Verma S K, Lindsay D S, Mowery J D, Rosenthal B M, Dubey J P
U.S. 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):547-554. doi: 10.1645/17-2. Epub 2017 Jun 15.
Here, we report a new species, Sarcocystis pantherophisi n. sp., with the Eastern rat snake (Pantherophis alleghaniensis) as natural definitive host and the interferon gamma gene knockout (KO) mouse as the experimental intermediate host. Sporocysts (n = 15) from intestinal contents of the snake were 10.8 × 8.9 μm. Sporocysts were orally infective to KO mice but not to laboratory-raised albino outbred house mice (Mus musculus). The interferon gamma KO mice developed schizont-associated neurological signs, and schizonts were cultivated in vitro from the brain. Mature sarcocysts were found in skeletal muscles of KO mice examined 41 days postinoculation (PI). Sarcocysts were slender, up to 70 μm wide and up to 3.5 mm long. By light microscopy, sarcocysts appeared thin-walled (<1 μm) without projections. By transmission electron microscopy, the sarcocyst wall was a variant of "type 1" (type 1i, new designation). The parasitophorous vacuolar membrane (pvm) had approximately 100-nm-wide × 100-nm-long bleb-like evaginations interspersed with 100-nm-wide × 650-nm-long elongated protrusions at irregular distances, and invaginations into the ground substance layer (gs) for a very short distance (6 nm). The gs was smooth, up to 500 nm thick, without tubules, and contained a few vesicles. Longitudinally cut bradyzoites at 54 days PI were banana-shaped, 7.8 × 2.2 μm (n = 5). Molecular characterization using 18S rRNA, 28S rRNA, ITS-1, and cox1 genes indicated a close relationship with other Sarcocystis parasites that have snake-rodent life cycles. The parasite in the present study was molecularly and biologically similar to a previously reported isolate (designated earlier as Sarcocystis sp. ex Pantherophis alleghaniensis) from P. alleghaniensis, and it was structurally different from other Sarcocystis species so far described.
在此,我们报告一种新物种——豹纹蛇肉孢子虫(Sarcocystis pantherophisi n. sp.),以东美鼠蛇(Pantherophis alleghaniensis)作为天然终末宿主,以干扰素γ基因敲除(KO)小鼠作为实验中间宿主。从蛇的肠道内容物中获取的子孢子囊(n = 15)大小为10.8×8.9μm。子孢子囊经口感染KO小鼠,但不感染实验室饲养的白化远交家鼠(小家鼠Mus musculus)。干扰素γ基因敲除小鼠出现与裂殖体相关的神经症状,并且从脑中体外培养出了裂殖体。在接种后41天(PI)检查的KO小鼠骨骼肌中发现了成熟的肉孢子囊。肉孢子囊细长,宽达70μm,长达3.5mm。通过光学显微镜观察,肉孢子囊壁薄(<1μm)且无突起。通过透射电子显微镜观察,肉孢子囊壁是“1型”的变体(1i型,新命名)。寄生泡膜(pvm)有间隔不规则的约100nm宽×100nm长的泡状外翻,其间散布着100nm宽×650nm长的细长突起,并且向基质层(gs)内陷很短的距离(6nm)。基质层光滑,厚度达500nm,无小管,且含有一些小泡。接种后54天纵向切开的缓殖子呈香蕉形,大小为7.8×2.2μm(n = 5)。使用18S rRNA、28S rRNA、ITS-1和cox1基因进行的分子特征分析表明,该寄生虫与其他具有蛇 - 啮齿动物生命周期的肉孢子虫寄生虫关系密切。本研究中的寄生虫在分子和生物学方面与先前报道的来自东美鼠蛇的分离株(先前命名为Sarcocystis sp. ex Pantherophis alleghaniensis)相似,并且在结构上与迄今为止描述的其他肉孢子虫物种不同。