Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 294 300 Seyed AliAkbar Shamsian Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran Hamed Mirizadeh Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran Ghodratollah Salehi Sangani 1. Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran 2. Cutaneous Leishmania Research Center, Mashhad University of Medical Sciences, Iran Mahdi Zarean 1. Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran 2. Cutaneous Leishmania Research Center, Mashhad University of Medical Sciences, Iran Fariba Berenji Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran Mohsen Najjari Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran Abdolmajid Fata 1. Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran 2. Cutaneous Leishmania Research Center, Mashhad University of Medical Sciences, Iran Elham Serki Department of Virology & Microbiology Research Center, Mashhad University of Medical Science, Mashhad, Iran Vahideh Tahreian Laboratory of Molecular Reference and Tuberculosis in the Northeast of Iran, Dr. Ali Shariati Hospital, Mash-had University of Medical Science, Iran Bibi Razieh Hoseini Farash Department of Parasitology and Mycology School of Medicine, Mashhad University of Medical Science, Iran 2. Cutaneous Leishmania Research Center, Mashhad University of Medical Sciences, Iran 2022 11 13 2023 04 10 Background: Epidemiological studies, classification and genetic studies of Leishmania species are effective in treatment, control and prevention in endemic areas. We aimed to investigate the genetic diversity and phylogeny of Leishmania in Zoonotic foci located in northeastern Iran using nagt gene for the first time. Method: DNA of 100 confirmed positive slides collected from the health centers of Sarkhes, Darghez, Fariman, Esfarayen, and Sabzevar were extracted during 2020-2021. The partial sequence of kDNA was amplified to identify the species. Twenty-five DNA samples were randomly subjected to amplify by nagt gene primes and were sequenced. The sequences were aligned with reference sequences in National Center for Biotechnology Information (NCBI). Then, the genetic similarities of the sequences were checked using Clustalx2.1 software and the phylogenetic tree was drawn by Mega 7 software. Result: All the positive samples were diagnosed as L. major. Approximately, half of the sequences of species were similar to two reference genes JX103550.1:404-712 L. major Esfahan and KX759012.1:568-807 L. Major Ilam (more than 90% similarity). According to the results of the phylogeny tree, the closest genotype to our study samples was JX103550.1:404-712 L. major Esfahan. Conclusion: The most causative agent CL in these areas was L. major. The genetic diversity of L. major was high such as other zoonotic foci in Iran. Due to the high similarity of the strains in the study areas with the strains of Isfahan and Ilam, similar control and prevention methods is suggested in these areas.  https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3789 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3789/1301

