Showing papers in "Iranian Journal of Parasitology in 2011"

Isolation of free-living amoebae from sarein hot springs in ardebil province, iran.

Abstract: Background: Free-living amoebae (FLA) are a group of ubiquitous protozoan, which are distributed in the natural and artificial environment sources. The main aim of the current study was to identify the presence of FLA in the recreational hot springs of Sarein in Ardebil Province of Iran. Methods: Seven recreational hot springs were selected in Sarein City and 28 water samples (four from each hot spring) were collected using 500 ml sterile plastic bottles during three month. Filtration of water samples was performed, and culture was done in non-nutrient agar medium enriched with Escherichia coli. Identification of the FLA was based on morphological criteria of cysts and trophozoites. Genotype identification of Acanthamoeba positive samples were also performed using sequencing based method. Results: Overall, 12 out of 28 (42.9%) samples were positive for FLA which Acanthamoeba and Vahlkampfiid amoebae were found in one (3.6%) and 11 (39.3%) samples, respectively. Sequence analysis of the single isolate of Acanthamoeba revealed potentially pathogenic T4 genotype corresponding to A. castellanii. Conclusion: Contamination of hot springs to FLA, such as Acanthamoeba T4 genotype (A. castellanii) and Vahlkampfiid amoebae, could present a sanitary risk for high risk people, and health authorities must be aware of FLA presence.

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

Abstract: 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 Pvalue 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

Seroepidemiological Study of Human Hydatidosis in Meshkinshahr District, Ardabil Province, Iran

Abstract: 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.

Seroprevalence of toxoplasma gondii in sheep, cattle and horses in urmia north-west of iran

Abstract: Background: Toxoplasma gondii is a zoonotic protozoan parasite found worldwide and responsible for major economic losses in most classes of livestock. This study was aimed to determine the prevalence of T. gondii infection in sheep, cattle and horses in Urmia, north-west of Iran, using MAT. Methods: Blood samples of 276 livestock and 26 horses were collected from July 2009 till April 2010. The data were analyzed by the Chi-square, Fisher's Exact and Cochran’s and Mantel-Haenszel Tests. The level of significance was set at P < 0.05. Results: Thirty-three (21.1%) sheep, 2 (1.6%) cattle and 3 (11.5%) horses were seropositive to T. gondii. Analysis showed that sheep were 15 times more likely to be seropositive comparing to cattle also 2 times more likely to be seropositive than horses. Conclusion: This study showed seroprevalence of equine T. gondii infection with a considerable rate in sheep in Urmia, northwest of Iran. More comprehensive studies on livestock toxoplasmosis are required for further analysis of the parasite reservoir for human infection.

Molecular epidemiology of cryptosporidiosis in Iranian children, tehran, iran.

Abstract: Background: Cryptosporidium is a worldwide protozoan parasite and one of the most common causes of infection and diarrhea in humans and cattle. The aim of the present study was determination of subtypes of Cryptosporidium among children with diarrhea in Tehran by sequence analysis of the highly polymorphic 60-kDa glycoprotein (GP60) gene. Methods: Fecal samples were collected from 794 diarrheic children. Initial identification of Cryptosporidium was carried out on stool samples by Ziehl-Neelsen acid-fast staining method. DNA was extracted from positive microscopically samples and Cryptosporidium genotypes and subtypes were determined, accordingly. Results: Out of 794 collected samples, 19 (2.40 %) were positive for Cryptosporidium oocysts. Sequences analysis of GP60 gene showed that 17 (89.47 %) of the positive isolates were Cryptosporidium parvum and 2 (10.52 %) were C. hominis. All subtypes of C. parvum isolates belonged to allele families IIa (6/17) and IId (11/17). The most common allele in all 17 isolates belonged to IId A20G1a (41.18%). A22G1 (IF) subtype was detected in two C. hominis isolates of the children. Conclusion: The predominancy of C. parvum species (specially, IId A20G1a subtype) in current study underlines the importance of zoonotic Cryptosporidium transmission in Iran.

