Seroprevalence of Toxocara spp. in children (3-13 years old) in Zahedan, Southeast of Iran.

Child Toxocariasis (CT) is one of the most important helminthic parasitic diseases in Iran. The current study was aimed to determine the seroprevalence of toxocariasis in children in Zahedan, southeast of Iran. In this cross-sectional study, serum samples of 373 children aged 3-13 years old referred to health centers in Zahedan were collected randomly with criteria based on the parameters that were used in earlier studies and examined for anti-Toxocara antibody using a commercial ELISA kit. Of the 373 recruited subjects, 206 (55.2%) were male and 167 (44.8%) were female. Most of the subjects (36.5%) were in the age group 3-4 years old. Anti-Toxocara antibodies were detected in the sera of five out of 373 individuals, corresponding to a seroprevalence rate of 1.3%. A significant correlation was found between the incidence of disease and eosinophilia, a history of contact with dogs or cats, as well as the consumption of raw vegetables and drinking of contaminated water. There was no significant correlation between prevalence and gender or age. The finding of this study revealed that the prevalent CT is relatively low in Zahedan region. The rate of CT infection in the southeast of Iran was lower than the rate in other parts of Iran.

Clinical Features, Diagnosis and Management of Patients with Suspicion of Fascioliasis in Kohgiluyeh and Boyer-Ahmad Province, Southwestern Iran.

BACKGROUND: In the current study, we described the epidemiological features, clinical presentation, diagnosis and management of patients with suspicion of fascioliasis in Kohgiluyeh and Boyer-Ahmad Province in southwest of Iran. METHODS: Overall, 56 patients with suspicion of fascioliasis, based on their clinical signs and symptoms that referred to Clinic of Internal Medicine in Yasuj city, from 2014 to 2016 were enrolled. Demographic data, history of eating aquatic local plants, the chief complains, and laboratory findings were recorded for each patient. Stool samples were obtained from each case for detection of Fasciola eggs. Moreover, blood samples were taken from each patient and evaluated for detection of anti-Fasciola antibodies by an indirect ELISA. Patients who defined as having fascioliasis were treated with triclabendazole and were followed for at least three months for clinical improvement. RESULTS: Serological test was positive in 5 patients. Of these 5 cases, three cases had a history of ingesting raw aquatic vegetables. The main clinical signs and symptoms in positive cases were; abdominal pain (60%), epigastric pain (40%), anemia (60%), and dermal pruritus (20%). Hypereosinophilia was seen in all of 5 positive cases. No Fasciola egg was found in stool specimens of any of the patients. The fascioliasis cases were treated by triclabendazole and clinical symptoms disappeared in all of 5 cases. CONCLUSION: Our observation further confirmed Yasuj district as a human endemic area for fascioliasis in Iran. The study also highlighted the importance of clinical features together with eosinophilia, as key parameters, in the diagnosis of human fascioliasis. Clinicians need to be aware of this disease and should keep in mind fascioliasis when hypereosinophilia present in patients in such endemic areas.

Molecular Evaluation of a Case of Fasciola hepatica in Wild Boar in Southwestern Iran: A Case Report.

Wild boars may be infected with several zoonotic parasitic infections including Fasciola spp. We reported a case of Fasciola infection in a wild boar in Bushehr Province in southwestern Iran. The sample was isolated from the liver of a hunted wild boar. A few of adult worms were fixed and stained. DNA was extracted from apical and lateral parts of the worms and PCR amplified, targeting NADH dehydrogenase subunit 1 (nad1) and cytochrome C oxidase subunit 1 (cox1) mitochondrion genes. Although the worm was quite long and looked much similar to F. gigantica, sequencing and analysis of PCR products of nad1 and cox1 genes revealed that the isolate has the most similarity with F. hepatica. This is the first report of molecular evaluation of Fasciola spp. from wild boar in Iran.

Diagnostic accuracy of urinary latex agglutination test (KAtex) for the diagnosis of visceral leishmaniasis: A meta-analysis.

