Replacing mechanical protection with colorful faces-twice: parallel evolution of the non-operculate marine worm-snail genera Thylacodes (Guettard, 1770) and Cayo n. gen. (Gastropoda: Vermetidae).

Vermetid worm-snails are sessile and irregularly coiled marine mollusks common in warmer nearshore and coral reef environments that are subject to high predation pressures by fish. Often cryptic, some have evolved sturdy shells or long columellar muscles allowing quick withdrawal into better protected parts of the shell tube, and most have variously developed opercula that protect and seal the shell aperture trapdoor-like. Members of Thylacodes (previously: Serpulorbis) lack such opercular protection. Its species often show polychromatic head-foot coloration, and some have aposematic coloration likely directed at fish predators. A new polychromatic species, Thylacodes bermudensis n. sp., is described from Bermuda and compared morphologically and by DNA barcode markers to the likewise polychromatic western Atlantic species T. decussatus (Gmelin, 1791). Operculum loss, previously assumed to be an autapomorphy of Thylacodes, is shown to have occurred convergently in a second clade of the family, for which a new genus Cayo n. gen. and four new western Atlantic species are introduced: C. margarita n. sp. (type species; with type locality in the Florida Keys), C. galbinus n. sp., C. refulgens n. sp., and C. brunneimaculatus n. sp. (the last three with type locality in the Belizean reef) (all new taxa authored by Bieler, Collins, Golding & Rawlings). Cayo n. gen. differs from Thylacodes in morphology (e.g., a protoconch that is wider than tall), behavior (including deep shell entrenchment into the substratum), reproductive biology (fewer egg capsules and eggs per female; an obliquely attached egg capsule stalk), and in some species, a luminous, "neon-like", head-foot coloration. Comparative investigation of the eusperm and parasperm ultrastructure also revealed differences, with a laterally flattened eusperm acrosome observed in two species of Cayo n. gen. and a spiral keel on the eusperm nucleus in one, the latter feature currently unique within the family. A molecular phylogenetic analysis based on mitochondrial and nuclear rRNA gene sequences (12SrRNA, trnV, 16SrRNA, 28SrRNA) strongly supports the independent evolution of the two non-operculate lineages of vermetids. Thylacodes forms a sister grouping to a clade comprising Petaloconchus, Eualetes, and Cupolaconcha, whereas Cayo n. gen is strongly allied with the small-operculate species Vermetus triquetrus and V. bieleri. COI barcode markers provide support for the species-level status of the new taxa. Aspects of predator avoidance/deterrence are discussed for these non-operculate vermetids, which appear to involve warning coloration, aggressive behavior when approached by fish, and deployment of mucous feeding nets that have been shown, for one vermetid in a prior study, to contain bioactive metabolites avoided by fish. As such, non-operculate vermetids show characteristics similar to nudibranch slugs for which the evolution of warning coloration and chemical defenses has been explored previously.

Molecular and morphological evidence for nine species in North American Australapatemon (Sudarikov, 1959): a phylogeny expansion with description of the zygocercous Australapatemon mclaughlini n. sp.

Zygocercous (aggregating) cercarial larvae were recently discovered emerging from a physid snail during a molecular survey of cercariae from molluscs in lakes in central Alberta, Canada. This manuscript delves into the characterization of these cercariae through morphological and molecular techniques and provides the first genetic information for a zygocercous larval trematode. Analyses of cytochrome c oxidase I of mitochondrial DNA and two partial regions of nuclear ribosomal DNA sequences revealed the zygocercous cercariae to belong to the genus Australapatemon Sudarikov, 1959. Further analyses of sequences of Australapatemon burti (Miller, 1923), from cercariae and adults collected from across North America, indicate a complex of nine genetically-distinct lineages within this species, a surprising level of diversity. The zygocercous cercariae, along with adult worms collected from ducks in Manitoba, Canada, and from Mexico, represent one of these lineages, and are herein described as Australapatemon mclaughlini n. sp. Seven lineages cannot yet be identified, but one is tentatively identified as Australapatemon burti.

Non-native molluscan colonizers on deliberately placed shipwrecks in the Florida Keys, with description of a new species of potentially invasive worm-snail (Gastropoda: Vermetidae).

