DNA barcoding and morphology revealed the existence of seven new species of squat lobsters in the family Munididae (Decapoda, Galatheoidea) in the southwestern Pacific.

Specimens of squat lobsters belonging to the family MunididaeAhyong et al., 2010, representing the genera Garymunida Macpherson & Baba, 2022, Trapezionida Macpherson & Baba, 2022 and Typhlonida Macpherson & Baba, 2022, were collected during several cruises around New Caledonia and Papua New Guinea, Southwest Pacific. The integrative study of these specimens revealed the presence of one new species in Garymunida, five in Trapezionida and one in Typhlonida. We describe and illustrate these new species, providing some new data on the taxonomy of several rare or scarcely studied species of Trapezionida. Molecular data from different markers (mitochondrial and nuclear) was also included, based on data availability, to support the taxonomic status of different species. Finally, a key to species for each genus is also provided.

On the conservation of white-clawed crayfish in the Iberian Peninsula: Unraveling its genetic diversity and structure, and origin.

European crayfish species are a clear example of the drastic decline that freshwater species are experiencing. In particular, the native species of the Iberian Peninsula, the white clawed-crayfish (WCC) Austropotamobius pallipes, is listed as "endangered" by the IUCN and included in Annex II of the EU Habitat Directive and requires especially attention. Currently, implemented conservation management strategies require a better understanding of the genetic diversity and phylogeographic patterns, as well as of its evolutionary history. For this purpose, we have generated the largest datasets of two informative ribosomal mitochondrial DNA regions, i.e., cytochrome oxidase subunit I and 16S, from selected populations of the WCC covering its geographical distribution. These datasets allowed us to analyze in detail the (i) genetic diversity and structure of WCC populations, and (ii) divergence times for Iberian populations by testing three evolutionary scenarios with different mtDNA substitution rates (low, intermediate, and high rates). The results indicate high levels of haplotype diversity and a complex geographical structure for WCC in the Iberian Peninsula. The diversity found includes new unique haplotypes from the Iberian Peninsula and reveals that most of the WCC genetic variability is concentrated in the northern and central-eastern regions. Despite the fact that molecular dating analyses provided divergence times that were not statistically supported, the proposed scenarios were congruent with previous studies, which related the origin of these populations with paleogeographic events during the Pleistocene, which suggests an Iberian origin for these WCC. All results generated in this study, indicate that the alternative hypothesis of an introduced origin of the Iberian WCC is highly improbable. The result of this study, therefore, has allowed us to better understand of the genetic diversity, structure patterns, and evolutionary history of the WCC in the Iberian Peninsula, which is crucial for the management and conservation needs of this endangered species.

A horizon scan exercise for aquatic invasive alien species in Iberian inland waters.

As the number of introduced species keeps increasing unabatedly, identifying and prioritising current and potential Invasive Alien Species (IAS) has become essential to manage them. Horizon Scanning (HS), defined as an exploration of potential threats, is considered a fundamental component of IAS management. By combining scientific knowledge on taxa with expert opinion, we identified the most relevant aquatic IAS in the Iberian Peninsula, i.e., those with the greatest geographic extent (or probability of introduction), severe ecological, economic and human health impacts, greatest difficulty and acceptability of management. We highlighted the 126 most relevant IAS already present in Iberian inland waters (i.e., Concern list) and 89 with a high probability of being introduced in the near future (i.e., Alert list), of which 24 and 10 IAS, respectively, were considered as a management priority after receiving the highest scores in the expert assessment (i.e., top-ranked IAS). In both lists, aquatic IAS belonging to the four thematic groups (plants, freshwater invertebrates, estuarine invertebrates, and vertebrates) were identified as having been introduced through various pathways from different regions of the world and classified according to their main functional feeding groups. Also, the latest update of the list of IAS of Union concern pursuant to Regulation (EU) No 1143/2014 includes only 12 top-ranked IAS identified for the Iberian Peninsula, while the national lists incorporate the vast majority of them. This fact underlines the great importance of taxa prioritisation exercises at biogeographical scales as a step prior to risk analyses and their inclusion in national lists. This HS provides a robust assessment and a cost-effective strategy for decision-makers and stakeholders to prioritise the use of limited resources for IAS prevention and management. Although applied at a transnational level in a European biodiversity hotspot, this approach is designed for potential application at any geographical or administrative scale, including the continental one.

