OPECHONA CHLOROSCOMBRI AND OPECHONA CORKUMI N. SP. (DIGENEA: LEPOCREADIIDAE) FROM THE NORTHERN GULF OF MEXICO WITH PHYLOGENETIC ANALYSIS BASED ON 28S RDNA.

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.

Two new species of Unikaryon (Microsporidia) hyperparasitic in microphallid metacercariae (Digenea) from Florida intertidal crabs.

The genus Unikaryon (Microsporidia) holds exclusively hyperparasites of Platyhelminthes. Four species of Unikaryon are presently known from trematodes infecting mollusks and fish, and one from a cestode infecting a fish. Here we report two species of Unikaryon from microphallid trematode metacercariae parasitizing the brachyuran crabs, Panopeus herbstii and Pachygrapsus transversus, collected from intertidal habitats in Florida. The first microsporidium, which we assign here to a new species, Unikaryon panopei sp. n., was isolated from Microphallus sp. encysted in Panopeus herbstii from Tampa Bay. The specific designation for the second Unikaryon sp. (Unikaryon sp. 2), which occurred in metacercaria of Diacetabulum sp. found in P. transversus from the Florida Keys, is pending due to the lack of SSrDNA sequence data. Light and electron microscopy demonstrates that both species display characteristics of the genus Unikaryon including the arrangement of spores in sets of two, large posterior vacuole, and eccentric position of the polar filament. Spores of Unikaryon panopei sp. n., unlike those of Unikaryon sp. 2, assemble in large membrane-bound masses containing hundreds of organisms, and display a larger number of polar filament coils - 7-8, compared to 4-5 in Unikaryon sp. 2 The SSUrDNA-inferred phylogenetic analysis places Unikaryon panopei in one clade with Unikaryon legeri, the only other molecularly characterized member of the genus, with 94% of SSUrDNA similarity. These findings increase the number of species parasitizing trematodes and broaden the host range of Unikaryon spp.

Microsporidia Can Acquire Lamin-like Intermediate Filaments and Cell Adhesion Catenin-cadherin Complexes from the Host (?).

On their spore surfaces, Microsporidia often develop a canopy of filaments with characteristics of intermediate filaments (IF), as we demonstrated in previous studies on Thelohania sp., Ameson michaelis, and Spraguea lophii. Genomic studies indicate that among invertebrates, lamins that may localize in the cytoplasm or nucleus, are the only known IF type. These IFs can bind to the substrate containing cell adhesion molecules (CAMs) cadherins, associated with ß and γ catenins. The objects of this study were to determine whether microsporidia have CAMs with the attached IFs on their envelopes and to find out if these proteins are provided by the host. An examination was made for localization of lamins and CAMs on the spores of the mentioned above species and Anncaliia algerae, plus in the host animals. Then, we determined whether the spores of A. michaelis and A. algerae could bind vertebrate nuclear lamin onto the spore surface. We also tested transgenic Drosophila melanogaster stocks bearing cadherin-GFP to see whether developing A. algerae parasites in these hosts could acquire host CAMs. The tests were positive for all these experiments. We hypothesize that microsporidia are able to acquire host lamin IFs and cell adhesion catenin-cadherin complexes from the host.

On the systematics of some marine haploporids (Trematoda) with the description of a new species of Megasolena Linton, 1910.

Megasolena mikra sp. nov. is described from the queen angelfish, Holacanthus ciliaris (Linnaeus), off Florida, USA. The new species can be differentiated from all other species of Megasolena Linton, 1910 except Megasolena littoralis Muñoz, George-Nascimento, and Bray, 2017 in possessing testes that are smaller in diameter than the ovary. The new species can be differentiated from M. littoralis in lacking tegumental spines and possessing oral sucker papillae. Molecular data are provided for two species each of Cadenatella Dollfus, 1946, Hapladena Linton, 1910, and Megasolena Linton, 1910. Bayesian inference analysis of concatenated internal transcribed spacer region-2 (ITS2) and partial 28S rDNA sequences of 50 haploporoids revealed 1) a monophyletic Atractotrematidae Yamaguti, 1939 sister to the rest of the haploporoids tested; 2) a paraphyletic Megasoleninae Manter, 1935 - if Hapladena is included; and 3) a monophyletic Cadenatellinae Gibson and Bray, 1982 sister to the 'mugilid' haploporids. The 'mugilid' haploporids formed a monophyletic clade consisting of the subfamilies Chalcinotrematinae Overstreet and Curran, 2005, Forticulcitinae Blasco-Costa, Balbuena, Kostadinova, and Olson, 2009, Haploporinae Nicoll, 1914, and Waretrematinae Srivastava, 1937. Based on our analysis we restrict the Megasoleninae to include Megasolena, Vitellibaculum Montgomery, 1957, and Metamegasolena Yamaguti, 1970, all of which have species with two testes. To accommodate the former megasolenine taxa with a single testis, we erect the Hapladeninae subf. nov. for species in Hapladena and tentatively, Myodera Montgomery, 1957. Our results further support that haploporoids had a common marine ancestor with two testes, and that members of the Haploporoidea Nicoll, 1914 underwent diversification following a shift from a primarily marine life history with eupercarian hosts to a more euryhaline one with diadromous hosts (namely mullet).

