PemK's Arg24 is a crucial residue for PemIK toxin-antitoxin system to induce the persistence of Weissella cibaria against ciprofloxacin stress.

The toxin-antitoxin (TA) system plays a key role in bacteria escaping antibiotic stress with persistence, however, the mechanisms by which persistence is controlled remain poorly understood. Weissella cibaria, a novel probiotic, can enters a persistent state upon encountering ciprofloxacin stress. Conversely, it resumes from the persistence when ciprofloxacin stress is relieved or removed. Here, it was found that PemIK TA system played a role in transitioning between these two states. And the PemIK was consisted of PemK, an endonuclease toxic to mRNA, and antitoxin PemI which neutralized its toxicity. The PemK specifically cleaved the U↓AUU in mRNA encoding enzymes involved in glycolysis, TCA cycle and respiratory chain pathways. This cleavage event subsequently disrupted the crucial cellular processes such as hydrogen transfer, electron transfer, NADH and FADH2 synthesis, ultimately leading to a decrease in ATP levels and an increase in membrane depolarization and persister frequency. Notably, Arg24 was a critical active residue for PemK, its mutation significantly reduced the mRNA cleavage activity and the adverse effects on metabolism. These insights provided a clue to comprehensively understand the mechanism by which PemIK induced the persistence of W. cibaria to escape ciprofloxacin stress, thereby highlighting another novel aspect PemIK respond for antibiotic stress.

Quantitative proteomic analysis reveals the mechanism and key esterase of ß-cypermethrin degradation in a bacterial strain from fermented food.

Beta-cypermethrin (ß-CY) residues in food are an important threat to human health. Microorganisms can degrade ß-CY residues during fermentation of fruits and vegetables, while the mechanism is not clear. In this study, a comprehensively investigate of the degradation mechanism of ß-CY in a food microorganism was conducted based on proteomics analysis. The ß-CY degradation bacteria Gordonia alkanivorans GH-1 was derived from fermented Pixian Doubanjiang. Its crude enzyme extract could degrade 77.11% of ß-CY at a concentration of 45 mg/L within 24 h. Proteomics analysis revealed that the ester bond of ß-CY is broken under the action of esterase to produce 3-phenoxy benzoic acid, which was further degraded by oxidoreductase and aromatic degrading enzyme. The up-regulation expression of oxidoreductase and esterase was confirmed by transcriptome and quantitative reverse transcription PCR. Meanwhile, the expression of esterase Est280 in Escherichia coli BL21 (DE3) resulted in a 48.43% enhancement in the degradation efficiency of ß-CY, which confirmed that this enzyme was the key enzyme in the process of ß-CY degradation. This study reveals the degradation mechanism of ß-CY by microorganisms during food fermentation, providing a theoretical basis for the application of food microorganisms in ß-CY residues.

Mechanism of deltamethrin biodegradation by Brevibacillus parabrevis BCP-09 with proteomic methods.

Ester-containing deltamethrin pesticides are widely used in farmland and have inevitable side effects on the biosphere and human health. Microbia have been used for efficient degradation of deltamethrin, but the related mechanism and enzyme characteristics have not been elucidated. In this study, a species Brevibacillus parabrevis BCP-09 could degrade up to 75 mg L-1 deltamethrin with a degradation efficiency of 95.41%. Proteomic and genomic methods were used to explore its degradation mechanism. Enzymes belonged to hydrolases, oxidases and aromatic compound degrading enzymes were expressed enhanced and might participate in the deltamethrin degradtion. RT-PCR experiment and enzyme activity analysis verified the degradation of deltamethrin by bacterial protein. Additionally, the formation of endospores can help strain BCP-09 resist the toxicity of deltamethrin and enhance its degradation. This study supplies a scientific evidence for the application of Brevibacillus parabrevis BCP-09 in the bioremediation of environmental pollution and enriches the resources of deltamethrin-biodegradable proteins.

Effect of apple polyphenols on physicochemical properties of pea starch/pulp cellulose nanofiber composite biodegradable films.

