Trematode Genus Research Articles

Gastrointestinal Parasitic Infections in Non-Human Primates at Gabon’s Primatology Center: Implications for Zoonotic Diseases

Parasites and infectious diseases pose significant threats to primate populations, especially in captive non-human primates (NHPs). This study aimed to assess the diversity and prevalence of intestinal parasites in NHPs at the CIRMF Primatology Center. A total of 97 fecal samples were analyzed using parasitological techniques, including sodium chloride flotation and modified Baermann sedimentation methods. An overall parasite prevalence of 93.81% (91/97) was observed. Sixteen groups of parasites with zoonotic potential were identified, comprising ten genera of nematodes (Trichuris, Enterobius, Hookworm, Trichostrongylus, Mammomonogamus, Spirure, Oesophagostomum, Schistosoma, Ascaris, and Strongyloides), three genera of protists (Eimeria, Balantioides coli/Buxtonella, and Entamoeba), one genus of cestodes (Hymenolepis), and two genera of trematodes (Dicrocoelium and Paramphistomum). High prevalences were noted for Oesophagostomum spp. (83.5%), Strongyloides spp. (52.58%), and Trichostrongylus spp. (50.52%). These findings underscore the potential role of the CIRMF Primatology Center in maintaining and facilitating the transmission of intestinal parasites with high zoonotic potential. The co-existence of human and NHP parasites in shared environments, such as zoos and research facilities, emphasizes the need for a holistic, One Health approach that addresses the interconnected health of humans, animals, and the environment. This study highlights the urgent need for collaborative strategies to mitigate the risks of zoonotic parasite transmission between NHPs and humans in captive settings.

Description and phylogenetic position of an unusual genus and species of allocreadiid trematode from a characid freshwater fish in South America

The worldwide distributed family Allocreadiidae Looss, 1902 is predominantly parasitic in freshwater fish and occasionally in amphibians and reptiles. It contains 15 genera, of which only Allocreadium, Creptotrema, Wallinia and Creptotrematina have been reported in Argentina. The records of Allocreadium species still require further verification. The discovery of an allocreadiid digenean parasitizing Pseudocorynopoma doriae led us to erect a new genus for which the name Martorellia n. gen is proposed. It is characterized by poorly developed ventrolateral muscular lobes and a transversely elongated ovary, with the most characteristic feature being the presence of vitelline follicles in two fields (one anterior at level of ventral sucker and one posterior to the testis), which are not confluent in the middle. The phylogenetic tree reconstructed with 28S rDNA sequences placed the new genus as the sister taxa of Creptotrematina. Although both genera are separated by a small genetic distance, it is comparable with those separating other allocreadiid genera. The discovery of Martorellia liciae gen et sp. nov. provides further evidence of the great diversity of Allocreadiidae in South America. The analysis of new samples from different South American hosts with an integrative taxonomic approach as the one used here allows a better understanding of the evolutionary history and biogeography of allocreadiids in the Americas.

Urogenital schistosomiasis, a leading cause of bladder squamous cell carcinoma

Abstract Introduction/Objective Schistosoma, genus of trematodes, commonly known as blood flukes are parasitic flatworms responsible for a group of infections in humans termed schistosomiasis and hundreds of millions of people are infected worldwide. In areas where Shistosoma is not endemic to the region most cases of bladder malignancy are of urothelial origin. In areas where Shistosoma is endemic bladder malignancy is more commonly squamous cell carcinoma. However, with increased travel, migration and tourism it is important to potentially consider Shistosoma infection in areas that are not typically considered endemic regions. Methods/Case Report A 62-year-old male presented to our hospital and revealed through an interpreter he has had 1.5 years hematuria with “pieces of meat or brown dirt”. Cystoscopy was performed and showed a large necrotic mass. CT scan showed an 8.2 cm exophytic bladder dome mass. TURBT was performed and a diagnosis of invasive high-grade urothelial carcinoma with squamous differentiation and with multiple Schistosoma organisms identified.The patient received Schistosoma treatment and was scheduled for radical cystectomy and ileal conduct. A few days before the scheduled procedure the patient presented to our emergency department with acute abdominal pain. He was subsequently diagnosed with progressive bladder cancer with new left malignant ureteral obstruction, which lead to hydronephrosis, DVT, abdominopelvic ascites and intraperitoneal bladder perforation. Abdominal fluid cytology showed atypical keratinizing squamous cells suspicous for malignancy and marked acute inflammation. Patient underwent cystectomy with ileal conduit and the specimen was sent to pathology for examination. Results (if a Case Study enter NA) NA Conclusion Gross examination showed an irregular perforated bladder. Microscopic examination showed a high grade urothelial carcinoma with squamous and sarcomatoid differentiation with detrusor muscle invasion. Additionally, Schistosoma organisms with a terminal spine and calcified eggs were identified. Some of the calcified egg material was engulfed by giant cells. The organisms are surrounded by a prominent eosinophilic infiltrate. Lymphovascular invasion is also noted. Schistosomiasis is associated with the development of squamous cell carcinoma of the bladder. It is important to maintain high oncological alertness for squamous cell carcinoma, particularly for patients arriving from endemic areas. Such as in our case, a patient from Somalia living in Kansas City.

