Effect of Moringa olifera leaves on growth and gut microbiota of Nile tilapia (Oreochromis niloticus) / Efeito das folhas da Moringa olifera no crescimento e na microbiota intestinal da tilápia do Nilo (Oreochromis niloticus)

Abstract The study was conducted to evaluate the effect of Moringa olifera on the growth and gut health of Tilapia (Oreochromis niloticus). The feed having 30% crude protein was prepared as an experimental diet with 4%, 8% and 10% M. olifera leaf supplementation, respectively. The control diet was devoid of M. olifera leaves. The 10 weeks feeding trial was carried out on 60 fish in aquaria. Fish was fed @ 3% of body weight twice a day. Diet with the high level of inclusion of M. olifera leaves significantly increased the growth rate, Survival Rate (SR), Specific Growth Rate (SGR) and Feed Conversion Efficiency (FCE) in all treatment groups compared to the control group. Similarly, Feed Conversion Ratio (FCR) gradually decreased and found highly-significant. To check the gut health of the Tilapia, random samples were selected and dissected. Nutrient agar was used as culture media to check the growth of bacteria. Pour Plate Method was used for viable colonies count by colony counter. Through staining method, the different bacteria such as Escherichia coli, Salmonella, Shigella and Pseudomonas aeruginosa were identify abundantly in the intestine of control diet fish but less number present in treatment diets groups. These results showed that M. olifera leaves up to 10% of dietary protein can be used for Nile tilapia for significant growth and healthy gut microbiota of fish.

Resumo O estudo foi conduzido para avaliar o efeito da Moringa olifera no crescimento e saúde intestinal da tilápia (Oreochromis niloticus). A ração com 30% de proteína bruta foi preparada como dieta experimental com 4%, 8% e 10% de suplementação de folhas de M. olifera, respectivamente. A dieta controle foi desprovida de folhas de M. olifera. O ensaio de alimentação de 10 semanas foi realizado em 60 peixes em aquários. O peixe pesava 3% do peso corporal duas vezes ao dia. A dieta com alto nível de inclusão de folhas de M. olifera aumentou significativamente a taxa de crescimento, taxa de sobrevivência (SR), taxa de crescimento de sobrevivência (SGR) e eficiência de conversão alimentar (FCE) em todos os grupos de tratamento em comparação com o grupo de controle. Da mesma forma, a taxa de conversão de alimentação (FCR) diminuiu gradualmente e foi considerada altamente significativa. Para verificar a saúde intestinal da tilápia, amostras aleatórias foram selecionadas e dissecadas. O ágar nutriente foi usado como meio de cultura para verificar o crescimento das bactérias. O método da placa de Verter foi usado para a contagem de colônias viáveis ​​por contador de colônias. Através do método de coloração, diferentes como Escherichia coli, Salmonella, Shigella e Pseudomonas aeruginosa foram identificados abundantemente no intestino de peixes da dieta controle, mas em menor número nos grupos de dieta de tratamento. Esses resultados mostraram que M. olifera deixa até 10% da proteína dietética e pode ser usado para tilápia do Nilo para um crescimento significativo e microbiota intestinal saudável de peixes.

Fecal microbiota profile of ovariohysterectomized cats submitted to estrogen replacement / Perfil da microbiota fecal de gatas ovariohisterectomizadas submetidas à reposição estrogênica

This study aimed to observe the effects of 17 ß-estradiol replacements on the fecal microbiota in spayed cats. Individual samples of fresh feces were collected and stored at -80° C. Sequencing of the V3/V4 regions of the 16S rRNA gene was used, and bioinformatic analysis was performed. Firmicutes/Bacteriodetes ratio was lower in the group receiving estrogen replacement compared to the SHAM group (P = 0,005). Jaccard index (P = 0.123) and Yue & Clayton index (P = 0.094) did not reveal alpha and beta diversity differences. The linear discriminant analysis effect size (LefSe) identified Firmicutes and MegasPhaera as the biomarkers for the SHAM group, and Burkholderiales, Betaproteobacteria, Sutterellaceae, Suterella, Proteobacteria, Proteobacteria unclassified and Collinsella for the group receiving estrogen replacement.(AU)

O objetivo deste estudo foi observar os efeitos da reposição de 17 ß-estradiol na microbiota fecal de gatas castradas. Amostras individuais de fezes frescas foram colhidas e armazenadas a -80°C. Foi realizado o sequenciamento das regiões V3/V4 do gene 16S rRNA e a análise bioinformática. A razão Firmicutes/Bacteriodetes foi menor no grupo que recebeu reposição estrogênica em comparação ao grupo SHAM (P = 0,005). O índice de Jaccard (P = 0,123) e o índice de Yue & Clayton (P = 0,094) não revelaram diferenças na alfa e beta diversidade. A análise discriminatória linear de tamanho do efeito (LefSe) identificou Firmicutes e Megasphaera como biomarcadores para o grupo SHAM, e Burkholderiales, Betaproteobacteria, Sutterellaceae, Suterella, Proteobacteria, Proteobacteria não classificada e Collinsella para o grupo que recebeu reposição estrogênica.(AU)

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

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 [...].

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 [...].

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.

Suplementação de glutamina na recuperação da flora intestinal / Glutamine supplementation in gut flora recovery / La suplementación con glutamina en la recuperación de la flora intestinal

Objetivo: evidenciar através de uma revisão integrativa os resultados clínicos atuais da suplementação de glutamina na melhora da saúde intestinal, através de sintomas e exames bioquímicos. Método: Revisão integrativa da literatura realizada no período de setembro de de 2021 nas bases de dados Pubmed e Scielo. Resultados: Foi realizado uma busca pelos descritores em saúde determinados e foram selecionadas 08 produções cientificas que atenderam os critérios de inclusão. Conclusão: Sugere-se novas pesquisas que elucidem as dosagens, efeitos colaterais e respostas terapêuticas da glutamina sobre parâmetros de saúde intestinal.

Objective: to evidence through an integrative review the current clinical results of glutamine supplementation in the improvement of intestinal health, through symptoms and biochemical tests. Method: Integrative review of the literature conducted in the period of September 2021 in the Pubmed and Scielo databases. Results: A search was performed for the defined health descriptors and 08 scientific productions were selected that met the inclusion criteria. Conclusion: Further research is suggested to elucidate the dosages, side effects and therapeutic responses of glutamine on intestinal health parameters.