Toxoplasma Infection in Schizophrenia Patients: A Comparative Study with Control Group

Abstract: Background: Schizophrenia is a serious, chronic, and often debilitating neuropsychiatric disorder. Its causes are still poorly understood. Besides genetic and non-genetic (environmental) factors are thought to be important as the cause of the structural and functional deficits that characterize schizophrenia. This study aimed to compare Toxoplasma gondii infection between schizophrenia patients and non-schizophrenia individuals as control group. Methods: A case-control study was designed in Tehran, Iran during 2009-2010. Sixty-two patients with schizophrenia and 62 non-schizophr enia volunteers were selected. To ascertain a possible relationship between T. gondii infection and schizophrenia, anti-Toxoplasma IgG antibodies were detected by indirect-ELISA. Data were statistically analyzed by chi- square at a confidence level of 99%. Results: The sero-positivity rate among patients with schizophrenia (67.7%) was significantly higher than control group (37.1) (P <0. 01). Conclusion: A significant correlation between Toxoplasma infection and schizophrenia might be

Epidemiological studies on echinococcosis and characterization of human and livestock hydatid cysts in mauritania.

Abstract: Background: Echinococcosis/hydatidosis is considered endemic in Mauritania. The aim of this study is to present an epidemiological study on the echinococcosis in man and animals in the Nouakchott region. Methods: The internal organs from livestock carcasses were inspected for research of hydatid cysts. The hydatid fluid was examined for research of the protoscoleces. Dogs were necropsied for the collect of Echinococcus granulosus. Results: In the Nouakchott Hospital, 24 surgical operation of human hydatid cysts have been per­formed, out of which 50% were localised in the lung, 33% in the liver and 17% elsewhere. Then, the incidence rate would be of 1.2% per 100 000 inhabitants in Mauritania. In the dog, the prevalence rate is 14%. The average number of E. granulosus on the whole dogs is 172 and 1227 on the positive dogs. Concerning the livestock, hydatid cysts found in 30.1% of the dromedary, 5.5% of the cattle and 6.5 of the sheep. The fertility rate of hydatid cysts in humans (75%) and camels (76%) was significantly higher than that of sheep (24%) and cattle (23%) ( P <0.0001). Hydatid infestation is characterized globally by the dominance of pulmonary localiza­tions in hu­mans (50%) and camels (72.7%) and in the liver in sheep (76.1%) and cattle (82.3%). Conclusion: The differences between prevalence rates, the fertility of hydatid cysts and diversity sites localization observed in humans and camels of one hand and the sheep and cattle on the other hand, depends possibly the strain(s) diversity of E. granulosus.

Superoxide Dismutase (SOD) Enzyme Activity Assay in Fasciola spp. Parasites and Liver Tissue Extract.

Abstract: Background: The purpose of this comparative study was to detect superoxide dismutase (SOD) activities in Fasciola hepatica, F. gigantica parasites, infected and healthy liver tissues in order to determine of species effects and liver infection on SODs activity level. Methods: Fasciola spp. parasites and sheep liver tissues (healthy and infected liver tissues), 10 samples for each, were collected, homogenized and investigated for protein measurement, protein detection and SOD enzyme activity assay. Protein concentration was measured by Bradford method and SODs band protein was detected on SDS-PAGE. SODs activity was determined by iodonitrotetrazolium chloride, INT, and xanthine substrates. Independent samples t-test was conducted for analysis of SODs activities difference. Results: Protein concentration means were detected for F. hepatica 1.3 mg/ ml, F. gigantica 2.9 mg/ml, healthy liver tissue 5.5 mg/ml and infected liver tissue 1.6 mg/ml (with similar weight sample mass). Specific enzyme activities in the samples were obtained 0.58, 0.57, 0.51, 1.43 U/mg for F. hepatica , F. gigantica , healthy liver and infected liver respectively. Gel electrophoresis of Fasciola spp. and sheep liver tissue extracts revealed a band protein with MW of 60 kDa. The statistical analysis revealed significant difference between SOD activities of Fasciola species and also between SOD activity of liver tissues (P<.05). Conclusion: Fasciola species and liver infection are effective causes on SOD enzyme activity level.

