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The Cyathocotylidae Mühling, 1896 is a small but broadly distributed family of digeneans parasitic in a wide range of vertebrate definitive hosts, from fish to mammals. Despite existing taxonomic questions, only a few studies have generated DNA sequence data from cyathocotylids, and only a single publication included sequence data from a cyathocotylid parasitic in snakes. Four genera are known to infect snakes: GogateaLutz, 1935, Szidatia Dubois, 1938, MesostephanoidesDubois, 1951, and SerpentostephanusSudarikov, 1961. Members of these genera were known from only Asia and Africa. In the present study, we describe 2 new species of Gogatea from snakes in Australia and 1 from Vietnam. The new species from Vietnam described herein is the first member of the genus that lacks a ventral sucker. We used partial sequences of the nuclear large ribosomal subunit (28S) and mitochondrial cytochrome c oxidase subunit I (COI) genes to explore phylogenetic relationships among cyathocotylids and species differentiation. In addition, this is the first report of a cyathocotylid from snakes in Australia, thus posing interesting questions regarding the dispersal and historical biogeography of these parasites. Cyathocotylid genera from snakes have a long, convoluted taxonomic history. The genera Gogatea, Mesostephanoides, and Szidatia were distinguished from each other based on very small morphological differences. Therefore, the validity of Szidatia and Mesostephanoides was often questioned in the literature. Based on the detailed morphological analysis of our freshly collected high-quality specimens and comparison with published information, we synonymize Mesostephanoides and Szidatia with Gogatea., (© American Society of Parasitologists 2024.)

Haliotrema Pteroisi: Paperna, 1972 (Monogenoidea: Dactylogyridae) was found parasitizing the gill lamellae of devil firefish, Pterois miles (Bennet) (Perciformes: Scorpaenidae), in the Red Sea off Safaga (26°44'N, 33°56'E), Egypt. The parasite species was described based on morphological features of available specimens and transferred to PlatycephalotremaKritsky and Nitta, 2019 (Dactylogyridae) as Platycephalotrema pteroisi (Paperna, 1972) n. comb. The occurrence of Pl. pteroisi off Safaga, Egypt, represented a range extension for the helminth of about 160 km to the southwest of the southern end of the Gulf of Aqaba. The transfer of the species to Platycephalotrema based on an evaluation of morphological features was supported by an analysis of molecular sequences of the 28S rDNA gene of Pl. pteroisi and 49 other dactylogyrid species. Maximum-likelihood, Bayesian inference, and maximum parsimony analyses of this dactylogyrid sequence data revealed H. pteroisi to nest with significant support within the clade of Platycephalotrema spp. During the literature review of dactylogyrid species infecting scorpionfishes, it was determined that Ancyrocephalus sp. of Dyer et al. from luna lion fish Pterois lunulata Temminck and Schlegel collected off Okinawa-jima, Japan represented an undescribed species of Platycephalotrema., (© American Society of Parasitologists 2024.)

Environmental DNA (eDNA) surveys promise to be a sensitive and powerful tool for the detection of trematodes. This can contribute to the limited studies on trematode ecology, specifically in aquatic ecosystems. Here, we developed species-specific primer and probe sets for Moliniella anceps, Opisthioglyphe ranae, and Plagiorchis multiglandularis cercariae and applied a novel eDNA qPCR assay to detect larval trematodes quantitatively. We evaluated the effectiveness of the assays using filtered lake water samples collected from different sites of Lake Fadikha and Kargat River Estuary in Lake Chany, Russia, showing high species specificity and sensitivity in all 3 assays. Further, all 3 assays had high efficiencies ranging from 94.9 to 105.8%. Moliniella anceps, O. ranae, and P. multiglandularis were detected in the environmental water samples through real-time PCR. Thus, we anticipate that our approach will be beneficial for biomonitoring, measuring, and managing ecological systems., (© American Society of Parasitologists 2024.)

Some parasites manipulate their host's phenotype to enhance predation rates by the next host in the parasite's life cycle. Our understanding of this parasite-increased trophic transmission is often stymied by study-design challenges. A recurring difficulty has been obtaining uninfected hosts with a coevolutionary history with the parasites, and conducting experimental infections that mimic natural processes. In 1996, Lafferty and Morris provided what has become a classic example of parasite-increased trophic transmission; they reported a positive association between the intensity of a brain-infecting trematode (Euhaplorchis californiensis) in naturally infected California killifish (Fundulus parvipinnis) and the frequency of conspicuous behaviors, which was thought to explain the documented 10-30× increase in predation by the final host birds. Here, we address the primary gap in that study by using experimental infections to assess the causality of E. californiensis infection for increased conspicuous behaviors in F. parvipinnis. We hatched and reared uninfected F. parvipinnis from a population co-occurring with E. californiensis, and infected them 1-2 times/week over half their life span with E. californiensis and a small cyathocotylid trematode (SMCY) that targets the host's muscle tissue. At 3 time points throughout the hosts' lives, we quantified several conspicuous behaviors: contorting, darting, scratching, surfacing, and vertical positioning relative to the water's surface. Euhaplorchis californiensis and SMCY infection caused 1.8- and 2.5-fold overall increases in conspicuous behaviors, respectively. Each parasite was also associated with increases in specific conspicuous behaviors, particularly 1.9- and 1.4-fold more darting. These experimental findings help solidify E. californiensis-F. parvipinnis as a classic example of behavioral manipulation. Yet our findings for E. californiensis infection-induced behavioral change were less consistent and strong than those previously documented. We discuss potential explanations for this discrepancy, particularly the idea that behavioral manipulation may be most apparent when fish are actively attacked by predators. Our findings concerning the other studied trematode species, SMCY, highlight that trophically transmitted parasites infecting various host tissues are known to be associated with conspicuous behaviors, reinforcing calls for research examining how communities of trophically transmitted parasites influence host behavior., (© American Society of Parasitologists 2023.)

The identity of a larval bucephalid infecting the eastern oyster, Crassostrea virginica (Gmelin, 1791), from a tidal river in Virginia is investigated using ribosomal deoxyribonucleic acid (rDNA). The internal transcribed spacer (ITS1, 5.8S, ITS2) region and a portion of the 28S rDNA were isolated from genomic DNA from sporocysts containing cercariae and compared with sequences from GenBank and from our previous collections of possibly related bucephalids. The studied larval bucephalid was 100% identical at the ITS1, 5.8S, and partial 28S rDNA sequences with Prosorhynchoides paralichthydis (Corkum, 1961) Curran and Overstreet, 2009; however, it differed from P. paralichthydis by 6 bases plus 3 deletions in the ITS2 region. This level of variation at the ITS2 region has been demonstrated among some Indo-Pacific species of Prosorhynchoides Dollfus, 1929, suggesting that the larval bucephalid represents an unidentified or innominate species of Prosorhynchoides that is closely related to P. paralichthydis., (© American Society of Parasitologists 2023.)

A new digenean species belonging in EncyclobrephusSinha, 1949 is described, and the generic diagnosis is amended to accommodate variation in several features of the new species. Worms were collected from the intestines of 2 specimens of the Mekong snail-eating turtle, Malayemys subtrijuga (Schlegel and Müller, 1845). Permanent whole-mounted worms were studied using light microscopy, and ribosomal DNA (rDNA) sequences were generated from 3 worms. We investigated the phylogenetic relationship of the new species among some digenea using separate Bayesian inference analyses, 1 based on the 28S rDNA gene and rooted using a species from the Monorchioidea Odhner, 1911, and a second based on the internal transcribed spacer 1 region rooted by a species in the Microphalloidea Ward, 1901. Prior to the analyses, Encyclobrephus was classified in the Encyclometridae Mehra, 1931. Previous studies using rDNA from the type species for the family, Encyclometra colubrimurorum (Rudolphi, 1819) Baylis and Cannon, 1924, have demonstrated that En. colubrimurorum is closely related to species of Polylekithum Arnold, 1934 in the Gorgoderoidea Looss, 1901. Nevertheless, phylograms from both analyses indicated that the new species of Encyclobrephus belongs in the Plagiorchioidea Lühe, 1901, related to species in the families Cephalogonimidae Looss, 1899, Plagiorchiidae Lühe, 1901, Reniferidae Pratt, 1902, and Telorchiidae Looss, 1899. The present results suggest that Encyclobrephus is not closely related to En. colubrimurorum. Familial classification of Encyclobrephus is contingent on molecular data availability for the type species but it should be removed from the Encyclometridae and classified as incertae sedis within the Plagiorchioidea. Encyclometridae belongs in the Gorgoderoidea, not the Plagiorchioidea., (© American Society of Parasitologists 2023.)

