Background: Apoptosis, a determined form of programmed cell death (PCD), occurs in multi-cellular and single-celled organisms. Given that a general understanding of apoptosis in single-cell Leishmania is crucial for designing disease control policies, we reviewed the apoptosis mechanism and the proteins involved.

Methods: The information was obtained from articles published in PubMed, SciELO, Science Direct, Scopus, Google Scholar, and Web of Science databases (1998-2021). Search terms used were^” apoptosis^” or^” Leishmaniasis^”.

Results: The 77 subjects were included in the study that revealed the significance of the apoptosis process for Leishmania survival. Although, various stimuli induce Leishmania apoptosis, the proteins involved in apoptosis have been poorly understood. Metacaspases in Leishmania instead of caspase and death receptors in mammals play the same role in the PCD pathways. Also, other apoptotic proteins in Leishmania such as endonuclease G (EndoG), caspases- like cysteine proteases, TSN (Tudor Staphylococcal Nuclease), and Zinnia endonuclease 1 (ZEN1) lead to phenotype similar to mammalian apoptosis. Furthermore, there are differences in these mechanisms between the different species of Leishmania and studies to illustrate downstream events related to the serine phosphatidylcholine exposure, cytochrome C secretion, etc. remain an ongoing challenge.

Conclusion: Determining the essential regulatory proteins in the Leishmania apoptosis and the specific present of metacaspases in parasite, is effective for designing new therapeutic strategies against leishmaniasis and vaccine development.

Background: Cutaneous Leishmaniasis (CL) is a highly infectious parasitic disease in Iran. Although various diagnostic methods are available, identifying a sensitive and specific approach remains essential for effective treatment and disease management. We aimed to compare microscopy, the Leishmanin Skin Test (LST), and the Direct Agglutination Test (DAT) for diagnosis of human CL.
Methods: Fifty samples were obtained from clinically suspected CL cases. The results of conventional methods, including microscopy, the LST, and the DAT, were then compared among these samples. For microscopical examination considered a gold standard, smears were stained with Giemsa 10% and then examined for the observation of amastigote forms for the LST, 0.1 ml of standard Leishmanin solution was intradermally injected into positive patients, and then indurations were measured after 48-72 hours; an induration of ≥5 mm was considered positive. Additionally, anti-Leishmania antibodies were detected using DAT on positive collected serum samples.
Results: Out of 50 samples collected from individuals suspected of CL, 66% (33/50) of them showed positive results using microscopic examination. Among these 33 patients diagnosed with CL, 9% (3/33) tested positive in the LST, and only 1 (3%) patient showed specific antibodies against   Leishmania using DAT.
Conclusion: The microscopy method appears to be more suitable for diagnosing CL. However, there is a clear need for additional diagnostic methods with more validity for CL.

Abstract

Background: Dicrocoeliasis is an important helminthic zoonosis reported from many parts of the world. Due to low-performance medications, drug delivery is a great challenge in improving the treatment of this liver fluke infection. We aimed to determine the anthelmintic properties of Nanosilver oxide (AgO) against Dicrocoelium dendriticum infection.
Methods: The impacts of various concentrations of AgO nanoparticles (50-200 µg/ml) for 12-24 hours were compared with closantel, a chemical drug. The anthelmintic efficacy was evaluated using the scanning electron microscopy (SEM) technique. The synthesized nanoparticles were analyzed for structural assessment using XRD, UV–VIS spectroscopy, and SEM. The XRD pattern shows the formation of AgO nanoparticles.
Results: The UV-VIS spectra showed the broad peak, corresponding to Ag nanoparticles. SEM images of treated parasites by AgO (200 µg/ml) showed severe damage, which includes complete loss of sensory papillae and destruction of prominent network structures and tegument vesicles. The mortality rate increases with the increase in the concentration and exposure time of the parasite to nanoparticles. Besides the MTT assay, the toxicity of AgO, at concentrations of 800 µg/ml was 8.7%.
Conclusion: AgO NPs have potent anthelmintic effects on liver fluke D. dendriticum. This is the first research that assessed the effect of AgO NP on liver fluke D. dendriticum. Hence, the present study provides a basis for future research on the control of this common trematode.

