Characterisation of microbiota in saliva, bronchoalveolar lavage fluid, non-malignant, peritumoural and tumour tissue in non-small cell lung cancer patients: a cross-sectional clinical trial.

BACKGROUND: While well-characterised on its molecular base, non-small cell lung cancer (NSCLC) and its interaction with local microbiota remains scarcely explored. Moreover, current studies vary in source of lung microbiota, from bronchoalveolar lavage fluid (BAL) to tissue, introducing potentially differing results. Therefore, the objective of this study was to provide detailed characterisation of the oral and multi-source lung microbiota of direct interest in lung cancer research. Since lung tumours in lower lobes (LL) have been associated with decreased survival, characteristics of the microbiota in upper (UL) and lower tumour lobes have also been examined. METHODS: Using 16S rRNA gene sequencing technology, we analysed microbiota in saliva, BAL (obtained directly on excised lobe), non-malignant, peritumoural and tumour tissue from 18 NSCLC patients eligible for surgical treatment. Detailed taxonomy, diversity and core members were provided for each microbiota, with analysis of differential abundance on all taxonomical levels (zero-inflated binomial general linear model with Benjamini-Hochberg correction), between samples and lobe locations. RESULTS: Diversity and differential abundance analysis showed clear separation of oral and lung microbiota, but more importantly, of BAL and lung tissue microbiota. Phylum Proteobacteria dominated tissue samples, while Firmicutes was more abundant in BAL and saliva (with class Clostridia and Bacilli, respectively). However, all samples showed increased abundance of phylum Firmicutes in LL, with decrease in Proteobacteria. Also, clades Actinobacteria and Flavobacteriia showed inverse abundance between BAL and extratumoural tissues depending on the lobe location. While tumour microbiota seemed the least affected by location, peritumoural tissue showed the highest susceptibility with markedly increased similarity to BAL microbiota in UL. Differences between the three lung tissues were however very limited. CONCLUSIONS: Our results confirm that BAL harbours unique lung microbiota and emphasise the importance of the sample choice for lung microbiota analysis. Further, limited differences between the tissues indicate that different local tumour-related factors, such as tumour type, stage or associated immunity, might be the ones responsible for microbiota-shaping effect. Finally, the "shift" towards Firmicutes in LL might be a sign of increased pathogenicity, as suggested in similar malignancies, and connected to worse prognosis of the LL tumours. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT03068663. Registered February 27, 2017.

The human intestinal microbiota of constipated-predominant irritable bowel syndrome patients exhibits anti-inflammatory properties.

The intestinal microbiota of patients with constipated-predominant irritable bowel syndrome (C-IBS) displays chronic dysbiosis. Our aim was to determine whether this microbial imbalance instigates perturbation of the host intestinal mucosal immune response, using a model of human microbiota-associated rats (HMAR) and dextran sulfate sodium (DSS)-induced experimental colitis. The analysis of the microbiota composition revealed a decrease of the relative abundance of Bacteroides, Roseburia-Eubacterium rectale and Bifidobacterium and an increase of Enterobacteriaceae, Desulfovibrio sp., and mainly Akkermansia muciniphila in C-IBS patients compared to healthy individuals. The bacterial diversity of the gut microbiota of healthy individuals or C-IBS patients was maintained in corresponding HMAR. Animals harboring a C-IBS microbiota had reduced DSS colitis with a decreased expression of pro-inflammatory cytokines from innate, Th1, and Th17 responses. The pre-treatment of conventional C57BL/6 mice or HMAR with A. muciniphila, but not with Escherichia coli, prior exposure to DSS also resulted in a reduction of colitis severity, highlighting that the anti-inflammatory effect of the gut microbiota of C-IBS patients is mediated, in part, by A. muciniphila. This work highlights a novel aspect of the crosstalk between the gut microbiota of C-IBS patients and host intestinal homeostasis.

Ruminococcus champanellensis sp. nov., a cellulose-degrading bacterium from human gut microbiota.

