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Glucose phosphate dehydrogenasis (G6PD), the most prevalent enzymatic disease in humans, exists in south-eastern Iran. The geographic correlation of its distribution with the historic malaria endemic suggests that G6PD has increased in frequency as a result of natural selection by malaria. Based on studies, there is a controversy in terms of different analytical methods in terms of resistance to malaria. Fifty malaria patients and 50 healthy individuals from several cities south-east of Iran were included in the study and after obtaining consent, blood samples were taken from them. G6PD enzyme deficiency was investigated using a fluorescent stain test. The age, gender, and nationality of malaria patients were also assessed. The results were analyzed using SPSS software and appropriate statistical tests, and the value of P0.05 was considered significant. The results showed that in malaria patients only one person had G6PD deficiency, while this number was six in the control group, which is significantly higher than the malaria group (P0.05). Age group 27-42 years, men and people with Iranian citizenship also showed the highest incidence of malaria. Based on the results, it can be concluded that G6PD enzyme deficiency causes resistance to malaria and the frequency of this enzyme deficiency in malaria patients is significantly lower than in other people.

Background Cryptosporidiosis has a worldwide distribution, and is the commonest cause of diarrhea in children and immune compromised individuals. Since there is no data available on the prevalence of Cryptosporidium species (sp.) in Zabol city, thus this study was carried out to assess the disease prevalence and related factors influencing the disease. Materials and Methods In this cross-sectional study, 200 fecal specimens were collected from children referred to the Central or hospital labs in Zabol city, South East of Iran, during April 2014 to August 2016. Fecal examination was performed by staining with Ziel-Neelsen acid-fast to find oocysts of the parasite. The children were grouped according to the age, gender, kind of water supplies, and diarrheic and non-diarrheic condition. Data were evaluated using SPSS version 13.0 software. Results Among the children referred to the Central laboratory, 200 fecal samples from different age groups were collected. The prevalence of Cryptosporidium species was 9.7% which was higher in children under 4 years. There was a significant relationship between sources of water supply and diarrheic children infected with Cryptosporidium (P

Background and Objectives: Leishmaniasis is a public health problem caused by the protozoan Leishmania. Pentavalent antimonials are currently used for treatment of leishmaniasis, but they have serious side effects. Nerium oleander L. has been used in traditional medicine due to its various health-protective properties. This study aimed to investigate anti-leishmanial activity of N. oleander L. leaves extract against Leishmania major promastigotes and amastigotes in vitro. Methods: L. major promastigotes were cultured in RPMI 1640 medium supplied with 10% fetal bovine serum. Different concentrations were prepared from the extract and added to L. major promastigotes seeded in 96-well plates. Viability percentage was evaluated by direct counting and MTT assay after 24, 48 and 72 hours. To investigate the cytotoxic effect of N. oleander L. on L. major amastigotes, the plant extract was added to amastigotes cultured in intraperitoneal macrophages. The mean number of amastigotes was calculated by direct counting after 24 and 48 hours. Results: All concentrations of the extract significantly reduced the viability of promastigotes when compared with the controls. Half-maximal inhibitory concentration was estimated to be 22.21 µg/ml after 24 hours. Percentage of cytotoxicity in amastigotes exposed to 20 μg/ml of the extract was 53.61% and 53.27% after 24 and 48 hours, respectively. In addition, percentage of cytotoxicity in amastigotes exposed to 80 μg/ml of the N. oleander L. extract was 53.77% and 55.48% after 24 and 48 hours, respectively. Conclusion: The N. oleander L. extract exerts anti-leishmanial activity on L. major promastigotes in a time- and dose-dependent manner. Keywords: Leishmania major, Nerium.

Introduction: Toxoplasmosis is a parasitic disease caused by the protozoan, Toxoplasma gondii. In people with healthy immune system, generally, the infection is asymptomatic, but for the people with immune deficiency severe disease can be observed in case of the acute infection or activation. Determination of prevalence of anti-Toxoplasma antibodies is indicative of exposure to the infection in on-dialysis patients, since they are susceptible of many diseases. Objectives: This study aimed at determination of prevalence of anti-Toxoplasma IgM and IgG in on-dialysis patients comparing to the healthy people in Sistan area. Patients and Methods: This study was of a case control type. Subjects included all of the on-dialysis patients in Sistan as a case group, 76 persons, and healthy people as the controls, 76 persons, who referred to Imam Khomeini hospital of Zabol and Seyyed-Al-Shohada hospital of Zahak, from which 85 were male and 67 were female. Simple sampling was used. The materials were demographic information and enzyme linked immunosorbent assay (ELISA) was used for determination of anti-Toxoplasma IgM and IgG. The obtained data were then analyzed with SPSS v. 16 using chi-square test. Results: Anti-Toxoplasma gondii IgG was observed in 56 (73.7%) cases of the on-dialysis patients and 33 (43.4%) cases of healthy individuals. Anti-Toxoplasma IgM was not detected in the patients in either groups. The difference in IgG antibodies among two groups was statistically significant (P < 0.001) Conclusion: The prevalence of IgG antibodies against Toxoplasma in the on-dialysis patients is higher compared to healthy group, thus these patients are at the higher risk of infection with Toxoplasma gondii.

