Oral administration of Bacillus coagulans TQ-35 alleviates allergic responses in OVA-sensitive BALB/c mice

Bacillus coagulans has been extensively studied so far,but there has been a lack of research on its usage in allergy.In this study,we designed to assess the effect of different concentrations of B.coagulans on food allergy in a BALB/c mouse model of ovalbumin(OVA)-induced food allergy and its effect on gut microbes.The assessment of symptoms,specific immunoglobulin E(IgE),T-cell differentiation,and related gene expression levels in sensitized mice by assay indicated that high doses of oral B.coagulans could alleviate allergic symptoms.Treatment with B.coagulans,in the high-dose group,significantly reduced IgE and IgG1 levels and modulated the balance of T helper type 1 cell(Th1)and Th2 and the expression of relevant genes in the spleen.16S rRNA analysis showed that probiotics improved the structure of the microbiota,in particular by boosting the percentage of Clostridia,Bacteroides vulgatus and Enterococcus faecium,and by increasing the abundance of microbial species,thereby modulating the immune system.Therefore,this study can provide insights into the practical application of B.coagulans doses to alleviate OVA allergy.

The synbiotic mixture of lactulose and Bacillus coagulans protects intestinal barrier dysfunction and apoptosis in weaned piglets challenged with lipopolysaccharide

Background Lactulose as an effective prebiotic protects intestinal mucosal injury.Bacillus coagulans is widely used in feed additives because of its ability to promote intestinal health.Our previous study suggests that the combination of lactulose and Bacillus coagulans may be a good candidate as alternative for antibiotic growth promoters.However,the in vivo effects of lactulose and Bacillus coagulans on growth and intestinal health under immune challenge in piglets remains unclear.The objective of this study is to explore the protective effects of synbiotic containing lactu-lose and Bacillus coagulans on the intestinal mucosal injury and barrier dysfunction under immune challenge in weaned piglets.Methods Twenty four weaned piglets were assigned to 4 groups.Piglets in the CON-_(saline)and LPS-_(LPS)group were fed the basal diet,while others were fed either with chlortetracycline(CTC)or synbiotic mixture of lactulose and Bacillus coagulans for 32 d before injection of saline or lipopolysaccharide(LPS).Piglets were sacrificed 4 h after LPS injection to collect samples to determine intestinal morphology,integrity and barrier functions as well as relative genes and proteins.Results Our data showed that no differences were observed in the growth performance of the four test groups.LPS injection induced higher serum diamine oxidase activities,D-lactic acid levels,and endotoxin status,lower villus height and ratio of villus height to crypt depth,greater mRNA and lower protein expression related tight junction in both jejunum and ileum.In addition,a higher apoptosis index,and protein expression of Bax and caspase-3 were also observed in the LPS challenge group.Interestingly,dietary synbiotic mixture with lactulose and Bacillus coagulans protected against LPS-induced intestinal damage,barrier dysfunction and higher apoptosis as well as CTC.Conclusions Our data suggest that dietary supplementation of synbiotic mixture with lactulose and Bacillus coagu-lans showed resilience to LPS-induced intestinal morphological damage,barrier dysfunction and aggressive apoptosis in piglets as well as the protective effects of CTC.These results indicate that synbiotic mixture of lactulose and Bacillus coagulans showed beneficial effects on performance and resilience to acute immune stress in weaned piglets.

Mechanism of the Effect of Bacillus coagulans on Production Performance of Laying Hens in Late Laying Period

[Objective]The paper was to investigate the mechanism of Bacillus coagulans preparation in improving production performance of laying hens in late period of laying.[Method]A total of 648 individuals of"Jingfen 1"laying hens(66 weeks of age)in late period of laying were randomly divided into four groups,six replicates each group,and each group had 27 individuals of laying hens.The laying hens in control group were fed with basal diet,and those in experimental groupsⅠ,Ⅱ and Ⅲ were fed with the basal diets added with 3.33×10^6,1×10^7,3.33×10^7 CFU/g B.coagulans,respectively.The pretrial lasted one week,and the formal test lasted eight weeks.[Result]Compared with the control group,the laying rate in group Ⅰ increased significantly,and the feed-gain ratio in experimental group Ⅱ decreased noticeably while the spleen index increased remarkably;the alkaline phosphatase activities and blood calcium content in experimental groups Ⅰ,Ⅱ and Ⅲ increased significantly(P<0.05),while the triglyceride content decreased remarkably(P<0.05);the urea content in experimental groups Ⅰ and Ⅱ decreased obviously(P<0.05).Adding B.coagulans significantly increased the specific activity of amylase,lipase and protease in various intestinal mucosa of small intestine(P<0.05);adding B.coagulans significantly increased the content of follicle-stimulating hormone(FSH)and estradiol(E2),and significantly increased the mRNA expression of follicle stimulating hormone receptor gene(FSHR).[Conclusion]B.coagulans preparation could significantly improve the production performance of laying hens in late period of laying;appropriately enhance the immune capacity of laying hens;improve the serum biochemical indicators;increase the activity of digestive enzymes in the small intestine;and promote the release of gonadal hormone and the mRNA expression of FSHR gene in the ovary.