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 279 293 EN Mehdi Mohebali 1. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2. Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran Gholamhossein Edrissian Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Behnaz Akhoundi Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Mohammedreza Shirzadi 1. Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran 2. Center for Communicable Diseases Management, Ministry of Health Treatment and Medical Education, Tehran, Iran Gholamreza Hassanpour Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran Atefeh Behkar Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran Yavar Rassi Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Homa Hajjaran Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Hossein Keshavarz 1. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2. Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran Mohammad Mehdi Gouya Center for Communicable Diseases Management, Ministry of Health Treatment and Medical Education, Tehran, Iran Shahnam Arshi Center for Communicable Diseases Management, Ministry of Health Treatment and Medical Education, Tehran, Iran Mohammad Zeinali Center for Communicable Diseases Management, Ministry of Health Treatment and Medical Education, Tehran, Iran Zabihollah Zarei Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2. Meshkin-Shahr Health Station from the School of Public Health, Tehran University of Medical Sciences, Ardebil, Iran Iraj Sharifi Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran Zahra Kakooei Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2023 06 07 2023 07 25 Background: Visceral leishmaniasis (VL) is one of the most important neglected tropical diseases. The zoonotic form of VL is endemic in some areas of Iran. We aimed to determine the status of VL identified in humans and canines in different parts of Iran from 2013 to 2022. Method: A national representative cross-sectional study was conducted in 10 provinces of Iran, including the national leishmaniasis reference lab. We employed the direct agglutination test (DAT) as a reliable serological method to detect anti-Leishmania infantum antibodies in humans and animal reservoir hosts. Additionally, a narrative literature review was conducted to identify relevant studies on VL seroprevalence in Iran from 2013 to 2023. Results: The results of 21281 human and 5610 canine serum samples from 2013 to 2022 are reported. Altogether, 448 (2.1%, 95%CI: 2.0-2.3) human serum samples showed anti-L. infantum antibody levels of ≥1:3200. Of these samples, 13716 (64.5%) were collected actively, which showed a seroprevalence of 0.6% (95% CI: 0.5-0.8) and 7565 (35.5%) were collected passively, which showed a seroprevalence of 4.8% (95%CI: 4.3-5.3). Overall, 1035 (20.1%, 95%CI: 19.0-21.2) of 5160 domestic dogs (Canis familiaris) samples showed anti-L. infantum antibody levels of ≥1:320. Northwest (2.8%) and northeast (0.96%) regions had the highest human VL seroprevalence, while northwest (21.5%) and south (14.4%) regions had the highest canine VL seroprevalence. Conclusion: Zoonotic VL, an endemic parasitic disease, is still present in several different distinct areas across Iran. While human VL cases have shown a declining trend over the last decade, the prevalence of canine VL remains significant. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3973 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3973/1300

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 400 403 Taliha Karakök Infectious Disease Deparment, Fatsa State Hospital, Ordu, Türkiye 2022 11 16 2023 09 22 Splenic infarction is a rare complication of both malaria and COVID-19. We report a splenic infarction case due to COVID-19 and malaria co-infection. A 35‑year‑old male with no known chronical disease tested positive for both COVID-19 and malaria in Turkey in 2022. Oral artemether and lumefantrine treatment was started. On the third day of the treatment, he complained about a severe left upper quadrant pain. A repeated abdominal CT showed splenomegaly and 8 cm diameter hypodense areas throughout the spleen consistent with splenic infarction. The patient was discharged with low molecular weight heparin. A rare complication that can be seen in both diseases developed a more rigorous recommendation for anticoagulant therapy is needed for co-infections of COVID-19 with diseases that may present similar thrombotic complications. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3797 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3797/1313

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 301 312 Zahra Hosseininejad Toxoplasmosis Research Center, Communicable Disease Institute, Mazandaran University of Medical Sciences, Sari, Iran 2. Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran Ahmad Daryani Toxoplasmosis Research Center, Communicable Disease Institute, Mazandaran University of Medical Sciences, Sari, Iran Mahdi Fasihi-Ramandi Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Hossein Asgarian-Omran Department of Immunology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran a Reza Valadan Department of Immunology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Tooran Nayeri Toxoplasmosis Research Center, Communicable Disease Institute, Mazandaran University of Medical Sciences, Sari, Iran 2. Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran Samira Dodangeh Department of Medical Parasitology and Mycology, Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran Shahabeddin Sarvi Toxoplasmosis Research Center, Communicable Disease Institute, Mazandaran University of Medical Sciences, Sari, Iran 2022 06 20 2023 09 22 Background: We aimed to design a B and T cell recombinant protein vaccine of Toxoplasma gondii with in silico approach. MIC13 plays an important role in spreading the parasite in the host body. GRA1 causes the persistence of the parasite in the parasitophorous vacuole. SAG1 plays a role in host-cell adhesion and cell invasion. Methods: Amino acid positions 73-272 from MIC13, 71-190 from GRA1, and 101-300 from SAG1 were selected and joined with linker A(EAAAK)A. The structures, antigenicity, allergenicity, physicochemical properties, as well as codon optimization and mRNA structure of this recombinant protein called MGS1, were predicted using bioinformatics servers. The designed structure was synthesized and then cloned in pET28a (+) plasmid and transformed into Escherichia coli BL21. Results: The number of amino acids in this antigen was 555, and its antigenicity was estimated to be 0.6340. SDS-PAGE and Western blotting confirmed gene expression and successful production of the protein with a molecular weight of 59.56kDa. This protein will be used in our future studies as an anti-Toxoplasma vaccine candidate in animal models Conclusion: In silico methods are efficient for understanding information about proteins, selecting immunogenic epitopes, and finally producing recombinant proteins, as well as reducing the time and cost of vaccine design. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3656 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3656/1302

