Highly polymorphic microsatellite markers for the Mediterranean endemic fan mussel Pinna nobilis

Pinna nobilis is an endemic bivalve of the Mediterranean Sea whose populations have decreased in the last decades due to human pressure; as a consequence, it was declared a protected species in 1992. Despite its conservation status, few genetic studies using mitochondrial markers have been published. We report on the isolation and development of 10 microsatellite loci for the fan mussel, Pinna nobilis. All loci (2 di-nucleotide, 5 tri-nucleotide, 2 tetra-nucleotide and 1 penta-nucleotide) are characterized by high levels of polymorphism in 76 individuals tested from two populations in the Balearic Islands (Spain, Western Mediterranean Sea). The number of alleles ranged from 4 to 24 and expected heterozygosity ranged from 0.4269 to 0.9400. These microsatellites could be very useful for the assessment of the genetic diversity and connectivity patterns of P. nobilis and the establishment of new conservation strategies.

Tasso di crescita di Pinna nobilis (Mollusca: Bivalvia) in due popolazioni, a Calpe e nell'Isola di Tabarca (Alicante, Mediterraneo Occidentale)

La conoscenza del tasso di crescita di due popolazioni di Pinna nobilis, in due località del Mar Mediterraneo Occidentale, insieme ad una stima d’età, sono un requisito fondamentale per poter comprendere meglio la biologia e l’ecologia di questa specie, al fine di poter istituire specifiche misure di tutela e conservazione. L’obiettivo di questa tesi è quello di utilizzare lo studio della sclerocronologia per comprendere l’origine dei registri interni di crescita e giungere così ad informazioni più accurate sulle modalità di accrescimento. Le valve di individui morti di Pinna nobilis sono state prelevate in due zone della costa Valenciana (Spagna), Calpe e Isola di Tabarca, al fine di valutare la variabilità del processo di crescita. Dalle valve sono state create delle sezioni di 300µm, grazie all’utilizzo di una sega con lama a diamante. Tali sezioni sono state osservate allo stereo-microscopio a luce trasmessa in direzione antero-posteriore. Una stima d’età iniziale è stata effettuata tramite una conta delle lingue di madreperla individuate. Per una stima più precisa è stato analizzato anche lo spessore dello strato di calcite in prossimità dei tre registri interni di crescita più vecchi. Gli individui più longevi appartengono alla popolazione dell’Isola di Tabarca (età max= 25anni), mentre l’età massima raggiunta dagli individui della popolazione di Calpe è 17 anni. Per analizzare l’accrescimento degli individui è stata utilizzata l’equazione di crescita di Von Bertalanffy. Gli individui della popolazione di Calpe hanno un tasso di accrescimento più elevato rispetto a quelli della popolazione di Tabarca, con una lunghezza massima asintotica (Lmax) di 65.2cm per Calpe e 54.9cm per l’Isola di Tabarca. Tali risultati confermano l’esistenza di una forte variabilità di accrescimento fra popolazioni di siti diversi. Le cause son da ricercare nelle caratteristiche dell’area, come il livello trofico, l’esposizione alla forza di onde e correnti o all’impatto antropico.

Analisi del tasso di crescita in Pinna nobilis (Molluschia Bivalvia): confronto fra una popolazione profonda e due superficiali del parco Nazionale di Cabrera (Isole Baleari, Spagna)

Il tasso di crescita di tre popolazioni di Pinna nobilis all’interno del Parco Nazionale di Cabrera (Isole Baleari) è stato analizzato per stimare l’età degli individui al fine di una migliore comprensione della biologia e dell’ecologia di questa specie sempre più minacciata. In questa tesi il tasso di crescita di una popolazione profonda, quella delle Baleari (20-30m), è stato confrontato con quello di due popolazioni superficiali: Freus e Gandulf (5-10m). L’obiettivo è valutare l’effetto della profondità e del sito sulle modalità di accrescimento degli individui. Sono state prelevate alcune valve di individui morti di P. nobilis a varie profondità. Lo studio della sclerocronologia ci ha permesso di studiare l’origine dei registri interni di crescita e le modalità di accrescimento. I risultati hanno mostrato una chiara differenziazione della taglia fra le tre popolazioni. Gli individui della popolazione profonda raggiungono dimensioni maggiori elevate e vivono più a lungo. Ma la popolazione di Gandulf ha un tasso di crescita simile a quello della popolazione delle Baleari essendo entrambe caratterizzate da individui di grandi dimensioni (Lmax Baleari= 65,1cm; Lmax Gandulf=62,4cm) e più longevi (età max=27anni). La popolazione di Freus è caratterizzata invece da individui di dimensioni minori e giovani di età (età max=14anni). Anche la velocità di crescita che porta a raggiungere la dimensione asintotica massima risulta simile nella popolazione di Gandulf (K=0,19) e nella popolazione delle Baleari (K=0,14), ed in entrambe è inferiore ai valori rilevati a Freus (K=0,21). Tali risultati evidenziano l’esistenza di una forte variabilità del tasso di accrescimento fra popolazioni collocate sia a profondità che in siti diversi. La differenza riscontrata tra la popolazione di Freus e quella di Gandulf è che la prima si trova in un sito esposto, mentre l’altra è collocata in una cala protetta subendo, quindi, un minor stress idrodinamico.

