Microbial communities, functional, and flavor differences among three different-colored high-temperature Daqu: A comprehensive metagenomic, physicochemical, and electronic sensory analysis.

High-temperature Daqu (HTD) is the starter for producing sauce-flavor Baijiu, with different-colored Daqu (white, yellow, and black) reflecting variations in fermentation chamber conditions, chemical reactions, and associated microbiota. Understanding the relationship between Daqu characteristics and flavor/taste is challenging yet vital for improving Baijiu fermentation. This study utilized metagenomic sequencing, physicochemical analysis, and electronic sensory evaluation to compare three different-colored HTD and their roles in fermentation. Fungi and bacteria dominated the HTD-associated microbiota, with fungi increasing as the fermentation temperature rose. The major fungal genera were Aspergillus (40.17%) and Kroppenstedtia (21.16%), with Aspergillus chevalieri (25.65%) and Kroppenstedtia eburnean (21.07%) as prevalent species. Microbial communities, functionality, and physicochemical properties, particularly taste and flavor, were color-specific in HTD. Interestingly, the microbial communities in different-colored HTDs demonstrated robust functional complementarity. White Daqu exhibited non-significantly higher α-diversity compared to the other two Daqu. It played a crucial role in breaking down substrates such as starch, proteins, hyaluronic acid, and glucan, contributing to flavor precursor synthesis. Yellow Daqu, which experienced intermediate temperature and humidity, demonstrated good esterification capacity and a milder taste profile. Black Daqu efficiently broke down raw materials, especially complex polysaccharides, but had inferior flavor and taste. Notably, large within-group variations in physicochemical quality and microbial composition were observed, highlighting limitations in color-based HTD quality assessment. Water content in HTD was associated with Daqu flavor, implicating its crucial role. This study revealed the complementary roles of the three HTD types in sauce-flavor Baijiu fermentation, providing valuable insights for product enhancement.

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.

Effect of synbiotic supplementation on immune parameters and gut microbiota in healthy adults: a double-blind randomized controlled trial.

Synbiotics are increasingly used by the general population to boost immunity. However, there is limited evidence concerning the immunomodulatory effects of synbiotics in healthy individuals. Therefore, we conducted a double-blind, randomized, placebo-controlled study in 106 healthy adults. Participants were randomly assigned to receive either synbiotics (containing Bifidobacterium lactis HN019 1.5 × 108 CFU/d, Lactobacillus rhamnosus HN001 7.5 × 107 CFU/d, and fructooligosaccharide 500 mg/d) or placebo for 8 weeks. Immune parameters and gut microbiota composition were measured at baseline, mid, and end of the study. Compared to the placebo group, participants receiving synbiotic supplementation exhibited greater reductions in plasma C-reactive protein (P = 0.088) and interferon-gamma (P = 0.008), along with larger increases in plasma interleukin (IL)-10 (P = 0.008) and stool secretory IgA (sIgA) (P = 0.014). Additionally, synbiotic supplementation led to an enrichment of beneficial bacteria (Clostridium_sensu_stricto_1, Lactobacillus, Bifidobacterium, and Collinsella) and several functional pathways related to amino acids and short-chain fatty acids biosynthesis, whereas reduced potential pro-inflammatory Parabacteroides compared to baseline. Importantly, alternations in anti-inflammatory markers (IL-10 and sIgA) were significantly correlated with microbial variations triggered by synbiotic supplementation. Stratification of participants into two enterotypes based on pre-treatment Prevotella-to-Bacteroides (P/B) ratio revealed a more favorable effect of synbiotic supplements in individuals with a higher P/B ratio. In conclusion, this study suggested the beneficial effects of synbiotic supplementation on immune parameters, which were correlated with synbiotics-induced microbial changes and modified by microbial enterotypes. These findings provided direct evidence supporting the personalized supplementation of synbiotics for immunomodulation.

Depth-depended quality comparison of light-flavor fermented grains from two fermentation rounds.

