Vaccination of bovines against Echinococcus granulosus (cystic echinococcosis)

Hydatid disease is an important human zoonosis. Humans become infected from carnivores that are infected with the Echinococcus granulosus tapeworm. Carnivores become infected after consuming hydatid cysts from grazing animals, which are generally sheep, goats and cattle. A vaccine, known as EG95, can protect sheep and goats against cystic echinococcosis. This paper describes the adaptation of the EG95 vaccine for use in cattle. The monitoring of results used serology and also infection with E. granulosus eggs, followed by necropsy. Immunisation with living E. granulosus oncospheres showed that cattle could be immunised against E. granulosus. Immunisation of cattle with EG95 plus QuilA was also successful. A dose-response and adjuvant trial showed best results were achieved with 250 μg of antigen and 5mg of the adjuvant QuilA, which was 5 times the recommended sheep dose. After two vaccinations given one month apart, 90% protection was maintained for 12 months. At 12 months a third vaccination boosted protection to 99% which was maintained for a further 11 months.     

Brain metastasis of alveolar echinococcosis in a hyperendemic focus of Echinococcus multilocularis infection

An unusual female case, with alveolar echinococcosis (AE) disseminated from the primary hepatic lesion to the brain by metastasis formation, was retrospectively identified during a community survey in Ningxia Hui Autonomous Region, northwest China in 2003. Among possible metastases of hepatic AE, locations to the brain are rare and usually fatal; and they have especially been assigned to concomitant immune suppression. An enhancing role of pregnancy, which may be suspected in this case, the favourable outcome after surgery and chemotherapy, and also a mental disability in a child following long-term intrauterine exposure to mebendazole, make the report particularly unique.     

Bioinformatics features and immunogenic epitopes of Echinococcus granulosus Myophilin as a promising target for vaccination against cystic echinococcosis

Cystic echinococcosis (CE) is a neglected zoonosis especially in underdeveloped countries around the world. Hence, immunization strategies are beneficial to avert the infection. The present investigation was aimed to predict the primary biochemical characteristics of the EgMyophilin and its potential B-cell and human leukocyte antigen (HLA)-binding epitopes as a promising vaccine candidate. Different web servers were used to predict physico-chemical, antigenic and allergenic profiles, transmembrane domain, subcellular localization, post-translational modification (PTM) sites, secondary and 3D structure, tertiary model refinement and validations. B-cell and HLA-binding epitopes were predicted and screened in terms antigenicity, allergenicity, solubility (B-cell) or hydrophobicity (T-cell). The 89.82 KDa protein was non-allergenic, hydrophilic, stable, with improved thermotolerance and 94 post-translational modification sites. The secondary structure included 42.94% alpha helix, 42.82% random coil and 41.23% extended strand. Based on Ramachandran plot output for refined model, 96.2%, 99.5%, and 0.45% of amino acid residues were incorporated in the favored, allowed, and outlier regions of the refined model, respectively. After epitope screening, four B-cell and five HLA-binding epitopes possessed the highest antigenic index in the protein sequence. This paper is a premise for further researches, and provides insights for the development of a suitable vaccine against CE. More empirical studies are required using the EgMyophilin alone or in combination with other antigens/epitopes in the future.     

Seroprevalence of Echinococcus granulosus infection in a Uruguayan rural human population.

Serological tests using hydatid cyst fluid (HCF) are useful as a primary screen for detecting asymptomatic infections due to Echinococcus granulosus in human populations. An enzyme-linked immunosorbent assay (ELISA) performed with whole sheep HCF was used to determine the prevalence of hydatid antibodies in a rural human population in Uruguay. In order to eliminate cross-reactions with sheep proteins in HCF, 1% normal sheep serum was added to each human serum before testing. ELISA seropositives were further tested using the double diffusion test (DD5) and confirmed where possible by ultrasound and X-ray examination. Serum samples were obtained from 420 individuals inhabiting rural areas in the Department of Paysandu (160), Rivera (21) and San José (239). An overall seroprevalence of 1.24% was obtained. Two of 17 seroreactors were subsequently proven to have hydatidosis. These results confirm and extend previous studies indicating an unusually high prevalence of E. granulosus infection in the Uruguayan population.     

