Helminth parasites of Rhombomys opimus from golestan province, northeast Iran

The aim of the study was to determine the helminthic species occurring in great gerbil Rhombomys opimus collected from Maraveh Tappeh, Golestan Province, northeast Iran. During 2010-2011, a total of 77 R. opimus were captured from rural areas of Maraveh Tappeh, Golestan Province, using Sherman live traps and examined for infectivity with any larva or adult stages of helminthic parasites. Overall, 63 R. opimus [81.8%] were found infected with different helminthic species. The rate of infectivity with each species was as follows: Trichuris rhombomidis 31.2%, Trichuris muris 32.5%, Trichuris spp. 10.4%, Syphacia muris 2.6%, Dipetalonema viteae [Acanthocheilonema viteae] 37.7%, Skrjabinotaenia lobata 15.6%, Hymenolepis [=Rodentolepis] nanafraterna 5.2%, and Taenia endothoracicus larva 1.3%. R. opimus is host for several species of cestodes and nematodes in the study area. The high rate of infectivity with D. viteae indicates the susceptibility of these gerbils to this filarial nematode. Synchronous infections occurred up to four species of helminthes in one host

Geographical distribution of Leishmania species of human cutaneous leishmaniasis in Fars province, southern Iran

The goal of this study was to know the identity of Leishmania species responsible of cutaneous leishmaniasis [CL] in Pars Province, southern Iran. Five counties of Shiraz, Firouz Abad, Ghir-Karzin, Farashband and Larestan were prospected. Forty-four patients exhibiting cutaneous lesions were selected. Samples collected on skin le sions were examined both microscopically [after Giemsa staining] and molecularly [after PCR-RFLP]. On the 44 examined patients, 39 exhibit Leishmama sp. by microscopical examination, all confirmed by PCR. For five patients with negative microscopical examination, PCR was positive for three of them. Among these 42 positive samples, 3 [7%] were infected by L tropica and 39 [93%] by L. major. Leishmania major is the most prevalent species in prospected area and L. tropica occurs in Shiraz and Ghir-Karzin counties

Comparative proteomics study on meglumine antimoniate sensitive and resistant Leishmania tropica isolated from Iranian anthroponotic cutaneous leishmaniasis patients

In order to define the protein expressional changes related to the process of meglumine antimoniate resistance in anthroponotic cutaneous leishmaniasis [CL], we performed a comparative proteomics analysis on sensitive and resistant strains of Leishmania tropica isolated from Iranian CL patients. Cell proteins were analysed with 2-dimensional electrophoresis and differentially expressed proteins were identified by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. Image analysis of the matched maps identified 7 proteins that were either over- or down-expressed: activated protein kinase c receptor [LACK], alpha tubulin [X2], prostaglandin f2-alpha synthase, protein disulfide isomerase, vesicular transport protein and a hypothetical protein. The study shows the usefulness of proteomics in identifying proteins that may express differences between sensitive and resistant L. tropica isolates

Epidemiological aspects of visceral leishmaniasis in Baft district, Kerman province, southeast of Iran

Visceral leishmaniasis [kala-azar] is an endemic disease in some areas of Iran. A cross-sectional study was conducted for sero-epidemiological survey of visceral leishmaniasis [VL] in Baft district from Kerman Province, southeast of Iran. Blood samples were collected from children up to 12 years old and 10% of adult population from Baft villages with a multi-stage randomized cluster sampling. In addition, blood samples were collected from 30 domestic dogs from the same areas. All the collected blood samples were tested by direct agglutination test [DAT] for the detection of anti-Leishmania antibodies in both human and dog using the cut-off value of >/= 1:3200 and >/= 1:320, respectively. Parasitological, molecular, and pathological were performed on infected dogs. Chi-square and Fisher exact tests were used to compare sero-prevalence values. From 1476 collected human serum samples, 23 [1.55%] showed anti-Leishmania antibodies at titers of 1:800 and 1:1600 whereas 14 [0.95%] showed anti-Leishmania infantum antibodies at titers of >/= 1:3200. No statistically significant difference was found between male [1.18%] and female [0.69%] sero-prevalence [P=0.330]. Children of 5-8 years showed the highest sero-prevalence rate [3.22%]. Seven out of 30 domestic dogs [23%] showed anti-Leishmania antibodies at titers >/= 1:320. Leishmania infantum was identified in five infected dogs by nested -PCR assay. It seems that visceral leishmaniasis is being endemic in southern villages of Baft district, southeast of Iran

