Ultra-high-depth macrogenomic sequencing revealed differences in microbial composition and function between high temperature and medium-high temperature Daqu.

Complex microorganisms in Daqu of different temperatures play a vital role in the taste, flavor and quality of Baijiu during fermentation. However, understanding the functional diversity of the whole microbial community between the Daqus of two different temperatures (high temperature Daqu, HD and medium-high temperature Daqu, MD) remains a major challenge. Here, a systematic study of the microbial diversity, functions as well as physiological and biochemical indexes of Daqu are described. The results revealed that the Daqu exhibited unique characteristics. In particular, the diversity of microorganisms in HD and MD was high, with 44 species including 14 novel species (Sphingomonas sp. is the main novel species) detected in all samples. Their profiles of carbohydrate-active enzymes and specific functional components supported the fact that these species were involved in flavor formation. The Daqu microbiome consisted of a high proportion of phage, providing evidence of phage infection/genome integration and horizontal gene transfer from phage to bacteria. Such processes would also regulate Daqu microbiomes and thus flavor quality. These results enrich current knowledge of Daqu and can be used to promote the development of Baijiu fermentation technology.

Analysis of bacterial diversity and genetic evolution of Lacticaseibacillus paracasei isolates in fermentation pit mud.

AIMS: Since little is known about the genetic diversity of lactic acid bacteria (LAB) isolates from the fermentation pit mud (FPM), we sought to evaluate the bacterial structure, identify the LAB isolates and investigate the genotype and genetic diversity of the LAB isolates. METHODS AND RESULTS: Using high-throughput MiSeq sequencing, we identified seven dominant bacterial genera in FPM. Lactobacillus had the highest abundance. We isolated 55 LAB strains. These isolates were all identified as Lacticaseibacillus paracasei. Using an extant multilocus sequence typing (MLST) scheme, isolates were assigned to 18 sequence types (STs) and three clonal complexes. ST1, the largest group, mainly comprised FPM isolates. Niche-specific ST2 to ST18 only contained FPM isolates. Isolates could be divided into four lineages, with most assigned to Lineage 1. Only one FPM isolate was classified as L. paracasei subsp. paracasei. Other isolates could not be classified at the subspecies level using the seven MLST loci. CONCLUSIONS: Lactobacilli account for a high proportion of bacteria in pit mud. Based on the traditional culture method, L. paracasei was the dominant species, and these isolates exhibit a high ethanol tolerance, high intraspecific diversity and specific genetic profiles. SIGNIFICANCE AND IMPACT OF THE STUDY: The study described the characterization of FPM bacterial diversity, giving an insight into the genetic diversity of L. paracasei strains present in FPM.

PacBio sequencing revealed variation in the microbiota diversity, species richness and composition between milk collected from healthy and mastitis cows.

Mastitis is the economically most important disease of dairy cows. This study used PacBio single-molecule real-time sequencing technology to sequence the full-length 16S rRNAs from 27 milk samples (18 from mastitis and nine from healthy cows; the cows were at different stages of lactation). We observed that healthy or late stage milk microbiota had significantly higher microbial diversity and richness. The community composition of the microbiota of different groups also varied greatly. The healthy cow milk microbiota was predominantly comprised of Lactococcus lactis, Acinetobacter johnsonii, and Bacteroides dorei, while the milk from mastitis cows was predominantly comprised of Bacillus cereus. The prevalence of L. lactis and B. cereus in the milk samples was confirmed by digital droplets PCR. Differences in the milk microbiota diversity and composition could suggest an important role for some these microbes in protecting the host from mastitis while others associated with mastitis. The results of our research serve as useful references for designing strategies to prevent and treat mastitis.

Bacterial Diversity in Pickled Cowpea (Vigna unguiculata [Linn.] Walp) as Determined by Illumina MiSeq Sequencing and Culture-Dependent Methods.

