Genomic Evidence for Cryptic Speciation in Tree Frogs From the Apennine Peninsula, With Description of Hyla perrini sp. nov

Despite increasing appreciation of the speciation continuum, delimiting and describing new species is a major yet necessary challenge of modern phylogeography to help optimize conservation efforts. In amphibians, the lack of phenotypic differences between closely-related taxa, their complex, sometimes unresolved phylogenetic relationships, and their potential to hybridize all act to blur taxonomic boundaries. Here we implement a multi-disciplinary approach to evaluate the nature of two deeply-diverged mitochondrial lineages previously documented in Italian tree frogs (Hyla intermedia s. l.), distributed north and south of the Northern Apennine Mountains. Based on evidence from mitochondrial phylogenetics, nuclear phylogenomics, hybrid zone population genomics, niche modeling analyses, and biometric assessments, we propose that these lineages be considered distinct, cryptic species. Both mitochondrial and nuclear data affirm that they belong to two monophyletic clades of Pliocene divergence (~3.5 My), only admixing over a relatively narrow contact zone restricted to the southeast of the Po Plain (50–100 km). These characteristics are comparable to similarly-studied parapatric amphibians bearing a specific status. Inferred from their current geographic distribution, the two Italian tree frogs feature distinct ecological niches (<15% of niche overlap), raising questions regarding potential adaptive components contributing to their incipient speciation. However, we found no diagnostic morphological and bioacoustic differences between them. This system illustrates the speciation continuum of Western-Palearctic tree frogs and identifies additional cryptic lineages of similar divergence to be treated as separate species (H. cf. meridionalis). We recommend combined approaches using genomic data as applied here for the future taxonomic assessment of cryptic diversity in alloparapatric radiations of terrestrial vertebrates, especially in controversial taxa. Finally, we formally described the northern Italian tree frogs as a new species, Hyla perrini sp. nov

Bi-allelic Loss-of-Function CACNA1B Mutations in Progressive Epilepsy-Dyskinesia.

The occurrence of non-epileptic hyperkinetic movements in the context of developmental epileptic encephalopathies is an increasingly recognized phenomenon. Identification of causative mutations provides an important insight into common pathogenic mechanisms that cause both seizures and abnormal motor control. We report bi-allelic loss-of-function CACNA1B variants in six children from three unrelated families whose affected members present with a complex and progressive neurological syndrome. All affected individuals presented with epileptic encephalopathy, severe neurodevelopmental delay (often with regression), and a hyperkinetic movement disorder. Additional neurological features included postnatal microcephaly and hypotonia. Five children died in childhood or adolescence (mean age of death: 9 years), mainly as a result of secondary respiratory complications. CACNA1B encodes the pore-forming subunit of the pre-synaptic neuronal voltage-gated calcium channel Cav2.2/N-type, crucial for SNARE-mediated neurotransmission, particularly in the early postnatal period. Bi-allelic loss-of-function variants in CACNA1B are predicted to cause disruption of Ca2+ influx, leading to impaired synaptic neurotransmission. The resultant effect on neuronal function is likely to be important in the development of involuntary movements and epilepsy. Overall, our findings provide further evidence for the key role of Cav2.2 in normal human neurodevelopment.MAK is funded by an NIHR Research Professorship and receives funding from the Wellcome Trust, Great Ormond Street Children's Hospital Charity, and Rosetrees Trust. E.M. received funding from the Rosetrees Trust (CD-A53) and Great Ormond Street Hospital Children's Charity. K.G. received funding from Temple Street Foundation. A.M. is funded by Great Ormond Street Hospital, the National Institute for Health Research (NIHR), and Biomedical Research Centre. F.L.R. and D.G. are funded by Cambridge Biomedical Research Centre. K.C. and A.S.J. are funded by NIHR Bioresource for Rare Diseases. The DDD Study presents independent research commissioned by the Health Innovation Challenge Fund (grant number HICF-1009-003), a parallel funding partnership between the Wellcome Trust and the Department of Health, and the Wellcome Trust Sanger Institute (grant number WT098051). We acknowledge support from the UK Department of Health via the NIHR comprehensive Biomedical Research Centre award to Guy's and St. Thomas' National Health Service (NHS) Foundation Trust in partnership with King's College London. This research was also supported by the NIHR Great Ormond Street Hospital Biomedical Research Centre. J.H.C. is in receipt of an NIHR Senior Investigator Award. The research team acknowledges the support of the NIHR through the Comprehensive Clinical Research Network. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, Department of Health, or Wellcome Trust. E.R.M. acknowledges support from NIHR Cambridge Biomedical Research Centre, an NIHR Senior Investigator Award, and the University of Cambridge has received salary support in respect of E.R.M. from the NHS in the East of England through the Clinical Academic Reserve. I.E.S. is supported by the National Health and Medical Research Council of Australia (Program Grant and Practitioner Fellowship)

Evolution of asexuality and maintenance of reproductive polymorphism in thrips (Thysanoptera)