Objetivo: evidenciar a través de una revisión integradora los resultados clínicos actuales de la suplementación con glutamina en la mejora de la salud intestinal, a través de síntomas y pruebas bioquímicas. Método: Revisión integradora de la literatura realizada en el periodo de septiembre de 2021 en las bases de datos Pubmed y Scielo. Resultados: Se realizó una búsqueda de los descriptores de salud definidos y se seleccionaron 08 producciones científicas que cumplieron con los criterios de inclusión. Conclusión: Se sugiere investigación adicional para dilucidar las dosis, los efectos secundarios y las respuestas terapéuticas de la glutamina en los parámetros de salud intestinal.

Association of nutritional status and diet with the gut microbiome in acutely ill hospitalised older adults / Associação do estado nutricional e da dieta com o microbioma intestinal em idosos hospitalizados agudamente enfermos

Introduction - Ageing is characterised by changes associated with an increased risk of developing chronic diseases and syndromes, of which malnutrition is one of the most common in older adults. The gut microbiota has an important role in the hosts health and is determined by several factors, including nutritional status and diet. Therefore, the gut microbiota may be associated with malnutrition and dietary intake in acutely ill older adults. Objectives - to identify- 1) the prevalence of malnutrition; 2) the association of malnutrition with the composition and metabolic potential of the gut microbiota and its impact on clinical outcomes; 3) the effect of habitual diet on the gut microbiota of acutely ill hospitalised older adults according to the nutritional status. Methods - a longitudinal analysis secondary to a prospective cohort was performed on 108 participants aged 65+ years old admitted to the hospital due to acute conditions. Clinical, demographic, nutritional, and anthropometric data and rectal swab samples were collected at admission and after 72 hours of hospitalisation. The food intake was estimated using the dietary history and the nutritional status diagnosed using the Global Leadership Initiative on Malnutrition (GLIM) criteria, preceded by the Mini-Nutritional Assessment Short-form (MNA-SF). Rectal swab samples were obtained to analyse the gut microbiota via 16S rRNA gene sequencing. The microbiota diversity, overall composition and differential abundances were calculated and compared between well-nourished and malnourished groups. Microbiome features potentially associated with malnutrition were selected by penalised models and confirmed by additive models. The prediction of clinical outcomes was investigated from the "malnourished microbiota" using decision trees. The dietary intake was explored through multivariate methods and investigated with the microbiota using tests of association and mediation analysis. Results - Malnourished patients (51%) had a different overall microbiota composition compared to well-nourished during hospitalisation (R= 0.079, p= 0.003). Severely malnourished (32.4%) showed a poorer diversity at admission (Shannon p= 0.012, Simpson p= 0.018) and after 72 hours (Shannon p= 0.023, Chao1 p= 0.008). Subdoligranulum, Lachnospiraceae NK4A136 group and Faecalibacterium prausnitzii, short-chain fatty acids producers, had significantly lower abundance and negative association with malnutrition, while Fusobacterium, Corynebacterium and Ruminococcaceae Incertae Sedis were highly increased and positively associated with malnutrition. Corynebacterium, Ruminococcaceae Incertae Sedis and the overall composition were important predictors of critical care in malnourished during hospitalisation. Malnourished patients had a significantly lower intake of plant protein, carbohydrates, dietary fibre, Fe, Mn, folate, campesterol, beta-sitosterol, and -linolenic acid, and higher intake of Vitamin D. There were no significant correlations of nutrients, foods or food groups with the microbiota structure, but for individual taxa and -diversity metrics. Mediation analysis revealed a significant indirect effect of the nutritional status on the differential abundance of several bacterial taxa partially mediated in a positive direction by dietary fibre, plant-based protein, onions, and olive oil, and negative by vitamin D. Conclusion - Malnourished patients had a significantly lower intake of key dietary compounds and substantial gut microbial disturbances during hospitalisation, pronounced in the severe stage. Some plant-based compounds might confer beneficial effects on the gut microbiota profile. A "malnourished microbiota" may be able to predict critical illness in hospitalised malnourished older patients. Bench-to-bedside investigations are necessary to confirm these findings.

Introdução - O envelhecimento é caracterizado por mudanças associadas a uma maior chance de desenvolver síndromes e doenças crônicas, das quais a desnutrição é uma das mais comuns em idosos. A microbiota intestinal tem um papel importante na saúde e é determinada por vários fatores, incluindo o estado nutricional e a dieta. Portanto, a microbiota intestinal pode estar associada à desnutrição e à ingestão alimentar em idosos gravemente doentes. Objetivos - identificar- 1) a prevalência de desnutrição; 2) a associação da desnutrição com a composição e o potencial metabólico da microbiota intestinal e seu impacto nos desfechos clínicos; 3) o efeito da dieta habitual sobre a microbiota intestinal de idosos hospitalizados com doenças agudas de acordo com o estado nutricional. Métodos - uma análise longitudinal secundária a uma coorte prospectiva foi realizada em 108 participantes com mais de 65 anos de idade admitidos no hospital devido a condições agudas. Foram coletados dados clínicos, demográficos, nutricionais, antropométricos e swab retal na admissão e após 72 horas de internação. O consumo alimentar foi estimado por meio do histórico alimentar e o estado nutricional foi diagnosticado pelos critérios da Iniciativa de Liderança Global sobre Desnutrição (GLIM), precedido da triagem pela Mini-Avaliação Nutricional versão curta (MAN-SF). Amostras de swab retal foram obtidas para analisar a microbiota intestinal via sequenciamento do gene 16S rRNA. A diversidade da microbiota, composição geral e a abundâncias diferenciais foram calculadas e comparadas entre os grupos bem nutrido e desnutrido. Variáveis do microbioma potencialmente associadas à desnutrição foram selecionadas por modelos penalizados e confirmadas por modelos aditivos. A predição para os desfechos clínicos foi investigada a partir da "microbiota desnutrida" utilizando árvores de decisão. O consumo alimentar foi explorado por meio de métodos multivariados e investigado com a microbiota por meio de testes de associação e análise de mediação. Resultados - Os pacientes desnutridos (51%) apresentaram composição geral da microbiota diferente comparado com os bem nutridos durante a hospitalização (R= 0,079, p= 0,003). Os sevemente desnutridos (32,4%) apresentaram menor diversidade na admissão (Shannon p= 0,012, Simpson p= 0,018) e após 72 horas (Shannon p = 0,023, Chao1 p= 0,008). Subdoligranulum, Lachnospiraceae NK4A136 e Faecalibacterium prausnitzii, produtores de ácidos graxos de cadeia curta, tiveram abundância significativamente menor e associação negativa com a desnutrição, enquanto Fusobacterium, Corynebacterium e Ruminococcaceae Incertae Sedis tiveram maior abundância e associação positiva. Corynebacterium, Ruminococcaceae Incertae Sedis e a composição geral foram importantes preditores de cuidados intensivos em desnutridos durante a hospitalização. Pacientes desnutridos tiveram ingestão significativamente menor de proteína vegetal, carboidratos, fibra dietética, Fe, Mn, folato, campesterol, beta-sitosterol e ácido -linolênico, e maior de vitamina D. Não houve correlações significativas entre nutrientes, alimentos ou grupos de alimentos com a estrutura da microbiota, mas sim para táxons individuais e métricas da -diversidade. A análise de medição revelou um efeito indireto significativo do estado nutricional sobre a abundância diferencial de vários táxons bacterianos parcialmente mediada em uma direção positiva pela fibra alimentar, proteína vegetal, cebola, e azeite de oliva, e negativa pela vitamina D. Conclusão - Pacientes desnutridos tiveram menor ingestão de compostos dietéticos chaves e distúrbios microbianos intestinais substanciais durante a hospitalização, pronunciados no estágio grave. Alguns compostos de origem vegetal podem conferir efeitos benéficos no perfil da microbiota intestinal. A "microbiota desnutrida" pode ser capaz de predizer doenças críticas em idosos desnutridos hospitalizados. Investigações pré-clínicas e translacionais são necessárias para confirmar esses achados.

