犬源乳酸菌的分离鉴定及生化特性研究
|
摘要
当今因滥用抗生素，而出现的畜产品药物残留及耐药菌株增加等问题十分突出。
益生素(Probiotics)的出现为解决以上问题提供了新思路。
本次试验首次从犬的直肠内容物及粪便中分离并筛选了2株细菌。这2株细菌
经过细菌的菌落形态、菌体形态及生化特性鉴定出了其所属的种属，分别为棒状乳
酸杆菌、瑞士乳酸杆菌。
分离的乳酸菌菌株经过耐酸试验、胆盐耐受试验、对犬肠粘膜上皮细胞结合性
试验、滤菌培养液抑菌试验及急性毒性试验检验其性质。分离的乳酸菌菌株可抵抗
动物胃肠道环境中不利于细菌生长的因素，从而可较少损失的达到肠道。分离乳酸
菌菌株对犬肠粘膜上皮细胞有比较强的结合性，可与之稳固的粘合，从而能稳定的
存在于犬的胃肠道中，以防止因肠蠕动而排出体外。该乳酸菌菌株对50只昆明小白
鼠的急性毒性试验表明，该乳酸菌菌株对动物机体无急性毒性作用。而经滤过除菌
分离乳酸菌培养液的抑菌试验表明，该乳酸菌的分泌物对葡萄球菌、大肠杆菌有一
定的抑制作用。
将乳酸菌的培养液应用于哺乳仔犬，对试验组仔犬、对照组仔犬比较后发现试
验组仔犬平均日增重比对照组仔犬提高14.76％，而腹泻率下降6.22％。
由此可见分离的乳酸菌菌株安全无毒。乳酸菌的培养液经滤过除菌后对葡萄球
菌、大肠杆菌有一定的抑制作用，从而能有效地抑制胃肠道内有害菌群的生长，该
乳酸菌的培养液饲喂给哺乳仔犬取得了明显的增重作用及预防腹泻效果。分离的乳
酸菌菌株可作为犬的益生素微生态制剂的备选菌株。
Nowadays, the problems were obviously stood out such as the drug remaining, the drug resistant bacteria increasing ,etc, because of abusing antibiotic. The appearance of probiotics provided new idea for solving these problems.
Two strains of bacteria were isolated from the rectal contents of 30 canines in this experiment. The two stains of bacteria were identified the species and genus which they belong to by their colony forms , thallus forms and biological properties , and the two strains of bacteria were Rod Lactobacillus , Swizerland Lactobacillus, respectively.
The isolated Lactobacillus strains were passed the tests of tolerance to acid and bile salt, adherence to the epithelial cells of the canine intestinal mucous membrane, the inhibiting germ of filtering germs culture media and acute toxicity .It suggested that the isolated Lactobacillus strains can resist to the agents which were not fit for the growth of the germs in the animal intestinal environment, the above tests. And the isolated Lactobacillus could strongly adherence to the epithelial cells of canine intestinal mucous membrane. So the Lactobacillus can combine with the cells steadily so that the Lactobacillus could exist in the canine gastrointestinal tract, which prevent to be expelled from the body because of vermiculation of intestine .It suggested that the isolated Lactobacillus strains were no acute toxicity to the animal body from the acute toxicity test of the isolated Lactobacillus strains to 50 small Kunming white mice. And it suggested that the secretion of the isolated Lactobacillus strains
could inhibit the familiar pathogenical germs from the inhibition germ test of the culture media of isolated Lactobacillus which removed the germs by filtered.
The culture media of isolated Lactobacillus which were tested no toxicity to the animal body by acute toxicity test was fed to the young suckling canine .It can be found that the daily weight gain of the young controlled canine group was higher than that of the young compared canine group (it increased 14.76%), while the diarrhea rate of the young
controlled canine group was lower than that of the young compared group (it descended 6.22%).
