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The idea of the existence of Phlebotomus (Larroussius) neglectus (Diptera: Psychodidae) Tonnoir, 1921 in Iran and the skepticism about the existence of Phlebotomus major s.str. Annandale, 1910 had been grown recently in the country. This study reports a combined analysis of mitochondrial and ribosomal DNA target regions of P. major s.l.Annandale, 1910, specimens collected from different parts of Iran. Two different morphotypes were found among the collected samples based on the shape of the aedeagus, ventrally located hairs of the coxite, and parameral sheets. One morphotype seemed similar to P. neglectus Tonnoir 1921 or P. major krimensis Perfiliv1966 (called here MI.N.K.); the other one was similar to P. neglectus and to some extent to P. notus Artemiev & Neronov 1984 (here called MII.N.NO). Cytochrome B, elongation factor 1-alpha, and internal transcribed spacer II loci were amplified, sequenced, and characterized. High sequence homology (98–100%) was observed between P. neglectus and these morphotypes, and phylogenetic analysis was also concordant. Phlebotomus neglectus sequences available in GenBank are located as the sister group of sequences here, particularly near to morphotype MII.N.NO. Moreover, ITS2 locus provides the maximum resolution for differentiation of two morphotypes. Based on achieving results, although a strong support for the presence of P. neglectus was provided, but it is too early to say that P. major s.str. does/does not exist in Iran. This question could be resolved by studying more samples and, most importantly, by comparing the topotypes of P. neglectus and P. major s. str. if possible in the future.

TheExosomes were extracted from ST1-3To our knowledge, this is the first report of the release of exosome-like vesicles by the human parasite

The current study investigated the prevalence and genotypes ofA total of 950 stool samples were examined using the standard formalin-ether concentration technique. All specimens were cultured in Robinson xenic medium. Subsequently, DNA extraction and PCR amplification of subtype specific sequence-tagged site (STS) were conducted.Microscopic examination showed that 86 out of 950 samples (9.05%) were infected withBased on the current findings, ST3 was the most frequent subtype among all positive samples. Having a better understanding of

Rodents and stray cats are the sources of many parasitic infections including T. gondii, for other animals and human. Toxoplasmosis has a wide range of laboratory factors in its intermediate and definite hosts. Regarding the importance of rodents and stray cats as the hosts that spread the Toxoplasma gondii, it is necessary to obtain comprehensive information about these animals in the life cycle of T. gondii. The objective was to investigate the new prevalence of toxoplasmosis among target animals in Iran, using GRA6 gene in combinacion with ELISA avidity. In this study, 286 rodents and 210 stray cats were collected and their heart tissues extracted to obtain DNA, blood samples and IgG Ab of T.gondii parasite. We detected the positive tissue samples in our study by the nested-PCR method. Then, we examined T. gondii IgG ELISA avidity for assessment of toxoplasmosis among rodents and stray cats. This study, was conducted in January to March 2017, based on the prevalence study. The findings revealed that 246/286 (86.01%) of rodents and 180/210 (85.71%) of stray cats were positive by IgG ELISA avidity methods. moreover, 68 rodents samples and 38 stray cats samples were positive concerning the GRA6 Toxoplasma gene; and these positive samples were at intermediate levels for IgG avidity. We concluded that the new prevalence of toxoplasmosis among rodents and stray cats was at high levels, using the serologic method in Northeast of Iran and the results of quantitative ELISA avidity were as the same as those of the nested-PCR for detecting recent toxoplasmosis in these hosts.