Induction of hepatic fibrosis in mice with schistosomiasis by extracellular microRNA-30 derived from Schistosoma japonicum eggs.

Schistosomiasis is a zoonotic parasitic disorder induced by the infestation of schistosomes, a genus of trematodes. MicroRNAs (miRNAs) in egg-derived exosomes are crucial for modulating the host's immune responses and orchestrating the pathophysiological mechanisms. Although the exosomes secreted by S. japonicum contain abundant miRNAs, the specific roles of these miRNAs in the pathogenesis of schistosomiasis-induced hepatic fibrosis are yet to be comprehensively elucidated. The egg exosomes of S. japonicum secrete miRNA-30, a novel miRNA. In vitro, the effect of miRNA-30 was evaluated by transfecting HSCs with miRNA mimics. The target gene biosignature for miRNA-30 was predicted using the miRDB software. The effect of miRNA-30 in hepatic fibrosis was evaluated by either elevating its expression in healthy mice or by inhibiting its activity in infected mice by administration of recombinant adeno-associated virus serotype eight vectors expressing miRNA-30 or miRNA sponges. This novel miRNA can activate hepatic stellate cells (HSCs), the prinary effector cells of hepatic fibrosis, in vitro, i.e., it significantly increases the fibrogenic factors Col1(α1), Col3(α1), and α-SMA at both mRNA and protein levels. In addition, miRNA-30 may activate HSCs by targeting the host RORA gene. In addition, in vivo experiments were conducted by administering a recombinant adeno-associated viral vector to modulate the expression levels of miRNA-30. The overexpression of miRNA-30 in healthy mice significantly elevated the expression of Col1(α1), Col3(α1), and α-SMA at both the transcriptomic and proteomic scales. This overexpression was coupled with a pronounced augmentation in the hepatic hydroxyproline content. Conversely, the in vivo silencing of miRNA-30 in infected mice induced a considerable reduction in the size of hepatic granulomas and areas of collagen deposition. Hence, in vivo, modulation of miRNA-30 expression may play a pivotal role in ameliorating the severity of hepatic fibrosis in mice afflicted with S. japonica. The study results suggest that miRNA-30 may augment schistosomiasis-induced hepatic fibrosis through a probable interaction with the host RORA. Our study may improve the current theoretical framework regarding cross-species regulation by miRNAs of hepatic fibrosis in schistosomiasis.

Morphological and molecular data establishes Clinostomum dolichorchum n. sp. (Digenea: Clinostomidae) in the great blue heron Ardea herodias L. and American bullfrog Rana catesbeiana Shaw.

Clinostomum is a cosmopolitan genus of trematodes that infect piscivorous birds, freshwater molluscs, freshwater fish and amphibians. Herein, a novel species of Clinostomum is described based on morphological and molecular data from an adult in the oral cavity of the great blue heron Ardea herodias and metacercariae collected from the gills and skin of American bullfrog tadpoles Rana catesbeiana. The novel species shares similar qualitative and quantitative morphological features with a congener, Clinostomum marginatum, which has overlap in host and geographic distribution. The most notable morphological difference when compared to C. marginatum is the greater posterior testis length of the novel species. Molecular data resolved similarities with morphological comparisons to nominal species and supports the establishment of a novel species. Molecular data include partial small ribosomal subunit (18S rRNA gene), ribosomal internal transcribed spacer regions (ITS1, 5.8S rRNA gene, and ITS2), partial large ribosomal subunit (28S rRNA gene), cytochrome c oxidase subunit 1 gene (cox1), and nicotinamide adenine dinucleotide dehydrogenase subunit 1 gene (nad1) sequences. Phylogenetic analyses place the novel species in a sister clade to C. marginatum. Morphological and molecular data, combined with phylogenetic analyses support the establishment of Clinostomum dolichorchum n. sp.