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 404 407 Anil Kumar Department of Veterinary Medicine, Bihar Veterinary College, BASU, Patna, Bihar, India Pankaj Kumar Department of Veterinary Parasitology, Bihar Veterinary College, BASU, Patna, Bihar, India Sonam Bhatt Department of Veterinary Medicine, Bihar Veterinary College, BASU, Patna, Bihar, India Ajeet Kumar Department of Veterinary Biochemistry, Bihar Veterinary College, BASU, Patna, Bihar, India G.D Singh Department of VCC, Bihar Veterinary College, BASU, Patna, Bihar, India Ankesh Kumar Department of VCC, Bihar Veterinary College, BASU, Patna, Bihar, India 2022 12 28 2023 04 10 Bovine tropical theileriosis is one of the potentially fatal disease of dairy cattle, which is caused by hemoparasite Theleria annulata. About seven years old cross-bred cow was presented with complaint of pyrexia, inappetance, lacrimation and ocular swelling since last 5 days. The clinical examination showed elevated rectal temperature (39.4 0C), mild enlarged pre-scapular lymph nodes, bilateral bulging of temporal fossa, protruded pale and icteric conjunctivae of the eyes with lacrimation and presence of ticks on body. The case was suspected for haemoprotozoan disease. Blood and serum sample were collected for hematological, blood smear examination and molecular examination (PCR), and biochemical analysis respectively. Microscopic examination of blood smear revealed intra-erythrocytic signet ring shaped periplasm of Theileria annulata. Hemato-biochemical examination revealed anemia, hypoproteinemia, hypoalbuminemia and jaundice. Further, PCR assay was done using T. annulata-specific primer pair, Cyto b1 gene targeting the amplicon of 312 bp showed specific band on Gel-electrophoresis. Therapeutic regimen was started with Buparvaquone @ 2.5 mg/kg body weight IM single dose followed by Oxytetracycline @ 10 mg/kg body weight IV in 500 ml of NS for 5 days and Prednisolone @ 0.25 mg/kg body weight IM for 3 days along with supportive therapy. The cattle well responded to the therapy and complete regression of ocular signs was observed within one week of treatment. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3843 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3843/1314

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 313 323 Mehran Mirzaghavami Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Javid Sadraei Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Majid Pirestani Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Saeed Bahadory Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2022 09 26 2023 09 22 Background:  We aimed to characterize Cryptosporidium spp. in rats, cats, pigeons, and crows. Methods: Fifty-five animal origin Cryptosporidium spp. genome were identified, genotyped and confirmed by nested PCR and of RFLP-PCR analysis as well as sequenced based on 18s rRNA and gp60 genes in Tehran (2012-2019). Finally, the phylogenetic analysis was performed by MEGA software (version 7). Results: By the molecular method, Cryptosporidium spp. were detected in 24 (15.2%), 15 (15%), 2 (2%) and 13 (13%) cases of wild rats, cat, pigeon, and crow, respectively. Among the identified species by the RFLP pattern, most isolates were identified as C. parvum (24/157) 17.8% in rats, (15/100) 15% in cats, (13/100) 13%in crew and (2/100) 2% in pigeons; and the rest of the cases were C. muris and C. felis. The results of sequencing did not prove the existence of C. parvum, C. felis, C. muris, and rat genotype. Subtyping of C. parvum was indicated that the dominant subtype family belongs to the IId family and the subtype A20G1 was the most common subtype detected in all hosts while A19G1 was detected in one isolate of cat and pigeon. Conclusion: Free-ranging animals are infected by species/subtype of Cryptosporidium, which can infect humans. This shows by itself the hygienic importance of the free-ranging animals in urban ecosystems. In the transmission of human cryptosporidiosis, the multi-host Cryptosporidium species such as C. parvum, C. felis, and C. muris can be transferred potentially from these animals to humans. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3604 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3604/1304