A pea starch (PS) and pulp cellulose nanofibers (CNF-P) hybrid matrix biodegradable film was prepared using apple polyphenol (AP) as the active substance. SEM and thermogravimetric analyses showed that apple polyphenols could be uniformly distributed and form hydrogen bonds with the matrix, and the increase in crystallinity improved the thermal stability of the films (the final residue of the films increased from 22.66 % to 31.82 %). The TS and EAB of the films reached their maximum values of 11.14 ± 1.73 MPa and 71.55 ± 8.8 %, respectively, at an AP content of 1.5 %. It should be noted that the antioxidant properties of the films were significantly positively correlated with the AP content, and the DPPH radical scavenging rate of the films reached 73.77 % at an AP content of 4.5 %, which was about 49 times higher than that of the control film. The same trend was observed in the UV-vis spectra. In addition, the total color difference and water solubility of the membranes increased from 4.29 ± 0.29 to 31.86 ± 1.90 and from 20.01 ± 0.97 % to 21.70 ± 1.99 %, respectively, and the biodegradability also showed an upward trend. These findings provide a theoretical basis and data support for the development of multifunctional biodegradable food packaging materials.

Inhibition of Perilla frutescens Essential Oil on Pellicle Formation of Candida tropicalis and Pichia kluyveri and Its Effect on Volatile Compounds in Sichuan Pickles.

Pellicle formation is the most typical characteristic of deteriorating fermented vegetable products. Perilla frutescens essential oil (PEO) is widely used as a useful natural preservative. However, few studies have addressed the antifungal activity and mechanism of PEO in pellicle formation microorganisms, and it is still unclear whether it can inhibit pellicle formation and affect its volatile compounds in Sichuan pickles. The current study showed that PEO can inhibit pellicle formation during fermentation of Sichuan pickles as it had significant antifungal activity against the pellicle formation microorganisms Candida tropicalis SH1 and Pichia kluyveri SH2. The minimum inhibitory concentration (MIC) of PEO against C. tropicalis SH1 and P. kluyveri SH2 was determined to be 0.4 µL/mL, and the minimum fungicidal concentrations (MFCs) were 1.6 µL/mL and 0.8 µL/mL, respectively. The antifungal mechanism was activated as a result of damage to the cell membrane, an increase in the cell permeability, a decrease in the mitochondrial membrane potential, and the inhibition of ATPase activity. Meanwhile, the addition of PEO to Sichuan pickles can enrich the profiles of volatile compounds during fermentation, including limonene, myrcene, 1,8-cineole, linalool, perilla ketone, heptanal, hexanal, α-thujone and ß-terpineol and thus improve the overall sensory acceptability. These results indicated that PEO has the potential to be used as a novel food preservative to control pellicle formation in fermented vegetables.

Whole genome sequencing and transcriptomics-based characterization of a novel ß-cypermethrin-degrading Gordonia alkanivorans GH-1 isolated from fermented foods.

Beta-cypermethrin (ß-CY) is an organic compound that is widely used as a synthetic pesticide in agriculture and family. Excessive accumulation of ß-CY inevitably causes environmental pollution, which has led to food safety and human health concerns. Identification of microorganisms from food sources that are capable of ß-CY biodegradation may help prevent pollution due to ß-CY accumulation. Here, Gordonia alkanivorans GH-1, which was isolated from the traditional Sichuan fermented food, Pixian Doubanjiang, could not only degrade 82.76% of 50 mg/L ß-CY at 96 h, but also degraded the intermediate degradation products including dibutyl phthalate (DBP), benzoic acid (BA) and phenol (Ph). This bacterial strain, thus, effectively improved the efficiency of removal of ß-CY and its related metabolites, without being limited by toxic intermediates. Whole genome sequencing and transcriptomics analyses have demonstrated that the bacteria affected the transcription of genes related to cell response and material transport under the stress induced by ß-CY, and thereby promoted degradation and transformation of ß-CY. Moreover, a complete pathway of ß-CY degradation is proposed based on the key genes involved in degradation. This study provides important theoretical significance and reference value for eliminating pesticide residues in agricultural products and food to ensure food safety.

Fungal Biomarkers in Traditional Starter Determine the Chemical Characteristics of Turbid Rice Wine from the Rim of the Sichuan Basin, China.

