Clinical and pathologic features of experimental Schistosoma japonicum infection in pigs

Nine piglets, 2-5 months old and weighing 12-15 kg were infected percutaneously with 5000 to 6000 cercariae of Schistosoma japonicum. Three similar piglets which were not infected served as controls. Infected animals were necropsied at 5, 10, 30, 40, 59, 90, 120, 150 and 180 days post-infection (PI) and the controls at 5, 60 and 180 days PI. The prepatent period varied from 27 to 33 days after infection. Clinical signs observed, coincident with egg production, were loss of appetite, lethargy, pallor of mucous membranes, diarrhea, progressive emaciation and dehydration. One piglet was moribund when killed for necropsy at 40 days and another piglet died 59 days PI. Pathological changes induced by S. japonicum included erythematous papules on the site of inoculation, petechial hemorrhages in the lungs, catarrhal to hemorrhagic enteritis, bluish-gray discoloration of the liver, and egg granulomas in the liver, lungs, spleen, intestines, pancreas and mesenteric lymph nodes. Endophlebitis with intimal hyperplasia was occasionally observed in veins harboring adult schistosomes.     

Comparison of the vaccine efficacy of gamma-irradiated Schistosoma japonicum cercariae with the defined antigen Sj62(IrV-5) in pigs

Development of a vaccine against Schistosoma japonicum which can protect both man and the domestic animal zoonotic reservoirs of infection would be an invaluable tool in attempts to control this infection in those areas in which conventional control methods have failed to break transmission. The pig is a natural host of S. japonicum and because of its anatomical and immunological similarities to humans, it is a potentially valuable host for studies on S. japonicum in particular and schistosomes in general. Radiation-attenuated cercariae are highly effective in inducing immunity in experimental schistosomosis and there are promising reports of partial protection against schistosomes with recombinant-derived individual antigens. In the present study we have set out to establish a protocol for inducing protection with gamma-irradiated cercariae in pigs and to assess the protective capacity of recombinant and naked DNA formulations of Sj62, a 62kDa region of S. japonicum myosin. The corresponding S. mansoni version or Sj62, recombinant IrV-5, has previously been implicated in irradiated vaccine immunity in S. mansoni infections and has been shown to induce high levels of immunity in a variety of hosts.Groups of pigs were immunised three times at 2-week intervals with 2000 cercariae irradiated at 20krad, with Sj62 as a recombinant (rSj62) incorporated in Freund's adjuvant, a micellar preparation, or as a naked DNA construct. Vaccination with irradiated cercariae did not induce significant anti-Sj62 antibody but following intramuscular challenge with 2000 cercariae, the vaccinated pigs showed >95% resistance as assessed by reduced faecal egg output, worm tissue egg burdens and also reduced septal fibrosis. Immunisation with each of the Sj62 formulations induced significant anti-Sj62 antibody responses, the highest titre (>12,800) being with the Freund's preparation but none of the Sj62-immunised groups showed significant resistance to challenge. The data suggest that Sj62 shows little promise as a vaccine candidate for schistosomosis.     

Cytokine mRNA profiles in pigs exposed prenatally and postnatally to Schistosoma japonicum

The pig is a natural host for Schistosoma japonicum and a useful animal model of human infection. The aim of the present study was to assess the differences between the cytokine profiles in prenatally or postnatally S. japonicum exposed pigs. Seven prenatally exposed pigs, 7 postnatally exposed pigs and 4 uninfected control pigs were compared 27 weeks post exposure. Variables included worm burdens, tissue egg counts, liver pathology and mRNA levels of IL-2, IL-4, IL-10, IL-12, TNF-alpha, TGF-beta1 and IFN-gamma in the liver and the caecum, assessed by RT-PCR. Infection intensity and level of septal fibrosis were significantly higher in the postnatal group compared to the prenatal group (P < 0.05). A significant increase of IL-4 (P < 0.01), IL-10 (P < 0.01), IL-12 (P < 0.01) and TNF-alpha (P < 0.05) mRNA level was also observed in the caecum of prenatally infected animals compared to the control group (P < 0.01). The prenatal group showed higher levels of TGF-beta1 in the liver compared with the postnatally infected group (P< 0.05) and the control group (P< 0.01). This suppressive immune response correlated with previously reported low hepatic pathogenesis in prenatally exposed pigs.     

