Qiangchuan Hou

Intestinal Microbiota Distinguish Gout Patients from Healthy Humans

Current blood-based approach for gout diagnosis can be of low sensitivity and hysteretic. Here via a 68-member cohort of 33 healthy and 35 diseased individuals, we reported that the intestinal microbiota of gout patients are highly distinct from healthy individuals in both organismal and functional structures. In gout, Bacteroides caccae and Bacteroides xylanisolvens are enriched yet Faecalibacterium prausnitzii and Bifidobacterium pseudocatenulatum depleted. The established reference microbial gene catalogue for gout revealed disorder in purine degradation and butyric acid biosynthesis in gout patients. In an additional 15-member validation-group, a diagnosis model via 17 gout-associated bacteria reached 88.9% accuracy, higher than the blood-uric-acid based approach. Intestinal microbiota of gout are more similar to those of type-2 diabetes than to liver cirrhosis, whereas depletion of Faecalibacterium prausnitzii and reduced butyrate biosynthesis are shared in each of the metabolic syndromes. Thus the Microbial Index of Gout was proposed as a novel, sensitive and non-invasive strategy for diagnosing gout via fecal microbiota.

High-throughput sequencing for the detection of the bacterial and fungal diversity in Mongolian naturally fermented cow’s milk in Russia

BackgroundTraditional fermented dairy products are major components of the typical Mongolian diet since ancient times. However, almost all the previous studies on the microbial composition of traditional Mongolian fermented dairy products analyzed food samples from the Chinese Mongolian region and Mongolia but not the Russian Mongolian region. In this study, the bacterial and fungal community diversity of nineteen naturally fermented cow’s milk (NFCM) samples from local Mongolian families residing in Kalmykia and Chita of Russia was investigated with pyrosequencing.ResultsFirmicutes and Ascomycota were the predominant phyla respectively for bacteria and fungi. The abundance of the bacterial phylum Acidobacteria was considerably different between the samples from the two regions. At genus level, Lactobacillus and Pichia were the predominating bacterial and fungal genera, respectively, while six bacterial genera significantly differed between the Kalmykia (enrichment of Aeromonas, Bacillus, Clostridium, Streptococcus, Vogesella) and Chita (enrichment of Lactococcus) samples. The results of principal coordinate analysis (PCoA) based on the bacterial or fungal composition of the Kalmykia and Chita samples revealed a different microbiota structure between the samples collected in these two locations. The redundancy analysis (RDA) identified 60 bacterial and 21 fungal OTUs as the key variables responsible for such microbiota structural difference.ConclusionsOur results suggest that structural differences existed in the microbiota of NFCM between Kalmykia and Chita. The difference in geographic environment may be an important factor influencing the microbial diversity of NFCM made by the Mongolians in Russia.

Evaluation of bacterial contamination in raw milk, ultra-high temperature milk and infant formula using single molecule, real-time sequencing technology.

The Pacific Biosciences (Menlo Park, CA) single molecule, real-time sequencing technology (SMRT) was reported to have some advantages in analyzing the bacterial profile of environmental samples. In this study, the presence of bacterial contaminants in raw milk, UHT milk, and infant formula was determined by SMRT sequencing of the full length 16S rRNA gene. The bacterial profiles obtained at different taxonomic levels revealed clear differences in bacterial community structure across the 16 analyzed dairy samples. No indicative pathogenic bacteria were found in any of these tested samples. However, some of the detected bacterial species (e.g., Bacillus cereus, Enterococcus casseliflavus, and Enterococcus gallinarum) might potentially relate with product quality defects and bacterial antibiotic gene transfer. Although only a limited number of dairy samples were analyzed here, our data have demonstrated for the first time the feasibility of using the SMRT sequencing platform in detecting bacterial contamination. Our paper also provides interesting reference information for future development of new precautionary strategies for controlling the dairy safety in large-scale industrialized production lines.

Investigation of bacterial and fungal diversity in tarag using high-throughput sequencing

This is the first study on the bacterial and fungal community diversity in 17 tarag samples (naturally fermented dairy products) through a metagenomic approach involving high-throughput pyrosequencing. Our results revealed the presence of a total of 47 bacterial and 43 fungal genera in all tarag samples, in which Lactobacillus and Galactomyces were the predominant genera of bacteria and fungi, respectively. The number of some microbial genera, such as Lactococcus, Acetobacter, Saccharomyces, Trichosporon, and Kluyveromyces, among others, was found to vary between different samples. Altogether, our results showed that the microbial flora in different samples may be stratified by geographic region.

