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1.
广西猪源益生菌的分离鉴定与生物学特性研究   总被引:1,自引:0,他引:1  
为了解广西地区猪粪便与微生态发酵饲料中益生菌的存在情况及合理开发和利用猪源益生菌,本试验共采集了15份猪粪便样品和5份益生菌发酵饲料并对其进行益生菌的分离,研究益生菌的形态学特性、生理生化特性及16S rRNA分子序列,进行生长曲线、耐胆盐试验、温度敏感试验、模拟胃肠道耐受试验、耐药性试验、体外抑菌试验、小鼠安全性试验、体内/外抑菌试验,对分离菌株进行生物学特性的研究分析。结果显示,本试验共分离鉴定出6种、29株益生菌,分别为1株屎肠球菌(C2)、2株乳酸肠球菌(C1、C3)、3株植物乳杆菌(R20、R30、R37)、4株干酪乳杆菌(R41~R44)、7株枯草芽孢杆菌(K1~K7)和12株蜡样芽孢杆菌(Y1~Y12)。从中筛选出6株具有生长性能好、耐胆盐、耐高温、对胃肠道耐受、体外抑菌能力强的益生菌:乳酸肠球菌C1、屎肠球菌C2、植物乳杆菌R20、干酪乳杆菌R41、枯草芽孢杆菌K1和蜡样芽孢杆菌Y3。将筛选出的具有优良特性的菌株进行小鼠安全性试验,结果表明,在10~9 CFU/mL浓度下灌胃6株益生菌对小鼠是安全无毒害的。与此同时,C2、R20、R41、Y3能极显著提高小鼠日增重(P0.01),K1能显著提高小鼠日增重(P0.05)。体内抑菌试验结果表明,C2、R20、K1能降低沙门氏菌G21感染的小鼠的死亡率,有一定程度的体内抑菌作用。各项试验结果表明,R20和K1可作为猪源益生菌的选择菌株。  相似文献   

2.
为了研制优良的猪用益生菌制剂,试验对从猪粪便分离得到的7株益生菌(地衣芽孢杆菌S3、产朊假丝酵母Y1、德氏乳杆菌德氏亚种B1、粪肠球菌C1和C2、枯草芽孢杆菌S1和S2)进行胃酸耐受性、肝脏胆盐耐受性、环境温度耐受性以及对大肠杆菌和葡萄球菌的抑制效果研究。结果表明:7株益生菌均能够耐受仔猪胃酸和小肠中胆盐,除芽孢杆菌外其他分离菌株均不耐热,在生产应用时需要考虑选用低温加工法;除S3和Y1外,其他菌株(B1、C1、C2、S1、S2)均有明显的抑菌作用,以S2抑菌效果最强。说明B1、C1、C2、S1、S2可用于益生菌制剂的研制菌种。  相似文献   

3.
《中国兽医学报》2016,(12):2054-2060
为了获得适合生产的羊源益生菌株,本试验从健康成年小尾寒羊肠道内分离出5株菌落形态各异、生长良好的菌株。通过革兰染色、生理生化鉴定及16SrDNA分析,5株菌分别为M1(干酪乳杆菌)、M2(植物乳杆菌)、E1(粪肠球菌)、E2(屎肠球菌)、L1(枯草芽孢杆菌)。细菌生长曲线、耐酸、耐胆盐、抑菌试验等益生特性分析结果表明,M1、M2和L1具有生长速度快,耐酸、耐胆盐,明显抑制大肠杆菌、鸡白痢沙门菌和金黄色葡萄球菌等肠道常见致病菌的益生特性,可以作为研制羊用微生态制剂的候选菌株。  相似文献   

4.
猪源乳酸菌的分离、筛选、鉴定及产酸性能的研究   总被引:3,自引:0,他引:3  
试验旨在分离、筛选出具有强抗逆性、产酸能力强的同源菌株。从28日龄断奶健康仔猪粪便分离得到30株乳酸菌,应用体外筛选法,筛选出在pH值2.5处理2h后的菌株13株;将此13株菌株通过浓度为0.2%胆盐后剩7株。采用牛津杯法进行抑菌试验,从7株菌中筛选出对大肠杆菌、沙门氏菌有较强抑菌作用的4株,分别为R11、R15、R18、R22,对此4株菌株进行产酸试验验证,都表现出强的产酸能力。对这4株菌进行鉴定性试验,结果R11为屎肠球菌,R15为鸡肠球菌,R18为嗜酸乳杆菌,R22为口乳杆菌。  相似文献   