Background: Leishmania is a mandatory intracellular pathogen and causing neglected disease. Hence, protection against leishmaniasis by a development vaccine is an important subject. This study aimed to design a poly-epitope vaccine for cutaneous leishmaniasis. Methods: The present study was conducted in the Parasitology Department of Tarbiat Modares University, Tehran, Iran during 2017-2019. Several bioinformatics methods at online servers were used for prediction of different aspects of polyepitope, including, physico-chemical attributes, allergenicity, antigenicity, secondary and tertiary structures, B-cell, T-cell and MHC (I, II) potential epitopes of LACK, LEIF, GP63 and SMT antigens of L. major. Results: After designing the construct (GLSL), the outputs of PTM sites demonstrated that the poly-epitope had 57 potential sites for phosphorylation. Furthermore, the secondary of GLSL structure includes 59.42%, 20.94% and 19.63% for random coil, extended strand and alpha-helix, respectively. The GLSL is an immunogenic protein with an acceptable antigenicity (0.8410) and non-allergen. Afterward, 20 potential epitopes of LACK, LEIF, GP63 and SMT antigens were linked by a flexible linker (SAPGTP), then was synthesized, and sub-cloned in pLEXY– neo2. The results were confirmed the expression of 38.7 kDa poly-epitope in secretory and cytosolic sites, separately. Conclusion: A good expression in the L. tarentulae and confirmation of the GLSL poly-epitope could be a basis for developing a vaccine candidate against leishmaniasis that should be confirmed via experimental tests in BALB/c mice.

Background Plasmodium vivax resistance to chloroquine (CQ) has been reported from many endemic regions in the world. Plasmodium vivax is responsible for 95% of malaria cases in Afghanistan and CQ is the first-line treatment given for vivax malaria. The pvmdr-1 and pvcrt-o (K10 insertion) genes are possible markers for CQ-resistance in P. vivax isolates. There have been no studies done on the presence or absence of molecular markers for CQ-resistance P. vivax in Afghanistan. The present work aimed to evaluate the frequency of mutations in the pvmdr-1 and K10 insertion in the pvcrt-o genes of P. vivax. Methods Plasmodium vivax isolates were collected from Laghman, Baghlan and Khost provinces. For investigation of polymorphisms of desired regions in pvmdr-1 and pvcrt-o genes, sequencing was applied on the PCR products. A new asymmetric qPCR and melting analysis assay based on unlabelled probe developed for scanning of K10 insertion in pvcrt-o gene. Results The analysis of sequencing data of the pvmdr-1 gene showed wild type Y976 and K997 and mutant M958 and L1076 in 33 isolates from three provinces. Of the 36 samples evaluated for K10 insertion in pvcrt-o, 2/18(11%), 0/10(0%) and 0/8(0%) isolates from Laghman, Baghlan and Khost province, respectively, possessed K10 insertion, confirmed by either sequencing and unlabelled probes. Conclusion Two samples with K10 insertion and 33 samples with pvmdr1 polymorphism, indicating on the possibility of CQ resistance in P. vivax populations in Afghanistan. Furthermore, unlabelled probes are simple and inexpensive alternative tools for screening of P. vivax mutations.

Background: Chloroquine (CQ) resistance Plasmodium vivax isolates have been reported from many endemic regions in the world. P. vivax has been reported to be about 95% of the whole malaria in Afghanistan and CQ is prescribing in the first-line treatment of vivax malaria. The pvmdr-1 and pvcrt-o (K10 insertion) genes are the possible markers of CQ-resistance P. vivax isolates. There have been no studies done on the prevalence of molecular markers of CQ-resistance P. vivax in Afghanistan. In the present work, we aimed to evaluate the prevalence of mutations in the pvmdr-1 and K10 insertion in the pvcrt-o genes of P. vivax.Methods: P. vivax isolates were collected from Laghman, Baghlan and Khost provinces. For investigation of polymorphisms of desired regions in pvmdr-1 and pvcrt-o genes, sequencing was applied on the PCR products. We developed a new asymmetric qPCR and melting analysis assay based on unlabeled probe for scanning of K10 insertion in pvcrt-o gene.Results: The analysis of sequencing data of the pvmdr-1 gene showed wild type Y976 and K997 and mutant M958 and L1076 in 33 isolates from three provinces and submitted in GenBank (accession number MK419882-MK419914)Of 36 samples that evaluated for K10 insertion in pvcrt-o, 2/18(11%), 0/10(0%) and 0/8(0%) isolates from Laghman, Baghlan and Khost province possessed K10 insertion, respectively, that confirmed by either sequencing and unlabeled probes and submitted in GenBank (accession number MK292011-MK292046).Conclusion: The existence of 2 samples with K10 insertion and 33 samples with pvmdr1 polymorphism indicating on CQ resistance P. vivax populations in Afghanistan. This can lead to spreading of resistant strains in the society. Furthermore, unlabeled probes are simple and inexpensive alternative tools for screening of P. vivax mutations.