Above all, the isolated Lactobacillus strains are safe and no toxicity to the animal, can get to the canine gastrointestinal tract in losing very few , adhere to the epithelial cells of canine intestinal mucous membrane , which proved that the isolated Lactobacillus strains could steadily in the intestinal tract . The filtered removing germs culture media could inhibit the familiar pathogenical germs, so the isolated Lactobacillus could effectively inhibit the growth of harmful flora in the gastrointestinal tract. The obvious effects of weight gain and preventing diarrhea were obtained by the cultured media of isolated Lactobacillus feeding to the young suckling canine.
In conclusion, the isolated Lactobacillus strains could be used as the micro-ecosystem preparation of probiotic for the canine.
益生素(Probiotics)的出现为解决以上问题提供了新思路。
本次试验首次从犬的直肠内容物及粪便中分离并筛选了2株细菌。这2株细菌
经过细菌的菌落形态、菌体形态及生化特性鉴定出了其所属的种属，分别为棒状乳
酸杆菌、瑞士乳酸杆菌。
分离的乳酸菌菌株经过耐酸试验、胆盐耐受试验、对犬肠粘膜上皮细胞结合性
试验、滤菌培养液抑菌试验及急性毒性试验检验其性质。分离的乳酸菌菌株可抵抗
动物胃肠道环境中不利于细菌生长的因素，从而可较少损失的达到肠道。分离乳酸
菌菌株对犬肠粘膜上皮细胞有比较强的结合性，可与之稳固的粘合，从而能稳定的
存在于犬的胃肠道中，以防止因肠蠕动而排出体外。该乳酸菌菌株对50只昆明小白
鼠的急性毒性试验表明，该乳酸菌菌株对动物机体无急性毒性作用。而经滤过除菌
分离乳酸菌培养液的抑菌试验表明，该乳酸菌的分泌物对葡萄球菌、大肠杆菌有一
定的抑制作用。
将乳酸菌的培养液应用于哺乳仔犬，对试验组仔犬、对照组仔犬比较后发现试
验组仔犬平均日增重比对照组仔犬提高14.76％，而腹泻率下降6.22％。
由此可见分离的乳酸菌菌株安全无毒。乳酸菌的培养液经滤过除菌后对葡萄球
菌、大肠杆菌有一定的抑制作用，从而能有效地抑制胃肠道内有害菌群的生长，该
乳酸菌的培养液饲喂给哺乳仔犬取得了明显的增重作用及预防腹泻效果。分离的乳
酸菌菌株可作为犬的益生素微生态制剂的备选菌株。
Nowadays, the problems were obviously stood out such as the drug remaining, the drug resistant bacteria increasing ,etc, because of abusing antibiotic. The appearance of probiotics provided new idea for solving these problems.
Two strains of bacteria were isolated from the rectal contents of 30 canines in this experiment. The two stains of bacteria were identified the species and genus which they belong to by their colony forms , thallus forms and biological properties , and the two strains of bacteria were Rod Lactobacillus , Swizerland Lactobacillus, respectively.
The isolated Lactobacillus strains were passed the tests of tolerance to acid and bile salt, adherence to the epithelial cells of the canine intestinal mucous membrane, the inhibiting germ of filtering germs culture media and acute toxicity .It suggested that the isolated Lactobacillus strains can resist to the agents which were not fit for the growth of the germs in the animal intestinal environment, the above tests. And the isolated Lactobacillus could strongly adherence to the epithelial cells of canine intestinal mucous membrane. So the Lactobacillus can combine with the cells steadily so that the Lactobacillus could exist in the canine gastrointestinal tract, which prevent to be expelled from the body because of vermiculation of intestine .It suggested that the isolated Lactobacillus strains were no acute toxicity to the animal body from the acute toxicity test of the isolated Lactobacillus strains to 50 small Kunming white mice. And it suggested that the secretion of the isolated Lactobacillus strains
could inhibit the familiar pathogenical germs from the inhibition germ test of the culture media of isolated Lactobacillus which removed the germs by filtered.
The culture media of isolated Lactobacillus which were tested no toxicity to the animal body by acute toxicity test was fed to the young suckling canine .It can be found that the daily weight gain of the young controlled canine group was higher than that of the young compared canine group (it increased 14.76%), while the diarrhea rate of the young
controlled canine group was lower than that of the young compared group (it descended 6.22%).
