Human Hydatid Cyst Research Articles

Use of the EmsB microsatellite-based next generation sequencing for genotyping of Echinococcus granulosus sensu lato in hydatid cyst tissue samples from animals and humans.

Echinococcus granulosus (Batsch, 1786), a cestode of the Teniidae family, causes human cystic echinococcosis (CE) also known as hydatid disease. Echinococcus granulosus sensu lato includes the G1, G3, G4, G5, G6/7 and G8/10 genotypes which are known to cause human CE. This study aimed to differentiate genotypes of E. granulosus s.l. complex by employing EmsB, a tandemly repeated multilocus microsatellite, using next-generation sequencing (MIC-NGS). Human and animal histopathology-confirmed hydatid cyst tissue samples and reference DNA samples of E. granulosus G1, G3, G4, G5, G6/7 and G10 underwent MIC-NGS assay with custom primers amplifying a 151 bp EmsB DNA fragment. NGS data were analysed using online Galaxy analysis pipeline, a phylogenetic tree was constructed by MEGA software, and haplotype networking was performed with PopArt 1.7. All sixty samples (49 from animals and 11 from humans) included were successfully identified and genotyped with a 100 % success rate. The study showed improved discrimination power to distinguish all study samples including closely related E. granulosus s.s. genotypes G1-G3. The maximum likelihood tree reaffirmed the monophyly of E. granulosus s.l. The median-joining haplotype networking revealed 12 distinct haplotypes. In conclusion, MIC-NGS assay was shown to be sensitive, specific and simple to apply to clinical samples offering a powerful discriminatory tool for the genotyping of E. granulosus s.l.

Diagnosis of Human Cystic Echinococcosis by detecting Antigen B in Serum and Urine using Nanomagnetic Beads-Enzyme Linked Immunosorbent Assay

Abstract Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the larval stage; hydatid cyst, of E. granulosus. The disease is usually asymptomatic, and mostly presents by pressure symptoms according to its location. The diagnosis of CE is currently based on imaging techniques, which may not be available in some areas and antibody detection in serum, with several known drawbacks. Conversely, antigen detection has proven its efficacy in serodiagnosis of CE. The study aimed to test the diagnostic efficacy of Nanomagnetic beads-enzyme linked immunosorbent assay (NMB-sandwich ELISA) for diagnosis of CE by detection of E. granulosus AgB in serum and urine samples in comparison to conventional sandwich ELISA. The study included 42 individuals classified into 3 groups; CE infection group (Gp I), other parasitic infections group (Gp II) and healthy control group (Gp III). E. granulosus AgB was prepared from human hydatid cysts and used to produce anti- E. granulosus AgB-IgG-pAbs in rabbit which was used to detect AgB level in serum and urine samples by sandwich ELISA and NMB-sandwich ELISA. Using sandwich ELISA, AgB was detected in serum samples of 88.9% of CE patients with sensitivity and specificity of 88.9% and 91.7%, respectively. In urine samples, sandwich ELISA detected AgB in 83.3% of CE patients and the sensitivity and specificity were 83.3% and 87.5%, respectively. NMB-sandwich ELISA detected AgB in serum samples of 94.4% of CE patients with a sensitivity and specificity of 94.4% and 95.8%, respectively, whereas in urine samples it was detected in 88.9% of CE cases and the sensitivity and specificity were 88.9% and 95.8%, respectively. There was no statistically significant difference (p > 0.05) between the use of serum and urine samples for detection of AgB when examined by either sandwich ELISA or NMB- sandwich ELISA. The best test accuracy was obtained with NMB-sandwich ELISA detecting AgB in serum samples (95.2%), whereas, it showed a slightly lower accuracy of 92.9% when using urine samples. It was concluded that NMB-sandwich ELISA is efficient for detection of AgB in serum samples. Moreover, urine samples might be a good alternative to serum samples for the detection of AgB showing reasonable sensitivity, specificity and accuracy.