Latex agglutination test (KAtex) has been used in the last two decades for the diagnosis of visceral leishmaniasis (VL) in different VL-endemic areas. Here, we present a meta-analysis of studies which evaluated the KAtex for the diagnosis of VL to find out its overall diagnostic performance. A database search was performed on PubMed, Scopus, ISI Web of Science, Iranmedex and Google Scholar. The search of databases found 57 papers, of which 17 articles fulfilled our eligibility criteria. Meta-analysis of diagnostic accuracy (MADA) and Hierarchical Summary Receiver Operating Curve (HSROC) packages were used to do the meta-analysis and to obtain pooled estimates of sensitivity and specificity. Fixed effect bivariate analysis was conducted, using Mantel-Haenszel estimator, to measure the performance and diagnosis odds ratio (DOR) of the test. Heterogeneity of the test results was assessed by Chi-squared test. The sensitivity of individual studies ranged from 39.8 to 100%, and the specificity ranged from 64 to100%. The combined sensitivity and specificity estimates of KAtex were 77% (95% CI, 70-83%), and 97% (95% CI, 93-97%), respectively. Comparing the performance of the test by region suggests a significant difference where the lowest and highest sensitivities are reported from Nepal/Tunisia and Europe/Middle East respectively (p < 0.05). On the other hand, the lowest and highest rates of specificity were reported from Sudan and America/Middle East respectively. The overall specificity of KAtex is satisfactory. However, KAtex suffers from low sensitivity and this shortcoming should be improved. The test provides a rapid and simple diagnosis of VL and improvement of its sensitivity deserve further studies.

Production of Monoclonal Antibody Against Excretory-Secretory Antigen of Fasciola hepatica and Evaluation of Its Efficacy in the Diagnosis of Fascioliasis.

Parasitological methods are not helpful for the diagnosis of fascioliasis in acute and invasive periods of the disease. Detection of coproantigens seems to be a suitable alternative approach in the diagnosis of fascioliasis. The present study aimed to develop a reliable antigen detection system, using monoclonal antibodies raised against excretory-secretory (ES) antigen of Fasciola hepatica, for the diagnosis of fascioliasis. Fasciola adult worms were collected from the bile ducts of infected animals. Species of the fluke was determined by polymerase chain reaction-restriction fragment length polymorphism (RFLP-PCR). ES antigen of F. hepatica was prepared. For production of monoclonal antibodies, mice were immunized with ES antigens of F. hepatica. Spleen cells from the immunized mice were fused with NS-1 myeloma cells, using polyethylene glycol. Hybridoma cells secreting specific antibody were expanded and cloned by limiting dilution. Moreover, polyclonal antibody was produced against F. hepatica ES antigen in rabbits. A capture enzyme-linked immunosorbent assay (ELISA) system, using produced monoclonal antibody, was designed and stool samples of infected animals along with control samples were tested by the system. The capture ELISA detected the coproantigen in 27 of 30 (90%) parasitologically confirmed fascioliasis cases, while 4 of 39 (10.25%) samples infected with other parasitic infections showed a positive reaction in this system. No positive reactivity was found with healthy control samples. Accordingly, sensitivity of 90% and specificity of 94.2% were obtained for the capture ELISA system. The results were compared with those obtained with commercial BIO-X ELISA, and a very good (kappa = 0.9) agreement was found between the commercial kit and the developed capture ELISA. Findings of this study showed that the produced monoclonal antibody has appropriate performance for the detection of Fasciola coproantigen in stool samples and can be appropriately used for the diagnosis of fascioliasis.

Seroprevalence of cystic echinococcosis in blood donors in Fars province, southern Iran.

Cystic echinococcosis (CE) is one of the most important zoonotic diseases in different geographical areas of the world including Iran. The current study aimed to assess the seroprevalence of human cystic echinococcosis (CE) in healthy blood donors in Fars province, southern Iran. A total of 1068 serum samples were collected from blood donors from five blood service centers of Fars province in south of Iran. Antigen B was prepared from sheep hydatid cyst fluid and collected sera were evaluated for anti-hydatid cyst antibodies, using antigen-B ELISA. Demographic features of the participants were also recorded during the sample collection. Anti hydatid cyst antibodies were detected in sera of 60 out of 1068 blood donors corresponding to overall seroprevalence of 5.6% in this population. Rate of seroprevalence was 6.7% in females and 5.5% in males. The highest rate of infection (8.3%) was found in age group of higher than 50 years old. There were no significant differences between seropositivity to CE and sex, age or place of residence of the participants (P > 0.05). Findings of this study showed that the rate of CE infection in Fars province, southern Iran, is relatively high. Some of these seropositive cases may have active hydatid cyst in their tissues without overt sign or symptoms.

Helminth Infections of Rodents and Their Zoonotic Importance in Boyer-Ahmad District, Southwestern Iran.

