青海与河南部分地区猪旋毛虫病流行病学调查及虫种鉴定
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摘要
旋毛虫病(Trichinellosis)是一种呈世界性分布的重要食源性人兽共患寄生虫病,主要因生食或半生食含有旋毛虫(Trichinella)感染性幼虫的肉类及其制品所致。在过去的30年间,许多国家和地区旋毛虫病的暴发流行又出现了上升的趋势,已将其列入重新出现的疾病(re-emerging disease)。在我国,仅2004～2009年就报道了15次人体旋毛虫病暴发,其中12次是因生食或半生食猪肉所致。旋毛虫病不仅严重危害人体健康,对畜牧养殖业,尤其是养猪业也可造成巨大的经济损失。
目前已将旋毛虫属分为8个种和4个分类地位尚未确定的基因型。不同种或基因型旋毛虫的地理分布、宿主范围、冰冻耐力、感染性与致病性等生物学特征均存在差异,因而,旋毛虫属分类研究对旋毛虫病的病原学、临床治疗、流行病学及预防控制等均具有重要价值。旋毛虫的宿主范围甚广,国外已在野生动物体内发现8种旋毛虫,但我国目前只有关于旋毛虫(Trichinella spiralis, T1)和乡土旋毛虫(Trichinella nativa, T2)的报道,尚没有发现我国存在旋毛虫其它种或基因型,我国是否存在旋毛虫的其他种或基因型有待进一步调查。
本研究对我国青海和河南两省的部分地区进行了猪旋毛虫流行病学调查,将调查所获得的旋毛虫分离株接种到昆明小鼠体内进行保种,使用多重PCR方法对调查获取的旋毛虫分离株进行分子生物学鉴定和分类研究,进一步研究我国是否存在旋毛虫的其他种或基因型。本研究涉及旋毛虫病流行病学和分子生物学分类研究,可为旋毛虫病的致病机制提供更加科学、合理的病原生物学解释,更好的揭示其致病与地理分布及不同宿主间的差别与联系,为我国旋毛虫不同地理株的生物学分类研究和虫种鉴定提供基线资料和理论依据。
材料与方法
1.现场流行病学调查分别在我国青海和河南两省随机选取数个县市作为流行病学调查现场,选取待宰生猪做为调查对象。采样时通过动物疾病控制中心(ADC)或疾病预防控制中心(CDC)联系定点屠宰场负责人或检疫员,到集中屠宰点或农户自家宰杀生猪时进行肉样的采集,采样时从每头生猪的膈肌脚采集约50g肉样。
2.肉样的检测将采集到的猪肉样本逐一编号,编号后先使用压片镜检法对肉样进行初检。在镜检后对所有肉样均按照国际旋毛虫病委员会(ICT)推荐使用的人工消化法复检。
3.实验动物的接种雄性清洁级昆明小鼠,购自河南省实验动物中心,在本教研室动物饲养室按照实验动物管理条例进行饲养管理。将采集自同一调查地点的旋毛虫分离株的肌幼虫使用人工消化法进行分离,并采用灌胃法接种到已禁食1d的清洁级空白昆明小鼠体内。
4.旋毛虫肌幼虫的收集将接种不同旋毛虫分离株或参考株40d后的昆明小鼠引颈处死,确定旋毛虫接种成功后,将保种小鼠肌肉置搅拌器中搅碎,使用人工消化法对旋毛虫肌幼虫进行分离。收集到的旋毛虫幼虫直接用于提取基因组DNA或用灭菌生理盐水洗涤数次后冻存备用。
5.基因组DNA的提取及纯度检测使用酚氯仿法提取旋毛虫肌幼虫基因组DNA,使用U-2001型紫外可见分光光度计分别测A260和A280的OD值,并计算两者的比值(OD A260/A280),若两者比值介于1.8-1.9之间,表示提取的基因组DNA纯度较高。
6.多重PCR反应及产物鉴定本实验所用多重PCR引物采用Zarlenga(1999)设计的引物,反应体系及条件经过合理优化调整,使用2%琼脂糖凝胶对多重PCR扩增产物进行电泳,电泳结束后使用Gene Genius凝胶图象分析仪观察电泳结果,并对电泳结果进行鉴定分析。
1.青海省部分地区商品猪旋毛虫感染情况在位于青海省海东地区的互助土族自治县和海西地区首府德令哈市的多个采样点共对192头商品猪进行了采样,使用压片镜检法和人工消化法对膈肌样本进行了检疫,两个地区的猪肉均未检出旋毛虫幼虫。
2.河南省部分地区商品猪旋毛虫感染情况在河南省的安阳、商丘两地采样3个批次,检疫屠宰生猪共278头,使用压片镜检法进行检验时均未发现阳性肉样,而使用人工消化法复检时则检出18份阳性肉样,平均感染率为6.47%(18/278)；18份阳性肉样均来自本地农户圈养生猪,本地农户圈养生猪的旋毛虫感染率为10.84%(18/166),工业化集中养殖场的商品猪的旋毛虫感染率为0(0/112),不同饲养方式的生猪旋毛虫感染率之间的差异具有显著统计学意义(χ2=11.67,P<0.01)。本地农户圈养生猪的旋毛虫感染率,安阳为10.00%(9/90),商丘为11.84%(9/76),二者之间的差异不具有统计学意义(χ2=0.12,P>0.05)。
3.商品猪旋毛虫感染的强度使用人工消化法检疫18份阳性肉样时,共消化阳性肌肉样品122.8g,检出旋毛虫肌幼虫51条,阳性肉样的平均每克肌肉虫荷(1pg)为0.42(51/122.8),感染度较低。其中安阳的9份阳性肉样中平均每克肌肉虫荷为0.34(19/55.9),最低值为0.11,最高值为1.41；商丘的9份阳性肉样中均每克肌肉虫荷平均水平为0.48(32/66.9),最小值为0.10,最高值为1.58；二者的每克肌肉虫荷的最高值均超过了旋毛虫对公共卫生构成威胁的最低感染水平(1pg≥1)。对来自不同地方的阳性肉样的每克肌肉虫荷进行统计学分析,发现分别采集自安阳和商丘的阳性肉样虫荷之间的差异不具有统计学意义(Z=一0.05,P>0.05)。