Comparison of microscopical examination and semi-nested multiplex polymerase chain reaction in diagnosis of Plasmodium Falciparm and P. vivax

We compared light microscopy examination and a semi-nested multiplex PCR [SnM-PCR] assay in endemic areas of the Islamic Republic of Iran. A total of 68 individuals with malaria-positive and suspected malaria symptoms were included in the study. Giemsa-stained thick blood films were examined under a light microscope for malaria parasites in 100 and 200 fields. DNA was extracted from blood samples and SnM-PCR based on the amplification of the small subunit ribosomal RNA [ssrRNA] gene sequences was applied. Microscopical examination showed that 48.5% [33.8% P. vivax and 14.7% P.falciparum] and 50% [35.3% P. vivax and 14.7% P.falciparum] of the samples were positive in 100 and 200 fields respectively. SnM-PCR showed the same results as the 200 field microscopy

[Establishment and integration of a visceral leishmaniasis surveillance system in the primary health care system and its evaluation, Meshkin-Shahr District, Iran]

Visceral leishmaniasis is a systemic parasitic disease with a high fatality rate in under-5-year-old children. The disease is endemic in some parts of Iran, particularly in the north-west region. In 2001 a visceral leishmaniasis [VL] surveillance system was established for children aged>/=12 years in the primary health system in Meshkin-Shahr District, Ardebil Province, situated in the north-west of Islamic Republic of Iran. All cases with clinical signs and symptoms of VL and confirmed positive by the direct agglutination test [DAT] were referred for physical examination and treatment. The mean annual incidence of VL decreased significantly from 1.88 per 1000 children before [1985-2000], to 0.77 per 1000 child population after [2001-07], the intervention. In the control area with no surveillance, it increased from 0.11 to 0.23 per 1000. Early detection of VL using serological tests and timely treatment of cases can decrease the mortality and morbidity rates of VL in endemic areas

Disseminated leishmaniasis caused by Leishmania tropica in a puppy from Karaj, central Iran

A 5-month old puppy with muco-cutaneous lesions in the chin, around lips and eyes was examined physically and microscopically for leishmaniasis. Muco-cutaneous lesions containing a large number of amastigotes of Leishmania spp. were observed. Amastigotes were also detected in liver and spleen of the puppy. The animal was positive with Dipstick rK39 kit and high level of anti-Leishmania antibodies was detected by direct agglutination test [DAT]. DNA, Using PCR-RFLP technique extracted from cultured Leishmania promastigotes and L. tropica was identified. This is the first report of concurrent mucosal and visceral involvement of L. tropica in a puppy from Iran

Neospora caninum and leishmania infantum co-Infection in domestic dogs [canis familiaris] in Meshkin-Shahr district, northwestern Iran

Mediterranean visceral leishmaniasis [MVL] is an infectious disease that affects both human and animals. Domestic dogs [Canis familiaris] are principal reservoir hosts of MVL caused by Leishmania infantum. Dogs are definitive hosts for Neospora caninum and a risk factor for infecting intermediate hosts. The immunosuppression caused by visceral leishmaniasis [VL] can promote the occurrence of co-infections with other agents such as neosporosis. This study aimed to determine the frequency of co-infection of the both protozoan parasites in the endemic areas of VL from Meshkin-Shahr District, north-west of Iran. Altogether, 171 serum samples were collected from domestic dogs of Meshkin- Shahr District by multistage cluster sampling from October 2008 to August 2009. The collected serum samples were tested for the detection of simultaneous infection of L. infantum and N. caninum using direct agglutination test [DAT] and indirect ELISA, respectively. Of the 171 domestic dogs, 27 [15.8%] and 52 [30.4%] were showed antibodies against L. infantum and N. caninum, respectively. Simultaneous infections of N. caninum and L. infantum was found in 16 [9.4%] of the dogs. In VL-positive and VL-negative dogs, N. caninum infection was found in 59.3% and 25.0%, respectively. A statistically significant difference was found between VL-positive and VL-negative dogs with N. caninum infection [P= 0.001]. These findings indicate that Meshkin-Shahr District in northwestern Iran is an active focus of canine visceral leishmaniasis [CVL]. Neospora caninum and L. infantum co-infection is prevalent in the area and infection by L. infantum seems to enhance susceptibility to N. caninum infection in domestic dogs