Surface Ultrastructure and Molecular Studies of Clinostomum complanatum (Rudolphi, 1814) Braun, 1899 (Trematoda: Clinostomidae) Metacercariae in some Freshwater Fishes from Sulaimani Province, Iraq

Clinostomum is a trematode genus in the family Clinostomidae. The mentioned trematode species is parasitizing many fish species as intermediate hosts, while piscivorous birds and mammals are the main definitive hosts. Total of 58 metacercariae of Clinostomum larvae were dissected out from 25 infected fish specimens from eight different species from a number of water bodies of Sulaimani Province, Kurdistan Region, Iraq. In this investigation, 959 fish specimens were collected. These includes five cyprinid species: Capoeta trutta (Heckel), C. umbla (Heckel), Carasobarbus kosswigi (Ladiges), Cyprinion macrostomum Heckel and Garra rufa (Heckel), two leuciscid species: Alburnus sellal Heckel and Squalius lepidus Heckel and the mugilid fish Planiliza abu (Heckel). The prevalence of infection for each of these species was 1.8%, 5%, 20%, 1.5%, 1.7%, 4.9%, 4.9% and 1.3%, respectively. The morphology of the Clinostomum metacercariae was studied by using a compound microscope; their ultra-morphology was evaluated with a scanning electron microscopy (SEM); and the molecular analysis were performed by amplifying, sequencing and comparing the ITS1-5.8S-ITS2 and 28S rDNA gene loci from isolated Clinostomum metacercariae. The obtained sequences confirmed that all metacercariae of the Clinostomum, collected during the present study, represented with C. complanatum (Rudolphi, 1814) based on percent identity with sequences in the GenBank subject database. The molecular characterisations of the C. complanatum metacercariae in the present study were deposited in GenBank NCBI.

Fasciolosis Prevalence in Sacrificial Cattle of West Sumatra, Indonesia

Fasciola is a species of the trematode genus that can cause devastating parasitic disease, namely fasciolosis. Fasciola spp. infestation can affect ruminants, such as cows, buffaloes, goats, and sheep, resulting in economic losses to livestock owners. Ruminants are the definitive host for the Fasiola species. This cross-sectional research was conducted on Eid al-Adha/Qurban in July 2022 to determine the prevalence of fasciolosis in sacrificial cattle in Fifty Cities District, West Sumatra, Indonesia. A total of 106 samples of sacrificial cattle liver from the abattoir were investigated. Examination of the liver for the presence of Fasciola spp. was carried out by postmortem examination by removing the liver from the abdominal cavity immediately after slaughter. The livers of all sacrificial cattle were examined by systematic inspection, palpation, and incision for Fasciola spp. infestation. Necropsy results of samples indicated the prevalence of Fasciola spp. (36.79%), which was higher in female animals, compared to males. Based on age, the highest prevalence was at the age of > 4 years, (52.95%), followed by 2 years (39.62%) and 3 years (25.00%). Regarding the cattle breed, the highest prevalences were indicated in Pesisir cattle (47.61%), Simmental cattle (44.44%), Bali cattle (37.28%), Ongole cattle (20%), and Limousine cattle (14.28%). This study revealed that fasciolosis in sacrificial animals in Fifty Cities, West Sumatra, was influenced by gender. Therefore, the findings of this study suggest improving treatment protocol for the prevention of fasciolosis in sacrificial animals.

Morphological constraint obscures richness: a mitochondrial exploration of cryptic richness in Transversotrema (Trematoda: Transversotrematidae)

