DULCITRANSVERSOTREMA N. GEN. (DIGENEA: TRANSVERSOTREMATIDAE) AND FIRST MORPHOLOGICAL AND NUCLEOTIDE-BASED CONFIRMATION OF THE LIFE CYCLE OF A TRANSVERSOTREMATID IN NORTH AMERICA.

We propose Dulcitransversotrema n. gen. for the freshwater transversotrematids Dulcitransversotrema patialense (Soparkar, 1924) n. comb. and Dulcitransversotrema chauhani (Agrawal and Singh, 1960) n. comb. Dulcitransversotrema is unique by the combination of lacking an oral sucker and an extensively coiled uterus as well as having testes that are not deeply lobed and that abut or nearly abut the inner posterolateral margins of the digestive tract (vs. deeply lobed testes that abut the cyclocoel region of the digestive tract only or that do not abut the ceca), a pre-oral genital pore (vs. post-oral genital pore), an ovary that is anterior to and abuts or nearly abuts the sinistral testis (vs. an ovary that is separated from the sinistral testis by some distance or having an ovary that is median and immediately pre-testicular), non-embryonated eggs (vs. embryonated eggs), typically having none or sparse vitelline follicles between the testes and ceca (vs. having a wholly inter-cecal vitellarium comprising a pair of highly compacted clusters of vitelline follicles anterolateral to the testes or having numerous vitelline follicles between the testes and ceca), an oblong, median, and primarily inter-testicular vitelline reservoir (vs. an extensively elongate, pre-testicular vitelline reservoir that arches anteriorly around the sinistral testis), and a subterminal excretory pore opening on the dorsal body surface between the level of the cyclocoel and posterior body end (vs. a terminal excretory pore). We describe the redia and cercaria of Dulcitransversotrema cf. patialense from the red-rimmed melania snail, Melanoides tuberculata (Müller, 1774) (Cerithioidea: Thiaridae), and their corresponding adult from beneath the scales of the zebrafish, Danio rerio (Hamilton, 1822) (Cypriniformes: Danionidae). All hosts were sampled from a spring-fed earthen pond at a private aquaculture facility near Ruskin, Florida. No transversotrematid life cycle was known from North America previously. The large subunit ribosomal DNA and ribosomal internal transcribed spacer 2 sequences of D. cf. patialense from Florida were most similar to those from D. patialense infecting red-rimmed melania from Mayagüez, Puerto Rico and differed by 134 and 69 nucleotides, respectively. Both phylogenetic analyses recovered Dulcitransversotrema as monophyletic and sister to a clade comprising Transversotrema spp. plus Crusziella formosa Cribb, Bray, and Barker, 1992.

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.

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.

RESURRECTION OF PLESIOCREADIUM WINFIELD, 1929 (DIGENEA: MACRODEROIDIDAE) WITH PHYLOGENETIC ANALYSES AND SUPPLEMENTAL OBSERVATIONS OF ITS TYPE SPECIES FROM RIVERS IN ARKANSAS, MISSISSIPPI, NEW YORK, AND TENNESSEE.

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.

ENCYCLOBREPHUS BREVIVITELLUS N. SP. (DIGENEA: PLAGIORCHIOIDEA, INCERTAE SEDIS) INFECTING THE MEKONG SNAIL-EATING TURTLE, MALAYEMYS SUBTRIJUGA, IN THE MEKONG RIVER, VIETNAM, WITH PHYLOGENETIC ANALYSES BASED ON RDNA.

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.

Plagioporus wataugaensis n. sp. (Digenea: Opecoelidae) infecting intestine of northern hogsucker, Hypentelium nigricans, and white sucker, Catostomus commersonii, (Cypriniformes: Catostomidae) from the eastern USA, including an emended diagnosis, key to Nearctic congeners, and phylogenetic analysis.

We describe a new species of Plagioporus Stafford, 1904 infecting the intestine of two catostomids in the eastern USA. We emend Plagioporus to account for Nearctic congeners having ceca terminating at the level of the testes (previously diagnosed as having ceca terminating in the post-testicular space only) and testes in the posterior body extremity (a feature not previously considered as having generic importance). Of the accepted Nearctic species, Plagioporus wataugaensis n. sp. resembles Plagioporus serotinus Stafford, 1904, Plagioporus hypentelii Hendrix, 1973, and Plagioporus hageli Fayton and Andres, 2016 but differs from them by the distribution of the vitellarium and proportional length and relative extent of the excretory vesicle. Plagioporus wataugaensis has vitelline fields that are discontinuous at the level of the ventral sucker (vs. continuous in P. serotinus and P. hypentelii) and follicles that surround the ceca (vs. wholly ventral to the ceca in P. hageli) and that span the midline dorsal to the testes (vs. slightly overlapping the lateral margins of the testes). The excretory vesicle of P. wataugaensis is wholly post-testicular and short (6-9% of the body length) (vs. reaching the level of the posterior testis, 14-24% of the body length). Phylogenetic analyses of the 28S, ITS1, 5.8S, and ITS2 rDNA recovered P. wataugaensis sister to Plagioporus sinitsini Mueller, 1934. A key to the Nearctic Plagioporus spp. is provided. We regard Plagioporus shawi (McIntosh, 1939) Margolis, 1970, Plagioporus serratus Miller, 1940, and Plagioporus loboides (Curran, Overstreet, and Tkach, 2007) Fayton and Andres, 2016 as incertae sedis.

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.

POSTHOVITELLINUM PSILOTERMINAE N. GEN., N. SP. (DIGENEA: LISSORCHIIDAE) INFECTING THE INTESTINE OF CYCLOCHEILOS ENOPLOS (CYPRINIFORMES: CYPRINIDAE) IN THE MEKONG RIVER, VIETNAM.

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.