Above all, the isolated Lactobacillus strains are safe and no toxicity to the animal, can get to the canine gastrointestinal tract in losing very few , adhere to the epithelial cells of canine intestinal mucous membrane , which proved that the isolated Lactobacillus strains could steadily in the intestinal tract . The filtered removing germs culture media could inhibit the familiar pathogenical germs, so the isolated Lactobacillus could effectively inhibit the growth of harmful flora in the gastrointestinal tract. The obvious effects of weight gain and preventing diarrhea were obtained by the cultured media of isolated Lactobacillus feeding to the young suckling canine.
In conclusion, the isolated Lactobacillus strains could be used as the micro-ecosystem preparation of probiotic for the canine.
引文
[1] 张振兴．我国养殖业生物安全的现状与对策[J]．畜牧兽医科技信息．2002，18(4)：3～5．
[2] 臧建军．猪禽肠道微生物调控的研究[J]．中国饲料．2002，16：10～11．
[3] 程伶．益生素的作用及在生产中的应用[J]．国外畜牧科技．1992，19(5)：38～39．
[4] 贾士芳，郭兴华．活菌制剂的现状和未来[J]．生物工程进展．1996，16(2)：16～21．
[5] Peter H.A Sneath. Bergy's Manual of Systematic Bacteriology, Williams&Wilkins 1986.
[6] 杨建军．乳酸菌及其在畜牧生产中的应用[J]．畜禽业．2002，18(8)：36～37．
[7] 王欣，顾均华．饲用微生态制剂[J]．中国饲料．2002，11：13～14．
[8] 帅丽芳．微生态制剂对反刍动物消化系统的调控作用[J]．中国饲料．2002，9：16～17．
[9] Hosono A et al.Agric.Biol.Chem., 1990,45,1639.
[10] G Perdigon et al.J.Dairy Res.1991, 58,485.
[11] R K Chandra et al.J.Dairy Res.1991, 58,485.
[12] 石军，孙德文，陈安国．益生素在水产养殖中的应用研究进展[J]．中国饲料．2002，14：29～31．
[13] 谢正昀主编．微生物培养基的制造与应用[M]．中国农业出版社，1995．
[14] 罗珍兰．乳酸菌胶囊粪肠球菌的分离鉴定及生长特性观察[J]．江苏农学院学报．1996，17(4)：91～94．
[15] 廖延雄主编．兽医微生物试验诊断手册[M]．中国农业出版社，1995．
[16] 王海荣．肉仔鸡嗉囊中乳酸菌的分离与鉴定[J]．内蒙古农业大学学报．2000，2：19～23．
[17] 方祥．乳酸菌的分离、鉴定及其生长特性研究[J]．中国微生态杂志．2000，5：262～264．
[18] 顾瑞霞．胆汁酸盐和低pH值对乳酸菌活性的影响[J]．微生物学报．1996，3：144～146．
[19] 赵瑞香．嗜酸杆菌在模拟胃肠环境中抗性的研究[J]．微生物学通报．2002，29(2)：35～38．
[20] Annika Mayra-Makinen, Merja Manninen, Gyllenberg H. The adherence of lactic acid bacteria to the columnar epithelial cells of pigs and calves .Journal of Applied Bacteriology, 1983, 55:241～245.
