactobacillus reuteri, имеющая штаммы с разным механизмом действия. На сегодняшний день спектр возможностей использования разных штаммов Lactobacillus reuteri очень широкий. В данной статье рассмотрены клинические эффекты, связанные со штаммоспецифичностью Lactobacillus reuteri.
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To analyze the microbiota shift in the distal esophagus of Sprague-Dawley rats fed a high-fat diet.Twenty Sprague-Dawley rats were divided into high-fat diet and normal control groups of 10 rats each. The composition of microbiota in the mucosa from the distal esophagus was analyzed based on selective culture. A variety of Lactobacillus species were identified by molecular biological techniques. Bacterial DNA from Lactobacillus colonies was extracted, and 16S rDNA was amplified by PCR using bacterial universal primers. The amplified 16S rDNA products were separated by denaturing gradient gel electrophoresis (DGGE). Every single band was purified from the gel and sent to be sequenced.Based on mucosal bacterial culturing in the distal esophagus, Staphylococcus aureus was absent, and total anaerobes and Lactobacillus species were decreased significantly in the high-fat diet group compared with the normal control group (P < 0.01). Detailed DGGE analysis on the composition of Lactobacillus species in the distal esophagus revealed that Lactobacillus crispatus, Lactobacillus gasseri (L. gasseri) and Lactobacillus reuteri (L. reuteri) comprised the Lactobacillus species in the high-fat diet group, while the composition of Lactobacillus species in the normal control group consisted of L. gasseri, Lactobacillus jensenii and L. reuteri.High-fat diet led to a mucosal microflora shift in the distal esophagus in rats, especially the composition of Lactobacillus species.
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Lactobacillus crispatus
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The acid ureases from 37 strains belonging to the genera Lactobacillus and Streptococcus were partially purified and characterized. The 37 acid ureases were classified into 6 different types according to their electrophoretic mobilities. The 6 different types of acid ureases, from 6 representative strains (4 species of Lactobacillus and 2 species of Streptococcus), were further characterized. The molecular sizes of the enzymes ranged from 140, 000 for Streptococcus salivarius to 350, 000 for Lactobacillus animalis. The isoelectric points were found to be 4.6 (Streptococcus mitior), 4.7 (Lactobacillus reuteri, Lactobacillus ruminis and Streptococcus salivarius) and 4.8 (Lactobacillus animalis and Lactobacillus fermentum). The optimum pHs ranged from 2.0 for Lactobacillus fermentum and Lactobacillus reuteri to 5.0 for Lactobacillus ruminis. The optimum temperatures ranged from 55°C for Lactobacillus animalis and Lactobacillus ruminis to 65°C for Lactobacillus fermentum, Lactobacillus reuteri and Streptococcus mitior. All 6 different acid ureases eliminated urea in alcoholic beverages that are generally acidic.
Lactobacillus fermentum
Lactobacillus reuteri
Lactobacillus salivarius
Streptococcus salivarius
Lactobacillus brevis
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Ageing is associated with important changes in gastrointestinal function and in the levels of intestinal hormones secreted. Enterochromaffin (EC) cells containing serotonin (5-HT) and melatonin may play a major role in maintaining gut function during ageing. Our aim was to characterise the mucosal availability of 5-HT and melatonin in the ileum and colon of a mouse model of ageing. Female young mice (2-5 month; n = 6), aged mice (22-24 months; n = 6) and aged mice treated with melatonin (n = 6; 10 mg/kg/day) were examined. Electrochemical methods were used to measure 5-HT and melatonin concentrations near the mucosal surface of ileum and distal colon. Amperometry studies showed that steady state levels of 5-HT from ileum and colon were decreased in aged mice treated with melatonin when compared to aged mice, while compression-evoked 5-HT release was unchanged. Differential pulse voltammetry studies showed that young mice had concentrations of 5-HT of 4.8 +/- 0.8 mum in the ileum and 4.9 +/- 1.0 mum in the colon. Concentrations of melatonin were 5.7 +/- 1.4 mum in the ileum and 5.6 +/- 1.9 mum in the colon. Compared to young mice, the levels of 5-HT and melatonin were increased in aged mice (combined ileum and colon: 5-HT = 130% and melatonin = 126% of young mice) and decreased in melatonin-treated mice (5-HT = 94% and melatonin = 82%). In conclusion, our data show that the availability of gut 5-HT and melatonin is increased in aged mice and melatonin treatment suppresses natural gastrointestinal production of 5-HT and melatonin in the aged mouse intestine.