First Detection of Nosema ceranae, a Microsporidian Protozoa of European Honeybees (Apis mellifera) In Iran.

Abstract: Background: Nosemosis of European honey bee (Apis mellifera) is present in bee colonies worldwide. Until recently, Nosema apis had been regarded as the causative agent of the disease, that causes heavy economic losses in apicultures. Nosema ceranae is an emerging microsporidian parasite of European honeybees, A. mellifera, but its distribution is not well known. Previously, nosemosis in honeybees in Iran was attributed exclusively to N. apis. Methods: Six Nosema positive samples (determined from light microscopy of spores) of adult worker bees from one province of Iran (Savadkouh- Mazandaran, northern Iran) were tested to determine Nosema species using previously- developed PCR primers of the 16 S rRNA gene. As it is difficult to distinguish N. ceranae and N. apis morphologically, a PCR assay based on 16 S ribosomal RNA has been used to differentiate N. apis and N. ceranae. Results: Only N. ceranae was found in all samples, indicating that this species present in Iran apiaries.

Epidemiological Aspects of Visceral Leishmaniasis in Baft District, Kerman Province, Southeast of Iran

Abstract: Background : 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. Methods: 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. R esults: 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. Conclusion: It seems that visceral leishmaniasis is being endemic in southern villages of Baft district, southeast of Iran.

Prevalence of Intestinal Parasites in Bakery Workers in Khorramabad, Lorestan Iran

Abstract: Background: Food contamination may occur through production, processing, distribution and preparation. In Iran especially in Khorramabad, 33° 29' 16" North, 48° 21' 21" East, due to kind of nutrition, culture and economic status of people, bread is a part of the main meal and the consumption of bread is high. In this study, the bakery workers were studied for determining of intestinal parasites prevalence. Methods: The study was carried out during September to November 2010 in Khorramabad. All the 278 bakeries and the bakery workers including 816 people were studied in a census method and their feces were examined for the presence of parasites by direct wet-mount, Lugol's iodine solution, and formaldehyde-ether sedimentation, trichrome staining, and single round PCR (For discrimination of Entamoeba spp). Results: Ninety-six (11.9%) stool specimens were positive for different intestinal parasites. Intestinal parasites included Giardia lamblia 3.7%, Entamoeba coli 5.5%, Blastocystis sp. 2.1%, Entamoeba dispar 0.4%, Hymenolepis nana 0.1%, and Blastocystis sp. 0.1%. Conclusion: In order to reduce the contamination in these persons, some cases such as stool exam every three months with concentration methods, supervision and application of accurate health rules by health experts, training in transmission of parasites are recommended.

Performance of antigen b isolated from different hosts and cyst locations in diagnosis of cystic echinococcosis

Abstract: Background: The aim of this study was to assess the performance of Antigen B (AgB) isolated from different Echinococcus granulosus intermediate hosts and from different cyst locations for immu­nodiagnosis of human cystic echinococcosis (CE). Methods: Hydatid cyst fluids were collected from lung and liver cysts of sheep, liver cysts of goats, lung cysts of camels and cattle, and liver cysts of human. AgB was purified from each of these hydatid cysts fluids. Serum samples obtained from 47 pathologically confirmed cases of CE along with 30 sera samples from non-CE patients and 40 sera from healthy individuals were tested by ELISA using AgB prepared from different hosts or cyst locations. Results: The highest sensitivity (97.8%) for diagnosis of CE was seen with AgB prepared from hu­man liver cysts. This maximal sensitivity was followed by AgB isolated from those of sheep liver and lung cysts. The least sensitivity was found with AgB prepared from bovine lung cysts. The highest specificities (97.1%) were observed with AgB isolated from human liver cysts fol­lowed by those of sheep and goat liver cysts while the lowest specificity was seen with AgB iso­lated from bovine lung cysts. In view of the specificities and sensitivities of the different AgB, the best validity was found for AgB prepared from human liver cysts while the least validity was found with AgB prepared from bovine lung cysts. Conclusion: For any AgB-based tests, obtaining of the antigen from one of these sources will signifi­cantly increase the diagnostic sensitivity and specificity of the assay.