Assessment of the bacterial and fungal diversity in home-made yoghurts of Xinjiang, China by pyrosequencing.

BACKGROUND The aim of this study was to gain a deeper knowledge of the bacterial and fungal community diversity in local home-made yoghurts from Zhaosu and Tekesi counties, Xinjiang, China, where a relatively high proportion of the ethnic minority population resides. RESULTS The bacterial and fungal community diversity in 22 home-made yoghurt samples was analyzed by pyrosequencing. This approach revealed the presence of six bacterial and two fungal phyla, comprising 69 bacterial and 20 fungal genera respectively, among all samples. Firmicutes and Ascomycota were the dominant phyla and Lactobacillus and Saccharomyces the predominant genera of bacteria and fungi respectively. Based on redundancy analysis, the bacterial diversity in samples from the two counties differed mainly because of eight responsive operational taxonomic units of Lactobacillus. However, no obvious difference existed in the fungal population of samples from the two counties. CONCLUSION The results indicated that the microbial community of home-made yoghurts varied with their geographical origin and manufacturing process. The study showed that pyrosequencing could provide a better understanding of microbiological diversity as well as useful information for future studies on the selection of starter cultures for yoghurts.

High-throughput sequencing reveals microbial community diversity of Tibetan naturally fermented yak milk

Naturally fermented yak milk (NFYM) is a traditional Tibetan fermented product that contains a rich microbial community. This study used high-throughput pyrosequencing to investigate the bacterial and fungal community diversity of 16 NFYM samples from the villages Geda and Ningzhong in Tibet. Pyrosequencing produced a total of 112,173 high quality bacterial 16S rRNA gene sequences and 90,980 fungal high quality reads from the NFYM. Sequence analysis grouped the high quality gene sequences into eleven bacterial and five fungal phyla, of which Firmicutes and Ascomycota dominated. At the genus level, Lactobacillus and Saccharomyces were the dominating bacteria and fungi, respectively. Principal coordinate analysis (PCoA) based on weighted and unweighted UniFrac values revealed no significant differences in the structure of bacterial and fungal microbiota. However, multivariate analysis of variance (MANOVA) based on unweighted UniFrac distances showed significant differences (P < 0.05) between the bacterial and fungal populations of the two NFYM samples from different villages. After the redundancy analysis (RDA), 49 key responding operational taxonomic units (OTUs) could be identified. Among them, 7 OTUs (Acinetobacter, unidentified Bacteroidetes, Lactobacillus, unidentified Proteobacteria, Streptococcus, Pantoea, and unidentified Firmicutes) were enriched in the Geda village samples, while 42 OTUs (mainly Massilia, Propionibacterium, Lactococcus, Leuconostoc, and Enterococcus) were enriched in the Ningzhong village samples. The present work provided insights into the microbiome of Tibetan NFYM and showed that the microbiota composition may be stratified by their geographic regions.

Unique Features of Ethnic Mongolian Gut Microbiome revealed by metagenomic analysis

The human gut microbiota varies considerably among world populations due to a variety of factors including genetic background, diet, cultural habits and socioeconomic status. Here we characterized 110 healthy Mongolian adults gut microbiota by shotgun metagenomic sequencing and compared the intestinal microbiome among Mongolians, the Hans and European cohorts. The results showed that the taxonomic profile of intestinal microbiome among cohorts revealed the Actinobaceria and Bifidobacterium were the key microbes contributing to the differences among Mongolians, the Hans and Europeans at the phylum level and genus level, respectively. Metagenomic species analysis indicated that Faecalibacterium prausnitzii and Coprococcus comeswere enrich in Mongolian people which might contribute to gut health through anti-inflammatory properties and butyrate production, respectively. On the other hand, the enriched genus Collinsella, biomarker in symptomatic atherosclerosis patients, might be associated with the high morbidity of cardiovascular and cerebrovascular diseases in Mongolian adults. At the functional level, a unique microbial metabolic pathway profile was present in Mongolian’s gut which mainly distributed in amino acid metabolism, carbohydrate metabolism, energy metabolism, lipid metabolism, glycan biosynthesis and metabolism. We can attribute the specific signatures of Mongolian gut microbiome to their unique genotype, dietary habits and living environment.