Artificial reefs created by deliberately sinking ships off the coast of the Florida Keys island chain are providing new habitat for marine invertebrates. This newly developing fouling community includes the previously reported invasive orange tube coral Tubastraea coccinea and the non-native giant foam oyster Hyotissa hyotis. New SCUBA-based surveys involving five shipwrecks spanning the upper, middle, and lower Florida Keys, show T. coccinea now also established in the lower Keys and H. hyotis likewise extending to new sites. Two additional mollusks found on the artificial reefs, the amathinid gastropod Cyclothyca pacei and gryphaeid oyster Hyotissa mcgintyi, the latter also common in the natural reef areas, are discussed as potentially non-native. A new species of sessile, suspension-feeding, worm-snail, Thylacodes vandyensis Bieler, Rawlings & Collins n. sp. (Vermetidae), is described from the wreck of the USNS Vandenberg off Key West and discussed as potentially invasive. This new species is compared morphologically and by DNA barcode markers to other known members of the genus, and may be a recent arrival from the Pacific Ocean. Thylacodes vandyensis is polychromatic, with individuals varying in both overall head-foot coloration and mantle margin color pattern. Females brood stalked egg capsules attached to their shell within the confines of their mantle cavity, and give rise to crawl-away juveniles. Such direct-developing species have the demonstrated capacity for colonizing habitats isolated far from their native ranges and establishing rapidly growing founder populations. Vermetid gastropods are common components of the marine fouling community in warm temperate and tropical waters and, as such, have been tagged as potentially invasive or with a high potential to be invasive in the Pacific Ocean. As vermetids can influence coral growth/composition in the Pacific and have been reported serving as intermediate hosts for blood flukes of loggerhead turtles, such new arrivals in the Florida Keys National Marine Sanctuary are of concern. Growing evidence indicates that artificial reefs can act as permanent way-stations for arriving non-natives, providing nurseries within which populations may grow in an environment with reduced competition compared to native habitats. Consequently, artificial reefs can act as sentinels for the appearance of new species. Ongoing monitoring of the developing molluscan fauna on the artificial reefs of the Florida Keys is necessary to recognize new invasions and identify potential eradication targets, thereby assuring the health of the nearby natural barrier reef.

Epidemiological study of dogs with otitis externa in Cape Breton, Nova Scotia.

From May 2008 to December 2013, 320 cases of otitis externa were diagnosed among 2012 dogs undergoing routine physical examinations at Celtic Creatures Veterinary Clinic, Sydney River, Nova Scotia for a diagnosis frequency of 15.9% [95% confidence interval (CI): 14.3% to 17.6%]. Twenty-four percent of these dogs exhibited 1 or multiple recurrences despite initial treatment with topical antimicrobial/anti-inflammatory solutions. The frequency of diagnosis was significantly higher in breeds with pendulous ears, but was not affected by ear hairiness. There were no seasonal patterns in the frequency of diagnosis. In clinical examination of 60 dogs with otitis externa, bacteria were evident in 47% of infections. Of 10 genera cultured, Staphylococcus spp. and diptheroids were most common. In this study, analysis of clinical records provided insights into the local prevalence of otitis externa and the efficacy of treatment in routine clinical situations.

Étude épidémiologique des chiens atteints d'une otite externe au cap Breton, en Nouvelle-Écosse. De mai 2008 à décembre 2013, 320 cas d'otite externe ont été diagnostiqués parmi 2012 chiens subissant des examens de routine à la Celtic Creatures Veterinary Clinic, pour une fréquence de diagnostic de 15,9 % (IC de 95 % : de 14,3 % à 17,6 %). Vingt-quatre pour cent des chiens ont manifesté une ou plusieurs récurrences malgré le traitement initial avec des solutions topiques antimicrobiennes/anti-inflammatoires. La fréquence du diagnostic était significativement supérieure chez les races avec des oreilles pendantes, mais elle n'était pas affectée par la présence de poils dans l'oreille. Il n'y avait aucun profil saisonnier dans la fréquence du diagnostic. Dans un examen clinique de 60 chiens avec une otite externe, les bactéries étaient évidentes dans 47 % des infections. Parmi les 10 genres pour lesquels une culture a été réalisée, Staphylococcus spp. et les dipthéroïdes étaient les plus fréquents. Dans cette étude, l'analyse des dossiers cliniques a fourni des renseignements sur la prévalence locale de l'otite externe et l'efficacité du traitement des situations cliniques de routine.(Traduit par Isabelle Vallières).

A review of the Polystira clade--the Neotropic's largest marine gastropod radiation (Neogastropoda: Conoidea: Turridae sensu stricto).