House sparrows do not exhibit a preference for the scent of potential partners with different MHC-I diversity and genetic distances.

MHC genes play a fundamental role in immune recognition of pathogens and parasites. Therefore, females may increase offspring heterozygosity and genetic diversity by selecting males with genetically compatible or heterozygous MHC. In birds, several studies suggest that MHC genes play a role in mate choice, and recent evidence suggests that olfaction may play a role in the MHC-II discrimination. However, whether olfaction is involved in MHC-I discrimination in birds remains unknown. Previous studies indicate that house sparrow females with low allelic diversity prefer males with higher diversity in MHC-I alleles. Here, we directly explored whether female and male house sparrows (Passer domesticus) could estimate by scent MHC-I diversity and/or dissimilarity of potential partners. Our results show that neither females nor males exhibit a preference related to MHC-I diversity or dissimilarity of potential partners, suggesting that MHC-I is not detected through olfaction. Further studies are needed to understand the mechanisms responsible for mate discrimination based on MHC-I in birds.

The shrunk genetic diversity of coral populations in North-Central Patagonia calls for management and conservation plans for marine resources.

The Chilean Patagonia is a complex puzzle of numerous fjords, channels, bays, estuaries, and islands. The largest part of it is very remote, hampering the generation of scientific knowledge and effective management planning that could balance conservation of the marine resources with the increasing development of aquaculture activities. The present study focuses on the deep-water emergent cold-water coral Desmophyllum dianthus, dwelling in Chilean Patagonia, with the aim to illustrate its population genetic structure, demography and adaptation of the species along this coast. Microsatellite loci analysis included D. dianthus individuals from twelve sampling localities along bathymetric and oceanographic gradients from the latitude 40°S to 48°S. The results showed a lack of genetic structure with an asymmetric dispersion of individuals, and relevant heterozygosity deficiency in some populations. This study also analyses the natural and human impacts affecting the region (e.g., climate change, increasing salmon farming activities), and stresses the importance of including genetic information in the process of management and conservation of marine resources. In particular, the relevance of using interdisciplinary approaches to fill the gaps in scientific knowledge especially in remote and pristine areas of western Patagonia. Therefore, information on genetic spatial distribution of marine fauna could become pivotal to develop a holistic ecosystem-based approach for marine spatial planning.

Genetic Structure of the Endangered Coral Cladocora caespitosa Matches the Main Bioregions of the Mediterranean Sea.

Population connectivity studies are a useful tool for species management and conservation planning, particular of highly threatened or endangered species. Here, we evaluated the genetic structure and connectivity pattern of the endangered coral Cladocora caespitosa across its entire distribution range in the Mediterranean Sea. Additionally, we examined the relative importance of sexual and asexual reproduction in the studied populations and their genetic diversity. A total of 541 individuals from 20 localities were sampled and analysed with 19 polymorphic microsatellite markers. Of the genotyped individuals, 482 (89%) had unique multilocus genotypes. Clonality percentages of the populations varied from 0% (in eight populations) to nearly 69% (in one population from Crete). A heterozygosity deficit and a high degree of inbreeding was the general trend in our data set. Population differentiation in C. caespitosa was characterised by significant pairwise F ST values with lower ones observed at an intraregional scale and higher ones, between populations from different biogeographic regions. Genetic structure analyses showed that the populations are divided according to the three main sub-basins of the Mediterranean Sea: the Western (Balearic, Ligurian and Tyrrhenian seas), the Central (Adriatic and Ionian seas) and the Eastern (Levantine and Aegean seas), coinciding with previously described gene flow barriers. However, the three easternmost populations were also clearly separated from one another, and a substructure was observed for the other studied areas. An isolation-by-distance pattern was found among, but not within, the three main population groups. This substructure is mediated mainly by dispersal along the coastline and some resistance to larval movement through the open sea. Despite the low dispersal ability and high self-recruitment rate of C. caespitosa, casual dispersive events between regions seem to be enough to maintain the species' considerable genetic diversity. Understanding the population connectivity and structure of this endangered scleractinian coral allows for more informed conservation decision making.