Background: Malaria is threatening more than half of Afghanistan population. Asymptomatic malaria is notable problem against malaria controlling strategies. In this study we evaluated the asymptomatic malaria status in Nangarhar Province, Afghanistan in 2017. Methods: Overall, 296 finger blood samples were taken on DNA Banking Cards and microscopic slides from asymptomatic individuals in Jalalabad city. We used a novel post real time PCR high resolution melting analysis beside microscopy and semi-nested multiplex PCR to evaluate status of asymptomatic malaria in this city. Results: The prevalence of asymptomatic malaria in Jalalabad city was determined 1.7% (5/296), 7.43% (22/296) and 7.78% (26/296) by microscopy, Seminested multiplex PCR and qRT-PCR-HRM, respectively. Out of 26 positive cases were detected by qRT-PCR-HRM, 21, 1 and 4 cases were detected P. falciparum, P. vivax and mixed infection of P. falciparum and P. vivax, respectively. Conclusion: Our data indicating on existence of significant number of asymptomatic reservoirs that assists in prolonged endemicity of the disease. On the other hand, the molecular methods are better alternatives for microscopy especially for monitoring of asymptomatic cases of malaria.

Treatment for toxoplasmosis is not completely successful because of their unwanted side effects, and new treatments are needed. Imiquimod has ability to moderate immune response and used to treat a wide variety of infections and tumors. The aim of the present study was to evaluate the effect of imiquimod on the tachyzoites of T. gondii and infected macrophages in vitro and in BALB/c mice. The viability of T. gondii was assessed in the presence of various concentrations of imiquimod by direct counting after 6 and 24 h. The MTT assay was used to identify the viability of uninfected macrophages. The apoptotic effects were determined with flow cytometry on the tachyzoites and infected macrophages. For evaluation of parasite load in pre-treatment or post-treatment of macrophages Quantitative real time PCR (qPCR) was performed. For in vivo experiments, BALB/c mice received imiquimod before and after challenge with parasites. The mortality rate of mice, parasite numbers in spleen, and the INF-γ and IL-4 cytokine levels in spleen lymphocytes were evaluated. Imiquimod demonstrated anti-Toxoplasma effects by reducing the number of tachyzoites. The results of flow cytometry for drug-treated tachyzoites showed that apoptosis did not rise significantly relative to the control group (p < 0.05). Moreover, apoptosis was enhanced in infected macrophages as the concentration of imiquimod was reduced. The parasitic burden in imiquimod pretreated macrophages was significantly lower than those treated after infection (p < 0.01). A marked reduction was observed in survival rate, parasite load and INF-γ level in BALB/c mice that received imiquimod before parasitic challenge relative to those received drug after parasitic challenge (p < 0.01). Overall, imiquimod in the pretreated group had greater anti-Toxoplasma effects than imiquimod in posttreated group in vitro and in vivo. imiquimod may be considered as a candidate for use against Toxoplasmosis both therapeutically and prophylactically.

Microsporidia are eukaryotic, intracellular obligate parasites that widely involve many organisms including insects, fish, birds, and mammals. One of the genera of Microsporidia is Encephalitozoon, which contains several opportunistic pathogens. Since Encephalitozoon spp. are zoonotic and opportunistic pathogens, it is important to find their reservoir hosts; hence, the current study aimed at isolating and identifying Encephalitozoon spp. in the crows by the light microscopy observations and molecular methods. For this purpose, 36 samples were collected by the dropping method; however, due to the low volume of samples, the total samples were collected in a sterile stool container and the polymerase chain reaction (PCR) was performed to detect Encephalitozoon spp. Accordingly, 300-bp bands, specific to Encephalitozoon spp., were observed and by sequencing E. intestinalis was identified. Crows can be considered as the hosts of E. intestinalis.

