AZYGIID PARASITES OF NORTH AMERICAN ENDEMIC PLEUROCERIDS AND CENTRARCHIDS: REVISION OF LEUCERUTHRUS MARSHALL AND GILBERT, 1905 (DIGENEA: AZYGIIDAE), DESCRIPTION OF TWO NEW SPECIES, AND PHYLOGENETIC ANALYSIS.

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no spines on the tail stem, distinct anterior and posterior ridges that flank the tail stem portion accommodating the withdrawn distome, minute protuberances occupying the lateral margin of the tail stem for its entire length, and protuberances that encircle the anterior third of the posterior tail stem (immediately posterior to the tail stem portion containing the withdrawn distome).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

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.

New species of Proterometra (Digenea: Azygiidae) and its life cycle in the Chickasawhay River, Mississippi, USA, with supplemental observations of Proterometra autraini.

We describe Proterometra ariasae n. sp. based upon cercariae shed from a freshwater snail, Pleurocera sp., and adults infecting the buccal cavity of longear sunfish, Lepomis megalotis, captured from the Chickasawhay River, Mississippi, USA. We also provide supplemental observations of cercarial and adult paratypes of Proterometra autraini from the Au Train River, Michigan, USA. Sequence data for the ribosomal internal transcribed spacer 2 (ITS2) from adults and cercariae of the new species were identical. Adults of the new species differ from congeners by having (i) a markedly large body, (ii) a proportionally large oral sucker, (iii) ovoid testes, (iv) a strongly muscular and laterally expanded pars prostatica, (v) a uterus that is extensively convoluted between the ovary and ventral sucker (vi) and a vitellarium as long as the caeca and extending posteriad beyond the anterior margin of the testes. Cercariae of the new species differ from those of its congeners by having (i) a tail stem that is shorter than 10mm and that lacks a medial constriction, (ii) obcordate furcae that are wider than long, (iii) mamillae distributed throughout the anterior tail stem only, and (v) a proportionally small distome that has relatively few uterine eggs and remains withdrawn in the anterior tail stem region in actively swimming cercariae. This is the first report of Proterometra from Mississippi, the second description to employ morphology and sequence data to elucidate a life cycle for Proterometra, and the third species of Proterometra from an intermediate host not assigned to Elimia.

First Record of Transversotrema Witenberg, 1944 (Digenea) from the Americas, with Comments on the Taxonomy of Transversotrema patialense (Soparkar, 1924) Crusz and Sathananthan, 1960, and an Updated List of Its Hosts and Geographic Distribution.

Specimens of Transversotrema patialense (sensu lato) ( Soparkar, 1924 ) Crusz and Sathananthan, 1960 (Digenea: Transversotrematidae) infected the skin (epidermal spaces beneath scales near pectoral fins) of 4 of 126 (prevalence 3%; mean intensity 1.8) zebrafish ( Danio rerio (Hamilton, 1822) [Cypriniformes: Cyprinidae]) purchased in 2009 and cultured by a California (USA) fish supplier. These fish were sold as "laboratory-reared" and "specific pathogen free," purportedly raised in a recirculating aquaculture system that included zebrafish only. We herein describe the morphological features of this transversotrematid using light and scanning electron microscopy, provide a comprehensive list of hosts (snails and fishes) and geographic locality records for specimens reported as T. patialense, which is perhaps a species complex, and provide a brief historical synopsis of the taxonomic and life history research that has been conducted on this fluke. No species of Transversotrema previously had been reported from the Americas; however, this discovery is not surprising given that: (1) a suitable intermediate host (red-rimmed melania, Melanoides tuberculata (Müller, 1774) [Cerithioidea: Thiaridae]) has been established in California and elsewhere in North America, (2) the zebrafish is a susceptible definitive host, and (3) T. patialense reportedly matures on a broad ecological and phylogenetic spectrum of freshwater fishes. To our knowledge, this is the northern-most geographic locality record for a species of this genus. We suspect this case study represents an example of a parasite that may now be established in North America by the fortuitous co-occurrence of a susceptible, exotic snail host (the red-rimmed melania) and a susceptible, widely distributed, exotic fish host (the zebrafish).

Skin lesions on yellowfin tuna Thunnus albacares from Gulf of Mexico outer continental shelf: Morphological, molecular, and histological diagnosis of infection by a capsalid monogenoid.

