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.

An evaluation of errors in the mitochondrial COI sequences of Hydrachnidia (Acari, Parasitengona) in public databases.

Public molecular databases are fundamental tools for modern taxonomic studies whose usefulness rely on the soundness of the data within them. Here, we study potential errors that can arise along the data pipeline from sampling, specimen identification and molecular processing (digestion, amplification and sequencing) to the submission of sequences to these databases by using the DNA sequences of Hydrachnidia (Acari, Parasitengona) as a case study. Our results indicate that molecular information is available for only about 3% of the Hydrachnidia species known to date; yet, within this small percentage, errors are present in almost 5% of the species analyzed (0.5% of the sequences and almost 11% of the genera). This study underscores the scarcity of genetic data available for Hydrachnidia, but also that the proportion of errors in DNA sequences is relatively small. Even so, it highlights the danger associated with using DNA sequences from public databases, particularly for species identification, and reinforces the need for greater quality control measures and/or protocols to avoid an intensification of errors in the (post) genomics era. Finally, our study emphasizes that potential errors may also reveal cryptic diversity within a species.

Molecular Correlation between Larval, Deutonymph and Adult Stages of the Water Mite Arrenurus (Micruracarus) Novus.

The systematics of many groups of organisms has been based on the adult stage. Morphological transformations that occur during development from the embryonic to the adult stage make it difficult (or impossible) to identify a juvenile (larval) stage in some species. Hydrachnidia (Acari, Actinotrichida, which inhabit mainly continental waters) are characterized by three main active stages-larval, deutonymph and adult-with intermediate dormant stages. Deutonymphs and adults may be identified through diagnostic morphological characters. Larvae that have not been tracked directly from a gravid female are difficult to identify to the species level. In this work, we compared the morphology of five water mite larvae and obtained the molecular sequences of that found on a pupa of the common mosquito Culex (Culex) pipiens with the sequences of 51 adults diagnosed as Arrenurus species and identified the undescribed larvae as Arrenurus (Micruracarus) novus. Further corroborating this finding, adult A. novus was found thriving in the same mosquito habitat. We established the identity of adult and deutonymph A. novus by morphology and by correlating COI and cytB sequences of the water mites at the larval, deutonymph and adult (both male and female) life stages in a particular case of 'reverse taxonomy'. In addition, we constructed the Arrenuridae phylogeny based on mitochondrial DNA, which supports the idea that three Arrenurus subgenera are 'natural': Arrenurus, Megaluracarus and Micruracarus, and the somewhat arbitrary distinction of the species assigned to the subgenus Truncaturus.

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.

Minimal barcode distance between two water mite species from Madeira Island: a cautionary tale.

In this work, we compare morphological and molecular data in their ability to distinguish between species of water mites (Acari, Prostigmata, Hydrachnidia). We have focused on the two species of the genus Lebertia inhabiting the island of Madeira. While traditional morphological traits were initially sufficient to distinguish between these two species, the molecular data were more dependable on the kind of analysis carried out. Single arbitrary genetic distance (e.g. a K2P distance below 2%) may lead to the conclusion that the specimens under study belong to the same species. Analysing the same specimens with the coalescent model has proved the evolutionary independence of both Lebertia clades in Madeira. Furthermore, multi-rate Poisson Tree Process analysis confirmed both lineages as independent species. Our results agree with previous studies warning of the dangers of rigid species delimitation based on arbitrary molecular distances. In addition, the importance of different molecular data approaches for correct species delimitation in water mites is highlighted.

Merging scleractinian genera: the overwhelming genetic similarity between solitary Desmophyllum and colonial Lophelia.

BACKGROUND: In recent years, several types of molecular markers and new microscale skeletal characters have shown potential as powerful tools for phylogenetic reconstructions and higher-level taxonomy of scleractinian corals. Nonetheless, discrimination of closely related taxa is still highly controversial in scleractinian coral research. Here we used newly sequenced complete mitochondrial genomes and 30 microsatellites to define the genetic divergence between two closely related azooxanthellate taxa of the family Caryophylliidae: solitary Desmophyllum dianthus and colonial Lophelia pertusa. RESULTS: In the mitochondrial control region, an astonishing 99.8 % of nucleotides between L. pertusa and D. dianthus were identical. Variability of the mitochondrial genomes of the two species is represented by only 12 non-synonymous out of 19 total nucleotide substitutions. Microsatellite sequence (37 loci) analysis of L. pertusa and D. dianthus showed genetic similarity is about 97 %. Our results also indicated that L. pertusa and D. dianthus show high skeletal plasticity in corallum shape and similarity in skeletal ontogeny, micromorphological (septal and wall granulations) and microstructural characters (arrangement of rapid accretion deposits, thickening deposits). CONCLUSIONS: Molecularly and morphologically, the solitary Desmophyllum and the dendroid Lophelia appear to be significantly more similar to each other than other unambiguous coral genera analysed to date. This consequently leads to ascribe both taxa under the generic name Desmophyllum (priority by date of publication). Findings of this study demonstrate that coloniality may not be a robust taxonomic character in scleractinian corals.