Background: The aim of the present survey was to assess thr seroepidemiologic and parasitological aspects of Toxocara canis infection in children under 14 yr old. Methods: Overall, 963 sera were collected from children in the Sistan and Baluchistan Province, Southeast of Iran during the period from Sep 2015 to Jun 2016. IgG antibody against T. canis in the subjects’ sera was evaluated using the commercial ELISA kit. Results: Anti-Toxocara IgG were detected in the serum of 17 (1.7%) of the participants. In the examined children, the highest presence of anti-Toxocara antibodies was 2.1% (9/418) in 6-10-yr olds, which was higher than other age groups (P

Background: The detection of Fasciola species in various geographical regions is essential for health policymaking. Here, we aimed to identify livestock (cattle and sheep) related Fasciola genotypes by restriction fragment length polymorphism PCR. Methods: Seventy adult Fasciola flukes were collected from 70 infected livers of 35 cattle and 35 sheep slaughtered in Zabol abattoir, outh-east Iran (Jan-Jul 2017). Fasciola species were determined based on molecular features. For molecular detection, Fasciola ITS1 region was amplified and sequenced. A 700 bp fragment was amplified. These were digested with RasΙ enzyme. F. hepatica specific fragments were 47, 59, 68, 104, and 370, while those related to F. gigantica had 45, 55, 170, 370. Results: The two main species of F. hepatica and F. gigantica are responsible for fasciolosis in sheep and cattle in our region. From 35 Fasciola isolated from cattle, 3 and 32 were F. hepatica and F. giagantica respectively. From 35 Fasciola isolated from sheep, 4 were F. hepatica and 31 were F. gigantica. Conclusion: All Seventy Fasciola samples from two different hosts (cattle and sheep) were identified as either F. hepatica or F. gigantica by PCR-RFLP. Genotypic variability of Fasciola species was high in our region. It is recommended to assess molecular variation of Fasciola isolates in other host livestock.

Background: Cryptosporidium parasite is the cause of human gastroenteritis and other cold and warm-blooded animals that have been widely distributed throughout the world. Genetic information on opportunistic pathogens in immunocompromised patients leads to an increase in the information on epidemiology, patient care, patient management, and rescue. In Iran, infection to Cryptosporidium spp. has been reported, yet only molecular genes can differentiate species and genotype discrimination of the cyst. The molecular assays indicated that Cryptosporidium parvum is the most common species found in Iran, followed by C. hominis. Objectives: The present study aimed at determining the genetic diversity of Cryptosporidium (C.) in children with diarrhea using the PCR-RFLP method and SSU gene. Methods: In this study, stool specimens were collected from 182 children with diarrhea referring to Zabol hospitals. Slides and shitter procedure were done and Ziehl-Neelsen stain was observed directly; an examination was made to identify the parasite, and PCR-RFLP were eventually performed on DNA extracted from the isolates. Results: Out of 182 stool specimens, 27 isolates were identified as Cryptosporidium, using Ziehl-Neelsen stain method, of which 17 and 10 isolates were respectively reported to be C. parvum and C. hominis after the molecular examination. Conclusions: Both human and cattle genotypes are seen in children with diarrhea, yet since the dominant species is C. parvum, zoonosis is more common than human transmission and human-livestock contact is considered as the most important source of human contamination.