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 408 413 Noor Nihad Baqer Water and Environment Directorate, Ministry of Science and Technology, Baghdad, Iraq Ahmed Mohammed AL-Salam Hospital, Ministry of Health, Mosul, Iraq Bassad AL-Aboody Department of Biology, College of Science, University of Thi-Qar, Nasiriyah, Iraq Abdullah Ismail Public Health Laboratory, Ministry of Health, Mosul, Iraq 2022 09 23 2022 12 26 We diagnosed a case report of amoebic meningoencephalitis by Naegleria fowleri. This case represented the first recording in Iraq where it was not recording previously. This case was diagnosed after the death of an 18-year-old girl patient who lived in a rural area of Mosul in Iraq. Genetics detection of N. fowleri showed PCR product was 183bp for 18S rRNA gene. It was registered as the first recording of Iraqi isolate N. fowleri in GenBank with accession number OP380864.1. It is necessary to examine microscopically the cerebral spinal fluid (CSF) to observe the amoeba stages and exclude the bacterial causative. Rapid diagnosis may help in the treatment of amoebic meningoencephalitis. In addition, genetic identification can diagnose amoeba. Avoiding swimming or using freshwater contributes to prevent amoebic meningoencephalitis infection. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3741 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3741/1315

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 324 330 Fahimeh Kooshki Department of Pediatric Dentistry, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran Amal Khudair Khalaf Department of Microbiology, College of Medicine, University of Thi-Qar, Nasiriyah, Iraq Hossein Mahmoudvand Hepatitis Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran Parastoo Baharvand Hepatitis Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran Faezeh Gandomi Rouzbahani Student Research Committee, Lorestan University of Medical Sciences, Khorramabad, Iran Behnoush Selahbarzin 1. Hepatitis Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran 2. Department of Pediatric Dentistry, School of Dentistry, Lorestan University of Medical Sciences, Khorramabad, Iran 2022 06 04 2023 01 22 Background: This survey was designed to study the molecular epidemiology and risk factors of Entamoeba gingivalis and Trichomonas tenax in children with underlying malignancies and those on chemotherapy in Lorestan province, West of Iran. Methods: The present cross-sectional descriptive study was performed on children who suffering from different types of malignancies or receiving treatment by chemotherapy referring to oncology section of hospitals of Lorestan Province, Iran during May 2021 to April 2022. The frequency of oral cavity protozoa was investigated using microscopic and conventional polymerase chain reaction (PCR). Results: E. gingivalis and T. tenax parasites were found in 23 (25.5%) by microscopic method and 28 (31.1%) using PCR in children with malignancy. Among positive samples, 20 (71.4%) were infected with E. gingivalis; whereas 8 (28.6%) of the participants were positive for T. tenax. In the multivariate model, living in rural regions (OR= 3.437; 95% CI= 1.22-9.63; p=0.019) and using mouthwash (OR= 0.082; 95% CI= 0.018-0.37; p<0.001) were significantly related with the frequency of oral cavity parasites. Conclusion: Our results showed the high frequency of oral cavity parasites in children who suffering malignancies or receiving treatment by chemotherapy in Lorestan province, Iran. The awareness of the main risk factors for oral cavity parasites particularly using mouthwash is necessary in improving public and oral health strategies in children with cancer. Consequently, oncologist and dental practitioners must be aware to identify and manage oral health concerns in in children who suffering from different types of malignancies to prevent the oral diseases and infections. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3640 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3640/1305

