Recent advances in developing butyrogenic functional foods to promote gut health.

As one of the major short-chain fatty acids produced via microbial fermentation, butyrate serves as not only a preferred energy substrate but also an important signaling molecule. Butyrate concentrations in circulation, tissues, and gut luminal contents have important pathophysiological implications. The genetic capacity of butyrate biosynthesis by the gut microbiota is frequently compromised during aging and various disorders, such as inflammatory bowel disease, metabolic disorders and colorectal cancer. Substantial efforts have been made to identify potent butyrogenic substrates and butyrate-hyperproducing bacteria to compensate for butyrate deficiency. Interindividual butyrogenic responses exist, which are more strongly predicted by heterogeneity in the gut microbiota composition than by ingested prebiotic substrates. In this review, we catalog major food types rich in butyrogenic substrates. We also discuss the potential of butyrogenic foods with proven properties for promoting gut health and disease management using findings from clinical trials. Potential limitations and constraints in the current research are highlighted. We advocate a precise nutrition approach in designing future clinical trials by prescreening individuals for key gut microbial signatures when recruiting study volunteers. The information provided in this review will be conducive to the development of microbiota engineering approaches for enhancing the sustained production of butyrate.

Screening of Lactobacillus plantarum Subsp. plantarum with Potential Probiotic Activities for Inhibiting ETEC K88 in Weaned Piglets.

For screening excellent lactic acid bacteria (LAB) strains to inhibit enterotoxigenic Escherichia coli (ETEC) K88, inhibitory activities of more than 1100 LAB strains isolated from different materials, and kept in the lab, were evaluated in this study. Nine strains with inhibition zones, at least 22.00 mm (including that of a hole puncher, 10.00 mm), and good physiological and biochemical characteristics identified by 16S DNA gene sequencing and recA gene multiple detection, were assigned to Lactobacillus (L.) plantarum subsp. plantarum (5), L. fermentum (1), L. reuteri (1), Weissella cibaria (1) and Enterococcus faecalis (1), respectively. As investigated for their tolerance abilities and safety, only strain ZA3 possessed high hydrophobicity and auto-aggregation abilities, had high survival rate in low pH, bile salt environment, and gastrointestinal (GI) fluids, was sensitive to ampicillin, and resistant to norfloxacin and amikacin, without hemolytic activity, and did not carry antibiotic resistance genes, but exhibited broad spectrum activity against a wide range of microorganisms. Antibacterial substance may attribute to organic acids, especially lactic acid and acetic acid. The results indicated that the selected strain L. plantarum subsp. plantarum ZA3 could be considered a potential probiotic to inhibit ETEC K88 in weaned piglets for further research.

Screening of Probiotic Activities of Lactobacilli Strains Isolated from Traditional Tibetan Qula, A Raw Yak Milk Cheese.

In this study, 69 lactobacilli isolated from Tibetan Qula, a raw yak milk cheese, were screened for their potential use as probiotics. The isolates were tested in terms of: Their ability to survive at pH 2.0, pH 3.0, and in the presence of 0.3% bile salts; tolerance of simulated gastric and intestinal juices; antimicrobial activity; sensitivity against 11 specific antibiotics; and their cell surface hydrophobicity. The results show that out of the 69 strains, 29 strains (42%) had survival rates above 90% after 2 h of incubation at pH values of 2.0 or 3.0. Of these 29 strains, 21 strains showed a tolerance for 0.3% bile salt. Incubation of these 21 isolates in simulated gastrointestinal fluid for 3 h revealed survival rates above 90%; the survival rate for 20 of these isolates remained above 90% after 4 h of incubation in simulated intestinal fluid. The viable counts of bacteria after incubation in simulated gastric fluid for 3 h and simulated intestinal fluid for 4 h were both significantly different compared with the counts at 0 h (p<0.001). Further screening performed on the above 20 isolates indicated that all 20 lactobacilli strains exhibited inhibitory activity against Micrococcus luteus ATCC 4698, Bacillus subtilis ATCC 6633, Listeria monocytogenes ATCC 19115, and Salmonella enterica ATCC 43971. Moreover, all of the strains were resistant to vancomycin and streptomycin. Of the 20 strains, three were resistant to all 11 elected antibiotics (ciprofloxacin, erythromycin, tetracycline, penicillin G, ampicillin, streptomycin, polymyxin B, vancomycin, chloramphenicol, rifampicin, and gentamicin) in this study, and five were sensitive to more than half of the antibiotics. Additionally, the cell surface hydrophobicity of seven of the 20 lactobacilli strains was above 70%, including strains Lactobacillus casei 1,133 (92%), Lactobacillus plantarum 1086-1 (82%), Lactobacillus casei 1089 (81%), Lactobacillus casei 1138 (79%), Lactobacillus buchneri 1059 (78%), Lactobacillus plantarum1141 (75%), and Lactobacillus plantarum 1197 (71%). Together, these results suggest that these seven strains are good probiotic candidates, and that tolerance against bile acid, simulated gastric and intestinal juices, antimicrobial activity, antibiotic resistance, and cell surface hydrophobicity could be adopted for preliminary screening of potentially probiotic lactobacilli.

