Background: We aimed to review and analyze the status of Medical Parasitology course including current problems and challenges on human resources, infrastructure, research capacities, educational and investigative priorities in Iran, and to develop a roadmap and future development in this field.

Methods: A questionnaire was prepared to obtain information on Medical Parasitology departments in the country followed by field visits in many cases to collect data on the status of the faculty members, laboratory experts, physical spaces, laboratory equipment, microscopic slides, training Parasitology techniques, research projects and theses, published articles and books, access to Parasitology journals, books and e-books during 2022-2023. A roadmap and future development for Medical Parasitology in Iran and the formulation of a desirable status concerning short term to long-term goals was compiled.

Results: While prominent strengths such as capable research laboratories, experienced faculty, suitable educational spaces, active research centers, the existence of centers for keeping and working with laboratory animals, international and national collaborations are seen in a number of medium to large centers, shortcomings related to expert human resources, lack of educational equipment and slides, the need to upgrade especially in practical subjects, journal clubs, and lab meetings, research infrastructure such as laboratory animal houses, budget for theses and research projects urgently need attention. Among 132 universities, faculties, and educational-research centers training personnel in allied medical sciences disciplines, a considerable number lack Parasitology specialists.

Conclusion: Iran has a strong foundation in Medical Parasitology. However, using non-permanent faculty paid per course in medical education undermines quality and strains resources, exacerbating shortages. Universities must implement tailored strategic plans for parasitology, regularly updated to strengthen education, research, and health services, ensuring sustained excellence in training and public health contributions.

Background: Leishmania major is an obligate and intracellular pathogen and the macrophages are the cell hosts for L. major. Imiquimod stimulates macrophages to secrete different cytokines via the expression of TLRs.
Methods: This study was carried out in the Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, in 2018. The effect of imiquimod was investigated on non-infected and infected macrophages with L. major on the expression of Toll-like receptors (TLRs) and inflammatory cytokines. TLRs play an important role in enhancing the proceeding of phagocytosis and killing parasites. Moreover, the cytokines such as TNFα, IL6, and IL1, are often identified in inflammatory conditions as interfering targets in treatment. Healthy macrophages and macrophages infected with Leishmania major parasites were affected by different concentrations of imiquimod, after that the expression of TLR genes (TLR1, TLR2, TLR3, TLR4, TLR7 and TLR9) and cytokines were evaluated by real time RT-PCR. For experiments in laboratory animals, infected BALB/c mice were exposed to imiquimod and then isolated peritoneal macrophages.
Results: The expression of TLR2 decreased in non-infected macrophages were affected by the imiquimod. The expression level of TLR7 in healthy macrophages, decreased and the difference with control group was significant. Imiquimod increases the expression of inflammatory cytokines and IL12 in mouse macrophages and also decrease the expression of IL10.
Conclusion: This suggests that imiquimod may improve the therapeutic effects in infected mice with Leishmania major. Imiquimod causes that TLR2 decreased expression but TLR7 and TLR9 increased expression. Imiquimod as TLR7 agonist, enhance the recovery of leishmaniasis.

Background: Cerebral cystic echinococcosis (CCE) and coenurosis are zoonotic diseases caused by the larval stages of Echinococcus granulosus sensu lato (s.l.) and Taenia spp., respectively. Due to the similarity between the symptoms and clinical samples of CCE and cerebral coenurosis, especially in cases with no protoscoleces, the diagnostic methods for the differentiation of CCE from cerebral coenurosis are crucial, especially in countries where both diseases are endemic. To compare CCE and coenurosis, morphometric indices of protoscoleces and molecular methods were used in the present study.
Methods: In this regard, four isolates of human cerebral echinococcal cysts, three isolates of Coenurus cerebralis from sheep, and one non-cerebral Coenurus from sheep muscles were evaluated. The isolated specimens have been collected from Shiraz, Ahvaz, Tehran and Kerman from before 2000 to 2022. The molecular characterization was carried out using the partial NADH dehydrogenase1 (nad1) gene. Phylogenetic analysis was performed using the maximum likelihood method.
Results: In fertile cysts, the total size of the large and small hooks of Coenurus was larger than cerebral echinococcal cyst. These parameters demonstrated significant morphological differences between the C. cerebralis and the cerebral echinococcal cyst. Molecular methods identified the cerebral echinococcal cysts as E. canadensis (G6) genotype. One C. cerebralis and the non-cerebral Coenurus were identified as Taenia multiceps and T. multiceps gaigeri, respectively.
Conclusion: Morphometric indices are significantly different between protoscoleces of C. cerebralis and cerebral echinococcal cysts. Hence, they could be used for differential diagnosis of the fertile cysts of these cestodes. However, in cases with no protoscoleces, molecular methods are essential for the differentiation of CCE from cerebral coenurosis, especially in regions where both diseases are prevalent and endemic.