A Perspective Study of Koumiss Microbiome by Metagenomics Analysis Based on Single-Cell Amplification Technique

Koumiss is a traditional fermented dairy product and a good source for isolating novel bacteria with biotechnology potential. In the present study, we applied the single-cell amplification technique in the metagenomics analysis of koumiss. This approach aimed at detecting the low-abundant bacteria in the koumiss. Briefly, each sample was first serially diluted until reaching the level of approximately 100 cells. Then, three diluted bacterial suspensions were randomly picked for further study. By analyzing 30 diluted koumiss suspensions, a total of 24 bacterial species were identified. In addition to the previously reported koumiss-associated species, such as Lactobacillus (L.) helveticus. Lactococcus lactis. L. buchneri, L. kefiranofaciens, and Acetobacter pasteurianus, we successfully detected three low-abundant taxa in the samples, namely L. otakiensis. Streptococcus macedonicus, and Ruminococcus torques. The functional koumiss metagenomes carried putative genes that relate to lactose metabolism and synthesis of typical flavor compounds. Our study would encourage the use of modern metagenomics to discover novel species of bacteria that could be useful in food industries.

Differential fecal microbiota are retained in broiler chicken lines divergently selected for fatness traits

Our study combined 16S rRNA-pyrosequencing and whole genome sequencing to analyze the fecal metagenomes of the divergently selected lean (LL) and fat (FL) line chickens. Significant structural differences existed in both the phylogenic and functional metagenomes between the two chicken lines. At phylum level, the FL group had significantly less Bacteroidetes. At genus level, fourteen genera of different relative abundance were identified, with some known short-chain fatty acid producers (including Subdoligranulum, Butyricicoccus, Eubacterium, Bacteroides, Blautia) and a potentially pathogenic genus (Enterococcus). Redundancy analysis identified 190 key responsive operational taxonomic units (OTUs) that accounted for the structural differences between the phylogenic metagenome of the two groups. Four Cluster of Orthologous Group (COG) categories (Amino acid transport and metabolism, E; Nucleotide transport and metabolism, F; Coenzyme transport and metabolism, H; and Lipid transport and metabolism, I) were overrepresented in LL samples. Fifteen differential metabolic pathways (Biosynthesis of amino acids, Pyruvate metabolism, Nitrotoluene degradation, Lipopolysaccharide biosynthesis, Peptidoglycan biosynthesis, Pantothenate and CoA biosynthesis, Glycosaminoglycan degradation, Thiamine metabolism, Phosphotransferase system, Two-component system, Bacterial secretion system, Flagellar assembly, Bacterial chemotaxis, Ribosome, Sulfur relay system) were identified. Our data highlighted interesting variations between the gut metagenomes of these two chicken lines.

The impact of oral consumption of Lactobacillus plantarum P-8 on faecal bacteria revealed by pyrosequencing

Lactobacillus plantarum P-8 (Lp-8) is a novel probiotic bacterium showing beneficial effects in human. The current study is a follow-up work of a previous clinical trial, and it aimed at understanding the mechanism of the observed desirable effects by evaluating the change in faecal bacterial structure at phylogenetic level. The genomic DNAs used for 16S rRNA polymerase chain reaction (PCR)-pyrosequencing were obtained from the previous clinical trial. Briefly, 33 recruited subjects were given a single daily oral dose of Lp-8 (6×10(10) cfu) for 4 weeks. Faecal genomic DNAs were extracted from samples collected before (day 0) and after (day 28) Lp-8 consumption from the subjects. Our results show that the consumption of Lp-8 did not result in major change in the faecal bacterial structure by principal coordinate analysis. However, multivariate analysis of variance and analysis of UniFrac metric distances at day 0 and day 28 revealed significant difference in the faecal bacterial structure upon Lp-8 treatment. Thus, the difference lied on the less abundant rather than the predominant lineages. At genus level, the relative abundance of 5 Firmicutes genera (Leuconostoc, Lactobacillus, Sporacetigenium, Blautia and Staphylococcus) significantly increased after Lp-8 consumption, whereas 3 Proteobacteria genera (Shigella, Escherichia and Enterobacter) decreased (P<0.05). 60 key responding lineages could be identified at operational taxonomic unit (OTU) level by redundancy analysis. Among them, the abundance of 18 OTUs showed significant difference (P<0.05), while 12 OTUs exhibited modest change (0.05