The fungal community in Qu plays a key role in the formation of turbid rice wine (TRW) style. The Sichuan Basin and its surrounding areas have become one of the main TRW production regions in China; however, the fungal community in Qu and how they affect the characteristics of TRW remain unknown. Therefore, this study provided insight into the fungal biomarkers in Qu from Guang'an (GQ), Dazhou (DQ), Aba (AQ), and Liangshan (LQ), as well as their relationships with compounds in TRW. The main biomarkers in GQ were Rhizopus arrhizus, Candida glabrata, Rhizomucor pusillus, Thermomyces lanuginosus and Wallemia sebi. However, they changed to Saccharomycopsis fibuligera and Mucor indicus in DQ, Lichtheimia ramose in AQ, and Rhizopus microsporus and Saccharomyces cerevisiae in LQ. As a response to fungal biomarkers, the reducing sugar, ethanol, organic acids, and volatile compounds were also changed markedly in TRWs. Among important volatile compounds (VIP > 1.00), phenethyl alcohol (14.1-29.4%) was dominant in TRWs. Meanwhile, 3-methyl-1-butanol (20.6-56.5%) was dominant in all TRWs except that fermented by GQ (GW). Acetic acid (29.4%) and ethyl palmitate (10.1%) were dominant in GW and LW, respectively. Moreover, GQ biomarkers were positively correlated with acetic acid and all unique important volatile compounds in GW. DQ biomarkers had positive correlations with unique compounds of acetoin and ethyl 5-chloro-1,3,4-thiadiazole-2-carboxylate in DW. Meanwhile, the AQ biomarkers were positively correlated with all AW unique, important, and volatile compounds. Although there were not any unique volatile compounds in LW, 16 important volatile compounds in LW were positively related to LQ biomarkers. Obviously, biomarkers in different geographic Qu played vital roles in the formation of important volatile compounds, which could contribute specific flavor to TRWs. This study provided a scientific understanding for future efforts to promote the excellent characteristics of TRW by regulating beneficial fungal communities.

Characterization and Bioactive Potential of Carotenoid Lutein from Gordonia rubripertncta GH-1 Isolated from Traditional Pixian Douban.

The characterization and bioactive properties of carotenoid produced by Gordonia rubripertincta GH-1 originating from Pixian Douban (PXDB), the Chinese traditional condiment, was investigated. The produced and purified yellow pigment was characterized by ultraviolet-visible spectroscopy (UV-Vis), Fourier transformed infrared (FTIR), nuclear magnetic resonance (NMR), and high-resolution mass spectrometry (HRMS), and was identified as carotenoid lutein. Additionally, the bioactive activity of lutein from G. rubripertincta GH-1 was evaluated by measuring the free radical scavenging capacity in vitro and feeding zebrafish lutein through aqueous solution. The results showed that the carotenoid lutein had strong antioxidant capacity and a protective effect on zebrafish eye cells, which could inhibit the apoptosis of eye cells in a concentration dependent manner. The results suggested that carotenoid lutein from G. rubripertincta GH-1 could be utilized as a potential source of natural antioxidants or functional additives for food/pharmaceutical industries.

Isolation and characterization of bacteria that produce quorum sensing molecules during the fermentation and deterioration of pickles.

Pickles are typical traditional Chinese fermented vegetables. Complex microbiota interacts throughout the fermentation and deterioration process. Minimal studies are available involving quorum sensing (QS) signaling in pickles. This study investigated the changes in the general pickle properties and microbial diversity at 4 d, 31 d, and 79 d. The QS signaling activity of various strains isolated at these key time points was screened using biosensor strains, while the types of signal molecules were further identified using UHPLC/QTOF-MS/MS. At 4 d, Lactobacillus represented the dominant genus, while Lactobacillus plantarum was identified as the bacteria with AI-2-producing ability. At 31 d, the dominant genus was also Lactobacillus, while the relative abundance of Pediococcus displayed a distinct increase. At this time point, L. plantarum represented the AI-2-producing bacteria, followed by Lactobacillus brevis, Pediococcus sp., Enterobacter sp., and Bacillus megaterium. At 79 d, Lactobacillus was displaced by Enterobacter as the dominant microorganisms, while the AI-2-producing bacteria were identified as L. plantarum, Enterobacter sp., B. megaterium, Klebsiella sp., and Staphylococcus sp. Moreover, AHL activity was only present in isolates from the 79-d brine and was identified as C4-HSL and C6-HSL. In addition, the luxS gene was amplified via cDNA reversely transcription from the total RNA extracted from the brine at all three time points using the L. plantarum luxS primers. The AHL-related genes were only amplified in the RNA of 79-d brine samples using Klebsiella pneumoniae- and Bacillus cereus-related primers. This study presented theoretical references for QS during pickle fermentation and deterioration.