Rodents and stray cats are the sources of many parasitic infections including T. gondii, for other animals and human. Toxoplasmosis has a wide range of laboratory factors in its intermediate and definite hosts. Regarding the importance of rodents and stray cats as the hosts that spread the Toxoplasma gondii, it is necessary to obtain comprehensive information about these animals in the life cycle of T. gondii. The objective was to investigate the new prevalence of toxoplasmosis among target animals in Iran, using GRA6 gene in combinacion with ELISA avidity. In this study, 286 rodents and 210 stray cats were collected and their heart tissues extracted to obtain DNA, blood samples and IgG Ab of T.gondii parasite. We detected the positive tissue samples in our study by the nested-PCR method. Then, we examined T. gondii IgG ELISA avidity for assessment of toxoplasmosis among rodents and stray cats. This study, was conducted in January to March 2017, based on the prevalence study. The findings revealed that 246/286 (86.01%) of rodents and 180/210 (85.71%) of stray cats were positive by IgG ELISA avidity methods. moreover, 68 rodents samples and 38 stray cats samples were positive concerning the GRA6 Toxoplasma gene; and these positive samples were at intermediate levels for IgG avidity. We concluded that the new prevalence of toxoplasmosis among rodents and stray cats was at high levels, using the serologic method in Northeast of Iran and the results of quantitative ELISA avidity were as the same as those of the nested-PCR for detecting recent toxoplasmosis in these hosts.

Background: Trichomoniasis is the most common nonviral sexually transmitted human disease that is caused by protozoan Trichomonas vaginalis. Metronidazole is the selective drug in trichomoniasis treatment. However, the reported cases show an increasing trend of drug resistance. This study aimed to evaluate the effect of mango and blueberry extracts on T. vaginalis. Materials and Methods: T. vaginalis was cultured axenically in TYM (Trypticase Yeast Extract) medium supplemented with 10% bovine serum. The effect of mango and blueberry extracts at 50, 100, 200, 400, 800 and 1000 μg.mL -1 on T. vaginalis was studied after 24 and 48 hours. The final numbers of parasite with a hemocytometer and Trypan blue were recorded. Then the value of IC 50 [Half maximal inhibitory concentration] and the lethal percent were calculated. In the present study, the metronidazole was used as positive control. The IC50 value of metronidazole and tinidazole were calculated in the concentrations of 0.02, 0.04, 0.08, 0.16 and 0.32 μg.mL -1 . Results: The final results confirmed the significant effect of all mango and blueberry extracts concentrations on the reduction of parasite numbers (P-value

Cryptosporidium is a world widely distributed parasite which comparatively has a high prevalence in developing countries. The zoonotic potential of some Cryptosporidium species has made the cryptosporidiosis a significant concern to physicians and veterinarians. The occurrence and zoonotic potential of Cryptosporidium species in probable reservoir hosts for man infections was determined by examining faeces of symptomatic and asymptomatic animals. The aim of this study is to screen the presence of Cryptosporidium in fecal sample of free ranging animals in Tehran using Ziehl–Neelsen staining method. The findings indicate that Cryptosporidium are present in 9/50 (18 %) stray cat (Felis catus), 12/50 (24 %) hooded crows (Corvus cornix), 23/180 (12.7 %) rat (Rattus norvegicus and R. rattus) and 1/40 (2.5 %) pigeons (Columba livia). This investigation confirms the potential role of rats, cats, crows and pigeons for zoonotic transmission of human cryptosporidiosis and they must be considered as reservoir hosts which can endanger public health.

Infection with Trichomonas vaginalis is one of the most common sexually transmitted diseases (STDs) in humans. The prevalence of infection in Iran has been reported 0.009–8 % depending on deferent socio-cultural conditions. This study aimed to determine the frequency of T. vaginalis according to age in patients referred to clinics, hospitals and medical diagnostic laboratories in Karaj city, Iran. For this purpose, fifty positive samples were collected from July 2012 to June 2014 from clinics, medical diagnostic laboratories and hospitals, then transferred to laboratory of parasitology and cultured in TYM medium. The results showed that all isolates were successfully cultured. Among 50 positive specimens, 43 cases were female and 7 cases male. The most positive cases (34 %) belonged to the ages over 50 year’s group. The lowest positive cases (2 %) belonged to the ages of less than 20 years group. In Conclusion, unlike other STDs, which have a higher prevalence among adolescents and young adults, the rates of trichomoniasis are more evenly distributed among sexually active women of all age groups. However, frequency of this infection in women aged over 50 years (age of menopause) is notable and complementary studies are needed.

