Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans / Disbiose intestinal, inflamação e diabetes tipo 2 em camundongos usando microbiota intestinal sintética de humanos diabéticos

Abstract The study was aimed to assess impact of high fat diet (HFD) and synthetic human gut microbiota (GM) combined with HFD and chow diet (CD) in inducing type-2 diabetes (T2D) using mice model. To our knowledge, this is the first study using selected human GM transplantation via culture based method coupled dietary modulation in mice for in vivo establishment of inflammation leading to T2D and gut dysbiosis. Twenty bacteria (T2D1-T2D20) from stool samples of confirmed T2D subjects were found to be morphologically different and subjected to purification on different media both aerobically and anerobically, which revealed seven bacteria more common among 20 isolates on the basis of biochemical characterization. On the basis of 16S rRNA gene sequencing, these seven isolates were identified as Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenes (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). The seven isolates were subsequently used as synthetic gut microbiome (GM) for their role in inducing T2D in mice. Inbred strains of albino mice were divided into four groups and were fed with CD, HFD, GM+HFD and GM+CD. Mice receiving HFD and GM+modified diet (CD/HFD) showed highly significant (P<0.05) increase in weight and blood glucose concentration as well as elevated level of inflammatory cytokines (TNF-α, IL-6, and MCP-1) compared to mice receiving CD only. The 16S rRNA gene sequencing of 11 fecal bacteria obtained from three randomly selected animals from each group revealed gut dysbiosis in animals receiving GM. Bacterial strains including Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) and Lactobacillus gasseri (MT152635) were isolated from mice treated with GM+modified diet (HFD/CD) compared to strains Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629) which were isolated from mice receiving CD/HFD. In conclusion, these findings suggest that constitution of GM and diet plays significant role in inflammation leading to onset or/and possibly progression of T2D. .

Resumo O estudo teve como objetivo avaliar o impacto da dieta rica em gordura (HFD) e da microbiota intestinal humana sintética (GM) combinada com HFD e dieta alimentar (CD) na indução de diabetes tipo 2 (T2D) usando modelo de camundongos. Para nosso conhecimento, este é o primeiro estudo usando transplante de GM humano selecionado através do método baseado em cultura acoplada à modulação dietética em camundongos para o estabelecimento in vivo de inflamação que leva a T2D e disbiose intestinal. Vinte bactérias (T2D1-T2D20) de amostras de fezes de indivíduos T2D confirmados verificaram ser morfologicamente diferentes e foram submetidas à purificação em meios diferentes aerobicamente e anaerobicamente, o que revelou sete bactérias mais comuns entre 20 isolados com base na caracterização bioquímica. Com base no sequenciamento do gene 16S rRNA, esses sete isolados foram identificados como Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenides (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). Esses sete isolados foram, posteriormente, usados ​​como microbioma intestinal sintético (GM) por seu papel na indução de T2D em camundongos. Linhagens consanguíneas de camundongos albinos foram divididas em quatro grupos e foram alimentadas com CD, HFD, GM + HFD e GM + CD. Camundongos que receberam a dieta modificada com HFD e GM + (CD / HFD) mostraram um aumento altamente significativo (P < 0,05) no peso e na concentração de glicose no sangue, bem como um nível elevado de citocinas inflamatórias (TNF-α, IL-6 e MCP-1) em comparação com os ratos que receberam apenas CD. O sequenciamento do gene 16S rRNA de 11 bactérias fecais obtidas de três animais selecionados aleatoriamente de cada grupo revelou disbiose intestinal em animais que receberam GM. Cepas bacterianas, incluindo Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) e Lactobacillus Gasseri (MT152635D), foram tratadas com dieta modificada / CD) em comparação com as linhagens Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629), que foram isoladas de camundongos recebendo CD / HFD. Em conclusão, esses resultados sugerem que a constituição de GM e dieta desempenham papel significativo na inflamação levando ao início ou/e possivelmente à progressão de T2D.

