Site fidelity and homing behavior in juvenile loggerhead sea turtles (<Emphasis Type="Italic">Caretta caretta</Emphasis>)

To investigate site fidelity and homing behavior in juvenile loggerheads ( Caretta caretta, L.), a mark-recapture study spanning four years (1998–2001) was conducted in Core Sound, N.C., USA. Each year of the study, approximately half of the turtles captured were tagged and released near the capture sites ( n=207), while the remaining turtles were displaced 15–20 km and released ( n=198). Loggerheads in both groups were recaptured in equal proportions near the original capture sites and many individuals were also recaptured in subsequent years. These data imply that juvenile loggerheads often returned to their capture sites following displacement, because if turtles dispersed randomly or remained near their release sites, then fewer displaced turtles should have been caught again. Moreover, because turtles migrate out of North Carolina sounds each winter, turtles recaptured at the same locations in different years evidently returned to specific sites following long migrations. To further investigate homing behavior, a small number of displaced turtles ( n=28) were tracked using radio telemetry following their release. Although transmitters detached from most turtles within a few days, analyses of initial headings showed strong orientation in the direction of the capture site. In addition, four turtles successfully tracked for longer periods of time all returned rapidly to the vicinity of the capture location and remained in the area. Taken together, the results of this study indicate that juvenile loggerheads exhibit fidelity to specific areas during summer months and possess the navigational abilities to home to these areas following forced displacements and long-distance migrations.     

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.     

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.     

The reproductive cycle of <Emphasis Type="Italic">Scolelepis goodbodyi</Emphasis> (Polychaeta,Spionidae)

The aim of this study was to examine the reproductive and gametogenic cycle of the spionid polychaete Scolelepis goodbodyi (Jones). Every 15 days, in the upper level of the intertidal zone on Barequeçaba Beach in southeastern Brazil, a 100 m² area was delimited and within it three points were selected and sampled using a core sampler 0.01 m² in area × 0.2 m long. Each sample was divided into three sub-samples: surface (2 cm) and middle (8 cm), which were sieved through 0.5, 0.25 and 0.125 mm mesh sieves; and lower (10 cm), sieved through 1.0 and 0.5 mm mesh sieves. Males, females and undetermined individuals were separated, and the width of the third setiger was measured. The gonadal development of each individual was classified based on external features of the gametogenic setigers. Ten to 20 adults from each sampling period were classified according to their gonadal development stage and then analyzed histologically for gametogenic studies, for comparison with the former classification. Some of the ripe females were used for fecundity evaluation. The diameters of the oocytes were measured prior to spawning. Eight hundred and twenty-eight females (29.70%), 848 males (30.42%) and 1,112 undetermined individuals (39.89%) were analyzed. The sex ratio did not differ significantly from unity (χ² = 0.24; df = 1; P < 0.05). Males and females were more abundant in May 2002 (ca. 80%) and between November 2002 and May 2003 (between 77 and 90%); the proportion of undetermined individuals did not represent recruitment alone, but also a high incidence of adults with recovering gonads. Fecundity varied from 134 oocytes in a 0.54 mm W3 individual to 289 oocytes in a 0.4 mm W3 individual, with a mean of 220 (SD = 57) oocytes per female and 19 (SD = 5) oocytes per gametogenic setiger. Oocytes were ellipsoid, and ranged in size from 150 × 80 (to 220 × 120 μm (mean ± SD = 173 × 113 ± 15 × 11 μm). Neither a well-defined gametogenic cycle nor synchrony in the reproductive period was observed, because many gonadal development stages occurred in each month. Nevertheless, peaks of maturity were observed between October–December 2002 and April–May 2003.     

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).     

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.     

<Emphasis Type="Italic">Bacillus algicola</Emphasis> decolourises more than 95% of some textile azo dyes

Textile effluents in natural waters pose environmental health problems if not treated to safe limits. Various bacterial species have the potential to degrade dyes. Here we studied the ability of Bacillus algicola to decolourise red, blue and yellow azo dyes. B. algicola was isolated from soil samples taken from a sanitary landfill site. Isolation and screening were performed using mineral salt medium. Dye-decolourising isolates were assessed in their capacity to decolourise dyes. Experiments were conducted at pH 6, 7 and 8, and 25, 35 and 45 °C. Phytotoxicity of the dyes and biodegradation products was assessed by seed germination tests. Results show that B. algicola gave the highest decolourisation at pH 8.0 and 25 °C in the presence of yeast extract as media supplement. B. algicola degraded the red and blue azo dyes by over 95%. The phytotoxicity results indicated that biodegradation products of the red and blue azo dyes were not toxic. Biodegradation products of the yellow dye were, however, toxic and considerably hindered germination. From these results, we infer that B. algicola has good potential for degrading and decolourising the red and blue test azo dyes.     

