Sciomyzidae (Diptera) larvae live as predators and/or parasitoids of snails. They have potential economic importance as biological control agents against gastropods that are vectors of parasitic diseases. In the present study, the use of Sepedon larvae in schistosomiasis control was evaluated by studying the influence of environmental parameters on snail distribution and the impact of prey duration and availability on the survival of Sepedon (P) ruficeps and Sepedon (S) nasuta larvae. The predatory capacity of the larvae was also evaluated by determining the influence of snail consumption on the longevity of the different larval stages. The study was carried out from August 2014 to July 2015 in three locations. A total of 3,448 specimens were collected, showing high diversity and abundance, with a significant difference (df = 2, c2 = 582.97; P < 0.05) among locations. The snail distribution was influenced by environmental variables (total hardness, electric conductivity, Ca[2+], O2, Mg[2+]). The predator/prey interaction between Sciomyzidae species and snails was examined. No difference in the mean duration of immature stages and a mean predation capacity of the two flies (P = 0.50) vs. (P = 0.31) respectively for Sepedon (P) ruficeps and Sepedon (S) nasuta. The consumption varied considerably depending on the ratio of predator (larva) to prey (snail) size and the larval stage. Both these Sciomyzidae consume healthy or parasitized snails by schistosome larvae indiscriminately. They appear to be equivalent agents for biological control of freshwater snails which are intermediate hosts, involved in the transmission of human schistosomiasis or veterinary distomatosis.

Cystic echinococcosis (CE) is a globally distributed parasitic zoonosis caused by infection with Echinococcus granulosus. This study investigated the ferroptosis mechanism in hepatocyte injury induced by Echinococcus granulosus cyst fluid (EgCF) and its relationship with the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway using an in vitro model. We employed the Cell Counting Kit-8(CCK-8) assay, Calcein AM/PI staining, and flow cytometry to assess cell viability, reactive oxygen species (ROS) production, and mitochondrial membrane potential (MMP). Additionally, we measured lipid peroxides, malondialdehyde (MDA), glutathione (GSH), and ferrous iron (Fe[2+]) levels. Western blot analysis examined the expression of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11(SLC7A11), prostaglandin-endoperoxide synthase 2 (Ptgs2), ferritin heavy chain (FTH1), Nrf2, and heme oxygenase-1 (HO-1). Results indicated that EgCF inhibited hepatocyte proliferation and promoted hepatocyte apoptosis while inducing ferroptosis. Treatment with ferroptosis inhibitors Deferoxamine (DFO) or N-acetylcysteine (NAC) significantly attenuated EgCF-induced cytotoxicity. Nrf2 activation markedly mitigated EgCF-induced hepatocyte injury. This study elucidates the role of EgCF-induced ferroptosis and Nrf2 signaling in CE-associated liver injury, providing novel theoretical insights and new targets for disease prevention and treatment strategies.

