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Abstract：
BACKGROUND: Toxoplasma gondii and Neospora caninum are closely related protozoan parasites that are considered important causes of abortion in livestock, causing huge economic losses. Hunan Province ranks 12th in the production of beef and mutton in China. However, limited data are available on the seroprevalence, risk factors and molecular characterization of T. gondii and N. caninum in beef cattle and goats in Hunan province, China.
METHODS: Sera of 985 beef cattle and 1147 goats were examined for the presence of specific antibodies against T. gondii using indirect hemagglutination test (IHAT) and anti-N. caninum IgG using competitive-inhibition enzyme-linked immunoassay assay (cELISA). Statistical analysis of possible risk factors was performed using PASW Statistics. Muscle samples of 160 beef cattle and 160 goats were examined for the presence of T. gondii DNA (B1 gene) and N. caninum DNA (Nc-5 gene) by nested PCR. The B1 gene-positive samples were genotyped at 10 genetic markers using the multilocus nested PCR-RFLP (Mn-PCR-RFLP).
RESULTS: Specific IgG against T. gondii were detected in 8.3% (82/985) and 13.3% (153/1147) and against N. caninum in 2.1% (21/985) and 2.0% (23/1147) of the beef cattle and goats, respectively. Based on statistical analysis, the presence of cats, semi-intensive management mode and gender were identified as significant risk factors for T. gondii infection in beef cattle. Age was a significant risk factor for T. gondii infection in goats (P < 0.05), and age > 3 years was a significant risk factor for N. caninum infection in beef cattle (P < 0.05). PCR positivity for T. gondii was observed in three beef samples (1.9%; 3/160) and seven chevon samples (4.4%; 7/160). Genotyping of PCR positive samples identified one to be ToxoDB#10. The N. caninum DNA was observed in one beef sample (0.6%; 1/160) but was negative in all chevon samples.
CONCLUSIONS: To our knowledge, this is the first large-scale serological and molecular investigation of T. gondii and N. caninum and assessment of related risk factors in beef cattle and goats in Hunan Province, China. The findings provide baseline data for executing prevention and control of these two important parasites in beef cattle and goats in China.
METHODS: Sera of 985 beef cattle and 1147 goats were examined for the presence of specific antibodies against T. gondii using indirect hemagglutination test (IHAT) and anti-N. caninum IgG using competitive-inhibition enzyme-linked immunoassay assay (cELISA). Statistical analysis of possible risk factors was performed using PASW Statistics. Muscle samples of 160 beef cattle and 160 goats were examined for the presence of T. gondii DNA (B1 gene) and N. caninum DNA (Nc-5 gene) by nested PCR. The B1 gene-positive samples were genotyped at 10 genetic markers using the multilocus nested PCR-RFLP (Mn-PCR-RFLP).
RESULTS: Specific IgG against T. gondii were detected in 8.3% (82/985) and 13.3% (153/1147) and against N. caninum in 2.1% (21/985) and 2.0% (23/1147) of the beef cattle and goats, respectively. Based on statistical analysis, the presence of cats, semi-intensive management mode and gender were identified as significant risk factors for T. gondii infection in beef cattle. Age was a significant risk factor for T. gondii infection in goats (P < 0.05), and age > 3 years was a significant risk factor for N. caninum infection in beef cattle (P < 0.05). PCR positivity for T. gondii was observed in three beef samples (1.9%; 3/160) and seven chevon samples (4.4%; 7/160). Genotyping of PCR positive samples identified one to be ToxoDB#10. The N. caninum DNA was observed in one beef sample (0.6%; 1/160) but was negative in all chevon samples.
CONCLUSIONS: To our knowledge, this is the first large-scale serological and molecular investigation of T. gondii and N. caninum and assessment of related risk factors in beef cattle and goats in Hunan Province, China. The findings provide baseline data for executing prevention and control of these two important parasites in beef cattle and goats in China.
摘要：
背景：弓形虫和犬新孢子虫是密切相关的原生动物寄生虫，被认为是牲畜流产的重要原因，造成巨大的经济损失。湖南省在中国牛羊肉生产中排名第12位。然而,关于血清阳性率的数据有限，湖南省肉牛和山羊弓形虫和犬奈瑟菌的危险因素及分子特征,中国。
方法：使用间接血凝试验（IHAT）和抗N。检查了985头肉牛和1147头山羊的血清中是否存在针对弓形虫的特异性抗体。使用竞争抑制酶联免疫测定(cELISA)的犬IgG。使用PASWStatistics对可能的危险因素进行统计分析。通过巢式PCR检查了160头肉牛和160只山羊的肌肉样品中是否存在弓形虫DNA（B1基因）和犬硝虫DNA（Nc-5基因）。使用多位点巢式PCR-RFLP（Mn-PCR-RFLP）对B1基因阳性样品的10个遗传标记进行基因分型。
结果：在8.3%（82/985）和13.3%（153/1147）的肉牛和山羊中检测到了针对弓形虫的特异性IgG，在2.1%（21/985）和2.0%（23/1147）分别。根据统计分析,猫的存在，半集约化管理模式和性别是肉牛弓形虫感染的重要危险因素。年龄是山羊弓形虫感染的显著危险因素(P<0.05),年龄>3岁是肉牛犬根虫感染的重要危险因素（P<0.05）。在三个牛肉样品（1.9％；3/160）和七个chevon样品（4.4％；7/160）中观察到弓形虫的PCR阳性。PCR阳性样品的基因分型鉴定为ToxoDB#10。在一个牛肉样品(0.6%;1/160)中观察到犬奈瑟菌DNA,但在所有chevon样品中是阴性的。
结论：据我们所知，这是湖南省首次对肉牛和山羊进行大规模的弓形虫和犬奈瑟菌的血清学和分子学调查，并对相关危险因素进行评估，中国。研究结果为在中国肉牛和山羊中执行这两种重要寄生虫的预防和控制提供了基线数据。
方法：使用间接血凝试验（IHAT）和抗N。检查了985头肉牛和1147头山羊的血清中是否存在针对弓形虫的特异性抗体。使用竞争抑制酶联免疫测定(cELISA)的犬IgG。使用PASWStatistics对可能的危险因素进行统计分析。通过巢式PCR检查了160头肉牛和160只山羊的肌肉样品中是否存在弓形虫DNA（B1基因）和犬硝虫DNA（Nc-5基因）。使用多位点巢式PCR-RFLP（Mn-PCR-RFLP）对B1基因阳性样品的10个遗传标记进行基因分型。
结果：在8.3%（82/985）和13.3%（153/1147）的肉牛和山羊中检测到了针对弓形虫的特异性IgG，在2.1%（21/985）和2.0%（23/1147）分别。根据统计分析,猫的存在，半集约化管理模式和性别是肉牛弓形虫感染的重要危险因素。年龄是山羊弓形虫感染的显著危险因素(P<0.05),年龄>3岁是肉牛犬根虫感染的重要危险因素（P<0.05）。在三个牛肉样品（1.9％；3/160）和七个chevon样品（4.4％；7/160）中观察到弓形虫的PCR阳性。PCR阳性样品的基因分型鉴定为ToxoDB#10。在一个牛肉样品(0.6%;1/160)中观察到犬奈瑟菌DNA,但在所有chevon样品中是阴性的。
结论：据我们所知，这是湖南省首次对肉牛和山羊进行大规模的弓形虫和犬奈瑟菌的血清学和分子学调查，并对相关危险因素进行评估，中国。研究结果为在中国肉牛和山羊中执行这两种重要寄生虫的预防和控制提供了基线数据。