Microsporidia are eukaryotic, intracellular obligate parasites that widely involve many organisms including insects, fish, birds, and mammals. One of the genera of Microsporidia is Encephalitozoon, which contains several opportunistic pathogens. Since Encephalitozoon spp. are zoonotic and opportunistic pathogens, it is important to find their reservoir hosts; hence, the current study aimed at isolating and identifying Encephalitozoon spp. in the crows by the light microscopy observations and molecular methods. For this purpose, 36 samples were collected by the dropping method; however, due to the low volume of samples, the total samples were collected in a sterile stool container and the polymerase chain reaction (PCR) was performed to detect Encephalitozoon spp. Accordingly, 300-bp bands, specific to Encephalitozoon spp., were observed and by sequencing E. intestinalis was identified. Crows can be considered as the hosts of E. intestinalis.

Introduction: Toxoplasma is a genus of intracellular parasites, which was observed in the many types of mammalians. Toxoplasma protozoa that were detected from many types of rats have the equal phenotypic characteristics but have different clinical symptoms and genotypes. Mostly, domestic animals and rodents are the hosts in the reservoirs in the environment. The objective of this article was to phylogenetic identification of toxoplasmosis in wild-rats of Golestan forest using secretory antigens type 1 gene. Materials and Methods: In this surveillance, the researchers gathered 286 wild rodents from Golestan forest and earned DNA from brain and heart tissues to prove SAG1 gene from these hosts. They split these rodents into four groups and then analyzed the positive samples by the Polymerase chain reaction basis. Results: In this surveillance, they discovered 24% of tissues of the wild-rats were infected for these protozoa. Over 55.8% of tissues belonged to Ratus ratus, 17.6% tissues belonged to Ratus norvegicus, 14.7% of tissues belonged to Mus musculus, 11.7% of tissues belonged to Rombumys opimus, also with attention of phylogenetic analysis, 50 infected rats belonged to genotype ΙΙΙ and 18 infected rats belonged to genotype ΙΙ. Conclusion and Discussion: Conclusion showed that different types of wild rats disseminated this type of infection; also SAG1 molecule was a very important molecule to detect toxoplasmosis in rats of Golestan area.

Background: Toxoplasma gondii is one of the most common protozoan parasites in humans and animals in all countries of the world. The aim of this study was to detect Toxoplasma parasite in the brain of wild rats in Tehran using smear preparation, Giemsa staining, Intraperitoneal injection and oral administration to Souri mice. Materials and Methods: Forty rats were collected from different areas of Tehran. Smears were prepared from rat brains on glass slides and stained using Giemsa. In the second method, a cell suspension was prepared from rat brain and was given orally and injected intraperitoneally into Souri mice. In peritoneal method, peritoneum of the mice was examined for parasites. In oral method, the titer of Toxoplasma antibody in sera of Souri mice was determined using Toxoplasma IgG antibody kit and anti-mouse conjugate of Sigma company. Results: All results were negative in Giemsa staining method. In the second method, the results were negative and no parasites were observed in peritoneum of Souri mice. In oral administration method, after ingestion of suspensions by Souri mice and measuring the IgG titer, 50% of them showed a positive titer after one month. Conclusion: In detection of Toxoplasma gondii, the method of smear preparation on glass slides followed by Giemsa staining, and intraperitoneal injection of brain suspensions to Souri mice are of less value in comparison with oral administration of suspensions and determining the titer of IgG in sera of Souri mice.