Metagenomics approach to identify lignocellulose-degrading enzymes in the gut microbiota of the Chinese bamboo rat cecum

BACKGROUND: Lignocellulose is considered a renewable organic material, but the industrial production of biofuel from lignocellulose is challenging because of the lack of highly active hydrolytic enzymes. The guts of herbivores contain many symbiotic microorganisms that have evolved to hydrolyze plant lignocellulose. Chinese bamboo rats mainly consume high-fiber foods, indicating that some members of the intestinal tract microbiota digest lignocellulose, providing these rats with the energy required for growth. RESULTS: Here, we used metagenomics to analyze the diversity and functions of the gut microbiota in Chinese bamboo rats. We identified abundant populations of lignocellulose-degrading bacteria, whose main functions involved carbohydrate, amino acid, and nucleic acid metabolism. We also found 587 carbohydrate-active enzyme genes belonging to different families, including 7 carbohydrate esterase families and 21 glycoside hydrolase families. The glycoside hydrolase 3, glycoside hydrolase 1, glycoside hydrolase 43, carbohydrate esterase 4, carbohydrate esterase 1, and carbohydrate esterase 3 families demonstrated outstanding performance. CONCLUSIONS: The microbes and enzymes identified in our study expand the existing arsenal of proficient degraders and enzymes for lignocellulosic biofuel production. This study also describes a powerful approach for targeting gut microbes and enzymes in numerous industries.

Minocycline and Its Impact on Microbial Dysbiosis in the Skin and Gastrointestinal Tract of Acne Patients

Gut microbiota in Mexican patients with common variable immunodeficiency

Introduction: Common variable immunodeficiency (CVID) is the main symptomatic primary immunodeficiency and is associated with complex immune disorders. Gut microbiota interacts closely with the immune system, and intestinal dysbiosis is related to multiple diseases. Objective: To describe for the first time the composition of gut microbiota in Mexican patients with CVID. Methods: Fecal samples from five patients with CVID were collected and massive sequencing of the V3-V4 region of 16S rRNA gene was carried out using illumina technology. Results: Bacterial relative abundance was observed at all taxonomic levels. Firmicutes, Actinobacteria and Verrucomicrobia were the predominant phyla. The Clostridia class and the Clostridial order were the most common in their respective taxon; the Ruminococcaceae family predominated. A total of 166 genera were reported, with the most abundant being Faecalibacterium. Five species were identified, but only Bifidobacterium longum was present in all patients. Conclusions: Unlike healthy subjects' gut microbiota, where Firmicutes and Bacteroidetes predominate, the microbiota of the patients with CVID considered in this study was abundant in Firmicutes, Actinobacteria and Verrucomicrobia. The low presence of Bacteroidetes and high abundance of Firmicutes might indicate the existence of intestinal dysbiosis in these patients.

Dominant bacterial and archaeal phyla associated with top soils sourced from commercial farm holding in Delta State, Nigeria

ABSTRACT: The dominant bacteriological and archaeal phyla of compounded soils sourced from a commercial farm estate located in Amukpe town and a nearby control in Adavware community both in Delta State, were evaluated with the aid of Next Generation Sequencing (NGS) protocols. The residual herbicide and pesticide composition of the bulked soils were also determined using gas chromatography (GC) and electron capture detector (ECD). Total concentrations of the extracted DNA were 6.83 and 6.65 ng/µl for the control and experimental soils. Nine (9) bacterial phyla; Proteobacteria, Actinobacteria, Chloroflexi, Firmicutes, Verrucomicrobia, Planctomycetes, Bacteroidetes Acidobacteria, and Elusimicrobia were observed in the control soil. Thirteen (13) bacterial phyla; Elusimicrobia, Fibrobacteres Lentisphaerae, Armatimonadetes, Cyanobacteria/Chloroplast, Bacteroidetes, Actinobacteria, Proteobacteria, Chloroflexi, Firmicutes, Acidobacteria, Planctomycetes and Verrucomicrobia were detected in the experimental soil. Two (2) archaeal phyla; Euryarchaeota, and Diapherotrites were detected both the experimental and control soil, whilst an additional archaeal phylum; Woesearchaeota was present in only the experimental soil. The total organochloride phosphate component of the experimental soil was 1.4µg/Kg and 0.4µg/Kg for the control soil respectively

Is stool frequency associated with the richness and community composition of gut microbiota?