BACKGROUND: Rodents are considered as reservoirs of various zoonotic diseases including helminthic infections. The current study aimed to evaluate the prevalence of helminth infections in rodents, in Boyer-Ahmad district, Southwestern Iran. METHODS: Overall, 52 rodents were captured from various areas of the district by Sherman live traps. The animals were then euthanized and dissected. During necropsy, each organ was examined macroscopically for presence of any cyst or visible parasite. The gastrointestinal tract was removed and their contents were evaluated for larva or adult worms. Trichinella larvae in the rodents' muscles were investigated by both digestion and pathological methods. RESULTS: Twenty-eight (53.8%) of the trapped rodents were male. The rodents were including 25 (48.1%) Meriones persicus, 1(1.9%) Calomyscus bailwardi, 1 (1.9%) Arvicola terresterris, 7 (13.5%) Rattus rattus, 8 (15.4%) R. norvegicus, and 10 (19.2%) Apodemus sylvaticus. Of them, 38 (73.0%) were infected with at least one helminth. Collected rodents were infected with Hymenolepis diminuta (50%), Hymenolepis nana fraterna (28.8%), Skrjabinotaenia sp. (15.4%), Anoplocephalidae sp. (15.4%), Cysticercus fasciolaris (5.8%), Trichuris muris (36.5%), Aspiculuris tetraptera (15.4%), Syphacia sp. (5.7%), Rictularia sp. (15.4%), Trichostrongylus sp. (3.8%), and Gongylonema sp. (3.8%). M. persicus was the most (84%) infected rodent, yet the differences between rodent genus and helminth infectivity were not statistically significant (P>0.05). CONCLUSION: The rodents in Boyer-Ahmad district are infected with different helminths infections that some of them are recognized as threat to human health.

Seroprevalence and Molecular Evaluation of Toxoplasmosis in Patients Undergoing Chemotherapy for Malignancies in the Bushehr Province, Southwest Iran.

BACKGROUND: Toxoplasmosis is a life-threatening infection in organ transplant recipients, people receiving corticosteroid or radiation therapy, people with malignancies, and AIDS patients. OBJECTIVES: The current study aimed to determine the prevalence of toxoplasmosis in patients receiving chemotherapy for malignancies in the Bushehr province of southwest Iran. METHODS: Blood samples were taken from 86 patients who were continuously referred to the chemotherapy center in Bushehr province and evaluated by ELISA to determine anti-Toxoplasma IgG and IgM antibodies. Moreover, a blood buffy coat of each sample was assessed by polymerase chain reaction (PCR), targeting a 529 bp gene of T. gondii. PCR products of the positive samples were sequenced to determine the genotype of the parasite. RESULTS: Anti-Toxoplasma IgG antibodies were detected in the sera of 21 (24.4%) cases. All of the patients were negative for anti-Toxoplasma IgM antibodies. No statistically significant correlation was found between seropositivity to Toxoplasma and duration of chemotherapy or having contact with cats. PCR detected a 529 bp band of T. gondii in the buffy coats of two out of 86 (2.3%) cases. The sequence analysis demonstrated that both cases were 95% identical to type III (VEG strain) of T. gondii. CONCLUSIONS: Findings of this study demonstrated the presence of type III T. gondii in the buffy coats of patients undergoing chemotherapy. Given that toxoplasmosis is a life-threatening infection in immunocompromised patients, these patients should be screened for toxoplasmosis before and during chemotherapy to prevent acute toxoplasmosis.

Visceral Leishmaniasis in Southwestern Iran: A Retrospective Clinico-Hematological Analysis of 380 Consecutive Hospitalized Cases (1999-2014).

Visceral Leishmaniasis (VL) is an endemic parasitic disease and remains as a major health concern in southwestern Iran. The current study describes clinico-hematological, epidemiological and therapeutic features of VL cases, admitted to university-affiliated hospitals, during 1999-2014 in Fars province, southwestern Iran. A total of 380 VL cases were recorded during a 16 years period, giving an average annual admission of 23.75 cases/year in which 217 (57.1%) were male and 163 (42.9%) were female. Mean age of the patients was 3.7 years. The majority of the cases (91.5%) were ≤ 5 years old. Bone-marrow aspiration detected Leishmania amastigotes only in 26.6% of cases. Fever (98.1%), abdominal protrusion (65.1%) and hepatosplenomegaly (63.7%) were the most common clinical presentations of the patients. Pancytopenia was noted in 43.1, anemia in 87.3 and thrombocytopenia in 64% of cases. Increase in the level of AST (aspartate aminotransferase), ALT (alanine aminotransferase), alkaline phosphatase, LDH (lactate dehydrogenase) and CRP (C-Reactive Proteins) were seen in 84.9, 53.6, 44.4, 72.5 and 83.1% of cases, respectively. Mortality was noted in 5.3% of cases. Deranged haemato-biochemical parameters including total and direct bilirubin, PLT (platelet) and pancytopenia were significantly contributed to mortality from VL. Moreover, clinical features such as severe splenomegaly as well as bacterial infections were meaningfully contributed to death from VL. The majority of patients (74.9%) were treated with meglumine antimoniate. Amphotericin B was administrated in 59 of cases, 11 of them were initially treated with meglumine antimoniate with a shift to amphotericin B, because of treatment failure. Findings of the current study demonstrated that VL is present in southwest of Iran with a fairly continual rate during the last 16 years period. Deranged haemato-biochemical parameters along with severe splenomegaly contributed to mortality from VL.

