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.     

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.     

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.     

Developmental and genetic evidence for the existence of three morphologically cryptic species of<Emphasis Type="Italic"> Crepidula</Emphasis> in northern Chile

The taxonomy of Crepidula has been based principally on shell morphology, but shell characteristics in this genus may be strongly influenced by the substrates they inhabit and are thus of limited use in classification. Four species in this genus are currently recognized from the Chilean coast, of which two, Crepidula dilatata and C. fecunda, are sympatric. These species are morphologically cryptic and differ only in their larval development: C. dilatata has indirect development with a planktonic phase, and C. fecunda direct development within egg capsules. The other two species, C. coquimbensis and C. philippiana, are species that occur within gastropod shells inhabited by hermit crabs and differ from the other species in internal morphology of the shell, developmental features, and geographic distribution. Analyses of nine morphological characters, eight developmental characters, and 26 enzymatic loci were carried out on four Crepidula populations in the Coquimbo region of northern Chile. Four populations of Crepidula were identified based on morphological, developmental, and protein electrophoresis features. Our results suggest the presence of C. coquimbensis plus three morphologically cryptic species including C. dilatata, C. fecunda, and a new previously unrecognized cryptic species. All four populations showed differences in larval development pattern. C. fecunda have indirect development, with free-swimming larvae. The remaining three species have intracapsular metamorphosis, but different modes of larval nutrition. C. dilatata have nurse eggs that are consumed by mechanical destruction; C. coquimbensis engulf nutritional embryos; and the new species eat nutritive embryos by rotation. Protein electrophoresis data support the specific condition of each group with different larval development and endorse the use of developmental differences in taxonomy of Crepidula. Genetic distance shows that C. dilatata and C. fecunda are closely related, and that C. coquimbensis is closer to them than to the new cryptic species.     

Purification and sequencing of a mate-recognition protein from the copepod<Emphasis Type="Italic"> Tigriopus japonicus</Emphasis>

Male copepods in the genus Tigriopus use surface proteins of female copepods to recognize appropriate mates. In a previous study of pre-copulatory mate-guarding in T. japonicus Mori, a monoclonal antibody was developed against one protein that contributed to the attractiveness of female T. japonicus as mate-guarding partners. In the present study, which was performed between December 1998 and November 2000, the antibody was used to purify two proteins from homogenates of pooled T. japonicus. A 70 kD protein was partially sequenced using mass spectrometry, revealing sequence similarity to α₂-macroglobulin. It was shown that the 70 kD protein was highly expressed in stage CV females, the most attractive stage for mate-guarding. A purified 36 kD protein was also shown to be a cognate of α₂-macroglobulin. Because the structure of both the 70 and 36 kD proteins suggested that protease activity and its inhibition might play a role in male mate choice, three protease inhibitors were tested in bioassays of male discrimination of conspecifics. The male's ability to discriminate among females was not completely blocked by the protease inhibitor, but their behavior was affected in more subtle ways. This suggests that the 70 kD protein, designated CSP70, is a variant of α₂-macroglobulin, but with more specific activity.     

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.     

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.     

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.     

Effects of phosphorus sources on volatile organic compound emissions from <Emphasis Type="Italic">Microcystis flos</Emphasis>-<Emphasis Type="Italic">aquae</Emphasis> and their toxic effects on <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>

There is diverse phosphorus (P) in eutrophicated waters, but it is considered as a crucial nutrient for cyanobacteria growth due to its easy precipitation as insoluble salts. To uncover the effects of complex P nutrients on the emission of volatile organic compounds (VOCs) from cyanobacteria and their toxic effects on other algae, the VOCs from Microcystis flos-aquae supplied with different types and amount of P nutrients were analyzed, and the effects of VOCs and their two main compounds on Chlamydomonas reinhardtii growth were investigated. When M. flos-aquae cells were supplied with K₂HPO₄, sodium pyrophosphate and sodium hexametaphosphate as the sole P source, 27, 23 and 29 compounds were found, respectively, including furans, sulfocompounds, terpenoids, benzenes, aldehydes, hydrocarbons and esters. With K₂HPO₄ as the sole P source, the VOC emission increased with reducing P amount, and the maximum emission was found under Non-P condition. In the treatments of M. flos-aquae VOCs under Non-P condition and two main terpenoids (eucalyptol and limonene) in the VOCs, remarkable decreases were found in C. reinhardtii cell growth, photosynthetic pigment content and photosynthetic abilities. Therefore, we deduce that multiple P nutrients in eutrophicated waters induce different VOC emissions from cyanobacteria, and P amount reduction caused by natural precipitation and algal massive growth results in more VOC emissions. These VOCs play toxic roles in cyanobacteria becoming dominant species, and eucalyptol and limonene are two toxic agents.     

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.     