Alteración de la microbiota intestinal y su relación con enfermedades gastrointestinales y hepatobiliares / Disruption of the intestinal microbiota and its relationship with gastrointestinal and hepatobiliary diseases

La interrupción de la simbiosis que existe entre el cuerpo humano y su microbioma puede resultar en una disbiosis, un desequilibrio en la interacción huésped-microbiota, que puede asociarse al desarrollo de diversas enfermedades como el síndrome de intestino irritable, hígado graso no alco-hólico, enfermedad hepática alcohólica y cirrosis, entre otras. En ciertas condiciones patológicas y por múltiples factores de riesgo, la capacidad de autorregulación del intestino se puede alterar, contribuyendo al incremento de la permeabilidad con inflamación intestinal crónica. El diagnóstico y el tratamiento, así como la relación entre la permeabilidad intestinal, la disbiosis y las patologías gastrointestinales y hepatobiliares, todavía no tienen estudios clínicos validados o con el soporte científico adecuado, por lo que se realiza una revisión de la literatura con la finalidad de aportar conceptos que puedan orientar con respecto a la importancia del estudio del microbioma humano en estas enfermedades.

Disruption of the symbiosis that exists between the human body and its microbiome can result in dys-biosis, an imbalance in the host-microbiota interaction, which may be associated with the develop-ment of various diseases such as irritable bowel syndrome, non-alcoholic fatty liver disease, alcoholic liver disease and cirrhosis, among others. In certain pathological conditions and due to multiple risk factors, the self-regulating capacity of the intestine may be lost, contributing to increased permeability with chronic intestinal inflammation. Its diagnosis and treatment as well as the relationship between intestinal permeability, dysbiosis and gastrointestinal and hepatobiliary pathologies have not been validated in clinical studies or have adequate scientific support, so a review of the literature is carried out in order to provide concepts that can guide with respect to the importance of the study of the human microbiome in these diseases

A relação do transtorno do espectro autista e a disbiose intestinal: uma revisão integrativa / The relationship between autism spectrum disorder and intestinal dysbiosis: an integrative review

Objetivos: evidenciar as relações e a existência do agravamento do Transtorno do Espectro Autista devido à disbiose intestinal. Métodos: revisão integrativa realizada segundo a pergunta norteadora: Existe comprovação científica entre a relação do TEA e disbiose intestinal que favoreça a melhora na prática clínica e indicações de possíveis respostas? Buscou-se por artigos publicados entre janeiro de 2016 e janeiro de 2021, nas bases de dados: PubMed, SciELO, LILACS, GOOGLE ACADÊMICO. Foram utilizados os descritores (DeCS): "Transtorno do Espectro Autista"; "Microbiota gastrointestinal"; "Disbiose", associados pelo operador booleano "E". Foram incluidos artigos de revisões bibliográficas, completos, originais, limitados aos idiomas inglês e português brasileiro, publicados nos últimos cinco anos, e que, após leitura do resumo, estivessem dentro do escopo da revisão. Resultados: Foram identificados 52 manuscritos e, após aplicação dos critérios de inclusão e exclusão, foram considerados 11 artigos que evidenciam o agravamento do TEA por fatores intrínsecos à microbiota intestinal. Conclusão: existe importante influência causal do eixo bidirecional cérebro-intestino-microbiota na etiologia e exacerbaçao das manifestações clínicas do Transtorno do Espectro Autista devido à disbiose intestinal e aos fatores gastrointestinais de origem idiopática.

Objectives: to highlight the relationships and the existence of Autistic Spectrum Disorder aggravation due to intestinal dysbiosis. Methods: integrative review conducted according to the guiding question: Is there scientific evidence of the relationship between ASD and intestinal dysbiosis that favors improvement in clinical practice and indications of possible answers? We searched for articles published between January 2016 and January 2021 in databases: PubMed, SciELO, LILACS, and GOOGLE ACADEMIC. The following descriptors (DeCS) were used: "Autistic Spectrum Disorder"; "Gastrointestinal microbiota"; "Dysbiosis", associated with the Boolean operator "AND". We included literature review articles, complete, original, limited to English and Brazilian Portuguese languages, published in the last five years, and which, after reading the abstract, were within the scope of the review. Results: 52 manuscripts were identified, and after applying the inclusion and exclusion criteria, 11 articles were considered that show the worsening of ASD due to factors intrinsic to the intestinal microbiota. Conclusion: there is an important causal influence of the bidirectional brain-gut-microbiota axis in the etiology and exacerbation of clinical manifestations of Autism Spectrum Disorder due to intestinal dysbiosis and gastrointestinal factors of idiopathic origin.

