Variation in fertilisation abilities between hemiclonal hybrid and sexual parental males of sympatric water frogs (<Emphasis Type="Italic">Rana lessonae</Emphasis>,<Emphasis Type="Italic"> R. esculenta</Emphasis>,<Emphasis Type="Italic"> R. ridibunda</Emphasis>)

In many species, males and females mate with multiple partners, which gives rise to sperm competition and multiple paternity. The experiments on water frogs presented here demonstrate that such sperm competition can affect the structure and dynamics of mixed-species communities. The hybrid frog Rana esculenta (LR) mates with one of its parental species, usually R. lessonae (LL), although in some areas R. ridibunda (RR), to regain the premeiotically eliminated parental genome ("hybridogenesis"). Mixed LL/LR-populations are stable although hybrid numbers should continuously increase at the expense of parental animals, because of differences in female fecundity and other factors. This would finally lead to the extinction of the sexual host, followed by that of the sexual parasite, unless the reproductive superiority of R. esculenta is reduced by other factors, such as lower hybrid male fertility. Eggs from LL- and LR-females were fertilised in vitro by single- and multi-male sperm suspensions of LL-, LR- and RR-males. In all experiments, the proportion of offspring sired by R. esculenta sperm was significantly lower than that sired by R. lessonae or R. ridibunda sperm. Gonad mass, sperm morphology, sperm swimming velocity, and sperm survival did not explain these differences in fertilisation success; nor did gamete recognition and compatibility. Sperm density was the only trait that paralleled fertilisation success, but it offers no explanation either, because densities were equalised for the in-vitro fertilisations. In natural LL/LR populations, the significantly smaller amount, poorer competitive ability and lower long-term survival of R. esculenta compared to R. lessonae sperm will reduce the initial reproductive superiority of hybrids and contribute to the stabilisation of mixed water-frog populations. Differences in fertilisation ability are also likely to be relevant for the structure and dynamics of several other systems with encounters between eggs and sperm from different genotypes, ecotypes, ploidy levels and/or species.     

Ecology of a corallivorous gastropod,<Emphasis Type="Italic"> Coralliophila abbreviata</Emphasis>, on two scleractinian hosts. I: Population structure of snails and corals

Despite their potential importance in structuring reef communities, invertebrate corallivores and their population structures are poorly understood. We found distinct differences in the population structures (length-frequency distribution and sex ratio) of the corallivorous gastropod Coralliophila abbreviata residing on two coral-host taxa, Montastraea spp. and Acropora palmata, in the Florida Keys. In each of two survey years, around 50% of the Montastraea spp. colonies were infested, with a mean snail density of eight snails per infested colony (range 1–45), while around 20% of A. palmata colonies harbored three snails per infested colony (range 1–23). Variation in patterns of snail occurrence was also observed within a host taxon. A. palmata occurred in low- and high-density stands (0.4 and 1 colony m^–2, respectively, at the initial survey) at different sites. Hurricane Georges struck the area in September 1998. When resurveyed in 1999, density of colonies in low-density stands had decreased by 75% to 0.1 colonies m^–2. This decrease was accompanied by a doubling in the proportion of colonies infested with snails (from 19% to 46%) and an increase in snail density per infested colony (from 3.7±3.3 SD to 5.4±4.6 SD) as snails apparently concentrated on surviving A. palmata. In contrast, sites with high density A. palmata stands (thickets) retained colony densities of about ~1 colony m^–2 among years, while snail infestation increased only from 9% to 14% of colonies surveyed and snail density essentially remained unchanged (from 2.7±1.8 to 2.9±1.9 snails per infested colony). Snails collected from Montastraea spp. were shorter than those from A. palmata in low-density stands and were longest on A. palmata in thickets. On both host taxa, female snails were longer than males. The sex ratio of snails on Montastraea spp. hosts was even (1:1), while that of snails on A. palmata was skewed (70% males). Factors that could explain observed differences in size structure and sex ratio between Coralliophila populations on the two coral host taxa include: differential susceptibility to predators, influence of host tissue nutritional quality and/or secondary metabolite content, and genetic differences (cryptic species). The host-specific characteristics of C. abbreviata populations imply that the impact of gastropods on reef communities will vary with the coral species composition.     