Integrated visceral leishmaniasis surveillance system in primary care for children in Meshkin-Shahr district, north-western Islamic Republic of Iran

In 2001 a visceral leishmaniasis [VL] surveillance system was set up for children aged

Protein profiling on meglumine antimoniate [Glucantime [R]] sensitive and resistant L. tropica isolates by 2-dimentional gel electrophoresis: a preliminary study

Glucantime[R] is the first- line drug for the treatment of all forms of leishmaniasis. Unfortunately, the prevalence of parasites becoming resistant to Glucantime[R] is increasing in several parts of the world including Iran. As protein is the most important target for drugs in response to a variety of signals including drugs so, it seems expression protein patterns in sensitive and resistant Leishmania parasites could greatly help us about the mechanisms of responses to antileishmanial drugs. In this study, we used 2-dimentional gel electrophoresis [2-DE] method to determine protein expression profiles between drug [Glucantime[R]] sensitive and resistant Leishmania tropica isolated from Iranian an throponotic cutaneous leishmaniasis [ACL] patients. We used from the two confirmed genetically of Glucantime[R] sensitive [Mash-4] and resistant [Mash-927] field strains of L. tropica, isolated from ACL patients in north eastern Iran. The two Leishmania isolates were cultured, promastigotes were harvested followed by protein extraction using TCA/Aceton to study protein profiling, 2-DE was done and gels stained with silver nitrate. At least 2236 distinct protein spots were detected. Twelve spots out of them, showed significant changes in expression in resistant compared to sensitive isolates. Of these, 11 protein spots were up- and one was down-regulated. This preliminary study has showed that a number of proteins differentially expressed in drug [Glucan-time [R]] resistance L. tropica and probably the role of these proteins are increasing the parasite resistance against the drug and delay in cell death

Molecular identification of leishmania species isolated from human cutaneous leishmaniasis by RAPD-PCR

Characterization of Leishmania parasites is necessary for epidemiological objectives such as documenting the distribution of predominant species and designing appropriate control measures. In this study, we aimed to identify Leishmania species isolated from cutaneous leishmaniasis [CL] patients, using RAPD-PCR method. This descriptive, cross-sectional study was designed against 20 Leishmania spp. which were confirmed by parasitological examination, isolated from 30 suspected cutaneous leishmaniasis patients, referred to Health Centers of Kermanshah Province from August 2006 to December 2007. All desirable samples were characterized by RAPD-PCR method using five selected oligoprimers [AB1-O7, A4, 327, 329 and M13]. Electrophoresis patterns from each isolate were compared with reference strains of L. tropica, L. major and L. infantum. Eighty nine percent and 11% of isolates were similar to L. major and L. tropica reference strain, respectively. Five of the isolates were identified as L. major using RAPD-PCR, induce ulcers at the base tail of Balb/c mice, 4-6 months after inoculation. L. major is dominant in the studied areas and it seems that some parts of the Kermanshah Province to be probably considered as zoonotic cutaneous leishmaniasis areas in the middle west of Iran

Variation of the chloroquine resistance transporter [crt] gene in chloroquine-resistant and chloroquine-sensitive plasmodium berghei