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 414 418 Gökhan Pösteki Kocaeli State Hospital, Kocaeli, Turkey 2022 07 13 2023 09 22 Gallbladder is a rare localization for hydatid disease. Complications are even rarer and precise diagnosis is quite difficult even with radiological assistance. We report a rare case of 41-yr-old male patient presenting with the rupture of a gallbladder hydatid cyst with multiple fistulae to intra and extrahepatic bile ducts and pancreas, at Kocaeli State Hospital, Turkey in 2021. The patient had abdominal pain and abdominal CT scan reported a bizarre “contrast enhanced cholangiography” sign - radiopaque contrast substance in gallbladder, intra and extrahepatic bile ducts and pancreatic ducts, with concomitant acute cholecystitis. Surgery was performed and intraoperatively gallbladder hydatid cyst with multiple fistulae was noted. Cholecystectomy with total cyst excision was performed. Endoscopic retrograde cholangio pancreatography (ERCP) was utilized to irrigate and eradicate the parasite in the fistulae tracts localized near pancreas and intra/extrahepatic ducts. Postoperative period was uneventful, antiparasitic treatment was started, and in the yearly follow-up patient had no recurrence. Multidisciplinary and minimal invasive management is crucial in such bizarre, complicated cases. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3674 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3674/1316

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 331 341 EN Mahyar Khorram Department of New Sciences and Technologies, South Tehran Branch, Islamic Azad University, Tehran, Iran Heidar Masjedi Chronic Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran Fatemeh Tabrizi Department of Parasitology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Mitra Sadat Rezaee 1. Genomic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2. Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran Payam Tabarsi Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran Majid Marjani Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran Mihan Pourabdoullah Toutkaboni Chronic Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran Fatemeh Sheikholeslami 1. Pediatric Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Disease (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran 2. Department of Molecular Biology, Dr. Khosroshahi’s Pathobiology Laboratory, Tehran, Iran 2022 10 24 2023 08 12 Background: We aimed to evaluate the accuracy of genotyping of Leishmania species by the spliced leader mini-exon gene. Methods: Suspected leishmaniasis patients, referred to Masieh Daneshvary Hospital, Tehran, Iran were included from May 2017 to September 2021. The Leishmania species were genotyped by PCR-RFLP based on the SL mini-exon gene and the ITS1 region of SSU-rRNA gene and compared with the sequencing results. The expressed metabolites of metacyclic promastigotes were evaluated by Proton nuclear magnetic resonance (1H-NMR). Results: Out of 66 suspected cases, 36 (54.4%) were positive for Leishmania species based on the PCR assays. In 21 (31.8%) cases, promastigotes grew on culture tubes. Based on the RFLP of SL RNA profile, 13 (19.7%) L. tropica, 9 (13.6%) L. major, 3 (4.5%) L. infantum, and 8 (12.1%) C. fasciculata isolates, isolated from culture media, were identified; however, 3 (4.5%) cases were unidentifiable due to the low number of parasites. Seventeen metabolites were expressed by the metacyclic forms of L. major, L. tropica and C. fasciculata isolates. The top differential metabolites expressed more in C. fasciculata were FAD, p-Methoxybenzyl alcohol and S-b-G-5, 5-G-b-S (A = CH2) (P<0.005) whereas Veratryl glycerols and D-(+)-Mannose were significantly increased in L. major and Betulin, L-Tyrosine in L. tropica (P<0.01). Conclusion: The invaluable techniques such as sequencing and 1H-NMR confirmed the results of genotyping of Leishmania species based on the SL mini-exon gene.  SL mini exon gene can be used as a diagnostic tool to differentiate various Leishmania genotypes and detect contamination of culture media with C. fasciculata.  https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3765 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3765/1306

