Comparative antigenic characterisation of Echinococcus granulosus and Taenia hydatigena cyst fluids by immunoelectrophoresis.

Hydatid cyst fluid from Echinococcus granulosus (HCF) and cyst fluid from Taenia hydatigena (TCF) cysts were compared in reciprocal immunoelectrophoresis (IEP) tests using homologous and heterologous antisera which were free of antibodies to host serum contaminants. The antigens for the E granulosus arc 5 were demonstrated in TCF. Antibody activity to these and other antigens common to HCF and TCF was removed from homologous antisera by absorptions with the heterologous antigenic preparation. Antigens not shared by the two metacestodes fluids were then demonstrated by IEP tests. These findings are discussed in terms of their significance to phylogenetic and immunodiagnostic studies of these parasites in their immediate hosts.     

Specific antibody responses to Taenia hydatigena, Taenia pisiformis and Echinococcus granulosus infection in dogs

Groups of dogs reared free of both nematodes and cestodes were infected with Taenia hydatigena, Taenia pisiformis or Echinococcus granulosus. After infections with the Taenia spp became patent, dogs were purged to remove the worms. They were later reinfected and the second infections again removed by purging after patency. A group of 3 uninfected worm free dogs was kept as age-matched controls. The dogs were bled at intervals of 5 days and their serums tested for antibodies using the enzyme-linked immunosorbent assay (ELISA) with excretory/secretory (ES) antigens collected during in vitro incubation of evaginated scoleces (scolex ES antigen) and oncosphere antigens. Antibodies to scolex ES antigen were detected by 3 weeks after infection with each cestode species whereas antibodies to oncosphere antigen were not detected until about one week after eggs were found in the faeces of the infected dogs. Antibody responses to both oncosphere and scolex ES antigens decreased rapidly following removal of the worms by purging. Uninfected control dogs were invariably negative to both oncospheral and scolex ES antigens. There were cross-reactions between the serums from dogs infected with T. pisiformis and T. hydatigena when tested with scolex ES antigens, but oncospheral antigens showed a high degree of species specificity. Scolex ES antigens from E. granulosus were compared with those prepared from T. hydatigena and T. pisiformis for their ability to discriminate between antibodies in serums collected from dogs 31 and 32 days after infection with 100,000 protoscoleces of E. granulosus or dogs infected with Taenia spp.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of fospirate on Echinococcus granulosus and Taenia hydatigena infections in dogs.

Trials with fospirate carried out with 170 dogs infected with either Echinococcus granulosus or Taenia hydatigena indicated that a dose rate of 40 mg/kg given on three occasions and 10 mg/kg given on one occasion eliminated each species, respectively. Increasing the dose rat substantially above 40 mg/kg did not reduce the number of treatments required to eliminate all hydatid organisms. Vomiting was observed in some dogs.     

The effect of mebendazole on Echinococcus granulosus and Taenia hydatigena infections in dogs.

The effect of micronised mebendazole on Echinococcus granulosus and Taenia hydratigena has been investigated in a trial involving 175 dogs. The hydatid organism was eliminated from the dogs at dose rates of either 160 mg/kg given on one occasion or 20 mg/kg given on two occasions. A single dose of 20 mg/kg eliminated T hydratigena. The drug was readily accepted when given in food and there were no untoward sequelae.     

The effect of nitroscanate on Echinococcus granulosus and Taenia hydatigena infections in dogs.

Trials with nitroscanate carried out on 186 dogs infected with either Echinococcus granulosus or Taenia hydatigena indicated that a single treatment at a dose rate of 1000 mg/kg or two treatments at 250 mg/kg eliminated the former, but a single treatment at 64 mg/kg was sufficient to eliminate the latter. At this dosing schedule, vomiting and diarrhoea occurred, as well as a transient 'tranquillising' effect on some dogs.     

