Unveiling underestimated species diversity within the Central American Coralsnake, a medically important complex of venomous taxa

Coralsnakes of the genus Micrurus are a diverse group of venomous snakes ranging from the southern United States to southern South America. Much uncertainty remains over the genus diversity, and understanding Micrurus systematics is of medical importance. In particular, the widespread Micrurus nigrocinctus spans from Mexico throughout Central America and into Colombia, with a number of described subspecies. This study provides new insights into the phylogenetic relationships within M. nigrocinctus by examining sequence data from a broad sampling of specimens from Mexico, Guatemala, Honduras, Nicaragua, Costa Rica, and Panama. The recovered phylogenetic relationships suggest that M. nigrocinctus is a species complex originating in the Pliocene and composed of at least three distinct species-level lineages. In addition, recovery of highly divergent clades supports the elevation of some currently recognized subspecies to the full species rank while others may require synonymization.National Science Foundation (grant no. DEB-0416160 to ENS),Instituto Bioclon (grant to ENS)UTA-Biology startup funds (to ENS)Portuguese Foundation for Science and Technology (FCT, fellowship number SFRH/ BPD/109148/2015)Maria Zambrano grant from the Spanish Ministry of Science and Education and Next Generation EU.Brazilian National Council of Technological and Scientific Development (CNPq) (314287/2020-5)European Union’s Horizon 2020 Research and Innovation Programme under the Grant Agreement Number 85725

Treerunners, cryptic lizards of the Plica plica group (Squamata, Sauria, Tropiduridae) of northern South America

The arboreal, Neotropical lizard Plica plica (Linnaeus, 1758) has been long considered a widespread species with a distribution east of the Andes. A preliminary examination of 101 specimens from about 28 locations mostly north of the Amazon suggests that Plica plica is a cryptic species complex with taxa that can be distinguished on the basis of the number of scale rows at mid-body; the arrangement, shape and ornamentation of scales on the snout; the number of lamellae on the fourth toe; the number of subocular plates; as well as other commonly used external morphological traits. The allopatric species discussed here are concordant with northern South American geography. Plica plica (Linnaeus,1758) is associated with the Guiana Shield (Suriname, Guyana and Venezuela). A second species, P. caribeana sp. n. is associated with the Caribbean Coastal Range of Venezuela including Trinidad and Tobago. A third, distinctive species, P. rayi sp. n. is associated with the middle Orinoco at the eastern edge of the Guiana Shield. Two other species, P. kathleenae sp. n. and P. medemi sp. n., each based upon a single specimen, one from the Sierra Acarai Mountains of Guyana, and the other from southern Meta, Colombia are described. In addition to morphological analyses, we sequenced 12S and 16S rDNA gene fragments from one Plica plica from Trinidad to assess its relationship and taxonomy to other mainland Plica cf plica. The results suggest Plica caribeana sp. n. likely diverged prior to the separation of Trinidad from northern Venezuela. Isolation in the Caribbean Coastal Range during its rapid uplift in the late Miocene, combined with a marine incursion into northern Venezuela may have contributed to their genetic divergence from other populations. © John C. Murphy, Michael Jowers.Peer Reviewe

Treerunners, cryptic lizards of the Plica plica group (Squamata, Sauria, Tropiduridae) of northern South America

The arboreal, Neotropical lizard Plica plica (Linnaeus, 1758) has been long considered a widespread species with a distribution east of the Andes. A preliminary examination of 101 specimens from about 28 locations mostly north of the Amazon suggests that Plica plica is a cryptic species complex with taxa that can be distinguished on the basis of the number of scale rows at mid-body; the arrangement, shape and ornamentation of scales on the snout; the number of lamellae on the fourth toe; the number of subocular plates; as well as other commonly used external morphological traits. The allopatric species discussed here are concordant with northern South American geography. Plica plica (Linnaeus,1758) is associated with the Guiana Shield (Suriname, Guyana and Venezuela). A second species, P. caribeana sp. n. is associated with the Caribbean Coastal Range of Venezuela including Trinidad and Tobago. A third, distinctive species, P. rayi sp. n. is associated with the middle Orinoco at the eastern edge of the Guiana Shield. Two other species, P. kathleenae sp. n. and P. medemi sp. n., each based upon a single specimen, one from the Sierra Acarai Mountains of Guyana, and the other from southern Meta, Colombia are described. In addition to morphological analyses, we sequenced 12S and 16S rDNA gene fragments from one Plica plica from Trinidad to assess its relationship and taxonomy to other mainland Plica cf plica. The results suggest Plica caribeana sp. n. likely diverged prior to the separation of Trinidad from northern Venezuela. Isolation in the Caribbean Coastal Range during its rapid uplift in the late Miocene, combined with a marine incursion into northern Venezuela may have contributed to their genetic divergence from other populations. © John C. Murphy, Michael Jowers.Peer Reviewe

