Prevalence and genetic characterization of Cryptosporidium spp. In diarrheic children from Gonbad Kavoos city, Iran

Background: Cryptosporidium is an intestinal protozean parasite causing water-borne and foodborne outbreaks of diarrheal diseases. The present study was per-formed in order to find prevalence and subtypes of Cryptosporidium among children with diarrhea in Gonbad Kavoos City, Northern Iran. Methods: Diarrheic samples were collected from 547 children. The initial parasi-tological diagnosis was made based on detection of oocysts using the modified Ziehl-Neelsen acid-fast staining method. The positive microscopically samples were selected for sequence analysis of partial 60 kDa glycoprotein (gp60) gene. Results: Out of 547 collected samples, 27 (4.94) were positive for Cryptosporid-ium oocysts. Fifteen from 27 positive samples successfully amplified in PCR. Se-quences analysis of gp60 gene in 15 Cryptosporidium isolates revealed that all of them (100) were C. parvum. The results showed three subtypes of IIa subtype family (7 cases) including IIaA16G2R1, IIaA17G1R1, IIaA22G3R1 and one subtype of IId subtype family (8 cases). The most common allele was IId A17G1d (53.3). Conclusion: The predominance of zoonotic subtype families of C. parvum species (IIa, IId) in the present study is in concordance with previous studies in Iran and emphasizes the significance of zoonotic transmission of cryptosporidiosis in the country. © 2015, Tehran University of Medical Sciences (TUMS). All rights Reserved

An improved microculture method for diagnosis of cutaneous leishmaniasis

This study evaluated the performance of three diagnostic methods for cutaneous leishmaniasis (CL). Patients who came to the Health Center Laboratory of Gonbad-e-Qabus in Golestan Province, Iran, were enrolled in the study. Skin scraping smear, improved microculture (IMC) and polymerase chain reaction (PCR) were performed. A total of 303 subjects were recruited, among whom 273 subjects fulfilled the criteria for CL. Sensitivity and specificity were 88.8 % (95 % CI = 84.2–92.2 %) and 100.0 % for smears, 98.4 % (95 % CI = 96.1–99.1 %) and 100.0 % for IMC, both of them 100.0 % for PCR. Although, PCR was relatively more sensitive than the IMC, the high correlation (agreement = 96 %, Kappa = 0.82) between IMC and PCR along with the advantages of simplicity, rapidity, adequate sensitivity and as a needle free method, offers the IMC as a valuable alternative diagnostic method for PCR in diagnosis of CL. © Indian Society for Parasitology 2013

Identification of Leishmania species isolated from human cutaneous Leishmaniasis in Gonbad-e-Qabus city using a PCR method during 2006-2007

"n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 st1":*{behavior:url(#ieooui) } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Cutaneous Leishmaniasis is endemic in plenty of Iranian provinces. This study aimed to determine the epidemiological status of the cutaneous Leishmaniasis outbreak, isolation and identification of the parasite using a PCR method in burden rural areas of Gonbad-e-Qabus County, north Iran. "n"nMethods: Data was collected on the prevalence of scars and ulcers over a period of three months among 6990 inhabitants of five villages around Gonbad-e-Qabus county, north Iran, during 2006-2007. Cultured promastigotes were identified using PCR technique. ITS1 and ITS2 of Non Coding Transcribed region at ribosomal DNA of 46 Leishmania isolates were amplified and the PCR products were separated by electrophoresis in 1.5% agarose gel (200 mA, 140 V), visualized by staining with ethidium bromide, and photographed. To confirm the PCR findings, six Leishmanis isolates were injected individually into two BALB/c mice."n"nResults: Among 6990 inhabitants of the five villages, 62.9% had scars and 0.5% had active lesions. The most highly infected age group was 0-10 years and nobody was infected in individuals more than fifty years of age. Individuals 11 to 20 years of age were the most highly infected age group. The results showed that from 46 isolates, all (100%) were L. major in comparison to reference strains and all of them could produce ulcer at the base tail of BALB/c mice, 4-12 weeks after inoculation."n"nConclusions: According to this study, cutaneous Leishmaniasis due to Leishmania major is endemic in Gonbad-e-Qabus county, north Iran. The results were confirmed by active lesions induced in BALB/c mice