Pickled cowpea (Vigna unguiculata [Linn.] Walp) is a popular fermented vegetable in China that is made by spontaneous fermentation. Prior to this study, little was known about its microbial community. Eighteen pickled cowpea samples were collected in Enshi City, China, in 2018. The bacterial diversity within these samples was evaluated using a combination of high-throughput sequencing (Illumina MiSeq platform) targeting the V3-V4 region of the 16S rRNA gene sequence and culture-dependent methods. A total of 456,318 high-quality 16S rRNA gene sequence reads were obtained, and these reads were clustered into 19,712 OTUs with 97.0% similarity. The core bacterial phyla were Actinobacteria, Bacteroidetes, Proteobacteria, and Firmicutes; the core bacterial genera were Levilactobacillus, Lactiplantibacillus, Companilactobacillus, Pediococcus, Lactobacillus, Weissella, and Pseudomonas. Using the spread-plating method, 39 lactic acid bacteria (LAB) strains were isolated and identified based on the nearly complete 16S rRNA gene sequence. Of these, 37 were identified as Lactiplantibacillus plantarum group, while the other two were classified as Limosilactobacillus fermentum and Lacticaseibacillus rhamnosus. These results indicate a high relative abundance of LAB in traditional pickled cowpea, especially Lactobacillaceae species, which likely contribute to fermentation. This study would provide information on the LAB population of Pickled cowpea and indicated that the Pickled cowpea could be a good source for isolation of lactic acid bacteria.

Profiling of koumiss microbiota and organic acids and their effects on koumiss taste.

BACKGROUND: Koumiss is a naturally fermented mare's milk. Over recent decades, numerous studies have revealed the diversity of lactic acid bacteria in koumiss. However, there is limited information available regarding its secondary major component yeast profile. RESULTS: A total of 119 bacterial and 36 yeast species were identified among the 14 koumiss samples. The dominant bacterial species in koumiss were Lactobacillus helveticus, Lactobacillus kefiranofaciens, Lactococcus lactis, Lactococcus raffinolactis, and Citrobacter freundii. The main yeast species were Dekkera anomala, Kazachstania unispora, Meyerozyma caribbica, Pichia sp.BZ159, Kluyveromyces marxianus, and uncultured Guehomyces. The bacterial and yeast Shannon diversity of the Xilinhaote-urban group were higher than those of the Xilingol-rural group. The most dominant organic acids were lactic, acetic, tartaric, and malic acids. Lactic acid bacteria species were mostly responsible for the accumulation of those organic acids, although Kazachstania unispora, Dekkera anomala, and Meyerozyma caribbica may also have contributed. Redundancy analysis suggested that both bacteria and yeast respond to koumiss flavor, such as Lactobacillus helveticus and Dekkera anomala are associated with sourness, astringency, bitterness, and aftertaste, whereas Lactococcus lactis and Kazachstania unispora are associated with umami. CONCLUSIONS: Our results suggest that differences were observed in koumiss microbiota of Xilinhaote-urban and Xilingol-rural samples. The biodiversity of the former was higher than the latter group. Positive or negative correlations between bacteria and yeast species and taste also were found.

Lactobacillus enshiensis sp. nov., a novel arsenic-resistant bacterium.

A lactic acid bacterial strain, HBUAS57009T, isolated from traditionally fermented food (Zha-Chili) in China, was characterized to clarify its taxonomic status using a polyphasic approach. Strain HBUAS57009T was phylogenetically closely related to Lactobacillus koreensis DCY50T, Lactobacillus fujinensis 218-6T, Lactobacillus mulengensis 112-3T, Lactobacillus cerevisiae TUM BP 140423000-2250T, Lactobacillus tongjiangensis 218-10T and Lactobacillus yonginensis THK-V8T with sequence similarities of 98.6-99.3 %. The genome-to-genome distance and average nucleotide identity values between the genomes of strain HBUAS57009T and type strains of closely related Lactobacillus species were less than 32.0 and 86.0 %, respectively; this is below the threshold for species boundaries. The major cellular fatty acids (>10 %) were C16 : 0, C18 : 1 ω9c and iso-C19 : 0. The G+C content of the genomic DNA of strain HBUAS57009T was 47.8 mol%. Examination of the functional categories of the genome revealed that strain HBUAS57009T could perform both homolactic and heterolactic fermentation processes to produce lactic acid via complete glycolysis and the pentose phosphate pathway. The putative biosynthesis pathway of butane-2,3-diol and acetoin, two important flavour compounds in the food industry, were identified using kegg mapper analysis. Based on its genotypic and phenotypic features, strain HBUAS57009T (=GDMCC 1.1664T=KACC 21424T) is designated as the type strain of a novel species, for which the name Lactobacillus enshiensis sp. nov. is proposed.

Lactobacillus casei protects dextran sodium sulfate- or rapamycin-induced colonic inflammation in the mouse.

