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In recent years, the incidence of female infertility has risen sharply, which is affected by many factors. It was recognized that female reproductive tract microbes play a role in the process of female conception. If the reproductive tract microbes could solve a certain proportion of infertility, it would certainly reduce the pain and economic burden of many patients. The objective of this study was to investigate the microbial community composition of follicular fluid in infertile patients and its potential impact on infertility.Follicular fluid from 49 primary infertility and 52 secondary infertility patients was collected by a negative pressure needle, and the microbiota was analyzed by 16S rDNA sequencing.It was found that Lactobacillus, especially L. crispatus, might have a positive effect on female pregnancy. Considering the presence or absence of male factors and different body mass indices, L. iners might inhibit female pregnancy. However, L. iners seemed to play a positive role in egg maturation, while Gardnerella and Cutibacterium acnes might have a negative effect on female pregnancy.This study suggested the potential role of Lactobacillus in follicular fluid in improving female infertility and provided a theoretical basis for the future microbiological treatment of female infertility.

Lactobacillus plantarum SP8, isolated from traditional Chinese pickle juice, was utilized for the production of exopolysaccharides (EPSs), but the EPS yield was low under normal MRS medium. The single factor experiment and response surface methodology were used to optimize the medium components and culture conditions and the optimal conditions for EPS production were successfully obtained. Results showed that the optimum condition was glucose 22 g/L, yeast extract 30 g/L, fermentation temperature 35.6 °C, fermentation time 22 h and the theoretical EPS yield was 282.494 mg/L. The results were similar to the actual yield, 280.105 mg/L. By optimizing the culture conditions, the yield of L. plantarum SP8 EPS was improved by nearly 19 times. In the gas chromatography analysis, it was found that L. plantarum SP8 EPS consisted of d-rhamnose, arabinose, galactose, and d-acetylglucosamine, but glucose was not included, which was quite different from the reported heteropolysaccharide component of Lactobacillus. Furthermore, the antioxidant activity of L. plantarum SP8 EPS was evaluated with the in vitro scavenging abilities on DPPH·, $${\text{O}}_{2}^{ - }$$ and ·OH. The in vitro antioxidant activity study indicated that L. plantarum SP8 EPS possessed certain antioxidant activity. All results demonstrated the potential of L. plantarum SP8 in the food and dairy industry.

Traditional fermented soybean contains a variety of microflora. To obtain a comprehensive and accurate understanding of the microbial community structure and metabolic potential of fermented soybean, comparative metagenomics and metatranscriptomics were performed on a sample of fermented soybean in Yunnan Province. Metagenomic DNA and metatranscriptomic RNA were sequenced using Illumina HiseqTM2500, which yielded a total of 92,192,276 reads, with an average read length of 150 bp and 38,798,262 paired-end sequences, with an average length of 151 bp. The results show that Providencia stuartii was the most abundant species and the genes of carbohydrates (2296, 13.30%), protein metabolism (1530, 8.86%), and amino acids and amino acid derivatives (1423, 8.24%) were dominant. The expression levels of the genes belonging to amino acid transport and metabolism processes were the highest according to the reads per length of transcript in kilo-bases per million mapped reads (RPKM) values, followed by energy production and conversion and carbohydrate transport and metabolism. The metabolic pathways were primarily associated with carbohydrates, proteins and amino acids, which might be in accordance with the high levels of proteins and other nutrients in soybeans. Overall, these findings provide insights into the community structure and metabolic potential of the fermented soybean microbiome.

The goal of this study was to explore the regulatory mechanisms of phenyllactic acid (PLA) overaccumulation in Lactobacillus plantarum. The dynamics of PLA production revealed that 24 h was a suitable fermentation time, at which one of the largest differences in PLA content between strains S1 and YM-4-3 was 22.42 mg/L. Additionally, an optimization experiment showed that PLA production under the optimal condition (sample YM-4-3y) was up to 400.74 mg/L, 7.61-13.26 times as those of YM-4-3 and S1. Subsequently, an integrated analysis of genomic, transcriptomic and metabolomic data revealed that, YM-4-3 and YM-4-3y, compared with S1, although lacking a complete de novo biosynthetic pathway, increased PLA production by strengthening the core pathway and central carbon metabolism, and weakening the biosynthesis pathway of amino acids and their derivatives. These changes can provide sufficient precursors and compensate for or balance the energy consumed by the reinforced core pathway.

