Comparative analysis of sensory features, microbial diversity, and their correlations in light-flavor Daqu from different regions.

This study performed a comparative analysis of the sensory and microbial profiles of light-flavor Bijou (LFD) from Taiyuan (Shanxi Province) and Suizhou (Hubei Province) in China. The results of the electronic nose showed that the aromatic substances of the LFD from Taiyuan (TLFD) were significantly higher (p < .05), while alcohol and aldehyde substances were significantly lower (p < .05) compared with the LFD from Suizhou (SLFD). The average response values of sensors W1C (sensitive to aromatic hydrocarbons), W3C (sensitive to amine and aromatic components), W5C (sensitive to olefins, aromatics, and polar molecules), and W2S (sensitive to alcohol and aldehyde compounds) to TLFD were 0.26, 0.33, 0.34, and 7.72, whereas the response values to SLFD were 0.25, 0.32, 0.33, and 8.04, respectively. The electronic tongue results showed that the aftertaste A (bitter aftertaste) and aftertaste B (astringent aftertaste) of the TLFD were significantly higher (p < .05) and umami was significantly lower (p < .05) as compared to the SLFD. The relative intensities of the aftertaste A, aftertaste B, and umami indicators of TLFD were 0.10, -0.008, and -0.22, respectively, while those of SLFD were -0.23, -0.36, and 0.835, respectively. MiSeq high-throughput sequencing results showed that TLFD exhibited lower fungal richness and diversity compared to SLFD. The dominant bacterial genera were mainly Bacillus (58.12%), Kroppenstedtia (10.11%), and Weissella (6.26%), and the dominant fungal genera were Saccharomycopsis (67.53%), Rasamsonia (9.90%), and Thermoascus (7.10%). Streptomyces and Staphylococcus were identified as the key characteristic microorganisms in TLFD, while Kroppenstedtia, Rasamsonia, and Thermoascus were the key characteristic microorganisms in SLFD. Correlation analysis indicated a stronger correlation between microorganisms and sensory characteristics in SLFD samples. This study provides valuable insights into the sensory and microbiological characteristics of LFD from different regions and offers a new perspective for understanding the production of differently flavored light-flavor Baijiu.

Comparative genomic analysis of the genus Weissella and taxonomic study of Weissella fangxianensis sp. nov., isolated from Chinese rice wine starter.

In this study, three lactic acid bacteria, namely, HBUAS51963T, HBUAS51964 and HBUAS51965, were isolated from Chinese rice wine starter sampled in Fangxian County, PR China. All were non-motile, non-spore-forming and Gram-positive spherical cells. Their taxonomic status was characterized using a polyphasic approach. Genome-based analysis revealed that all three strains were phylogenomically related to Weissella thailandensis KCTC 3751T and Weissella paramesenteroides ATCC 33313T. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between the three strains and the phylogenetically related type strains were less than 54.8 and 93.8 %, respectively, and thus, they were below the thresholds of dDDH and ANI for species definition. The genomic DNA G+C content was 38.6 mol %. The predominant fatty acid methyl esters (>10 %) were C16 : 0, C19 : 0 cyc11 and summed feature 10 (C18 : 1 cyc11 and/or ECL 17.834). The polar lipids in the cells of strain HBUAS51963T were mainly phosphatidylglycerol, diphosphatidylglycerol, unidentified glycolipids, phospholipids and lipids. Finally, the three strains were capable of producing d-lactic acid (4.29 g l-1) and various organic acids such as tartaric, acetic, lactic and succinic acids. Overall, the results of genotypic, phenotypic and genomic analyses suggest that the three strains represent a new species of the genus Weissella, for which the name Weissella fangxianis sp. nov. is proposed. The type strain is HBUAS51963T (=GDMCC 1.3506T= JCM 35803T).

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.

Characterization of two novel pentose-fermenting and GABA-producing species: Levilactobacillus tujiorum sp. nov. and Secundilactobacillus angelensis sp. nov. Isolated from a solid-state fermented zha-chili.

Lactobacilli are dominant in zha-chili. This study provides a taxonomic characterization of five bacterial strains isolated from zha-chili in China. The cells were Gram-positive, facultative anaerobic, non-spore-forming, flagella-free, catalase-negative, heterofermentative, pentose-fermenting, and gamma-aminobutyric acid (GABA)-producing rods. For HBUAS51241T, HBUAS51329, and HBUAS51416, C16:0, C18:1ω9c and C19:0 iso were the predominant cellular fatty acids; diphosphatidylglycerol (DPG), phosphatidylglycerol (DP), glycolipids (GL), and glycolipids (AL) were the major phospholipids. While for HBUAS51383T and HBUAS58055, C16:0, C18:1ω9c, C19:0 cyclo ω8c were the predominant cellular fatty acids; DPG, DP, GL, and AL were the major phospholipids. Strains HBUAS51241T, HBUAS51329, and HBUAS51416 showed 98.1-99.1% 16S rRNA gene sequence similarity, 80.2-81.4% ANI, 87.7-90.0% AAI, and 23.8-32.8% digital DDH to their closest related type strains Levilactobacillus hammesii DSM 16381T, Levilactobacillus parabrevis ATCC 53295T, and Levilactobacillus fuyuanensis 244-4T. Strains HBUAS51383T and HBUAS58055 showed 98.7-99.5% 16S rRNA gene sequence similarity, 75.4-81.4% ANI, 75.5-89.1% AAI, and 19.7-24.0% digital DDH to their closest related type strains Secundilactobacillus silagincola IWT5T, Secundilactobacillus silagei JCM 19001T, Secundilactobacillus pentosiphilus IWT25T, Secundilactobacillus mixtipabuli IWT30T, Secundilactobacillus odoratitofui DSM 19909T, and Secundilactobacillus similis DSM 23365T. The central carbon metabolism pathways for the five strains were summarizeded. Based on the phenotypic, chemotaxonomic, and genomic data, we propose two novel species Levilactobacillus tujiorum sp. nov. whose type strain is HBUAS51241T (=GDMCC 1.3022T = JCM 35241T), and Secundilactobacillus angelensis sp. nov. whose type strain is HBUAS51383T (=GDMCC 1.3021T = JCM 35209T).

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.

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.

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.

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.

High-throughput sequencing-based analysis of fungal diversity and taste quality evaluation of Douchi, a traditional fermented food.

Douchi, a popular traditional fermented soybean product, is mainly made by natural fermentation. However, its taste quality is affected by specific fungal communities which vary greatly according to fermentation conditions and production technologies used in different regions. Therefore, the taste quality of Douchi samples from different regions was digitally evaluated using electronic tongue technology. In addition, the fungal community structures and its association of them were also identified using high-throughput sequencing technology. Results showed that there were obvious differences in the taste quality of samples from different regions, while the tastes of different types of samples from the same region were similar. Sourness, umami, richness, and saltiness were the main reasons for regional differences in taste. Similarly, the results of high-throughput sequencing indicated that samples from different regions displayed important differences in dominant fungal genus, among which Debaryomyces, Fusarium, Pichia, Aspergillus, and Saccharomyces were the main dominant fungi. Debaryomyces and Trichosporon were conducive to the formation of taste qualities of Douchi, while Cladosporium and Candida have a negative impact on the taste quality of Douchi var correlation analysis. This study indicated the effects of dominant fungi on the formation of Douchi taste quality, allowing a deeper understanding of the role of microbial species in generating fermented soybean products in China. Our work provides theoretical support to guide the improvement of the industrial production process of Douchi.