MLST analysis of genetic diversity of Bacillus coagulans strains to evaluate effects on constipation model

Bacillus coagulans can help ameliorate or prevent gastrointestinal diseases, but the genetic relationships among B. coagulans isolates are not well studied. Multilocus sequence typing analysis was conducted on 57 isolates of B. coagulans from 22 provinces or autonomous regions in China. B. coagulans isolates were highly diverse and a total of 33(sequence typings) STs were found. These isolates had a weak clonal population structure and strong indications of intraspecies recombination. The evolution direction of B. coagulans was not correlated with geography or isolation source. Fifteen strains were selected for further analysis based on proximity relationships from the phylogenetic tree. Five isolates(B. coagulans-1, B. coagulans-10, B. coagulans-39, B. coagulans-70 and B. coagulans-71) with good spore-forming ability relative to the rest of the isolates were evaluated for constipation relief. B. coagulans-39 significantly relieved constipation symptoms in mice by regulating intestinal flora, increasing the production of short-chain fatty acids and restoring the level of gastrointestinal regulatory peptides. Comparative genomic analysis showed the beneficial effects of B. coagulans-39 might be associated with specific functional genes that are involved in the utilization of various carbohydrates as primary substrates and short-chain fatty acid production.

Alleviative effects of a novel strain Bacillus coagulans XY2 on copper-induced toxicity in zebrafish larvae

Copper (Cu) is a kind of micronutrient element that is essential for human metabolism.However,it is also considered as an environmental pollutant which is toxic to organisms at a high concentration level.Probiotics,regarded as beneficial microorganisms for promoting human health,have functions of antioxidant capacity,immune-enhancing properties,intestinal barrier protection and regulation.Several studies have reported that probiotics show positive effects on alleviating and intervening heavy metals toxicity.However,evidence for relieving copper-induced toxicity by probiotics is still limited.In this study,we firstly conducted a zebrafish larvae model to screen out microorganisms which are helpful for CuSO_(4)toxicity resistance and one novel strain named as Bacillus coagulans XY2 was discovered with the best protective activity.B.coagulans XY2 significantly reduced the mortality of zebrafish larvae exposed to 10μmol/L CuSO_(4)for 96 hr,as well as alleviated the neutrophils infiltration in the larvae lateral line under a 2 hr exposure.B.coagulans XY2 exhibited a high in vitro antioxidant activity and against CuSO_(4)-induced oxidative stress in zebrafish larvae by up-regulating sod1,gstp1 and cat gene transcriptional levels and relevant enzymatic activities.CuSO_(4)stimulated the inflammation process resulting in obvious increases of gene il-1βand il-10 transcription,which were suppressed by B.coagulans XY2 intervention.Overall,our results underline the bio-function of B.coagulans XY2 on protecting zebrafish larvae from copper toxicity,suggesting the potential application values of probiotics in copper toxicity alleviation on human and the environment.