Species of Transversotrema Witenberg, 1944 (Transversotrematidae) occupy a unique ecological niche for the Trematoda, living externally under the scales of their teleost hosts. Previous studies of the genus have been impeded partly by limited variation in ribosomal DNA sequence data between closely related species and partly by a lack of morphometrically informative characters. Here, we assess richness of the tropical Indo-west Pacific species through parallel phylogenetic and morphometric analyses, generating cytochrome c oxidase subunit 1 mitochondrial sequence data and morphometric data for hologenophore specimens from Australia, French Polynesia, Japan and Palau. These analyses demonstrate that molecular data provide the only reliable basis for species identification; host distribution, and to a lesser extent morphology, are useful for identifying just a few species of Transversotrema. We infer that a combination of morphological simplicity and infection site constraint has led to the group displaying exceptionally low morphological diversification. Phylogenetic analyses of the mitochondrial data broadly support previous systematic interpretations based on ribosomal data, but also demonstrate the presence of several morphologically and ecologically cryptic species. Ten new species are described, eight from the Great Barrier Reef, Australia (Transversotrema chrysallis n. sp., Transversotrema daphnidis n. sp., Transversotrema enceladi n. sp., Transversotrema hyperionis n. sp., Transversotrema iapeti n. sp., Transversotrema rheae n. sp., Transversotrema tethyos n. sp., and Transversotrema titanis n. sp.) and two from off Japan (Transversotrema methones n. sp. and Transversotrema panos n. sp.). There are now 26 Transversotrema species known from Australian marine fishes, making it the richest trematode genus for the fauna.

Resurrection of Diplostomum numericum Niewiadomska, 1988 (Digenea, Diplostomatoidea: Diplostomidae) Based on Novel Molecular Data from the Type-Host

Diplostomum is a taxonomically problematic genus of trematodes, with many members harmful at the metacercarial stage to fish in aquaculture. We found metacercariae in the eye vitreous humor of the rudd, Scardinius erythrophthalmus; the bleak, Alburnus alburnus; and the roach, Rutilus rutilus, in Karelia (Russia) and identified them as Diplostomum numericum based on a combination of morphological and ecological data. Previously, D. numericum was thought to be conspecific with Diplostomum gavium. However, our phylogenetic analysis based on the sequences of the cox1 mitochondrial DNA gene did not support this hypothesis. We demonstrated the conspecificity of D. numericum with the larval trematode Diplostomum sp. Lineage 3 of Blasco-Costa et al. (2014) from the eye vitreous humor of European salmonid and cyprinid fishes. Therefore, we resurrected the species D. numericum and identified Diplostomum sp. Lineage 3 of Blasco-Costa et al. (2014) as D. numericum. Finally, we hypothesized that D. numericum was, in fact, conspecific with Diplostomum colymbi and that the latter was distinct from D. gavium sensu stricto.

Helminths in Invasive Raccoons (Procyon lotor) from Southwest Germany.

As hosts of numerous zoonotic pathogens, the role of raccoons needs to be considered in the One Health context. Raccoons progressively expand their range as invasive alien species in Europe. This study aimed to investigate the intestinal helminth fauna of raccoons in Baden-Wuerttemberg, Germany, as no such screening had ever been conducted there. In total, we obtained 102 animals from hunters in 2019 and 2020. Intestinal helminths were retrieved using the SSCT (segmented sedimentation and counting technique) and identified morphologically and by PCR-based Sanger sequencing. Fecal samples were assessed using the ELISA PetChekTM IP assay (IDEXX, Germany) and flotation technique. The artificial digestion method was employed for analyzing muscle tissue. We detected species of four nematode genera (Baylisascaris procyonis, Toxocara canis, Capillaria spp., and Trichuris spp.), three cestode genera (Atriotaenia cf. incisa/procyonis, Taenia martis, and Mesocestoides spp.), and three trematode genera (Isthmiophora hortensis/melis, Plagiorchis muris, and Brachylaima spp.). Echinococcus spp. and Trichinella spp. were not found. The invasive behavior and synanthropic habits of raccoons may increase the infection risk with these helminths in wildlife, domestic and zoo animals, and humans by serving as a connecting link. Therefore, it is crucial to initiate additional studies assessing these risks.

Molecular identification of Austrodiplostomum sp., an eye parasite among farmed tambaqui in Amazonia

Austrodiplostomum Szidat & Nani, 1951 is a genus of parasitic digenetic trematodes widely distributed in the Neotropical region. Infects a wide variety of species, families and requests for freshwater fish. We identify samples of Austrodiplostomum sp, based on metacercariae isolates from eyes of tambaqui (Colossoma macropomum), a fish of high commercial importance in Brazil, and widely consumed by the population of the northern region. The sequences obtained clustered with A. compactum. This is the first report of the occurrence of diplostomids in farmed tambaqui in Amazonia.