4.多重PCR结果经过多重PCR扩增,我国河南安阳猪源旋毛虫分离株扩增出了一条大小为173bp的条带,该条带为旋毛虫(T1,ISS3)在多重PCR鉴定时的特有带型。河南安阳猪源旋毛虫分离株的电泳结果和河南南阳猪源地理株及旋毛虫(T1,ISS3)的结果一致,而不同于乡土旋毛虫(T2,127bp)、布氏旋毛虫(T3,127bp和253bp)、伪旋毛虫(T4,310bp)和纳氏旋毛虫(T7,155bp和404bp)。
结论
1.青海部分地区的屠宰猪未检出旋毛虫感染；河南省部分农村地区屠宰生猪的旋毛虫自然感染率较高,但感染强度较低。
2.河南省安阳地区猪源旋毛虫分离株属于旋毛形线虫(Trichinella spiralis, T1)。
Trichinellosis is an important food-borne parasitosis and parasitic zoonosis all over the world. When raw or undercooked meat or its products which contain infective Trichinella larvae were ingested, trichinellosis should occur. In recent years, outbreaks of human trichinellosis have taken place in many countries, and trichinellosis was defined as a re-emerging disease for its epidemiological trends. From 2004 to 2009, at least 15 outbreaks of trichinellosis were reported in China. Pork was the predominant source of outbreaks of human trichinellosis in China,12 out of 15 outbreaks were caused by eating raw or undercooked pork. Trichinellosis can cause expensive economic loss, not only harms to public health, but also to farming, especially the industrialized hoggery.
Eight species and 4 uncertain genotypes were recognised in the genus Trichinella till now. Because there were distinct differences among different species of Trichinella, for instance, morphology, geography distribution, the range of host, the resistance to freezing, the infecting ability to host and immunity response of host to different species, so the study on taxonomy of Trichinella is necessary. The aim of the study on Trichinella taxonomy is to offer valuable informations on the pathogen biology, the clinical treatment, the epidemiology, the prevention and control of trichinellosis in humans and animals. The range of Trichinella's hosts was very wide, and 8 species of Trichinella have been separated from wild animals. In China, only Trichinella spiralis (T1) and Trichinella nativa (T2) were reported till today, were there other species or genotypes of Trichinella exist in China still not sure.
