Serodiagnosis of sheep hydatidosis with hydatid fluid, protoscolex, and whole body of Echinococcus granulosus antigens

Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. A great number of immunological assays have been developed for detection of anti-hydatid cyst antibodies. The principal intermediate host of Echinococcus granulosus in most endemic regions of the world is sheep. Antibodies to various antigens are detectable in the sera of some, but not all infected sheep. The objective of the present study was to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis in sheep with different (hydatid fluid, protoscolices, and whole body of E. granulosus) antigens. A total of 100 sera were collected from sheep with hydatidosis proven by inspection of hydatid-infested livers and lungs of the sheep slaughtered in Mashhad abattoir. Hydatid fluid and protoscolex were isolated from livers or lungs of sheep with hydatid cyst in sterile conditions. Whole body of E. granulosus was isolated from intestine of infected dogs. Sera samples were examined by ELISA with different antigens. The results of antibody detection by indirect ELISA, using different antigens, showed that the hydatid fluid was the most effective antigen of those assessed for detection of infection with hydatidosis in sheep. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst in sheep.     

The phylogenetic similarity of hydatid cyst isolated from humans and sheep in Ilam Province southwest of Iran

Hydatid cyst is a chronic zoonotic disease caused by the larval stage of the dog tapeworm, Echinococcus granulosus. To identify genotype of hydatid cysts of human and sheep jackal in Ilam Province (South West of Iran), the PCR-RFLP and DNA sequencing were used. A total of 10 human and 20 sheep protoscoleces hydatid cyst samples were collected from different hospitals and slaughterhouses. Then, the gene of cox1 of mitDNA of the parasite was amplified and PCR products were cut using AluI and HpaII restriction enzymes. Finally, a number of PCR products were bi-directionally sequenced. Based on the DNA sequencing and PCR-RFLP results, human and sheep samples indicated to pertain the genotypic similarities. Our data indicated that, the genotypes of larval stage of E. granulosus is similar in both intermediate hosts. According to the phylogenetic tree, there is at least one genotype of parasite, which belongs to E. granulosous sensu stricto (G1–G3) complex and overall isolates sequences of mtDNA indicated 100 % homology with references G1, G2, and G3 sequences in the GenBank database. G1 genotype was the dominant genotype of human and livestock.     

Prevalence of hydatidosis in slaughtered sheep and goats by season,sex, age,and infected organ at Amol abattoir,Mazandaran Province,Iran

Hydatidosis is one of the zoonotic diseases with special importance for public health and causing financial problems in developed and developing countries. This infection is the most prevalent disease in the Middle East especially in Iran. In this survey, internal organs of 1,200 sheep and 1,200 goats, including liver, lung, heart, and kidney, were randomly inspected to estimate prevalence rate of hydatidosis and its relationship with season, sex, age, and infected organs. It was found that a total of 335 (27.9 %) of the 1,200 slaughtered goats and a total of 546 (45.5 %) of the 1,200 slaughtered sheep were infected with the hydatid cysts. Prevalence of hydatidosis in females was significantly (p?<?0.05) more than males, and the infection rate was significantly (p?<?0.05) increased by age. The most infected organs were liver and lung, respectively, and the least infected organs were kidney and heart, respectively. High prevalence of hydatidosis in Iran can be a result of conventional slaughtering of sheep and goats, availability of carcass wastes and offal for scavenging stray dogs and other wild carnivores, and close relation between shepherd dog and these animals. The prevalence rate can be decreased by interrupting Echinococcus granulosus life cycle, stopping illegal slaughtering, and increasing public awareness about the infection.     

Prevalence of human papillomavirus infection in Argentinean women attending two different hospitals prior to the implementation of the National Vaccination Program

Cervarix vaccine was included in the National Immunization Program of Argentina in 2011 but data about the local distribution of human papillomavirus (HPV) infection in women exposed to the virus are scarce. This cross‐sectional study determined the prevalence and type distribution of HPV infection in unvaccinated women attending routine gynecological screening in two public hospitals located in Buenos Aires and Santa Fe, Argentina. Socio‐demographic, sexual behavior, and co‐factors information was obtained from all participants (Buenos Aires, n = 429; Santa Fe, n = 433). Cervicovaginal swabs were tested with an MY11/09 primer‐based assay and with the CUT primer system targeting mucosal/cutaneous HPVs. Participants from Buenos Aires showed significantly higher rates of HPV infection (52.4% vs. 40.6%), of multiple infections (24.2% vs. 16.4%), and of low‐risk (20.3% vs. 13.9%) and high‐risk types (44.1% vs. 33.3%) than those from Santa Fe. HPV‐66 (Buenos Aires: 17%) and HPV‐16 (Santa Fe: 8.5%) were the most prevalent types. Novel HPV‐66 putative subtype and variants were identified. Vaccine types 16 and 18 were frequent (Buenos Aires: 13.5%; Santa Fe: 10.2%) but few participants had co‐infections with both (Buenos Aires: 1.4%; Santa Fe: 0.2%). A common risk factor for HPV infection was having a new sexual partner in the last year (Buenos Aires: OR 2.53, P < 0.001; Santa Fe: OR 1.85, P = 0.04). This study provides valuable baseline data for future assessment of the impact of massive vaccination in Argentina and it underlines the use of additional HPV testing strategies, such as the CUT system, for surveillance and vaccinology. J. Med. Virol. 85:655–666, 2013. © 2013 Wiley Periodicals, Inc.     