Complete characterization of NADH dehydrogenase subunit 1 gene in human hydatid cysts

يمتلك طفيلي الاكياس العدرية سلالاتين متشابهتين (G1 و G3) ونظرا للدور البارز الذي تلعبه عائلة جينات نازعات الهيدروجين في التميز بين هاتين السلالاتين، لذا هدفت الدراسة الحالية الى اجراء التوصيف الجزيئي الكامل لاحد ابرز انواع هذه العائلة وهو الجين NAD1 (من شفرة البدء الى الي شفرة التوقف) للاكياس العدرية المعزولة جراحيا من اكباد المرضى (ولمحافظتي البصرة وميسان) في مستشفى الصدر التعليمي في محافظة البصرة، العراق. عزلت الرؤيسات الاولية من الاكياس تحت ظروف معقمة واستخدمت في استخلاص الحامض النووي منقوص الاوكسجين. اذ ضخم التتابع الكامل للجين في عينات الدراسة وانجز التتابع (السكونس) لهما باستخدام بادئات متخصصه. اظهرت نتائج تحليل السكونس المتكرر Consensus sequence امتلاك كل من عينات البصرة و ميسان 1038 زوجا من القواعد وتقع شفرة البدء GTG في الموقع 87-89 وشفرة التوقف TAA في الموقع 978-980. كما اظهرت استدلالات الاحماض الامينية الى وجود 297 حامض اميني لبروتينات عينات البصرة وميسان. فضلا عن ذلك بينت تحاليل المعلومات الحياتية احتواء هذه البروتينات على الحامض الاميني Isoleucine في الموقع 188 (I188) وهو موجود فقط في سلالة الاغنام G1. وان النموذج التركيبي لبروتينات عينات البصرة وميسان اظهر تشابها كبيرا و يقع I188 في الحلزون 10 (α10). بينت الدراسة ولاول مرة على مستوى العراق التوصيف الكامل للجين NAD1 والعائد الى سلالة الاغنام G1 المسببه لمرض الاكياس العدرية البشري.

Human and camel cystic echinococcosis – a polyclonal antibody-based sandwich ELISA for its serodiagnosis with molecular identification

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.

Identification and genotyping of Echinococcus granulosus from human clinical samples in Guilan province, north of Iran

Cystic echinococcosis (CE) is a significant health problem in both human and veterinary medicine. It is caused by the tapeworm Echinococcus granulosus (E. granulosus). The objective of this study was to investigate molecular diversity of E. granulosus from the paraffin-embedded human (FFPE) tissue samples using sequencing of mitochondrial genes. Thirty-five FFPE tissue samples were collected from different regions of Guilan province, north of Iran. Demographic data were recorded using a questionnaire. Five sections (1 mm) of the tissue were prepared and deparaffined using xylene and ethanol methods. Molecular analysis was performed using the Nad1 and Cox1 genes using PCR and DNA sequencing. Totally, 25 cases (71.43%) were women and 10 cases (28.57%) were men. The most affected age group was 21–30 yr old. The most of cysts were isolated from the liver (n = 19; 54.29%) and others in the lung (n = 16; 45.71%). The Cox1 and Nad1 genes were successfully amplified in 16 (45.71%) and 12 (34.28%) DNA samples from FFPE tissue. Sequencing analysis revealed that all samples were E. granulosus sensu stricto complex (G1 and G3). In this study, E. granulosus sensu stricto complex G1 and G3 were identified in human hydatid cysts and showed the presence of sheep/dog cycle in human infection. This finding confirmed and completed previous studies on the geospatial distribution of E. granulosus sensu stricto complex G1 and G3 in the southern and coastal areas of the Caspian Sea region.