HigBA toxin-antitoxin system of Weissella cibaria is involved in response to the bile salt stress.

BACKGROUND: Toxin-antitoxin (TA) systems are prevalent adaptive genetic elements in bacterial genomes, which can respond to environmental stress. While, few studies have addressed TA systems in probiotics and their roles in the adaptation to gastrointestinal transit (GIT) environments. RESULTS: The Weissella cibaria 018 could survive in pH 3.0-5.0 and 0.5-3.0 g L-1 bile salt, and its HigBA system responded to the bile salt stress, but not to acid stress. The toxin protein HigB and its cognate antitoxin protein HigA had 85.1% and 100% similarity with those of Lactobacillus plantarum, respectively, and they formed the stable tetramer HigB-(HigA)2 -HigB structure in W. cibaria 018. When exposed to 1.5-3.0 g L-1 bile salt, the transcriptions of higB and higA were up-regulated with 4.39-19.29 and 5.94-30.91 folds, respectively. Meanwhile, W. cibaria 018 gathered into a mass with 48.07% survival rate and its persister cells were found to increase 8.21% under 3.0 g L-1 bile salt. CONCLUSION: The HigBA TA system of W. cibaria 018 responded to the bile salt stress, but not to acid stress, which might offer novel perspectives to understand the tolerant mechanism of probiotics to GIT environment. © 2022 Society of Chemical Industry.

The Use of γ-Aminobutyric Acid-Producing Saccharomyces cerevisiae SC125 for Functional Fermented Beverage Production from Apple Juice.

The development of functional fermented beverages enriched with γ-aminobutyric acid (GABA) has been pursued because of the health benefits of GABA; however, few studies have described GABA production by yeast. Therefore, this study aimed to produce fermented apple beverages enriched with GABA produced by Saccharomyces cerevisiae SC125. Golden Delicious apples were fermented by S. cerevisiae SC125 to produce a novel functional beverage; commercial yeast was used as the control. The GABA, organic acid, and volatile compound content during the fermentation process was investigated by high-performance liquid chromatography and headspace solid-phase microextraction/gas chromatography-mass spectrometry. A yield of 898.35 ± 10.10 mg/L GABA was achieved by the efficient bioconversion of L-monosodium glutamate. Notably, the S. cerevisiae SC125-fermented beverage produced several unique volatile compounds, such as esters, alcohols, 6-decenoic acid, and 3-hydroxy-2-butanone, and showed significantly enhanced contents of organic acids, including malic acids, citric acid, and quinic acid. Sensory analysis demonstrated that the S. cerevisiae SC125-fermented apple beverage had improved aroma, flavor, and overall acceptability. In conclusion, a fermented functional apple beverage containing GABA was efficiently produced using S. cerevisiae SC125.

Tetracycline residue alters profile of lactic acid bacterial communities and metabolites of ginger pickle during spontaneous fermentation.

