Chemical resistance of gorgonian corals against fungal infections

The frequency and impact of diseases affecting corals throughout the Caribbean have been increasing but little is known about the factors promoting the emergence and outbreak of disease. A disease caused by a fungal pathogen [Aspergillus sydowii (Thom et Church)] which affects Caribbean sea fan corals provided an opportunity to examine the efficacy of coral crude extracts in disease resistance. Minimum inhibitory concentration (MIC) assays showed that of the 20 common gorgonian species in the Florida Keys, extracts from 15 species had MICs < 15 mg ml⁻¹ against A. sydowii pathogenic to sea fans. Extracts from several species in two gorgonian genera (Pseudoplexaura and Pseudopterogorgia) were among the most active, with MICs < 10 mg ml⁻¹. Gorgonia ventalina L., one of two sea fan species known to be hosts to A. sydowii in the field, had an MIC < 10 mg ml⁻¹, suggesting that complete disease resistance requires more active extracts. For the antifungal compounds to be effective in situ, they must also occur in sufficiently high concentrations in living coral tissue. For example, Pseudopterogorgia americana (Gmelin) had comparatively potent extracts but did not have sufficient concentrations in the tissue to be effective. Conversely, Plexaura homomalla Esper extracts were less potent but occurred in high enough concentrations in the tissue to be effective against A. sydowii. When potency and extract concentration are considered together (i.e. potency × concentration), several other gorgonian corals emerge as likely hosts to A. sydowii. Crude extracts from the most active gorgonian species were also effective against two geographic variants of A. sydowii pathogenic to sea fans, a non-pathogenic terrestrial strain of A. sydowii, and three strains of A. flavus Link known to be human, plant, and insect pathogens (MIC range, 7.5 to > 15 mg ml⁻¹). Although the potency in these assays did not attain a clinically significant level, the potency is comparable to a known antifungal agent, hygromycin B, which had an MIC ≤7.5 mg ml⁻¹ in our assays, highlighting the potential of these gorgonian corals for bioprospecting. Received: 29 May 1999 / Accepted: 22 May 2000     

Exploring the role of chitinolytic enzymes in the sea fan coral, <Emphasis Type="Italic">Gorgonia ventalina</Emphasis>

Chitinases are involved in defense against chitinaceous pathogens in both invertebrates and vertebrates. This study investigated whether sea fan corals, Gorgonia ventalina (Linnaeus) collected from the Florida Keys between the summer of 2002 and the summer of 2005 contain chitinases, and whether these enzymes could serve an analogous protective role against the fungal pathogen, Aspergillus sydowii (Thom et Church). Crude extracts of healthy sea fans contained detectible levels of exochitinase activity in an in vitro microplate assay using fluorogenic substrates. The exochitinase levels decreased upon injury, agitation, or manipulation of the tissue. A concurrent, transient increase of exochitinase in the surrounding water suggests that sea fans release chitinases as a response to these stresses. By contrast, endochitinase was detected in only 2 of 15 sea fans (13%), suggesting a high degree of variation for this enzyme. Sea fan chitinase-containing seawater and anion exchange chromatography fractions were both active against A. sydowii in an absorbance-based antifungal assay. The presence of chitinases in sea fan extracts, their release into the surrounding water upon stress, and their activity against A. sydowii suggests that further study of these enzymes in coral stress responses is warranted.     

Water-soluble extracts of the diatom Thalassiosira rotula induce aberrations in embryonic tubulin organisation of the sea urchin Paracentrotus lividus

