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.     

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.     

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.     

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.     

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.     

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.     

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.     

Temporal analysis of population genetic composition in the overexploited Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

The Japanese eel has dramatically declined in Asia since the 1970s. Over the past two decades, glass eel productivity in Taiwan has decreased, but is highly variable among each year catch, though the cause for this is unknown. The impact of both population decline and instability on genetic diversity, however, is unknown. In this study, we investigated changes in allele frequencies of Japanese eel recruitment events over the past 20 years using six polymorphic microsatellite DNA loci. Specimens of glass eels were collected yearly from a single location in northern Taiwan from 1986 to 2007. Overall genetic differentiation among all samples was very low but significant (F_ST = 0.002, P = 0.002), and only 2 out of 120 pairwise tests were significant. The relationship between genetic and temporal distance showed a slight but insignificant correlation (R² = 0.03, P = 0.0504). There were no overall significant differences in allelic richness (P = 0.35) or genetic heterozygosity (P = 0.73) among annual recruitment events. No apparent loss of genetic diversity and occurrence of a genetic bottleneck for eel populations were observed. Estimates of the effective population size (N_e) generally exceeded 500, although confidence intervals were very wide. While El Niño /Southern Oscillation (ENSO) events had little impact on genetic diversity, they may account for the annual fluctuation in glass eel catch. These results indicate long-term stability of genetic diversity in the Japanese eel with little evidence for sweepstakes recruitment.     

Fungicidal activity of Cu nanoparticles against <Emphasis Type="Italic">Fusarium</Emphasis> causing crop diseases

In India more than 60 % of the population relies on crops for their livelihood. However, crop diseases are one of the major factors limiting productivity. Hence, nanotechnology appears as a new means to control diseases and enhance yield. Here, stable copper nanoparticles were synthesized using cetyl trimethyl ammonium bromide and copper nitrate at room temperature, then characterized by UV–Visible spectrophotometry, X-ray diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy and zeta potential measurement. The antifungal activity was evaluated against three common crop pathogenic Fusarium spp. We found that stable copper nanoparticles synthesized using 0.030 M cetyl trimethyl ammonium bromide and 0.003 M copper nitrate have the maximum activity against Fusarium equiseti with a 25 mm zone of inhibition, followed by F. oxysporum (20 mm) and F. culmorum (19 mm).     

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.     

In vitro mass multiplication of screw pines (<Emphasis Type="Italic">Pandanus</Emphasis> spp.) - an important costal bio- resource

An efficient in vitro mass multiplication protocol was developed for selected species of screw pine (Pandanus fascicularis Lam., P. furcatus Roxb.). The protocol could be successfully used for large scale production of planting materials leading to cultivation and making bio-fences or green belts in coastal areas, wetlands for protection and preventing soil erosion. This will facilitate the replenishment of the bio resource and also provide good breeding space for various fauna associated with it. For developing the tissue culture protocol, shoot tips and tillers from the mature plants were used as the explants. Surface sterilization with 0.1 % mercuric chloride for 1 min. Yielded good fraction (65 %) of contamination free explants. Explants inoculated in MS solid medium supplemented with 6-benzylaminopurine (BAP, 2.5 mg/l) and indole-3-acetic acid (IAA, 0.5 mg/l), induced 2–3 shoot buds in 5 weeks. These buds either individually or along with the explant portion, sub-cultured to fresh media containing the same hormonal combinations, resulted induction of 5–6 shoot buds in 4 weeks. Repeated subcultures of shoot buds in the same medium produced 8–10 shoot buds in every 4 weeks. Shoot elongation was achieved (~ 3 cm) by transferring the shoot clusters or individual shoots to basal medium, and the elongated shoots were rooted in vitro or ex vitro. Root induction in shoots (87.4 %) was achieved in MS medium supplemented with 0.1 mg/l indole-3-butyric acid (IBA). Rooted shoots were established in paper/polythene cups filled with fine sand, which subsequently recorded 85 % establishment under the shade-net house with proper irrigation. Plants grown in poly-bags were later successfully established in different field conditions and recorded 100 % survival.     

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.     

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.     