Background: Cutaneous leishmaniasis is one of major public health problem in many countries. Pentavalent antimonite components as first line treatment of cutaneous leishmaniasis has many limitations and side effects. Objective: The aim of this study was to investigatethe cytotoxic effect of methanolic extract of Terminalia chebula Retz fruit on the Leishmania major in vitro. Methods: Terminalia chebula Retz fruit extract with 80, 125, 250 and 500 µg/ml concentrations with leishmania promastigotes were added to 96-well plate as 4-time repeated and incubated in 21 ˚C. Viability percent of promastigotes was estimated by direct counting and MTT assay; after 24, 48 and 72 hours. Terminalia chebula Retz extract with80 and 250 μg/ml were added to infected macrophages. The mean of amastigotes in each macrophage was obtained after 24 and 48 hours. Statistical analysis of data was done by SPSS software and one-way ANOVA test. Results: viability percent of promastigotes in groups treated by maximum and minimum concentrations (500 and 80 μg/ml) of Terminalia chebula Retz after 24 hours was 19.01% and 98.65%, respectively.Viability of this groups after 48hours was 7.81% and 97.26% and after 72 hours it was 31.61% and 97.26%, respectively. IC50 for 24 hours was 114.10 µg/ml. Cytotoxicity of Terminalia chebula Retz extract (80 and 250 μg/ml) in amastigotes after 24 hours was 40.35% and 29.99%; and after 48 hours was 45.21% and 41.11%, respectively. Conclusion: extract of Terminalia chebula Retz fruit has cytotoxic effect on the promastigotes and amastigotes of L. major in vitro.

Background: The purpose of this study was seroepidemiological and parasitological assessment of Toxocara canis infection in children and dogs in Zabol and Chabahar, Iran. Methods: This study was a descriptive-analytic study with a simple random sampling of children under 14 yr old, referring to urban, rural, and tribal laboratories of Zabol and Chabahar, Sistan and Baluchestan Province, Iran in 2016. Demographic data, clinical, and laboratory conditions of patients were collected through interviews, questionnaires, and blood count measuring. The prevalence of IgG antibodies against T. canis was assessed by ELISA. T. canis eggs in dogs (as the original host) were also assessed by examining animal feces. Then the data were analyzed using SPSS 19 software and descriptive statistics, chi-square and ANOVA statistical tests. Results: Totally, 364 patients were enrolled, of which 51.6% were female and mean±SD age of participants was 7.2 (±3.7) yr. IgG antibodies against T. canis was observed in 3.8% of cases. A significant association was found between the seroprevalence of T. canis and eosinophil (P=0.003) and red blood cell count (P=0.04). We also found a significant association between serological prevalence of T. canis and demographic parameters, such as city of residence (P=0.003), gender (P=0.04), consumption of vegetables (P=0.01), and the living place (P=0.04). Mean antibody titration was 2.2 ±1.1, with statistically significant difference among age groups (P=0.001). In addition, T. canis infection was positive in 27.5% of dogs living in the study areas. Conclusion: High risk of infection represented in patients referring to laboratories of Zabol and Chabahar. In addition, given the fact that dogs are the final hosts to transfer Toxocara infection to humans, this study emphasizes the need to control the population of stray dogs in the region to prevent the development of disease in the human society.

OBJECTIVE Acanthamoeba is one of the most abundant free-living amoebas that is widely distributed in natural and artificial environment resources. Acanthamoeba pathogenic genotypes cause chronic human diseases including amoebic keratitis and granulomatous amoebic encephalitis. The aim of this study was to determine and identify Acanthamoeba genotypes residing in pools and stagnant water in ponds in Sistan region in southeast Iran. This descriptive study was conducted at the Parasitology Laboratory, School of Medicine, Zabol University of Medical Sciences. METHODS In this descriptive study, 93 water samples were collected from pools and ponds in Zabol, Zahak, Hirmand, Hamoon, and Nimrooz in Sistan region. Samples after filtering through 0.45-μm nitrocellulose paper filters were cultured in a 1.5% non-nutrient agar medium enriched with heat-killed Escherichia coli. Polymerase chain reaction (PCR) was conducted using specialized primers for detecting the genus Acanthamoeba. The sequencing of positive samples was used for determining Acanthamoeba genotypes. RESULTS From 82 free-living amoeba positive culture samples, 38 isolates were confirmed to belong to the genus Acanthamoeba by PCR. On sequencing, 34 samples (89.47%) belonged to the T4 genotype, three (7.9%) to the T5 genotype, and one (2.63%) to the T3 genotype. CONCLUSION All genotypes found in this study are potentially pathogenic. The T4 genotype is the main genotype of Acanthamoeba responsible for amoebic keratitis. Resource water is a potential risk factor for the distribution of free-living amoeba. Therefore, more attention of health authorities to determine, training and prevention from infection are recommended.

