家畜的住肉孢子虫与住肉孢子虫病

家畜住肉孢子虫病是一种人畜共患的寄生虫病。住肉孢子虫最初是Miescher(1843)在欧洲的小鼠肌肉中发现的,其后在其它动物的肌肉中也相继发现。Kuhn(1865)在猪肉中发现了该原虫,并以Miescher 的名字命名为Synchytium mies-cheriahum。Lankester(1882)把本种作为模式种,新设Sarcocystis 属。住肉孢子虫住分类上属原生动物亚界、原生动物门、孢子虫纲、真球虫     

棱形住肉孢子虫和巨型住肉孢子虫抗原的免疫特性研究

动用ＣＩＥ和ＥＬＩＳＡ对两种住肉孢子虫全包裹抗原及包裹各部分抗原的免疫特性进行了分析研究。结果表明：两种住肉孢子虫与网种或异种住肉孢子虫阳性血清具有强烈的交叉反应,各囊各部分抗原以缓殖子抗原免疫反应最强。     

梭形住肉孢子虫和巨型住肉孢子虫抗原的免疫特性研究

运用 ＣＩＥ和 ＥＬＩＳＡ对两种住肉孢子虫全包裹抗原及包裹各部分抗原的免疫特性进行了分析研究;结果表明;两种住肉孢子虫与同种或异种住肉孢子虫阳性血清具有强烈的交叉反应,包囊各部分抗原以缓殖子抗原免疫反应最强．     

人畜共患住肉孢子虫病

人畜共患住肉孢子虫是一种寄生在人和动物的肌肉或肠道中的原生动物寄生虫，需要在两个不同种的宿主循环，才能完成其生活史。无性发育阶段通常发生在中间宿主完成，有性发育阶段在终末宿主完成。林氏住肉孢子虫人为其中间宿主，人住肉孢子虫人是终末宿主。本文根据文献和自己的研究成果综述了人畜共患住肉孢子虫发现史、形态生活史、发病机制、症状体征与治疗、人和动物流行病学及防治措施。     

住肉孢子虫病防治试验报告

住肉孢子虫系二宿主寄生物，其所致病性与防治工作同发育史密切相关。近些年来，国外许多学者依据虫体发育史特点，应用免疫、药物平方法在小范围内或通过实验感染进行了防治试验研究，获得了不同程度的效果；有的观察和介绍了温度对中间宿主胴体内住肉孢子虫存活期的影响等。国内仅有报道人作为终末宿主的肠壁型肉孢子虫病的防治试验。家畜住肉孢子虫病的药物防治试验尚未见有报道。作者仅就天然感染和人工感染的绵羊住肉孢子虫病病例作动物模型，进行了药特防治试验。     

黄牛住肉孢子虫病

住肉孢子虫Sarcocystis隶属原生动物门顶器亚门孢子虫纲真球虫目球虫亚目肉孢子虫科(Levine,1973),广泛分布世界各地,对黄牛有很高的感染率,过去曾一直被认为对牛无致病性,但近十多年的研究     

家畜住肉孢子虫感染情况调查

对北京,河北,内蒙,云南,湖南等地家畜住肉孢子虫的感染情况进行了调查,黄牛,水牛,绵羊,山羊和猪的平均感染率分别为４３．５％、９１％、５６．６％、５９．６％和１３．８％。调查还发现,同一畜种不同地方的感染率有着明显差异。在不同饲养方式下对住肉孢子虫的感染率也明显不同。     

家畜住肉孢子虫研究进展

住肉孢子虫是一类细胞内的寄生性原虫,广泛发现于爬行类、鸟类和包括人、猴、鲸、各种家畜在内的哺乳动物。根据Ｌｅｖｉｎｅ（１９８６）统计,文献记载的虫种有１２０多种,其中已知生活史的有５６种,寄生在家畜有２０多种,而近１０年来仍不断有新种报道。７０年代生...     

Sarcocystis sp. encephalomyelitis in a cat

Abstract: A 5‐month‐old male neutered domestic shorthair cat was evaluated for spinal pain, ataxia, and anisocoria. Neuroanatomic localization indicated diffuse or multifocal central nervous system disease. On cerebrospinal fluid analysis, neutrophilic pleocytosis and intracellular protozoal merozoites were observed. The merozoites were oval, 2–4 μm in width and 4–6 μm in length, and had linear arrays of nuclear material concentrated at one pole. Serum was positive for Sarcocystis sp. antibodies and negative for Toxoplasma gondii antibodies. The organism was determined to be either Sarcocystis neurona or Sarcocystis dasypi based on sequence analysis of the internal transcribed spacer 1 ribosomal RNA genomic region. Clinical disease resolved following treatment with 3 different protocols for protozoal infection. This case is the first to demonstrate the antemortem diagnosis and survival of a domestic cat with Sarcocystis sp.‐associated encephalomyelitis. Clinicians and cytopathologists should include Sarcocystis sp. as a differential for feline inflammatory central nervous system disease characterized by neutrophilic pleocytosis.     