In order to find the new Trichinella isolates in China, epidemiological investigation was carried on to separate novel isolates from Qinghai and Henan province in this study. The male Kunming mice were inoculated with new Trichinella isolates for species keeping. Further more, these different isolates of Trichinella were identified by multiplex PCR method. To provide the evidence for the relationship among pathogenesis, distribution and hosts, the epidemiology and molecular biology research were carried into execution in this study.
Materials and Methods
1. Epidemiological investigation The prevalence rates of swine trichinellosis in Qinghai and Henan Province were observed by epidemiological investigation. The sampling points were slected by local Animal Disease Control Center (ADC) and Center for Disease Control and Prevention (CDC). Samples were taken from diaphragm muscle and 50g each.
2. Samples inspection All the samples were numbered first, and quarantined by trichinelloscopy, then, all of them should be inspected again by artifical digestion method which recommended by International Commission on Trichinellosis (ICT).
3. Artificial inoculation of experimental animals Two month-old clean Kunming mice (18g～20g) were obtained from the Experimental Animal Center of Medical College of Zhengzhou University. After a whole day fast, male Kunming mice were orally inoculated with new Trichinella isolate from the same sampling point for species keeping.
4. Recovery of Trichinella muscle larvae The Kunming mice which infected with different isolates and international reference strains of Trichinella were killed 40 days later, and purified muscle larvae were collected by artifical digestion method.
5. Genomic DNA acquired and purity checked Genomic DNA was prepared according to phenyl-Chloroform method from muscle larvae. The purity of genomic DNA was checked by spectrophotometer (U-2001). OD value of A260 and A280 were detected by spectrophotometer first, and then the ratio of A260 to A280 was calculated for pure assessment. The purity of genomic DNA was fine when the ratio between 1.8-1.9.
6. Identification by multiplex PCR The genomic DNA which acquired from new isolate and different Trichinella international reference strains were identified by the method multiplex PCR. When the gene was amplied,5 pairs of primers that designed by Zarlenga (1999) were used. The products of multiplex PCR were electrophoresed in 2% agarose gel and visualised under Gene Genius Imaging System.
Results
1. The investigation on swine trichinellosis in Qinghai province In this investigation,192 pieces of meat was collected from diaphragm muscle in Huzu and Delingha of Qinghai province. Samples were quarantined by trichinelloscopy and digestion method, and none trichinella was found. All the samples were negative, none of the Trichinella infective swine was found in parts of Qinghai province.
2. The investigation on swine trichinellosis in Henan province In Henan province, sampling points were choosen from Anyang and Shangqiu city. Three groups of diaphragm muscle samples were collected from 278 swine. None Trichinella was found when quarantined by trichinelloscopy, but 18 positive samples were detected when the digestion method was used, the positive rate of all the samples was 6.47%(18/278). All the positive samples were collected from the swine that breeded in pig bed by local farmer, and the Trichinella infection rate of this kind of swine was 10.84%(18/166). The swine which from industrialized hoggery were free of Trichinella infection (0/112), and the differentiation was statistically significant (χ2=11.67, P<0.01) between local pig bed and industrialized hoggery of Trichinella infection rates. There was no difference of Trichinella infection rates in local pig bed between Anyang and Shangqiu (χ2=0.12,P>0.05).
3. Infective intensity of Trichinella in swine When 18 positive samples were detected by artifical digestion method, about 51 larvae were found from 122.8g of pork, and the larvae per gram (lpg) is 0.42. For the 9 positive samples that from Anyang county, the lpg has a range from 0.11 to 1.41, and the average level of lpg is 0.34 (19/55.9). Among the 9 Trichinella infective samples which from Sui county, the minimum of lpg is 0.10, the maximum is 1.58, with 0.48 (32/66.9) as its average. Both maxima of lpg are higher than the lowest level, which threaten to the public health (1pg≥1). The difference of lpg between Anyang county and Sui county without statistically significant (Z=-0.05, P>0.05).
4. Multiplex PCR Anyang isolate of Trichinella was identified by multiplex PCR method, and the result was observed by using electrophoresis. The Nanyang isolate and 5 Trichinella international reference strains were also analyzed as control at the same time. The special band (173bp) that belongs to Trichinella spiralis (T1, ISS3) was also showed by Nanyang and Anyang isolates, which was different from Trichinella nativa (T2,127bp), Trichinella britovi (T3,127bp and 253bp), Trichinella pesudospiralis (T4,310bp) and Trichinella nelsoni (T7,155bp and 404bp).