Development and characterization of 26 novel microsatellite loci for the trochid gastropod Gibbula divaricata (Linnaeus, 1758), using Illumina MiSeq next generation sequencing technology.

In the present study we used the high-throughput sequencing technology Illumina MiSeq to develop 26 polymorphic microsatellite loci for the marine snail Gibbula divaricata. Four to 32 alleles were detected per locus across 30 samples analyzed. Observed and expected heterozygosities ranged from 0.130 to 0.933 and from 0.294 to 0.956, respectively. No significant linkage disequilibrium existed. Seven loci deviated from Hardy-Weinberg equilibrium that could not totally be explained by the presence of null alleles. Sympatric distribution with other species of the genus Gibbula, as G. rarilineata and G. varia, lead us to test the cross utility of the developed markers in these two species, which could be useful to test common biogeographic patterns or potential hybridization phenomena, since morphological intermediate specimens were found.

Carinina ochracea (Palaeonemertea: Tubulanidae) Reaches Its Southernmost Distribution: New Morphological and Molecular Data.

New data for Carinina ochracea Sundberg et al., 2009 are provided for the Iberian Peninsula, establishing the southernmost limit of its known distribution. This species was previously known from only two localities: the type locality in Tjärnö (Sweden) and Pouldohan (Brittany, France). The material examined here was obtained during a faunal survey in the Villaviciosa Estuary (Asturias, northern Iberian Peninsula). The identity of the new specimen was confirmed both by DNA barcoding and anatomical examination. The molecular divergence of all available sequences of this species for four molecular markers, cytochrome c oxidase subunit I (COI), 16S, 18S and 28S rDNA, is discussed. For COI, four polymorphic sites were found, indicating: 1) a nuclear pseudogene; 2) heteroplasmy; or 3) gene duplication of a region of the mitochondrial genome. Two previously overlooked morphological characters were found: the presence of a colour ring and a postfixation staining band (pigmented band), which is histologically characterized. This species is the 12th palaeonemertean and the 75th nemertean reported from Iberian waters.

Development of Microsatellite Markers in the Deep-Sea Cup Coral Desmophyllum dianthus by 454 Sequencing and Cross-Species Amplifications in Scleractinia Order.

Microsatellite loci were isolated for the first time for the deep-sea coral Desmophyllum dianthus, using 454 GS-FLX Titanium pyrosequencing. We developed conditions for amplifying 24 markers in 10 multiplex reactions. Three to 16 alleles per locus were detected across 25 samples analyzed from Santa Maria di Leuca coral province (Mediterranean Sea). For the 24 polymorphic loci, observed and expected heterozygosities ranged from 0.211 to 0.880 and 0.383 to 0.910, respectively; 3 loci deviated from Hardy-Weinberg equilibrium, after null allele and sequential Holm-Bonferroni corrections. These newly isolated microsatellites are very useful genetic markers that provide data for future conservation strategies. Cross-amplification of these microsatellites, tested in 46 coral species, representing 40 genera, and 10 families of the phylum Cnidaria, produced informative allelic profiles for 1 to 24 loci. The utility of extending analyses to cross-species amplifications is also discussed.

Permanent genetic resources added to Molecular Ecology Resources Database 1 August 2012 - 30 September 2012.

This article documents the addition of 83 microsatellite marker loci and 96 pairs of single-nucleotide polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Bembidion lampros, Inimicus japonicus, Lymnaea stagnalis, Panopea abbreviata, Pentadesma butyracea, Sycoscapter hirticola and Thanatephorus cucumeris (anamorph: Rhizoctonia solani). These loci were cross-tested on the following species: Pentadesma grandifolia and Pentadesma reyndersii. This article also documents the addition of 96 sequencing primer pairs and 88 allele-specific primers or probes for Plutella xylostella.