This work aims to uncover how glucose affected the production of phenyllactic acid (PLA) and p-hydroxyphenyllactic acid ( p-OH-PLA). The highest yields of PLA (68.53 mg/L) and p-OH-PLA (50.39 mg/L) were observed after Lactobacillus plantarum strain YM-4-3 fermentation in media containing 30 and 10 g/L glucose, respectively. Additionally, the antimicrobial activity of YM-4-3 against food-borne pathogens and the NADH/NAD+ ratio were positively correlated with the production of PLA and p-OH-PLA, respectively. In addition, a 2-oxoglutarate/malate translocator coding gene ( Omt1) was selected based on the qPCR results, and its knockout mutant, compared with the wild-type strain YM-4-3, showed that the PLA and p-OH-PLA production was decreased by 1.37-6.99 and 1.53-1.59 times, respectively. This result indicated that OMT1 was involved in the biosynthesis of PLA and p-OH-PLA. To conclude, this study suggests that glucose, NADH/NAD+ ratio and/or the Omt1 gene, PLA, and p-OH-PLA production, and antimicrobial activity contribute to a cause-and-effect relationship.

Lactobacillus (Lb.) plantarum is typically used as a starter culture in salt-fermented foods. Here we report 3-4% NaCl reducing the antimicrobial activity of Lb. plantarum strain YM-4-3, owing to the decrease of bacterial growth, plantaricin activity and expression levels of plantaricin biosynthesis-related genes (PBGs). Meanwhile, 1% NaCl promoted slightly the growth of YM-4-3 and up-regulated the expression of PBGs to the greatest level. The results from a spoilage experiment of fermented soybean products revealed that the 1% NaCl and YM-4-3 treatment group had the longest shelf life representing the minimum number of pathogenic bacteria and the lowest degree of mildew. Therefore, a combination of Lb. plantarum with a low concentration of salt, such as 1% NaCl, is a recommended condition for preparing fermented foods.

Intestinal bacteria are symbiotic microbiota within the human gut and are implicated in the occurrence and development of colorectal cancer (CRC). The current study investigated the changes in bacterial composition prior to and following surgery, as well as the differences in the bacterial community structure between cancer tissue and adjacent normal tissue. The diversity of the bacterial community and the composition of the bacteria were assessed. In addition, phylogenetic analysis and principle component analysis (PCA) were performed. The results revealed that cancer tissue and adjacent normal tissue exhibited similar bacterial compositions. However, a significant difference was identified in the composition of intestinal bacteria in stool samples collected from patients following surgery compared with stool samples collected prior to surgery. Each patient had their own unique intestinal bacterial community, likely due to a number of factors, including diet, genetic factors and health status. In addition, phylogenetic trees revealed that the most abundant operational taxonomic unit, 0001, was associated with Escherichia coli in all samples. Finally, PCA suggested that the bacterial community structure in all patient stools was similar following surgery. The current study provides information regarding the diversity of the intestinal bacterial community of patients with CRC and provides a basis for postoperative intestinal assessments.

Tobacco brown spot, caused by Alternaria species, is a devastating tobacco disease. To explore the role of a group III histidine kinase (AlHK1) on A. longipes pathogenesis, the invasion progress of A. longipes was monitored. We found that the wild-type strain C-00 invaded faster than the AlHK1-disrupted strain HK∆4 in the early and middle infection stages and the reverse trend occurred in the late infection stage. Then, eight invasion transcriptomes were performed using RNA-Seq and 205 shared, 505 C-00 and 222 HK∆4 specific differentially expressed genes (DEGs) were identified. The annotation results showed seven antioxidant activity genes were specifically identified in the HKΔ4 DEGs. A subsequent experiment confirmed that HKΔ4 was more resistant to low concentrations oxidative stress than C-00. In addition, the results from 1) statistics for the number of DEGs, GO enriched terms, DEGs in clusters with rising trends, and 2) analyses of the expression patterns of some DEGs relevant for osmoadaptation and virulence showed that changes in C-00 infection existed mainly in the early and middle stages, while HKΔ4 infection arose mainly in the late stage. Our results reveal firstly the pathogenesis of A. longipes regulated by AlHK1 and provide useful insights into the fungal-plant interactions.