Production of IL-5 and IL-6 by peripheral blood mononuclear cells (PBMC) from patients with Echinococcus granulosus infection

The role of Th2 cytokines in human hydatidosis was evaluated in ELISA determining IL-5 and IL-6 production in PBMC cultures from 27 pharmacologically treated hydatid patients and from 13 uninfected controls. PBMC from patients produced large amounts of parasite antigen-driven IL-5, whereas PBMC from uninfected individuals produced none. In contrast, PBMC from patients and from uninfected controls produced large amounts of parasite antigen-driven IL-6. Immunoglobulin isotype analysis revealed that IL-5 production correlated significantly with IgE and IgG4 expression (IL-5/IgE r = 0.5, P<0.05; IL-5/IgG4 r = 0.6, P<0.05). The high IL-5 levels in supernatants from patients’ PBMC did not correspond to an increase in eosinophils. Neither IL-5 nor IL-6 production showed an association with the outcome of therapy. Overall, these findings confirm that the lymphocytes of individuals with Echinococcus granulosus infection contain Th2-like subpopulations.     

Molecular characterization of Echinococcus granulosus in a hyperendemic European focus,the Republic of Moldova

Cystic echinococcosis is a zoonosis caused by the tapeworm Echinococcus granulosus sensu lato. The lifecycle of the parasite is mainly domestic, requiring dogs as definitive hosts and livestock species as intermediate hosts. Although human cystic echinococcosis is a high public health priority in the Republic of Moldova, the rare animal data available concerns only infection in cattle. A preliminary slaughterhouse survey was conducted to assess prevalence and perform the first molecular characterization of E. granulosus sensu lato in sheep and cattle. For the survey, 40 sheep and 19 cattle were inspected. Very high prevalence in sheep (82.5 %) and in cattle (78.9 %) was found. Molecular analyses identified genotypes G1 and G3 of E. granulosus sensu stricto in all the liver and lung samples. Based on the concatenated sequences of cox1?+?nad3 (701 bp), 23 different haplotypes were obtained. Mixed infections by different haplotypes/genotypes were frequently identified in both sheep and cattle. The relatively high (20.0 %) cyst fertility observed in cattle argues for the potential contribution of cattle to the lifecycle of E. granulosus sensu stricto, unlike previous observations in Europe. The hyperendemic situation of Moldova can be explained by a high majority of animals slaughtered at home usually without veterinary inspection. Further extensive slaughterhouse surveys with molecular identification also involving pigs and goats are needed to obtain a better overview of the epidemiological situation of E. granulosus sensu lato in this hyperendemic focus in the Republic of Moldova.     

An immunoblot for detection of Taenia saginata cysticercosis

Control measures to prevent human infections with the food-borne zoonotic helminth Taenia saginata are currently based on meat inspection, which shows rather low diagnostic sensitivity. To develop an immunoblot for detection of T. saginata-infected cattle, crude proteins of T. saginata cysts were extracted and separated with SDS-PAGE. The cyst antigens showed ten protein bands ranging from 260 to 14 kDa. T. saginata cyst proteins 260, 150, 130, 67, 60, 55, 50, and 23 kDa were immunoreactive with known positive sera of T. saginata-infected cattle but cross-reacted with sera from Echinocccus granulosus-infected ruminants. By contrast, 14- and 18-kDa cyst proteins reacted specifically with T. saginata-positive sera and thus might be potential candidates for the development of a T. saginata-specific immunoassay. Proteins of E. granulosus cysts and Taenia hydatigena cysts were also extracted and separated with SDS-PAGE. E. granulosus cysts revealed 11 protein bands ranging from 260 to 23 kDa. E. granulosus protein 60 kDa was immunoreactive with E. granulosus-positive sera only. The cyst of T. hydatigena showed 11 protein bands ranging from 290 to 14 kDa. The protein band 35 kDa showed cross-reaction with positive sera from both T. saginata- and E. granulosus-infected animals. A protein of 67 kDa was present in all three tested cestode species and was the major antigenic protein detected by sera of T. saginata- and E. granulosus-infected animals. Therefore, this protein represents a potential vaccine candidate against both cysticercosis and cystic echinococcosis in cattle.     

