Genotyping of Hydatid Cyst Isolated from Human and Domestic Animals in Ilam Province, Western Iran Using PCR-RFLP.

BACKGROUND: Hydatidosis or cystic hydatid disease is one of the most important diseases in human and animals. Identification of strains is important for improvement of control and prevention of disease. The aim of this study was to determine the strains isolated from human and domestic animals in Ilam Province, Iran, using PCR-RFLP method. METHODS: Respectively, 30 and 4 animal and human hydatid cysts were collected from different slaughterhouses and hospitals of the province. Protoscolices were separated and their DNA genome was extracted by extraction kit. rDNA-ITS1 of each isolated samples was duplicated by BD1(Forward) and 4s (Reverse) Primers. PCR products were studied by electrophoresis and then were digested using TaqI, HpaII, RsaI and AluI restriction enzymes. RFLP products were studied using electrophoresis on 1% agar gel. RESULT: A fragment of 1000bp was produced from amplification of rDNA-ITS1 of protoscolices using PCR method. After digestion of PCR product by AluI enzyme, 200bp and 800bp, by RsaI, 655bp and 345bp and by HpaII 700bp and 300bp sizes were obtained. TaqI enzyme had no change in fragment size and it remained 1000bp. Considering the method, Ilam strains was specified as E. granulosus sensu stricto (G1-G3). CONCLUSIONS: Although sheep strain (G1) is dominated in human and different animal in Iran and the world, but more efforts should be done to clarify the true genotype of Ilam strains specified as E. granulosus sensu stricto (G1-G3).

Seroprevalence of human hydatidosis using ELISA method in qom province, central iran.

BACKGROUND: The objective of this study was to determine the prevalence of cystic echinococcosis (CE) in Qom Province, central Iran using ELISA test. METHODS: Overall, 1564 serum samples (800 males and 764 females) were collected from selected subjects by randomized cluster sampling in 2011-2012. Sera were analyzed by ELISA test using AgB. Before sampling, a questionnaire was filled out for each case. Data were analyzed using Chi-square test and multivariate logistic regression for risk factors analysis. RESULTS: Seropositivity was 1.6% (25 cases). Males (2.2%) showed significantly more positivity than females (0.9%) (P= 0.03). There was no significant association between CE seropositivity and age group, occupation, and region. Age group of 30-60 years encompassed the highest rate of positivity. The seropositivity of CE was 2.1% and 1.2% for urban and rural cases respectively. Binary logistic regression showed that males were 2.5 times at higher risk for infection than females. CONCLUSION: Although seroprevalence of CE is relatively low in Qom Province, yet due to the importance of the disease, all preventive measures should be taken into consideration.

Assessing the effect of highly cited papers on the impact factor of journals in the field of public health.

BACKGROUND: The aim of this study was to appraisal the effect of highly cited papers in the field of public health and find out whether the unusual citations affect the ranking order of the journals in this field or not. A total number of 142 journals titles were listed in Journal Citation Report (ISI Thomson) in the field of "Public, Environmental & Occupational Health". All but one of them had published papers at least for a year from 2009 to 2010. Journal title, number of citations and publication year of 45685 papers were collected from ISI web of knowledge database at December 25, 2011. About half of the papers (23226) had no citations and 89.4% (40835) had less than 6 citations. We concluded that the ranking of journals in the field of public health is not affected by the individual papers with unusual number of citations.

The prevalence of honorary and ghost authorships in Iranian bio-medical journals and its associated factors.

BACKGROUND: The purpose of this study was to detect the prevalence of ghost and honorary authors and its determinant factors in bio-medical journals of Iran. METHODS: The study was done in 2009-10 in Tehran, Kerman, and Iran Medical Universities, Iran. We contacted the first or corresponding authors of the papers had published papers in the recent two issues of Iranian Journal of Public Health, Journal of Kerman University of Medical Sciences, and Tehran University Medical Journal. They explored the role of each coauthor and others who had done mouthing for the paper. Then, according to ICMJE criteria, we counted how many of them are real, honorary or ghost author. For the analysis, we utilized two databases. One included articles as the records and the other included authors as the records. RESULTS: From 124 articles, with 536 authors, 301 (56.1%) were honorary authors. Each article had 4.35 authors on average, while 2.4 of them were honorary authors. The percentage of honorary author in basic science articles was about 6% more than the articles of clinical sciences. Moreover, 89% of articles had at least one honorary author. About 20% of all articles had more than three honorary authors. Besides, 25 (21.43%) authors confessed they had colleague(s) omitted from the authors list, while only one (0.81%) of them met the authorship criteria. The percentage of agreement between the corresponding and the remaining authors on the number of honorary of the authors was about 47.4% (Kappa= 0.27, P= 0.01). CONCLUSION: It seems that the present data might assist the authorities to make a decisive decision on amending the process of authorship in Iran.