We herein resurrect and emend PlesiocreadiumWinfield, 1929 (Digenea: Macroderoididae) and provide a supplemental description of its type species, Plesiocreadium typicumWinfield, 1929, based on adult specimens collected from the intestine of bowfins, Amia calva Linnaeus, 1766 (Amiiformes: Amiidae), captured in the L'Anguille River (Mississippi River Basin, Arkansas), Big Lake (Pascagoula River Basin, Mississippi), Chittenango Creek (Oneida Lake, New York), and Reelfoot Lake (Tennessee River Basin, Tennessee). Plesiocreadium spp. (Pl. typicum and Plesiocreadium flavum [Van Cleave and Mueller, 1932] n. comb.) differ from other macroderoidids by having a dorsoventrally flat forebody, ceca that extend posteriad beyond the testes and that do not form a cyclocoel, testes that are greater than one-half of maximum body width, a cirrus sac that is dorsal to the ventral sucker and arches dextrad or sinistrad, a uterine seminal receptacle, asymmetrical vitelline fields that remain separated anteriorly and posteriorly and that extend anteriad to the level of the ventral sucker, and an I-shaped excretory vesicle. Bayesian phylogenetic analyses (ITS2 and 28S) recovered monophyletic Plesiocreadium sensu stricto (as defined herein) sister to Macroderoides trilobatusTaylor, 1978 and that clade sister to the remaining macroderoidids, with sequences ascribed to species of Macroderoides Pearse, 1924 recovered as paraphyletic. We regard Macroderoides parvus (Hunter, 1932) Van Cleave and Mueller, 1934, M. trilobatus, and RauschiellaBabero, 1951 as incertae sedis. Arkansas, New York, and Tennessee comprise new locality records for Pl. typicum., (© American Society of Parasitologists 2023.)

A survey was conducted in the lowland areas of Binh Dinh and Phu Yen provinces in south central Vietnam to study the diversity of digenean cercariae within freshwater snails. These regions are known as highly endemic areas for the transmission of liver flukes, Fasciola spp. and Opisthorchis viverrini. In total, 13 snail species were found, and 3 of these were dominant, comprising 51.9% of all collected snails, i.e., Bithynia funiculata, Melanoides tuberculata, and Radix auriculata. Cercariae were found in 5 snail species, and among these, Indoplanorbis exustus had the highest overall prevalence of infection (38.5%). No cercarial infections were found in viviparid and ampullariid snails. Morphological identification resulted in 10 cercariae types that were illustrated and measured. Among them, xiphidiocercariae and echinostomata were the most common, causing 70.55% of all cercariae infections. Bithynia species had the most diverse digenean fauna. Molecular data revealed the presence of cercariae of 13 digenean species, including O. viverrini. The 18S rDNA sequences of echinostomata and xiphidiocercariae type 3, which were shed from different snail hosts, were similar to 2 and 3 digenean species, respectively. In conclusion, our findings highlight the diversity of digenean fauna in Binh Dinh and Phu Yen provinces., (© American Society of Parasitologists 2022.)

Most trematodes and some cestodes have obligate life history features that include an asexual developmental stage that can produce genetically-identical individuals (clonemates) followed by an adult stage with sexual reproduction. These life history features can influence the evolutionary mechanism of inbreeding in parasites, especially among self-compatible hermaphroditic endoparasites whose mating opportunities are restricted to within hosts. As clonemate mating in hermaphroditic species produces a genetic inbreeding signature identical to that of self-mating, it is important to understand how clonemates are transmitted through their life stages. A handful of prior studies compared clone richness (number of clones) across life cycle stages to infer transmission processes and to characterize clone abundance distributions (CADs) among hosts. Here we illustrate the use of the proportion of clonemate dyads (PC) within hosts to describe the CADs. PC has several advantages as an ecological metric in that it is unbiased by sample size, takes into account relative parasite burdens, and has a direct transmission interpretation, i.e., the probability of cotransmitting clonemates. Moreover, PC is also an evolutionary metric as it can be used to estimate a potential clonemate mating rate. We demonstrate the use of PC in comparing CADs within and across 2 trematode developmental stages in the lancet fluke Dicrocoelium dendriticum. Also, we show how genetic estimates of apparent selfing (true selfing plus clonemate mating) at larval and juvenile stages can be compared to PC estimated at the adult stage to assess the contribution of clonemate mating to apparent selfing. The eco-evolutionary links presented are generalizable to assess sibling cotransmission as well. Thus, the framework presented herein will facilitate future field-based studies on the transmission and mating systems of parasitic flatworms., (© American Society of Parasitologists 2022.)

We revise monotypic LeuceruthrusMarshall and Gilbert, 1905 (Azygiidae Lühe, 1909) by emending its generic diagnosis, redescribing its type species (Leuceruthrus micropteriMarshall and Gilbert, 1905), reassigning 2 species (Leuceruthrus stephanocauda [Faust, 1921] n. comb., Leuceruthrus ocalana [Smith, 1935] n. comb.), describing 2 new congeners (Leuceruthrus ksepkai n. sp. and Leuceruthrus blaisei n. sp.), and providing a phylogenetic analysis based on the internal transcribed spacer 2 (ITS2). Leuceruthrus is unique by having oblique preovarian testes, a vitellarium that does not extend anteriad into the forebody, and a uterus that is intercecal and between the ovary and ventral sucker. We describe the cercaria of Leuceruthrus cf. stephanocauda from cercariae shed from Elimia cf. carinifera and Elimia cf. modesta from Big Canoe Creek, Alabama. Leuceruthrus ksepkai n. sp. is described from cercariae shed from rasp elimia, Elimia floridensis (Reeve, 1860) from Holmes Creek, Florida, and Elimia sp. 1 from the Chocktawhatchee River, Florida. It differs from its congeners by the combination of having broadly rounded furcae with slight marginal pigmentation in live cercariae, no spines on the tail stem, distinct anterior and posterior ridges that flank the tail stem portion accommodating the withdrawn distome, minute protuberances occupying the lateral margin of the tail stem for its entire length, and protuberances that encircle the anterior third of the posterior tail stem (immediately posterior to the tail stem portion containing the withdrawn distome). Leuceruthrus blaisei n. sp. infects Elimia sp. and is the only known congener having proportionally small furcae (>tail stem maximum width) with a nipple-like distal projection and numerous minute projections on the tail stem surface. Our phylogenetic analysis included all of our new sequences plus all publicly available ITS2 sequences for Leuceruthrus spp. and Proteromera spp., and supported the monophyly of Leuceruthrus. It recovered L. ksepkai and L. cf. ksepkai in a polytomy, and Leuceruthrus blaisei as monophyletic (identical sequences), with L. micropteri and L. cf. stephanocauda recovered as sister taxa. The present study comprises the first systematic treatment of Leuceruthrus in over a century, the first description of a new species of Leuceruthrus in 117 yr, and the first taxonomic characterization of a species of Leuceruthrus from the Mobile River Basin. It also increases the number of accepted congeners from 1 to 5 and provides new host records (E. cf. carinifera and E. floridensis) for Leuceruthrus spp., (© American Society of Parasitologists 2022.)

Two cases of human philophthalmosis have been reported in Japan. Gravid flukes removed from the eyes of the patients were broken, but their morphological characteristics suggest that an unknown species of the genus Philophthalmus is involved as a pathogen for humans. The mitochondrial DNA barcode of the human eye fluke enabled us to discover its larval stage from the Japanese mud snail, Batillaria attramentaria. The discovered cercaria had previously been temporarily described as "Philophthalmid sp. I." In this study, we examined the infection status of B. attramentaria with Philophthalmid sp. I found on a muddy seashore of the Seto Inland Sea, Japan, and the resulting metacercariae were experimentally administered to Japanese quails to develop them into the gravid adult stage. The complete specimens of the adult and larval stages allowed us to describe a new species. Based on morphological and molecular analyses, Philophthalmus hechingeri n. sp. is proposed for the human-infecting eye fluke in Japan. The natural definitive hosts of the new species are unknown. However, the habitat of B. attramentaria suggests that shorebirds (seagulls, sandpipers, and plovers) might be the possible candidates., (© American Society of Parasitologists 2022.)

Parasitism is one of the most common consumer strategies and contributes a large portion to biological diversity. Trematodes in the family Diplostomidae are common in freshwater ecosystems worldwide, often residing in the eyes or brain of fish and then infecting fish-eating birds as adults. As a result, some species have broad geographic distributions due to the bird host's motility. In contrast to the cosmopolitan nature of diplostomids, only a single species, Tylodelphys darbyi, has been identified in New Zealand to date, and only from the South Island. Tylodelphys darbyi has a 3-host life cycle consisting of an unidentified snail, a freshwater fish (Gobiomorphus cotidianus), and the Australasian crested grebe (Podiceps cristatus australis). To date, T. darbyi has been found in 2 locations, Lake Hayes, in the eyes of G. cotidianus, and Lake Wanaka, adults recovered from grebes. Considering the near ubiquity of the fish host in New Zealand, it is likely the bird, listed as nationally vulnerable, is the limiting factor in the range of T. darbyi. Up to 10 G. cotidianus were sampled from 10 mountain lakes known to have populations of grebe in the Otago and Canterbury regions of New Zealand's South Island. The eyes of all fish were examined and any metacercariae present were set aside for genetic analysis. In addition to expanding the known range of T. darbyi to at least 4 water bodies across the South Island, 2 new taxa of diplostomid were identified. A lens-infecting metacercariae clustered with Diplostomum spathaceum, while the metacercariae from the humor clustered with Diplostomum baeri., (© American Society of Parasitologists 2021.)

