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Cystic Echinococcosis (CE) is a medically important disease that is caused by the metacestodes of Echinococcus granulosus. Human hydatid is considered an endemic disease in specific regions of Iran. The goal of the present study was to determine the genetic diversity of E. granulosus from the paraffin-embedded human tissue samples which were collected from the endemic regions of Iran. Fifty-five formalin-fixed and paraffin-embedded hydatid cysts (FFPE) of humans, which had been removed surgically, were obtained from the South Khorasan and Sistan and Baluchistan provinces. These regions are related to the East and Southeast regions of Iran, respectively. The cox1 and nad1 genes from mitochondria were amplified from the extracted DNA and sequenced. The sequences were edited using the BioEdit software. Furthermore, phylogenetic and genetic diversity analyses were performed. Sequencing of the cox1 and nad1 genes from the 44 CE samples was done successfully. Genetic analysis revealed that 38 (86.3%) and 6 (13.6%) of the isolates were G1- and G6-genotypes, respectively. In general, eight and six haplotypes were identified by cox1 and nad1 genes analysis, respectively. For G1 strains, the haplotype diversity index was higher for the cox1 gene (0.6 ± 0.07) in comparison with the nad1 gene (0.4 ± 0.09). The findings of the present study showed that the sheep strain (G1) and the less important camel strain (G6) play the main roles in the transmission cycle of CE in the East and Southeast regions of Iran. Therefore, these results could be useful for managing the hydatid disease control programs in the studied and other similar areas.

Malaria is one of the most serious health problems in many countries, including Iran. Accurate diagnosis is important regardless of the elimination status of a country. A cross-sectional study was performed on 105 people who were suspected to be positive for malaria infection in Sistan and Baluchistan, Iran. Blood smears (thin and thick films) were stained with 10% Giemsa. DNA was extracted from the prepared thin and thick films for molecular methods. Multiplex/nested polymerase chain reaction (mn-PCR), loop-mediated isothermal amplification (LAMP), and light microscopy (LM) were compared with nested PCR (nPCR) as a gold standard. Of 105 subjects, 52 (49.5%), 58 (55.2%), 58 (55.2%), and 63 (60%) were positive for malaria by LM, nPCR, mn-PCR, and LAMP, respectively. The sensitivity, specificity, and kappa were 92.1%, 100%, and 0.9 for LAMP and 100%, 100%, and 1 for mn-PCR, respectively. Eight cases of coinfection (Plasmodium vivax and Plasmodium falciparum) that were not detected by LM method were diagnosed by mn-PCR and LAMP. In the present study, the high sensitivity and specificity of LAMP and mn-PCR indicate that these two tests are good alternatives to nPCR for malaria diagnosis

Molecular analysis of antifolate resistance-associated genes—dihydrofolate reductase (dhfr) and dihydropteroate synthetase (dhps) of Plasmodium vivax is important in predicting the emergence of drug resistance to sulphadoxine-pyrimethamine (SP). The present study aimed to determine the polymorphism of dhfr and dhps genes in P. vivax field isolates. Samples from 80 microscopically diagnosed vivax malaria cases were collected from endemic areas of malaria in Hormozgan Province of Iran, from June 2010 to November 2015. The two sets of codons at position 33, 57, 58, 117, 173 of dhfr and 382, 383, and 553 of dhps genes were analysed by direct sequencing of PCR products. The majority of the isolates (70%) harboured a wild-type allele for P. vivax dhfr (Pvdhfr) and P. vivax dhps (Pvdhps). Mutations were detected in three codons of Pvdhfr (P33L, S58R and S117N) and single codon in Pvdhps (A383G). Novel mutations that have not been identified previously at codon 459 (D459A) of Pvdhps were also observed. The high prevalence of point mutation as well as the rising triple mutation of Pvdhfr and Pvdhps genotypes necessitate change in programmes and guidelines to eliminate P. vivax in future.

Background and objective Microsporidia are obligate intracellular pathogens. The parasite can infect a wide range of vertebrate and invertebrate hosts among all over the world. The current study was performed to evaluate the prevalence of intestinal microsporidiosis by staining and molecular methods in South Khorasan province of Iran. Materials and methods From December 2019 to December 2020, 720 stool samples were stained by specific Weber Modified Trichrome (Chromotrope 2R) to detect Microsporidia spores. The DNA extraction was done by Kit and To amplify the SSU rRNA gene of Microsporidia Spp, the nested PCR was used with genus specific primers. Finally data analyzing were performed by chi square (X2) test, using SPSS software. Results From 720 stool samples, 25 (3.47%) samples were microscopically positive for intestinal Microsporidia infection. Out of 25 positive samples of Microsporidia in microscopic method, 22 samples were positive in PCR method. In this study, most of the molecularly approved patients were referred to the hospital (68.00%) and were in the age range of 30 to 50 years (40.80%). 92.00% of patients also had gastrointestinal symptoms and were infected in the summer (40.00%), (p Conclusions This study was the first evaluation of the prevalence of Microsporidia in South Khorasan province. Microsporidia infection is prevalent in the public population of eastern Iran. Statistical analysis of the results showed that factors such as education, age, gastrointestinal symptoms and season can be risk factors associated with parasitic infections.

