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Objectives: This research aimed to investigate the impact of ozone on Leishmania major.Methods: Leishmania major promastigotes were exposed to ozone flow at varying concentrations (5, 10, 20, 30, and 40 ppm) for different durations (5, 10, 20, 30, and 40 minutes). The DNA of the treated promastigotes was extracted to evaluate DNA fragmentation. Apoptosis induction in the treated L. major promastigotes was analyzed using a specific kit. Additionally, scanning electron microscopy was employed to observe ultrastructural changes in the treated promastigotes.Results: The highest fatality rate (100%) occurred after 30 minutes of exposure to 40 ppm ozone gas, while the lowest fatality rate was 16.14% after 5 minutes of exposure to 5 ppm ozone gas. The DNA of the treated parasites was degraded, and significant morphological changes were observed in the body and flagella of the promastigotes. Induction of apoptosis was also noted.Conclusion: Ozone exhibits anti-Leishmania activity and induces apoptosis in Leishmania major. Furthermore, clear ultrastructural changes were observed in Leishmania major following ozone exposure. These findings suggest that ozone warrants further investigation as an anti-leishmanial agent.

Aim: Despite the various studies done in the field of cutaneous leishmaniasis treatment, there is still no perfect and safe drug for definite treatment of this disease. Therefore, the quest to find an appropriate drug continues. Previous studies have shown that auranofin has anti-leishmanial activity; therefore, in this study; the effect of auranofin on Leishmania major was studied. Materials and Methods: Effectiveness of four concentrations (1, 2, 4, and 8 μg/ml) of auranofin on L. major was studied. After culturing amastigotes and promastigotes of the parasite, IC50 was calculated. The ability of auranofin to induce apoptosis in promastigotes was evaluated, and the degree of fragmentation of promastigotes DNA after treatment with auranofin was studied. Subsequently, the ultrastructural changes induced by treatment with auranofin in promastigotes were studied. Using the obtained results, IC50 of auranofin against amastigotes and promastigotes was calculated as 1.007 and 2.38 μg/ml, respectively. Findings: Showed that auranofin induce apoptosis in L. major. The highest rate of apoptosis (%80.1) occurred at the concentration of 8 μg/ml and also auranofin-induced fragmentation of DNA. Considerable changes occurred in the shape of body and free flagellum of the L. major promastigotes after treatment with auranofin. Conclusion: Based on the results, it can be concluded that auranofin has a considerable anti-leishmanial activity and additional studies in this field will be based on the results.

Aim: Cattle is one of the main sources of food supply chain for humans in most countries. The present study aimed to identify the infection rates for Toxoplasma gondii, Neospora caninum, and Sarcocystis spp. by polymerase chain reaction (PCR) method in native slaughtered cattle in Kashan, central Iran. Materials and Methods: Totally, 159 diaphragm, esophagus, and muscle samples (53 samples of each) were collected from native beef cattle from Kashan slaughterhouse, central Iran. The genomic DNA was extracted, and PCR method was used separately for detection of N. caninum, Sarcocystis, and T. gondii species using specific primers. Finding: Sarcocystis was found in 84.9% of muscles, 83% of esophagus, and 84.9% of diaphragm samples. Mixed infection (Sarcocystis cruzi–Sarcocystis hominis) was the most common infection, followed by S. cruzi and S. hominis. Sarcocystis hirsuta was not detected in any samples. T. gondii was detected only in three (5.7%) out of the 53 muscle tissues samples of cattle. N. caninum was found in 18.9% of muscles, 24.5% of esophagus, and 28.3% of diaphragm samples. One of the cattle had coinfection to Neospora, Toxoplasma, and Sarcocystis in muscles simultaneously. There was no statistically significant difference between infection rates and age as well as sex in each organ. Conclusion: This study revealed a low prevalence rate of T. gondii, but a high prevalence of infection to N. caninum and S. cruzi or mixed infection of S. cruzi with S. hominis among slaughtered cattle. Prevention measures such as keeping away dogs from cattle grazing are recommended.

Aims: The control of fascioliasis has depended on the utilization of a predetermined number of anthelmintic drugs. Nonetheless, the resistance of antifasciolid is presently boundless, and there is a need for potential drug properties of medicinal plants as new medications against Fasciola spp. to interrupt the parasite transmission. Materials and Methods: This investigation is meant to assess the potential anti-fasciolicide impacts of Zingiber officinale roscoe hydroalcoholic extract against the Fasciola miracidia. The eggs of Fasciola hepatica were gathered from the livers contaminated goats and sheep and washed 20 times with chlorine-free water, and afterward incubated at different pH, temperature, and light density for embryonic development. Concentrated miracidia of Fasciola spp. were incubated at different concentrations (2, 5, and 10 μg/mL) of hydroalcoholic extract of Z. officinale at different time intervals. The data were analyzed using the ANOVA statistical test. Results: The extract at different concentrations showed antifasciolid effects in comparison to triclabendazole. This braking action was dose-proportional and further related firmly to the disclosure time ( P < 0.001). In concentration of 10ug/ml extract killed the parasites at 105 ± 3 s ( P < 0.001). Conclusion: In vitro antimiracidia activities of the hydro alcoholic extract of Z. officinale was satisfactory and potent fasciolicide effective, however, in vivo impact of this extract, remains for extra assessment. In this manner, these therapeutic plant extracts might be seen as confident origins of bioactive composites that could be matured against miracidia. This is the main work to evaluate the potential enemy of fasciolicide impact of certain plants against Fasciola miracidia.