PURPOSE: Human colon inflammation is associated with changes in the diverse and abundant microorganisms in the gut. As important beneficial microbes, Lactobacillus contributes to the immune responses and intestinal integrity that may alleviate experimental colitis. However, the mechanisms underlying probiotic benefits have not been fully elucidated. METHODS: Dextran sodium sulfate or rapamycin-challenged mice were used as model for colon inflammation evaluation. Histological scores of the colon, levels of colonic myeloperoxidase, serum tumor necrosis factor-α and interleukin-6 were assessed as inflammatory markers and the gut microbiota profiles of each mouse were studied. RESULTS: We found that Lactobacillus casei Zhang (LCZ) can prevent experimental colitis and rapamycin-induced inflammation in intestinal mucosa by improving histological scores, decreasing host inflammatory cytokines, modulating gut-dominated bacteria, enhancing cystic fibrosis transmembrane conductance regulator (CFTR) expression and downregulating the expression of p-STAT3 (phosphorylated signal transducer and activator of transcription 3) or Akt/NF-κB (AKT serine/threonine kinase and nuclear factor kappa B). CONCLUSION: Our results suggest that LCZ may provide effective prevention against colitis.

PacBio sequencing reveals bacterial community diversity in cheeses collected from different regions.

Cheese is a fermented dairy product that is popular for its unique flavor and nutritional value. Recent studies have shown that microorganisms in cheese play an important role in the fermentation process and determine the quality of the cheese. We collected 12 cheese samples from different regions and studied the composition of their bacterial communities using PacBio small-molecule real-time sequencing (Pacific Biosciences, Menlo Park, CA). Our data revealed 144 bacterial genera (including Lactobacillus, Streptococcus, Lactococcus, and Staphylococcus) and 217 bacterial species (including Lactococcus lactis, Streptococcus thermophilus, Staphylococcus equorum, and Streptococcus uberis). We investigated the flavor quality of the cheese samples using an electronic nose system and we found differences in flavor-quality indices among samples from different regions. We found a clustering tendency based on flavor quality using principal component analysis. We found correlations between lactic acid bacteria and the flavor quality of the cheese samples. Biodegradation and metabolism of xenobiotics, and lipid-metabolism-related pathways, were predicted to contribute to differences in cheese flavor using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt). This preliminary study explored the bacterial communities in cheeses collected from different regions and their potential genome functions from the perspective of flavor quality.

Comparative genomic analysis revealed great plasticity and environmental adaptation of the genomes of Enterococcus faecium.

BACKGROUND: As an important nosocomial pathogen, Enterococcus faecium has received increasing attention in recent years. However, a large number of studies have focused on the hospital-associated isolates and ignored isolates originated from the natural environments. RESULTS: In this study, comparative genomic analysis was conducted on 161 isolates originated from human, animal, and naturally fermented dairy products. The results showed that the environment played an important role in shaping the genomes of Enterococcus faecium. The isolates from human had the largest average genome size, while the isolates from dairy products had the smallest average genome size and fewest antibiotic resistance genes. A phylogenetic tree was reconstructed based on the genomes of these isolates, which revealed new insights into the phylogenetic relationships among the dairy isolates and those from hospitals, communities, and animals. Furthermore, 202 environment-specific genes were identified, including 136 dairy-specific, 31 human blood-specific, and 35 human gastrointestinal-specific genes. Interestingly, five dairy-specific genes (namely lacF, lacA/B, lacD, lacG, and lacC) that constituted an integrated lactose metabolism pathway existed in almost all dairy isolates. The pathway conservation demonstrated an active role of the environment in shaping the genomes of Enterococcus faecium. CONCLUSIONS: This study shows that the Enterococcus faecium species has great genomic plasticity and high versatility to occupy broad ecological roles, dwelling as non-harmful dairy and animal gut commensals as well as significant nosocomial pathogens that disseminate antibiotic resistance genes.

Lactobacillus zhachilii sp. nov., a lactic acid bacterium isolated from Zha-Chili.