Scaphanocephalus spp. (Trematoda: Opisthorchiidae) in intermediate and definitive hosts of the Yucatán Peninsula, Mexico, with a re-description of Scaphanocephalus expansus

Summary Scaphanocephalus is a small trematode genus belonging to the family Opistorchiidae. The genus currently contains only three species associated with marine fish as intermediate hosts and fish-eating birds as definitive hosts. Here, specimens of Scaphanocephalus were collected from the Osprey, Pandion haliaetus, and the White mullet, Mugil curema in the Yucatán Peninsula, Mexico. We report for the first-time DNA sequences of adult specimens of Scaphanocephalus, particularly S. expansus, as well as a sequence of a different species sampled as metacercaria. Morphological comparisons of Scaphanocephalus expansus confirmed the identity of the adult specimens, with minor morphological variations; Scanning electron photomicrographs were included, and the species was re-described. Phylogenetic analysis based on 28S rDNA sequences showed that Scaphanocephalus is monophyletic within Opisthorchiidae and consists of three independent lineages. Sequences of adults are identical to those of S. expansus. Instead, the sequence of the metacercaria sampled from the mesentery of Mugil curema nested with specimens reported as Scaphanocephalus sp. from a labrid fish in the Mediterranean Sea, herein named it as Scaphanocephalus sp. 2.

Molecular link between the metacercariae and adults of four species Haematoloechus (Digenea: Plagiorchioidea), including scanning electron microscopy characterization.

Haematoloechus comprises a species-rich genus of trematodes commonly found in the lungs of amphibians across the globe. Of the approximately 70 species reported worldwide, Mexico stands out as a biodiversity hot spot for the genus, with 16 species described. In Los Tuxtlas tropical rainforest, in south-eastern Mexico, three species have been reported, two of them endemic of the frog species occurring in the area. Here, we added the record of a fourth species in the area, and partially elucidated the life cycle of these species using DNA sequence data from the 28S rRNA and the COI genes. Frogs, and a diverse array of species of aquatic insects were sampled in two lakes within the tropical rain forest. Adults and metacercariae of Haematoloechus were obtained. Both developmental stages were linked molecularly contributing to our understanding of their transmission pathways, and probable host-specificity patterns. Each of the four species of Haematoloechus were sampled in a particular species of anisopteran odonate, and therefore were considered as specialists towards their second intermediate host. We validated the presence of H. complexus in the region of Los Tuxtlas because their metacercariae, sampled in the anisopteran dragongly Agriogomphus tumens, matched with sequences of H. complexus from Nebraska and Illinois in USA. We further described the ultrastructure of the tegument of three of the four species of Haematoloechus, sampled from their definitive host (Rana vaillanti, Rana berlandieri), to contribute with characters that can be reliable for species delimitation of the genus.

Characterization of MicroRNA Cargo of Extracellular Vesicles Isolated From the Plasma of Schistosoma japonicum-Infected Mice.

Schistosoma is a genus of parasitic trematodes that undergoes complex migration in final hosts, finally developing into adult worms, which are responsible for egg production and disease dissemination. Recent studies documented the importance of extracellular vesicles (EVs) in the regulation of host-parasite interactions. Herein, we investigated the microRNA (miRNA) profiles of EVs isolated from host plasma at different stages of Schistosoma japonicum infection (lung stage: 3 days post-infection (dpi), and liver stages: 14 and 21 dpi) to identify miRNA cargo potentially involved in the pathogenesis and immune regulation of schistosomiasis. Characterization of the isolated plasma EVs revealed their diameter to be approximately 100 nm, containing typical EV markers such as Hsp70 and Tsg101. Deep sequencing analysis indicated the presence of 811 known and 15 novel miRNAs with an increasing number of differential miRNAs from the lung stage (27 miRNAs) to the liver stages (58 and 96 miRNAs at 14 and 21 dpi, respectively) in the plasma EVs of infected mice compared to EVs isolated from the uninfected control. In total, 324 plasma EV miRNAs were shown to be co-detected among different stages of infection and the validation of selected miRNAs showed trends of abundance similar to deep sequencing analysis. For example, miR-1a-3p and miR-122-5p showed higher abundance, whereas miR-150-3p and miR-126a showed lower abundance in the plasma EVs of infected mice at 3, 14, and 21 dpi as compared to those of uninfected mice. In addition, bioinformatic analysis combined with PCR validation of the miRNA targets, particularly those associated with the immune system and parasitic infectious disease, indicated a significant increase in the expression of Gbp7and Ccr5 in contrast to the decreased expression of Fermt3, Akt1, and IL-12a. Our results suggested that the abundance of miRNA cargo of the host plasma EVs was related to the stages of Schistosoma japonicum infection. Further studies on the roles of these miRNAs may reveal the regulatory mechanism of the host-parasite interaction. Moreover, the differentially abundant miRNA cargo in host EVs associated with S. japonicum infection may also provide valuable clues for identifying novel biomarkers for schistosomiasis diagnosis.