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

Abstract: 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

Disseminated Leishmaniasis Caused by Leishmania tropica in a Puppy from Karaj, Central Iran

Abstract: 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 PCRRFLP 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.

In-Vitro Assessment of the Acaricidal Properties of Artemisia annua and Zataria multiflora Essential Oils to Control Cattle Ticks

Abstract: Background: The aim of this study was to investigate the ‘acaricidal effect’ of Zataria multiflora and Artemisia annua essential oils on Rhipicephalus (Boophilus) annulatus.

A Sensitive and Specific PCR Based Method for Identification of Cryptosporidium Sp. Using New Primers from 18S Ribosomal RNA

Abstract: Background: The main goal of the present study was to develop a new sensitive and specific PCR based method for Identification of Cryptosporidium sp. using novel primers from 18S ribosomal RNA. Cryptosporidiosis in high-risk host groups particularly in neonates and immuno-compromised individuals may result in death. To the best of our knowledge this is the first study regarding develop a new PCR based method to diagnose the cryptosporidiosis in Iran. Methods: A total of 850 human fecal samples from patients clinically suspected to cryptosporidiosis and 100 healthy and diarrheic cattle stool specimens were collected. The simplified formol-ether concentration method was carried out for all samples. They were then examined microscopically by modified Ziehl-Neelsen staining method. Total DNA was extracted by QIA amp DNA stool mini kit. PCR and nested-PCR was carried out by using designed primers. Results: Twenty nine cases of cryptosporidiosis infection in human and 30 samples from cattle microscopically were positive. The described primary and nested PCR method could detect all Cryptosporidium positive samples from human and cattle. Regards to suspected negative samples in primary PCR examination, the Nested PCR could approve two more positive results. Furthermore, Nested PCR analysis was able to detect one more case which was negative in both microscopically examination and primary PCR. Specificity of the test was 100%. Sensitivity of Nested PCR in comparison to our gold standard; microscopy after Ridley concentration modified ziehl-Neelsen, was 100 %. Conclusion: Our developed PCR based method by using new primers devised from 18S ribosomal RNA revealed the ability for identification of the Cryptosporidium species such as C. parvum and C. huminis with high specificity and sensitivity.

Apoptosis of Human Lymphocytes after Exposure to Hydatid Fluid

Abstract: Background: Modulation of the immune response is an important strategy by which establishment and growth of hydatid cyst in the internal organs of human is warranted. Induction of apoptosis in the lymphocytes might be a considerable component. This study was designed to evaluate apoptotic impact of hydatid fluid (HF) on human lymphocytes. Methods: Human lymphocytes were treated with hydatid fluid. After 6 hours of exposure, caspase-3 activity, the central enzyme of apoptosis cascade, was measured by fluorometric assay in the HF-treated lymphocytes and control cells. In addition, the expression of Bax (a pro-apoptotic protein) and Bcl-2 (an anti-apoptotic protein) mRNA was assessed by RT-PCR after 12 hours of exposure. Results: Both the ratio of Bax/Bcl-2 mRNA expression and Caspase-3 activity were higher in the HF-treated lymphocytes relative to the control group. Conclusion: Apoptosis could be as a possible mechanism by which Echinococcus granulosus overwhelms host defenses.

Molecular and Seroepidemiological Survey of Visceral Leishmaniasis among Humans and Domestic Dogs in Mazandaran Province, North of Iran.