In eukaryotes, with the exception of mammals, trehalose fulfills multiple biological functions, including energy provision and stress tolerance. Trehalose-6-phosphate phosphatase (TPP) is a key enzyme in the trehalose biosynthetic pathway, rather than a trehalose synthase per se. Loss of TPP function or knockdown of its expression impairs trehalose synthesis, which in turn leads to developmental abnormalities or lethality in nematodes. These observations suggest that TPP represents a potential target for the development of anti-parasitic therapeutics. This study focused on the cloning, expression, and characterization of Trichinella spiralis TPP (TsTPP). A recombinant plasmid was constructed to express and purify the recombinant TsTPP protein (rTsTPP), whose immunogenicity was confirmed using positive serum from Trichinella spiralis (T. spiralis)-infected mouse. The expression of TsTPP at different developmental stages of T. spiralis was determined by immunofluorescence assay. The specific TPP inhibitor N-(phenylthio) phthalimide (NPP) was employed to evaluate its lethal effect on T. spiralis muscle larvae (ML) in vitro. Furthermore, molecular methods such as qPCR and Western blot were used to detect the transcription and protein expression levels of TsTPP upon NPP treatment, aiming to identify new drug targets against T. spiralis. We successfully constructed the recombinant plasmid and obtained rTsTPP. Positive serum assays demonstrated the immunogenicity of rTsTPP, and we used indirect immunofluorescence assay (IFA) to determine its localization across different developmental stages of T. spiralis. Distinct distribution patterns were observed: TsTPP was diffusely distributed throughout the body of newborn larvae (NBL), accumulated in the intestinal glands, midgut ampulla and genital primordia of muscle larvae (ML), and localized to the mid and posterior intestine of adult worms (AW). To verify the lethal effect of NPP in vitro, we treated ML with NPP at concentrations of 12.5, 25, 50, 100, and 200 µg/mL for 24, 36, and 48 h, and evaluated its lethal effect by statistical analysis of worm morphology. The experimental results showed that the mortality rate of ML gradually increased with increasing NPP concentration and culture time. When the concentration was 200 µg/mL, the survival rate of ML was less than 50%. Furthermore, when treated with NPP at concentrations of 12.5, 25, 50, 100, and 200 µg/mL for 24 h, the transcription levels of the TsTPP gene decreased by 19%, 23%, 30%, 38% (all p < 0.05), and 57% (p < 0.01), respectively; after 36 h of treatment, the transcription levels of TsTPP gene were reduced by 18% (p < 0.05), 23%, 35%, 39%, and 60% (all p < 0.01); after 48 h of treatment, the transcription levels of TsTPP gene decreased by 19%, 23%, 42%, 42%, and 65% (all p < 0.01), respectively. N-(phenylthio) phthalimide (NPP) exhibits significant lethal activity against T. spiralis by inhibiting the trehalose synthesis pathway, providing basic data for future NPP as a new clinical drug target.

During a 2024 survey for actinosporeans at Tasik Telabak, Malaysia, three distinct aurantiactinomyxon types were identified from different specimens of the oligochaete Aulophorus sp. and characterized through both morphological and molecular analyses. Morphometric measurements revealed notable differences among the three types, particularly in the length of their caudal processes, despite all three types being found in the same host species. Analysis of the 18S ribosomal DNA (rDNA) of these aurantiactinomyxon types showed no close matches to existing myxozoan sequences in GenBank. Phylogenetic analyses placed all three types within a myxobolid clade dominated by Henneguya spp.; aurantiactinomyxon type 1 formed a subclade with Myxobolus miyairii from Silurus asotus, while aurantiactinomyxon types 2 and 3 clustered at the base of a larger subclade comprising mainly Henneguya species that infect Ictalurus punctatus and related catfish species. This study represents the second report of myxosporean infections in Malaysian oligochaete communities.

Blood flukes of the genus Cardicola (Digenea: Aporocotylidae) are well-documented health threats to farmed and ranched bluefin tuna (BFT), where infections can result in substantial economic losses. While recombinase polymerase assays (RPA) have been developed for two species of Cardicola, the high degree of sequence conservation across all three species reported from BFT requires an alternative method to enable confident species-level discrimination. The aims of this study were to design, optimise and validate loop-mediated isothermal amplification (LAMP) for the detection of the three Cardicola species which infect BFT, and apply the assays to Pacific bluefin tuna (PBT) cultured in Japan. Species-specific LAMP assays were successfully designed, optimised and validated for C. forsteri, C. opisthorchis and C. orientalis targeting ITS-2 ribosomal DNA. In addition, these assays were applied to DNA from three sample types targeting sites of significant pathology, the heart, gill necropsy and gill biopsy of 15 PBT. Results were compared to outputs of quantitative polymerase chain reactions (qPCR), the gold standard for blood fluke detection in ranched Australian SBT, where LAMP assays were shown to be more sensitive. This research constitutes the first development and application of species-specific LAMP diagnostics for the three described Cardicola species infecting BFT, providing a rapid, cost-effective and accessible alternative to standard molecular diagnostics, informing management and treatment decisions.