BACKGROUND/AIMS: Recently, a number of studies have reported that the gut microbiota could contribute to human conditions, including obesity, inflammation, cancer development, and behavior. We hypothesized that the composition and distribution of gut microbiota are different according to stool frequency, and attempted to identify the association between gut microbiota and stool frequency. METHODS: We collected fecal samples from healthy individuals divided into 3 groups according to stool frequency: group 1, a small number of defecation (≤2 times/wk); group 2, normal defecation (1 time/day or 1 time/2 day); and group 3, a large number of defecation (≥2–3 times/day). We evaluated the composition and distribution of the gut microbiota in each group via 16S rRNA-based taxonomic profiling of the fecal samples. RESULTS: Fecal samples were collected from a total of 60 individuals (31 men and 29 women, aged 34.1±5.88 years), and each group comprised 20 individuals. The microbial richness of group 1 was significantly higher than that of group 3 and tended to decrease with increasing number of defecation (P<0.05). The biological community composition was fairly different according to the number of defecation, and Bacteroidetes to Firmicutes ratio was higher in group 1 than in the other groups. Moreover, we found specific strains at the family and genus levels in groups 1 and 3. CONCLUSIONS: Bacteroidetes to Firmicutes ratio and the abundance of Bifidobacterium were different according to the stool frequency, and specific bacteria were identified in the subjects with large and small numbers of defecation, respectively. These findings suggest that stool frequency might be associated with the richness and community composition of the gut microbiota.

Evaluation of fecal microbiomes associated with obesity in captive cynomolgus monkeys (Macaca fascicularis)

Microorganisms play important roles in obesity; however, the role of the gut microbiomes in obesity is controversial because of the inconsistent findings. This study investigated the gut microbiome communities in obese and lean groups of captive healthy cynomolgus monkeys reared under strict identical environmental conditions, including their diet. No significant differences in the relative abundance of Firmicutes, Bacteroidetes and Prevotella were observed between the obese and lean groups, but a significant difference in Spirochetes (p < 0.05) was noted. Microbial diversity and richness were similar, but highly variable results in microbial composition, diversity, and richness were observed in individuals, irrespective of their state of obesity. Distinct clustering between the groups was not observed by principal coordinate analysis using an unweighted pair group method. Higher sharedness values (95.81% ± 2.28% at the genus level, and 79.54% ± 5.88% at the species level) were identified among individual monkeys. This paper reports the association between the gut microbiome and obesity in captive non-human primate models reared under controlled environments. The relative proportion of Firmicutes and Bacteroidetes as well as the microbial diversity known to affect obesity were similar in the obese and lean groups of monkeys reared under identical conditions. Therefore, obesity-associated microbial changes reported previously appear to be associated directly with environmental factors, particularly diet, rather than obesity.

Effects of the Administration of Probiotics on Fecal Microbiota Diversity and Composition in Healthy Individuals

BACKGROUND/AIMS: Probiotics are expected to modify the composition of gut microbiota. We aimed to investigate the changes in the composition and diversity of gut microbiota by the administration of probiotics in healthy individuals. METHODS: Twelve healthy volunteers with age range of 30–42 years provided baseline fecal samples. Subsequently, they took commercially available probiotic capsules (a mixture for Bifidobacterium, Lactobacillus, and Enterococcus) for 4 weeks. Fecal samples were collected at 4 weeks of administration and 2 weeks after the stop of administration. Fecal microbiota was analyzed via 16S ribosomal RNA gene sequencing. RESULTS: The mean Shannon index was not significantly altered by the 4-week administration of probiotics (4.365 vs 4.556, P > 0.05). The proportion of Bacteroidetes, Actinobacteria, Firmicutes, and Proteobacteria was not significantly changed by the 4-week administration of probiotics. At the genus level, the proportions of Lactobacillus (2.138% vs 2.773%, P = 0.028) and Enterococcus (0.022% vs 2.758%, P = 0.004) significantly increased 4 weeks after the administration of probiotics, but reduced 2 weeks after the stop of administration (2.773% vs 3.292%, P = 0.064 and 2.758% vs 0.001%, P = 0.001). CONCLUSIONS: The diversity of fecal microbiota is not significantly affected by 4 weeks of probiotics administration. The proportion of fecal microbiota at the genus level is significantly altered by the administration of probiotics. However, this effect does not seem to last long, probably because of homeostasis or dietary influence.