Genetic structure of<Emphasis Type="Italic"> Linckia laevigata</Emphasis> and<Emphasis Type="Italic"> Tridacna crocea</Emphasis> populations in the Palawan shelf and shoal reefs

Allozyme variation of 10 populations of Linckia laevigata at 8 polymorphic loci and 13 populations of Tridacna crocea at 6 polymorphic loci were analyzed to compare genetic variability and genetic affinities among reefs in Palawan, Philippines. Two to five populations were sampled from each of four regions: the shelf reefs in (1) northern Palawan and (2) southern Palawan and the offshore reefs in (3) the Kalayaan island group (KIG) in the South China Sea and (4) the Tubbataha shoals in the Sulu Sea. Heterozygosity was highest in populations of L. laevigata from the south shelf of Palawan and populations of T. crocea from the Tubbataha shoals of the Sulu Sea. The lowest heterozygosity estimates were from the reefs of the KIG in the South China Sea, for both species. Overall F _ST values for both species were significant, with an estimated average number of effective migrants per generation (N _EM) of 4.85 (~5 individuals) for L. laevigata and 3.54 (~4 individuals) for T. crocea. Within-region comparisons showed N_EM ranging from 6.29 to 92.34 for L. laevigata and from 3.40 to 6.30 for T. crocea. The higher gene flow among L. laevigata populations relative to T. crocea is consistent with the greater dispersal potential of the former species. Finer scale genetic structuring was evident in T. crocea populations. For both species, the Tubbataha reefs in the Sulu Sea have higher genetic affinity with the populations from the southern shelf of Palawan, while the reefs in the KIG had higher affinity with the northern Palawan shelf reefs. The north and south shelf populations have the least genetic affinity. Genetic patchiness among reefs within regions suggests the importance of small-scale physical factors that affect recruitment success in structuring populations in small island and shoal reef systems in Palawan.     

Physiological adaptations of the invasive cordgrass<Emphasis Type="Italic"> Spartina anglica</Emphasis> to reducing sediments: rhizome metabolic gas fluxes and enhanced O<Subscript>2</Subscript> and H<Subscript>2</Subscript>S transport

Cordgrasses of the genus Spartina form dense monospecific stands worldwide, profoundly influencing the ecology of estuaries. One species, Spartina anglica, originated by allopolyploidy in the 1800s and has been particularly prolific as an invasive species worldwide. S. anglica tolerates low-lying estuarine mudflats that its progenitor species and other coastal halophytes cannot. However, very little is known of the physiology of S. anglica. In the present study, an automated flow-through respirometry system was used to quantify metabolic gas fluxes (O₂, H₂S, CO₂, and NH₃) of S. anglica rhizomes. Enhanced physiological mechanisms to transport O₂ and H₂S in both directions between the rhizosphere and the atmosphere were exhibited by S. anglica, but not by the native North American species S. alterniflora. These results suggest that tolerance of anoxia and H₂S may assist S. anglica in colonizing extensively flooded environments. Enhanced sediment oxygenation by S. anglica may be potentially useful for phytoremediation of contaminated sediments, since microbial degradation of organic pollutants is often limited by O₂ availability.     

Population structure of two northern hemisphere polychaetes,<Emphasis Type="Italic"> Neanthes virens</Emphasis> and<Emphasis Type="Italic"> Hediste diversicolor</Emphasis> (Nereididae), with different life-history traits