The emergence and spread of chloroquine resistant Plasmodium falciparum in the world stimulated some investigators to consider different aspects of chloroquine resistance in human and rodent Plasmodia. Using animal Plasmodia, particularly primate and rodent Plasmodia can be useful model for human Plasmodia studies. In this study we have tried to consider and compare the sequence of chloroquine resistance transporter [crt] gene among chloroquine-resistant and chloroquine-sensitive strains of Plasmodium berghei. This experimental study was performed at the Malaria Laboratory of School of public health. DNA was extracted from two strains of P. berghei which their resistance and sensitivity had been demonstrated in mice with treatment by chloroquine. By using specific primer for crt gene some parts of this gene were amplified by PCR, and obtained fragments were then sequenced and compared. There were considerable differences in crt gene between two strains. Sequenced 1212 bp of crt gene fragment in the two strains showed 43 differences at nucleotides level and 16 differences in presumed coding amino acids. crt can be addressed as a considerable gene which involves in induction of resistance to chloroquine in P. berghei, as P. falciparum. The results increased such a promise that considering crt gene in chloroquine-sensitive and chloroquine-resistant P. berghei can prepare suitable and helpful fields for more understanding the molecular aspects of chloroquine-resistance in Plasmodia and reversing the effectiveness of 4-aminoquinolines particularly chloroquine for treatment of drug resistant Plasmodia

[Seroepidemjological survey of visceral leishmaniasis by direct agglutination test in bojnoord district, north Khorasan province in 2007]

This study aimed to determine of the seroprevalence of visceral leishmaniasis to design a prevention and control program in Bojnoord district. This was a descriptive and cross sectional study with multistage random sampling method. The study was carried out on children up to 12 years old and 10% of adult's population in 8 rural areas of Bojnoord district in 2007. In total, 1608 blood samples were collected to detect anti-Leishmania antibodies. All the samples were tested by direct agglutination test [DAT]. In order to determine Leishmania species, necropsy was performed on four suspected dogs in the studied areas. The species of Leishmania was determined by RAPD-PCR and PCR-RFLP methods using to amplify the ribosomal internal transcribed spacer 1 [ITS1]. Thirty and eight [2.36%] out of 1 608 collected blood samples had anti- Leishmania antibodies at titer 1:800 and nine [0.56%] were just positive at 1:3200 by DAT. There was no statistically significant difference between female and male seroprevalence [p>0/05]. The seoprevalence in children <=12 years old compared to individuals greater than 12 years old did not show any statistically significant [p>0/05]. Amastigotes were observed in all 4 necropsied dogs. The species of Leishmania isolated from 2 dogs, was determined as L.Infantum. Their ITS1 sequences were registered with Accession numbers EU810776 and EU810777 in NCBI. These findings showed that visceral leishmaniasis has been circulated with low endemicity in Bojnoord district. Therefore an appropriate monitoring system is needed for health services in this area

Emergence of Cutaneous Leishmaniasis due to Leishmania major in a New Focus of Southern Iran

Reports from the health center of Kerman Province, southern Iran showed an increasing of cutaneous leishmaniasis cases in Orzuieh Rural District, southwest of the province in 2003. The report encouraged the team to carry out an epidemiological survey in the district during 2003-2004. The objectives were to determine the ecology of sand flies, potential reservoir hosts and human infection. A total of 1075 sand flies were collected by sticky traps and 7 species of sand flies were identified. They comprised 3 species of the genus Phlebotomus [P. papatasi, P. mongolensis and P. bergeroti] and 4 species of the genus Sergentomyia [S. sintoni, S. clydei, S. tiberiadis and S. Baghdadis]. P. papatasi was the predominant species of the genus Phlebotomus in indoors [90.3%] and outdoors [50.2%]. Susceptibility tests on P. papatasi with DDT 4%. Showed that the species was susceptible to this insecticide. A total of 13 rodents consist of Tatera indica [76.9%] and Nesokia indica [23.1%] were collected. A study of prevalence among 2441 inhabitants in four villages showed a rate of 1.1% for active lesions and 10.4% for scars during November- December 2003. In a separate study examination of 1662 school children aged 6-12 years old showed 1.14% for ulcers and 14.7% for scars at the same time. The Leishmania parasites were isolated from man and characterized as Leishmania major using RAPD-PCR method. It seems that cutaneous leishmaniasis due to L. major [CLM] has been prevailed in the district

Leishmania tropica: another etiological agent of canine visceral leishmaniasis in Iran