Using sclerochronology to estimate the growth rate in Pinna nobilis: a case study of Les Alfaques Bay population (Ebro Delta, Spain).

Pinna nobilis is the biggest Mediterranean bivalve, endemic and semi-infaunal. Provide hard substrates to colonize, increasing the spatial heterogeneity of the softbottom communities. P. nobilis suffer a drastic decline due to the anthropogenic pressures. It’s included in the Habitats Directive, in the Barcelona Convention, and in the red lists of many Mediterranean countries. Estimates the growth rate allows to understand the population dynamics of species and yield knowledge to improve protection efforts. In this study a new methodology based on sclerochronology was used to estimate the age and the growth rate of a P. nobilis population located in Les Alfaques bay. The shells of 35 specimens were cataloged. A subsample of 20 individuals was selected, and one valve of each specimens was cut into radial sections along PAMS (Posterior Adductor Muscle Scar) to study the inner register. Thus, the positions of PAMS obscured by nacre were identified, and the number of missing records was estimated by the width of the calcitic layer in the anterior part of the shell. The first growth curve for the Les Alfaques bay population was calculated from the length/age data. To simulate the growth rate of this population, the growth model based on the modified Von Bertalanffy equation was used. Shallow water usually hosts small sized populations of P. nobilis, while in deeper waters specimens reaches larger size. In Les Alfaques bay the population is composed by large size individuals though it’s located in shallows waters. This unusual size pattern is probably due to a sand bar that offers protection from hydrodynamic stress, allowing individuals to elongate more. This study contributes to the knowledge on P. nobilis biology and, with the aim to monitor this species, the growth curve could be used as baseline for future studies on habitat characteristics that may affect the population structure and dynamics in Les Alfaques Bay.

Studio sulla sclerocronologia e microstruttura delle valve malformate di Pinna nobilis per dimostrarne la capacità rigenerativa

Il presente studio si pone l’obbiettivo di indagare la capacità rigenerativa di P. nobilis, poiché sono stati osservati in natura individui con probabile ricostruzione delle valve a seguito di danneggiamenti, frequenti in questa specie sessile ed infossata solo in parte nel sedimento. La rigenerazione di parte del guscio potrebbe essere resa possibile in questa specie dalla presenza del mantello “fluido”, non fissato ai lobi della valva e di conseguenza notevolmente retrattile. Sono state analizzate le valve di 19 esemplari che presentavano anomalie strutturali. I campioni provengono da individui morti raccolti durante differenti campagne effettuate in tre aree mediterranee: delta dell’Ebro (Spagna), laguna veneta (Italia) e Port de Sète (Francia). L’analisi dei campioni è avvenuta attraverso l’osservazione microscopica delle sezioni dell’impronta del muscolo adduttore posteriore. In tre campioni, provenienti da aree diverse, è stato possibile documentare l’effettiva ricostruzione delle valve, resa evidente dall’intrusione di depositi di calcite all’interno della madreperla, che si inspessiscono gradualmente in direzione posteriore fino alla formazione della nuova conchiglia. Lo spessore sottile del nuovo strato di calcite e l’assenza delle marche annuali, caratteristiche simili della fase giovanile (in cui l’individuo si accresce velocemente), fa supporre che la ricostruzione avvenga molto rapidamente, ciò sarebbe giustificato dalla necessità di riparare velocemente il danno subito. In tre campioni, nonostante l’avvenuta divisione della valva, è assente il deposito di calcite all’interno della madreperla, probabilmente a causa della ricostruzione avvenuta ad un livello più posteriore. I restanti campioni hanno permesso di osservare la riparazione di piccole porzioni di guscio e dell’apice anteriore e la presenza di depositi di calcite più spessi, probabilmente dovuti al maggior apporto di proteine.