We characterize lesion-associated capsaline infections on yellowfin tuna, Thunnus albacares, in the Gulf of Mexico by comparing our specimens with published descriptions and museum specimens ascribed to Capsala biparasiticum and its synonyms: vouchers of C. biparasiticum from parasitic copepods; the holotype of Capsala neothunni; and vouchers of Capsala abidjani. Those from parasitic copepods differed by having a small, rounded body, large anterior attachment organs, closely spaced dorsomarginal body sclerites, small testes, and a short and wide testicular field. No morphometric feature in the holotype of C. neothunni ranged outside of that reported for the newly-collected specimens, indicating conspecificity of our specimens. The specimens of C. abidjani differed by having a large anterior attachment organ, few and dendritic testes, and a short, wide testicular field. Large subunit ribosomal DNA (28S) sequences grouped our specimens and Capsala sp. as sister taxa and indicated a phylogenetic affinity of Nasicola klawei. The haptoral attachment site comprised a crater-like depression surrounded by a blackish-colored halo of extensively rugose skin, with abundant pockmarked-like, irregularly-shaped oblong or semi-circular epidermal pits surrounding these attachment sites. Histology confirmed extensive folding of epidermis and underlying stratum laxum, likely epidermal hyperplasia, foci of weak cell-to-cell adhesions among apical malpighian cells as well as that between stratum germinativum and stratum laxum, myriad goblet cells in epidermis, rodlet cells in apical layer of epidermis, and lymphocytic infiltrates and melanin in dermis. The present study comprises (i) the first published report of this parasite from yellowfin tuna captured in the Gulf of Mexico-NW Atlantic Ocean Basin, (ii) confirmation of its infection on the skin (rather than on a parasitic copepod), (iii) the first molecular data for this capsaline, and (iv) the first observations of histopathological changes associated with a capsalid infection on a wild-caught epipelagic fish.

Community Structure of Skin Microbiome of Gulf Killifish, Fundulus grandis, Is Driven by Seasonality and Not Exposure to Oiled Sediments in a Louisiana Salt Marsh.

Mucus of fish skin harbors complex bacterial communities that likely contribute to fish homeostasis. When the equilibrium between the host and its external bacterial symbionts is disrupted, bacterial diversity decreases while opportunistic pathogen prevalence increases, making the onset of pathogenic bacterial infection more likely. Because of that relationship, documenting temporal and spatial microbial community changes may be predictive of fish health status. The 2010 Deepwater Horizon oil spill was a potential stressor to the Gulf of Mexico's coastal ecosystem. Ribosomal intergenic spacer analysis (RISA) and pyrosequencing were used to analyze the bacterial communities (microbiome) associated with the skin and mucus of Gulf killifish (Fundulus grandis) that were collected from oiled and non-oiled salt marsh sites in Barataria Bay, LA. Water samples and fin clips were collected to examine microbiome structure. The microbiome of Gulf killifish was significantly different from that of the surrounding water, mainly attributable to shifts in abundances of Cyanobacteria and Proteobacteria. The Gulf killifish's microbiome was dominated by Gammaproteobacteria, specifically members of Pseudomonas. No significant difference was found between microbiomes of fish collected from oiled and non-oiled sites suggesting little impact of oil contamination on fish bacterial assemblages. Conversely, seasonality significantly influenced microbiome structure. Overall, the high similarity observed between the microbiomes of individual fish observed during this study posits that skin and mucus of Gulf killifish have a resilient core microbiome.

Proterometra epholkos sp. n. (Digenea: Azygiidae) from Terrapin Creek, Alabama, USA: molecular characterization of life cycle, redescription of Proterometra albacauda, and updated lists of host and geographic locality records for Proterometra spp. in North America.

