PCR技术在住肉孢子虫虫种鉴定上的应用

采用 RAPD 方法对水牛梭形住内孢子虫、待定种及黄牛枯氏住内孢子虫缓殖子基因组 DNA 进行了分析,待定种 PCR 产物的电泳带型与枯氏住肉孢子虫的差异较梭形住内孢子虫大,这与二者表型性状一致的研究结果不吻合.对3种包囊种特异性的 PCR 指纹进行了筛选,回收和纯化了3种包囊种特异性的 DNA 片段,待定种0.7kb 片段经α-~(32)PdATP 标记后用作探针,只与同源 DNA 杂交,不与其它包囊、宿主细胞和艾美耳球虫杂交,用 pGEM-T Easy Vector 对该片段克隆成功,并测出其全长序列,为进一步设计特异引物建立标准 PCR 诊断方法和用作核酸探针奠定了基础。     

Epidemiological investigations of trichostrongylid infections in young cattle in different parts of Norway

The occurrence of trichostrongylid nematodes in young cattle was studied in relation to climate and geographical location, by examination of herbage and faecal samples from 8 farms in 7 different regions in Norway, during 3 consecutive summers. Species of Ostertagia and Cooperia were dominant. Infective larvae, especially those of Ostertagia, were able to survive the winter in the pasture to a great extent in all the areas studied. The spring larval counts varied considerably from place to place, and from year to year, but were often at fairly high levels. This confirms previous findings that the larvae which overwinter on the pasture may be a major cause of losses. Overt clinical disease, however, was only seen at 1 farm during this investigation.The large number of larvae often seen in the autumn seems to be of less significance as a cause of disease, due to the late appearance, but is of major importance as a source of the larvae which survive the winter in the pasture and infect young cattle the following spring.There was no obvious correlation between pasture larval counts and the climate, except that there seemed to be higher counts in the warmer parts of the country, where the grazing season is also longer. The only place were both faecal egg counts and larval counts in the herbage were consistently low, was in the very far north.     

水牛梭形住肉孢子虫抗原的薄层等电聚焦行为分析

应用薄层聚丙烯酰胺等电聚焦电泳（ＩＦＥ）对水牛梭形住肉孢子虫（Ｓａｒｃｏｃｙｓｔｉｓｆｕｓｉｆｏｒｍｉｓ）抗原进行了分析。结果表明，经ＳｅｐｈａｄｅｘＧ－１００纯化的包囊抗原在电泳图谱上显示３条相距较近的带，等电点分别为６．３０，６．４５，６．６５；缓殖子纯化抗原仅出现１条带，等电点为６．４０     

中国水牛两种住肉孢子虫包囊超微结构的比较研究

本文对采自我国湖南水牛食道肌的大、小两种住肉孢子虫包囊的超微结构进行了研究。两种包囊的细胞(母细胞和缓殖子)未发现有明显的结构区别,而包囊壁的结构则揭示出明显的差异。大包囊的原囊壁向表面皱褶形成菜椰花样的突起,突起内含纤丝,突起表面有凹陷;而小包囊的原囊壁皱褶形成长指形的突起,突起内不含纤丝,倒伏在包囊表面,突起壁是平滑的,无凹陷。这一观察表细:我国水牛体内的大包囊属于梭形住肉孢子虫,小包囊可能是枯氏住肉孢子虫。     

Age variations in the prevalence of sarcocystosis in sheep and goats from northern and central Jordan

Awassi sheep and mountain goats slaughtered in northern and central Jordan were examined for Sarcocystis cysts by post-mortem examination, trichinoscopy of muscle samples taken from esophagi and diaphragms and serologically by indirect hemagglutination (IHA) during the period June 1986–May 1988. Macrosarcocysts were found in 11.3% (70/620) of sheep and 11.7% (46/393) of goats aged over 1 year old. Microsarcocysts were found in 50.1% (1185/2693) of the sheep diaphragm samples and 56.4% (711/1261) of the same organs of goats. The prevalences were lower in the esophageal muscles (26.4%, 348 of 1319 and 25.1%, 97 of 386) than the diaphragmal muscles (29.0%, 383 of 1319 and 34.2%, 132 of 386) in the younger age groups (less than 7 months) of sheep and goats, respectively. The prevalences in both the esophagus and the diaphragm increased with age. The seroprevalences of anti-Sarcocystis antibodies in sheep and goats were significantly higher (P < 0.05) than the prevalence of Sarcocystis cysts detected by trichinoscopy. The seroprevalence of sarcocystosis increased with age in sheep and goats.     