Effects of benzoic acid,Bacillus coagulans and oregano oil combined supplementation on growth performance,immune status and intestinal barrier integrity of weaned piglets

This experiment was conducted to investigate the effects of benzoic acid,Bacillus coagulans and oregano oil combined supplementation on growth performance,immune status and intestinal barrier integrity of piglets.In a 26-d experiment,25 piglets were randomly assigned to 5 treatments:1)a basal diet,negative control(NC),2)NC added with antibiotics,positive control(PC);3)NC added with benzoic acid at 3,000 g/t and Bacillus coagulans at 400 g/t(AB);4)NC added with benzoic acid at 3,000 g/t and o regano oil at 400 g/t(AO);5)NC added with 3,000 g/t benzoic acid and Bacillus coagulans at 400 g/t and oregano oil at 400 g/t(ABO);On d 27,all piglets were euthanized to obtain jejunal mucosa to measure immune status and intestinal barrier integrity.Results showed that pigs fed AB diet increased the final body weight and average daily body weight gain and decreased the ratio of feed to gain co mpared with NC group(P<0.05).Co mpared with NC group,AB,AO and ABO decreased serum tumor necrosis factor-a concentration and ABO decreased interleukin-1βconcentration in serum and jejunal mucosa(P<0.05).Compared with NC group,AB upregulated mRNA expressions of sodium-glucose cotransportel,claudin-1,occludin and mucin2 in jejunal mucosa and the populations of Bifidobacterium and Bacillus in cecal digesta(P<0.05).Compared with NC group,ABO increased jejunal mucosal occludin mRNA abundance and Bifidobacterium population in cecal digesta,and decreased Escherichia coli population in cecal digesta(P<0.05).Furthermore,AB and ABO increased Bacillus population in cecal digesta compared with PC group(P<0.05).These results indicated that dietary AB supplementation could improve growth performance and intestinal barrier integrity of piglets when fed antibiotic-free diets,which was possibly associated with the improvement of immune status and intestinal microflora.Dietary ABO supplementation is also beneficial to improve immune status and intestinal barrier integrity and micro flora of piglets.

凝结芽胞杆菌发酵优化工艺

为提高凝结芽胞杆菌(Bacillus coagulans)的生长效率,在摇瓶、发酵罐水平考察了最适的碳源、温度、维生素添加量、pH以及发酵过程存在的氧化胁迫对凝结芽胞杆菌生长代谢的影响,获得了发酵培养基配方,并确定了30 L发酵罐最适培养条件:培养温度为40℃,初始pH为6.2~6.5,培养过程中9~14 h用氨水溶液控pH=5.5,pH大于5.5后停止控制,全程以4.1 mg/(L·h)速率流加半胱氨酸。优化条件下,发酵13 h细胞生物量达1.9×10^(10)CFU/mL,较对照提高363.4%,细胞生长效率为1.5×10^(9) CFU/(mL·h),较对照提高305.4%。显著提高了生物量、缩短了发酵周期,为凝结芽胞杆菌工业化生产提供了重要参考。

凝结芽孢杆菌BC320对南美白对虾生长和血液免疫指标的影响

为验证一株分离自南美白对虾体内的凝结芽孢杆菌(Bacillus coagulans)BC320对南美白对虾(Penaeus vannamei)生长性能和非特异性免疫指标的影响,将1200尾南美白对虾(平均体重3.24 g±0.2 g)随机分成4组,分别添加0‰、0.5‰、1‰、1.5‰的BC320,每组300尾,3个重复,每个重复100尾。经40 d养殖后,检测平均增重率(AWG)、特定生长率(SGR)、饲料系数(FC)和成活率(SR)等生长指标和超氧化物歧化酶(SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)、溶菌酶(LSM)、过氧化物酶(POD)等免疫指标。结果显示,终末体重、增重率和特定生长率指标,各处理组与对照组相比均有显著增加,但在添加1‰和1.5‰之间没有显著性差异;当添加量为1‰时,试验组的SOD、ACP、ALP和POD与对照组均有显著差异,LSM与对照组之间没有显著差异。本试验结果表明,凝结芽孢杆菌BC320可促进南美白对虾生长,提高其非特异性免疫水平。