A new microsporidium, Apotaspora heleios n. g., n. sp., from the Riverine grass shrimp Palaemonetes paludosus (Decapoda: Caridea: Palaemonidae).

We report a new microsporidium from a key species of the estuarine communities of the Gulf States, the Riverine grass shrimp, Palaemonetes paludosus. A milky-white shrimp was found in the Mobile Bay Delta, a large, oligohaline-freshwater wetland in Alabama, USA. Light microscopy of smears and thick sections of the abdominal tissues demonstrated infection with microsporidian spores enclosed in sporophorous vesicles (SVs) in sets of eight. Broadly oval spores measured 2.9 ±â€¯0.06 × 1.7 ±â€¯0.03 µm (2.5-3.3 × 1.6-1.9 µm, n = 11). SVs with a persistent membrane ranged from 4.4 to 5.6 µm in diameter. Subcuticular epithelium and underlying musculature were packed with sporonts, sporoblasts, and spores. Electron microscopy demonstrated diplokaryotic meronts that gave rise to sporont mother cells with a large single nucleus. The meront plasma membrane turned into a SV envelope, and the sporont wall segregated internally. The sporont nucleus underwent meiosis followed by two mitotic divisions accompanied by internal budding to produce four sporonts, each dividing in two uninucleate sporoblasts. Eight-spore SVs were filled with fibrillary-tubular secretions. Spores possessed 90-110-nm thick envelopes (exospore, 40-60 nm + endospore, 30-50 nm), a triangle-shaped nucleus, isofilar polar filament of 10-13 coils arranged in two-three rows, bipartite polaroplast, and a mushroom-shaped polar disk. The SSU rDNA sequence of the novel species was deposited in GenBank under Accession number MG 708238. SSU rDNA-based phylogenetic analysis indicated that the Riverine grass shrimp microsporidium was a new species and placed it in one branch with two species of Potaspora, xenoma-forming microsporidia from freshwater perciform fishes. Because morphological and developmental characters of the novel species did not fit the diagnosis of the genus Potaspora, and, based on SSU rDNA-inferred phylogenetic analyses, different host specificity, pathogenesis, and ecological considerations, we erect here the new genus Apotaspora for the Riverine grass shrimp microsporidium and name the new species Apotaspora heleios. Grouping together fish and crustacean parasites on SSU rDNA phylogenetic trees suggests that polyxenous life cycles might be a common feature of extinct and/or extant members of the studied lineage of the Microsporidia.

Two New Species of Saccocoelioides (Digenea: Haploporidae) with Phylogenetic Analysis of the Family, Including Species of Saccocoelioides from North, Middle, and South America.

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.

Description of three species of Isorchis (Digenea: Atractotrematidae) from Australia.

