Redescription of Dracovermis occidentalis (Digenea: Liolopidae) infecting American alligator, Alligator mississippiensis from the Bon-Secour River (Mobile-Tensaw River Delta, Alabama, USA) and a revised phylogeny for Liolopidae.

We examined several American alligators, Alligator mississippiensis (Daudin, 1802) (Crocodilia: Alligatoridae) from Louisiana, Alabama, and South Carolina in August 2022. The intestine of one alligator from Alabama was infected by Dracovermis occidentalis Brooks and Overstreet, 1978 (Platyhelminthes: Digenea: Liolopidae Odhner, 1912), a seldom collected and incompletely described trematode that lacks a representative nucleotide sequence. Liolopidae comprises 5 genera and 15 species: Liolope spp. infect giant salamanders; Helicotrema spp. infect turtles and lizards; Harmotrema spp. infect snakes; Paraharmotrema spp. infect turtles; and Dracovermis spp. infect crocodilians. Based on our study of the newly collected specimens and the holotype of D. occidentalis, we redescribe D. occidentalis, correct errors in its original description, and provide an updated phylogeny for Liolopidae that, for the first time, includes Dracovermis Brooks and Overstreet, 1978. Our specimens were identified as D. occidentalis by having testes in the posterior 1/3 of the body, a pretesticular cirrus sac, a spined and eversible cirrus, a bipartite seminal vesicle, and a post-acetabular vitellarium. A phylogenetic analysis of the D1-D3 domains of the nuclear large subunit ribosomal DNA (28S) recovered Liolopidae as monophyletic; however, low taxon sampling in the group precludes hypothesis-testing about liolopid-vertebrate cophyly. This is the first collection for morphological study of the type species for Dracovermis since the genus was proposed 46 years ago, the first record of a liolopid from Alabama, and the first phylogenetic analysis that includes Dracovermis.

First report of Myxobolus neurofontinalis (Bivalvulida: Myxobolidae) infecting anadromous Brook Trout from Prince Edward Island, Canada.

OBJECTIVE: During routine histological examination of tissues from mortality events of anadromous Brook Trout Salvelinus fontinalis from Prince Edward Island (PEI), Canada, myxospores consistent with Myxobolus were observed infecting the central nervous system. The objective of this study was to identify the species of Myxobolus infecting the nervous system of anadromous Brook Trout from PEI, Canada. METHODS: Myxospore morphology, small subunit (SSU) ribosomal DNA (rDNA) sequence data, and histology were used to identify myxospores isolated from infected Brook Trout. RESULT: Myxospore measurements from the PEI samples matched those reported in the description of Myxobolus neurofontinalis from North Carolina. A 1057-bp fragment of the SSU rDNA from myxospores collected from Brook Trout in PEI was identical to an isolate of M. neurofontinalis (MN191598) collected previously from the type locality, New River basin, North Carolina. Histological sections confirmed infections were intercellular in the central nervous system. Minimal host response was observed, with only sparse mononuclear inflammatory infiltrates present at the periphery of and within dispersed myxospores, suggesting that infections are not pathogenic to Brook Trout. CONCLUSION: Myxospores were identified as M. neurofontinalis, which was previously described from the central nervous system of Brook Trout from the New River basin, North Carolina, USA. This constitutes the first time M. neurofontinalis has been documented outside of the New River basin in North Carolina.

A new species of Ellipsomyxa Køie, 2003 (Bivalvulida) infecting the gall bladder of Pangasius macronema Bleeker (Siluriformes: Pangasiidae) from the Mekong River Delta, Vietnam.

During a parasitological survey of freshwater fishes in the Mekong River Delta, Vietnam, disporic plasmodia containing myxospores morphologically consistent with Ellipsomyxa Køie, 2003 (Bivalvulida) were observed infecting the gall bladder of Pangasius macronema Bleeker (Siluriformes: Pangasiidae). Herein, we use morphology and small subunit ribosomal DNA (SSU rDNA, 18S) sequence data to describe Ellipsomyxa intravesica Ksepka & Bullard n. sp. and relate it to other myxozoans. The new species resembles Ellipsomyxa adlardi Whipps & Font, 2013, which infects the naked goby, Gobiosoma bosc (Lacepede) (Gobiiformes: Gobiidae) in Lake Pontchartrain, Louisiana, but differs from it by having a longer myxospore (mean = 13.3; range = 12.0-15.0 vs. 12.4; 11.3-14.4) and shorter polar capsules (3.7; 3.0-4.0 vs. 4.3; 3.9-4.9). The 18S phylogenetic analysis recovered the sequence of the new species sister to those ascribed to Ellipsomyxa ariusi Chandran, Zacharia, Sathianandan & Sanil, 2020 and Ellipsomyxa sp. (MK561979); both of which infect the gall bladder of the threadfin sea catfish, Arius arius (Hamilton) (Siluriformes: Ariidiae) from the southwest coast of India. Consistent with previous phylogenetic analyses of Ellipsomyxa spp., Ellipsomyxa was recovered as monophyletic. The new species is the first species of Ellipsomyxa reported from a freshwater fish in Asia and the first myxozoan reported from P. macronema.