Blastocystis is one of the most prevalent gastrointestinal parasites in humans. This protist has been detected in water from various sources, with the highest prevalence in wastewater. In a previous study, we demonstrated that Blastocystis can adhere to and maintain viability within aquatic biofilms in wastewater. Therefore, the objective of this study was to evaluate wastewater contamination by viable Blastocystis at four sites along an urban stream (Buenos Aires, Argentina) using aquatic biofilms. Two sampling strategies were employed: retrieval from fallen and submerged plant leaves (natural surfaces) and from submerged glass slides (artificial surfaces) at three time points. Biofilms were cultured in Jones' medium to assess parasite viability, and Blastocystis subtypes were identified by PCR and sequencing of the SSU rRNA gene. Contamination levels at each site were evaluated 15 days after the glass slides were immersed, and the results were expressed as cysts per milligram of biofilm. Biofilms containing viable parasites were more frequently detected at sites with higher population density. The loads ranged from 1 to 61 cysts per milligram of biofilm, with remarkably higher values at the sites with most significant human impact. Molecular analyses revealed the presence of subtypes ST1 (allele 4), ST4 (allele 42), and ST8 (allele 21 and allele 95), with ST1 and ST8 being the most prevalent across sampling periods. The two sampling strategies proved useful, although their applicability depends on the specific objective of the study. This study provides the first evidence supporting the use of aquatic biofilms as an effective tool for monitoring viable Blastocystis in wastewater and for assessing spatial and temporal contamination patterns.

Yellowtail Kingfish (YTK) (Seriola lalandi) is widely distributed throughout the Indo-Pacific and is a significant recreational and commercial species in Australia. In New South Wales (NSW), a post-cooking mushy flesh condition has been reported, though its cause remains unconfirmed. This study combined morpho-molecular diagnostics with a citizen-science mushy fish reporting platform to identify associated parasites and to map the spatio-temporal distribution of mushy flesh and infections in NSW. Unicapsula seriolae was detected in 38.9% (95% exact CI: 23.1-56.5%) of YTK donated by recreational anglers and charter operators (n = 36) (2024-2025) and confirmed by myxospore morphology and SSU and LSU rDNA sequencing. Most infected fish (71.4%) did not exhibit the phenotype, although mushy individuals were significantly more likely to be infected than non-mushy fish. Infection occurred year-round and across bioregions, whereas the phenotype was restricted to warmer months, yet asymptomatic infections were also present. Collectively, these findings extend the records of U. seriolae-YTK infections on Australia's east coast beyond the initial reports off QLD and indicate that mushy flesh alone is an unreliable indicator of U. seriolae infection. Infection and sea surface temperature alone were insufficient to drive phenotypic expression, with host-environment-parasite interactions likely modulating its manifestation. Citizen-science data (n = 48) revealed widespread, temporally variable phenotypic occurrence and documented socio-economic impacts on anglers and consumers. The parasite's effects on flesh quality may influence angler and consumer perception and marketability; however, the weak concordance between infection and phenotype indicates that visual assessment alone is likely to underestimate infection, allowing infected fish to enter the supply chain, undetected. Although U. seriolae is not considered a significant zoonotic parasite, in Japan, reports of foodborne illness associated with U. seriolae in other Seriola species suggest these findings warrant consideration from a food safety perspective. The results highlight that U. seriolae is present in YTK from the Manning and Hawkesbury shelf bioregions in NSW, but the expression of mushy flesh is conditional rather than a direct, inevitable consequence of infection.