Serodiagnosis and seroepidemiology of human unilocular hydatidosis in Jordan

A total of 2182 serum samples from 38 patients with surgically confirmed unilocular hydatidosis, 19 clinically assessed patients, 15 patients with parasitic infections other than hydatidosis, 104 hospital outpatients, and 2006 normal Jordanians were serodiagnosed for the presence of IgG antibodies against hydatid fluid, circulating immune complexes (CIC), and/or hydatid circulating antigen (CA). Anti-hydatid IgG antibodies were detected in the sera of 77.4% of patients with hydatid discase and persiste for very long periods postsurgery. As many as 54.1% of patients with hydatidosis had positive levels of CIC, and 16.1% had circulating antigen in their sera. The search for circulating antigen and CIC decreased the number of false-negative hydatid cases from seven to three, and the combined sensitivity of the assays thus increased from 77.4% to 90.3%. Using the immunoblot technique, 16- and <14.4-kDaEchinococcus granulosus-specific bands were detected in sera from 54.1% and 61.5% of patients with hydatid disease who were tested before and after surgery, respectively. The seropositivity rate for anti-hydatid IgG antibodies was 2.4% for the general Jordanian population and 5.8% for hospital outpatients.     

Cystic Echinococcosis

Echinococcosis is one of the 17 neglected tropical diseases (NTDs) recognized by the World Health Organization. The two major species of medical importance are Echinococcus granulosus and Echinococcus multilocularis. E. granulosus affects over 1 million people and is responsible for over $3 billion in expenses every year. In this minireview, we discuss aspects of the epidemiology, clinical manifestations, and diagnosis of cystic echinococcosis or cystic hydatid disease caused by E. granulosus.     

Serological Diagnosis of Lung Cystic Hydatid Disease Using the Synthetic p176 Peptide

Cystic hydatid disease (CHD) is a worldwide zoonosis caused by the larval stage of the dog tapeworm Echinococcus granulosus. Diagnosis is based on imagenological tools (abdominal ultrasound, chest X-rays, or computed tomography [CT] scan). Serological antibody-detecting assays, using diverse native antigens, have been used as a supportive diagnostic tool, but their sensitivities and specificities differ greatly. The use of synthetic peptides as antigens should provide more reliability and allow better assessment and comparison of test formats and case series. The synthetic peptide p176, corresponding to the N-terminal extreme of the subunit of antigen B (AgB8/1), has shown promising performances for diagnosis of CHD. We evaluated the performance of the synthetic peptide p176 for the diagnosis of pulmonary hydatid disease in an enzyme-linked immunosorbent assay (ELISA) format. Sixty-one serum samples from patients with a diagnosis of pulmonary hydatidosis confirmed by surgery and 128 from healthy volunteers were tested. The overall sensitivity and specificity of the p176 ELISA for lung CHD were 78.69% and 96.88%, respectively. On bivariate analysis, positive serum antibody reactions were associated with the presence of complications and with the number of cysts (single/multiple). Only the presence of persistent complications significantly associated with seropositivity on multivariate logistic regression analysis (odds ratio [OR], 9.58; 95% confidence interval [CI], 2.15 to 42.6; P = 0.003). The p176 ELISA performs well for the diagnosis of lung CHD and adds an easily reproducible diagnostic assay to the existing diagnostic tools.     

Copro-DNA test for diagnosis of canine echinococcosis

Echinococcosis is one of the most important parasitic zoonotic diseases in the world. The life cycle of Echinococcus granulosus is dependent to the dog–sheep cycle and is actively transmitted in all pastoral regions where sheep, cattle and camels predominate. DNA approaches are now being used routinely for accurate identification of Echinococcus and Taenia species, subspecies and strains. In this study, faecal samples were collected from 50 stray dogs from Mashhad city in the northeast of Iran during June 2011. All samples were frozen at least for 1 week in ?80°C. The embryophore layer of the eggs was broken by freezing–thawing method after egg concentration by the formalin-ether method. DNA was extracted using a DNA isolation kit (MBST, Germany/Iran) according to the manufacturer’s instructions. After PCR, by the primers expressed in the following it became, clear that about 20 % of stray dogs are infected with E. granulosus. In this study, we describe a modified method for DNA isolation from faeces for coprodiagnosis of Echinococcus spp.     