Eggs and embryos of the sea urchin Paracentrotus lividus were used as a model to study the effect at the cellular level of potential anti-mitotic compounds extracted from the diatom Thalassiosira rotula. Eggs and embryos incubated in a water-soluble diatom extract, corresponding to 5 × 10⁶ and 10⁷ cells ml⁻¹, were totally blocked (i.e. cell division was blocked) at the one-cell stage. At lower concentrations (2.5 and 1.25 × 10⁶ cells ml⁻¹), the first mitotic division was inhibited in 32 ± 26% and 25 ± 3.5% of the zygotes, respectively, demonstrating the dose-dependent effect of diatom extracts on sea urchin development. Immunofluorescence dyes, specific for DNA and α-tubulin subunits, were used to stain nuclei and microtubules in sea urchin embryos during various phases of development. Images with the confocal laser scanning microscope showed that tubulin was not organised in filaments at the sperm aster and cortex levels, and that the pronuclei were not fused in embryos incubated soon after fertilisation with water-soluble diatom extracts corresponding to 10⁷ cells ml⁻¹. At lower diatom-extract concentrations (4 × 10⁶ cells ml ⁻¹), fusion of the pronuclei occurred but the mitotic spindle was not formed. Microtubules were clearly de-polymerised and the chromatin appeared globular and compacted at the centre of the cell. A similar structure was observed for sea urchin embryos incubated with 0.1 mM colchicine, a potent anti-mitotic compound. When sea urchin embryos were incubated in water-soluble diatom extracts at different times prior to the first mitotic division, microtubules appeared de-polymerised at each step, from pronuclear fusion to telophase, and cell division was blocked. At the histological level, embryos incubated with 4 × 10⁶ cells ml⁻¹ diatom extract showed nuclear fragmentation without cytokinesis. The possible use of sea urchin embryos as a bioassay to test for other unknown compounds with cytotoxic activity in phytoplankton species is discussed. Received: 7 May 1998 / Accepted: 9 December 1998     

Effects of inorganic and organic phosphates on feeding, feeding absorption, nutrient allocation, growth and righting responses of the sea urchin Lytechinus variegatus

The sea urchin Lytechinus variegatus is capable of surviving chronic exposure to sodium phosphate (inorganic phosphate) concentrations as high as 3.2 mg l⁻¹, and triethyl phosphate (organic phosphate) concentrations of 1,000 mg l⁻¹. However, chronic exposure to low (0.8 mg l⁻¹ inorganic and 10 mg l⁻¹ organic phosphate), medium (1.6 mg l⁻¹ inorganic and 100 mg l⁻¹ organic phosphate) or high (3.2 mg l⁻¹ inorganic and 1,000 mg l⁻¹ organic phosphate) sublethal concentrations of these phosphates inhibits feeding, fecal production, nutrient absorption and allocation, growth and righting behavior. Food consumption and fecal production declined significantly in individuals exposed to medium and high concentrations of inorganic phosphates and all levels of organic phosphates. Feeding absorption efficiencies for total organics and carbohydrates decreased significantly in individuals held in the highest concentration of organic phosphate. Feeding absorption efficiencies for lipids were significantly reduced in the highest inorganic phosphate concentration only, while they decreased significantly for protein with increasing phosphate exposure. Carbohydrate and lipid levels in gonad and gut tissues decreased significantly with exposure to increasing phosphate concentrations, potentially impairing both gametogenesis and nutrient storage in the gut. Moreover, gonad indices significantly decreased in individuals exposed to the highest concentrations of either phosphate. Growth rates decreased significantly under the influence of all phosphate concentrations, while increasing in seawater alone. Individuals exposed to increasing phosphate concentrations showed reduced righting responses (a measure of stress) and no acclimation in righting times during chronic exposure to phosphates over a 4 week period. These findings indicate that shallow-water populations of L. variegatus subjected to inorganic and organic phosphate pollutants will exhibit stress and be inhibited in their growth and performance due to reductions in feeding, nutrient absorption and allocation of nutrients to key somatic and reproductive tissues. Received: 10 April 2000 / Accepted: 2 October 2000     

Elemental composition of a tusk of a dugong, Dugong dugon, from Exmouth, Western Australia

Element analysis of the annual increments in a longitudinal section from the tusk of a female dugong Dugong dugon (Müller) from Exmouth, Western Australia, was carried out by X-ray fluorescence-imaging, inductively coupled plasma-atomic emission spectrometry (ICP-AES) and inductively coupled plasma-mass spectrometry (ICP-MS). Nine elements (Ba, Ca, Fe, Li, Mg, Na, P, Sr and Zn) were present in concentrations amenable to determination by these techniques. Most elements revealed both long-term trends and shorter, year-to-year, variations. For example, sodium concentrations increased from 0.55 to 0.72% with the age of the dugong. Strontium concentrations (mean 0.17%) were closely correlated (r = 0.86) with those of barium (mean 4.5 mg kg⁻¹). Zinc concentrations increased from ∼70 to 170 mg kg⁻¹ with dugong age, but also showed shorter-term fluctuations of ∼30 mg kg⁻¹ that were correlated (r = 0.41) with mean annual Fremantle sea level (a measure of the Southern Oscillation Index and strength of the Leeuwin Current). The concentrations of the elements and correlations with year and between pairs of elements are discussed. Received: 10 January 1997 / Accepted: 21 April 1997     

