In this study, the anthelmintic potential of the ethanolic and aqueous extracts of Moringa oleifera leaves and Embelia ribes seeds was evaluated against Haemonchus contortus by both in vitro (egg hatch assay, EHA; larval development assay, LDA; and adult mortality assay, AMA) and in vivo assays (faecal egg count reduction test, FECRT). Ethanolic extract of M. oleifera (MOEE) exhibited maximum inhibitory effects on egg hatching (87.77%) and larval development (86.97%) along with complete adult mortality (100% after 12 hours of incubation) @ 16.0 mg/mL and 32.0 mg/mL, respectively. However, ethanolic extract of E. ribes (EREE) @ 16.0 mg/mL concentration exhibited poor ovicidal (22.69%) but moderate larvicidal (77.90%) and adulticidal (80% mortality after 12 hours of incubation) effects. In EHA, MOEE showed a moderate IC50 of 5.721 mg/mL, while EREE failed to achieve 50% inhibition at tested concentrations (1–32 mg/mL). For the LDA, both MOEE and EREE exhibited moderate IC50 values of 4.131 mg/mL and 2.882 mg/mL, respectively. The aqueous extract of M. oleifera (MOAE) exhibited highest inhibitory effect on egg hatching (89.12%), larval development (89.94%), and also caused significant mortality (100%) of adult H. contortus in comparison to the standard drugs and other plant extracts evaluated in this study. In contrast, ERAE showed no significant effect on egg hatching or adult worm viability but notably inhibited larval development (86.97%). Both of the plant extracts contained phytochemicals like phenolic compounds, tannins, flavonoids, and alkaloids with anthelmintic properties. Further, in vivo trial using MOAE @ 150 mg/kg BW exhibited 72.42% FEC reduction on day 10 post-treatment as compared to fenbendazole (70.92%) and levamisole (89.01%) treated groups. The MOAE appeared effective in combating the resistant H. contortus worms. This study formed a base for further in vivo evaluation of anthelmintic efficacy of the mixture of Moringa and Embelia extracts and to standardise the optimum concentration and dose for its field applicability against GI parasitism in animals.

The Trematoda family Didymozoidae comprises over 250 species that typically use marine fishes as their definitive hosts. Species identification of adult didymozoids based solely on morphology is often problematic owing to the difficulty of observing their detailed internal organs. Although molecular analyses are generally essential for trematode taxonomy, reference sequences (e.g. 28S rDNA, ITS, and COI) for morphologically identified adult didymozoids remain scarce. In this study, adult didymozoids were collected from five marine fish species in Japan between 2018 and 2025. Morphological observations revealed four species, including Annulocystis carnata n. sp. from Euthynnus affinis. The new species is distinguished from the other members of the genus Annulocystis by its robust forebody, subglobular oral sucker, and oval testes. Three known species, Annulocystis auxis (from Auxis thazard), Gonapodasmius squamatus (from Mola mola), and Nematobothrium filiforme (from Scomber spp.), are redescribed and linked to novel molecular markers. The 28S rDNA sequence of Annulocystis carnata n. sp. was identical to that of a previously reported Annulocystis cf. auxis from Australia, suggesting that they are conspecific. Our molecular analyses revealed that some metacercariae reported from cnidarian jellyfishes in a past study were conspecific with N. filiforme. These results suggest the critical need for molecular markers in didymozoid identification. Our 28S rDNA-based phylogenetic analysis indicates that the currently accepted macrotaxonomy of the family may require revision for some genera and subfamilies.

