Iván Acevedo

Two-way sex change in the endangered limpet Patella ferruginea (Mollusca, Gastropoda)

Direct observations of sex change were made on the endangered limpet Patella ferruginea in Chafarinas Islands (Alboran Sea) between 2006 and 2011. Individuals of the species were sexed and tagged during spawning season for subsequent monitoring to determine possible sex changes. Mortality was minimized by following a carefully designed sexing protocol. Out of 49 tagged specimens (41–88 mm in size) that could be sexed in successive years (28 males and 21 females), 16 males changed into females between consecutive years (50.0% of males smaller than 70 mm and 100% of males larger than this size), while two females changed sex to males between consecutive spawning seasons (both were smaller than 70 mm). Overall, 36.7% of the limpets monitored changed sex between consecutive years: 57.1% of males became females and 9.5% of females changed to males These observations confirms the occurrence of two-way sex change, or reverse sequential hermaphroditism, in P. ferruginea. Our findings unveil this sexual strategy in this endangered limpet and provide new direction for studies designed to address the mechanisms and factors that determine sex change and its effects on population dynamics.

Phylogeny of the reptilian Eimeria: are Choleoeimeria and Acroeimeria valid generic names?

Reptiles are the animals with the most described coccidian species among all vertebrates. However, the co‐evolutionary relationships in this host–parasite system have been scarcely studied. Paperna & Landsberg (South African Journal of Zoology, 24, 1989, 345) proposed the independent evolutionary origin of the Eimeria‐like species isolated from reptiles based on morphological and developmental characteristics of their oocysts. Accordingly, they suggested the reclassification of these parasites in two new genera, Choleoeimeria and Acroeimeria. The validity of the genera proposed to classify reptilian Eimeria species remained unresolved due to the lack of species genetically characterized. In this study, we included 18S rRNA gene sequences from seven Eimeria‐like species isolated from five different lizard host families. The phylogenetic analyses confirmed the independent evolutionary origin of the Eimeria‐like species infecting lizards. Within this group, most species were placed into two monophyletic clades. One of them included the species with ellipsoidal oocysts (i.e. Choleoeimeria‐like oocysts), whereas the species with more spheroidal oocysts (i.e. Acroeimeria‐like oocysts) were included in the second one. This result supports the taxonomic validity of the genera Acroeimeria and Choleoeimeria.

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.

Phylogenetic relationships of Isospora, Lankesterella, and Caryospora species (Apicomplexa: Eimeriidae) infecting lizards

In this study, several species of Isospora infecting lizards were genetically characterized. Specifically, five described and four newly described species of Isospora were included in a phylogeny of the family Eimeriidae. These species were isolated from hosts originally inhabiting all geographic continents except Europe. Phylogenetic analyses of the 18S ribosomal RNA (rRNA) gene grouped these nine species of Isospora with Lankesterella species and Caryospora ernsti. Therefore, within this clade, different evolutionary strategies in oocyst development and transmission occurred. Although the characteristic endogenous oocyst development of the genus Lankesterella may have arisen only once, the reduction in the number of sporocysts observed in the genus Caryospora occurred at least twice during coccidian evolution, as evidenced by the phylogenetic position of Caryospora bigenetica as the sister taxon of the group formed by reptilian Isospora, Lankesterella, and C. ernsti. Within this group, C. ernsti was the sister taxon to the genus Lankesterella. Overall, our results contradict the proposed monophyly of the genus Caryospora, highlighting the need for a thorough taxonomic and systematic revision of the group. Furthermore, they suggest that the recent ancestor of the genus Lankesterella may have been heteroxenous.

Development of tetranucleotide microsatellite markers for the cushion star, Asterina gibbosa, and cross-species amplification.

We describe nine polymorphic tetranucleotide microsatellite loci from the starfish, Asterina gibbosa. Loci were isolated from a partial genomic library that had been enriched for AAAC repeat sequences. Number of alleles per locus ranged from two to 14 in a sample of 85 individuals from three populations (two from Spain and one from the UK). Observed and expected heterozygosities per population ranged from 0.000 to 0.400 and from 0.040 to 0.784, respectively. All loci presented significant heterozygote deficits in one or more populations. Eight of these loci were amplified and variable in A. pancerii and A. phylactica. These loci will be used to study population structure in A. gibbosa.