Abstract: Background: New cases of visceral leishmaniasis (VL) have been reported recently in some parts of Mazandaran Province, north of Iran where the first human case of VL was reported in 1949. This study aimed to determine the present status of Leishmania infantum infection among humans and domestic dogs using serological and molecular methods in central parts of Mazandaran Province. Methods: In this cross-sectional study, blood samples were randomly collected from 402 humans and fortynine domestic dogs throughout 2009 and 2010 in the central part of Mazandaran Province including Semeskadeh and Kiakola districts where recent cases of human visceral leishmaniasis had been reported there. All the collected samples were tested by direct agglutination test (DAT) for the detection of antiLeishmania infantum antibodies as well as convenience PCR assay on whole blood samples for detection of leishmanial infection and identification of Leishmania species. Results: None of 402 collected human (402) and dog (49) blood samples showed anti Leishmania infantum antibodies at titers 1:3200 and 1:320 as cut-off values of DAT, respectively but only 2 of domestic dogs (4.1 %) were found PCR-positive corresponding to L .infantum. Conclusion: This study confirms the circulation of L. infantum at least among domestic dogs and highlights the sporadic pattern of VL in the studied areas. Further investigations regarding to sand flies fauna and wild canines as reservoir hosts of the disease, are recommended.

Epidemiological aspects of canine dirofilariasis in the north of iran

Abstract: Background: Dirofilaria immitis is an important parasite in dog and other carnivores. Our objective was study on incidence and periodicity of heartworm in north of Iran and using other methods for its diagnosis in addition to Parasitology exam. Methods: This survey spanned two years, between 2006 and 2008. Blood samples were collected from 431 stray dogs distributed along north of Iran, the coastal areas of the Caspian Sea. The Knott’s modified test was used for diagnosis of D. immitis and other filariae. Meanwhile, the periodicity of microfilaria in peripheral blood circulation was calculated and the imaging diagnosis techniques of four dogs that had positive results were done. Result: Diagnostic parasitology results indicated that 16.01% of stray dogs were microfilaremic. Two different microfilariae were diagnosed: D. immitis in 13.69%, Dipetalonema reconditum in 1.86% and in 0.46% both of them. There was no statistically significant between infection to fiariae with sex and age of dogs. Also study on the periodicity of the presence of microfilaria in peripheral circulation showed that the highest rate of those was at 1 am and the lowest rate at 12 pm. Radiographic study showed distinctive signs with varied degrees of severity included: Tortuous and enlargement of main and lobar pulmonary artery, pulmonary parenchymal lesions and Right side heart enlargement that confirmed in electrocardiography. Also in echocardiographic images observed short parallel-sided images with the appearance of equal signs that indicated the presence of the heartworm. Conclusion: These results showed that to obtain a reliable diagnosis of heartworm infection, imaging tests could support parasitological exams.

Seroprevalence of Neospora caninum Infection in Dairy Cattle in Tabriz, Northwest Iran

Abstract: Background: The aim of this study was to determine the seroprevalence of antibody to Neospora caninum in healthy and aborted dairy cattle in Tabriz, capital of East-Azarbaijan in northwest of Iran. Methods: In this cross-sectional study serum samples were collected from 266 healthy and aborted Holestein-Feriesisnc cows from September 2008 to August 2009. The sera were analyzed to detect of antibody against N. caninum using the commercially ELISA kit. Results: Seroprevalence of antibody to N. caninum was 10.5% in Tabriz dairy cattle. Also the abortion rate in all cattle sampled was 33.6% but percentage of seropositive aborted cattle was 18.4%. Conclusion: Neosporosis could be one of the possible causes of abortion in dairy cattle in Tabriz and regarding the distribution in dogs as definitive host for the parasite, further studies in dog and cattle are recommended.