This study investigated the identity of 2 lepocreadiid digenean species belonging in the genus Opechona Looss, 1907 that infect littoral fishes of the northern Gulf of Mexico. Opechona chloroscombriNahhas and Cable, 1964, a species previously known only from the Caribbean Sea and the Atlantic Ocean off Brazil, is reported herein from the Atlantic bumper, Chloroscombrus chrysurus (L.), in the northern Gulf of Mexico. A second species infects the gulf butterfish, Peprilus burti Fowler, and the American harvestfish, Peprilus paru (L.), and it is described as a new species that occurs in coastal waters of the north-central and northeastern Gulf of Mexico. Metacercariae infecting the mesoglea of pelagic jellyfishes (Bougainvillia carolinensis [McCady], Chrysaora quinquecirrha [Desor], and Stomolophus meleagris Agassiz) and pelagic comb jellies (Mnemiopsis leidyi Agassiz and Beroe ovata Bruguière) were collected that resemble the new species but require further study to identify. Newly generated sequence fragments (28S rDNA) from both species of Opechona plus 2 other lepocreadiids collected during the study were aligned with publicly available sequences from 18 other lepocreadiids, 6 species of Aephnidiogenidae Yamaguti, 1934, and 2 species of Gorgocephalidae Manter, 1966. The alignment was subjected to Bayesian inference analysis rooted using a gorgocephalid. The resulting tree estimated the positions of both Opechona spp. as being unresolved within a group of taxa that included all available species of Opechona plus available species from the morphologically similar genera ProdistomumLinton, 1910, Preptetos Pritchard, 1960, and Clavogalea Bray, 1985. Although relatively similar in morphology, the 2 studied species of Opechona were surprisingly not closely related. Opechona cablei (Stunkard, 1980) Bray and Gibson, 1990 is herein considered to be a junior synonym of Opechona pyriformis (Linton, 1900) Bray and Gibson, 1990., (© American Society of Parasitologists 2021.)

Neopsilotrema is a small genus of psilostomid digeneans parasitic in the intestine of birds in the Palearctic and Nearctic. At present, the genus includes 4 species: Neopsilotrema lisitsynae from the Palearctic and Neopsilotrema affine, Neopsilotrema lakotae, and Neopsilotrema marilae from the Nearctic. Herein, we describe a new species, Neopsilotrema itascae n. sp., from lesser scaup Aythya affinis collected in Minnesota. The species can be distinguished from congeners on the basis of the ventral sucker:oral sucker width ratio, body width:length ratio, and cirrus sac size, along with other characters. We generated new 28S ribosomal deoxyribonucleic acid (DNA) and NADH dehydrogenase (ND1) mitochondrial DNA sequence data of a variety of psilostomids from the Palearctic and Nearctic along with sequences of the ribosomal internal transcribed spacer (ITS) region (ITS1 + 5.8S + ITS2) from 3 Neopsilotrema species. The molecular phylogenetic affinities of a variety of psilostomid taxa were studied using 28S sequence data. The 28S sequences of psilostomids demonstrated 1-7.9% intergeneric divergence, whereas the sequences of ND1 had 17.7-34.1% intergeneric divergence. The interspecific divergence among members of Neopsilotrema was somewhat lower (0.2-0.5% in 28S; 0.3-0.4% in ITS; 12-15.7% in ND1). Our comparison of DNA sequences along with morphologic study suggests Holarctic distribution of N. lisitsynae., (© American Society of Parasitologists 2021.)

Herein we describe a new species and propose a new genus, Posthovitellinum psiloterminae n. gen., n. sp. (Lissorchiidae: Asymphylodorinae), based on specimens that infect the intestine of Cyclocheilos enoplos (Bleeker, 1849) (Cypriniformes: Cyprinidae), a migratory riverine carp from the Mekong River (Dong Thap province, Vietnam). The new species is assigned to Lissorchiidae by having a combination of features: spinous tegument, subterminal oral sucker, pre-equatorial ventral sucker, median and pretesticular ovary, submarginal genital pore at level of the ventral sucker, follicular vitellarium distributing in 2 lateral fields, and lacking eyespot pigment in the adult. It cannot be assigned to any existing asymphylodorine genus because it has the combination of a well-developed cirrus-sac, an unarmed ejaculatory duct and metraterm, a follicular vitellarium distributing in 2 lateral fields located between the posterior margin of the ventral sucker and the mid-level of the testis, and a sinistral, submarginal genital pore. The new species has an elongate, claviform cirrus-sac, a single, large, elongate-oval testis at the posterior extremity of the body, operculate eggs, and an I-shaped excretory bladder with secondary branches at the level of the testis and extending anteriad to the level of the pharynx. Bayesian inference analysis of the partial large subunit ribosomal DNA gene (28S rDNA) recovered the new species sister to Asaccotrema vietnamienseSokolov and Gordeev, 2019; these species differed by 118 nucleotides (12%; 983 bp fragment). This is the first lissorchiid reported from the Mekong River; only the second from southern Vietnam; and the fourth reported from a cyprinid fish in Vietnam. The aforementioned phylogenetic analysis included previously unpublished sequences representing lissorchiids infecting the intestine of North American suckers (Cypriniformes: Catostomidae): Lissorchis cf. nelsoni from spotted sucker; Minytrema melanops (Rafinesque, 1820) and Lissorchis cf. gullaris (immature) from smallmouth buffalo, Ictiobus bubalus (Rafinesque, 1818). Asymphylodora atherinopsidisAnnereaux, 1947, herein is treated as a species incertae sedis. The 28S tree topology suggests that Lissorchiinae may comprise more than 1 lineage, but additional species are needed to confidently assert this., (© American Society of Parasitologists 2021.)

As presently recognized, Herpetodiplostomum is a small genus of proterodiplostomid digeneans parasitic in crocodilians. Most members of Cheloniodiplostomum, a genus of proterodiplostomids that parasitize chelonians, were originally placed within Herpetodiplostomum. The 2 genera were distinguished based on the presence/absence of papillae on the holdfast organ and anterior extent of vitellarium. Our study of Herpetodiplostomum and Cheloniodiplostomum revealed a lack of consistent morphological differences between the genera. Therefore, we consider Cheloniodiplostomum to be a junior synonym of Herpetodiplostomum. Recent molecular phylogenetic studies included a number of proterodiplostomid taxa; however, DNA sequence data are not available for any Herpetodiplostomum or Cheloniodiplostomum species. Herein, we describe a new Herpetodiplostomum species from Geoffroy's side-necked turtle Phrynops geoffroanus from Mato Grosso State, Brazil. The new species can be distinguished from other Herpetodiplostomum species based on the distribution of vitellarium, level of development of holdfast papillae, and ratio of prosoma:opisthosoma width, among other characters. We provide the first molecular phylogeny of the Proterodiplostomidae that includes a Herpetodiplostomum species. The limited geographic distribution of Herpetodiplostomum spp. within the geographic ranges of caimans suggests a secondary host switching event from crocodilians to chelonian definitive hosts in the evolution of Herpetodiplostomum., (© American Society of Parasitologists 2021.)

The hawksbill turtle Eretmochelys imbricata is a critically endangered species with a worldwide distribution. Limited information is available about the naturally occurring intestinal parasites of this species and what impact these parasites may have on the health of the hawksbill turtle. Diaschistorchis pandus was identified postmortem in 5 hawksbill turtles from Grenada, West Indies, using morphologic characterization. Sanger sequencing was performed for conserved ribosomal regions (5.8S, ITS2, 28S) and the mitochondrial cytochrome c oxidase subunit 1 gene (COI). Phylogenetic analysis of the 28S rRNA gene sequence data shows D. pandus clustering with other trematodes in the family Pronocephalidae, corroborating morphological classification. No genetic sequences have been previously reported for this trematode species, which has limited the collection of objective epidemiological data about this parasite of marine turtles., (© American Society of Parasitologists 2021.)

Cardicola Short, 1953 is the most speciose aporocotylid genus (35 species) and includes marine and estuarine species of fish blood flukes that infect "higher ray-finned fishes" (Euteleostei). Several clades within Cardicola are recovered in phylogenetic analyses of the large subunit ribosomal DNA (28S), but morphological synapomorphies for those nucleotide-based clades remain elusive. The type species, Cardicola cardiocola (Manter, 1947) Short, 1953, has not been recollected in 73 yr and the original description was incomplete; making a genus revision challenging because of the ambiguous systematic position of its type species. Herein, we redescribe C. cardiocola by using the holotype (USNM 1337732) and new specimens collected from the type host, jolthead porgy, Calamus bajonado (Sparidae), from nearby the type locality. It differs from its congeners by the combination of having a body that is 5 times longer than wide, an anterior sucker with concentric rows of spines, 2-6 tegumental body spines per row, an esophageal gland that is 22-43% of the esophageal length, a testis that is 3-5 times longer than wide and that fills the intercecal space, a vitelline duct connecting to the anterior aspect of the oötype, an ascending uterus that lacks any coil, a descending uterus yielding a single coil, an obvious cirrus sac separated by constriction from the seminal vesicle, a tegumental protrusion surrounding the terminal end of cirrus sac, and a male genital pore that is posterior to the remainder of the genitalia. We also describe a new congener infecting the heart of yellowedge grouper, Hyporthodus flavolimbatus (Serranidae), from the Gulf of Mexico. It differs from its congeners by the combination of having an anterior sucker that does not extend beyond the anterior body margin, 2-5 tegumental body spines per row, posterior ceca that are 9 times length of the anterior ceca and that lack any coil, a testis that is 3 times longer than wide and that does not fill the intercecal space, an ovary that is >60% of the body width, a vitelline duct that connects to the anterior aspect of the oötype, a uterus that is >10% of the body width and that extends posterior to all genitalia, and a rounded posterior body margin. It is the first species of Cardicola to be described from a grouper (Serranidae). The 28S and internal transcribed spacer 2 phylogenetic analyses recovered the new species as a distinct lineage within the clade of Cardicola spp., (© American Society of Parasitologists 2021.)