Background: We aimed to investigate the serological and molecular characteristics of Toxoplasma gondii infection among pregnant women and their aborted placentas in Meshkin-Shahr City during 2019-2020.
Methods: Blood samples of 210 pregnant women were evaluated for anti-T. gondii antibodies and related risk factors were determined. Also, the sera of aborted women and their buffy coats and aborted placenta tissues were used to detect anti-Toxoplasma antibodies and the parasite’s DNA, respectively. The parasite genotypes were determined by the PCR-RFLP using the SAG3 gene.
 Results: The overall prevalence of anti-Toxoplasma IgG was 24.3% and only 1% of participants were seropositive for the IgM antibody. There was a significant relationship between raw or unwashed vegetable consumption, contact with soil, vegetable/fruit washing type, and seropositivity (P<0.05). During pregnancy, 4.7% of women encountered an abortion and 30% and 50% of cases were positive for IgG antibodies before and after abortion, respectively. Only two cases were IgM seropositive after abortion. In the avidity IgG test, 20% of cases showed low avidity. BLAST and phylogenetic analysis exhibited that all isolates belonged to the type III T. gondii genotype. Although two women with spontaneous abortions showed seropositivity for IgM T. gondii antibody, parasite DNA was detected in three cases.
Conclusion: The seroprevalence of Toxoplasma infection is not high in pregnant women. Seropositive women are not safe from congenital transmission. T. gondii type III is the etiology of fetus infection in mothers with spontaneous abortion. It seems that screening and essential care are still necessary during pregnancy.

Background: We aimed to investigate the effect of chitosan nanoparticles loaded azithromycin on reducing the number of Toxoplasma gondii tissue cysts in the brain of a mouse model.
Methods: Chitosan nanoparticles and azithromycin loaded nanoparticles were synthesized using microfluidic system and characterized using dynamic light scattering (DLS) and TEM images. Forty BALB/c mice after infection with a cyst forming T. gondii strain, were divided into four groups daily receiving PBS, 10 mg/kg azithromycin, 10 mg/kg chitosan nanoparticles, and 10 mg/kg chitosan nanoparticles loaded azithromycin, respectively, for 10 days. Immediately after end of the treatment, the mice were sacrificed and the tissue cyst burden in their brain was investigated using an optical microscope and compared by ANOVA statistical test.
Results: The average particle size and dispersion index for chitosan nanoparticles were 193.66 nm and 0.43, and for nanoparticles containing azithromycin drug, they were 233.66 nm and 0.21, respectively. The amount of drug loading was 1.8% and the drug release was more than 90% after less than 48 hours. The stability of nanoparticles did not change significantly after 28 days of observation. Toxoplasma tissue cyst numbers obtained in a range of 1.48 to 1.95 in 10 ul brain suspension with no significant differences among the groups of treated mice.
Conclusion: The synthesis of chitosan nanoparticles loaded with azithromycin by microfluidic system could make the particles with more uniformity and stability and high loading of the drug with low cost and more convenient conditions.

Background: Leishmania aethiopica is the leading cause of cutaneous leishmaniasis (CL) in Ethiopia. Different clinical manifestations might be related to host immunity, which itself can be influenced by the host's nutritional status. However, there is limited evidence that associates nutritional status with CL in Ethiopia. We investigated the relationship between clinical variables of CL and malnutrition.
Methods: A retrospective study was conducted in June 2024. Patient data was analyzed from those treated for CL and screened for nutrition from January 2022 to May 2024 at Tefera Hailu and Addis Zemen Primary Hospitals. Nutritional status was assessed through Anthropometric measurements.
Results:  A total of 470 CL patients were treated, with a prevalence of 14.65/100, 000 population affected.  Out of the total CL patients, 217 were assessed for nutrition, 22% were malnourished. Malnutrition was most prevalent in mucosal (30%) and recurrent cases (38.5%), compared to localized (20%) and new cases (21%) respectively.
Conclusions:  Malnutrition might have the potential to shape the clinical manifestation and treatment outcome in CL patients. In CL endemic areas nutritional supplement with the treatment of CL could require for better patient outcome.