Background: We aimed to conduct a comprehensive analysis of cutaneous leishmaniasis’s (CL) clinical polymorphism by examining the relationship between clinical forms, duration of illness, and their spatiotemporal distribution.
Methods: A retrospective study at University of Gondar Hospital analyzed cutaneous leishmaniasis patients treated from 2022 to 2024. Demographic and epidemiologic data were collected, with parasite detection via microscopic and clinical examination.
Results: Overall, 454 CL cases from 49 districts were diagnosed, predominantly affecting males aged ≤30, with a mean age of 25.31 yr (SD ±18.3). Significant differences were noted in age and sex (P<0.05). Approximately 70% had lesions ≥4 cm. Most CL cases had 2+ parasite loads. Sodium stibogluconate (SSG) remained the primary treatment choice for patients. The spatial distribution of CL cases covered a larger geographical area, although the cases (>20) were concentrated in Central Gondar. The mucosal CL shared a similar geographical pattern with the recurrent CL type. Notably, 48% had chronic presentations and lived with the disease for ≥12 months. In diffuse CL (DCL) a longer delay was seen and its clinical presentation was associated with longitudinal time series. Acute patients exhibited a higher parasitic load than chronic ones (38% vs. 24%), respectively.
Conclusion: CL significantly affected school-aged children. The symmetrical distribution of cases in districts studied could increase the attention of clinicians and enhance management strategies. Extended disease durations necessitated specialized treatments for clinical transitions.

Background: The use of nanocarriers in combination with other treatments shows significant promise in addressing drug-resistant diseases, particularly malaria. Given the high prevalence of drug-resistant malaria, research into innovative therapies is crucial. This study focuses on a nanoform of chitosan, a biodegradable polymer, combined with Crocus sativus (saffron) and chloroquine to enhance their antimalarial effects.
Methods: Saffron extract and chloroquine were separately conjugated with chitosan, followed by confirmation tests to determine conjugation efficiency. Both chloroquine-resistant and sensitive strains of Plasmodium falciparum were cultured to calculate the IC50 values of various treatments in vitro. This study was conducted at the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2024.
Results: Confirmation tests (FTIR, DLS, Zeta potential, TEM) verified proper drug conjugation to nanocomposites, with observed nanosize, the percentage of conjugation was 64.4% for chloroquine and 42.9% for saffron. Toxicity and hemolysis tests confirmed safe doses. The IC50s values for Chloroquine, Nanoparticle-Chloroquine, Saffron, and Nanoparticle-Saffron were 0.3, 0.8, 42.5, and 6.24 µg/ml, respectively, for the sensitive strain, and 5, 1, 12.5, and 3.12 µg/ml, respectively, for the resistant strain. Combination therapy with the fixed ratio method showed synergistic effects. Statistical analysis revealed synthesized nanocomposites' superior inhibition of P. falciparum growth compared to non-nano. Significant differences were observed in some cases (P< 0.05).
Conclusion: Utilizing nanocarriers and combination therapy is an appropriate strategy for addressing drug resistance. Saffron's anti-malarial effects on P. falciparum were notably increased when linked to chitosan nanocomposites. Furthermore, employing a fixed ratio technique enhanced the therapeutic effectiveness of saffron when combined with chloroquine and chloroquine-nanocomposites across all concentrations.

Background: Toxoplasmosis and toxocariasis are two zoonotic diseases with global impact. Chronic kidney disease (CKD) can lead to complications, associated with reduced immune responses that predispose them to frequent parasitic infections. We aimed to determine the prevalence of toxoplasmosis and toxocariasis in CKD patients to propose a new way to control them in dialyses duration in Tehran, Iran.
Methods: Three hundred and sixty-five CKD patients and 72 healthy individuals were tested for anti-Toxoplasma gondii (IgG, IgM) and anti-Toxocara (IgG) antibodies using conventional ELISA technique. IgM positive samples underwent genetic analysis.
Results: Of the 437 samples studied (365 patients and 72 controls), 182 CKD patients (49.8%) and 20 controls (27.9%) were positive for Toxoplasma IgG, and 8 (2.2%) CKD samples were positive for Toxoplasma IgM, while none of the control samples were positive for Toxoplasma IgM. Molecular analysis of the 8 IgM positive samples with B1gene confirmed the presence of Toxoplasma antigen in all of them. Toxocara IgG antibodies showed a lower prevalence in CKD patients (5.7%) compared to controls (9.7%). The study revealed significant differences (P <0.05) in Toxoplasma IgG and IgM antibodies, as well as variables such as dialysis and dialysis duration between the two groups of patients and the control group. However, variables such as Toxocara IgG, age, gender, lupus, and pets showed no significant difference between the control group and the patients.
Conclusion: The collected data in this study could serve as a reference for future studies and may be useful for examination and evaluation of acute and chronic toxoplasmosis before and after starting dialysis.