Why do the majority of eukaryotes favor the complex and resource-intensive route of sexual reproduction, despite the availability of simpler and cheaper alternatives like asexuality? Reproduction is one of the most fundamental aspects of life; and this question continues to be a central and largely unresolved query in the field of evolutionary biology as the mechanisms through which sexual reproduction is maintained in nature remain unknown. The present work aims to resolve this evolutionary mystery, employing grass thrips belonging to the genus Aptinothrips as a study system. This group of herbivorous insects, native to Europe, provides an interesting case for exploring and contrasting sexual and asexual reproduction. Three independently derived asexual lineages have been identified within this group, each originating from a sexual lineage, with one asexual species being particularly diverse and competitive, Aptinothrips rufus. Females of this species are endosymbiont-induced parthenogens infected by Wolbachia, which might play an important role in generating diversity in this species, a key component of its apparent success compared to its relatives. In natural populations, sexual individuals and their parthenogenetic counterparts are frequently coexisting, yet there is currently not a single system in which the persistence of reproductive polymorphism can be explained. The coexistence of sexual and asexual grass thrips in the same meadows offers an ideal opportunity to investigate the factors contributing to the maintenance of both reproductive modes when they are in promiscuity. In this work, I firstly assessed the role played by Wolbachia in the generation of clonal lineages within the genus Aptinothrips with a focus on the very diverse asexual Aptinothrips rufus. This study then explores whether the clonal diversity observed in asexuals is functionally relevant, specifically whether it results in niche differentiation among genetically distinct asexual lineages. Subsequently, both experimental setups and natural settings are employed to measure the ecological niches of sexual and asexual populations, aiming to assess whether variations in niche size and types contribute to the coexistence of both reproductive strategies in wild populations. Simultaneously, ecological correlates with sex are examined, particularly in relation to enhanced habitat heterogeneity, as theoretical predictions suggest an advantage of sexual reproduction in such circumstances. Ultimately, we perform an additional evaluation of the reproductive mode in the sexual species A. elegans, noting a limited production of males in the Swiss Prealps, and discuss potential implications for future studies. -- Pourquoi la majorité des eucaryotes privilégient-ils la voie complexe et coûteuse de la reproduction sexuée, malgré l'existence d'alternatives plus simples et moins coûteuses comme l'asexualité ? La reproduction est l'un des aspects les plus fondamentaux de la vie ; et cette question demeure une interrogation centrale et largement non résolue dans le domaine de la biologie évolutive, car les mécanismes permettant le maintien de la reproduction sexuée dans la nature restent inconnus. Le présent travail vise à résoudre ce mystère évolutif, en utilisant les thrips du genre Aptinothrips comme système d'étude. Ce groupe d'insectes herbivores, originaire d'Europe, offre un cas intéressant pour explorer et contraster la reproduction sexuée et asexuée. Trois lignées asexuées indépendamment dérivées ont été identifiées au sein de ce groupe, chacune provenant d'un ancêtre sexué, avec une espèce asexuée en particulier, Aptinothrips rufus, qui se distingue par sa diversité et sa compétitivité. Les femelles de cette espèce sont infectées par par Wolbachia, un endosymbiote induisant la parthénogénèse qui pourrait jouer un rôle important dans la génération de la diversité chez cette espèce, un élément clé de son succès apparent par rapport à ses congénères. Dans les populations naturelles, les individus sexués et leurs homologues parthénogénétiques coexistent fréquemment, mais il n'existe actuellement aucun système dans lequel la persistance du polymorphisme reproducteur puisse être expliquée. La coexistence de thrips sexués et asexués dans les mêmes prairies offre une occasion idéale d'investiguer les facteurs contribuant au maintien des deux modes de reproduction lorsqu'ils sont en promiscuité. Dans ce travail, j'ai d'abord évalué le rôle joué par Wolbachia dans la génération de lignées clonales au sein du genre Aptinothrips, en mettant l'accent sur l'Aptinothrips rufus. Elle explore ensuite, à l’aide de dispositifs expérimentaux, puis dans l’environnement naturel, si des différences de niche écologique existent entre différents clones et entre différentes espèces sexuées ou asexuées. Les impacts de ces différences sur la coexistence des deux stratégies de reproduction sont ensuite discutés Parallèlement les conditions théoriques favorisant habituellement les espèces sexuées, comme une hétérogénéité accrue des habitats, ont été testées. Enfin, nous réalisons une évaluation supplémentaire du mode de reproduction dans l'espèce sexuée A. elegans, notant une production limitée de mâles dans les Préalpes suisses, et discutons des implications potentielles pour les études futures

Drivers of Patients’ Behavioral Intention toward Public and Private Clinics’ Services