Combined eco-epidemiological and molecular biology approaches to assess Echinococcus granulosus transmission to humans in Mauritania: occurrence of the 'camel' strain and human cystic echinococcosis

Mauritania lies between West-Central Africa where human cystic echinococcosis (CE) is considered extremely rare and West Maghreb where CE accounts for a real public health problem. Until 1992, Mauritania was considered as human CE-free even through CE seemed well known in livestock. In 1992, the introduction of ultrasonography led to the diagnosis of the first human CE cases. In 1997, a veterinary study revealed that dogs living around Nouakchott were commonly infected by Echinococcus granulosus. To assess E. granulosus transmission and to identify the most relevant animal reservoir responsible for human CE emerging in Mauritania, a simultaneous eco-epidemiological and molecular biology approach was performed. The fieldwork included sample collection and investigation of relationship between intermediate hosts, definitive hosts and humans. Typing of E. granulosus strains was performed using comparison of polymerase chain reaction (PCR)-amplified DNA sequences with one nuclear (BG 1/3) and 2 mitochondrial (COI, NDI) targets. Results show that the 'camel' strain is actually infectious to humans and circulates between intermediate hosts including camels and cattle. It is suggested that preventive measures at slaughtering places could reduce human contamination.     

The molecular epidemiology of Echinococcus granulosus and cystic hydatid disease

It is now generally recognized that the dog tapeworm Echinococcus granulosus, the cause of cystic hydatid disease, exhibits substantial genetic diversity. This variability has important implications for the design and development of vaccines, diagnostic reagents and drugs effective against this parasite. The paper describes various deoxyribonucleic acid-based approaches that have been used for accurate identification of these genetic variants and their application in molecular epidemiological surveys of cystic hydatid disease in different geographical settings.     

Epidemiology of Echinococcus granulosus infection in the central Peruvian Andes.

The prevalence of human, canine, and ovine echinococcosis was determined in an endemic area of the Peruvian Andes where control programmes have not been operational since 1980. Prevalence of infection in humans was determined using portable ultrasound, chest X-rays, and an enzyme-linked immunoelectrotransfer blot (EITB) assay. Canine and ovine echinococcal prevalence was determined by microscopic stool examinations following arecoline purging for tapeworm detection and by examination of the viscera from slaughtered livestock animals, respectively. The prevalence among 407 humans surveyed was 9.1%. The frequency of disease in the liver, lung, and in both organs was 3.4%, 2.0%, and 0.2%, respectively. Portable ultrasound or portable chest X-ray has shown that, compared to adults, children under 11 years had significantly higher seropositive rates without evidence of hydatid disease (P < 0.05). Among the 104 dogs inspected for echinococcus after arecoline purging, 33 (32%) were positive for adult tapeworms. Among the 117 sheep slaughtered at the local abattoir, 102 (87%) had hydatid cysts. The prevalence of human hydatidosis in this endemic area of Peru is one of the highest in the world and nearly five times higher than previously reported in 1980. An increase in echinococcosis prevalence may result after premature cessation of control programmes.     

Genetic relationship between the Echinococcus granulosus sensu stricto cysts located in lung and liver of hosts

G1 genotype of Echinococcus granulosus sensu stricto is the major cause of hydatidosis in Northern Africa, Tunisia included. The genetic relationship between lung and liver localization were studied in ovine, bovine and human hydatid cysts in Tunisia. Allozyme variation and single strand conformation polymorphism were used for genetic differentiation. The first cause of genetic differentiation was the host species and the second was the localization (lung or liver). The reticulated genetic relationship between the liver or the lung human isolates and isolates from bovine lung, is indicative of recombination (sexual reproduction) or lateral genetic transfer. The idea of two specialized populations (one for the lung one for the liver) that are more or less successful according to host susceptibility is thus proposed.     

An international study on the serological differential diagnosis of human cystic and alveolar echinococcosis

An enzyme-linked immunosorbent assay (ELISA) for the serological differentiation of cystic (Echinococcus granulosus) and alveolar (E. multilocularis) echinococcosis in man has been evaluated. A discrimination rate of 95.1% was found for 82 sera from patients of geographically disparate endemic areas. This rate was essentially the same as that found for 57 Swiss patients, indicating that inter- and intraspecific strain differences do not influence the test results. The assay method is suitable for immunodiagnostic purposes as well as for seroepidemiological studies.     

Strain identification of Echinococcus granulosus in determining origin of infection in a case of human hydatid disease in Australia

Viable parasitic material from cysts removed surgically from a human patient was used to differentiate the infecting strain of Echinococcus granulosus from another to which the patient may have been exposed. It was shown that the patient was infected with the Australian mainland domestic strain. The age of the cyst material was estimated by microscopical and histological examinations. Combination of these data with an investigation of the patient's life-history allowed identification of the most likely source of infection. Studies such as this help to define those areas of high risk for human hydatidosis on mainland Australia.     

Characterization of Echinococcus granulosus isolates from human, sheep and camel in Iran.