Background: The present study was carried out to investigate the prevalence of intestinal microsporidia in patients with intellectual disabilities in the rehabilitation centers of Hormozgan Province in south of Iran, using microscopic and molecular methods.
Methods: The current cross-sectional study was conducted on 119 stool samples obtained from patients with intellectual disabilities. Acid-fast trichrome staining was used to detect parasites. Moreover, all of the stool samples were assessed for the presence of microsporidia DNA by the PCR method.
Results:  The mean age of the subjects was 29 (±12.3) years. All stool samples were examined for microsporidia infection, and only one case of Enterocytozoon was detected in one of the samples. The results of the nested-PCR test on 119 stool samples showed one case of infection with Enterocytozoon bieneusi, corresponding to a prevalence of 0.84%. The nested-PCR test for Encephalitozoon intestinalis detected no infections in any of the stool samples.
Conclusion: The overall prevalence of microsporidia among patients with intellectual disabilities in Hormozgan rehabilitation centers is low and limited to E. bieneusi.

Background: We aimed to analyze the presence of Entamoeba histolytica lectin light chain antigen and p53 protein in colorectal cancer biopsies to uncover potential antigenic interactions between them.
Methods: Overall, 150 colorectal cancer biopsy samples were subjected to examination for the specific E. histolytica antigen and p53 protein through the application of the immunohistochemical technique. To augment diagnostic precision, the western blotting method was employed and its results were compared with those obtained from immunohistochemistry (IHC).
Results: Among the 150 colorectal cancer biopsy samples examined, 100 cases tested positive for the p53 protein, while 19 cases exhibited positivity for the E. histolytica light chain lectin antigen. Notably, only 14 cases demonstrated a positive presence for both the p53 protein and the E. histolytica protozoan.
Conclusion:  Entamoeba histolytica, a leading cause of parasitic infections, raises questions about its potential link to colorectal cancer, specifically in connection with the p53 protein. The findings presented in this investigation signal the need for further research to comprehensively understand the implications of E. histolytica in colorectal cancer diagnosis and treatment strategies.

Background: Among the many Entamoeba species that infect humans, only Entamoeba histolytica is considered pathogenic, being responsible for amoebiasis or amoebic dysentery.
Methods: Between June and October 2022, a total of 106 stool samples were collected from children under six years of age presenting with diarrhea at Paiji Hospital in the city of Paiji, Iraq. DNA was extracted from all stool specimens to detect the presence of parasitic organisms.
Results: Of the 106 fecal samples, 4 (3.7%) tested positive for Entamoeba spp. using an initial PCR amplification targeting approximately 900 bp of the 18S rRNA gene. Among these, only one sample tested positive for E. histolytica using a nested multiplex PCR assay. In this study, neither E. moshkovskii nor E. dispar was detected. Sequence analysis of the partial 18S rRNA gene revealed that 0.9% of samples were positive for E. histolytica, while 2.8% were positive for E. coli. The sequences were deposited in GenBank under the accession numbers OP868733.1 for E. histolytica and OP868730.1, OP868731.1, and OP868732.1 for E. coli.
Conclusion: Children were infected with different species of Entamoeba. Molecular methods are essential for distinguishing between Entamoeba species due to their significance in accurate diagnosis and appropriate treatment strategies.

Background: Cytauxzoon felis is a protozoan parasite transmitted by ticks and affects felids. Acute infection in domestic cats is characterized by symptoms such as lethargy, anorexia, fever, and anemia.
Methods: The present study focuses on diagnosing and molecularly identifying C. felis using a real-time PCR method in cats from Semnan, Iran. During the winter and spring of 2024, two hundred cats were randomly selected from veterinary clinics in Semnan. Blood samples were collected from the cats for DNA extraction and molecular analysis. Samples were divided into 40 pooled of 5 samples, each consisting of a combination of 5 blood samples. Then, the genomic DNAs were extracted from blood specimens and screened by real-time PCR (RT-PCR) for the presence of C. felis infection by amplifying of ITS2 gene belonging to the Cytauxzoon genus.
Results: The results indicated that 6 out of the 200 blood samples were infected (3%).
Conclusion: This study was conducted for the first time in Semnan and shows that the prevalence of C. felis in cats is significant.