Development and characterization of microsatellite markers for the endangered anchialine squat lobster Munidopsis polymorpha

Species of the genus Munidopsis are typically distributed in bathyal and abyssal zones, but the anchialine species Munidopsis polymorpha is an exception. It inhabits a volcanic tube on Lanzarote Island (Canary Islands, NE Atlantic) and is currently listed as endangered due to its highly restricted distribution and degree of endemism. Microsatellite loci were isolated from partial genomic libraries that had been enriched for AC, ACAG, GATA, AAAC and AAG repeat sequences. Eight loci were polymorphic in a sample of 24 individuals. The number of alleles per locus ranged from 2 to 4 with observed and expected heterozygosities ranging from 0.083 to 0.875 and from 0.080 to 0.681, respectively. These markers will be used to evaluate levels of genetic diversity and inbreeding, providing essential information for the development of a management and conservation strategy for this species.

Characterization of 17 novel microsatellite loci for the brown spiny lobster Panulirus echinatus Smith, 1869 using MiSeq

Using high-throughput sequencing technology 17 polymorphic microsatellites with perfect tetra-nucleotide repeats were identified for the brown spiny lobster Panulirus echinatus Smith, 1869. Two to ten alleles were detected per locus across 30 samples analysed from the Canary Archipelago. Observed and expected heterozygosities per locus ranged between 0.100 and 0.867 and 0.095 and 0.799, respectively. No significant linkage disequilibrium was found between pairs of loci, and only one locus (Pe-L43) deviated from Hardy-Weinberg equilibrium, likely due to the presence of null alleles. Despite its economic importance and the threat of overfishing, population genetics studies of this species are lacking. Therefore, these 17 novel microsatellites markers will be a useful genetic resource for future conservation studies of P. echinatus.

Looking for morphological evidence of cryptic species in Asterina Nardo, 1834 (Echinodermata: Asteroidea). The redescription of Asterina pancerii (Gasco, 1870) and the description of two new species

Abstract. Three species of the genus Asterina are known to inhabit the Mediterranean Sea and the north-eastern Atlantic Ocean: Asterina gibbosa (Pennant, 1777), A. pancerii (Gasco, 1870) and A. phylactica Emson & Crump, 1979. Differentiation of these species has primarily been based only on subtle characters (some highly debatable), such as colour or size. Therefore, this study aimed to review the morphological data characterising members of the genus, to incorporate new characters that may clarify morphological analyses and to couple morphological data with molecular evidence of differentiation based on the analysis of partial sequences of the cytochrome c oxidase subunit I (COI) and 18S rDNA (18S) genes and two anonymous nuclear loci (AgX2 and AgX5). The different lineages and cryptic species identified from the molecular analysis were then morphologically characterised, which was challenging given the limited number of diagnostic characters. Two of the five monophyletic lineages obtained molecularly (COI divergence >4%), further supported by differences in morphological characters and reproductive behaviour, are proposed as new species: Asterina martinbarriosi, sp. nov. from the Canary Islands, Spain (eastern central Atlantic Ocean) and Asterina vicentae, sp. nov. from Tarragona, north-eastern Spain (western Mediterranean Sea).

Development of microsatellite markers for the orange coral Astroides calycularis (Scleractinia, Dendrophylliidae). Permanent Genetic Resources added to Molecular Ecology Resources Database 1 August 2009–30 September 2009

This article documents the addition of 238 microsatellite marker loci and 72 pairs of Single Nucleotide Polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Adelges tsugae, Artemisia tridentata, Astroides calycularis, Azorella selago, Botryllus schlosseri, Botrylloides violaceus, Cardiocrinum cordatum var. glehnii, Campylopterus curvipennis, Colocasia esculenta, Cynomys ludovicianus, Cynomys leucurus, Cynomys gunnisoni, Epinephelus coioides, Eunicella singularis, Gammarus pulex, Homoeosoma nebulella, Hyla squirella, Lateolabrax japonicus, Mastomys erythroleucus, Pararge aegeria, Pardosa sierra, Phoenicopterus ruber ruber and Silene latifolia. These loci were cross‐tested on the following species: Adelges abietis, Adelges cooleyi, Adelges piceae, Pineus pini, Pineus strobi, Tubastrea micrantha, three other Tubastrea species, Botrylloides fuscus, Botrylloides simodensis, Campylopterus hemileucurus, Campylopterus rufus, Campylopterus largipennis, Campylopterus villaviscensio, Phaethornis longuemareus, Florisuga mellivora, Lampornis amethystinus, Amazilia cyanocephala, Archilochus colubris, Epinephelus lanceolatus, Epinephelus fuscoguttatus, Symbiodinium temperate‐A clade, Gammarus fossarum, Gammarus roeselii, Dikerogammarus villosus and Limnomysis benedeni. This article also documents the addition of 72 sequencing primer pairs and 52 allele specific primers for Neophocaena phocaenoides.