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Lactobacillus reuteri
Prebiotic
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To develop species-specific monitoring techniques for rapid detection and identification of Lactobacillus isolated from mouse faeces.The specificity of oligonucleotide probes was evaluated by dot blot hybridization to 16S rDNA and 23S rDNA amplified by PCR from 12 Lactobacillus type strains and 100 strains of Lactobacillus isolated from mouse faeces. Oligonucleotide probes specific for each Lactobacillus species hybridized only with targeted rDNA. The Lactobacillus strains isolated from mouse faeces were identified mainly as Lactobacillus intestinalis, L. johnsonii, L. murinus and L. reuteri using species-specific probes. 16S rDNA of eight unidentified isolates were sequenced and two new probes were designed. Four of eight strains of unhybridized Lactobacillus were identified as L. johnsonii/gasseri group, and the remaining four strains as L. vaginalis.The species-specific probe set of L. intestinalis, L. johnsonii, L. murinus, L. reuteri and L. vaginalis in this study was efficient for rapid identification of Lactobacillus isolated from mouse faeces.The oligonucleotide probe set for Lactobacillus species harboured in the mouse intestine, can be used for rapid identification of lactobacilli and monitoring of the faecal Lactobacillus community.
Lactobacillus reuteri
Lactobacillus gasseri
Lactobacillus crispatus
Oligomer restriction
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سویههای پروبیوتیکی Lactobacillus قابلیت استفاده به عنوان نگهدارنده غذایی را دارند. پژوهش حاضر با هدف بررسی استفاده از Lactobacillus delbrueckii subsp. bulgaricus (PTCC 1737)، L. reuteri (PTCC 1655)، L. brevis CD0817 و ترکیب باکتریهای Lactobacillus روی کیفیت میکروبی فیله قزلآلای رنگینکمان در یخچال و انجماد انجام شد. تیمارها شامل فیلههای غوطهور شده در سوسپانسیون L. reuteri، L. brevis، L. delbrueckii و ترکیب آنها بودند. فیلههای غوطهور شده در اسید لاکتیک ۲ درصد و فیلههای بدون افزودنی به عنوان تیمارهای شاهد در نظر گرفته شدند. تیمارها به مدت ۵ روز در یخچال و شش ماه در انجماد نگهداری شدند. Staphylococcus aureus و Escherichia coli در تیمارهای آزمایشی و شاهد مشاهده نشدند.L. delbrueckii قادر به بقا تحت شرایط اتجماد نبود. تعداد Lactobacillusها در تیمارهای ترکیبی طی مدت زمان نگهداری در شرایط انجماد کاهش داشت (logCFU/g۳۴/۱). در L. brevis اسیدهای چرب اشباع ۹۱/۲۳ درصد و غیراشباع ۷۷ درصد و در تیمار L. reuteri اسیدهای چرب اشباع ۱۲/۲۷ درصد و غیراشباع ۸۸/۷۲ درصد بود (05/0P<). افزایش معنیدار در نسبت اسیدهای چرب غیراشباع به اشباع در L. brevis در مقایسه باL. reuteri و عدم تفاوت معنیدار در تعداد L. brevis طی مدت زمان نگهداری در دمای انجماد سبب شد که L. brevis به عنوان محافظ غذایی برای نگهداری فیله قزلآلای رنگینکمان پرورشی در یخچال و انجماد پیشنهاد شود.
Lactobacillus reuteri
Lactobacillus brevis
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