Cystic echinococcosis (CE) is a significant zoonotic infection caused by the larval stage of Echinococcus granulosus sensu lato, and it is a serious public health problem in Türkiye. The study aims to determine the current prevalence of bovine CE in Erzurum, to identify its molecular genotypes, and to analyze long-term trends in infection prevalence. A total of 1,241 slaughtered cattle were examined, and the CE prevalence was 24.09% (299/1,241). The lungs were the most frequently infected organ, with a rate of 93.31% (p < 0.05). The study revealed that the highest infection rates were observed in cattle over 5 years of age and in females (p < 0.05). The vast majority of infected cattle exhibited single cysts (73.24%) (p < 0.05), with cyst diameters ranging from 1 to 15 cm. Medium-sized cysts (2-5 cm) were predominant in 73.06% of cases (p < 0.05). Microscopic examinations revealed that the fertility rate of cysts was 9.25%. Cyst fertility significantly increased with cyst size (p < 0.05). A molecular analysis of 105 selected cyst samples targeting the 12 S rRNA and COI gene regions identified all isolates as E. granulosus sensu stricto. The epidemiological data from the last thirty years demonstrate a decrease in the prevalence rate of approximately 10% per decade in the region, with a decline from 46.4% to 24.09%. Consequently, in order to maintain the current downward trend in prevalence observed in CE endemic regions and to minimize public health risks, it is crucial to implement comprehensive control programs based on a 'One Health' approach and to continue periodic epidemiological studies to monitor the parasite's status in hosts.

Malaria remains a major global health threat, particularly in endemic regions of sub-Saharan Africa. In 2024 alone, an estimated 282 million cases were reported worldwide, with approximately 610,000 associated deaths. Beyond the high disease burden, mixed-species Plasmodium infections represent an additional clinical challenge, as they are frequently underdiagnosed and may significantly complicate both diagnostic accuracy and therapeutic management even in non-endemic countries. We report the case of a 67-year-old man from Guinea who presented to the Emergency Department with high-grade fever and asthenia shortly after returning to Italy; initial microbiological investigations were negative, despite markedly elevated inflammatory markers were diagnosed. The patient reported a previous untreated malaria episode approximately six months earlier, raising suspicion of a novel Plasmodium infection. A comprehensive malaria diagnostic workflow was promptly initiated, integrating rapid immunoenzymatic testing (IE), microscopy, and molecular assays. The IE test suggested Plasmodium falciparum or a mixed infection as result, while loop-mediated isothermal amplification (LAMP-PCR) confirmed the presence of Plasmodium spp DNA. Microscopic examination of stained blood smears revealed typical P. falciparum ring forms and suspected P. ovale trophozoites; so, species-specific real-time PCR confirmed a mixed infection with P. falciparum and P. ovale, supporting prompt antimalarial treatment. Final molecular confirmation was achieved by Sanger sequencing, which identified the Plasmodium ovalecurtisi as co-aetiological agent of disease. The rapid and integrated diagnostic approach enabled timely initiation of appropriate antimalarial therapy with artemether-lumefantrine and primaquine, leading to complete clinical recovery and gene sequencing allowed a correct species confirmation and epidemiological characterization. This case highlights the importance of combining microscopic analysis with advanced molecular diagnosis for an accurate identification of mixed Plasmodium infections even in country and contributes novel epidemiological data through molecular characterization of P. ovalecurtisi in an imported case.

Cases of adventitious feeding on humans by non-hematophagous insects hold significant medical relevance, yet their occurrence during active body art procedures remains undocumented. This case report describes a persistent instance of opportunistic exsudatophagy by the house fly, Musca domestica Linnaeus, 1758 (Diptera: Muscidae), during a tattooing session on a healthy 32-year-old male client in Germany. Over a four-hour timeframe, four adult house flies entered the ground-floor studio through doors and windows opened for summer cross-ventilation. The insects exhibited highly persistent landing and feeding attempts directly on the freshly wounded tissue of the right lower leg, actively drinking a matrix of blood, plasma, and tattoo ink. Attraction intensified significantly once large-scale shading expanded the exuding epidermal surface area. Effective mechanical repulsion was heavily restricted by the client's enforced immobility required to ensure precise tattoo application. Following a routine aftercare protocol, the lesion healed completely without secondary bacterial infection. This report highlights a neglected hygienic hazard where the saprophagous and coprophilic ecology of synanthropic filth flies poses a mechanical vector risk for dermal pathogen inoculation, emphasizing the necessity of simple vector control strategies in tattooing facilities.