[21] 马向前，周德庆．双歧杆菌和乳酸菌的一种简便快速记数法[J]．微生物学报．1997，1：62～64．
[22] 王世荣．乳酸菌代谢产物对常见致病菌的体外抑菌试验[J]．中国微生态学杂志．1990，2(2)：38．
[23] 张均田主编．现代药理试验方法[M]，北京医科大学中国协和医科大学联合出版社1998．
[24] 王传彬，王永坤，朱国强等．动物源乳杆菌筛选及生物特性参数测定[J]．江苏农学院学报．1987，18(1)：1～5
[25] 禹慧明．益生乳杆菌的筛选及特性研究[J]．微生物学通报．2002，29(1)：53-56．
[26] 赵京杨，吕茂洲，张卫宏等．加酶益生素添加时间对哺乳断乳仔猪日增重和腹泻率的影响[J]．中国畜牧杂志．2002，38(3)：19～20．
[27] 刘萌武．云南省乳品中乳杆菌的鉴定[J]．微生物学报．1993，4：203～206．
[28] 王琦．内蒙古地区乳汁品中乳酸菌的分离、鉴定[J]．内蒙古农牧学院学报．1997，2：1～6．
[29] 乌尼．内蒙古锡林郭勒盟两个旗乳与乳制品中乳杆菌的分离与鉴定[J]．内蒙古农牧学院学报．1998，2：1～10．
[30] 叶子坚．乳酸菌微胶囊化的研究[J]．微生物学报．2000，5：507～512．
[31] 李少英．内蒙古双峰驼乳及乳制品中乳杆菌的分离及其生物学特性的研究[J]．内蒙古农业大学学报．2001，2：80～85．
[32] 敖敦格日勒．阿鲁科尔沁草原牛乳中乳酸菌生物学的研究[J]．内蒙古农业大学学报．2001，3：17～22．
[33] 周雨霞．内蒙古部分地区乳及乳制品中乳链球菌的分离及鉴定[J]．内蒙古农牧大学学报．1995，3：55～59．
[34] 郝永清．呼和浩特市及召河等地牛乳微生物生态学初探[J]．内蒙古农牧大学学报．1992，3：7～11．
[35] 傅月华．动物肠道内乳酸菌的分离和应用[J]．饲料研究．1994，5：11～12．
[36] 孔建．益生素生产菌乳链球菌SB900的分离及生物特性研究[J]．微生物学报．1995，6：450～454．
[37] 袁清珠．都尔伯特半荒漠草原牧区绵山羊乳及乳制品中乳杆菌的生物学特性的研究[J]．内蒙古农业大学学报．2001，3：23～27．
[38] 白贵蓉．贺兰山和腾格里沙漠牧区山羊乳中乳酸球菌的分离及其生物学特性研究[J]．内蒙古农牧大学学报．2000，3：39～45．
[39] 杨瑞鹏．几种酸泡菜中乳酸菌的分离与鉴定[J]．华中农业大学学报．1987，4：346～350．
[40] 孟和毕力格．内蒙古双峰驼乳及乳制品中乳杆菌的分离及其生物学特性的研究[J]．内蒙古农业大学学报．2001，22(2)：80～85．
[41] 李清等．乳酸杆菌的培养与分离[J]．中国饲料．2002，5：12～13．
[42] 程立忠，沙涛．人体嗜酸乳杆菌的分离与筛选[J]．云南农业大学(自然科学版)．1998，20(2)：108～111．
[43] 东秀珠编著．常见细菌系统鉴定手册[M]，科学出版社，2001．
[44] M.T．马迪根．微生物生物学[M]，科学出版社，2001．
[45] Hood S K, Zottola E A. Effect of low pH on the ability of lactobacillus acidophilus to survive and adhere to human intestinal cells.J Food Sci,1988,53(5): 1514～1516.
[46] Gonway P L. Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells. J Dairy Sci, 1987,70:1～12.
[47] 康白主编．微生态学[M]，大连出版社，1988．
[48] 胥清富．复合活菌剂对肉用仔鸡生产性能、消化和盲肠菌群的影响[J]．中国畜牧杂志．1999，35(5)：12～13．
[49] 马桂荣．SL益生素的免疫促进作用闭．中华微生物学和免疫学杂志．1994，14(1)：32．
[50] 胡东量．乳酸菌的抗肿瘤、抗变异原及免疫增强作用[J]．中国乳品工业．1997，25(6)：11～14．
[51] 何清明．猪微生物饲料添加剂(8701)对生长肥育猪抗病、增重及提高饲料利用率研究[J]．中国微生态学杂志．1999，2(4)：43～47．
[52] 程惠平．三种微生态制剂治疗急性腹泻的费用—效果分析[J]．中国微生态学杂志．2002，14(1)：38～39．
[53] Gukor G, et al. Human viral gastroenteritis. Microbiol Rev 1984;48(2) :157.
