Trace elements and vanadium in tissues and organs of five species of cetaceans from Italian coasts

Trace element concentrations (chromium, copper, zinc, iron, manganese and vanadium) were determined in organs of five species of cetaceans (Stenella coeruleoalba, Tursiops truncatus, Grampus griseus, Physeter macrocephalus, Ziphius cavirostris) that were found stranded along Italian coasts in the period 2000–2009. This dataset represents an important opportunity to verify and assess (particularly for V) patterns of incorporation of trace elements in different organs of cetaceans in a wide spectrum of species and related specimens distributed in all the age classes, and consequently determine the physiological and metabolic effects on the distribution modes of the same chemicals. In particular, Cu, Zn, and Fe accumulate preferentially in the liver of all studied specimens, while Mn and Cr values are found to be nearly constant in the analysed organs and tissues regardless of species. Comparable concentrations of trace elements, in different age classes, were measured for both specimens of S. coeruleoalba and T. truncatus (the most abundant dolphin species in the Mediterranean sea) in all analysed organs. On the other hand, unprecedented reported concentrations of V in tissues and organs of cetaceans from the Mediterranean show higher values when compared to levels measured in other marine mammals from the Atlantic Ocean.     

Sperm motility and longevity in the giant cuttlefish, Sepia apama (Mollusca: Cephalopoda)

The sperm kinetics and fertilisation literature in marine invertebrates is heavily biased toward free-spawning species. Nonetheless, many species (e.g. cephalopods) transfer and/or fertilise gametes in confined external spaces or internally, creating very different selective pressures on sperm storage, sperm longevity and hence sperm competition. Here we report the results of an investigation into the effects of sperm age, water temperature and sperm concentration on sperm motility in the giant cuttlefish (Sepia apama). Significant positive correlations were found between percent motility and sperm concentration, and between sperm motile speed and sperm concentration. Mean percent motility of cuttlefish sperm suspension was still 9% eight hours after being released from the spermatophore and diluted into filtered seawater at 12°C (ambient field temperature during the spawning season). Sperm resuspended from spermatangia taken from (mated) females in the field were motile for up to 100 hours. When spermatophores were stored at 4°C motility was still observed in resuspended sperm after two months. Our results show that spermatangia and spermatophores can retain and release live sperm for long periods. The observed longevity of sperm in S. apama greatly increases the potential for sperm competition in this species.     

Biomonitoring of polychlorinated byphenyls contamination in the supralittoral environment using the sandhopper Talitrus saltator (Montagu)

We evaluated the use of Talitrus saltator as biomonitor of polychlorinated byphenyls (PCBs) contamination of the supralittoral zone of Mediterranean sandy shores, an area not yet investigated about the presence of these pollutants. Twenty of 22 PCB congeners analysed were detected and quantified in sand and sandhoppers' samples collected in 6 different sites along the Tyrrhenian coast of Central Italy. Among the congeners analysed, PCB 153 and PCB 138 were the most abundant both in amphipods and sand samples and, regarding the congener pattern analysis, hexa-, hepta- and penta-PCBs were the most abundant congeners in sandhoppers. Though hydrophobic compounds, such as PCBs, bind strongly to sand and sediments, PCB concentrations were higher in T. saltator than in the sand revealing a good ability of this species to accumulate this class of contaminants. The concentration of each PCB congener recorded in tissues of sandhoppers showed significant differences among sampling sites revealing a good ability of these species to accumulate PCBs at different concentrations depending on the contamination levels of the sampling site. Therefore, our results suggest the possible utilisation of T. saltator as a biomonitor of PCB contamination of the supralittoral zone of Mediterranean sandy shores.     