Postoperative changes in intestinal microbiota and use of probiotics in roux-en-y gastric bypass and sleeve vertical gastrectomy: an integrative review / Mudanças na microbiota intestinal e uso de probióticos no pós-operatório de bypass gástrico em y-de-roux e gastrectomia vertical sleeve: uma revisão integrativa

ABSTRACT Introduction: Studies suggest that weight loss induced by bariatric surgery and the remission of some comorbidities may be related to changes in the microbiota profile of individuals undergoing this procedure. In addition, there is evidence that manipulation of the intestinal microbiota may prove to be a therapeutic approach against obesity and metabolic diseases. Objective: To verify the changes that occur in the intestinal microbiota of patients undergoing bariatric surgery, and the impact of the usage of probiotics in this population. Methods: Articles published between 2007 and 2017 were searched in Medline, Lilacs and Pubmed with the headings: bariatric surgery, microbiota, microbiome and probiotics, in Portuguese, English and Spanish. Of the 166 articles found, only those studies in adults subjected to either Roux-en-Y gastric bypass or sleeve vertical gastrectomy published in original articles were enrolled. In the end, five studies on the change of intestinal microbiota composition, four on the indirect effects of those changes and three on the probiotics administration on this population were enrolled and characterized. Conclusion: Bariatric surgery provides changes in intestinal microbiota, with a relative increase of the Bacteroidetes and Proteobacteria phyla and reduction of Firmicutes. This is possibly due to changes in the gastro-intestinal flux, coupled with a reduction in acidity, in addition to changes in eating habits. The usage of probiotics seems to reduce the gastro-intestinal symptoms in the post-surgery, favor the increase of vitamin B12 synthesis, as well as potentiate weight loss.

RESUMO Introdução: Estudos sugerem que a perda de peso induzida pela cirurgia bariátrica e a remissão de algumas comorbidades podem estar relacionadas às mudanças no perfil da microbiota dos indivíduos submetidos a este procedimento. Além disso, há indícios de que a manipulação da microbiota intestinal pode vir a ser abordagem terapêutica contra a obesidade e doenças metabólicas. Objetivo: Verificar as mudanças que ocorrem na microbiota intestinal de pacientes submetidos à cirurgia bariátrica, e o impacto do uso dos probióticos nessa população. Métodos: Foi realizada a busca de artigos publicados entre os anos de 2007 e 2017 nas bases de dados Medline, Lilacs e PubMed com os descritores: cirurgia bariátrica, microbiota, microbioma e probióticos em português, inglês e espanhol. Dos 166 artigos encontrados, foram selecionados apenas os estudos realizados em adultos, submetidos ao bypass gástrico em Y-de-Roux ou gastrectomia vertical sleeve publicados em artigos originais. Ao final, foram selecionados e categorizados cinco estudos sobre a mudança na composição da microbiota intestinal, quatro sobre os efeitos indiretos dessas mudanças e três sobre a administração de probióticos nessa população. Conclusão: A cirurgia bariátrica proporciona mudanças na microbiota intestinal com aumento relativo dos filos Bacteroidetes e Proteobactéria e redução de Firmicutes. Isso se deve, possivelmente, às alterações no trânsito gastrointestinal com redução da acidez intestinal além de modificação dos hábitos alimentares. O uso de probióticos parece reduzir os sintomas gastrointestinais no pós-operatório, favorecer o aumento de síntese de vitamina B12 e potencializar a perda de peso.