[54] Gill C. The push toward probiotics .Feeding International 1986; 11:8.
[55] Maxweel CV, et al .Effect of probiotic supplementation on performance ,Fecal parameters and digestibility in growing finishing swine. J Anim Sci 1982;55(suppl.):284.
[56] Pollmann DS. Probiofics in pig diets. Recent Advances in Animal. Nutrition 1986; 2:30.
[57] 丁玉华．益生素“活清”对肉仔鸡生产性能的影响[J]．中国饲料．2002，9：11～12．
[58] Naveh M W. Adherence pili in avian strains of E.coli: Effect on pathogencity [J]. Avian Diseases, 1984,28(3): 651-661.
[59] 潘宝海．益生菌在养鸡生产中的应用[J]．中国饲料．2000，16：9～11．
[60] 夏兆刚．直接饲喂微生物在畜牧生产上的应用[J]．中国饲料．2000，9：28～29．
[61] 李芳益．益菌多在治疗和预防仔猪腹泻上的应用[J]．中国饲料．2002，19：13．
[62] 袁莉民．双歧杆菌在仔猪肠道的结合特性研究[J]．江苏农业研究．1999，20(4)：61～64．
[63] 康白．对回春生生产菌种DM8504菌株的研究报告[J]，中国微生态学杂志．1994，6(1)：1～6．
[64] Marie F B,Dominque B,Jean R N, et al .Adhension of human bifidobacterial strains to cultured human intestinal epithelical cells and inhibition of enterpathogen-cell interations [J]. Applied and Environmental Microbiology ,1992, 59(12):4121-4128.
[65] 胡文锋．热灭活嗜酸乳杆菌及其多肽抗菌物质特性研究[J]．饲料博览．1999，11(10)：7～8．
[66] 胡文锋．遭遇康长乐托肠健宝抑制或杀灭动物肠道致病菌的机理及特点[J]．中国饲料，1999，9：17～19．
[67] 蒋文泓．一种新型药物和饲料添加剂—动物微生态制剂[J]．兽药与饲料添加剂．2000，5(1)：18～20．
[68] GILAND S ,T E STALEV and BUSH LJ. Importance of Bile Tolerance of Lactobacillus acidophilus Used as a Dietary Adjunct [J]. Journal of Dairy Science, 1984,(12):3045-3050.
[69] Fuller R. Probiotics 2: Applications and Practical Aspects, Chapman & Hall Ltd, London ,UK, 1997.
[2] 臧建军．猪禽肠道微生物调控的研究[J]．中国饲料．2002，16：10～11．
[3] 程伶．益生素的作用及在生产中的应用[J]．国外畜牧科技．1992，19(5)：38～39．
[4] 贾士芳，郭兴华．活菌制剂的现状和未来[J]．生物工程进展．1996，16(2)：16～21．
[5] Peter H.A Sneath. Bergy's Manual of Systematic Bacteriology, Williams&Wilkins 1986.
[6] 杨建军．乳酸菌及其在畜牧生产中的应用[J]．畜禽业．2002，18(8)：36～37．
[7] 王欣，顾均华．饲用微生态制剂[J]．中国饲料．2002，11：13～14．
[8] 帅丽芳．微生态制剂对反刍动物消化系统的调控作用[J]．中国饲料．2002，9：16～17．
[9] Hosono A et al.Agric.Biol.Chem., 1990,45,1639.
[10] G Perdigon et al.J.Dairy Res.1991, 58,485.
[11] R K Chandra et al.J.Dairy Res.1991, 58,485.
