兔脑炎原虫肾细胞培养物不同成分抗原性比较

兔脑炎原虫病血清学诊断方法的研究在国外报道较多,在我国则刚刚起步,目前用于各种方法的抗原均来自培养提纯的脑炎原虫,而脑炎原虫有无分泌抗原,培养物中哪些成分抗原性最强,至今在国内尚未见报道,为此我们用BA—ELISA方法对培养物的不同成分进行了比较试验,以促进本病诊断方法的发展。     

屠宰兔脑炎原虫病的快速检验

脑炎原虫病是一种人畜共患的寄生虫病,其病原为小孢子虫目(Microsporidia)、微粒子虫科(Nosematae)的兔脑炎原虫(En-cephalitozoon Cuniculi)。病原侵害的主要靶器官为脑组织和肾脏。据报道脑炎原虫的感染范围较广,可使鼠、兔、犬、猫、猴、     

实验兔脑炎原虫病的临床症状及流行病学特点

兔脑炎原虫又名脑孢子虫 (Encephalitozooncuniculi,Nicolau和 Schoen,1 92 3) ,是一种细胞内寄生的原虫。关于脑炎原虫的分类地位目前存在较大争议 ,暂列在原生动物界、顶器门、孢子虫纲、微孢子虫目。兔脑炎原虫具有广泛的宿主范围 ,包括无脊椎动物 (昆虫 )、啮齿类动物、兔形动物、草食动物、肉食动物、禽类和灵长类动物等 ,人类亦可以感染。近十几年来有大量有关兔脑炎原虫引起爱滋病患者致死性感染的临床报道 ,故本病引起医学界的广泛重视。国外有关人脑炎原虫病的病理变化、致病机理、诊断技术方面等做了大量的研究。在动物中 ,脑炎…     

兔脑炎原虫病肾脏病变的病理形态学观察

对97只患脑炎原虫病獭兔的肾脏病变做了系统的病理形态学研究.眼观,可将肾脏病变分为三型:皱缩型、凹陷型和斑点型;镜检,依结缔组织增生的数量、部位和肾组织结构的不同改变而分为贯通型、局灶型和肉芽肿型.     

兔脑炎原虫的分离与鉴定

3只临床表现斜颈、麻痹和打滚的病兔,经病理组织学检查诊断为脑炎原虫病.取其脑组织制成乳剂,接种于兔肾细胞、Vero细胞、猫肾细胞和兔脉络丛细胞.用前3种细胞均分离出原虫.该虫体表现为多种形态,呈杆状、圆形或卵圆形;Gram染色呈阳性,Goodpasture氏染色呈红色.用20%H_2O_2处理虫体,在相差显微镜下见到虫体极丝的伸展.通过虫体的形态学、染色特征和极丝的突出实验,鉴定该虫体为脑炎原虫.兔肾细胞、Vero细胞和猫肾细胞的感染率分别为40%、40%和5%,而在兔脉络丛细胞原代培养物中未检查到虫体.     

Immunohistochemical identification of Encephalitozoon cuniculi in phacoclastic uveitis in four rabbits

PURPOSE: Encephalitozoon cuniculi is a microsporidium with a wide range of mammalian hosts. In rabbits it can be responsible for cataract and lens-induced uveitis (LIU). The aim of this study was to provide specific immunohistochemical demonstration and localization of E. cuniculi within the eye, in rabbits with LIU. MATERIAL AND METHODS: Four rabbits were presented with a white mass in the eye and iris discoloration. Complete ophthalmic examinations were performed and a presumptive diagnosis of LIU was made in all cases. Initial therapy with a topical steroid, atropine and systemic enrofloxacin was instituted while serologic (IFA or ICA tests) and cytologic lab results were pending. The final outcome in all cases was enucleation. Routine histology and immunohistochemistry (ABC method) with an antiserum anti-Encephalitozoon cuniculi were performed. RESULTS: Indirect immunofluorescence performed on one rabbit serum expressed a titer of 1 : 32; carbon immunoassay on the serum of the other three rabbits expressed a titer of 1 : 5120 in one, and a titer of 1 : 2560 in the other two cases. Histologically, an intraocular, locally extensive pyogranulomatous infiltration that partially filled the posterior chamber, encasing a wide anterior lens capsule break, was detected in all cases. Immunohistochemically, spores reacting with anti-Encephalitozoon cuniculi antiserum were present in all specimens, occasionally within macrophages and lens epithelial cells. CONCLUSION: Detection of E. cuniculi in rabbits with phacoclastic uveitis has been investigated in the past with different methods. Based on our results, we suggest that immunohistochemistry should be regarded as a useful tool both for specific demonstration of E. cuniculi and for its localization within tissues.     