Molecular discrimination of G1 and G3 genotypes of Echinococcus granulosus sensu stricto obtained from human, cattle, and sheep using the mitochondrial NADH dehydrogenase subunit 5 marker

Cystic echinococcosis (CE) is a common zoonotic disease caused by the larval form of Echinococcus granulosus sensu lato. This study determined the genotype and haplotype differences using the NADH dehydrogenase subunit 5 gene in hydatid cyst samples. Human (n = 12), cattle (n = 28), and sheep (n = 31) hydatid cyst isolates were included. Seventy-one genomic DNA samples were successfully extracted, and a 759 bp mitochondrial NADH dehydrogenase subunit 5 gene fragment was amplified by PCR. Following the sequence analysis, E. granulosus sensu stricto isolates were identified as G1 (n = 61) and G3 (n = 10). A total of 23 haplotypes were obtained from the 71 E. granulosus s.s. G1 and G3 samples. The main haplotype was Hap01 (60.56 %), which consisted of the G1 genotype. The second largest haplotype was Hap04, which consisted entirely of the G3 genotype. Hap14 acted as a bridge between the G1 and G3 genotypes. This study identifies G1 as the dominant genotype in humans and farm animals in Turkey. High haplotype and nucleotide diversity in genotypes were observed. Additionally, this is the first report on the phylogeography and gene flow models of the E. granulosus s.s. population in Turkey using the NADH dehydrogenase subunit 5 gene, the best marker distinguishing between G1 and G3 genotypes.

Molecular Epidemiology of Cystic Echinococcosis in Rural Baluchistan, Pakistan: A Cross-Sectional Study.

Cystic echinococcosis (CE), or hydatid cyst disease (HCD), is a zoonosis of significant importance caused by the cestode of Echinococcus granulosus sensu lato (s. l.) that affects mainly nomadic populations and has substantial economic consequences. Due to the 76% rural and nomadic population, Baluchistan is a highly endemic region in Pakistan for CE; however, it has not yet been investigated for CE. For this purpose, this study was carried out to investigate the molecular epidemiology of CE in this region. In total, 23 human hydatid cyst samples were collected from tertiary health care units in Baluchistan and processed for DNA extraction, which was then followed by sequencing of the cox1 mitochondrial gene of all 23 collected samples, genotyping, and phylogenetic and haplotype analysis. Most subjects were livestock owners (39.13%) in rural settings (73.91%). Most patients (73.19%) were pet owners (dogs) and used water from open sources for drinking. The liver was the most affected organ (52.17%), followed by the lungs (17.39%). Sequence analysis based on the cox1 gene revealed that EG genotype 1 (G1) was the most prevalent (56.52%), followed by G3 (34.78%), while some samples (8.7%) were identified as the Echinococcus canadensis (G6/7) genotype. A total of five haplotypes were detected with high haplotype diversity (0.80) and low nucleotide diversity (0.033). Phylogenetic analysis revealed two diverse sub-clades, each of G1 and G3 isolates from Baluchistan, that were evolutionarily related to previously reported G1 and G3 isolates from Pakistan and China. On the other hand, the G6/7 isolates of this study were evolutionarily identical to the already reported G6/7 isolates from Pakistan, Turkey, and Kazakhstan. This study concludes that diverse G1 and G3 EG isolates are present in this part of Pakistan, while the G6/G7 genotype was reported for the first time from Baluchistan.

Histopathological and immunological study of rats liver hydatid cysts isolated from human, sheep, goat and cows.

In endemic places, liver hydatidosis is a life-threatening health issue. Many consequences such as hepatomegaly, infiltration with inflammatory cells and histopathological changes might arise as a result of liver hydatidosis. This study aimed to look into pathogenic changes in the livers of the rats that experimentally infected with hydatid cysts. These hydatid cysts were isolated from naturally infected humans, sheep, goats and cows. Liver hydatid cysts were collected from the main abattoir of Al-Muthanna province while human hydatid cysts were collected from Al-Hussein Teaching Hospital in Al-Muthanna province. The hydatid cysts were grossly and histology examined for inspection of hydatid cysts. The in vivo experiments were done by injection of hydatid protoscoleces or sand (fluid) in rats intraperitoneally. The results showed that the gross signs were same in all infected livers which range from paleness, hepatomegaly, hemorrhage and calcification. The hydatid cysts isolated from sheep and goat livers were highly fertile compared to others isolated from humans and cows. Injection of hydatid protoscoleces and hydatid fluid isolated from sheep and goat livers in rats induced the highest immune response compared to that isolated from humans and cows. The liver sections of rats that were injected with human, goat and sheep hydatid protoscoleces and fluids showed hyperplasia in the bile duct, aggregation nonnuclear cells with congested blood vessels. While liver sections of rats were received goat hydatid fluid, cow hydatid protoscoleces and hydatid fluid showed normal liver tissue. These findings suggested that the immunogenicity of hydatid materials is different according to the host and the component of hydatid cysts.