Neodiplostomum Railliet, 1919, is a cosmopolitan genus that parasitises mainly the intestines of birds of prey, and occasionally those of passerines, cuculiforms, and mammals. Adults of three undescribed species of Neodiplostomum were obtained from five bird species from three families (Accipitridae, Cuculidae, and Tyrannidae) in a single locality. Four molecular markers, namely, the small (SSU) and large (LSU) subunits and internal transcribed spacers (ITS1 and ITS2 plus 5.8S) from nuclear ribosomal DNA, and cytochrome c oxidase subunit 1 (cox 1) from mitochondrial DNA, were sequenced from specimens of the three species. The newly obtained sequences were aligned with other sequences of Neodiplostomum spp. downloaded from GenBank. Maximum likelihood and Bayesian analyses based on each molecular marker revealed that the specimens of the three undescribed species of Neodiplostomum formed three independent lineages with high values of genetic divergence; thus, they are recognised herein as three new species: Neodiplostomum acatlotlii n. sp., N. papaloapanensis n. sp., and N. leoni n. sp. The new species showed morphometric differences in the body size, prosoma, opisthosoma, suckers, holdfast organ, testes, ovaries, and eggs. Our study also revealed that the three new species are distributed sympatrically in the Veracruzan biogeographical province in the Neotropical region of Mexico and use different ways of transmission to complete their life cycles.

The structuring of parasite communities in small mammals depends on both host-intrinsic factors and environmental conditions. We investigated the helminth fauna of the caviomorph rodent Thrichomys laurentius, comparing populations from two contrasting phytophysiognomies in the Caatinga biome: open shrubland in Carrasco area and seasonal deciduous forest in Mata Seca area. In this context, the influence of biotic predictors (host sex and host body weight) and abiotic predictors (phytogeographic subunit: Carrasco or Mata Seca) on parasite diversity (species richness, Shannon, and Simpson indices) was evaluated, as well as the effect of habitat type on the parasite community composition of this host species. Twenty-seven T. laurentius specimens (Carrasco: n = 15; Mata Seca: n = 12) were examined for helminths. A total of seven helminth species were recovered (Heligmostrongylus almeidai, Paraheligmonella interrogans, Helminthoxys freitasi, Stilestrongylus freitasi, Stilestrongylus sp., Raillietina sp., and Aspidodera raillieti). Heligmostrongylus almeidai was the dominant species in both phytophysiognomies, sharing dominance with P. interrogans in the Mata Seca. Overall helminth infection was significantly higher in Mata Seca (100%) compared with Carrasco (53%). The phytogeographic subunit was the strongest predictor for diversity indices, followed by host sex, which influenced parasite richness, and host body weight, which influenced Shannon and Simpson diversity indices. Although Mata Seca showed higher parasite abundance, species diversity was higher in Carrasco due to lower dominance. Local physiognomic variations in the Caatinga impose environmental filters that govern geohelminth success. Additionally, this study reports the first record of Aspidodera raillieti in T. laurentius.

Gastrointestinal parasitic infections adversely affect the health and well-being of animals worldwide. The limitations of the efficacy of pharmaceutical anthelmintics underscore the importance of finding alternative approaches to reduce the impact of parasitism on hosts. Investigating the anthelmintic properties of bioactive plants could be one way to contribute to this effort. The present study evaluates in vitro the activity of aqueous and ethanolic extracts of Setaria barbata on the four free-living life stages of Ancylostoma caninum, the primary gastrointestinal nematode of dogs. After preparing the extracts, phytochemical tests were conducted to identify secondary metabolites based on colour and precipitation reactions specific to each metabolite present in the extracts. Serial dilution of the extract allowed us to obtain five test concentrations, which were used for egg embryonation, egg hatching, and larval mortality assays. Fresh eggs, embryonated eggs, and L1 and L2 larvae of A. caninum from a naturally infected dog were incubated for 24 hours with 5 concentrations of Setaria barbata extracts, while the embryonated eggs were incubated for 6 hours. 0.5% DMSO and distilled water were used as negative controls. At the end of this period, the number of non-embryonated eggs, unhatched eggs, and dead L1 and L2 larvae was recorded. Flavonoids, phenolic compounds, and alkaloids were identified in the three extracts of S. barbata. The presence of triterpenes and steroids, however, varied depending on the solvent used for extraction. The extracts proved more effective on embryonated eggs than on fresh eggs; the aqueous macerated extract thus resulted in 100% inhibition of egg hatching at a concentration of 0.625 mg/mL, with the lowest IC₅₀ (0.31 mg/mL). L2-stage larvae were more sensitive to the various extract concentrations than L1-stage larvae. The ethanolic extract caused 100% mortality in L2-stage larvae, with an LC₅₀ of 0.48 mg/mL. Extracts of Setaria barbata contain compounds with bio-ovicidal and bio-larvicidal properties. These extracts could be sprayed in dog kennels and in the wild as a disinfectant to eliminate the pre-infectious forms of gastrointestinal nematodes. Further studies are needed to evaluate the efficacy and safety of these extracts in a complete biological system.