From science to policy—a geostatistical approach to identifying potential areas for cetacean conservation: a case study of bottlenose dolphins in the Pelagos sanctuary (Mediterranean Sea)

Cetaceans are top-level predators that serve as sentinels of the health and status of lower trophic levels in the marine ecosystem. For this reason they attract significant attention in marine conservation planning and often have been used to promote designation of reserve areas in many countries (e.g., Ligurian Sea, Moray Firth, Hawaiian Islands, The Gully, Wadden Sea, Banks Peninsula, and Golfo San José). Many policies are designed to protect cetaceans. For example, the Habitat Directive requires member states to select, designate, and protect sites that support certain natural habitats or species, such as the bottlenose dolphin, as Special Areas of Conservation (SACs) that aim to create a network of protected areas across the European Union known as Natura 2000. The boundaries of protected areas for cetacean species must be defined for management purposes. In recent years, many techniques have been developed to define the distribution of cetaceans in relation to habitat preferences. Although these models can provide an understanding of the ecological processes that determine species distribution, their application requires prior knowledge of the variables that should be included in the model, the interactions among these variables, and their effects on species distribution. Thus, the lack of available data in understudied areas precludes the application of these types of models. As an alternative, we describe a geostatistical approach to identifying areas that potentially should be designated as marine protected areas for cetaceans. We illustrate the application of the kriging algorithm to the bottlenose dolphin population that resides in the northwestern Mediterranean Sea. The data derived from a 7-year survey were used. The encounter rate is the only variable required for this method, making it very easy to apply. The resulting georeferenced and high resolution map includes areas most visited by bottlenose dolphins, which are called core areas. Core areas are helpful for establishing the boundaries of marine reserves for the protection of the species. The approach described herein is accurate, precise, unbiased, replicable to all highly mobile species and easy to understand by both researchers and policy makers.     

International Study on Artemia. LVII. Morphological and molecular characters suggest conspecificity of all bisexual European and North African Artemia populations

A scanning electron microscopy (SEM) study of bisexual Artemia populations revealed that populations representing the species A. franciscana, A. persimilis, A. urmiana, A. sinica and a recently described species from Kazakhstan have a pair of spine-like outgrowths at the basal parts of their penes, whereas populations from southern Europe and North Africa (i.e. Mediterranean populations) lack these spine-like outgrowths. Allozyme and DNA polymorphisms, detected by allozyme starch gel electrophoresis and AFLP fingerprinting, respectively, suggested conspecificity of the studied populations from the broader Mediterranean basin. Male specimens from the collection of the Natural History Museum of London (UK) of the extinct A. salina population from Lymington lack spine-like outgrowths at the basal parts of the penes. This finding, based on a taxonomic character which is quite reliable, suggests conspecificity of A. salina from Lymington and the present bisexual Artemia populations from the Mediterranean basin, grouped under the binomen A. tunisiana. Received: 16 May 1997 / Accepted: 30 June 1997     

<Emphasis Type="Italic">Alexandrium catenella</Emphasis> (Dinophyceae), a toxic ribotype expanding in the NW Mediterranean Sea

The presence of the paralytic shellfish poisoning (PSP) dinoflagellate Alexandrium catenella in the north western (NW) Mediterranean Sea has been known since 1983. From this date on, the species has spread along the Spanish and Italian coastlines. Information concerning A. catenella isolates in the NW Mediterranean Sea was gained through phylogenetic studies. Twenty established toxic cultures of A. catenella taken from various NW Mediterranean Sea locations were analysed by nucleotide sequencing of the 5.8S rDNA and internal transcribed spacer regions. These rDNA ribosomal markers resulted useful in delineating the phylogenetic position of this species in the genus Alexandrium as well as in determining relationships between A. catenella isolates from different geographic areas. The phylogenetic position of the Mediterranean A. catenella ribotype, when compared to the “Alexandrium tamarense/catenella/fundyense species complex”, fits this species complex well. All the Mediterranean A. catenella isolates were constituted by only one genetic ribotype. By comparing the isolate sequences with those of other geographic areas, it revealed that the Mediterranean A. catenella ribotype was closely related to the A. catenella from Japan, Western Pacific Ocean. It was also evident that in temperate Japanese waters, a genetic variability was detected within A. catenella isolates; in fact, all strains resulted divergent showing as many as 15 mutational steps. The possibility that A. catenella has been recently introduced into the Mediterranean basin from temperate Asian areas is discussed.     

