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Background and Objective: Visceral Leishmaniasis or Kala-azar is an important infectious disease in northwestern Iran. Members of the Leishmania donovani complex, L. donovani and L.infantum, are the two main parasites causing the disease in the world. In this study immunophenotype characters such as N-glycosylation, and T-cell and B-Cell epitopes of CPB gene was evaluated in the Leishmania parasites isolated from sandflies of the region. Materials and Methods: Partial of the CPB (702-741 bp) of Leishmania parasites was amplified and sequenced and used for bioinformatics analysis such as test three dimensional (3D) protein structure, N-glycosylation, and T-cell and B-Cell epitopes. Results: In 7 out of 3477 sand flies there was a positive PCR test for systeine protease B, gene. Also according to findings of this study, both agents of kala-azar L. donovani and L. infantum were bing transfered by sand flies of Phlbtomus perfiliewi transcaucasicus in North West Iran. DNA analysis of the CPB gene showed a cytosine insertion at 5’ end of the proofreading frame of the gene resulted in a stop codon (TGA) seven AA further down and hence translation is halted. This caused a short amino acid chain with only 76 AA much shorter than normal CPB peptide with 234-247 AA. This mutation has not been found in the L.infantum strain resulted in a normal CPB peptide. AA analysis showed no N-glycosylation site, T-cell and B-Cell epitopes on the short peptide of the L. donovani strain. Conclusion: This is the shortest CPB peptide chain reported for the L. donovani complex in the literature. This short peptide could have an effect on host-parasite and vector-parasite interactions. Since the CPBs genes have important implication on host–parasite and play key roles in infection and expression of the disease, further studies on the L. donovani parasite and its diminutive peptide necessitate to improve our understanding about the epidemiology of visceral leishmaniasis in Iran.

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Background and Objective: Cutaneous leishmaniases with two forms of rural and urban is the endemic diseases and as a health problem in our country. Identification of parasite species and type of disease is very important for treatment of disease as well as for planning of control program. The microscopic observations by Giemsa-stained smears is the most common laboratory test for the diagnosis of cutaneous leishmaniasis, but the determination of parasite species is impossible and utilization of other ways such as biochemical and molecular methods is required. This study was carried out to determine the parasite species caused cutaneous Leishmaniasis by Nested PCR in Damghan, Iran.

Materials and Methods: This descriptive study was performed on 67 patients with dermal lesions that referred to Damghan health center laboratory in Iran during 2008. The patient's information were recorded in questionnaire. DNA of Giemsa-stained slides from patients was extracted and evaluated by specific primers of kinetoplast DNA using Nested PCR.

Results: Leishmania parasites were observed in 57 patients under light microscope. The 10 patients were infected by other dermal diseases. The PCR result showed the parasite presence in lesions of 57 patients is Leismania major. 54% of patients were male and 46% were female. 72% of the patients were lived in rural areas. 50.9% of disease was observed in over 25 years old patients. Hands were the most common region of ulcer (44.7%). 48% of the patients had one ulcer and the other patients had two or more ulcers. High prevalence (31.6%) of disease was observed in October.

Conclusion: This study showed that zoonotic cutaneous leishmaniasis to be prevalent in this area and Nested PCR method is a sensitive and accurate to leishmania species characterization.

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protozoan of Leishmania genus and in Iran by Leishmania infantum. The protective immune response against VL is cellular immunity through Th1 CD4+, which dominant chemokiens are IL12, IFN- γ  and IL18 and lead to Th1 response. Single nucleotide polymorphism (SNP) on IL-18 gene and its relation to IL18 levels in blood and IL18 function have been studied in many inflammatory diseases such as Behcect’s disease and tuberculosis. According to the important role of IL-18 in immunity against visceral leishmaniasis, this study was conducted to demonstrate the prevalence of genotypes on -607A/C in promoter region of IL-18 gene.

Materials and Methods: This descriptive and cross-sectional study was done on 91 pateints with confirmed VL, 105 healthy sero-negative controls and 78 seropositive controls during 1999-2009. Salting out method was used to extract DNA and ARMS-PCR was used to determine the genotype of -607A/C allele of individuals. Statistical analysis of genotypes was performed using Chi-Square test.

Results: According to the results, -607C/C was the dominant genotype among the groups (35.8%). Distribution of genotypes among groups had not any significant difference. The lowest genotype among healthy sero-positive and patients were -607A/C and -607A/A, respectively. Statistical analysis of distribution of genotypes, did not reveal any significant difference among groups.

