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Cystic echinococcosis (CE) is an emergent neglected disease affecting human and animals in Egypt with a wide distribution and incidence. This study aimed to evaluate the use of a polyclonal antibody-based sandwich ELISA in the detection of Echinococcus granulosus antigen in human and camel sera. Hydatid cyst protoscoleces antigen (PsAg) was isolated from hydatid cysts collected from naturally infected camel livers and lungs. PsAg was used for immunization of rabbits to raise IgG polyclonal antibodies (IgG PsAb). IgG PsAb were then precipitated, purified using Protein-A Sepharose gel and labeled with horseradish peroxidase enzyme. We assayed the purity of the IgG PsAb, and the two prepared E. granulosus antigens CPsAg from camel cysts and HPsAg from human cysts by Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The resulted protein bands of the prepared CPsAg appeared at different molecular weights: 180, 90, 68, 54, 42 and 22 kDa while, HPsAg shared with it in 4 common bands at 68, 54, 42, and 22 kDa. The purified IgG PsAb had been resolved at two bands at 52 kDa and at 32 kDa. Sandwich ELISA were performed for the detection of circulating E. granulosus antigens in sera of human (n = 183) and camels (n = 190). The purified IgG PsAb showed strong reactivity against E. granulosus infected human and camel samples and no cross reactivity neither with free-healthy negative sera nor with others parasitic diseases (Schistosomiasis, Fascioliasis, Toxoplasmosis, Ancylostomiasis for human samples and Fascioliasis, ticks' infestation, Eimeriosis, Cryptosporidiosis, Nasal myiasis, Toxoplasmosis for camel samples). The sensitivity of the assay was 98.25% (56/57) and 96.9% (31/32) against human and camel samples, respectively. Specificity was 100% in both human and camel samples. Sandwich ELISA detected CE in 33.3% (24/72) and 55.6% (50/90) random human and camel samples, respectively. Indirect ELISA, using CPsAg, was used for detection of antibodies in positive human and camels' sera and detected 96.5% (55/57) and 93.8% (30/32) of human and camel samples, respectively. In our study, Genomic DNA was extracted from protoscoleces fluid of human liver hydatid cysts to identify the Echinococcus sp. isolate based on NADH dehydrogenase subunit 1 (NAD1) gene by Polymerase Chain Reaction (PCR) and the isolate (GenBank: OP785689.1) were identified as E. granulosus sensu lato genotype. In conclusion, Sandwich ELISA technique was found to be a potent and sensitive assay for detection of hydatid antigen in both human and camel samples., (© 2024. The Author(s).)

Purpose: Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus. Clinical, radiologic, pathologic, and serologic findings should be evaluated together for the diagnosis of CE. The sensitivity and specificity oalf serologic tests may vary depending on the method used. In this study, we aimed to detect IgG antibodies specific to E. granulosus using indirect hemagglutination assay (IHA), enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibodies (IFA) and western blot (WB) tests., Methods: In our study, the serum samples of 74 patients sent to our laboratory with suspicion of CE were studied using two different commercial IHA tests, ELISA, IFA and WB test. The test results were evaluated along with radiological findings and histopathological examinations, the latter being the gold standard., Results: Of all the patients, 51 (69%) were female and 23 (31%) were male. There was a statistically significant difference between males and females (χ 2  = 9.7, p = 0.002). Out of 74 patients, positivity rates for Siemens IHA, Fumouze IHA, ELISA, IFA and WB test were positive as 33 (44.6%), 35 (47.3%), 43 (58.1%), 42 (56.7%) and 38 (51.3%), respectively. The sensitivity and specificity of the tests were as follows: 66.67 and 2.31% for Siemens IHA; 70.83% and 96.15% for Fumouze IHA; 85.42%, and 88.46% for ELISA; 83.33% and 88.46% for IFA; 72.92% and 88.46% for WB test., Conclusion: There were statistically significant differences in between all five methods (p < 0,001). While the tests with the highest specificity was Fumouze IHA, the test with the highest sensitivity was the ELISA test. It was concluded that IHA and ELISA tests were more practical in practice because of their greater applicability., (© 2024. The Author(s).)

Both E. multilocularis and host-derived exosomes are involved in the pathogenic process of alveolar echinococcosis (AE). Exosomes secrete miRNAs that have regulatory roles in host-pathogen interactions in multiple ways. In the present study, we collected and purified supernatants of E. multilocularis cultures, as well as human plasma exosomes. High-throughput sequencing showed the identities of 45 exosomal miRNAs in E. multilocularis. The lengths of these miRNAs ranged from 19 to 25 nucleotides (nt), with the majority (n = 18) measuring 22 nt. Notably, emu-let-7-5p emerged as the most abundant among these miRNAs, with a detected count of 33,097 and also length of 22 nt. Nanoparticle tracking analysis (NTA) showed that the concentration of exosomes in the plasma of AE patients was lower compared to that in the healthy individuals. This result suggested that the concentration of plasma exosomes was able to distinguish AE patients from healthy individuals. Using qRT-PCR to assess the relative expression of 10 miRNAs of E. multilocularis, we showed that the expression of miR-184-3p was downregulated significantly in the exosomes of plasma from AE patients compared to that in the control group. In summary, this study indicates that AE induces a reduction in the concentration of human plasma exosomes, as well as downregulating miR-184-3p in infected individuals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Introduction: Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the metacestode of Echinococcus granulosus. CE is a health problem in Middle Eastern countries, such as Iran. The purpose of this study was to purify subunit 8 KDa antigen B from crude sheep hydatid cyst fluid (HCF) and compare its sensitivity and specificity with a commercial human ELISA kit (PT-Hydatid-96)., Methods: 28 sera samples were collected from hydatid cyst patients who had surgery for a hydatid cyst and had their disease confirmed by pathology after the surgery. Furthermore, 35 samples of healthy individuals with no history of hydatid cysts were collected, as were nine serum samples from parasite-infected non-CE patients. HCF was obtained from sheep fertile cysts at a Sari slaughterhouse and used as an antigen. In an indirect ELISA test, the B antigen was employed, and the results were compared to those from a commercial ELISA kit., Results: The results of this study were analyzed using the Kappa test. The commercial ELISA kit showed 17 cases (23.6%) positive, 44 cases (61.1%) negative, and 11 cases (15.3%) borderline. B antigen showed that 18 (25%), 43 (59.7 %), and 11 (15.3%) were positive, negative, and borderline, respectively. One sample (1.4% of 72 total samples) of 35 serum samples from healthy individuals was positive using B antigen-based ELISA. In addition, all nine serum samples from parasite-infected non-CE patients were negative for both tests. The sensitivity and specificity of the commercial ELISA kit have been evaluated at 60.7% and 100%, respectively. For B antigenbased ELISA, these values are 64.3 and 97.7%, respectively., Conclusion: Antigen B produced from hydatid cyst fluid is a promising option for serological identification of hydatid cysts in both infected and healthy individuals. In an indirect ELISA test, hydatid fluid antigen could be used as a precise source of detection., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Cystic Echinococcosis (CE) is the silent chronic Helminthes zoonotic infection caused by the larval stage in intermediate hosts of the dog tapeworm Echicoccous granulosus , which belongs to the Taeniidae family and genus Echinococcu ssp . According to the study of CE, the hospitalization and surgeries of patients indicated the high prevalence of the disease in Iraq.This study aimed to determine some immunological parameters in patients infected with Echinococcus granulose . The study of 53 patients infected with CE showed that in 40 (75.4 %), 5 (9.4%), 4 (7.5%), and 2 (3.7 %) cases the liver, abdominal cavity, kidney, and lungs were involved. In terms of age, most and least number of the patients were within the age ranges of30-40 (n=15, 28.3 %) and60-70 years old (n=7, 13.2 %), respectively. Moreover, 37(56.9%) and 16(43%) of them were female and male, respectively. The enzyme-linked immunosorbent assay (ELISA) was used to measure the level of the Interleukin family among patients. There was a significant increase in the serum level concentration of IL17A and IL17B in patients with hydatid disease, compared to the control group. The changes in different age groups also showed statistically significant differences among them ( P ≤0.05). The outcome of this study indicated that CE is endemic in Babylon province, Iraq. The ELISA technique is a reliable and efficient test for the early diagnosis and monitoring of human hydatid disease. Moreover, it was found that the liver was the most common site of human hydatid cyst.