Three species of Isorchis Durio and Manter, 1969 are described from Australian waters. Isorchis megas sp. nov. is described from the spotbanded scat, Selenotoca multifasciata (Richardson), off Western Australia (WA) and Northern Territory (NT); Isorchis currani sp. nov. is described from S. multifasciata off NT; and Isorchis anomalus sp. nov. is described from the milkfish, Chanos chanos Forsskål, off WA. Isorchis megas sp. nov. can be differentiated from the other species of Isorchis by possessing a single, large egg that is greater than 20% of the body length; having a shorter body (the largest specimen is less than 500 µm); and utilizing a scatophagid rather than a chanid host. Isorchis currani sp. nov. can be differentiated from species of Isorchis other than I. megas sp. nov. by utilizing a scatophagid rather than a chanid host; it is differentiated from I. megas sp. nov. in having eggs that are 11-15% of the body length. Isorchis anomalus sp. nov. can be differentiated from all other species of Isorchis in possessing an irregular shaped genital pore rather than one that is circular to oblong. A Bayesian inference analysis of partial 28S rDNA sequences of the three new species of Isorchis and 30 other haploporoids revealed 1) the monophyly of the Atractotrematidae Yamaguti, 1939, 2) the two species of Isorchis infecting S. multifasciata were each other's closest relative, and 3) that Isorchis was most closely related to Pseudomegasolena Machida and Komiya, 1976 rather than Atractotrema Goto and Ozaki, 1929 although sequence data are not yet available for a member of Pseudisorchis Ahmad, 1985.

Novel Infection Site and Ecology of Cryptic Didymocystis sp. (Trematoda) in the Fish Scomberomorus maculatus.

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.

Trematode diversity in freshwater fishes of the Globe II: 'New World'.

We provide a summary overview of the diversity of trematode parasites in freshwater fishes of the 'New World', i.e. the Americas, with emphasis on adult forms. The trematode fauna of three regions, South America, Middle America, and USA and Canada (North America north of Mexico), are considered separately. In total, 462 trematode species have been reported as adults from the Americas. The proportion of host species examined for parasites varies widely across the Americas, from a high of 45% in the Mexican region of Middle America to less than 5% in South America. North and South America share no adult species, and one exclusively freshwater genus, Creptotrema Travassos, Artigas & Pereira, 1928 in the Allocreadiidae Looss, 1902 is the most widely distributed. Metacercariae of strigeiforms maturing in fish-eating birds (e.g. species of the Diplostomidae Poirier, 1886) are common and widely distributed. The review also highlights the paucity of known life-cycles. The foreseeable future of diversity studies belongs to integrative approaches and the application of molecular ecological methods. While opportunistic sampling will remain important in describing and cataloguing the trematode fauna, a better understanding of trematode diversity and biology will also depend on strategic sampling throughout the Americas.

Austrobdella cairae n. sp., an Oioxenous Marine Leech (Clitellata: Piscicolidae) from the Banded Guitarfish, Zapteryx exasperata, in the Northeastern Pacific Ocean.

A new marine leech is herein described from specimens infecting the external surfaces, including the mouth and cloaca, of the banded guitarfish, Zapteryx exasperate, captured in the Gulf of California and eastern Pacific Ocean off the coast of San Diego, California. The leech is assigned to Austrobdella by possessing continuous contractile coelomic channels that lie outside the somatic musculature along the lateral edges of the urosome (marginal lacunae), clitellar gland cells densely packed in the urosome, 5 pairs of testisacs, and 6-annulate mid-body somites. The new leech is distinguished from its 6 congeners on the basis of body size (maximum 10 mm long) and shape (sub-cylindrical trachelosome distinctly demarcated from wider urosome that is ventrally flattened, convex dorsally, and narrowing toward caudal sucker that is narrow, 20-25% of maximum body width), number of eyespots (2 pairs), shape and arrangement of the ovisacs (pyriform and limited to somites XII/XIII), and characteristics of the midgut (1 pair of mycetomes, 6 pairs of simple thin-walled crop ceca, ventral postceca wanting, and 2 pairs of dendritic diverticula emerging from anterior portion of thick-walled intestine). The new species occurs in the northeastern Pacific Ocean on a benthic elasmobranch. Examination of host specificity for each Austrobdella species using the quantitative Index of Phylogenetic Host Specificity revealed that the new species is 1 of 4 oioxenous specialists in the genus, and the remaining 3 congeners are relative generalists herein classified as euryxenous. This is the first time host specificity for members of the Piscicolidae has been quantitatively assessed. The analysis suggests that associations between marine leeches belonging in Austrobdella and their vertebrate hosts are driven by ecological influences rather than host taxonomic placement.

Two New Species of Homalometron (Digenea: Apocreadiidae) from Nearctic Freshwater Fundulids, Elucidation of the Life Cycle of H. cupuloris, and Molecular Phylogenetic Analysis of Some Congeners.

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.