Conclusion: The dominant genotypes of VL patients, healthy sero-negatives and healthy sero-positives were -607C/C (38.5%), -607A/C (37.1%) and -607C/C (35.9%) respectively.

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Background and Objective: Epidemiology of Visceral Leishmaniasis is affected by combination quality and interaction of Parasite-Vector-Host and environmental conditions. So, disease cycle related and eco-social factors and environmental risk factors co-analyzing, help to understanding these interactions, prognosis and orientation in disease control and treatment. This study was done to determine the role of nomadic and non-nomadic lifestyle in transmission of Kala-Azar in the Northwest of Iran.

Materials and Methods: In this case-control study, firstly the prevalence of Kala-Azar among people living in selected villages were determined by both serological test (Direct Agglutination Test: DAT) and immunological test (Montenegro Skin Test: MST) in the Northwest of Iran, on the way of Shahsavan tribe travelling in summer/winter quarters villages. Then DAT was conducted on the dogs presenting in those villages during 2006. One year later Seroconversion rate was calculated through collection of the individual’s negative sera and re-analyzing them via DAT. Finally, occurrence of Visceral Leishmaniasis in relation with various involving factors like dog density/abundance and nomadic and non-nomadic lifestyle using Chi-Square test were determined.

Results: Both MST and DAT were significantly higher in the nomadic lifestyle than in the non-nomadic lifestyle (P<0.05). Three values of prevalence (5.5%), seropositivity (2.7%) and seroconversion (2.5%) were higher in nomads than non-nomads. The GIS studies and electronically prepared maps showed that the endemicity and the infection rate are higher in nomads than non-nomads. There were a negative correlation between general distribution of Visceral Leishmaniasis in relation with environmental conditions altitude, mean temperature and rainfall.

Conclusion: This study indicated that Nomadic lifestyle can play as a risk factor in transmission of Visceral Leishmaniasis due to nomads/dog contacting, their entering in the wild cycle of disease and travelling.

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Background and Objective: Several strains of the Echinococcus granulosus have been described based on morphological characters, intermediate host specificity and/or genetic analysis of mitochondrial and nuclear DNA. The aim of this study was to characterize different E.granulosus isolates by using sequences of mitochondrial atp6 gene.

Materials and Methods: In this study, Sixty infected liver and lungs of cattle, sheep and goats were collected from the abattoir of Varamin city-Iran during 2008. Protoscoleces were removed from each fertile cyst and DNA extracted. New and specific primers were designed for two existing genotypes (G1 and G6) of E. granulosus known to occur in Iran and applied in PCR reactions.

Results: The new primers selectively amplified the G1 and G6 genotypes of E. granulosus with specific bands of 708 and 705 bp respectively. The G1 genotype was identified in all fertile cyst samples.

Conclusion: This study showed that the new primer pairs which specifically amplify portions of the mitochondrial atp6 gene of the G1 and G6 strains of Echinococcus granulosus are proper molecular marker for investigating genetic variation in a number of isolates of E. granulosus from a range of hosts (sheep, goats, cattle) in Iran. The result of sequenced samples showed that our sequences were the same as those reported previously for these strains.

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Background and Objective: Zoonotic Cutaneous Leishmaniasis (ZCL) is a parasitic disease which caused by a protozoan belongs to the genus Leishmania. ZCL is of great public health importance in many countries and also in endemic parts of Iran. Leishmania major is the causative agent, Phlebotomus papatasi as the main vector and Rhombomys opimus is the most important reservoir of the disease. Species identification of Leishmania in a large scale of human samples is necessary to conduct a useful program for controlling the disease outspread. This study was done to identify the Leishmania using microscopic and molecular methods in suspected patients of Cutaneous Leishmaniasis by targeting ITS-rDNA gene, Golestan province, Iran. Materials and Methods: 121 smears collected from suspected patients of ZCL, in Eastern region of Golestan province, Iran during 2009-10, stained and examined under a light microscope. DNA of parasites extracted directly from smears and ITS-rDNA gene amplified. Positive samples digested with BsuRI restriction enzyme, according to RFLP method and subsequently the parasite was identified. After sequencing the ITS-rDNA gene, Molecular software was applied for verification of RFLP results. The achieved results were definitely approved by this procedure. Results: 113 out of 121 and 92 out of 121 samples detected as Leishmania positive using microscopic examination and molecular method respectively. All 92 molecular positive samples digested with BsuRI endonuclease and 90 individuals identified as Leishmania major. In order to final verification, 8 samples of Leishmania major sequenced and confirmed by molecular software analysis. Unfortunately, sequences of two samples which were not Leishmania major were not readable, and consequently, these could not be identified. Conclusion: Comparison of obtained sequences of current study with Gene Bank sequences confirmed L.major in human from Northern Iran. Other species of Leishmania were not identified in this investigation but detection of two other samples, which were not L.major, could indicate the role of other Leishmania species causing infection in human in Eastern region of Golestan province, northern Iran. These findings should be considered to improve the disease control programs, which can be led to increase the rate of public health in Golestan province.