Effects of Probiotic-Fermented Feed on the Growth Profile, Immune Functions, and Intestinal Microbiota of Bamei Piglets.

Purebred Bamei piglets present problems, including slow growth, respiratory disease, and post-weaning stress. This study investigated the effects of Lactobacillus plantarum QP28-1- and Bacillus subtilis QB8-fermented feed supplementation on the growth performance, immunity, and intestinal microflora of Bamei piglets from Qinghai, China. A total of 48 purebred Bamei piglets (25 days; 6.8 ± 0.97 kg) were divided into the following four groups for a 28-day diet experiment: basal feed (CK); diet containing 10% Lactobacillus plantarum-fermented feed (L); diet containing 10% Bacillus subtilis-fermented feed (B); and diet containing a mixture of 5% Lactobacillus plantarum + 5% Bacillus subtilis-fermented feed (H). The daily weight gain and daily food intake of group H increased (p < 0.05), and the feed/weight gain ratios of the groups fed with fermented feed decreased more than that of the CK group. The levels of three immune factors, namely immunoglobulin (Ig)M, IgG, and interferon-γ, were higher (p < 0.05), whereas those of tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6 were lower (p < 0.05) in the fermented feed groups than in the CK group. Total protein was higher (p < 0.05), while urea nitrogen, total cholesterol and triglycerides were lower (p < 0.05) in the mixed-fermented feed group than in the CK group. Analysis of the gut microbiota showed that the addition of fermented feed increased the α-diversity of the gut microbiota, increasing the abundances of probiotics including Lactobacillus, Muribaculaceae, Ruminococcaceae, Prevotellaceae, and Rikenellaceae. Additionally, correlation analysis demonstrated that several of these probiotic bacteria were closely related to serum immunity. In conclusion, fermented feed supplementation rebuilt the intestinal microbiota of Bamei piglets, thereby reducing the feed/weight ratio, improving feed intake, and enhancing immunity.

Investigation on Fermentation Characteristics and Microbial Communities of Wheat Straw Silage with Different Proportion Artemisia argyi.

Mycotoxins, secondary metabolites of fungi, are a major obstacle to the utilization of animal feed for various reasons. Wheat straw (WS) is hollow, and miscellaneous bacteria can easy attach to its surface; the secondary fermentation frequency after silage is high, and there is a risk of mycotoxin poisoning. In this study, a storage fermentation process was used to preserve and enhance fermentation quality in WS through the addition of Artemisia argyi (AA), which is an effective method to use WS resources and enhance aerobic stability. The storage fermentation of WS treated with AA had lower pH and mycotoxin (AFB1 and DON) values than the control due to rapid changes in microbial counts, especially in the 60% AA groups. Meanwhile, the addition of 60% AA improved anaerobic fermentation profiles, showing higher lactic acid contents, leading to increased efficiency of lactic acid fermentation. A background microbial dynamic study indicated that the addition of 60% AA improved the fermentation and aerobic exposure processes, decreased microbial richness, enriched Lactobacillus abundance, and reduced Enterobacter and Aspergillus abundances. In conclusion, 60% AA treatment could improve the quality by increase fermentation quality and improve the aerobic stability of WS silage by enhancing the dominance of desirable Lactobacillus, inhibiting the growth of undesirable microorganisms, especially fungi, and reducing the content of mycotoxins.

Effects of Lactic Acid Bacteria Reducing the Content of Harmful Fungi and Mycotoxins on the Quality of Mixed Fermented Feed.