The comparison between AgB-ELISA and a new method of Nano-ELISA for diagnosis of hydatidosis in sheep

Hydatidosis is one of the most important zoonotic diseases, and it is transmitted via dogs to intermediate hosts such as humans and domestic animals including sheep and cattle. Epidemiological studies and genetic investigations indicate that the sheep strain is the most common species causing hydatid cysts in humans. The prevalence and incidence of this disease are increasing. According to surveys, economic losses due to this parasite in intermediate hosts are significant. In this survey, 25 serum samples were obtained from newborn lambs as negative control and obtained 25 serum samples from infected sheep to hydatidosis as the positive control. Antigen B isolated from hydatid cysts fluid was used for designing ELISA methods. Using Antigen B in ELISA design for hydatidosis diagnosis has attracted researchers in recent years. During this study, an Iranian native B antigen was used to design the specific detection of hydatidosis in sheep using a specific ELISA technique. The first method used the anti-Sheep conjugate (SIGMA, at 1:3000 dilution), and the second method used gold nanoparticles in combination with anti-sheep conjugate. According to the results, sensitivity and specificity in sheep of the AgB-ELISA method were both 92% and for the Nano-ELISA with Gold Nanoparticles 100% and 96%, respectively. Moreover, results indicated that using Antigen B in ELISA design is valuable but specificity and sensitivity will increase significantly, especially in lower titer, when gold nanoparticles with anti-sheep conjugate are used.

CHARACTERIZATION OF PREVALENT ECHINOCOCCUS GRANULOSUS GENOTYPES THROUGH MODIFIED PCR-RFLP TECHNIQUE

Background:
 Cystic echinococcosis is a zoonotic, neglected tropical disease caused by various genotypes of cestode parasite Echinococcus granulosus. Although, echinococcosis causes enormous financial and health impacts, no gold standard diagnostic technique is available. Genetic characterization of these prevalent cestode at species level is essential for disease management and appropriate control measures.
 Objective:
 We aimed to investigate the genetic diversity of echinococcus granulosus using a modified Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) based assay in infected population.
 Methodology:
 A total of 18 human hydatid cyst samples were collected from various hospitals of Southern Punjab and Islamabad Capital Territory region of Pakistan. Extracted DNA was used for PCR amplification of mitochondrial NADH dehydrogenase subunit 1 (Nad1) gene followed by sequencing and phylogenetic analysis using Molecular evolutionary Genetic Analysis (MEGA) Software. The entire sequences were fed into NEBcutter V2.0 to select a single restriction enzyme followed by invitro confirmation through PCR-RFLP.
 Results:
 Amplification on the Nad1 gene was observed in 100% of the samples processed. The Basic Local Alignment Tool (BLAST) and phylogenetic tree analysis revealed 83.3% E. granulosus (G1-G3 genotypes), 11.1% E. multilocularis and 5.6% E. Canadensis (G6 genotype). The use of the BfaI enzyme in PCR-RFLP analysis revealed that all of the 18 samples were assigned consecutive genotypes as observed in the sequencing.
 Conclusion:
 The current study concluded that the BfaI enzyme could be used for the genotypic analysis of echinococcosis in developing and frequently affected countries. It will be a cost-effective and easy technique compared to sequencing, which will aid in developing novel therapeutic and control strategies for the parasite.

Interleukin 17 Cytokine Profiles in Patients with Cystic Echinococcosis in Babylon Province, Iraq.

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.

Expression and production of protoscolex recombinant P29 protein and its serological evaluation for diagnosis of human hydatidosis.