Growth and productivity of the high Antarctic Bryozoan Melicerita obliqua

We analysed growth of the Antarctic bryozoan Melicerita obliqua (Thornely, 1924) by x-ray photography and stable isotope analysis. M. obliqua colonies form one segment per year, thus attaining maximum length of about 200 mm within 50 years. In the Weddell and Lazarev Seas, annual production/biomass ratio of M. obliqua is 0.1 yr⁻¹, which is in the range of other Antarctic benthic invertebrate populations. Production amounts to 3.34 mg C_orgm⁻² yr⁻¹ and 90.6 mg ash m⁻² yr⁻¹ on the shelf (100 to 600 m water depth), and to 0.13 mg C_orgm⁻² yr⁻¹ and 36.8 mg ash m⁻² yr⁻¹ on the slope (600 to 1250 m water depth). Received: 27 February 1998 / Accepted: 8 May 1998     

Use of the fluorochrome calcein as an in situ growth marker in the brown mussel Perna perna

This study assesses the potential of the fluorochrome calcein for use as a growth marker in bivalve shell growth studies. Calcein solutions were administered in situ to the brown mussel Perna perna (Linneaus), both by injection and immersion, and the effect of calcein concentrations on fluorescent mark deposition and mussel mortality was investigated in the laboratory. Field investigations showed that, 1 month after administration, calcein injection (125 mg l^−l) into the mantle cavity produced superior results to the immersion treatments (150 and 500 mg l^−l). Both methods resulted in fluorescent mark incorporation at the growing edge, but during immersion general calcein deposition associated with endolith activity resulted in fluorescence that made identification of a distinct datum point difficult. In contrast, the injection method produced clearly defined growth marks, which were easily distinguished from autofluorescence and persisted without visible degradation for a minimum of 9 months. Shell growth rates estimated using the fluorescent mark as a datum point were similar to those from earlier studies using different methods. Laboratory investigations revealed that at␣calcein concentrations of 80 mg l^−l and above, 100% of juvenile (20 to 30 mm) and adult mussels (60 to 70 mm) retained a visible growth mark, while at concentrations >160 mg l^−l all marks were bright and clearly defined. No mussel mortality was exhibited at any time, even at calcein concentrations of 640 mg l^−l, eight times higher than those required for mark deposition. These results suggest that, compared to traditional methods of bivalve growth determination, the use of fluorochromes presents a relatively inexpensive, non-invasive and rapid alternative. When using calcein as a growth marker, problems associated with some other fluorochromes (e.g. inconsistent mark incorporation, high post-treatment mortality) were not exhibited. Received: 14 July 1998 / Accepted: 23 October 1998     

Light and carbon balance in the seagrass Thalassia testudinum : evaluation of current production models

The production dynamics and carbon balance of Thalassia testudinum in the lower Laguna Madre, Texas, USA, were examined during the 1995 summer period based on in situ photosynthesis vs irradiance (PI) measurements and continuous measurements of underwater photon-flux density (PFD). The validity of applying the H _sat model, used to calculate production for Zostera marina as the product of the maximum rate of photosynthesis (P _max) and daily hours of saturating irradiance (H _sat) was assessed for T. testudinum by comparison with integrated production estimates derived through numerical integration. Gross integrated production values were combined with dark-respiration measurements of photosynthetic (PS) and non-photosynthetic (NPS) tissues and areal biomass to generate daily whole-plant carbon balance. Production and whole-plant carbon balance are discussed in relation to surface and underwater PFD measurements, biomass and other physical and chemical parameters collected during a 1 yr period from January to December 1995. The H _sat model significantly underestimated production during all summer months, averaging 70% of integrated production over the entire study period. Gross integrated production ranged between 11.5 mg C g⁻¹ leaf dry wt d⁻¹ in June (during a period of unseasonably low PFDs caused by a drift-alga mat covering the seagrass bed) to 26.7 mg C g⁻¹ leaf dry wt d⁻¹ in July. Modeled net carbon gain was highest in July at 454 mg C m⁻² d⁻¹ (1.4 g dry wt m⁻² d⁻¹), sufficient to account for measured rates of leaf production in the study area and representative of T. testudinum populations of low productivity. During part of the summer period, however, the population was in negative carbon balance. The relatively low productivity of this population and the periods of negative carbon balance are attributed to low net photosynthesis:dark respiration (P _net:R _d) ratios, sporadic low-light periods, the small fraction of PS tissue relative to whole-plant biomass (5 to 13%) and nutrient limitation. Production models are sensitive to both light availability and the proportion of PS tissue supporting NPS biomass as reflected in whole-plant P _net:R _d ratios. Received: 13 August 1997 / Accepted: 6 March 1998     