Description, life cycle, and phylogenetics of Proterometra wigglewomble n. sp. (Digenea: Azygiidae) from the Cahaba River, Alabama, U.S.A.

We herein describe Proterometra wigglewomble n. sp. (Digenea: Azygiidae: Azygiinae) from the Cahaba River, Alabama, USA, which asexually reproduces in the compact elimia, Elimia showalteri (Lea, 1860) (Cerithioidea: Pleuroceridae) and matures in the oesophagus of the blackbanded darter, Percina nigrofasciata (Agassiz, 1854) (Perciformes: Percidae). Adults of the new species differ from congeners by having a small body and eggs having a wholly fimbriated surface that appears as a cilia-like brush border. Live naturally-shed cercariae of the new species differ from those of its congeners by having a strongly claviform tail stem bearing aspinose mammillae, a single furca, excretory pores that open on the posterior margin of the single furca, and few eggs in the cercarial distome. The behaviour of the cercaria further differentiates the new species. Naturally-shed cercariae of P. wigglewomble secrete a jelly-like adhesive that coats the surface of the furca and evidently facilitates attachment to the surface of glass, plastic, and snail shell. Attached cercariae vigorously wiggle and thrash about once attached, as if mimicking the larva of a stream insect so as to lure the blackbanded darter to eat it. Phylogenetic analyses recovered monophyletic Azygiidae, comprising monophyletic Leuceruthrinae Goldberger, 1911 and polyphyletic Azygiinae Lühe, 1909. The present study is the largest taxon sampling for Azygiidae and the first to include 28S sequences of Leuceruthrus. Compact elimia and blackbanded darter are new host records for Proterometra. The new species is the 3rd congener reported from the Cahaba River, a region renowned for its fish and snail endemic biodiversity.

Huffmanela cf. huffmani (Nematoda: Trichosomoididae) infecting swim bladder, peritoneum, and gonad of variable platyfish, Xiphophorus variatus (Cyprinodontiformes: Poeciliidae) and eastern mosquitofish, Gambusia holbrooki (Poeciliidae) in Florida; taxonomy, phylogenetic analysis, and pathological changes.

Variable platyfish, Xiphophorus variatus (Meek, 1904) (Cyprinodontiformes: Poeciliidae) and eastern mosquitofish, Gambusia holbrooki Girard, 1859 (Poeciliidae) from earthen ponds in west central Florida were examined for parasitic infections. At necropsy, we observed myriad nematodes (adults and eggs), which we identified as Huffmanela cf. huffmani, infecting the swim bladder, gonad, and visceral peritoneum. Nucleotide sequences (small subunit ribosomal DNA, 18S) of H. cf. huffmani from variable platyfish and eastern mosquitofish were identical; likewise for newly obtained 18S sequences of Huffmanela huffmani Moravec, 1987 from the swim bladder of red breast sunfish, Lepomis auritus (Linnaeus, 1758) (Centrarchiformes: Centrarchidae) and warmouth, Lepomis gulosus (Cuvier, 1829) from the San Marcos River (type locality for Huffmanela huffmani Moravec, 1987), Texas. The sequences of H. huffmani and H. cf. huffmani differed by 7 (1%) nucleotides. Pathological changes comprised proliferation of the tunica externa of the swim bladder in low-intensity infections in addition to inflammation, proliferation, and tissue necrosis of swim bladder, peritoneum, and gonad in high-intensity infections. The lesion was severe, affecting the cellular constituents of the swim bladder wall and reducing the size of the swim bladder lumen; potentially reducing swim bladder physiological efficiency. The present study is the first record of a freshwater species of Huffmanela Moravec, 1987 from beyond the San Marcos River, first record of a species of Huffmanela from a livebearer, first nucleotide sequences and phylogenetic analysis for Huffmanela, and first evidence that an infection by a species of Huffmanela causes pathological changes that could impact organ function.