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.     

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.     

Ecological success of the cladoceran <Emphasis Type="Italic">Penilia avirostris</Emphasis> in the marine environment: feeding performance,gross growth efficiencies and life history

Marine cladocerans are important contributors to the zooplankton community of tropical and temperate coastal ecosystems during the warmer months, when they show explosive population growth. Despite this fact, little information is available on their ecology compared with the extensive studies on their freshwater relatives. The main objective of this study was to determine the in situ feeding and growth rates, and life history parameters of Penilia avirostris in São Sebastião Bay (Brazil) during austral summer 2004, as a premise to understand the advantages of this cladoceran in oligotrophic waters. Culture development experiments, monitored for a period of 12 days, showed that maximum juvenile release occurred after 2 days, and that the development duration of a complete cohort was around 9 days. From bottle incubation grazing experiments, significant ingestion rates upon flagellates, ciliates, dinoflagellates and diatoms were detected. Flagellates were the most important contributors to P. avirostris diet (ca. 80%). P. avirostris ingested between 28 and 97% of its own carbon biomass per day (daily ration) and individual growth rates of this marine cladoceran (0.10–0.24 d^?1) increased with prey availability. The combination of ingestion rates of natural prey and growth rates provided gross growth efficiencies (GGE) of 15–53%, on a carbon basis. Our results suggest that P. avirostris has similar GGE to copepods, although at low food conditions the values for the marine cladocerans seems slightly higher. However, this characteristic alone does not explain the explosive growth and community dominance shown by P. avirostris. Therefore, other traits related to the reproductive biology of the species, such as short generation time, parthenogenetic reproduction, and continuous somatic growth, seems to be mostly responsible for the success of P. avirostris in many marine ecosystems during their seasonal occurrence.     

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.     

New beach in a shallow estuarine lagoon: a model-based <Emphasis Type="Italic">E. coli</Emphasis> pollution risk assessment

A 3D hydrodynamic model has been applied to the Curonian Lagoon to study the pollution impact of E. coli on a new beach that might be opened in the lagoon. Through a field survey the E. coli inputs were measured and then used in the numerical model, and through laboratory experiments the decay rate of E. coli was established. The model has been calibrated and validated for the year 2015, and several scenarios have been studied, such as sewage system breakdown, severe weather conditions or high river loads. The model has then been run for a period of 12 years to obtain a robust statistics for the pollution on the planned beach. Results show that the decay rate of E. coli is between 0.55 days and 2.3 days and the modeled decay times are compatible with these numbers. The only scenario that would create a risk for the bathing waters of the beach is a breakdown of the sewage system on the Curonian Spit. In this case the hours (and days) over legally allowable bathing threshold were computed in order to estimate the number of days the beach could be closed. These results have been confirmed by the 12 year simulations. With an influence map analysis the two most critical sewage systems could be identified.     

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.     

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.     

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.     

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.     

Effect of arsenic species on the growth and arsenic accumulation in <Emphasis Type="Italic">Cucumis sativus</Emphasis>

The effects of arsenic (As) species, such as As(III), As(V) and dimethylarsinic acid (DMA), on the accumulation of As in cucumber (Cucumis sativus), as well as on its growth in a soil mesocosm were evaluated. When Cucumis sativus was cultivated in soils contaminated with 20 and 50 mg/kg of As(III), As(V) or DMA for 40 days, the growth was markedly inhibited by the inorganic As (As(III) and As(V)) rather than the organic As (DMA). Irrespective of the As species, the As concentrations accumulated in Cucumis sativus increased with increasing As concentration in the soil. The As bioaccumulation factors from soil into the tissue of Cucumis sativus were 17.5–35.4, 29.3–42.7 and 17.6–25.7 for As(III), As(V) and DMA, respectively. In addition, the As translocation factors from the roots to shoots were 0.025–0.031, 0.018–0.032 and 0.014–0.026 for As(III), As(V) and DMA, respectively. In conclusion, Cucumis sativus mainly accumulated As in its roots rather than its shoots and easily accumulated inorganic rather than organic As from the soil into its tissue.     

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.     

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.