Identification of Leishmania species isolated from human Cutaneous leishmaniasis in gonbad-e-Qabus city using a PCR method during 2006-2007

Background: Cutaneous Leishmaniasis is endemic in plenty of Iranian provinces. This study aimed to determine the epidemiological status of the cutaneous Leishmaniasis outbreak, isolation and identification of the parasite using a PCR method in burden rural areas of Gonbad-e-Qabus County, north Iran. Methods: Data was collected on the prevalence of scars and ulcers over a period of three months among 6990 inhabitants of five villages around Gonbad-e-Qabus county, north Iran, during 2006-2007. Cultured promastigotes were identified using PCR technique. ITS1 and ITS2 of Non Coding Transcribed region at ribosomal DNA of 46 Leishmania isolates were amplified and the PCR products were separated by electrophoresis in 1.5% agarose gel (200 mA, 140 V), visualized by staining with ethidium bromide, and photographed. To confirm the PCR findings, six Leishmanis isolates were injected individually into two BALB/c mice. Results: Among 6990 inhabitants of the five villages, 62.9% had scars and 0.5% had active lesions. The most highly infected age group was 0-10 years and nobody was infected in individuals more than fifty years of age. Individuals 11 to 20 years of age were the most highly infected age group. The results showed that from 46 isolates, all (100%) were L. major in comparison to reference strains and all of them could produce ulcer at the base tail of BALB/c mice, 4-12 weeks after inoculation. Conclusions: According to this study, cutaneous Leishmaniasis due to Leishmania major is endemic in Gonbad-e-Qabus county, north Iran. The results were confirmed by active lesions induced in BALB/c mice

Molecular identification of Leishmania species isolated from patients with cutaneous leishmaniosis in gonbad Kavoos, northeastern of Iran using hSP70 and ITS-based PCR-RFlP

Cutaneous leishmaniosis (CL) is mainly caused by Leishmania major (rural-type) and Leishmania tropica (urban-type). CL is a major health problem in many regions of the world, and it is associated with health complications and economic loss. The identification and differentiation of Leishmania species are critical because the prevention and control methods, as well as management and therapeutic strategies, are different for each type of CL. The present study aimed to identify the parasite species responsible for CL in the study area using ITS1 and HSP70- based PCR-RFLP methods. A total of 147 stained slides were prepared from samples collected from CL patients, and these slides were positive for amastigotes of Leishmania species on microscopic examination. Forty-three Giemsastained slides with 2+ to 4+ grades were selected for molecular studies for the identification of the Leishmania species. DNA was extracted from the selected slides for the molecular studies. The amplification of HSP70 and ITS1 genes was performed by the PCR method. The PCR products were digested with the HaeIII restriction enzyme, and banding patterns of all samples were compared with reference strains. Overall, patterns of all the samples were found to correspond to the reference strains of L. major based on RFLP-PCR targeting HSP70 and ITS1 genes of the parasite, demonstrating the dominance of L. major as the causative agent of zoonotic cutaneous leishmaniosis (zCL) in the study area. This area is endemic for zoonotic CL, and further studies are required to determine the reservoir and natural infection of sand flies in this county

Prevalence of Entamoeba histolytica and Entamoeba dispar in Gonabad City, 2006, Iran