Detection of Myxobolus cerebralis (Bivalvulida: Myxobolidae) in two non-Tubifex tubifex oligochaetes in the southeastern USA.

Myxobolus cerebralis (Hofer, 1903), the etiological agent of salmonid whirling disease, reportedly matures in only the oligochaete 'Tubifex tubifex'. The concept of 'T. tubifex' is problematic because it is renowned as a species complex (or having 'strains'), and many sequences ascribed to this taxon in GenBank are misidentified or indicate several cryptic species. These facts cast doubt on the long-held notion that M. cerebralis is strictly host-specific to the single definitive host, T. tubifex. Herein, as part of an ongoing regional whirling disease monitoring project, oligochaetes (452 specimens) were collected from 31 riverine sites in western North Carolina (August through September 2015) and screened for infection by M. cerebralis. The species-specific nested PCR for M. cerebralis was positive for 8 oligochaete specimens from the French Broad River Basin (Mill Creek and Watauga River) and New River Basin (Big Horse Creek). We individually barcoded these M. cerebralis-positive oligochaete specimens using cytochrome oxidase 1 (CO1) primers and then conducted a Bayesian inference phylogenetic analysis. We identified 2 oligochaete genotypes: one sister to a clade comprising Limnodrilus udekemianus (Haplotaxida: Naididae) and another sister to Limnodrilus hoffmeisteri. This is the first detection of M. cerebralis from an oligochaete in the SE USA and the first detection of M. cerebralis from an oligochaete other than T. tubifex. These results suggest that other non-T. tubifex definitive hosts can harbor the pathogen and should be considered in the context of fish hatchery biosecurity and monitoring wild trout streams for M. cerebralis and whirling disease in the southeastern USA.

Morphology, phylogenetics and pathology of "red sore disease" (coinfection by Epistylis cf. wuhanensis and Aeromonas hydrophila) on sportfishes from reservoirs in the South-Eastern United States.

The aetiological agents of red sore disease (RSD) reportedly comprise a taxonomically ambiguous stalked ciliate (a species of Epistylis) and Aeromonas hydrophila. The taxonomic identity of each pathogen remains provisional: using supra-specific morphological features for the ciliate and culture-based methods that cannot delineate bacterial strain. On 7 and 9 November 2017 and 28 May 2020, biologists and anglers reported a local epizootic (Hiwassee and Chattahoochee river basins; Georgia) wherein some moribund fish presented RSD-like lesions. The ciliates were assigned to Epistylis by morphology. The ciliate is regarded as Epistylis cf wuhanensis, as nucleotide sequences from its small subunit ribosomal DNA were identical to those of Epistylis wuhanensis. The bacterium was identified as Aeromonas hydrophila by phenotypic markers and nucleotide sequences from the DNA gyrase subunit B; our sequences comprised 3 strains and phylogenetically were recovered sister to strains of Eurasian origin. Histological sections of lesions revealed effacement or partial deterioration of the epithelium covering scales, scale loss, haemorrhaging, necrosis, oedema, and extensive inflammatory infiltrate in the dermis. This is the first nucleotide sequence information for the symbionts implicated in RSD.

Two new species of Myxobolus Bütschli, 1882 (Cnidaria: Bivalvulida: Myxobolidae) infecting the gill of the black redhorse, Moxostoma duquesnei (Lesueur) (Cypriniformes: Catostomidae) in the Little Tennessee River Basin, North Carolina.

Two new species of Myxobolus Bütschli, 1882 (Bivalvulida: Myxobolidae) are described from the gill of the black redhorse (Moxostoma duquesnei [Leueur][Cypriniformes: Catostomidae]) from the Little Tennessee River Basin, North Carolina, United States. Myxobolus branchiofilum n. sp. infects lumen of the lamellar arterioles and Myxobolus branchiopecten n. sp. infects the bone and cartilage at the tip of the gill rakers. They differ from all congeners by a combination of myxospore dimensions and the presence or absence of an iodinophilic vacuole in the sporoplasm, mucous envelope, intercapsular process, and sutural markings. A phylogenetic analysis of the small subunit ribosomal DNA recovered M. branchiopectin sister to Myxobolus sp. (AF378343) in a clade composed of 6 species of Myxobolus, which infect predominately cypriniform intermediate hosts. Myxobolus branchiofilum was recovered sister to Myxobolus ictiobus Rosser, Griffin, Quiniou, Alberson, Woodyard, Mischker, Greenway, Wise & Pote, 2016 in a clade composed of 8 species of Myxobolus, which predominately infect catostomid intermediate hosts. Histological sections of infected gill revealed intra-lamellar plasmodia of M. branchiofilum in the lumen of the lamellar arterioles and foci of M. branchiopecten developing in the bone and cartilage of the gill raker tip. These are the first myxozoans reported from the black redhorse. Given that these two new species are morphologically congeneric but recovered in distantly related clades, we discuss the persistent issue of myxobolid genera paraphyly/polyphyly.