The genus LangeroniaCaballero and Bravo-Hollis, 1949, currently contains 6 species of amphibian trematodes distributed in North and Middle America. The type species of the genus, Langeronia macrocirraCaballero and Bravo-Hollis, 1949, occurs in Mexico and is relatively commonly found as a parasite of leopard frogs. However, information regarding its life cycle is lacking. In this paper, we study the life cycle of L. macrocirra in Laguna Escondida, Los Tuxtlas, Veracruz. Definitive hosts (Rana spp.) as well as potential intermediate hosts (gastropods, bivalves, crustaceans, tadpoles, hemipterans, and odonate naiads) were sampled in the locality and studied to search for the presence of adults and larval stages of the trematode. Specimens were morphologically characterized, and some individuals were sequenced for 1 ribosomal gene (28S rRNA) and 1 mitochondrial gene (COI). DNA sequences of the adults obtained from leopard frogs were matched with those of the larval forms in their intermediate hosts (metacercariae, cercariae, and sporocysts) to demonstrate conspecificity. Further, we conducted a detailed study of the tegument of the body surface with scanning electron microscopy to characterize each of the developmental stages of the life cycle of L. macrocirra., (© American Society of Parasitologists 2020.)

A survey of the parasite fauna of freshwater fishes from the Wet Tropics Bioregion in Queensland, Australia, revealed the presence of a new species of Stemmatostoma Cribb, 1986 (Digenea: Cryptogonimidae). Stemmatostoma cribbi n. sp. is described from the intestine and pyloric caeca of 2 species of grunter (Terapontidae), Hephaestus fuliginosus (Macleay) and Hephaestus tulliensis (De Vis), and the Jungle perch (Kuhliidae), Kuhlia rupestris (Lacepède), collected from the Barron and Mulgrave-Russell River drainage divisions in tropical north Queensland, Australia. Stemmatostoma cribbi is primarily distinguished morphologically from the type and only other species in the genus, Stemmatostoma pearsoni Cribb, 1986, in having consistently fewer oral spines (14 in S. cribbi vs. 16 in S. pearsoni). Alignment of novel molecular data for S. cribbi and S. pearsoni revealed that they differ genetically by 26 nucleotides (2.1%) over the 1,258 bp partial large subunit (LSU) region, 1 nucleotide (0.8%) over the 121 bp partial 5.8S region, and 23 nucleotides (7.2%) over the entire 318 bp ITS2 rDNA region. Bayesian inference and maximum likelihood phylogenetic analyses of the partial LSU region for the species of Stemmatostoma sequenced here were used to explore the relationships of these species to other cryptogonimid species reported from freshwater ecosystems., (© American Society of Parasitologists 2020.)

In an ongoing investigation on the helminths of amphibians in southeastern Mexico, specimens of 2 undescribed species of Haematoloechus were collected from Rana brownorum . Haematoloechus ceciliae n. sp. is morphologically most similar to Haematoloechus meridionalis , but differs in the shape of the oral sucker, in the nature of the acetabulum, and in the distribution of the glandular cells in the pharyngeal region; Haematoloechus celestunensis n. sp. closely resembles Haematoloechus floedae , but differs in the form and size of the testes and measurements of acetabulum. COI and 28S DNA sequences of both new species show high divergence compared to other species of the genus. In the phylogenetic trees, H. ceciliae appears most closely related to Haematoloechus danbrooksi and H. celestunensis to Haematoloechus veracruzanus .

Codonocephalus is a monotypic genus of diplostomid digeneans and is the only genus in the sub-family Codonocephalinae. The type-species Codonocephalus urniger has an unusual progenetic metacercaria that uses frogs as intermediate hosts and can use snakes as paratenic hosts. Adult C. urniger parasitize ardeid wading birds in the Palearctic. Despite the broad distribution of Codonocephalus , no DNA sequence data are currently available for the genus. In this study, we generated sequence data for nuclear ribosomal and mitochondrial DNA from progenetic metacercaria of the type-species C. urniger from marsh frog, Pelophylax ridibundus , collected in Ukraine. We used partial sequences of the nuclear ribosomal 28S gene to examine for the first time the phylogenetic position of Codonocephalus among the Diplostomoidea.

Uvulifer Yamaguti, 1934, is a genus of diplostomoidean digeneans that parasitizes kingfishers worldwide. Species have a Neascus -type metacercaria that encysts in or on fish intermediate hosts, often causing black spot disease. Only 3 prior studies published DNA sequence data for Uvulifer species with only 1 including a single named species ( Uvulifer spinatus López-Jiménez, Pérez-Ponce de León, & García-Varela, 2018). Herein we describe 2 new species of Uvulifer from the green-and-rufous kingfisher, Chloroceryle inda (Linnaeus), collected in Peru ( Uvulifer batesi n. sp. and Uvulifer pequenae n. sp.). Both new species are readily differentiated from their New World congeners by a combination of morphological characters including distribution of vitelline follicles and prosoma:opisthosoma length ratios. In addition, we used newly generated nuclear 28S rRNA and mitochondrial COI gene sequence data to differentiate among species and examine phylogenetic affinities of Uvulifer . This includes the 2 new species and Uvulifer ambloplitis (Hughes, 1927), as well as Uvulifer elongatus Dubois, 1988 , Uvulifer prosocotyle (Lutz, 1928), and Uvulifer weberi Dubois, 1985 , none of which have been part of prior molecular phylogenetic studies. Our data on Uvulifer revealed 0.1-2.2% interspecific divergence in 28S sequences and 9.3-15.3% in COI sequences. Our 28S phylogeny revealed at least 6 well-supported clades within the genus. In contrast, the branch topology in the COI phylogenetic tree was overall less supported, indicating that although COI sequences are a great tool for species differentiation, they should be used with caution for phylogenetic inference at higher taxonomic levels. Our 28S phylogeny did not reveal any clear patterns of host association between Uvulifer and particular species of kingfishers; however, it identified 2 well-supported clades uniting Uvulifer species from distant geographical locations and more than 1 biogeographic realm, indicating at least 2 independent dispersal events in the evolutionary history of the New World Uvulifer . Our results clearly demonstrate that the diversity of Uvulifer in the New World has been underestimated.

Herein we describe a new species of turtle blood fluke (TBF) and propose a new genus to accommodate it, Pitiutrema revelae n. gen., n. sp. This blood fluke infects the heart of six-tubercled Amazon River turtles ( Podocnemis sextuberculata [Cornalia, 1849] [Pleurodira: Podocnemididae]) in the headwaters of the Amazon River near Iquitos, Peru. It resembles the other 2 described species of South American freshwater TBFs ( Atamatam amazoniensis Bullard and Roberts, 2019, and Paratamatam iquitosiensis Bullard and Roberts, 2019) by having a dorsoventrally flattened and ovoid body, an oral sucker with anteroventral spines, 2 inter-cecal testes arranged in a column, inter-gonadal terminal genitalia, an inter-cecal and post-ovarian Laurer's canal pore, and a Y-shaped excretory bladder. It differs from all other nominal TBFs by having the combination of an aspinose body that lacks mammillae, a tapered (not broadly rounded) posterior body end, a ventral sucker, slightly M-shaped or inverse U-shaped ceca, a deeply-lobed (dendritic) ovary, a transverse uterus, and a dispersed vitellarium. The new genus is further unique among TBF genera by having an anterior to posterior sequence of ventral sucker, anterior testis, ovary, cirrus sac (lateral to posterior half of ovary), and posterior testis. The phylogenetic results and placement of the new taxon (1) were both predicted by our morphological diagnosis and comparisons with related taxa, (2) further indicated monophyly of the nominal South American freshwater TBFs, (3) reaffirmed the marine derived lineage identity of the nominal South American freshwater TBFs, and (4) highlighted that the single cercarial sequence (TBF sp. W-810) from an ampullariid in Brazil does not share a recent common ancestor with any of the nominal South American freshwater TBFs. The new species is the eighth TBF reported from a side-necked turtle (Pleurodira), the first TBF from a member of Podocnemididae, and the third freshwater TBF from South America.

Philophthalmid eyeflukes are cosmopolitan parasites of birds and occasionally of mammals, including humans. A gravid adult of Philophthalmus sp. was found from the bulbar conjunctiva of a 64-yr-old woman in Japan, who was diagnosed with acute conjunctivitis. The parasite was morphologically most similar to Philophthalmus hegeneri , but distinctive in lacking an esophagus and in having clearly lobed testes. The DNA sequence analysis of genes for nuclear 28S ribosomal RNA and mitochondrial cytochrome c oxidase subunit 1 supported the identification at generic level. The morphological and molecular analyses strongly suggest that the eyefluke from a human in Japan should be treated as an undescribed species of Philophthalmus . The occurrence of human philophthalmosis is very rare. As far as we know, a total of 11 human cases have been reported worldwide to date.