The present study compared population genetic structure of two northern hemisphere polychaetes, Neanthes virens (Sars, 1835) and Hediste diversicolor (O.F. Müller, 1776), with different life-history traits. N. virens has a pelagic larval phase that may last as long as 9 days, while H. diversicolor broods its offspring. It was predicted that N. virens populations, especially over relatively short geographic distances, would be more genetically similar than H. diversicolor populations. Sequence data were obtained for two mitochondrial genes, cytochrome c oxidase I (COI) and cytochrome b (cyt b). The COI gene showed no variation in 16 N. virens individuals from 12 locations, and it was not examined in H. diversicolor. The nucleotide sequence of the cyt b gene was examined in 93 N. virens individuals collected in 1992, 1999, and 2000 from 12 localities throughout its range (North America, Europe, Japan) and from 33 H. diversicolor individuals collected in 1999 and 2001 from one locality in eastern Canada and two localities in France (Roscoff and Marseille). In N. virens, one cyt b haplotype was shared by 89% (83 out of 93) and only two other haplotypes were found. Significant population structure was observed in H. diversicolor. In the 33 sequences compared, 18.9% of sites were polymorphic and 12 haplotypes were identified. There were two haplotypes in the Canadian population, three in the Marseille samples, and seven in the Roscoff samples, and no haplotype was found in common between any two sites. These results show that widely separated H. diversicolor populations are strongly differentiated at the cyt b locus. The most plausible interpretation for the low variation at the COI and cyt b loci in N. virens, and for the lack of variation at allozyme loci (based on an initial survey of a smaller subset of populations), is that this species has experienced a drastic bottleneck and that a few remaining lineages recolonized its entire range.     

Comparing biochemical changes and energetic costs in gastropods with different developmental modes: <Emphasis Type="Italic">Crepipatella dilatata</Emphasis> and <Emphasis Type="Italic">C. fecunda</Emphasis>

The Chilean gastropods Crepipatella dilatata and C. fecunda have different development modes: brooding and direct development in C. dilatata and brooding and planktotrophic development in C. fecunda. Unlike many other congeneric invertebrate species pairs, recent genetic evidence suggests that C. fecunda may have evolved from C. dilatata. To explore the changes involved in this unusual evolutionary path, this study examined the biochemical, energetic, and morphological characters during early development of both species. Mean egg size was slightly smaller for the direct-developing species C. dilatata, and initial energy content was lower—by about 27%—for eggs of that species. In both species, protein content in the eggs was the principal biochemical component. Although females of C. fecunda produce 180 times more eggs than C. dilatata, females of C. dilatata invest 20 times more energy in each of their offspring, through nurse eggs; their embryos have approximately eight times more energy at hatching and about 5 times more energy when they enter the benthos, despite a long planktonic feeding period in the larvae of C. fecunda. Evolutionary switching between modes of development in these species is reflected in shifts in maternal energy investment.     

Evolution of coral reef fish<Emphasis Type="Italic"> Thalassoma</Emphasis> spp. (Labridae). 1. Molecular phylogeny and biogeography

Wrasses in the genus Thalassoma comprise 27 recognized species that occur predominantly on coral reefs and subtropical rocky reefs worldwide. The phylogenetic relationships for 26 species were examined based on two mitochondrial genes (cytochrome b and 16S rRNA) and one nuclear intron (the first intron of the ribosomal protein S7). Two closely related species, the bird-wrasses (Gomphosus varius Lacepède, 1801 and G. caerulaeus Lacepède, 1801), were also included in the analysis. These species grouped within the genus Thalassoma. Thalassoma newtoni (Osório, 1891) from Sao Tome, which is generally synonymized with the Atlantic/Mediterranean Thalassoma pavo (Linnaeus, 1758) appears to be a valid species. Using a molecular clock, the genus was estimated to have originally diverged 8–13 million years ago, with Thalassoma ballieui (Vaillant and Sauvage, 1875) from Hawaii and Thalassoma septemfasciata Scott, 1959 from Western Australia as the ancestral species. Approximately 5–10 million years ago, a sudden burst of speciation resulted in seven clades, which were resolved with the sequence data. The terminal Tethyan event and the closing of the Isthmus of Panama were probably the major historical factors shaping the evolution of species in the genus Thalassoma. These data on the spatio-temporal pattern of speciation in the Indo-Pacific indicate that peripheral species have been generated at various times throughout the history of the genus, and that none of the widespread species are relatively young. Thus, there is no clear support for centrifugal (youngest at the periphery) versus centripetal (oldest at the periphery) modes of generation of species, two theories which have been used to account for geographic gradients in species diversity.     