Studies of bat helminth communities are relatively rare compared with those of other vertebrate groups. Additionally, the relationships between host attributes and environmental factors and the presence and abundance of helminths in bats have seldom been examined. In this study, we evaluated how host traits and environmental conditions may influence the prevalence and abundance of helminths in two cave-dwelling populations of the broad-tailed bat Nyctinomops laticaudatus in the state of Yucatán, Mexico. From January 2024 to May 2025, specimens of N. laticaudatus were captured from Calcehtok and Homún and examined for helminths. The helminth fauna recorded in N. laticaudatus consisted of 17 taxa, seven of which exhibited a prevalence higher than 10% and were analysed using generalised linear models and generalised linear mixed models: Brachylecithum rileyi, Limatulum nanum, Ochoterenatrema sp., Urotrema spp., Vampirolepis spp., Aonchotheca pulchra, and Anoplostrongylus allami. Prevalence and abundance were associated with the host traits and environmental factors analysed. Females had a higher probability of infection with helminths such as B. rileyi. Prevalence tended to differ between juveniles and adults for Urotrema spp. Non-reproductive bats and individuals from Calcehtok exhibited higher prevalences of parasites such as Vampirolepis spp. and L. nanum, respectively. Ochoterenatrema sp. and Vampirolepis spp. exhibited lower prevalence and abundance during months with reduced rainfall and higher temperatures, respectively. This study provides new information on the helminths associated with molossid bats and highlights the influence of host attributes and environmental conditions on helminth community structure.

Hemicriconemoides spp. are sheathoid, root-ectoparasitic nematodes with complex taxonomy due to overlapping morphology and cryptic diversity. In this study, an integrative taxonomic approach combining morphology and molecular data supports the characterization of Hemicriconemoides vietnamensis sp. n. recovered from the rhizosphere of roses in Vietnam. The new species females are distinguished by a slightly ventrally curved body with a closely fitting sheath lacking anastomoses, a lip region with two annuli and a protruding first annulus, a long (62–68 μm) mostly straight stylet with rounded, posteriorly sloping knobs, and an excretory pore located 5–7 annuli posterior to the pharyngeal base. The vulva is slit-like without vulval flaps, the vagina is oblique, and the tail is bluntly to pointed conical with a rounded terminus. Males were not found. Morphologically, the new species is most similar to H. litchi, H. rotundoides, and H. strictathecatus, but can be differentiated by a combination of morphological and molecular characterizations. Molecular analyses of ITS rDNA, D2–D3 expansion segments of 28S rDNA, and COI mtDNA consistently support the distinct status of H. vietnamensis sp. n. Bayesian phylogenetic analyses place the new species in a well-supported clade with H. phoenicis and H. strictathecatus, forming a distinct lineage across all markers. Species delimitation analyses (bPTP, GMYC, and ASAP) consistently recover Hemicriconemoides vietnamensis sp. n. as an independent species. These results provide clear evidence for the establishment of Hemicriconemoides vietnamensis sp. n. and expand the documented diversity of Hemicriconemoides in Vietnam, where molecular data for the genus remain limited.

This study investigated the population dynamics and reproductive biology of the acanthocephalan Octospiniferoides incognita in the intestine of the herbivorous fish Rhytiodus microlepis throughout the seasonal water level fluctuations of a floodplain lake in Amazonas State, Brazil. Between July 2022 and November 2023, hosts were sampled during the rising water, high water, receding water, and low water phases. The results show that the water level fluctuations significantly influence infection levels. Prevalence was 73% during rising water, reached 100% during high water, and remained high during receding water (89%), whereas no parasites were recorded during low water. Abundance and mean intensity were higher during high water and receding water. The population of O. incognita showed sexual dimorphism, with females attaining larger body sizes than males, and both sexes occurring preferentially in the middle portion of the intestine. Three reproductive morphotypes were recognized for each sex. Copulatory activity began during rising water, whereas the highest egg production was concentrated during high water and early receding water. Size at first sexual maturity (L50) was estimated at 0.12 cm, indicating an early maturation strategy. These findings clearly show that the infection dynamics and reproductive processes of O. incognita are closely synchronized with the flood pulse, which regulates host availability and promotes the success of trophic transmission in the Amazonian floodplain ecosystem.