Strain HBUAS52074T is a Gram-positive staining, aerobic bacterium that was isolated from Zha-Chili, a traditional fermented food made in China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain HBUAS52074T is a member of the genus Lactobacillus and closely related to Lactobacillus nantensis DSM 16982T (98.9 %), Lactobacillus heilongjiangensis DSM 28069T (98.8 %), Lactobacillus formosensis NBRC 1095009T (98.6 %), Lactobacillus futsaii JCM 17355T (98.5 %), Lactobacillus farciminis KCTC 3681T (98.5 %), Lactobacillus musae NBRC 112868T (98.5 %) and Lactobacillus crustorum LMG 23699T (98.4 %). The DNA G+C content is 36.3 mol%. The major cellular fatty acids are C16 : 0 (28.2 %), C18 : 1ω9c (30.5 %) and summed feature 7 (C19 : 1ω6c, and/or C19 : 1ω7c; 14.9 %). Average nucleotide identity and DNA-DNA hybridization (GGDC) values based on genomic comparisons between HBUAS52074T and related type species showed that the bacterium was significantly different from its closest relatives. Using polyphasic taxonomic analysis, we have shown that strain HBUAS52074T is a new species in the genus Lactobacillus, for which we propose the name Lactobacilluszhachilii sp. nov. The type strain is HBUAS52074T (=GDMCC 1.1417T=KCTC 21106T).

Vibrio zhugei sp. nov., a moderately halophilic bacterium isolated from pickling sauce.

Strain HBUAS61001T was isolated from the pickling sauce used to make a traditional fermented food product, datoucai, in China. The strain belonged to the genus Vibrio, but was placed in a clade separate from any known Vibrio species based on the 16S rRNA gene and MLSA results. The genome consisted of two chromosomes: chromosome I was 2 901 449 bp long with a G+C content of 45.4 mol%; and chromosome II was 1 107 930 bp long with a G+C content of 45.5 mol%. The most abundant fatty acids were C16 : 0 (28.1 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c, 29.4 %) and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c, 10.1 %). The isoprenoid quinones detected were Q7 and Q8. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. Strain HBUAS61001T could grow in the presence of up to 17 % NaCl. The calculated average nucleotide identity and in silico DNA-DNA hybridization (GGDC) values of the strain against the closest related type strains were all lower than 95 and 70 %, respectively. Putative genes in the genome associated with survival under high salinity stress were identified. Based on whole genome sequence analysis and phenotypic characteristics, strain HBUAS61001T is a new species in the genus Vibrio, and the name Vibrio zhugei (=GDMCC 1.1416T=KCTC 62784T) is proposed.

Halobasidium xiangyangense gen. nov., sp. nov., a new xylose-utilizing yeast in the family Cystobasidiaceae, isolated from the pickling sauce used to make Datoucai, a high-salt fermented food.

In this study, we describe a new genus and species of yeast with high-salt tolerance. The strain was isolated from the pickling sauce used to make Datoucai, a traditional fermented food made from Brassica juncea in Xiangyang, China. Phylogenetic analysis of sequences from the D1/D2 region of the LSU rRNA gene and from the ITS region demonstrated that the strain, reference HBUAS51001T, was most closely related to members of the genera Occultifur and Cystobasidium. However, the greatest similarities between the D1/D2 and ITS nucleotide sequences of strain HBUAS51001T and the most closely related type strains from Occultifur and Cystobasidium were only 91 and 92 %, respectively. This suggests that strain HBUAS51001T does not belong to any currently described species. Strain HBUAS51001T grew readily on media in which xylose was the sole carbon source. The major ubiquinone was Q9. The genome of strain HBUAS51001T was 42.42 Mb with a G+C content of 53.93 mol%. Three candidate genes associated with xylose metabolism were identified. On the basis of genotypic and phenotypic data, strain HBUAS51001T can be considered as both a new species and a new genus, for which the name Halobasidium xiangyangense gen. nov., sp. nov. is proposed. The type strain is HBUAS51001T (=KCTC27810T=GDMCC 2.231T=CCTCC AY 2018002T).

Investigating the bacterial microbiota of traditional fermented dairy products using propidium monoazide with single-molecule real-time sequencing.

