Iranian Journal of Parasitology

PUF Proteins as Critical RNA-Binding Proteins in TriTryp Parasites: A Review Article

Tahereh Taheri — 2024-08-28

In eukaryotes, translation is a fundamental step in the long pathway of protein synthesis within the cell. In this process, several proteins and factors have involved directly or indirectly, individually or in association with other elements to contact mRNA. For perfect translation, many essential modifications should be done, such as cis-splicing to remove introns and two main events for capping and poly A polymerization in 5’ and 3’ end of mRNA, respectively. Gene expression is then regulated at both translation and stability of the target mRNA molecule levels. Pumilio/FBFs (PUFs) are the main group of RNA-binding proteins which bind to the 3’-UTR of target RNA and thereby regulate the fate, stability and subcellular localization of mRNAs and adjust the translated protein level. PUF proteins have been found both in nucleus where that bind to precursor mRNA, for processing and maturation of rRNA, and in cytoplasm where that bind to mRNA, stall the ribosomes, suppress the translation and localization of the mRNA. They can regulate the expression of mRNAs through activation or suppression of translation. Therefore, these proteins have recently garnered much attention as new generation of therapeutic targets against diseases such as cancer and neurological disorders. In comparison to other eukaryotes, trypanosomatids have a high number of PUF proteins, which function not only as gene expression regulatory factors but also in several biological processes such as differentiation and life-cycle progression of the cells. Here, we review the molecular and biological roles of known PUF proteins in TriTryp parasites (Trypanosome brucei, T. cruzi and Leishmania) beside some other parasites.

A New Immunogenic Structure of Polyepitopic Fusion against Leishmania major: In Silico Study

Saeed Pirmoradi — 2024-08-28

Background: The lack of complete protection against leishmaniasis and the challenges of anti-leishmaniasis drug treatment have made the treatment process more difficult. This study aimed to develop a new strategy for preparing a vaccine against cutaneous leishmaniasis using some of the antigenic proteins of the Leishmania parasite.

Methods: This study was carried out in 2022 at Shahid Chamran University of Ahvaz, Ahvaz, Iran. After preparing suitable epitopes of the Leishmania parasite and examining their antiparasitic properties, the process of making a fusion vaccine was performed and with the help of various bioinformatics tools, physicochemical and structural properties as well as immunological and simulation properties were studied and finally optimized. Construction and cloning were performed in the E.coli K12 system and finally, the docking process was performed with Toll-like receptors (TLRs), major histocompatibility complex I (MHC-I), and MHC-II receptors. With the help of selected epitopes of the Leishmania parasite, which had a high percentage of population coverage, a stable, antigenic, and non-allergenic chimeric vaccine was predicted.

Results: The results of the structural analysis of the TLR5\vaccine complex and simulation of its molecular dynamics showed a sufficiently stable binding. It also showed good potential for stimulation and production of active B cells and memory, as well as the potential for CD8+ T, CD4+ T cell production and development of Th2 and Th1-induced immune responses.

Conclusion: Computational results showed that the designed immunogenic structure has the potential to adequately stimulate cellular and humoral immune responses against Leishmania parasitic disease. As a result of evaluating the effectiveness of the candidate vaccine through in vivo and in vitro immunological tests, it can be suggested as a vaccine against Leishmania major.

Genetic Characteristics of Echinococcus granulosus from Fixed Paraffin-Embedded Tissue Samples in Human Isolates Based on the High-Resolution Melting Point Analysis in Sabzevar, Northeast Iran

Hossein Elyasi — 2024-08-28

Background: There are ten genotypes of Echinococcus granulosus with different intermediate and final hosts affecting the parasite's life cycle and its transmission to humans. Therefore, this study was conducted to determine the genotype of isolated hydatid cysts using the simple and fast high-resolution melting point analysis (HRM) method.

Methods: The paraffin tissue samples of patients who underwent surgery were obtained from the pathology sample bank of Vasei and Emdad Hospitals in Sabzevar, Iran during 2010-2020. The DNA content of the samples was extracted after collecting and determining the characteristics using the DNA extraction kit. PCR was performed on the samples and the presence of the hydatid cyst genome was confirmed using the special Master Kit. Mix PCR of Solis Biodyne Company and Real-Time device (Bio-Rad) were used, and the genetic identity of hydatid cysts were determined.