Conclusions
1. The swine infected with Trichinella are not found in part areas of Qinghai province. Though prevalence of swine trichinellosis is relatively high in rural part of Henan province, the intensity of infection is low.
2. The Trichinella isolates recovered from pork in Anyang were identified as Trichinella spiralis (T1).
目前已将旋毛虫属分为8个种和4个分类地位尚未确定的基因型。不同种或基因型旋毛虫的地理分布、宿主范围、冰冻耐力、感染性与致病性等生物学特征均存在差异,因而,旋毛虫属分类研究对旋毛虫病的病原学、临床治疗、流行病学及预防控制等均具有重要价值。旋毛虫的宿主范围甚广,国外已在野生动物体内发现8种旋毛虫,但我国目前只有关于旋毛虫(Trichinella spiralis, T1)和乡土旋毛虫(Trichinella nativa, T2)的报道,尚没有发现我国存在旋毛虫其它种或基因型,我国是否存在旋毛虫的其他种或基因型有待进一步调查。
本研究对我国青海和河南两省的部分地区进行了猪旋毛虫流行病学调查,将调查所获得的旋毛虫分离株接种到昆明小鼠体内进行保种,使用多重PCR方法对调查获取的旋毛虫分离株进行分子生物学鉴定和分类研究,进一步研究我国是否存在旋毛虫的其他种或基因型。本研究涉及旋毛虫病流行病学和分子生物学分类研究,可为旋毛虫病的致病机制提供更加科学、合理的病原生物学解释,更好的揭示其致病与地理分布及不同宿主间的差别与联系,为我国旋毛虫不同地理株的生物学分类研究和虫种鉴定提供基线资料和理论依据。
材料与方法
1.现场流行病学调查分别在我国青海和河南两省随机选取数个县市作为流行病学调查现场,选取待宰生猪做为调查对象。采样时通过动物疾病控制中心(ADC)或疾病预防控制中心(CDC)联系定点屠宰场负责人或检疫员,到集中屠宰点或农户自家宰杀生猪时进行肉样的采集,采样时从每头生猪的膈肌脚采集约50g肉样。
2.肉样的检测将采集到的猪肉样本逐一编号,编号后先使用压片镜检法对肉样进行初检。在镜检后对所有肉样均按照国际旋毛虫病委员会(ICT)推荐使用的人工消化法复检。
3.实验动物的接种雄性清洁级昆明小鼠,购自河南省实验动物中心,在本教研室动物饲养室按照实验动物管理条例进行饲养管理。将采集自同一调查地点的旋毛虫分离株的肌幼虫使用人工消化法进行分离,并采用灌胃法接种到已禁食1d的清洁级空白昆明小鼠体内。
4.旋毛虫肌幼虫的收集将接种不同旋毛虫分离株或参考株40d后的昆明小鼠引颈处死,确定旋毛虫接种成功后,将保种小鼠肌肉置搅拌器中搅碎,使用人工消化法对旋毛虫肌幼虫进行分离。收集到的旋毛虫幼虫直接用于提取基因组DNA或用灭菌生理盐水洗涤数次后冻存备用。
5.基因组DNA的提取及纯度检测使用酚氯仿法提取旋毛虫肌幼虫基因组DNA,使用U-2001型紫外可见分光光度计分别测A260和A280的OD值,并计算两者的比值(OD A260/A280),若两者比值介于1.8-1.9之间,表示提取的基因组DNA纯度较高。
6.多重PCR反应及产物鉴定本实验所用多重PCR引物采用Zarlenga(1999)设计的引物,反应体系及条件经过合理优化调整,使用2%琼脂糖凝胶对多重PCR扩增产物进行电泳,电泳结束后使用Gene Genius凝胶图象分析仪观察电泳结果,并对电泳结果进行鉴定分析。
1.青海省部分地区商品猪旋毛虫感染情况在位于青海省海东地区的互助土族自治县和海西地区首府德令哈市的多个采样点共对192头商品猪进行了采样,使用压片镜检法和人工消化法对膈肌样本进行了检疫,两个地区的猪肉均未检出旋毛虫幼虫。
2.河南省部分地区商品猪旋毛虫感染情况在河南省的安阳、商丘两地采样3个批次,检疫屠宰生猪共278头,使用压片镜检法进行检验时均未发现阳性肉样,而使用人工消化法复检时则检出18份阳性肉样,平均感染率为6.47%(18/278)；18份阳性肉样均来自本地农户圈养生猪,本地农户圈养生猪的旋毛虫感染率为10.84%(18/166),工业化集中养殖场的商品猪的旋毛虫感染率为0(0/112),不同饲养方式的生猪旋毛虫感染率之间的差异具有显著统计学意义(χ2=11.67,P<0.01)。本地农户圈养生猪的旋毛虫感染率,安阳为10.00%(9/90),商丘为11.84%(9/76),二者之间的差异不具有统计学意义(χ2=0.12,P>0.05)。
3.商品猪旋毛虫感染的强度使用人工消化法检疫18份阳性肉样时,共消化阳性肌肉样品122.8g,检出旋毛虫肌幼虫51条,阳性肉样的平均每克肌肉虫荷(1pg)为0.42(51/122.8),感染度较低。其中安阳的9份阳性肉样中平均每克肌肉虫荷为0.34(19/55.9),最低值为0.11,最高值为1.41；商丘的9份阳性肉样中均每克肌肉虫荷平均水平为0.48(32/66.9),最小值为0.10,最高值为1.58；二者的每克肌肉虫荷的最高值均超过了旋毛虫对公共卫生构成威胁的最低感染水平(1pg≥1)。对来自不同地方的阳性肉样的每克肌肉虫荷进行统计学分析,发现分别采集自安阳和商丘的阳性肉样虫荷之间的差异不具有统计学意义(Z=一0.05,P>0.05)。
4.多重PCR结果经过多重PCR扩增,我国河南安阳猪源旋毛虫分离株扩增出了一条大小为173bp的条带,该条带为旋毛虫(T1,ISS3)在多重PCR鉴定时的特有带型。河南安阳猪源旋毛虫分离株的电泳结果和河南南阳猪源地理株及旋毛虫(T1,ISS3)的结果一致,而不同于乡土旋毛虫(T2,127bp)、布氏旋毛虫(T3,127bp和253bp)、伪旋毛虫(T4,310bp)和纳氏旋毛虫(T7,155bp和404bp)。
