Transcriptome analysis of Taenia solium cysticerci using Open Reading Frame ESTs (ORESTES)

Background: Human infection by the pork tapeworm Taenia solium affects more than 50 million people worldwide, particularly in underdeveloped and developing countries. Cysticercosis which arises from larval encystation can be life threatening and difficult to treat. Here, we investigate for the first time the transcriptome of the clinically relevant cysticerci larval form. Results: Using Expressed Sequence Tags (ESTs) produced by the ORESTES method, a total of 1,520 high quality ESTs were generated from 20 ORESTES cDNA mini-libraries and its analysis revealed fragments of genes with promising applications including 51 ESTs matching antigens previously described in other species, as well as 113 sequences representing proteins with potential extracellular localization, with obvious applications for immune-diagnosis or vaccine development. Conclusion: The set of sequences described here will contribute to deciphering the expression profile of this important parasite and will be informative for the genome assembly and annotation, as well as for studies of intra- and inter-specific sequence variability. Genes of interest for developing new diagnostic and therapeutic tools are described and discussed.

Study on some biological and feeding factors of spined loach Cobitis taenia from the Babolrud, Talar and Siahrud Rivers

Spined loach, Cobitis taenia, is a predominant fish in the river systems of the southern Caspian Sea basin. Although there is evidence of the geographical divergence of this taxon, but no information is available on morphological differences within the species populations. This study was designed to evaluate some biological factors including; morphometric and meristic characters, length-weight, age-growth, condition factor, diet, reproduction, variation and differentiation, in the Babolrud, Talar and Siahrud Rivers in south of the Caspian Sea basin. Age, sex ratio, fecundity, ova diameter and gonadosomatic index were estimated. Also, regression analyses was tested the relation between fecundity and fish length, weight, gonad weight, and also age. Totally 858 fish of which 721 were matures, were collected from these rivers by electrofishing. 37 morphometric characters, 9 meristic characters and 78 truss network system characters were estimated. Resulats of DFA analysis based on data of morphometric and meristic showd that these populations are highly (94.5%) varios from each other. In discriminate function analysis, the proportion of individuals correctly classified into their original groups was 61%, 65.4% and 86.5% for upstream and downstream, respectively. Clustering based on Euclidean distances among groups of centroids using an UPGMA and also principal component analysis’ results for morphometric data indicated that these populations from these three rivers were clearly distinct from each other. Regression equations between length and weight in these three populations were significantly different from Folton factor (b=3), that showed the fish has a negative Alometric growth process. Condition factor was estimated between 0.8912 to 1.2736 and 0.8131 to 1.4489 for males and females, respectively. Sex ratio (female: male) in these populations was 1.2816:1. The difference between the number of females and males was significant and females were more than males. The female and male specimens reach maturity by Tl more than 40 and 30 mm and at the age of 2+ and 1+, respectively. The mean of ova diameter was 0.5824±0.2882. The spawning took place from May to late July, at the water temperature from 18.7 to 24.0°C. The GSI values average at the beginning of the reproduction period was about 9%, with ranged from 2 to 26% in ripe mature females. The absolute and relative fecundity were 2109±792 and 579±208 respectively. The absolute fecundity was significantly related to body weight and gonads weight. Based on the pattern of gonado-somatic index, it was concluded that this fish has prolong active reproductive period, which is a type of adaptation by short-lived small fishes to environmental conditions. The macroscopic and histological results showed that the female and male have 5 and 4 stages in their maturation process, respectively. The RLG index was about 0.4732, which showed the fish is a carnivorous species. Significant difference was observed between fishes with different length and diet. The main foods of the fish were Trichoptera, Chironomidae larvae and Ephemeroptera which were their prefered food as well, however it was estimated that the food selection and diet are affected by environmental conditions.