5.
为了挖掘鸡源益生菌的菌种资源,试验采用肠道内容物分离纯化、生长曲线和产酸曲线绘制、抑菌性能测定、生化试验、基因测序、同源性分析、遗传进化树构建的方法对太行鸡肠道益生菌的种属和生物学特性进行了研究。结果表明:从散养太行鸡肠道分离、纯化了6株纯化益生菌,将其编号为J1~J6,其中J1~J4增殖速度快,2~10 h可达到对数生长期;6株菌均有一定的产酸能力,其pH值最低可达到3.78;6株菌对大肠杆菌和沙门氏杆菌的抑菌圈直径均大于14.67 mm,大肠杆菌对J3极度敏感,沙门氏杆菌对J1、J2极度敏感;生化试验显示,J1~J6依次为罗伊氏乳杆菌、粪肠球菌、枯草芽孢杆菌、鼠李糖乳杆菌、粪肠球菌、蜡样芽孢杆菌;PCR扩增得到大小为1 500 bp的目的条带,菌株序列与GenBank比对的结果与生化试验结果一致;各菌株与其GenBank标准菌株的同源性为96%~99%;J2、J5与粪肠球菌标准株遗传距离较远,与标准株相比有显著遗传变异,其余菌株均与标准株遗传距离较近,其中J1和J4遗传距离最近,但J3、J6与其余菌株遗传距离均较远。说明鸡源益生菌J1、J2和J4具备作益生菌饲料添加剂菌种的特性。  相似文献   

6.
为了解青海省共和县牦牛肠道内益生菌的分布种类,筛选出能够抑制犊牛腹泻的益生菌株,采集20份健康牦牛的粪便样品,对粪便中乳酸菌进行分离和纯化,再结合形态学、生化试验及分子生物学方法鉴定。结果表明,从牦牛粪便中成功分离出7株乳酸菌,包括罗伊氏乳杆菌、植物乳杆菌、保加利亚乳杆菌、乳酸乳球菌、坚强肠球菌、海氏肠球菌和屎肠球菌各1株。本研究分离的7株乳酸菌,为后期抗牦牛腹泻益生菌产品的研究奠定了基础。  相似文献   

7.
为开发羊源乳酸菌菌种资源,从健康成年散养山羊瘤胃内容物中分离乳酸菌,经生化试验、基因测序以及同源性分析、构建遗传进化树等方法对分离出的乳酸菌进行鉴定,并通过耐酸和耐胆盐试验筛选抗逆性良好的乳酸菌。结果表明:从散养山羊瘤胃内容物中分离出11株乳酸菌,将其编号为R1~R9、R11和R12;其中R1、R2为杜氏肠球菌,R3、R7为乳酸肠球菌,R4、R5、R6、R8、R9为屎肠球菌,R11为鼠李糖乳杆菌,R12为莱菲氏肠球菌;R4(屎肠球菌JQ067694.1)在pH值为2时存活率超过80%,在0.9%浓度胆盐下,存活率超过30%,抗逆性优于其他菌株,生产应用潜力较好。  相似文献   

8.
为了确定牛奶中的益生菌种类,给奶牛微生态制剂益生菌菌种的选择提供参考,试验选取体况良好、无乳房炎、产奶性能优良的荷斯坦奶牛和新生犊牛,采集其新鲜奶样和粪样,应用MRS溴甲酚紫选择性培养基和革兰氏染色对分离株进行初步鉴定,提取菌株DNA检测16S rRNA基因并测序,对分离株进行分子生物学鉴定。结果表明:所得菌株中有球菌和杆菌,革兰氏染色呈阳性,得到的12株菌株分别为约氏乳杆菌2株、布氏乳杆菌4株、粪肠球菌5株和屎肠球菌1株。这些牛源益生菌可用于研制益生菌微生态制剂。  相似文献   

9.
通过革兰染色、菌落形态观察、生化分析鉴定、牛津杯抑菌试验对分离到的1株乳酸菌进行研究。结果显示,经16 S r DNA序列测定,比对分析该株乳酸菌与鼠李糖乳杆菌、乳酸乳球菌乳亚种、唾液乳杆菌、乳链球菌、哥伦比亚肠球菌和鼠乳杆菌高度同源。乳酸菌抑菌试验检测显示,对大肠杆菌、沙门菌、蜡样芽孢杆菌和金黄色葡萄球菌具有很强的抑菌效应。同时该株乳酸菌对水貂肠道上皮细胞有一定的粘附能力。结果表明,该株乳酸菌株产生菌能够粘附于水貂肠道上皮细胞上并具有较强的抑菌活性。  相似文献   