Objective: CD8 + T cells can participate in immune action by secreting various cytokines, which have a killing effect on certain viruses, tumor cells, and other antigenic substances. However, in studies such as chronic viral infections and some parasitic infections, CD8 + T lymphocyte showed functional depletion, and its immune dysfunction was an important reason for the persistence of infection. Tim-3 has been shown to be a negative regulator of CD8 + T cell function, causing depletion of CD8 + T cells in cancer and chronic infection. However, the relationship between Tim-3 and CD8 + T cells in Echinococcus multilocularis infection is not clear., Methods: In this study, we analyzed peripheral blood CD8 + T cells from 62 alveolar echinococcosis (AE) patients and 30 healthy controls., Results: Compared with the healthy control group, the proportion of CD8 + T cells in the peripheral blood of AE patients increased significantly, while the levels of perforin, granzyme B and IFN-γ in peripheral blood CD8 + T cell related factors of metabolically active alveolar echinococcosis (MAAE) patients decreased significantly. Later detection revealed that the expression of Tim-3 on CD8 + T cells in the peripheral blood of MAAE patients was significantly higher than that of metabolically inactive alveolar echinococcosis (MIAE) patients and healthy controls. The expression levels of function-related factors perforin, granzyme B and IFN-γ in CD8 + Tim-3 + T cell were significantly lower in the CD8 + Tim-3 - T cells of AE patients. In vitro, the secretion of CD8 + T cell-associated factors was significantly restored by inhibiting Tim-3 expression., Conclusion: Therefore, the depletion of CD8 + T lymphocyte in patients with alveolar echinococcosis disease is considered to be related to the high expression of Tim-3 on the surface., (© 2021 by the Association of Clinical Scientists, Inc.)

Background: Cystic echinococcosis (CE) is a serious parasitic zoonosis caused by the larvae of the tapeworm Echinococcus granulosus. The development of an effective vaccine is one of the most promising strategies for controlling CE., Methods: The E. granulosus 3-hydroxyacyl-CoA dehydrogenase (EgHCDH) gene was cloned and expressed in Escherichia coli. The distribution of EgHCDH in protoscoleces (PSCs) and adult worms was analyzed using immunofluorescence. The transcript levels of EgHCDH in PSCs and adult worms were analyzed using quantitative real-time reverse transcription PCR (RT-qPCR). The immune protective effects of the rEgHCDH were evaluated., Results: The 924-bp open reading frame sequence of EgHCDH, which encodes a protein of approximately 34 kDa, was obtained. RT-qPCR analysis revealed that EgHCDH was expressed in both the PSCs and adult worms of E. granulosus. Immunofluorescence analysis showed that EgHCDH was mainly localized in the tegument of PSCs and adult worms. Western blot analysis showed that the recombinant protein was recognized by E. granulosus-infected dog sera. Animal challenge experiments demonstrated that dogs immunized with recombinant (r)EgHCDH had significantly higher serum IgG, interferon gamma and interleukin-4 concentrations than the phosphate-buffered saline (PBS) control group. The rEgHCDH vaccine was able to significantly reduce the number of E. granulosus and inhibit the segmental development of E. granulosus compared to the PBS control group., Conclusions: The results suggest that rEgHCDH can induce partial immune protection against infection with E. granulosus and could be an effective candidate for the development of new vaccines., (© 2021. The Author(s).)

Stereocontrolled syntheses of biotin-labeled oligosaccharide portions containing the carbohydrate moiety of glycoprotein from Echinococcus granulosus have been accomplished. Trisaccharide Galβ1-3Galβ1-3GalNAcα1-R ( A ), tetrasaccharide Galα1-4Galβ1-3Galβ1-3GalNAcα1-R ( B ), and pentasaccharide Galα1-4Galβ1-3Galβ1-3Galβ1-3GalNAcα1-R ( C ), (R = biotinylated probe) were synthesized by stepwise condensation and/or block synthesis by the use of 5-(methoxycarbonyl)pentyl 2-azido-4,6- O -benzylidene-2-deoxy-α-d-galactopyranoside as a common glycosyl acceptor. The synthesis of the tetrasaccharide and the pentasaccharide was improved from the viewpoint of reducing the number of synthetic steps and increasing the total yield by changing from stepwise condensation to block synthesis. Moreover, hexasaccharide E , which contains the oligosaccharide sequence which occurs in E. granulosus , was synthesized from trisaccharide D . We examined the antigenicity of these five oligosaccharides by an enzyme-linked immunosorbent assay (ELISA). Although compounds of C - E did not exhibit antigenicity against cystic echinococcosis (CE) patient sera, compounds B , D , and E showed good serodiagnostic potential for alveolar echinococcosis (AE).

Background: Cystic echinococcosis (CE) is a complex disease for which clear understanding of clinical manifestations is needed to avoid misdiagnosis, inappropriate treatment, and severe complications. We evaluated the accuracy of a whole-blood stimulation test based on Interleukin (IL)-4 detection in response to Antigen B (AgB) of Echinococcus granulosus sensu lato to discriminate cyst viability and detect cyst reactivation in patients with hepatic CE., Methodology/principal Findings: Thirty patients with CE3b cysts and 37 patients with spontaneously-inactivated CE4-CE5 cysts were recruited (T0). After enrollment, 5 patients with CE3b cysts received albendazole, resulting in cyst solidification (CE4) in 4/5. Within a two-year follow-up, the whole-blood test was repeated at two time-points, in ≥14 (T1) and in ≥4 (T2) patients per group. IL-4 and a panel of other soluble factors were measured in the stimulated plasma. Baseline IL-4 levels were significantly higher in patients with CE3b compared to those with CE4 cysts (p = 0.006). Test accuracy for CE3b diagnosis had a sensitivity of 33-60% and a specificity of 76-95%, depending on the cut-off applied. Overall, IL-4 levels did not change significantly over time in either group; however, patients within the CE3b group showed a significant decrease of IL-1ra, IL-6, IL-8, G-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, FGF at T1 compared to T0 (p≤0.042)., Conclusions/significance: Whole-blood IL-4-response to AgB is significantly higher in patients with active compared to inactive CE but apparently not modulated over time after treatment. On the contrary, the levels of IL-1ra, IL-6, IL-8, G-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, FGF significantly decreased in active CE during follow-up. Additional studies are needed to understand whether these findings might have a clinical significance for patients' follow-up., Competing Interests: The authors have declared that no competing interests exist.

All the time, echinococcosis is a global zoonotic disease which seriously endangers public health all over the world. In order to speed up the development process of anti- Echinococcus granulosus vaccine, at the same time, it can also save economic cost. In this study, immunoinformatics tools and molecular docking methods were used to predict and screen the antigen epitopes of Echinococcus granulosus , to design a multi-epitope vaccine containing B- and T-cell epitopes. The multi-epitope vaccine could activate B lymphocytes to produce specific antibodies theoretically, which could protect the human body against Echinococcus granulosus infection. It also could activate T lymphocytes and clear the infected parasites in the body. In this study, four CD8 + T-cell epitopes, three CD4 + T-cell epitopes and four B-cell epitopes of Protein EgTeg were identified by immunoinformatics methods. Meanwhile, three CD8 + T-cell epitopes, two CD4 + T-cell epitopes and four B-cell epitopes of Protein EgFABP1 were identified. We constructed the multi-epitope vaccine using linker proteins. The study based on the traditional methods of antigen epitope prediction, further optimized the prediction results combined with molecular docking technology and improved the precision and accuracy of the results. Finally, in vivo and in vitro experiments had verified that the vaccine designed in this study had good antigenicity and immunogenicity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Yu, Zhu, Li, Chen, Sha, Li, Zhang and Ding.)

