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).

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.

Huffmanela markgracei sp. n. (Nematoda: Trichosomoididae) from buccal cavity of Atlantic sharpnose shark, Rhizoprionodon terraenovae (Carcharhiniformes: Carcharhinidae), in the northwestern Gulf of Mexico off Texas.

Eggs of Huffmanela markgracei sp. n. infected one of three Atlantic sharpnose sharks, Rhizoprionodon terraenovae (Richardson) (Carcharhiniformes: Carcharhinidae) captured by bottom long-line in the northwestern Gulf of Mexico off Padre Island, Texas. Eggs in the skin formed sinuous tracks (1-8 eggs wide; 1-4 eggs deep; 150 eggs/mm2) occupying a swath of the skin 22 cm x 2 cm on the tongue, branchial arches and the dorsal surface of the buccal cavity. Eggs had transverse eggshell ridges (branching and non-branching), had shells that were clear, amber or brown, and measured 90-113 microm (x = 102 +/- 4; n = 190) long, 38-54 microm (43 +/- 3; 190) wide, 3-5 microm (4 +/- 0; 190) in eggshell thickness with protruding polar plugs 8-12 microm (10 +/- 1; 190) wide. Apparently fully developed larvae in eggs were 255-335 microm (299 +/- 26; 30) long, 8-10 microm (9 +/- 1; 30) wide, and in-folded 5-6 (6 +/- 0; 30) times. Some of these larvae were emerging from eggs in the skin. The new species differs from congeners by the combination of having a large, spindle-shaped egg, transverse eggshell ridges, an envelope that is smooth, tightly-apposed to the eggshell and surrounds the entire eggshell inclusive of the polar plugs, and a large larva. This is the first report of a species of Huffmanela Moravec, 1987 from a chondrichthyan in the Gulf of Mexico and from a shark not assigned to Carcharhinus.

A New Species of Neoalbionella (Copepoda: Siphonostomatoida: Lernaeopodidae) From Skin of the Gulper Shark, Centrophorus granulosus (Squaliformes: Centrophoridae) In the Northern Gulf of Mexico.

Using light and scanning electron microscopy of male and female copepods, we herein describe a new species of Neoalbionella Özdikmen, 2008 (Copepoda: Lernaeopodidae), Neoalbionella benzipirata n. sp., infecting the skin of the gulper shark, Centrophorus granulosus (Bloch and Schneider, 1801), in the northern Gulf of Mexico. Females of the new species were assigned to Neoalbionella by having 3 setae on the maxillule lateral palp (vs. 2 setae in Lernaeopoda Nordmann, 1832) and a single accessory denticle on the maxilliped subchela claw (vs. ≥2 denticles in Lernaeopoda). Females of the new species resemble those of Neoalbionella longicaudata ( Hansen, 1923 ) Özdikmen, 2008 but can be differentiated from them and those of all congeners by having a unique configuration of 16 dorsocephalic sensilla; an antennule terminal segment having 6 apical setae comprising tubercles 1 and 3, digitiform seta 4, 2 subequal setae of complex 5, and flagelliform seta 6; an antenna exopod with a thickened dorsal ridge, smooth (convex) outer surface with 2 short papillae, and rugose (concave) inner surface plus a terminal endopodal segment having a large dorsal hook, medial spine, bifid distal tubercle, and spinulose ventral process; a mandible having the formula P1, S1, P1, S1, P1, S1, B5; and a single pair of anterolateral spines on the mouth cone labium. Males attached to the posteroventral surface of the females' genital trunk were assigned to Neoalbionella by having tapered (not inflated) caudal rami directed posteroventrally. They were differentiated from previously reported congeneric males by having an antennule with the same number and configuration of apical setae as the female; an antenna with a spatulate exopod having minute spinules along its apex plus a terminal endopodal segment with components the same as the female; a mandible with formula P2, S1, P1, S1, B5; a labium with 1 pair of anterolateral spines like the female; and a conspicuous mediative process with bifid tip. Detail of the dorsocephalic shield (in females) and mouth cone labium (in both sexes) previously had not been used to diagnose Neoalbionella spp. We also dissected voucher specimens of " Neoalbionella longicaudata" (collected from gulper sharks in the western Pacific Ocean off Japan by Shiino in 1956), which we morphologically diagnosed as an innominate species, Neoalbionella sp. This is the first report of a species of Neoalbionella from the Gulf of Mexico and the second species of Neoalbionella reported from C. granulosus, a deepwater shark seldom encountered and surveyed for parasitology.