Differences in the species- and size-composition of fish assemblages in three distinct seagrass habitats with differing plant and meadow structure

Fish faunas were sampled seasonally using a large and a small beam trawl in three seagrass habitats comprising predominantly Amphibolis griffithii or Posidonia sinuosa or Posidonia coriacea, which differ in seagrass and meadow structure. Amphibolis griffithii and P. sinuosa both produce a relatively dense leaf canopy, but the former exhibits a distinct architecture, with the leaf canopy overlying relatively open spaces surrounding woody stems, compared to the uniformly dense blade-like leaves of P. sinuosa which emerge directly from the sediment. In comparison, P. coriacea provides a landscape of patchy seagrass amongst areas of bare sand. Since the latter seagrass habitat contains large areas of sand, fish were also sampled in adjacent unvegetated areas. The number of species and density of fish were greater (P<0.05) in P. sinuosa than in either A. griffithii or P. coriacea. The mean number of species caught using the large trawl ranged from 16 to 24 in the first of these habitats compared to 14–21 and 9–15 in the last two habitats, respectively, and the mean densities ranged between 78 and 291 fish 1000 m^?2 in P. sinuosa compared to 31–59 fish 1000 m^?2 in A. griffithii and 31–59 fish 1000 m^?2 in P. coriacea. The biomass of fish was greater (P<0.05) in both P. sinuosa and A. griffithii than in P. coriacea (4.2–5.3 kg and 3.3–6.2 kg versus 0.7–1.9 kg 1000 m^?2, respectively). Furthermore, the size-structure of fish differed among these habitats, where the median weight of fish was 72.1 g in A. griffithii, compared to 7.5 g and 19.8 g in P. sinuosa and P. coriacea, respectively. Ordination and ANOSIM demonstrated that the species-composition differed markedly among the three seagrass habitats (P<0.001), suggesting that fish species display a distinct preference for particular seagrasses characterised by different architecture. Differences in species-composition among the seagrass habitats partly reflected the size-composition of fish in each habitat. The open space below the canopy of A. griffthii is likely to allow larger fish to occupy this habitat, whereas only small fish would be able to penetrate the dense foliage of P. sinuosa. Differences in species- and size-composition of fish among these habitats may be the result of settlement-sized larvae discriminating between particular seagrass and meadow structures, or fish being subject to different levels of predation and/or accessibility to food or space. The species-composition in P. coriacea was highly dispersed and did not differ from that of unvegetated areas. While several species were associated with both P. coriacea and bare-sand habitats, some species did display a high affinity with the seagrass P. coriacea. This may reflect an association with the sparse and narrower leaves of this seagrass or with the patchy occurrence of the seagrass Heterozostera tasmanica, which commonly occurs as an understorey in this habitat.     

Real-time PCR assay for detection and relative quantification of <Emphasis Type="Italic">Liocarcinus </Emphasis><Emphasis Type="Italic">depurator</Emphasis> larvae from plankton samples

Accurate species identification of decapod crustacean larvae is required to understand their population distributions, life cycle dynamics and interactions with their habitats. Analysis of plankton samples using morphological taxonomic methods and microscopy is time-consuming, requires highly skilled and trained operatives and may often be inaccurate. As complementary tools to classical identification methods, recent work has focused on the development of molecular approaches and shows their feasibility for species-specific identification. This study has developed real-time PCR assays utilising species-specific Taqman^® probes designed in the cytochrome oxidase I (COI) gene of Liocarcinus depurator, Necora puber, Carcinus maenas and Cancer pagurus. Our study then employed the probe and primers designed for L. depurator to obtain accurate identification and relative abundance estimates of L. depurator larvae in plankton samples collected between March 2005 and October 2006. Ranges of larval abundances were derived from a standard curve created from plankton samples spiked with a known number of larvae reared in the laboratory. Inhibition of the PCR reaction was shown to be an important factor and our results suggested that 0.1 ng of DNA as template provided accurate identification and avoided inhibition. Real-time PCR was shown to provide accurate species identification on unsorted plankton samples and could be suitable for the estimation of larval abundances in the plankton, although more work must be done to improve the accuracy of those estimations.     