Gastrointestinal Helminths of Magpies (Pica pica), Rooks (Corvus frugilegus) and Carrion Crows (Corvus corone) in Mazandaran Province, North of Iran

Abstract: Background: Corvidae is a cosmopolitan family of oscine birds including crows, rooks, magpies, jays, chough, and ravens. These birds are migratory species, especially in the shortage of foods, so they can act like vectors for a wide range of microorganisms. They live generally in temperate climates and in a very close contact with human residential areas as well as poultry farms. There is no available information in the literature concerning the parasitic infections of these three species of corvidae in Mazandaran Province, northern Iran, so this study was conducted to clarify this. Methods: As there are three species of corvid birds in Mazandaran Province, 106 birds including 79 magpies, 11 rooks, and 16 carrion crows were examined between winter 2007 and spring 2008 at post mortem for gastrointestinal helminths. The helminths were drawn and identified morphologically in the Laboratory of Parasitology, Islamic Azad University, Science and Research Branch, Tehran and also partly in the School of Public Health, Tehran University of Medical Sciences, based on the reference books and identification keys like Soulsby, Khalil et al. and Anderson et al. Results: Four species of nematodes, 2 species of cestodes, 1 species of trematodes and 1 species of acanthocephalans were identified in these three corvid species. Conclusion: Five species of the helminths are identified for the first time in Iran, and the acanthocephalan species is new host record for rooks. It is clear that these corvid birds have diverse range of helminths and can act as carriers for infecting the domestic fowls.

Seroprevalence of Toxoplasma gondii Infection in Dogs in Tehran, Iran.

Abstract: Background: Toxoplasma gondii infects a wide range of animals; felines are definitive hosts and other animals including the dogs are intermediate hosts. The aim of this study was to determine the seroprevalence of T. gondii infection in dogs in Tehran, capital of Iran and to investigate possible associated risk factors.

Morphological and Morphometrical Description of Trichostrongylus Species Isolated from Domestic Ruminants in Khuzestan Province, Southwest Iran

Abstract: Backgrounds: Genus Trichostrongylus (Nematoda: Trichostrongylidae) is one of the most impor­tant zoonotic nematodes with wide geographic distribution in the world. The purpose of the present study was to describe morphological and morphometrical characteristics of male Trichostrongylus species, currently prevalent in domestic ruminants of Khuzestan Province , south­west Iran. Methods: Gastro-intestinal organs of 1600 sheep, goats, cattle, and buffalos, slaughtered in Khuzestan Province, southwest Iran, were examined for infectivity with Trichostrongylus spe­cies. For examination and measurements of helminthes, Azo-carmine staining was per­formed, followed by camera lucida drawings of morphological characters and measurements of morphometrical criteria with a calibrated microscope. Using valid nematodes systematic keys, almost all the parasites were identified at the level of species. Results: Overall, 114 animals were found infected with at least one species of Trichostrongy­lus . Considering morphological characteristics of male Trichostrongylus , six species were identi­fied including T. colubriformis, T. vitrinus, T. probolorus, T. capricola , T. longispicu­laris and Trichostrongylus sp . Conclusion: Although, compared to the previous decades, currently Trichostrongylu s is much less prevalent in the domestic ruminants of the study area, but still different species occur in these animals.

Detection and molecular characterization of cryptosporidium species in recreational waters of chaharmahal va bakhtiyari province of iran using nested-pcr-rflp

Abstract: Background: The aim of this study was to detect and characterize Cryptosporidium spp. in water samples collected from recreational ponds of Chaharmahal va Bakhtiyari Province of Iran . Methods: Thirty water samples were collected from November 2009 to May 2010. Each sample contained 10 liters of water. We used the SSU rRNA-based PCR-RFLP technique. Results: Out of thirty samples examined, 6 (20%) were positive for different Cryptosporidium spp. Restriction pattern analysis showed that C. parvum has been the most prevalent genotype, followed by C. hominis and C. canis , respectively. In this area, the higher prevalence of C. parvum compared with other genotypes is consistent with the distribution of cattle. Conclusion: Farm animals, particularly cattle are the main source of cryptosporidial contamination for recreational waters in this area.