New insights on patterns of genetic admixture and phylogeographic history in Iberian high mountain populations of midwife toads

Data Availability: Newly generated sequence data are available in GenBank (https://www.ncbi.nlm.nih.gov/genbank/) under the following accession numbers: OP779379-OP779400 for ND4, OP820514-OP820517 for cyt-b, OP820518-OP820519 for β-fibint7, OP846560-OP846568 for 12S, and OP851584-OP851590 for 16S. Original sequence alignments and microsatellite genotypes are available in the Zenodo repository (https://doi.org/10.5281/zenodo.7270824).Funding: This work was supported by the European Commission projects LIFE+ LIMNOPIRINEUS (LIFE13 NAT/ES/001210, PI: M.V.), LIFE RESQUE ALPYR (LIFE20 NAT/ES/00347, PI: M.V.) and Biodiversa FISHME (BiodivRestor-280, PI: M.V.) and by the Spanish Government projects FUNBIO (RTI2018-096217-B-I00, PI: M.V.), BIOOCULT (2413/2017, PI: M.V.), (Organismo Autónomo Parques Nacionales 2399/2017; PI: J.B) and Picos de Europa National Park grants for 2015 and 2016 (PI: J.B.). F.L. had a doctoral grant funded by Fundação para a Ciência e a Tecnologia (FCT, grant number PD/BD/52598/2014). M.J.J. had an International collaboration grant (National Institute of Ecology) and a postdoctoral grant funded by FCT (grant number SFRH/BPD/109148/2015). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Multiple Quaternary glacial refugia in the Iberian Peninsula, commonly known as “refugia within refugia”, allowed diverging populations to come into contact and admix, potentially boosting substantial mito-nuclear discordances. In this study, we employ a comprehensive set of mitochondrial and nuclear markers to shed light onto the drivers of geographical differentiation in Iberian high mountain populations of the midwife toads Alytes obstetricans and A. almogavarii from the Pyrenees, Picos de Europa and Guadarrama Mountains. In the three analysed mountain regions, we detected evidence of extensive mito-nuclear discordances and/or admixture between taxa. Clustering analyses identified three major divergent lineages in the Pyrenees (corresponding to the eastern, central and central-western Pyrenees), which possibly recurrently expanded and admixed during the succession of glacialinterglacial periods that characterised the Late Pleistocene, and that currently follow a ringshaped diversification pattern. On the other hand, populations from the Picos de Europa mountains (NW Iberian Peninsula) showed a mitochondrial affinity to central-western Pyrenean populations and a nuclear affinity to populations from the central Iberian Peninsula, suggesting a likely admixed origin for Picos de Europa populations. Finally, populations from the Guadarrama Mountain Range (central Iberian Peninsula) were depleted of genetic diversity, possibly as a consequence of a recent epidemic of chytridiomycosis. This work highlights the complex evolutionary history that shaped the current genetic composition of high mountain populations, and underscores the importance of using a multilocus approach to better infer the dynamics of population divergence.Spanish Government projects FUNBIO 2413/2017, RTI2018-096217-B-I00National Institute of Ecology SFRH/BPD/109148/ 2015European Commission BiodivRestor-280, LIFE13 NAT/ES/001210, LIFE20 NAT/ES/00347Fundação para a Ciência e a Tecnologia PD/BD/52598/201

Clarifying the taxonomy of some cryptic blennies (Blenniidae) in their native and introduced range