Antibiotic residue, an emerging contaminant, has been an increasing concern worldwide in vegetables. However, the focus is still limited to its accumulation in vegetables and its adverse health effect on humans, with little mention of its impact on the vegetable process. Fermentation is an important method of the vegetable process. Tetracycline (TC) is the most widely used antibiotic, and its residue is often higher than other antibiotics in ginger. Therefore, current research was to characterize how the TC residue affected the lacticacidbacterial (LAB) communities and metabolites during the salted fermentation of ginger. The TC residue, organic acids, volatile compounds and LABs were analyzed during the fermentation, and the correlations between the LABs and compounds were also revealed. The results suggested that the hetero-lactic fermentation did not happen under the TC residue although the TC residue decreased from 4 mg/kg to 2.56 mg/kg in salt brine of ginger fermentation. Meanwhile, TC residue affected the occurrence, propagation and succession of LABs. The LAB biomarkers shifted to Lab. parafarraginis, Lab. buchneri and Lab. kisonensis under TC residue. Responded to LAB communities, the organic acids and volatile compounds were also markedly changed under the TC residue. And the important volatile compound variables shifted to citronellol, allylsenevol, geranyl acetate, vinylstearylether, isothiocyanic acid phenethyl, 3-octanol, geraniol, bingpian, citral and camphor. The transformation of LAB biomarkers induced by TC residue was the main cause of the change of important volatile compound variables, but interestingly, not all of them had significant positive or negative correlations. These results indicated that the antibiotic residue has an adverse ecological effect on the vegetable fermentation process.Therefore, the antibiotic residue should be listed as a quality control index in fermented vegetable raw materials.

Characterization and Antioxidant Activity of Released Exopolysaccharide from Potential Probiotic Leuconostoc mesenteroides LM187.

A released exopolysaccharide (rEPS)-producing strain (LM187) with good acid resistance, bile salt resistance, and cholesterol-lowering properties was isolated from Sichuan paocai and identified as Leuconostoc mesenteroides subsp. mesenteroides. The purified rEPS, designated as rEPS414, had a uniform molecular weight of 7.757 × 105 Da. Analysis of the monosaccharide composition revealed that the molecule was mainly composed of glucose. The Fourier transform-infrared spectrum showed that rEPS414 contained both α-type and ß-type glycosidic bonds. 1H and 13C nuclear magnetic resonance spectra analysis showed that the purified rEPS contained arabinose, galactose, and rhamnose, but less uronic acid. Scanning electron microscopy demonstrated that the exopolysaccharide displayed a large number of scattered, fluffy, porous cellular network flake structures. In addition, rEPS414 exhibited strong in vitro antioxidant activity. These results showed that strain LM187 and its rEPS are promising probiotics with broad prospects in industry.

Metabolic profiles of Lactobacillus paraplantarum in biofilm and planktonic states and investigation of its intestinal modulation and immunoregulation in dogs.

The use of probiotics has recently become a considerably promising research area. The most advanced fourth-generation probiotics involve beneficial bacteria enclosed in biofilms. However, differences in the effects of probiotics in biofilm and those in planktonic states are, as yet, unclear. In this study, it was ascertained that the biofilm mode of Lactobacillus paraplantarum L-ZS9 had a comparatively higher density and stronger resistance. Untargeted metabolomics analysis suggested a significant distinction between planktonic and biofilm cells, with amino acids and carbohydrate metabolism both more active in the biofilm mode. Furthermore, the in vivo experiment showed that the biofilm strain displayed better immunomodulation activity, which could increase the relative abundance of Lactobacillus in the intestinal microbiota of dogs. The relative abundance of intestinal microbiota participating in carbohydrate metabolism was higher in the biofilm probiotic-treated dogs. Correlation analysis between L-ZS9-producing metabolites, dog intestinal microbiome diversity and dog blood immune indexes (sIgA or IgG) revealed the interaction between these three components, which might explain the mechanisms by which biofilm L-ZS9 regulated the intestinal microbiome and immunity activity of the host, through the production of various metabolites. Findings of this study will, thus, enhance understanding of the beneficial effects of biofilm probiotics, as well as provide references for further investigation.

Microbiota Succession and Chemical Composition Involved in the Radish Fermentation Process in Different Containers.