Patterns of gamete incompatibility between the blue mussels<Emphasis Type="Italic"> Mytilus edulis</Emphasis> and<Emphasis Type="Italic"> M</Emphasis>.<Emphasis Type="Italic"> trossulus</Emphasis>

Previous research on gametic incompatibility in marine invertebrates suggests that for highly dispersive marine invertebrate species, barriers to fertilization among closely related taxa are often incomplete and sometimes asymmetric. The nature of these barriers can dramatically affect the patterns of gene flow and genetic differentiation between species, and thus speciation. Blue mussels, in the genus Mytilus, are genetically distinct in allopatry yet hybrids are present wherever any two species within the group co-occur. The present study sampled M. edulis (L.) and M. trossulus (Gould) in May and June 2001 from the East Bay section of Cobscook Bay, Maine, USA (latitude 44°56′30″N; longitude 67°07′50″W), where the two species are sympatric. Gamete incompatibility was investigated in a series of laboratory fertilizations carried out in July 2001. The proportion of fertilized eggs typically exceeded 80% at sperm concentrations of 10³–10⁴ ml^?1 among intraspecific matings (n=18), but was <30% even at sperm concentrations in excess of 10⁵–10⁶ ml^?1 for interspecific matings (n=13). Further analysis indicated that approximately 100- to 700-fold higher sperm concentrations were required to achieve 20% fertilization in interspecific matings relative to intraspecific matings, indicating strong barriers to interspecific fertilization. The proportion of fertilized eggs did not follow this general pattern in all matings, however. The eggs from two (out of five) M. edulis females were almost as receptive to M. trossulus sperm as they were to M. edulis sperm. In contrast, the eggs from all M. trossulus females (n=3) were unreceptive to M. edulis sperm, suggesting that fertilization barriers between these species may be asymmetric. Given the experimental design employed in this study, the results are also consistent with a strong maternal or egg effect on the level of interspecific gamete compatibility in M. edulis.     

Blood metabolites and hemocyanin of the white shrimp,<Emphasis Type="Italic"> Litopenaeus vannamei</Emphasis>: the effect of culture conditions and a comparison with other crustacean species

Oxyhemocyanin (OxyHc), blood protein, glucose, acylglycerol (AG), and lactate of Litopenaeus vannamei juveniles were investigated. Statistical analyses were done on baseline values for each metabolite and on OxyHc from shrimp cultured in outdoor (20-m² ponds) and indoor (90-l tanks). The values obtained were compared with blood metabolites reported in the literature for L. vannamei maintained under similar conditions. Normal distribution was only found for samples from indoor shrimp. The distribution values for blood metabolites from shrimp cultured in outdoor ponds and indoor tanks were right or left skewed. OxyHc, blood proteins, and glucose levels were higher in shrimp cultured in outdoor ponds than those observed in shrimp maintained in indoor tanks. These differences were attributed to availability of live food in outdoor culture ponds. The OxyHc/protein ratio showed that 60% of blood protein is OxyHc in outdoor-cultured shrimp and was lower than in indoor-cultured shrimp, and this ratio was lower than that found (97%) in indoor-cultured shrimp. The type of food was identified as a dominant factor affecting blood metabolites. AG in outdoor-cultured shrimp was significantly lower than that observed in indoor-maintained shrimp. Blood lactate concentration of shrimp was not affected by the culture conditions. The blood levels for protein, glucose, and lactate reported here are similar to those reported previously for other crustacean species, indicating that these can be used as a reference for evaluating the physiological status of L. vannamei. When we compared the OxyHc, blood protein, glucose, and lactate levels of L. vannamei juveniles with other crustacean species, we observed that crabs, lobsters (low-activity species), and closed-thelycum shrimp species (mean-activity species) had lower values in comparison to those observed in open-thelycum shrimp species, including L. vannamei juveniles. Possibly the blood metabolites and OxyHc could be reflecting the morphological and physiological adaptation of crustaceans, because these metabolites will depend on energetic demands.     