Introduction: Cystic echinococcosis, caused by Echinococcus granulosus, is a neglected zoonosis that affects humans and livestock. This sero-survey was designed for the first time in Pakistan to assess the exposure of butchers to E. granulosus as there was no previous report in the country for this occupational group., Methodology: Blood samples were collected from registered butchers (n = 364) in five different slaughterhouses in Faisalabad and Bahawalnagar Districts. Sera were tested for anti-Echinococcus granulosus IgG with a commercially available ELISA kit (specificity, 100%; sensitivity, 97%)., Results: Overall, seroprevalence was 9.61% (35/364). Butchers >30 years of age (10.34%), those involved in small ruminants butchery (11.70%), >10 years' experience (10.04%), formal education level up to middle standard (10.28%), contact with dogs (12.71%), improper/unhygienic disposal of dog feces (11.87%), and those unaware of the consequences of eating with unwashed hands (13.80%) were more seropositive with significant statistical differences (p < 0.05). Variables like previous cyst encounter, no knowledge of zoonoses and/or cystic echinococcosis, living in rural areas and the presence of stray/feral dogs in surroundings did not show any significant association (p > 0.05) with seroprevalence in butchers. The binary logistic regression model also showed a statistically significant relationship (p < 0.05) for all risk factors found statistically significant (p < 0.05) in the univariate analysis., Conclusions: This study shows high prevalence of cystic echinococcosis among butchers in Pakistan and underscores the need for educating native slaughterhouse personnel on cystic echinococcosis. It also serves as a global warning, especially in developing countries., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2021 Mughees Aizaz Alvi, Li Li, Muhammad Saqib, John Asekhaen Ohiolei, Muhammad Waqar Younas, Muhammad Haleem Tayyab, Ali Hassan, Warda Qamar, Anum Aizaz Alvi, Bao-Quan Fu, Hong-Bin Yan, Wan-Zhong Jia.)

Background: Cystic echinococcosis (CE) affects predominantly young patients in highly endemic areas. Improved serological methods are needed for the follow-up of CE cases, especially given the high rates of post-surgical relapse that require detection as soon as possible., Methods: We designed a study to investigate the value of antigenic proteins extracted from Echinococcus granulosus (E. granulosus) protoscoleces, and of recombinant B2t and 2B2t proteins, for assessing the efficacy of surgical treatment carried out on CE-affected children. This study was performed on 278 plasma samples collected from 59 Tunisian children surgically treated for CE and monitored for 3 years post-surgery. The patients were classified according to post-surgical outcomes into a "non-relapsed" (NRCE) and a "relapsed" (RCE) group. We performed in-house ELISAs to measure anti-B2t and anti-2B2t IgG and immunoblotting for the detection of IgG against SDS-PAGE-resolved E. granulosus protoscoleces-specific antigens. The Wilcoxon test was applied to assess anti-B2t and anti-2B2t IgG levels. We applied the Cochran Q test to compare the distribution of immunoblotting antigenic bands between 1-month and 1-year post-surgery., Results: The probability of being "relapse-free" when a decrease in antibody titers occurred between 1 month and 1 year post-surgery was 81% and 75%, respectively, for anti-B2t and anti-2B2t IgG. We identified five protoscolex protein bands of 20, 26/27, 30, 40 and 46 kDa as highly immunoreactive by immunoblot for both RCE and NRCE patients at 1 month post-surgery, and significantly lower immunoreactivity after 1 year (p < 10 -4 ) for NRCE compared to RCE patients. The proteins at 26/27 and 40 kDa displayed the best performance in predicting the outcome, with an 84% probability of being relapse-free when the reactivity against the 40 kDa antigen, the doublet at 26/27 kDa, or both was absent or disappeared between 1 month and 1 year post-surgery, and a 93% probability of being relapsed when both bands remained reactive or increased in intensity between the two time points., Conclusions: The B2t protein could be useful for the prediction of CE early post-surgical outcomes. The proteins of E. granulosus protoscoleces, especially the doublet P26/27 and P40, could be promising predictive biomarkers for the post-surgical follow-up of CE cases as well.

Background: Alveolar echinococcosis (AE) is caused by parasitic infection by Echinococcus multilocularis. Its diagnosis is usually based on clinical symptoms, ultrasound, and other imaging methods. MicroRNAs (miRNAs) play important roles in disease processes and can exist in a highly stable cell-free form in body fluids. It is important to identify specific, sensitive diagnostic markers for early diagnosis and evaluation of AE. In this study, we examined hsa-miR-125b-5p as a potential plasma biomarker of E. multilocularis infection., Methods: Plasma samples from patients with AE and healthy individuals were screened for the presence of five miRNAs using miRNA chips. We used quantitative polymerase chain reaction to measure miRNA expression levels in plasma and liver tissue samples from patients with AE., Results: hsa-miR-125b-5p was stably upregulated in the plasma and liver tissue samples from patients with AE., Conclusions: The results suggest that hsa-miR-125b-5p may be a promising biomarker for early, non-invasive diagnosis of AE.

Over one million people are living with cystic echinococcosis (CE) and alveolar echinococcosis (AE). For CE, long-term albendazole treatment is often needed, which requires regular follow-up. Follow-up is mainly through imaging which is insensitive to subtle changes and subjective to experience. We investigated the changes of Echinococcus granulosus (Eg) cell-free DNA (cfDNA) in plasma of CE patients before and after albendazole treatment to evaluate its potential as an objective marker for treatment follow-up. Plasma samples of nine CE patients were collected before and after treatment. We identified Eg cfDNA from every sample through high-throughput sequencing. Eg cfDNA concentration and fragment length increased significantly after the treatment period. Ultrasound examination before and after the treatment initiation reflected the drug effects to a certain extent, as the cyst size of four patients reduced. Our findings indicated that Eg cfDNA from plasma could be a potential marker in the monitoring of CE treatment., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Objective: Cystic echinococcosis (CE) is a zoonotic disease, which leads to morbidity and mortality worldwide. This study aimed to retrospectively evaluate the presence of anti- Echinococcus granulosus immunoglobulin G (IgG) antibodies, which were detected by indirect fluorescent antibody test in the samples that were transferred to the Microbiology Laboratory of University of Health Sciences Turkey, Diyarbakır Gazi Yaşargil Training and Research Hospital with the pre-diagnosis of CE. Moreover, gender differences with respect to positivity rates of anti- E. granulosus IgG antibodies were investigated., Methods: Anti- E. granulosus IgG antibodies, which were detected in the samples of cases with the pre-diagnosis of CE between January 2014 and December 2017, were retrospectively evaluated. Gender difference with respect to positivity rates was investigated by applying the chi-square test in cases with positive anti- E. granulosus IgG antibodies., Results: Out of the 829 serum samples, 222 (26.7%) were found to be positive for E. granulosus IgG antibodies, among which 40 (27.2%), 56 (25.5%), 51 (23.3%) and 75 (30.6%) were found to be positive in 2014, 2015, 2016 and 2017, respectively. No significant difference was found between genders amongst the cases with positive anti- E. granulosus IgG antibodies (p>0.05)., Conclusion: Since CE is a major public health problem, evaluation of the presence of anti- E. granulosus IgG antibodies would be important to understand the positivity rate at the regional level.

Objective: Describe the case of echinococcal disease in gynecological practice and point out the complications of its diagnosis., Design: Case report., Setting: Department of Obstetrics and Gynaecology, Faculty Hospital Tren&#269;ín, Slovakia., Case Report: The case is presented in an immunosuppressed patient with peritoneal carcinomatosis, mild Ca 125 elevation and increased CRP presenting as a finding of gynecological etiology with histopathological outcome and conclusion of parasitic disease - echinococcosis (hydatidosis)., Conclusion: In the differential diagnosis of peritoneal carcinomatosis and ascites, especially in immunosuppressed patients with a positive social (or epidemiological) history, the possibility of rare parasitic diseases such as echinococcosis, which resembles malignant tumors, should be considered.

Background: Cystic echinococcosis (CE) is a widespread parasitic disease caused by the larval stage of Echinococcus granulosus . Since current methods for the diagnosis of CE are not efficient enough, rapid, and reliable tests are required for the acceleration of CE diagnosis. The present study aimed to produce recombinant B8/1 and B8/2 antigens of E. granulosus and evaluate their sensitivities and specificities separately and simultaneously for the diagnosis of CE., Methods: The recombinant B8/1 and B8/2 antigens were produced and used in an ELISA system for the diagnosis of CE. The sera specimens including 30 sera from pathologically confirmed CE patients, 30 from other non-CE patients, and 30 from healthy controls, were evaluated by the ELISA, using AgB8/1 and AgB8/2., Results: The results showed a sensitivity of 93.33%, 90%, and 96.7% for AgB8/1, AgB8/2, and their combination, respectively. The specificities were 91.7%, 93.33%, and 93.33% for AgB8/1, AgB8/2, and their combination, respectively., Conclusion: Simultaneous usage of AgB8/1 and AgB8/2 increased the test sensitivity for the diagnosis of CE. Furthermore, the specificity of AgB8/1 and AgB8/2 combination was more than AgB8/1 and equal to AgB8/2 alone. The findings revealed that the simultaneous usage of AgB8/1 and AgB8/2 could be a suitable approach for the diagnosis of CE.