Patterns of courtship behavior and ejaculate characteristics in male red-winged blackbirds

Sperm competition in birds is likely to have important effects on the behavior and physiology of reproduction in both sexes. For males, such competition should select for large sperm reserves and behavioral adjustment of copulation when reserves are low. We investigated both these possibilities in free-living red-winged blackbirds (Agelaius phoeniceus), a highly polygynous species with apparently strong sperm competition. We found that the recent copulatory behavior of males did not affect the propensity to copulate with a model female. Ejaculates collected from individual males at 1-h intervals showed no evidence of sperm depletion, yet repeated ejaculates collected less than 10 min apart did. Male ejaculate size was significantly larger if it was the first one of the day (i.e., after an overnight rest). The average ejaculate size was 12.5 (±12.5 SD) million sperm. Males captured during the breeding season had an average of 111.7 (±52.8) million sperm stored in their seminal glomera. Because males average a peak copulation rate of six per female per day, in one day a male might utilize all the sperm in his seminal glomera if more than two females on his territory are fertilizable. We hypothesize that polygyny and sperm competition in this species have combined to select for rapid replenishment of the seminal glomera throughout the day, in contrast to other species that have been studied. Testis size and sperm reserves of male red-winged blackbirds are intermediate between monogamous species and species with intense sperm competition. Several possible explanations for this are discussed. Received: 21 October 1997 / Accepted after revision: 15 February 1998     

Trace metals in “ Posidonia oceanica ” seagrass from south-eastern Sicily

Heavy-metal concentrations were measured in sediments and tissues of Posidonia oceanica seagrass from south-eastern Sicily (Italy) in order to assess the degree of metal pollution in the coastal area. Seagrasses and sediments were collected at four sites along the south-eastern coast of Sicily. Flame atomic absorption spectrophotometry (FAAS) was used to measure concentrations of Cu, Zn, Cd and Pb. Standard statistical analyses were used to assess significant differences among the levels of the elements measured in different tissues and sediment and spatial distribution. The greatest values of potentially toxic metal concentrations were observed at the station near the industrial sites of Augusta and Priolo. Comparisons with the concentrations of the same metals in other sites of north and western Sicily and with data from surveys in other areas of the Mediterranean Sea are shown.     

Key Role of European Rabbits in the Conservation of the Western Mediterranean Basin Hotspot

Abstract: The Mediterranean Basin is a global hotspot of biodiversity. Hotspots are said to be experiencing a major loss of habitat, but an added risk could be the decline of some species having a special role in ecological relationships of the system. We reviewed the role of European rabbits (Oryctolagus cuniculus) as a keystone species in the Iberian Peninsula portion of the Mediterranean hotspot. Rabbits conspicuously alter plant species composition and vegetation structure through grazing and seed dispersal, which creates open areas and preserves plant species diversity. Moreover, rabbit latrines have a demonstrable effect on soil fertility and plant growth and provide new feeding resources for many invertebrate species. Rabbit burrows provide nest sites and shelter for vertebrates and invertebrates. In addition, rabbits serve as prey for a number of predators, including the critically endangered Iberian lynx (Lynx pardinus) and Spanish Imperial Eagle (Aquila adalberti). Thus, the Mediterranean ecosystem of the Iberian Peninsula should be termed “the rabbit's ecosystem.” To our knowledge, this is the first empirical support for existence of a multifunctional keystone species in a global hotspot of biodiversity. Rabbit populations have declined drastically on the Iberian Peninsula, with potential cascading effects and serious ecological and economic consequences. From this perspective, rabbit recovery is one of the biggest challenges for conservation of the Mediterranean Basin hotspot.     

Phylogeography of two species of <Emphasis Type="Italic">Lysidice</Emphasis> (Polychaeta,Eunicidae) associated to the seagrass <Emphasis Type="Italic">Posidonia oceanica</Emphasis> in the Mediterranean Sea