Geographical expansion of the invader <Emphasis Type="Italic">Caprella scaura</Emphasis> (Crustacea: Amphipoda: Caprellidae) to the East Atlantic coast

Caprella scaura (Templeton in Trans Entomol Soc Lond 1:185–198, 1836) is a native species to the western Indian Ocean. It was first described from Mauritius and later reported from several regions of the world. During the last decade, the species spread out of the Adriatic Sea, and in 2005, C. scaura was reported in Gerona, on the north-eastern coast of Spain. The present study shows the recent rapid expansion of the species to the East Atlantic. During a sampling survey of harbours along the Strait of Gibraltar (Dec 2009–June 2010), we found high densities of C. scaura associated with the bryozoans Zoobotryon verticillatum Della Chiaje, 1822, in Chipiona and Bugula neritina (L.) in Cádiz, Atlantic coast of southern Spain; on the other hand, during May 2009, an important population of C. scaura was also found in the Canary Islands (Tenerife) associated with aquaculture resources. A total of 1,034 individuals of C. scaura were studied, and we confirm that the populations are effectively established, with high densities and reproducing females during the whole year. The population from Cádiz was characterised by smaller specimens but a higher number of eggs per female. These differences could be related to environmental conditions, mainly sea water temperature and to other factors such as competition with other species, or availability of food or substrate. Significant correlation was found between female size and number of eggs in the three populations, but egg size did not vary with female body length. The morphometric relation length/width of the second gnathopod was also calculated and compared between sexes: larger males presented longer gnathopods at the three sites. The most probable introduction vector of this species is shipping; in fact, the Strait of Gibraltar is characterised by an intense shipping traffic. Also, we have observed that the native Caprella equilibra Say, 1818, is being displaced by C. scaura in the harbour of Cádiz, where this species reach densities of 35,945 ind/1,000 g of B. neritina. Further experimental studies are necessary to explore the potential danger of this species, which might spread into natural habitats too.     

Seasonal changes in biomass,growth rates and production of subantarctic calanoid copepods in the Bay of Morbihan,Kerguelen Islands

The present study addresses the ecology of two dominant copepod species in the Bay of Morbihan, Kerguelen Archipelago. The biomass of the herbivore Drepanopus pectinatus (from 2 mg m^?3 in winter up to 500 mg m^?3 in summer) is tightly coupled to seasonal changes in chlorophyll a concentration in the region, whereas the biomass of the predatory euchaetiid Paraeuchaeta antarctica increases during two distinct periods over the year: 250 mg m^?3 in early summer, with the recruitment of the annual generation, and 100 mg m^?3 in autumn, with the deposition of lipids as energy reserves in C5 stages and adults. The juvenile growth rates predicted by temperature-dependent models (0.09 day^?1) closely approximate those observed in D. pectinatusin summer, but are much greater than those observed in P. antarctica (from 0.001 to 0.04 day^?1 depending on developmental stages). This difference can be explained by the reproductive strategies and trophic positions of the two species and may also result from the dependence of larval growth on energy reserves in P. antarctica. The production rates are five- and tenfold greater in juvenile stages than in adults, respectively, for D. pectinatus and P. antarctica. The secondary production by D. pectinatusis insufficient to support P. antarcticaduring winter, when the predatory species probably shifts to alternate prey. In summer the predation by P. antarctica accounts for only a minor part of the mortality estimated for D. pectinatus (from 20% to 60% depending on the examined station). At two of the three stations examined in the Bay of Morbihan, the production of P. antarctica could potentially support the dietary requirements of planktivorous seabirds in the region (~2,000 kg prey day^?1 for common diving petrels, Pelecanoides urinatrix, and ~90 kg prey day^?1 for rockhopper penguins, Eudyptes chysocome filholi).     

Intracellular bacteria associated with the ascidian<Emphasis Type="Italic"> Ecteinascidia turbinata</Emphasis>: phylogenetic and in situ hybridisation analysis