Qual é a influência da microbiota na obesidade e em seu quadro inflamatório? / What is the influence of microbiota on obesity and its inflammatory involvement?

A obesidade tem causa multifatorial que atinge atualmente mais da metade da população brasileira. Mais recentemente, a microbiota intestinal foi considerada um fator que contribui para essa condição. Os objetivos deste estudo foram revisar a influência da microbiota intestinal na obesidade e no processo inflamatório, e analisar os efeitos da utilização dos pré e probióticos. Foi realizada revisão sistemática sobre o assunto. Dos mais de 27.000 artigos, apenas 16 respeitaram os critérios de inclusão. Em conclusão, o desequilíbrio da microbiota aparece como fator favorável ao desenvolvimento da obesidade e do quadro inflamatório decorrente dela. Tanto o uso de prebióticos quanto probióticos são recursos válidos no tratamento da obesidade, porém os primeiros parecem proporcionar melhor qualidade de vida.

Obesity has a multifactorial etiological condition that involves more than half of the Brazilian population. More recently, the intestinal microbiota was considered a factor that contributes to this condition. The aims of this study were to review the intestinal microbiota influence in the obesity and in the inflammatory response, and to analyze the effects of using prebiotic and probiotic medications. A systematic review was firstly done. More than 27,000 articles were found, but only 16 contained the proper criteria. In conclusion, the microbiota imbalance seems to increase the obesity development and its inflammatory aspects. Both the use of pre and probiotics are good options in the obesity treatment, though the first ones seem to enhance bettere quality of life.

Microbiota intestinal y modulación del tejido adiposo en la patogénesis de la obesidad / Gut microbiota and modulation of adipose tissue in the pathogenesis of obesity

Las investigaciones realizadas durante el último siglo relacionadas con la descripción de la Microbiota Intestinal (MI) sugieren una relación concreta entre su composición y la salud del huésped. Su desregulación denominada disbiosis intestinal ha sido asociada a distintos tipos de enfermedades gastrointestinales, metabólicas, oncológicas e incluso psiquiátricas. Destacan numerosos reportes que han informado la condición de disbiosis en la obesidad, tanto en modelos animales como humanos de distintos grupos etarios y regiones del mundo. A su vez, la composición del microbioma también ha logrado asociarse a las diferentes comorbilidades de la obesidad, postulando que la MI posee influencia en la disfunción del tejido adiposo (TA), entendiendo que corresponde al principal modulador de la patogénesis de la obesidad. Sin embargo, aún no es posible establecer una explicación mecanicista plausible. Actualmente, la utilización de tecnologías multiómicas, junto con la evaluación de variables fisiológicas, nos podrían proporcionar una mejor comprensión a la incógnita planteada. Frente a esto, el presente trabajo tiene como objetivo revisar los últimos avances en la comprensión de la influencia de la microbiota intestinal en el TA y su contribución a los mecanismos relacionados con la patogénesis de la obesidad. Entre los principales mecanismos identificados, la evidencia reporta nexos fisiológicos entre la composición de la MI y la modulación de inflamación, permeabilidad intestinal y adipogénesis. Las vías implicadas derivan de la influencia de la disbiosis intestinal en el accionar de ácidos grasos de cadena corta, claudinas, macrófagos, oligosacáridos, entre otros. Los mecanismos implicados, principalmente estudiados en modelos animales, deberían ser considerados para su evaluación en próximos estudios longitudinales y experimentales en humanos con el fin de obtener una mayor comprensión sobre la implicancia de cada mecanismo en la patogenia global de la obesidad(AU)

The investigations carried out during the last century related to the description of the Gut Microbiota (GM) suggest a concrete relationship between its composition and the health of the host. Its deregulation called intestinal dysbiosis has been associated with different types of gastrointestinal, metabolic, oncological and even psychiatric diseases. Numerous reports that have described the condition of dysbiosis in obesity stand out, both in animal and human models of different age groups and regions of the world. In turn, the composition of the microbiome has also been associated with the different comorbidities of obesity, postulating that MI has an influence on adipose tissue (AT) dysfunction, understanding that it corresponds to the main modulator of the pathogenesis of obesity. However, it is not yet possible to establish a plausible mechanistic explanation. Currently, the use of multi-omics technologies, together with the evaluation of physiological variables, could provide us with a better understanding of the question raised. In view of this, this review aims to review the latest advances in understanding the influence of the intestinal microbiota on AT and its contribution to the mechanisms related to the pathogenesis of obesity. Among the main mechanisms identified, the evidence reports physiological links between the composition of GM and the modulation of inflammation, intestinal permeability and adipogenesis. The pathways involved derive from the influence of intestinal dysbiosis on the action of short-chain fatty acids, claudins, macrophages, oligosaccharides, among others. The mechanisms involved, mainly studied in animal models, should be considered for evaluation in future longitudinal and experimental studies in humans in order to obtain a better understanding of the implication of each mechanism in the global pathogenesis of obesity(AU)

Microbiota, probióticos y el comportamiento humano / Microbiota, probiotics, and the human behavior / Microbiota, probióticos e comportamento humano

En la actualidad, ha cobrado una gran importancia la relación que la microbiota intestinal mantiene con varios órganos y sistemas del cuerpo humano. Particularmente importante, son las relaciones de la microbiota con el Sistema Nervioso Central, el comportamiento y el desarrollo y tratamiento de varias enfermedades. La relación existente entre la microbiota intestinal y el cerebro se produce gracias a la actividad de estímulos neuroendocrinos y neuroinmunes que pueden actuar de forma bilateral, llegando incluso a generar modificaciones en el comportamiento del ser humano. Del mismo modo, a través de la realización de estudios clínicos y paraclínicos, se ha conseguido demostrar la asociación entre el eje microbiota-intestino-cerebro y trastornos neurológicos como la enfermedad de Parkinson o el trastorno depresivo. El objetivo del presente artículo es realizar un análisis de los principales estudios identificados en relación a la función del eje microbiota-intestino-cerebro (MIC) así como identificar la nueva evidencia acerca del uso de probióticos en el tratamiento coadyuvante de varios trastornos neuro-psiquiátricos. Se realizó una búsqueda sistemática de la bibliografía utilizando palabras claves y términos MeSH y se presentó en formato de discusión de acuerdo a los subtemas: eje microbiota-intestino-cerebro, mecanismos de acción, microbiota y su relación con el comportamiento y regulación sobre probióticos. Se concluyó que existe evidencia que demuestra la relación entre el eje microbiota-intestino-cerebro y varios trastornos neuropsiquiátricos en el ser humano. Además, que la administración de probióticos puede modificar el eje MIC y pueden constituir una alternativa de terapia coadyuvante en estos trastornos del comportamiento.