Background The aim was to survey the specific factors, which cause to decrease blood feeding of mosquitoes important to succeed vector control. Methods: Larval collection was carried out from fixed and variable breeding places of Yazd County, central Iran in 2009. Autogeny-Anautogeny, Stenogamy-Eurygamy, and blood preference of Culex pipiens were studied using standard mosquito cages blood meal source for Cx. pipiens females considered as the chickens and human and fed females were kept in insectary condition (16:8 L: D, 27±3 °C and 70±10% RH). The data were analyzed using SPSS Ver. 11.5 soft ware. Results: Totally, 96 females’ mosquitoes were tested for Stenogamy versus Eurygamy and 122 for blood preference assay. In the small cages (20 × 20 × 20cm) and large cage (60 × 40 × 60cm), the ability of mating and insemination rates were 60.0 and 67.0%, respectively. In spite of Cx. pipiens fed from sucrose 5%, none of them laying eggs in 60 × 40 × 60 cages during the study. This finding indicated the Anautogeny behavior of this species. This species was found of low tendency to human blood and almost 4 fold fed on chicken. Conclusion: The occurrence of Steno-Eurygamy, Anautogeny, and Ornithophilic behaviors of Cx. Pipiens was noted. More studies need to be carried out about the bionomics of this species to gain more data about the ecophysiological and behavioral characteristics in other parts of Iran.

Since there have been no published molecular studies of Pneumocystis jirovecii isolates from Iranian patients, we investigated the genotypes of such isolates recovered from HIV-infected patients, those undergoing cancer chemotherapy and patients with chronic obstructive pulmonary disease (COPD). P. jirovecii typing, based on ITS1 and ITS2 sequence analysis, was performed on 34 isolates from Iranian immunosuppressed patients. In total, 44 genotypes were detected of which relative to ITS1, eight known genotypes (A, B, C, E, G, H, N and O) and one novel sequence were noted. Eight known genotypes (b, c, e, g, h, i, j and n) were also found with ITS2. The most frequent ITS1 and ITS2 genotypes were E (21/44, 47.7%) and g (22/44, 50%), respectively. From determined haplotypes, the four most frequent ones were Eg (11/44, 25%), Gg (5/44, 11.3%), Gi (4/44, 9.1%), Ei (3/44, 6.8%), and Hg (3/44, 6.8%). Two novel haplotypes (Hb and Hi) were also identified, along with mixed infections as seven (20.5%) patients were found to have more than one haplotype. It is suggested that novel haplotypes in Iranian patients may be generated through sexual recombination within the host.

Background : Blastocystis hominis is an anaerobic, zoonotic protozoan parasite which inhabits the large intestine of humans and a wide range of other vertebrates. It has a worldwide distribution and infects hosts through the cyst of the parasite by contaminated water or food. Its prevalence is related to hygienic culture, season, exposure to animals, and age. A number of studies in the last decade have confirmed its potential pathogenicity, and many gastrointestinal and extra-gastrointestinal signs have been attributed to it. It has unique morphology, life cycle, and reproduction. The aim of this research is to study molecular prevalence of this parasite in the patients referred to Khorramabad laboratories. Materials and Methods: In this study, conducted for the first time in Lorestan province, 511 stool samples were collected from patients in laboratories of Khorramabad. After DNA was extracted using PCR, the samples were examined for the existence of Blastocystis parasite. Results: Out of the 511 samples studied, 33 ones (6.5%) were infected with Blastocystis. Conclusion: The microscopic diagnosis is challenged by morphological characteristics and other intervening factors, and the PCR method, which has higher sensitivity and specification than other diagnostic methods, is recommended. Concerning the prevalence of the parasite, the world has been divided into two parts of developed and developing countries by the researchers, with 10% and 50% prevalence rates respectively. The 6.5% prevalence in the cited population in Khorramabad is in the range of the prevalence in the developed countries. Therefore, the result is justifiable since springs provide most of the drinking water in the region. However, since ranching is a popular occupation in the region, the prevalence rate is alarming. Therefore, following health instructions and appropriate clothing when exposing to livestock are recommended. In addition, the results of this study and similar studies in the developing countries, on the one hand, and the increasing prevalence of 23% in the US as a developed country, on the other hand, have disturbed the previous division.