Phenotypic plasticity allows parasites to modify their morphology in response to the ecological and physiological conditions induced by their hosts. In digeneans with complex life cycles and multiple definitive hosts, host-induced variation may complicate species delimitation and taxonomic diagnoses. In this study, we characterised intraspecific morphological variation in Mesorchis uruguayensis (Digenea: Echinochasmidae), a trematode that parasitises fish-eating birds, using an integrative morphological approach. Thirty-eight adult specimens previously collected from five definitive host species in Mexico and molecularly identified as M. uruguayensis were examined. Scanning electron microscopy (SEM) was used to assess qualitative diagnostic traits, and principal component analysis (PCA) based on 19 linear measurements was used to explore morphometric variation among host-associated groups. All specimens retained the characteristic head collar with 22 spines, 10 small spines along the anterior margin of the oral sucker, and a consistent general pattern of tegumental spines. The first two principal components explained 80.9% of the total morphometric variation. However, the host-associated group overlapped substantially, with some differences. Body size, the dimensions of reproductive organs, and the ventral sucker contributed most strongly to this variation. Because all specimens were molecularly conspecific, these differences represent intraspecific morphological variation rather than species-level divergence. Host identity constitutes a biologically plausible source of this variation through differences in intestinal physiology, resource availability, and other host-associated conditions; however, geographic origin, parasite maturity, infection intensity, and individual host condition may also contribute. Our findings emphasise the value of combining stable qualitative characters, morphometric analyses, and molecular evidence to interpret morphological variability and avoid taxonomic misidentification in digeneans.

Translocations of rhinoceros in the 1990s provided an opportunity to investigate the vectors of filaria in Nairobi National Park and Solio Ranch in Kenya. These are high-altitude sanctuaries where rhinos have ulcerative sores attributed to the filarial nematode Stephanofilaira dinniki. Rhinomusca dutoiti, the biting fly closely associated with rhino, did not harbour S. dinniki, but often harboured a small and/or a large form of Habronema. Rhinomusca dutoiti from Kenya was nearly identical to R. dutoiti from South Africa based on gross morphology and a 798 base pair sequence from the 3′ end of the Cytochrome Oxidase I gene. Habronema parasites occurred alone or in combination and were smaller than larval forms of H. muscae, the well-known parasite of equines. At Nairobi, prevalence of infections in the proboscis was high at 55%, with mature infections already present in young flies. This finding implied infection during larval development rather than acquisition as an adult fly. Infection patterns were similar in flies trapped or captured by hand from rhinos at Solio Ranch. Habronema was also found in a few Haematobosca spp., at much lower prevalence. Parasites were measured for several body landmarks and were photographed through light microscopy and scanning electron microscopy. These data now provide a foundation for further studies of the biology of Rhinomusca and its relationship with filarial nematodes in wildlife.

We surveyed trematode parasites infecting 227 gulls from southern New Zealand, comprising 113 red-billed gulls (Chroicocephalus scopulinus) and 114 black-backed gulls (Larus dominicanus), using morphological and molecular approaches. Fifteen trematode species were recovered, including six species new to science: Mesorchis carsonae n. sp., Acanthoparyphium argillamartelliarum n. sp., Himasthla davisi n. sp., Gymnophalloides hutsonae n. sp., Levinseniella trisepsilon n. sp., and Microphallus lagruei n. sp. Nine species occurred in both gull hosts; the remaining six were recorded from only black-backed gulls. New genetic data were generated for 12 species, including all new species, enabling confirmation of species identities and links to larval stages in intermediate hosts, thereby resolving or advancing knowledge of several life cycles. Several parasites represent new host or geographic records, including the first named species of Diplostomum, Gigantobilharzia, Levinseniella, Gymnophalloides, Microphallus and Mesorchis reported from New Zealand. Parasite burdens were generally low, but species richness was high, underscoring gulls as important definitive hosts supporting substantial hidden biodiversity. This study provides the first comprehensive baseline survey of trematode communities in New Zealand gulls and highlights that parasite diversity remains underestimated even in comparatively well-studied hosts. These findings reinforce the importance of incorporating parasites into biodiversity inventories and conservation frameworks.