Calorie restriction and its impact on gut microbial composition and global metabolism / 医学前沿

Calorie restriction (CR) is a dietary regimen that reduces calorie intake without incurring malnutrition or a reduction in essential nutrients. It has long been recognized as a natural strategy for promoting health, extending longevity, and prevents the development of metabolic and age-related diseases. In the present review, we focus on the general effect of CR on gut microbiota composition and global metabolism. We also propose mechanisms for its beneficial effect. Results showed that probiotic and butyrate-producing microbes increased their relative abundance, whereas proinflammatory strains exhibited suppressed relative abundance following CR. Analyses of the gut microbial and host metabolisms revealed that most host microbial co-metabolites were changed due to CR. Examples of dramatic CR-induced changes in host metabolism included a decrease in the rate of lipid biosynthesis and an increase in the rates of fatty acid catabolism, β-oxidation, glycogenolysis, and gluconeogenesis. The observed phenotypes and the further verification of the direct link between gut microbiota and metabolome may benefit patients that are at risk for developing metabolic disease. Thus, improved gut microbiota composition and metabolome are potential biomarkers for determining the effectiveness of dietary interventions for age-related and metabolic diseases.

Influence of Chemical- and Natural-Based Lotions on Bacterial Communities in Human Forearm Skin

Purpose of this study was to evaluate the influence of a lotion on the bacterial community in the human forearm skin. The chemical- and natural-based lotions were applied on the left and right inner forearm skins, respectively, of 14 participants, who cleansed forearm skin using sterilized cotton swabs. The germs on cotton swabs were analyzed using libraries of PCR amplicons. The genetic diversity of the bacterial communities detected on the natural-based lotion-applied skin (NLS) was significantly higher than that of the bacterial communities on the chemical-based lotion-applied skin (CLS) in all participants, except two. The diversity was estimated based on operational taxonomic unit (OTU), Chao1, Shannon, and Simpson indices. Bacterial communities obtained from the CLS and NLS were phylogenetically separated into 5 and 3 monophyletic groups, respectively, based on lotion types. The taxonomic distribution of the bacterial communities, which were composed of 198 genera in 14 phyla in the CLS and NLS, respectively, was irregularly and biasedly separated into 2 groups based on the lotion types. Among the 14 phyla, Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were found to be relatively dominant, and 15 of the 198 genera, including Methylobacterium, Propionibacterium, Pseudomonas, Staphylococcus, Streptococcus, and Bacillus were relatively dominant (>0.5%). The taxonomic distribution of dominant bacterial communities from CLS and NLS was irregularly and biasedly separated without relation to the lotion types. In conclusion, the chemical- and natural-based lotions were responsible for changing or influencing the genetic diversity, phylogenetic separation, and taxonomic distribution of skin bacterial communities.

Comparisons of Gut Microbiota Among Healthy Control, Patients With Conventional Adenoma, Sessile Serrated Adenoma, and Colorectal Cancer

BACKGROUND: Studies on gut microbiota regarding colorectal carcinogenesis, including sessile serrated adenoma (SSA), have been scarce. The aim of this study is to investigate the role of mucosa-associated gut microbiota in the colorectal carcinogenesis. METHODS: We collected biopsy samples of normal rectal mucosa during colonoscopy from healthy control and patients with conventional adenoma, SSA, and colorectal cancer (CRC), respectively (n = 6). Pyrosequencing for 16S rRNA gene of bacteria was performed to compare gut microbiota. RESULTS: The most abundant phylum in total samples was Proteobacteria (55.6%), followed by Firmicutes (27.4%) and Bacteroidetes (11.6%). There was no significant difference in relative abundance of the phylum level among the four groups. Fusobacterium nucleatum, known to be frequently detected during colorectal carcinogenesis, was found in only one sample of patient with SSA. The rarefaction curves showed that the diversity of mucosal communities of patients with CRC is the lowest among the four groups and the diversity of mucosal communities of patients with SSA is higher than that of healthy control. Among the four groups, Shannon's and Simpson's index for diversity was the lowest and the highest in the patients with CRC, respectively; it did not reach statistical significance. The proportion of genus Pseudomonas was very high in the samples of patients with stage II–IV CRC compared with those with stage I CRC (59.3% vs. 0.3%, P = 0.064). CONCLUSIONS: Our study suggests no significant role of mucosa-associated gut microbiota in the colorectal carcinogenesis. Further study for many samples or using fecal material could be helpful.