10.
研究采用MRS培养基从健康AA肉鸡十二指肠、空肠、回肠及盲肠中分离获得11株乳酸菌,并对其进行形态观察、生理生化试验及16S r DNA序列同源性分析,最终确定所分离的菌株主要是5种乳酸杆菌(嗜酸乳杆菌R1、植物乳杆菌R2、鼠李糖乳杆菌R3、短乳杆菌R4和发酵乳杆菌R5)和4种肠球菌(盲肠肠球菌R9、粪肠球菌R8和R10、鸟肠球菌R7、屎肠球菌R6和R11)。  相似文献   

11.
D. Taras  W. Vahjen  O. Simon 《Livestock Science》2007,108(1-3):229-231
Two probiotics of different ecological origin, Enterococcus faecium NCIMB 10415 and Bacillus cereus var. toyoi, were chosen as model organisms. Feed for sows during gestation/lactation and for piglets pre-/postweaning was supplemented with either of these probiotics. To evaluate the effect of different starting points of E. faecium NCIMB 10415 initiations the probiotic was administered to piglets of sows, which have not received the probiotic, from birth onwards or just postweaning. Here we report the impact of these variants on probiotic distribution in the gut, on the gut microbiota, on diarrhea and on performance. Both probiotic strains were detected immediately after the start of the supplementation in feces of sows and piglets. The vertical transfer of both probiotic strains with sow feces to piglets could be demonstrated already before suckling piglets had access to the supplemented diets. Both probiotics were recovered from all intestinal segments of piglets. The dominant autochthonous colonic microbiota of young piglets as revealed by denaturing gradient gel electrophoresis was more similar within than between treatment groups (control vs. probiotic). Both probiotics reduced the incidence of postweaning diarrhea (p < 0.05). For the E. faecium probiotic the relative magnitude of this effect was largely independent of dietary probiotic concentration or starting time of supplementation. Significant overall influence on piglet performance was observed only with the B. cereus probiotic.  相似文献   

12.
本研究以从断奶仔猪肠道分离的益生菌为试验菌株,将大肠杆菌K88和益生菌接种到体外培养的猪小肠上皮细胞(intestinal porcine epithelial cell-1,IPEC-1)中,测定培养2.5h上清液中的乳酸脱氢酶(lactate dehydrogenase,LDH)活性,同时将益生菌和大肠杆菌K88体外混合培养2.5h,进行平板计数,统计大肠杆菌K88菌数的变化,筛选出可以抑制大肠杆菌K88的益生菌。试验结果显示,干酪乳杆菌和凝结芽孢杆菌能显著降低IPEC-1培养上清液中的LDH活性(P<0.05),降低大肠杆菌K88对细胞的损伤;凝结芽孢杆菌能降低DMEM培养基中大肠杆菌K88的生长速度,结合模拟制粒过程及胃肠道环境的耐高温、耐酸及耐胆盐研究进行综合分析,该凝结芽胞杆菌对大肠杆菌K88有较好的抑制作用且具有良好的耐高温、耐酸及耐胆盐性能,具有作为微生态制剂菌株的应用潜力。本试验建立了能够抑制大肠杆菌K88的益生菌体外筛选技术模型。  相似文献   