The contamination of fermented feeds and foods with fungi and mycotoxins is a major food safety issue worldwide. Certain lactic acid bacteria (LAB), generally recognized as safe (GRAS) fermentation probiotics, are able to reduce microbial and mycotoxins contamination. In this study, Lactiplantibacillus (L.) plantarum Q1-2 and L. salivarius Q27-2 with antifungal properties were screened as inoculants for mixed fermenting feed, and the fermentation and nutritional qualities, microbial community, and mycotoxins of mixed fermented feed were analyzed at different fermentation periods (1, 3, 7, 15, and 30 days, respectively). The findings indicated that the utilization of Q1-2 and Q27-2 strains in fermenting feed led to a decrease in pH and an increase in lactic acid concentration and the proportion of Lactiplantibacillus, while effectively restraining the proliferation of undesirable microorganisms. In particular, Q1-2 reduced the relative abundance of fungi including Fusarium and Aspergillus. Compared to the control group, the Q1-2 and Q27-2 groups reduced aflatoxin B1 by 34.17% and 16.57%, and deoxynivalenol by up to 90.61% and 51.03%. In short, these two LAB inoculants could reduce the contents of aflatoxin B1 and deoxynivalenol to the limited content levels stipulated by the Chinese National Standard GB 13078-2017. These findings suggest that the LAB strains of Q1-2 and Q27-2 have potential applications in the feed industry for the mitigation of mycotoxin pollution, thereby enhancing the quality of animal feed.

Reclassification of Lactobacillus kimchii and Lactobacillus bobalius as later subjective synonyms of Lactobacillus paralimentarius.

Characterization and identification of strain CW 1 ( = JCM 17161) isolated from corn silage were performed. Strain CW 1 was a Gram-positive, catalase-negative and homofermentative rod that produced the DL-form of lactic acid. This strain exhibited more than 99.6% 16S rRNA gene sequence similarity and greater than 82% DNA-DNA reassociation with type strains of Lactobacillus kimchii, L. bobalius and L. paralimentarius. To clarify the taxonomic positions of these type strains, phenotypic characterization, 16S rRNA gene sequencing, ribotyping and DNA-DNA relatedness were examined. The three type strains displayed different L-arabinose, lactose, melibiose, melezitose, raffinose and N-acetyl-ß-glucosaminidase fermentation patterns. Phylogenetic analysis showed that L. paralimentarius is a closer neighbour of L. kimchii and L. bobalius, sharing 99.5-99.9% 16S rRNA gene sequence similarity, which was confirmed by the high DNA-DNA relatedness (≥82%) between L. paralimentarius JCM 10415(T), L. bobalius JCM 16180(T) and L. kimchii JCM 10707(T). Therefore, it is proposed that L. kimchii and L. bobalius should be reclassified as later synonyms of L. paralimentarius.

Characterization on structure and bioactivities of an exopolysaccharide from Lactobacillus curvatus SJTUF 62116.

This study aimed to investigate the chemical structure, physicochemical properties, antioxidant capacity, antibacterial ability and anti-biofilm formation activity of an exopolysaccharide (EPS) produced by Lactobacillus curvatus SJTUF 62116 from the fish Gymnocypris przewalskii. The purified EPS, denoted as EPS-1, was mainly composed of glucose and mannose at a relative molar ratio of 1:1.05 with molecular weight of 31.9 kDa. The chemical structure of EPS-1 was consisted of →2)-α-D-Manp-(1→, →4)-α-D-Manp-(1→, →3,6)-α-D-Manp-(1→, T-ß-D-Glcp-(1→, →6)-ß-D-Glcp-(1→, and →3)-ß-D-Glcp-(1→ glycosidic bonds. A sheet-like structure of dried EPS-1 was determined by scanning electron microscope (SEM), whilst a peak-shaped structure of EPS-1 was observed by atomic force microscope (AFM). The degradation temperature of EPS-1 was determined as 300.21 °C using thermogravimetric analysis (TGA). Moreover, the antioxidant capacity of EPS-1 at a concentration of 5.0 mg/mL against DPPH and ABTS was 84.50% and 92.53%, respectively. Furthermore, EPS-1 exhibited acceptable bacteriostatic efficacy against S. Enteritidis, E. coli, and S.aureus with significant inhibition of S. Enteritidis biofilm formation.

Microbial Population Succession and Community Diversity and Its Correlation with Fermentation Quality in Soybean Meal Treated with Enterococcus faecalis during Fermentation and Aerobic Exposure.