Biopathologic Characterization of Three Mixed Poultry Eimeria spp. Isolates

Abstract: Background: Coccidiosis of domestic fowl, caused by species of the Genus Eimeria, is responsible for important economic losses in poultry production. Because different species and/or strains can vary in pathogenicity and other biological parameters, their precise characterization is important for epizootiological studies. Methods: Fifty samples from litter, whole intestinal tract and feces were collected from poultry houses located in different provinces of Iran. One hundred twenty male day-old broiler chicks were challenged with three selected isolates. Data on weight gain, Food Conversion Ratio (FCR), food intake, lesion scoring and shedding of oocysts per gram of feces were recorded and analyzed by the Duncan’s test. Results: In all treatments, the challenged groups had statistically significant lower weight gain than that of unchallenged control group. Isolate three caused the lowest weight gain and food intake and the worst lesion score as well as FCR. Despite originating from close geographical regions for isolates 1 and 2, the difference in biopathologic factors may be either due to different proportion of identified species or the different pathogenicity of the species present in the isolates. Conclusion: The results highlight the importance of considering various species of Eimeria in designing the preventive, control and treatment strategies to prevent coccidiosis in different regions of Iran. Further characterization of each isolate would be the next step to provide a basis for coccidiosis research with well-characterized local isolates.

Development of Sensitive Detection of Cryptosporidium and Giardia from Surface Water in Iran

Abstract: Background: The protozoan parasites Cryptosporidium spp. and Giardia are known to occur widely in both raw and drinking waters. They are two of the causative agents of waterborne outbreaks of gastroenteritis throughout the world. In the present study, a PCR assay and FA were developed for detection of Cryptosporidium oocysts and Giardia cyst in environmental samples. Methods: We have detected Cryptosporidium spp. oocysts and Giardia cysts in seeded and unseeded environmental water samples by PCR method. Water samples were spiked with oocysts (50, 100,300,500) and filtrated with a 1.2-µm pore size cellulose nitrate and follow by DNA extraction and purification by QIAamp DNA mini kit. Nested-PCR assay amplified an 850 bp fragment of 18s rRNA gene specific for Cryptosporidium and 435 bp fragment of glutamate dehydrogenase (GDH) target gene for Giardia. Also many river water from north of Iran, be checked by these methods. Results: Cryptosporidium and Giardia DNAs were detected in seeded water sample and Giardia was detected in all 5 water samples from river in north of Iran by nested- PCR and FA. Also in one river water sample, Cryptosporidium was detected. Conclusion: This protocol is effective for detection of these waterborne parasites in treated and untreated water samples. This study can also serve as a platform for further investigations and research water source

Rapid Detection of Toxoplasma gondii Antigen in Experimentally Infected Mice by Dot- ELISA

Abstract: Background : Toxoplasmosis is a worldwide endemic disease. In congenitally infected infants and AIDS patients, toxoplasmosis causes high rates of morbidity and mortality. In these cases antibody detection is difficult; so detection of parasite or its components could be useful tool for early detection and following treatment of the infection. Methods : Sixty-three BALB/c mice were injected intra-peritoneal with 5×10 3 tachyzoites of Toxoplasma gondii RH strain, nine mice were sacrificed daily for 7 days. Fourteen mice were in­jected with phosphate buffer saline as control group. Dot-ELISA was performed for detection of T.gondii antigen in mice sera and capture - ELISA was done as golden standard assay too. Results : Toxoplasma gondii antigen was detected from day 2 in mice sera ; 22% of mice sera on day 2, 33% on day 3,77% on day 4 and 100% on day 5 till their death on day 7 had shown antigene­mia by dot - ELISA, no positive result was detected in control mice by dot- ELISA. Conclusion: Dot-ELISA is a sensitive method for diagnosis of T. gondii infection in the animal model; also, this technique is more rapid and easy to perform method in comparison with cap­ture-ELISA.