The aim of this work was to define the phylogeographic patterns of the two species of polychaete Eunicidae, Lysidice ninetta Audouin and Milne Edwards and Lysidice collaris Grube, both associated as sheath borers to the Mediterranean seagrass Posidonia oceanica, and with reference to their different origin and their actual geographic distribution. L. ninetta is distributed in the Atlantic and in the Mediterranean Sea while L. collaris is a tropical species, whose introduction into the Mediterranean Sea through the Suez Canal (lessepsian migrant) has been hypothesized in recent years. The two species have been often confused in the past, although they appear morphologically distinct. They share the same microhabitat (Posidonia sheaths) and they co-exist along a broad bathymetric range (1–30 m). Several populations for both taxa were sampled all along the coast of the Mediterranean basin. A variable no coding region of nuclear DNA (rDNA, ITS1) and a portion of a more conservative coding region of mitochondrial DNA (sub-unit one of citochrome oxidase, COI) were used as molecular markers. Both markers confirmed the separation between the two species. Low intraspecific polymorphism was present in L. collaris, together with absence of phylogeographic structure. In L. ninetta, instead, the presence of intraspecific cryptic lineages, sympatric in some sites, was recorded. Clustering of single populations in the two main clades was not always consistent between markers. The mitochondrial COI region showed more resolution at the given spatial scale. Our results suggest that Lysidice collaris could be recently introduced into the Mediterranean Sea from one or more separate events. On the other hand, for L. ninetta one could presume a re-colonization of the Mediterranean basin from the Atlantics, after the Messinian crisis (dry-out of the Mediterranean Sea, 5.5 my) with the subsequent separation of intraspecific lineages. The phylogeographic patterns of both Lysidice spp. are disjoined with respect to that of the host plant, P. oceanica. The obtained results suggest that environmental constraints and evolutionary history of these polychaetes and their host plant act in different ways to determine their actual genetic spatial structure.     

Niche segregation in sandy beach animals: an analysis with surface-active peracarid crustaceans on the Atlantic coast of Spain

Several species of sandy beach invertebrates regularly switch between burial in the sand during the day and surface activity at night to feed on stranded organic matter. Because all species consume essentially the same type of food (i.e., wrack) deposited over a restricted area, the potential for competition exists. Conversely, spatial and temporal segregation of surface activity behaviour is predicted to allow for niche separation. Here, we tested whether such behavioural niche separation occurs in three species of sympatric crustaceans (the talitrid amphipods Talitrus saltator (Montagu 1808) and Talorchestia brito Stebbing 1891, and the oniscoidean isopod Tylos europaeus Arcangeli 1938). We also assessed whether surface activity is modulated by weather, sea conditions and moon phases. Surface activity was measured over three consecutive moon phases on an exposed beach on the Atlantic Coast of Spain using pitfall traps along three transects from the foredunes to the swash. Adults of the amphipod T. saltator and the isopod T. europaeus overlapped spatially and temporally in their surface activity for most of the night. By contrast, the activity of T. brito was strongly disjunct in time and concentrated into significantly shorter bouts around dawn or dusk. Niche differentiation was also evident along the space axis, where T. brito occurred distinctly lower on the beach. Although several environmental factors (e.g., air and sand temperature, humidity, atmospheric pressure, wind speed and wind direction) were correlated with surface activity, this was highly variable amongst species, life stages, and synodic phases, and did not obscure the fundamental mosaic of spatio-temporal heterogeneity amongst species. Nocturnal feeding on beach wrack creates the potential for competition amongst sympatric crustaceans which can, however, be mitigated by behaviour patterns that separate consumers in time and space.     

Tropical fishes in a temperate sea: evolution of the wrasse <Emphasis Type="Italic">Thalassoma pavo</Emphasis> and the parrotfish <Emphasis Type="Italic">Sparisoma cretense</Emphasis> in the Mediterranean and the adjacent Macaronesian and Cape Verde Archipelagos