Nowadays, the relationship that the intestinal microbiota maintains with various organs and systems of the human body has gained more importance. Especially relevant are the relationships of the microbiota with the Central Nervous System, behavior, and the development and treatment of various diseases. The relationship between the intestinal microbiota and the brain is a product of neuroendocrine and neuroimmune stimuli that can act bilaterally, even generating changes in human behavior. Moreover, clinical and paraclinical studies have demonstrated the association between the microbiota-gut-brain axis and neurological disorders such as Parkinson's disease or depressive disorder. The objective of this article is to carry out an analysis of the studies concerning the function of the microbiota-gut-brain (MGB) axis, as well as to identify new evidence about the use of probiotics in the adjunctive treatment of several neuropsychiatric disorders. A systematic search of the bibliography was carried out using keywords and MeSH terms and presented in a discussion format according to the subtopics: microbiota-gut-brain axis, mechanisms of action, microbiota, and its relationship with behavior and regulation on probiotics. The conclusion was that the evidence demonstrates the relationship between the microbiota-gut-brain axis and several neuropsychiatric disorders in humans. In addition, the administration of probiotics can modify the MGB axis and constitute an alternative for adjuvant therapy in these behavioral disorders.

A relação da microbiota intestinal com vários órgãos e sistemas do corpo humano tem se tornado cada vez mais importante. Particularmente importantes são as relações da microbiota com o sistema nervoso central, o comportamento e o desenvolvimento e tratamento de várias doenças. A relação entre a microbiota intestinal e o cérebro ocorre através da atividade de estímulos neuroendócrinos e neuroimunes que podem agir bilateralmente, levando até mesmo a mudanças no comportamento humano. Da mesma forma, estudos clínicos e paraclínicos demonstraram a associação entre o eixo microbiota-cérebro-cérebro e desordens neurológicas, como a doença de Parkinson ou desordem depressiva. O objetivo deste artigo é rever os principais estudos identificados em relação ao papel do eixo microbiota-cérebro-cérebro (MIC) e identificar novas evidências sobre o uso de probióticos no tratamento adjuvante de vários distúrbios neuropsiquiátricos. Uma pesquisa sistemática da literatura foi realizada usando palavras-chave e termos MeSH e apresentada em formato de discussão de acordo com os subtemas: eixo microbiota-cérebro-cérebro, mecanismos de ação, microbiota e sua relação com o comportamento e regulamentação sobre probióticos. Concluiu-se que há evidência de uma relação entre o eixo microbiota-cérebro-cérebro e vários distúrbios neuropsiquiátricos em humanos. Além disso, a administração de probióticos pode modificar o eixo MIC e pode constituir uma terapia adjuvante alternativa nestes distúrbios comportamentais.

Metabolitos bioactivos generados por la disbiosis intestinal y sus implicaciones fisiopatológicas en la enfermedad cardiovascular / Bioactive metabolites generated by intestinal dysbiosis and their pathophysiological implications in cardiovascular disease

Introducción: La disbiosis conocida como la alteración de la relación simbiótica entre la microbiota intestinal y el huésped están implicados en la patogenia de la enfermedad cardiovascular aterosclerótica. Objetivo: Realizar una revisión documental sobre los mecanismos fisiopatológicos que relacionan los metabolitos bioactivos generados por la disbiosis intestinal con el desarrollo y progresión de la enfermedad cardiovascular aterosclerótica. Métodos: Se utilizó el motor de búsqueda Google Académico y se consultaron artículos de libre acceso en las bases de datos Pubmed, SciELO, Lilacs, Cumed y Hinari desde septiembre 2020 hasta el mes de marzo 2021. Las palabras clave utilizadas para esta revisión fueron:microbioma, microbiota intestinal, disbiosis, aterosclerosis, enfermedad cardiovascular y sus equivalentes en inglés, según el descriptor de Ciencias de la Salud (DeCS). Se consideraron artículos originales, de revisión, revisiones sistemáticas y metaanálisis posteriores al año 2015. Se revisaron un total de 73 artículos. Desarrollo: Las relaciones fisiopatológicas entre la disbiosis intestinal y las enfermedades cardiovasculares son complejas, ya que se influyen mutuamente a través de los sus toxinas endógenas (metabolitos bioactivos), el sistema circulatorio, las respuestas inmunitarias y los cambios metabólicos. Las investigaciones futuras deberían centrarse en dilucidar los actores moleculares subyacentes e identificar si las vías que interconectan la disbiosis intestinal con la ECA son causales, correlacionales o consecuentes. Conclusiones: La evidencia acumulada sostiene que la disbiosis de la microbiota intestinal está involucrada en la síntesis de metabolitos proaterogénicos los cuales modulan los mecanismos implicados en la fisiopatología de la ECA(AU)

Introduction: Dysbiosis is known as the alteration of the symbiotic relationship between the intestinal microbiota and the host is involved in the pathogenesis of atherosclerotic cardiovascular disease. Objective: To carry out a documentary review on the pathophysiological mechanisms that relate the bioactive metabolites generated by intestinal dysbiosis with the development and progression of atherosclerotic cardiovascular disease. Methods: The Google Scholar search engine was used and free access articles were consulted in Pubmed, SciELO, Lilacs, Cumed and Hinari databases from September 2020 to March 2021. The keywords used for this review were microbiome, gut microbiota, dysbiosis, atherosclerosis, cardiovascular disease and their English equivalents, according to the Health Sciences (DeCS) descriptor. Original articles, review articles, systematic reviews and meta-analyses after 2015 were considered. A total of 73 articles were reviewed. Findings: The pathophysiological relationships between intestinal dysbiosis and cardiovascular diseases are complex, since they influence each other through their endogenous toxins (bioactive metabolites), the circulatory system, immune responses and metabolic changes. Future research should focus on elucidating the underlying molecular players and on identifying whether the pathways that interconnect gut dysbiosis with ACE are causal, correlational, or consequential. Conclusions: The accumulated evidence supports that the dysbiosis of the intestinal microbiota is involved in the synthesis of proatherogenic metabolites which modulate the mechanisms involved in the pathophysiology of ACE(AU)