Taxa-specific activity loss and mortality patterns in freshwater trematode cercariae under subarctic conditions.

Cercarial activity and survival are crucial traits for the transmission of trematodes. Temperature is particularly important, as faster depletion of limited cercarial energy reserves occurs at high temperatures. Seasonal climate conditions in high latitude regions may be challenging to complete trematode life cycle during the 6-month ice-free period, but temperature effects on the activity and survival of freshwater cercariae have not been previously identified. After experimentally simulating natural subarctic conditions during warmer and colder months (13 and 6°C), a statistical approach identifying changes in the tendency of cercarial activity loss and mortality data was used to detect differences in three trematode genera, represented by four taxa (Diplostomum spp., Apatemon spp., small- and large-sized Plagiorchis spp.). A strong temperature-dependent response was identified in both activity loss and mortality in all taxa, with Diplostomum spp. cercariae showing the most gradual changes compared to other taxa. Furthermore, whilst activity loss and mortality dynamics could not be divided into ‘fish- vs invertebrate-infecting cercariae’ groups, the detected taxa-specific responses in relation to life-history traits indicate the swimming behaviour of cercariae and energy allocation among larvae individuals as the main drivers. Cercariae exploit the short transmission window that allows a stable continuance of trematodes’ life cycles in high-latitude freshwater ecosystems.

Phylogenetic position of Tremiorchis ranarum Mehra and Negi, 1926 (Trematoda: Plagiorchiidae) with remark on this monotypic genus.

Tremiorchis is a monotypic genus of digenetic trematode (Plagiorchiidae: Plagiorchiinae), infecting the frogs Rana tigrina (Hoplobatrachus tigerinus) and R. cyanophlyctis (Euphlyctis cyanophlyctis). Metacercaria use to infect Rana tigrina (Hoplobatrachus tigerinus) and R. cyanophlyctis (Euphlyctis cyanophlyctis) as intermediate hosts, while the cercaria stage found from apple snail, Pila virens. Adults of T. ranarum harbor mature frogs of H. tigerinus and E. cyanophlyctis. Besides the frequent infection of Tremiorchis, no DNA sequence data are currently available for this monotypic genus. The present communication, deals with the sequence data for nuclear ribosomal genes, 18S, small internal transcribed spacers (ITS1-5.8S-ITS2) and 28S to molecularly characterize T. ranarum. Besides this, phylogenetic relationship among the members of the Plagiorchiida is also discussed in detail. An attempt has also been made to provide detailed molecular affinities of T. ranarum with other trematode genera.

Morphological and molecular characterization of Quinqueserialis (Digenea: Notocotylidae) species diversity in North America.

Estimates of trematode diversity are inaccurate due to unrecognized cryptic species and phenotypic plasticity within species. Integrative taxonomy (genetics, morphology and host use) increases the clarity of species delineation and improves knowledge of parasite biology. In this study, we used this approach to resolve taxonomic issues and test hypotheses of cryptic species in a genus of trematode, Quinqueserialis. Specimens from throughout North America were field collected from hosts and obtained from museums. We found three morphologically distinct groups and successfully sequenced specimens from two of these groups. DNA sequencing at the 28S and CO1 gene regions revealed that two of the three groups were genetically distinct. One genetic group included two morphological clusters demonstrating host-induced phenotypic plasticity within Quinqueserialis quinqueserialis. The other unique genetic group is a novel species, Quinqueserialis kinsellai n. sp., which is described herein. Our study illustrates the importance of integrating multiple sources of evidence when investigating trematode diversity to account for the influence of cryptic species or phenotypic plasticity. However, further sampling is needed to understand Quinqueserialis spp. diversity as some species have no genetic information associated with them.

Three new species of Creptotrema (Trematoda, Allocreadiidae) with an amended diagnosis of the genus and reassignment of Auriculostoma (Allocreadiidae), based on morphological and molecular evidence.