13.
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.  相似文献   

14.
旨在研究单独添加屎肠球菌和枯草芽孢杆菌及二者混合添加对乳鸽生长性能、屠宰性能、免疫功能的影响,从而探究益生菌对乳鸽生长的调控机制。试验选取240只12日龄、体重接近的健康白羽王鸽,随机分成4组,分别为对照组、屎肠球菌组、枯草芽孢杆菌组、混合组,每组6个重复,每个重复10只乳鸽,试验期16 d。试验期结束时,每个重复随机选取2只乳鸽(共48只乳鸽)称重,测定其血清免疫指标、肌肉品质、肝组织抗氧化指标和生长相关基因表达量等。试验结果表明:1)与对照组相比,屎肠球菌组显著提高了乳鸽的平均日增重(P<0.05),且显著高于其余各组(P<0.05),而枯草芽孢杆菌组显著降低了乳鸽的平均日增重(P<0.05);2)与对照组相比,3个处理组对乳鸽的屠宰率、半净膛率、全净膛率、腿肌率、胸肌率、腹脂率均无显著影响(P>0.05),屎肠球菌组和混合组显著提高了乳鸽胸肌亮度(P<0.05),混合组还显著提高了乳鸽胸肌红度(P<0.05);3)与对照组相比,屎肠球菌组和混合组能显著提高乳鸽血清中IgG含量(P<0.05),混合组能显著提高乳鸽胸腺指数和法氏囊指数(P<0.05),除屎肠球菌组GLB显著高于对照组(P<0.05)外,3个处理组对乳鸽的其余血清生化指标均无显著影响(P>0.05);4)与对照组相比,屎肠球菌组超氧化物歧化酶(SOD)活性显著提高(P<0.05),混合组谷胱甘肽过氧化物酶(GSH-Px)活性显著提高(P<0.05);5)3个处理组肝组织的GHIGF-1基因mRNA表达水平均显著高于对照组(P<0.05),枯草芽孢杆菌组GHR基因mRNA表达水平显著高于对照组(P<0.05)。综上所述,屎肠球菌可显著提高乳鸽平均日增重,而枯草芽孢杆菌显著降低了乳鸽的平均日增重;屎肠球菌能改善乳鸽肌肉品质,提高乳鸽抗氧化性能和免疫功能,从而促进乳鸽生长。  相似文献   

15.
【目的】试验旨在研究植物乳杆菌GX20200417-1菌株的生物学特性及体外抑菌活性,探讨其在生产中应用的可能性。【方法】将植物乳杆菌GX20200417-1菌株接种至不同pH和含不同浓度胆盐的PBS,以及人工胃肠液中,使用菌落计数分析其对酸、胆盐和人工胃肠液的耐受性。使用牛津杯法检测GX20200417-1菌株对大肠杆菌、金黄色葡萄球菌及沙门氏菌的抑菌能力;通过与霉菌毒素共培养后使用ELISA方法研究GX20200417-1菌株对霉菌毒素的降解能力,并饲喂小鼠进行安全性试验。【结果】植物乳杆菌GX20200417-1菌株能够耐受pH 2.0及0.3%胆盐,并在人工胃肠液中有至少1×104 CFU/mL的活菌数;GX20200417-1菌株发酵上清液对大肠杆菌、沙门氏菌、金黄色葡萄球菌均有明显的抑制作用,对玉米赤霉素及黄曲霉毒素均有降解作用,但对呕吐毒素无降解作用;灌服GX20200417-1菌株后小鼠安全、无毒副作用。【结论】植物乳杆菌GX20200417-1菌株具有优良的益生特性,在畜禽生产中有一定的开发潜力。  相似文献   

16.
【目的】 探究干酪乳杆菌、嗜酸乳杆菌、乳双歧杆菌组成的复合益生菌对黄麻肉鸡生产性能及肠道微生物区系的影响, 为复合益生菌在肉鸡生产中的应用提供理论基础。【方法】 将200只1日龄雄性黄麻肉鸡随机分为2组, 每组5个重复, 每个重复20只。复合菌制剂(干酪乳杆菌∶嗜酸乳杆菌∶乳双歧杆菌=1∶1∶2)按1%比例补充于益生菌组肉鸡的饮用水中, 以正常饮水的肉鸡为对照。在42 d时以重复为单位测定肉鸡的生长性能指标, 并进行肠道菌群高通量测序分析。【结果】 与对照组相比, 益生菌组肉鸡的平均日增重显著增加(P<0.05), 平均日采食量、屠宰率和全净膛率增加, 料重比降低, 但差异不显著(P>0.05);益生菌组肠道菌群的OTU总数及回肠的Sob、Chao、Pd、Shannon、Shannoneven指数均降低, Simpson指数升高, 差异均不显著(P>0.05);厚壁菌门在肉鸡肠道中丰度最高, 具主体地位。较对照组而言, 益生菌组回肠中未见梭菌属, 厚壁菌门和乳球菌属的丰度均显著或极显著增加(P < 0.05; P < 0.01), 放线菌门和葡葡萄球菌属的丰度均显著或极显著降低(P < 0.05; P < 0.01), 盲肠中毛螺菌科的丰度显著增加(P<0.05)。【结论】 复合益生菌可通过增加肉鸡回肠乳杆菌属丰度、降低病原菌丰度来改善肠道菌群的组成、功能和多样性; 同时通过增加盲肠毛螺菌科丰度来提高肠道发酵降解非淀粉多糖的能力, 进而提高饲料利用效率, 促进肉鸡生长。  相似文献   

