Verma S K, von Dohlen A Rosypal, Mowery J D, Scott D, Rosenthal B M, Dubey J P, Lindsay D S
United States Department of Agriculture, Agricultural Research Service, Beltsville Agricultural Research Center, Animal Parasitic Diseases Laboratory, Building 1001, Beltsville, Maryland 20705-2350.
J Parasitol. 2017 Oct;103(5):555-564. doi: 10.1645/17-10. Epub 2017 Jun 23.
Here, we report a new species of Sarcocystis with red-tailed hawk (RTH, Buteo jamaicensis) as the natural definitive host and IFN-γ gene knockout (KO) mice as an experimental intermediate host in which sarcocysts form in muscle. Two RTHs submitted to the Carolina Raptor Center, Huntersville, North Carolina, were euthanized because they could not be rehabilitated and released. Fully sporulated 12.5 × 9.9-μm sized sporocysts were found in intestinal scrapings of both hawks. Sporocysts were orally fed to laboratory-reared outbred Swiss Webster mice (SW, Mus musculus) and also to KO mice. The sporocysts were infective for KO mice but not for SW mice. All SW mice remained asymptomatic, and neither schizonts nor sarcocysts were found in any SW mice euthanized on days 54, 77, 103 (n = 2) or 137 post-inoculation (PI). The KO mice developed neurological signs and were necropsied between 52 to 68 days PI. Schizonts/merozoites were found in all KO mice euthanized on days 52, 55 (n = 3), 59, 61 (n = 2), 66, and 68 PI and they were confined to the brain. The predominant lesion was meningoencephalitis characterized by perivascular cuffs, granulomas, and necrosis of the neural tissue. The schizonts/merozoites were located in neural tissue and were apparently extravascular. Brain homogenates from infected KO mice were infective to KO mice by subcutaneous inoculation and when seeded on to CV-1 cells. Microscopic sarcocysts were found in skeletal muscles of 5 of 8 KO mice euthanized between 55-61 days PI. Only a few sarcocysts were detected. Sarcocysts were microscopic, up to 3.5 mm long. When viewed with light microscopy, the sarcocyst wall appeared thin (<1 μm thick) and smooth. By transmission electron microscopy, the sarcocyst wall classified as "type 1j" (new designation). Molecular characterization using 18S rRNA, 28S rRNA, ITS-1, and cox1 genes revealed a close relationship with Sarcocystis microti and Sarcocystis glareoli; both species infect birds as definitive hosts. The parasite in the present study was biologically and molecularly different from species so far described in RTHs and we therefore propose a new species name, Sarcocystis jamaicensis n. sp.
在此,我们报告一种新的肉孢子虫物种,其天然终末宿主为红尾鹰(RTH,赤肩鵟,Buteo jamaicensis),实验中间宿主为IFN-γ基因敲除(KO)小鼠,在其肌肉中可形成肉孢子囊。提交至北卡罗来纳州亨特斯维尔市卡罗来纳猛禽中心的两只红尾鹰因无法康复并放归自然而实施安乐死。在两只鹰的肠道刮片中均发现了大小为12.5×9.9μm、已完全完全完全成熟的孢子囊。将孢子囊经口投喂给实验室饲养的远交系瑞士韦伯斯特小鼠(SW,小家鼠,Mus musculus)以及KO小鼠。孢子囊对KO小鼠具有感染性,但对SW小鼠无感染性。所有SW小鼠均未出现症状,在接种后第54、77、103天(n = 2)或137天处死的任何SW小鼠中均未发现裂殖体或肉孢子囊。KO小鼠出现神经症状,并在接种后52至68天进行尸检。在接种后第52、55天(n = 3)、59、61天(n = 2)、66和68天处死的所有KO小鼠中均发现裂殖体/裂殖子,且均局限于脑部。主要病变为脑膜脑炎,其特征为血管周围套袖、肉芽肿以及神经组织坏死。裂殖体/裂殖子位于神经组织中,显然位于血管外。感染的KO小鼠的脑匀浆经皮下接种以及接种到CV-1细胞上后,对KO小鼠具有感染性。在接种后55 - 61天处死的8只KO小鼠中的5只的骨骼肌中发现了显微镜下可见的肉孢子囊。仅检测到少数肉孢子囊。肉孢子囊在显微镜下可见,最长可达3.5毫米。在光学显微镜下观察,肉孢子囊壁似乎很薄(<1μm厚)且光滑。通过透射电子显微镜观察,肉孢子囊壁分类为“1j型”(新命名)。使用18S rRNA、28S rRNA、ITS-1和cox1基因进行分子鉴定,结果显示与微小肉孢子虫(Sarcocystis microti)和格氏肉孢子虫(Sarcocystis glareoli)关系密切;这两个物种均以鸟类作为终末宿主。本研究中的寄生虫在生物学和分子水平上与迄今在红尾鹰中描述过的物种不同,因此我们提出一个新的物种名称,即牙买加肉孢子虫(Sarcocystis jamaicensis n. sp.)