Prevalence survey and first molecular characterization of Echinococcus granulosus in France

Human cystic echinococcosis (hydatid disease) caused by the Echinococcus granulosus tapeworm continues to be a substantial cause of morbidity and mortality in many parts of the world. France is still considered as endemic area, but the current infestation by E. granulosus of intermediate hosts in France remains currently unknown due to the absence of official data reporting for the last 20 years. A 1-year prevalence survey was conducted in the 24 slaughterhouses of ten departments of the South of France. We demonstrate that the E. granulosus parasite is still currently present at low prevalence at slaughterhouses in the study area (4 cases for 100,000 sheep and 3 cases for 100,000 cattle). In addition, we assess the presence of genotype G1 in infected animals and identify for the first time in France genotypes G2 and G3 of E. granulosus sensu stricto.     

Diethylaminoethyl (DEAE) binding fraction from Taenia solium metacestode improves the neurocysticercosis serodiagnosis

Neurocysticercosis (NC) is one of the most important diseases caused by parasites affecting the central nervous system. We fractionated by ion-exchange chromatography using diethylaminoethyl (DEAE)-sepharose resin the total saline extract (S) from Taenia solium metacestodes and evaluated obtained fractions (DEAE S1 and DEAE S2) by enzyme-linked immunosorbent assay (ELISA, n?=?123) and immunoblotting (IB, n?=?22) to detect human NC in serum. Diagnostic parameters were established by ROC and TG ROC curves for ELISA tests. IB was qualitatively analyzed. S and DEAE S1 presented sensitivity of 87. 5 % and DEAE S2 90 %. The best specificity was observed for DEAE S2 (90.4 %). In IB, using DEAE S2 samples from NC patients presented bands of 20–25, 43–45, 55–50, 60–66, 82, 89, and 140 kDa. The great diagnostic parameters reached by DEAE S2 suggest the potential applicability of this fraction in NC immunodiagnosis.     

Unusual locations of hydatid disease: A 10-year experience from a tertiary reference center in Western Turkey

IntroductionHydatid disease is an endemic parasitic infection caused by Echinococcus granulosus mostly seen in the Mediterranean countries. The most affected organ is the liver, however hydatidosis can be found anywhere in the human body.MethodsThe records of patients who were diagnosed with hydatid disease in our hospital from December 2005 to February 2016 were analyzed retrospectively. The cases were evaluated and recorded depending on their gender, age and the localization of the cysts.ResultsA total of 329 patients diagnosed over a 10-year period were included in our study. There were 202 females (61.4%) and 127 males (38.6%). The hydatid cysts were located in the liver in 257 (78.1%) patients and in unusual locations in 72 (21.9%) patients. The most common unusual site for hydatid cysts was the spleen followed by bones, central nervous system, soft tissue, the kidney and the gall bladder. Amongst these 72 patients who had hydatid cysts in unusual locations; 33 patients had concomitant liver hydatidosis, whereas 39 patients had primary involvement of unusual sites. Two patients with malignancies also had hydatid cysts in different locations.ConclusionHydatid disease affects many organs in the body and therefore it can pose a major diagnostic dilemma and it may mimic other entities. In endemic areas, a differential diagnosis of hydatid disease should be considered for cystic masses in any anatomical location.     

Phylogenetic analysis of rubella viruses isolated in 2008 outbreak in Argentina

BackgroundA rubella outbreak was recorded in Buenos Aires during 2008.ObjectivesThe objective of this communication is to present the genetic and phylogenetic analyses of wild-type RUBV circulating in Buenos Aires during the 2008 outbreak.Study designThroat swab samples collected from patients diagnosed with rubella between June 2008 and December 2008 were inoculated in cell culture and 23 isolates were sequenced.ResultsPhylogenetic analysis of the WHO-recommended window (nt 8731–9469) of the E1 envelope glycoprotein was performed and all isolates clustered with the 2B genotype.ConclusionsGenotype 2B seems to be endemically circulating in the Southern cone of Latin America, thus causing recent outbreaks.     