Two new genera and species of freshwater turtle blood flukes (TBFs) are described herein based on specimens infecting the nephritic and mesenteric blood vessels of "matamatas" (a side-necked turtle, Chelus fimbriata [Schneider, 1783] [Pleurodira: Chelidae]) from the Amazon River Basin, Peru. These taxa comprise the first-named species and the first-proposed genera of freshwater TBFs from the continent of South America. A new comparison of all TBF genera produced 6 morphologically diagnosed groups that are discussed in light of previous TBF classification schemes and a novel phylogenetic hypothesis based on the nuclear large subunit ribosomal DNA (28S). Considering external and internal anatomical features, species of the new genera ( Atamatam Bullard and Roberts n. gen., Paratamatam Bullard and Roberts n. gen.) are most similar to each other and are together most similar to those of several marine TBF genera. The 28S phylogenetic analysis supported the monophyly of all 6 morphologically diagnosed groups of genera. Most notably, the freshwater TBFs of South America comprise a derived group nested within the clade that includes the paraphyletic marine TBFs. Not surprisingly in light of morphology, another marine TBF lineage ( Neospirorchis Price, 1934) clustered with the freshwater TBFs of Baracktrema Roberts, Platt, and Bullard, 2016 and Unicaecum Stunkard, 1925. Our results, including an ancestral state reconstruction, indicated that (1) freshwater TBFs have colonized marine turtles twice independently and that (2) the South American freshwater TBFs comprise a marine-derived lineage. This is the first evidence that TBFs have twice independently transitioned from a marine to freshwater definitive host. Marine incursion is considered as a possible mechanism affecting the natural history of marine-derived freshwater TBFs in South America. A dichotomous key to accepted TBF genera is provided.

Parasites can affect animal populations and communities in aquatic ecosystems. However, greater understanding is needed for the distributions and drivers of parasite infection levels in many areas. This study focuses on parasite prevalence (percent infected hosts) of an important class of parasites, trematodes, in 2 species of snail first intermediate hosts ( Planorbella trivolvis and Physa sp.) in the Illinois River watershed, which has been impacted by human development. We hypothesized that trematode prevalence depends on local (e.g., water chemistry) and landscape (e.g., proximity to the Illinois River and land cover) factors. To test our hypotheses, we collected at least 20 individuals of 1 or both species of snails from 28 ponds within the watershed, and we made water-quality measurements and recorded habitat characteristics at each site. We then screened the snails for infections in the laboratory and identified the trematode cercariae that emerged based on morphological and molecular techniques. We found 5 cercariae morphotypes, including important parasites of wildlife, such as Echinostoma sp. and Ribeiroia ondatrae . Our results indicate that proximity to the Illinois River and open water or wetlands was positively associated with trematode prevalence in both snail species, whereas water chemistry (higher pH, lower calcium concentration, and lower specific conductance) was associated with increased prevalence, but only in Physa . Our findings offer increased understanding of potential environmental drivers underlying trematode distributions, with implications for wildlife health.

Learedius learedi Price, 1934 , is a blood fluke found in sea turtles, and the adult fluke parasitizes the cardiovascular system of the host. In this study we surveyed 46 green sea turtles, Chelonia mydas , on the Ogasawara Islands, Japan, and blood flukes were detected in the heart and blood vessels of 26 turtles. The flukes were identified as L. learedi based on a detailed morphological description. In addition, molecular identification and characterization of the parasite were performed. The nucleotide sequences of nuclear internal transcribed spacer 2 ( ITS2 ) regions were almost identical to those of L. learedi reported previously, but not to those of Hapalotrema spp., which is the closest related genus. The nucleotide sequences of the 28S ribosomal DNA region formed a single clade with those of the reference L. learedi in the phylogenetic tree, but not with those of Hapalotrema spp. Therefore, the nucleotide sequences of ITS2 and 28S are robust markers for distinguishing L. learedi from other species. The nucleotide sequences of the mitochondrial cytochrome c oxidase subunit 1 ( COI ) region were analyzed to evaluate the genetic variations in L. learedi . The COI haplotypes revealed the extremely high genetic diversity of the species as well as the host turtles on the Ogasawara Islands. The haplotype frequency in the mitochondrial DNA of the green sea turtles on the Ogasawara Islands is known to be significantly different from those in other Pacific rookeries. Although the number of analyzed flukes is small in this study, no haplotype was close to that in other areas; on the basis of the data, we hypothesized that L. learedi differentiated along with the host turtles on the Ogasawara Islands.

Adults of the genus Austrodiplostomum are parasites in cormorants of the New World, whereas metacercariae are parasites from eye globe and brain of freshwater and brackish water fishes. In this study, specimens of Austrodiplostomum mordax from South America (type-species) were analyzed together with other specimens of Austrodiplostomum spp. collected from several locations across Middle America and North America. Partial DNA sequences of the mitochondrial gene cytochrome c oxidase subunit I ( COI), the internal transcribed spacers ( ITS1, ITS2, and 5.8S gene), and the D2-D3, domains of the large subunit ( LSU) of nuclear ribosomal DNA, were generated for both developmental stages and compared with available sequences of Austrodiplostomum spp. Phylogenetic analyses inferred with each molecular marker using maximum likelihood and Bayesian inference revealed the existence of 4 lineages representing 2 described species, A. mordax and Austrodiplostomum compactum (syn. Austrodiplostomum ostrowskiae) and 2 undescribed species of Austrodiplostomum recognized in previous studies. The COI haplotype network inferred with 172 sequences detected 28 haplotypes divided into 4 clusters, separating each other by 33 and 40 substitutions and with a genetic divergence ranging from 9 to 12%. The largest group included specimens identified as A. compactum plus those identified as A. ostrowskiae, supporting the synonymy of both species. As a result, we conclude that A. compactum is widely distributed across the Americas, in locations of the United States, Mexico, El Salvador, Honduras, Costa Rica, Venezuela, Peru, and Brazil. The other 2 undescribed species of the genus Austrodiplostomum were previously recorded in the United States and now are reported in Mexico. These 2 species cannot be described because adult forms have not been found in their definitive hosts. Finally, the species A. mordax has been found only in some lakes from Argentina, and it was validated in this study through molecular analyses.

The freshwater fish digenean Pseudosellacotyla lutzi ( Freitas, 1941 ) Yamaguti, 1954 has had an unsettled taxonomic history, and has at various times been classified as a member of Nanophyetidae, Heterophyidae, Microphallidae, Faustulidae, and Cryptogonimidae. Nine individual specimens of the trahira, Hoplias malabaricus (Bloch, 1794), were sampled in the Paraná River basin, Paraná State, Brazil; 22 specimens of P. lutzi were collected. One specimen of P. lutzi was used to obtain a sequence of the domains D1-D3 of the 28S rRNA gene, and to perform a phylogenetic analysis to assess their position and classification within Plagiorchiida. The resulting tree unequivocally shows that the species, along with acanthostomines, belong to the Cryptogonimidae, corroborating recent findings based on the morphology of the cercariae, and in the characteristics of the life cycle. In addition, the study of the ultrastructure of the tegumental spines through scanning electron microscopy allowed us to characterize them as pectinate spines possessing 3 to 8 digitiform projections at their distal end and extending from the anterior to the posterior extremity of the body. This study also provides the first molecular data for a cryptogonimid from South America.

The phylogenetic position of Clinostomum heluans Braun, 1899 within the genus Clinostomum Leidy, 1856 is reported in this study based on sequences of the barcoding region of the mitochondrial cytochrome c oxidase subunit 1 gene ( COX1). Additionally, molecular data are used to link the adult and the metacercariae of the species. The metacercariae of C. heluans were found encysted infecting the cichlid fish Australoheros sp. in Minas Gerais, Brazil, whereas the adults were obtained from the mouth cavity of the Great White Egret, Ardea alba, in Campeche, Mexico. The COX1 sequences obtained for the Mexican clinostomes and the Brazilian metacercaria were almost identical (0.2% molecular divergence), indicating conspecificity. Similar molecular divergence (0.2-0.4%) was found between sequences of C. heluans reported here and Clinostomum sp. 6 previously obtained from a metacercaria recovered from the cichlid Cichlasoma boliviense in Santa Cruz, Bolivia. Both maximum likelihood and Bayesian inference analyses unequivocally showed the conspecificity between C. heluans and Clinostomum sp. 6, which form a monophyletic clade with high nodal support and very low genetic divergence. Moreover, tree topology reveals that C. heluans occupies a basal position with respect to New World species of Clinostomum, although a denser taxon sampling of species within the genus is further required. The metacercaria of C. heluans seems to be specific to cichlid fish because both samples from South America were recovered from species of this fish family, although not closely related.