The northeastern Atlantic and the Mediterranean Sea share geological histories and display great faunal affinities. The majority of the Mediterranean species have Atlantic origins, with a few species with tropical affinities. These include the parrotfish Sparisoma cretense and the wrasse Thalassoma pavo that are restricted to the subtropical northeastern Atlantic, the Macaronesian archipelagos (Azores, Madeira, and Canaries) and the southern Mediterranean. The Pleistocene glaciations have been described as having different effects on the fauna of the two regions. During glacial peaks, Mediterranean waters remained warmer than those of the adjacent Atlantic. Within the eastern Atlantic, the effects of Pleistocene glaciations were differentiated. Here, we perform a comparative analysis focusing on T. pavo and S. cretense populations from the northeastern Atlantic and the Mediterranean to assess the effects of Pleistocene glaciations in these two species. Sequences from the mitochondrial control region were obtained and analyzed combining phylogeographic and demographic approaches. Gene flow between Atlantic and Mediterranean populations was shown to be very high. The Mediterranean populations of T. pavo and S. cretense showed high levels of genetic diversity, even in the eastern basin, pointing to an ancient colonization event. This suggests that both species must have been able to persist in the Mediterranean during the cold Pleistocene periods. Historical migration estimates revealed a Mediterranean towards Atlantic trend in the case of T. pavo, which may reflect the re-colonization of areas in the Atlantic by fish that survived the cold phases in relatively warmer Mediterranean refugia. Our data also showed that within the Macaronesian Archipelagos, migrations occurred from Madeira towards the Azores, for both T. pavo and S. cretense, thus supporting a post-glacial colonization of the Azores by fish that persisted in the warmer region of Madeira. Similar geographic distributions, thermal affinities, and means of dispersion for T. pavo and S. cretense resulted in a similar response to the effects of Pleistocene glaciations, as evidenced by identical phylogeographic patterns.     

Sperm quantity and quality effects on fertilization success in a highly promiscuous passerine, the tree swallow Tachycineta bicolor

Sperm competition is widespread among animal taxa and considered a major force in sperm evolution. Recent comparative studies have indicated that sperm competition selects for high sperm production capacity and long and fast-swimming spermatozoa across species. Here, we examine the role of sperm quantity and quality for fertilization success of individual males in a Canadian population of tree swallows Tachycineta bicolor, a socially monogamous, but highly promiscuous passerine. Male fertilization success (the sum of withinpair and extrapair young) was significantly associated with the size of the cloacal protuberance (a proxy for sperm quantity), but not with sperm size or in vitro sperm swimming speed. In a multivariate analysis, both cloacal protuberance volume and relative sperm midpiece size (i.e. high mitochondrial loading) had significant effects on male fertilization success. However, relative sperm midpiece size was not associated with fertilization success in a simple regression. Further, both cloacal protuberance volume and relative midpiece size had significant effects on sperm velocity, both in simple regressions and in a multivariate analysis. The finding that males with large relative midpiece size had both higher fertilization success and faster swimming sperm, suggests an indirect link between sperm morphology and male fertility mediated through sperm velocity. In conclusion, both quantitative and qualitative sperm traits seem to affect male fertilization success in tree swallows.     

Are mangroves and seagrasses sources of organic carbon for penaeid prawns in a tropical Australian estuary? A multiple stable-isotope study

The vertical distribution of the larvae of shelf-dwelling fish species that spawn in the NW Mediterranean Sea in spring was studied in relation to environmental data. Two sampling cycles were carried out at fixed stations on the continental shelf in May and June 1992. Three patterns of larval vertical distribution for the various taxa represented in the samples were observed. The larvae of most species (e.g. Boops boops, Diplodus sargus) were mainly located in the surface layer (10 m), others (e.g. Arnoglossus sp.) had broader distributions in the upper 40 m of the water column, and but a few (e.g. Gobiidae) were present in large concentrations at greater depths. The vertical distribution patterns of the various species showed no variations, despite high hydrographic variability during the study. The vertical distribution of only a few species (e.g. Arnoglossus sp., Crystallogobius linearis and Engraulis encrasicolus) varied over the diel cycle. The possible influence of the vertical distribution of fish larvae on their horizontal distribution patterns is discussed. Received: 10 March 1997 / Accepted: 4 April 1997     

Cephalopod remains from the stomach of a sperm whale stranded off Patagonia

The stomach of a male sperm whale (Physeter macrocephalus) stranded near Comodoro Rivadavia, Argentina, on 21 March 1986, contained 247 cephalopod beaks (128 upper and 119 lower). The lower beaks were identified, their rostral lengths were measured and used to estimate the mantle lengths and wet weight of the representatives of each cephalopod taxon. Fifteen species from nine families were represented. The most numerous species, in order of decreasing abundance, were the large cranchiidMesonychoteuthis hamiltoni (32.8% of total numbers),Mastigoteuthis sp. A (16.0%), andKondakovia longimana andHistioteuthis atlantica (both 13.5%). Almost 85% of the estimated total weight was contributed by two species,Mesonychoteuthis hamiltoni (61.6%) andK. longimana (23.1%). The presence of beaks fromK. longimana, Alluroteuthis antarcticus andM. hamiltoni, which are all thought to be restricted to the Antarctic, suggests that the whale had migrated North prior to stranding.     