The ascidian Ecteinascidia turbinata (Herdman) is a colonial sea squirt found in the Caribbean and Mediterranean Seas. In the present study, the bacterial complement of E. turbinata has been assessed by 16S rRNA gene analysis and the most commonly occurring strains identified by restriction fragment length polymorphism. Three strains were found to predominate using this approach, with one representing >50% of clones from both larval and adult material. The 16S rRNA gene sequence of the most commonly occurring strain did not match with any known bacterial sequences and could only be assigned to the γ-proteobacteria subdivision. The two other frequently occurring strains were assigned to the Mollicutes. In situ hybridisation analysis with eubacterial probes to 16S rRNA revealed the presence of apparently endosymbiotic bacteria in adult and larval tissue, and electron microscopy confirmed the presence of putative bacteriocytes in the larval tissue. The presence of the same bacteria in the brooded larvae suggested that they were vertically transmitted from parent to offspring. Further hybridisation using a novel probe designed to be specific to the 16S rRNA sequence of the dominant strain, highlighted the same cell types as that revealed by the eubacterial probe. The results suggest that the bacteria represent a novel strain, denoted "Candidatus Endoecteinascidia frumentensis", and that they may have an important role in the biology of E. turbinata.     

Evaluation of microelement contents in <Emphasis Type="Italic">Clethra barbinervis</Emphasis> as food for human and animals in contrasting geological areas

The young leaves of Clethra barbinervis Sieb. et Zucc, which is a deciduous tree species found in secondary forests widely in Japan, are used in spring as a local traditional food by local populations, and the bark of this plant is also preferred by sika deer, Cervus nippon. However, C. barbinervis has been known to accumulate heavy metals in its leaves. Then, we aimed to clarify the characteristics of microelement contents in C. barbinervis and to discuss the value of this species as food for humans and animals through the analysis of seasonal changes and distribution in various organs of C. barbinervis growing under two different geological conditions. We found that C. barbinervis is an accumulating and tolerant plant for Ni, Co and Mn. It accumulates Ni from serpentine soil containing Ni at high concentration, and Co and Mn from acidic soils based on crystalline schist. The seasonal variation in element concentrations in leaves indicates that the young leaves contain Cu at high concentration and that eating them in spring season may be advantageous to humans, due to the associated increase in Cu intake. The high concentrations of Cu and Zn in the bark of C. barbinervis might explain why deer prefer to eat the bark of this species.     

Effect of diet and paralarval development on digestive enzyme activity in the cephalopod <Emphasis Type="Italic">Robsonella</Emphasis><Emphasis Type="Italic">fontaniana</Emphasis>

This study aims at establishing the maturation of the digestive system in Robsonella fontaniana by means of measuring the digestive enzyme activity in paralarvae after hatching. Different groups of newly hatched paralarvae were fed with Artemia sp. (FA), Lithodes santolla zoea (FL) or were starved to serve as negative control (ST), the experimental period lasted 27 days after hatching (DAH). The semi-quantitative assay api^® ZYM (bioMérieux, France) was used to identify the pool of digestive enzymes present in paralarvae prior to selecting the specific enzymes acid phosphatase, total protease, trypsin and chymotrypsin for analysis. Mortality and larval protein content were also measured during the experiment. The survival of R. fontaniana hatchlings differed depending on the quality of the diet offered (ST < FA < FL), the same trend was observed with respect to paralarval protein content. Total protease and acid phosphatase activities were not related to either time of development or diet (P < 0.05). However, the activity of trypsin and chymotrypsin after DAH 10 was dependent on diet (P < 0.05). Paralarvae fed with Lithodes zoea (FL) performed better than paralarvae in the other two groups, exhibiting lower mortality, higher protein content and the highest proteolytic activity for trypsin and chymotrypsin. The results indicate that Lithodes-larvae are a better diet for R. fontaniana paralarvae than Artemia sp.     

Phytotoxic effects of phenolic acids from <Emphasis Type="Italic">Merostachys riedeliana,</Emphasis> a native and overabundant Brazilian bamboo