The effect of oxfendazole on Echinococcus granulosus and Taenia hydatigena infections in dogs

Trials with oxfendazole carried out on 120 dogs infected with either Echinococcus granulosus or Taenia hydatigena indicated that a single treatment significantly reduced the proportion of dogs infected with tapeworms. Diarrhoea occurred occasionally when the dose rate exceeded 20 mg/kg.     

Echinococcus granulosus,Taenia hydatigena,T. ovis: Evaluation of cyst fluids as antigen for serodiagnosis of larval cestodes in sheep

In order to monitor the progress of New Zealand's hydatids eradication campaign, a specific, serological, diagnostic test is required to identify infected sheep. An indirect haemagglutination test, using pyruvic aldehyde-stabilized sheep erythrocytes as the antigen carrier, was developed for the serodiagnosis of larval cestode infections of sheep. Using cyst fluid from Echinococcus granulosus, Taenia hydatigena and T. ovis as the antigens in this test, it was shown that the larval cestode species, responsible for an infection in sheep harbouring a single specific infection, could be identified by the higher titre given with the homologous antigen, in comparison to that given with the heterologous antigens. Sera of sheep infected with two or more species were also tested by this method, and the only specific infections to be diagnosed by differential titres were those due to the presence of live E. granulosus cysts. These antigens cannot be used for the diagnosis of specific larval cestode infections in the field because of the cross-reactivity between cyst fluids. However, the test did show that infection with larval cestodes could be diagnosed on a non-specific basis.     

The effect of nitroscanate tablets of Echinococcus granulosus and Taenia hydatigena infections in dogs

Tablets of micronised nitroscanate (nominal particle size 2--3 microns) were given to a total of 190 dogs that had been experimentally infected with either Echinococcus granulosus or Taenia hydatigena. The efficiency of the drug in tablet form in freeing dogs from tapeworms, was investigated. The dose rate at which 50 per cent of normally fed dogs can be expected to be freed from E granulosus was found to be 89 mg/kg (95 per cent confidence limits 55 mg/kg to 140 mg/kg). The 90 per cent effective dose rate was not determined within the range 32 mg/kg to 250 mg/kg. The dose rate at which 90 per cent of normally fed dogs can be expected to be freed from T hydatigena was 37 mg/kg (95 per cent confidence limits 23 mg/kg to 60 mg/kg).     

The importance of the New Zealand campaign against Echinococcus granulosus and Taenia hydatigena on the prevalence of Taenia ovis

Extract

An important aspect of the campaign in New Zealand directed against Echinococcus granulosus and Taenia hydatigena is its possible effect on the prevalence of Taenia ovis. The burying of offal to eliminate E. granulosus and T. hydatigena is recommended as being preferable to cooking it and feeding it to dogs. The drastic reduction in the number of European rabbits (Oryctolagus cuniculus) in many districts has also virtually eliminated an important dog-food. Consequently, in addition to commercial products, many farmers are relying on more sheep carcass-meat than previously as a source of dog-food. Other things being equal, conditions would seem ideal for an increase in the prevalence of. T. ovis. This is particularly significant to New Zealand which depends so much on a meat export trade. Damage to this on account of T. ovis could in turn pre- judice a campaign directed against E. granulosus and T. hydatigena.     

Some observations on the differences in incidences between Echinococcus granulosus and Taenia hydatigena in the livers of sheep in New Zealand

Abstract

Extract

The object of this report is to record observations on the differences in incidence between Echinococcus granulosus and Taenia hydatigena in the livers of sheep of different age groups in New Zealand. The method of collection of data, examination, and classification of animals in these surveys have previously been described (Gemmell, 1961a Gemmell, M. A. 1961a. N.Z. vet. J., 9: 29–29. [Taylor &; Francis Online] , [Google Scholar], 1961b Gemmell, M. A. 1961b. N.Z. vet. J., 9: 37–37. [Taylor &; Francis Online] , [Google Scholar]).     