To improve the utilization of raw materials, the fermentation of light-flavor Baijiu (LFB) will usually take two rounds of fermentation to the same batch of raw materials. The microorganisms involved in the fermented grains (FG) from these two fermentation rounds are the key to the quality formation of light-flavor style. Using Illumina MiSeq high-throughput sequencing and electronic-senses, it was found that the microbiota and flavor of FG for light-flavor Baijiu were mainly affected by fermentation rounds rather than depths, while the physicochemical factors were just the opposite. Starch was the most influential physiochemical factor on the microbiota. FG from the first fermentation round (FFRFG) had a higher fungal diversity, while FG from the second fermentation round (SFRFG) had a higher microbial richness and bacterial diversity. Monascus and the function of metabolism were significantly enriched in FFRFG; while Nocardia, Saccharomyces, Aspergillus and the functions of information storage and processing as well as cellular processes and signaling were significantly enriched in SFRFG. The flavor characteristics of FFRFG were mainly sourness as well as rich in sulfur organic compounds and broad range compounds, while that of SFRFG was mainly saltiness, umami, and bitterness, as well as rich in aromatic compounds. The findings could contribute to a comprehensive understanding of the role of FG in LFB fermentation from the perspective of rounds and depths.

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.

Metagenomic and physicochemical analyses reveal microbial community and functional differences between three types of low-temperature Daqu.

Complex microbes of different types of low-temperature Daqu (LTD) play an important role in the formation of flavors and qualities of light-flavor Baijiu during fermentation. However, characterizing the taxonomic and functional diversity of microbiota in three types of LTD (Houhuo, Hongxin, Qingcha) remains a major challenge. The present study combined metagenomic sequencing with culture-based methods and physicochemical analysis to compare the three LTD microbiota and elucidate their function in LFB brewing. The results revealed a high diversity of microbes in LTD, with 1286 genera and 4157 species detected across all studied samples. Bacteria and fungi were the main microbes in LTD, with a bacterial to fungal relative abundance ratio of above 4:1. Bacillus (21.18%) and Bacillus licheniformis (17.45%) were the most abundant microbes in the LTD microbiota at the genus and species levels, respectively. Culture-dependent analysis found the highest abundances of bacteria, fungi, and lactic acid bacteria in Houhuo, while the metagenomic-based microbiota found that the relative abundance of bacteria and fungi were highest in Houhuo and Hongxin among the three types of LTD, respectively. The different production temperatures of LTD had little effect on its microbial variety, but obviously impacted the microbiota structure and metagenomic function of LTD. Although the microbiota of the three types of LTD shared a high commonality, each had specific microbiota and functional metagenomic features, suggesting their different but complementary roles in the LFB fermentation process. The representative dominant microbes in Houhuo were mostly involved in metabolic pathways associated with the production of flavor substances in liquor. In contrast, the enriched microbes in Qingcha and Hongxin were not only capable of producing specific flavor substances but also had a strong ability to degrade macromolecular substances in raw materials, promoting microbial growth. This study has greatly enriched our knowledge of the effect of LTD fermentation temperature on its quality, providing practical and interesting information for future improvement of LTD and light-flavor Baijiu products.

Insights into the Aroma Profile of Sauce-Flavor Baijiu by GC-IMS Combined with Multivariate Statistical Analysis.

Aroma is among the principal quality indicators for evaluating Baijiu. The aroma profiles of sauce-flavor Baijiu produced by 10 different manufacturers were determined by GC-IMS. The results showed that GC-IMS could effectively separate the volatile compounds in Baijiu, and a total of 80 consensus volatile compounds were rapidly detected from all samples, among which 29 volatile compounds were identified, including 5 alcohols, 14 esters, 2 acids, 2 ketones, 5 aldehydes, and 1 furan. According to the differences in aroma profile found by multivariate statistical analysis, these sauce-flavor Baijiu produced by 10 different manufacturers can be further divided into three types. The relative odor activity value of the identified volatile compounds indicated that seven volatile compounds contributed most to the aroma of sauce-flavor Baijiu in order of aroma contribution rate, and they were ethyl hexanoate, ethyl pentanoate, ethyl 2-methylbutanoate, ethyl octanoate (also known as octanoic acid ethyl ester), ethyl 3-methylbutanoate, ethyl butanoate, and ethyl isobutyrate. Correspondingly, the main aromas of these sauce-flavor Baijiu produced by 10 different manufacturers were sweet, fruity, alcoholic, etheral, cognac, rummy, and winey. On the one hand, this study proved that GC-IMS is well adapted to the detection of characteristic volatile aroma compounds and trace compounds in Baijiu, which is of positive significance for improving the aroma fingerprint and database of sauce-flavor Baijiu. On the other hand, it also enriched our knowledge of Baijiu and provided references for the evaluation and regulation of the flavor quality of sauce-flavor Baijiu.