This study assessed the effects of Enterococcus faecalis (E. faecalis) in combination with protease on fermentation characteristics and microbial communities during ensiling and aerobic exposure phases of soybean meal (SBM). In this study, response surface methodology (RSM) was used to optimize the optimal growth conditions of E. faecalis ZZUPF95, which produced protease, and fermented SBM under the optimal fermentation conditions. The fermentation test was divided into four groups as follows: CK (Control check), ZZUPF95, Protease and ZZUPF95+Protease groups. Results showed that the best medium ratio of ZZUPF95 was glucose 1%, peptone 2%, inorganic salt 1.47%; fermentation time 36 h, inoculation amount 10%, ratio of material to water 1:1 is the optimal fermentation scheme; after fermentation and aerobic exposure treatment, ZZUPF95 and ZZUPF95 + Protease group can reduce the pH of feed, improve the content of lactic acid in the fermentation system, and have the effect of inhibiting the reproduction of pathogenic bacteria, increasing the content of crude protein and ether extract, and degrading crude fiber; the microbial community of SBM were changed after fermentation and aerobic exposure. This study explored the changes of fermentation quality of SBM, which has certain theoretical value to improve the fermentation mode and storage of SBM.

Microbial community and fermentation characteristic of whole-crop wheat silage treated by lactic acid bacteria and Artemisia argyi during ensiling and aerobic exposure.

Whole-crop wheat silage (WCWS) is an excellent feed material for ruminants. However, microbial fermentation during silage production consumes valuable nutrients, decreasing the quality of silage. The main objective of this study was to assess how the addition of increasing amounts of Artemisia argyi (AA) affected fermentation quality, microbial composition, and mycotoxin production in whole-crop wheat at dough stage (WCWD) silage during ensiling to aerobic exposure compared with Lactiplantibacillus buchneri (LB). The addition of 20% AA, resulted in a lower pH and higher lactic acid content, was found in silage treated with 20% AA, and an obvious increase in the relative abundance of Lactobacillus was detected in silages treated with LB and 20% AA, respectively. Meanwhile, inoculation with 20% AA decreased the abundance of harmful microorganisms, including Acinetobacter, Enterobacter, and Aspergillus. It also reduced the contents of mycotoxins, Aflatoxin B1 (AFB1), and deoxynivalenol (DON) during ensiling and aerobic exposure. These results confirmed that WCWD treated with 20% AA could improve the fermentation quality and enhance the aerobic stability of silage.

The Effect of Dietary Lactic Acid Bacteria on Intestinal Microbiota and Immune Responses of Crucian Carp (Carassius auratus) Under Water Temperature Decrease.

Temperature changes have a great impact on fish feeding, intestinal microorganisms, metabolism, and immune function. Therefore, it is necessary to develop effective methods to enhance the survival rates and growth of fish under water temperature changes. Lactic acid bacteria (LAB) are promising immunostimulatory feed additive, as demonstrated by their beneficial effects in several fish species. This study investigated the short-term effects of dietary LAB on intestinal microbiota composition and immune responses of crucian carp (Carassius auratus) when water temperature decreased from 30 ± 1°C to 18 ± 1°C. Lactococcus (L.) lactis 1,209 and L. lactis 1,242 with potential probiotics isolated from the intestine of Qinghai naked carp (Gymnocypris przewalskii) were selected as feed additives for the crucian carp feeding experiment. A total of 225 commercially available healthy crucian carp (250 ± 10 g) of similar age were kept in 30°C water for a week and then immediately transferred to 18 ± 1°C water, assigned to three dietary treatments for a 16-day feeding trial randomly: (1) HC, diets without additives (the control group); (2) HT, diets with 106 CFU/ml L. lactis 1,209; and (3) HL, with 106 CFU/ml L. lactis 1,242. Each group was set up with 3 replicates and each with 25 fish. The results showed that the mortality rate of crucian carp in HC, HT, and HL group was 50, 27, and 33%, respectively. High-throughput sequencing results displayed that the composition of the intestinal microorganism varied dynamically in response to different treatments and water temperature decrease. Among them, compared with the HC group, a higher abundance of Firmicutes and Proteobacteria, and a lower of Actinobacteria appeared in HT and HL. The cytokines heat shock protein 70 (HSP-70) in crucian carp intestinal tract significantly decreased when water temperature decreased (p < 0.05).

Variation of Microbial Community and Fermentation Quality in Corn Silage Treated with Lactic Acid Bacteria and Artemisia argyi during Aerobic Exposure.

