Isolation and Identification of Cellulase-producing Bacillus in Yak Rumen

To investigate the species of cellulase-producing Bacillus in yak rumen,10 samples of rumen content were aseptically collected from 10 adult Maiwa yaks to isolate the heat-resistant Bacillus by water bath at 80 ℃ for 20 min.The cellulase-producing strains were screened using the CMC-Na medium and Congo red staining.The 16S rRNA gene sequence of those cellulose-producing strains were amplified and sequenced.The results showed that 64 strains were isolated from the 10 samples.Total 23 strains were identified as cellulase-producing bacillus,including 16 strains of Bacillus cereus,7 strains of Bacillus thuringiensis.Furthermore,phylogenetic analysis showed that the 16 Bacillus cereus strains were clustered into two branches:One isolate was clustered into a branch alone,the other 15 isolates were clustered into a branch which clustered into 5 small branches,showing that there was certain genetic diversity in the isolates of Bacillus cereus.And all 7 Bacillus thuringiensis strains were clustered into a branch.Hence,the results layed the foundation of investigating the species of cellulase-producing Bacillus in yak rumen and developing probiotics special for yak.     

Screening of Probiotics Inhibiting E. coli K88 in vitro

The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.     

Research Lactobacillus rhamnosusFermented Herbal and Bacillus subtilis Complex on Immune Performance,E.coli Infection of Broiler

This experiment was designed to study the complex of Lactobacillus rhamnosus fermented herbal and Bacillus subtilis on White Feather broiler immunity performance and impact of Escherichia coli infection.360 one-day-old broiler chickens were randomly divided into 3 groups with 4 replicates in each group and 30 chickens per replicate.The pretrial period lasted for 7 d,and the experiment lasted for 35 d.The chickens in the group Ⅰ(positive control group) and group Ⅱ(negative control group)were all only fed a basal diet,group Ⅲ was test group,by additive 1% fermented herbal preparations,groups Ⅱ and Ⅲ were intraperitoneal injection of 1 mL E.coli at 35 d,broiler mortality,immune organ index,cecalmicroflora content,immunoglobulin levels,IL-2 and IL-6 content were tested.The results showed that injection of E.coli caused massive death of chickens,group Ⅱtook up to 75.00%,it was significantly higher than groups Ⅰ and Ⅲ (P < 0.05),the mortality in group Ⅲ was significantly lower than group Ⅱ(P < 0.05),was only 23.33%.Injection of E.coli maked spleen index and thymus index of group Ⅱincreased significantly (P < 0.05),the spleen index and thymus index of groups Ⅰ and Ⅲ were no significant difference (P > 0.05).E.coli counts was significantly decreased after injectionin group Ⅲ (P < 0.05),but the number of intestinal Lactobacilli of group Ⅲ was significantly increased (P < 0.05),and inhibited the propagation of E.coli,the counts of E.coli in groups Ⅰ and Ⅲ were no significant difference (P > 0.05).At 42 d,the sIgA of the intestinal fluid in group Ⅲ were higher than that of groups Ⅰ and Ⅱ with 11.99% and 36.56%,respectively(P < 0.05).The serum IgG concentrations of group Ⅲ was higher than that of groupsⅠand Ⅱwith 14.68% and 28.15%,respectively(P < 0.05).At 42 d,the IL-2 content of group Ⅱ was the lowest,it was significantly lower than group Ⅲ(P < 0.05),the IL-6 of group Ⅲ was significantly lower than group Ⅱ(P < 0.05).     

凝结芽孢杆菌对热应激肉鸡生长性能及肠道消化与吸收功能的影响

本试验旨在研究凝结芽孢杆菌对热应激肉鸡生长性能及肠道消化与吸收功能的影响。选择320只28日龄健康爱拔益加（AA）肉公鸡,随机分为5个组,每组8个重复,每个重复8只。常温对照组（TC组）肉鸡饲养于（22±1）℃的常温环境,饲喂基础饲粮;热应激组（HS组）肉鸡饲养于（34±1）℃的高温环境,饲喂基础饲粮;热应激试验组肉鸡饲养于（34±1）℃的高温环境,分别饲喂在基础饲粮中添加0.2×107(HS-BC0.2组)、1.0×107(HS-BC1.0组)、5.0×107CFU/g(HS-BC5.0组)凝结芽孢杆菌的试验饲粮。试验期14 d。结果表明：与TC组相比,HS组的末重、平均日增重（ADG）、平均日采食量（ADFI）,干物质和粗蛋白质表观代谢率,空肠食糜总淀粉酶和总蛋白酶酶活,空肠绒毛高度、绒毛高度/隐窝深度以及空肠钠/葡萄糖协同转运蛋白1(SGLT1)、基础氨基酸转运载体（r-BAT）、y+L氨基酸转运载体-1(y+LAT1) mRNA表达量均显著降低（P<0.05）,料重比（F/G）显著增加（P<0.05）。与HS组相比,HS-BC1.0和HS-BC5.0组的ADG、粗蛋...     