Development of microsatellite markers as a molecular tool for conservation studies of the Mediterranean reef builder coral Cladocora caespitosa (Anthozoa, Scleractinia).

Cladocora caespitosa is a reef-building zooxanthellate scleractinian coral in the Mediterranean Sea. Mortality events have recurrently affected this species during the last decade. Thus, knowledge of its genetic structure, population diversity, and connectivity is needed to accomplish suitable conservation plans. In order to obtain a better understanding of the population genetics of this species, 13 highly variable microsatellites markers were developed from a naturally bleached colony. The developed primers failed to amplify zooxanthella DNA, isolated from C. caespitosa, verifying that these markers were of the coral and not algal symbiont origin. The degree of polymorphism of these loci was tested on tissue samples from 28 colonies. The allele number for each loci ranged from 2 to 13 (mean N(a) = 5.4), with an average observed heterozygosity of 0.42 (H(e) = 0.43) and all loci were in Hardy-Weinberg equilibrium. These new markers should be useful in future conservation genetic studies and will help to improve the resolution of the individual identification within this coral species. Primers were also tested in Oculina patagonica, with successful amplifications of several loci.

Leptospira interrogans: primeros aislamientos de humanos en Costa Rica / Lesptospira interrogans: first isolation of human in Costa Rica

La leptospirosis es una zoonosis que puede causar un amplio espectro de manifestaciones clínicas en humanos. Puede ser difícil orientar el diagnóstico, si no se toma en cuenta en el diagnóstico diferencial de los cuadros febriles agudos. Se requiere además, de técnicas especiales para obtener un diagnóstico microbiológico(1-4). Dado que en el Hospital de Ciudad Neily (HCN) son frecuentes los casos de pacientes con diagnóstico de cuadro febril agudo sin causa determinada y teniendo en cuenta el antecedente de una epidemia surgida en octubre de 1988 en Ciudad Cortés, donde el diagnóstico presuntivo de leptospirosis se estableció mediante datos clínicos y serológicos (5), nos propusimos aislar el agente etiológico para confirmar la existencia de infecciones por Leptospira interrogans en seres humanos, establecer el diagnóstico de algunas de estas fiebres de agudas y obtener serovares autóctonos para futuras investigaciones. En el período comprendido entre junio de 1995 y junio de 1996, se realizaron 228 hemocultivos por Leptospira sp en pacientes con cuadros febriles agudos, los cuales consultaron el Servicio de Emergencias Médicas del HCN; se obtuvieron 6 aislamientos (2,6 por ciento) identificados como Leptospira interrogans, tres de pacientes varones, dos de mujeres y uno de que por error en la identificación del cultivo no se pudo conocer su procedencia. Todos los aislamientos se obtuvieron entre los meses de julio a setiembre de 1995, coincidiendo con la época de mayor precipitación pluvial en la Región del Pacífico Sur del país. Se hace una breve descripción de los casos y finalmente se discute la importancia de obtener cepas autóctonas para la investigación y el desarrollo de métodos alternos de diagnóstico más oportunos, sensibles y específicos. Palabras claves: Leptospira interrogans-leptospirosis-aislamiento en humanos

Producción de penicilinasa (B-lactamasa en cepas de Neisseria gonorrhoeae, y sensibilidad in vitro, frente a algunos antibióticos / B-lactamase production in Neisseria gonorrhoea strains and in vitro antibiotic sensitivity

En un período de seis meses, se aisló 29 cepas de Neisseria gonorrhoeae, de las cuales un 28 por ciento fue B-lactamasaa positivo; el 72 por ciento fue B-lactamasa negativo, con un 8 por ciento de resistencia a la penicilina

Susceptibilidad a los antibióticos de los Staphylococcus sp coagulasa negativa aislada de muestras clínicas / Antimicrobial susceptibility of coagulase negative Staphylococcus sp from clinical samples

Devido a que Staphylococcus sp coagulasa negativa se considera como responsable de una gran variedad de infecciones, se estudió una muestra de 330 pacientes, de los cuales 39 (12%) fueron positivas por este microorganismo. Los porcentajes de resistencia general fueron penicilina 70, eritromicina 51, cloranfenicol 46, meticilina 41, gentamicina 0 y cefalosporina 0