Evaluation of Bacterial Diversity and Evolutionary Dynamics of Gut Bifidobacterium longum Isolates Obtained from Older Individuals in Hubei Province, China.

Bifidobacterium longum predominates in the human gut throughout the life span, from birth to old age, and could alter the intestinal microbial population and immune function in the elderly. We investigated the intestinal bacterial diversity in the elderly, and further evaluated the genetic diversity and population structure of B. longum. The results revealed a distinct difference in gut bacterial populations between the elderly from Xiangyang and its neighboring region, Enshi city. A total of 62 bifidobacterial strains were isolated, 30 of which were found to be B. longum. The multilocus sequence typing (MLST) analysis also revealed that 437 B. longum isolates from diverse regions worldwide, including the 30 isolated in this study, could be classified into 341 sequence types (STs). They could be further clustered into 10 clonal complexes and 127 singleton STs, indicating a highly genetic diversity among B. longum isolates. Two putative clone complexes (CCs) containing the isolates from Xiangyang were found to be geographically specific, and a 213-bp recombination fragment was detected. Phylogenetic trees divided these 437 isolates into three lineages, corresponding to the three subspecies of B. longum. It is noteworthy that two isolates from the elderly were identified to be B. longum subsp. suis, while the others were B. longum subsp. longum. Together, our study characterized the intestinal bacterial diversity and evolution of B. longum in the elderly, and it could contribute to further studies on the genotyping and discrimination of B. longum. IMPORTANCE Bifidobacterium longum are common inhabitants of the human gut throughout the life span, and have been associated with health-promoting effects, yet little is known about the genotype profile and evolution of these isolates. Our study showed that there was significant difference in gut bacterial community and abundance of B. longum between the elderly from two neighboring cities. Furthermore, the possible geographically specific STs, CCs, and intraspecies recombination fragment were found among the B. longum isolates from elderly.

Diversity of microbiota, microbial functions, and flavor in different types of low-temperature Daqu.

Light-flavor Baijiu is made from grain materials using a combination of three types of low-temperature Daqu (Hongxin, Houhuo, and Qingcha). This study comprehensively examined the microbial structure, microbial functions, and flavor characteristics of the three types of low-temperature Daqu using high-throughput sequencing and electronic senses, and it further clarified the relationship between the microbiota and flavor in low-temperature Daqu. The results showed that Hongxin had the highest bacterial richness and diversity, while Houhuo had the lowest. Both fungal richness and diversity were significantly higher in Qingcha than in Hongxin and Houhuo. The differences in peak temperature during Daqu-making led to significant differences in the structure of microbial communities, microbial functions, and flavor quality in the three types of low-temperature Daqu, and could be leveraged for screening and enriching functional microorganisms for Baijiu-making. Co-exclusion patterns between lactic acid bacteria and Bacillus in low-temperature Daqu resulted in a negative correlation between amino acid transport metabolism and carbohydrate transport metabolism. The different types of low-temperature Daqu had distinct flavor profiles, and the differences in the taste profiles were more significant. Dominated by Thermoactinomyces and Lactobacillus, and together with Saccharopolyspora, Bacillus, Streptomyces, Saccharomycopsis, and Thermoascus, they formed the core microbiota that influencing the flavor of low-temperature Daqu. The bacteria mainly influenced the taste of low-temperature Daqu, whereas the fungi mainly influenced the aroma. Each type of low-temperature Daqu contributed to the flavor of light-flavor Baijiu: Hongxin could elevate the levels of aromatic compounds, Houhuo could regulate the bitterness and sourness, and Qingcha could inhibit the generation of sulfur organic compounds. The results of the present study enrich and refine our knowledge of low-temperature Daqu, promoting the further evolution of traditional brewing methods.