Climate warming triggers the emergence of native viruses in Iberian amphibians.

The number of epizootics in amphibian populations caused by viruses of the genus Ranavirus is increasing worldwide. Yet, causes for pathogen emergence are poorly understood. Here, we confirmed that the Common midwife toad virus (CMTV) and Frog virus 3 (FV3) are responsible for mass mortalities in Iberia since the late 1980s. Our results illustrate the Iberian Peninsula as a diversity hotspot for the highly virulent CMTV. Although this pattern of diversity in Europe is consistent with spread by natural dispersal, the exact origin of the emergence of CMTV remains uncertain. Nevertheless, our data allow hypothesizing that the Iberian Peninsula might harbor the ancestral population of CMTVs that could have spread into the rest of Europe. In addition, we found that climate warming could be triggering the CMTV outbreaks, supporting its endemic status in the Iberian Peninsula.

Revision of the squat lobsters of the genus Phylladiorhynchus Baba, 1969 (Crustacea, Decapoda, Galatheidae) with the description of 41 new species.

The genus Phylladiorhynchus Baba, 1969 currently contains 11 species, all occurring in the shallow waters and on the continental shelf of the Indian and Pacific oceans. Recent expeditions in these oceans have resulted in the collection of numerous new specimens in need of analysis. We have studied this material using an integrative approach analysing both morphological and molecular (COI and 16S) characters. We describe 41 new species and resurrect three old names: P. integrus (Benedict, 1902) and P. lenzi (Rathbun, 1907), previously synonymized with P. pusillus (Henderson, 1885), and P. serrirostris (Melin, 1939), previously synonymized with P. integrirostris (Dana, 1852). Most species of the genus are described and illustrated. Some species are barely discernible on the basis of morphological characters but are highly divergent genetically. Species of Phylladiorhynchus are mainly distinguishable by the number of epigastric spines and lateral spines of the carapace, the shape and the armature of the rostrum, the number and pattern of the ridges on the carapace and pleon, the shape of thoracic sternite 3 and the armature of the P24 dactyli. A dichotomous identification key to all species is provided.

Genetic connectivity of two marine gastropods in the Mediterranean Sea: seascape genetics reveals species-specific oceanographic drivers of gene flow.

Oceanographic features such as currents, waves, temperature and salinity, together with life history traits, control patterns and rates of gene flow and contribute to shaping the population genetic structure of marine organisms. Seascape genetics is an emerging discipline that adopts a spatially explicit approach to examine biotic and abiotic factors that drive gene flow in marine environments. In this study, we examined factors that contribute to genetic differentiation in two coastal Mediterranean gastropods whose geographical ranges overlap but which inhabit different environments. The two species differ in several life history traits and in their dispersal capabilities. Genetic differentiation was relatively low for the trochid species Gibbula divaricata (FST  =0.059), and high for the vermetid species Dendropoma lebeche (FST  =0.410). Salinity emerged as the most important variable explaining the genetic structure of both species; sea surface temperature was also important for G. divaricata. For the more sessile D. lebeche, the coastline was predicted to provide important pathways for stepping-stone connectivity and gene flow. Our results provide a greater understanding of the factors influencing marine population connectivity, which may be useful to guide marine conservation and management in the Mediterranean.

Asexual reproduction in bad times? The case of Cladocora caespitosa in the eastern Mediterranean Sea.