Results: Out of 33 paraffin samples, 21 samples contained hydatid cyst DNA, two of which were from the brain and 19 from the liver tissues; 12 samples did not contain hydatid cyst DNAs. All liver samples were from sheep species (G1), and the brain samples were from buffalo species (G3). Therefore, 9.53% of the Echinococcus species collected were buffalo (G3), and 90.47% were sheep (G1) strain.

Conclusion: Based on previous patterns, HRM methods can be used for easy and quick identification of Echinococcus strains. The G1 strain was the dominant strain causing hydatid cyst in different human organs, including the liver and brain.

Effectiveness of Miltefosine Nanoemulsion Concerning the Treatment of Acute and Chronic Toxoplasmosis: In Vivo Study

Gholam Reza Valizadeh — 2024-08-31

Background: Toxoplasma gondii is one of the most common parasites worldwide. It is of great importance to identify new potential drugs that are effective and less harmful in pregnant women and newborns. We investigated nanoemulsion miltefosine (NEM) in treating experimental acute and chronic toxoplasmosis.

Methods: A combination of triacetin, Tween 80, and ethanol (1:2) was used for the development of NEM formulations. The size of NEM was calculated to be 17.463 nm by DLS and TEM. To investigate the performance of miltefosine (MLF), NEM, sulfadiazine (SDZ), and pyrimethamine (PYR) (positive control) in vivo, acute toxoplasmosis was induced in mice by an intraperitoneal injection of RH strain tachyzoites. After five days, the mice were examined for the number and condition of tachyzoites and histopathological changes in the liver and spleen. Chronic toxoplasmosis was investigated in rats and the number and size of brain cysts along with histopathological changes were assessed in different groups.

Results: The results of the in vivo assessment of drugs in acute toxoplasmosis showed the following order regarding a decrease in the number of tachyzoites and an increase in survival rate: SDZ&PYR > NEM > MLF. The effects of drugs on chronic toxoplasmosis showed a significant effect of NEM (50%) on reducing the number of cysts compared to SDZ&PYR (10%) and MLF (12%) and reducing the size of NEM brain cysts (21%) compared to SDZ&PYR (5 %) and MLF (8%).

Conclusion: Increasing the penetration of NEM through the blood-brain barrier (BBB) and subsequently reducing the number and size of T. gondii tissue cysts is a promising new drug in treating chronic toxoplasmosis.

The Prevalence of Infection with Gastrointestinal Helminths in Free-Range Poultry of Zabol, Iran

Faezeh Sadeghi — 2024-09-01

Background: As an ecological and ethical method employed for poultry farming, free-range chicken keeping is growing and is important in fostering rural farmers' livelihoods and community prosperity. This study aimed to determine the nature and prevalence of helminth infections in native poultry in Zabol, Iran.

Methods: Between 2022 and 2023, 160 chickens were acquired and slaughtered ethically, and then their gastrointestinal tracts were taken and formalin preserved in Zabol, Iran. The parasites were isolated from gastrointestinal mucus in the laboratory and characterized using a microscope and specific methods such as Acetocarmine staining.

Results: Among 160 dissected birds, 92 (57.5%) were presented with gastrointestinal parasites, with 64 (40%) containing cestodes, 16 (10%) containing nematodes, and 12 (7.5%) infected by both. Observations were made of seven different species. Ascaridia galli (A. galli) (10%), Subulura brumpti (7.5%), and Heterakis gallinarum (3.12%) represent the three main nematode species. While Raillietina tetragona (R. tetragona) (33.12%) was the most prevalent cestode, R. echinobothrida (6.25%), R. magninumida (5%), and Cotugnia digonopora (3.12%) were less common.

Conclusion: The study revealed that free-range chickens in the arid Zabol region had a high prevalence of gastrointestinal parasites. There is a strong correlation between chicken foraging behavior and exposure to contaminated soil, with certain nematodes and cestodes constituting the most prevalent, negatively affecting bird growth, productivity, and health. Therefore, improving poultry welfare by applying parasite control measures and implementing appropriate management strategies is essential.