[12] 石军，孙德文，陈安国．益生素在水产养殖中的应用研究进展[J]．中国饲料．2002，14：29～31．
[13] 谢正昀主编．微生物培养基的制造与应用[M]．中国农业出版社，1995．
[14] 罗珍兰．乳酸菌胶囊粪肠球菌的分离鉴定及生长特性观察[J]．江苏农学院学报．1996，17(4)：91～94．
[15] 廖延雄主编．兽医微生物试验诊断手册[M]．中国农业出版社，1995．
[16] 王海荣．肉仔鸡嗉囊中乳酸菌的分离与鉴定[J]．内蒙古农业大学学报．2000，2：19～23．
[17] 方祥．乳酸菌的分离、鉴定及其生长特性研究[J]．中国微生态杂志．2000，5：262～264．
[18] 顾瑞霞．胆汁酸盐和低pH值对乳酸菌活性的影响[J]．微生物学报．1996，3：144～146．
[19] 赵瑞香．嗜酸杆菌在模拟胃肠环境中抗性的研究[J]．微生物学通报．2002，29(2)：35～38．
[20] Annika Mayra-Makinen, Merja Manninen, Gyllenberg H. The adherence of lactic acid bacteria to the columnar epithelial cells of pigs and calves .Journal of Applied Bacteriology, 1983, 55:241～245.
[21] 马向前，周德庆．双歧杆菌和乳酸菌的一种简便快速记数法[J]．微生物学报．1997，1：62～64．
[22] 王世荣．乳酸菌代谢产物对常见致病菌的体外抑菌试验[J]．中国微生态学杂志．1990，2(2)：38．
[23] 张均田主编．现代药理试验方法[M]，北京医科大学中国协和医科大学联合出版社1998．
[24] 王传彬，王永坤，朱国强等．动物源乳杆菌筛选及生物特性参数测定[J]．江苏农学院学报．1987，18(1)：1～5
[25] 禹慧明．益生乳杆菌的筛选及特性研究[J]．微生物学通报．2002，29(1)：53-56．
[26] 赵京杨，吕茂洲，张卫宏等．加酶益生素添加时间对哺乳断乳仔猪日增重和腹泻率的影响[J]．中国畜牧杂志．2002，38(3)：19～20．
[27] 刘萌武．云南省乳品中乳杆菌的鉴定[J]．微生物学报．1993，4：203～206．
[28] 王琦．内蒙古地区乳汁品中乳酸菌的分离、鉴定[J]．内蒙古农牧学院学报．1997，2：1～6．
[29] 乌尼．内蒙古锡林郭勒盟两个旗乳与乳制品中乳杆菌的分离与鉴定[J]．内蒙古农牧学院学报．1998，2：1～10．
[30] 叶子坚．乳酸菌微胶囊化的研究[J]．微生物学报．2000，5：507～512．
[31] 李少英．内蒙古双峰驼乳及乳制品中乳杆菌的分离及其生物学特性的研究[J]．内蒙古农业大学学报．2001，2：80～85．
[32] 敖敦格日勒．阿鲁科尔沁草原牛乳中乳酸菌生物学的研究[J]．内蒙古农业大学学报．2001，3：17～22．
[33] 周雨霞．内蒙古部分地区乳及乳制品中乳链球菌的分离及鉴定[J]．内蒙古农牧大学学报．1995，3：55～59．
[34] 郝永清．呼和浩特市及召河等地牛乳微生物生态学初探[J]．内蒙古农牧大学学报．1992，3：7～11．
[35] 傅月华．动物肠道内乳酸菌的分离和应用[J]．饲料研究．1994，5：11～12．
[36] 孔建．益生素生产菌乳链球菌SB900的分离及生物特性研究[J]．微生物学报．1995，6：450～454．
[37] 袁清珠．都尔伯特半荒漠草原牧区绵山羊乳及乳制品中乳杆菌的生物学特性的研究[J]．内蒙古农业大学学报．2001，3：23～27．
[38] 白贵蓉．贺兰山和腾格里沙漠牧区山羊乳中乳酸球菌的分离及其生物学特性研究[J]．内蒙古农牧大学学报．2000，3：39～45．
[39] 杨瑞鹏．几种酸泡菜中乳酸菌的分离与鉴定[J]．华中农业大学学报．1987，4：346～350．
[40] 孟和毕力格．内蒙古双峰驼乳及乳制品中乳杆菌的分离及其生物学特性的研究[J]．内蒙古农业大学学报．2001，22(2)：80～85．
[41] 李清等．乳酸杆菌的培养与分离[J]．中国饲料．2002，5：12～13．
[42] 程立忠，沙涛．人体嗜酸乳杆菌的分离与筛选[J]．云南农业大学(自然科学版)．1998，20(2)：108～111．
[43] 东秀珠编著．常见细菌系统鉴定手册[M]，科学出版社，2001．
[44] M.T．马迪根．微生物生物学[M]，科学出版社，2001．
[45] Hood S K, Zottola E A. Effect of low pH on the ability of lactobacillus acidophilus to survive and adhere to human intestinal cells.J Food Sci,1988,53(5): 1514～1516.