Silage, especially whole crop corn silage (WCCS), is an important part of ruminant diets, with its high moisture content and rich nutrient content, which can easily cause contamination by mold and their toxins, posing a great threat to ruminant production, food safety and human health. The objective of this study was to examine effects of lactic acid bacteria (LAB) Lactiplantibacillus (L.) plantarum subsp. plantarum ZA3 and Artemisia argyi (AA) on the fermentation characteristics, microbial community and mycotoxin of WCCS during 60 days (d) ensiling and subsequent 7 d aerobic exposure. The results showed that WCCS treated with LAB and AA both had lower pH value and ammonia nitrogen (NH3-N) contents, and higher lactic and acetic acids concentration compared with other groups after 60 d ensiling. In addition, for microbial communities, Acetobacter and Enterobacter were inhibited in all AA group, while higher abundance of Lactobacilli was maintained; besides, Candida, Pichia and Kazachstania abundances were decreased in both 6% and 12% AA groups. The content of five kinds of mycotoxins were all significantly lower after 7 d of aerobic exposure. As for the total flavonoid (TF), which is significantly higher in all AA treated groups, it was positively correlated with Paenibacillus, Weissella and Lactobacilli, and negatively with Acetobacter, Enterobacteria, Kazachstania and Pichia.

Bacteriocin-Producing Lactic Acid Bacteria Strains with Antimicrobial Activity Screened from Bamei Pig Feces.

Lactic acid bacteria (LAB), which are characterized by producing various functional metabolites, including antioxidants, organic acids, and antimicrobial compounds, are widely used in the food industry to improve gut health and prevent the growth of spoilage microorganisms. With the continual incidence of foodborne disease and advocacy of consumers for gut health, LAB have been designated as vital biopreservative agents in recent years. Therefore, LAB with excellent antimicrobial properties and environmental tolerance should be explored further. In this study, we focus on screening the LAB strains from a specialty pig (Bamei pig) feces of the Tibetan plateau region and determine their antimicrobial properties and environmental tolerance to evaluate their potential probiotic values. A total of 116 LAB strains were isolated, from which the LAB strain Qinghai (QP)28-1 was identified as Lactiplantibacillus (L.) plantarum subsp. plantarum using 16S rDNA sequencing and recA amplification, showing the best growth capacity, acid production capacities, environmental tolerance, hydrophobicity, antibiotic susceptibility, and bacteriocin production capacity. Furthermore, this strain inhibited the growth of multiple pathogens by producing organic acids and bacteriocin. These bacteriocin-encoding genes were identified using PCR amplification, including plnS, plnN, and plnW. In conclusion, bacteriocin-producing L. plantarum subsp. plantarum QP28-1 stands out among these 116 LAB strains, and was considered to be a promising strain used for LAB-related food fermentation. Moreover, this study provides a convenient, comprehensive, and shareable profile for screening of superior functional and bacteriocin-producing LAB strains, which can be used in the food industry.

Dynamics Changes of Microorganisms Community and Fermentation Quality in Soybean Meal Prepared with Lactic Acid Bacteria and Artemisia argyi through Fermentation and Aerobic Exposure Processes.

This study evaluated the effects of Lactiplantibacillus plantarum subsp. plantarum ZA3, Artemisia argyi and their combination, on the fermentation characteristics, microbial community, mycotoxins and crude flavonoids content of fermented soybean meal during fermentation (under anaerobic conditions) and aerobic exposure (under aerobic conditions). The results showed that ZA3, Artemisia argyi and ZA3+ Artemisia argyi groups had lower pH values and higher lactic acid concentrations compared with controls, and additives increased the abundance of Lactiplantibacillus and decreased those of Acetobacter and Enterobacter; in particular, Artemisia argyi and ZA3+ Artemisia argyi reduced the abundance of fungi, such as Aspergillus, Pichia, Fusarium, Cladosporium and Xeromyces. Meanwhile, the contents of mycotoxins were lower in treated groups, and even mycotoxins in the control were significantly reduced after 30 d (p < 0.05). Crude flavonoids that were correlated positively with Lactococcus and negatively with Bacillus, Aspergillus, Enterobacter and Kazachstania were significantly higher in the Artemisia argyi and ZA3+ Artemisia argyi groups (p < 0.05).

Probiotic Potential and Wide-spectrum Antimicrobial Activity of Lactic Acid Bacteria Isolated from Infant Feces.