A strictly anaerobic, cellulolytic strain, designated 18P13(T), was isolated from a human faecal sample. Cells were Gram-positive non-motile cocci. Strain 18P13(T) was able to degrade microcrystalline cellulose but the utilization of soluble sugars was restricted to cellobiose. Acetate and succinate were the major end products of cellulose and cellobiose fermentation. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Ruminococcus of the family Ruminococcaceae. The closest phylogenetic relative was the ruminal cellulolytic strain Ruminococcus flavefaciens ATCC 19208(T) (<95% 16S rRNA gene sequence similarity). The DNA G+C content of strain 18P13(T) was 53.05±0.7 mol%. On the basis of phylogenetic analysis, and morphological and physiological data, strain 18P13(T) can be differentiated from other members of the genus Ruminococcus with validly published names. The name Ruminococcus champanellensis sp. nov. is proposed, with 18P13(T) (=DSM 18848(T)=JCM 17042(T)) as the type strain.

The cellulose-degrading microbial community of the human gut varies according to the presence or absence of methanogens.

Cellulose-degrading microorganisms involved in the breakdown of plant cell wall material in the human gut remain rather unexplored despite their role in intestinal fermentation. Microcrystalline cellulose-degrading bacteria were previously identified in faeces of methane-excreting individuals, whereas these microorganisms were undetectable in faecal samples from non-methane excretors. This suggested that the structure and activity of the cellulose-degrading community differ in methane- and non-methane-excreting individuals. The purpose of this study was to characterize in depth this cellulose-degrading community in individuals of both CH(4) statuses using both culture-dependent and molecular methods. A new real-time PCR analysis was developed to enumerate microcrystalline cellulose-degrading ruminococci and used to confirm the predominance of these hydrolytic ruminococci in methane excretors. Culture-dependent methods using cell wall spinach (CWS) residue revealed the presence of CWS-degrading microorganisms in all individuals. Characterization of CWS-degrading isolates further showed that the main cellulose-degrading bacteria belong essentially to Bacteroidetes in non-methane-excreting subjects, while they are predominantly represented by Firmicutes in methane-excreting individuals. This taxonomic diversity was associated with functional diversity: the ability to degrade different types of cellulose and to produce H(2) from fermentation differed depending on the species. The structure of the cellulolytic community was shown to vary depending on the presence of methanogens in the human gut.

Bacteroides xylanisolvens sp. nov., a xylan-degrading bacterium isolated from human faeces.

During the course of a study on the xylan-degrading community from the human gut, six xylanolytic, Gram-negative, anaerobic rods were isolated from faecal samples. 16S rRNA gene sequence analysis showed that the isolates were closely related to each other (> or =99 % sequence similarity) and that they belonged to the genus Bacteroides. On the basis of 16S rRNA gene sequence similarity, representative strain XB1AT was most closely related to the type strains of Bacteroides ovatus (97.5 %), B. finegoldii (96.5 %) and B. thetaiotaomicron (95.5 %). DNA-DNA hybridization results revealed that strain XB1AT was distinct from its closest relative, B. ovatus. The DNA G+C content of strain XB1AT (42.8 mol%) and major fatty acid composition (anteiso-C15 : 0, 33.8 %) further supported its affiliation to the genus Bacteroides. The novel isolates degraded different types of xylan, and were also able to grow on a variety of carbohydrates. Unlike most other Bacteroides species isolated from the human gut, these isolates were not able to degrade starch. Other biochemical tests further demonstrated that strain XB1AT could be differentiated from the closest related Bacteroides species. Xylan and sugars were converted by strain XB1AT mainly into acetate, propionate and succinate. Based on physiological, phenotypic and phylogenetic data, the six novel strains are considered to represent a novel species of the genus Bacteroides, for which the name Bacteroides xylanisolvens sp. nov. is proposed. The type strain is XB1AT (=DSM 18836T =CCUG 53782T).