Diversity of Creptotrema was investigated using morphological and molecular tools, including data for Creptotrema creptotrema (type-species). Three new species, parasites of Brazilian fishes, are described: Creptotrema conconae n. sp. (type-host, Imparfinis mirini Haseman), Creptotrema schubarti n. sp. (type-host, Characidium schubarti Travassos) and Creptotrema megacetabularis n. sp. (type-host, Auchenipterus osteomystax (Miranda Ribeiro)). The diagnosis of the genus was amended to include new features. The new species differ from each other mainly in terms of body shape, relative sucker size, and testes position. DNA sequences were obtained from Creptotrema spp. from Brazil, including 28S, ITS and COI. Genetic divergences among the new species and C. creptotrema varied from 2.1 to 5.2% (21–49 bp) for 28S, and 6.6 to 16.4% (21–45 bp) for COI. Phylogenetic analysis (28S) placed the newly generated DNA sequences of Creptotrema in a clade (Creptotrema clade sensu stricto) composed of C. creptotrema, the new species described herein, and all species previously described as Auriculostoma, revealing that Auriculostoma is best interpreted as a synonym of Creptotrema based on the principle of priority of zoological nomenclature. Creptotrema funduli, the single sequence of the genus previously available, was not grouped within the Creptotrema clade sensu stricto, suggesting the need for reevaluation of the taxonomic status of this species. Our results showed that Creptotrema represents a monophyletic genus of trematodes widely distributed across the Americas, which currently comprises 19 valid species of parasites of teleosts and anurans.

Host-induced phenotypic plasticity in Saccocoelioides lamothei Aguirre-Macedo and Violante-González, 2008 (Digenea: Haploporidae) a parasite of freshwater, brackish and marine fishes from Middle America.

Saccocoelioides is a genus of trematodes associated with fishes from the Americas. In the current research, morphologically distinct specimens of Saccocoelioides spp. were collected from six countries in Middle America. Specimens were sequenced using three molecular markers, the domains D1-D3 of the large subunit (LSU) from the nuclear rDNA, the cytochrome c oxidase subunit 1 (cox1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1) from mitochondrial DNA. A total of 74 new sequences were compared and aligned with other sequences available in GenBank. Maximum likelihood and Bayesian inference analyses were inferred from the LSU and cox1 datasets, revealing unequivocally that all the specimens correspond to S. lamothei. A haplotype network was built with 119 sequences of the nad1 gene. The network detected 57 distinct haplotypes divided into three haplogroups. To explore morphological differences among samples of S. lamothei, 17 morphological features were measured from 53 specimens from three fish families: Eleotridae, Mugilidae and Gobiidae. Principal component analysis yielded three main polygons that corresponded with each family analysed, suggesting host-induced phenotypic plasticity. The current evidence suggests that S. lamothei infects at least five fish families along the Pacific coasts of Mexico, Guatemala, El Salvador, Honduras, Nicaragua and Costa Rica.

An opisthorchiid concept of the genusLiliatrema(Trematoda: Plagiorchiida: Opisthorchioidea): an unexpected systematic position

AbstractLiliatrema is a small genus of trematodes consisting of two species. Its systematic position has long been debated, partly because of the confusing reports about the structure of male terminal genitalia. Here we test the phylogenetic position of the genus Liliatrema using data on complete 18S rRNA and partial 28S rRNA gene sequences obtained for Liliatrema skrjabini. We also provide a detailed description of terminal genitalia in adult specimens of L. sobolevi and metacercariae of both Liliatrema species. The results of the 28S rDNA-based phylogenetic analysis indicate that Liliatrema falls within a well-supported clade, which also includes Apophallus and traditional opisthorchiids. This clade, in turn, is nested within a well-supported clade, containing Euryhelmis, Cryptocotyle and Scaphanocephalus. In the 18S+8S rDNA analysis, Liliatrema appears as a sister-taxon to the Cryptocotyle + Euryhelmis group. The Liliatrema + (Cryptocotyle + Euryhelmis) clade is a well-supported sister-group to the traditional opisthorchiids. The morphology of the terminal genitalia of the liliatrematids also corresponds to that of the opisthorchioids. Thus, the results of our morphological and phylogenetic analyses favour an unexpected conclusion that the genus Liliatrema belongs to the Opisthorchioidea. We propose that the genera Liliatrema, Apophallus, Euryhelmis, Cryptocotyle and Scaphanocephalus belong, respectively, within the subfamilies Liliatrematinae, Apophallinae, Euryhelminthinae and Cryptocotylinae of the family Opisthorchiidae.