Six-week Diet Correction for Body Weight Reduction and Its Subsequent Changes of Gut Microbiota: A Case Report

Impact of intestinal microbes on obesity and health is a new topic recently started to get attention. Comparing to the global concern and research trends, there are few research on the association between intestinal bacteria and life style disease in Korean. One voluntary case (female) was reported to show the change in gut microbiota and weight by diet intervention. She was overweight (BMI 23.2 kg/m2) and has abnormal liver function, and the causes of overweight were frequent drinking and meat consumption at the late evening hours. For 47 days, she was administered an improved diet on breakfast and dinner with reduction of meat consumption frequency by 50%. Alcohol consumption was reduced to once a week. As a result, she lost 3 kilograms of body weight. Her fecal sample was collected before and after the intervention, and gut microbiota change was compared using a high-throughput sequencing technique. After diet correction, the shift of gut microbiota was clearly observed with decreased proportion of Firmicutes (from 75.7% to 47.3% in total microbiota) but increased proportion of Bacteroidetes upto 47.7%. After incorporating the diet intervention, it is meaningful to confirm the changes in dominant gut microbiota and weight loss.

Gastric Microbiota and Gastric Cancer

The stomach had been recognized as an organ where many bacteria cannot survive due to the presence of gastric acid. However, a number of bacteria have been detected after the discovery of Helicobacter pylori with recent advances in nucleotide sequencing techniques and bioinformatics. These include Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria and Fusobacteria. Several animal studies suggested that the imbalance of gastric microbiotas could be associated with the development of gastric cancer. Changes in the composition of the gastric microbiota may increase the production of N-nitroso compounds, which is known to be a carcinogen. Further studies on the actual function and proteomics of gastric microbiota could be beneficial for prevention, early diagnosis, and new treatment strategies of gastric cancer.

Dynamics of Gut Microbiota According to the Delivery Mode in Healthy Korean Infants

Microbial colonization of the infant gut is unstable and shows a wide range of diversity between individuals. Gut microbiota play an important role in the development of the immune system, and an imbalance in these organisms can affect health, including an increased risk of allergic diseases. Microbial colonization of young infants is affected by the delivery mode at birth and the consequent alterations of gut microbiota in early life affect the development of allergic diseases. We investigated the effects of the delivery mode on the temporal dynamics of gut microbiota in healthy Korean infants. Fecal samples were collected at 1-3 days, 1 month, and 6 months after birth in six healthy infants. Microbiota were characterized by 16S rRNA shotgun sequencing. At the first and third days of life, infants born by vaginal delivery showed a higher richness and diversity of gut microbiota compared with those born by cesarean section. However, these differences disappeared with age. The Bacteroides genus and Bacteroidetes phylum were abundant in infants born by vaginal delivery, whereas Bacilli and Clostridium g4 were increased in infants born by cesarean section. The Firmicutes phylum and Bacteroides genus showed convergent dynamics with age. This study demonstrated the effect of delivery mode on the dynamics of gut microbiota profiles in healthy Korean infants.

Comparison of Gastric Microbiota Between Gastric Juice and Mucosa by Next Generation Sequencing Method

BACKGROUND: Not much is known about the role of gastric microbiota except for Helicobacter pylori in human health and disease. In this study, we aimed to detect human gastric microbiota in both gastric mucosa and gastric juice by barcoded 454-pyrosequencing of the 16S rRNA gene and to compare the results from mucosa and juice. METHODS: Gastric biopsies and stomach juices were collected from 4 subjects who underwent standard endoscopy at Seoul National University Bundang Hospital. Gastric microbiota of antral mucosa, corpus mucosa samples, and gastric fluids were analyzed by barcoded 454-pyrosequencing of the 16S rRNA gene. The analysis focused on bacteria, such as H. pylori and nitrosating or nitrate-reducing bacteria. RESULTS: Gastric fluid samples showed higher diversity compared to that of gastric mucosa samples. The mean of operational taxonomic units was higher in gastric fluid than in gastric mucosa. The samples of gastric fluid and gastric mucosa showed different composition of phyla. The composition of H. pylori and Proteobacteria was higher in mucosa samples compared to gastric fluid samples (H. pylori, 66.5% vs. 3.3%, P = 0.033; Proteobacteria, 75.4% vs. 26.3%, P = 0.041), while Actinobacteria, Bacteroidetes, and Firmicutes were proportioned relatively less in mucosa samples than gastric fluid. However there was no significant difference. (Actinobacteria, 3.5% vs. 20.2%, P = 0.312; Bacteroidetes, 6.0% vs. 14.8%, P = 0.329; Firmicutes, 12.8% vs. 33.4%, P = 0.246). CONCLUSIONS: Even though these samples were small, gastric mucosa could be more effective than gastric fluid in the detection of meaningful gastric microbiota by pyrosequencing.

