上海地区猪链球菌病流行情况调查及风险控制分析与实践
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摘要
猪链球菌病(Swine Streptococcosis)是链球菌属((Streptococcus)中马链球菌兽疫亚种(S. equi ssp. zooepidemicus)、马链球菌类马亚种(S. equi ssp. equisimilis)、猪链球菌(S. suis)、类猪链球菌(S. porcinus)、停乳链球菌类马亚种(S. dysgalactiae ssp. equisimilis)等链球菌引致猪疫病的总称。临床上主要表现为淋巴结脓肿、脑膜炎、关节炎以及败血症为主要特征,其中以败血症的危害最大,在某些特定诱因作用下,发病猪群的死亡率可以达到80%。其中,猪链球菌是世界范围内引致猪链球菌病最主要的病原,该茵可引起猪脑膜炎以及败血症等疫病,人通过特定的传播途径亦可感染该菌。近年来猪链球茵病在我国广泛流行,特别是猪链球菌2型、马链球菌兽疫亚种感染,严重影响着我国的养殖业,造成了很大的经济损失。
通过对1998～2010年13年临床病例的回顾调查及猪场的采样监测,了解猪链球菌在畜间、空间分布的情况、以及茵株毒力和耐药性的变异情况、为防控本病提供依据。
通过对1998～2010年上海及江苏、浙江、福建、江西、安徽等周边地区的猪链球菌病病例回顾调查和菌株收集,并于2009～2010年发放并回收《猪链球菌病调查表》,开展血清学、病原学监测等手段开展调查研究。结果表明,调查地区猪群链球菌病征复杂多样,败血症型和关节炎型最多,分别占30.13%和26.99%,其次为脑膜脑炎型和心内膜炎型分别占22.8%和14.64%,脑膜脑炎型近年来有上升趋势,临床上多与猪蓝耳病病毒、猪圆环病毒、伪狂犬病毒、副猪嗜血杆菌、大肠杆菌、沙门茵等形成多重感染,对宿主造成更大的危害。
实验证实,猪链球菌是目前临床病例的优势菌株(47.78%),其中猪链球菌2型有109株,占21.08%,其他分离到猪链球菌还包括1、3、4、5、7、8、9、l0、11、13、15、16、25、28等血清型。C群链球菌仍占有一定比例(16.05%),其中马链球菌兽疫亚种(SeZ)有48株,占9.28%。另外排除猪链球菌交叉反应后D群有41株(7.93%)、B群有15株(2.9%)、A群1株(0.19%),值得注意的是有159株(30.75%)不属于A～D、F和G群,也不是猪链球菌。说明猪链球菌2型作为一种危害严重的人畜共患病病原,已经成为主要的致病菌。健康猪群猪链球菌带茵率为8.55%,分离到2、3、4、5、9、10、11、13、15、16、19、21、22、25、26、29等血清型猪链球菌,其中15型最多(13.24%),其次为2型(10.5%)、29型(6.85%)、26型(5.48%)、3型(5.48%),另外从3家猪场的猪舍空气样本中检出24株猪链球菌,分离率为1.45%,其中3型4株,15型1株,29型2株,17株未鉴定血清型。
采用7种毒力因子两组多重PCR方法,检测63株猪链球菌临床分离株及26株健康猪群分离株,结果发现,高毒力基因型mrp+epf+sly+gdh+gapdh+orf2+fbps+的菌株在上海地区猪链球菌2型分离菌株中占主导地位(68.75%),健康猪携带菌株也具有临床菌株相似的毒力因子(60.86%)。106株猪源链球菌药敏试验结果表明,分离菌株已对多种抗生素产生了耐药性,对林可胺类、四环素类、大环内酯类、氨基糖苷类药物高度耐药,对磺胺类药物产生40%左右的耐药性；以往临床首选的青霉素、氨苄青霉素,也分别有34.9%和11.3%的菌株产生了耐药性。
为控制猪场链球菌感染的风险,降低猪链球菌发病率。对猪链球菌感染的可能风险因素进行了分析,引入HACCP理念,绘制了猪场猪链球菌感染的流程图,识别出猪场猪链球菌感染的风险因子及关键控制点(CCP-like):隐性带菌猪、混群饲养、精液、病毒性免疫抑制病(猪蓝耳病、PCV-2感染、猪伪狂犬病等)、疫苗免疫、药物预防。针对各关键控制点制定出相应的防控措施,为猪链球菌的防控提供技术思路。
为能控制猪场链球茵感染的风险,根据识别出的猪链球菌感染的风险因子和关键控制点,制定了一套适用于上海地区猪场的猪链球菌综合防控措施。控制重点为隐性带菌及病死猪、猪链球菌疫苗免疫结合药物防治、二点四段猪群流动管理、猪蓝耳病、PCV-2感染、猪伪狂犬病等免疫抑制病的控制等。控制措施在嘉定区、浦东新区、松江区、崇明县年出栏为1万头以上的4个猪场以及2个农户示范应用,并定期进行监测和验证。结果表明,示范猪场猪链球菌隐性带茵猪的的比例由实施前的9.12%(31头)下降至6.47%(22头),下降了2-3%,猪链球菌2型带菌率由实施前的1.47%(5头),下降至0.29%(1头),效果均显著(p<0.05),但该措施的实施对猪场管理、设施等要求较高,需在良好操作规范的基础上实施,否则效果并不明显,如管理不到位的示范猪场四。
Swine Streptococcosis is a variety of pig disease can caused by Streptococcus equi ssp. zooepidemicus, S. equi ssp. equisimilis, S. suis, S. porcinus, S. dysgalactiae ssp. equisimilis and other Streptococcus. The major clinical manifestations include septicemia, endocarditis, meningitis, arthritis, lymph node abscess, and the septicemia, of which the most damaging, pigs mortality up to80%under certain incentives action. S. suis type2is the most prevalent type in association with diseases in most countries. It is also an important zoonotic agent for humans in contact with diseased pigs or their products, causing life threatening diseases.
Through review of clinical cases from1998to2010, sampling surveys and monitoring of pig farms, we have mastered Streptococcus suis distribution in livestock and space, as well as virulence and drug resistance variation, and Provided the basis for the prevention and control of the disease.