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 342 350 Muhammad Mustufa National Institute of Health, Health Research Institute, Islamabad, Pakistan Safia Bibi National Institute of Health, Health Research Institute, Research Centre, National Institute of Child Health, Karachi, Pakistan Syed Raza National Institute of Child Health, Karachi, Pakistan Syed Hasan National Institute of Child Health, Karachi, Pakistan Taranum Siddiqui National Institute of Health, Health Research Institute, Research Centre, Jinnah Post Graduate Medical Center, Karachi, Pakistan Allah Rakhia National Institute of Health, Health Research Institute, Research Centre, National Institute of Child Health, Karachi, Pakistan Muhammad Munir National Institute of Health, Health Research Institute, Islamabad, Pakistan Ayesha Merchant National Institute of Child Health, Karachi, Pakistan Waquarudin Ahmed National Institute of Health, Health Research Institute, Research Centre, Jinnah Post Graduate Medical Center, Ka-rachi, Pakistan 2022 08 11 2023 01 22 Background: More than 250 million people are infected by malaria parasites annually while around one million children less than 5 years of age die every year due to malaria. We aimed to assess the seasonal trends and usefulness of capillary and venous blood for rapid diagnosis of malaria. Methods: This cross-sectional study of 18 months duration was conducted at the National Institute of Child Health (NICH), Karachi. All patients reporting fever as chief complaint were recruited as study subjects. A semi-structured questionnaire was used to collect demographic information, presenting complaints, awareness of caregivers regarding malaria, preventive measures and history of malaria fever. Three ml Venous (2-3ml) as well as peripheral blood (3-4 drops) samples of all patients were collected for microscopy and rapid diagnostic tests (RDTs). Results: Out of total 477 patients with fever Venous and Capillary Blood RDTs methods detected 33(6.9%) and 30(6.3%) as the malaria positive while Venous and Capillary Blood Microscopy detected 30(6.1%) and 32(6.7%) cases respectively. Plasmodium Vivax infection was the most prevalent (87.9%) and majority (39.39%) of the cases occurred in the quarter, July to September. Conclusion: July to September is the peak season for malaria and P. Vivax (87.9%) is the predominant strain in Karachi. Venous and capillary blood are equally useful for malaria diagnosis however, convenience and less invasiveness may justify the preference of capillary blood over venous blood for early diagnosis of malaria. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3704 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3704/1307

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 351 361 EN Manuel Calvopina One Health Research Group, Faculty of Medicine, Universidad de las Américas (UDLA), Quito, Ecuador David Fonseca-Carrera Biotechnology Engineering, Faculty of Engineering and Applied Sciences (FICA), Universidad de las Américas (UDLA), Quito, Ecuador Irina Villacrés-Granda Biotechnology Engineering, Faculty of Engineering and Applied Sciences (FICA), Universidad de las Américas (UDLA), Quito, Ecuador Alberto Toapanta Biotechnology Engineering, Faculty of Engineering and Applied Sciences (FICA), Universidad de las Américas (UDLA), Quito, Ecuador Carlos Chiluisa-Guacho National Institute for Investigation in Public Health “Leopoldo Izquieta Pérez (INSPI)”, Tena, Ecuador Carlos Bastidas-Caldes Doctoral Program in Public and Animal Health, Universidad de Extremadura, Extremadura, Spain 2021 11 28 2023 09 22 Background: Leishmania is the parasitic protozoan responsible for leishmaniases, a disease that can cause a range of cutaneous, mucosal, and visceral infections. Two subgenera L. Viannia and L. Leishmania are known to infect humans in the tropics and subtropics of the Americas. The aim of the present study was to develop a new pair of primers for the two subgenera and test in clinical samples. Methods: We designed two new pairs of primers for a PCR method from two conserved genes, cysteine proteinase B (cpb) and N-acetylglucosamine-6-phosfate deacetylase-like protein (nagA), as specific markers for those two respective subgenera. Primers were tested with 16 microscopical positive clinical samples from the Amazon region of Ecuador obtained in 2010-2020 period. Results: The cpb presented a band of 172 bp and the nagA a band of 300 bp, thus clearly differentiating L. viannia from L. leishmania. Additionally, primers identified and differentiated the clinical samples in the two subgenera. Conclusion: The new primers targeting different two genes and standardized in a PCR assay could identified and differentiated Leishmania parasites at subgenus level. This protocol could be used for Leishmania genus identification and diagnosis at the subgenus level and for determining the parasite's geographical distribution where different Leishmania subgenera are found in the same area. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3513 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3513/1308