Reproduction of<Emphasis Type="Italic"> Ophiactis resiliens</Emphasis> (Echinodermata: Ophiuroidea) in New South Wales with observations on recruitment

Reproduction of the ophiuroid Ophiactis resiliens Lyman 1879, a common species in the shallow waters around Australia, was examined over 1 year in two populations near Sydney, New South Wales. Gametogenesis was documented by histological examination of the gonads. Vitellogenesis in females and spermatocyte proliferation in males started in February and March. O. resiliens had an annual spawning period from May to September/October with peak gamete release from May to July. Thereafter, spawning occurred in some specimens through November. Gravid gonads were present for approximately 2 months longer in O. resiliens from Clovelly Bay than in those from Little Bay. At both sites spawning was episodic and gametogenesis continued through most of the spawning season. Most specimens (50–100%) collected from November to March had indeterminate/recovery stage gonads, the sex of which, could not be determined. The timing of gametogenic renewal during these months varied among individuals. O. resiliens has a 3-month planktonic stage and the presence of juveniles, 0.5 –1.5 mm disk diameter, in the coralline algal turf at Little Bay from February to May, suggests that settlement occurs in summer and autumn at this site. In contrast, juveniles were rarely encountered in coralline turf at Clovelly.     

Growth and production of<Emphasis Type="Italic"> Ophiocten gracilis</Emphasis> (Ophiuroidea: Echinodermata) on the Scottish continental slope

Growth and production of the bathyal ophiurid brittle star Ophiocten gracilis was studied from skeletal growth bands and disc size frequencies of specimens collected in sled and trawl samples taken on the continental slope off Scotland. Growth bands showed up in SEM examination as ring-like zones in surface relief and texture of the stereom microstructure of the intervertebral muscle insertions on the arm ossicles. Seasonal variability in somatic growth, presumed to underlie this growth pattern, may reflect reproduction and/or a possible non-feeding period during gonad maturation. Disc size-at-age was back-calculated from size-at-age interpreted from growth-band series on the vertebral ossicles from arms of O. gracilis. Pooled growth-band frequency data and normal-distribution mixtures based on size-at-age data were used to test for overgrowth of early growth bands on the ossicles from larger individuals. Von Bertalanffy and Gompertz growth models were fitted to the finalised back-calculated disc size-at-age data. These were used along with the modal structure of the observed disc size frequencies to develop a demographic model based on normal-distribution mixtures constrained by the growth model. These and other defining parameters were fitted by non-linear regression to size structure observed in a sample of the breeding population from 997 m depth on the Hebrides Terrace. Recruitment was estimated according to available data from sediment-trap time series. A ratio of somatic production/biomass, P_S/B, in the range of 0.43–0.54 was estimated using a fitted size/mass relationship and the increment summation method (ISM) applied to the fitted growth models. A narrower, but otherwise similar, range in estimated P_S/B ratios (0.48–0.49) was obtained in a parallel approach using the mass-specific growth rate method (MSGRM), whereby the same size/mass relationship was applied to the observed frequencies and growth parameters fitted to growth banding. Using previously obtained data on population density, a standing crop of 4.8 g wet weight (~0.58 mg AFDW) m^?2 would provide annual wet weight production in the range of 1.9–2.4 g (~0.23–0.29 mg AFDW) m^?2 in the population between ca. 700–1000 m depth. Somewhat greater production estimates (P_S/B=0.73–0.98) were obtained from MSGRM by pooling the sample with size frequencies from other large samples in which postlarval sizes were more numerous, but larger sizes less numerous. Similarly high production was estimated by MSGRM from a box corer sample from the Wyville-Thomson Ridge. Explanations for variability in size structure are discussed, but even the lower estimates are comparable to boreal shallow-water brittle stars. The high rate of growth and production by accepted deep-sea standards may be related to a capability for interface feeding.     