Traditional Chinese fermented vegetables are a type of brine-salted fermented vegetable product. During the spontaneous fermentation, various compounds are produced, degraded, and converted, influencing the quality of the fermented pickle. To ascertain the effect of different containers on the fermentation process of the pickles, this study investigated the bacterial diversity and the chemical composition characteristics of the pickle (radish) fermented in commonly used containers including glass jars (GL), porcelain jars (PO), and plastic jars (PL). The correlation between chemical compounds and microbial community was further analyzed. The changes in pH values suggested that PL may facilitate the quickest fermentation of the pickles, while the process in PO progressed at the lowest rate. The PL brine samples contained higher levels of lactic acid and threonine, while more abundant volatile chemical compounds were evident in PO. The container materials had no significant influence on the microbial structure, wherein Lactobacillus was the absolute dominant genus in all containers. But container material did have an effect on the abundance of specific genus, such as Lactococcus and Pediococcus. The correlation between these major genera was also analyzed and gene function prediction indicated that the top three pathways were: carbohydrate metabolism, amino acid metabolism, and energy metabolism. Lactobacillus negatively correlated with methionine, tyrosine, lysine, and arginine, but positively correlated with ammonia, and lactic acid and acetic acid both just correlated with Pediococcus. This study provides new insights into the microbiota succession and chemical compounds involved in the vegetable fermentation.

Characterization of γ-aminobutyric acid (GABA)-producing Saccharomyces cerevisiae and coculture with Lactobacillus plantarum for mulberry beverage brewing.

One elite γ-aminobutyric acid (GABA) producing strain of Saccharomyces cerevisiae SC125 was isolated and identified from Chinese traditional paocai. S. cerevisiae SC125 and Lactobacillus plantarum BC114 were used as cooperative species to ferment mulberry (Morus alba L.) and produce a novel beverage enriched with GABA. The GABA, organic acids and volatile compounds in different fermentation stages were determined by high-performance liquid chromatography (HPLC) and headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). It was noted that coculture changed the profiles of flavor compounds in mulberry beverage. The tartaric and succinic acid contents increased to 1.34 g/L and 0.39 g/L, respectively. Lactic, malic, citric, and oxalic acid levels ranged between 0.92 and 2.56 g/L, and ethanol and glycerol were produced at 2.66 g/L and 1.81 g/L, respectively. More volatile compounds were detected in the coculture with significantly enhanced concentrations of fruity esters including ethyl caproate, ethyl propionate, butyl acetate, ethyl butyrate, ethyl caprylate and ethyl caprate, and alcohols of phenylethyl alcohol, 1-pentanol and 2-amino-1,3-propanediol. Also, a yield of 2.42 g/L GABA was achieved in the coculture. In conclusion, S. cerevisiae SC125 and L. plantarum BC114 coculture promotes the production of flavor compounds and GABA in mulberry beverage brewing.

Mucor indicus and Rhizopus oryzae co-culture to improve the flavor of Chinese turbid rice wine.

BACKGROUND: One of the most important species used to ferment Chinese turbid rice wine (CTRW) at an industrial-scale level is Rhizopus oryzae, although the flavor of CTRW fermented by pure R. oryzae is inferior to that of traditional CTRW. RESULTS: Mucor indicus was used as a cooperative species to improve the flavor of CTRW presented by R. oryzae. The flavor compounds in different fermentation stages were determined by headspace solid-phase microextraction-gas chromatography-mass spectrometry and high-performance liquid chromatography. It was noted that the M. indicus and R. oryzae co-culture changed the profiles of flavor compounds in CTRW, including esters, higher alcohols, amino acids and organic acids, and also significantly enhanced the concentration of sweet amino acids, fruity and floral esters, and higher alcohols. Sensory evaluation demonstrated that the CTRW fermented by M. indicus and R. oryzae had a more intense aroma, harmonious taste, continuation and full body mouth-feel because of more abundant flavor compounds. CONCLUSION: Mucor indicus is a promising species for co-culture with R. oryzae to improve the flavor of CTRW. © 2019 Society of Chemical Industry.

Organic Houttuynia cordata Thunb harbors higher abundance and diversity of antibiotic resistance genes than non-organic origin, suggesting a potential food safe risk.