紫外辐照对产α-ALDC枯草芽孢杆菌的诱变效应

采用紫外诱变(波长260nm，功率15w)对一株产α-乙酰乳酸脱羧酶的枯草芽孢杆菌（Bacillus subtilis）BS059菌株进行诱变处理。结果表明，在紫外辐照时间为50s时，诱变效果较好。从正突变菌株中反复筛选，得到两株产酶量较高的菌株BS059-15和BS059-22，比原出发菌株的酶活分别提高了107.62%和162.25%。经过连续传代试验，证明其遗传性状稳定，为可遗传变异。     

猕猴桃溃疡病生防芽孢杆菌B2的鉴定及应用

猕猴桃溃疡病是一种毁灭性细菌病害,该病发展迅速,防治难度大,严重制约了猕猴桃产业的发展。为找到一种有效防治该病的方法,针对化学防治易产生抗药性、污染环境、杀伤天敌等特点,从猕猴桃根际土壤中分离共获得25株芽孢杆菌,采用点菌法筛选出4株对猕猴桃溃疡病菌(Pseudomonas syringae pv.actinidiae)具有拮抗效果的菌株(抑菌圈直径达到5 mm以上),其中B2拮抗作用最强。根据菌落(体)形态特征、生理生化鉴定以及16S r DNA序列分析,初步鉴定B2为蜡样芽孢杆菌(Bacillus cereus)。将B2的发酵原液稀释成1×109cfu/m L的发酵液,并在此基础上依次配制50、100、500、1000、2000倍的稀释液,用于盆栽试验。盆栽试验采用喷雾法,30天后的防治效果分别达到了90.45%、86.52%、80.90%、67.98%、47.19%、15.73%,60天后尽管病斑有少量增加,旧病斑有所增大,但与对照相比,病斑增长速度明显减缓。大田试验结果证明,该菌具有较好的生防潜力,喷雾法选择B2发酵液的50倍稀释液,涂抹法选择B2发酵液的100倍稀释液,可有效降低猕猴桃溃疡病的危害。     

柑橘溃疡病拮抗芽孢菌株的筛选及其控病作用

从柑橘根际土壤中获得192个细菌分离物,通过平板拮抗测试,筛选出对柑橘溃疡病菌（Xanthomonas axonopdis pv. citri）有较强拮抗作用的6个芽孢菌株,抑菌圈直径达12.0～25.0mm,其中菌株G32对柑橘溃疡病菌抑制效果最好,稳定性高;温室控病试验表明,菌株G32的控病效果最佳,提前喷施拮抗菌其预防效果达60.7%,接种溃疡病菌后喷施的治疗效果为53.2%;根据其形态特征、培养性状、生理生化特性及16S rDNA序列比对分析,将G32鉴定为枯草芽孢杆菌Bacillus subtilis。     

一株马铃薯干腐病生防菌的鉴定及其抑菌活性研究

镰孢菌引起的马铃薯干腐病是影响马铃薯贮存期间产量与品质的重要病害之一。寻找高效低毒的防治马铃薯干腐病的生防药剂对提高马铃薯的产量与质量具有重要意义。本研究以腐皮镰孢Fusarium solani为病原菌, 测定分离自马铃薯块茎的菌株2-1-9-CJK-2的抑菌活性。该菌株发酵液、无菌发酵液、菌悬液、胞内组分以及挥发性有机物对腐皮镰孢均具有抑菌活性, 抑制率分别为93.37%、61.92%、80.82%、44.01%和49.16%。结合形态学、生理生化特征和多基因系统发育树构建鉴定其为贝莱斯芽胞杆菌Bacillus velezensis。马铃薯切片试验和马铃薯块茎试验表明, 当菌株发酵液浓度为1×109 cfu/mL时, 对病原菌的抑制效果最好。除此之外, 菌株可以分泌蛋白酶、淀粉酶、纤维素酶, 具有溶磷能力和生物膜形成能力, 但不具有溶血性, 并且对供试的其他6种病原真菌有较强的抑菌活性。研究表明贝莱斯芽胞杆菌2-1-9-CJK-2抑菌谱广, 具有生物安全性, 对腐皮镰孢有较强的抑菌活性, 在植物病害防治中具有一定的应用潜力, 可为马铃薯干腐病生防制剂的开发奠定基础。