Lactobacillus plantarum SP8, isolated from traditional Chinese pickle juice, was utilized for the production of exopolysaccharides (EPSs), but the EPS yield was low under normal MRS medium. The single factor experiment and response surface methodology were used to optimize the medium components and culture conditions and the optimal conditions for EPS production were successfully obtained. Results showed that the optimum condition was glucose 22 g/L, yeast extract 30 g/L, fermentation temperature 35.6 °C, fermentation time 22 h and the theoretical EPS yield was 282.494 mg/L. The results were similar to the actual yield, 280.105 mg/L. By optimizing the culture conditions, the yield of L. plantarum SP8 EPS was improved by nearly 19 times. In the gas chromatography analysis, it was found that L. plantarum SP8 EPS consisted of d-rhamnose, arabinose, galactose, and d-acetylglucosamine, but glucose was not included, which was quite different from the reported heteropolysaccharide component of Lactobacillus. Furthermore, the antioxidant activity of L. plantarum SP8 EPS was evaluated with the in vitro scavenging abilities on DPPH·, [Formula: see text] and ·OH. The in vitro antioxidant activity study indicated that L. plantarum SP8 EPS possessed certain antioxidant activity. All results demonstrated the potential of L. plantarum SP8 in the food and dairy industry.

Folate is an important intermediate in cellular metabolism. However, because of a lack of key enzymes in the folate biosynthetic pathway, humans require supplementation with dietary folate. Some Lactobacillus plantarum strains have the ability to produce folate. To gain a better understanding of the folate biosynthetic pathway in the L. plantarum strain 4_3, which generates high folate yields, L. plantarum strain 4_3 was grown in folic acid casei medium (FACM) and fermented soybean, after obtaining a draft genome sequence. The pH values and folate yields were monitored during culturing, as were the transcriptomic profiles of cultured bacteria. The folate content increased for 12 h and then decreased before increasing again. All the genes involved in the de novo biosynthesis of folate were detected in both the genomic and transcriptomic data. The upregulation of the para-aminobenzoate biosynthesis pathway could explain the folate production in fermented soybean. Soybeans are a good substrate for the production of functional foods because of their well-suited cultivation and nutritional quality. The results of this study provide a good explanation for the high folate production observed during the fermentation of soybeans.

The protein kinase HOG1 in high osmolarity glycerol pathway is one of mitogen-activated protein kinases and plays an important role in fungi cellular responses to external stress and stimuli. In this study, AlHOG1, a yeast HOG1 homolog, was isolated from tobacco pathogenic fungus Alternaria longipes and its disrupted mutants were generated using homologous recombination. Compared with the wild-type strain, the mutants grew slower under Minimal-Media, produced less conidia on Potato-Dextrose-Agar and showed a retardation phenotype in conidial germination rate. Colony color observation indicated that AlHOG1 most likely negatively regulated melanin production under nutrients-deficient or osmotic stress. The same action mode of AlHOG1 on hyphal growth in response to certain carbon or nitrogen sources was speculated. Additionally, the disrupted strains were sensitive to osmotic and oxidative stress, and showed increased resistance to dicarboximide, phenylpyrrole and carbendazim fungicides. Meanwhile, results from the comparative analysis between the AlHOG1- and the AlHK1 (encoding a group III histidine kinase)-disrupted mutants showed that AlHOG1 mediated-pathway was partly regulated by the upstream AlHK1 kinase for cellular resistance to salts osmotic stress and that AlHK1 was a primary regulator for cellular sugar and fungicides stress adaptation and likely modulated both AlHOG1-mediated pathway and other unknown mechanisms. Noteworthily, AlHOG1 but not AlHK1 was confirmed to be associated with high concentration glycerol adaptation, which is the first report to our knowledge. Overall, our results indicated that AlHOG1 was involved in multiple physiological processes in A. longipes, including mycelial and conidial development, melanin production, nutrients sensing and multi-stress responses.