Health risks of heavy metals in the mediterranean mussels as seafood

In recent years, mussels have already become commercially important seafood species worldwide. Mussels accumulate a wide range of metals in their soft tissue. Thus, the determination of accumulated concentrations of heavy metals, such as Pb, Cd, Hg, and As, in mussels is essential because of their usage as seafood and the potential adverse effects of their consumption on human health. In this review, these issues are presented and discussed using the Mediterranean mussel Mytilus galloprovincialis as an example. M. galloprovincialis is very efficient at converting low value victuals into high quality animal protein. The production of M. galloprovincialis in Mediterranean countries has been increasing rapidly, but Spain is still the largest producer of mussels. Only China has a larger production of these mussels than Spain. M. galloprovincialis is a filter feeding animal and accumulates a wide range of metals from their environment. The metal concentrations in the soft tissue of M. galloprovincialis are indicators of marine ecosystems contamination. In the same time, the bioaccumulation of heavy metals remains an issue concerning the consumption of mussels. Thereby, the Cd, Pb, Hg, and As concentration measurement in mussel soft tissue as a seafood have become significant. A review of literature data revealed large variations in the Cd, Pb, Hg, and As concentrations in M. galloprovincialis from their endemic areas, e.g., Mediterranean, Adriatic, and Black Sea, and the concentrations of these toxic metals were generally in the following order: As > Pb > Cd > Hg. The guidelines on heavy metals for seafood safety set by different countries and associations are reviewed. Comparison of the published data with European legislation showed that the levels of the heavy metals generally did not exceed the existing limits in all the mussels analyzed, excluding mussels from hot spots, such as lagoons and harbors, in the Mediterranean, Adriatic, and Black Sea.     

Sex in the beach: spermatophores, dermal insemination and 3D sperm ultrastructure of the aphallic mesopsammic Pontohedyle milaschewitchii (Acochlidia, Opisthobranchia, Gastropoda)

Sperm transfer via spermatophores is common among organisms living in mesopsammic environments, and is generally considered to be an evolutionary adaptation to reproductive constraints in this habitat. However, conclusions about adaptations and trends in insemination across all interstitial taxa cannot be certain as differences in mode of insemination via spermatophores do exist, details of insemination are lacking for many species, and evolutionary relationships in many cases are poorly known. Opisthobranch gastropods typically transfer sperm via reciprocal copulation, but many mesopsammic Acochlidia are aphallic and transfer sperm via spermatophores, supposedly combined with dermal fertilisation. The present study investigates structural and functional aspects of sperm transfer in the Mediterranean microhedylacean acochlid Pontohedyle milaschewitchii. We show that spermatophore attachment is imprecise. We describe the histology and ultrastructure of the two-layered spermatophore and discuss possible functions. Using DAPI staining of the (sperm-) nuclei, we document true dermal insemination in situ under the fluorescence microscope. Ultrastructural investigation and computer-based 3D reconstruction from TEM sections visualise the entire spermatozoon including the exceptionally elongate, screw-like keeled sperm nucleus. An acrosomal complex was not detected. From their special structure and behaviour we conclude that sperm penetrate epithelia, tissues and cells mechanically by drilling rather than lysis. Among opisthobranchs, dermal insemination is limited to mesopsammic acochlidian species. In this spatially limited environment, a rapid though imprecise and potentially harmful dermal insemination is discussed as a key evolutionary innovation that could have enabled the species diversification of microhedylacean acochlidians.     