Porphyromonas gingivalis Fim-A genotype distribution among Colombians / Distribución del genotipo Fim-A de Porphyromonas gingivalis entre la población colombiana

Introduction: Porphyromonas gingivalis is associated with periodontitis and exhibit a wide array of virulence factors, including fimbriae which is encoded by the FimA gene representing six known genotypes. Objetive: To identify FimA genotypes of P. gingivalis in subjects from Cali-Colombia, including the co-infection with Aggregatibacter actinomycetemcomitans, Treponema denticola, and Tannerella forsythia. Methods: Subgingival samples were collected from 151 people exhibiting diverse periodontal condition. The occurrence of P. gingivalis, FimA genotypes and other bacteria was determined by PCR. Results: Porphyromonas gingivalis was positive in 85 patients. Genotype FimA II was more prevalent without reach significant differences among study groups (54.3%), FimA IV was also prevalent in gingivitis (13.0%). A high correlation (p= 0.000) was found among P. gingivalis, T. denticola, and T. forsythia co-infection. The FimA II genotype correlated with concomitant detection of T. denticola and T. forsythia. Conclusions: Porphyromonas gingivalis was high even in the healthy group at the study population. A trend toward a greater frequency of FimA II genotype in patients with moderate and severe periodontitis was determined. The FimA II genotype was also associated with increased pocket depth, greater loss of attachment level, and patients co-infected with T. denticola and T. forsythia.

Introducción: Porphyromonas gingivalis es una bacteria asociada con la periodontitis. Expresa una amplia gama de factores de virulencia, incluyendo las fimbrias, las cuales están codificadas por el gen FimA que representa seis genotipos conocidos. Objetivo: Identificar los genotipos de FimA de P. gingivalis en pacientes de Cali - Colombia, incluyendo la co -infección con Aggregatibacter actinomycetemcomitans, Treponema denticola y Tannerella forsythia. Métodos: Se obtuvieron muestras subgingivales de 151 individuos con diferentes diagnósticos periodontales. La ocurrencia de P. gingivalis, los genotipos de FimA y otras bacterias se determinó por PCR. Resultados: Porphyromonas gingivalis fue positiva en 85 pacientes. El genotipo FimA II fue más prevalente, pero no hubo diferencias significativas entre los grupos de estudio (54.3%), FimA IV fue el más frecuente en la gingivitis (13.0%). Una alta correlación (p= 0.000) se encontró entre P. gingivalis , T. denticola y T. forsythia. El genotipo FimA II estuvo correlacionado con la detección de T. denticola y T. forsythia. Conclusiones: Porphyromonas gingivalis tuvo una alta frecuencia incluso en el grupo de individuos sanos. Se encontró una tendencia hacia una mayor frecuencia de FimA II en pacientes con periodontitis moderada y severa. El genotipo FimA II también se asoció con una mayor profundidad de la bolsa, una mayor pérdida de nivel de inserción, y con los pacientes en los que se detectó co-infección con T. denticola y T. forsythia.

Modulation of Gut Microbiota: Potential Mechanism of Diabetes Remission after Bariatric/Metabolic Surgery

Advances in genomic processing technology have been applied to the human microbiota and have provided the understanding of the effect of the microbiota in human health and disease. Especially, studies of the gut microbiota have revealed that changes in gut microbiota are related to obesity and the metabolic syndrome. With weight gain, there is an increase in the ratio of Firmicutes to Bacteroidetes. Certain bacteria have increased the ability to ferment dietary substrates, thereby increasing hosts' energy intake and weight gain. After transplantation of microbiota from lean donors, insulin sensitivity of recipients increased along with proliferation of butyrate-producing intestinal microbiota. One important finding after Roux-en-Y gastric bypass is the relative overabundance of Proteobacteria, which is different with the changes seen in weight loss without bypass surgery. Due to the change of microbiota's composition after bypass surgery, the gut environment is changed to unfavorable for energy absorption including decrease of polysaccharide fermentation. Therefore this change may contribute to the improvement of insulin resistance and loss of body weight. According to these results, modifying the gut microbiota through diet, probiotics, fecal transplants, and surgery might be included as therapeutic options for the diseases linked to imbalance in the microbiota.