Attraction of the potential biocontrol agent <Emphasis Type="Italic">Galerucella placida</Emphasis> (Coleoptera: Chrysomelidae) to the volatiles of <Emphasis Type="Italic">Polygonum orientale</Emphasis> (Polygonaceae) weed leaves

Galerucella placida Baly (Coleoptera: Chrysomelidae) is a potential biocontrol agent of the rice-field weed Polygonum orientale L. (Polygonaceae). The volatile organic compound (VOC) profiles from undamaged and mechanically damaged plants, and from plants 12- and 36-h following continuous feeding of female G. placida adults and 2nd instar larvae were identified and quantified by GC–MS and GC-FID analyses. Twenty-four and 21 compounds were identified in volatiles of undamaged and insect feeding plants, respectively; whereas 22 compounds were detected in volatiles of mechanically damaged plants. Decanal and 1-dodecanol were unique to undamaged plants, and linalool was detected in volatiles of undamaged and mechanically damaged plants, but not in volatiles of insect damaged plants. However, the beetles are not attracted by none of these volatile components, when tested individually in Y-shaped glass tube olfactometer bioassays. In all plants, methyl jasmonate was predominant. 1-Undecanol was the least amount in undamaged plants, and plants 12-h after feeding by G. placida adults and larvae; whereas 1-tridecanol was the least abundant in plants 36-h after feeding by G. placida adults and larvae, and mechanically damaged plants. The beetles showed significant preference to the whole volatile blends from plants 12-h after feeding by larvae and plants 36-h after feeding by either larvae or adults compared to those of undamaged plants. Further, G. placida responded to individual synthetic compounds, 3-hexanol, 1-octen-3-ol, nonanal, and geraniol at 7, 1.38, 3.75 and 4.5 µg/25 µL CH₂Cl₂, respectively, and provide a basis for attraction of the potential biocontrol agent in the field.     

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.     

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 facile eco-friendly approach for the one-pot synthesis of 3,4-dihydro-2<Emphasis Type="Italic">H</Emphasis>-naphtho[2,3-<Emphasis Type="Italic">e</Emphasis>][1,3]oxazine-5,10-diones using glycerol as a green media

1,3-Oxazines have a wide variety of biological activities. Naphthoquinone scaffolds also exhibit several biological responses such as antithrombotic, apoptosis and lipoxygenase inhibitors. There is, therefore, a need to develop efficient green methodologies for hybridizing the two scaffolds in a single entity. Herein, we report a novel protocol for the synthesis of 3-aryl-3,4-dihydro-2H-naphtho[2,3-e][1,3]oxazine-5,10-diones by one-pot three-component condensation of 2-hydroxy-1,4-naphthoquinone, aromatic amines and formaldehyde in glycerol at 50 °C. After separation of products, the glycerol–water layer was extracted using ethyl acetate and the dried glycerol layer was successfully reused several times. The products were obtained in 85–95 % yields in 5–10 min. This environmentally benign protocol holds advantages of high yields, operational simplicity and easy workup over our earlier report.     

Movements and site fidelity of grass shrimp (<Emphasis Type="Italic">Palaemonetes pugio</Emphasis> and <Emphasis Type="Italic">P. vulgaris</Emphasis>) in salt marsh intertidal creeks

Grass shrimp are abundant residents of shallow-water habitats in US Atlantic and Gulf coast estuaries. Mark-recapture events determined the extent and direction of their movements in meso-tidal salt marsh creeks at North Inlet, South Carolina. Microwire-tagged Palaemonetes vulgaris demonstrated high fidelity for a dock in a subtidal channel. In the 48 days following release, 11 % of the 422 tagged shrimp were recaptured and 99 % of all recaptures were made there an average of 14 days post-release. Palaemonetes pugio, which favored the use of shallow intertidal areas, exhibited moderate fidelity for pools within creek beds with 53 % of the recaptured shrimp collected at the release site. Higher fidelity was observed for confluences of intertidal creeks and the subtidal channels. Of the 5565 shrimp released with coded microwire tags, 7.5 % were recaptured and 98 % of those recaptures were at release sites up to 53 days later. During another event, P. pugio released 235 m from their origin demonstrated relatively low fidelity for the new site and a lack of homing behavior. Of the 544 tagged shrimp, 3 % were recaptured; only 40 % of those recaptured were from the release site. Site fidelity is a mechanism by which previously discovered large and consistent differences in grass shrimp use of neighboring intertidal creek basins can be sustained over weeks, seasons, and years. Spatial variations and the tendency of these keystone organisms to resist relocation by tidal currents and occupy certain areas for extended periods should be considered in decisions about proposed alterations to creeks and shorelines.