Background: Vermamoeba vermiformis is one of the prevalent Free-living amoebae (FLA) in the environment. In recent years, studies regarding the distribution of V. vermiformis have increased, focusing on the importance of this FLA as a human pathogen but its pathogenic potential is still unclear. Moreover, the endosymbiosis relationship between V. vermiformis and microorganisms may lead to the development of multi-antibiotic drug resistance pathogens and may increase FLA virulence.
Methods: We aimed to perform a systematic review of the data on the occurrence of V. vermiformis in the environment and keratitis samples in Iran. Articles on the occurrence of V. vermiformis in environment and keratitis samples in Iran were systematically searched in PubMed, Scopus, and Google Scholar between January 2000 and July 2023. The keywords were carefully selected and accurately used to search for all relevant research articles to the title of the article. 
Results: Fourteen studies (n = 14) with a combined total of 436 environmental and clinical samples were included. The overall prevalence of V. vermiformis across all studies ranged from 2% to 50%, with the highest detection rates observed in drinking and surface waters.
Conclusion: Our findings suggest a wide distribution of V. vermiformis in water and different environmental sources in Iran. More comprehensive studies on the pathogenesis of this genus and the relationship between V. vermiformis and its endosymbiosis are research priorities.

Echinococcus granulosus sensu lato is a parasite that causes cystic echinococcosis in humans and herbivorous mammals, especially in the liver and lungs. The main treatment for cystic echinococcosis is surgery, but the primary risk of this method is the rupture of the cyst formed by the parasite and the spread of infectious protoscolices, leading to the formation of secondary cysts. Although many drugs are now used as scolicidal agents, their side effects have led to the acceptance of natural products as scolicidal agents. In this review, we aimed to investigate the scolicidal activities of various natural products. Studies on medicinal plants, herbal compounds, and other natural products with scolicidal activity against Echinococcus granulosus sensu lato protoscolices between 2000 and 2024 were searched using PubMed, Scopus, and Google Scholar databases. Information from the studies included the type of medicinal plant used, herbal compound or natural product information, concentration, exposure time, and scolicidal activity. This review analyzed 103 studies that assessed the efficacy of 60 natural products against Echinococcus granulosus sensu lato, including 39 medicinal plants, 15 herbal compounds, and six other natural products. The most frequently observed medicinal plants were from the Lamiaceae family, of which Zataria multiflora was the most studied species. In addition, thymol and propolis were the most commonly studied herbal compounds and other natural products.  This review focuses on medicinal plants, herbal compounds and other natural products with potential scolicidal agents.

Background: As the role of Blastocystis in the pathogenesis of intestinal disorders remains controversial, we aimed to clarify its clinical significance and subtype (ST) distribution in relation to colonoscopic findings.

Methods: Fecal samples were collected from adult patients scheduled for colonoscopy in Aydin Adnan Menderes University, Gastroenterology Department in 2023, 125 consecutive patients with colonic abnormalities and 125 with normal mucosa. Blastocystis positivity was determined with direct microscopy and amplification of the small subunit ribosomal RNA (SSU rRNA) gene. Subtypes were determined based on exact database matches. Demographic characteristics and clinical findings of participants were statistically analyzed.

Results: Blastocystis was detected in 26 fecal samples (10.4%) by direct microscopy and 32 samples (12.8%) by molecular test. Positivity was 16.0% (n=20) in controls and 9.6% (n=12) in patients with colonic abnormalities, with no significant difference (P>0.05). In males, Blastocystis was more prevalent in those with normal findings than with abnormalities (n=11, 19.6% vs. n=6, 7.7%, P<0.05), whereas no difference was observed in females (P>0.05). Subtype distribution was ST3 (59.4%, n=19), ST2 (21.8%, n=7), and ST1 (18.7%, n=6). No significant association was found between the overall Blastocystis positivity and age, gender, or gastrointestinal symptoms.

Conclusion: Although limited to routine colonoscopic findings, these results raise questions about the potential association of Blastocystis with a healthy gut or colonic abnormalities. ST3 was the predominant subtype, consistent with national and global reports.

Background: In immunocompromised individuals, Toxoplasma gondii infection can disseminate rapidly, causing severe cerebral and visceral complications. This study aimed to synthesize gold nanoparticles (AuNPs) via green synthesis and evaluate their therapeutic efficacy and mechanisms in immunosuppressed murine models of toxoplasmosis.