Traditional fermented dairy foods have been the major components of the Mongolian diet for millennia. In this study, we used propidium monoazide (PMA; binds to DNA of nonviable cells so that only viable cells are enumerated) and single-molecule real-time sequencing (SMRT) technology to investigate the total and viable bacterial compositions of 19 traditional fermented dairy foods, including koumiss from Inner Mongolia (KIM), koumiss from Mongolia (KM), and fermented cow milk from Mongolia (CM); sample groups treated with PMA were designated PKIM, PKM, and PCM. Full-length 16S rRNA sequencing identified 195 bacterial species in 121 genera and 13 phyla in PMA-treated and untreated samples. The PMA-treated and untreated samples differed significantly in their bacterial community composition and α-diversity values. The predominant species in KM, KIM, and CM were Lactobacillus helveticus, Streptococcus parauberis, and Lactobacillus delbrueckii, whereas the predominant species in PKM, PKIM, and PCM were Enterobacter xiangfangensis, Lactobacillus helveticus, and E. xiangfangensis, respectively. Weighted and unweighted principal coordinate analyses showed a clear clustering pattern with good separation and only minor overlapping. In addition, a pure culture method was performed to obtain lactic acid bacteria resources in dairy samples according to the results of SMRT sequencing. A total of 102 LAB strains were identified and Lb. helveticus (68.63%) was the most abundant, in agreement with SMRT sequencing results. Our results revealed that the bacterial communities of traditional dairy foods are complex and vary by type of fermented dairy product. The PMA treatment induced significant changes in bacterial community structure.

Comparative genomic analysis of Enterococcus faecalis: insights into their environmental adaptations.

BACKGROUND: Enterococcus faecalis is widely studied as a common gut commensal and a nosocomial pathogen. In fact, Enterococcus faecalis is ubiquitous in nature, and it has been isolated from various niches, including the gastrointestinal tract, faeces, blood, urine, water, and fermented foods (such as dairy products). In order to elucidate the role of habitat in shaping the genome of Enterococcus faecalis, we performed a comparative genomic analysis of 78 strains of various origins. RESULTS: Although no correlation was found between the strain isolation habitat and the phylogeny of Enterococcus faecalis from our whole genome-based phylogenetic analysis, our results revealed some environment-associated features in the analysed Enterococcus faecalis genomes. Significant differences were found in the genome size and the number of predicted open reading frames (ORFs) between strains originated from different environments. In general, strains from water sources had the smallest genome size and the least number of predicted ORFs. We also identified 293 environment-specific genes, some of which might link to the adaptive strategies for survival in particular environments. In addition, the number of antibiotic resistance genes was significantly different between strains isolated from dairy products, water, and blood. Strains isolated from blood had the largest number of antibiotic resistance genes. CONCLUSION: These findings improve our understanding of the role of habitat in shaping the genomes of Enterococcus faecalis.

Analysis of the Gut Microbial Diversity of Dairy Cows During Peak Lactation by PacBio Single-Molecule Real-Time (SMRT) Sequencing.

BACKGROUND: The gut microbes of dairy cows are strongly associated with their health, but the relationship between milk production and the intestinal microbiota has seldom been studied. Thus, we explored the diversity of the intestinal microbiota during peak lactation of dairy cows. METHODS: The intestinal microbiota of nine dairy cows at peak lactation was evaluated using the Pacific Biosciences single-molecule real-time (PacBio SMRT) sequencing approach. RESULTS: A total of 32,670 high-quality 16S rRNA gene sequences were obtained, belonging to 12 phyla, 59 families, 107 genera, and 162 species. Firmicutes (83%) were the dominant phylum, while Bacteroides (6.16%) was the dominant genus. All samples showed a high microbial diversity, with numerous genera of short chain fatty acid (SCFA)-producers. The proportion of SCFA producers was relatively high in relation to the identified core intestinal microbiota. Moreover, the predicted functional metagenome was heavily involved in energy metabolism. CONCLUSIONS: This study provided novel insights into the link between the dairy cow gut microbiota and milk production.

Bacterial diversity and community structure in Chongqing radish paocai brines revealed using PacBio single-molecule real-time sequencing technology.

BACKGROUND: Traditional Chongqing radish paocai fermented with aged brine is considered to have the most intense flavor and authentic taste. Eight 'Yanzhi' (red, RRPB group) and 'Chunbulao' (white, WRPB) radish paocai brine samples were collected from Chongqing peasant households, and the diversity and community structures of bacteria present in these brines were determined using PacBio single-molecule real-time sequencing of their full-length 16S rRNA genes. RESULTS: In total, 30 phyla, 218 genera, and 306 species were identified from the RRPB group, with 20 phyla, 261 genera, and 420 species present in the WRPB group. Obvious differences in bacterial profiles between the RRPB and WRPB groups were found, with the bacterial diversity of the WRPB group shown to be greater than that of the RRPB group. This study revealed several characteristics of the bacteria composition, including the predominance of heterofermentative lactic acid bacteria, the species diversity of genus Pseudomonas, and the presence of three opportunistic pathogenic species. CONCLUSION: This study provides detailed information on the bacterial diversity and community structure of Chongqing radish paocai brine samples, and suggests it may be necessary to analyze paocai brine for potential sources of bacterial contamination and take appropriate measures to exclude any pathogenic species. © 2018 Society of Chemical Industry.