Background: We investigated the contributing factors of amoebic dysentery across five provinces in Northeastern Thailand from 2019 to 2023. Comprehensive data were collected regarding agricultural practices, disaster impacts, and demographic characteristics for Sisaket, Ubon Ratchathani, Yasothorn, Amnat Charoen, and Mukdahan.
Methods: Overall, 1,112 cases of amoebic dysentery were analyzed, revealing a higher prevalence among males in younger age groups, while females dominated in older cohorts. Sisaket reported the highest number of cases, followed by Ubon Ratchathani, Amnat Charoen, Yasothorn, and Mukdahan. Notably, Sisaket demonstrated the highest incidence rate of 10.28 cases per 100,000 population, compared with Ubon Ratchathani's 5.27 cases.
Results: The study highlights the interplay between environmental factors, agricultural practices, and sociodemographic characteristics, emphasizing how high disaster impact and reliance on natural agriculture may facilitate amoebic dysentery transmission. The findings underscore the importance of tailored public health interventions to mitigate the disease burden, particularly in vulnerable populations. Future research should focus on pinpointing specific factors driving the observed variations in disease prevalence across the region.
Conclusion: This study serves as an essential resource for policymakers and public health officials aiming to enhance amoebic dysentery management strategies in Northeastern Thailand.

Background: The Mediterranean form of visceral leishmaniasis (VL) is endemic in some regions of Iran and is often seen in children under 10 years old. There is a 90% mortality in patients, if diagnosis and treatment are not done on time. Canids, as reservoirs, play an important role in the spread of the disease.
Methods: Bushehr Province, southern Iran is always mentioned as one of the endemic areas for VL, so for this purpose, as the first study in the region, 112 sheltered dogs in Bushehr City were evaluated for canine leishmaniasis (CanL) using serological Direct Agglutination Test (DAT) as well as detailed clinical examinations.
Results: Out of 112 samples collected, 71.4% of cases had anti-L. infantum IgG antibody titers of 1:80 and higher and therefore considered as seropositive. Furthermore, from the 70 seropositive dogs with antibody titer of 1:320 and higher, 47 (42%) had at least one of the clinical symptoms associated with VL and considered as cases with CanL.
Conclusion: The current seroprevalence situation of dogs in this region, is very noticeable and can be an important alarm for policymakers and health system practitioners. More comprehensive and complementary parasitological studies should be carried out on a number of reservoirs in the region for diagnosis and treatment and to accurately determine the statistics of the disease compared to the obtained seroprevalence status.

Background: Stunting, resulting from chronic malnutrition, increases susceptibility to infections due to immature immunity. Blastocystis hominis and Giardia duodenalis may contribute to stunting. We aimed to determine the characteristics of intestinal protozoan infection among stunting children.
Methods: A cross-sectional study was conducted in 2020 among 280 stunted children in Bandung Regency, West Java, Indonesia. Faecal specimens were collected, with portions preserved separately in 10% formaldehyde and RNA Later solution. Of these, 230 met the examination criteria. Risk factors and demographic data were obtained through interviews. DNA was extracted, and intestinal protozoan infection were detected using PCR targeting the 18S SSU rRNA gene for B. hominis and 16S-like RNA gene for G. duodenalis.
Results: The prevalence of G. duodenalis and B. hominis was 5.6% (13/230) and 55.6% (128/230), respectively. Multivariate analysis identified age (P-value 0.004; OR 0.327) and no availability of a septic tank (P-value 0.021; OR 4.881) were the significant risk factors for G. duodenalis infection. For B. hominis infection, significant risk factors included age (P-value 0.033; OR 0.722) and gender (P-value 0.047; OR 1.742).
Conclusion: Stunting and intestinal protozoan infection present a dual burden. G. duodenalis and B. hominis infections were prevalent among stunted children. Significant risk factors included age and septic tank unavailability for G. duodenalis, while age and gender were associated with B. hominis infection. Improved sanitation and targeted interventions are essential to reduce infection risks.