Comparative Analysis of Fecal Bacterial Microbiota of Six Bird Species.

The gut microbiota contributes to host health by maintaining homeostasis and improving digestive efficiency. Therefore, identifying gut microbes will shed light on the annual life cycle of animals and in particular those that are threatened or endangered. Nonetheless, the gut microbial composition of the majority of bird species is still unknown. Here, for the first time, 16S rRNA gene sequencing was used to characterize and compare the community composition and diversity of gut microbiotas from six species of birds raised at the Wildlife Conservation Center in Baotou, China: relict gull (Larus relictus; n = 3), muscovy duck (Cairina moschata; n = 3), ruddy shelduck (Tadorna ferruginea; n = 3), demoiselle crane (Anthropoides virgo; n = 4), whooper swan (Cygnus cygnus; n = 3), and black swan (Cygnus atratus; n = 5). A total of 26,616 operational taxonomic units from 21 samples were classified into 32 phyla and 507 genera. Chao1, Shannon diversity, observed species, and Simpson index analysis revealed differences in the community richness and diversity between the different species. Proteobacteria was the dominant bacterial phylum in whooper swan and relict gull, whereas Firmicutes was the dominant bacterial phylum in the other species. At the genus level, 11 dominant genera were detected (Lactobacillus, Psychrobacter, Enterococcus, Carnobacterium, Weissella, Burkholderia, Escherichia/Shigella, Leuconostoc, Buttiauxella, Desemzia, and Staphylococcus). Principal component and cluster analyses revealed that, while the microbial community composition of different individuals of the same species clustered together, the gut microbial composition varied between the bird species. Furthermore, the most abundant bacterial species differed between bird species. Because many avian gut microbes are derived from the diet, the eating habits and natural living environment of birds may be important contributing factors to the observed differences. Short-term changes to the diet and living environment have little effect on the composition of the avian gut microbiota. This study provides a theoretical basis for bird protection, including disease prevention and control.

Rice varieties affect bacterial diversity, flavor, and metabolites of zha-chili.

The structure and diversity of bacterial communities in spontaneously fermented zha-chili prepared using two different rice varieties (glutinous rice and indica rice) were investigated using high-throughput sequencing. Through metabolic pathway prediction, electronic senses and metabolite analysis, the relationships among the rice varieties used for preparation and the bacterial microbiota, flavor, and organic acid/amino acid metabolites in zha-chili were elucidated. We observed that the structure of bacterial communities in zha-chili samples differed significantly with the rice variety used during fermentation (p < 0.05), and that there was a greater abundance of bacterial species in zha-chili prepared using glutinous rice. Lactic acid bacteria were predominant in zha-chili, with an average relative abundance of 77.09%. The aroma of zha-chili was influenced by the raw material itself, while the characteristic tastes of zha-chili - including sourness, umami and richness - were significantly correlated with the bacterial microbiota. In addition, the abundance of lactic acid bacteria was positively correlated with the levels of organic acids and negatively correlated with the levels of amino acids. This also made the zha-chili prepared using glutinous rice sourer and imparted more umami taste to the zha-chili prepared using indica rice. Our observations provide a reference for the evaluation of zha-chili quality and could effectively guide the improvement of zha-chili products.

Characterization of bacterial community and flavor differences of different types of Douchi.