日本血吸虫蛋白质组学研究进展

随着日本血吸虫(Schistosoma japonicum)基因组学的发展,应用双向电泳、质谱分析及生物信息学分析开展的蛋白质组学研究正成为日本血吸虫研究的前沿领域。生物体中的基因是相对稳定,但是蛋白质的活动随着生命活动的进行表现出动态的变化,并具有对外界环境发生反应的能力。对日本血吸虫进行蛋白质组学方面的研究,能直接地反映其生理功能的过程。这些研究为研制开发抗日本血吸虫病疫苗、新型治疗药物提供了新的途径。论文从日本血吸虫童虫部分差异表达蛋白、成虫蛋白质的性别差异性、药物相关蛋白组学、免疫蛋白组学等方面,对日本血吸虫蛋白质组学研究做了概述。     

日本血吸虫非叉型尾蚴感染动物试验

试验采集云南巍山地区自然条件下的钉螺,压碎钉螺、检获尾蚴,发现为非叉型尾蚴。将非叉型尾蚴400~850条/只经皮肤感染家兔,同时,用云南洱源的叉型尾蚴作对照试验。感染后50～55 d解剖兔子,试验组检获到日本血吸虫,荷虫量为10～15条/只。试验结果表明,云南巍山非叉型尾蚴能感染动物,该试验结果为丰富日本血吸虫的生活史提供了极有价值的补充。     

Autoradiographic quantification of the efficacy of niridazole in mice infected with 75Se-labelled cercariae of Schistosoma mansoni.

The effects of the anti-schistosomal drug, niridazole, on the migration of Schistosoma mansoni larvae, biosynthetically radioisotope-labelled with 75[Se]-selenomethionine, was evaluated by autoradiography of compressed tissues of mice treated daily from Days 6 to 10 post-infection with 200 mg kg-1 niridazole. The results were compared with the migration of schistosomula in untreated controls. The distribution of schistosomula was altered in niridazole-treated mice, where there was a delayed migration from the lungs relative to the controls and significantly fewer schistosomula in total appeared to reach the liver. The total percentage of schistosomula detected as autoradiographic foci was significantly lower in treated mice than in the untreated controls. Niridazole-treated mice were free of any foci 10 days after the last treatment and no adult worms were recovered on perfusion of the hepatic portal system relative to control mice from which 5.8% of the infective cercariae were recovered as adult worms at Day 42 post-infection.     

日本血吸虫铁蛋白基因在油菜中的表达

根据GenBank日本血吸虫铁蛋白基因(SjFer)的序列设计1对特异性引物,利用PCR从cDNA文库中扩增SjFer基因,与T载体连接并测序。常规方法构建植物表达载体SjFer/pBI121,采用根癌农杆菌介导法转化油菜子叶,组织培养卡那霉素抗性筛选,获得转化植株。转化植株通过PCR和Northern-blot杂交进行鉴定。结果表明SjFer可在油菜中正确表达。该研究为进一步研究转基因油菜工程疫苗奠定了基础。     

阿魏倍半萜类对日本血吸虫作用研究

观察小鼠体内不同发育阶段日本血吸虫童虫与成虫对阿魏倍半萜类的敏感性。用定量日本血吸虫尾蚴感染小鼠,感染2 h,1、3、7、12、14、16、25、35、42 d后分别一次性灌服阿魏倍半萜类500 mg/kg。给药后4周剖检小鼠,采用门静脉灌注法收集虫体,计算减虫率。结果显示,阿魏倍半萜类对1、3、7、12、14、16、25、35、42 d龄童虫或成虫的减虫率分别为16.9%、18.0%、71.3%、50.2%、36.9%、31.3%、45.3%、58.0%、26.4%,其中以7-35 d龄虫对该药最敏感。该结果提示,阿魏倍半萜类对小鼠体内不同发育阶段日本血吸虫均有杀灭作用,对7-35 d龄虫体杀灭效果更为明显。     

