A Phylogenomic Backbone for Gastropod Molluscs.

Gastropods have survived several mass extinctions during their evolutionary history resulting in extraordinary diversity in morphology, ecology, and developmental modes, which complicate the reconstruction of a robust phylogeny. Currently, gastropods are divided into six subclasses: Caenogastropoda, Heterobranchia, Neomphaliones, Neritimorpha, Patellogastropoda, and Vetigastropoda. Phylogenetic relationships among these taxa historically lack consensus, despite numerous efforts using morphological and molecular information. We generated sequence data for transcriptomes derived from 12 taxa belonging to clades with little or no prior representation in previous studies in order to infer the deeper cladogenetic events within Gastropoda and, for the first time, infer the position of the deep-sea Neomphaliones using a phylogenomic approach. We explored the impact of missing data, homoplasy, and compositional heterogeneity on the inferred phylogenetic hypotheses. We recovered a highly supported backbone for gastropod relationships that is congruent with morphological and mitogenomic evidence, in which Patellogastropoda, true limpets, are the sister lineage to all other gastropods (Orthogastropoda) which are divided into two main clades 1) Vetigastropoda $s.l.$ (including Pleurotomariida $+$ Neomphaliones) and 2) Neritimorpha $+$ (Caenogastropoda $+$ Heterobranchia). As such, our results support the recognition of five subclasses (or infraclasses) in Gastropoda: Patellogastropoda, Vetigastropoda, Neritimorpha, Caenogastropoda, and Heterobranchia. [Compositional heterogeneity; fast-evolving; long-branch attraction; missing data; Mollusca; phylogenetics; systematic error.].

A biting commentary: Integrating tooth characters with molecular data doubles known species diversity in a lineage of sea slugs that consume "killer algae".

Predicting biotic resistance to highly invasive strains of "killer algae" (Caulerpa spp.) requires understanding the diversity and feeding preferences of native consumers, including sea slugs in family Oxynoidae. Past studies reported low algal host specificity for Oxynoe (6 spp.) and Lobiger (4 spp.), but these taxonomically challenging slugs may represent species complexes of unrecognized specialists that prefer different Caulerpa spp. Here, we assess global diversity of these genera by integrating gene sequences with morphological data from microscopic teeth and internal shells, the only hard parts in these soft-bodied invertebrates. Four delimitation methods applied to datasets comprising mtDNA and/or nuclear alleles yielded up to 16 species hypotheses for samples comprising five nominal taxa, including five highly divergent species in Lobiger and five in Oxynoe. Depending on the analysis, a further four to six species were recovered in the O. antillarum-viridis complex, a clade in which mitochondrial divergence was low and nuclear alleles were shared among lineages. Bayesian species delimitation using only morphological data supported most candidate species, however, and integrative analyses combining morphological and genetic data fully supported all complex members. Collectively, our findings double the recognized biodiversity in Oxynoidae, and illustrate the value of including data from traits that mediate fast-evolving ecological interactions during species delimitation. Preference for Caulerpa spp. and radular tooth characteristics covaried among newly delimited species, highlighting an unappreciated degree of host specialization and coevolution in these taxa that may help predict their role in containing outbreaks of invasive algae.

Deep molecular divergence and exceptional morphological stasis in dwarf cannibal snails Nata sensu lato Watson, 1934 (Rhytididae) of southern Africa.

The genus Nata Watson, 1934 is a southern African endemic belonging to the Gondwanan family of carnivorous snails, Rhytididae. We present a molecular phylogeny of the genus based on two mitochondrial (16S and COI) and two nuclear genes (ITS2 and 28S RNA), and complement this with an appraisal of morphological characters relating to both the shell and soft parts. We identify four reciprocally monophyletic lineages for which valid names are already available, plus two undescribed species restricted to the Albany Thicket Biome. We show that Nata sensu lato may not be monophyletic. Rather there exist two deep lineages within Nata s.l., one lineage potentially sister to a clade dominated by the Australian and New Zealand radiation, and the other occupying a basal position within Rhytididae. Accordingly we recommend a revision recognising two genera, namely Nata s.s. and Natella respectively. Despite deep molecular divergences within Nata s.s., phenotypic evolution has been remarkably conserved, and contrasts greatly with that exhibited across other major lineages within the Rhytididae.