Methods: Mice were immunosuppressed using dexamethasone prior to infection with the Tehran strain of T. gondii. AuNPs were administered orally at 0.5 and 1 mg/kg, alone or combined with pyrimethamine (PYR; 10 mg/kg), in a 0.2 mL volume for 14 days. Survival rates were monitored daily. Brain cyst number and size were assessed microscopically. Oxidative stress markers, cytokine levels, and bradyzoite surface antigen 1 (BAG1) gene expression were analyzed by real-time PCR.

Results: Synthesized AuNPs exhibited cubic morphology with an average size of 20–30 nm. Combined AuNPs–PYR treatment significantly improved survival and reduced brain cyst burden (P=0.001). Treatment markedly decreased malondialdehyde (MDA) levels while enhancing glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity (P<0.01). Gene expression analysis demonstrated downregulation of BAG1 and IL-4 (<1.3-fold) alongside upregulation of IFN-γ and IL-12 (>4-fold). Hepatic and renal biomarkers were also significantly improved.

Conclusion: AuNPs, particularly in combination with PYR, demonstrate significant efficacy against toxoplasmosis in immunosuppressed murine models. Clinical trials are warranted to further assess the safety and therapeutic potential of this combination approach.

Background: Natural compounds have garnered increasing attention as candidate antiparasitic therapeutics. We aimed to evaluate the protoscolicidal efficacy of caryophyllene (CAP), alone and in combination with albendazole (ALZ), against hydatid cyst protoscoleces.
Methods: Suspensions of 10,000 invaginated and evaginated protoscoleces (400 µL normal saline) were exposed to CAP (62.5–500 µg/mL), ALZ (250 µg/mL), or CAP+ALZ combinations in 24-well culture plates. Protoscolex viability was assessed by eosin exclusion test. Reactive oxygen species (ROS) induction was quantified using the DCFH-DA assay, and pre-apoptotic gene expression was analyzed by real-time PCR.
Results: CAP, particularly in combination with ALZ, significantly reduced the viability of both invaginated and evaginated protoscoleces in a concentration- and time-dependent manner. Combination therapy achieved 100% mortality of invaginated protoscoleces within 10–30 minutes, demonstrating significant synergism over either monotherapy. Evaginated protoscolex viability was also markedly diminished. The observed scolicidal activity was mechanistically correlated with apoptosis induction, evidenced by elevated ROS production and significant upregulation of caspase-3 and P53 gene expression (P<0.001).
Conclusion: CAP, particularly in combination with ALZ, exhibits potent in vitro protoscolicidal activity against hydatid cyst protoscoleces, with apoptosis induction and ROS generation implicated as underlying mechanisms. Further pharmacodynamic investigations and clinical trials are warranted to elucidate the precise mechanisms of action and evaluate the therapeutic potential of CAP-based combination therapy in the management of cystic echinococcosis.

Background: Malaria remains a major global health burden, with increasing challenges posed by drug-resistant Plasmodium species. Another emerging threat is Plasmodium knowlesi, a zoonotic parasite responsible for rising human infections in Southeast Asia, particularly Malaysia. This study investigates the in vitro antimalarial activity of Silybum marianum (milk thistle) and Trigonella foenum-graecum (fenugreek) seed extracts against P. knowlesi, and their cytotoxicity against mammalian cells.
Methods: Using schizont maturation inhibition assay, P. knowlesi cultures were exposed to serial dilutions of both plant hydroethanolic extracts (0.20–100.00 μg/mL). Chloroquine was used as positive control. Cytotoxicity was evaluated on Vero cells using the MTT assay.
Results: Both extracts exhibited moderate antimalarial activity, with IC₅₀ values of 16.12 ± 1.02 μg/mL (S. marianum) and 17.08 ± 1.26 μg/mL (T. foenum-graecum). Cytotoxicity testing showed that the extracts were non-toxic to mammalian cells, with CC₅₀ values of 84.76 ± 15.12 μg/mL (S. marianum) and 72.75 ± 15.81 μg/mL (T. foenum-graecum). The selectivity indices were 5.26 and 4.26, respectively, indicating moderate selectivity towards the parasite over mammalian cells.
Conclusion: Both S. marianum and T. foenum-graecum demonstrate promising antimalarial potential against P. knowlesi in vitro, with acceptable cytotoxicity profiles. These findings support further investigation into their bioactive constituents and mechanism of action, offering preliminary evidence for their potential development as alternative antimalarial agents targeting zoonotic malaria.