Two cryptic species of haploporid digeneans belonging in Saccocoelioides are described from Costa Rica: one from a poeciliid fish, Poecilia gillii, and the other from a characid fish, Astyanax aeneus. Traditional morphological features are examined and found to be largely inefficient for differentiating among the new species and 20 New World congeners. Comparison of ribosomal DNA sequences among the 2 new species, 2 North American, 4 Middle American, and 3 South American species of Saccocoelioides, including the type-species Saccocoelioides nanii from Argentina and Saccocoelioides sogandaresi from Texas, is effective in differentiating among these species. A Bayesian inference analysis is conducted using a concatenated alignment of the same 2 ribosomal gene regions from 35 species belonging in the Haploporoidea and rooted by the atractotrematid Isorchis anomalus. The analysis provides stronger support for a close relationship between species in Saccocoelioides and Intromugil; thus, Intromugil is transferred from the Waretrematinae to the Chalcinotrematinae. Additionally, interrelationships among 11 species of Saccocoelioides are resolved and reveal a distinct genetic rift suggesting the presence of 2 distinct lineages within the genus, 1 containing the type-species and the other containing a group of species that more closely resemble other genera in the subfamily. Taxonomy of Saccocoelioides is discussed, and observations made during the present study justify taking several taxonomic actions: new combinations are proposed for Saccocoelioides tilapiae n. comb., formerly in the now-defunct Culuwiya, and Saccocoelioides ruedasueltensis n. comb., formerly in Chalcinotrema; Saccocoelioides guaporense nomen novem is proposed for Lecithobotrioides elongatus; Saccocoelioides papernai is considered a junior subjective synonym of Saccocoelioides overstreeti; Saccocoelioides godoyi is considered a junior subjective synonym of Saccocoelioides szidati; Saccocoelioides magnorchis and Saccocoelioides saccodontis are considered species inquirendae; and Saccocoelioides adelae is considered a nomen nudum. Four new hosts are reported for Saccocoelioides cichlidorum: Amphilophus lyonsi, Amatitlania nigrofasciatus, Amatitlania septemfasciatus, and Hypsophrys nicaraguensis. Molecular data call into question some existing species identifications in Saccocoelioides and reveal that molecular tools combined with traditional taxonomy are required for accurately identifying species in the genus. Twenty-two species of Saccocoelioides are formally accepted, but it is noted that as new molecular data become available, some of these species may be transferred to other genera in the subfamily.

Plagioporus ictaluri n. sp. and Plagioporus carolini n. sp. are described from the intestines of the Ouachita Madtom, Noturus lachneri, and the Banded Sculpin, Cottus carolinae, respectively, from adjacent drainages in Arkansas. The new species are morphologically most similar to one another and in turn similar to Plagioporus sinitsini, Plagioporus chiliticorum, Plagioporus serratus, and Plagioporus hypentelii, but they can be distinguished from these congeners in possession of an excretory vesicle that extends anteriorly to the level of the anterior testis as opposed to 1 reaching only the posterior testis ( P. hypentelii) or 1 confined to the posttesticular space ( P. sinitsini, P. serratus, and P. chiliticorum), a feature that necessitates altering the generic diagnosis for the genus. Plagioporus ictaluri n. sp. is distinguished from Plagioporus carolini n. sp. in having tandem vs. oblique testes, a submedian to median ovary as opposed to 1 that is dextral, a ventral sucker occupying 53-71% of the body width (BW) vs. 80-92% of the BW, an oral sucker occupying 36-47% of the BW as opposed to 49-58% of the BW, and a pharynx occupying 21-26% of the BW compared to 28-36% of the BW. A Bayesian inference analysis of partial 28S rDNA sequences of the 2 new species and those of 34 opecoelids obtained from GenBank found that P. ictaluri and P. carolini formed a highly supported clade that was sister to P. chiliticorum and in turn to P. sinitsini. These 4 species are notably the only Nearctic plagioporids included in the analysis without a uterus extending to the posterior end that lack a confluent vitelline field in the posttesticular space. This study includes the first species of Plagioporus to be described from an ictalurid host and the first species in the genus to be described from a cottid east of the Rocky Mountains.

Neospirorchis (Digenea: "Spirorchiidae") are blood flukes of sea turtles. Trematodes tentatively identified as Neospirorchis sp. infect various sites within sea turtles inhabiting waters of the southeastern United States, but efforts to obtain specimens adequate for morphologic study has proven difficult. Two genetic targets, the internal transcribed spacer region of the ribosomal RNA gene and the partial mitochondrial cytochrome c oxidase subunit I gene, were used to investigate potential diversity among parasite specimens collected from stranded sea turtles. Sequence data were obtained from 215 trematode and egg specimens collected from 92 individual free-ranging cheloniid sea turtles comprising 4 host species. Molecular analysis yielded more than 20 different genotypes. We were able to assign 1 genotype to 1 of the 2 recognized species, Neospirorchis pricei Manter and Larson, 1950 . In many examples, genotypes exhibited host and site specificity. Our findings indicate considerable diversity of parasites resembling Neospirorchis with evidence of a number of uncharacterized blood flukes that require additional study.

The diversity and taxonomy of diplostomids infecting freshwater fishes of the Indian region are predominantly poorly known. Yet compared to other trematodes, identification and discrimination of diplostomid metacercaria are difficult using morphology and site of infection. The present study provides the first molecular study of a Tylodelphys sp. from India. Specimens of Tylodelphys were collected from the vitreous humor of the eyes and from the brains of siluriform fish, Mystus tengara (Siluriformes: Bagridae) from Hastinapur, Meerut (U.P.), India. Taxonomic studies were performed on the basis of morphological, morphometrical, and molecular analysis. The dissimilarities in morphological and sites of infection between 2 forms of Tylodelphys, designated as Tylodelphys sp. 1 and Tylodelphys sp. 2, suggested that the forms were different species but, after performing molecular analysis, we conclude that despite morphological differences both morphotypes are conspecific.

This study investigates the systematic position of some species of the genus Saccocoelioides Szidat, 1954, and the species Culuwiya cichlidorum Aguirre-Macedo and Scholz, 2005 , from North and Middle America using new morphological data and partial sequences of 2 nuclear genes, the large subunit and the internal transcribed spacer 2 from ribosomal DNA. In total 74 specimens representing 4 species of Saccocoelioides (41 of Saccocoelioides sogandaresi, 3 of Saccocoelioides chauhani, 6 of Saccocoelioides lamothei, and 12 of Saccocoelioides olmecae) plus 12 specimens of Culuwiya cichlidorum were analyzed. The phylogenetic analyses inferred with maximum likelihood method and Bayesian inference showed that the 4 species of Saccocoelioides formed a clade divided into 4 subclades representing each species. In addition, 7 specimens sequenced from the type host and locality of Culuwiya cichlidorum were nested in 2 clades: the first clade with specimens of C. cichlidorum from Costa Rica, and the second clade with specimens of S. sogandaresi from Mexico, Honduras, Nicaragua, and Costa Rica. Our analyses suggest that C. cichlidorum should be reallocated into Saccocoelioides, as was originally designated Saccocoelioides cichlidorum n. comb., is restricted to Nicaragua and Costa Rica and associated with cichlid fishes. In contrast, the specimens identified previously as Culuwiya cichlidorum from the coast of Gulf of Mexico by Aguirre-Macedo and Scholz belong to S. sogandaresi, which has a wide distribution that extends from Galveston Bay (U.S.A.) in Gulf of Mexico to Atlantic and Pacific slopes associated with poeciliid and cichlid fishes. New morphological data derived from scanning electron microscopy photographs of the body surface show a characteristic pattern of spination that is useful to distinguish between S. cichlidorum n. comb. and S. sogandaresi.

Metacercarial stages of digeneans were collected from decapod crustaceans inhabiting intertidal rocky zones in central Chile. The digeneans were identified through a molecular analysis based on the V4 region of the 18S ribosomal RNA gene. We analyzed 356 crustaceans belonging to 10 species, 115 intertidal fish belonging to 6 species, and 4 specimens of 1 coastal bird species. In total, 74.1% of crustaceans were parasitized with metacercariae. We found 1 species of Opecoelidae. This species showed low genetic divergence (0% and 0.1%) with adult digeneans found in intertidal fish and with the species Helicometrina labrisomi infesting a subtidal fish from northern Chile (Labrisomus philippii). Additionally, we found 2 species of Microphallidae, 1 closely related to Maritrema (1.3% genetic distance) and the other related to Microphallus (5% genetic distance). Therefore, our findings showed that the decapod crustaceans are relevant hosts in food webs from the southeastern Pacific coast. Furthermore, we found 5 species of crustaceans as second intermediate hosts for H. labrisomi and 2 species as secondary intermediate hosts for 2 Microphallidae, which contribute to elucidate parts of their life cycles through molecular markers and extended the host distribution of H. labrisomi in the southeastern Pacific coast.