Comparative life-history study on sympatric hyperiid amphipods (<Emphasis Type="Italic">Themisto pacifica</Emphasis> and <Emphasis Type="Italic">T</Emphasis>. <Emphasis Type="Italic">japonica</Emphasis>) in the Oyashio region,western North Pacific

Life-history features of the sympatric amphipods Themisto pacifica and T. japonica in the western North Pacific were analyzed based on seasonal field samples collected from July 1996 through July 1998, and data from laboratory rearing experiments. T. pacfica occurred throughout the year, with populations peaking from spring to summer. In contrast, T. japonica were rare from autumn to early winter, but became abundant in late winter to spring. Mature T. pacifica females and juveniles occurred together throughout the year, indicating year-round reproduction. Mature T. japonica females were observed only in spring, and juveniles occurred irregularly in small numbers, suggesting limited, early-spring reproduction in this study area. Size composition analysis of T. pacifica identified a total of eight cohorts over the 2 years of the study. Due to the smaller sample size and rarity of mature females (>9.6 mm) and males (>7.1 mm), cohort analyses of T. japonica were not comparable. Laboratory rearing of specimens at 2°C, 5°C, 8°C and 12°C revealed that a linear equation best expressed body length growth by T. pacifica, while a logistic equation best expressed body length growth by T. japoncia. Combining these laboratory-derived growth patterns with maturity sizes of wild specimens, the minimum and maximum generation times of females at a temperature range of 2–12°C were computed as 32 days (12°C) and 224 days (2°C), respectively, for T. pacifica, and 66 days (12°C) and 358 days (2°C), respectively, for T. japonica. The numbers of eggs or juveniles in females marsupia increased with female body length and ranged from 23 to 64 for T. pacifica and from 152 to 601 for T. japonica. Taking into account the number of mature female instars, lifetime fecundities were estimated as 342 eggs for T. pacifica and 1195 eggs for T. japonica. Possible mechanisms for the coexistence of these two amphipods in the Oyashio region are also discussed.Communicated by O. Kinne, Oldendorf/Luhe     

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.     

Density-dependent effects control the reproductive strategy and population growth of <Emphasis Type="Italic">Aurelia</Emphasis><Emphasis Type="Italic">aurita</Emphasis> s.l. scyphistomae

Aureliaaurita s.l. scyphistomae are capable of developing different asexual modes for propagation and thus present a multi-mode reproductive strategy. The reproduction rates and the reproductive strategy they adopt depend on a combination of various environmental parameters. We investigated the A.aurita s.l. polyp-to-polyp reproduction strategy and population growth in relation to polyp density. Our results confirmed that density-dependent factors control population growth of A. aurita s.l. scyphistomae in three different ways: (1) decreasing the polyp reproduction rate, (2) triggering the production of motile bud-like tissue particles and (3) inducing the detachment of developed scyphistomae. Whereas the decrease in the reproduction rate reduces the number of recruits, the motile particles and the detachment of scyphistomae contribute to minimizing density-dependent effects by allowing reproductive products and scyphistomae to drift away. Thus, not only are the negative effects of intraspecific competition for space and food diminished but also the potential colonization of new substrates, and further increase in scyphistoma density is favoured on larger spatial scales. The potential capability to switch its polyp-to-polyp reproduction strategy in response to environmental clues and population density may give Aurelia high adaptability in the temperate coastal waters where they commonly live and where they may be exposed to wide-ranging and fluctuating environmental variables that affect their survival and longevity. Considering these features, as well as the wide tolerance of Aurelia scyphistomae (and medusae) to environmental parameters, it is not surprising that the species/lineages of Aurelia are cosmopolitan and exhibit the most frequent bloom events worldwide.     

Intraspecific variation of photosynthesis,respiration and photoprotective carotenoids in<Emphasis Type="Italic"> Gracilaria birdiae</Emphasis> (Gracilariales: Rhodophyta)

In most rhodophytes, the quenchers zeaxanthin or lutein seem to be used for carotenoid photoprotection next to β-carotene. However, our study shows that the tropical intertidal alga Gracilaria birdiae possesses high concentrations of antheraxanthin, as well as lower concentrations of both zeaxanthin and violaxanthin. In higher plants and some algal groups, these three oxygenated carotenoids are known to protect photosystem II in a xanthophyll cycle. We were able to manipulate the carotenoid concentrations in G. birdiae by varying irradiance and temperature in laboratory experiments. Higher concentrations of violaxanthin were found in dark incubations and higher concentrations of zeaxanthin in light incubations, which is an indication that a xanthophyll cycle is active in G. birdiae. Our results also suggest a biosynthesis pathway from lycopene to β,β-carotene and β-cryptoxanthin in the light and synthesis of new lycopene in darkness. This would imply that the production of functional carotenoids is regulated by their need in photoprotection. Despite cultivation in the laboratory under the same conditions for at least 6 years, there were differences in photosynthetic properties and carotenoid dynamics between algae obtained from a population close to the Equator and another population close to the Tropic of Capricorn. These differences most probably have their origin in genetic differentiation mediated by ambient irradiance and temperature regimes. Four phycoerythrin-deficient mutant strains of G. birdiae were studied as well. Photosynthetic properties and carotenoid dynamics were basically the same in the mutants and the wild strains, but two green strains had remarkably low dark respiratory rates. This is an indication of low production of metabolites, which may be a competitive disadvantage.     