Omobranchus punctatus is native to the Indo-Pacific region and invasive in the Atlantic region, currently being considered one of the most widely distributed blenny species. However, recent molecular studies indicated that O. punctatus is a complex of species, with three divergent mtDNA lineages identified to date, stressing the need for a taxonomic revision. In this study, we used an integrative approach, combining morphological and genetic data, to shed light on the taxonomy and distribution of O. punctatus. Moreover, we provide the first genetic records of introduced populations in Brazil and discuss the introduction pattern of this species in this region. Morphological data shows that O. punctatus consists of at least five distinct and geographically restricted species: O. punctatus sensu stricto, O. dispar, O. sewalli, O. cf. kochi, and O. cf. japonicus. Species delimitation analyses performed using the mtDNA data available confirmed that O. punctatus sensu stricto, O. dispar and O. sewalli correspond to different species that started to diverge about 2.6 Mya. Furthermore, O. sewalli was identified as the invasive species colonizing Atlantic shores. The existence of historical oceanographic barriers, such as the emergence of the Sunda Shelf in the Eastern Indian Ocean during the Pleistocene, and the biological traits of these blennies are the most likely factors responsible for their genetic differentiation and subsequent speciation.Fundacao de Amparo a Pesquisa e Desenvolvimento Cientifico do Maranhao (FAPEMA) BEPP 03654/15Portuguese Foundation for Science and Technology SFRH/BPD/109148/2015European Union's Horizon 2020 Research and Innovation Programme 85725

Osteology, natural history notes, and phylogenetic relationships of the poorly known Caribbean frog Leptodactylus nesiotus (Anura, Leptodactylidae)

The Leptodactylus melanonotus group consists of 15 species, but references to skeletal characters are available for only three species: L. leptodactyloides, L. melanonotus, and L. diedrus. Leptodactylus nesiotus is a member of the melanonotus group known only from the type locality, Bonasse swamp, on the Southwestern peninsula of Trinidad, Trinidad and Tobago. This species has been categorized as vulnerable given its restricted distribution. Herein, we report the adult osteology of L. nesiotus, the skeletal characters are compared with the available data from other Leptodactylusspecies. A phylogenetic analysis recovers a paraphyletic L. melanonotus group relative to the L. latrans group. A monophyletic “latrans-melanonotus” clade is supported by five synapomorphies. L. nesiotus is recovered as the sister species of L. validus, a relationship supported by two synapomorphies: T-shaped terminal phalanges and a dark-colored stripe on the outer surface of arm. In addition, we report on the ecology of this poorly known species.El grupo L. melanonotus del género Leptodactylus incluye 15 especies, aunque sólo existen reportes de caracteres osteológicos para tres de ellas: L. leptodactyloides, L. melanonotus y L. diedrus. Leptodactylus nesiotus es un miembro del grupo L. melanonotus que ha sido categorizado como vulnerable debido a su distribución restringida, ya que es conocido sólo de la localidad tipo: La ciénaga de Bonasse, sudoeste de la península de la isla Trinidad, Trinidad y Tobago. En este trabajo se aportan datos sobre la osteología de adultos de L. nesiotus, los cuales son comparados con lo observado en otras especies del género, y se sumaron a una matriz previamente elaborada. Esta fuente de caracteres se utilizó para realizar un análisis de parsimonia y proponer una hipótesis filogenética. El grupo L. melanonotus resultó parafilético respecto al grupo L. latrans. El clado latrans-melanonotus es apoyado por cinco sinapomorfías. Se infiere a Leptodactylus nesiotus como la especie hermana de L. validus, relación apoyada por dos sinapomorfías: falanges terminales en forma de T, y presencia de banda oscura en el borde externo del brazo. Además se reportan observaciones sobre la ecología de esta especie pobremente conocida.Fil: Ponssa, María Laura. Fundación Miguel Lillo. Dirección de Zoología. Instituto de Herpetología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - Tucumán. Unidad Ejecutora Lillo; ArgentinaFil: de Sa, Rafael O.. University of Richmond; Estados UnidosFil: Jowers, Michael J.. University of Glasgow; Reino Unid