In an era of growing competition in the healthcare market, adopting a patient-centered approach is mandatory for the survival and growth of any public or private hospital. This requires a better understanding of patients’ behavior and an increased focus on satisfying their needs and expectations. This paper was developed in this context and aims to study the main drivers of patients’ behavioral intentions. A conceptual model was proposed, highlighting the linkages between service quality, doctors’ reputation, patients’ trust, service value, and patients’ behavioral intentions. To examine the different research hypotheses, a quantitative study including 242 patients was conducted in Saudi Arabia using the convenience sampling method. The smart PLS approach was used to test the measurement and structural models. The findings indicated that trust and service value positively affected patients’ behavioral intentions. Trust in the healthcare provider was positively affected by two dimensions of service quality: healthcare provider concern and physician concern. Trust in doctors was found to be positively related to the reputation of the doctor. Service value was positively influenced by the convenience of the healthcare process, healthcare provider concerns, and doctors’ reputations. This study is original because it is among the few studies that investigate patients’ behavioral intentions toward healthcare services in a developing country (Saudi Arabia). Furthermore, it is among the rare studies to examine the role of doctors’ reputations in service values. The findings would offer meaningful implications for practitioners in the healthcare market for maintaining relationships with their patients

Introgressive hybridization of threatened European tree frogs ( Hyla arborea ) by introduced H. intermedia in Western Switzerland

Hybridization by introduced taxa is a major threat to native species. Characterizing human introductions is thus one of the missions of conservation geneticists. Here we survey a declining population of the regionally endangered European tree frog (Hyla arborea) in the Grangettes natural reserve (Rhone valley, Western Switzerland), where previous evidence indicated human introduction of the Italian taxon Hyla intermedia. We combined fast-evolving mitochondrial and nuclear markers and an extended sampling to conduct population genetic analyses of the Grangettes and putative source areas. We show that the Grangettes population is a hybrid swarm, with all individuals featuring recent nuclear admixture and mitochondrial DNA of introduced H. intermedia, most likely of proximate south Alpine origin. In contrast, H. arborea and H. intermedia hardly introgress in their natural parapatric ranges, consistent with an advanced reproductive isolation. From our results, even distantly-related species can endanger the genetic integrity of local taxa in the context of introductions. Potential hybrid incompatibilities may thus account for the strong decline of this population, despite important conservation efforts. Although their hybrid nature makes them a priori unworthy of any protection, we propose specific measures to recover local H. arborea gene pool and preserve tree frogs in the Grangettes, the last population remaining from this heavily impacted part of the Alps

Data from: Evolutionary melting pots: a biodiversity hotspot shaped by ring diversifications around the Black Sea in the Eastern tree frog (Hyla orientalis)

Hotspots of intraspecific genetic diversity, which are of primary importance for the conservation of species, have been associated with glacial refugia, that is areas where species survived the Quaternary climatic oscillations. However, the proximate mechanisms generating these hotspots remain an open issue. Hotspots may reflect the long-term persistence of large refugial populations; alternatively, they may result from allopatric differentiation between small and isolated populations, that later admixed. Here, we test these two scenarios in a widely distributed species of tree frog, Hyla orientalis, which inhabits Asia Minor and southeastern Europe. We apply a fine-scale phylogeographic survey, combining fast-evolving mitochondrial and nuclear markers, with a dense sampling throughout the range, as well as ecological niche modelling, to understand what shaped the genetic variation of this species. We documented an important diversity centre around the Black Sea, composed of multiple allopatric and/or parapatric diversifications, likely driven by a combination of Pleistocene climatic fluctuations and complex regional topography. Remarkably, this diversification forms a ring around the Black Sea, from the Caucasus through Anatolia and eastern Europe, with terminal forms coming into contact and partially admixing in Crimea. Our results support the view that glacial refugia generate rather than host genetic diversity and can also function as evolutionary melting pots of biodiversity. Moreover, we report a new case of ring diversification, triggered by a large, yet cohesive dispersal barrier, a very rare situation in nature. Finally, we emphasize the Black Sea region as an important centre of intraspecific diversity in the Palearctic with implications for conservation

Fungal infection in free-ranging snakes caused by opportunistic species

Emerging infectious fungal diseases such as chytridiomycosis, caused by Batrachochytrium dendrobatidis and B. salamandrivorans in amphibians, or ophidiomycosis in reptiles (Ophidiomyces ophiodiicola), are major threats that can drive species or entire populations to extinction over a short period of time. Besides these well-documented pathogens, such diseases can be caused by numerous opportunistic fungal species that can target e.g. immunocompromised individuals from various species of vertebrates. In reptiles, opportunistic species are mainly documented in captive animals kept in inadequate conditions, but data remains scarce for wild individuals. In the present study, we isolated and genetically identified fungal species responsible of skin lesions in wild-caught smooth snakes (Coronella austriaca) during a field survey targeting endangered reptile species in Switzerland. A total of 18 fungal species were isolated and genetically identified from the lesions of the two wild-caught snakes and included several species known for being opportunistic pathogens in vertebrates and infecting mainly immunocompromised individuals, such as Alternaria infectoria and Rhodotorula spp. It is not possible to establish whether the snakes had such an issue. However, the exceptional wet and cold conditions experienced in spring 2021 might have trigger the infections. Indeed, high humidity has been recorded as a predisposing factor for mycoses in captive reptiles

Sequence alignment of cytochrome-b

This includes 238 samples of H. arborea and 2 samples of H. orientali