We analysed the patterns of genetic variability of eastern Mediterranean populations of the scleractinian coral Cladocora caespitosa, from the Aegean and Levantine seas, using 19 polymorphic microsatellite loci, 11 of which were newly characterized. The observed genetic pattern reflects a scenario of isolation by environment: F ST comparisons showed a higher degree of genetic differentiation between the two Cypriot populations that are separated by only 11 km than between these two Levantine populations and the Aegean population in Greece, which are separated by 1300 km. We hypothesize that local-scale oceanographic factors influenced the dispersal of planulae between the geographically close populations, playing a crucial role in the genetic structure of this coastal coral. Yet, despite being characterized as a species with limited dispersal and high self-recruitment, large-scale migration does eventually occur as first-generation migrants were identified between the most distant populations. In line with previous findings of reproductive plasticity in C. caespitosa, we also found localized differences in reproduction mode (sexual vs. asexual) within a geographically limited context. Several individuals were identified as clones, indicating the predominance of asexual reproduction in one of the Cypriot populations. We interpret this predominance either as a direct response to or as an indirect consequence of perturbations suffered by this C. caespitosa population. These perturbations are caused by unfavourable environmental conditions that threatened local survival, in particular water temperature changes and windstorm swells. Asexual reproduction may be a mechanism used by C. caespitosa to counteract mortality events and recolonize devastated areas, and likely accounts for the occasional high levels of clonality and low levels of genetic diversity. Local adaptations such as these should therefore be considered in conservation and management strategies to maintain and preserve the gene pool of this endangered species. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s00338-020-02040-3).

New occurrences of squat lobsters of the genus Eumunida Smith, 1883 (Decapoda, Eumunididae) in New Caledonia, the Solomon Islands and Papua-New Guinea, with the description of a new species.

Examination of numerous specimens of squat lobsters of the genus Eumunida Smith, 1883 collected by French cruises along the coasts of New Caledonia, the Solomon Islands and Papua-New Guinea revealed the presence of six species, including a new species. The collection data of all of these species are recorded. The new species, E. turbulenta n. sp., is described and illustrated from New Caledonia and Chesterfield Islands.

Aproximación morfológica y molecular al conocimiento de las ofiuras (Echinodermata: Ophiuroidea) en el Parque Nacional Marino Las Baulas (Pacífico Norte, Costa Rica)

Introducción: El conocimiento de la Clase Ophiuroidea es muy escaso y deficiente, a pesar de ser la que presenta el mayor número de especies del Filo Echinodermata con hasta 2 100. Además, apenas existen publicaciones dedicadas específicamente al grupo de las ofiuras en la vertiente pacífica de Costa Rica. Objetivo: La caracterización morfológica y genética de las ofiuras de la franja intermareal y sublitoral somero del Parque Nacional Marino Las Baulas (Pacífico de Costa Rica). Métodos: El periodo de recolecta se extendió entre enero y abril de 2018 y tuvo lugar a lo largo de siete localidades: Punta del Morro, Playa Carbón, Playa Ventanas, Playa Grande, Playa Tamarindo Ciudad, Playa Tamarindo y Playa Langosta. Se ha realizado la caracterización de 214 ofiuras mediante claves taxonómicas, descripciones originales y el marcador mitocondrial citocromo c oxidasa subunidad I (COI). Resultados: En total, combinando los caracteres morfológicos y genéticos, se han identificado 11 especies, cuatro de las cuales podrían ser nuevas, nueve constituyen nuevos registros en el parque y una de las especies se ha encontrado por primera vez en Costa Rica (Ophiophragmus aff. stellatus). Además, se han identificado individuos juveniles y se han encontrado dos especies crípticas. Conclusión: Del estudio molecular se concluye que el marcador COI es útil para delimitar molecularmente las especies y establecer relaciones filogenéticas para los niveles taxonómicos de género y especie, pero no para los superiores. Se aportan observaciones morfológicas e imágenes como contribución a la taxonomía del grupo.

Introduction: Information on the class Ophiuroidea is very scarce, despite it consisting of the highest number of species within the phylum Echinodermata, with up to 2 100 species. In particular, there are hardly any study focused specifically to ophiuroids found along the Pacific coast of Costa Rica. Objective: Morphological and genetic characterization of ophiuroids of the shallow intertidal and sublittoral areas of Las Baulas Marine National Park (Pacific coast of Costa Rica). Methods: Sampling was conducted between January and April of 2018 at seven locations: Punta del Morro, Playa Carbón, Playa Ventanas, Playa Grande, Playa Tamarindo Ciudad, Playa Tamarindo and Playa Langosta. A total of 214 ophiuroids were characterized using taxonomic keys and original descriptions and a molecular analysis of the mitochondrial Cytochrome oxidase I (COI) gene. Results: An analysis of morphological and genetic characters indicates that 11 species are present in the study site. Four may represent new species, nine constitute new records for the area and one (Ophiophragmus aff. stellatus) is recorded in Costa Rica for the first time. Additionally, juvenile individuals of different species were identified, and two cryptic species were detected. Conclusions: According to the molecular analysis, COI is useful to delimit ophiuroid species and to establish phylogenetic relationships at lower (genera and species) but not higher taxonomic levels. This study also provides images and morphological remarks for future comparative studies.