兔脑炎原虫的超微形态与发育

用电镜连续７代动态地观察了培养细胞中兔脑炎原虫的超微结构与发育状态。脑炎原虫含有细胞核，但无线粒体、内质网和高尔基复合体等细胞器，其表面有几根极丝。脑炎原虫在培养细胞中除二分裂增殖外，还有裂体增殖和配体增殖。由于脑炎原虫有３种增殖方式，故在光学显微镜下呈多形态结构。     

Serological survey for antibodies to Encephalitozoon cuniculi in pet rabbits in Italy

Pet rabbits (n = 125) from Southern Italy were submitted to a serological screening for Encephalitozoon cuniculi, using an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA). Seventy-eight examined rabbits showed clinical signs suggestive of encephalitozoonosis (head tilt, ataxia, paralysis, cataracts, uveitis, polyuria and polydipsia), whereas 47 were healthy rabbits. Antibodies anti-E. cuniculi were found in 84/125 (67.2%) sera analysed. The results of the chi-squared test showed that sex and health status had no significant effect (P > 0.05) on E. cuniculi seropositivity; however, rabbits older than 4 months had a seropositivity for E. cuniculi significantly (P < 0.05) higher than that of rabbits aged up to 4 months. The results of the present survey reinforce the assumption that rabbit may be indicated as the main reservoir of E. cuniculi; therefore, routine screening examinations in pet rabbits are strongly advised considering the zoonotic potential of this parasite.     

阿苯达唑对自发性兔脑炎原虫病的治疗

为了有效地治疗兔脑炎原虫病,本研究根据脑炎原虫的生物学特点和对组织损伤的特征选用阿苯达唑进行了治疗试验。将28只3～4月龄隐性感染的獭兔,随机分为对照组和治疗组进行试验。治疗组按30mg／kg剂量（首次给药量为50mg／kg）,每隔12h给药1次,连续用药10d,停药1周为1个治疗期,共治疗3个疗程。对照组按正常饲养。每1疗程之后,均采血和尿液进行ELISA检测和尿沉渣虫体检查。结果,治疗组的病兔用阿苯达唑治疗3个疗程后,ELISA检测的D值均低于标准值（〈0．044）,即抗脑炎原虫血清抗体呈阴性反应,从尿沉渣中也不能检出虫体。而对照组病免的D值则大大高于正常值．从尿沉渣中均易检出虫,并有1只对照兔出现典型的神经症状,剖检后从脑组织中检出脑炎原虫性肉芽肿。经方差分析,2组间差异非常显著（P〈0．01）。试验证明,阿苯达唑对兔脑炎原虫有良好治疗作用,用药的时机以脑炎原虫还未进入脑组织时最合适,用药的剂量一定要保持有效的杀虫浓度。     

Microsporidia (Encephalitozoon cuniculi) in wild rabbits in Australia

Objective To determine the prevalence of infection with Encephalitozoon cuniculi i n wild rabbit populations in Western Australia, and to isolate the organism from seropositive rabbits.
Design Serological screening of wild and clinically affected domestic rabbit populations.
Sample population Eighty-one wild rabbits from southwestern Western Australia and 29 laboratory rabbits.
Procedure Indirect immunofluorescence antibody technique and in-vitro amplification of parasite isolates in fibroblast cultures.
Results Of the 81 wild rabbits and 29 laboratory rabbits, 20 and 22 respectively, had antibodies to E cuniculi . E cuniculi from the urine of one seropositive laboratory rabbit and from brain and kidney tissues of eight and five seropositive laboratory and wild rabbits respectively were isolated in fibroblast cultures.
Conclusion E cuniculi infection has been shown for the first time to be prevalent in wild rabbits in Australia. Techniques have been developed for the isolation and culture of the causative agent. Comparative studies can now be undertaken to determine risk factors for clinical disease in domestic rabbits and the relationship among E cuniculi isolates from wild and domestic rabbits.     