Background: Differential diagnosis of two protozoan parasites Entamoeba histolytica and E. dispar is of great clinical and epide¬miologi¬cal importance, but except in the case of haematophagous trophozoites in acute dysentery, it is not possible to differ¬entiate them by microscopy. The present study was carried out from February 2005 to July 2006 in order to determine the preva¬lence of E. histolytica and E. dispar in Gonbad City, by using a PCR method. Methods: Five hundred and sixty four fecal samples were collected from primary health care centers of Gonbad both urban and rural ar¬eas. The stool specimens were examined by light microscopy (Direct slide smear, Iodine wet mount, Formal-ether concentra¬tion and Trichrome staining) to distinguish E. histolytica/E. dispar complex and differentiate them from other non-patho¬genic intestinal amoebae. Results: The microscopy results of stool exams showed a frequency rate of 23 positive samples (4.07%) for cyst of E. histo¬lytica/E. dis¬par complex. All the microscopy positive isolates appeared to be infected with cyst of E. histolytica/E. dispar com¬plex were cultivated and maintained successfully in HSr + s medium for DNA extraction and identification by the PCR method. The PCR study showed that 16 isolates (69.56 %) of the Entamoeba samples were E. dispar while 7 samples (30.43%) of those microscopy positive samples were not amplified and none of them showed E. histolytica pattern. Conclusion: High frequency rate of E. dispar in this study were in high agreement with the estimation rate of these entamoe¬bas worldwide

Fauna and larval habitats characteristics of mosquitoes (diptera: Culicidae) in Golestan Province, Northeast of Iran, 2014-2015

Background: Mosquitoes (Diptera: Culicidae) is one of the most medically important families of Diptera. The aims of this study were to determine fauna and larval habitat characteristics of mosquitoes in Golestan Province, during 2014-15. Methods: This study was conducted in larval habitats of mosquitoes and installed ovitraps in 14 districts of Golestan Province, Northern Iran in 2015. Samples were collected with a scoop by ladle handling for entomology. The collected larvae were transferred to Laboratory of Medical Entomology in lactophenol solution. Then microscopic slides were prepared using de Faure's formula. Species of each sample was recognized using diagnostic criteria to identify the Culicidae species. Characteristics of larval breeding places were studied based on the habitat type (Permanent or temporary), water conditions (Clear or turbid, stagnant or running), vegetation (out, in, underwater vegetation or without vegetation), sunlight exposure (Full or partial sunlight) and so on. Data were analyzed using SPSS. Results: Overall, 5661 third-and fourth-instars larvae of mosquitoes were collected and 5 genera and 14 species were identified: Anopheles hyrcanus, An. maculipennis, An. pseudopictus, An. superpictus, Culex hortensis, Cx. mimiticus, Cx. perexiguus, Cx. pipiens, Cx. pusillus, Cx. theileri, Cx. tritaeniohynchus, Culiseta longiareolata, Ochlerotatus caspius, Uranotaenia unguiculata. Culex pipiens was recognized as predominant species of the family. Among the detected species, Cx. pusillus reported for the first time from Golestan Province. Conclusion: Due to the high species diversity of Culicidae, ecology of medical important species such as Cx. pipiens and Cx. tritaeniorhynchus needs more investigations. © 2018 Tehran University of Medical Sciences. All rights reserved

A comparative evaluation of regulatory T cells profile among acute and chronic cutaneous leishmaniasis using flow cytometry

Background: Cutaneous leishmaniasis (CL) is described as a major health problem in many countries of the world. Regulatory T cells (Tregs) are characterized as one of immunologic indexes. One of the best methods to determine of Tregs percentage is flow cytometry. The aim of this study was determination of the role of Tregs profile among acute and chronic forms of human CL using flow cytometry analysis. Methods: This study was conducted on 24 patients referred to Laboratory of Leishmaniasis, Tehran University of Medical Sciences, Tehran, Iran with acute and 14 patients with chronic phases of CL as well as 15 healthy individuals as control group in 2015-2016. After microscopic examination, 2 ml of peripheral blood samples were collected for determining percentage of CD4 + CD25+ CD127 low Tregs by using flow cytometry method. Results: Using flow cytometry analysis, the average percentage of Tregs were calculated 5.73, 6.71 and 6.61 for acute, chronic and healthy individuals, respectively. With SPSS software and Scheffe multiple comparison tests, the differences within in these groups are statistically significant (P=0.04) and between the acute and chronic group, there was marginally significant with approximately 91 of confidence level (P=0.088). Conclusion: Marginally differences were found significantly among averages of Regulatory T cells, acute and chronic phases of CL. Further comprehensive studies can be needed to verify the role of Tregs in both phases of CL cases. © 2019, Tehran University of Medical Sciences (TUMS). All rights reserved