A new species of squat lobster of the genus Hendersonida (Crustacea, Decapoda, Munididae) from Papua New Guinea.

Hendersonida parvirostris sp. nov. is described from Papua New Guinea. The new species can be distinguished from the only other species of the genus, H. granulata (Henderson, 1885), by the fewer spines on the dorsal carapace surface, the shape of the rostrum and supraocular spines, the antennal peduncles, and the length of the walking legs. Pairwise genetic distances estimated using the 16S rRNA and COI DNA gene fragments indicated high levels of sequence divergence between the new species and H. granulata. Phylogenetic analyses, however, recovered both species as sister species, supporting monophyly of the genus.

Revision of the squat lobsters of the genus Leiogalathea Baba, 1969 (Crustacea, Decapoda, Munidopsidae) with the description of 15 new species.

The genus Leiogalathea Baba, 1969 currently contains only two benthic species both occurring on the continental shelves and slope: L. laevirostris (Balss, 1913), widely reported in the Indo-Pacific region, and L. agassizii (A. Milne Edwards, 1880), from both sides of the Central Atlantic. A certain degree of morphological variability linked to their geographic distributions was previously noticed, mostly in L. laevirostris. In the present study, we revise numerous specimens collected from the Atlantic, Indian and Pacific Oceans, analysing morphological and molecular characters (COI and 16S rRNA). We found 15 new species; all of them are distinguished from L. laevirostris and L. agassizii by subtle but constant morphological differences and show clear genetic separation. Furthermore, L. imperialis (Miyake Baba, 1967), previously synonymized with L. laevirostris, was found to be a valid species. All species are described and illustrated. Species of the genus Leiogalathea are morphologically distinguishable on the basis of the spinulation of the carapace, the shape and the armature of the rostrum, the shape of the propodi of the walking legs, and the pattern of the setae covering on rostrum, carapace and chelae. Some species are barely discernible on the basis of these characters but are highly divergent genetically.

Connectivity Among Populations of the Top Shell Gibbula divaricata in the Adriatic Sea.

Genetic connectivity studies are essential to understand species diversity and genetic structure and to assess the role of potential factors affecting connectivity, thus enabling sound management and conservation strategies. Here, we analyzed the patterns of genetic variability in the marine snail Gibbula divaricata from five coastal locations in the central-south Adriatic Sea (central Mediterranean) and one in the adjacent northern Ionian Sea, using 21 described polymorphic microsatellite loci. Observed and expected heterozygosity varied from 0.582 to 0.635 and 0.684 to 0.780, respectively. AMOVA analyses showed that 97% of genetic variation was observed within populations. Nevertheless, significant, although small, genetic differentiation was found among nearly all of the pairwise F ST comparisons. Over a general pattern of panmixia, three groups of populations were identified: eastern Adriatic populations, western Adriatic populations, and a third group represented by the single northern Ionian Sea population. Nonetheless, migration and gene flow were significant between these groups. Gibbula divaricata is thought to have a limited dispersal capacity related to its lecithotrophic trochophore larval stage. Our results indicated high levels of self-recruitment and gene flow that is mainly driven through coastline dispersion, with populations separated by the lack of suitable habitats or deep waters. This stepping-stone mode of dispersion together with the high levels of self-recruitment could lead to higher levels of population structuring and differentiation along the Adriatic Sea. Large effective population sizes and episodic events of long-distance dispersal might be responsible for the weak differentiation observed in the analyzed populations. In summary, the circulation system operating in this region creates natural barriers for dispersion that, together with life-history traits and habitat requirements, certainly affect connectivity in G. divaricata. However, this scenario of potential differentiation seems to be overridden by sporadic events of long-distance dispersal across barriers and large effective population sizes.