This study was conducted to determine the in vivo anthelmintic efficacy and clinical safety of crude ethanolic extract (CEE) obtained from Artemisia absinthium in naturally infected Awassi breed sheep with gastrointestinal nematodes (GIN). A total of 18 non-pregnant sheep were divided into 3 groups of 6 animals each: CEE (1.0 g/kg), albendazole (5 mg/kg), and negative control (untreated). Changes in egg counts (EPG) were quantitatively monitored in faecal samples collected before treatment (day 0) and after treatment (days 7 and 15). The research findings showed that A. absinthium CEE significantly reduced the parasite load over time, with the highest efficacy achieved on day 15 (p < 0.001), demonstrating statistically significant efficacy in suppressing the fecundity of GIN agents. According to the faecal egg count reduction test (FECRT), therapeutic success was observed in 76.77% of the CEE group and 100% of the albendazole group. The performance demonstrated by CEE confirmed the potential of plant secondary metabolites to reduce dependence on synthetic drugs, and the absence of any adverse effects during the trial period demonstrated the plant’s high biosafety profile. Consequently, A. absinthium offers a powerful, economical, and residue-free biotherapeutic alternative to synthetic anthelmintics in sustainable and organic livestock models, countering global anthelmintic resistance. To the best of our knowledge, this study represents the first investigation in Türkiye evaluating the anthelmintic efficacy of A. absinthium CEE in Awassi breed sheep naturally infected with GIN, providing the first regional efficacy data from Şanlıurfa.

In the current study, we used an immunotherapeutic approach in female Swiss albino mice inoculated intraperitoneally with viable protoscoleces. We investigated the potential immunomodulatory effect and protective properties of IL-17A at 125 pg/mL on the development of secondary experimental echinococcosis. Swiss mice were inoculated intraperitoneally (i.p.) with 200 μL of a suspension containing 2000 viable protoscoleces to establish the infection model. IL-17A was administered intravenously two weeks after cystic echinococcosis (CE) induction. Cyst development and hepatic damage were macroscopically and histologically analysed. The hepatic expression of iNOS, TNF-α, NF-κB, CD68, and vimentin was examined. We observed that in vivo IL-17A treatment at 125 pg/mL reduced metacestode growth by 93.8%. Interestingly, treatment at this dose also led to decreased splenomegaly and improved liver tissue architecture. This was associated with a marked reduction in the fibrotic response, as evidenced by decreased collagen and reticulin fibre deposition. Moreover, IL-17A treatment resulted in reduced expression of key inflammatory and fibrotic markers, including iNOS, TNF-α, NF-κB, CD68, and vimentin, in the liver parenchyma. Our findings highlight the antihydatid and immunoprotective effects of IL-17A and support its potential therapeutic role in managing CE.

Short-term thermal stress during heatwaves can potentially affect the transmission of parasites by increasing mortality of infective stages. Using the critical thermal maximum approach (CTmax), commonly used in free-living species, we exposed trematode cercariae of three marine species (Cryptocotyle lingua, Himasthla elongata, Renicola parvicaudatus) to a temperature ramping rate similar to realistic heatwave scenarios in the field and determined their acute temperature limits. The median CTmax values of all cercariae were much higher than the typical temperature extremes observed during heatwaves in the field. CTmax values slightly differed among the three trematode species and were higher than the thermal limit of their first intermediate host, the periwinkle Littorina littorea. This suggests that heatwave effects on trematode cercariae are likely not direct but more indirect, resulting from effects on the survival of their first intermediate host. Given its straightforward methodology and the potential for standardized comparisons of thermal limits among species and ecosystems, the CTmax approach holds promise for ecological research on the thermal biology of parasites.