The reference diagnostic method of human abdominal Cystic Echinococcosis (CE) is imaging, particularly ultrasound, supported by serology when imaging is inconclusive. However, current diagnostic tools are neither optimal nor widely available. The availability of a test detecting circulating biomarkers would considerably improve CE diagnosis and cyst staging (active vs inactive), as well as treatments and follow-up of patients. Exosomes are extracellular vesicles involved in intercellular communication, including immune system responses, and are a recognized source of biomarkers. With the aim of identifying potential biomarkers, plasma pools from patients infected by active or inactive CE, as well as from control subjects, were processed to isolate exosomes for proteomic label-free quantitative analysis. Results were statistically processed and subjected to bioinformatics analysis to define distinct features associated with parasite viability. First, a few parasite proteins were identified that were specifically associated with either active or inactive CE, which represent potential biomarkers to be validated in further studies. Second, numerous identified proteins of human origin were common to active and inactive CE, confirming an overlap of several immune response pathways. However, a subset of human proteins specific to either active or inactive CE, and central in the respective protein-protein interaction networks, were identified. These include the Src family kinases Src and Lyn, and the immune-suppressive cytokine TGF-β in active CE, and Cdc42 in inactive CE. The Src and Lyn Kinases were confirmed as potential markers of active CE in totally independent plasma pools. In addition, insights were obtained on immune response profiles: largely consistent with previous evidence, our observations hint to a Th1/Th2/regulatory immune environment in patients with active CE and a Th1/inflammatory environment with a component of the wound healing response in the presence of inactive CE. Of note, our results were obtained for the first time from the analysis of samples obtained in vivo from a well-characterized, large cohort of human subjects., Competing Interests: The authors have declared that no competing interests exist.

E. granulosus is a cestode that causes Cystic Echinococcosis (CE), a zoonotic disease with worldwide presence. The immune response generated by the host against the metacestode induces a permissive Th2 response, as opposed to pro-inflammatory Th1 response. In this view, mixed Th2 and regulatory responses allow parasite survival. Overall, larval Echinococcus infections induce strong regulatory responses. Fasciola hepatica, another common helminth parasite, represents a major infection in cattle. Co-infection with different parasite species in the same host, polyparasitism, is a common occurrence involving E. granulosus and F. hepatica in cattle. 'While it is known that infection with F. hepatica also triggers a polarized Th2/Treg immune response, little is reported regarding effects on the systemic immune response of this example of polyparasitism. F. hepatica also triggers immune responses polarized to the Th2/ Treg spectrum. Serum samples from 107 animals were analyzed, and were divided according to their infection status and Echinococcal cysts fertility. Cytokines were measured utilizing a Milliplex Magnetic Bead Panel to detect IFN-γ, IL-1, IL-2, IL-4, IL-6, IL-10, IL-12 and IL-18. Cattle infected only with F. hepatica had the highest concentration of every cytokine analyzed, with both 4.24 and 3.34-fold increases in IL-10 and IL-4, respectively, compared to control animals, followed by E. granulosus and F. hepatica co-infected animals with two-fold increase in IL-10 and IL-4, compared to control animals, suggesting that E. granulosus co-infection dampens the cattle Th2/Treg immune response against F. hepatica. When considering Echinococcal cyst fertility and systemic cytokine concentrations, fertile cysts had higher IFN-γ, IL-6 and IL-18 concentrations, while infertile cysts had higher IL-10 concentrations. These results show that E. granulosus co-infection lowers Th1 and Th2 cytokine serological concentration when compared to F. hepatica infection alone. E. granulosus infections show no difference in IFN-γ, IL-1, IL-2, IL-6 and IL-18 levels compared with control animals, highlighting the immune evasion mechanisms of this cestode., Competing Interests: The authors have declared that no competing interests exist.

Objective: To determine the results of radiological and serological screenings in individuals who shared the same living space as patients with hydatid cyst in a State Hospital of Afghanistan., Methods: Patients presenting with hydatid cyst to a public hospital in Afghanistan were included in this study. Full sampling method was used. Also, the relatives of the patients were called to the hospital and investigated for the presence of hydatid cyst antibodies through direct chest X-ray, upper abdominal ultrasonography and ELISA., Results: During the study period, a total of 214 patients, including 102 male and 112 female, underwent radiological and serological screenings. While cysts were radiologically detected in the liver, lung and spleen in 8, 2 and 1 patient, respectively, the serology was positive in 22 patients., Conclusion: As a result of the study, it was concluded that the patients who shared the same environment as the patients who were diagnosed with hydatid cyst in an endemic region for cyst hydatid disease.

The development of suitable serological tests for the diagnosis of CE is still necessary. This study aimed to evaluate the efficacy of ELISA in the diagnosis of human cystic echinococcosis (CE), using parasite protoscolices antigens. Liver hydatid cysts were isolated from sheep infected with hydatid cysts and the protoscolices were isolated from the hydatid cyst fluid. Protoscolices crude antigen was prepared by mechanical disruption, plus freeze-thawing and sonication methods. Thirty sera samples of confirmed hydatid cyst patients, 30 samples of healthy individuals, and 30 samples of people with other infections were collected and the samples were evaluated in an ELISA system, using the crude protoscolices antigen. The sera samples were also simultaneously evaluated by antigen B-ELISA. The estimated value of sensitivity and specificity for the ELISA, using the crude protoscolices antigens, was 93.3% (95% CI: 76.4-98.8%) and 90% (95% CI: 78.8-95.8%), respectively. These values were 86.6 (95% CI: 68.3-95.6) and 91 (95% CI: 80.81-96.9) for the antigen-B based ELISA. Antigens prepared from protoscolices of hydatid cyst are suitable candidates for the serologic diagnosis of human CE. Further studies are needed to identify a single specific antigen among the protoscolices antigens to improve the diagnostic performance of these antigens.

Background: Alveolar echinococcosis (AE) is a zoonotic parasitic disease caused by Echinococcus multilocularis larval tapeworm infections in humans that severely impairs the health of affected patients in the northern hemisphere., Methods: The expression levels of 20 cytokines associated with AE infection were measured by enzyme-linked immunosorbent assay, and the correlations between these cytokines were analysed in the R programming language., Results: Serum cytokine levels differed among individuals in both the AE patient and healthy control groups. The results of the correlations among the cytokines showed obvious differences between the two groups. In the AE patients group, Th1 and Th2 cytokines formed a more complicated network than that in the healthy control group., Conclusions: The altered correlations between Th1 and Th2 cytokines may be closely associated with AE infection, which may provide a new explanation for the essential differences between AE patients and healthy individuals.