牛环形泰勒虫enolase基因的原核表达及生物信息学分析

【目的】获得牛环形泰勒虫(Theileria annulata)新疆株enolase基因,并分析其生物学特性及反应原性。【方法】对牛环形泰勒虫enolase基因进行扩增和克隆,构建原核表达载体pGEX-4T-1-enolase,诱导表达enolase重组蛋白并进行蛋白纯化,通过Western blotting验证enolase重组蛋白反应原性。利用生物信息学方法对enolase基因编码蛋白的理化性质、亲疏水性、跨膜区、信号肽、磷酸化、亚细胞定位及蛋白互作网络进行预测分析。【结果】PCR扩增出大小为1 248 bp的牛环形泰勒虫enolase基因片段,enolase重组蛋白大小约70 ku; Western blotting结果表明,该重组蛋白与牛环形泰勒虫阳性血清发生反应。enolase基因编码416个氨基酸,理论等电点为5.91,有38个磷酸化位点;二级结构主要由α-螺旋(43.03%)和无规则卷曲(33.17%)组成;具有17个B细胞抗原表位,亚细胞定位主要位于细胞质中;enolase蛋白与磷酸甘油酸变位酶(PGAM)、二磷酸核苷激酶(NDK)、磷酸甘油酸激酶(PGK)、热休克蛋白...     

Morphology of Sarcocystis gigantea in experimentally-infected sheep

The development of the parasite and lesions was studied in 32 sheep killed 10 days to 47 months after inoculation with Sarcocystis gigantea sporocysts from cats. At 21-42 days post-inoculation (d.p.i.), there was a mild encephalitis, but organisms were not seen in the brain. Immature sarcocysts were detected from 40-84 d.p.i. The cyst wall was not measurable by light microscopy at 40 d.p.i., but was 1.5-2 microns thick at 84 d.p.i. At 119 d.p.i. both immature cysts containing only metrocytes, and mature cysts containing both metrocytes and merozoites, were present. These mature cysts did not have a secondary cyst wall. A mature cyst, 350 microns in length, was found in a sheep killed at 8 1/2 months p.i. At 10 m.p.i. cysts were up to 0.5 mm long and a secondary cyst wall was present. At 47 m.p.i. cysts were 2-5 X 4.5-7.5 mm, and were found only in the muscles of tongue, oesophagus, pharynx and flank.     

The in vitro excystation of Sarcocystis gigantea sporocysts

Studies on the in vitro excystation of Sarcocystis gigantea sporocysts revealed that pretreatment before exposure to trypsin and bile was an essential prerequisite. However, in contrast to Sarcocystis tenella and Sarcocystis capracanis, incubation in cysteine hydrochloride under CO2 was largely unsuccessful for excysting Sarcocystis gigantea: of the pretreatments tested, only exposure to sodium hypochlorite proved effective. Excystation from sodium hypochlorite-pretreated S. gigantea sporocysts took place in trypsin and bile between temperatures of 30 and 43 degrees C and occurred rapidly at 39 degrees C. While the presence of bile or bile salts was essential for this process, that of trypsin was not, although more sporocysts excysted in its presence than in its absence. Excystation occurred in the presence of all bile types tested but not when Tween 80 was substituted for bile. The highest levels of excystation were recorded when cattle or sheep bile or sodium taurocholate were used and the lowest when chicken or pig bile were employed. Neither the concentration of sheep bile above 2.5%, nor hydrogen ion concentration (pH range 5.0-10.0) appeared to have any marked effect on the level of excystation obtained.     

驴肉孢子虫包囊的超微结构及其实验感染

对昆明地区的26头驴进行了肌肉检查，在24头驴肌肉中发现了肉孢子虫包囊，其然自感染率为92．3％，光镜下包囊呈梭形，具有锥状和棍棒状的突起，透射电镜下突起内微管在顶端松散排列，向下延伸集结成束，斜伸入基质层，有的和母细胞的膜相连，用含有包囊的驴肉实验感染犬和猫各2只，感染后在犬粪便中发现孢子囊和卵囊，潜隐期为11－13 d，剖检在2只犬的小肠固有层中查到孢子囊和卵囊，猫粪便中一直未检到孢子囊和卵囊，剖检亦未发现，表明驴肉孢了虫的终末宿主要是犬，而是猫，研究认为驴全内的肉孢子虫仅有1种，鉴定为柏氏肉孢子虫（Sarcocystis bertrami).     

