吉林省猪致病性链球菌的分离与鉴定
摘要
猪链球菌病是分布范围极广常见的猪的传染病之一，危害严重，世界各国均有发生，也是多年来一直困扰我国养猪业的主要传染病之一。由于链球菌血清型较多，抗原结构复杂，在临床上又可呈现不同的症状，如败血症、脑膜脑炎、心内膜炎、肺炎、化脓性淋巴结炎、关节炎等，还常常与其他疾病发生合并感染。近几年来，该病的发病率和死亡率有逐年上升趋势。链球菌的致病性已引起临床工作者和科研人员的广泛关注和重视。本病在我国各省、市不同猪场均有发生，特别是在集约化养猪场中，其发生率有不断上升趋势，目前已成为我国猪场最常见的细菌性传染病之一。国外于1945年首次报道本病。国内是于1949年在上海首次发现本病，自20世纪60年代初期开始大面积流行，70年代末遍及全国大部分省、市。国外一些大猪场目前也受到本病严重扰乱。该病是我国法定二类动物疫病。据近几年的临床调查得知，我国的大小猪场均有发生，但主要集中在春秋两季的哺乳仔猪中，发病率为2％～5％，死亡率为40％左右。
目前，对我省猪链球菌病的发生规律、病原特性以及防治措施还不十分清楚，单纯靠药物又不能有效的控制该病的发生和蔓延。因此我们从病原学着手研究，以便为控制该病提供理论依据。2001～2002年我们对长春市、吉林市，延边市、四平等市、县较大的猪场所发生的急性、热性、败血性传染病进行了流行病学调查、临床诊断、病理检剖。在初步诊断为猪败血性链球菌病的基础上，从诸多病料中进行了病原分离，得到了20株猪源致病性链球菌。结合培养特性、生化特性及血清学反应等对其进行了分群鉴定。结果表明其形态、染色、培养特征及理化特性符合链球菌的特点，大多具有α或β溶血，生化试验结果不一致，根据检测结果初步认为：5株为兰氏分类法中的C群，9株为D群，3株为E群，1株为A群，2株未能定群。在病原性实验中15株链球菌对小鼠有性强弱不同致病，本试验可证实吉林省地方性猪链球菌病病原菌血清型已不局限于原有的C群链球菌。通过对15株病原性菌株的药敏性试验得知：大多数菌株对常用抗菌药有耐药性。
研制本地区的多价灭活苗是控制该病发生的根本方法。在吉林省地区性猪链球
吉林农业大学硕士学位论文
吉林省猪致病性链球菌的分离与鉴定
菌流矛洲朱检测的基础上，我们湘匕迄了有代表性的F6菌株进行了全面的生化特性鉴
定，为研制了多价灭活苗提供了参考依据。
目前，美国、英国、法国、俄罗斯、日本、印度、丹麦、荷兰、加拿大等国均
有关于猪链球菌分离鉴定及血清型分型的报道，从患败血症和脑炎的猪体内分离的
链球菌主要是R群的猪链球菌H型，和S群的猪链球菌I型，只有少数C群、D群
和L群引起心内膜炎和败血症的病原菌。这与我们的分离菌株的血清型是有一定差
异的。
Pig streptococcosis is a wide spread infectious disease which has occurred everywhere in the world and it also is one of the main infectious disease that puzzling pig farms in our country these years. Because of the great varity of streptothrix serum type and the complex structure of antigen, it presents different clinical symptom, such as septicemia, meningoencephalitis, endocarditis, pneumonia's, porodenia arthritis etc. It often occurs combinating with other disease. There is arising tendency of infection rate and death rate in recent year. Extensive attention is paid by clinicians and scientific researchers. It has occurred everywhere in our country, and the rate is in an rising trend especially in assembling pig farms. Now it is one of the most known bacterium disease. The first report of it aboard was in 1945, and 1949 in our country. It spreaded fast in 1960's, and in 1970's it has been reported in most area in our country. It is defined the second class animal epidemic disease by law. Based on the investigation in recent years, piglets in JiLin province are mainly occurred in spring and autumn. The infection rate is about 2% to 5%,and the death rate is about 40%.