Identification and characterization of a cathepsin L-like cysteine protease from Taenia solium metacestode

Taenia solium metacestode, a larval pork tapeworm, is a causative agent of neurocysticercosis, one of the most common parasitic diseases in the human central nervous system. In this study, we identified a cDNA encoding for a cathepsin L-like cysteine protease from the T solium metacestode (TsCL-1) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1216 bp encoded 339 amino acids with an approximate molecular weight of 37.6 kDa which containing a typical signal peptide sequence (17 amino acids), a pro-domain (106 amino acids), and a mature domain (216 amino acids). Sequence alignments of TsCL-1 showed low sequence similarity of 27.3-44.6 to cathepsin L-like cysteine proteases from other helminth parasites, but the similarity was increased to 35.9-55.0 when compared to mature domains. The bacterially expressed recombinant protein (rTsCL-1) did not show enzyme activity; however, the rTsCL-1 expressed in Pichia pastoris showed typical biochemical characteristics of cysteine proteases. It degraded human immunoglobulin G (IgG) and bovine serum albumin (BSA), but not collagen. Western blot analysis of the rTsCL-1 showed antigenicity against the sera from patients with cysticercosis, sparganosis or fascioliasis, but weak or no antigenicity against the sera from patients with paragonimiasis or clonorchiasis. (c) 2006 Published by Elsevier B.V.

Taenia solium transmission in a rural community in Honduras : an examination of risk factors and knowledge

Taenia soliurn taeniasis and cysticercosis are recognized as a major public health problem in Latin America. T. soliurn transmission not only affects the health of the individual, but also social and economic development, perpetuating the cycle of poverty. To determine prevalence rates, population knowledge and risk factors associated with transmission, an epidemiological study was undertaken in the rural community of Jalaca. Two standardized questionnaires were used to collect epidemiological and T. soli urn general knowledge data. Kato-Katz technique and an immunoblot assay (EITB) were used to determine taeniasis and seroprevalence, respectively. In total, 139 individuals belonging to 56 households participated in the study. Household characteristics were consistent with conditions of poverty of rural Honduras: 21.4% had no toilet or latrines, 19.6% had earthen floor, and 51.8% lacked indoor tap water. Pigs were raised in 46.4% of households, of which 70% allowed their pigs roaming freely. A human seroprevalence rate of 18.7% and a taeniasis prevalence rate of 2.4% were found. Only four persons answered correctly 2: 6 out of ten T. soliurn knowledge questions, for an average passing score of 2.9%. In general, a serious gap exists in knowledge regarding how humans acquire the infections, especially neurocysticercosis was identified. After regression analysis, the ability to recognize adult tapeworms and awareness of the clinical importance of taeniasis, were found to be significant risk factors for T. soliurn seropositivity. These results demonstrate a high level of transmission and a low level of kn~,wledge about Taenia soliurn in Jalaca. Consequently, intervention measures integrated with health education are necessary to decrease the burden caused by this parasite.

Detection of IgG in cerebrospinal fluid for diagnosis of neurocysticercosis: evaluation of saline and SDS extracts from Taenia solium and Taenia crassiceps metacestodes by ELISA and immunoblot assay

We compared saline (S) and sodium dodecyl sulphate (SDS) extracts from Taenia solium (homologous species - HO) and Taenia crassiceps (heterologous species - HE) metacestodes in order to detect Ige by ELISA and immunoblot assay (IBA) in cerebrospinal fluid (CSF) for the diagnosis of human neurocysticercosis (NC). CSF samples were obtained from 93 patients. of these, 40 had NC, five had a diagnosis of probable NC, nine had central nervous system schistosomiasis or strongyloidiasis and 39 had other neurological alterations. Samples were analysed by ELISA and the results were compared with IBA in all samples with confirmed and probable NC diagnosis, in all samples with other central nervous system parasitic infection, and in 10 of those with another neurological alterations. ELISA sensitivity was 100%, 85%, 95% and 87.5% for the S-HO, S-HE, SDS-HO and SDS-HE extracts, respectively, and ELISA specificity was 100% for S-HO, S-HE, SDS-HO extracts and 97.9% for SDS-HE antigen. Immunodominant peptides detected by IBA were, by decreasing percentage of recognition: 64-68 and 45 kDa for S-HO; 108-114, 92-95, 64-68, 83 and 88 kDa for S-HE; 64-68, 108-114, 77 and 86 kDa for SDS-HO; and 108-114, 88 and 92-95 kDa for SDS-HE. Overall the homologous antigenic extracts showed higher sensitivity than the heterologous extracts in the diagnosis of NC in CSF samples. The heterologous extracts contained most of the immunodominant peptides presented in the homologous extracts, which are recognized by Ige antibodies in CSF samples.