Currently, cystic echinococcosis (CE) follow-up is a serious concern among surgeons. MicroRNAs (miRNAs) are small, endogenous, non-coding RNAs which are present in human body fluids in a highly stable form. Recently, it is observed that Echinococcus granulosus expresses a large number of miRNAs in its developmental stages. The current study aimed at evaluating the capacity of parasitic miRNAs to serve as plasma biomarkers for hydatid cysts before and after CE surgery. Hydatidosis patients were identified using radiological and histopathological examinations. Following RNA extraction and cDNA synthesis, the expression levels of parasite-derived miRNAs including egr-miR-71 and egr-let-7 were quantitatively evaluated using real-time polymerase chain reaction (RT-PCR) in 30 hydatid cyst-infected individuals before surgery and an equal number of healthy controls. Then, three- and six-month follow-ups were performed after cystectomy. To analyze parasite-derived miRNAs, the relative fold change between uninfected and infected samples was determined and normalized to hsa-miR-16-5p as the housekeeping internal control. RT-PCR demonstrated that egr-miR-71 and egr-let-7 were specifically amplified in all the plasma samples from the infected individuals with hydatid cyst; yet they were significantly down-regulated at three and six months' post-surgery (P < 0.05). The egr-miR-71 had a higher level of expression in larval stage compared with egr-let-7. The results of the current study indicated that hydatid cyst-derived miRNAs including egr-miR-71 and egr-let-7 can be detected in human plasma. Considering the changes in the expression levels of these miRNAs after three and six months, it seems that these miRNAs, especially egr-miR-71, could serve as novel promising biomarkers for the early diagnosis and monitoring of hydatidosis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Echinococcosis is a serious zoonotic parasitic disease that could be fatal without diagnosis and treatment in a timely manner. Herein, we present a rapid and label-free method for screening of echinococcosis using surface-enhanced Raman spectroscopy (SERS). Three groups of serum SERS spectra based on porous silicon/silver composites are obtained: one group from healthy volunteers (normal, n = 163) and two other groups from patients with pathologically confirmed echinococcosis (cystic echinococcosis (CE), n = 69 and alveolar echinococcosis (AE), n = 38). The derived characteristic spectrum was analyzed to explain differences between echinococcosis and healthy volunteers and a principal component analysis (PCA) was applied for classification. Raman spectra revealed that high sensitivity and specificity for echinococcosis diagnosis were associated with the contents of phenylalanine and tyrosine. In addition, Raman spectroscopy analysis identified two metabolites including phenylalanine and carotenoids that could distinguish three types of serum. Orthogonal partial least squares discriminant analysis (OPLS-DA) was successfully used as a discriminative model to classify echinococcosis with the highest sensitivity and specificity of 100% and 98.6%, respectively. Combination of serum metabolomics with SERS enabled accurate screening of echinococcosis patients. The results indicate that SERS-based serum profile analysis has the potential to be a valuable tool for the early diagnosis and screening of echinococcosis.

Cystic echinococcosis (CE) is a worldwide zoonotic disease caused by the larval stage of Echinococcus granulosus. We investigated the presence of E. granulosus-specific DNA in the serum of CE patients by detecting the cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit I (nad1) mitochondrial genes. Serum and formalin-fixed paraffin embedded (FFPE) cyst tissue samples of 80 CE patients were analyzed. The extracted DNA of samples was submitted to PCR amplification of cox1 and nad1 genes, and products were sequenced and genotyped. Nineteen (23.8%; 95% CI 15.8-34.1) serum and 78 (97.5%; 95% CI 91.3-99.3) FFPE cyst tissue samples were successfully amplified with at least one gene. Echinococcus DNA was detected in the sera of 15.0% (95% CI: 8.8-24.4) and 10.0% (95% CI: 5.2-18.5) and in cyst tissue of 91.3% (95% CI: 83.0-95.7) and 83.8% (95% CI: 74.2-90.3) of 80 patients by cox1 and nad1 gene, respectively. Four genotypes of E. granulosus were distinguished in the CE patients, with predominance of genotype G1, followed by G3, G2, and G6. The finding of E. granulosus DNA in 23.8% of serum samples from CE patients confirmed that E. granulosus releases cell-free DNA into the circulatory system, but quantities may be inadequate for the diagnosis of CE. Genotype G1 predominance suggests the sheep-dog cycle as the primary route of human infection., Competing Interests: The authors have declared that no competing interests exist.

Cystic echinococcosis (CE), a zoonotic disease caused by Echinococcus granulosus at the larval stage, predominantly develops in the liver and lungs of intermediate hosts and eventually results in organ malfunction or even death. The interaction between E. granulosus and human body is incompletely understood. Exosomes are nanosized particles ubiquitously present in human body fluids. Exosomes carry biomolecules that facilitate communication between cells. To the best of our knowledge, the role of exosomes in patients with CE is not reported. Here, we isolated exosomes from the sera of patients with CE (CE-exo) and healthy donors and subjected them to liquid chromatography-tandem mass spectrometry analysis. Proteomic analysis identified 49 proteins specifically expressed in CE-exo, including 4 proteins of parasitic origin. The most valuable parasitic proteins included tubulin alpha-1C chain and histone H4. And 8 proteins were differentially regulated in CE-exo (fold change&gt;1.5), as analyzed with bioinformatic methods such as annotation and functional enrichment analyses. These findings may improve our understanding about the interaction between E. granulosus and human body, and may contribute to the diagnosis and prevention of CE.

Diagnosis of cystic echinococcosis (CE) is at present mainly based on imaging techniques. Serology has a complementary role, partly due to the small number of standardized and commercially available assays. Therefore we examined the clinical performance of the SERION ELISA classic Echinococcus IgG test. Using 10 U/ml as a cut-off point, and serum samples from 50 CE patients and 105 healthy controls, the sensitivity and specificity were 98.0% and 96.2%, respectively. If patients with other infectious diseases were used as negative controls, the specificity decreased to 76.9%, which causes poor positive predictive values. However, if results between 10 and 15 U/ml are classified as indecisive, the specificity of positive results (≥15 U/ml) increased to 92.5% without greatly affecting the sensitivity (92.0%). Using this approach in combination with imaging studies, the SERION ELISA classic Echinococcosis IgG test can be a useful aid in the diagnosis of CE.

Cystic echinococcosis (CE) is a neglected zoonotic disease with a worldwide distribution and is a major public health problem in some areas. Diagnosis of CE is mainly based on clinical symptoms, imaging and serological testing, however, improvement in serodiagnosis is still needed. This study was aimed at detecting circulating Echinococcus antigen in CE patients using a lateral flow dipstick (LFD) assay. Three types of hydatid antigens i.e. hydatid cyst fluid (HCF), native antigen B (nAgB) and recombinant antigen B (rAgB) were prepared and polyclonal rabbit antiserum was raised against each antigen. Purified IgG fractions were prepared and a portion was conjugated to gold nanoparticles. After a series of optimizations, a final antigen detection LFD assay was developed using a combination of anti-nAgB-IgG and gold-conjugated anti-HCF-IgG. Evaluation of the assay showed that 27 out of 35 (77%) serum samples from CE patients gave positive results. Meanwhile, the test showed a diagnostic specificity of 82% when tested with sera from 38 healthy individuals and 13 patients with other parasitic diseases. In conclusion, the antigen detection LFD assay seemed to be useful for diagnosis of CE and possibly for post-treatment follow-up, and merit further evaluation studies. We foresee that it may improve serodiagnosis of CE when used in tandem with an antibody detection test., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objective: To determine the effectiveness of treatment via laparoscopic approach and to present the long-term outcomes in patients with isolated renal hydatid cyst., Materials and Methods: We reviewed the data of 11 patients who underwent laparoscopic treatment for isolated renal hydatid cyst between March 2004 and January 2017. Demographic characteristics and clinical, laboratory, and radiological findings of cases were examined. Mann-Whitney U and t-test were used in statistical analysis., Results: The mean age of the patients was 36.7 years (25-51). The common complaint of the patients was lumbar pain and 9 patients had a palpable mass. It was identified that serologic evaluation of Echinococcus granulosus was positive in 5 patients. The mean cyst size was determined as 72 mm (40-150). The mean operation time was 128 minutes (120-150) while the length of hospital stay was 4 days (3-13). Perioperative major complication was not observed in any of the patients. The levels of hematocrit and creatinine were measured, 37.8 (26-43) and 0.93 (0.5-1.3), respectively, in the postoperative period, while these levels were 39 (26-45) and 0.90 (0.5-1.3) in the preoperative period and there was no statistically significant difference (P > .05). The mean follow-up time was 84 months (40-166). No relapses were observed in any patients., Conclusions: The treatment of renal hydatid cyst disease with laparoscopic approach is a safe and effective method. There was no recurrence in any of the patients in the long-term period. Further studies including a large series of patients are needed.