A new spiral nematode species, Helicotylenchus laichauensis sp. n., is described from the rhizosphere of Panax vietnamensis var. fuscidiscus in northern Vietnam. The new species is characterised by a hemispherical lip region with 4–5 annuli, a lateral field with four incisures whose inner lines fuse distally into a distinct m-shaped pattern on the tail, and a well-developed stylet 24.5–26.7 μm long, with knobs flattened to slightly indented anteriorly and a conus forming 50.1–53.5% of the total stylet length. The spermatheca is round and filled with sperm, and males are common. The tail is conical, dorsally convex, tapering to a prominent ventral projection bearing a variable mucro; phasmids are located 3–7 annuli anterior to the anus. Based particularly on similarities in the lip region structure, stylet morphology, lateral field pattern, phasmid position, and tail configuration, the H. laichauensis sp. n. is close to the known species namely H. microcephalus, H. craigi, H. oleae, and H. amplius. Phylogenetic analyses based on the D2–D3 expansion segments of 28S, partial 18S, and ITS rRNA sequences revealed that the newly generated sequences of the new species form a maximally supported clade with corresponding sequences of Helicotylenchus microcephalus in all constructed trees.

Accurate discrimination of Taenia solium (T. solium) from other Taenia species remains a challenge in parasitological diagnosis, particularly when based solely on morphological examination of eggs or degraded material. Molecular approaches have improved species identification and differentiation, but robust and species-specific targets suitable for developing diagnostic assays remain limited.

In this study, we aimed to identify and validate a species-specific molecular target to enable the discrimination between T. solium and other Taenia species. To this end, we performed comparative analysis of publicly available genomic sequences, revealing polymorphic sites unique to T. solium within a conserved locus; sequence alignments across Taenia species then guided primer design targeting this region. The resulting target was subsequently evaluated by quantitative PCR (qPCR) and showed consistent amplification across serial DNA dilutions, with detection down to approximately 10 pg of input DNA. No amplification was observed with genomic DNA from Taenia saginata or human samples, supporting the specificity of the selected target under the conditions tested.

Together, these results identify a robust molecular target for the specific detection of T. solium, providing a useful foundation for the development of molecular diagnostic tools which may support improved detection strategies for this medically significant helminth.

Haemonchus contortus is a highly pathogenic gastrointestinal nematode of domestic and wild ruminants whose epidemiological success is closely linked to high reproductive capacity. For female daily egg output, however, direct observations remain limited, and published estimates vary widely, ranging from fewer than 20 to 10,000–15,000 eggs per female per day. This study assessed in utero egg counts in female H. contortus recovered from experimentally infected lambs in order to evaluate their implications for fertility relative to values commonly cited in the literature. Archived female nematodes from a previously published experiment were used to conduct the study. In total, 147 female worms recovered from 14 sheep were examined. It was found that the average in utero egg count was 345 (SD = 145) eggs per H. contortus female. Regression analysis provided no clear evidence that the experimental design substantially influenced this parameter. In addition, female body length was positively associated with egg counts, though the relationship was not strictly linear. Overall, the present study supports moderate rather than high estimates of female daily egg output for H. contortus.

The trematode Maritrema gratiosum Nicoll, 1907 has a heteroxenous life cycle comprising a marine gastropod as its first intermediate host, a barnacle as its second intermediate host, and a bird as its definitive host. Globally, the species is widely distributed from subtropical waters of the South China Sea to arctic waters of the Barents Sea. In European waters (namely, western Europe and the Barents Sea), the complete life cycle of M. gratiosum is known. Elsewhere in its range, however, its life cycle remains incompletely understood. Clarifying its distribution and resolving unknown host associations are essential for advancing a more holistic view of host–parasite dynamics and their ecological role in intertidal ecosystems. In this study, M. gratiosum was detected using DNA-based assays in individuals of the barnacle Balanus glandula Darwin, 1854 that were collected from the rocky intertidal zone along the southern coast of British Columbia, Canada. This finding provides the first molecular evidence of this trematode species in the Pacific, complementing earlier records. The same molecular approach can be extended to identify region-specific hosts, such as the first intermediate (molluscan) host in the Pacific region, which remains unidentified.