Cystic Echinococcosis/Hydatidosis considered as one of the most important parasitic diseases in humans and animals across the world. The goal of the present study was to determine a native antigen with an acceptable sensitivity and specificity to be used in the human hydatid cyst diagnostic methods. In the present study, recombinant P29 antigen was used to detect the antibodies in the serum of patients with hydatid cysts of the liver. In fact, purified recombinant P29 protein is used as an antigen in ELISA. In order to evaluate the recombinant P29 protein for diagnostic ELISA, 25 serums obtained from people harboring the hydatid cysts were tested. The result of the gene expression on a 12% SDS-PAGE gel showed a band with a length of 28 KD. Also, 28KDa band was observed through the reaction of recombinant P29 protein with Anti T7-tag monoclonal antibody in the western blotting method. This protein showed satisfactory results in detecting the hydatid cyst antibodies in the serum of patients having hydatid cysts. Twenty two of 25 hydatidosis serums positively reacted in ELISA using with P29 protein, indicating in 92% of ELISA sensitivity, 95% of specificity, 95.83% of positive predictive value, and 90.42% of negative predictive value for recombinant P29 protein. Whereas the produced recombinant protein P29 showed promising results in the diagnosis of hydatidosis but of more research needs to be done to reach a more accurate conclusion.

Echinococcus granulosus sensu stricto G1 is the predominant genotype in human and livestock isolates from Turkey and Iran, based on mitochondrial nad5 gene differentiation

BackgroundDifferent genotypes of Echinococcus granulosus sensu stricto (s.s.) isolated from livestock and humans have been identified based on cox1 and nad1 genomic fragments. The present study was performed to differentiate the G1/G3 genotypes of Echinococcus granulosus (s.s.) isolated from humans and livestock (sheep and cattle) from Azerbaijan in northwestern Iran, Fars Province in southern Iran, and Van province in Eastern Turkey, using the nad5 gene fragment as a suitable marker to distinguish these two genotypes.MethodsA total of 60 pathologically confirmed human hydatid cysts and 90 hydatid cyst samples from livestock were collected from Turkey and Iran. PCR was performed on all of the samples, targeting the nad5 gene. Based on PCR product quality, host type, and the geographical area where the samples were obtained, 36 of the samples were sequenced and were used in the phylogenetic analysis.ResultsOut of 36 evaluated samples, 26 (72.2%) samples belonged to G1, and 10 (27.8%) samples belonged to the G3 genotype. Out of 21 samples from Turkey, 16 (76.2%) were G1 and 5 (23.8%) were G3, while out of 15 samples from Iran, 10 (66.7%) were G1 and 5 (33.3%) were the G3 genotype. None of the samples isolated from humans in Iran or from sheep in Turkey were G3. Overall, between the two countries, 18.18% of E. granulosus isolates in cattle, 41.66% of isolates in sheep, and 23.07% of human samples were identified as G3, and the others as the G1 genotype. The G3 genotype was not detected in human samples from Iran or sheep samples from Turkey.ConclusionThe findings of the study revealed that the G1 genotype of E. granulosus s.s. is the predominant genotype in humans and livestock, both in Turkey and Iran. The ratio of the E. granulosus s.s. G1 to G3 genotype was 3.2 in Turkey and 2 in Iran. The study also further confirmed that the nad5 gene properly differentiated the G1/G3 isolates of E. granulosus from both humans and livestock.Graphical

Employing NADH Dehydrogenase Subunit 1 in the Determination of Echinococcus granulosus Strain in Sheep, Cattle and Human in Thi-Qar Province, Iraq