Background: As part of an ongoing program that aims to use early detection and timely treatment to improve the control of echinococcosis, especially in younger age groups, we undertook a series of active surveys among Qinghai-Tibetan children in the Qinghai Province of Northwestern China in 2011 and 2012. The significant outcomes that resulted from this study emphasize the need to draw attention to echinococcosis, both alveolar echinococcosis (AE) and cystic echinococcosis (CE), so that policy development is promoted and suitable avenues for control are identified in the highly endemic areas on the Tibetan Plateau., Methods: A total of 19 629 primary school students, aged 6-18 years, with a dominant Tibetan background underwent abdominal ultrasound examination, and 86.4% of the compliant students donated 2-5 ml of venous blood for serological tests. All the abnormal ultrasound results were recorded. If identified as echinococcosis, the disease lesion was assessed according to the WHO-Informal Working Group on Echinococcosis (WHO-IWGE) classification for AE and CE. Among the surveyed students, the prevalence by school was compared among geo-locations, sex and age groups. The clinical image presentations were analyzed according to lesion number, size, the location in the liver and the classification stage. Statistical significance was set at P-value < 0.05 for comparisons among groups., Results: A total of 341 students (1.7%) were identified by ultrasound as having either CE (119, 0.6%) or AE (222, 1.1%). The highest prevalence rates of childhood AE cases occurred in the Tehetu (12.1%) and Moba (11.8%) townships in Dari County. There was a high seropositive rate (37.0%) and a heterogeneous distribution of cases, with a prevalence ranged from 0 to 12.1% for AE and 0-2.9% for CE. Moreover, the seropositive rate ranged from 0.7-45.1% across different schools., Conclusions: The high prevalence of echinococcosis in Qinghai-Tibetan primary school students reflects a lack of knowledge about Echinococcus spp. transmission. The combination of systematic education for children and regularly performed anthelmintic treatment for dogs could achieve the goal of sustainable hydatidosis control.

Alveolar echinococcosis (AE) is a parasitic disease, due to Echinococcus multilocularis. Often compared to liver cancer, it develops by infiltration from its primary site to the surrounding tissue, and can then metastasize to other organs. Detection of circulating cell-free DNA (ccfDNA) is a useful analytical tool in oncology, for diagnosis, prognosis, and therapy monitoring. This study sought to investigate the presence of ccfDNA in patients with AE, and its potential usefulness for the evaluation of treatment efficiency. To achieve these aims, a quantitative PCR and a droplet digital PCR were developed to detect E. multilocularis ccfDNA. An AE animal model identified, for the first time, the presence of large quantities of ccfDNA. Samples from patients with AE (n = 31) were then analyzed twice, at diagnosis, and after three months of chemotherapy: about 25% were positive, almost always with very low concentrations of ccfDNA. These results confirmed that E. multilocularis produces ccfDNA, as solid tumors do, but detection may not yet be sufficient for AE diagnosis nor for the evaluation of treatment efficiency, due to the low levels of ccfDNA detected in patient serum., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objective: Echinococcus granulosus, the etiological agent of cystic echinococcosis (CE) in humans and livestock, is a widely distributed zoonotic pathogen tapeworm. The infection is transmitted to humans by the ingestion of E. granulosus eggs released in the feces of definitive hosts such as dogs. The larval stage of the parasite develops a slowly enlarging cyst in the visceral organs, particularly in the liver and/or lung. The aim of the present study was to evaluate the diagnostic value of an immunochromatographic test (ICT) for CE., Methods: A total of 50 sera from surgically and/or pathologically confirmed patients with CE were included in the study as the study group; the control group comprised patients who tested negative for enzyme-linked immunosorbent assay (ELISA). Sera were selected from the collection at Adnan Menderes University, Faculty of Medicine, Parasitology Laboratory, by simple random sampling. The collection included sera obtained between 2010 and 2014; antibody titers of each serum sample were determined using in-house ELISA, before storage at -20°C. The presence of E. granulosus antibody in the sera was determined using a commercially available ICT (VIRAPID® HYDATIDOSIS) kit method., Results: In the study group (E. granulosus-confirmed cases), two (4%) of the 50 sera were negative and 48 (96%) were positive with ICT. In the control group (ELISA-negative), all were negative with ICT., Conclusion: The rapid diagnostic test has been evaluated as a practical, easy-to-use method for detecting CE, and it can be used as a screening test in routine diagnosis and research.

Echinococcus granulosus is the causative agent of cystic echinococcosis (CE), a widespread parasitic zoonosis. Leucine aminopeptidases (LAPs) of the M17 peptidase family have important functions in regulating the balance of catabolism and anabolism, cell maintenance, growth and defense. In this study, we presented a bioinformatic characterization and experimentally determined the tissue distribution characteristics of E. granulosus LAP (Eg-LAP), and explored its potential value for diagnosis of CE in sheep based on indirect ELISA. Through fluorescence immunohistochemistry, we found that Eg-LAP was present in the tegument and hooks of PSCs, the whole germinal layer and adult worm parenchymatous tissue. Western blotting results revealed that the recombinant protein could be identified using E. granulosus -infected sheep serum. The diagnostic value of this recombinant protein was assessed by indirect ELISA, and compared with indirect ELISA based on hydatid fluid antigen. The sensitivity and specificity rEgLAP-ELISA were 95.8% (23/24) and 79.09% (87/110), respectively, while using hydatid fluid as antigen showed the values 41.7% (10/24) and 65.45% (72/110). This is the first report concerning leucine aminopeptidase from E. granulosus , and the results showed that Eg-LAP belong to M17 peptidase families, and that it is involved in important biological function of E. granulosus . Furthermore, rEg-LAP is appropriate for diagnosing and monitoring CE in sheep in field. Development of a rapid test using rEg-LAP to diagnose sheep CE deserves further study., Competing Interests: The authors declare no conflict of interest.

Cystic echinococcosis and toxoplasmosis are two human parasitic diseases. Butchers and slaughterhouse workers are in close contact with body fluids and tissues of ruminants. To investigate the prevalence of Toxoplasma and Echinococcus IgG antibodies in slaughterhouse workers in Northeast of Iran, Mashhad, 2016. This cross-sectional study was performed on all personnel working at the largest industrial slaughterhouse of Khorasan Province. Serum samples were taken and kept frozen until used. IgG against Echinococcus and Toxoplasma were quantified using commercial ELISA kits. A questionnaire addressing possible risk factors of infection acquisition was filled by participants. Out of 91 male participants, 58.2% were positive for toxoplasmosis, 5.5% were positive for cystic echinococcosis and 3.3% were positive for both. Except using gloves, and gown and boots, other personal protective equipment (PPE) were not completely used by all personnel; mask 38%, tool disinfectants: 12%, face and hand disinfection: 14.3%. There was a high risk of Toxoplasma in slaughterhouse workers, however, such finding was not observed in Echinococcus parasite. Importance of PPE and tool disinfectants to reduce the risk of zoonotic infections in the workplace should be emphasised.

Objective: This study was conducted to evaluate the relationship between serum levels of vitamin A and β-carotene in the liver of cattle., Methods: A total number of 150 samples were selected according to the type of parasitic infections of the liver after postmortem examination and confirmation. Parasitic lesions in the liver were subdivided into three major parasites, including Fasciola hepatica, Dicrocoelium dendriticum, and hydatid cysts. Fifty samples were obtained from cattle without any parasitic infection as a control group. Serum levels of vitamin A, β-carotene concentrations (μg/dL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activity were assayed., Results: Naturally infected cattle with D. dendriticum, F. hepatica and hydatid cyst showed lower vitamin A levels. Serum β-carotene levels were significantly decreased in all groups (p<0.05). ALT and AST activities in animals with parasitic diseases were statistically higher than in control group (p<0.01). A significant negative correlation was determined between the β-carotene, vitamin A levels and enzyme activities of the liver in all the three types of liver infections (p<0.001)., Conclusions: This study indicated that serum levels of vitamin A and β-carotene decline was present in cattle with liver parasite infection and vitamin supplements should be supplied.

Echinococcus granulosus is the causative pathogen of cystic echinococcosis, a serious disease endangering human and animal health. In this study, an immunochromatographic strip was developed based on the recombinant protein Heat shock protein 70 (HSP70) for the serological detection of E. granulosus. The protocol completes within 20min requiring no specialized equipment or chemical reagents, while specificity tests confirmed no cross-reactivity with positive serum of Fasciola hepatica, Haemonchus contortus, Peste des petits ruminants virus (PPRV) and Foot-and-mouth disease virus (FMDV). The strips remained stable after storage at 4°C for up to 8 months. Both immunochromatographic strip and ELISA tests were applied to detect E. granulosus antibody in a total of 728 serum samples obtained from slaughter houses in Zhejiang Province. Our data revealed positive rates of 2.61 and 1.65% by immunochromatographic strip and ELISA methods, respectively. The immunochromatographic strip test developed in this study provides a simple, specific and rapid method of E. granulosus antibody detection and infected sheep monitoring., (Copyright © 2017. Published by Elsevier B.V.)