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 362 368 Farveh Ehya Department of Biotechnology, Tarbiat Modares University, Tehran, Iran Sima Kalantari 1. Digestive Oncology Research Center, Digestive Diseases Research Institute, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran 2. Regenerative Medicine Group (REMED), Universal Scientific Education & Research Network (USERN), Tehran, Iran Mojgan Bandehpour 1. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2. Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Bahram Kazemi 1. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2. Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2023 03 14 2023 09 22 Background: A lizard Leishmania has been isolated from a lizard (Agama agilis) in Iran. Its genome sequence has not been determined, so far. Methods: The study was done at Shahid Beheshti University of Medical Sciences, Tehran, Iran in 2017-2023. Leishmania promastigotes were cultured in RPMI1640 culture medium and collected at logarithmic phase by centrigugation. Parasite RNA was extracted by the Qiagene standard kit and its quantity and quality was determined and sequenced by NGS method with Illumina PE machine at BGI Company (China). Results: The number of 8316 mRNA, 83 tRNA, 63 rRNA, 83 ncRNA, 5 snRNA, 1039 snoRNA, 36 region, and 3 repeat regions, 8343 CDS, 9597 Exon and 9292 Genes were identified in promastigote of Iranian lizard Leishmania. Conclusion: Genomic elements of Iranian lizards Leishmania (with unique characteristics) were determined and identified by NGS system. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3902 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3902/1309

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 369 381 Serap Kılıç Altun Department of Food Hygiene and Technology, Faculty of Veterinary Medicine, Harran University, Şanlıurfa, Türkiye Mehmet Emin Aydemir Department of Food Hygiene and Technology, Faculty of Veterinary Medicine, Harran University, Şanlıurfa, Türkiye Sevil Alkan Department of Infectious Diseases and Clinical Microbiology, Çanakkale Onsekiz Mart University, Faculty of Medicine, Çanakkale, Türkiye Bünyamin İrehan Department of Parasitology, Faculty of Veterinary Medicine, University of Firat, Elazig, Türkiye 2023 06 20 2023 08 21 Background: Paragonimus is a genus of parasitic flatworms known as lung flukes that cause the parasitic disease paragonimiasis in humans and other mammals. We aimed to use bibliometric analysis to identify the global characteristics and temporal trends of published literature about paragonimiasis. Methods: Using the Web of Science database, we identified all original articles on paragonimiasis 1997 to 2022. After collecting the bibliographic and citation data, keywords, citation networks, and co-citations pertaining to paragonimiasis was carried out using the VOSviewer program. Results: The study identified 563 paragonimiasis articles published in 250 journals. Publications in paragonimiasis research have been cited 6190 times and 2803 times without self-citations. The years with the most publications were 2013, 2016, and 2021. The minimal threshold for analysis was met by 19 of the 52 countries investigated. The study included 19 items, yielding 170 links between countries. The total strength of these links was discovered to be 104772. The journal with the most publications in this category was Parasitology Research (n=31). The most frequently used terms in paragonimiasis study were "paragonimiasis", "Paragonimus westermanii", and "lung-fluke." Conclusion: The study concluded by providing an overview of the paragonimiasis research field, including current trends, development, and researcher collaboration. By addressing gaps in this bibliometric analysis and increasing collaboration, stakeholders could strengthen their strategies to effectively combat paragonimiasis and improve public health outcomes. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3987 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3987/1310