Diversity, specificity and speciation in larval Diplostomidae (Platyhelminthes: Digenea) in the eyes of freshwater fish, as revealed by DNA barcodes.

Larvae (metacercariae) in some species of Diplostomidae (Platyhelminthes: Digenea) inhabit fish eyes and are difficult to identify to species based on morphology. DNA barcoding has clarified the diversity and life cycles of diplostomids in North America, Europe and Africa, but has seldom been used in parasites sampled in large numbers or at large spatial scales. Here, distance-based analysis of cytochrome c oxidase 1 barcodes and, in some specimens, internal transcribed spacer (ITS-1, 5.8S, ITS-2) sequences was performed for over 2000 diplostomids from Africa, the Middle East, Europe, Asia and the Americas. Fifty-two species of Diplostomum, Tylodelphys and Austrodiplostomum (Digenea: Diplostomidae) were distinguished. The 52 species comprise 12 identified species, six species in two species complexes and 34 putative species, and 33/52 had been delineated in previous studies. Most (23/40) of the unidentified, putative species distinguished by cytochrome c oxidase 1 distances were supported by at least one additional line of evidence. As the intensity of sampling of the 52 species increased, variation in cytochrome c oxidase 1 decreased between and increased within species, while the spatial scale at which species were sampled had no effect. Nonetheless, variation between species always exceeded variation within species. New life-cycle linkages, geographic and host records, and genetic data were recorded in several species, including Tylodelphys jenynsiae, Tylodelphys immer and Diplostomum ardeae. Species of Diplostomum inhabiting the lens are less host-specific and less numerous than those infecting other tissues, suggesting that reduced immune activity in the lens has influenced rates of speciation.

Living on the Edge: Settlement Patterns by the Symbiotic Barnacle Xenobalanus globicipitis on Small Cetaceans.

The highly specialized coronulid barnacle Xenobalanus globicipitis attaches exclusively on cetaceans worldwide, but little is known about the factors that drive the microhabitat patterns on its hosts. We investigate this issue based on data on occurrence, abundance, distribution, orientation, and size of X. globicipitis collected from 242 striped dolphins (Stenella coeruleoalba) that were stranded along the Mediterranean coast of Spain. Barnacles exclusively infested the fins, particularly along the trailing edge. Occurrence, abundance, and density of X. globicipitis were significantly higher, and barnacles were significantly larger, on the caudal fin than on the flippers and dorsal fin. Barnacles were found more frequently and in greater numbers on the dorsal rather than ventral side of the caudal fin and on the central third of dorsal and ventral fluke surfaces. Nearly all examined individuals attached with their cirral fan oriented opposite to the fluke edge. We suggest that X. globicipitis may chemically recognize dolphins as a substratum, but fins, particularly the flukes, are passively selected because of creation of vortices that increase contact of cyprids with skin and early survival of these larvae at the corresponding sites. Cyprids could actively select the trailing edge and orient with the cirri facing the main direction of flow. Attachment on the dorsal side of the flukes is likely associated with asymmetrical oscillation of the caudal fin, and the main presence on the central segment of the flukes could be related to suitable water flow conditions generated by fluke performance for both settlement and nutrient filtration.

An additional genus and two additional species of Forticulcitinae (Digenea: Haploporidae).

Forticulcita platana sp. n. and Forticulcita apiensis sp. n. are described from Mugil liza Valenciennes in Argentina, and from Mugil cephalus Linnaeus in Salt Springs, Florida, USA, respectively. Supplemental material relating to the hermaphroditic sac of Forticulcita gibsoni Blasco-Costa, Montero, Balbuena, Raga et Kostadinova, 2009 is provided from a specimen isolated from M. cephalus off Crete, Greece. Forticulcita platana can be distinguished from all species of Forticulcita Overstreet, 1982 except F. gibsoni, based on possessing small pads or gland cells along the hermaphroditic duct. It can be differentiated from that species in possessing a hermaphroditic sac that is one and a half to two times longer than wide rather than one that is approximately three times longer than wide, longer eggs (44-52 µm rather than 34-44 µm long) and a shorter post-testicular space (< 45% of the body length). Forticulcita apiensis can be differentiated from the other species of Forticulcita in possessing a testis that is shorter than or equal to the pharynx rather than one that is longer than the pharynx. Xiha gen. n. is erected for Dicrogaster fastigatus Thatcher et Sparks, 1958 as Xiha fastigata (Thatcher et Sparks, 1958) comb. n., and we tentatively consider Dicrogaster fragilis Fernández Bargiela, 1987 to be Xiha fragilis (Fernández Bargiela, 1987) comb. n. The new genus fits within the concept of Forticulcitinae Blasco-Costa, Balbuena, Kostadinova et Olson, 2009 in having a vitellarium comprised of a single elongate to subspherical mass. Xiha can be differentiated from Forticulcita in having spines lining the hermaphroditic duct, or intromittent organ. A Bayesian inference analysis of partial 28S rDNA sequences of the two New World species of Forticulcita, Xiha fastigata and previously published haploporids places Xiha fastigata within the Forticulcitinae and sister to Forticulcita. Amended diagnoses for the subfamily and for Dicrogaster Looss, 1902 are provided.