Background: Iron deficiency anemia (IDA) is a common health issue during pregnancy and may be influenced by parasitic infections such as Blastocystis hominis. This study aims to assess the role of Blastocystis hominis infection as a potential risk factor for IDA in pregnant women.
Methods: A total of 208 pregnant women were enrolled in this cross-sectional study, including 98 women with IDA (case group) and 110 women without IDA (control group), conducted at Imam Reza and Ghaem Hospitals in Mashhad, Iran during 2022–2023. Hemoglobin and ferritin levels were measured in all participants, with anemia defined as hemoglobin <11 g/dL and ferritin <15 μg/L. Stool samples were collected from all participants to detect B. hominis and other parasitic infections using direct smear, formalin-ether concentration, and modified acid-fast staining methods. Statistical analyses were performed using independent samples t-test and chi-square test, with a significance level of P<0.05.
Results: The prevalence of B. hominis infection was significantly higher in the case group (38.8%) compared to the control group (10.9%) (P<0.001). Hemoglobin and ferritin levels were significantly lower in the case group (P<0.001). However, no significant difference was observed between the groups in terms of socioeconomic status, education, or occupation.
Conclusion: B. hominis infection may contribute to iron deficiency anemia in pregnant women. Screening for parasitic infections in anemic pregnant women, particularly B. hominis, could improve anemia management during pregnancy.

Background: Approximately 25% of hepatic hydatid cysts rupture into the biliary tract. The precise effect of bile within the cyst on protoscoleces remains unclear. We aimed to elucidate the effect of bile on protoscoleces.
Methods: The contents of hydatid cysts from the livers of three sheep were aspirated under sterile conditions. The aspirated contents were divided into 50 separate Eppendorf tubes (5 cc). Samples from each tube were stained with 0.1% eosin Y. Pink stained protoscoleces were considered dead under light microscopy (x100). A total of 100 protoscoleces were counted in each sample, and the number of live and dead protoscoleces was recorded. The tubes were randomly divided into five groups. Group 1 served as the control, Group 2 received normal saline (NS), Group 3, received hypertonic saline, Group 4 received bile, and Group 4 received diluted bile. The number of live and dead protoscoleces was recorded at the end of the first and second hours.
Results: Compared to the initial count of live protoscoleces, the number of live protoscoleces increased at hours 1 and 2 in Groups 2 and 4. No live protoscoleces remained at hours 1 and 2 in Group 3. There was no significant change in Group 5. When compared to the control group, a significant increase in viability was observed only in Group 4 (P=0.001).
Conclusion: Bile of sheep does not exhibit scolicidal effects; rather, it positively affects protoscoleces by increasing viability.

Background: Medicinal plants and their derivatives due to having high availability, low cost, low toxicity, and high efficacy are recognized as significant reservoirs of a diverse array of valuable therapeutic compounds. We aimed to investigate the lethal effects of carvone (CV) on protoscoleces of hydatid cyst, in laboratory and ex vivo conditions.
Methods: The protoscolicidal effects of CV were investigated both in vitro and ex vivo on hydatid cyst protoscoleces using the eosin exclusion assay. Furthermore, the impact of CV on the induction of apoptosis and DNA damage in the protoscoleces was assessed through a colorimetric protease assay and Real-time PCR analysis, respectively.
Results: CV, particularly at a concentration of 150 μg/ml, effectively eradicated protoscoleces of hydatid cysts within a 20-minute exposure period. Furthermore, CV exhibited sustained anti-parasitic effects in the ex vivo, leading to the complete elimination of hydatid cyst protoscoleces after a 30-minute exposure, in contrast to its immediate lethal effects observed in vitro. CV concentrations of 1/3 IC₅₀, 1/2 IC₅₀, and IC₅₀ (P < 0.001) resulted in caspase-3 activation levels of 11.3%, 19.8%, and 28.4%, respectively. The expression levels of the EgATM and EgP53 genes significantly upregulated after treatment with CV (P<0.001).
Conclusion: The findings demonstrated the potential of CV to eliminate protoscoleces by inducing apoptosis and causing DNA damage. However, further studies are required to clarify the specific mechanisms underlying its action and to assess its efficacy in clinical trials, which may facilitate the application of CV in the context of hydatid cyst surgical procedures.