In the present study, Echinoccocus granulosus isolates collected from human, sheep and camel samples in Iran were characterized based on rostellar hook morphology of protoscoleces as well as PCR-RFLP. Morphological study on human and animal isolates showed the presence of two distinct strains of the parasite, one in sheep and the other one in camels. In this regard, rostellar hook of sheep isolates were significantly different from those of camel origin, meanwhile human isolates were found to be similar to those isolated from sheep. Molecular analysis of the ITS1 region of rDNA derived from human, sheep and camel isolates were in agreement with the morphological findings. Based on the PCR-RFLP method, the sheep and human isolates appeared to pertain to the same genotype and the camel isolates were appeared to pertain to a different genotype.     

A field study to control Echinococcus multilocularis-infections of the red fox (Vulpes vulpes) in an endemic focus.

Foxes harbouring E. multilocularis represent an important source for human infection with this parasite which causes alveolar echinococcosis. To minimize the risk of human infection, a control study was conducted to reduce the prevalence of E. multilocularis-infection in foxes in an focal endemic area of 5000 km2. Foxes were given access to baits containing 50 mg praziquantel. Twenty baits per km2 were distributed by airplane during 14 campaigns. The effects of control measures were monitored by parasitological examination of 9387 foxes shot before and during the control trial. A distinct reduction of the prevalence of E. multilocularis was observed for both, the initially endemic area and the low-endemic periphery. The effect was more pronounced in adult than in juvenile foxes. Under control conditions, the risk area decreased in size. However, an eradication of the parasite was not reached with the chosen strategy.     

Spatial distribution patterns of Echinococcus multilocularis (Leuckart 1863) (Cestoda: Cyclophyllidea: Taeniidae) among red foxes in an endemic focus in Brandenburg,Germany.

Over a period of 40 months, 4374 foxes were randomly sampled from an area located in northwestern Brandenburg, Germany, and examined parasitologically for infections with Echinococcus multilocularis. Spatial analysis of the origin of infected animals identified two (one central and one southeastern) high-endemic foci with an estimated prevalence of 23.8%. By contrast, a prevalence of 4.9% was found in the remaining (low-endemic) area. The prevalences among juvenile and adult foxes were compared in the high-endemic and the low-endemic areas. To analyse the central high-endemic focus further, the random sample was stratified by zones representing concentric circles with a radius of 13 km (zone 1) or x(n-1) + 7 km for the remaining three zones from the apparent centre of this focus (anchor point). Prevalences calculated for each zone showed a decrease from zone 1 (18.8%) to zone 4 (2.4%) with significant differences for all zones but zones 3 and 4. The relative risk of an infection decreased rapidly in a distance range of 26 km around the high-endemic focus, whereas the relative risk remained unchanged within a distance of 5 km around the anchor point. The importance of heterogeneous spatial distribution patterns for the diagnosis and epidemiology of the infection is discussed.     

Immunodiagnostic differentiation of alveolar and cystic echinococcosis using ELISA test with 18-kDa antigen extracted from Echinococcus protoscoleces

Immunoblotting analysis was carried out using crude extracts of Echinococccus multilocularis and E. granulosus. A total of 214 serum samples were examined for the detection of antibody responses against 18-kDa polypeptides (Eg18 and Em18). Of 44 sera from patients with alveolar echinococcosis (AE) 91% were positive against both Eg18 and Em18, compared with 10% and 13%, respectively, of 70 cystic echinococcosis (CE) samples and 13% and 17% of 29 cysticercosis samples. A relatively purified 18-kDa antigen was extracted from E. granulosus protoscoleces and an 18-kDa-ELISA test which is simple, fast and highly sensitive and specific has been established. A comparative ELISA analysis, using the purified 18-kDa antigen and pooled AE and CE serum, indicated that there are significant differences in antibody levels between AE and CE sera. The maximum ratio for the evaluation of AE to CE was 2.26. The 18-kDa-ELISA was positive for 91% (40/44) and 11% (8/70) of AE and CE sera, respectively, and there was no cross-reactivity with cysticercosis sera or healthy controls. The overall diagnostic values of 18-kDa-ELISA for AE sera were sensitivity 91%, specificity 94%, positive predictive value 83% and negative predictive value 97%. We conclude that (i) the 18-kDa antigenic component may not be species specific but (ii) levels of antibody to the 18-kDa antigen are significantly different between AE and CE sera. Therefore, the 18-kDa antigen can be a reliable serological marker for differentiation of AE from CE.     

Several strains of Echinococcus granulosus infect livestock and humans in Argentina.