According to the appearance and technology, traditional fermented Douchi can be divided into dried Douchi and wet Douchi. However, there are few reports on the difference of bacterial community structure between them or the influence of bacterial community on product flavor. In this study, high-throughput sequencing technology and electronic nose were used to measure the bacterial diversity and flavor of 40 Douchi samples, and the correlation between them was explored by multivariate statistical means combined with COG database. Results showed that the cumulative average relative abundance of Firmicutes and Proteobacteria in the samples was as high as 95.93%, and the former was the core bacteria phylum. On the whole, the dominant bacteria in Douchi were Bacillus (50.67%), Staphylococcus (14.07%), Enterococcus (2.54%), Proteus (1.61%), Brevibacillus (1.46%), Providencia (1.26%), Weissella (1.24%), and Ureibacillus (1.19%). LEfSe analysis indicated that Bacillus can be used as a biomarker in dried fermented soybeans. Meanwhile, dried samples contained more intensive aromatic substances, but were significantly lower in W6S (selectivity to hydrogen) and W3S (methane-aliph) compared with the wet samples. Aneurinibacillus and Brevibacillus were helpful to the formation of aromatic flavor in Douchi, but Vagococcus and Corynebacterium were the opposite. Gene and microbial phenotypic prediction showed that microorganisms in dried Douchi use protein more efficiently, while in wet Douchi, microbial energy metabolism was more vigorous. The pathogenic potential of microorganisms in dried samples was higher than that in wet. This study can sound the alarm for improving the safety of home-brewed Douchi and provide guidance for the subsequent screening of strains that enhance the flavor of fermented soybeans.

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.

Compositional and genetic alterations in Graves' disease gut microbiome reveal specific diagnostic biomarkers.

Graves' Disease is the most common organ-specific autoimmune disease and has been linked in small pilot studies to taxonomic markers within the gut microbiome. Important limitations of this work include small sample sizes and low-resolution taxonomic markers. Accordingly, we studied 162 gut microbiomes of mild and severe Graves' disease (GD) patients and healthy controls. Taxonomic and functional analyses based on metagenome-assembled genomes (MAGs) and MAG-annotated genes, together with predicted metabolic functions and metabolite profiles, revealed a well-defined network of MAGs, genes and clinical indexes separating healthy from GD subjects. A supervised classification model identified a combination of biomarkers including microbial species, MAGs, genes and SNPs, with predictive power superior to models from any single biomarker type (AUC = 0.98). Global, cross-disease multi-cohort analysis of gut microbiomes revealed high specificity of these GD biomarkers, notably discriminating against Parkinson's Disease, and suggesting that non-invasive stool-based diagnostics will be useful for these diseases.

Using PacBio sequencing to investigate the effects of treatment with lactic acid bacteria or antibiotics on cow endometritis

BACKGROUND: Endometritis is the most common disease of dairy cows and traditionally treated with antibiotics. Lactic acid bacteria can inhibit the growth of pathogens and also have potential for treatment of endometritis. Using PacBio single-molecule real-time sequencing technology, we sequenced the fulllength l6S rRNA of the microbiota in uterine mucus samples from 31 cows with endometritis, treated with lactic acid bacteria (experimental [E] group) and antibiotics (control [C] group) separately. Microbiota profiles taken before and after treatment were compared. RESULTS: After both treatments, bacterial species richness was significantly higher than before, but there was no significant difference in bacterial diversity. Abundance of some bacteria increased after both lactic acid bacteria and antibiotic treatment: Lactobacillus helveticus, Lactococcus lactis, Lactococcus raffinolactis, Pseudomonas alcaligenes and Pseudomonas veronii. The bacterial species that significantly decreased in abundance varied depending on whether the cows had been treated with lactic acid bacteria or antibiotics. Abundance of Staphylococcus equorum and Treponema brennaborense increased after lactic acid bacteria treatment but decreased after antibiotic treatment. According to COG-based functional metagenomic predictions, 384 functional proteins were significantly differently expressed after treatment. E and C group protein expression pathways were significantly higher than before treatment (p < 0.05). CONCLUSIONS: In this study, we found that lactic acid bacteria could cure endometritis and restore a normal physiological state, while avoiding the disadvantages of antibiotic treatment, such as the reductions in abundance of beneficial microbiota. This suggests that lactic acid bacteria treatment has potential as an alternative to antibiotics in the treatment of endometritis in cattle.

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.

Analysis of microbial diversity and functional differences in different types of high-temperature Daqu.