Background: The Eurasian magpie, Pica pica (Linnaeus, 1758), is a widely distributed bird species inhabiting both rural and urban environments in Türkiye. Despite its ecological importance, information on its helminth fauna remains limited. We aimed to investigate the helminth fauna of P. pica specimens collected in Bursa Province and to contribute to the knowledge of parasite biodiversity in Türkiye.
Methods: A total of 53 P. pica specimens, brought dead between June 2015 and August 2025 from Bursa Zoo and the Celal Acar Wildlife Rescue and Rehabilitation Center (Karacabey Ova Korusu, Bursa, Türkiye), were examined for helminths using standard parasitological techniques. Recovered helminths were identified morphologically.
Results: Ten helminth species were identified: Brachylaima fuscata (Rudolphi, 1819) Joyeux, Baer & Timon-David, 1932, B. lobatum (Railliet, 1900), Lyperosomum longicauda (Rudolphi, 1809) (Digenea); Raillietina echinobothrida (Megnin, 1881), Passerilepis stylosa (Rudolphi, 1809), Choanotaenia macracantha (Krabbe, 1869) (Cestoda); Dispharynx nasuta (Rudolphi, 1819), Baruscapillaria obsignata (Madsen, 1945) Moravec, 1982, B. resecta (Dujardin, 1845) (Nematoda); and Sphaerirostris picae (Rudolphi, 1819) (Acanthocephala). R. echinobothrida, C. macracantha, B. obsignata, and B. resecta were recorded from P. pica for the first time worldwide. In addition, B. fuscata, B. lobatum, L. longicauda, C. macracantha, P. stylosa, and B. resecta represent new records for the helminth fauna of Türkiye.
Conclusion: This study expands current knowledge of the helminth fauna of P. pica and provides several novel host and geographical records. The findings highlight the importance of continued parasitological investigations of wild birds in Türkiye to improve our understanding of parasite diversity and host–parasite associations.

Background: Cutaneous leishmaniasis is a neglected parasitic disease prevalent in tropical and subtropical regions, caused by different species of the genus Leishmania. Current therapeutic options are limited due to toxicity, prolonged treatment regimens, and emerging drug resistance.

Methods: This study was conducted at Kashan University of Medical Sciences, Kashan, Iran in 2024. The antileishmanial activity of quercetin and a synthesized quercetin–ZnMgFe₂O₄ nanostructure was evaluated against L. major promastigotes in vitro conditions. The nanostructure was synthesized and characterized using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The antileishmanial activity was assessed by determining the mortality rate of promastigotes at different concentrations (150, 300, 450, and 600 μg/mL) of quercetin and also quercetin–ZnMgFe₂O₄ nanostructure. In addition, ultrastructural and nuclear alterations were examined following exposure to 150 and 600 μg/mL of both quercetin and the nanostructure.

Results: The quercetin–ZnMgFe₂O₄ nanostructure exhibited significantly higher inhibitory activity compared with quercetin. The mortality rates for the nanostructure were 73% and 85.9% at 150 and 600 μg/mL, respectively, compared with 42% and 81% for quercetin. Microscopic analysis revealed pronounced morphological alterations, including damage to the cell body and flagella, as well as destruction of the nucleus and kinetoplast.

Conclusion: The Quercetin_ZnMgFe₂O₄ nanostructure exhibited considerable antileishmanial activity against L.major promastigotes in vitro. Considering the obtained results, this nanostructure can be considered in anti-leishmanial studies and further in vitro and in vivo investigations are required to evaluate its safety and efficacy.