Mitochondrial cytochrome oxidase subunit 1 (CO1) sequencing, Southern blot of a repetitive DNA element and single strand conformation polymorphism of the 5' non-transcribed region of the cytosolic malate dehydrogenase (MDH) gene were used to determine the extent and distribution of Echinococcus granulosus genetic variation in Argentina. Five distinct strains of E. granulosus were shown to exist in the country. The common sheep, Tasmanian sheep, cattle and camel strains were identified in humans. Unlike the situation found in other countries, where the common sheep strain is the major source of human contamination, the Tasmanian sheep and camel strains produced a significant number of human infections in some regions of Argentina. This is the first report of cattle strain in humans in South America. Goats could be the natural intermediate host of the camel strain, which was not identified in humans from other regions so far. More than one genotype was identified in the same geographic area. These findings may have important consequences for human health and the control of hydatid disease. Within-strain differences were also observed, showing the potential of variation of E. granulosus.     

Changing prevalence of hepatitis C virus infection among teenagers in an endemic area in Taiwan

Tzukuan Township in Taiwan has been reported to be an endemic area for hepatitis C virus (HCV) infection both in adults and adolescents. The maritime part of the township carries a higher prevalence than the non-maritime part and, as a consequence, several public education strategies have been introduced during the past decade. The current follow-up study aimed to clarify the changing prevalence of HCV infection among teenagers in the endemic maritime part of Tzukuan. In addition to viral hepatitis markers and biochemical profiles, we compared the epidemiological characteristics of 887 and 394 teenagers (aged 13-16 years) from the maritime part enrolled in 1995 and 2005, respectively. Compared with the results of surveillance in 1995, the prevalence of anti-HCV seropositivity (1.0% vs. 2.8%; P=0.045) and HCV RNA (0.5% vs. 2.3%; P=0.026) had decreased significantly by 2005. Transfusions and anti-HCV-positive families were the main risk factors amongst the 25 anti-HCV-positive teenagers in 1995, and became non-significant amongst the four anti-HCV-positive teenagers in 2005. In conclusion, the seroprevalence of HCV infection has significantly decreased after one decade of intervention among the teenage population in this endemic area.     

Multicentre clinical trials of benzimidazole-carbamates in human cystic echinococcosis (phase 2)

A multicentre study which constituted the second phase of trials of the efficacy of albendazole and mebendazole in human cystic echinococcosis was coordinated by WHO. A total of 112 patients from four clinical centres in Beirut, Paris, Rome and Sofia completed standardized dosage of regimens of each drug and 68 patients were followed up for at least 12 months after treatment. Albendazole was more effective than mebendazole and adverse reactions were comparable with both treatment regimens. At least 12 months is needed after treatment for an objective evaluation of the efficacy of benzimidazoles. At present, treatment with albendazole or mebendazole should be reserved for inoperable cases of cystic echinococcosis (under strict medical supervision) and individualized according to the patient's response and the occurrence and severity of adverse reactions.     

Field trials of various molluscicides (chiefly sodium pentachlorophenate) for the control of aquatic intermediate hosts of human bilharziasis

Field trials of various molluscicides-notably sodium pentachlorophenate, copper sulfate and dinitro-o-cyclohexylphenol-were carried out in Brazil in order to test their effectiveness against the aquatic intermediate hosts of human bilharziasis. Sodium pentachlorophenate was found to be the most effective in relation to cost and availability. Subsequent tests in Egypt, using sodium pentachlorophenate alone, while confirming the effectiveness of this molluscicide in bilharziasis control, have shown that the dosage and methods of application must be adapted to environmental conditions.     

Assessment of the immunological surveillance value of humoral and lymphocyte assays in severe human cystic echinococcosis

Thirty cystic echinococcosis (CE) patients in Uruguay with severe bone or secondary disseminated echinococcosis were immunologically assessed using cellular (lymphocyte transformation assay, LTA) and humoral (specific antibody and subclass responses, circulating antigen and circulating immune-complexes) immunological assays during the course of chemotherapy (albendazole and/or praziquantel). CE patients were divided into 4 groups, according to clinical treatment and outcome: (I) surgery and chemotherapy, (II) chemotherapy with outcome unchanged, (III) chemotherapy with outcome improved, and (IV) chemotherapy considered cured. Increased circulating antigen was of prognostic value in some severe CE cases where levels remained high and/or increased. The lymphoproliferative response in vitro to Echinococcus granulosus antigen was statistically greater in all patient groups compared to normal individuals but at lower levels in improved or cured CE patients. Levels of non-specific LTA response were significantly lower than controls for all groups during albendazole treatment (P < 0.001) but returned to normal levels in cured patients, a result consistent with parasite-induced suppression of cellular responses. This study suggests that, at least in severe osseous and secondary CE, immunosurveillance by specific antibodies, especially total specific immunoglobulin, was overall of more practical use than antigen-specific in-vitro lymphocyte transformation assays.