结论
1.青海部分地区的屠宰猪未检出旋毛虫感染；河南省部分农村地区屠宰生猪的旋毛虫自然感染率较高,但感染强度较低。
2.河南省安阳地区猪源旋毛虫分离株属于旋毛形线虫(Trichinella spiralis, T1)。
Trichinellosis is an important food-borne parasitosis and parasitic zoonosis all over the world. When raw or undercooked meat or its products which contain infective Trichinella larvae were ingested, trichinellosis should occur. In recent years, outbreaks of human trichinellosis have taken place in many countries, and trichinellosis was defined as a re-emerging disease for its epidemiological trends. From 2004 to 2009, at least 15 outbreaks of trichinellosis were reported in China. Pork was the predominant source of outbreaks of human trichinellosis in China,12 out of 15 outbreaks were caused by eating raw or undercooked pork. Trichinellosis can cause expensive economic loss, not only harms to public health, but also to farming, especially the industrialized hoggery.
Eight species and 4 uncertain genotypes were recognised in the genus Trichinella till now. Because there were distinct differences among different species of Trichinella, for instance, morphology, geography distribution, the range of host, the resistance to freezing, the infecting ability to host and immunity response of host to different species, so the study on taxonomy of Trichinella is necessary. The aim of the study on Trichinella taxonomy is to offer valuable informations on the pathogen biology, the clinical treatment, the epidemiology, the prevention and control of trichinellosis in humans and animals. The range of Trichinella's hosts was very wide, and 8 species of Trichinella have been separated from wild animals. In China, only Trichinella spiralis (T1) and Trichinella nativa (T2) were reported till today, were there other species or genotypes of Trichinella exist in China still not sure.
In order to find the new Trichinella isolates in China, epidemiological investigation was carried on to separate novel isolates from Qinghai and Henan province in this study. The male Kunming mice were inoculated with new Trichinella isolates for species keeping. Further more, these different isolates of Trichinella were identified by multiplex PCR method. To provide the evidence for the relationship among pathogenesis, distribution and hosts, the epidemiology and molecular biology research were carried into execution in this study.