Feeding Behavior and Plasmodium Detection in Anopheles stephensi, a Ma-laria Vector in District Khyber, Khyber Pakhtunkhwa, Pakistan

Mahnoor Arif — 2024-09-01

Background: Anopheles stephensi is a significant malaria vector in Pakistan, and understanding its feeding behavior is necessary to control the spread of malaria. However, limited information is available on the host preferences of A. stephensi in Pakistan. Therefore, we aimed to explore the feeding behavior of A. stephensi, a malaria vector, in the District Khyber, Khyber Pakhtunkhwa, Pakistan.

Methods: A total of 7462 mosquitoes were collected between March and September 2021, with 1674 (22.4%) identified as A. stephensi (952 female and 722 male). Among the female A. stephensi, 495 (52%) were blood-fed. DNA was extracted from the blood-fed female A. stephensi mosquitoes using the Ammonium Acetate Precipitation Method followed by PCR analysis, blood meal sources were identified. Nested PCR on 191 pooled samples was used to detect Plasmodium falciparum and Plasmodium vivax.

Results: Cattle blood meals were predominant (73%), followed by human (20%) and chicken (7%), with no dog blood meals detected. All individual mosquito samples were negative for Plasmodium falciparum, while two pooled samples (out of 191) tested positive for P. vivax.

Conclusion: A. stephensi in Khyber District primarily displayed anthropophagic feeding behavior, with a small portion of the population infected with P. vivax. The results underscore the importance of targeted vector control strategies, environmental management, community engagement and continuous monitoring to suppress malaria transmission.

Seroprevalence of Strongyloides stercoralis Infection in Patients with Hyperlipidemia: A Case-Control Study

Mohammad Amin Sari — 2024-09-01

Background: Strongyloides stercoralis is one of the soil-transmitted helminths (STH) in tropical and subtropical regions. The role of lipid profile has been investigated in the survival of larval stages of helminths, but there is limited information about the role of lipid profiles and strongyloidiasis. Hence, we aimed to investigate the seroprevalence of S. stercoralis infection in patients with hyperlipidemia is compared with the non-hyperlipidemia.

Methods: In 2023, participants were selected from the laboratory of Porsina Hospital in Guilan Province, northern Iran and their lipid profiles including TG, CHOL, LDL, HDL, and VLDL were measured. They were divided into two groups of case and control and matched based on sex and age. S. stercoralis Ab (IgG) was measured by ELISA methods, using the NovaTec kit. Finally, statistical analysis was performed.

Results: Each case and control group consisted of 105 participants, from 13 to 80 years old. 56.66% were female and 43.33% were male. The sero-prevalence of S. stercoralis was found 4.76% in the case group compared to 0.95% in the control group. We found an association between TG fall and VLDL with sero-prevalence of S. stercoralis in hyperlipidemia group (P= 0.034), but other lipid profiles did not show a significant association. A significant relationship was found between contact with dogs and sero-prevalence of S. stercoralis (P=0.001).

Conclusion: The sero-prevalence of S. stercoralis in the case group was 5 times higher than the control group. A significant association between TG and VLDL fall with S. stercoralis Ab (IgG) was observed, but future studies with more sample sizes are suggested to investigate the anti-atherogenic effect of S. stercoralis. Also, a genetic assessment of S. stercoralis and the host (humane and dogs) is recommended to research zoonotic potential in epidemic areas.

Case Notification of Cutaneous Leishmaniasis at Different Elevations in the North-Central Ethiopia from 2018 To 2022

Bizuayehu Gashaw — 2024-09-01

Background: We aimed to analyze a four-year trend of Cutaneous leishmaniasis (CL) to determine risk levels and hotspots in North-central Ethiopia.

Methods: This retrospective study was conducted at Boru Meda Hospital (BMH) from March to April 2023, focusing on CL patients treated at the leishmaniasis treatment center (LTC). Data collected included age, gender, CL type, and other clinical factors. Each patient's origin was traced and geographically mapped by elevation to assess CL risk levels.