Resurrection and emendation of Pseudoparamacroderoides Gupta & Agrawal, 1968 (Digenea: Macroderoididae), description of a new species from Vietnam, and comments on the systematics of macroderoidid genera.

We herein resurrect and emend Pseudoparamacroderoides Gupta & Agrawal, 1968 (Digenea: Macroderoididae) and describe a new species, Pseudoparamacroderoides dongthapensis n. sp., from adult specimens infecting the intestine of a riverine catfish, Mystus mysticetus Roberts, (Siluriformes: Bagridae) in the Mekong River, Vietnam. Pseudoparamacroderoides (Pseudoparamacroderoides seenghali Gupta & Agrawal, 1968 [type species]; Pseudoparamacroderoides vittati Kakaji, 1969 [= Ps. vittatusi]; Pseudoparamacroderoides raychaudhurii Agarwal & Kumar, 1983; and Pseudoparamacroderoides keni Agarwal & Agarwal, 1984) differs from other macroderoidid genera by having the combination of a subspherical oral sucker that lacks distinctly-enlarged circumoral spines; caeca that extend posteriad beyond the testes without forming a cyclocoel; testes that are approximately ≤1/3 maximum body width in diameter; a cirrus sac that is claviform, slightly dorsal to and predominantly lateral to the ventral sucker (cirrus sac partially dorsolateral to dextral or sinistral margin of ventral sucker); symmetrical vitelline fields that extend posteriad to the middle of the post-testicular space (not restricted to the inter-gonadal space) and that remain separate (not confluent) anteriorly and posteriorly; and an excretory vesicle that is I-shaped (with or without anterior swelling) and wholly post-ovarian, inter-testicular, or median to the posterior testis. Pseudoparamacroderoides dongthapensis n. sp. differs from its congeners by having an elongate hindbody (>2× forebody length) and an excretory vesicle that is approximately half as long as the body and that extends anteriad beyond the anterior testis. This is the first record of a species of Pseudoparamacroderoides from beyond the Indian sub-continent, from M. mysticetus, and from the Mekong River or from Vietnam. A diagnostic key to macroderoidid genera and a key to Pseudoparamacroderoides spp. are provided.

Prevalence, intensity and pathology of the nasal parasite Nasicola hogansi in Atlantic bluefin tuna (Thunnus thynnus).

Ectoparasitic flatworms of Nasicola (Monogenoidea: Capsalidae), which infect nasal epithelium of true tunas (Thunnus spp.), are not well studied, nor have their impacts on the host's olfactory organ been evaluated. Infections of Nasicola hogansi on Atlantic bluefin tuna, Thunnus thynnus, were investigated with emphasis on the relationship between infection prevalence, abundance and mean intensity with bluefin tuna size, sex, body condition and capture month, as well as histopathological effects. Commercially caught Atlantic bluefin tuna (n = 161, 185-305 cm curved fork length) from the Gulf of Maine were sampled during June through August 2009 for infections by N. hogansi. A total of 247 specimens of N. hogansi were collected, with a prevalence of 45.3%, mean abundance of 1.57 (CI: 1.21-2.03) and mean intensity of 3.45 (CI: 2.91-4.22). Neither fish sex nor landing month had a significant effect on parasite parameters. Larger and better-conditioned Atlantic bluefin tuna had a higher mean intensity of infection. Pathology associated with infection by N. hogansi included extensive necrosis, sloughing of the nasal epithelium and associated inflammation of underlying connective tissues. Further epidemiological and pathological study of this host-parasite system is warranted since impaired olfaction, if present, could adversely affect spawning and migration of this top ocean predator.

An updated geographic distribution of Myxobolus cerebralis (Hofer, 1903) (Bivalvulida: Myxobolidae) and the first diagnosed case of whirling disease in wild-caught trout in the south-eastern United States.