Gut microbiome in neuropsychiatric disorders / O microbioma intestinal nas doenças neuropsiquiátricas

ABSTRACT Background: Neuropsychiatric disorders are a significant cause of death and disability worldwide. The mechanisms underlying these disorders include a constellation of structural, infectious, immunological, metabolic, and genetic etiologies. Advances in next-generation sequencing techniques have demonstrated that the composition of the enteric microbiome is dynamic and plays a pivotal role in host homeostasis and several diseases. The enteric microbiome acts as a key mediator in neuronal signaling via metabolic, neuroimmune, and neuroendocrine pathways. Objective: In this review, we aim to present and discuss the most current knowledge regarding the putative influence of the gut microbiome in neuropsychiatric disorders. Methods: We examined some of the preclinical and clinical evidence and therapeutic strategies associated with the manipulation of the gut microbiome. Results: targeted taxa were described and grouped from major studies to each disease. Conclusions: Understanding the complexity of these ecological interactions and their association with susceptibility and progression of acute and chronic disorders could lead to novel diagnostic biomarkers based on molecular targets. Moreover, research on the microbiome can also improve some emerging treatment choices, such as fecal transplantation, personalized probiotics, and dietary interventions, which could be used to reduce the impact of specific neuropsychiatric disorders. We expect that this knowledge will help physicians caring for patients with neuropsychiatric disorders.

RESUMO Antecedentes: Os transtornos neuropsiquiátricos são uma importante causa de morte e invalidez no mundo. Os mecanismos subjacentes a esses transtornos incluem uma constelação de etiologias estruturais, infecciosas, imunológicas, metabólicas e genéticas. Avanços nas técnicas de sequenciamento do DNA têm demonstrado que a composição do microbioma entérico é dinâmica e desempenha um papel fundamental não apenas na homeostase do hospedeiro, mas também em várias doenças. O microbioma entérico atua como mediador na sinalização das vias metabólica, neuroimune e neuroendócrina. Objetivo: Apresentar os estudos mais recentes sobre a possível influência do microbioma intestinal nas diversas doenças neuropsiquiátricas e discutir tanto os resultados quanto a eficácia dos tratamentos que envolvem a manipulação do microbioma intestinal. Métodos: foram examinadas algumas das evidências pré-clínicas e clínicas e estratégias terapêuticas associadas à manipulação do microbioma intestinal. Resultados: os táxons-alvo foram descritos e agrupados a partir dos principais estudos para cada doença. Conclusões: Entender a fundo a complexidade das interações ecológicas no intestino e sua associação com a suscetibilidade a certas doenças agudas e crônicas pode levar ao desenvolvimento de novos biomarcadores diagnósticos com base em alvos moleculares. Além disso, o estudo do microbioma intestinal pode auxiliar na otimização de tratamentos não farmacológicos emergentes, tais como o transplante de microbiota fecal, o uso de probióticos e intervenções nutricionais personalizadas. Dessa forma, terapias alternativas poderiam ser usadas para reduzir o impacto dos transtornos neuropsiquiátricos na saúde pública. Esperamos que esse conhecimento seja útil para médicos que cuidam de pacientes com diversos transtornos neuropsiquiátricos.

Modulación inmune en enfermedades alérgicas asociada al uso de Bifidobacterium en menores de cinco años / Immune modulation in allergic diseases associated with the use of Bifidobacterium in children under five years of age

La incidencia de enfermedades alérgicas en la infancia va en aumento, y se ha convertido en una de las principales consultas. Una posible causa es la disbiosis del microbioma intestinal, relacionada con estados inflamatorios aumentados. Debido a la necesidad de mejorar la calidad de vida, y el impacto en lo económico y en lo educativo, surgen los probióticos como tratamiento adyuvante, por lo que se pretende determinar la asociación del uso de Bifidobacterium en menores de 5 años con la modulación de la respuesta inmune en enfermedades alérgicas. El microbioma intestinal inicia su desarrollo y maduración desde la gestación, continúa en el nacimiento y termina hasta los 3 años, influenciado por factores maternos, neonatales y ambientales. La disbiosis intestinal generada por estos factores reduce la proporción de bifidobacterias, lo cual se relaciona con estados proinflamatorios. En consecuencia, estudios del uso de Bifidobacterium en niños con enfermedades alérgicas ha evidenciado mejora de síntomas y calidad de vida. Los probióticos favorecen un microbioma intestinal saludable, asociado a un estado antiinflamatorio, debido a la regulación en el balance celular Th1/Th2/T reguladoras y células asesinas naturales. Esta modulación en la respuesta inmune permite mejor control de síntomas, calidad de vida y menor incidencia de enfermedades alérgicas en la infancia

The incidence of allergic diseases in childhood is increasing, and has become one of the main queries. One possible cause is dysbiosis of the gut microbiome, related to increased inflammatory states. Due to the need to improve the quality of life, and the economic and educational impact, probiotics emerge as adjuvant treatment, so it is intended to determine the association of the use of Bifidobacterium in children under 5 years with the modulation of the immune response in allergic diseases. The intestinal microbiome begins its development and maturation from gestation, continues at birth and ends up to 3 years, influenced by maternal, neonatal and environmental factors. The intestinal dysbiosis generated by these factors reduces the proportion of bifidobacteria, which is related to proinflammatory states. Consequently, studies of the use of Bifidobacterium in children with allergic diseases have shown improvement in symptoms and quality of life. Probiotics favor a healthy intestinal microbiome, associated with an anti-inflammatory state, due to the regulation of the regulatory Th1/Th2/T cell balance and natural killer cells. This modulation in the immune response allows better control of symptoms, quality of life and lower incidence of allergic diseases in childhood

Role and change of the gut microbiota after bariatric surgery / 中华胃肠外科杂志

Gut microbiota have been validated to play a pivotal role in metabolic regulation. As the most effective treatment for obesity and related comorbidities, bariatric surgery has been shown to result in significant alterations to the gut microbiota. Literature have recently suggested temporal and spatial features of alterations to the intestinal bacteria following bariatric surgery, which is possibly attributed to the gut adaptation to the surgical modification on the gastrointestinal tract. More importantly, the gut microbiota have been appreciated as a critical contributor to the metabolic improvements following bariatric surgery. Although not fully elucidated, the underlying mechanisms are associated with the molecular pathways mediating the crosstalk between gut microbiota and host . On the other hand, change of the gut microbiota has been found to be related to the prognosis of patients receiving bariatric surgery. Some studies even point out negative effects of the gut microbiota on certain surgical complications . In this review, we summarize the characteristics of alterations to the gut microbiota following bariatric surgery as well as its relevant impacts to better understand the role of gut microbiota in bariatric surgery.