Feeding and assimilation kinetics of Artemia franciscana fed Isochrysis galbana (clone T. Iso)

Artemia franciscana was grown on Isochrysis galbana Green (clone T. Iso) at saturated food concentrations (13 to 20 mg C l⁻¹) for 11 d at 26 to 28 °C, and 34 ppt salinity. Three groups of brine shrimp were used in the feeding experiments: metanauplius III and IV (Group 1), post-metanauplius II and III (Group 2) and post-metanauplius VIII (Group 3), corresponding to 4-, 7- and 11-d-old animals, respectively. The ingestion rate, clearance rate and carbon balance were estimated for these stages at different concentrations of ¹⁴C-labeled I. galbana ranging from 0.05 to 30 mg C l⁻¹. The handling time of algae was determined for all three groups. The ingestion rate (I, ng C ind⁻¹ h⁻¹) increased as a function of animal size and food concentration. In all three groups, the ingestion rate increased to a maximum level (I _max) and remained constant at food concentrations ≥10 mg C l⁻¹ (saturated food concentrations). The clearance rate (CR, μl ind⁻¹ h⁻¹) increased with increasing food concentration up to a maximum rate (CR _max), after which it decreased for even higher food concentrations. The functional response of A. franciscana was most consistent with Holling's Type 3 functional response curve (sigmoidal model), which for the two oldest groups (Group 2 and 3) differed significantly from a Type 2 response (p < 0.05). The gut passage time for the three groups of A. franciscana, feeding on saturated food concentration (20 mg C l⁻¹), varied between 24 and 29 min. As the nauplii developed to pre-adult stage the handling time of the algae increased as a function of animal size. The assimilation rate (ng C ind⁻¹ h⁻¹) in the youngest stages (Group 1 and 2) increased with increasing food concentrations, reaching a maximum level close to 10 mg C l⁻¹. At higher food concentrations the assimilation rate decreased, and the proportions of defecated carbon increased, reaching 60 to 68% in the post-metanauplius stages (Group 3). The assimilation efficiency (%) was high at the lowest food concentrations in all three groups (89 to 64%). At higher concentrations, the assimilation efficiency decreased, reaching 56 to 38% at the highest concentrations. Received: 2 February 2000 / Accepted: 25 March 2000     

Density,size structure and aspergillosis prevalence in <Emphasis Type="Italic">Gorgonia ventalina</Emphasis> at six localities in Puerto Rico

Gorgonia ventalina’s density, size structure and lesion prevalence was measured at six sites in Puerto Rico that exhibited variation in horizontal water transparency, sedimentation rates, suspended particle matter, scleractinian and macroalgal cover. G. ventalina density varied significantly among sites (between 0.84 and 0.007 colonies/m²), and was positively correlated with water transparency. Size structure did not vary much among sites, and reflects high mortality among the smaller size classes and high survivorship in large colonies. Prevalence of active fungi-induced lesions (type I) did not vary significantly among sites and was density-independent. However, prevalence of old lesions of unknown origin (type II) did vary among reefs and was negatively correlated with water transparency. Prevalence of types I or II lesions was independent of colony size. Our results suggest that (1) turbidity and sedimentation are important abiotic factors controlling the abundance of sea fans, (2) variation in settlement success and early survivorship of recruits has more impact on the sea fan populations than variation in the survivorship of large colonies and (3) prevalence of aspergillosis (type I) at the studied sites is similar to that reported for other Caribbean reefs and supports the epizootic nature of the disease and (4) lesions with exposed skeleton are more likely to be colonized by fouling organisms at impacted reefs. The combined effects of anthropogenic impacts and aspergillosis may cause local extinctions of sea fans, as is becoming evident in many reefs in Puerto Rico.     