Myxobolus cerebralis (Bivalvulida: Myxobolidae), the aetiological agent of salmonid whirling disease, was detected in 2 river basins of North Carolina during 2015, which initiated the largest spatial-temporal monitoring project for the disease ever conducted within the south-eastern United States (focused mainly in eastern Tennessee and western North Carolina). A total of 2072 rainbow trout Oncorhynchus mykiss, 1,004 brown trout Salmo trutta and 468 brook trout Salvelinus fontinalis were screened from 113 localities within 7 river basins during June 2017 through October 2019. Infections were detected by pepsin-trypsin digest, microscopy and the species-specific nested polymerase chain reaction (PCR) in 19 localities across 6 river basins. Myxospore morphology was indistinguishable from the published literature. In 2019, five rainbow trout that symptomatic for whirling disease (sloping neurocranium and lordosis) were captured and processed for histopathology. Myxospores were detected in the calvarial cartilage of two deformed trout with associated erosion of the cartilage consistent with reported whirling disease lesions. This is the first report of M. cerebralis in Tennessee and the first histologically confirmed cases of whirling disease in southern Appalachian (south-eastern United States) rivers and streams and expands the distribution of M. cerebralis throughout western North Carolina and eastern Tennessee.

A new species and emendation of the seldom reported Enterohaematotrema Mehra, 1940 (Digenea: Schistosomatoidea), including a revised phylogenetic hypothesis for turtle blood flukes.

Enterohaematotrema Mehra, 1940 is emended herein based upon a review of the literature and a description of a new species (Enterohaematotrema triettruongi n. sp.) infecting yellow-headed temple turtles, Heosemys annandalii (Boulenger) (Cryptodira: Geoemydidae), in the Mekong River, Vietnam. The new species differs from the published descriptions of its congeners Enterohaematotrema palaeorticum Mehra, 1940 and Enterohaematotrema hepaticum (Simha, 1958) Simha & Chattopadhyaya, 1980 by having two distinctive oesophageal glands, a short and eversible cirrus (vs protrusive with 3 distinct processes), a dorsal common genital pore that is sinistral (vs ventral and medial), a transverse (vs longitudinal) external seminal vesicle, an oviducal seminal receptacle that is sinistral (vs dextral), and a vitellarium distributing from the caecal bifurcation (anterior to the ventral sucker) to the caecal tips (vs vitellarium not extending anteriad beyond ventral sucker in E. palaeorticum or vitellarium wholly posterior to the terminal genitalia in E. hepaticum). A phylogenetic analysis of the D1-D3 domains of the nuclear large subunit ribosomal DNA (28S) recovered Enterohaematotrema and Platt Roberts & Bullard, 2016 as sister taxa that share a recent common ancestor with the clade comprising Ruavermis Dutton & Bullard, 2020 and Coeuritrema Mehra, 1933. These flukes collectively comprise a monophyletic group of southeast Asian turtle blood flukes. This analysis also indicated that the massive, longitudinal metraterm of species of Enterohaematotrema and Uterotrema Platt & Pichelin, 1994 represents homoplasy (convergent evolution). The present study comprises the first morphological study of original specimens of any species of Enterohematotrema in more than 50 years and is the first molecular phylogenetic placement of the genus among the various turtle blood fluke lineages.

A new species of Myxobolus (Myxozoa: Bivalvulida) infecting the medulla oblongata and nerve cord of brook trout Salvelinus fontinalis in southern Appalachia (New River, NC, USA).

Myxobolus neurofontinalis n. sp. infects the brain and medulla oblongata of brook trout (Salvelinus fontinalis [Mitchill, 1814]) in the New River, western NC. It is the first species of Myxobolus described from the brook trout and resembles another congener (Myxobolus arcticus Pugachev and Khokhlov, 1979) that infects nerve tissue of chars (Salvelinus spp.). The new species differs from M. arcticus and all congeners by myxospore dimensions and by having a mucous envelope and distinctive sutural markings. A phylogenetic analysis of the small subunit rDNA (18S) suggests that the new species shares a recent common ancestor with some isolates identified as M. arcticus and that the new species and its close relatives (except Myxobolus insidiosus Wyatt and Pratt, 1973) comprise a clade of salmonid nerve-infecting myxobolids. The phylogenetic analysis indicates that several isolates of "M. arcticus" (sensu lato) in GenBank are misidentified and distantly related to other isolates taken from the type host (Oncorhynchus nerka [Walbaum, 1792]) and from nearby the type locality (Kamchatka Peninsula, Russia). Serial histological sections of infected brook trout confirmed that myxospores of the new species are intercellular and infect nerve cord and medulla oblongata only. A single infected brook trout showed an inflammatory response characterized by focal lymphocytic infiltrates and eosinophilic granulocytes; however, the remaining 4 brook trout lacked evidence of a histopathological change or demonstrable host response. These results do not support the notion that this infection is pathogenic among brook trout.