Slow and steady saves the race: molecular and morphological analysis of three new cryptic species of Iberus land snails from the Iberian Peninsula

The Iberian Peninsula constitutes a diversity hotspot with a high number of endemisms, where the land snail genus Iberus is likely the best example. Despite this, its species diversity is still debated as it holds several cryptic species. In the present paper, we use molecular evidence (mitochondrial DNA cytochrome oxidase subunit I) to clarify the position of I. ortizi and three new cryptic species that are described herein: I. giennensis, I. axarciensis and I. antikarianus spp. nov. For this, we sampled 281 sampling points to delimitate a comprehensive geographic mapping of these species. Moreover, we carried out a comprehensive morphometric analysis based on 3205 shells. Our findings show that, morphologically, the three described species overlap in the form of their shells, their morphologies being very similar to other close species with nearby distributions (I. ortizi, I. angustatus and I. marmoratus loxanus). Still, all these species are well-defined by genetic distances, but display allopatric distributions, suggesting that they evolved by allopatric speciation as a consequence of biogeographic isolation. Hence, our findings show insights into the evolution of land snails in southeastern Spain, with implications for their conservation, given that our exhaustive sampling shows that the three species described here have very limited distribution ranges, especially I. antikarianus sp. nov. Our study, moreover, implies an integrated approach to the study of the evolution of land snails, including the sampling of the complete geographic area occupied by the genus, genetic analysis to delimit the actual species range, as well as morphometric analyses to understand the phenotypic differentiation and adaptations of the three new species.Funding for open access publishing: Universidad de Granada/CBUAPostdoctoral “María Zambrano” fellowshi

Applicability of molecular markers to determine parasitic infection origins in the animal trade: a case study from Sarcoptes mites in wildebeest

The development of non-manipulative molecular tools to determine the origin of parasite infections in the animal trade (if infected before their export or import) is of great interest worldwide for both the animal trade industry and for animal welfare. Molecular tools have a wide range of applications, including forensic identification, wildlife preservation and conservation, veterinary public health protection, and food safety. Nonetheless, genetic markers were not reported to detect the source of infection in the animal trade. In this study we tested the applicability of molecular tools to detect the origin of Sarcoptes mite infection of wildebeest imported by the United Arab Emirate (UAE) from Tanzania. Using one multiplex of seven microsatellite markers and control samples from UAE, Kenya and Italy, we demonstrated the usefulness of the multiplex STR-typing as a molecular tool of pivotal interest to help commercialist, authorities, and conservationists, to identify the geographical origin of parasitic infection

First report of Setaria tundra in roe deer (Capreolus capreolus) from the Iberian Peninsula inferred from molecular data: epidemiological implications

Background Filarioid nematode parasites are major health hazards with important medical, veterinary and economic implications. Recently, they have been considered as indicators of climate change. Findings In this paper, we report the first record of Setaria tundra in roe deer from the Iberian Peninsula. Adult S. tundra were collected from the peritoneal cavity during the post-mortem examination of a 2 year-old male roe deer, which belonged to a private fenced estate in La Alcarria (Guadalajara, Spain). Since 2012, the area has suffered a high roe deer decline rate (75 %), for unknown reasons. Aiming to support the morphological identification and to determine the phylogenetic position of S. tundra recovered from the roe deer, a fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene from the two morphologically identified parasites was amplified, sequenced and compared with corresponding sequences of other filarioid nematode species. Phylogenetic analyses revealed that the isolate of S. tundra recovered was basal to all other formely reported Setaria tundra sequences. The presence of all other haplotypes in Northern Europe may be indicative of a South to North outbreak in Europe. Conclusions This is the first report of S. tundra in roe deer from the Iberian Peninsula, with interesting phylogenetic results, which may have further implications in the epidemiological and genetic studies of these filarioid parasites. More studies are needed to explore the reasons and dynamics behind the rapid host/geographic expansion of the filarioid parasites in EuropeThis work was supported by the Programme for Consolidating and Structuring Competitive Research Groups (GRC2015/003, Xunta de Galicia). Molecular analyses were carried out in the LEM of EBD, CSIC and funded by RNM 118; Junta AndaluciaS