Fluctuations of the red tide flagellates Chattonella spp. (Raphidophyceae) and the algicidal bacterium Cytophaga sp. in the Seto Inland Sea, Japan

A marine algicidal gliding bacterium Cytophaga sp. strain J18/M01 was isolated in 1990 from a station in northern Harima-Nada, the Seto Inland Sea, Japan, using the harmful red tide alga Chattonella antiqua (Hada) Ono as a susceptible organism. The bacterium can prey upon various species of microalgae. Temporal fluctuations of this bacterium and Chattonella spp. [C. antiqua and C. marina (Subrahmanyan) Hara et Chihara] were investigated weekly at the above station in the summer of 1997 and 1998, using immunofluorescence assay employing highly specific polyclonal antibodies for the bacterium. In the summer of 1997, the cell density of Chattonella spp. showed a maximum value (70 cells ml⁻¹) on 8 July, and decreased thereafter. The bacterium Cytophaga sp. J18/M01 was commonly detected around a few hundreds of cells per milliliter or less. The number of Cytophaga sp. J18/M01 increased after the peak of Chattonella spp., and the maximum cell number of the bacterium was 1350 ml⁻¹. This algicidal bacterium also followed the changes of total amounts of microalgal biomass (chlorophyll a+pheophytin) when Chattonella spp. were absent. In the summer of 1998, Chattonella spp. were relatively less abundant (maximum 21 cells ml⁻¹), and the algicidal bacterium Cytophaga sp. J18/M01 showed a close relationship with the change of total microalgal biomass. The present study suggests that the algicidal bacterium Cytophaga sp. J18/M01 preyed upon, not only harmful red tide microalgae, but also other common microalgae such as diatoms, and the bacterium presumably plays an important role in regulating microalgal biomass in natural marine environments. Received: 20 April 2000 / Accepted: 1 December 2000     

Spatial and temporal pattern in seagrass community composition and productivity in south Florida

We document the distribution and abundance of seagrasses, as well as the intra-annual temporal patterns in the abundance of seagrasses and the productivity of the nearshore dominant seagrass (Thalassia testudinum) in the south Florida region. At least one species of seagrass was present at 80.8% of 874 randomly chosen mapping sites, delimiting 12,800 km² of seagrass beds in the 17,000-km² survey area. Halophila decipiens had the greatest range in the study area; it was found to occur over 7,500 km². The range of T. testudinum was almost as extensive (6,400 km²), followed by Syringodium filiforme (4,400 km²), Halodule wrightii (3,000 km²) and Halophila engelmanni (50 km²). The seasonal maxima of standing crop was about 32% higher than the yearly mean. The productivity of T. testudinum was both temporally and spatially variable. Yearly mean areal productivity averaged 0.70 g m⁻²day⁻¹, with a range of 0.05–3.29 g m⁻² day⁻¹. Specific productivity ranged between 3.2 and 34.2 mg g⁻¹ day⁻¹, with a mean of 18.3 mg g⁻¹ day⁻¹. Annual peaks in specific productivity occurred in August, and minima in February. Integrating the standing crop for the study area gives an estimate of 1.4 × 1011 g T. testudinum and 3.6 × 10¹⁰ g S. filiforme, which translate to a yearly production of 9.4 × 10¹¹ g T. testudinum leaves and 2.4 × 10¹¹ g S. filiforme leaves. We assessed the efficacy of rapid visual surveys for estimating abundance of seagrasses in south Florida by comparing these results to measures of leaf biomass for T. testudinum and S. filiforme. Our rapid visual surveys proved useful for quantifying seagrass abundance, and the data presented in this paper serve as a benchmark against which future change in the system can be quantified. Received: 30 January 2000 / Accepted: 24 July 2000     

Growth and grazing responses of tintinnid ciliates feeding on the toxic dinoflagellate Heterocapsacircularisquama