Merostachys riedeliana Rupr. is a native and overabundant bamboo species in the Brazilian Atlantic Forest. Moderate to strong allelopathic activity may be one mechanism that explains this super-dominance and the changes in structure and composition of forest areas occupied by bamboo. This study evaluated the phytotoxic effect of M. riedeliana extracts and fractions and identified their putative allelochemicals. We investigated the presence of allelochemicals in soil collected from stands occupied by M. riedeliana. Furthermore, we evaluated the putative effect of tree allelochemicals, individually and combined, on germination and growth. The aqueous extract of leaves and its ethyl acetate fraction presented the highest inhibitory effects on seed germination and seedling growth. The effect of the extracts and fractions on the target species was species-specific. Neither the individual nor the combined phenolic acids significantly inhibited seed germination; however, a pronounced growth inhibition was observed in M. bimucronata seedlings treated with vanillic acid and in E. verna and M. bimucronata seedlings treated with combined phenolic acids. Isovitexin, vitexin, isoorientin, orientin, and their O-glycoside derivatives, the lactonic dimer of the p-hydroxybenzoic acid and 3,4-methylenedioxymandelic acid were identified in the aqueous extracts and ethyl acetate fraction by Liquid Chromatography-Diode Array Dectector/Electrospray Ionization/Mass Spectrometry (LC-DAD/ESI–MS/MS). The Gas Chromatography-Mass Spectrometer (GC–MS) profile of the same extract and fraction showed the presence of benzoic, benzeneacetic, salicylic, p-hydroxybenzoic, p-hydroxyphenylacetic, vanillic, p-coumaric, protocatechuic, syringic, gallic, m-coumaric vanillylmandelic, 4-methylmandelic, 3,4-methylenedioxymandelic and trans-ferulic acids. The p-benzoic acid and the apigenin 6-C-glucoside (isovitexin) were identified in the soil extract collected from under bamboo-growing areas. Even though laboratory bioassays are not completely predictive of the allelopathic effects that occur in nature, the results of this study provide preliminary evidence of allelopathy as a possible species-specific inhibition mechanism of native species that explain the impoverishment of floristic richness and the functional groups in areas where M. riedeliana is overabundant.     

Effects of season and maternal food on<Emphasis Type="Italic"> Calanus finmarchicus</Emphasis> reproduction,with emphasis on free amino acids

Effects of maternal food environment and season were examined during spring and autumn on females, eggs and nauplii of Calanus finmarchicus, in different natural prey suspensions or cultures of Rhodomonas baltica. Females sampled in spring were in general larger, had higher protein content, and showed higher egg production and hatching rates, than in autumn. The cumulative egg production was almost double in spring compared to autumn (females fed R. baltica). Females had higher content of free amino acids (FAA) and free essential amino acids (EAA) in autumn than in the spring. Also, the FAA contents in eggs and nauplii were higher in autumn than in spring. In contrast, the composition of EAA in eggs was constant between seasons, indicating maternal regulation. The highest cumulative egg production was correlated with a high similarity in the free pool of EAA in the food suspension and the female copepod. Thus, the data support the hypothesis that similarity in the free pool of EAA of diet and female promotes high fecundity and egg hatching success in C. finmarchicus.     

A systematic study of some Black Corals species (Antipatharia,Hexacorallia) based on rDNA internal transcribed spacers sequences

A phylogenetic analysis based on rDNA internal transcribed spacers (ITS) sequences was performed on 15 species of black corals assigned to the genera Antipathes, Stichopathes, Cirrhipathes, Rhipidipathes, Antipathella, Myriopathes and Cupressopathes recorded from the Messina Strait (Mediterranean Sea) and the Bunaken Marine Park (Celebes Sea, Indonesia). The phylogenetic analysis shows that the examined species are grouped in two main branches corresponding to the families Antipathidae-Aphanipathidae and Myriopathidae. While among the Myriopathidae species there is a very small genetic distance, the Antipathidae-Aphanipathidae clade shows a high degree of divergence between different genera. According to this study, the taxon Antipathes? sp. 3 characterised by a bush-like corallum without a well-defined axis, probably belongs to a new undescribed genus of the family Antipathidae.     

Zum protonephridialsystem mariner gastrotrichen der ordnung macrodasyoidea

The protonephridia of marine gastrotrichs are described from Paradasys subterraneus Remane, Turbanella cornuta Remane, and three other species of the Turbanellidae. Both genera have several terminal organs in serial distribution. Paradasys has 6, Turbanella 4 groups of protonephridia. In Paradasys, 3 protonephridial tubules form a continuous system, while in Turbanella such a system consists of 2 to 4 (5) separate protonephridial tubules.