巨型住肉孢子虫烯醇酶基因克隆与表达

巨型住肉孢子虫(Sarcocystis gigantea)是羊体内一种常见寄生虫,多寄生于羊横纹肌内形成包囊,导致羊肉大量废弃,给畜牧业带来巨大经济损失。本研究尝试筛选能有效诊断住肉孢子虫感染的抗原。通过免疫印迹及质谱分析筛选到巨型住肉孢子虫候选诊断抗原烯醇酶,利用染色体步移技术扩增两侧翼未知序列并表达重组蛋白。应用生物信息学分析和免疫印迹对该蛋白诊断价值进行评价。结果筛选到的候选蛋白为巨型住肉孢子虫烯醇酶(SgENO),克隆得到1 181 bp的基因并获得重组蛋白rSgENO。对rSgENO应用进行初步评价,发现其与其他顶复亚门原虫的烯醇酶氨基酸相似性均较高(71%～92.1%),且能被弓形虫和新孢子虫阳性血清所识别,具有交叉反应性。本研究克隆并表达了巨型住肉孢子虫烯醇酶,后期评价发现该重组蛋白与新孢子虫和弓形虫有较强的交叉反应性,不能用于羊住肉孢子虫的血清学诊断,但有成为疫苗候选分子的潜力。     

水牛实验性感染枯氏住肉孢子虫病血液生化指标的变化

分别用1×10~7,5×10~8,2×10~6和8×10~5剂量的枯氏住肉孢子虫Sarcocystis cruzi孢子囊感染7头公水牛犊,感染后5～8周,血清谷草转氨酶(SGOT)活性和血液尿素氮(BUN)含量升高;血清谷丙转氨酶(SGPT)活性除5×10~6剂量组外均呈下降;血清乳酸脱氢酶(SLDH)活性升高,但8×10~5剂量组升高不明显;血清碱性磷酸酶(SAKP)活性以及血清钠、钾、镁和钙的含量试验组与对照组之间无明显差异;血清氯的含量随感染剂量而变化,即l×10~7和5×10~6剂最组感染后5周升高,而8×10~5剂量组在整个试验期间均卞降;血清磷变化轻微且与感染剂量有关。     

Equine Protozoal Myeloencephalitis: An Updated Consensus Statement with a Focus on Parasite Biology,Diagnosis, Treatment,and Prevention

Equine protozoal myeloencephalitis (EPM) remains an important neurologic disease of horses. There are no pathognomonic clinical signs for the disease. Affected horses can have focal or multifocal central nervous system (CNS) disease. EPM can be difficult to diagnose antemortem. It is caused by either of 2 parasites, Sarcocystis neurona and Neospora hughesi, with much less known about N. hughesi. Although risk factors such as transport stress and breed and age correlations have been identified, biologic factors such as genetic predispositions of individual animals, and parasite‐specific factors such as strain differences in virulence, remain largely undetermined. This consensus statement update presents current published knowledge of the parasite biology, host immune response, disease pathogenesis, epidemiology, and risk factors. Importantly, the statement provides recommendations for EPM diagnosis, treatment, and prevention.     

Detection of Zoonotic Protozoa Toxoplasma gondii and Sarcocystis suihominis in Wild Boars from Spain

Food safety regulations require the control of the presence of protozoa in meats destined for human consumption. Wild boar (Sus scrofa) meat may constitute a source of zoonoses. A 23.8% (688/2881) seroprevalence of anti‐Toxoplasma gondii antibodies and 72.2% (662/910) Sarcocystis sarcocysts prevalence were detected among wild boars hunted in Southwestern areas of Spain. Identity of Sarcocystis spp. was performed by RFLP‐PCR and sequencing, detecting S. miescheriana (7/8) and the zoonotic S. suihominis (1/8). Risk assessment studies of these coccidian in meats destined to human consumption are needed.