The rhipidoglossan radula: Radular morphology and formation in Nerita litterata Gmelin, 1791 (Neritimorpha, Neritidae).

The rhipidoglossan radula, consisting of numerous teeth in each transverse row, is characteristic of phylogenetically distant groups of gastropods, including Vetigastropoda, Neritimorpha and 'lower' Heterobranchia. Previous studies have revealed the main patterns in the formation of the rhipidoglossan radula of vetigastropods, the main feature of which is the division of the formation zone into two horns, where marginal teeth are formed by a multilayered epithelium (odontoblasts). This work is devoted to the study of the formation of the rhipidoglossan radula of Nerita litterata using light and electron microscopy. The data obtained show that, despite the different external morphology of the radular sac of neritids and vetigastropods, the radular sac of N. litterata, like that of vetigastropods, is divided into two parts, in which the marginal teeth are similarly formed by odontoblasts located in more than one layer. It seems probable that this complex, three-dimensional structure of the formation zone is associated with a broad radula with numerous elongate marginal teeth and could be characteristic of other gastropods with this type of radula. Additional supporting rods located along the odontoblasts and consisting of vacuolated cells were first discovered in Nerita.

The rhipidoglossan radula: Formation and morphology of the radula in Puncturella noachina (Linnaeus, 1771) (Fissurellidae, Vetigastropoda).

The rhipidoglossan radula, which is characterized by presence of a central tooth, several lateral teeth, and numerous (more than 10) long marginal teeth in each transverse row, is found in three different subclasses, that is, Vetigastropoda, Neritomorpha and "lower" Heterobranchia. Details of radula formation and its ultrastructure have not been studied in any species with a rhipidoglossan radula. For the first time, we present such data for one vetigastropod species, Puncturella noachina. The radula itself and the radula formation zone were studied using light and electron microscopy (scanning and transmission), as well as confocal laser scanning microscopy. We identify the major features of Vetigastropoda rhipidoglossan radula formation, that is: the posterior bifurcation of the radula formation zone, creating paired horns into which the zones of formation of the marginal teeth extend; the supporting structure in the radula formation zone extends ventrally to strengthen this division; the odontoblasts of the marginal teeth form a multi-layered epithelium; membranoblasts do not differ from odontoblasts in ultrastructure; in some membranoblasts and cells of the sub- and supraradular epithelium basal bodies were found in the apical regions of the cells.

The rhipidoglossan radula: Formation and development in Margarites helicinus Phipps, 1774 (Trochoidea, Vetigastropoda).

The gastropod radula exhibits exceptional morphological variability. Despite this enormous diversity, the main patterns of synthesis of the teeth and radula membrane, characteristic of different groups can be identified. The rhipidoglossan radula contains numerous teeth in each transverse row and was found in phylogenetically distant groups of Gastropoda. Studying radula formation through stages of ontogeny in species with this type of radula is important in determining the main patterns of radula formation in gastropods. We studied the formation of the radula during development of one species of trochid vetigastropod, Margarites helicinus, using light and electron microscopy as well as confocal laser scanning microscopy. The adult radula is formed in the blind end of the radular sac, which bifurcates into two horns in vetigastropods. The numerous marginal teeth are synthesized in these horns while the central and lateral teeth form in the region where the horns fuse. This morphology of the formation zone appears to be a common pattern for all vetigastropods. The juvenile radula of M. helicinus consists of seven teeth per transverse row and its formation in the radular sac differs significantly from that in the adult. In the juvenile, the formation zone of the radular sac is not split into two horns, and the teeth and radular membrane are synthesized by relatively few, uniform cells. This organization of the larval radular sac is thought to represent a widely occurring larval pattern potentially present in all groups of gastropods. It is associated with early formation of all organs by few cells rather than representing a phylogenetic trait.

Land snail biogeography and endemism in south-eastern Africa: Implications for the Maputaland-Pondoland-Albany biodiversity hotspot.