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 382 389 Guo Yue School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Province, Huzhou, Zhejiang, China Yu Liang Cao Emergency Department, South Taihu Hospital, Gangnan Road 1566, Huzhou, Zhejiang, China Dong Ying School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Prov-ince, Huzhou, Zhejiang, China Zhou Hongchang School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Prov-ince, Huzhou, Zhejiang, China Shang Ya Qian School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Prov-ince, Huzhou, Zhejiang, Chinaa Zhang De Long School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Prov-ince, Huzhou, Zhejiang, China Chen Gao Qiang School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Prov-ince, Huzhou, Zhejiang, China Jiang Bei School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Prov-ince, Huzhou, Zhejiang, China Wu Wan Ting School of Medicine, Huzhou University & Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Prov-ince, Huzhou, Zhejiang, China Lu Mingke School of Life Sciences, Xiamen University, Fujian, China 2023 02 08 2023 09 22 Background: The pathogen of angiostrongyliasis is the parasite Angiostrongylus cantonensis, and the transcriptome profiling of the male adult was unclear. We aimed to understand how the male adults adapt, so the expression profile of A. cantonensis adult males was analyzed. Methods: In order to improve the understanding of the transcriptome of adult males, RNA from three groups of male adult A. cantonensis was extracted and reverse transcribed to construct cDNA libraries. After sequencing, annotation of unigenes and transcripts was performed by querying the NR (Non-Redundant Protein Sequence Database), GO (Gene Ontology) and COG/ KOG (Clusters of Orthologous Groups of proteins/euKaryotic Ortholog Groups) databases. Results: For each group of adults, 43,260,894 raw reads and 43,200,341 clean reads were obtained. After successful assembly, 87,649 unigenes and 146,895 transcripts were obtained. Annotation of the unigenes and transcripts was identical and male adults expressed a series of genes encoding proteins specific to the male gender at the adult stage, such as proteins involved in energy metabolism, energy synthesis and transport. Expression of the ribosome pathway suggests a relationship with the physical activities during the adult male stage. Conclusion: The transcriptome analysis is a good reference to understand further the expression profile of male adult A. cantonensis. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3872 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3872/1311

Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 18 3 2023 09 22 390 399 EN Faezeh Hamidi 1. Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2. Department of Laboratory Sciences and Microbiology, Faculty of Medical Sciences, Tabriz Medical Sciences, Islamic Azad University, Tabriz, Iran Samira Mohammadi -Yeganeh 1. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Mostafa Haji Molla Hoseini Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Seyyed Javad Seyyed Tabaei Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Niloofar Taghipour Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Ameneh Koochaki Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Vahedeh Hosseini Department of Molecular Medicine and Genetics, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran Ali Haghighi Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2022 08 02 2023 09 22 Background: Immune cells and their secreted cytokines are known as the first barrier against pathogens. Leishmania major as an intracellular protozoan produces anti-inflammatory cytokines that lead to proliferation and survival of the parasite in the macrophages. miRNAs are small non-coding RNA molecules that regulate mRNAs expression. We aimed to investigate the relationship between the expression of TGF-β and a bioinformatically candidate miRNA, in leishmaniasis as a model of TGF-β overexpression. Methods: The miRNAs that target TGF-β -3´UTR were predicted and scored by bioinformatic tools. After cloning of TGF-β-3'UTR in psi-CHECK TM- 2 vector, targeting validation was confirmed using Luciferase assay. After miRNA mimic transfection, the expression of miR-27a, TGF-β, as well as Nitric Oxide concentration was evaluated. Results: miR-27a received the highest score for targeting TGF-β in bioinformatic predictions. Luciferase assay confirmed that miR-27a is targeting TGF-β-3'UTR, since miR-27a transfection decreased the luciferase activity. After miRNA transfection, TGF-β expression and Nitric Oxide concentration were declined in L. major infected macrophages. Conclusion: Bioinformatic prediction, luciferase assay, and miRNA transfection results showed that miR-27a targets TGF-β. Since miRNA and cytokine-base therapies are developing in infectious diseases, finding and validating miRNAs targeting regulatory cytokines can be a novel strategy for controlling and treating leishmaniasis. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/3696 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/3696/1312