Background: The efficient management of schistosomiasis depends on the rapid and accurate diagnosis of infection. Rapid diagnostic tests are essential for identifying morbidity markers of urogenital schistosomiasis.
Methods: A total of 1,341 urine samples collected from school-age pupils in Kwara State were screened for Schistosoma haematobium eggs using the syringe filtration method. Haematuria and proteinuria were assessed using commercial urine reagent strips (Medi-Test Combur-9).
Results: Overall, 277 paticipants were infected, of which children >15 yr accounted for 37.5% of the disease prevalence. Only light and moderate infection was observed. Microhaematuria varied significantly with egg intensity (P<0.05), but no association was found between hematuria severity and age. The overall prevalence of macrohaematuria was 1.3%, while microhaematuria and proteinuria were 53.9% and 19%, respectively. Macrohaematuria was found in 88.9% of S. haematobium-positive children while 13% of positive children tested negative for microhaematuria. Microhaematuria showed higher sensitivity (70.8%) compared to other biomarkers, while macrohaematuria had higher specificity (99.8%).
Conclusion: In the absence of other morbidity markers, microhaematuria appears to be a good proxy for urogenital schistosomiasis in our setting.

Background: We investigated the prevalence of Toxoplasma gondii in slaughtered livestock (sheep, goats, camels) in Sabzevar, Eastern Iran using nested PCR methods.

Methods: Samples of diaphragm and heart tissue were collected from 40 sheep, 40 goats, and 40 camels at local slaughterhouses. DNA was extracted from the samples and analyzed using nested PCR targeting the B1 gene of T. gondii.

Results: The results indicate an overall prevalence of 60% (24/40) in sheep, with 37.5% (15/40) of diaphragm samples and 22.5% (9/40) of heart samples testing positive. In goats, the overall prevalence was 52.5% (21/40), with 35% (14/40) of diaphragm and 17.5% (7/40) of heart samples positive. Camels showed the highest prevalence at 65% (26/40), with 45% (18/40) of diaphragm and 20% (8/40) of heart samples infected. Diaphragm tissue showed higher infection rates compared to the heart in all species. No significant difference was found in infection rates between male and female animals. RFLP analysis using XhoI on selected positive PCR products differentiated genotypes based on digestion patterns. Among analyzed samples (19 per species), genotypes II/III were identified in 57.9% (11/19) of sheep, 36.8% (7/19) of goats, and 47.4% (9/19) of camels, with the remainder likely genotype I or undigested II/III.

Conclusion: The high prevalence rates observed, especially in diaphragm tissue and in camels, have important implications for public health and food safety. Enhanced meat inspection, public education, and further research on effective control measures are recommended.

Background: We aimed to determine the prevalence of Entamoeba gingivalis and Trichomonas tenax in relation to periodontitis in Urmia, Northwestern Iran, in 2022.

 Methods: Gingival crevicular fluid (GCF) samples were collected from133 patients and analyzed using microscopic and molecular techniques to identify parasites. Data were analyzed using SPSS software and Chi-square and Fisher's exact test.

Results: 42.1% of patients were infected with E. gingivalis and 11.3% with T. tenax. In comparison, microscopic diagnosis identified only 1.5% and 0.8% of patients infected, respectively. This highlights the increased sensitivity of molecular methods for parasite detection. A statistically significant relationship was observed between the presence of these parasites (identified by the molecular method) and several factors. Male patients had a significantly higher prevalence of E. gingivalis infection (P=0.037). Additionally, a history of diabetes (P=0.011) and cardiovascular diseases (P=0.021) was associated with a higher prevalence of E. gingivalis. The highest prevalence of E. gingivalis was observed in the 41–50-year age group (15 cases). Importantly, both E. gingivalis and T. tenax were significantly more prevalent in patients with severe periodontitis (P=0.003, P<0.001).

Conclusion: This study demonstrates a higher prevalence of E. gingivalis compared to T. tenax in the oral cavity, suggesting its potentially greater role in periodontal disease, particularly among individuals with specific risk factors. Further research is warranted to elucidate the precise contribution of these parasites to periodontitis, identify additional risk factors, and develop improved diagnostic and treatment approaches.

Background: Blastocystis spp. is a globally distributed intestinal protozoan that infects humans and various animals. This protozoan is a common pathogen among humans and animals and can be transmitted from animal species to humans. Therefore, the present study aimed to identify and characterize Blastocystis subtypes in children with gastrointestinal symptoms using PCR and sequencing.