Molecular Diagnosis of Strongyloides stercoralis Infection by PCR Detection of Specific DNA in Human Stool Samples.

BACKGROUND: Strongyloidiasis is mostly an asymptomatic infection and diagnosis of latent infections is difficult due to limitations of current parasitological and serological methods. This study was conducted to set up a PCR-based method for molecular diagnosis of Strongyloides stercoralis infection by detection of copro-DNA in stool samples. METHODS: A total of 782 fresh stool samples were collected and examined by agar plate culture. Among those sixteen stool samples, which confirmed to be infected with S. stercoralis were examined as positive control to set up each single and nested PCR, using two primer sets designing to amplify partial ribosomal DNA of S. stercoralis genome. Since, single PCR method yielded higher efficacy in detecting positive samples, in the second step, 30 stool samples, which found negative for S. stercoralis by agar plate culture of single stool sample, were examined by single PCR. Data analysis was performed using McNemar's χ(2) test, with consideration of a P-value of <0.05 as indication of significant difference. RESULTS: In amplification of DNA extracted from stool samples, single PCR detected S. stercoralis DNA target in all 16 positive samples, while nested PCR amplified DNA in only 75% of samples. In the second step, single PCR amplified S. stercoralis extracted DNA in 5 out of 30 samples which were negative by coproculture. CONCLUSION: Single PCR method amplifying a short (100bp) target represented more efficacies for detection of S. stercoralis in faecal examination compared to agar plate culture and nested PCR, which amplified longer target.

Establishment of Hymenolepis diminuta Life Cycle to Provide Parasite Mass Production.

BACKGROUND: The main object of this experimental work was to practise laboratory production both adult and the larval stage of Hymenolepis diminuta with conventional modification to make further studies easier. MATERIALS #ENTITYSTARTX00026; METHOD: Adults H. diminuta were collected from urban rats in Tehran, Iran. The beetles became infected using blended gravid segments with flour as bait. Cysticercoids have been saved after precise dissection of invertebrate hosts. The exposure of infected beetles to laboratory rats was performed to establish the life cycle. RESULT: Out of 57 collected rats, three rats were infected with H. diminuta. Almost all exposed beetles found infected with the larval stage of parasite. About one-month later H. diminuta eggs were seen in stool examination of laboratory rats. CONCLUSION: Rare human occurrence of H. diminuta along with light level of clinical manifestation of this parasite, underestimate the concerns toward its public health importance. Nowadays, various field of studies, such as biochemistry with special focuses on the capability of H. diminuta tegument absorption have performed apart from parasitological views alone. In the present study, establishment of this parasite life cycle has practically provided the access of adult and cysticercoid stages of the tapeworm in further researches.

Seroepidemiological study of human hydatidosis in meshkinshahr district, ardabil province, iran.

BACKGROUND: The aim of this study was to conduct a sero-epidemiological survey in Meshkinshahr, Ardabil Province, northwestern Iran to detect the rate of hydatidosis in the city and nearby villages. Literature shows that no such study has been conducted so far. METHODS: Overall, 670 serum samples were collected from 194 males and 476 females from patients referred to different health centers of the region. All patients filled out a questionnaire and an informed consent. Sera were analyzed using indirect-ELISA test. Ten µg /ml antigens (Antigen B derived from hydatid cyst fluid), serum dilutions of 1:500 and conjugate anti-human coombs with 1:10000 dilutions were utilized to perform the test. Data analysis was conducted using SPSS software ver. 11.5. RESULTS: The seroprevalence of human hydatidosis was 1.79% by ELISA test in the region. This rate for females was 1.68% and males 2.6%, respectively. There was no significant difference as regards all factors studied and the seropositivity. According to job, farmers and ranchmen had the highest rate of infection as 3.17%. The sero-prevalence of infection was 2.6%% in illiterate people which showed the highest rate. As regards residency, urban life showed no significant difference with rural life (1.1% vs. 2.58%). Age group of 69-90 yr old, with 4.62% as prevalence had the highest rate of positivity. CONCLUSION: Obtained sero-prevalence of hydatidosis shows more or less a resemblance to other cities of Iran, although due to the specific condition of the city we expected more rate of sero-positivity.

Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2).

BACKGROUND: In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species. METHODS: The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety trematodes of Fasciola were collected during abattoir inspection from livers of naturally infected sheep and cattle from Khorasan, East Azerbaijan, and Fars provinces in Iran. After DNA extraction, PCR was performed to amplify region ITS1, 5.8S rDNA, ITS2. To select a suitable restriction enzyme, we sequenced and analyzed the PCR products of F. hepatica and F. gigantica samples from sheep and cattle. Tsp509I fast digest restriction enzyme was selected for RFLP method that caused the separation specifically of Fasciola species. RESULTS: The fragment approximately 1000bp in all of the Fasciola samples was amplified and then digested with the Tsp509I restriction endonuclease. Seventy F. hepatica and 20 F. gigantica were identified of total 90 Fasciola isolates. CONCLUSION: The new PCR-RFLP assay using Tsp509I restriction enzyme provides a simple, practical, fast, low cost, and reliable method for identification and differentiation of Fasciola isolates.