Partial denture metal framework may harbor potentially pathogenic bacteria

PURPOSE: The aim of this study was to characterize and compare bacterial diversity on the removable partial denture (RPD) framework over time. MATERIALS AND METHODS: This descriptive pilot study included five women who were rehabilitated with free-end mandibular RPD. The biofilm on T-bar clasps were collected 1 week (t1) and 4 months (t2) after the RPD was inserted (t0). Bacterial 16S rDNA was extracted and PCR amplified. Amplicons were cloned; clones were submitted to cycle sequencing, and sequences were compared with GenBank (98% similarity). RESULTS: A total of 180 sequences with more than 499 bp were obtained. Two phylogenetic trees with 84 (t1) and 96 (t2) clones represented the bacteria biofilm at the RPD. About 93% of the obtained phylotypes fell into 25 known species for t1 and 17 for t2, which were grouped in 5 phyla: Firmicutes (t1=82%; t2=60%), Actinobacteria (t1=5%; t2=10%), Bacteroidetes (t1=2%; t2=6%), Proteobacteria (t1=10%; t2=15%) and Fusobacteria (t1=1%; t2=8%). The libraries also include 3 novel phylotypes for t1 and 11 for t2. Library t2 differs from t1 (P=.004); t1 is a subset of the t2 (P=.052). Periodontal pathogens, such as F. nucleatum, were more prevalent in t2. CONCLUSION: The biofilm composition of the RPD metal clasps changed along time after RPD wearing. The RPD framework may act as a reservoir for potentially pathogenic bacteria and the RPD wearers may benefit from regular follow-up visits and strategies on prosthesis-related oral health instructions.

Oral microbiota and host innate immune response in bisphosphonate-related osteonecrosis of the jaw / 国际口腔科学杂志·英文版

Bacterial biofilms have emerged as potential critical triggers in the pathogenesis of bisphosphonate (BP)-related osteonecrosis of the jaw (ONJ) or BRONJ. BRONJ lesions have shown to be heavily colonized by oral bacteria, most of these difficult to cultivate and presents many clinical challenges. The purpose of this study was to characterize the bacterial diversity in BRONJ lesions and to determine host immune response. We examined tissue specimens from three cohorts (n=30); patients with periodontal disease without a history of BP therapy (Control, n=10), patients with periodontal disease having history of BP therapy but without ONJ (BP, n=5) and patients with BRONJ (BRONJ, n=15). Denaturing gradient gel electrophoresis of polymerase chain reaction (PCR)-amplified 16S rRNA gene fragments revealed less bacterial diversity in BRONJ than BP and Control cohorts. Sequence analysis detected six phyla with predominant affiliation to Firmicutes in BRONJ (71.6%), BP (70.3%) and Control (59.1%). Significant differences (P<0.05) in genera were observed, between Control/BP, Control/BRONJ and BP/BRONJ cohorts. Enzyme-linked immunosorbent assay (ELISA) results indicated that the levels of myeloperoxidase were significantly lower, whereas interleukin-6 and tumor necrosis factor-alpha levels were moderately elevated in BRONJ patients as compared to Controls. PCR array showed significant changes in BRONJ patients with downregulation of host genes, such as nucleotide-binding oligomerization domain containing protein 2, and cathepsin G, the key modulators for antibacterial response and upregulation of secretory leukocyte protease inhibitor, proteinase 3 and conserved helix-loop-helix ubiquitous kinase. The results suggest that colonization of unique bacterial communities coupled with deficient innate immune response is likely to impact the pathogenesis of ONJ.