After the systemic epidemiological investigation in Shanghai, Jiangsu, Zhejiang, Fujian, Jiangxi, Anhui and other surrounding areas from1998to2010, we found that247(47.78%) were identified as S.suis strains during the517Streptococcus clinical isolates from1998to2010and it had substituted S. equi subsp. zooepidemicus prevailing in China. During the clinical S.suis isolates, the serotype2was most frequent(44.13%) which followed by the serotype9(7.69%),7(3.24%),5(3.24%),3(3.24%),1(2.83%),4(2.83%),25(2.02%). During the healthy pigs isolates, the serotypel5was most frequent(13.24%) which followed by the serotype2(10.5%),29(6.85%),26(5.48%) and3(5.48%. In addition,24(1.45%) S. suis isolates were detected in the air samples from three pig farms, include serotype3(4),29(2),15(1).
To investigate the distribution of virulence factors of S. suis isolated from clinical case and healthy pigs, seven virulence genes were detected by PCR, including the glutamate dehydrogenase gene (gdh), the suilysin gene (sly), the extracellular factor gene (epf), the muramidase-released protein gene (mrp), the fibronectin-binding protein gene (fpbs), the glyceraldehyde-3-phosphate dehydrogenase (gadph) gene and the virulence-associated sequence orf2. The results showed the distribution of virulent genes of S.suis in Shanghai was complicated, with12genetypes detected among71serotype2isolates.68.75%of the serotype2clinical isolates and60.76%of healthy isolates were with the genotype of mrp+epf+sly+gdh+gapdh+orf2+fbps+, which were well known to be highly virulent.106clinical isolates of Streptococcus susceptibility test results show that the isolates have produced a variety of antibiotic resistance, highly resistant to Lim amines, tetracyclines, macrolides, aminoglycosides, and40%of the strains have produced Sulfa drugs resistance.34.9%and11.3%of the strains have produced penicillin, ampicillin drug resistance respectively.
S. suis infection, an emerging zoonosis, is an increasing public health problem across China and South East Asia. Little is known of the risk factors underlying the disease. To control the risk of S. suis infection, the potential risk factors of S. suis infection were analyzed and a process control system known as "hazard analysis critical control point,"(HACCP) were introduced. Through the epidemiological assessment of this disease, this control study has identified the important risk factors associated with S. suis infection. After Potential risk factors were assessed using HACCP procedure, the Critical control point (CCP-like) were identified, that is carrier pigs, mixed flocks, semen, immunosuppressive disease (PRRS, PCV-2, PR, etc.), vaccine、 drug prevention. For each critical control point to develop appropriate prevention and control measures for prevention and control of S. suis infection technical ideas.