Background: Mothers are the primary source of bacteria for newborns, but it is unclear whether the structuring of the infant microbiome is the result of mother-to-newborn transmission prior, during or after birth. Similarly, the effect of delivery mode on neonatal microorganisms has been the focus of controversy. Methods: Approximately the same number of healthy maternal and neonatal pairs of vaginal birth and caesarean section were enrolled in this study. Meconium, placenta, membranes and amniotic fluid samples of newborns and vaginal, rectal and oral samples of mothers were collected. All samples were amplified and sequenced by a 16S rRNA gene primer set targeting bacteria and archaea. Findings: A total of 550 samples from 36 mother-neonate pairs with vaginal births and 42 mother-neonate pairs with caesarean sections were included in this study. A negative control showed that all data analysis in this study was not affected by contamination. Indeed, there was a high diversity of microbial communities in the pregnant environment of the foetus. The microbial community composition of neonatal samples was not affected by the mode of delivery. Approximately half of all meconium microorganisms were found in the vaginal, rectal and oral samples of the mother. Interpretation: The origin and influencing factors of pioneer microorganisms in neonates were studied. How these microorganisms enter the breeding environment and their roles need further study. Funding Statement: the National Natural Science Foundation of China (81860437) and Yunnan Province Innovation Team of Intestinal Microecology Related Disease Research and Technological Transformation (China). Declaration of Interests: The authors declare no competing interests. Ethics Approval Statement: This project was approved by the Ethics Committee of Yan’an affiliated Hospital of Kunming Medical University (2019-077-01). Mothers were made aware of the nature of the study, specifically consented to give their personal information, and gave written informed consent for their and their child’s participation.

Lactobacillus plantarum is an important probiotic and is mostly isolated from fermented foods. We sequenced the genome of L. plantarum strain 5-2, which was derived from fermented soybean isolated from Yunnan province, China. The strain was determined to contain 3114 genes. Fourteen complete insertion sequence (IS) elements were found in 5-2 chromosome. There were 24 DNA replication proteins and 76 DNA repair proteins in the 5-2 genome. Consistent with the classification of L. plantarum as a facultative heterofermentative lactobacillus, the 5-2 genome encodes key enzymes required for the EMP (Embden-Meyerhof-Parnas) and phosphoketolase (PK) pathways. Several components of the secretion machinery are found in the 5-2 genome, which was compared with L. plantarum ST-III, JDM1 and WCFS1. Most of the specific proteins in the four genomes appeared to be related to their prophage elements.

Yond bap is a traditional fermented milk product from the Yunnan province in China. Yond baps are mostly produced with goat’s milk and occasionally with cow’s milk. As a traditional dairy product produced by fermentation, yond bap contains an abundant microflora. The microbes that play a role in the fermentation of goat milk form a specific community structure. In this work, 11 types of yond bap were characterized from four different regions in the Yunnan province. We employed bar-coded pyrosequencing of 16S rRNA to analyze the bacterial community structure in the yond baps. The results indicated that the bacterial community structures of the 11 yond baps were different. Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were detected in all the samples. Pseudomonas, a proteobacterium, was the predominant bacterial genus. Pseudomonas sequences were detected often and in most of the samples. Next, Streptococcaceae, Lactobacillaceae, and Lactococcus belonging to Streptococcaceae family played important roles in the fermentation of yond baps as the numbers of sequences belonging to these species were higher. The results demonstrate the presence of different bacterial community structures of yond baps from different regions in the Yunnan Province of China. This study provides information about the diversity of the microbial community of fermented food and a reliable theoretical foundation for improving the quality and flavor of yond baps.

The gut microbiota is known to have a considerable impact on health; some types of organisms, including bifidobacteria and lactobacilli, are considered to be ‘good’ colonizing bacteria, while other species are potentially pathogenic. The initial colonization of a newborn infant affects his/her health in the future. Recently, research has indicated that the fetus may actually be colonized with bacteria during gestation. Accordingly, this study was conducted to assess the similarities in the bacterial species detected in the infant’s first feces with samples obtained from the mother’s placenta and vagina., BACKGROUND: The gut microbiota plays an important role in human health. It is essential to understand how the composition of the gut microbiota in neonates is established. OBJECTIVES: To investigate the nature of the microbial community in the first feces of newborn infants compared with the mothers’ placentae and vaginas. METHODS: One infant who was delivered via Cesarean section was compared with an infant who was delivered vaginally. Bar-coded pyro-sequencing of 16S ribosomal RNA genes was used to investigate the bacterial community composition and structure of each site. RESULTS: Neonatal feces of both infants had similar bacterial communities, and they were similar to the mother’s placenta regardless of the method of delivery. The vaginal bacterial community differed between the two mothers, but not different sites within the vagina. The bacteria in the neonatal feces and the mothers’ placentae demonstrated considerably higher diversity compared with the vaginas. The family Lactobacillaceae dominated in the vaginal samples, while the most abundant family in the fecal and placental samples was Micrococcineae. CONCLUSIONS: These results may provide new directions for the study of infant gut microbial formation.