Growth and feeding rates of two tintinnid species, Favellaazorica and Favellataraikaensis, were determined under various concentrations of the dinoflagellate Heterocapsacircularisquama which has been reported as highly toxic to shellfish. Mean growth rates of F. azorica and F. taraikaensis on a diet of H.circularisquama (ca. 10² cells ml⁻¹) were 2.15 and 1.97 doublings d⁻¹, respectively. These values are similar to those on a diet of Heterocapsatriquetra which is suitable food for various zooplankton. However, growth rates of both tintinnid species decrease with increasing concentrations of >10³ cells ml⁻¹ of H. circularisquama. In particular, H. circularisquama under conditions of >10³ cells ml⁻¹ caused mortality in F.taraikaensis, probably due to toxins. Clearance and ingestion rates of F. azorica on H. circularisquama were 4.1 to 27.5 μl ind⁻¹ h⁻¹ and 1.5 to 28.7 cells ind⁻¹ h⁻¹, respectively, at concentrations of <10⁴ cells ml⁻¹ and those of F. taraikaensis were 0.9 to 22.1 μl ind⁻¹ h⁻¹ and 0.1 to 13.0 cells ind⁻¹ h⁻¹, respectively, at concentrations of <10³ cells ml⁻¹. Both clearance and ingestion rates on H.circularisquama were higher for replicates fed on H.triquetra. Daily grazing impact of the two species of Favella on the initial stage of a bloom of H.circularisquama were estimated to reach 6 to 50% of H. circularisquama at a concentration of 540 cells ml⁻¹, indicating that grazing by tintinnids such as Favella spp. may significantly regulate the initial stages of blooms of H. circularisquama. Received: 3 January 1997 / Accepted: 27 January 1997     

Host range and resistance to aspergillosis in three sea fan species from the Yucatan

Knowledge of host range and mechanisms of disease resistance is fundamental to predicting impacts and spread of marine diseases. Prevalence of signs of aspergillosis, caused by the terrestrial fungus Aspergillus sydowii, was assessed in the Yucatan among three species of sea fan: Gorgonia ventalina, G. cf. mariae, and G. flabellum. The Yucatan is unusual in that ranges of all three sea fan species overlap at many sites along a cline of increasing depth, allowing us to evaluate potential causes of differing prevalence among species. Signs of aspergillosis were observed in all the three species. However, the prevalence of infection in G. cf. mariae, a deep-water species, was consistently low even at sites where G. ventalina was common and had moderate levels of infection. Because G. cf. mariae is a relatively small-stature sea fan, we compared the prevalence of signs in G. cf. mariae to a subset of comparatively sized G. ventalina. G. ventalina had a significantly higher prevalence of aspergillosis, indicating that size does not explain the lower prevalence in G. cf. mariae. Prevalence of disease signs on the shallow-water G. flabellum was also significantly higher than G. cf. mariae, but did not differ from G. ventalina. To test the hypothesis that higher chemical resistance accounts for the low prevalence of disease in G. cf. mariae, we measured the response of A. sydowii in culture to antifungal extracts from each sea fan species. Significantly lower fungal growth rates on extracts of G. cf. mariae than G. ventalina support the hypothesis that G. cf. mariae is more chemically resistant to aspergillosis. When comparing sea fan disease across different regions of the Yucatan, we detected significantly higher prevalence in G. ventalina near Akumal than further north near Cozumel and Puerto Morelos. In Akumal, there was a strong positive correlation between sea fan size and disease, with the largest fans showing the highest prevalence and severity in all three species. In addition, prevalence of aspergillosis in G. ventalina was density-dependent in Ak.     

Seasonal variations in hepatopancreas fatty acid profiles of two colour forms of shore crabs, Carcinus maenas

 The relationship between intermoult duration (coloration), sex, size and seasonal variations in fatty acid (FA) profiles was studied in a population of shore crabs, Carcinus maenas, inhabiting the Isefjord, Denmark. For male shore crabs, the total hepatopancreas FA content was high in July and December (12.7 to 16.0 mg g⁻¹ dry weight, dw) but lower in May and September (7.3 to 10.0 mg g⁻¹). This indicates that male shore crabs are in relatively good condition before winter, when the crabs migrate off shore, but in relatively poor condition when they return to shallow waters during spring. The hepatopancreas FA content also decreased over the mating season. After the mating season the hepatopancreas FA content of males had decreased to approximately 60% of that prior to the mating season. Female shore crabs had significantly higher hepatopancreas FA levels than males in May (11.7 mg g⁻¹ dw), September (12.6 mg g⁻¹ dw) and December (17.9 mg g⁻¹ dw) but lower levels in July (9.5 mg g⁻¹ dw). This indicates that the spawning season is the most energy-demanding part of the female reproductive cycle. For all seasons, the hepatopancreas FA content of green shore crabs was significantly higher than that of red shore crabs. For both colour forms, the amount of polyunsaturated fatty acids (PUFAs) was significantly higher than that of saturated fatty acids (SAFAs) and monounsaturated fatty acids (MUFAs), with the relative proportion of PUFAs increasing when the total hepatopancreas FA content decreased. For both genders and colour forms, the most dominating SAFA was palmitic acid (16:0). Palmitoleic acid (16:1ω7), vaccenic acid (18:1ω7) and oleic acid (18:1ω9) were the three MUFAs found in highest concentrations. The most dominating PUFA was eicosapentaenoic acid (EPA, 20:5ω3). Docosahexaenoic acid (DHA, 22:6ω3) and arachidonic acid (AA, 20:4ω6) were also abundant in all samples. The results demonstrated that season, sex, size and intermoult duration all influence the amount of FAs present in the hepatopancreas of shore crabs. Received: 23 September 1999 / Accepted: 29 April 2000     