Gymnurahemecus bulbosus gen. et sp. nov. (Digenea: Aporocotylidae) infecting smooth butterfly rays, Gymnura micrura (Myliobatiformes: Gymnuridae) in the northern Gulf of Mexico, with a taxonomic key and further evidence for monophyly of chondrichthyan blood flukes.

Gymnurahemecus bulbosus gen. et sp. nov. infects the heart of smooth butterfly rays, Gymnura micrura in the Gulf of Mexico. Gymnurahemecus differs from all other accepted aporocotylid genera by having one column of C-shaped lateral tegumental spines, a medial oesophageal bulb anterior to a diverticulate region of the oesophagus, inverse U-shaped intestinal caeca, a non-looped testis, an oviducal ampulla, a Laurer's canal, and a post-caecal common genital pore. The new species, the shark blood flukes (Selachohemecus spp. and Hyperandrotrema spp.), and the chimaera blood fluke Chimaerohemecus trondheimensis are unique by having C-shaped lateral tegumental spines. Selachohemecus spp. and the new species have a single column of lateral tegumental spines, whereas Hyperandrotrema spp. and C. trondheimensis have 2-7 columns of lateral tegumental spines. The new species differs from Selachohemecus spp. most notably by having an inverse U-shaped intestine. The other ray blood flukes (Orchispirium heterovitellatum, Myliobaticola richardheardi, and Ogawaia glaucostegi) differ from the new species by lacking lateral tegumental spines, a medial oesophageal bulb, and a Laurer's canal and by having a looped testis. Phylogenetic analysis using large subunit ribosomal DNA (28S) indicated that the new species is sister to the clade that includes the other sequenced adult blood fluke (O. glaucostegi), which infects a ray in Australia. These results agree with and extend previous morphology- and nucleotide-based phylogenetic assertions that the blood flukes of early-branching jawed craniates (Chondrichthyes) are monophyletic and phylogenetically separated from the blood flukes of later-branching ray-finned fishes (Actinopterygii: Euteleostei).

Spirorchis spp. (Digenea: Schistosomatoidea) infecting map turtles (Cryptodira: Emydidae: Graptemys spp.) in southeastern North America: A new species, molecular phylogenies, and key to species.

Black-knobbed map turtles (Graptemys nigrinoda Cagle) and Alabama map turtles (Graptemys pulchra Baur) were infected with several blood flukes in Alabama (southeastern North America). Spirorchis paraminutus Roberts & Bullard n. sp. differs from its congeners by having a body that is 12-24× longer than wide, a testicular column of 10 testes that is 1/5-1/4 of the body length and located far posterior to the caecal bifurcation (the anterior-most testis is located in the posterior body half), and a common genital pore that is ventral to the ovary and 1/4-1/3 of the body length from the posterior extremity. These turtles and an Escambia map turtle (Graptemys ernsti Lovich & McCoy) were infected with Spirorchis elegans Stunkard, 1923, Spirorchis scripta Stunkard, 1923 and two innominate species of Spirorchis MacCallum, 1918. Phylogenetic analyses of the internal transcribed spacer region (ITS2) and large subunit rDNA (28S) recovered a monophyletic Spirorchis and the new species sister to Spirorchis collinsi Roberts & Bullard, 2016.

A new genus and species of turtle blood fluke (Digenea: Schistosomatoidea) from the Mekong snail-eating turtle, Malayemys subtrijuga (Schlegel & Müller) (Testudines: Geoemydidae) in Vietnam, with a reassessment of related Asiatic turtle blood flukes and molecular phylogeny.