The Polystira clade (here comprising Polystira and Pleuroliria) is a poorly known but hyper-diverse clade within the neogastropod family Turridae (sensu stricto). It has extensively radiated within the tropics and subtropics of the Americas, to which it is endemic. In this paper we present a synthetic overview of existing information on this radiation together with new information on estimated species diversity, systematic relationships, a species-level molecular phylogenetic analysis and preliminary macroecological and diversification analyses, to serve as a platform for further study. We currently estimate that about 300 species (122 extant) are known from its 36 million year history but this number will undoubtedly increase as we extend our studies. We discuss the relationships of Polystira to other Neotropical Turridae (s.s.) and examine the taxonomy and systematics of the geologically oldest described members of the clade. To aid taxonomic description of shells we introduce a new notation for homologous major spiral cords. Focusing on key publications, we discuss in detail the changing historical understanding of the taxonomy of the clade and the relationships of its component genus-level taxa: Polystira Woodring, 1928, Pleuroliria de Gregorio, 1890, Josephina Gardner, 1945 and Oxytropa Glibert, 1955. We designate a neotype for Pleurotoma (Pleuroliria) supramirifica de Gregorio, 1890, to stabilize our understanding of this, the type species of Pleuroliria. Application of the name Oxytropa is restricted to the type species. The genus Polystira is conchologically re-described and for the first time we synthesize available information on the anatomy, feeding and toxinology, reproduction and life history, larval modes and life habits, and geographic and bathymetric ranges of its species. We give an updated list of the 19 formally described living species and present the pitfalls of the currently poor species-level taxonomy of Polystira using case examples. We present a molecular phylogenetic analysis of 22 extant species using three mitochondrial gene fragments (COI, 12S rRNA and 16S rRNA). This reveals undescribed species and indicates that Recent genetic clades ('biospecies') are consistent with finely divided conchological 'morphospecies'. Historically, there has been a slow realisation of the high species diversity of the Polystira clade and we consider that this may be due to inadequate precision of morphological description of shells and a lack of clear homology statements. We suggest how these both might be improved. Finally, using a data compilation based on museum specimens we examine species range-size distributions and species abundance distributions for 85 of the 112 extant western Atlantic species that we have delimited to date. Our results indicate that the majority of species are rare and have short geographic ranges; only a few are wide-ranging and abundant. This has important implications for surveys of biodiversity.

Sessile snails, dynamic genomes: gene rearrangements within the mitochondrial genome of a family of caenogastropod molluscs.

BACKGROUND: Widespread sampling of vertebrates, which comprise the majority of published animal mitochondrial genomes, has led to the view that mitochondrial gene rearrangements are relatively rare, and that gene orders are typically stable across major taxonomic groups. In contrast, more limited sampling within the Phylum Mollusca has revealed an unusually high number of gene order arrangements. Here we provide evidence that the lability of the molluscan mitochondrial genome extends to the family level by describing extensive gene order changes that have occurred within the Vermetidae, a family of sessile marine gastropods that radiated from a basal caenogastropod stock during the Cenozoic Era. RESULTS: Major mitochondrial gene rearrangements have occurred within this family at a scale unexpected for such an evolutionarily young group and unprecedented for any caenogastropod examined to date. We determined the complete mitochondrial genomes of four species (Dendropoma maximum, D. gregarium, Eualetes tulipa, and Thylacodes squamigerus) and the partial mitochondrial genomes of two others (Vermetus erectus and Thylaeodus sp.). Each of the six vermetid gastropods assayed possessed a unique gene order. In addition to the typical mitochondrial genome complement of 37 genes, additional tRNA genes were evident in D. gregarium (trnK) and Thylacodes squamigerus (trnV, trnLUUR). Three pseudogenes and additional tRNAs found within the genome of Thylacodes squamigerus provide evidence of a past duplication event in this taxon. Likewise, high sequence similarities between isoaccepting leucine tRNAs in Thylacodes, Eualetes, and Thylaeodus suggest that tRNA remolding has been rife within this family. While vermetids exhibit gene arrangements diagnostic of this family, they also share arrangements with littorinimorph caenogastropods, with which they have been linked based on sperm morphology and primary sequence-based phylogenies. CONCLUSIONS: We have uncovered major changes in gene order within a family of caenogastropod molluscs that are indicative of a highly dynamic mitochondrial genome. Studies of mitochondrial genomes at such low taxonomic levels should help to illuminate the dynamics of gene order change, since the telltale vestiges of gene duplication, translocation, and remolding have not yet been erased entirely. Likewise, gene order characters may improve phylogenetic hypotheses at finer taxonomic levels than once anticipated and aid in investigating the conditions under which sequence-based phylogenies lack resolution or prove misleading.

The identity, distribution, and impacts of non-native apple snails in the continental United States.