The organic agricultural products has been growing rapidly in recent years. However, a potential food safe risk, resulted by introduction more antibiotic resistant genes (ARGs) accompanied with animal manure using to organic farming, has long been overlooked. In current study, the bacterial community, 22 tetracycline, 3 aminoglycoside and 4 ß-lactams ARGs were respectively investigated in the organic, chemical and wild Houttuynia cordata Thunb (HCT). A total of 9 tetracycline, 3 aminoglycoside and 2 ß-lactam ARG subtypes were detected, and the organic HCT harbored more ARG subtypes. The absolute and relative abundance of total ARGs in organic HCT was strikingly higher than that in chemical and wild HCT. The Enterobacteriaceae, Aeromonadaceae, Pseudomonadceae, Moraxellaceae and Oxalobacteraceae were the dominant taxa in the chemical and wild HCT, but in the organic HCT, only Enterobacteriaceae posed 83.23% - 87.40% of bacterial community. Fourteen bacterial families might be the possible hosts of ARG subtypes in the HCT. Enterobacteriaceae was a possible host of most ARG subtypes, including tetA, tetB, tetC, tetE and aadA, and it was the main bacteria affecting the behavior of ARGs in the HCT. Additionally, the tetracycline ARG subtypes had more possible hosts. These results help to better understand the ARG potential food safe risk and develop effective measures to prevent the ARG dissemination in organic agricultural product.

Microbial succession in the traditional Chinese Luzhou-flavor liquor fermentation process as evaluated by SSU rRNA profiles.

The community succession of microbes inhabited in the fermenting lees of Luzhou-flavor liquor was investigated based on small-subunit rRNA culture independent method. All sequences recovered from fermenting lees respectively fell into the genera of Lactobacillus, Streptococcus, Bacillus, Staphylococcus, Clostridium, Pelobacter, Actobacter, Serratia, Burkholderia, Rhodoccous, Corynebacterium, Arthrobacter, Microbacterium, Curtobacterium, Leptotrichia, Methanocuuleus, Saccharomyces, Zygosaccharomyces, Saccharomycopsis, Pichia, Talaromyces, Aspergillus, Eurotium, Fomitopsis and Trichosporon. The fungal Pichia, Saccharomycopsis and Talaromyces were most abundant in the lees fermented for 1 day, the fungal Eurotium and the bacteria Burkholderia, Streptococcus and Lactobacillus were dominant in the lees fermented for 7 days, only the bacteria Lactobacillus, Burkholderia were prevalent in the lees fermented for 60 days. Most genera almost existed in the fermenting lees, while their distributions were significantly different in 1, 7 and 60 days fermented lees. The prokaryotic community similarity coefficient was from 0.5000 to 0.5455 and followed to 0.1523, and that of eukaryotic community was from 0.5466 to 0.5259 and to 0.3750 when compared at species level. These results suggested that many microbes in lees have community successions associated with fermenting and that such successions maybe contribute the fermentation process of Luzhou-flavor liquor and is main reasons that the characteristic flavor factors are produced.

Microbial Diversity and Succession in the Chinese Luzhou-Flavor Liquor Fermenting Cover Lees as Evaluated by SSU rRNA Profiles.

The microbial diversity and the community succession in the fermenting cover lees of Chinese Luzhou-flavor liquor were investigated by small-subunit rRNA (SSU rRNA) culture independent method. All sequences retrieved from the 1, 7 and 60 days fermented cover lees were respectively assigned into the genera of Streptococcus, Acetobacter, Arthrobacter, Bacillus, Staphylococcus, Serratia, Nocardia, Methanoculleus, Clostridium, Aneurinibacillus, Corynebacterium, Lactobacillus, Microbacterium, Trichosporon, Saccharomycopsis, Sagenomella, Talaromyces, Eurotium, Issatchenkia, Zygosaccharomyces, Saccharomyces and TM7 phylum. The fungal Issatchenkia, Saccharomycopsis and Talaromyces and the bacteria Staphylococcus and Lactobacillus were most abundant in the 1 day fermented cover lees, the fungal Issatchenkia, Saccharomyces and Talaromyces and the bacteria Bacillus and Streptococcus were dominant in the 7 days cover lees, the archaea Methanoculleus and the fungal Eurotium and Talaromyces were prevalent in the 60 days cover lees. When the microbial community profiles in three samples were compared at species level, the prokaryotic community similarity coefficient was from 0.4042 to 0.5703 and descended to 0.2222, and that of eukaryotic community was from 0.3000 to 0.6000 and followed to 0.5215. These results suggested that microbial diversity variability and community succession have happened in the cover lees associated with fermentation proceeding and such variability and succession respond for the appearance of some unique flavor of Luzhou-flavor liquor.