枯草芽孢杆菌对黄曲霉毒素B_(1)引起羊人工瘤胃异常发酵的改善作用  

Improvement Effects of Bacillus subtilis on Abnormal Fermentation in Artificial Rumen Induced by Aflatoxin B_(1) in Sheep

作　　者：张照杰 韩一铭 高宇凤 任晓丽 宋超 李志娟 李洪政 石冬梅[2,3] 贾雨婷 姬中豪 皇甫和平 王金明[1] ZHANG Zhaojie;HAN Yiming;GAO Yufeng;REN Xiaoli;SONG Chao;LI Zhijuan;LI Hongzheng;SHI Dongmei;JIA Yuting;JI Zhonghao;HUANGFU Heping;WANG Jinming(College of Veterinary Medicine,Shanxi Agricultural University,Taigu 030801,China;College of Veterinary Medicine,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China;Zhengzhou Key Laboratory of Animal Nutritional and Metabolic Diseases and Poisoning Diseases,Zhengzhou 450046,China;Shangqiu Meilan Bio-Engineering Co.,Ltd.,Shangqiu 476299,China;Henan Guokang Detection Technology Co.,Ltd.,Zhengzhou 450001,China)

机构地区：[1]山西农业大学动物医学学院,太谷030801 [2]河南牧业经济学院动物医药学院,郑州450046 [3]郑州市动物营养代谢病与中毒病重点实验室,郑州450046 [4]商丘美兰生物工程有限公司,商丘476299 [5]河南国康检测技术有限公司,郑州450001

出　　处：《动物营养学报》2025年第2期1260-1270,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：新一轮河南省重点学科(学科名称:兽医,学科序号:312);河南省科技攻关项目(232102111037)。

摘　　要：本试验旨在探究添加不同剂量枯草芽孢杆菌(Bacillus subtilis)对黄曲霉毒素B_(1)(AFB_(1))致羊人工瘤胃异常发酵的影响。试验以全混合日粮(TMR)为发酵底物,采用单因素试验设计,分为7个组,包括对照组(NC组)、AFB_(1)组和5个试验组,其中NC组添加1 mL甲醇,AFB_(1)组添加1 mL 10μg/mL的AFB_(1),试验组分别同时添加1 mL 10μg/mL的AFB_(1)和0.01%、0.03%、0.06%、0.12%、0.24%的枯草芽孢杆菌制剂,每组3个重复。试验当天随机收集3只屠宰羊的新鲜瘤胃液,按照试验设计在39℃条件下培养4、8、12、24和48 h,记录pH和产气量,测定氨态氮(NH_(3)-N)、微生物蛋白(MCP)和挥发性脂肪酸(VFA)含量,并利用荧光定量PCR检测微生物区系。结果表明:1)发酵24 h时,AFB_(1)组产气量与NC组相比显著降低(P<0.05),0.03%组产气量与AFB_(1)组相比极显著升高(P<0.01)。2)发酵24 h时,各组发酵液pH均无显著差异(P>0.05)。AFB_(1)组NH_(3)-N和MCP含量低于NC组,但无显著差异(P>0.05);0.03%组NH_(3)-N和MCP含量与AFB_(1)组相比极显著升高(P<0.01)。3)发酵24 h时,AFB_(1)组发酵液各VFA含量在数值上均低于NC组,其中异丁酸含量与NC组相比显著降低(P<0.05);0.03%组乙酸、丁酸和总挥发性脂肪酸(TVFA)与AFB_(1)组相比极显著提高(P<0.01),丙酸和异丁酸含量与AFB_(1)组相比显著提高(P<0.05)。4)与NC组相比,AFB_(1)组产琥珀酸丝状杆菌(Fibrobacter succinogenes)、嗜淀粉瘤胃杆菌(Ruminobacter amylophilus)和原虫相对丰度极显著降低(P<0.01),溶纤维丁酸弧菌(Butyrivibrio fibrisolvens)和栖瘤胃普雷沃氏菌(Prevotella ruminicola)相对丰度显著降低(P<0.05)。与AFB_(1)组相比,0.03%组白色瘤胃球菌(Ruminococcus albus)、黄色瘤胃球菌(Ruminococcus flavefaciens)、栖瘤胃普雷沃氏菌和产甲烷菌相对丰度显著提高(P<0.05),溶纤维丁酸弧菌、嗜淀粉瘤胃杆菌和原虫相对丰度极显著提高(P<0.01)。综上所述,AFB_(1)会降低体外发酵瘤胃产气量�This experiment was conducted to investigate the effects of different doses of Bacillus subtilis supplementation on abnormal fermentation in artificial rumen induced by aflatoxin B_(1)(AFB_(1))in sheep.Using a single factor experimental design and total mixed ration(TMR)as fermentation substrate,the experiment was divided into 7 groups,including control group(NC group),AFB_(1) group and 5 experimental groups.The NC group was supplemented with 1 mL methanol,the AFB_(1) group was supplemented with 1 mL 10μg/mL AFB_(1),and the experimental groups were supplemented with 1 mL 10μg/mL AFB_(1) and Bacillus subtilis preparations of 0.01%,0.03%,0.06%,0.12% and 0.24%,respectively,with 3 replicates in each group.On the day of the experiment,the fresh rumen fluid from 3 slaughtered sheep was randomly collected and cultured at 39℃ for 4,8,12,24 and 48 h according to the experimental design.pH and gas production were recorded,the contents of ammonia nitrogen(NH_(3)-N),microbial protein(MCP)and volatile fatty acid(VFA)were determined,and the microbiota was detected by fluorescence quantitative PCR.The results showed as follows:1)At 24 h of fermentation,the gas production in AFB_(1) group was significantly decreased compared with NC group(P<0.05),and the gas production in 0.03% group was extremely significantly increased compared with AFB_(1) group(P<0.01).2)At 24 h of fermentation,there was no significant difference in pH in fermentation broth among all groups(P>0.05).The contents of NH_(3)-N and MCP in AFB_(1) group were lower than those in NC group,but there was no significant difference(P>0.05);the contents of NH_(3)-N and MCP in 0.03% group were extremely significantly higher than those in AFB_(1) group(P<0.01).3)At 24 h of fermentation,the each VFA content in fermentation broth in AFB_(1) group was numerically lower than that in NC group,and the isobutyrate content was significantly lower than that in NC group(P<0.05);the contents of acetate,butyrate and total volatile fatty acid(TVFA)in 0.03% group were extremely sign

关 键 词：枯草芽孢杆菌 黄曲霉毒素B_(1) 瘤胃 体外发酵 微生物区系 

分 类 号：S816[农业科学—饲料科学]