Allometric growth of a common Nassariidae (Gastropoda) in south-east Brazil

The allometric growth of two groups of Nassarius vibex on beds of the bivalve Mytella charruana on the northern coast of the State of Sao Paulo, was evaluated between September 2006 and February 2007 in the bed on Camaroeiro Beach, and from March 2007 to June 2007 at Cidade Beach. The shells from Camaroeiro were longer and wider and had a smaller shell aperture than those from Cidade; a principal components analysis also confirmed different morphometric patterns between the areas. The allometric growth of the two groups showed great variation in the development of individuals. The increase of shell width and height in relation to shell length did not differ between the two areas. Shell aperture showed a contrasting growth pattern, with individuals from Camaroeiro having smaller apertures. The methodology based on Kullback-Leibler information theory and the multi-model inference showed, for N. vibex, that the classic linear allometric growth was not the most suitable explanation for the observed morphometric relationships. The patterns of relative growth observed in the two groups of N. vibex may be a consequence of different growth and variation rates, which modifies the development of the individuals. Other factors such as food resource availability and environmental parameters, which might also differ between the two areas, should also be considered.

Database on the molluscan fauna from the Mediterranean Reef Ecosystems (CorMolDB - Part I)

In the Mediterranean Sea, infralittoral and circalittoral rocky bottoms (from 15 to 120 m) are characterized by a biogenic habitat, named "coralligenous", formed by the concretion of calcareous organisms, mainly algal thalli, and- to a lesser extent- by animal skeletons. This complex habitat is inhabited by a rich fauna that belongs to different taxonomic groups. Sponges, bryozoans, cnidarians and ascidians are the most common sessile organisms that inhabit the area while crustacean and molluscs are the common mobile organisms. Little information on the diversity of the molluscs that thrive in the coralligenous habitat is known while this information is highly important for biodiversity management purposes. After thoroughly studying the available and accessible published literature, a database for the molluscs of the coralligenous habitat has been designed and implemented for the collection and management of this information. From its index compilation more than 511 species of molluscs have been recorded so far from the coralligenous formations, the majority of which belongs to the class Gastropoda (357 sp.) followed by the Bivalvia (137 sp.), Polyplacophora (14 sp.), Cephalopoda (2 sp.) and Scaphopoda (1 sp.). Among these, the gastropod Luria lurida (Linnaeus, 1758) and Charonia lampas (Linnaeus, 1758), the endemic bivalve Pinna nobilis Linnaeus, 1758 and the endolithic bivalve Lithophaga lithophaga (Linnaeus, 1758), are protected by international conventions.

On the microstructure, growth pattern and original porosity of belemnite rostra: insights from calcitic Jurassic belemnites

Calcitic belemnite rostra are usually employed to perform paleoenvironmental studies based on geochemical data. However, several questions, such as their original porosity and microstructure, remain open, despite they are essential to make accurate interpretations based on geochemical analyses.This paper revisits and enlightens some of these questions. Petrographic data demonstrate that calcite crystals of the rostrum solidum of belemnites grow from spherulites that successively develop along the apical line, resulting in a “regular spherulithic prismatic” microstructure. Radially arranged calcite crystals emerge and diverge from the spherulites: towards the apex, crystals grow until a new spherulite is formed; towards the external walls of the rostrum, the crystals become progressively bigger and prismatic. Adjacent crystals slightly vary in their c-axis orientation, resulting in undulose extinction. Concentric growth layering develops at different scales and is superimposed and traversed by a radial pattern, which results in the micro-fibrous texture that is observed in the calcite crystals in the rostra.Petrographic data demonstrate that single calcite crystals in the rostra have a composite nature, which strongly suggests that the belemnite rostra were originally porous. Single crystals consistently comprise two distinct zones or sectors in optical continuity: 1) the inner zone is fluorescent, has relatively low optical relief under transmitted light (TL) microscopy, a dark-grey color under backscatter electron microscopy (BSEM), a commonly triangular shape, a “patchy” appearance and relatively high Mg and Na contents; 2) the outer sector is non-fluorescent, has relatively high optical relief under TL, a light-grey color under BSEM and low Mg and Na contents. The inner and fluorescent sectors are interpreted to have formed first as a product of biologically controlled mineralization during belemnite skeletal growth and the non-fluorescent outer sectors as overgrowths of the former, filling the intra- and inter-crystalline porosity. This question has important implications for making paleoenvironmental and/or paleoclimatic interpretations based on geochemical analyses of belemnite rostra.Finally, the petrographic features of composite calcite crystals in the rostra also suggest the non-classical crystallization of belemnite rostra, as previously suggested by other authors.