Dogs have been previously reported to be reservoirs of Leishmania infantum as the etiological agent of human visceral leishmaniasis in Iran. We report a case of canine visceral leishmaniasis [VL] caused by L. tropica from the north- west of Iran where human visceral leishmaniasis is endemic. The canine VL was initially screened by dipstick rK39 and direct agglutination test, then the dog was dissected and obtained samples were examined by parasitological [direct exam, cultivation] and molecular techniques [RAPD-PCR and RFLP-PCR]. Leishmania parasites were found in spleen and liver of the dog. The serological tests for the detection of specific anti-leishmania antibodies showed positive results. L. tropica as another agent of canine VL was determined

Application of direct agglutination test [DAT] for the diagnosis and seroepidemiological studies of visceral leishmaniasis in Iran

Visceral leishmaniasis [VL] is one of the most important parasitic diseases which is endemic in different parts of Iran. Serological studies were conducted by direct agglutination test [DAT] on 12144 human serum samples, collected from four geographical zones of Iran. Sero prevalence, geographical distribution, clinical signs and symptoms for human visceral leishmaniasis based on DAT for the period of 2002 through 2005 were determined. From 516 kala-azar cases detected: 50.6% were from Meshkin-shahr and Moghan districts in Ardabil Province, northwest of Iran and 49.4% were detected from other areas of Iran. In physical examination of seropositive cases, which were detected by DAT with anti-leishmanial antibodies at titers of 1: 3200 to 1: 102400, almost 50% of suspected individuals showed the classical kala-azar signs and symptoms. Predominant signs and symptoms in 233 hospitalized patients with anti-Leishmania antibodies at 1:3200 and higher, were fever [88.0%] and splenomegaly [84.5%]. Statistically significant difference was found between males [58%] and females [42%] [P< 0.01]. Moreover, 93.6% of the VL patients were < 5 yr of age, and 6.4% were older than 5 yr that this difference was statistically significant [P< 0.01]. From 1383 serum samples collected from domestic dogs in the villages that are known as endemic foci of human leishmaniasis, 152 [11.0%] were positive by DAT [>/= 1:320]. Parasitological and serological examinations that were performed in 30 wild canines showed that 10% of these animals were infected by L. infantum. L. infantum Lon49 is the principal agent of the disease in human as well as animal reservoir hosts in different parts of Iran. For the first time in Iran, L. tropica isolated from both skin lesions in the face and bone marrow aspiration in a HIV+ man who co-infected with VL as well as in an infected dog from Ardabil Province

Characterization of Leishmania infection in rodents from endemic areas of the Islamic Republic of Iran

Between 1991-2000, Leishmania species were isolated and characterized by isoenzyme and molecular analysis from rodents caught in various parts of the Islamic Republic of Iran. In areas endemic for cutaneous leishmaniasis, parasites were observed by direct microscopy in smears from 18.6% of 566 specimens. L. major was isolated from 4 species: Rhombomys opimus, Meriones libycus, Tatera indica and Mer. hurrianae. L. turanica was isolated from R. opimus for the first time in this country. In endemic areas of visceral leishmaniasis, parasites were observed in liver and spleen from 13.7% of 504 rodents. Two species were positive on culture; promastigotes isolated from Mer. persicus were characterized as L. donovani zymodeme LON50 and from Mesocricetus auratus as L. infantum LON49

Identification of leishmania sepsis isolated from human cutaneous leishmaniasis, using r and om amplified polymorphic DNA [RAPD-PCR]

This cross-sectional study was designed to isolate of Leishmania spp from cutaneous leishmaniasis patients and characterized them by RAPD-PCR technique. Eighty- seven Leishmania isolates from 112 samples were collected from cutaneous leishmaniasis [CL] patients who referred to Mashhad Health Centers from August 2002 to May 2004. Desirable samples [87 isolates] were characterized by RAPD-PCR method using four selected oligoprimers. Electrophoresis patterns from each isolate were compared with reference strains of L. major, L. tropica and L. infantum. The results showed that 94.2% and 5.8% of isolates were similar to L.tropica and L.major reference strain, respectively. Four isolates that were determined by RAPD-PCR as L.major, could produce ulcer at the base tail of BALB/c mice, 4 - 12 weeks after inoculation but none of L. tropica isolates produced any lesions at the site of injection in the animals. The results indicate that L. tropica species are dominant in the studied areas of Mashhad city and RAPD-PCR technique is a suitable tool for Leishmania characterization in epidemiological studies