Methods: Overall, 150 stool samples were collected from children referred to academic hospitals in Mashhad, Iran over a period of seven months in 2020. Participants were divided into two groups: symptomatic cases (n = 50) and asymptomatic controls (n = 100). DNA was extracted from Blastocystis-positive samples and analyzed by PCR followed by sequencing for subtype identification.

Results: Blastocystis spp. was found in 6 (12%) of the symptomatic children and 7 (7%) of the asymptomatic group. All isolates from symptomatic cases belonged to subtype ST1, whereas isolates from the control group included ST1, ST2, ST3, and ST7. ST1 was the most frequently detected subtype overall. Among symptomatic children, abdominal pain was the most frequent symptom.

Conclusion: ST1 was the predominant subtype in both symptomatic and asymptomatic children, though it was the only subtype observed in symptomatic cases. These findings suggest a potential link between ST1 and gastrointestinal symptoms. Further studies with larger populations and in diverse geographic regions are warranted to investigate the clinical significance and distribution patterns of Blastocystis subtypes.

Background: In Algeria, leishmaniasis is characterized by an endemic nature, which appears in humans in the two known forms: visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL). This study aimed to determine the seroprevalence of canine leishmaniasis within a group of dogs and the incidence of human leishmaniasis.

Methods: We performed a serological survey of canine leishmaniasis (CanL) in 84 asymptomatic dogs in Constantine wilaya (north-west of Algeria). The collected dog sera were tested by the indirect ELISA method. The study considered multiple risk factors, including the breed, sex, age, and location of the dogs. The dogs whose blood samples were taken during this survey came from the following municipalities in Constantine: El-Khroub, Ali Mendjeli, Zighoud Youcef, Aïn Smara, Hamma Bouziane, Didouche Mourad, and Constantine. In parallel, an epidemiological study of human leishmaniasis in the Constantine region was conducted. We collected retrospective data on the two forms of human leishmaniasis from Constantine's Department of Public Health between 2016 and 2024.

Results: The incidence of CanL observed in the study was 8.33% (with 7 dogs testing positive. During the nine-year study period, 136 cases of CL and VL were recorded. The predominance of CL (with 124 cases, 91.18%) over VL (12 cases, 8.82%) was statistically significant (χ²=92.24; P<0.001) throughout the study duration.

Conclusion: The high seroprevalence of leishmaniasis in asymptomatic owned dogs, along with the significant frequency of human cases of CL and VL in the Constantine region, indicated that leishmaniasis remained a major public health problem in Algeria.

Background: The recent increase in unresponsive cases to current treatments poses a significant challenge in the effort to discover new antileishmanial drugs. We aimed to investigate the effects of Mn₂O₃ NPs alone or in combination with shark cartilage extract (ShCE) on L. infantum (MCAN/ES198/LIM-877) both in vitro and in vivo.

Methods: The susceptibility of promastigotes and amastigotes was evaluated using MTT and microscopy assays with different concentrations of Mn₂O₃ NPs alone or in combination with ShCE (Mn₂O₃-ShCE). The combination index (CI) was used to assess potential synergy. Cytotoxicity against J774.A1 macrophages was measured, and the selectivity index (SI) was calculated. Also, apoptosis was analyzed in promastigotes. The therapeutic and prophylactic effects were tested in BALB/c mice, and survival rates were monitored.

Results: Mn₂O₃ NPs caused a dose-dependent decrease in proliferation of promastigotes (IC₅₀=19.8±1.1 μg/mL) and amastigotes (IC₅₀=29.9±0.05 μg/mL). Co‑treatment with ShCE significantly enhanced the antileishmanial activity, reducing the IC₅₀ values of promastigotes and amastigotes. The CC₅₀ values were 159.9±4.2 μg/mL for Mn₂O₃ NPs and 276.5±7.9 μg/mL for Mn₂O₃-ShCE. Apoptosis rates in promastigotes treated with Mn₂O₃ and Mn₂O₃-ShCE were 80.7% and 86.3%, respectively. Parasite loads in the tissues significantly decreased (P<0.001) in BALB/c mice treated with Mn₂O₃ NPs, and survival rates increased significantly (P<0.05). The combination with ShCE further improved outcomes, with 100% survival observed in the Mn₂O₃-ShCE group (P<0.01).

Conclusion: These findings demonstrated a synergistic effect between Mn₂O₃ NPs and ShCE against L. infantum. Therefore, it may serve as a promising candidate for future development of leishmanicidal drugs.