Using 16S rDNA Sequencing Technology to Preliminarily Analyze Intestinal Flora in Children with Mycoplasma pneumoniae Pneumonia / 生物医学与环境科学（英文）

Objective@#We investigated changes in the intestinal flora of children with Mycoplasma pneumoniae pneumonia (MPP).@*Methods@#Between September 2019 and November 2019, stool samples from 14 children with MPP from The Fourth Hospital of Baotou city, Inner Mongolia Autonomous Region, were collected and divided into general treatment (AF) and probiotic (AFY) groups, according to the treatment of "combined Bifidobacterium, Lactobacillus, Enterococcus, and Bacillus cereus tablets live". High-throughput 16S rDNA sequencing was used to identify intestinal flora.@*Results@#Intestinal flora abundance and diversity in children with MPP were decreased. Both Shannon and Simpson indices were lower in the AF group when compared with healthy controls ( P < 0.05). When compared with healthy controls, the proportion of Enterorhabdus was lower in the AF group, while the proportion of Lachnoclostridium was higher ( P < 0.05). The proportion of Bifidobacteria and Akkermansia was lower in the AFY group but Enterococcus, Lachnoclostridium, Roseburia, and Erysipelatoclostridium proportions were higher. The proportion of Escherichia coli- Shigella in the AFY group after treatment was decreased ( P < 0.05).@*Conclusions@#The intestinal flora of children with MPP is disturbed, manifested as decreased abundance and diversity, and decreased Bifidobacteria. Our probiotic mixture partly improved intestinal flora disorders.

Research progress on the regulation of mammalian energy metabolism by the circadian clock system and gut microbiota / 生理学报

The mammalian internal circadian clock system has been evolved to adapt to the diurnal changes in the internal and external environment of the organism to regulate diverse physiological functions, such as the sleep-wake cycle and feeding rhythm, thereby coordinating the rhythmic changes of energy demand and nutrition supply in each diurnal cycle. The circadian clock regulates glucose metabolism, lipid metabolism, and hormones secretion in diverse tissues and organs, including the liver, skeletal muscle, pancreas, heart, and vessels. As a special "organ" of the host, the gut microbiota, together with the intestinal microenvironment (tissues, cells, and metabolites) in a co-evolutionary process, constitutes a micro-ecosystem and plays an important role in the process of nutrient digestion and absorption in the intestine of the host. In recent years, accumulating evidence indicates that the compositions, quantities, colonization, and functional activities of the gut microbiota exhibit significant circadian variations, which are closely related to the changes of various physiological functions under the regulation of host circadian clock system. In addition, several studies have shown that the gut microbiota can produce many important metabolites such as the short-chain fatty acids through the degradation of indigestive dietary fibers. A portion of gut microbiota-derived metabolites can regulate the circadian clock system and metabolism of the host. This article mainly discusses the interaction between the host circadian clock system and the gut microbiota, and highlights its influence on energy metabolism of the host, providing a novel clues and thought for the prevention and treatment of metabolic diseases.

Changes in the structure of intestinal mucosal flora in colorectal cancer patients / 南方医科大学学报

OBJECTIVE@#To investigate the changes in bacterial flora in fecal samples, at the tumor loci and in adjacent mucosa in patients with colorectal cancer (CRC).@*METHODS@#We collected fecal samples from 13 patients with CRC and 20 healthy individuals and tumor and adjacent mucosa samples from 6 CRC patients. The differences in bacterial composition between the fecal and mucosa samples were analyzed with 16S rDNA sequencing and bioinformatics methods. We also detected the total number of bacteria in the feces using flow cytometry, isolated and identified the microorganisms in the fecal and mucosa samples using common bacterial culture media. We further tested the effects of 7 isolated bacterial strains on apoptosis of 3 CRC cell lines using lactate dehydrogenase detection kit.@*RESULTS@#The bacterial α-diversity in the feces of healthy individuals and in adjacent mucosa of CRC patients was significantly higher than that in the feces and tumor mucosa in CRC patients (P < 0.05). Lactobacillaceae is a specific bacteria in the feces, while Escherichia, Enterococcus, and Fusobacterium are specific bacteria in tumor mucosa of CRC patients as compared with healthy individuals. Cell experiment with3 CRC cell lines showed that Bacteroides fragilis isolated from the tumor mucosa of CRC patients produced significant inhibitory effects on cell proliferation (P < 0.0001), while the isolated strain Fusobacterium nucleatum obviously promoted the proliferation of the cell lines (P < 0.001).@*CONCLUSION@#The bacterial flora in the feces, tumor mucosa and adjacent mucosa of CRC patients is significantly different from that in the feces of healthy individuals, and the fecal flora of CRC patients can not represent the specific flora of the tumor mucosa. Inhibition of F. nucleatum colonization in the tumor mucosa and promoting B. fragilis colonization may prove beneficial for CRC treatment.