Populations of Phlebotomus major were examined in two endemic and nonendemic foci of visceral leishmaniasis in Iran. Based on the shape of the aedeagus and ventrally located hairs of coxite and pharyngeal armatures, two morphotypes were found sympatrically in the endemic area of Borazjan. Significant differences in morphometric survey were observed in at least 11 measured characters. The aedeagus of the non-endemic Miyandoab morphotype, and also of a few specimens from Borazjan, is completely parallel throughout its length with a slightly expanded end. Ventrally located hairs of the middle coxite were longer and more compact. It is close morphologically to P. major neglectus (P. neglectus), which was recently recorded from Iran. It is also morphologically similar to P. notus, which has not yet been reported from Iran and needs further investigation. The aedeagus of the morphotype occurring only in Borazjan is narrower in the middle and the hairs are closer to the base of the coxite and are shorter and more outspread, which makes it similar to P. major krimensis or P. neglectus. The two morphotypes occurring sympatrically in Borazjan do not appear to be subspecies and it may be premature to propose them as separate species. Further investigation is needed to clarify the actual status of P. major s. l. in Iran.

Objective To determine the variations in morphological characters of Culex pipiens complex and identify the species complex. Methods A study was carried out from April to October 2009 in Yazd Province, Iran. This study was performed in two randomly selected rural villages in Yazd County using different sampling methods such as larval collection, hand catch, pyrethrum space spray, light trap, and pit shelter collections. The data were analyzed using SPSS software Ver. 16. ANOVA and Tukey's post-hoc test was used to compare the ratios of the samples. Results The average of the DV/D ration was 0.090±0.007 and the range was 0-0.33. The average D/V ratio was 1.07±0.02, with maximum 1.6 and minimum 0.6. The costa and subcosta intersection were observed beyond the bifurcation of R2+3 in most of the specimens. The bifurcations of R2+3 and M1+3 veins were not on one direction in all samples. The range of the R-Cell/R2+3 ratio was 2.42-7.91. The average ratio of cross wing was 2.25±0.04 with a range of 1.36-3.70. Conclusions More populations of Culex pipiens from different areas of Iran need to be studied to gain complete information about the taxonomy and ecology of the species.

Leishmania infantum is the causative agent of infantile visceral leishmaniasis (IVL) in the Mediterranean Basin and, based on isoenzyme typing of the parasite isolated from dogs; this parasite was considered to predominate in the all foci of IVL in Iran. However, based on PCR detection and sequencing of parasite Cysteine Protease B (CPB), only one out of seven sandfly infections in Phlebotomus perfiliewi transcaucasicus was found to be L. infantum in the current investigation. The six other infections were haplotypes of Leishmania donovani, the causative agent of anthroponotic visceral leishmaniasis (AVL) in West Africa and India. The deduced amino acid of the L. donovani haplotype was found to be novel and the shortest CPB protein reported within the Leishmania spp. Circulation of both L. donovani and L. infantum by P. perfiliewi transcaucasicus, in addition to previous data indicating its ability to circulate L. tropica, suggests that this species, like other vectors of VL, is a permissive vector. Finding L. donovani infecting P. perfiliewi transcaucasicus in the area demands extensive and intensive typing of natural Leishmania infections in epidemiological investigations in Iran and the Mediterranean Basin in general.

Cryptosporidium is a world widely distributed parasite which comparatively has a high prevalence in developing countries. The zoonotic potential of some Cryptosporidium species has made the cryptosporidiosis a significant concern to physicians and veterinarians. The occurrence and zoonotic potential of Cryptosporidium species in probable reservoir hosts for man infections was determined by examining faeces of symptomatic and asymptomatic animals. The aim of this study is to screen the presence of Cryptosporidium in fecal sample of free ranging animals in Tehran using Ziehl–Neelsen staining method. The findings indicate that Cryptosporidium are present in 9/50 (18 %) stray cat (Felis catus), 12/50 (24 %) hooded crows (Corvus cornix), 23/180 (12.7 %) rat (Rattus norvegicus and R. rattus) and 1/40 (2.5 %) pigeons (Columba livia). This investigation confirms the potential role of rats, cats, crows and pigeons for zoonotic transmission of human cryptosporidiosis and they must be considered as reservoir hosts which can endanger public health.