In this study, we aimed to characterize lactic acid bacteria strains derived from infants' feces, to evaluate the probiotic potential and explore the wide-spectrum antimicrobial activity. Of 800 isolates, 20 inhibited the growth of enterotoxigenic Escherichia coli K88 and Salmonella enterica ATCC 13076. On the basis of 16S rRNA sequence analysis, the 20 isolates were assigned to Lactobacillus casei (7), Lactobacillus paracasei (2), Lactobacillus plantarum (4), Lactobacillus rhamnosus (2), Enterococcus avium (3), Enterococcus faecium (1), and Enterococcus lactis (1) species. In addition, 12 strains with high antimicrobial activity were investigated for the presence of probiotic properties such as physiological-biochemical characteristics, antimicrobial susceptibility, hemolytic activity, hydrophobicity, and aggregation activity. Wide-spectrum antimicrobial activity analysis revealed that approximately all tested strains inhibited the ten pathogens, and four strains (ZX221, ZX633, ZX3131, and ZX3875) had good probiotic properties and survived after being exposed to simulated gastrointestinal tract conditions. Moreover, we investigated the influence of pH on the wide-spectrum antimicrobial activity and found that four strains inhibited most pathogens at pH 4.5 and pH 5, whereas only ZX633 had an inhibitory effect on Bacillus subtilis ATCC 6633 and Micrococcus luteus ATCC 4698 at pH 5.5. Overall, Lact. casei ZX633 had wide-spectrum antimicrobial activity and could be considered a potential probiotic.

Screening of Lactic Acid Bacteria with Inhibitory Activity against ETEC K88 as Feed Additive and the Effects on Sows and Piglets.

Enterotoxigenic Escherichia coli (ETEC), which expresses K88 is the principal microorganism responsible for bacterial diarrhea in pig husbandry, and the indiscriminate use of antibiotics has caused many problems; therefore, antibiotics need to be replaced in order to prevent diarrhea caused by ETEC K88. The objective of this study was to screen excellent lactic acid bacteria (LAB) strains that inhibit ETEC K88 and explore their effects as probiotic supplementation on reproduction, growth performance, diarrheal incidence, and antioxidant capacity of serum in sows and weaned piglets. Three LAB strains, P7, P8, and P15, screened from 295 LAB strains and assigned to Lactobacillus (L.) reuteri, L. amylovorus, and L. johnsonii with high inhibitory activity against ETEC K88 were selected for a study on feeding of sows and weaned piglets. These strains were chosen for their good physiological and biochemical characteristics, excellent exopolysaccharide (EPS) production capacity, hydrophobicity, auto-aggregation ability, survival in gastrointestinal (GI) fluids, lack of hemolytic activity, and broad-spectrum activity against a wide range of microorganisms. The results indicate that LAB strains P7, P8, and P15 had significant effects on improving the reproductive performance of sows and the growth performance of weaned piglets, increasing the activity of antioxidant enzymes and immune indexes in both.

Effects of Lactobacillus plantarum on the Fermentation Profile and Microbiological Composition of Wheat Fermented Silage Under the Freezing and Thawing Low Temperatures.

The corruption and/or poor quality of silages caused by low temperature and freeze-thaw conditions makes it imperative to identify effective starters and low temperature silage fermentation technology that can assist the animal feed industry and improve livestock productivity. The effect of L. plantarum QZ227 on the wheat silage quality was evaluated under conditions at constant low temperatures followed by repeated freezing and thawing at low temperatures. QZ227 became the predominant strain in 10 days and underwent a more intensive lactic acid bacteria fermentation than CK. QZ227 accumulated more lactic acid, but lower pH and ammonia nitrogen in the fermentation. During the repeated freezing and thawing process, the accumulated lactic acid in the silage fermented by QZ227 remained relatively stable. Relative to CK, QZ227 reduced the abundance of fungal pathogens in silage at a constant 5°C, including Aspergillus, Sporidiobolaceae, Hypocreaceae, Pleosporales, Cutaneotrichosporon, Alternaria, and Cystobasidiomycetes. Under varying low temperature conditions from days 40 to days 60, QZ227 reduced the pathogenic abundance of fungi such as Pichia, Aspergillus, Agaricales, and Plectosphaerella. QZ227 also reduced the pathogenic abundance of Mucoromycota after the silage had been exposed to oxygen. In conclusion, QZ227 can be used as a silage additive in the fermentation process at both constant and variable low temperatures to ensure fast and vigorous fermentation because it promotes the rapid accumulation of lactic acid, and reduces pH values and aerobic corruption compared to the CK.