Evolution of egg size and fertilisation efficiency in sea stars: large eggs are not fertilised more readily than small eggs in the genus<Emphasis Type="Italic"> Patiriella</Emphasis> (Echinodermata: Asteroidea)

Fertilisation kinetics theory suggests that, when sperm are limiting, the larger eggs of broadcast-spawning marine organisms ought to be fertilised more frequently than smaller eggs, because they provide a bigger target for searching sperm. Whilst this effect has been demonstrated within species, it is not known if this pattern holds among species. We tested whether a large difference in egg size between congeneric seastars with contrasting planktotrophic and lecithotrophic modes of development results in differences in the likelihood of eggs being fertilised in sperm-limiting situations. Measurement of egg sizes and sperm swimming speeds led to the prediction that the sperm–egg collision rate constant for Patiriella calcar (420-µm-diameter egg) should be nine times greater than for P. regularis (140-µm-diameter egg). Although the eggs of P. calcar should be fertilised at greater rates in low sperm concentrations, they were not. When gametes were allowed to mix for 10 s, the hypothesis that P. calcar eggs required less sperm than P. regularis to ensure 50% of eggs were fertilised was rejected. When gametes were mixed for 5 min, P. regularis eggs were more frequently fertilised, but the difference was not statistically significant. We conclude there must be a difference between these species in the likelihood that when a sperm finds a conspecific egg it can successfully fertilise. This apparent uncoupling of egg size and likelihood of fertilisation suggests that fertilisation is not a major constraint on the evolution of egg size in these seastars.Communicated by G.F. Humphrey, Sydney     

Low levels of genetic variation in mtDNA sequences over the western Mediterranean and Atlantic range of the sponge<Emphasis Type="Italic"> Crambe crambe</Emphasis> (Poecilosclerida)

Crambe crambe is a common encrusting sponge found in the Mediterranean and Atlantic littoral. An analysis of a partial sequence (535 bp) of the mitochondrial DNA (mtDNA) gene cytochrome oxidase subunit I (COI) was conducted in an attempt to determine population structure in this species. This is the first study of population genetics using this kind of marker in the phylum. Samples (N=86) were taken in eight populations separated by distances from 20 to 3,000 km, spanning from the western Mediterranean to the Atlantic. Low variability of this gene was found, as only two haplotypes were identified, along with low nucleotide diversity (=0.0006). However, the different frequencies found among populations revealed genetic structure and low gene flow between close populations, as expected from the dispersal biology of the species. The low variability found in sponges is in agreement with reports on cnidarians and points to a high conservation of mtDNA in diploblastic phyla.     

Spermatogenesis,sperm storage and comparative sperm morphology in nine species of Capitella,Capitomastus and Capitellides (Polychaeta: Capitellidae)

Light microscopy and transmission electron microscopy (TEM) were used to describe spermatogenesis and the morphology of mature sperm and sperm storage organs in five sibling species of Capitella, three species in the related genus Capitomastus, and one species in the genus Capitellides. These capitellids lack a well-developed testis, but young males have a few specialized regions of the peritoneum in the eighth setiger, where germ cells proliferate and spermatogonia are released into the coelom, and spermiogenesis is completed. Mature sperm are stored in the central regions of paired genital ducts (coelomoducts), which lie between the seventh and eighth setigers. The cells forming the walls of the coelomostome and central region of the duct are ciliated and have large glycogen deposits. The lumenal borders have extensive microvilli and there is evidence that they secrete glycogen-containing materials into the duct. All species have modified primitive sperm with a conical acrosome, elongated nucleus, and long middle piece extending along the proximal portion of the flagellum. A single ring-shaped mitochondrion encircles the centriolar region of the middle piece and the cytoplasm is filled with glycogen. The sperm of all nine species differ significantly in the lengths of their middle pieces, acrosomes and especially in their nuclear lengths. The nuclear lengths have a twofold range among the sibling species of Capitella and Capitomastus. Subtle differences in the shape and volume of the acrosomal vesicle and acrosomal space characteristic of the Capitella sibling species seem to correlate with a basic division of these species into those with diploid chromosome numbers of 20 or 26. Spermiogenesis, the number of sperm produced, and the method of sperm storage are appropriate for efficient sperm utilization in fertilization. No evidence indicates that spermatophores are formed and transferred between individuals and the method of sperm transfer is not understood. The differences in the dimensions and acrosome morphology of mature sperm, and the previously demonstrated specializations in the egg envelopes in the Capitella sibling species, are characteristic features of the reproductive isolation that exists among these capitellid species.