Neotropical Turtle Blood Flukes: Two New Genera and Species from the Amazon River Basin with a Key to Genera and Comments on a Marine-Derived Parasite Lineage in South America.

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.

Exotic "Gill Lice" Species (Copepoda: Lernaeopodidae: Salmincola SPP.) Infect Rainbow Trout (Oncorhynchus mykiss) and Brook Trout (Salvelinus fontinalis) in the Southeastern United States.

Salmincola californiensis infected 25 of 31 (prevalence 0.8; intensity 2-35 [mean 6.6 ± standard deviation 7.7; n = 25]) rainbow trout, Oncorhynchus mykiss, from a private trout farm connected to the Watauga River, North Carolina. Salmincola edwardsii infected all of 9 (1.0; 2-43 [9.3 ± 13.0; 9]) brook trout, Salvelinus fontinalis, from Big Norton Prong, a tributary of the Little Tennessee River, North Carolina. Both lernaeopodids are well-known salmonid pathogens, but neither is native to, nor has been previously taxonomically confirmed from, the southeastern United States. Herein, we (1) use light and scanning electron microscopy to identify and provide supplemental morphological observations of these lernaeopodids, (2) furnish complementary molecular sequence data from the 28S rDNA (28S), and (3) document the pathological effects of gill infections. We identified and differentiated these lernaeopodids by the second antenna (exopod tip with large [S. californiensis] vs. slender [S. edwardsii] spines; endopod terminal segment with subequal ventral processes shorter than [S. californiensis] vs. longer than or equal to [S. edwardsii] dorsal hook), maxilliped palp (length typically ≤1/3 [S. californiensis] vs. 1/3-1/2 [S. edwardsii] subchela length exclusive of claw), and bulla (sub-circular and concave on manubrium's side [S. californiensis] vs. non-stellate [S. edwardsii]). Analysis of the 28S rDNA sequences confirmed our taxonomic assignments as demonstrated by 100% sequence similarity among the sympatric, morphologically-conspecific isolates. Histopathology revealed focal gill epithelial hyperplasia, obstruction of interlamellar water channels, lamellar fusion, and crypting of gill filaments. High intensity infections by either lernaeopodid are surveillance-worthy because they are potentially pathogenic to trout in the southeastern United States.

A new species of Trichosomoididae (Nematoda) from skin of red snapper, Lutjanus campechanus (Perciformes: Lutjanidae), on the Texas-Louisiana shelf, northern Gulf of Mexico.

Eggs and larvae of Huffmanela oleumimica n. sp. infect red snapper, Lutjanus campechanus (Poey, 1860), were collected from the Texas-Louisiana Shelf (28°16'36.58″N, 93°03'51.08″W) and are herein described using light and scanning electron microscopy. Eggs in skin comprised fields (1-5 × 1-12 mm; 250 eggs/mm(2)) of variously oriented eggs deposited in dense patches or in scribble-like tracks. Eggs had clear (larvae indistinct, principally vitelline material), amber (developing larvae present) or brown (fully developed larvae present; little, or no, vitelline material) shells and measured 46-54 µm (x = 50; SD ± 1.6; n = 213) long, 23-33 (27 ± 1.4; 213) wide, 2-3 (3 ± 0.5; 213) in eggshell thickness, 18-25 (21 ± 1.1; 213) in vitelline mass width, and 36-42 (39 ± 1.1; 213) in vitelline mass length with protruding polar plugs 5-9 (7 ± 0.6; 213) long and 5-8 (6 ± 0.5; 213) wide. Fully developed larvae were 160-201 (176 ± 7.9) long and 7-8 (7 ± 0.5) wide, had transverse cuticular ridges, and were emerging from some eggs within and beneath epidermis. The new species differs from its congeners by having eggs <65 µm in total length and that have a brown eggshell when fully developed, an envelope throughout development, and irregularly-dispersed eggshell spines plus a larva >110 µm long with transverse cuticular ridges. The eggs lack a spindle-shaped envelope, polar filaments, and eggshell ridges. This is the first report of a species of Huffmanela from a snapper (Lutjanidae) or from the Gulf of Mexico. A table of egg and larval characteristics, hosts, and localities for Huffmanela spp. is provided.