Bacterial microbiota of Kazakhstan cheese revealed by single molecule real time (SMRT) sequencing and its comparison with Belgian, Kalmykian and Italian artisanal cheeses.

BACKGROUND: In Kazakhstan, traditional artisanal cheeses have a long history and are widely consumed. The unique characteristics of local artisanal cheeses are almost completely preserved. However, their microbial communities have rarely been reported. The current study firstly generated the Single Molecule, Real-Time (SMRT) sequencing bacterial diversity profiles of 6 traditional artisanal cheese samples of Kazakhstan origin, followed by comparatively analyzed the microbiota composition between the current dataset and those from cheeses originated from Belgium, Russian Republic of Kalmykia (Kalmykia) and Italy. RESULTS: Across the Kazakhstan cheese samples, a total of 238 bacterial species belonging to 14 phyla and 140 genera were identified. Lactococcus lactis (28.93%), Lactobacillus helveticus (26.43%), Streptococcus thermophilus (12.18%) and Lactobacillus delbrueckii (12.15%) were the dominant bacterial species for these samples. To further evaluate the cheese bacterial diversity of Kazakhstan cheeses in comparison with those from other geographic origins, 16S rRNA datasets of 36 artisanal cheeses from Belgium, Russian Republic of Kalmykia (Kalmykia) and Italy were retrieved from public databases. The cheese bacterial microbiota communities were largely different across sample origins. By principal coordinate analysis (PCoA) and multivariate analysis of variance (MANOVA), the structure of the Kazakhstan artisanal cheese samples was found to be different from those of the other geographic origins. Furthermore, the redundancy analysis (RDA) identified 16 bacterial OTUs as the key variables responsible for such microbiota structural difference. CONCLUSION: Our results together suggest that the diversity of bacterial communities in different groups is stratified by geographic region. This study does not only provide novel information on the bacterial microbiota of traditional artisanal cheese of Kazakhstan at species level, but also interesting insights into the bacterial diversity of artisanal cheeses of various geographical origins.

Influence of Lactobacillus plantarum on yogurt fermentation properties and subsequent changes during postfermentation storage.

This study aimed to evaluate the influence of 9 Lactobacillusplantarum with broad-spectrum antibacterial activity on fermented milk, including changes to the fermentation characteristics (pH, titration acidity, and viable counts), texture profile, relative content of volatile compounds, and sensory evaluation during 28-d storage at 4°C. First, L. plantarum IMAU80106, IMAU10216, and IMAU70095 were selected as candidates for further study because of their excellent coagulation and proteolytic activities. Subsequently, these L. plantarum strainswere supplemented to fermented milk produced by commercial yogurt starters (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus) and a panel of parameters reflecting product quality was subsequently monitored during 28 d of postfermentation storage. The pH value and titration acidity of the fermented milk mildly fluctuated, whereas the L. plantarum viable counts remained stable along the storage period. Fourteen key volatile compounds were detected in the fermented milk by gas chromatography-mass spectrometry, and some flavor compounds were uniquely present in the L. plantarum-supplemented fermented milk (including 2,3-pentanedione, acetaldehyde, and acetate). No significant difference was shown in the sensory evaluation scores between samples with or without L. plantarum supplementation, but a gradual decrease was observed over storage in all samples. However, when L. plantarum was added, apparent shifts were observed in the overall quality of the fermented milk based on principal component analysis and multivariate ANOVA, particularly in the texture (adhesiveness) and volatile flavor compound profiles (acetaldehyde). Compared with L. plantarum IMAU80106 and IMAU10216, both the texture and volatile flavor profiles of IMAU70095 were closest to those of the control without adding the adjunct bacteria, suggesting that IMAU70095 might be the most suitable strain for further application in functional dairy product development. The current work has explored the potential of applying L. plantarum in fermented milk by performing thorough physical and chemical characterization. Our work is of intense interest to the dairy industry.

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

BACKGROUND: Traditional 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. RESULTS: Firmicutes 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. CONCLUSIONS: Our 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.