17.
为了解牦牛瘤胃中产纤维素酶的芽孢杆菌种类,无菌采集10份成年麦洼牦牛瘤胃内容物样本,经80 ℃高温水浴20 min后分离得到耐高温芽孢菌.将分离菌株点种至羧甲基纤维素钠培养基上,使用刚果红染液染色筛选产纤维素酶菌株,扩增产酶菌株16S rRNA序列并测序,进行同源性比对和系统进化分析.结果显示:从10份样本中分离到64株菌,从中筛选出产纤维素酶芽孢杆菌共23株,其中蜡样芽孢杆菌16株,苏云金芽孢杆菌7株.系统进化分析结果显示,蜡样芽孢杆菌共分为两大支,有一株菌单独聚为一支,另外15株菌聚为一支,其中这一支又分为五小支,具有一定的遗传多样性;7株苏云金芽孢杆菌分布在一个大支上.该结果为了解牦牛瘤胃产纤维素酶芽孢杆菌的种类和开发牦牛专用微生态制剂奠定了试验基础.  相似文献   

18.
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.  相似文献   

19.
旨在研究益生菌互作对肉鸡生长性能、肠道养分消化吸收及相关糖转运蛋白的影响。本研究中,200只1日龄雄性爱拔益加肉雏鸡被随机分为4组,每组5个重复,每个重复10只。将干酪乳杆菌、嗜酸乳杆菌和双歧杆菌分别按比例混合后于牛乳中发酵。对照组正常饮水(control组),益生菌Ⅰ组(混合比例为2:1:1)、Ⅱ组(混合比例为1:2:1)、Ⅲ组(混合比例为1:1:2)补充1%复合菌乳制品于饮用水中,各组皆饲喂基础日粮。整个试验饲养周期为42 d,每21 d为一个阶段。结果表明:1)与对照组相比,益生菌互作组均可显著提高肉鸡平均日增重(P<0.05),改善饲料利用率(P>0.05),在所有益生菌互作组中,益生菌Ⅰ组的肉鸡生长性能最好;2)益生菌互作显著刺激了空肠淀粉酶、脂肪酶、胰蛋白酶的特异性活性(P<0.05);3)益生菌互作组均显著提高了小肠绒毛高度(P<0.05),降低了隐窝深度(P<0.05),上调了GLUT2 mRNA的表达(P<0.05),其中,以益生菌Ⅰ、Ⅲ组作用最佳。综上所述,益生菌互作能够增强消化酶活性、改善消化道结构、上调营养转运蛋白表达,提高养分的消化率及能量吸收有效性,从而提高肉鸡的生产性能;且当干酪乳杆菌、嗜酸乳杆菌和双歧杆菌的混合配比为2:1:1时,复合益生菌制剂对肉鸡的作用效果较佳。  相似文献   

20.
The aim of this study was to investigate the effects of probiotic interaction on growth performance, intestinal nutrient digestion and absorption, and related sugar transporters in broiler. Two hundred 1-day-old male Arbor Acres broilers were selected, and then randomly divided into 4 groups with 5 replicates per group and 10 for each replicate. Lactobacillus casei, Lactobacillus acidophilus and Bifidobacterium lactis were mixed in proportion, and then fermented in milk. The chicks in the control group received normal drinking water, the chicks in probiotic group Ⅰ (mixed ratio of 2:1:1), group Ⅱ (mixed ratio of 1:2:1), and group III (mixed ratio of 1:1:2) received drinking water supplemented with 1% compound probiotics. All chickens were fed with normal diet for 42 days, every 21 days as a stage. The results showed that: 1) Compared to control group, the ADG(P<0.05) and feed conversion ratio (P>0.05) of broilers in all probiotic groups were improved. Furthermore, the production performance of broilers in probiotic group Ⅰ was higher than those of probiotic group Ⅱ and Ⅲ. 2) Compared to control group, specific activities of amylase, lipase, trypsin in jejunum of probiotic groups increased significantly (P<0.05). 3) In all probiotic groups, the intestinal villus height increased significantly (P<0.05), crypt depth decreased significantly (P<0.05), and the expression of glucose transporter 2 (GLUT2), particularly in probiotic group Ⅰ and Ⅲ, was significantly upregulated (P<0.05). In conclusion, probiotics interaction can enhance digestive enzyme activity, improve the structure of digestive tract, upregulate the expression of nutrient transporter, improve the nutrient digestibility and energy absorption efficiency, and further improve production performance of broiler. When the mixing ratio of L. casei, L. acidophilus and B. lactis was 2:1:1, the compound probiotics had a better effect on broilers.  相似文献   

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