Quorum sensing (QS) is a means of cell-to-cell communication that regulates, via small signalling molecules, expression of a series of genes and controls multicellular behaviour in many bacterial species. The enzyme S-ribosylhomocysteine lyase (LuxS) transforms S-ribosylhomocysteine (SRH) into 4, 5-dihydroxy-2, 3-pentanedione (DPD), the precursor of the interspecies QS signalling molecule autoinducer-2 (AI-2). In this study, two LuxS-coding genes, luxS1 and luxS2, with 70% sequence identity were isolated from Lactobacillus plantarum YM-4-3, and overexpressed in Escherichia coli BL21 (DE3), and the protein products were purified successfully. After incubation of LuxS1 or LuxS2 with SRH, the reaction products were able to induce Vibrio harveyi BB170 bioluminescence, clearly demonstrating that both LuxS1 and LuxS2 synthesize AI-2 from SRH in vitro. Ellman's assay results revealed optimal temperatures for LuxS1 and LuxS2 of 45 and 37 °C, respectively, and their activities were stimulated or inhibited by several metal ions and chemical reagents. In addition, enzyme kinetics data showed that Km, Vmax and Kcat value of LuxS1 for the substrate (SRH) were higher than that of LuxS2. These results suggest that LuxS1 and LuxS2 mediate QS in a temperature-dependent manner and may play conserved roles in AI-2 synthesis but exhibit different activities in response to external environmental stress. To our knowledge, this paper is the first report of two luxS genes present in one bacterial genome and the subsequent comparative elucidation of their functions in AI-2 production. Collectively, our study provides a solid basis for future research concerning the AI-2/LuxS QS system in L. plantarum YM-4-3.

Three new hydroquinone terpenoids with benzogeijerene skeletons, euchroquinols A-C (1- 3), and a new monoterpenylbenzenoid, 9,17-epoxyarnebinol (4), along with five known compounds were isolated from the stem bark of ARNEBIA EUCHROMA. Shikonin (6) exhibited potent anti-HCV activity with a selective index of 43.56, and compounds 1, 6, and des-O-methyllasiodiplodin (7) showed anti-Staphylococcus aureus activity with MICs of 0.5, 0.125, and 0.125 mg/mL, respectively.

Two hundred and fourteen endophytic fungi were isolated from 500 segments of aquatic/riparian plants Ottelia acuminata, Myriophyllum verticillatum, Equisetum arvense, Cardamine multijuga, and Impatiens chinensis. They were identified to 31 taxa in which Cladosporium, Fusarium, and Geotrichum were the dominant genera. Among all isolates, 169 (79%) were anamorphic fungi, 1 (0.5%) was an teleomorphic ascomycete and 44 (21%) were sterile mycelia. There were significant differences in the colonization frequency of endophytes between the five plant species (X∼2=51.128, P

A xylanase gene (xynE) encoding XynE (110 kDa) was cloned from a lambda phage genomic library of Aeromonas caviae ME-1 which is a multiple-xylanase-producing bacterium. Upon nucleotide sequence analysis, we found that xynE comprises 2823 by and encodes a protein of 941 amino acid residues (104,153 Da), which was similar to endo-beta-1,4-xylanases which are categorized to glycosyl hydrolase family 10. An Escherichia coli transformant that harbored pXED30 carrying xynE produced 110-, 84-, 72-, and 66-kDa xylanases in the cell-free extract, and 72- and 66-kDa xylanases in the culture supernatant. We purified the 66-kDa xylanase to electrophoretic homogeneity from a culture supernatant by a series of column chromatographies. The calculated molecular mass of the purified xylanase determined by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was 60,154.50 Da, and the xylanase was designated XynE60. Analysis of the N-terminal 10 amino acid residues and the determined molecular mass of XynE60 revealed that XynE60 is a product processed at the Gly26-Gly27, and Thr565-Ala566 sites of XynE by proteolytic cleavage. XynE60 showed optimal activity at 55 degrees C and pH 8.0, and was stable below 45 degrees C and at pH 7.0-8.5. The K(m) and V(max) of XynE60 were calculated to be 8.1 mg/ml and 6897 nkat/mg, respectively.