HMGB1与日本血吸虫病肝炎症和纤维化的相关性分析

旨在探究高迁移率族蛋白B1（high mobility group box 1,HMGB1）与日本血吸虫病肝炎症和纤维化的相关性,本研究将BALB/c小鼠随机分为空白对照组（n＝10）,感染后4周组、6周组和HMGB1抑制剂组（n＝5）,进行血常规及生化检测,观察肝病理学及Masson染色情况、荧光定量PCR检测HMGB1、IL-1β、TNF-α、IL-6、IFN-γ、α-SMA、Col1α1、TGF-β1、Smad2和Smad3的转录水平。结果显示：感染后4周,小鼠谷丙转氨酶（alanine aminotransferase,ALT）、肝组织HMGB1、α-SMA、TGF-β1、IL-6和 TNF-α的转录水平均显著或极显著升高（P<0.05或P < 0.01）;感染后6周,小鼠WBC、Neu、Mon、Eos、谷草转氨酶（aspartate aminotransferase,AST）和ALT极显著升高（P<0.01）,肝组织HMGB1、IL-1β、TNF-α、IL-6、IFN-γ、α-SMA、Col1α1、TGF-β1、Smad2和Smad3的转录水平均显著或极显著升高（P<0.05或P<0.01）;感染后6周小鼠肝虫卵聚积处伴有炎性细胞浸润和胶原纤维增生;HMGB1抑制剂可显著降低小鼠血清ALT值和肝组织HMGB1、α-SMA、TGF-β1、IL-6和TNF-α的转录水平（P<0.05）。综上表明,HMGB1与日本血吸虫病肝炎症和纤维化具有一定相关性。     

不同剂量新疆阿魏对血吸虫活性的影响

为了探讨新疆阿魏杀灭血吸虫的功效,分别用新疆阿魏蒸馏萃取物以0.1、0.2、0.3、0.4 g/kg浓度药液灌服治疗感染血吸虫病家兔,每隔3 d灌服1次,4次后停止给药,4周后剖检,门静脉灌注法收集虫体、计算减虫率,并以组织切片检查该药对血吸虫感染兔肝脏的治疗作用。结果显示,减虫率分别为12.30%、17.20%、46.40%、40.80%;经组织切片观察,家兔肝脏肉芽肿明显减小。表明新疆阿魏具有很强的杀灭血吸虫活性的作用,最佳杀虫剂量为0.3 g/kg。     

3个日本血吸虫新基因的克隆和分析

为获得新的可表达保护性抗原分子的编码基因 ,以重复感染兔血清为探针 ,筛选日本血吸虫成虫 c DNA文库 ,对获得的阳性克隆进行 PCR鉴定并测序后 ,通过互联网将测序结果进行同源性分析 ,并对新基因的编码阅读框及其编码蛋白质的结构及功能进行预测。 3个日本血吸虫新基因 IR1、IR2、IR3在 Gen Bank的登录号分别为 AY1 70 31 3、AY1 73930、AY2 5 1 4 81。 BL AST分析结果表明 ,它们与 Gen Bank中其他基因无显著的同源性。其中 IR3为全基因 ,含897个碱基 ,编码 2 99个氨基酸 ,推测等电点 ( p I)为 6 .0 9,相对分子质量约为 331 90     

Parenteral immunization of pigs against infection with Treponema hyodysenteriae.

Six intravenous injections of formalin-inactivated Treponema hyodysenteriae were given to 8 specific-pathogen-free pigs at 6-day intervals. The 8 vaccinated and 8 control pigs were challenged intragastrically with pure cultures of T hyodysenteriae 7 and 8 days after the last intravenous injection. Clinical signs of swine dysentery were observed in all 8 control pigs, but was observed in only 1 of the immunized pigs. Three control pigs died. These findings suggest that parenteral immunization with T hyodysenteriae provided a marked degree of protection against subsequent intragastric challenge exposure with the homologous isolate of T hyodysenteriae.     

山羊日本血吸虫抗体消长规律

山羊７只各接种日本血吸虫尾蚴（３００±２０）条，２只作空白对照。粪孵阳性后，７只感染羊被随机分为２组，其中一组（４只）以吡喹酮按每日每公斤体重用１００ｍｇ，共用２ｄ进行治疗，另一组（３只）作对照。用环卵沉淀试验（ＣＯＰＴ）、间接血凝试验（ＩＨＡ）和酶联免疫吸附试验（ＥＬＩＳＡ）３种方法检测其抗体消长。结果，ＣＯＰＴ和ＩＨＡ于感染后第２０天检出特异性抗体，ＥＬＩＳＡ于第４０天检出特异性ＩｇＧ；经吡喹酮治疗后，上述３种方法分别于３，５，７个月转阴。     

Application of recombinant Sjc26GST for serodiagnosis of Schistosoma japonicum infection in water buffalo (Bos buffelus)