Antibacterial Activity of Lactic Acid Producing Leuconostoc mesenteroides QZ1178 Against Pathogenic Gallibacterium anatis.

Lactic acid bacteria (LAB) convert carbohydrates into organic acids [mainly lactic acid (LA)], which reportedly have bactericidal activities. Gallibacterium anatis is a Gram-negative bacteria which infects birds, and causes significant economic losses. In this study, we investigated the antibacterial activity of the LA producing, Leuconostoc mesenteroides QZ1178 from Qula (fermented food), against G. anatis, using the Oxford cup method. Our data showed that L. mesenteroides QZ1178 inhibited G. anatis isolates from different origins; however, L. mesenteroides QZ1178 antibacterial activity dropped dramatically at pH 5.5-pH 6. The LA concentration and pH of the liquid broth containing L. mesenteroides QZ1178 after 24 h culture was 29 mg/mL and 3.6, respectively. This concentration (29 mg/mL at pH 3.6) and the antibiotic, cefotaxime (minimum inhibitory concentration (MIC) 2.5 µg/mL) effectively inhibited G. anatis (GAC026) growth as observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Gallibacterium anatis treated with LA exhibited extensive cell surface collapse, increased cell damage, cell membrane disruption, and cytoplasmic leakage, indicative of cell lysis. We suggest L. mesenteroides QZ1178 exerts potential antibacterial effects against the poultry pathogen, G. anatis via LA.

Multilocus Sequence Typing of Leuconostoc mesenteroides Strains From the Qinghai-Tibet Plateau.

Leuconostoc mesenteroides strains were a type of epiphytic bacterium widely used in fermented foods and products in the biochemical and pharmaceutical industries but data on its presence in foods from Qinghai-Tibet Plateau in China was scarce. In this study, molecular analysis based on multilocus sequence typing (MLST) with eight housekeeping genes (pyrG, groeL, rpoB, recA, uvrC, murC, carB, and pheS) was carried out on 45 L. mesenteroides strains isolated from different plants and dairy products from Qinghai-Tibet Plateau in China. The objective of this study was to perform genetic diversity analysis and explore the relationship between strains and isolate samples or separate regions. A total of 25 sequence types (STs) were identified with a diversity of up to 55.6%, which were grouped into one clonal complexes (CCs), 3 doublets and 17 singletons by eBURST. The results of minimum spanning tree and clustering analysis indicated these L. mesenteroides strains from the Qinghai-Tibet Plateau were relatively weakly related to the isolated region. However, there was a close relationship between the genotypes of L. mesenteroides strains and the type of the isolated sample, which was consistent with the results of API 50CH. The MLST scheme presented in this study provides a shareable and comparable sequence database and enhances our knowledge of the population diversity of L. mesenteroides strains which will be further used for the selection of industrial strains.

The in vitro Effects of the Probiotic Strain, Lactobacillus casei ZX633 on Gut Microbiota Composition in Infants With Diarrhea.

We investigated the in vitro effects of Lactobacillus casei ZX633 on gut microorganism composition in infants with diarrhea. For this purpose, 103 feces samples from healthy infants (healthy group) and 300 diarrhea samples from infants (diarrhea group) were collected, and diarrhea feces were treated with L. casei ZX633, which was previously isolated from healthy infant feces (treatment group). We used microbial dilution plate methods, high performance liquid chromatography (HPLC) and high-throughput sequencing approaches to analyze viable main microorganism counts, short chain fatty acid (SCFA) concentrations, and intestinal microbiota composition in feces, respectively. Our data showed that L. casei ZX633 supplementation increased the numbers of Escherichia coli, yeasts, lactic acid bacteria (LAB) and aerobic-bacteria, raised propionic acid levels but reduced four other SCFAs, which are close to the healthy group. Alpha diversity results indicated that microbial diversity and richness decreased in treatment group. Bacterial community analyses revealed that microbial structures of the treatment group tended toward the healthy group; i.e., Escherichia-Shigella and Clostridioides abundance increased, and there was a reduction in the abundance of Streptococcus, Bacteroides, Enterococcus and Veillonella. In conclusion, L. casei ZX633 isolated from healthy infant feces, may be effective in improving infant diarrhea microbiota, potentially providing a new probiotic strain to reduce the incidence of diarrhea associated with bacterial disease in infants.