Lactobacillus plantarum is an important probiotic that is isolated mostly from fermented foods. Here, we report the first draft genome sequence of L. plantarum strain AY01, isolated from the raw material of fermented goat milk cheese. This bacterium, with optimum growth at 30°C, has a G+C content of 43.68%.

This research was aimed at isolating and identifying the predominant lactic acid bacteria (LAB) in the traditional Chinese salt-fermented soybean food, douchi, from Yunnan, China. The predominant LAB present were isolated and identified by conventional culture-dependent methods combined with molecular biological methods. Two hundred and sixty isolates were obtained from thirty kinds of traditional fermented douchi from six cities and counties in Yunnan, and those strains were divided into twelve groups by their morphological and biochemical characteristics. Based on 16S ribosomal DNA (rDNA) sequencing and analysis, 56 representative strains were identified as belonging to 6 genera and 14 species: Lactobacillus (4 spp.), Weissella (3 spp.), Pediococcus (2 spp.), Staphylococcus (2 spp.), Enterococcus (1 sp.), and Bacillus (2 spp.). The results show that douchi contains a large natural population of LAB of diverse composition from which some strains could be selected as starters for functional fermented foods. This is the first study on the original douchi from Yunnan, and the results suggest that it may be a useful source for the isolation of LAB. This study has also laid a foundation for further research on developing functional douchi products.

Members of group III histidine kinases from different filamentous fungi were previously shown to mediate osmoregulation and resistance to dicarboximide, phenylpyrrole and, aromatic hydrocarbon fungicides. In this study, we report the disruption of the gene encoding group III histidine kinase, AlHK1, in the economically important plant pathogen Alternaria longipes. The AlHK1 gene disruption had pleiotropic effects on this fungus. Besides the expected osmosensitivity and fungicides resistance, AlHK1 participated in the spore production process. In addition, the ΔAlHK1 strains had stronger aggressive ability to infect their host plant than that of their parental strain, the wild-type strain C-00, suggested that AlHK1 was involved in the pathogenicity of A. longipes and performed in this function by a negative manner. This is the first report to our knowledge.

In this paper, a rapid, simple and sensitive HPLC method to screen and accurately measure the level of phenyllactic acid (PLA) in lactic acid bacterial (LAB) MRS culture supernatant was established. Using this method, we successfully identified a Lactobacillus plantarum strainYM-5-2 which could highly produce PLA, the yield of PLA by L. plantarum YM-5-2 reached 147.91 mg/L. To our knowledge, this is the highest PLA-producing LAB strain when growing in MRS broth without any fermentation conditions optimizing, and it is the first report on antibacterial factor produced by LAB isolated from 'Douchi'-a traditional fermented soybean food in Yunnan Province, China. Antimicrobial activity of L. plantarum YM-5-2 suggested that it may be used as a biopreservation microbe in the food industry.

An extracellular 72-kDa xylanase from an Escherichia coli transformant that harbored pXED30 carrying xynE of Aeromonas caviae ME-1 was purified by extraction following SDS-PAGE to electrophoretic homogeneity. The analysis of N-terminal 10 amino acid residues (Gly-Ala-Arg-Ala-Gln-Ala-Ala-Ala-Asp-Val) revealed a 72-kDa product derived by the cleavage of Gly26-Gly27 at the N-terminal region of 110 kDa XynE. The 72-kDa xylanase from A. caviae ME-1 was found to have the same sequence as that of the N-terminal 10 amino acid residues. When OmpT-deficient E. coli mutants were used as hosts, the 72-kDa xylanase was detected in cell-free extracts, but not in the culture supernatant, suggesting that OmpT is not involved in the cleavage at the C-terminal region, but is involved in the secretion of 72-kDa xylanase to the culture medium.