IL-2在自然感染兔脑炎原虫獭兔体内免疫作用的研究

为观察 IL-2在自然感染兔脑炎原虫獭兔体内的免疫作用 ,用双抗体夹心 EL ISA试剂盒对 8只自然感染兔脑炎原虫的獭兔和 4只健康长耳白对照兔的血清及培养 3 d和 4d的脾和淋巴结的细胞培养上清中的 IL-2进行了检测。检测结果为病兔与对照兔 IL -2平均含量差异不显著 (P >0 .0 5) ,但病兔 IL -2平均含量高于对照兔。提示 IL -2对病兔产生了一定的免疫作用 ,但可能不扮演主要角色     

脑炎原虫病兔免疫器官的病理形态学观察

研究脑炎原虫病兔外周免疫器官的形态学变化,采集20只病兔和12只对照兔的淋巴结、脾脏和圆小囊,通过临床检查,病理剖检,病理切片的HE染色、吉姆萨染色和免疫组织化学染色进行了比较观察.结果表明病兔淋巴结内的淋巴小结数量增多,生发中心明显,髓索增粗,副皮质区增宽.脾脏的脾小结增多,变大,具有明显的生发中心,边缘区明显增宽,动脉周围淋巴鞘明显变大,聚集大量淋巴细胞.圆小囊的淋巴小结增大,数量增多,生发中心明显,淋巴小结周围弥散大量淋巴细胞.通过α-ANAE和sABC染色,在淋巴结、脾脏和圆小囊中均可检出大量T细胞、巨噬细胞和IgG阳性细胞.据此认为患脑炎原虫病时,病兔的体液免疫和细胞免疫均增强,但以细胞免疫反应更强.     

Detection of Encephalitozoon cuniculi in the feline cataractous lens

Purpose Identification of Encephalitozoon cuniculi (E. cuniculi) as a possible causative agent for cataracts and uveitis in cats. Methods Within a 12‐month study period, cats that were presented with focal anterior cortical or mature cataract and secondary uveitis underwent a complete ophthalmic examination, complete blood count, serum biochemistry, serologic tests for E. cuniculi and tests for feline immunodeficiency virus (FIV), feline infectious peritonitis (FIP), feline leukemia virus (FeLV) and Toxoplasma gondii (T. gondii). PCR for DNA detection of E. cuniculi and T. gondii as well as cytologic examination of aqueous humor after paracentesis and phacoemulsified lens material were also performed. In addition histopathologic examination of the resected anterior lens capsule and attached lens epithelial cells was performed. Serologic testing for antibodies against E. cuniculi was also performed in 100 ophthalmologically healthy cats. Results Eleven (19 eyes) European shorthair cats with a median age of 3.5 years were included. Nine of 11 cats had bilateral cataracts, with 12/19 eyes having focal anterior cortical cataracts and 7/19 eyes having mature cataracts. In 14/19 eyes anterior uveitis was present. All cats had a positive antibody titer (1:80–1:10 000) for E. cuniculi. Encephalitozoon cuniculi DNA was detected by PCR and sequencing in 18/19 lenses and in 10/19 aqueous samples. Five tentative positive results were detected by cytologic examination. Spores were detected in 15/19 samples of lens material with histopathologic staining. Only 2/100 ophthalmologically healthy cats showed a positive antibody titer for E. cuniculi. Conclusion Encephalitozoon cuniculi is a cause of focal anterior cortical cataract and anterior uveitis in cats.