Taenia solium DNA is present in the cerebrospinal fluid of neurocysticercosis patients and can be used for diagnosis

Neurocysticercosis is the most frequent parasitic infection of the CNS and the main cause of acquired epilepsy worldwide. Seizures are the most common symptoms of the disease, together with headache, involuntary movements, psychosis and a global mental deterioration. Absolute diagnostic criteria include the identification of cysticerci, with scolex, in the brain by MRI imaging. We demonstrate here, for the first time, that T. solium DNA is present in the cerebrospinal fluid of patients. The PCR amplification of the parasite DNA in the CSF enabled the correct identification of 29/30 cases (96.7 %). The PCR diagnosis of parasite DNA in the CSF may be a strong support for the diagnosis of neurocysticercosis.

Comparative evaluation of an indirect ELISA test for diagnosis of swine cysticercosis employing antigen from Taenia solium and Taenia crassiceps metacestodes

Taenia solium cysticercosis is still a serious public health problem in several countries where poverty and lack of hygiene favor transmission. Because pigs are the primary intermediate hosts, prevalence of porcine cysticercosis is a reliable indicator of active transmission zones. Serological diagnostic methods are important tools for epidemiological studies since they can be applied to living animals on a large scale. Four antigen preparations (cyst fluid and crude) from T. solium and T. crassiceps metacestodes were compared for swine cysticercosis diagnosis by indirect ELISA (IE). Twenty-eight serum samples from swine naturally and experimentally infected by cysticerci of T. solium and 56 serum samples from swine reared in commercial herds were tested. Best results of overall sensitivity were obtained by the use of cyst fluid and crude antigen of T. crassiceps metacestode (100 and 96.4%, respectively). Using homologous antigen preparations we have observed higher specificity percentage (98.2% for cyst fluid and 96.4% for crude metacestode T. solium antigen). We concluded that sensitivity is of far more importance than specificity for identification of endemic areas in order to prevent transmission to man. We conclude, therefore, that IE performed with cyst fluid antigen of T. crassiceps metacestode is a better tool for that purpose. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Immunodiagnosis of swine cysticercosis by indirect ELISA employing a heterologous antigen from Taenia crassiceps metacestode

C. Larralde et al. (1990, Arch. Pathol. Lab. Med., 114:926-928) demonstrated that heterologous antigen from the laboratory-adapted murine Taenia crassiceps metacestode may substitute those from Taenia solium in the immunodiagnosis of human cysticercosis by the indirect enzyme-linked immunosorbent assay (IE). ?This antigen is easily obtained at a laboratory level and solves the problem of T. solium cysticerci collection from naturally or experimentally infected swine. In this study an IE employing a heterologous antigen from the T. crassiceps metacestode was evaluated for the immunodiagnosis of swine cysticercosis. Sera from 300 swine free of T. solium cysticerci by post-mortem examination were employed to determine two IE cut- off values: 1) Mean ELISA values + 2 standard deviations (2 sigma cut-off) and 2) - Mean ELISA values + 3 standard deviations (3 sigma cut-off). The specificity of IE was 97% with the 2 sigma cut-off and 100% with the 3 sigma cut-off. When applied to ten sera from swine infected by cysticerci of T. solium by post-mortem examination, the sensitivity of IE was 100% independent of the cut off.

Investigação epidemiológica em plantio de alface (Lactuca sativa) na cidade de Botucatu - SP visando detectar a presença de ovos de Taenia sp

Pós-graduação em Medicina Veterinária - FMVZ

Cysticercosis in experimentally and naturally infected pigs: Parasitological and immunological diagnosis