Objective: To compare the performance of two novel recombinant antigens (EgP29, 2B2t) with imaging in a well-defined cohort of surgically treated cystic echinococcosis (CE) patients to determine whether serology reflects surgical cure as defined by imaging., Methods: From a cohort of 223 CE-confirmed patients of a national clinical center for echinococcosis, 36 surgically treated patients were eligible for analysis. Sera were tested by enzyme-linked immunosorbent assay (ELISA) for specific IgG and IgG4 antibodies against the EgP29 and 2B2t antigens. We used a hierarchical linear regression model to examine the course of antibody levels over time for each patient. A meta-analysis of the patient-specific estimates of the time to negativity was performed using the metan command in Stata., Results: The range of positive serological results at the beginning of post-surgical monitoring was 34-60%: 2B2t 51%, 2B2t-IgG4 34%, EgP29 60% and EgP29-IgG4 40%. The pooled estimates of time to seronegativity were as follows: 2B2t-ELISA 3.92 (3.24, 4.61) years; 2B2t-IgG4-ELISA 4.60 (3.91, 5.29) years; EgP29-ELISA 3.94 (3.50, 4.39) years; EgP29-IgG4-ELISA 2.55 (1.93, 3.18) years., Conclusion: After surgical treatment, antibodies to the recombinant antigens 2B2t and EgP29 become negative in the majority of CE-confirmed, surgically cured patients. The major drawback is the fact that only around half of the CE-confirmed, surgically treated patients were at all responsive to the test antigens, so they are of limited benefit for documenting primary cure. Equally, these antigens do not appear to be sensitive to recurrences., (© 2017 John Wiley & Sons Ltd.)

The tolerability and efficiency of specific chemotherapy with albendazole were analyzed in 18 patients with cystic echinococcosis at various sites. The most common complication of albendazole treatment was its hepatotoxic effect. Al- bendazole chemotherapy during at least its 3 cycles was observed to be highly effective in preventing recurrent echinococ- cosis in children. If there were small-sized hydatid cysts in the liver and lung, there was a reduction in their number and sizes after 4-5 albendazole treatment cycles.

Cystic echinococcosis (CE) caused by the cestode Echinococcus granulosus (E. granulosus) is a zoonotic parasitic disease. The effective immune evasion mechanisms of E. granulosus allow it to parasitize its hosts. However, the status of the innate and adaptive immune cells and their contributions to E. granulosus progression remain poorly understood. In this study, we aimed to determine the impact of E. granulosus infection on T cells, NK cell responses and TGF-β expression during the early infection phase in BALB/c mice. In E. granulosus infections, there was an increasing tendency in the percentage of CD4 + CD25 + T cells and CD4 + Foxp3 + T cells and peripheral blood TGF-β levels and relative expression of the Foxp3 gene. Moreover, there were a decreasing tendency in the percentage of NK cells and NK cell cytotoxicity and the expression of NKG2D on NK cells. The TGF-β1/Smad pathway was activated by E. granulosus in mice. Above results can be reversed by the inhibitor SB-525334 (potent activin receptor-like kinase 5 inhibitor). These results suggest that the TGF-β/Smad pathway plays an important role in changes of T-cell or NK cell responses. These results may contribute to revealing the preliminary molecular mechanisms in establishing hydatid infection., (© 2017 John Wiley & Sons Ltd.)

Cystic echinococcosis (CE) is a parasitic zoonosis especially affecting resource-poor populations in livestock raising areas. Imaging, in particular ultrasound (US), is crucial for the diagnosis, staging, and clinical management of abdominal CE in humans. Serology is a valuable complement to imaging, especially when ultrasound features of CE are absent or unclear. In rural endemic areas, where expertise in US is scant, and conventional serology techniques are unavailable due to lack of laboratory equipment, rapid diagnostic tests (RDTs) may be very useful. Several reports have described the performance of commercial and experimental RDTs in the diagnosis of CE, including a recent study by our group that compared the diagnostic performances of three commercial RDTs for the diagnosis of hepatic CE. To put RDTs for CE in context, we reviewed the available literature in English on this topic. Overall, RDTs appear to be useful in resourcepoor settings where they may replace conventional serodiagnostic tests. However, like other serodiagnostic tests, RDTs lack standardization and show unsatisfactory sensitivity and specificity. An important issue that needs to be addressed is that studies on the diagnostic performance of RDTs fail to take into account the variables known to influence results such as anatomical location and cyst stage.

Factor VII is a new coagulation factor replacement therapy. It has permitted the practice of invasive procedures which were up until recently associated with a huge risk of bleeding in patients with hemophilia with inhibitors. Our case illustrates factor replacement therapy failure in a 13-year-old child operated on for hepatic cysts associated with a pelvic cyst. Major bleeding occurred postoperatively requiring several transfusions, an increase in dosage of factor VII, and administration of a heavy dose of factor VIII as a last resort. This case highlights the possibility of failure of factor replacement therapies constituting a life-threatening situation in which alternatives are few.

Background: The larval stage of Echinococcus granulosus is the etiological agent of cystic echinococcosis (CE), which causes serious morbidity and mortality in many areas. There is no reliable method to monitor sheep CE. Here, we characterize E. granulosus glutaredoxin 1 (Eg-Grx1) and report an improved immunodiagnostic method for CE., Methods: We cloned and expressed recombinant Eg-Grx1 and generated antibodies. We analyzed the location of the protein in different parasite stages by fluorescence immunohistochemistry, detected the immunogenicity of recombinant Eg-Grx1, and developed an indirect ELISA (iELISA) for CE serodiagnosis., Results: Eg-Grx1 is a classic dithiol Grx with several GSH-binding motifs. Native Eg-Grx1 protein was distributed in the tegument of protoscoleces, the whole germinal layer, and the parenchymatous tissue of adult worms. Recombinant Eg-Grx1 exhibited good immunoreactivity to CE-infected sheep serum. An iELISA using this antigen showed specificity of 64.3 % (9/14) and sensitivity of 1:3200, and the diagnostic accordance rate was 97.9 % (47/48) compared with the results of necropsy., Conclusion: We characterized a novel Grx (Eg-Grx1) from a parasitic helminth and present a comprehensive analysis of the sequence and structure of this protein. The recombinant Eg-Grx1 protein showed good potential serodiagnostic performance, and we established an iELISA method, which may contribute to the surveillance of sheep CE in epidemic areas.

At present, various scolicidal agents have been used for inactivation of protoscoleces during hydatid cyst surgery, however, they are associated with serious adverse side effects including sclerosing colangititis (biliary tract fibrosis), liver necrosis and methaemoglobinaemia. This investigation was designed to evaluate the chemical composition and in vitro scolicidal effects of Pistacia vera (var. Fandoghi) essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. The components of the P. vera essential oil were identified by gas chromatography/mass spectroscopy (GC/MS) analysis. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (25-200μl/mL) were used for 5-30min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). In addition, forty male NIH mice were used to determine the acute and sub-acute toxicity of P. vera essential oil for 2 and 14 days, respectively. The main components of P. vera essential oil were limonene (26.21%), α-pinene (18.07%), α-thujene (9.31%) and α-terpinolene (9.28%). Findings of the present study demonstrated that the P. vera essential oil at the concentrations of 100 and 200μl/mL killed 100% protoscoleces after 10 and 5min of exposure, respectively. The LD50 values of intraperitoneal injection of the P. vera essential oil was 2.69ml/kg body weight, and the maximum nonfatal doses were 1.94ml/kg body weight. No significant difference (P>0.05) was observed in the clinical chemistry and hematological parameters following oral administrations of P. vera essential oil at the doses 0.1, 0.2, and 0.4ml/kg for 14 days. The obtained findings demonstrated new chemical composition and promising scolicidal activity of the P. vera with no significant toxicity which might be used as a natural scolicidal agent in hydatid cyst surgery., (Copyright © 2016. Published by Elsevier Masson SAS.)