:  Baracktrema obamai n. gen., n. sp. infects the lung of geoemydid turtles (black marsh turtle, Siebenrockiella crassicollis [type host] and southeast Asian box turtle, Cuora amboinensis ) in the Malaysian states of Perak, Perlis, and Selangor. Baracktrema and Unicaecum Stunkard, 1925 are the only accepted turtle blood fluke genera having the combination of a single cecum, single testis, oviducal seminal receptacle, and uterine pouch. Baracktrema differs from Unicaecum by having a thread-like body approximately 30-50× longer than wide and post-cecal terminal genitalia. Unicaecum has a body approximately 8-12× longer than wide and terminal genitalia that are anterior to the distal end of the cecum. The new genus further differs from all other accepted turtle blood fluke genera by having a cecum that is highly convoluted for its entire length, a spindle-shaped ovary between the cirrus sac and testis, a uterine pouch that loops around the primary vitelline collecting duct, a Laurer's canal, and a dorsal common genital pore. Phylogenetic analysis of the D1-D3 domains of the nuclear large subunit ribosomal DNA (28S) revealed, with high nodal support and as predicted by morphology, that Baracktrema and Unicaecum share a recent common ancestor and form a clade sister to the freshwater turtle blood flukes of Spirorchis, paraphyletic Spirhapalum, and Vasotrema and that, collectively, these flukes were sister to all other tetrapod blood flukes (Hapalorhynchus + Griphobilharzia plus the marine turtle blood flukes and schistosomes). Pending a forthcoming emended morphological diagnosis of the family, the clade including Spirorchis spp., paraphyletic Spirhapalum, Vasotrema, Baracktrema, and Unicaecum is a likely placeholder for "Spirorchiidae Stunkard, 1921 " (type genus Spirorchis MacCallum, 1918 ; type species Spirorchis innominatus Ward, 1921 ). The present study comprises the 17th blood fluke known to infect geoemydid turtles and the first proposal of a new genus of turtle blood fluke in 21 yr.

:   Infidum similis Travassos, 1916 (Dicrocoeliidae: Leipertrematinae) was found in the gall bladder of Leptophis diplotropis Günther, 1872 from El Podrido, Acapulco, Guerrero, Mexico. A phylogenetic analysis based on partial sequences of the 28S ribosomal RNA using maximum likelihood (ML) and Bayesian inference (BI) analyses was carried out to assess its phylogenetic position within suborder Xiphidiata, alongside members of the superfamilies Gorgoderoidea and Plagiorchoidea. The phylogenetic trees showed that the genus is most-closely related to the Plagiorchoidea rather than to the Gorgoderoidea, in keeping with previous taxonomic designations. Phylogenies obtained from ML and BI analysis of the 28S rDNA gene revealed a well supported clade in which Choledocystus hepaticus (Lutz, 1928) Sullivan, 1977 is sister to I. similis. On the other hand, a tree obtained using a partial sequence of the cytochrome c oxidase subunit 1 (cox1) mtDNA gene (ML and BI analysis), with species supposed to be closely related to I. similis according to 28S, does not support this relatedness. Based on the independence of Infidum from the subfamily Leipertrematinae Yamaguti, 1958 , our results clearly demonstrated that the genus corresponds to a different family and with species closely related to C. hepaticus within Plagiorchoidea. New data are presented about the tegumental surface of I. similis by scanning electron microscopy as well as the estimation of its haploid genome size using Feulgen Image Analysis Densitometry of sperm nuclei as part of the characterization of this species. This is the first genome size estimated for a member of Plagiorchiida, and these data will provide a new source of knowledge on helminth diversity and evolutionary studies. This constitutes the first host record, and new geographical distribution, for this species in Mexico.

An undescribed, cryptic species of Didymocystis, as determined from sequences of 2 ribosomal genes and superficially similar to Didymocystis scomberomori ( MacCallum and MacCallum, 1916 ), infected the skin of the Spanish mackerel, Scomberomorus maculatus , in the north-central Gulf of Mexico (GOM). An analysis of 558 fish from 2011 to 2013 from Louisiana, Mississippi, Alabama, and the Florida panhandle showed the prevalence of the trematode varied both spatially and temporally but not with sex of the fish host. Month, year, and geographic location were identified by a negative binomial generalized linear model as indicators of the abundance and intensity of infection. Prevalence, abundance, and intensity of infection were greatest in spring and fall months off the Florida panhandle. Furthermore, the abundance and intensity of infection correlated negatively with fork length, weight, and gonad weight of mature fish but positively with longitude. Therefore, smaller adult fish tended to be more infected than larger adults, and prevalence and intensity increased from west to east (Louisiana to Florida). Spatial and temporal trends seemed to result from physical factors (e.g., water temperature, salinity, bottom type), but they also coincided with the annual migration of S. maculatus as fish moved northward along the GOM coastline from the southern tip of Florida in the spring months and returned in the fall, being present in the north-central GOM from late spring through fall. This pattern suggests the possibility that acquisition of infections occurred from a molluscan host in waters off the Florida panhandle.

Metacercariae of species of Euclinostomum have been found encysted in kidney, liver, and muscles of several fish species, while adults occur in the mouth, pharynx, larynx, trachea, and upper esophagus of fish-eating birds. The aim of this work was to gather molecular and morphological data from the type species, Euclinostomum heterostomum, as a starting point for needed revisions of the genus. Metacercariae were collected from cichlids in Lake Kinneret, Israel, and all were identified as E. heterostomum based on morphology. This identification was further confirmed by principal components analyses that revealed no partitions in morphometric resemblance, indicating that the material represents a single species, and that the specimens were morphometrically similar to other descriptions of E. heterostomum. In phylogenetic analysis of concatenated partial internal transcribed spacer 1 and 2, 5.8S rDNA, and partial cytochrome c oxidase I barcode sequences, the isolates of Euclinostomum obtained in this study form a single, monophyletic group separate from recently published data from Euclinostomum sp. from Thailand and from Clinostomum species, which are also monophyletic. The morphological and molecular data reported in the present work can provide a useful point of reference for future studies.

Ocular diplostomiasis is caused by trematode species in the family Diplostomidae, specifically those in the genera Austrodiplostomum, Diplostomum, and Tylodelphys. Diplostomid trematodes are globally distributed parasites of fish. Heavy infections of diplostomids that parasitize the eyes of fish can result in acute mortality while chronic infections are often characterized by impaired vision or blindness. In the southeastern United States, commercial catfish production is threatened by piscivorous birds and the many trematode species that parasitize them. The life cycles typically involve a piscivorous avian definitive host, a mollusk first intermediate host, and a fish second intermediate host. A survey of parasites infecting the snail host Biomphalaria havanensis (= B. obstructa ) in catfish production ponds was undertaken. Snails were collected from 2 separate ponds during the summer of 2014 and observed for the release of trematode cercariae. A total of 1,740 snails were collected. Three distinct longifurcate pharyngeate cercariae were observed and these cercariae were characterized morphologically and molecularly. Sequencing of ∼4,200 base pairs (bp) of the nuclear ribosomal genes and ∼450 bp of the mitochondrial cytochrome c oxidase gene revealed 3 genetically distinct species. One morphotype shared 99-100% sequence identity with metacercariae from the aqueous and vitreous humors of gizzard shad Dorosoma cepedianum and channel catfish Ictalurus punctatus as well as an adult trematode, Austrodiplostomum ostrowskiae, a parasite of the double-crested cormorant Nannopterum auritus. The remaining 2 cercariae morphotypes shared 99-100% sequence identity with an unidentified Tylodelphys sp. and Austrodiplostomum sp. metacercaria from the brain and eyes of several freshwater fish. Herein we molecularly link the cercaria, metacercaria, and adult stage of the life cycle of A. ostrowskiae, identifying the snail host for this parasite, in addition to providing notes on 2 cercariae representing 2 other diplostomids.

This study investigates the systematics of Paracreptotrema Choudhury, Pérez-Ponce de León, Brooks and Daverdin, 2006 using morphological data (stained whole mounts and scanning electron microscopy) and partial sequences of the 28S ribosomal rRNA gene, obtained from freshly collected material. In total, 484 specimens representing 4 species, i.e., Paracreptotrema blancoi (157), Paracreptotrema profundulusi (12), Paracreptotrema rosenthali (8), and Paracreptotrema blancoi sensu Salgado-Maldonado et al. (2011) (307) were collected. Existing museum depositions were also studied. The 28S rRNA gene sequences of these Paracreptotrema spp. were aligned, along with sequences from 22 other allocreadiids and 4 other non-allocreadiid xiphidiatan species. Bayesian inference and maximum likelihood analyses indicated a paraphyletic Paracreptotrema split into 3 clades: 1 comprising P. blancoi and P. rosenthali that was sister to a clade formed by 3 other species of allocreadiids (species of Wallinia, Creptotrematina, and Auriculostoma) typically found in characid fishes, a second clade formed solely by Paracreptotrema heterandriae as the sister taxon of the aforementioned species, and a third by P. profundulusi and specimens erroneously identified as P. blancoi. Two new taxa were erected to reflect these results: Paracreptotrematoides for Paracreptotrema heterandriae, and Pseudoparacreptotrema for Paracreptotrema profundulusi and P. macroacetabulata (the species erroneously identified as P. blancoi from profundulids across Middle America). Closer consideration of the morphology corroborates these findings. The revised systematics also indicated that Paracreptotrema spp. are found in poeciliids, whereas Pseudoparacreptotrema spp. parasitize profundulids. The study demonstrates the value of an integrative taxonomy approach to address the apparently complicated systematics of the allocreadiids.

A complete morphological description, supported by molecular data, of Clinostomum detruncatum metacercariae is reported in this study. The metacercariae were found infecting Synbranchus marmoratus from the Paraná River, municipality of Guaíra, Paraná State, Brazil. The taenioidean uterus is the main morphological characteristic of this species. Both neighbor-joining (NJ) and maximum likelihood (ML) trees showed 2 clear separate clusters grouping together the species from the Palearctic region (Clinostomum complanatum, Clinostomum cutaneum, Clinostomum phalacrocoracis, Clinostomum philippinense) and those from the Nearctic/Neotropical regions (Clinostomum marginatum, Clinostomum tataxumui, C. detruncatum, Clinostomum sp. 7). The latter clade is divided into 2 subclades grouping species from North America and Mexico (C. marginatum and C. tataxumui), and those from Brazil (C. detruncatum and Clinostomum sp. 7).