Silva M. R. M., Uyhara C.N.S., Silva F. H., Espindola N.M., Poleti M. D., Vaz A.J., Meirelles F. V. & Maia A. A. M. 2012. Cysticercosis in experimentally and naturally infected pigs: Parasitological and immunological diagnosis. Pesquisa Veterinaria Brasileira 32(4): 297-302. Departamento de Ciencias Basicas, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de Sao Paulo, Av. Duque de Caxias Norte 225, Pirassununga, SP 13635-900, Brazil. E-mail: antomaia@usp.br. Our objective was to evaluate the diagnosis of swine cysticercosis by examining "ante mortem" (inspection of the tongue), "post mortem" (inspection and detailed necropsy) and ELISA for research in serum of antibodies (Ab-ELISA) and antigens (Ag-ELISA). Seven (7) pigs were experimentally infected orally with eggs of Taenia solium and another 10 were naturally infected. In the pigs experimentally infected, inspection of the tongue was negative in all animals, in the routine inspection detailed necropsy and cysticercis were identified in all of them. In pigs with heavy natural infection, inspection of the tongue identified cysticerci in two (20%), while at inspection with necropsy the parasites were identified in large quantities in all animals. In ELISA for antibody search (Ab-ELISA) TS-14 recombinant protein was used, and in search for antigen (Ag-ELISA) a monoclonal antibody against this protein. In animals experimentally infected, blood was collected weekly for 140 days. The Ab-ELISA identified an increase in titers of antibody to cysticerci 21 days after infection, and at the end of the experimental period six animals (86%) were positive to the test. The search for circulating antigens (Ag-ELISA) was positive in two pigs 28 to 91 days after infection. All naturally infected pigs were positive for Ag-ELISA and Ab-ELISA. The search for antibodies and antigens by ELISA in serum from 30 pigs of a local farm and without history of cysticercosis was negative. Thus, the use of TS-14 antigen in ELISA test (Ab-ELISA) can be useful for the diagnosis of cysticercosis in pigs with low infection.

Demonstration of species-specific and cross-reactive components of Taenia solium metacestode antigens.

Analysis of human serum reactivities to antigenic components of soluble Taenia solium metacestode proteins showed the predominant presence of determinants shared by T. solium, Echinococcus multilocularis and E. granulosus. Two polypeptides were demonstrated by SDS-PAGE and Western blot or enzyme-linked immunoelectrotransfer blot (EITB) assay to bind serum and CSF antibodies only from T. solium cysticercosis patients. These species-specific antigenic polypeptides focused between pH 4.6 and 3.9 after resolution by isoelectric focusing followed by EITB. The high species-specificity demonstrated by the present techniques offers the opportunity to confirm serologically an infection by T. solium metacestode.

Dissertation sur le taenia ou ver-plat, dans lequelle on prouve que ce ver n'est pas solitaire : avec une lettre sur la poudre de sympathie, propre contre le rhumatisme simple ou gouteux : On y a joint la manière de l'apprêter & de s'en servir & le discours prononcé par M. ... Digby sur l'efficacité de cette poudre /

Mode of access: Internet.

Encroachment of Echinococcus granulosus into urban areas in eastern Queensland, Australia

Objective: To investigate the prevalence of Echinococcus granulosus in wild dogs (dingos and dingo-domestic dog hybrids) living in and around human habitation on Fraser Island and in townships of the Maroochy Shire, on Queensland's Sunshine Coast, Australia. Design: Wild dogs were humanely killed on Fraser Island and in the Maroochy Shire because they were deemed a potential danger to the public. Their intestines were collected and the contents examined for intestinal parasites. Procedure: Intestines were removed as soon after death as possible, packed in plastic bags and kept frozen until examination. The intestinal contents were washed, sieved and examined microscopically for the presence of helminths, which were identified and counted. Results: Intestines from 108 wild dogs, 7 foxes and 18 Fraser Island dingoes were examined. Echinococcus granulosus was only present in the wild dogs from Maroochy Shire (46.3%) with worm burdens of between 30 and 104,000. Other helminths included Spirometra erinacei, Dipylidium caninum, Taenia spp., Ancylostoma caninum and Toxocara canis. Two specimens of a trematode (Haplorchinae sp.) usually found infecting fish and seabirds were recovered from a Fraser Island dingo. Conclusion: Dingoes on Fraser Island are not infected with E. granulosus and do not pose a hydatid disease public health risk to residents or visitors. However, wild dogs examined from the Maroochy Shire do present a potential hydatid disease public health risk.