Protozoan Parasites of Rodents and Their Zoonotic Significance in Boyer-Ahmad District, Southwestern Iran.

Backgrounds. Wild rodents are reservoirs of various zoonotic diseases, such as toxoplasmosis, babesiosis, and leishmaniasis. The current study aimed to assess the protozoan infection of rodents in Boyer-Ahmad district, southwestern Iran. Materials and Methods. A total of 52 rodents were collected from different parts of Boyer-Ahmad district, in Kohgiluyeh and Boyer-Ahmad province, using Sherman live traps. Each rodent was anesthetized with ether, according to the ethics of working with animals, and was dissected. Samples were taken from various tissues and stool samples were collected from the contents of the colon and small intestines. Moreover, 2 to 5 mL of blood was taken from each of the rodents and the sera were examined for anti-Leishmania antibodies, by ELISA, or anti-T. gondii antibodies, by modified agglutination test (MAT). DNA was extracted from brain tissue samples of each rodent and PCR was used to identify the DNA of T. gondii. Results. Of the 52 stool samples of rodents studied by parasitological methods, intestinal protozoa infection was seen in 28 cases (53.8%). From 52 rodents, 19 (36.5%) were infected with Trichomonas, 10 (19.2%) with Giardia muris, and 11 (21.2%) with Entamoeba spp. Also, 10 cases (19.2%) were infected with Blastocystis, 3 (5.8%) were infected with Chilomastix, 7 (13.5%) were infected with Endolimax, 1 (1.9%) was infected with Retortamonas, 3 (5.77%) were infected with T. gondii, and 6 (11.54%) were infected with Trypanosoma lewisi. Antibodies to T. gondii were detected in the sera of 5 (9.61%) cases. Results of the molecular study showed T. gondii infection in 3 (5.77%) of the rodents. Findings of this study showed that rodents in Kohgiluyeh and Boyer-Ahmad province, southwestern Iran, are infected with several blood and intestinal parasites; some of them might be potential risks to residents and domestic animals in the region.

Clinical and Molecular Evaluation of a Case of Giant Primary Splenic Hydatid Cyst: A Case Report.

Spleen is an unusual location for hydatid cyst. Here we report a case of primary splenic hydatid cyst in a 41-yr-old Iranian woman from Yasuj, southwest of Iran. The patient had been admitted to Shahid Beheshti Hospital because of abdominal pain. Abdominal sonography revealed a hypoechoic lesion of 150 X 130 mm in the spleen, suggestive of hydatid cyst. Splenectomy was performed for the patient and surgical interventions revealed a hydatid cyst occupying most of splenic parenchyma. She was discharged on the 5 day of her operation. Postoperative diagnosis and confirmation of hydatid cyst was done by histopathological, molecular and serological approaches. Histopathological evaluation revealed the classical laminated layer of hydatid cyst. DNA was extracted from a part of cyst and PCR amplified. Sequencing and analysis of PCR product revealed that the isolate has the most similarity with G1 strain of Echinococcus granulosus. Patient's serum was positive for IgG anti-hydatid cyst antibodies, using antigen-B ELISA.

Severe congenital toxoplasmosis: a case report and strain characterization.

We report a fatal congenital toxoplasmosis case in an Iranian woman in the south of Iran. A pregnant mother had been admitted at the 15th week of her pregnancy on account of a febrile illness, symptoms of common cold, and enlargement of submandibular lymph nodes. Serological testing of the mother's serum revealed positive IgG and IgM anti-Toxoplasma antibodies. Amniotic fluid was taken and evaluated by polymerase chain reaction (PCR) assay with a direct amplification of the Toxoplasma URPT gene which was found to be positive. Sequencing and analysis of PCR product revealed that the isolate has the most similarity with type I of Toxoplasma gondii. Fetal scan showed anomaly in fetus including mild hydrocephaly. Termination of the pregnancy was suggested by the physician and pregnancy was terminated 178 days after conception.