Now we are not very clear with occurrence regulation and the original characteristic of pig streptococcosis disease . Its' infection and spread can not be effectively controlled simply by medicines. So we study it from pathogen.Between 2001 and 2002,we did epidemilogy survey, clinical diagnosis and pathological anatomy to acute, thermal and sepsis infectious disease in big pig farms in Chanchun, JiLin, Yanji and Gongzhuling. Based on the preliminary diagnosis of pig septic streptococcosis, pathogen separation was done, and 20 Pig Pathogenicity streptothrix were separated. Assortment identification was done through the combination of medium character, biochemistry character and serum reaction, its morphotype, chromosome, medium character and biochemistry character were in coincidence with pig streptococcosis. The result of fluidity test in biochemistry experiment showed that there were five strains belong to C group streptothrix, nine strains belong to D group streptothrix, three strains belong to E group streptothrix, one belong to A group and two strains can not be confined. In this experiment, there were fifteen strains streptothrix had different action in mice pathogenicity. This invistigation showed that Jilin
local pig streptococcosis serum type are not confined in C group streptothrix.
The development of pig streptothrix polyvalent inactivated vaccine is the pressing obligation to control this disease. Based on Jilin local pig streptothrix popular strains, we have chosen the representative F6 strain to be proceeded completely characteristic authenticates, which can provide consult data for research of manufacture of polyvalent inactivated vaccine.
At present, there are reports of pig streptothrix in America, England, France, Russia, Japan, India, Denmark, Canada etc. The streptothrix which separated from septic and cerebritis pig body mainly belong to type II of R group pig streptothrix and type I of S group pig streptothrix, only small amount of C group, D group and L group streptothrix cause endocarditis and septicema, which has some difference in according to our serum type.
目前，对我省猪链球菌病的发生规律、病原特性以及防治措施还不十分清楚，单纯靠药物又不能有效的控制该病的发生和蔓延。因此我们从病原学着手研究，以便为控制该病提供理论依据。2001～2002年我们对长春市、吉林市，延边市、四平等市、县较大的猪场所发生的急性、热性、败血性传染病进行了流行病学调查、临床诊断、病理检剖。在初步诊断为猪败血性链球菌病的基础上，从诸多病料中进行了病原分离，得到了20株猪源致病性链球菌。结合培养特性、生化特性及血清学反应等对其进行了分群鉴定。结果表明其形态、染色、培养特征及理化特性符合链球菌的特点，大多具有α或β溶血，生化试验结果不一致，根据检测结果初步认为：5株为兰氏分类法中的C群，9株为D群，3株为E群，1株为A群，2株未能定群。在病原性实验中15株链球菌对小鼠有性强弱不同致病，本试验可证实吉林省地方性猪链球菌病病原菌血清型已不局限于原有的C群链球菌。通过对15株病原性菌株的药敏性试验得知：大多数菌株对常用抗菌药有耐药性。
研制本地区的多价灭活苗是控制该病发生的根本方法。在吉林省地区性猪链球
吉林农业大学硕士学位论文
吉林省猪致病性链球菌的分离与鉴定
菌流矛洲朱检测的基础上，我们湘匕迄了有代表性的F6菌株进行了全面的生化特性鉴
定，为研制了多价灭活苗提供了参考依据。
目前，美国、英国、法国、俄罗斯、日本、印度、丹麦、荷兰、加拿大等国均
有关于猪链球菌分离鉴定及血清型分型的报道，从患败血症和脑炎的猪体内分离的
链球菌主要是R群的猪链球菌H型，和S群的猪链球菌I型，只有少数C群、D群
和L群引起心内膜炎和败血症的病原菌。这与我们的分离菌株的血清型是有一定差
异的。
Pig streptococcosis is a wide spread infectious disease which has occurred everywhere in the world and it also is one of the main infectious disease that puzzling pig farms in our country these years. Because of the great varity of streptothrix serum type and the complex structure of antigen, it presents different clinical symptom, such as septicemia, meningoencephalitis, endocarditis, pneumonia's, porodenia arthritis etc. It often occurs combinating with other disease. There is arising tendency of infection rate and death rate in recent year. Extensive attention is paid by clinicians and scientific researchers. It has occurred everywhere in our country, and the rate is in an rising trend especially in assembling pig farms. Now it is one of the most known bacterium disease. The first report of it aboard was in 1945, and 1949 in our country. It spreaded fast in 1960's, and in 1970's it has been reported in most area in our country. It is defined the second class animal epidemic disease by law. Based on the investigation in recent years, piglets in JiLin province are mainly occurred in spring and autumn. The infection rate is about 2% to 5%,and the death rate is about 40%.
