Rossano M G, Schott H C, Murphy A J, Kaneene J B, Sellon D C, Hines M T, Hochstatter T, Bell J A, Mansfield L S
The Population Medicine Center, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA.
Vet Parasitol. 2005 Jan 4;127(1):3-8. doi: 10.1016/j.vetpar.2004.08.023.
Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in Americans. Most cases are attributed to infection of the central nervous system with Sarcocystis neurona. Parasitemia has not been demonstrated in immunocompetent horses, but has been documented in one immunocompromised foal. The objective of this study was to isolate viable S. neurona from the blood of immunocompetent horses. Horses used in this study received orally administered S. neurona sporocysts (strain SN 37-R) daily for 112 days at the following doses: 100/day for 28 days, followed by 500/day for 28 days, followed by 1000/day for 56 days. On day 98 of the study, six yearling colts were selected for attempted culture of S. neurona from blood, two testing positive, two testing suspect and two testing negative for antibodies against S. neurona on day 84 of the study. Two 10 ml tubes with EDTA were filled from each horse by jugular venipuncture and the plasma fraction rich in mononuclear cells was pipetted onto confluent equine dermal cell cultures. The cultures were monitored weekly for parasite growth for 12 weeks. Merozoites grown from cultures were harvested and tested using S. neurona-specific PCR with RFLP to confirm species identity. PCR products were sequenced and compared to known strains of S. neurona. After 38 days of in vitro incubation, one cell culture from a horse testing positive for antibodies against S. neurona was positive for parasite growth while the five remaining cultures remained negative for parasite growth for all 12 weeks. The Sarcocystis isolate recovered from cell culture was confirmed to be S. neurona by PCR with RFLP. Gene sequence analysis revealed that the isolate was identical to the challenge strain SN-37R and differed from two known strains UCD1 and MIH1. To our knowledge this is the first report of parasitemia with S. neurona in an immunocompetent horse.
马属动物原生动物脑脊髓炎(EPM)是美国马匹中一种严重的神经系统疾病。大多数病例归因于中枢神经系统被神经肉孢子虫感染。在免疫功能正常的马匹中尚未证实有寄生虫血症,但在一匹免疫功能低下的幼驹中已有记录。本研究的目的是从免疫功能正常的马匹血液中分离出活的神经肉孢子虫。本研究中使用的马匹每天口服神经肉孢子虫孢子囊(菌株SN 37 - R),持续112天,剂量如下:第1至28天每天100个,接下来28天每天500个,随后56天每天1000个。在研究的第98天,挑选了6匹一岁龄的雄驹尝试从血液中培养神经肉孢子虫,在研究的第84天,其中两匹抗神经肉孢子虫抗体检测呈阳性,两匹检测可疑,两匹检测呈阴性。通过颈静脉穿刺从每匹马采集两管10 ml含乙二胺四乙酸(EDTA)的血液,将富含单核细胞的血浆部分移液到汇合的马皮肤细胞培养物上。每周监测培养物中寄生虫生长情况,持续12周。从培养物中生长的裂殖子被收获,并使用具有限制性片段长度多态性(RFLP)的神经肉孢子虫特异性聚合酶链反应(PCR)进行检测,以确认物种身份。对PCR产物进行测序,并与已知的神经肉孢子虫菌株进行比较。在体外培养38天后,一匹抗神经肉孢子虫抗体检测呈阳性的马的一个细胞培养物中寄生虫生长呈阳性,而其余5个培养物在全部12周内寄生虫生长均为阴性。通过具有RFLP的PCR确认从细胞培养物中分离出的肉孢子虫为神经肉孢子虫。基因序列分析表明,分离株与攻击菌株SN - 37R相同,与两个已知菌株UCD1和MIH1不同。据我们所知,这是免疫功能正常的马匹中出现神经肉孢子虫寄生虫血症的首次报告。