Schistosomiasis japonica is currently the most serious parasitic disease in mainland China and it is estimated that several million people are infected. Furthermore, it is also responsible for the deaths of many domestic animals. In order to establish an effective diagnostic method, the gene encoding Sjc26GST was cloned and expressed in Escherichia coli as a fusion protein with His-tag. The purified reSjc26GST was used as an antigen for an enzyme-linked immunosorbent assay (ELISA) and for immunoblotting detection of Schistosoma japonicum antibodies in water buffaloes. Our results showed that mean OD values of specific serum IgG antibodies from egg-positive buffaloes were 3.37-fold higher than what was found in egg-negative buffaloes from non-endemic areas. The data also showed the OD value of the endemic egg-negative group reached as high as 1.69 times as that found in non-endemic areas. The positivity rate of egg-positive buffaloes was 100%, but was 30.3% in the endemic egg-negative group. Infected bovine antisera also recognized reSjc26GST, a 27 kDa protein as determined by Western blot. These results suggest that the recombinant GST expressed in E. coli should be an effective diagnostic reagent for detection of antibody against S. japonicum in buffaloes. Due to straightforward production, excellent sensitivity and high specificity, the reSjc26GST described in this study can be considered as a candidate protein for immunological diagnosis of bovine schistosomiasis. Developing reSjc26GST, with its potential diagnostic values, will be useful for diagnosis and surveillance of schistosomiasis in controlling the spread of this parasitic disease in domestic animals.     

湖北省四湖水系牛日本血吸虫感染与再感染及其危险因素调查

为研究湖滩型日本血吸虫病中重度流行区化疗后牛群血吸虫感染、再感染及其危险因素,采用在观察试区,以34例感染牛经吡喹酮治疗后虫卵转为阴性的30头牛为"再感染"研究对象;98例粪检阴性牛群作为"感染"研究对象.追踪观察两组牛群在一个感染季节后的再感染情况,并调查其接触疫水情况以及当地螺情和疫水变化情况等凶素;再对其进行单因素分析和多因素非条件logistic回归分析.结果显示,基线感染率为20.26%,经过一个感染季节后,标准化感染率为10.01%,标准化再感染率高达20.62%,再感染率已恢复到化疗前牛群感染水平,其中黄牛再感染率(26.66%)高于水牛感染率(13.33%).接触疫水处水体尾蚴密度、年龄、6～9月疫水接触指数、牧地距阳性螺点距离是再感染发生的主要危险因素.接触疫水处水体尾蚴密度、牛采食和6～9月疫水接触指数是血吸虫病感染的主要危险因素.本研究表明,在日本血吸虫病重度流行区存在较快感染和再感染趋势,初步揭示了感染和再感染的主要危险因素.     

阿魏减轻小鼠血吸虫性肝纤维化试验

血吸虫病的病理基础是虫卵肉芽肿。无虫卵或虽有虫卵而不发生或少发生肉芽肿，则不发生血吸虫病或病情较轻。国内外学者发现，在肉芽肿形成中自由基O^-2起着重要作用，抗氧化剂能缩小肉芽肿体积。阿魏（Resina ferulae）是一种天然的抗氧化中药，具有抗肿瘤、降血脂等多种药理作用，但未见其抗血吸虫性肝纤维化的研究报道。笔者等用阿魏治疗试验小鼠血吸虫病，观察其对血吸虫性肝纤维化形成的影响。     

Enteric adenovirus infection in pigs.

Intranuclear adenovirus inclusion bodies were identified in intestinal epithelium by histological, electron microscopical or cytological examination of 43 conventionally raised pigs submitted for diagnostic evaluation from 38 farms between August 1981 and January 1983. The affected pigs were usually less than three weeks old, but ranged from five days to 24 weeks of age. The inclusion bodies occurred in the epithelium of ordinary intestinal villi, or more commonly, in the epithelium covering the blunt villi located over Peyer's patches in the ileum. Fifty-three percent of these adenovirus infected pigs had diarrhea. In most of these pigs with diarrhea (19 of 23) however, agents previously established as causes of porcine enteric disease were also identified. Of the pigs with enterocyte inclusion bodies but without diarrhea, the adenovirus infection appeared incidental to the major disease processes in the pigs. These findings suggest that this adenovirus is an asymptomatic and probably incidental infection in conventional farm-raised pigs.