Erection of the haploporid genus Litosaccus n. g. and its phylogenetic relationship within the Haploporidae Nicoll, 1914.

Litosaccus n. g. is erected for Paralecithobotrys brisbanensis Martin, 1974 n. comb. for which an amended description is given. The new genus is morphologically similar to the haploporine Lecithobotrys Looss, 1902 but with a more elongate and cylindrical body; an infundibuliform oral sucker; a thin-walled hermaphroditic sac; a shallow genital atrium; and unequal, cylindrical and elongated caeca. It also resembles Pseudolecithobotrys Blasco-Costa, Gibson, Balbuena, Raga & Kostadinova, 2009, but the only member of that genus has a hermaphroditic sac that is twice the length of the ventral sucker, a hermaphroditic duct with intensely-staining cuboidal cells, an elongate testis, and single or paired caeca. A Bayesian inference analysis of partial 28S rDNA sequences of L. brisbanensis and 24 other haploporoids revealed that L. brisbanensis grouped with other haploporines and placed Intromugil Overstreet & Curran, 2005 in a clade with the chalcinotrematine Saccocoelioides Szidat, 1954 rather than the other seven tested waretrematine species. This analysis represents the first phylogenetic study of the Haploporidae Nicoll, 1914 that incorporates a haploporine from outside of the Mediterranean Sea.

Molecular characterization of two opecoelid trematodes from fishes in the Gulf of Mexico, with a description of a new species of Helicometra.

The plagioporine opecoelids Helicometra fasciata (Rudolphi, 1819) Odhner, 1902, and Macvicaria crassigula (Linton, 1910) Bartoli, Bray, and Gibson, 1989 have been reported from fishes in expansive geographic regions, disjointed from their type localities. New material of M. crassigula was collected from near its type locality as well as specimens resembling Helicometra fasciata sensu lato from three triglids in the Gulf of Mexico. Comparisons of the ribosomal DNA (rDNA) sequences, comprising the partial 18S rDNA, internal transcribed spacer region (= ITS1, 5.8S, and ITS2), and partial 28S rDNA gene, from M. crassigula and Helicometra fasciata sensu lato in the Gulf of Mexico were made with sequences deposited in GenBank from those species from the Mediterranean Sea. Results reveal that M. crassigula sensu stricto from the Gulf of Mexico is distinct from the two cryptic species of M. crassigula sensu lato from the Mediterranean Sea and Helicometra fasciata sensu lato in this study differs from H. fasciata sequences from the Mediterranean Sea, thus Helicometra manteri sp. nov. is described.

New genus of opecoelid trematode from Pristipomoides aquilonaris (Perciformes: Lutjanidae) and its phylogenetic affinity within the family Opecoelidae.