Epidemiology of Human Fascioliasis and Intestinal Helminthes in Rural Areas of Boyer-Ahmad Township, Southwest Iran; A Population Based Study.

BACKGROUND: Fascioliasis, caused by Fasciola hepatica and F. gigantica, is one of the most important plant and water borne disease in Iran. This study aimed to determine the prevalence of fascioliasis and intestinal helminthes in inhabitants of rural areas of Boyer-Ahmad in Kohgiluyeh and Boyer-Ahmad province in Southwest of Iran. METHODS: Stool samples (1025) were collected from inhabitant of 50 randomly selected villages in Boyer-Ahmad Township. Samples were evaluated with modified Telemann and formalin-ethyl acetate methods. Blood samples of Fasciola positive cases were assessed with ELISA and Western blotting. DNA was extracted from Fasciola eggs from stool of positive individuals and evaluated by molecular (PCR) method and the PCR products were sequenced and analyzed. RESULTS: Of the 1025 participants, 473 (46.1%) were male and 552 (53.9%) were female. The mean age of the subjects was 20.25 (±15.86) years. Fasciola eggs were detected in stools of two cases (0.19%). Blood samples were obtained from the Fasciola positive cases and their infection was further confirmed by ELISA and Western blotting. Molecular analysis revealed that both cases are infected with F. hepatica. Furthermore, seven of participants (0.68%) were found to be infected with H. nana, 4 cases (0.39%) with E. vermicularis, and one case (0.09%) with Trichuris trichiura. CONCLUSION: Findings of this study showed that Boyer-Ahmad district is one of the endemic areas of human fascioliasis in Iran. The study also documented that the rate of helminthic infections in rural areas of the district has drastically declined over the past years.

Utility of Western Blot Analysis for the Diagnosis of Cutaneous Leishmaniasis.

BACKGROUND: Cutaneous leishmaniasis (CL) is a parasitic disease with a relatively wide distribution in different areas of the world, including Iran. The parasite is mainly diagnosed microscopically, but serological approaches might be useful for diagnosis as well. This study aimed to assess the efficacy of an immunoblotting system for serodiagnosis of cutaneous leishmaniasis in Iran. METHODS: Sixty-one sera samples from parasitologically confirmed CL patients and 50 sera samples from healthy controls along with 50 sera sample from non-CL patients were collected. Native strain of Leishmania major was cultured in Schneider medium and soluble Leishmania antigens were prepared from amastigotes-like parasites. All of sera samples were evaluated by an immunoblotting system. RESULTS: Components of 14 to 135 kDa were detectable by the sera of CL patients. From 61 sera of CL patients, 59 cases (96.7%) detected a 63 kDa subunit and 51 cases (83.6%) recognized a 32-35 kDa component. Among all subunits, the 63 kDa band showed the highest sensitivity (96.7%) and a 75 kDa band had the highest (98%) specificity. CONCLUSION: Immunoblotting has a satisfactory performance in diagnosis of CL and this test can be used, as an aid, for proper diagnosis of CL.

Performance of an ELISA and indirect immunofluorescence assay in serological diagnosis of zoonotic cutaneous leishmaniasis in iran.

Serological assays have been extensively evaluated for diagnosis of visceral leishmaniasis (VL) and considered as a routine method for diagnosis of VL while these methods are not properly evaluated for diagnosis of cutaneous leishmaniasis (CL). This study aimed to assess the performance of indirect immunofluorescent-antibody test (IFA) and enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of cutaneous leishmaniasis in Iran. Sixty-one sera samples from parasitologically confirmed CL patients and 50 sera from healthy controls along with 50 sera from non-CL patients were collected. Antigen was prepared from promastigotes and amastigotes of Leishmania major. IFA was used to detect anti-Leishmania IgG while ELISA was used to detect anti-Leishmania IgM, total IgG, or IgG subclasses (IgG1 and 4). ELISA, for detection of total IgG and IgM, showed sensitivity of 83.6% and 84.7% and specificity of 62.7% and 54.6%, respectively. Sensitivity and specificity of ELISA for detecting IgG1 and IgG4 were 64%, 75% and 85%, 49%, respectively. Sensitivity and specificity of IFA were 91.6% and 81%. Conclusion. Findings of this study demonstrated that serological test, especially IFA, can be used for proper diagnosis of CL.