BACKGROUND: Since the mid 1990s populations of non-native apple snails (Ampullariidae) have been discovered with increasing frequency in the continental United States. Given the dramatic effects that introduced apple snails have had on both natural habitats and agricultural areas in Southeast Asia, their introduction to the mainland U.S. is cause for concern. We combine phylogenetic analyses of mtDNA sequences with examination of introduced populations and museum collections to clarify the identities, introduced distributions, geographical origins, and introduction histories of apple snails. RESULTS: Based on sampling to date, we conclude there are five species of non-native apple snails in the continental U.S. Most significantly, we recognize three species within what has been called the channeled apple snail: Pomacea canaliculata (California and Arizona), Pomacea insularum, (Florida, Texas, and Georgia) and Pomacea haustrum (Florida). The first established populations of P. haustrum were discovered in the late 1970s in Palm Beach County Florida, and have not spread appreciably in 30 years. In contrast, populations of P. insularum were established in Texas by 1989, in Florida by the mid to late 1990s, and in Georgia by 2005, and this species continues to spread rapidly. Most introduced P. insularum haplotypes are a close match to haplotypes from the Río Uruguay near Buenos Aires, indicating cold tolerance, with the potential to spread from Florida, Georgia, and Texas through Louisiana, Alabama, Mississippi, and South Carolina. Pomacea canaliculata populations were first discovered in California in 1997. Haplotypes of introduced P. canaliculata match native-range haplotypes from near Buenos Aires, Argentina, also indicating cold tolerance and the potential to establish farther north. CONCLUSION: The term "channeled apple snail" is descriptive of a morphology found in many apple snail species. It does not identify a single species or a monophyletic group. Clarifying species identifications permits a more accurate assessment of introduction histories and distributions, and provides a very different picture of the tempo and pattern of invasions than was inferred when the three species with channeled sutures were considered one. Matching introduced and native-range haplotypes suggests the potential for range expansion, with implications for native aquatic ecosystems and species, agriculture, and human health.

Changing identities: tRNA duplication and remolding within animal mitochondrial genomes.

Although the majority of metazoan mitochondrial genomes (mtDNAs) contain the same 37 genes, including 22 encoding transfer RNAs (tRNAs), the recognition of orthologs is not always straightforward. Here we demonstrate that inferring tRNA orthologs among taxa by using anticodon triplets and deduced secondary structure can be misleading: through a process of tRNA duplication and mutation in the anticodon triplet, remolded leucine (LUUR) tRNA genes have repeatedly taken over the role of isoaccepting LCUN leucine tRNAs within metazoan mtDNA. In the present work, data from within the gastropods and a broad survey of metazoan mtDNA suggest that tRNA leucine duplication and remolding events have occurred independently at least seven times within three major animal lineages. In all cases where the mechanism of gene remolding can be inferred with confidence, the direction is the same: from LUUR to LCUN. Gene remolding and its apparent asymmetry have significant implications for the use of mitochondrial tRNA gene orders as phylogenetic markers. Remolding complicates the identification of orthologs and can result in convergence in gene order. Careful sequence-based analysis of tRNAs can help to recognize this homoplasy, improving gene-order-based phylogenetic hypotheses and underscoring the importance of careful homology assessment. tRNA remolding also provides an additional mechanism by which gene order changes can occur within mtDNA: through the changing identity of tRNA genes themselves. Recognition of these remolding events can lead to new interpretations of gene order changes, as well as the discovery of phylogenetically relevant gene dynamics that are hidden at the level of gene order alone.

ENCAPSULATION OF EGGS BY MARINE GASTROPODS: EFFECT OF VARIATION IN CAPSULE FORM ON THE VULNERABILITY OF EMBRYOS TO PREDATION.

Representatives of many plant and animal taxa enclose their embryos within some form of protective structure. Inter- and intraspecific differences in the morphology of these egg coverings may have profound effects on the development and survival of encapsulated embryos, yet in many taxa little is known about the causes or potential consequences of this variation. Comparisons of capsule morphology among populations of the rocky shore gastropod, Nucella emarginata, revealed significant variation in the thickness of capsule walls, the only barrier separating developing embryos from the external environment. Laboratory experiments demonstrated that thicker-walled capsules were more resistant to predation by a co-occurring isopod, Gnorimosphaeroma oregonense, than were thinner-walled capsules. Control experiments confirmed that these differences in vulnerability were not caused by differences in the palatability of the capsule wall or attractiveness of the capsule contents. The actual mechanism by which thick-walled capsules differentially protect developing embryos remains unclear, although decreased vulnerability of thick-walled capsules to these isopods may simply result from increased handling time by predators. Subtle differences in capsule morphology thus appear to have substantial effects on the survival of encapsulated embryos. Hence, predators may have played an important role in selecting for the production of thick-walled capsules among populations of N. emarginata.