Effects of Enolase of Streptococcus equi ssp. zooepidemicus on Phagocytic Functions of Alveolar Macrophages in Mice

To investigate the effect of enolase (Eno) of Streptococcus equi ssp. zooepidemicus (SEZ) on phagocytosis of mouse alveolar macrophages (RAW264.7). Recombinant enolase (rEno) was obtained by constructing prokaryotic expression plasmid, and the cytotoxicity of rEno protein on RAW264.7 cell proliferation was determined by trypan-blue living cell count method. After the rEno protein was incubated with RAW264.7 cells, SEZ was applied to the cells and the quantity of bacteria being phagocytosed was detected to determine the phagocytic activity of RAW264.7 cells. Further, candidate proteins that might interact with SEZ Eno in RAW264.7 cells were screened by live cell stable isotope labeling (SILAC) and protein spectrum analysis (LC-MS/MS). It was found that protein treatment (rEno,10 μg·mL^-1) had significant cytotoxic effects on RAW264.7 cells. Treatment of RAW264.7 cells with 10.0 μg·mL^-1 rEno protein for 2 and 4 hours could significantly inhibit the phagocytosis of RAW264.7 cells (P<0.01, P<0.05). In RAW264.7 cells, dynactin subunit protein 2 (Dctn), integrin alpha-M and about 17 proteins that might interact with Eno were preliminarily identified as rEno interaction proteins. The rEno recombinant expression protein was obtained in this study, and it could reduce the phagocytosis of RAW264.7 cells to SEZ. Preliminary screening of interacting proteins also laid a foundation for further revealing the mechanism of Eno in the anti-phagocytosis of SEZ.     

马链球菌兽疫亚种烯醇化酶对小鼠肺泡巨噬细胞吞噬功能的影响

旨在研究马链球菌兽疫亚种（Streptococcus equi ssp.zooepidemicus,SEZ）烯醇化酶（enolase,Eno）对小鼠肺泡巨噬细胞（RAW264.7）吞噬能力的影响。通过构建原核表达质粒获得重组烯醇化酶（rEno）,采用台盼蓝活细胞计数法,判定在不同处理浓度和时间下rEno蛋白对RAW264.7细胞的细胞毒性。将rEno蛋白与RAW264.7细胞共孵育后,用SEZ作用于细胞并检测细胞吞菌数量,判断RAW264.7细胞对SEZ的吞噬活性。进一步通过活细胞稳定同位素标记技术（SILAC）和蛋白质谱分析技术（LC-MS/MS）,筛选到RAW264.7细胞中可能与SEZ Eno存在相互作用的候选蛋白。结果发现,10 μg·mL^-1 rEno蛋白处理对RAW264.7细胞有明显的细胞毒性,且10 μg·mL^-1 rEno蛋白处理RAW264.7细胞2和4 h可显著抑制其对SEZ的吞噬作用（P<0.01、P<0.05）。初步筛选到RAW264.7细胞中动力蛋白激活蛋白亚单位蛋白（dynactin subunit protein 2,Dctn）、整合素α-M蛋白（integrin alpha-M）等17种可能与Eno发生互作的蛋白。本研究获得了rEno重组表达蛋白,发现rEno可减少RAW264.7细胞对SEZ的吞噬,互作蛋白的初步筛选也为进一步揭示Eno在SEZ抗吞噬中的作用机制奠定了基础。     

河南省部分地区奶牛肉孢子虫感染的血清学调查

为了解河南省奶牛肉孢子虫病流行情况,2018年3月至2019年9月用间接酶联免疫吸附试验(ELISA)对采自河南省13个地区58个奶牛场的1 819份血样进行了肉孢子虫抗体检测分析.结果肉孢子虫抗体阳性牛场31个,场阳性率53.4％,阳性样品424份,样品平均阳性率为23.3％;不同生长阶段牛群阳性率差异极显著(P＜0...     

The in vivo excystation of Sarcocystis gigantea and S. tenella sporocysts

In vivo studies on the excystation of Sarcocystis gigantea and S. tenella sporocysts indicated that this process was, as in vitro, a diphasic one involving both pretreatment and treatment phases. The studies also tended to support in vitro observations that the requirements for the excystation of these two species are quite different. The results suggested that for neither species was the pretreatment stimulus likely to be provided by conditions in the rumen alone. However, exposure to abomasal conditions only induced moderate levels of excystation in both when they were subsequently treated with trypsin and bile. For S. gigantea, 0.25 to 4 h abomasal exposure was most effective; for S. tenella, 24 hours. The stimuli necessary to complete the excystation process could, apparently, be provided by 1 h placement in the duodenum for S. gigantea but not for S. tenella.