Proterometra epholkos sp. n. asexually reproduces in the stream dwelling prosobranch, Elimia cf. modesta (Cerithioidea: Pleuroceridae) and infects the buccal cavity epithelium of spotted bass, Micropterus punctulatus (Perciformes: Centrarchidae) in the Coosa River (Terrapin Creek; N33°51'36.56″, W85°31'28.15″; Cleburne County, Alabama, USA). We characterize cercariae and adults of the new species using morphology and molecular sequence data and redescribe its morphologically similar congener Proterometra albacauda based on the holotype and paratype (USNPC Nos. 61229-30). The new species can be distinguished most easily from P. albacauda by the combination of having cercariae with long mamillae (>100µm) that encircle the tail stem anteriorly, that are restricted to 1 lateral column per body margin at midbody, and that are absent from the medial surface of the tail stem as well as by having adults with a partly extracecal uterus, a transverse metraterm occupying the space between the oral sucker and prostatic sac, and a vitellarium that is longer than the ceca and extends anteriad to the level of or beyond the posterior margin of the oral sucker. Sequence data from the ribosomal internal transcribed spacer 2 (ITS2; 251bp) did not reject the notion that the cercariae and adults we collected simultaneously from those infected, sympatric, individual snails and fish in Terrapin Creek were conspecific. Also provided herein for species of Proterometra are (i) taxonomic keys for cercariae and adults based on morphological and behavioral characteristics sourced from the published literature, (ii) updated lists of host records (prosobranchs and fishes) and geographic locality records for Proterometra spp., and (iii) synopses and assessments of the morphological features previously used to differentiate them. Proterometra macrostoma (type species), Proterometra melanophora, and Proterometra hodgesiana are species inquirendae; requiring new collections from type localities and hosts concomitant with neotype designations. P. macrostoma seems a repository for conspicuous, furcocystocercous cercariae shed from freshwater prosobranchs in eastern North American rivers and streams. The specific epithet "pinguis" associated with specimens purportedly infecting Esox lucius and deposited by JF Mueller is a nomen nudum. Proterometra guangzhouensis, Proterometra sillagae, Proterometra brachyuran, and Proterometra lamellorchis are incertae sedis. Significant barriers to characterizing biodiversity and distributions (host range and geographic distribution) of Proterometra spp. comprise a paucity of data on adult morphology, dubious species-level identification or a lack of information regarding prosobranch hosts, lack of molecular data for putative comparisons among fluke 'strains' and species as well as between cercariae and adults, lack of consistency in terminology, and indeterminate homology for key morphological features. Uncertainty about the providence and identity of, or absence of, accessioned museum materials of P. macrostoma, Proterometra catenaria, and P. hodgesiana together represent another fundamental problem. The present study comprises the first description of a new species of Proterometra in nearly 20years, first report of a species of the genus from the Coosa River (Mobile-Tensaw River Basin) and from these host species, and first use of molecular sequence data to elucidate a life cycle for a species of Proterometra.

Simultaneous activation of multiple signal transduction pathways confers poor prognosis in acute myelogenous leukemia.

Deregulation of signal transduction pathways (STPs) may promote leukemogenesis by conferring cell proliferation and survival advantages in acute myelogenous leukemia (AML). Several agents targeting STPs are under development; however, redundancy and cross-talk between STPs could activate multiple downstream effectors and this could negate the effect of single-target inhibition. The frequency of concurrent activation of multiple STPs in AML and the prognostic relevance of STP activation in AML are unknown. STP protein expression (PKCalpha, ERK2, pERK2, AKT, and pAKT) was measured by Western blot in samples from 188 patients with newly diagnosed, untreated AML. In univariate and multivariate analysis high levels of PKCalpha, ERK, pERK, and pAKT, but not AKT, were adverse factors for survival as was the combination variable PKCalpha-ERK2&pERK2-pAKT. Survival progressively decreased as the number of activated pathways increased. Patients were more likely to have none or all 3 pathways activated than was predicted based on the frequency of individual pathway activation, strongly suggesting that cross-activation occurred. Simultaneous activation of multiple STPs is common in AML and has a progressively worse adverse effect on prognosis. It is thus likely that only combinations of agents that target the multiply activated STPs will be beneficial for patients with AML.

Comparison of the five-electrode-derived EASI electrocardiogram to the Mason Likar electrocardiogram in the prehospital setting.

This study compared the 5-electrode-derived EASI electrocardiogram (ECG) with the conventional Mason-Likar ECG in 200 consecutive patients with chest pain transported to 3 hospitals by 2 different emergency medical services. No significant differences were observed between the 2 systems for the detection of relevant electrocardiographic abnormalities. A questionnaire administered to participating emergency medical personnel revealed a high degree of acceptability of the EASI ECG, with some participants commenting that the sternal and mid-axillary locations of the EASI electrodes made them easier to apply, especially to women, than conventional precordial electrodes.