Background: We investigated Dientamoeba fragilis molecular distribution and genetic types among Nineveh Governorate patients with symptoms through SSU rRNA gene PCR amplification and sequencing.

Methods: Fifty stool samples were collected from symptomatic patients aged 3–68 years attending the Medical Research Hospital in Nineveh Governorate, northern Iraq in 2024. Genomic DNA was extracted using the Presto™ Stool DNA Extraction Kit. The PCR reaction used species-specific primers DF400/DF1250 to produce ~850 bp amplicons. Positive products were sequenced and examined using BLAST, MEGA 11, DnaSP, and PopART. The Maximum Likelihood method used the Tamura 3-parameter model to perform phylogenetic analysis. PCR detection indicated D. fragilis in 12/50 stool samples, corresponding to a prevalence of 24%.

Results: The BLAST analysis showed that the sequence had 97%-99.7% similarity to global reference strains (AY730405). The study identified three haplotypes which contained three mutations while showing a haplotype diversity (Hd) of 0.318 that indicated minimal genetic diversity. The phylogenetic analysis showed that all isolated strains belonged to genotype 1 but U37461 formed a separate lineage as genotype 2. The Iraqi isolates showed sequence similarity to genotype 1 reference strains reported globally.

Conclusion: This study provides additional molecular evidence of D. fragilis in northern Iraq and the second report nationally. Genotype 1 was identified among all analyzed isolates with minimal genetic variation between different groups. The molecular detection methods delivered vital diagnostic data but scientists need to expand their surveillance activities to study animal disease transmission to humans and animal disease progression.

Background: We aimed to present the morphometric analysis of adult Brachylaima sp. infect wild pigeons, Columba livia, in the United Arab Emirates (UAE).

Methods: Brachylaima sp. specimens were collected from the intestinal tracts of 5 C. livia in Al Ain City, UAE, from 2023 to 2024. After extraction and preservation in 70% ethanol, 15 specimens were stained with acetocarmine, dehydrated, cleared, and permanently mounted for detailed microscopic analysis. Their morphology was systematically measured and compared to existing data for other Brachylaima species.

Results: The adult Brachylaima sp. revealed an elongated cylindrical body with rounded ends. Oral and ventral suckers were equally sized and muscular, with the oral sucker at the anterior end and the ventral sucker in the anterior body quarter. A short pre-pharynx led to a spherical pharynx, with no observed esophagus. Tubular digestive ceca bifurcated posterior to the pharynx, extending laterally to the body’s terminus. Two oval, tandem testes occupied the posterior quarter, larger than the ovary positioned between them. A tubular cirrus pouch and seminal vesicle lay anterior to the anterior testis. A small seminal receptacle adjoined the ovary’s posterior margin. The uterus formed a convoluted loop ascending toward the pharynx before descending to a genital pore near the anterior testis, filled with ovoid operculated eggs. Vitelline follicles spanned bilaterally from the ventral sucker to the anterior testis. The excretory ducts are connected to a tubular vesicle between the ceca, ending in a subterminal pore.

Conclusion: Given the medical importance of Brachylaima, further experimental and molecular studies are critical to resolve its taxonomy, life cycles, and transmission dynamics in arid ecosystems.

No Abstract.

Onchocerciasis is a chronic helminth disease in which morbidity accumulates slowly over years through repeated cycles of microfilarial production, immune activation, and tissue injury. Adult worms of Onchocerca volvulus can survive for more than a decade within the human host, while pathology arises primarily from inflammatory responses to dying microfilariae rather than parasite mass itself. Although large-scale ivermectin programmes have reduced transmission and blindness, individual disease trajectories remain highly variable, and cumulative skin, ocular, and neurological damage may persist despite treatment. A digital twin framework for onchocerciasis, defined as a continuously updated virtual representation integrating parasite longevity, treatment cycles, immune responses, and tissue damage, offers a novel approach to modelling lifelong disease burden. By combining longitudinal clinical data, treatment history, immunological markers, and exposure context, such a system could predict disability trajectories, optimise treatment timing, and support precision strategies for morbidity prevention. Here, we describe the conceptual basis, biological architecture, and translational potential of an onchocerciasis digital twin.