In order to control the risk of S. suis infection, according to identified risk factors and critical control points, a comprehensive prevention and control measures were developed for S. suis control in Shanghai. The main control objects and measures were the carrier, vaccine combined with drug prevention, two poin and four segment pigs flow management, PRRS, PCV-2, PR and other immunosuppressive diseases control. The control measures were demonstrated applications and monitored in four farms and two farmers in the Jiading District, Pudong District,, Songjiang District, Chongming County. The results show that the S. suis carrier in the demonstration farms from9.12%(31) decreased to6.47%(22), a decrease of2to3%,and the carrier rate of S. suis type2from1.47%(5) dropped to0.29%(1), the effect was significantly (p<0.05). However, the effect is not obvious in the demonstration farms4.
通过对1998～2010年13年临床病例的回顾调查及猪场的采样监测,了解猪链球菌在畜间、空间分布的情况、以及茵株毒力和耐药性的变异情况、为防控本病提供依据。
通过对1998～2010年上海及江苏、浙江、福建、江西、安徽等周边地区的猪链球菌病病例回顾调查和菌株收集,并于2009～2010年发放并回收《猪链球菌病调查表》,开展血清学、病原学监测等手段开展调查研究。结果表明,调查地区猪群链球菌病征复杂多样,败血症型和关节炎型最多,分别占30.13%和26.99%,其次为脑膜脑炎型和心内膜炎型分别占22.8%和14.64%,脑膜脑炎型近年来有上升趋势,临床上多与猪蓝耳病病毒、猪圆环病毒、伪狂犬病毒、副猪嗜血杆菌、大肠杆菌、沙门茵等形成多重感染,对宿主造成更大的危害。
实验证实,猪链球菌是目前临床病例的优势菌株(47.78%),其中猪链球菌2型有109株,占21.08%,其他分离到猪链球菌还包括1、3、4、5、7、8、9、l0、11、13、15、16、25、28等血清型。C群链球菌仍占有一定比例(16.05%),其中马链球菌兽疫亚种(SeZ)有48株,占9.28%。另外排除猪链球菌交叉反应后D群有41株(7.93%)、B群有15株(2.9%)、A群1株(0.19%),值得注意的是有159株(30.75%)不属于A～D、F和G群,也不是猪链球菌。说明猪链球菌2型作为一种危害严重的人畜共患病病原,已经成为主要的致病菌。健康猪群猪链球菌带茵率为8.55%,分离到2、3、4、5、9、10、11、13、15、16、19、21、22、25、26、29等血清型猪链球菌,其中15型最多(13.24%),其次为2型(10.5%)、29型(6.85%)、26型(5.48%)、3型(5.48%),另外从3家猪场的猪舍空气样本中检出24株猪链球菌,分离率为1.45%,其中3型4株,15型1株,29型2株,17株未鉴定血清型。
采用7种毒力因子两组多重PCR方法,检测63株猪链球菌临床分离株及26株健康猪群分离株,结果发现,高毒力基因型mrp+epf+sly+gdh+gapdh+orf2+fbps+的菌株在上海地区猪链球菌2型分离菌株中占主导地位(68.75%),健康猪携带菌株也具有临床菌株相似的毒力因子(60.86%)。106株猪源链球菌药敏试验结果表明,分离菌株已对多种抗生素产生了耐药性,对林可胺类、四环素类、大环内酯类、氨基糖苷类药物高度耐药,对磺胺类药物产生40%左右的耐药性；以往临床首选的青霉素、氨苄青霉素,也分别有34.9%和11.3%的菌株产生了耐药性。
为控制猪场链球菌感染的风险,降低猪链球菌发病率。对猪链球菌感染的可能风险因素进行了分析,引入HACCP理念,绘制了猪场猪链球菌感染的流程图,识别出猪场猪链球菌感染的风险因子及关键控制点(CCP-like):隐性带菌猪、混群饲养、精液、病毒性免疫抑制病(猪蓝耳病、PCV-2感染、猪伪狂犬病等)、疫苗免疫、药物预防。针对各关键控制点制定出相应的防控措施,为猪链球菌的防控提供技术思路。
为能控制猪场链球茵感染的风险,根据识别出的猪链球菌感染的风险因子和关键控制点,制定了一套适用于上海地区猪场的猪链球菌综合防控措施。控制重点为隐性带菌及病死猪、猪链球菌疫苗免疫结合药物防治、二点四段猪群流动管理、猪蓝耳病、PCV-2感染、猪伪狂犬病等免疫抑制病的控制等。控制措施在嘉定区、浦东新区、松江区、崇明县年出栏为1万头以上的4个猪场以及2个农户示范应用,并定期进行监测和验证。结果表明,示范猪场猪链球菌隐性带茵猪的的比例由实施前的9.12%(31头)下降至6.47%(22头),下降了2-3%,猪链球菌2型带菌率由实施前的1.47%(5头),下降至0.29%(1头),效果均显著(p<0.05),但该措施的实施对猪场管理、设施等要求较高,需在良好操作规范的基础上实施,否则效果并不明显,如管理不到位的示范猪场四。
Swine Streptococcosis is a variety of pig disease can caused by Streptococcus equi ssp. zooepidemicus, S. equi ssp. equisimilis, S. suis, S. porcinus, S. dysgalactiae ssp. equisimilis and other Streptococcus. The major clinical manifestations include septicemia, endocarditis, meningitis, arthritis, lymph node abscess, and the septicemia, of which the most damaging, pigs mortality up to80%under certain incentives action. S. suis type2is the most prevalent type in association with diseases in most countries. It is also an important zoonotic agent for humans in contact with diseased pigs or their products, causing life threatening diseases.