不同来源寡聚-1,6-葡糖苷酶异源表达及酶学特性

寡聚-1,6-葡糖苷酶(EC 3.2.1.10)对于乳酸发酵液残糖中含量较高的异麦芽糖有较高的利用活性。基于前期研究工作基础并经BRENDA数据库搜索挑选了5个可能耐高温、耐酸的寡聚-1,6-葡糖苷酶基因(Bacillus coagulans ATCC 7050的malL BF292011(BC1)和malL BF292004(BC2)、Bacillus subtilis 168的malL yvdL BSU34560(BS1)和yugT BSU31290(BS2)以及Bacillus thermoglucosidasius KP 1006的malL(KP)),首次利用pET表达系统在大肠杆菌中进行重组表达、纯化及酶学性质的系统研究。结果显示:BC1、BC2、KP的最适pH为5.0,BS1、BS2的最适pH为6.0。最适温度分别为:KP(60℃),BC1(56℃),BC2(56℃),BS1(43℃),BS2(43℃)。Mg^(2+)、NH_(4)^(+)对于BC1的酶催化活性具有激活作用,Co^(2+)、K+、NH_(4)^(+)对于KP有激活作用,但这些离子对于BC2、BS1和BS2都没有明显的激活作用。Zn^(2+)、Cu^(2+)、Ni^(+)对这5种寡聚-1,6-葡糖苷酶都有明显的抑制作用。以对硝基苯-α-D-葡萄糖苷(pNPG)为底物时,酶的转换数和催化效率从高到低依次为:KP>BC1>BS1>BS2>BC2。以乳酸发酵液中的残糖成分作为参考,在以麦芽糖、异麦芽糖、蔗糖、乳糖、海藻糖以及可溶性直链淀粉作为底物测定酶活时,BC1、BC2、KP、BS1的最适底物都是异麦芽糖,BS2的最适底物为麦芽糖。

Combination eect of pH and acetate on enzymatic cellulose hydrolysis

The productivity and efficiency of cellulase are significant in cellulose hydrolysis. With the accumulation of volatile fatty acids （VFAs）, the pH value in anaerobic digestion system is reduced. Therefore, this study will find out how the pH and the amount of acetate influence the enzymatic hydrolysis of cellulose. The effects of pH and acetate on cellulase produced from Bacillus coagulans were studied at various pH 5-8, and acetate concentrations （0-60 mmol/L）. A batch kinetic model for enzymatic cellulose hydrolysis was constructed from experimental data and performed. The base hypothesis was as follows： the rates of enzymatic cellulose hydrolysis rely on pH and acetate concentration. The results showed that the suitable pH range for cellulase production and cellulose hydrolysis （represents efficiency of cellulase） was 2.6-7.5, and 5.3-8.3, respectively. Moreover, acetate in the culture medium had an effect on cellulase production （KI = 49.50 mmol/L, n = 1.7） less than cellulose hydrolysis （/（＇i = 37.85 mmol/L, n = 2.0）. The results indicated that both the pH of suspension and acidogenic products influence the enzymatic hydrolysis of cellulose in an anaerobic environment. To enhance the cellulose hydrolysis rate, the accumulated acetate concentration should be lower than 25 mmol/L, and pH should be maintained at 7.

一株具有降解玉米赤霉烯酮能力的凝结芽孢杆菌的分离、鉴定及其发酵条件的确定

从池塘养殖鲤鱼中分离出一株具有降解玉米赤霉烯酮能力的芽孢杆菌,经16SrDNA比对鉴定为凝结芽孢杆菌(Bacillus coagulans)。经液相色谱方法检测其发酵上清液对玉米赤霉烯酮的24、48h降解能力分别为62%和84%。经发酵实验得到此株凝结芽孢杆菌的最适生长条件为:40℃,pH值6.4,经36h好氧液体发酵可得25.9亿个/mL的活菌数,最佳液体培养基为:蛋白胨15‰,酵母粉5‰,玉米粉15‰,葡萄糖20‰,麸皮10‰。综合实验结果,此株凝结芽孢杆菌可作为饲料添加剂用于饲料中霉菌毒素的降解和提高动物抗病能力。

海岸水虱消化道的凝结芽孢杆菌分离鉴定及其产酶能力研究

采用传统的微生物富集、分离纯化方法,对海岸水虱(Ligia exotica)消化道微生物进行分离鉴定,得到1株产酶能力较强的菌株,命名为HZL-3。利用分子生物学方法,初步确定其为凝结芽孢杆菌(Bacillus coagulans)。采用ELISA法检测HZL-3产蛋白酶、淀粉酶含量,并研究温度、pH、培养时间、接种量等因素对其产酶量的影响。结果表明,HZL-3能够产生蛋白酶和淀粉酶;温度36℃,pH为7.5,培养16 h,接种量为6%,蛋白酶活性最高;温度36℃,pH为6.0,培养10 h,接种量为4%,淀粉酶活性最高,研究旨在揭示海岸水虱消化道微生物作为水产养殖益生菌的潜在应用。