Echinococcosis is a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus. This disease is an important public health and a significant economic issue in Iraq, where the lungs and livers are the popular places of infection. The aim of the current study focused on using the molecular techniques in the detection of an E. granulosus strain that causes cystic echinococcosis to human, sheep and cattle in Thi-Qar province, Iraq. In the current study, thirty isolates of E. granulosus were collected from 10 human hydatid cysts through surgery done at Al-Hussein Imam Teaching Hospital in Thi-Qar province and 10 sheep with 10 cattle hydatid cysts were obtained from the slaughterhouse in Thi-Qar province, Southern of Iraq to identify strains of E. granulosus which infect human and other intermediate hosts (sheep and cattle). The molecular study was carried out on the isolates and a specific primer set for the mitochondrial dehydrogenase NADH subunit 1 (NAD1) gene was used. This primer set was amplified 400 bp of the NAD1 gene in all selected isolates. The PCR products for the twelve selected isolates of E. granulosus (4 isolates per intermediate host) were sequenced and the results for these twelve isolates showed that all sequenced isolates, except one isolate Eg_5, belonged to the sheep strain G1 and a slight genetic diversity was observed with the reference sequences of the strain G1. The exception was in the isolate Eg_5 isolated from a cattle liver, which was similar to the buffalo strain G3. This study concludes that the common E. granulosus strain in Thi-Qar province is G1.

Seroprevalence of hydatid cyst and related risk factors in humans referred to urban health centers of Zahedan city, southeast Iran

IntroductionCystic echinococcosis (CE) or hydatidosis is one of the most common diseases between humans and livestock that humans are considered as an accidental host. This disease has spread globally, especially in the Mediterranean region, including Iran. To the best of our knowledge, no specific research has been conducted to investigate the epidemiological status of this disease in Zahedan, Sistan and Baluchestan Province, Iran. Therefore, this study was performed to achieve that aim for the first time. MethodsA cross-sectional study was performed to evaluate the seroprevalence of hydatid cyst among patients (n = 773) referring to Zahedan health centers within 2019–2020. Zahedan health centers were classified into five geographical regions. The sampling process from each area was conducted using a simple randomized cluster method. The correlation between seropositivity and risk factors was evaluated using the Chi-square test. ResultsThe seroprevalence of hydatid cyst in Zahedan city was estimated at 3.6% (28 out of 773 samples). It was found that females were affected by this infection more than males, and the highest hydatid cyst seroprevalence rate was revealed in the age group of 31–50 years and the northeastern area of Zahedan. Although age, gender, vegetable consumption, soil contact had no significant influence on the risk of CE infection (P-value>0.05), the relationship of keeping dogs in the household and the presence of stray dogs in a living environment with seropositivity to CE was significant (P-value<0.05). ConclusionThe findings of this study showed that human hydatid cyst was common in Zahedan. Therefore, the implementation of preventive and hygienic principles, especially the presence of dogs in the household or stray dogs in the environment, should be a priority. Moreover, the various epidemiological aspects of this disease in Zahedan city should be taken into consideration.

Human hydatid cyst fluid-induced therapeutic anti-cancer immune responses via NK1.1+ cell activation in mice.

Echinococcus granulosus is a cestode parasite which causes cystic echinococcosis disease. Previously we observed that vaccination with E. granulosus antigens from human hydatid cyst fluid (HCF) significantly inhibits colon cancer growth. In the present work, we evaluate the anti-tumor immune response induced by human HCF against LL/2 lung cancer in mice. HCF vaccination protected from tumor growth, both in prophylactic and therapeutic settings, and significantly increased mouse survival compared to control mice. Considering that tumor-associated carbohydrate antigens are expressed in E. granulosus, we oxidized terminal carbohydrates in HCF with sodium periodate. This treatment abrogates the anti-tumor activity induced by HCF vaccination. We found that HCF vaccination-induced IgG antibodies that recognize LL/2 tumor cells by flow cytometry. An antigen-specific immune response is induced with HCF vaccination in the tumor-draining lymph nodes and spleen characterized by the production of IL-5 and, in less extent, IFNɣ. In the tumor microenvironment, we found that NK1.1 positive cells from HCF-treated mice showed higher expression of CD69 than control mice ones, indicating a higher level of activation. When we depleted these cells by administrating the NK-specific antibody NK1.1, a significantly decreased survival was observed in HCF-induced mice, suggesting that NK1.1+ cells mediate the anti-tumor protection induced by HCF. These results suggest that HCF can evoke an integrated anti-tumor immune response involving both, the innate and adaptive components, and provide novel insights into the understanding of the intricate relationship between HCF vaccination and tumor growth.

Seroprevalence of human hydatid cyst: a cross sectional study in a rural areas of Zahedan, southeastern Iran.