Aims: Hydatidosis is an important zoonotic disease that is caused by a tiny tapeworm, namely Echinococcus granulosus. In this study, three polymerase chain reaction (PCR)-based methods, including, high resolution melting (HRM) analysis, DNA sequencing, and PCR-restriction fragment length polymorphism (RFLP) have been used for genotype the identification of E. granulosus isolates from dogs and camels in Zarinshahr and Najafabad, Isfahan province, Iran. Materials and Methods: A total of 200 adult worms of 40 dogs and 51 samples of camel hydatid cysts were examined. Molecular characterization of isolates was performed using HRM assay, sequencing of DNA, and digestion Rsa1 pattern coding for the mitochondrial cox1 gene. For analysis of the HRM melting curve, we used the Tm within the range of 77.50°C–79.23°C. Results: HRM analysis revealed that 72.5%, 15%, and 12.5% dog's genotypes and 41.17%, 21.56%, and 35.29% camel genotypes were G1, G3, and G6, respectively. PCR-RFLP analysis, spare parts 310 bp and 138 bp of cox1 that shows the G1 genotype in all of the isolates. Sequence analysis as well as HRM assay was confirmed genotypes of G1, G3, and G6 in camels and dogs. Based on three methods of the cox1 gene the dominant genotype was G1. Conclusion: The PCR-RFLP only identified the G1 genotype, whereas the HRM analysis, as well as DNA sequencing, were detected three genotypes G1, G3, G6, therefore, these two methods have enough accuracy for the determination of genotypes of E. granulosus. This information leads to a better understanding of the biological characteristics of E. granulosus genotypes in Iran and shows the camel as a source of human hydatidosis.

Aims: Blastocystis species are one of the most common enteric protist infections in humans and some animals worldwide. Molecular studies have shown that there is a high level of genetic variation among Blastocystis isolates. The aim of this study was to identify the subtypes and frequency of Blastocystis isolates in patients who referred to the medical diagnostic laboratories in Kashan, Central Iran. Materials and Methods: This cross-sectional study was conducted on 1118 patients, from December 2017 to June 2018. Fecal specimens were evaluated by the microscopic examination. Positive samples were cultivated in Robinson media. After massive growth and DNA extraction, a 550 bp from the small subunit ribosomal RNA gene was amplified by the polymerase chain reaction (PCR) for subtype identification. The PCR products have been sequenced, identified, and compared at the NCBI site. The results were analyzed using the SPSS software version 16. Results: The frequency of Blastocystis sp. was 8.58%, (confidence interval = 6.94%–10.22%) from which 76% were men and 24% were women. Of the 51 PCR positive samples, ST3 (41.2%), ST1 (39.2%), ST2 (11.8%), and 7.8% isolates were identified as mixed. ST3 and ST1 have been more common. The highest levels of infection were observed in the food-handlers, the age group of 31–40, and people with high school education. Conclusion: The results showed that the frequency of Blastocystis was lower than other studies and the most common Blastocystis subtype was subtype 3, followed by subtype 1, and subtype 2.

Aims: In vitro cultivation of digenea would help the development of effective treatments and studies of the biology of the parasites. The goal of the present study was to optimize culture conditions for the maintenance liver fluke, Dicrocoelium dendriticum. Materials and Methods: Forty fresh D. dendriticum were collected from the sheep liver and washed three times with warm Roswell Park Memorial Institute (RPMI) 1640 Medium. The collected worms were transferred to 24-well Nunc-Immuno plates containing RPMI media supplemented with 50% of fetal bovine serum (FBS), 2% of sheep red blood cells (RBCs), 50 IU/ml of penicillin, and 50 mg/ml streptomycin. The mobility of the live/dead worms was observed by inverted microscope. The mean and median survival time was calculated by Kaplan–Meier model, and survival and hazard function graphs were also analyzed. Results: D. dendriticum was lived in vitro only for long periods of about 25 dyes. The 1st day of maintaining in culture media, one worm was dead and the number of dead worms was raised to 40 after 25 days of incubation. On the one hand, the mean survival time was 392 h with a confidence interval (CI) of 95% (384.8–400.03). On the other hand, the median survival time was 420 h with a CI of 95% (406.9–433.09). D. dendriticum was able to be alive in RPMI 1640 media for at least 25 days. Conclusion: RPMI 1640 supplemented with FBS, and RBCs can be used as short-term maintenance for the in vitro culture of D. dendriticum. The outcomes of the current study could be useful for many aspects of parasitological analysis.