Evaluation of an antigenic fraction of Taenia hydatigena metacestode cyst fluid for immunodiagnosis of bovine cysticercosis

An antigenic fraction (ThFAS) isolated from Taenia hydatigena metacestode cyst fluid was used in an ELISA to detect antibodies to T saginata in experimentally and naturally infected cattle. In 10 calves given 1,000 to 100,000 T saginata eggs (20% to 60% viability), IgG and IgM antibodies were detected in all the calves by post-inoculation week 3. Immunoglobulin G antibody values remained increased until calves were slaughtered at post-inoculation weeks 13 to 26. Six naturally infected calves (determined by postmortem examination) were considered positive, using the ELISA. Shared antigens were demonstrated between ThFAS and T saginata and T crassiceps; there were no shared antigens between ThFAS and Haemonchus contortus or Fasciola hepatica. Specific lectin binding to ThFAS indicated the presence of glycoconjugates. Immunoblot analysis indicated that a low molecular weight polypeptide (10,000 Mr) bears the immunodiagnostic antigen.     

Serological diagnosis of Echinococcus granulosus infection in sheep using cyst fluid antigen processed by antibody affinity chromatography

Serum antibody responses in sheep naturally or experimentally infected with Echinococcus granulosus and/or other larval cestodes were examined using an enzyme-linked immunosorbent assay (ELISA) with various antigens prepared from sheep hydatid cyst fluid ( SHCF ). Serum donors included: sheep experimentally infected with E. granulosus and their age-matched non-infected controls; sheep experimentally infected with other helminth parasites; sheep naturally infected with E. granulosus both from Tasmania and the Australian mainland; sheep from Tasmania naturally infected with larval cestodes other than E. granulosus; and naturally reared sheep completely free from infection with larval cestodes. Attempts were made to eliminate serological reactions which were not specific for E. granulosus by using a series of antibody affinity chromatography steps to deplete crude SHCF antigen; these included adsorption with a monoclonal antibody, 3EgH 29-2, removal of host IgG using rabbit anti-sheep IgG antibody, and removal of antigens which bound non-specifically to normal sheep immunoglobulin. The final affinity-depleted antigen product was designated AD SHCF . Specific serological reactivity in infected sheep was very low. Affinity depletion of SHCF using 3EgH 29-2 did not appear to increase the specificity of serological diagnosis of E. granulosus infection when experimentally infected sheep were compared with their non-infected controls provided the latter were age-matched with experimental animals. The other affinity adsorption steps significantly reduced non-specific background binding to antigen by normal sheep serum. Despite this reduction in background in the ELISA, only low levels of antibody could be detected in naturally-infected sheep.(ABSTRACT TRUNCATED AT 250 WORDS)     

Use of Echinococcus granulosus worm antigens for immunodiagnosis of E. granulosus infection in dogs.

Echinococcus granulosus worm excretory/secretory antigens (WES) were used in ELISA for diagnosis of E. granulosus infection in dogs and compared with protoscolex somatic antigens (PSM). Sera from 224 dogs were tested. There was no correlation between ELISA absorbance values and E. granulosus worm burdens using either antigen. There was a significant linear relationship between absorbance values of sera tested in the ELISA using WES (W-ELISA) and the ELISA using PSM (P-ELISA). However, there was a small but significant difference between the absorbance values of the sera tested against the two antigens. Western blot analysis of WES using sera from E. granulosus-infected and uninfected dogs revealed antigenic components of relative molecular mass (Mr) larger than 94,000, Mr 94,000-68,000 and Mr 43,000-39,000 in worms, and these were specific for E. granulosus and not identified in PSM; these antigenic differences may be responsible for differences in reactivity in ELISA. The sensitivities of W-ELISA and P-ELISA were 80.8% and 75.6%, respectively. The specificities of W-ELISA and P-ELISA were 93.7% and 97.9%, respectively. The reduced specificity in W-ELISA was mainly attributable to increased background reactivity of sera from Taenia hydatigena-infected dogs. Despite the reduction in specificity, both ELISAs are valuable epidemiological tools to determine the prevalence of antibody to E. granulosus in dog populations and to monitor the success of hydatid control campaigns.     