Hydatidosis or cystic echinococcosis (CE) is one of the most common zoonosis diseases. Iran is one of the endemic regions in terms of this disease. For the first time, the present study was conducted to determine the seroprevalence of hydatid cyst in Zahedan rural areas due to the importance of human CE and lack of information in this region. The present study was performed on 551 people referred to seven rural health centers in Zahedan during 2019-2020. Serum samples were collected and analyzed by indirect ELISA method using recombinant antigen B subunit B8/1. Results were analyzed by SPSS (version 22) software and Chi-square test. The CE seroprevalence was 4%. The most positive cases were in the age group of 10-30 years. The highest infection was reported in homemakers. A significant relationship (P-value<0.05) was only reported between the seropositivity to hydatid cyst and the presence of dogs in the environment. The present study's findings indicated human hydatid cyst in rural areas of Zahedan is a health problem; moreover, the control and prevention principles and analysis of various epidemiological aspects of this disease should be considered.

Mutation occurrence in nad1 and cox1 genes of human strain hydatid cysts isolated from infected patients

Mutation occurrence in nad1 and cox1 genes of human strain hydatid cysts isolated from infected patients

Animal hydatid cyst genotypes as a potential substitute for human hydatid cyst as a source of antigen for diagnosis of zoonotichydatidosis.

The Diagnosis of hydatidosis is still an unsolved issue due to difficulties in obtaining of patient's hydatid cyst appropriate for antigen extraction. This study evaluated the suitability of HC protoscolices somatic antigens (HCPsS-Ag) fractions from animal origin to substitute that extracted from HC of patients in diagnosis of hydatidosis using enzyme-linked immunoelectro-transfer blot and Enzyme-linked immunosorbent assay (ELISA). Eight fractions in HC-G6 from patients react specifically versus HC-G6 infected patient's sera. Five of them (28, 32, 38, 59 and 89 Kilo Dalton (KDa) and two of them (28KDa and 45KDa) reacted versus HC-G1 and HC-G4 infected sheep and equine sera, respectively. Six fractions in HCPsS-Ag-G1 of sheep react versus HC-G1 sheep infected sera, four (28, 32, 52 and 58 KDa) and two of them reacted versus HC-G6 and HC-G4 infected patient and equine sera, respectively. Two fractions only in HCPsS-Ag-G4 of equine reacted versus infected human and sheep sera. This fraction displayed the same degree of ELISA value versus different infected sera with a significantly perfect classification for kappa agreement and non-statistically significant difference (p ≥ 0.05) for ELISA Optical density values of the positive samples without cross-reaction versus other parasites antibodies in sera. HCPsS-Ag from HC genotypes that developed in humans and animals as HC-G6 and HC-G1 can substitute each other for diagnosis of infection than antigens extracted from non-zoonotic HC-G4. The fraction at 28KDa is the only fraction that can be extracted from any animals HC and used in diagnosis of zoonotic hydatidosis.

Ultrastructure of Human Fertile Hydatid Cysts Using a Scanning Electron Microscope (SEM)

Introduction: Echinococcosis and hydatidosis caused by the metacestode of Echinococcus granulosus are among the most important zoonotic diseases in the world. This study aims to study the ultrastructure of fertile hydatid cysts that infect humans using a scanning electron microscope (SEM). Materials and Methods: Twenty samples of human fertile hydatid cysts were collected from the human liver and lung after performing surgery operations and examined with an SEM. Results: The results of the electron microscopy with different magnifications revealed that the laminated layer (LL) consists of sheets that appeared more compact and aligned. The brood capsules appeared, consisting of a net of finger-shaped structures that emerged from bulges of various sizes and shapes. Conclusion: Under a transmission electron microscope, it was found that the LL had a coherent and flexible structure, settling on a three-dimensional microscopic network of hydrophilic fibers, with high humidity. These fibers were arranged irregularly and had a diameter of about 10 nm; therefore, the fibers adjacent to the germinal layer (GL) were possibly attached to microtriches of tegument, which reached a thickness of 1 mm in the LL.