Patients with cystic echinococcosis (CE) can harbour cysts for years or even decades, apparently without effect of the immune system on the metacestode. Although several immune evasion mechanisms by echinococcal cysts have been described, it is unclear whether the human leucocyte antigen (HLA) system plays a role in the susceptibility or resistance to CE in humans. HLA-G molecules are known to exert a suppressive action on dendritic cells maturation and on natural killer (NK) cells functions, therefore hampering T-cell responses and NK cytolysis. HLA-G plays an important role in immune tolerance, is involved in foetus and in allotransplant tolerance, and may be involved in tumoral and viral immune evasion. In this study, we assessed the presence and levels of soluble HLA-G (sHLA-G) in patients with CE using a commercial ELISA kit to determine whether host's HLA-G may have a role in the course of human CE., (© 2016 John Wiley & Sons Ltd.)

Objective: To investigate the levels of cytokines in the sera of mice chronically infected with the larvae of Echinococcus granulosus , and explore the mechanisms of immune regulation against parasite infection., Methods: The protoscoleces were isolated from the livers and lungs of sheep infected with E. granulosus , and then inoculated intraperitoneally to BALB/c mice (2 000 for each mouse), to establish the mouse model of E. granulosus infection. The mice in the control group were injected intraperitoneally with an equal volume of PBS. The sera of both control and infected mice were collected to test the levels of multiple cytokines by using the Cytometric Bead Array (CBA) 5 months post-infection., Results: In contrast to the control group, the multiple cysts were found in the abdominal cavity, livers and lungs of the infected mice. Moreover, the levels of inflammatory cytokines such as IL-17A, IL-6, IFN-γ, MCP-1, IL-12P70 and TNF-α in the sera of the infected mice were significantly higher than those in the control group ( t = 2.713-9.255, all P < 0.05) while the levels of anti-inflammatory cytokine IL-10 were elevated post-infection ( t = 3.936, P < 0.001)., Conclusions: Higher inflammatory cytokines in the mice chronically infected with the larvae of E. granulosus , may benefit for the limitation of parasite growth.

A standardized test for the serodiagnosis of cystic echinococcosis (CE) remains an important challenge because of the problems in specificity and sensitivity of the available commercial kits and lack of proper evaluation of antigen. Using appropriate sources of antigenic material is crucial in improvement of the serological methods such as enzyme-linked immunosorbent assay (ELISA). This study was conducted to evaluate the performance of protein named Echinococcus protoscolex calcium binding protein EPC1 for the detection of antibodies in sera from patients with CE. Expressed and purified recombinant protein EPC1 (rEPC1) was used as antigen in ELISA method. Characterization of the rEPC1 antigen was evaluated using the serum of 25 patients with both surgical and imaging confirmed CE and 25 healthy donors as negative controls. Also, a panel of sera including chronic toxoplasmosis (IgG positive), strongyloidosis, fascioliasis, toxocariasis, and kala azar were used and patients with related parasites were confirmed by medical laboratories or clinically by research centers using microscopy or specific ELISA. rEPC1 showed relatively promising performance in total IgG ELISA for the detection of antibodies in sera from the negative controls, and the cut off value 0.4 units of optical density at 490 nm was calculated for ELISA. In this study, sensitivity of 100%, specificity of 93.7, positive predictive value of 92.6%, and negative predictive value of 100% were calculated for rEPC1. On the other hand, commercial ELISA kit based on the native antigen B of Echinococcus granulosus had sensitivity of 96.2% and specificity of 96.8%. No significant difference was found for sensitivity or specificity between the rEPC1 and commercial kit. However, rEPC1 may be a valuable antigen for diagnosis of human CE., (© The American Society of Tropical Medicine and Hygiene.)

Background: Cystic echinococcosis (CE), caused by infection with Echinococcus granulosus larvae, is a potentially life-threatening disease in humans. Anaphylactic shock caused by CE is very dangerous, and is highly prevalent during surgery. Dexamethasone (DEX) is used clinically before operations to prevent allergic reactions; Regulatory T cells (Treg cells) are believed to be associated with negative immune response, which play an important role in alleviating allergic reactions. However, the association of Treg cells with DEX remains unknown., Methods: In this study, C57BL/6 mice were divided into uninfected group, untreated group and DEX group which were inoculated with protoscoleces from E. granulosus and sensitized using a cyst fluid suspension to induce anaphylactic shock. In addition, the mice in DEX group were treated with 10 mg/kg DEX by intraperitoneal injection 30 min before being sensitized., Results: It was found that 93.75 % of all sensitized mice experienced allergic symptoms. The levels of IgE, IgE/IgG, and IgE/IgG1 were significantly higher in both untreated group and DEX group. The proportion of CD4 + CD25 + FOXP3 + Treg cells relative to CD4+ Treg cells, and the levels of interleukin-10 (IL-10) and tumor growth factor-β (TGF-β1) were significantly higher in DEX group. The level of IL-13 was significantly higher in the sensitized mice than in the other groups. These cells may play a key role in alleviating the immune response in CE-induced anaphylactic shock., Conclusions: The protective effect of DEX may be due to Treg cell upregulating IL-10 and TGF-β1 levels, and inhibiting helper T cell 2 cytokines.

Objectives: To study the hematological findings of dogs infected with echinococcosis and the possibility of using in vitro reared Echinococcus granulosus excretory-secretory antigen (ES-Ag) as a reagent for serological diagnosis of canine echinococcosis., Methods: Eight dogs were infected orally with protoscoleces, extracted from ovine fertile hydatid cysts. Two additional dogs were infected with Cysticercus tenuicollis, obtained from infested sheep. The hematological parameters of dogs were determined with the ADVIA 2120i automatic hematology analyzer with a blood smear staining module. Adult E. granulosus and/or Taenia hydatigena that were collected from small pieces of the open gut and the larval cestodes that were extracted from infested sheep during slaughter were cultured in an incomplete RPMI-1640 medium. The parasite-ES-Ag-containing supernatant was used as an antigen in enzyme linked immunosorbent assay (ELISA) to detect antibodies in the sera of infected dogs., Results: A significant increase in hemoglobin concentration and erythrocyte count was found during the infection, as well as an increasing proportion of lymphocytes and segmented neutrophils, accompanied by a significant reduction of the leukocyte count and a growth of both the absolute and the relative eosinophil count. ELISA found a strengthening antigenicity of echinococcus preparations during infection. This property was more pronounced in the protoscoleces ES-Ag, compared to the eponymous antigen of an adult parasite. The latter gave specificity to ELISA, which allowed differentiating it from the similar antigen of the closely related tapeworm T. hydatigena., Conclusion: In vitro reared adult E. granulosus ES-Ag can be used as an antigen in the serological diagnosis of canine echinococcosis. Hematological parameters and serological results have predictive value in the screening of dogs for echinococcosis; however, in some individuals, they may reflect the state of resistance to invasion.

Human cystic echinococcosis (CE), caused by Echinococcus granulosus, is one of the most important and widespread parasitic zoonoses. One of the problems that can be encountered after treating CE patients is the risk of post-surgical relapses or treatment failure, thus a long-term clinical and serological follow-up is required to evaluate the success or failure of therapy. In the present study immunological markers have been identified to indicate the effectiveness of pharmacological and surgical treatments. The relationship between serum cytokine levels and the outcome of chemotherapy and surgery was evaluated in 50 patients with CE. Serum interleukin (IL)-4, IL-10 and interferon-gamma (IFN-γ) concentrations were determined by enzyme-linked immunosorbent assay (ELISA) before and after pharmacological and surgical treatment. Serum cytokine levels of IL-4, IL-10 and IFN-γ were elevated in a significant proportion of patients during the active stage of disease. IL-4, IL-10 and IFN-γ were measurable in 41 (82%), 37 (74%) and 25 (50%) patients before the treatment. Clinical and radiological assessment of patients 2 years after pharmacological treatment has shown that 48 of 50 patients responded to treatment. IL-4 and IL-10 levels were decreased significantly (P< 0.05) in these patients. Conversely, patients who did not respond showed high levels of IL-4 and IL-10 and undetectable levels of IFN-γ. Hence these results suggest that serum IL-4 and IL-10 detection may be useful in the follow-up of patients with CE.