Two species of digeneans belonging in Homalometron are described from Nearctic freshwater fundulid fishes: Homalometron robisoni n. sp. is described from the Blackstripe Topminnow, Fundulus notatus, from Oklahoma and Homalometron frocioneae n. sp. is described from the Banded Killifish, Fundulus diaphanus, from New York. Homalometron robisoni n. sp. differs from all congeners by having vitelline follicles that extend into the forebody, a feature that necessitates altering the generic diagnosis for the genus. Homalometron frocioneae n. sp. may be distinguished from North and Middle American congeners by the position of the intestinal bifurcation (relatively more posterior in the forebody than in other species) and tegumental spine coverage on the body (spines are absent from the posterior body extremity and on most of the dorsal surface). Comparison of ribosomal DNA (ITS 1 and 2 regions, 5.8S gene, and partial fragment of 28S gene) from the 2 new species and some congeners from the Western Hemisphere provided evidence for the validity of the 2 new species and affirmed a close relationship between H. robisoni n. sp. and Homalometron pallidum. Comparison of ribosomal DNA from newly collected Homalometron spp. and larval stages of an apocreadiid from brackish water hydrobiid snails (cercariae in rediae in Littoridinops palustris and metacercariae in L. palustris and Amnicola limosa ) from a tidal river in Mississippi revealed that larval stages represented Homalometron cupuloris. A phylogeny based on Bayesian inference analysis using partial 28S rDNA gene fragments from 14 species of Homalometron (all from the Western Hemisphere) and 1 megaperine and rooted by a second megaperine was conducted and produced a strongly supported phylogram that estimates the interrelationships among species. The estimated phylogeny suggests that ecological factors such as salinity and food web interactions between species of Homalometron, intermediate hosts, and fishes drive coevolutionary forces influencing speciation within Homalometron.

The opecoelid Helicometrina nimia Linton, 1910 has been reported from numerous marine fishes along the Pacific and Atlantic coasts of the Americas. Along the Chilean coast, H. nimia is found in fishes belonging to at least 9 families. This surprisingly low host specificity of H. nimia raises question about the correct identification of specimens assigned to this species. Here we evaluate whether H. nimia specimens isolated from sympatric fish species in northern Chile but with different diets and found in different habitats (water column and demersal) are the same species. Our results demonstrate that specimens from the shallow benthic fish Labrisomus philippii (Steindachner) do not correspond to H. nimia but instead belong to a new species of Helicometrina. This species is described and distinguished from H. nimia using morphological descriptions and 2 molecular markers (the cytochrome c oxidase subunit 1 gene and the V4 region of the SSU rRNA gene). The new species Helicometrina labrisomi (Digenea: Opecoelidae), is found in the intestine of L. philippii (Steindachner, 1866) (Pisces: Labrisomidae), a shallow benthic fish that inhabits the northern coast of Chile. We also studied the related Helicometrina nimia Linton, 1910 from the benthopelagic fishes Paralabrax humeralis (Valenciennes, 1828) and Acanthistius pictus (Tschudi, 1846) (Serranidae). The new species differs from H. nimia by a combination of characters that include ovary shape, number of uterine loops, and position of the genital pore. Our results indicate that morphological characteristics, such as body size, extent of the vitellarium, shape of the testes, and cirrus sac size and extent, traditionally used in the taxonomy of Helicometrina are highly variable. In contrast, meristic and morphological characteristics, such as a lobed ovary, the number of uterine loops, dimensions of the pharynx, and the opening of the genital pore, are highly constant.

In a study of 106 sunfish, Mola mola (L.), from Mediterranean waters, 2,731 worms, belonging to 2 congeneric species of accacoeliids, Accacladocoelium macrocotyle (Diesing, 1858) Robinson, 1934 and Accacladocoelium nigroflavum (Rudolphi, 1819) Robinson, 1934 , were collected from the digestive system. It is often difficult to differentiate between these 2 species as they are sympatric and very similar; in fact, according to previous descriptions, the extent of the vitellarium is the only interspecific difference, described as extending posteriorly to the ovary in A. macrocotyle and as anterior to the anterior testis in A. nigroflavum. However, this diagnostic trait is not always valid; moreover, it is often indistinguishable because it is masked by the uterine eggs. Here, the morphology of new specimens of both species has been studied in detail and combined with molecular analysis. This study shows that the extension of the vitellarium is very similar in both species, but the degree of ramification differs. Furthermore, although the morphological differences are very slight, both species seemed genetically different: intraspecific differences ranged between 0.8 and 1.5% in internal transcribed spacer (ITS)-2 rRNA gene (ITS2) and between 0.5 and 1.6% in cytochrome c oxidase subunit 1 (COI) and interspecific differences ranged between 2 and 3.2% in ITS2 and between 9.6 and 10.6% in COI. In addition, we observed previously undescribed morphological differences, which help to differentiate these 2 species: the oral sucker is relatively smaller in A. nigroflavum than in A. macrocotyle and the ovary is usually relatively longer in A. nigroflavum than in A. macrocotyle.

Despite the large number of reports of species of Clinostomum from vertebrate hosts in South America, studies evaluating the molluscan transmitters of these parasites are scarce. In the present study, clinostomatoid cercariae shed from 0.02% (4/17,485) specimens of Biomphalaria spp., collected at the Pampulha reservoir, Belo Horizonte, Minas Gerais, Brazil, were used for experimental infection of Poecilia reticulata . Samples of cercariae from molluscs and metacercariae experimentally obtained from fish were subjected to morphological and molecular analyses and compared with species of Clinostomum reported in the Americas. The cercariae and metacercariae, here identified as Clinostomum sp., present general morphology similar to that reported for Clinostomum marginatum , however, from molecular point of view, differ significantly from North American C. marginatum and other species of Clinostomum reported in South America. These results suggest that the diversity of Clinostomum found in Brazil may be underestimated. Additional studies aimed at molecular characterization of South American species of Clinostomum, including the finding of specimens with sequences similar to that reported for C. marginatum in North America are required.

Classification of most monogeneans is primarily based on size, shape, and arrangement of haptoral sclerites. These structures are often obscured or misinterpreted when studied using light microscopy, leading to confusion regarding defining characters. Scanning electron microscopy (SEM) has predominantly been used to study haptoral sclerites in smaller monogeneans, focusing on hooks and anchors. In the Diplozoidae, SEM has not been used to study haptoral sclerites. Using new and modified techniques, the sclerites of diplozoids collected in South Africa were successfully studied using SEM. The digestion buffer from a DNA extraction kit was used to digest the surrounding tissue, and Poly-L-lysine-coated and concavity slides were employed to limit the movement and loss of sclerites, with the latter being more user-friendly. In addition to the success of visualizing the sclerites using SEM, the digested tissue from as little as half of the haptor provided viable genetic material for molecular characterization. From the results presented here, the study of the sclerites of larger monogeneans using SEM, including those bearing clamps, is a viable possibility for future research. Also, this method may be beneficial for the study of other, non-haptoral sclerites, such as cirri in other families of monogeneans. During this study, Labeo capensis was noted as a valid host of Paradiplozoon vaalense in a region of the Vaal River where the type host, Labeo umbratus, appears to be absent.

Mesodesma donacium is a dominant species on sandy beaches along the Chilean coast. However, the only previous parasite records for this species were obtained for the northern Chilean coast (20° S-33° S), which dealt with cestodes, polychaetes, and copepods. In this study, the symbiotic fauna of M. donacium in its southern distributional range is reported, and the geographical variations in the occurrence of this fauna are evaluated. A total of 565 individuals of M. donacium were captured by local fishermen from 5 localities: Mehuín (MEH) (39° 26' S), Carelmapu (CAR) (41° 44' S), Ancud (ANC) (41° 53' S), Cucao (CUC) (42° 35' S), and Quellón (QUE) (43° 24' S), covering a total distance of 450 km. To collect metazoan symbionts, the valves, mantle, gills, gonad, and digestive gland of each specimen of yellow clam were examined, and symbiont identifications were made via morphological and genetic analyses. The prevalence and mean intensity of infestation were calculated for each symbiotic species. Univariate and multivariate analyses were performed to evaluate the differences in symbiotic load between localities. Seven metazoan symbiotic species were recorded. The most abundant species were Paranthessius mesodesmatis, Monorchiidae gen. sp., and Paravortex sp. The copepod P. mesodesmatis and metacercaria Monorchiidae gen. sp. showed a high prevalence in all localities, but their intensity of infection varied among localities. The turbellarian Paravortex sp. was most frequently associated with ANC and CUC. The digenean Sanguinicolidae gen. sp. was recorded only at CAR, and the polychaete Spionidae gen. sp. was recorded only at MEH. In its southern distributional range, M. donacium was characterized by an absence of cestodes. This absence can be explained by the absence of the definitive host. The local environmental conditions in the southern range of the host could explain the differences in symbiotic composition among localities.