Investigating the effects of Modified Sijunzi Decoction on the diversity of intestinal microflora of severe scald rabbits based on 16S ribosomal RNA high-throughput sequencing / 中华烧伤杂志

Objective: To investigate the effects of Modified Sijunzi Decoction on the diversity of intestinal microflora of in severe scald rabbits based on 16S ribosomal RNA (16S rRNA) high-throughput sequencing. Methods: The experimental research method was adopted. Ninety Japanese big-ear rabbits regardless gender, aged 6 to 8 months, were randomly divided into normal control group, scald alone group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group, with 18 rabbits in each group. The rabbits in normal control group were free to eat and drink, and the rabbits in scald alone group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group were intragastrically administered normal saline, 0.2 g/mL Modified Sijunzi Decoction, 1.0 g/mL Modified Sijunzi Decoction, and 5.0 g/mL Modified Sijunzi Decoction, respectively for 7 days after sustaining full-thickness scalding of 30% total body surface area. On the 1st, 3rd, and 7th day after grouping, the levels of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-10 in ileal mucosa tissue of rabbits in each group were determined by enzyme-linked immunosorbent assay, and the number of samples in each group at each time point was 6. According to the above experimental results, another 9 rabbits were selected and divided into normal control group, scald alone group and scald+medium-dose group, with 3 rabbits in each group. The grouping and treatment methods of rabbits in each group were the same as before. On the 7th day after grouping, the V3, V4 region of 16S rRNA of ileum mucosa of rabbits in three groups were sequenced by high-throughput sequencing technology. The number of quality bacteria was counted by QIME software. The classifications of phylum, class, order, family and genus of microflora were analyzed by RDP Classifier software. The α diversity (Ace, Chao1, Simpson, and Shannon indexes) and β diversity were analyzed by Illumina MiSeq sequencing technology, and the number of experiment samples in each group was 3. Data were statistically analyzed with analysis for variance of factorial design, SNK test, and Bonferroni correction. Results: Compared with that in normal control group, the levels of TNF-α of ileal mucosa tissue of rabbits in scald alone group, scald+low-dose group, and scald+high-dose group on the 1st, 3rd, and 7th day after grouping and scald+medium-dose group on the 1st and 3rd day after grouping were all significantly increased (P<0.01), the levels of IL-1β in ileal mucosa tissue of rabbits in scald alone group, scald+low-dose group, scald+medium-dose group and scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly increased (P<0.05 or P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald alone group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly decreased (P<0.01). Compared with that in scald alone group, the levels of TNF-α in ileal mucosa tissue of rabbits in scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 3rd and 7th day after grouping, and scald+medium-dose group on the 1st day after grouping were all significantly decreased (P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 3rd and 7th day after grouping and scald+medium-dose group on the 1st day after grouping were all significantly decreased (P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald+low-dose group on the 7th day after grouping and scald+medium-dose group on the 1st, 3rd, and 7th day after grouping and scald+high-dose group on the 3rd and 7th day after grouping were all significantly increased (P<0.05 or P<0.01). Compared with that in scald+low-dose group, the levels of TNF-α in ileal mucosa tissue of rabbits in medium-dose scald alone group on the 1st, 3rd, and 7th day after grouping and in high-dose scald alone group on the 3rd and 7th day after grouping were significantly decreased (P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in medium-dose scald alone group on the 1st, 3rd, and 7th day after grouping and in high-dose scald alone group on the 3rd and 7th day after grouping were all significantly decreased (P<0.05 or P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald+medium-dose group on the 1st, 3rd, and 7th day after grouping and in scald+high-dose group on the 7th day after grouping were all significantly increased (P<0.05 or P<0.01). Compared with that in scald medium-dose group, the levels of TNF-α in ileal mucosa tissue of rabbits in scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly increased (P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald+high-dose group on the 1st, 3rd, and 7th day after grouping were all significantly decreased (P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in scald+high-dose group on the 7th day after grouping was significantly decreased (P<0.01). Compared with that on the 1st day after grouping, the levels of TNF-α in ileal mucosa tissue of rabbits in scald alone group on the 3rd and 7th day after grouping and in normal control group on the 3rd day after grouping were all significantly increased (P<0.05 or P<0.01), and the levels of IL-1β in ileal mucosa tissue of rabbits in scald alone group both on the 3rd and 7th day after grouping were significantly increased (P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in both scald+low-dose group and scald+high-dose group on the 7th day after grouping and scald+medium-dose group both on the 3rd and 7th day after grouping were significantly increased (P<0.05 or P<0.01), and the levels of TNF-α in ileal mucosa tissue of rabbits in scald+high-dose group on the 3rd and 7th day after grouping and in scald+medium-dose group on the 7th day after grouping were all significantly decreased (P<0.05 or P<0.01), and the level of IL-1β in ileal mucosa tissue of rabbits in scald+medium-dose group on the 7th day after grouping was significantly decreased (P<0.01), and the level of IL-10 in ileal mucosa tissue of rabbits in scald alone group on the 7th day after grouping was significantly decreased (P<0.01). Compared with that on the 3rd day after grouping, the levels of TNF-α and IL-1β in ileal mucosa tissue of rabbits in scald alone group and the levels of IL-10 in ileal mucosa tissue of rabbits in normal control group, scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 7th day after grouping were all significantly increased (P<0.05 or P<0.01); and the levels of TNF-α in ileal mucosa tissue of rabbits in scald+low-dose group, scald+medium-dose group, and scald+high-dose group on the 7th day after grouping were all significantly decreased (P<0.05), and the levels of IL-1β in ileal mucosa tissue of rabbits both in scald+medium-dose group and scald+high-dose group on the 7th day after grouping were significantly decreased (P<0.05 or P<0.01), and the levels of IL-10 in ileal mucosa tissue of rabbits in scald alone group on the 7th day after grouping was significantly decreased (P<0.01). On the 7th day after grouping, the high-quality sequences obtained from the microflora in ileum mucosa of rabbits in normal control group, scald alone group, and scald+medium-dose group were 96 023, 107 365, and 95 921, respectively. At the classification level of phylum, class, order, family, and genus of the microflora in ileum mucosa of rabbits in three groups were all Bacteroidetes and Firmicutes, Clostridium and Bacteroidetes, Clostridium and Bacteroidetes, Rumenobacteriaceae and Clostridium and Bacteroideaceae, Clostridium and Bacteroidetes and rumen bacteria mainly, while the percentage of microflora in each group was different. There were no significant differences in Ace, Chao1, Simpson, Shannon indices (P>0.05), and no obvious difference in β diversity of microflora in ileal mucosa tissue of rabbits among three groups. Conclusions: After severe scalding, the inflammatory response of rabbit ileal mucosa tissue is obvious and increased in a time-dependent manner. Modified Sijunzi Decoction can reduce inflammation with optimal therapeutic concentration of 1.0 g/mL. The technology of high-throughput sequencing can reflect the structural composition of the intestinal microflora accurately. The ileal microflora of the severe scald rabbit can be regulated by the administration of Modified Sijunzi Decoction.