Bacterial communities that enrich in high-temperature Daqu are important for the Chinese maotai-flavor liquor brewing process. However, the bacterial communities in three different types of high-temperature Daqu (white Daqu, black Daqu, and yellow Daqu) are still undercharacterized. In this study, the bacterial diversity of three different types of high-temperature Daqu was investigated using Illumina MiSeq high-throughput sequencing. The bacterial community of high-temperature Daqu is mainly composed of thermophilic bacteria, and seven bacterial phyla along with 262 bacterial genera were identified in all 30 high-temperature Daqu samples. Firmicutes, Actinobacteria, Proteobacteria, and Acidobacteria were the dominant bacterial phyla in high-temperature Daqu samples, while Thermoactinomyces, Staphylococcus, Lentibacillus, Bacillus, Kroppenstedtia, Saccharopolyspora, Streptomyces, and Brevibacterium were the dominant bacterial genera. The bacterial community structure of three different types of high-temperature Daqu was significantly different (p < .05). In addition, the results of microbiome phenotype prediction by BugBase and bacterial functional potential prediction using PICRUSt show that bacteria from different types of high-temperature Daqu have similar functions as well as phenotypes, and bacteria in high-temperature Daqu have vigorous metabolism in the transport and decomposition of amino acids and carbohydrates. These results offer a reference for the comprehensive understanding of bacterial diversity of high-temperature Daqu.

The Depth-Depended Fungal Diversity and Non-depth-Depended Aroma Profiles of Pit Mud for Strong-Flavor Baijiu.

Microorganisms in pit mud are the essential factor determining the style of strong flavor Baijiu. The spatial distribution characteristics of fungal communities and aroma in the pit mud for strong flavor Baijiu from Xinjiang, China, were investigated using Illumina MiSeq high-throughput sequencing and electronic nose technology. A total of 138 fungal genera affiliated with 10 fungal phyla were identified from 27 pit mud samples; of these, Saccharomycopsis, Aspergillus, and Apiotrichum were the core fungal communities, and Aspergillus and Apiotrichum were the hubs that maintain the structural stability of fungal communities in pit mud. The fungal richness and diversity, as well as aroma of pit mud, showed no significant spatial heterogeneity, but divergences in pit mud at different depths were mainly in pH, total acid, and high abundance fungi. Moisture, NH4 +, and lactate were the main physicochemical factors involved in the maintenance of fungal stability and quality in pit mud, whereas pH had only a weak effect on fungi in pit mud. In addition, the fungal communities of pit mud were not significantly associated with the aroma. The results of this study provide a foundation for exploring the functional microorganisms and dissecting the brewing mechanism of strong flavor Baijiu in Xinjiang, and also contributes to the improvement of pit mud quality by bioaugmentation and controlling environmental physicochemical factors.

Bacterial Communities and Prediction of Microbial Metabolic Pathway in Rice Wine Koji From Different Regions in China.

Rice wine koji, a traditional homemade starter culture in China, is nutritious and delicious. The final quality of rice wine koji is closely related to the structure of its microbial community. However, the diversity of natural microorganisms in rice wine koji from different regions has not been evaluated. In this study, the microbial population of 92 naturally fermented rice koji samples collected from Hubei, Guangxi, and Sichuan was systematically analyzed by high-throughput sequencing. From all the rice wine koji samples, 22 phyla and 479 bacterial genera were identified. Weissella, Pediococcus, Lactobacillus, Enterobacter, Lactococcus, Pantoea, Bacillus, Staphylococcus, and Leuconostoc were the dominant genera in rice wine koji. The bacterial community structure of rice wine koji samples from different regions was significantly different (p < 0.05). The bacterial community composition of the samples from Hubei and Guangxi was similar, but significantly different from that of SC samples (p < 0.05). These differences may be caused by variations in geography, environment, or manufacturing. In addition, the results of microbial phenotype prediction by BugBase and bacterial functional potential prediction by PICRUSt showed that eight of the nine predicted phenotypic functions of rice wine koji samples from different regions were significantly different (p < 0.05) and that vigorous bacterial metabolism occurred in rice wine koji samples.