In vitro and in vivo treatments of Echinococcus granulosus with Huaier aqueous extract and albendazole liposome

The aim of this study was to investigate the in vitro and in vivo efficacies of chemotherapy employing albendazole liposome (L-ABZ), Huaier aqueous extract, and a Huaier aqueous extract/L-ABZ combination against Echinococcus granulosus. Protoscolices of E. granulosus were incubated in vitro with the two drugs, either separately or in combination, at the following final concentrations: 2 mg/mL Huaier aqueous extract, 10 μg/mL L-ABZ, and 2 mg/mL Huaier aqueous extract + 10 μg/mL?L-ABZ. Huaier aqueous extract and L-ABZ displayed slower protoscolicidal activity when applied separately than when used in combination. The maximum protoscolicidal effect was found with the combination Huaier aqueous extract + L-ABZ. Despite the low Huaier aqueous extract + L-ABZ concentrations used, protoscolex viability dropped rapidly. In vivo studies were performed on mice injected with protoscolices of E. granulosus. Huaier aqueous extract and L-ABZ were administered three times a week for a period of 4 months by the oral route. Huaier aqueous extract in E. granulosus-infected mice was effective. Combined application of both drugs did increase the treatment efficacy. In conclusion, the outcomes obtained clearly demonstrated that in vitro and in vivo treatment with Huaier aqueous extract and L-ABZ is effective against E. granulosus.     

Prevalence of hepatitis C virus infection according to the year of birth: identification of risk groups

Hepatitis C virus (HCV) screening according to the year of birth is recommended is some countries based on epidemiological data. The aim of this study was to analyze anti-HCV prevalence among people born between 1905 and 2015 in Argentina. Patients attending a tertiary care hospital in Buenos Aires, Argentina, from 2001 to 2015, who had a determination of anti-HCV, were included. Of 22,079 patients analyzed, 1,152 (5.2%; 95% confidence interval [CI]: 4.9%–5.5%) patients showed positive anti-HCV and 729 (3.3%; 95% CI: 3.1%–3.5%) patients showed detectable viremia. Three risk groups were identified (HCV prevalence): low-risk group—outpatient clinics/emergencies (2.8%); intermediate-risk group—in-patients (8%); and high-risk group—dialysis/transplants (27.2%). In the low-risk group, being born in 1973 or before was identified as a cut-off value for the risk of anti-HCV acquisition (area under the receiver-operator characteristic curve: 75.1 [95% asymptotic CI: 0.732–0.770; p < 0.001]). Ninety-one patients born after 1973 (0.8%) showed positive anti-HCV versus 457 individuals born in 1973 or before (5.8%), p < 0.001. In this group, positive anti-HCV was observed in 252 females (2.1%) and 296 males (4.1%), p < 0.001. In a multivariate analysis adjusted for gender, alanine-aminotransferase levels and HIV coinfection, being born in 1973 or before was independently identified as a risk for positive anti-HCV (adjusted odds ratio: 14.234 [95% CI: 9.993–20.277]; p < 0.001). People born in 1973 or before without other risk factors should be included in screening programs to link the highest possible number of HCV-infected patients to appropriate care and treatment.     

人体包虫病主要特异性免疫球蛋白的水平及其临床意义

本文报道了应用酶联免疫吸附试验(ELISA)检测包虫病人血清中抗包虫特异性免疫球蛋白(IgG和IgM)的水平并对其临床意义进行了初步探讨。48例正常人血清特异性IgG和IgM的假阳性检出串均为0％;24例囊尾蚴病人血清特异性抗包虫IgG和IgM的假阳性检出效分别为:33.4％;4.16％;42例包虫病患者血清特异性IgG和IgM的阳性检出率分别为:80.95％,66.67％;两者之差经统计学处理具有显著意义(p<0.05)。肝包虫特异性IgG的阳性检出率较肺包虫高,肺包虫特异性IgM的阳性检出率较肝包虫高。这提示患者对包虫的免疫应答和其寄生的位置有关;诱导产生的特异性免疫球蛋白主要为IgG类,特异性IgM的水平与包虫囊壁的完整与否以及检测方法的灵敏性、特异性有关。     

Application of recombinant Echinococcus granulosus antigen B to ELISA kits for diagnosing hydatidosis

Echinococcus granulosus causes human cystic echinococcosis as an important public health problem in many regions of the world. There are some problems in primary diagnosis such as cross-reaction with sera from patients with other parasitic disease in serological tests. The use of an appropriate source of antigenic material is a very important and crucial point in the improvement of the serodiagnostic features such as enzyme-linked immunosorbent assay (ELISA) method. We expressed and purified recombinant AgB of Echinococcus granulosus and used as antigen in ELISA method. Serum samples were given from 36 cystic hydatid disease patients that have been confirmed by surgical operation as well as 36 healthy individuals sera were tested by ELISA method using recombinant AgB and compared with commercial kit (Euroimmun) for specificity and sensitivities value. The sensitivity of 91.66% and specificity of 97.22% were determined by homemade kit.