Comparative genetic study confirms exceptionally low genetic variation in the ancient and endangered relictual conifer, Wollemia nobilis (Araucariaceae)

The Wollemi pine, Wollemia nobilis (Araucariaceae), was discovered in 1994 as the only extant member of the genus, previously known only from the fossil record. With fewer than 100 trees known from an inaccessible canyon in southeastern Australia, it is one of the most endangered tree species in the world. We conducted a comparative population genetic survey at allozyme, amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) loci in W. nobilis, Araucaria cunninghatnii and Agathis robusta - representatives of the two sister genera. No polymorphism was detected at 13 allozyme loci, more than 800 AFLP loci or the 20 SSR loci screened in W. nobilis. In Ag. robusta only one of 12 allozyme loci, five of 800 AFLP loci and none of the 15 SSR loci were variable. For A. cunninghamii, 10 of > 800 AFLP loci and five of 20 SSR loci were variable. Thus low genetic diversity characterizes all three species. While not ruling out the existence of genetic variation, we conclude that genetic diversity is exceptionally low in the Wollemi pine. To our knowledge this is the most extreme case known in plants. We conclude that the combination of small population effects, clonality and below-average genetic variation in the family are probable contributing factors to the low diversity. The exceptionally low genetic diversity of the Wollemi pine, combined with its known susceptibility to exotic fungal pathogens, reinforces current management policies of strict control of access to the pines and secrecy of the pine locations.

The fan mussel Pinna rudis : occurrence and association with Pontonia pinnophylax in Terceira Island (Azores, NE Atlantic).

Conference: IMMR | International Meeting on Marine Research 2014. Peniche, Portugal, 10 Jul - 11 Jul, 2014. Presentation Type: Poster Presentation.

Effects of Laurus nobilis (Lauraceae) on Biomphalaria glabrata (Say, 1818)

Experiments were carried out using aqueous extracts from leaves and flowers of Laurus nobilis on Biomphalaria glabrata. Treatments were performed on blastula stage (± 15 h after first cleavage) and on adult snails (11-18 mm). In both instances they were exposed for 24 h to different concentrations of the extracts on snails (200 to 2500 ppm) and embryos (20 to 300 ppm) at 25 ± 1ºC. The embryos were observed for a period of 20 days after treatment and the snails for 10 days. Results obtained with leaf aqueous extracts have shown a degree of toxicity on embryos starting at a concentration of 125 ppm, the flower extract being effective at 35 ppm. The malformation obtained with the different concentrations falls into the unespecific type category, however some cephalic and shell malformations were found in embryos treated with concentrations over 50 ppm (leaves) and 25 ppm (flowers). The LD90 on adult snails obtained by treatments with flower and leaf extract was observed at concentrations of 340 ppm and 1900 ppm respectively.

Seguimiento de la Pradera de Posidonia oceanica de las Islas Medas. Ejercicio de 1999

Siguiendo la metodología de años anteriores(ver informes de seguimiento previos), durantelos meses de octubre a diciembre de 1999 setomaron una serie de medidas de vitalidad de lapradera de Posidonia oceanica de las islas Medes,así como de macrofauna asociada.Los resultados se presentan en las tablas quese adjuntan.No se han detectado cambios notables en losdescriptores de vitalidad de la planta (densidad ycobertura).No se han detectado inflorescencias.La población de erizos de la pradera se mantieneestable.Las nacras (Pinna cf. nobilis) han experimentadoun crecimiento significativo en anchura y enaltura, no habiéndose detectado ninguna mortalidaden la población estudiada en 1999. Se hanidentificado en el presente ejercicio tres individuosnuevos, de los que uno corresponde probablementea un recluta.Del seguimiento detallado del balizamientodel calvero se desprende que no hay cambiosimportantes en los límites de esta estructura

Frugivory on Margaritaria nobilis L.f. (Euphorbiaceae): poor investment and mimetism

Dehiscent fruits of Euphorbiaceae usually have two stages of seed dispersal, autochory followed by myrmecochory. Two stages of Margaritaria nobilis seed dispersal were described, the first stage autochoric followed by ornithocoric. Their dehiscent fruits are green and after they detached from the tree crown and fall on the ground, they open and expose blue metallic cocas. We studied the seed dispersal system of Margaritaria nobilis in a semi-deciduous forest in Brazil. In 80 h of focal observations, we recorded only 12 visits of frugivores, however the thrush Turdus leucomelas was the only frugivore that swallowed the fruits on the tree crown. Pitylus fuliginosus (Fringilidae) and Pionus maximiliani (Psittacidae) were mainly pulp eaters, dropping the seeds below the tree. On the forest floor, after fruits dehiscence, jays (Cyanocorax chrysops), guans (Penelope superciliaris), doves (Geotrygon montana) and collared-peccaries (Pecari tajacu) were observed eating the blue diaspores of M. nobilis. Experiments in captivity showed that scaly-headed parrots (Pionus maximiliani), toco toucans (Ramphastos toco), jays (Cyanochorax chrysops), and guans (Penelope superciliaris) consumed the fruits and did not prey on the seeds before consumption. The seeds collected from the feces did not germinate in spite of the high viability. The two stages of seed dispersal in M. nobilis resembles the dispersal strategies of some mimetic species. However M. nobilis seeds are associated with an endocarp, it showed low investment in nutrients, and consistent with this hypothesis, M. nobilis shared important characteristics with mimetic fruits, such as bright color display, long seed dormancy and protection by secondary compounds.