Experimental investigation into the effects of suspended sediment on fertilisation, larval survival and settlement in a scleractinian coral

Laboratory and field experiments were used to determine whether high (≃100 mg l⁻¹), low (≃50 mg l⁻¹) and control (≃0 mg l⁻¹) levels of suspended sediment affected fertilisation, larval survival, and larval settlement in the scleractinian coral Acropora digitifera (Dana, 1846). Both high- and low-sediment treatments significantly decreased fertilisation, but post-fertilisation embryonic development was not inhibited by suspended sediments. Larval survival and larval settlement were significantly reduced in high- and low-sediment treatments. No difference was found between high- and low-sediment treatments in any of the three post-spawning processes investigated, suggesting that they are susceptible to sediment concentrations which are not exceptionally high even under natural conditions (>50 mg l⁻¹). The introduction of an additional stress in the form of high levels of suspended sediments coupled with naturally high variability in recruitment may have a considerable effect on the successful supply and settlement of coral larvae to a reef. Given that many coral communities are open reproductive systems, the consequences of disturbance events are not likely to be restricted to the impact area. Recruitment to a population may be reduced significantly in the presence of high levels of suspended sediments because of effects on larval survival and settlement. Recruitment of larvae to adjacent populations may also be affected due to a decreased fertilisation success and potential increases in mortality of larvae passing through the affected site. Received: 13 August 1998 / Accepted: 22 July 1999     

Production and biomass accumulation of periphytic diatoms growing on glass slides during a 1-year cycle in a subtropical estuarine environment (Bay of Paranaguá, southern Brazil)

Production rates, chlorophyll concentrations and general composition of periphytic diatom communities growing on glass slides were studied in relation to environmental parameters during one seasonal cycle in the Bay of Paranaguá, southern Brazil. Slides were routinely submersed at 1, 2 and 3 m depth and recovered weekly for microscopic examinations, analyses of chlorophyll, cell counts and in situ photosynthetic incubations using the Winkler titration method. Water samples were also collected at surface and bottom layers for determinations of temperature, salinity, nutrients and chlorophyll in the water. The periphytic community was mainly formed by epipelic and epipsammic species, dominated by Navicula phyllepta, Cylindrotheca closterium, Navicula spp. and Amphora sp. Weekly chlorophyll a and cell accumulations on slides varied from <1–32 mg m⁻² and up to 31 × 10⁸cells m⁻², respectively. Photosynthetic rates varied from <1 to 35 mg oxygen mg chlorophyll a ⁻¹ h⁻¹, with higher values in summer. Daily production varied from 5 to 3,600 mg oxygen m⁻² day⁻¹ (<0.01–1.4 g carbon m⁻² day⁻¹). Multiple regression analysis revealed that vertical differences in light conditions and grazing pressure jointly affected the influence of temperature on the seasonal patterns of cell densities and chlorophyll concentrations according to depth. Received: 27 April 2000 / Accepted: 16 August 2000     

Sequestration of As by iron plaque on the roots of three rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivars in a low-P soil with or without P fertilizer