Bentholebouria colubrosa gen. n. et sp. n. (Digenea: Opecoelidae) is described in the wenchman, Pristipomoides aquilonaris (Goode et Bean), from the eastern Gulf of Mexico, and new combinations are proposed: Bentholebouria blatta (Bray et Justine, 2009) comb. n., Bentholebouria longisaccula (Yamaguti, 1970) comb. n., Bentholebouria rooseveltiae (Yamaguti, 1970) comb. n., and Bentholebouria ulaula (Yamaguti, 1970) comb. n. The new genus is morphologically similar to Neolebouria Gibson, 1976, but with a longer cirrus sac, entire testes, a rounded posterior margin with a cleft, and an apparent restriction to the deepwater snappers. Morphologically, the new species is closest to B. blatta from Pristipomoides argyrogrammicus (Valenciennes) off New Caledonia but can be differentiated by the nature of the internal seminal vesicle (2-6 turns or loops rather than constrictions), a longer internal seminal vesicle (occupying about 65% rather than 50% of the cirrus sac), a cirrus sac that extends further into the hindbody (averaging 136% rather than 103% of the distance from the posterior margin of the ventral sucker to the ovary), and a narrower body (27% rather than 35% mean width as % of body length). A Bayesian inference analysis of partial sequence of the 28S rDNA from Neolebouria lanceolata (Price, 1934), Cainocreadium lintoni (Siddiqi et Cable, 1960), Hamacreadium mutabile Linton, 1910, Opecoeloides fimbriatus (Linton, 1910), Podocotyloides brevis Andres et Overstreet, 2013, the new species, and previously published comparable sequences from 10 opecoelid species revealed two clades. One clade includes deep-sea (> or = 200 m) and freshwater fish opecoelids + Opecoeloides Bremser in Rudolphi, 1819, and a second clade included those opecoelids from shallow-water marine, perciform fishes.

Levisunguis subaequalis n. g., n. sp., a tongue worm (Pentastomida: Porocephalida: Sebekidae) infecting softshell turtles, Apalone spp. (Testudines: Trionychidae) in the southeastern United States.

A new tongue worm (Pentastomida) belonging to the Sebekidae Sambon, 1922 (Porocephaloidea Sambon, 1922) is described based on exemplars collected from softshell terrapins Apalone spinifera aspera (Agassiz) and Apalone ferox (Schneider) in the southeastern United States; a new genus is erected to accommodate the new species. The new species belongs in the Sebekidae because adults possess four simple hooks arranged in a trapezoid pattern on the ventral surface of the cephalothorax, a mouth opening between the anterior and posterior pairs of hooks, a terminal anus, an elongated uterus with preanal uterine pore, and a Y-shaped seminal vesicle. Nymphs possess geminate hooks, and the new species has an aquatic life-cycle in which nymphs become encapsulated in the body cavity of a freshwater fish and mature in the lungs of a terrapin. The new genus is distinct from other genera in the Sebekidae primarily by differences in hook morphology and the fact that representatives use a terrapin as a definitive host. Nymphs infecting fish and presumed to be the new species matured as postlarval juveniles conspecific with the new species when they were fed to the eastern mud turtle, Kinosternon subrubrum (Lacépède). Nymphs of the new species are anatomically similar to but larger than nymphs of Sebekia mississippiensis Overstreet, Self & Vliet, 1985 found in the mesentery of fishes captured in Florida, U.S.A. Adults of the new species differ from those of S. mississippiensis based on hook features, chloride cell pore pattern on annuli, body size, and use of a turtle rather than crocodilian definitive host. The new species is the third North American member of the Sebekidae.

Change in rank of Megaperidae (Trematoda) to Megaperinae within the Apocreadiidae and description of Haintestinum amplum n. g., n. sp.

Haintestinum amplum n. g., n. sp. is described from the scrawled cowfish, Acanthostracion quadricornis, collected in the Eastern Gulf of Mexico off Florida, USA. The new species is relatively large and shares characters of the Apocreadiidae and Megaperidae but conforms to the diagnosis of neither. It belongs in a new genus possessing a pharynx with lobed anterior margin and intestine terminating in paired ani, like in megaperids, and, when compared with apocreadiids, it shares important anatomical features, including an I-shaped excretory vesicle, canicular seminal vesicle, eye-spot remnants, and pretesticular uterus and lacks a cirrus and cirrus sac. The H-shaped intestine and large funnel-shaped oral sucker without a U-shaped sphincter encircling half the anterior aperture are the most notable diagnostic characters of the new monotypic genus. Additionally, the phylogenetic position of the Megaperidae is investigated for the first time, using analysis of partial 28S rDNA gene sequences from H. amplum, two species in the Megapera, Thysanopharynx elongatus, and previously published 28S sequences of species from members of the Apocreadiata, Haploporoidae, Lepocreadiata, and Opisthorchiata. The resulting analysis demonstrated a close relationship among the new genus and the three species of megaperids, and the megaperids were most closely allied with Schistorchis zancli of the apocreadiids. Moreover, we now consider Megaperidae as the subfamily Megaperinae within the Apocreadiidae.