Materials and Methods
1. Epidemiological investigation The prevalence rates of swine trichinellosis in Qinghai and Henan Province were observed by epidemiological investigation. The sampling points were slected by local Animal Disease Control Center (ADC) and Center for Disease Control and Prevention (CDC). Samples were taken from diaphragm muscle and 50g each.
2. Samples inspection All the samples were numbered first, and quarantined by trichinelloscopy, then, all of them should be inspected again by artifical digestion method which recommended by International Commission on Trichinellosis (ICT).
3. Artificial inoculation of experimental animals Two month-old clean Kunming mice (18g～20g) were obtained from the Experimental Animal Center of Medical College of Zhengzhou University. After a whole day fast, male Kunming mice were orally inoculated with new Trichinella isolate from the same sampling point for species keeping.
4. Recovery of Trichinella muscle larvae The Kunming mice which infected with different isolates and international reference strains of Trichinella were killed 40 days later, and purified muscle larvae were collected by artifical digestion method.
5. Genomic DNA acquired and purity checked Genomic DNA was prepared according to phenyl-Chloroform method from muscle larvae. The purity of genomic DNA was checked by spectrophotometer (U-2001). OD value of A260 and A280 were detected by spectrophotometer first, and then the ratio of A260 to A280 was calculated for pure assessment. The purity of genomic DNA was fine when the ratio between 1.8-1.9.
6. Identification by multiplex PCR The genomic DNA which acquired from new isolate and different Trichinella international reference strains were identified by the method multiplex PCR. When the gene was amplied,5 pairs of primers that designed by Zarlenga (1999) were used. The products of multiplex PCR were electrophoresed in 2% agarose gel and visualised under Gene Genius Imaging System.
Results
1. The investigation on swine trichinellosis in Qinghai province In this investigation,192 pieces of meat was collected from diaphragm muscle in Huzu and Delingha of Qinghai province. Samples were quarantined by trichinelloscopy and digestion method, and none trichinella was found. All the samples were negative, none of the Trichinella infective swine was found in parts of Qinghai province.
2. The investigation on swine trichinellosis in Henan province In Henan province, sampling points were choosen from Anyang and Shangqiu city. Three groups of diaphragm muscle samples were collected from 278 swine. None Trichinella was found when quarantined by trichinelloscopy, but 18 positive samples were detected when the digestion method was used, the positive rate of all the samples was 6.47%(18/278). All the positive samples were collected from the swine that breeded in pig bed by local farmer, and the Trichinella infection rate of this kind of swine was 10.84%(18/166). The swine which from industrialized hoggery were free of Trichinella infection (0/112), and the differentiation was statistically significant (χ2=11.67, P<0.01) between local pig bed and industrialized hoggery of Trichinella infection rates. There was no difference of Trichinella infection rates in local pig bed between Anyang and Shangqiu (χ2=0.12,P>0.05).
3. Infective intensity of Trichinella in swine When 18 positive samples were detected by artifical digestion method, about 51 larvae were found from 122.8g of pork, and the larvae per gram (lpg) is 0.42. For the 9 positive samples that from Anyang county, the lpg has a range from 0.11 to 1.41, and the average level of lpg is 0.34 (19/55.9). Among the 9 Trichinella infective samples which from Sui county, the minimum of lpg is 0.10, the maximum is 1.58, with 0.48 (32/66.9) as its average. Both maxima of lpg are higher than the lowest level, which threaten to the public health (1pg≥1). The difference of lpg between Anyang county and Sui county without statistically significant (Z=-0.05, P>0.05).
4. Multiplex PCR Anyang isolate of Trichinella was identified by multiplex PCR method, and the result was observed by using electrophoresis. The Nanyang isolate and 5 Trichinella international reference strains were also analyzed as control at the same time. The special band (173bp) that belongs to Trichinella spiralis (T1, ISS3) was also showed by Nanyang and Anyang isolates, which was different from Trichinella nativa (T2,127bp), Trichinella britovi (T3,127bp and 253bp), Trichinella pesudospiralis (T4,310bp) and Trichinella nelsoni (T7,155bp and 404bp).
Conclusions
1. The swine infected with Trichinella are not found in part areas of Qinghai province. Though prevalence of swine trichinellosis is relatively high in rural part of Henan province, the intensity of infection is low.
2. The Trichinella isolates recovered from pork in Anyang were identified as Trichinella spiralis (T1).
引文
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