Invertebrates in general have long been underrepresented in studies on biodiversity, biogeography and conservation. Boundaries of biodiversity hotspots are often delimited intuitively based on floristic endemism and have seldom been empirically tested using actual species distributions, and especially invertebrates. Here we analyse the zoogeography of terrestrial malacofauna from south-eastern Africa (SEA), proposing the first mollusc-based numerical regionalisation for the area. We also discuss patterns and centres of land snail endemism, thence assessing the importance and the delimitation of the Maputaland-Pondoland-Albany (MPA) biodiversity hotspot for their conservation. An incidence matrix compiled for relatively well-collected lineages of land snails and slugs (73 taxa in twelve genera) in 40 a priori operational geographic units was subjected to (a) phenetic agglomerative hierarchical clustering using unweighted pair-group method with arithmetic means (UPGMA), (b) parsimony analysis of endemicity (PAE) and biotic element analysis (BEA). Fulfilling the primary objective of our study, the UPGMA dendrogram provided a hierarchical regionalisation and identified five centres of molluscan endemism for SEA, while the PAE confirmed six areas of endemism, also supported by the BEA. The regionalisation recovers a zoogeographic province similar to the MPA hotspot, but with a conspicuous westward extension into Knysna (towards the Cape). The MPA province, centres and areas of endemism, biotic elements as well as the spatial patterns of species richness and endemism, support the MPA hotspot, but suggest further extensions resulting in a greater MPA region of land snail endemism (also with a northward extension into sky islands-Soutpansberg and Wolkberg), similar to that noted for vertebrates. The greater MPA region provides a more robustly defined region of conservation concern, with centres of endemism serving as local conservation priorities.

Preparedness of Australasian emergency departments for point-of-care ultrasound in the COVID-19 pandemic.

INTRODUCTION: Point-of-care ultrasound (POCUS) has been brought to the limelight again, with a surge in lung ultrasound in suspected COVID-19 patients. This is due to POCUS superiority over chest X-ray, equivalent efficacy to computerised tomography chest for COVID-19 diagnosis and potential minimisation of cross-infection. However, inadequate disinfection practices could make ultrasound machines a vector for disease transmission. This study, conducted during the early phase of the COVID-19 pandemic, surveyed the preparedness of Australasian Clinicians for responsible POCUS practice within the Emergency Department (ED). METHODS: An anonymous online survey conducted from 20th April to 3rd June 2020 among emergency clinicians providing POCUS within Australasian EDs investigated preparedness to provide effective POCUS while minimising cross-infection. RESULTS: The survey received 171 responses and 116 being eligible for analysis. Most respondents (n = 96, 98%) had a separate 'hot zone' with a dedicated US device (n = 75, 77%), but lacked COVID-19-specific standard-operating procedures (n = 51, 52%) or a designated safety and compliance officer (n = 36, 37%). Most clinicians (n = 86, 88%) were willing to perform ultrasound in highly infectious patients, despite poor formal training (n = 66, 67%) or COVID-19-specific lung protocols (n = 59, 60%). Most (n = 92, 93%) had access to appropriate low-level disinfectant wipes but varied significantly in disinfection practice due to a lack of timely, formal or unified guidelines. CONCLUSION: Australasian EDs significantly lacked investment in education, training and protocols to conduct safe POCUS in the COVID-19 pandemic. A framework with evidence-based, logistically feasible protocols supporting safe emergency POCUS is required to deal with similar future infectious outbreaks.

Radula formation in two species of Conoidea (Gastropoda).