Results: There were a total of 573 CL patients reported from 46 districts, with a higher number of male patients (n=356) compared to female patients (n=217) (P <0.001). The median age of the patients was 21 years [15-30], with the highest number of CL cases observed among individuals aged 16 to 30 years. The majority of cases (69%) presented with localized CL (LCL). About 39% of patients had a previous treatment history for CL. A significant clustering of CL cases was observed at elevation of 2301-3300 meters above sea level (χ2:17.5; P <0.001), with the highest incidence (case notification) of 14.2/100,000 population.

Conclusion: Foci of CL, were burdened at higher elevations and no clinical variation were observed between elevation differences. The majority of cases were concentrated in an area covering approximately 21.4% of the total land mass. CL continues to be a significant issue in North-central Ethiopia and has the potential to spread to new areas.

Dirofilariasis in Male Genitalia: A Case Report and Literature Review

Rūta Žulpaitė — 2024-09-01

Human dirofilariasis is a rare anthropo-zoonotic disease, mainly detected in Southern and Eastern Europe, Asia Minor, Central Asia, and Sri Lanka. An increasing number of autochthonous Dirofilaria spp. infections has been recently reported in the areas previously considered free of the disease, including northern Europe and the Baltic States. A rare autochthonous case of scrotal dirofilariasis detected in Lithuania was described. Here, a 42-year-old male presented with a 1 cm nodule, limited in the scrotum. A nodule excision was performed. On histological examination, a degenerating roundworm with the features of Dirofilaria spp. (multilayered cuticle, well-developed musculature, focally preserved longitudinal ridges) was detected in the abscess cavity. No additional treatment was needed. Twenty-four publications reporting 28 male genitalia dirofilariasis cases in European countries have been identified.

Human Infection with the Nematode Gongylonema sp.: A Case Report from Bulgaria

Dimitar Vuchev — 2024-09-01

We report a case of infection with the zoonotic nematode Gongylonema sp. in a 24-year-old woman from Varna, Bulgaria. Two days before seeking medical attention, the patient had noticed a filamentous mass in her buccal mucosa that migrated when touched with the tongue. She manually removed a thread-like worm from the mucosa near her lower lip. Albendazole was prescribed for three days. The patient had no history of travelling abroad or contact with animals. To our knowledge, this is the sixth case that has been reported in the country.

Occurrence of Trypanosoma evansi Infection in Rottweiler Dog from Cauvery Delta Region of Tamil Nadu, India: A Case Report

Velusamy Rangasamy — 2024-09-01

Trypanosomes are the extraerythrocytic haemoflagellate protozoan parasites of domestic and wild animals including dogs. A male Rottweiler dog was presented with the history of anorexia, cloudy eyes and vomiting for a week. The dog was examined both physically and clinically. Blood was collected and subjected to haematological and biochemical analysis. Echocardiography of the thoracic cavity and ultrasonography of the abdominal cavity were performed. Molecular confirmation of parasite was carried out by PCR and DNA was sequenced to identify strain variations. Upon clinical examination, the dog revealed emaciation with high fever, unilateral corneal opacity, anemia, enlarged lymph nodes and distended abdomen. Echocardiography of the thoracic cavity showed ventricular hypertrophy and cardiomyopathy. Ultrasonography of the abdominal cavity showed splenomegaly and renomegaly. Haematological findings revealed decreased haemoglobin and erythrocyte count and biochemical findings revealed increased BUN and creatinine, hypoglycemia and hypoalbuminemia. Wet film and blood smear examination revealed actively motile trypanosomes and massive infection of trypanosomes, respectively. Molecular confirmation of species was carried out by amplification of 227 bp partial VSG gene of Trypanososma evansi, subsequently sequenced and phylogenitically analysed. The infected dog was treated intramuscularly with diminazene aceturate @ 3.5 mg /kg b.wt along with the supportive therapy. The study described the occurrence and diagnosis of trypanosomosis in dog. Early diagnosis and timely treatment is the most reliable way to safeguard the companion animals. It is the first report on trypanosomosis in dog from Cauvery delta region of Tamil Nadu, India.

Occurrence and Environmental Factors Associated with Cryptosporidium in South-Eastern Iran

Reza Shahraki — 2024-09-01

The Article Abstract is not available.

12th National and 5th International Congress of Parasitology and Parasitic Diseases of Iran (NICOPA XII), Karaj, Iran 2024

Mohammad Zibaei — 2024-09-01

The Article Abstract is not available.