José E. F. Alfaya

Benthic diversity and assemblage structure of a north Patagonian rocky shore: a monitoring legacy of the NaGISA project

Fil: Rechimont, Maria Emilia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Nacional Patagonico; Argentina

Species Diversity of Ramphogordius sanguineus/Lineus ruber-Like Nemerteans (Nemertea: Heteronemertea) and Geographic Distribution of R. sanguineus.

Heteronemerteans, such as Lineus ruber, L. viridis, Ramphogordius sanguineus, R. lacteus, Riseriellus occultus, and Micrura varicolor, share many similar external characters. Although several internal characters useful for distinguishing these nemertean species have been documented, their identification is based mostly on coloration, the shape of the head, and how they contract, which may not be always reliable. We sequenced the mitochondrial COI gene for 160 specimens recently collected from 27 locations around the world (provisionally identified as the above species, according to external characters and contraction patterns, with most of them as R. sanguineus). Based on these specimens, together with sequences of 16 specimens from GenBank, we conducted a DNA-based species delimitation/identification by means of statistical parsimony and phylogenetic analyses. Our results show that the analyzed specimens may contain nine species, which can be separated by large genetic gaps; heteronemerteans with an external appearance similar to R. sanguineus/Lineus ruber/L. viridis have high species diversity in European waters from where eight species can be discriminated. Our 42 individuals from Vancouver Island (Canada) are revealed to be R. sanguineus, which supports an earlier argument that nemerteans reported as L. ruber or L. viridis from the Pacific Northwest may refer to this species. We report R. sanguineus from Chile, southern China, and the species is also distributed on the Atlantic coast of South America (Argentina). In addition, present analyses reveal the occurrence of L. viridis in Qingdao, which is the first record of the species from Chinese waters.

DNA barcoding supports identification of Malacobdella species (Nemertea: Hoplonemertea)

BackgroundNemerteans of the genus Malacobdella live inside of the mantle cavity of marine bivalves. The genus currently contains only six species, five of which are host-specific and usually found in a single host species, while the sixth species, M. grossa, has a wide host range and has been found in 27 different bivalve species to date. The main challenge of Malacobdella species identification resides in the similarity of the external morphology between species (terminal sucker, gut undulations number, anus position and gonad colouration), and thus, the illustrations provided in the original descriptions do not allow reliable identification. In this article, we analyse the relationships among three species of Malacobdella: M. arrokeana, M. japonica and M. grossa, adding new data for the M. grossa and reporting the first for M. japonica, analysing 658 base pairs of the mitochondrial cytochrome c oxidase subunit I gene (COI). Based on these analyses, we present and discuss the potential of DNA barcoding for Malacobdella species identification.ResultsSixty-four DNA barcoding fragments of the mitochondrial COI gene from three different Malacobdella species (M. arrokeana, M. japonica and M. grossa) are analysed (24 of them newly sequenced for this study, along with four outgroup specimens) and used to delineate species. Divergences, measured as uncorrected differences, between the three species were M. arrokeana-M. grossa 11.73%, M. arrokeana-M. japonica 10.62% and M. grossa-M. japonica 10.97%. The mean intraspecific divergence within the ingroup species showed a patent gap with respect to the interspecific ones: 0.18% for M. arrokeana, 0.13% for M. grossa and 0.02% for M. japonica (ranges from 0 to 0.91%).ConclusionsWe conclude that there is a clear correspondence between the molecular data and distinguishing morphological characters. Our results thus indicate that some morphological characters are useful for species identification and support the potential of DNA barcoding for species identification in a taxonomic group with subtle morphological external differences.

Development and characterization of microsatellite loci in the entocommensal Malacobdella arrokeana (Nemertea: Bdellonemertea), from Patagonia (Argentina) and cross-amplification in 34 nemertean species

Microsatellite loci (26 in total) were isolated for the first time for the entocommensal nemertean Malacobdella arrokeana, using 454 GS-FLX Titanium pyrosequencing. We developed conditions for amplifying these markers in 8 multiplex and 5 individual reactions. One to 14 alleles were detected per locus across 25 samples analyzed from San Matías Gulf, Patagonia (Argentina). For the 25 polymorphic loci, observed and expected heterozygosities ranged from 0.125 to 0.920 and 0.119 to 0.890, respectively; five loci deviated from Hardy–Weinberg equilibrium. Limited distribution and host specificity of M.arrokeana, which only inhabits Panopea abbreviata, have endangered this nemertean given the unregulated commercial exploitation of its host. These useful markers provide data for future conservation strategies. Cross-amplifications were also tested in 34 nemerteans species, representing the major lineages in this phylum. In total, 18 of the 26 loci (from one to 11 per species) gave clear allelic profiles.

Malacobdella arrokeana: Parasite or Commensal of the Giant Clam Panopea abbreviata?

We examined trophic relationship between the nemertean Malacobdella arrokeana and its host, the edible geoduck Panopea abbreviata by studying the diets of both species by direct (stomach contents) and indirect methods (stable-isotope analysis of C and N). In addition to these methods, the feeding behavior of M. arrokeana within the host and the morphology of its feeding organs were examined. The feeding behavior of M. arrokeana did not exhibit parasitic characteristics, and the proboscis morphology indicates it is unable to injure host tissues. Analysis of stomach contents revealed a diet consisting mainly of microalgae and diatoms. Panopea abbreviata and M. arrokeana shared similar trophic levels, presenting no differences in the spread of the isotopic niches and high overlap (SEAB overlapped 63%). Consistent with this, our results showed no differences in &dgr;15N or &dgr;13C values between the two species. The combination of direct and indirect approaches revealed that M. arrokeana has a diet similar to that of its host, confirming a commensal relationship.

Diet of the invasive sea slug Pleurobranchaea maculata in Patagonian coastal waters

ABSTRACT High densities of the invasive sea slug, Pleurobranchaea maculata, have been detected on the Argentinean coast, but studies on the species’ adaptation have been absent until now. Stomach contents of P. maculata in two sampling sites located at Golfo Nuevo, Patagonia Argentina, revealed animals as a major prey, but surprisingly algae ingestion on both sampling sites was abundant. This fact was not reported in previous studies that described opportunistic carnivorous feeding habits of species belonging to the genus Pleurobranchaea. Significant differences in the species prey items were found between sampling sites; however, sedentary polychaetes were the main consumed group at both sites. These results are useful to understand the adaptation of this invasive species in Argentinean waters and to monitor future changes in the benthic communities colonised by P. maculata.