Background: Leishmaniasis is important vector-borne parasitic disease worldwide, caused by the genus Leishmania. The objective of the current study was to identify genetic polymorphism in L. major, one of the species causing cutaneous leishmaniasis (CL), isolated from southeastern Iran, using Permissively Primed Intergenic Polymorphic-Polymerase Chain Reaction (PPIP-PCR) method. Methods: Overall, 340 patients with suspected CL were examined. They referred to the Central Laboratory in Chabahar, Iran during Apr 2013 to Feb 2014. Microscopic examination of Giemsa-stained slides from lesions as well as aspirates cultured in Novy- Mac Neal-Nicolle (NNN) Media was employed in order to diagnose CL in these patients. Our analyses detected 86 suspected subjects as having CL from which 35 isolates were cultured successfully. PPIP-PCR method was performed on extracted genomic DNA from selected isolates in order to determine the genetic polymorphism among L. major isolates. Results: The electrophoresis patterns demonstrated two genetic profiles including A or A1 patterns between all samples tested. Frequency of A and A1 sub-types were 33 (94.3%) and two (5.7%), respectively. Conclusion: Both host and parasite factors may contribute to the clinical profile of human leishmaniasis in the endemic foci of the disease. Here we showed that genetic variations pertaining to the Leishmania parasites might determine, in part, the clinical outcomes of human leishmaniasis.

Background Zoonotic cutaneous leishmaniasis (ZCL) is polymorphic disease that may show various clinical manifestations. Objectives This study investigates the determination of genetic variation within the species of Leishmania major isolates from new cases in Chabahar, a port city in Southeast Iran (situated at the Iran-Pakistan border). Migration in this region indicates that leishmaniasis is spreading gradually, and a new micro-habitat focus appears each year. Materials and methods A variety of nucleic acid detection methods that target both DNA and RNA have been developed. The restriction fragment length polymorphism analysis of amplified internal transcribed spacer 1 with polymerase chain reaction (ITS1-RFLP PCR) assay is a multipurpose tool for the diagnosis of Leishmania from clinical samples and for enabling the determination of the infecting Leishmania species. The goal of this study was the identification of species based on ITS1-RFLP in the ribosomal operon of L. major from clinically different forms of ZCL amplified by PCR, followed by the digestion of the PCR product with restriction enzymes. The profiles were observed and visualized in agarose gel under UV light. We used direct smears to identify the parasites. While taking the smear, samples were collected for culture or direct PCR. We used the PCR-RFLP assay of the ITS1 genes for direct identification of Leishmania species in 24 out of 33 suspected patients. PCR-ITS1 amplification was done on the 24 samples confirmed by culture via growth and parasitological methods. Results Of the 24 isolates, 21 had 350 bp bands (87.5%) and three had 450 bp bands (12.5%). After using the restriction enzyme, banding patterns including fragments of 210 and 140 bp for L. major were detected in 19 cases. Conclusions The L. major species causing ZCL in Chabahar have limited genetic variation. There seems to be little manifestation of diversity between these lesions as a new focus of disease, and new micro-habitats for the disease are appearing in parts of this region.

Background Hepatitis is a serious blood born infection in patients with β-thalassemia major (β-TM). There was no previous report on hepatitis prevalence in patients with β-TM in Zabol, Iran. Objectives The current study aimed to evaluate characteristics of hepatitis in patients with β-TM visiting Imam Khomeini Hospital of Zabol, in Sistan and Baluchistan province of Iran. Patients and Methods There were 90 patients with β-TM. General demographic and laboratory results were obtained by interviews and reviewing clinical and laboratory histories. Data for anti-HCV, HBs antigen and anti-HBs were extracted from latest archived records, and analyzed using SPSS ver. 19. Results Males and females comprised 51% and 49% of the patients, respectively. Mean age of the patients was 14.8±7.4 years old. Anti-HCV and HBs antigen were detected in 10% and 3.3% of the patients, respectively. Significant difference was found between the mean age of anti-HCV positive (20.1 ± 3.6, 95% CI: 17.3 - 22.9) and negative (14.2 ± 7.5, 95% CI: 12.5 - 15.9) patients (P = 0.02). Mean ferritin value was measured 4702 ± 2743 ng/mL. A significant correlation was observed between serum ferritin level and age (r = 0.3, P = 0.01). Hepatic enzymes, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) had mean levels of 51.8 ± 32.8 IU/L and 58.8 ± 59 IU/L, respectively. No significant association was identified between hepatitis status and either ferritin or hepatic enzymes levels. Conclusions Results demonstrated the relatively high prevalence of HCV infection in the patients. Applying sensitive methods to screen blood units is recommended to minimize the risk of transfusion associated hepatitis.