A population comparison of the strength and persistence of innate colour preference and learning speed in the bumblebee <Emphasis Type="Italic">Bombus terrestris</Emphasis>

Studies of innate colour preference and learning ability have focused on differences at the species level, rather than variation among populations of a single species. Initial strength and persistence of colour preferences are likely to affect colour choices of naïve flower visitors. We therefore study the influence of both the strength and persistence of innate colour preference (for blue) on an operant learning task (associating food reward with yellow flowers) in two populations of the bumblebee Bombus terrestris. We found that both strength and persistence of blue preference differed significantly between populations: B. terrestris dalmatinus had a weaker and less persistent blue preference than B. terrestris audax. These differences in preference also influenced learning performance. Considering only landing behaviours, one-trial learning occurred in the majority (73%) of bees, and was achieved sooner in B. terrestris dalmatinus because of its weaker blue preference. However, compared to landing behaviours the relative frequency of approach flights to rewarding and unrewarding flower types changed more slowly with task experience in both populations. When considering both approaches and landings, the rate of learning, following the first rewarded learning trial, was faster in B. terrestris audax than B. terrestris dalmatinus. However, the net effects of population differences in blue preference and learning dynamics result in similar final levels of task performance. Our results provide new evidence of behavioural differences among isolated populations within a single species, and raise intriguing questions about the ecological significance and adaptive nature of colour preference.     

Window of opportunity of <Emphasis Type="Italic">Liparis loeselii</Emphasis> populations during vegetation succession on the Wadden Sea islands

The present study aims at assessing the life span of populations of the orchid L. loeselii during successional in dune slacks on the Wadden Sea islands. An inventory of Liparis loeselii populations was carried out on 6 islands, while the population structure was assessed in more detail on the island of Texel. The occurrence of the orchid was related to soil factors and hydrological conditions. Groundwater levels were measured in a dune area with natural dune slack formation. The groundwater composition was analyses and the freshwater lenses were modelled. Various scenarios were investigated, including sea level rise and sand nourishment. The window of opportunity for L. loeselii to colonize a new slack is relatively narrow. Under favorable management, the population can survive c. 20 years. Soil pH was positively correlated with the occurrence of L. loeselii populations, while a high organic matter content was negatively correlated. Sites with large populations of L. loeselii, were situated in young dune slacks, that functioned as flow-through lakes and has top soilsinfluenced by anoxic calcareous groundwater. Modelling showed that the freshwater lens would decrease due to sea level rise, while artificial sand nourishment could counteract this effect. The populations of the Hors area on Texel, the Netherlands, can survive for several decades due to continuous formation of new slacks. Discharge of calcareous groundwater is essential to keep the pH on a high level. The hydrological systems that supply dune slack with groundwater are relatively small and are vulnerable for changes in sea level rise.     

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.     

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.     

<Emphasis Type="Italic">Corymbia</Emphasis> phloem phenolics,tannins and terpenoids: interactions with a cerambycid borer

Plant secondary chemistry mediates the ability of herbivores to locate, accept and survive on potential host plants. We examined the relationship between attack by the cerambycid beetle Phoracantha solida and the chemistry of the secondary phloem (inner bark) of two differentially attacked plantation forestry taxa, Corymbia variegata and its hybrid with C. torelliana. We hypothesised that this differential rate of attack may have to do with differences in secondary chemistry between the taxa. We found differences in the bark chemistry of the taxa, both with respect to phenolic compounds and terpenoids. We could detect no difference between bored and non-bored C. variegata trees (the less preferred, but co-evolved host). Hybrid trees were not different in levels of total polyphenols, flavanols or terpenes according to attack status, but acetone extracts were significantly different between bored and non-bored trees. We propose that variations in the bark chemistry explain the differential attack rate between C. variegata and the hybrid hosts.     

Influence of environmental factors on biodegradation of quinalphos by <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis>

Background

The extensive and intensive uses of organophosphorus insecticide—quinalphos in agriculture, pose a health hazard to animals, humans, and environment because of its persistence in the soil and crops. However, there is no much information available on the biodegradation of quinalphos by the soil micro-organisms, which play a significant role in detoxifying pesticides in the environment; so research is initiated in biodegradation of quinalphos.

Results

A soil bacterium strain, capable of utilizing quinalphos as its sole source of carbon and energy, was isolated from soil via the enrichment method on minimal salts medium (MSM). On the basis of morphological, biochemical and 16S rRNA gene sequence analysis, the bacterium was identified as to be Bacillus thuringiensis. Bacillus thuringiensis grew on quinalphos with a generation time of 28.38 min or 0.473 h in logarithmic phase. Maximum degradation of quinalphos was observed with an inoculum of 1.0 OD, an optimum pH (6.5–7.5), and an optimum temperature of 35–37 °C. Among the additional carbon and nitrogen sources, the carbon source—sodium acetate and nitrogen source—a yeast extract marginally improved the rate of degradation of quinalphos.

Conclusions

Display of degradation of quinalphos by B. thuringiensis in liquid culture in the present study indicates the potential of the culture for decontamination of quinalphos in polluted environment sites.