A pot experiment was carried out in a greenhouse to investigate the sequestration of As in iron plaques on root surface of three rice (Oryza sativa L.) cultivars. Phosphate (P) fertilization increased both plant biomass and tissue P concentrations significantly, indicating that the soils used in this study was highly P-deficient. Results from this study confirmed that low P supply improved the formation of iron plaque on rice roots. As a consequence, arsenic (As) concentrations in DCB-extracts with no P addition were significantly higher than those with P fertilization. Arsenic was highly sequestrated in iron plaque; arsenic concentration in iron was up to nearly 120 mg kg⁻¹, while arsenic concentrations in roots were just several mg kg⁻¹. Both arsenic and phosphate concentrations in iron plaque were highly positively correlated with the amounts of iron plaque (DCB-extractable Fe). Contrary to normal understanding that increasing P supply could reduced As accumulation in plants, results from the present study showed that P fertilization did not inhibit the As uptake by plants (As accumulation in aboveground), which was probably due to the fact that iron plaque formation was improved under low P conditions, thus leading to more As sequestration in the iron plaque. Thus results obtained in this study indicated that the iron plaque may inhibit the transfer of As from roots to shoots, and thus alter the P–As interaction in plant As uptake processes.     

Effects of municipal piers on the growth of juvenile fishes in the Hudson River estuary: a study across a pier edge

The growth rates of two fish species, the winter flounder Pseudopleuronectes americanus (Walbaum) (19.3 to 42.6 mm total length, TL) and the tautog Tautogaonitis (Linnaeus) (23.9 to 55.9 mm TL), were used to evaluate habitat quality under and around municipal piers in the Hudson River estuary, USA. Growth rates were measured in a series of 10 d field caging-experiments conducted at two large piers in the summers of 1996 and 1997. Cages (0.64 m²) were deployed along␣transects that stretched from underneath the piers to beyond them, encompassing the pier edge (the transitional zone between the pier interior and the outside). Growth in weight (G _w ) was determined at five locations along the transect, 40 m beneath the pier, 20 m beneath the pier, at the pier edge, 20 m beyond the pier edge, and 40 m beyond. Under piers, mean growth rates of winter flounder and tautogs were negative (xˉG _W = −0.02 d⁻¹), and rates were comparable to laboratory-starved control fishes (xˉG _W = −0.02 d⁻¹). In contrast, mean growth rates at pier edges and in open waters beyond piers were generally positive (xˉG _W ranged from −0.001 to +0.05 d⁻¹), with growth at pier edges often being more variable and less rapid than at open-water sites. Analyses of stomach contents upon retrieval of caged fishes revealed that dry weights of food were generally higher among fishes caged at open-water stations (xˉ range = 0.02 to 0.72 mg dry wt) than at pier-edge (xˉ range = 0.01 to 0.54 mg) or under-pier (xˉ range = 0.03 to 0.11 mg) stations, although it was apparent that benthic prey were available at all stations on the transect. Our results indicate poor feeding conditions among fishes caged under piers, and suboptimal foraging among fishes caged at pier edges. Inadequate growth rates can lead to higher rates of mortality, and, based on these and other earlier experiments, we conclude that under-pier environments are poor-quality habitats for some species of juvenile fishes. Received: 12 March 1998 / Accepted: 9 November 1998     

Validation of otolith growth-increment periodicity in tropical gobies

 We assessed the efficacy of tetracycline, calcein and strontium chloride for validating the periodicity of otolith growth-increments in eight species of tropical marine gobies (family Gobiidae). We compared the number of fishes in which the otoliths were successfully marked when each of these chemicals was administered by intraperitoneal injection or immersion bath at a range of doses and immersion times. All three chemicals could produce a detectable mark in the otoliths of five reef-flat gobies, Asterropteryx semipunctatus, Amblygobius bynoensis, Istigobius goldmanni, Valenciennea muralis and Amblygobius phalaena; however, tetracycline injection at 50 mg kg⁻¹ is recommended because it produced a brighter otolith mark than calcein and is cheaper and quicker to detect than strontium chloride. Calcein immersion treatment of 125 mg l⁻¹ for 24 h was the most successful treatment for two estuarine gobiids, Favonigobius reichei and Glossogobius biocellatus. No treatment produced a detectable mark in the otoliths of the coral-dwelling goby Gobiodon histrio. Adequate care in the preparation of otoliths was found to be essential for detection of the validation mark produced by tetracycline and calcein. Otolith growth-increments were deposited daily in the seven species of goby for which validation was possible. Received: 15 October 1999 / Accepted: 15 June 2000