Background: Respiratory infections caused by Lophomonas blattarum and Pneumocystis jirovecii are significant threats, especially to immunocompromised patients. Both pathogens are associated with severe pneumonia and are often underdiagnosed due to the challenges in identifying them accurately, particularly in co-infections. We aimed to evaluate the prevalence and clinical impact of co-infections with L. blattarum and P. jirovecii in patients with respiratory symptoms.
Methods: This cross-sectional study involved 111 patients admitted to the Pulmonary Ward of Imam Reza Hospital in Mashhad, Iran in 2023. Bronchoalveolar lavage (BAL) samples were collected from all patients and analyzed microscopically and molecularly. PCR amplification targeting L. blattarum and P. jirovecii was performed, with subsequent sequencing for molecular identification. The presence of Pneumocystis was identified using a 346-bp PCR band.
Results: Of the 111 patients, Lophomonas was detected in 48 patients (43.2%), and Pneumocystis in 47 patients (42.3%). Co-infections were identified in 26 patients (23.6%). Both infections were more common in males, though the difference between genders was not statistically significant. The highest prevalence was observed in patients over 60 years, with 18% and 19.8% infection rates for Lophomonas and Pneumocystis, respectively. Co-infection rates were significantly higher in older patients and in males (P= 0.028).
Conclusion: The study demonstrates a significant prevalence of co-infections with L. blattarum and P. jirovecii in patients with respiratory conditions, particularly in the elderly. The findings underscore the need for comprehensive diagnostic strategies, including molecular and microscopic approaches, to accurately diagnose and manage these co-infections in high-risk populations.

Background: We aimed to investigate the presence of thyroid peroxidase antibodies (anti-TPO) in women with spontaneous abortion, both with and without toxoplasmosis. This was achieved by evaluating and comparing the serum levels of interleukins IL-4, IL-6, IL-10, IL-17, and TNF-α in women who experienced abortion due to toxoplasmosis, categorized by positive and negative anti-TPO status, with those who were toxoplasmosis-negative and also negative for anti-TPO.
Methods: We evaluated the serological presence of IgG and IgM antibodies to Toxoplasma using an ELISA method, in Samarra City, Salah al-Din Governorate, Iraq in 2021-2022. A sample of 153 women with spontaneous abortion were enrolled. We also measured the serum levels of interleukins IL-4, IL-6, IL-10, IL-17, TNF-α, and anti-TPO using the same technique.
Results: Overall, 103 were Toxoplasma-positive. Of these, 14 had positive anti-TPO results (13.5%), compared to only 3 positive cases among the 50 matched controls who were Toxoplasma-negative (6.0%). The difference between Toxoplasma-positive and Toxoplasma-negative women regarding anti-TPO status was statistically significant for interleukins IL-4 (P=0.010), IL-6 (P=0.017), and IL-10 (P=0.003), but not for IL-17 or TNF-α. Additionally, the statistical analysis revealed a highly significant difference in the average concentrations of interleukins IL-4 (P=0.013) and IL-10 (P<0.001) between the Toxoplasma-positive/anti-TPO-positive group and the Toxoplasma-negative/anti-TPO-negative group of aborted women.
Conclusion: Elevated concentrations of IL-4, IL-10, and IL-6 have been associated with women undergoing recurrent miscarriages and negative anti-TPO results. The complex interaction between pro-inflammatory and anti-inflammatory cytokines is essential for immunological balance and pregnancy outcomes in a condition of toxoplasmosis. An increase in IL-4 and IL-10 levels in anti-TPO-positive individuals may lead to an imbalance in immune response, facilitating the development of autoimmune thyroid disease.

Malaria continues to pose a significant public health challenge, particularly in certain regions of Indonesia, where it remains endemic. Plasmodium falciparum is responsible for the most severe form of the disease, often leading to life-threatening complications such as acute kidney injury (AKI). Here, we report the case of a 22-year-old male from Sikka Regency, East Nusa Tenggara, Indonesia, with a seven-day history of intermittent fever following recent travel to malaria-endemic areas. On physical examination, he appeared somnolent and exhibited icteric sclera, hepatomegaly, and dark yellow urine. Laboratory findings were notable for impaired kidney function (serum creatinine 3.52 mg/dL (311 μmol/L)), elevated transaminases, hyperbilirubinemia, thrombocytopenia, and a P. falciparum parasitemia level of 9.7%. Imaging studies revealed pulmonary edema, enlarged kidneys, ascites, pleural effusion, and hepatomegaly. The patient was diagnosed with severe falciparum malaria, complicated by AKI, pulmonary edema, and jaundice. He was then treated with intravenous artesunate for six days, followed by a three-day course of oral dihydroartemisinin/piperaquine and a single dose of primaquine. Additionally, he underwent two sessions of timely hemodialysis. His clinical condition and kidney function gradually improved thereafter, and he was discharged without sequelae. This case highlights that early diagnosis and appropriate treatment can lead to full recovery from AKI caused by severe P. falciparum malaria.

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