The radula is the basic feeding structure in gastropod molluscs and exhibits great morphological diversity that reflects the exceptional anatomical and ecological diversity occurring in these animals. This uniquely molluscan structure is formed in the blind end of the radular sac by specialized cells (membranoblasts and odontoblasts). Secretion type, and the number and shape of the odontoblasts that form each tooth characterize the mode of radula formation. These characteristics vary in different groups of gastropods. Elucidation of this diversity is key to identifying the main patterns of radula formation in Gastropoda. Of particular interest would be a phylogenetically closely related group that is characterized by high variability of the radula. One such group is the large monophyletic superfamily Conoidea, the radula of which is highly variable and may consist of the radular membrane with five teeth per row, or the radular membrane with only two or three teeth per row, or even just two harpoon-like teeth per row without a radular membrane. We studied the radulae of two species of Conoidea (Clavus maestratii Kilburn, Fedosov & Kantor, 2014 [Drilliidae] and, Lophiotoma acuta (Perry, 1811) [Turridae]) using light and electron microscopy. Based on these data and previous studies, we identify the general patterns of the radula formation for all Conoidea: the dorsolateral position of two groups of odontoblasts, uniform size, and shape of odontoblasts, folding of the radula in the radular sac regardless of the radula configuration. The morphology of the subradular epithelium is most likely adaptive to the radula type.

A phylogeny of the cannibal snails of southern Africa, genus Natalina sensu lato (Pulmonata: Rhytididae): assessing concordance between morphology and molecular data.

The genus Natalina Pilsbry, 1893 is a southern African endemic belonging to the Gondwanan family of carnivorous snails, Rhytididae. We present a well-resolved molecular phylogeny of the genus based on the mitochondrial 16S and COI genes and the nuclear ITS2 gene, and assess this in light of Watson's [Watson, H., 1934. Natalina and other South African snails. Proc. Malacol. Soc. Lond. 21, 150-193] supra-specific classification via a re-examination of 23 morphological characters including features of the shell, radula, external anatomy and distal reproductive tract. Ancestral reconstruction and character mapping based on the MK(1) model reveals broad concordance between morphology and the molecular phylogeny at the supra-specific level. Given this concordance and exceptionally deep divergences in the molecular data, we recommend the elevation of the subgenera Natalina s.s., Afrorhytida, and Capitina to generic status. At the species level, we identify several species complexes for which additional fine scale morphological and molecular appraisal is needed to qualify on the one hand incipient speciation with notable differentiation in shell form and body pigmentation, and on the other, phylogenetically deep yet morphologically cryptic diversity.

Revision of the dwarf cannibal snails (Nata s.l.) of southern Africa-Nata s.s. and Natella (Mollusca: Gastropoda: Rhytididae), with description of three new species.

This paper represents the second part of our revisionary studies on the rhytidid fauna of southern Africa. The species discussed belong to the taxon Nata s.l. in which we recognise two genus-level lineages, Nata s.s. and Natella with six (three new) and one species respectively. We update the species-level taxonomy extensively in the light of new molecular and morphological data, and provide a comprehensive revision of all species, including keys. Detailed comparative morphological observations are provided for the distal reproductive tract, pulmonary cavity, mantle edge, radula and suprapedal gland. In addition, we present a summary of biological and ecological data including information on habitat preferences, feeding, prey items and mating behaviour. Although the two genera are well circumscribed in terms of both internal anatomy and molecular data, shell morphology is highly conserved, and species discrimination using shell characters alone is difficult. We have discovered three undescribed species within Nata and there is evidence that further research of a phylogeographic nature may uncover additional cryptic diversity. The geographic distributions of the species are discussed in relation to regional vegetation patterns and, as with the larger cannibal snail radiation, the Albany Thicket Biome emerges as a focus of endemism. Observations on the conservation status of all species are provided.New species: Nata aequiplicata sp. nov., Nata erugata sp. nov. and Nata watsoni sp. nov. Revised status: Natalina caffrula Melvill & Ponsonby, 1898 is transferred to the genus Nata and thought to be a synonym of Nata dumeticola (Benson, 1851).

Prestonellinae-validation of the name as a new subfamily of Bothriembryontidae (Mollusca, Gastropoda, Orthalicoidea).

The affinities of the enigmatic South African land snail genus Prestonella Connolly, 1929 were discussed by Herbert (2007) and Herbert & Mitchell (2009), who showed, on the basis of morphological and molecular data, that the genus is referable to the superfamily Orthalicoidea. Currently, the three described species of Prestonella are the only known African representatives of this diverse superfamily. Earlier, van Bruggen (1978) had recognized that these species formed a distinct group and had placed them in the (new) family Prestonellidae. However, as noted by Bouchet & Rocroi (2005: 140), no diagnosis was provided by van Bruggen; the name Prestonellidae thus does not meet the requirements of ICZN Art. 13.1, and is not an available name. In this paper we will redress this issue, also taking into account more recent research which has shed light on the systematic position of this genus within the Orthalicoidea.