Platt sinuosus Roberts & Bullard n. g., n. sp. (type-species) infects the kidney and mesenteric blood vessels of Mekong snail-eating turtles, Malayemys subtrijuga (Schlegel & Müller), in the Mekong River Basin. Species of Platt Roberts & Bullard n. g. are unique by the combination of having a papillate ventral sucker, vasa efferentia that are dorsal to the gonads, a massive cirrus-sac that is directed anteriad or laterad, and a vitellarium that surrounds the intestinal caeca. The new species resembles Platt ocadiae (Takeuti, 1942) Roberts & Bullard n. comb. but differs from it by having an external seminal vesicle that overlaps with or is immediately posterior to the level of the ventral sucker. Seven species previously of Hapalorhynchus Stunkard, 1922 are reassigned herein to Platt: P. odhnerensis (Mehra, 1933) Roberts & Bullard n. comb.; P. yoshidai (Ozaki, 1939) Roberts & Bullard n. comb.; P. ocadiae; P. oschmarini (Belous, 1963) Roberts & Bullard n. comb.; P. sutlejensis (Mehrotra, 1973) Roberts & Bullard n. comb.; P. synderi (Platt & Sharma, 2012) Roberts & Bullard n. comb.; and P. tkachi (Platt & Sharma, 2012) Roberts & Bullard n. comb. A dichotomous key to Platt spp. is provided. Hapalorhynchus sheilae (Mehrotra, 1973) Bourgat, 1990 and Hapalorhynchus mica (Oshmarin, 1971) Bourgat, 1990 are considered as species inquirendae, and Hapalorhynchus indicus (Thapar, 1933) Price, 1934 and Hapalorhynchus macrotesticularis (Rohde, Lee, & Lim, 1968) Brooks & Sullivan, 1981 are considered as species incertae sedis. Phylogenetic analysis of the large subunit rDNA (28S) showed P. sinuosus and P. snyderi to be sister taxa distinct from a monophyletic Hapalorhynchus and Coeuritrema platti Roberts & Bullard, 2016.

Morphological and molecular confirmation of Myxobolus cerebralis myxospores infecting wild­caught and cultured trout in North Carolina (SE USA).

We used microscopy and molecular biology to provide the first documentation of infections of Myxobolus cerebralis (Myxozoa: Myxobolidae), the etiological agent of whirling disease, in trout (Salmonidae) from North Carolina (USA) river basins. A total of 1085 rainbow trout Oncorhynchus mykiss, 696 brown trout Salmo trutta, and 319 brook trout Salvelinus fontinalis from 43 localities across 9 river basins were screened. Myxospores were observed microscopically in pepsin-trypsin digested heads of rainbow and brown trout from the Watauga River Basin. Those infections were confirmed using the prescribed nested polymerase chain reaction (PCR; 18S rDNA), which also detected infections in rainbow, brown, and brook trout from the French Broad River Basin and the Yadkin Pee-Dee River Basin. Myxospores were 9.0-10.0 µm (mean ± SD = 9.6 ± 0.4; N = 119) long, 8.0-10.0 µm (8.8 ± 0.6; 104) wide, and 6.0-7.5 µm (6.9 ± 0.5; 15) thick and had polar capsules 4.0-6.0 µm (5.0 ± 0.5; 104) long, 2.5-3.5 µm (3.1 ± 0.3; 104) wide, and with 5 or 6 polar filament coils. Myxospores from these hosts and rivers were morphologically indistinguishable and molecularly identical, indicating conspecificity, and the resulting 18S rDNA and ITS-1 sequences derived from these myxospores were 99.5-100% and 99.3-99.8% similar, respectively, to published GenBank sequences ascribed to M. cerebralis. This report comprises the first taxonomic circumscription and molecular confirmation of M. cerebralis in the southeastern USA south of Virginia.

Empruthotrema longipenis n. sp. (Monogenoidea: Monocotylidae: Merizocotylinae) from the olfactory sacs of the smooth butterfly ray Gymnura micrura (Bloch & Schneider) (Myliobatiformes: Gymnuridae) in the Gulf of Mexico.

A new species of Empruthotrema Johnston & Tiegs, 1922 is described based on specimens collected from the olfactory sacs of smooth butterfly rays Gymnura micrura (Bloch & Schneider) captured in Mobile Bay (northcentral Gulf of Mexico), Alabama, USA. Empruthotrema longipenis n. sp. is most similar to the type-species Empruthotrema raiae (MacCallum, 1916) Johnston & Tiegs, 1922 by having 12 marginal and two interhamular loculi with members of haptoral hook pair 1 located midway along the periphery of each interhamular loculus and those of hook pair 2 located at the marginal termini of the bilateral septa flanking the interhamular loculi. Empruthotrema longipenis n. sp. differs from E. raiae by having a much longer male copulatory organ and from its remaining congeners by the sinistral and extracecal ejaculatory bulb flanking the pharynx, the number of interhamular and marginal septa, and the distribution of hook pairs 1 and 2 along the haptoral margin. This is the first report of a monocotylid from the smooth butterfly ray and from Mobile Bay. The diversity of haptoral morphotypes among the currently accepted species of Empruthotrema is detailed and discussed in the context of monophyly of the genus.