Heavy Worm Burden of Moniliformis moniliformis in Urban Rats with Histopathological Description.

BACKGROUND: Due to scarcity of human reports, we took advantage of the heaviest infection of M. moniliformis in rats, to describe histopathological and microanatomical valuable useful keys while confronting human occurrences. METHODS: Samples were obtained from captured rats in Tehran, capital of Iran, during two decades. Tissue sections were performed through hematoxylin and eosin staining to describe histopathological changes in rat's intestines. RESULTS: Totally, nine rats were found infected with M. moniliformis amongst 272 obtained rats. Heavy infection has been distinguished in 2 individuals with parasite burden of 141and 73 adult worms. Cross sections of worms within the lumen show mucosal thickness, infiltration of eosinophilic leukocyte and increase in goblet cells. CONCLUSION: Beyond the uncommonness of human infection with M. moniliformis unitended infections should not be ignored. Abundance of rats and roaches as definite and intermediate hosts must be considered particularly in countries with poor hygiene.

Evaluation of the stability of coated plates with antigen at different temperatures and times by ELISA test to diagnose fasciolosis.

BACKGROUND: Considering that ELISA method presently is the test of choice for diagnosis of fasciolosis, the present study was undertaken to evaluate the maximum validity of coated plates at different temperatures and different times during one year of evaluation. METHODS: Serum samples of patients infected with fasciolosis (n=10), hydatidosis (n=5), toxocariasis (n=5), and negative control sera (n=5) were examined. Two series of plates were considered. The first series were coated with Fasciola homogenate Ag 12 ug/ml, and after some steps were blocked with gelatin and preserved at different temperatures as -80°C, -20°C, -4°C and +4°C. The 2(nd) series were treated under the same criteria but were not blocked with gelatin. Each series were examined by ELISA test from 1(st) month to 12(th) month. Sera with 1:125 dilution, and peroxidase-conjugated goat anti-human IgG diluted 1:10000 were considered optimum. RESULTS: To ease reporting the results and due to many similarities only results related to 1(st), 6(th) and 12(th) months were analyzed and sensitivity, specificity plus cut-off were determined for each series separately. CONCLUSION: Preserving the coated plates, while unblocked at -80°C for 6-8 months is pertinent and functional and in that case, we can be sure the best out put would be applicable.

Serological Evaluation of EgAgB16 kDa, a Recombinant Antigen from Echinococcus granulosus for Diagnosis of Human Hydatidosis.

BACKGROUND: Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16 kDa) from Echinococcus granulosus (Iranian G1 strain) and its evaluation by ELISA test. METHODS: DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain) protoscoleces collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected individuals (n=72), healthy individual (n=48), toxoplasmosis (n=4), strongyloidosis (n=4), kala-azar (n=5) and tuberculosis (n=5) were examined using this recombinant antigen. RESULTS: Recombinant protein was purified by affinity chromatography using His-Tag column. After purification, recombinant protein was confirmed by western blot analysis using His Tag monoclonal antibody or hydatid positive human serum. The sensitivity, specificity; positive and negative predictive values were calculated as 93.5%, 95.6%, 96% and 92.9%, in that order. The cut-off point was detected 0.3 for rAgB16. CONCLUSION: While the produced recombinant AgB16 kDa showed promising results in diagnosing human hydatidosis, but more investigations should be implemented to reach an accurate gold standard.

Ocular dirofilariasis, a case report.

Accidental infection with animal filarial worms in humans is a dilemma for clinicians and parasitologists throughout the world. To date a variety of such rare parasitoses have been reported mostly in tropics and subtropics. Human dirofilariasis is among those unusual zoonotic infections that occasionally have been observed in the eye and in subcutaneous areas exhibiting with nodule formation. Filarial worms are transmitted to humans through invertebrate biological vectors such as certain species of mosquitoes. The present report describes a peculiar case of ocular dirofilariasis in a 49-year-old man resident in Iran.

Neglected human fascioliasis case in a visceral leishmaniasis endemic area, north-Western iran.

Ardabil Province, northwestern Iran, is endemic for visceral leishmaniasis of Mediterranean type from many years ago. This situation might cause to underestimate other important diseases, which occur sporadically within the same area. In this case, a 6-year-old male patient in this area was treated as infected with visceral leishmaniasis initially with seropositive result for visceral leishmaniasis. Afterwards, histopathological study of crashed material and tissue sections surprisingly revealed numerous liver fasciolid fluke eggs. This case demonstrates that in an area endemic for a given parasitoses, other important infectious diseases must be considered as well.