An annotated catalogue and bibliography of the taxonomy, synonymy and distribution of the Recent Vetigastropoda of South Africa (Mollusca).

A complete inventory of the known Recent vetigastropod fauna of South Africa is provided. Bibliographic citations to works discussing the taxonomy, synonymy and distribution of the species in a southern African or south-western Indian Ocean context are provided. Additional explanatory notes are given where pertinent. New genus records for South Africa: Acremodontina B.A. Marshall, 1995; Choristella Bush, 1879; Cocculinella Thiele, 1909; Conjectura Finlay, 1926; Crosseola Iredale, 1924; Falsimargarita Powell, 1951; Lepetella Verrill, 1880; Profundisepta McLean & Geiger, 1998; Stomatella Lamarck, 1816; Stomatia Helbling, 1779; Stomatolina Iredale, 1937; Synaptocochlea Pilsbry, 1890; Tibatrochus Nomura, 1940; Visayaseguenzia Poppe, Tagaro & Dekker, 2006; Zetela Finlay, 1926. New species records for South Africa: Acremodontina aff. carinata Powell, 1940; Anatoma finlayi (Powell, 1937); Anatoma munieri (P. Fischer, 1862); Calliotropis acherontis B.A. Marshall, 1979; Calliotropis bucina Vilvens, 2006; Cocculinella minutissima (E.A. Smith, 1904); Diodora ruppellii (G.B. Sowerby (I), 1835); Emarginula costulata Deshayes, 1863; Emarginula decorata Deshayes, 1863; Jujubinus hubrechti Poppe, Tagaro & Dekker, 2006; Lepetella sp.; Seguenzia orientalis Thiele, 1925; Stomatella auricula Lamarck, 1816; Stomatia phymotis Helbling, 1779; Stomatolina angulata (A. Adams, 1850); Stomatolina cf. calliostoma (A. Adams, 1850); Stomatolina aff. danblumi Singer & Mienis, 1999; Stomatolina cf. rubra (Lamarck, 1822); Stomatolina sp.; Synaptocochlea concinna (Gould, 1845); Tectus mauritianus (Gmelin, 1791); Tibatrochus cf. incertus (Schepman, 1908); Turbo imperialis Gmelin, 1791; Turbo tursicus Reeve, 1848; Visayaseguenzia compsa (Melvill, 1904).New species: Spectamen martensi, replacement name for Spectamen semisculptum sensu Herbert (1987) (non Martens, 1904). New name: Oxystele antoni is proposed as a new name for Trochus (Turbo) variegatus (non Gmelin, 1791 =Heliacus) Anton, 1838. Revised taxonomy: Cyclostrema semisculptum Martens, 1904 is an earlier name for Solariella intermissa Thiele, 1925, and is referable to the genus Zetela Finlay, 1926; Margarita bicarinata A. Adams & Reeve, 1850 is considered to be a senior synonym of Solariella undata G.B. Sowerby (II), 1870, and is referable to the genus Ilanga Herbert, 1987. Validation of the name Trochus tigrinus Chemnitz, 1781 is credited to Dillwyn (1817) rather than Anton (1838). New synonyms: Clanculus exquisita Turton, 1932 =Calliostoma africanum Bartsch, 1915; Cyclostrema alfredensis Bartsch, 1915 =Parviturbo alfredensis (Bartsch, 1915); Cynisca gloriosa Bartsch, 1915 =Cinysca spuria (Gould, 1861); Herbertina hayesi Herbert, 1995 =Bruceina chenoderma (Barnard, 1963); Ilanga millardi Herbert, 1987 =Ilanga humillima (Thiele, 1925); Leptothyra africana Bartsch, 1915 =Cinysca spuria (Gould, 1861); Leptothyra albocincta Turton, 1932 =Tricolia striolata (Turton, 1932); Solariella undata G.B. Sowerby (II), 1870, S. gratiosa Thiele, 1925 and S. valdiviae Thiele, 1925 =Ilanga bicarinata bicarinata (A. Adams & Reeve, 1850); Solariella chuni Thiele, 1925, S. intermissa Thiele, 1925, S. gilchristi Barnard, 1963 and S. macleari Barnard, 1963 =Zetela semisculpta (Martens, 1904); Turbo (Collonia) armillatus G.B. Sowerby (III), 1886 =Cinysca spuria (Gould, 1861). New combinations: Basilissa (Ancistrobasis) compsa Melvill, 1904 is transferred to Visayaseguenzia; Calcar rhysopoma Barnard, 1964 is transferred to Bothropoma; Calliostoma glaucophaos Barnard, 1963 is transferred to Falsimargarita; Calliotropis chenoderma Barnard, 1963 is transferred to Bruceina; Collonia bicarinata Martens, 1902 is transferred to Cinysca; Crossea agulhasensis Thiele, 1925 is transferred to Conjectura; Cyclostrema semisculptum Martens, 1904 is transferred to Zetela; Cyclostremella farica Bartsch, 1915 is transferred to Dikoleps; Cynisca africana Bartsch, 1915 is transferred to Homalopoma; Leptogyra africana: Bartsch, 1915 is transferred to Cirsonella; Leptothyra agulhasensis Thiele, 1925 is transferred to Homalopoma; Leptothyra alfredensis Bartsch, 1915 is transferred to Parviturbo; Leptothyra sola Barnard, 1963 is transferred to a Parviturbo; Liotia (Cynisca) semiclausa Thiele, 1925 is transferred to Cinysca; Monilea spuria Gould, 1861 is transferred to Cinysca; Monodonta gibbula Thiele, 1925 is transferred to Cantrainea; Puncturella voraginosa Herbert & Kilburn, 1986 is transferred to Profundisepta; Solariella fuscomaculata G.B. Sowerby (III), 1892 is transferred to Skenea; Solariella turbynei Barnard, 1963 is transferred to Zetela; Turbo boswellae Barnard, 1969 is transferred to Cantrainea; Turbo foveolatus Barnard, 1963 is transferred to Crosseola; Turbo ponsonbyi G.B. Sowerby (III), 1897 is transferred to Bothropoma; Vitrinella agulhasensis Thiele, 1925 is transferred to Parviturbo; Vitrinella (Docomphala) arifca Bartsch, 1915 is transferred to Lodderena; Vitrinella inclinans Barnard, 1963 is transferred to Skenea.