Praziquantel is a widely used anthelmintic in both human and veterinary medicine, but low palatability of the commonly used compound, which is a mixture (M) of two isomers (D and L forms), may result in rejection of oral treatments. Purification of the D-isomer of praziquantel allows a comparative evaluation of both palatability and efficacy of the isomers. We showed that rainbow trout (Oncorhynchus mykiss) preferred feed containing D-PZQ feed (60% acceptance) compared to M-PZQ feed (20% acceptance) (inclusion of 400 g PZQ/kg feed). Subsequent efficacy studies using eyefluke Diplostomum pseudospathaceum cercariae (in vitro) and experimentally infected rainbow trout carrying metacercariae (in vivo) demonstrated a strong antiparasitic effect of both PZQ-forms. In vitro assays documented that both M-PZQ and D-PZQ induced rapid and concentration-dependent mortality in cercaria (100% within 4 h in 0.125 mg PZQ/L), but with a significantly higher killing rate of of D-PZQ. In vivo assays (17 days feeding medicated feed) showed significant efficacy of both D-PZQ and M-PZQ. A slightly (non-significant) faster effect of D-PZQ may partly be ascribed to a combination of higher consumption of this isomer (due to its higher palatability) and higher efficacy. Cataract formation occurred in control fish but was prevented by both drug types indicating that metabolic and/or mechanical activities of live parasites are associated with cataract induction. This rainbow trout/eyefluke infection may serve as a model for non-piscine host-parasite systems as well.

Objective Sparganosis is a rare parasitic disease caused by the larvae of Spirometra mansoni, with pediatric cases being particularly uncommon and prone to misdiagnosis. This study aims to analyze the clinical features, diagnostic strategies, and treatment responses in pediatric sparganosis, and to investigate its global epidemiological trends and diagnostic and therapeutic advancements through a systematic review. Methods We conducted a retrospective analysis of 5 confirmed pediatric sparganosis cases admitted to our center between 2014 and 2023. A systematic search of the PubMed database (1980-2025) for relevant literature was performed, leading to the screening and data extraction of 135 global case reports. Descriptive statistics, correlation analysis, and trend visualization were conducted using R software (version 4.5.1). Results Among the 5 children in our center, the median age was 7 years, and all were from high-parasitic-endemicity areas in Yunnan Province, China. All cases (100%) presented with marked eosinophilia (median value 4.42 × 10[9]/L). Neurological involvement was predominant (60%, 3/5), manifesting as epilepsy, myelitis, etc. All patients received praziquantel treatment, with four also receiving concomitant corticosteroids, and all showed symptom improvement. Analysis of global cases revealed a geographic clustering in East Asia (primarily China, South Korea, Japan, and Thailand), with the central nervous system and subcutaneous tissue being the two major infection sites. Diagnosis relied mainly on MRI ("tunnel sign"), serological testing, and metagenomic Next-Generation Sequencing (mNGS). The highest improvement rate (92.2%) was observed with combination therapy (surgery plus medication), which yielded comparably high improvement rates to single-modality treatments. Conclusion Pediatric sparganosis predominantly involves the nervous system. Diagnosis requires integrating epidemiological history, imaging, and molecular testing. Treatment should be individualized, with a recommended regimen of surgery combined with long-term, weight-based praziquantel therapy, supplemented by corticosteroids when necessary. Strengthening public health education remains the fundamental measure for preventing this disease.

Ticks are important vectors of pathogens with substantial medical and veterinary relevance, a threat that may be further increased with the spread of invasive tick species. Reliable tick identification is relevant for accurate diagnostic elucidation of transmission pathways, and the development of targeted surveillance strategies and risk assessments. A standardized, rapid, and cost-effective identification workflow that can be implemented across laboratories is needed. Previous studies have shown that MALDI-TOF MS is a promising, objective, and economical approach for tick identification. However, the methodology remains insufficiently standardized and is not yet routinely applied for vector identification. The aim of this study was therefore to optimize MALDI-TOF MS-based tick identification for routine use in diagnostic laboratories. A simplified and standardized protocol for protein extraction from whole tick legs via brief incubation in 70% formic acid is presented. This protocol provides a robust basis for the development of a curated MALDI Biotyper® reference library, enabling reliable, expert-independent tick identification and supporting epidemiological surveillance of vectors and vector-borne pathogens.