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Background and Objective: Cutaneous leishmaniasis is one of the zoonosis disease which is transmitted by sand fly and has been considered as one of the most important health problems in Iran. This study was designed to assess the status of cutaneous leishmaniasis in Gonbad Kavoos, North of Iran. Materials and Methods: This descriptive-analytical retrospective study was performed on the two groups consist of: 1799 patients (995 men and 804 women) referred to health centers of Gonbad Kavoos in Golestan Province North of Iran during 2009-11. Also 278 men and 271 women were selected randomly in one of the villages for the assessment of acute and scar wounds. Patient’s characteristics such as age, sex, habitat, number and sites of ulcer(s), month and years of incidence were registered. Data were analyzed using SPSS-13 and Chi-Square test. Results: From 1799 under care patients, 995 cases (55.3%) were males and 804 cases (44.7%) were females (P<0.05). 1542 patients (85.7%) resided in rural areas, while 257 (14.3%) lived in urban areas (P<0.05). The most frequent age group was 0-9 years old (43.3%) (P<0.05). Hands were the most common sites of ulcer (42.3%) and 37.9% of the patients with one ulser. The highest prevalency of disease was observed in months of October and November (68.4%) 4% and 78.6% of subjects selected from villages were presented with acute wound and scar, respectivley. Conclusion: This study showed that Cutaneous leishmaniasis in 2010-11 was dispersed in Gonbad-Kavoos a city in North of Iran. The endemicity situation of this disease in this area is hypoendemic.

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Background and Objective: Hydatidosis is a chronic, zoonotic worldwide infection which induced by larval stage of Echinococcus worm. This study was done to assessment the pure and B hydatid cyst fluid antigens for the serological diagnosis of human hydatidosis. Methods: In this descriptive laboratory study, infected liver with Hydatid cyst were obtained from Tehran's slaughterhouses to prepare cyst fluid in different stages. After draining and purifying the cyst fluid, it was centrifuged and subsequently concentrated. Specificity and sensitivity of sera samples including hydatidosis (n=60), worm parasites (n=55), fascioliasis (n=35), toxocariasissera (n=20) and negative control (n=35) were tested by Counter Immunoelectrophoresis (CIEP), ELISA and Dot ELISA methods. Results: Specificity and sensitivity of pure antigen of hydatid cyst using CIEP method was 68.9% and 86.7%, respectively. Specificity and sensitivity of B-antigen using CIEP method was 87.8% and 83.3%, respectively. Specificity and sensitivity of pure antigen of hydatid cyst using ELISA method was 76.7%, and 93.3%, respectively. Specificity and sensitivity of B-antigen using ELISA method was 96.7% and 88.3%, respectively. Specificity and sensitivity of pure antigen of hydatid cyst using Dot ELISA method was 83.3% and 100%, respectively. Specificity and sensitivity of B-antigen using Dot ELISA method was 100% and 98.3%, respectively. Conclusion: B-antigen using Dot ELISA method is the most suitable serological test for the diagnoses of hydatid test.

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Background and Objective: Identification of Fasciola species is important. Fascioliasis is one of the important diseases in animals and humans caused by genus Fasciola. This study was done to determine the identification of Fasciola species with RFLP-PCR in animal liver in Gorgan City, northern Iran.
Methods: In this descriptive study, worms were obtained from the livers of infected sheep and cattle in Gorgan slaughterhouse in northern Iran. DNA of worms was extracted with phenol- chloroform method. Fragment of ITS-1 genome was amplified and TasI enzyme was utilized for amplified fragments then 8 samples were sequenced.
Results: A total of 49 Fasciola worms were isolated from infected cattle and sheep. The PCR products of all specimens were affected by the TasI enzyme, and F.hepatica species showed two fragments and F.gigantica species indicated three fragments. The enzyme in F.hepatica species showed a fragment of 151 bp and a fragment of 312, but in the F.gigantica, three fragments were 151, 93 and 219 bp. 36 (73.46%) worms were identified as Fasciola gigantica and 13 (26.53%) worms were identified as Fasciola hepatica. Out of the six infected sheep liver, 22 were isolated from the Fasciola worms, 13 (59.1%) of which were F.hepatica and 9 (40.9%) of them were F.gigantica. Out of the six infected cattle liver, 27 Fasciola worms were identified, all of which were identified as Fasciola gigantica (100%).
Conclusion: This study showed that Fasciola gigantica is the dominant species in infected livers of the cattle in Gorgan city.