Acipensericola glacialis n. sp. (Digenea: Aporocotylidae) from heart of lake sturgeon Acipenser fulvescens Rafinesque (Acipenseriformes: Acipenseridae) in the Great Lakes Basin, Lake Winnebago System, USA.

Acipensericola glacialis n. sp. infects the heart of lake sturgeon, Acipenser fulvescens (Rafinesque), in the Lake Winnebago System and differs from its only congener, Acipensericola petersoni Bullard, Snyder, Jensen & Overstreet, 2008, by having a dendritic intestine, deeply-lobed testes, a post-ovarian oötype, and a common genital pore that is medial to the dextral caecum. Acipensericola petersoni has a non-dendritic intestine, testes that are not deeply lobed, an oötype that is at level of the ovary (ventral to the ovary), and a common genital pore that is dorsal to the dextral caecum. Comparison of the large (28S) and small (18S) sub-unit ribosomal DNA and internal transcribed spacer 2 (ITS2) regions between specimens of A. glacialis n. sp. and A. petersoni revealed 13 (of 1,621 nt; 99.2% similarity in the 28S), 8 (of 1,841 nt; 99.9% similarity in the 18S), and 11 (of 442 nt; 97.5% similarity in the ITS2) nucleotide differences. Collectively, these results comprise an unexpectedly high degree of morphological and molecular similarity given the geographical (Mississippi River Basin vs Great Lakes Basin) and phylogenetic (Polyodontidae vs Acipenseridae) separation of these hosts but seemingly did not reject a previous hypothesis concerning lake sturgeon dispersal from the Mississippi Refugium following the Wisconsin glaciation ~18,000 years ago. The new species is the first nominal blood fluke described from a sturgeon.

Blood flukes of Asiatic softshell turtles: revision of Coeuritrema Mehra, 1933 (Digenea: Schistosomatoidea) and a new species infecting Chinese softshell turtles, Pelodiscus sinensis (Trionychidae), from Vietnam.

Coeuritrema Mehra, 1933, previously regarded as a junior subjective synonym of Hapalorhynchus Stunkard, 1922, herein is revised to include Coeuritrema lyssimus Mehra, 1933 (type species), Coeuritrema rugatus (Brooks et Sullivan, 1981) comb. n., and Coeuritrema platti Roberts et Bullard sp. n. These genera are morphologically similar by having a ventral sucker, non-fused caeca, two testes, a pre-testicular cirrus sac, an intertesticular ovary, and a common genital pore that opens dorsally and in the sinistral half of the body. Phylogenetic analysis of the D1-D3 domains of the nuclear large subunit ribosomal DNA (28S) suggested that Coeuritrema and Hapalorhynchus share a recent common ancestor. Coeuritrema is morphologically most easily differentiated from Hapalorhynchus by having ventrolateral tegumental papillae and a definitive metraterm that is approximately 3-7× longer than the uterus. Coeuritrema comprises species that reportedly infect Asiatic softshell turtles (Testudines: Trionychidae) only, whereas Hapalorhynchus (as currently defined) comprises blood flukes that reportedly infect those hosts plus North American musk turtles (Sternotherus Bell in Gray) and mud turtles (Kinosternon Spix), both Kinosternidae, North American snapping turtles (Chelydridae), Asiatic hard-shelled turtles (Geoemydidae) and African pleurodirans (Pelomedusidae). Coeuritrema platti sp. n. infects the blood of Chinese softshell turtles, Pelodiscus sinensis (Wiegmann), cultured in the Da Rang River Basin (Phu Yen Province, Vietnam). It differs from C. lyssimus by having a narrow hindbody (< 1.6× forebody width), ventrolateral tegumental papillae restricted to the hindbody, a short cirrus sac (< 10% of corresponding body length), a transverse ovary buttressing the caeca, a short, wholly pre-ovarian metraterm (~ 10% of corresponding body length), and a submarginal genital pore. It differs from C. rugatus by having small ventrolateral tegumental papillae, testes without deep lobes, and a Laurer's canal pore that opens posterior to the vitelline reservoir and dorsal to the oviducal seminal receptacle. The new species is only the second turtle blood fluke reported from Vietnam.