Background and Objective: Leishmania major is a flagellate protozoan parasite causing cutaneous leishmaniasis. Although pentavalent antimony compounds are the first-line drugs for leishmaniasis, their application is often accompanied by numerous limitations and side effects. Therefore, it is necessary to seek drugs of herbal origin that have fast-acting benefits and few side effects without resistance. This study aimed to evaluate the in vitro effects of methanolic extract of Arctiul lappa root on promastigotes and amastigotes of L. major. Methods: This experimental study evaluated the effects of 10, 100, 500, and 1000 µg/ml of A. lappa root methanol extract on L. major promastigotes using direct cell counting and MTT assay. The mean number of amastigotes in infected macrophages was calculated after 24 and 48 hours. Results: The half maximal inhibitory concentration (IC50) of A. lappa root methanolic extract was 131.25 µg/ml after 24 hours. The mean number of amastigotes in macrophages after 24 hours in the control group and in the A. lappa group with concentrations of 500 and 1000 µg/ml were 3.52, 2.02, and 1.27, respectively. Conclusion: The results show that the methanolic extract of A. lappa root has anti-leishmanial effects on the promastigotes and amastigotes of L. major in vitro. Keywords: Leishmania Major, Amastigotes, Promastigotes, Arctium.

Zoonotic cutaneous leishmaniasis (ZCL) is a polymorphic disease that may show various clinical manifestations. Although it is suggested that the genetic variability of the parasite is one of the factors influencing clinical manifestations of leishmaniasis, no data exists regarding genetic polymorphism of Leishmania major (L.major). This study investigates the determination of genetic variations within the species of L.major isolates from different cases of ZCL in two hyper-endemic areas of Iran.A variety of nucleic acid detection methods that target both DNA and RNA have been developed. Among these, the polymerase chain reaction (PCR) method proved to be a highly sensitive and specific technique. Species identification was based on permissively primed intergenic polymorphic-polymerase chain reaction (PPIP-PCR) and restriction fragment length polymorphism analysis of amplified internal transcribed spacer (ITS-RFLP) in the ribosomal operon of L.major from clinically different forms of ZCL. The DNA products were amplified by PCR, followed by digestion of the PCR product with restriction enzymes. The profiles were visualized in agarose gel under ultraviolet (UV) light.The PCR product obtained for all isolates was about 1060 bp in size. Different patterns of PPIP-PCR and ITS-RFLP in the ribosomal operon were classified as I, II, III, IV, and V. This classification was according to the number and localization of bands. Results of this research detected the genetic and clinical polymorphism of L. major, and showed that strain A was more frequent than other strains.The L.major causing ZCL in Isfahan, Iran is genetically a highly polymorphic species and PPIP-PCR exposed more genetic polymorphism among clinical samples in Isfahan, Iran.

Background: Ticks are important vectors of human and animal pathogens. They are considered as main vectors fortransmission of many viral, bacterial, rickettsial and parasitical pathogens. The aim of the present study was to find out species diversity of ticks, which infested the domestic ruminants in Zabol County, Eastern Iran in 2012. Methods: Ticks were selected randomly from sheep, goats, cattle and camels. The ticks were collected from the body of infested animals and stored in 70% ethanol, then transported to the laboratory of Zabol University of Medi- cal Sciences. Following examinations under stereomicroscope, ticks were identified using available taxonomic keys. Results: In this study, a total number of 469 adult ticks (381 males and 88 females) were collected. Ticks were clas- sified into 2 genera and 9 species including: Hyalomma dromedarii (17.3%), Hy. schulzei (1.8%), Hy. marginatum (0.5%), Hy. anatolicum excavatum (12.60%), Hy. anatolicum anatolicum (11.2%), Hy. asiaticum asiaticum (11.0%), Rhipicephalus sanguineus (21.2%), Rh. bursa (10.2%) and Rh. turacunis (13.911%). The frequency of genus Hyalomma (54.6%) was higher than Rhipicephalus. Rh. sanguineus was the predominant tick species and accounted for 21.26% of the ticks. The ratio of males was more than the female ticks. Conclusion: Hyalomma and Rhipicephalus spcies are commonly distributed in the study area. Further investiga- tions are needed to identify the role of above tick species as vectors of pathogenic organisms.