The Effect of Garlic Extract on Expression of INFγ And Inos Genes in Macrophages Infected with Leishmania major.

Abstract: Background: The study was aimed to show the effect of molecular mechanism of Aqueous Garlic Extract (AGE) on expression of IFNγ and iNOS genes in Leishmania major. Methods: Leishmania major promastigotes (MRHO/IR/75/ER) were added to the in-vitro cultured J774 cell line, the cells were incubated for 72 hours. Various concentrations of garlic extract (9.25, 18.5, 37, 74, 148 mg/ml) were added to the infected cells. MTT assay was applied for cellular proliferation. After 72 hours of incubation, supernatants were collected and total RNA was extracted from the infected cells. The express of IFNγ and iNOS genes were studied by RT-PCR method. Results: The colorimetric MTT assay after 3 days of incubation showed cytotoxic effect of garlic extract with an IC50 of 37 mg/ml. In addition, IFNγ and iNOS genes expression by RT-PCR indicated that garlic extract lead to over expression of these genes in J774 cell line infected with L. major. Conclusion: Garlic extract exerts cytotoxic effect on infected J774 cell line. In addition, the hypothesis that garlic can improve cellular immunity with raising the expression of IFN γ and of iNOS genes confirmed.

Identification and Genetic Variation of Fasciola Species from Tabriz, North- Western Iran

Abstract: Background: Fascioliasis is considered as the most important helminthic infection of cattle and sheep. Traditional approaches using morphological and biologic characters cannot cause a cer­tainty in the accurate and precise identification and intra-specific differences of Fasciola spp. In this study, we identified Fasciola species using ITS-1 marker and described genetic variation of each species of the parasite in isolates from Tabriz slaughterhouse in West Azerbaijan Province, north- western Iran. Methods: Overall, 100 samples (50 from sheep and 50 from cattle) morphologically detected as Fasciola worms were studied for identification of Fasciola species by PCR-RFLP method and intra-species variation of the parasite using RAPD-PCR technique. Results: A region of approximately 460bp in all samples was successfully amplified. There were no identifiable variations among the size of PCR products. Two and three fragments in samples correspond to F. hepatica and F. gigantica was seen, respectively, through PCR-RFLP method. No difference was seen in digestion pattern according to host (sheep or cattle). Different types of each species of the parasite was observed using RAPD-PCR technique. Conclusion: We could have an estimate of frequency of F. hepatica and F. gigantic and different genotypes of the parasite in isolates from one locality in north- western of Iran. By extension of such studies in future to other animal hosts (buffalo and goat) and including more regions to sam­pling, the reliability of the results and their application for control programs in zoonotic diseases will be increased.

Gene Diversity of Trichomonas vaginalis Isolates.

Abstract: Background: Trichomonas vaginalis is protozoan parasite responsible for trichomoniasis and is more common in high-risk behavior group such as prostitute individuals. Interest in trichomoniasis is due to increase one's susceptibility to viruses such as herpes, human papillomavirus and HIV. The aim of this study was to find genotypic differences between the isolates. Methods: Forty isolates from prisoners' women in Tehran province were used in this study. The random amplified polymorphic DNA (RAPD) technique was used to determine genetic differences among isolates and was correlated with patient's records. By each primer the banding pattern size of each isolates was scored (bp), genetic differences were studied, and the genealogical tree was constructed by using NTSYS software program and UPGMA method. Results: The least number of bands were seen by using primer OPD8 and the most by using OPD3. Results showed no significant difference in isolates from different geographical areas in Iran. By using primer OPD1 specific amplified fragment with length 1300 base pair were found in only 8 isolates. All these isolates were belonged to addicted women; however, six belonged to asymptomatic patients and two to symptomatic ones. Conclusion: There was not much genetic diversity in T vaginalis isolates from three different