Toxoplasma gondii is zoonotic protozoa that spread throughout the world. The aim of current study was determination of the prevalence of T. gondii infection in patient undergoing chemotherapy for malignancies using ELISA and two different molecular tests including Nested-PCR and LAMP techniques. This study was carried out in Ali Ibn Aboutaleb hospital in Zahedan cities, southeast of Iran. Totally, 154 serum samples from patient were test for detection of IgG and IgM antibodies against T. gondii by ELISA test. All of the samples were also test by Loop amplification (LAMP) and Nested-PCR with RE gene. Totally, 39.4% of the sample sera from cases were positive for IgG against T. gondii. After using RE-Nested-PCR technique, 29.8% sample sera were positively. After using LAMP technique, 33.1% of the samples were found to be positive. Based on our result, the RE-LAMP technique was more effective than RE-nested-PCR method in the detection of T. gondii DNA.

Cutaneous leishmaniasis (CL) is one of the most common parasitical diseases in many regions of Iran and plays a significant role in the inhibition of socioeconomic growth of this deprived province. The present study aimed to molecular identification of cutaneous leishmaniasis species in patients from Chabahar and Konark district, in Sistan and Baluchestan province, Iran. This descriptive cross-sectional study was carried out on 250 patients suspected of cutaneous leishmaniasis (CL). Samples of the skin lesions and ulcers suspected to CL were collected from patients and diagnosed by parasitological test and molecular methods, respectively. The microscopic test detected 80 out of 250 (32%) suspected patients to CL, while 80 out of 80 (100%) were positive for Leishmania kDNA. Two species L. major and L. tropica were responsible for 9 and 69 cases, respectively. Our results were consistent with the previous reports in Iran, revealing L. tropica as the most causative agent of CL Further investigation are demanded to identify the hosts, reservoirs, disease species and vectors in this part of the world.

Dirofilaria immitis is endemic in the southern parts of Iran where there is a favourable environment for the development of Anopheles stephensi. The aim of the present study was to evaluate the susceptibility of A. stephensi as D. immitis vector under laboratory conditions. Anticoagulated dog blood containing 1,500 microfilariae ml–1 was artificially administered to 140 mosquitoes of the test group (T group), while 94 mosquitoes were left as a control (C group). Blood-fed mosquitoes of the T group were dissected for morphological and molecular analyses at pre-set time points to observe the developmental stage of D. immitis. The results showed that the average number of the microfilariae ingested per female in T group was 9.6. At the end of the study, 16 mosquitoes developed infective thirdstage larvae of D. immitis, and 49 mosquitoes survived at end of incubation period, giving an estimated infection rate of 32.6% and the vector efficiency index of 17%. The infection rate and vector efficiency index suggested that A. stephensi could be considered as a potential vector of D. immitis., Нематоды Dirofilaria immitis обычны в южных провинциях Ирана, где складываются походящие условия для развития комаров Anopheles stephensi. Проведено исследование способности A. stephensi служить вектором-переносчиком для нематод D. immitis в лабораторных условиях. Кровь собак с добавлением антикоагулянтов, содержащие 1,500 микрофилярий в 1 мл была использвоана для заражения голодавших самок комаров. Всего 140 комаров было использовано для эксперимента (Т группа) и 94 особи комаров служили контролем (C группа). Комаров из Т группы вскрывали через определенные промежутки времени для морфологического и молекулярного выявления развивающихся стадий D. immitis. Результаты показали, что в среднем каждая самка комара заглатывала 9.6 микрофилярий. В конце эксперимента было установлено, что 49 самок пережили условия данного опыта (32.6%) и у 16 самок комаров развились инвазионные личинки 3-й стадии D. immitis. Таким образом, индекс эффективности переноса составил 17%. Сделан вывод о способности A. stephensi служить вектором для D. immitis.

We report a case of urinary myiasis occurring in a 60-yr-old Iranian male patient with urinary tract problems and a history of travel to Thailand who was referred to Shafagh Medical Laboratory in Tehran (Iran). Larvae excreted in the patient's urine were confirmed by morphological identification key and DNA barcoding to belong to the species Megaselia scalaris Loew, which is known as the scuttle fly. Based on the patient's history, he was infected with M. scalaris in Thailand. To the best of our knowledge, this is the first report of urinary myiasis caused by M. scalaris in Thailand.

This study was conducted to address the distribution of Acanthamoeba genotypes in therapeutic hot springs in Iran. Sixty water and sediment samples were collected from bicarbonate, sulphur, and sodium chloride thermal springs in the northwest. All hot springs examined are used mainly for health purposes in Iran. Acanthamoeba were identified by both morphology and PCR (polymerase chain reaction). Genotype identification was based on the sequencing of a highly variable and informative region of Diagnostic Fragment 3 (stem 29–1 of 18S rRNA gene) within Acanthamoeba-specific amplimer (ASA.S1). Twenty percent of hot springs were contaminated with thermotolerant Acanthamoeba belonging to the potentially pathogenic T4 and T3 genotypes. A high number (91.7%) of strains showed growth at 37 °C, and eight isolates showed growth at 42 °C. A single isolate (HSNW2) was detected in waters at 70 °C. The presence of thermotolerant Acanthamoeba highlights a risk factor for susceptible individuals, as Acanthamoeba-related keratitis continues to rise in Iran. Periodic surveillance of thermal waters as well as improved filtration and disinfection is recommended to prevent disease related to pathogenic Acanthamoeba. This is the first comprehensive molecular study of Acanthamoeba genotypes in hot springs in Iran and the first to report the occurrence of the T3 genotype (corresponding to Acanthamoeba griffini) in thermal water sources in this country.