Now we are not very clear with occurrence regulation and the original characteristic of pig streptococcosis disease . Its' infection and spread can not be effectively controlled simply by medicines. So we study it from pathogen.Between 2001 and 2002,we did epidemilogy survey, clinical diagnosis and pathological anatomy to acute, thermal and sepsis infectious disease in big pig farms in Chanchun, JiLin, Yanji and Gongzhuling. Based on the preliminary diagnosis of pig septic streptococcosis, pathogen separation was done, and 20 Pig Pathogenicity streptothrix were separated. Assortment identification was done through the combination of medium character, biochemistry character and serum reaction, its morphotype, chromosome, medium character and biochemistry character were in coincidence with pig streptococcosis. The result of fluidity test in biochemistry experiment showed that there were five strains belong to C group streptothrix, nine strains belong to D group streptothrix, three strains belong to E group streptothrix, one belong to A group and two strains can not be confined. In this experiment, there were fifteen strains streptothrix had different action in mice pathogenicity. This invistigation showed that Jilin
local pig streptococcosis serum type are not confined in C group streptothrix.
The development of pig streptothrix polyvalent inactivated vaccine is the pressing obligation to control this disease. Based on Jilin local pig streptothrix popular strains, we have chosen the representative F6 strain to be proceeded completely characteristic authenticates, which can provide consult data for research of manufacture of polyvalent inactivated vaccine.
At present, there are reports of pig streptothrix in America, England, France, Russia, Japan, India, Denmark, Canada etc. The streptothrix which separated from septic and cerebritis pig body mainly belong to type II of R group pig streptothrix and type I of S group pig streptothrix, only small amount of C group, D group and L group streptothrix cause endocarditis and septicema, which has some difference in according to our serum type.
引文
(1) 韩文瑜，何昭阳，刘玉斌：病原细菌检验技术，长春：吉林科学技术出版社，1992，7．
(2) 曹澍泽：兽医微生物学及免疫学技术，第一板，北京：北京农业大学出版社，1992，5．
(3) 莱曼：猪病学(刘文军，张中直等主校)，第六版，北京农业大学出版社，1990
(4) 王世若等，兽医微生物学及免疫学，237-241，中国人民解放军大学训练部出版，1983年9月第一版
(5) 甘肃农业大学主编，兽医微生物，173，农业出版社，1980年7月第一版．
(6) 汪诚天等，兽医实验检验手册，161，上海科技出版社，1984年6月第一板
(7) 张宝龙等，中国兽医杂志，1996，22(8)：37