Blood flukes (Digenea: Aporocotylidae) of epipelagic lamniforms: redescription of Hyperandrotrema cetorhini from basking shark (Cetorhinus maximus) and description of a new congener from shortfin mako shark (Isurus oxyrinchus) off Alabama.

We emend the original generic diagnosis for Hyperandrotrema Maillard and Ktari, 1978 , and redescribe its type species Hyperandrotrema cetorhini Maillard and Ktari, 1978 (Digenea: Aporocotylidae Odhner, 1912), based on the holotype and 2 paratypes collected from the heart of basking shark (Cetorhinus maximus). We also describe Hyperandrotrema walterboegeri Orélis-Ribeiro and Bullard n. sp. based on light and scanning electron microscopy of 6 adult specimens collected from the heart of a shortfin mako shark (Isurus oxyrinchus Rafinesque, 1810) captured from Viosca Knoll (29°11.70'N, 88°33.32'W; 123 km southwest of Dauphin Island, Alabama), northern Gulf of Mexico. Hyperandrotrema spp. infect lamniforms and differ from all other nominal aporocotylids at least by having a ventrolateral field of robust C-shaped spines (rather than transverse rows of minute, shaft-like spines), an inverse U-shaped intestine with extremely elongate ceca terminating near the level of the excretory bladder, and a common genital pore that comprises the dorsal opening of a common genital atrium. Adults of the new species exceeded 12 mm in total length, making them the largest of the nominal fish blood flukes. The new species further differs from H. cetorhini by the combination of having an adult body that is 7-8 times longer than wide, large midbody tegumental spines measuring 25-38 µm long × 10-12 µm wide, a long vas deferens 4-5% of the body length, a testis 9-11 times longer than wide, and a large ootype 105-150 µm long × 85-105 µm wide. This is the first report of Hyperandrotrema from the Gulf of Mexico and the second aporocotylid species reported from an epipelagic elasmobranch. Our results demonstrate that ecologically related (epipelagic, marine) and phylogenetically related (Lamniformes) definitive hosts are infected by morphologically similar (congeneric) fish blood flukes.

Huffmanela cf. carcharhini (Nematoda: Trichosomoididae: Huffmanelinae) from skin of a sandbar shark, Carcharhinus plumbeus, in the Pacific Ocean.

Eggs of Huffmanela cf. carcharhini from the skin of an aquarium-held, juvenile sandbar shark, Carcharhinus plumbeus , from the Pacific Ocean were studied using light and scanning electron microscopy. Grossly, eggs imparted a scribble-like skin marking approximately 130 × 60 mm on the right side of the shark's snout adjacent to its eye and nostril. Fresh (unfixed) eggs were elliptical, 75-95 µm long (x¯  =  85 µm, SD  =  ±4.5; n  =  75), 48-63 µm wide (53 ± 3.4; 75), 8-10 µm in shell thickness (9 ± 1.3; 27), 45-68 µm in vitelline mass length (52 ± 6.9; 8); had a smooth shell surface and nonprotruding polar plugs 8-13 µm wide (10 ± 1.5; 73); lacked thin filaments, superficial envelope, and shell spines; sank in 35 ppt artificial seawater; and did not spontaneously hatch after 12 hr in 35 ppt artificial seawater. Formalin-fixed eggs measured 193 days postfixation were 75-95 µm long (84 ± 3.9; 150), 45-60 µm wide (50 ± 2.2; 150), 5-10 µm in shell thickness (8 ± 1.2; 87), 45-60 µm in vitelline mass length (51 ± 3.0; 92), and 30-40 µm in vitelline mass width (33 ± 2.0; 84), and had nonprotruding polar plugs that were 10-15 µm long (11 ± 1.4; 93) and 8-10 µm wide (9 ± 1.1; 108). Forcibly hatched first-stage larvae (unfixed) were filiform, 188-273 µm long (212 ± 25.5; 13), 8-13 µm wide (10 ± 1.2; 13), and had fine transverse striations. Eggs infected the epidermis only. Histology revealed intra-epithelial inflammation with eosinophilic granulocytes and hyperplasia, plus dermal lymphofollicular hyperplasia associated with the infection. The eggs of H. cf. carcharhini likely undergo considerable ex utero development before being sloughed (unhatched) from the host, along with epidermal cells.