Three new species of squat lobsters of the genus Munidopsis Whiteaves, 1874, from Guadeloupe Island, Caribbean Sea (Crustacea, Decapoda, Munidopsidae).

The genus Munidopsis is one of the most diverse genera within squat lobsters. Here, three new species of Munidopsis, M. cornuata n. sp., M. senticosa n. sp., and M. turgida n. sp., from <500 m off Guadeloupe Island (Caribbean Sea), are fully described and illustrated. Among the Atlantic species of the genus, M. cornuata n. sp. belongs to the group of species having the dorsal surface of the carapace with spines and is most similar to M. robusta (A. Milne-Edwards, 1880), from the Gulf of Mexico and Caribbean Sea. Munidopsis senticosa n. sp. resembles M. barbarae (Boone, 1927) from the Bahamas and the Gulf of Mexico and M. penescabra Pequegnat Williams 1995, from off Georgia and Gulf of Mexico; the three species belong to the group having the carapace covered with sharp spines. Finally, M. turgida n. sp. is characterized by having the dorsal surface of the carapace, abdomen and pereiopods covered by granules; and resembles M. granulens Mayo, 1972, from NW Caribbean Sea. Apart from the morphological evidence, the analysis of mitochondrial genes (COI and 16S) supports establishing these new species, showing very high genetic divergences compared to their congeners (from 14.5 to 17% for COI, and 7.7 to 12.8% for 16S data).

Isolation of microsatellite loci for the endangered vermetid gastropod Dendropoma lebeche using Illumina MiSeq next generation sequencing technology.

Dendropoma petraeum, considered the primary vermetid reef-building species in the Mediterranean, has recently been shown to be a species complex of at least four cryptic species. These species have highly restricted, non-overlapping distributions, causing concern for their conservation status. To better study the genetic diversity of these populations, we selected one of these species, Dendropoma lebeche (Templado et al. in Mediterr Mar Sci 17(1):13-31, 2016), which is restricted to the western Mediterranean, for microsatellite marker development using Illumina MiSeq. We provide an initial survey of 29 polymorphic microsatellite loci for D. lebeche. Genetic analyses identified 2-11 alleles per locus across the 30 samples examined. Observed and expected heterozygosities ranged from 0.067 to 0.800 and 0.064 to 0.770, respectively. None of the loci deviated from Hardy-Weinberg equilibrium or showed signs of being under selective pressure. Significant linkage disequilibrium was found between two loci. We also show the cross-species amplification of these microsatellite markers in the other three species of the complex, the Tyrrhenian-Sicilian lineage, D. cristatum (Biondi, 1859), the Levantine lineage, D. anguliferum (Monterosato, 1878) and Dendropoma sp. found along the Ionian-Aegean coasts, suggesting their potential utility for future phylogenetic and evolutionary studies.

Predatory flying squids are detritivores during their early planktonic life.

Cephalopods are primarily active predators throughout life. Flying squids (family Ommastrephidae) represents the most widely distributed and ecologically important family of cephalopods. While the diets of adult flying squids have been extensively studied, the first feeding diet of early paralarvae remains a mystery. The morphology of this ontogenetic stage notably differs from other cephalopod paralarvae, suggesting a different feeding strategy. Here, a combination of Laser Capture Microdissection (LCM) and DNA metabarcoding of wild-collected paralarvae gut contents for eukaryotic 18S v9 and prokaryotic 16S rRNA was applied, covering almost every life domain. The gut contents were mainly composed by fungus, plants, algae and animals of marine and terrestrial origin, as well as eukaryotic and prokaryotic microorganisms commonly found in fecal pellets and particulate organic matter. This assemblage of gut contents is consistent with a diet based on detritus. The ontogenetic shift of diet from detritivore suspension feeding to active predation represents a unique life strategy among cephalopods and allows ommastrephid squids to take advantage of an almost ubiquitous and accessible food resource during their early stages. LCM was successfully applied for the first time to tiny, wild-collected marine organisms, proving its utility in combination with DNA metabarcoding for dietary studies.