(8) 宋阳威等，肥育猪急性败血性链球菌的诊治，畜牧与兽医，2002，34(1)：32—33
(9) 刁有祥等，猪链球菌病的诊治，中国兽医杂志，1999，5(25)，22
(10) 张大维等，猪败血性链球菌病的诊断和防治，河南畜牧兽医，2002，2(23)：29—30
(11) 蔡宝祥等主编，实用家畜传染病学，上海科技出版社，1989
(12) 中国农业科学院哈尔滨兽医研究所主编，家畜传染病学，北京，农业出版社1989
(13) 蔡宝祥等主编，动物传染病学，江苏技出版社，1993
(14) 李佑民主编，家畜传染病学，北京，蓝天出版社，1993
(15) 黑根等原著，胡祥璧等译，家畜传染病学，第七版，北京，农业出版社，1998
(16) Tarradas C, et al, journal of Clinical Microbiology, Feb,1994,578-580
[17] Higgins. R, et, al, Vet. Diagn. Invst, Microbiology, 1995,7:405-406
[18] Salasia. S.1.0. et, al, Veterinary Mirobiology, 1995,45:151-156
[19] Uri Vecht et al, Veterinary Microbiology,,1996,51:125-136
[20] Jacobs,A.A.C. et al, Veterinary Record, 1995,137:295-296
[21] 陆承平主编兽医微生物学 第三版 中国农业出版社2001．9
[22] 费恩阁主编 动物传染病学 第一版 吉林科技出版社1995．8
[23] 杨本升等主编 动物微生物学 第一版 吉林科技出版社1995．8
[24] 高云飞，苏亚君，李宝臣等 猪链球菌活疫苗的研制中国预防兽医学报2001，23：228-230
[25] 王白振，陈丽颖，杜君等 河南省猪链球菌病病原分群鉴定 河南农业科学2001，4：28-29
[26] 尹燕博，徐瑞，李葳等 猪链球菌病原和病理学诊断 中国动物检疫1997，14：3-5
[27] 孙银华，严勋，黄荣斌等 猪兰氏Q群链球菌病的诊断与防制 中国预防兽医学报1999，21：462-465
[28] 艾国良，陶钧 猪链球菌的诊断与防治畜牧兽医杂志1998，17：37-38
[29] 薛占永，杨旭，张会铭 规模化种猪场链球菌病的诊治2002，38：30-31
[30] 宋阳威，焦库华 育肥猪急性败血性链球菌病的诊治2002，34：32-33
(2) 曹澍泽：兽医微生物学及免疫学技术，第一板，北京：北京农业大学出版社，1992，5．
(3) 莱曼：猪病学(刘文军，张中直等主校)，第六版，北京农业大学出版社，1990
(4) 王世若等，兽医微生物学及免疫学，237-241，中国人民解放军大学训练部出版，1983年9月第一版
(5) 甘肃农业大学主编，兽医微生物，173，农业出版社，1980年7月第一版．
(6) 汪诚天等，兽医实验检验手册，161，上海科技出版社，1984年6月第一板
(7) 张宝龙等，中国兽医杂志，1996，22(8)：37
(8) 宋阳威等，肥育猪急性败血性链球菌的诊治，畜牧与兽医，2002，34(1)：32—33
(9) 刁有祥等，猪链球菌病的诊治，中国兽医杂志，1999，5(25)，22
(10) 张大维等，猪败血性链球菌病的诊断和防治，河南畜牧兽医，2002，2(23)：29—30
(11) 蔡宝祥等主编，实用家畜传染病学，上海科技出版社，1989
(12) 中国农业科学院哈尔滨兽医研究所主编，家畜传染病学，北京，农业出版社1989
(13) 蔡宝祥等主编，动物传染病学，江苏技出版社，1993
(14) 李佑民主编，家畜传染病学，北京，蓝天出版社，1993
(15) 黑根等原著，胡祥璧等译，家畜传染病学，第七版，北京，农业出版社，1998
(16) Tarradas C, et al, journal of Clinical Microbiology, Feb,1994,578-580
[17] Higgins. R, et, al, Vet. Diagn. Invst, Microbiology, 1995,7:405-406
[18] Salasia. S.1.0. et, al, Veterinary Mirobiology, 1995,45:151-156
[19] Uri Vecht et al, Veterinary Microbiology,,1996,51:125-136
[20] Jacobs,A.A.C. et al, Veterinary Record, 1995,137:295-296
[21] 陆承平主编兽医微生物学 第三版 中国农业出版社2001．9
[22] 费恩阁主编 动物传染病学 第一版 吉林科技出版社1995．8
[23] 杨本升等主编 动物微生物学 第一版 吉林科技出版社1995．8
[24] 高云飞，苏亚君，李宝臣等 猪链球菌活疫苗的研制中国预防兽医学报2001，23：228-230
[25] 王白振，陈丽颖，杜君等 河南省猪链球菌病病原分群鉴定 河南农业科学2001，4：28-29
[26] 尹燕博，徐瑞，李葳等 猪链球菌病原和病理学诊断 中国动物检疫1997，14：3-5
[27] 孙银华，严勋，黄荣斌等 猪兰氏Q群链球菌病的诊断与防制 中国预防兽医学报1999，21：462-465
[28] 艾国良，陶钧 猪链球菌的诊断与防治畜牧兽医杂志1998，17：37-38
[29] 薛占永，杨旭，张会铭 规模化种猪场链球菌病的诊治2002，38：30-31
[30] 宋阳威，焦库华 育肥猪急性败血性链球菌病的诊治2002，34：32-33