[46] Gonway P L. Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells. J Dairy Sci, 1987,70:1～12.
[47] 康白主编．微生态学[M]，大连出版社，1988．
[48] 胥清富．复合活菌剂对肉用仔鸡生产性能、消化和盲肠菌群的影响[J]．中国畜牧杂志．1999，35(5)：12～13．
[49] 马桂荣．SL益生素的免疫促进作用闭．中华微生物学和免疫学杂志．1994，14(1)：32．
[50] 胡东量．乳酸菌的抗肿瘤、抗变异原及免疫增强作用[J]．中国乳品工业．1997，25(6)：11～14．
[51] 何清明．猪微生物饲料添加剂(8701)对生长肥育猪抗病、增重及提高饲料利用率研究[J]．中国微生态学杂志．1999，2(4)：43～47．
[52] 程惠平．三种微生态制剂治疗急性腹泻的费用—效果分析[J]．中国微生态学杂志．2002，14(1)：38～39．
[53] Gukor G, et al. Human viral gastroenteritis. Microbiol Rev 1984;48(2) :157.
[54] Gill C. The push toward probiotics .Feeding International 1986; 11:8.
[55] Maxweel CV, et al .Effect of probiotic supplementation on performance ,Fecal parameters and digestibility in growing finishing swine. J Anim Sci 1982;55(suppl.):284.
[56] Pollmann DS. Probiofics in pig diets. Recent Advances in Animal. Nutrition 1986; 2:30.
[57] 丁玉华．益生素“活清”对肉仔鸡生产性能的影响[J]．中国饲料．2002，9：11～12．
[58] Naveh M W. Adherence pili in avian strains of E.coli: Effect on pathogencity [J]. Avian Diseases, 1984,28(3): 651-661.
[59] 潘宝海．益生菌在养鸡生产中的应用[J]．中国饲料．2000，16：9～11．
[60] 夏兆刚．直接饲喂微生物在畜牧生产上的应用[J]．中国饲料．2000，9：28～29．
[61] 李芳益．益菌多在治疗和预防仔猪腹泻上的应用[J]．中国饲料．2002，19：13．
[62] 袁莉民．双歧杆菌在仔猪肠道的结合特性研究[J]．江苏农业研究．1999，20(4)：61～64．
[63] 康白．对回春生生产菌种DM8504菌株的研究报告[J]，中国微生态学杂志．1994，6(1)：1～6．
[64] Marie F B,Dominque B,Jean R N, et al .Adhension of human bifidobacterial strains to cultured human intestinal epithelical cells and inhibition of enterpathogen-cell interations [J]. Applied and Environmental Microbiology ,1992, 59(12):4121-4128.
[65] 胡文锋．热灭活嗜酸乳杆菌及其多肽抗菌物质特性研究[J]．饲料博览．1999，11(10)：7～8．
[66] 胡文锋．遭遇康长乐托肠健宝抑制或杀灭动物肠道致病菌的机理及特点[J]．中国饲料，1999，9：17～19．
[67] 蒋文泓．一种新型药物和饲料添加剂—动物微生态制剂[J]．兽药与饲料添加剂．2000，5(1)：18～20．
[68] GILAND S ,T E STALEV and BUSH LJ. Importance of Bile Tolerance of Lactobacillus acidophilus Used as a Dietary Adjunct [J]. Journal of Dairy Science, 1984,(12):3045-3050.
[69] Fuller R. Probiotics 2: Applications and Practical Aspects, Chapman & Hall Ltd, London ,UK, 1997.