Use of two polysaccharide antigens in ELISA for the detection of antibodies to Echinococcus granulosus in sheep sera

Polysaccharide antigens were obtained from either the secretions produced during in vitro cultivation of Echinococcus granulosus protoscoleces or from mouse hydatid cyst membranes by phenol extraction. When either of these antigens was used in an enzyme-linked immunosorbent assay antibody activities were detected in sera from sheep infected 27 or more weeks earlier with at least 100 E granulosus eggs. These antibody responses were significantly higher (P less than 0.05) than those of sheep infected with Taenia hydatigena or T ovis and tested with the E granulosus antigens. Very high cross-reacting antibody responses in sera from sheep recently infected with T hydatigena were only detected with the protoscoleces secretions antigen. Neither antigen was sufficiently sensitive or specific for serodiagnostic use. However, when sera were first tested with one antigen and then with the other, and only sera that were positive in both tests were regarded as positive, the overall sensitivity and specificity of this two antigen method increased to about 80 per cent.     

泡状带绦虫线粒体部分基因克隆及序列比对分析

采集甘肃兰州市区宠物犬体内两株泡状带绦虫,运用分子生物学方法对其线粒体部分基因进行克隆,并使用基因分析软件Larsergene V7.1进行序列分析,结果发现这两株虫体同源性达到99.6%,与国内的泡状带绦虫广东虫株和甘肃虫株同源性分别达到98.7%和98.4%,鉴定为泡状带绦虫;与其他带科绦虫同源性均高于85%,低于88%。其中,与国内牛带绦虫同源性最高,为88%;与国内巨颈带绦虫同源性最低,只有85.1%。推断不同带科绦虫的cox1基因种间差异明显,在带科绦虫的分类上意义较大。     

兔豆状囊尾蚴囊液蛋白的纯化与抗原性鉴定

为了找出抗原性强、免疫原性好的兔豆状囊尾蚴抗原蛋白,本研究对兔豆状囊尾蚴的头节、体节和囊膜等结构性抗原与囊液中代谢性抗原进行了比较,发现囊液代谢性抗原的免疫效果优于结构性抗原。通过SDS-PAGE和微孔超滤分离法对囊液蛋白进行纯化,再用纯化的蛋白免疫小鼠,制备高免血清,并对免疫鼠的免疫器官进行病理学和免疫组织化学检查。结果证明用从囊液中纯化的63 000蛋白免疫效果最好。用纯化的63 000蛋白免疫小鼠可获得高效价抗血清,小鼠免疫器官活化,巨噬细胞、滤泡树突细胞、淋巴细胞和浆细胞等各种免疫细胞增多,用免疫组织化学染色,免疫器官中的CD4、CD8T细胞和CD20B细胞呈强阳性反应。总之,从囊液中分离纯化的63 000蛋白抗原性强、免疫原性,可用于对豆状囊尾蚴病的免疫学诊断。     

Mitosis and differentiation in T-cells under cytotoxic action of Echinococcus granulosus hydatid fluid

In the T-cell line, D10, thymidine uptake was used to measure the proportion of cells in S-phase, and the MTT assay to measure the number of viable cells. The effect of Echinococcus granulosus hydatid fluid (HF) on the lymphocytes was assayed in 3-day cultures of the T-cell line, D10, in increasing concentrations of HF. Apparent cytotoxic effects of HF were recorded as a log-linear decline in S-phase activity, which was reduced by the presence of IL-1, IL-2, or a combination of the two. In the presence of IL-2, however, mitogenic treatment with concanavalin A increased the cytotoxic effect in 3-day cultures, while in day-2 cultures, HF itself showed mitogenic effect. HF-induced decline in S-phase activity was not matched by a parallel decline in viable cells, suggesting that the apparent cytotoxicity of HF could result from cell-cycle arrest. Depending on its origin, HF enhanced membrane expression of CD25 and CD38 on human peripheral blood lymphoblasts, and diminished that of CD28. Taken together, these changes suggest that HF can induce T-cell mitosis and reduce co-stimulation with subsequent T-cell anergy or apoptosis.