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Background and Objective: Echinococcosis garnulosus is a major zoonotic disease, and surgery is the best treatment for it. So far, several chemical scolicidals have been used to prevent protoscolices leakage during surgery, but due to their adverse effects were not well-received. Therefore, using medicinal plants or compounds driven from them as alternatives were taken into consideration. This study was conducted to compare the in vitro effect of Cannabis sativa essential oil with albendazole on protoscolices of hydatid cyst.
Methods: In this descriptive-analytic study, essential oil of C. sativa was prepared from aerial parts of the plant in flowering season in university of Camerino, Italy, and Gas chromatography – mass spectrometry (GC-MS) analysis was performed. Protoscolices were extracted from the livers infected with hydatid cyst and were exposed to different concentrations of C. sativa (1, 2, 5 and 10 μg/ml) for 10, 30, 60 and 120 minutes. The viability of protoscolices was measured by 0.1% eosin staining. Albendazole was used as a standard medicinal drug.
Results: Essential oil of C. sativa at all tested concentrations led to significant mortality in protoscolices which showed a direct relation with increase in concentration (P<0.05). The highest concentration of albendazole after 2 h led to 13.24% mortality rate while, this amount with C. sativa essential oil at 2µg/ml reached to 20.9%, and at 10µg/ml reached to 26.08%.
Conclusion: This study showed that C. sativa essential oil has promising scolicidal effect compared to albendazole and can be suggested as a natural protoscolicidal agent.

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Background and Objective: Cryptosporidium is one of protozoan parasites. Cryptosporidium is important in human public health. This parasite has many species, some of which are common in animal and human hosts. One of the animal hosts of this parasite is pet bird. This parasite causes digestive and respiratory problems in pet birds. This study was performed for molecular identification of Cryptosporidium as a zoonotic pathogen of pet birds.
Methods: In this descriptive laboratory study, fecal samples of 114 cages (50 Passeriformes and 64 Psittaciforms) from all over Isfahan city in Iran were collected by collecting history and after extracting the genome; the Cryptosporidium was detected by specific primers, based on ssrRNA gene.
Results: In 16.66% of the canaries with gastrointestinal symptoms and 4.54% of apparently healthy canaries the ssrRNA gene of Cryptosporidium was detected. In addition, the gene was detected in 10% of cockatiel and 4.16% of budgerigars and in other species of Psittaciforms was not detected.
Conclusion: Pet birds in Isfahan can be considered as a source of Cryptosporidium infection.

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Background and Objective: Dogs are associated with more than 60 types of common diseases with humans, among which parasitic diseases play an important role in public health. This study aimed to estimate the frequency of gastrointestinal helminths fauna among dogs in Gorgan, Iran.
Methods: This descriptive-analytical study was done on 70 dogs (37 male and 33 female) consisting of 40 shelter dogs, 18 pet dogs and 14 guard dogs in Gorgan (Iran) from November 2019 to January 2020. Age, gender and housing places of the dogs were recorded, and dogs' fecal samples were examined for gastrointestinal helminths by flotation method with Sheather's solution.
Results: The eggs of gastrointestinal helminths were detected in 41 dogs (58.6%), including Toxocara canis (29.3%), Echino-taenia (26.8%), hookworms (24.4%), Trichuris vulpis (7.3%) and Toxascaris leonina (12.2%). The highest and lowest rates of infection were related to Toxocara canis (31.8%) in female dogs Trichuris vulpis (5.3%) in male dogs, respectively. Also, in female dogs, 69.7% of the samples were infected with at least one gastrointestinal parasite, and there was a significant relationship between parasitic disease and female gender (P<0.05). Also, the highest infection rate was observed in shelter dogs (61%).
Conclusion: The frequency of gastrointestinal helminths among dogs in Gorgan is high, particularly among female dogs.