Prolonged venovenous extracorporeal membrane oxygenation without anticoagulation: a case of Goodpasture syndrome-related pulmonary haemorrhage.

We present a 16-year-old male with severe acute respiratory and renal failure as a result of Goodpasture syndrome, requiring venovenous extracorporeal membrane oxygenation (VV-ECMO) for pulmonary haemorrhage. The patient received no systemic anticoagulation for 25 of 26 ECMO days (20 days consecutively) and suffered no coagulation-related adverse events. The patient had a subtherapeutic anticoagulation profile according to recommended ECMO guidelines during most of this time. The patient made a full recovery without respiratory compromise, ECMO circuit failure, thrombotic events or the need for ongoing haemodialysis.

New genus Arxellia with new species of Solariellidae (Gastropoda: Trochoidea) from New Caledonia, Papua New Guinea, Philippines, Western Australia, Vanuatu and Tonga.

A new genus, Arxellia, is described in the family Solariellidae. Nine species are referred to this taxon, eight of which are new and are described in this paper (Arxellia trochos n. sp., Arxellia boucheti n. sp., Arxellia herosae n. sp., Arxellia helicoides n. sp., Arxellia tracheia n. sp., Arxellia thaumasta n. sp., Arxellia maestratii n. sp. and Arxellia erythrea n. sp.). The previously described species Bathymophila tenorioi Poppe, Tagaro & Dekker, 2006 is reassigned to Arxellia.