Background: Given the significant role of chitosan nanoparticles in medicine, the present study aimed to assess the in vivo efficacy of synthesized chitosan nanoparticles coated with thymol (CNCT) in combating Toxoplasma gondii infection.
Methods: Mice were administered CNCT orally at dosages ranging from 0.25 to 0.75 mg/kg/day for a duration of 14 days. Following this treatment, they were infected with T. gondii tachyzoites of the Rh strain to induce acute toxoplasmosis. Then, the mortality rate, parasite load, antioxidant activity, and the gene expression level of proinflammatory cytokines were evaluated.
Results: The dimensions of CNTN exhibit variability, with a mean size of 295 nm. The prophylactic administration of CNTN in mice infected with T. gondii resulted in a significant enhancement in survival rates and a considerable decrease in parasite load (P<0.001). The CNTN caused a significant decrease in malondialdehyde level, while a notable increase (P < 0.001) in the activity of the antioxidant enzymes. The feeding the mice infected with CNTN caused a meaningful elevation in the expression level of TNFα- and IL-1β (P<0.001). The biochemical analyses indicated no significant changes in the serum levels of liver and kidney function markers.
Conclusion: The recent study revealed that CNTN demonstrates promising in vivo effects against toxoplasmosis in murine models. These effects are attributed to its antioxidant properties and immunomodulatory capabilities, which increase specific pro-inflammatory cytokines without any noticeable signs of toxicity to liver and kidney function.

Background: Helminthic infections cause helminthiasis, including infections by Strongyloides stercoralis, a kind of helminths that cause reinfection and lead to severe infections, can be transmitted through the soil. We aimed to identify S. stercoralis and other helminthic infections in rubber tree plantations in Thailand's lower northern regions. The specific goals include assessing prevalence using Formalin Ethyl–acetate Concentration Technique (FECT) and Agar Plate Culture (APC) and constructing S. stercoralis phylogenetic tree.
Methods: Overall, 646 fecal samples from rubber plantation workers in five provinces in northern Thailand were examined using FECT and APC under microscope. DNA from larvae confirmed as Strongyloides spp. by Polymerase Chain Reaction (PCR) was sequenced for phylogenetic analysis. The DNA sequences were also submitted to the GenBank database.
Results: Prevalence of helminthic infections was 8.82%, with soil transmitted helminths (STH) prevalence at 6.81%; S. stercoralis accounted for 5.41%, with Ascaris lumbricoides at 0.62%, hookworm 0.46%, and T. trichiura 0.31%. PCR analysis successfully amplified the 18S rRNA gene in 26 out of 34 genomic DNAs, indicating a detection rate of 70.59%. Sequencing of these PCR products identified S. stercoralis strains closely related to those reported in the Republic of Lao, Myanmar, and Japan, suggesting genetic diversity within the species.
Conclusion: STH prevalence, predominantly S. stercoralis, highlights public health concerns in rubber plantation areas, necessitating enhanced monitoring and intervention strategies. Phylogenetic analysis of S. stercoralis, revealing a close genetic relationship among strains from various Southeast Asian countries, which underscores potential patterns of transmission and evolutionary relationships in the regions.

Background: We aimed to determine the seroprevalence of Trichinella in horses in West Azerbaijan Province, Iran a region known for its wildlife interactions.
Methods: The study was conducted in March 2016 across four cities in West Azerbaijan: Urmia, Chaldoran, Sardasht, and Khoy, Iran. A total of 184 equines, ranging from three to over ten years of age, were randomly sampled. Blood samples were collected and were analyzed using the ID Screen® Trichinella Indirect Multi-species ELISA to detect antibodies.
Results: Of the 184 serum samples, 4 (4.3%) from northern counties and 2 (2.2%) from southern counties tested positive for trichinellosis. Chi-square and Fisher's exact tests indicated no significant correlations between age, gender, or residential location and disease prevalence. The logistic regression model was not statistically significant (P = 0.754), revealing that these demographic factors do not substantially influence trichinellosis risk in the studied equines.
Conclusion: This study provides crucial insights into the low seroprevalence of Trichinella in horses in West Azerbaijan, suggesting that equines may not be primary reservoirs of the parasite despite overlapping habitats with infected wildlife.