Background: Based on the authors’ knowledge, there is no study on the co-infection of opportunistic agents such as Mycobacterium tuberculosis and Pneumocystis jirovecii in the lungs of Iranian patients with immunosuppression. Objectives: The current study aimed to show the rate of co-infection of M. tuberculosis and P. jirovecii in patients with Human Immunodeficiency Virus (HIV). Patients and Methods: Forty-five pulmonary samples were collected from 30 patients with HIV who also infected with Tuberculosis and Pneumonia. All of the patients were admitted to two university hospitals of Mycobacteriology and the Iranian HIV/AIDS research centers. DNA of P. jirovecii was detected using nested-Polymerase Chain Reaction (nested-PCR) assay. Results: All of the patients were male with the mean age of 32.95 ± 7.15 years. The mean of CD4 cell count was 109.25 cell/mm3. Of 30 patients with HIV, three (10%) were co-infected with M. tuberculosis and P. jirovecii. No other causes of pneumonia were found in those three patients and CD4 cell counts less than 50 cell/mm3 was reported. Conclusions: The results of the current study showed a high rate of co-infection of M. tuberculosis and P. jirovecii in the Iranian patients with HIV. As the immune system condition worsened, the probability of occurrence of Pneumocystis Pneumonia (PCP) increased. Therefore, more specific, most rapid and sensitive tests should be utilized for diagnosis of PCP in this group of patients.

The dihydropteroate sulfate (DHPS) gene is associated with resistance to sulfa/sulfone drugs in Pneumocystis jirovecii. We investigated the DHPS mutation rate in three groups of Iranian HIV-positive and HIV-negative patients by polymerase chain reaction-restricted fragment length polymorphism analysis. Furthermore, an association between P. jirovecii DHPS mutations and strain typing was investigated based on direct sequencing of internal transcribed spacer region 1 (ITS1) and ITS2. The overall P. jirovecii DHPS mutation rate was (5/34; 14.7%), the lowest rate identified was in HIV-positive patients (1/16; 6.25%) and the highest rate was in malignancies patients (3/11; 27.3%). A moderate rate of mutation was detected in chronic obstructive pulmonary disease (COPD) patients (1/7; 14.3%). Most of the isolates were wild type (29/34; 85.3%). Double mutations in DHPS were detected in patients with malignancies, whereas single mutations at codons 55 and 57 were identified in the HIV-positive and COPD patients, respectively. In this study, two new and rare haplotypes were identified with DHPS mutations. Additionally, a positive relationship between P. jirovecii strain genotypes and DHPS mutations was identified. In contrast, no DHPS mutations were detected in the predominant (Eg) haplotype. This should be regarded as a warning of an increasing incidence of drug-resistant P. jirovecii strains.

Background: The use of immune suppressive drugs for organ transplant recipients predisposes them to opportunistic infections, especially by fungal agents. Pneumocystis jiroveci, as an opportunistic pathogen, endangers the patients’ life in those with immune system disorders. Early detection of latent Pneumocystis infection in susceptible patients may help choose the optimal treatment for these patients. Objectives: The aim of this study was to identify and determine the colonization of latent P. jiroveci infection among lung transplant recipients. Patients and Methods: This cross-sectional descriptive study was conducted on lung transplant recipients. Bronchoalveolar lavage (BAL) specimens were collected from 32 patients undergoing bronchoscopy. The samples were aseptically homogenized by 10 mM dithiothreitol, and their DNA was extracted. The mtLSUrRNA gene of P. jiroveci was amplified using nested PCR in two stages. Nested PCR was performed using external primers of pAZ-102-E and pAZ102-H followed by using the PCR product of the first stage and internal primers of pAZ-102-E and pAZ102-L2. Results: The genome of P. jiroveci was revealed by a 346 bp PCR product in the initial amplification and a 120 bp product in the nested PCR. The results showed that seven BAL specimens (21.9%) from lung transplant recipients were positive for P. jiroveci. Conclusions: In molecular epidemiology studies, nested PCR has higher sensitivity than PCR. Results of this study support the colonization of P. jiroveci in patients receiving lung transplantation. Patients who are carriers of P. jiroveci are at a higher risk of P. jiroveci pneumonia.