Through review of clinical cases from1998to2010, sampling surveys and monitoring of pig farms, we have mastered Streptococcus suis distribution in livestock and space, as well as virulence and drug resistance variation, and Provided the basis for the prevention and control of the disease.
After the systemic epidemiological investigation in Shanghai, Jiangsu, Zhejiang, Fujian, Jiangxi, Anhui and other surrounding areas from1998to2010, we found that247(47.78%) were identified as S.suis strains during the517Streptococcus clinical isolates from1998to2010and it had substituted S. equi subsp. zooepidemicus prevailing in China. During the clinical S.suis isolates, the serotype2was most frequent(44.13%) which followed by the serotype9(7.69%),7(3.24%),5(3.24%),3(3.24%),1(2.83%),4(2.83%),25(2.02%). During the healthy pigs isolates, the serotypel5was most frequent(13.24%) which followed by the serotype2(10.5%),29(6.85%),26(5.48%) and3(5.48%. In addition,24(1.45%) S. suis isolates were detected in the air samples from three pig farms, include serotype3(4),29(2),15(1).
To investigate the distribution of virulence factors of S. suis isolated from clinical case and healthy pigs, seven virulence genes were detected by PCR, including the glutamate dehydrogenase gene (gdh), the suilysin gene (sly), the extracellular factor gene (epf), the muramidase-released protein gene (mrp), the fibronectin-binding protein gene (fpbs), the glyceraldehyde-3-phosphate dehydrogenase (gadph) gene and the virulence-associated sequence orf2. The results showed the distribution of virulent genes of S.suis in Shanghai was complicated, with12genetypes detected among71serotype2isolates.68.75%of the serotype2clinical isolates and60.76%of healthy isolates were with the genotype of mrp+epf+sly+gdh+gapdh+orf2+fbps+, which were well known to be highly virulent.106clinical isolates of Streptococcus susceptibility test results show that the isolates have produced a variety of antibiotic resistance, highly resistant to Lim amines, tetracyclines, macrolides, aminoglycosides, and40%of the strains have produced Sulfa drugs resistance.34.9%and11.3%of the strains have produced penicillin, ampicillin drug resistance respectively.
S. suis infection, an emerging zoonosis, is an increasing public health problem across China and South East Asia. Little is known of the risk factors underlying the disease. To control the risk of S. suis infection, the potential risk factors of S. suis infection were analyzed and a process control system known as "hazard analysis critical control point,"(HACCP) were introduced. Through the epidemiological assessment of this disease, this control study has identified the important risk factors associated with S. suis infection. After Potential risk factors were assessed using HACCP procedure, the Critical control point (CCP-like) were identified, that is carrier pigs, mixed flocks, semen, immunosuppressive disease (PRRS, PCV-2, PR, etc.), vaccine、 drug prevention. For each critical control point to develop appropriate prevention and control measures for prevention and control of S. suis infection technical ideas.
In order to control the risk of S. suis infection, according to identified risk factors and critical control points, a comprehensive prevention and control measures were developed for S. suis control in Shanghai. The main control objects and measures were the carrier, vaccine combined with drug prevention, two poin and four segment pigs flow management, PRRS, PCV-2, PR and other immunosuppressive diseases control. The control measures were demonstrated applications and monitored in four farms and two farmers in the Jiading District, Pudong District,, Songjiang District, Chongming County. The results show that the S. suis carrier in the demonstration farms from9.12%(31) decreased to6.47%(22), a decrease of2to3%,and the carrier rate of S. suis type2from1.47%(5) dropped to0.29%(1), the effect was significantly (p<0.05). However, the effect is not obvious in the demonstration farms4.
引文
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[4]GB/T 17823-2009集约化猪场防疫基本要求
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[9]GB 16548-2006病害动物和病害动物产品生物安全处理规程
[10]NY 5027-2008无公害食品畜禽饮用水水质
[ll]GB 13078-2001饲料卫生标准
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