Background: We aimed to investigate the prevalence of Enterocytozoon bieneusi and the circulating genotypes in wild boar in the western regions of Iran.
Methods: Fifty-two fecal samples were collected from wild boars in Lorestan province, Iran. After DNA extraction, the presence of E. bieneusi was evaluated by real-time PCR. A nested PCR targeting the internal transcribed region (ITS) was employed to characterize genotypes. The PCR products were sequenced, and the genetic diversity and relationships among the genotypes were identified using MEGA X and DnaSp (V5) software.
Results: Sixteen (30%) samples were positive for E. bieneusi using real-time PCR, and 11 (21%) were positive for nested PCR, which was sequenced. All 11 positive samples were identified as the BEB6 genotype (Group 2). The haplotype diversity was noted to be 0.182, and the nucleotide diversity, calculated using DnaSp, was 0. 00085.
Conclusion: The findings highlight the high prevalence of the genotype BEB6 in wild boars. The presence of this genotype suggests the circulation of E. bieneusi between domesticated animals and wild boars in Iran.

Background: Theoretically, there is a possible association between emerging SARS‑CoV‑2 infection and parasites such as Toxoplasma gondii. We aimed to evaluate the seroepidemiology of T. gondii in COVID-19 patients and the control group as well as its correlation with risk factors.
Methods: Totally, 450 sera samples were taken from COVID-19 positive patients and controls from the Tehran, Karaj, and Shiraz cities, Iran. Anti-T. gondii IgG and IgM were evaluated using the ELISA technique. After two months, the participants were followed for recovery or non-recovery and even death. The association between seroprevalence and severity of viral infection as well as other risk factors was statistically estimated.
Results: IgG prevalence in patients and healthy individuals was 59.11% and 61.77%, respectively; these values were estimated at 2.22% and 0% for IgM, respectively. There was no significant association between the prevalence of IgG with COVID-19 infection, while this association was statistically significant for IgM prevalence. The Karaj had the highest prevalence, and a significant association was observed between the seroprevalence and some variables.
Conclusion: Despite the non-significant association between the chronic phase of T. gondii (sero) prevalence and COVID-19 symptomatic forms, the parasite prevalence was estimated remarkable and the viral infection and parasite-related acute phase antibodies relationship was estimated to be statistically significant. Due to immunosuppressive therapies for this viral inflammatory infection, it makes it more difficult to interpret the results, and because of the vulnerability of the immune system of these individuals, toxoplasmosis is likely to be hazardous in them; therefore, screening for this parasitic complication seems necessary.

Cryptosporidium species are a significant cause of persistent untreatable diarrhea in HIV-positive patients, especially in developing countries; however, extra-intestinal instances are rarely documented. Herein, we present three cases of pulmonary cryptosporidiosis in HIV patients at Imam Khomeini Hospital, Tehran, Iran. Despite the combination treatment with antiretroviral therapy, azithromycin, and paromomycin, the death of all 3 patients occurred due to severe complications. These cases highlight the importance of early diagnosis and management of respiratory cryptosporidiosis in HIV patients with low CD4 counts.

Myiasis infection is caused by the invasion of flies' eggs and larvae into different tissues of the host's body. The rarest form is urogenital myiasis, which can occur even in men with a much lower probability than women, especially in tropical and subtropical regions. The case of this study is a 5-year-old boy from suburbs of Bushehr City, southern Iran who referred to the Pediatrics Department of Persian Gulf Hospital in Bushehr due to constant fever and significant weight loss. In urine evaluation, the presence of Chrysomya bezziana larvae was confirmed. The larvae of several species of myiasis-producing flies can attack the urinary-genital system of people and cause urogenital symptoms. Therefore, it is necessary to diagnose and accurately identify the species of larvae that cause myiasis in order to properly treat affected patients.

No Abstract. Not Abstract.