The functions of octopine dehydrogenase and D-lactate dehydrogenase in the pedal retractor muscle of thedog whelk Nassarius coronatus (Gastropoda: Nassariidae)

The pedal retractor muscle of Nassarius coronatus, unlike most mollusc muscles, contains high activities of both octopine dehydrogenase (159 IU g^-1 wet wt muscle) and D-lactate dehydrogenase (40 IU g^-1 wet wt muscle). Pedal retractor muscles show accumulation of octopine (increase of 4.7 mol g^-1 wet wt muscle), and a fall in arginine phosphate concentration (decrease of 4.1 mol g^-1 wet wt musscle) after exercise. During recovery from exercise, octopine returns to resting levels after about 120 min, D-lactate accumulates (5.1 mol g^-1 wet wt muscle), and arginine phosphate returns to resting levels within 30 min. D-lactate (2.0 mol g^-1 wet wt muscle) accumulates during long-term exposure to air. The results show that octopine dehydrogenase and D-lactate dehydrogenase can function to catalyze the terminal step of glycolysis during muscle anoxia associated with these different physiological states.     

Larval development in the Antarctic nemertean Parborlasia corrugatus (Heteronemertea: Lineidae)

Embryonic and larval development were followed from fertilisation to settlement in the Antarctic heteronemertean Parborlasia corrugatus (McIntosh, 1876). The first cleavage occurred 10 to 15 h after fertilisation, and the second at 17 h. Larvae hatched at the gastrula stage, between 170 and 200 h post-fertilisation, and were 150 m in diameter. Early larval stages aggregated in dense groups near the surface of incubation vessels and were positively phototactic. Early pilidium larvae were recognisable 435 h post-fertilisation. They were 155×152 m in size, and possessed a complete apical tuft of cilia and a full marginal band of locomotory cilia. At this stage, the gust was visible through the body wall, and the mouth was open and was 40 m in diameter. Late pilidia, 222×193 m in size, were helmet-shaped. They had an apical tuft over 100 m long, and possessed a lobed marginal band of locomotory cilia. Pilidia were observed aggregating close to the bottom of incubation vessels 1200 to 1350 h (50 to 56 d) after fertilisation, and this was interpreted as settlement behaviour. At this stage, the apical tuft had been lost and they were highly contractile, being capable of compressing their bodies. However, neither developing juveniles within the larval envelope nor hatched juveniles were observed. Pilidia consumed the microalgae Tetraselmis suecica, Thalassiosira pseudonana and Isochrysis galbana. They also fed on particulate organic material < 1 m in size, as shown by the presence of material in the guts of larvae offered filtered extracts of algal cultures. There was some indication that larvae could use dissolved organic material, since pilidia held in seawater with organic material removed did not survive as long as those in filtered seawater or in filtered water with added amino acids. However, the only larvae to exhibit settlement behaviour in the feeding experiments were those offered Tetraselmis succica and Thalassiosira pseudonana, and these required a longer development time to reach this stage than pilidia in the standard cultures, where a mixed algal diet was offered.     

Activity patterns and predatory behavior of an intertidal nemertean from rocky shores: Prosorhochmus nelsoni (Hoplonemertea) from the Southeast Pacific

Understanding the impact of environmental stressors on predator activity is a prerequisite to understanding the underlying mechanisms shaping community structure. The nemertean Prosorhochmus nelsoni is a common predator in the mid-intertidal zone on rocky shores along the Chilean coast, where it can reach very high abundances (up to 260 ind m⁻²) in algal turfs, algal crusts, barnacle crusts, and mixed substrata. Tidal and diurnal scans revealed that the activity of P. nelsoni is primarily restricted to night and early-morning low tides and is relatively low when air temperatures are high. On average, larger worms crawled faster than smaller worms, with their maximum velocity being influenced by substratum type. Their estimated rate of predation is 0.092 prey items nemertean⁻¹ day⁻¹, just below the laboratory rate of ~0.2 amphipods nemertean⁻¹ day⁻¹ previously estimated for this species. P. nelsoni consumes a diverse spectrum of prey items (i.e., amphipods, isopods, decapods, barnacles, and dipterans) and is possibly exerting a significant influence on its prey populations. We suggest that the opportunistic predatory behavior of this intertidal predator is caused by the trade-off between immediate persistence (e.g., avoidance of desiccation) and long-term survival through successful foraging.     

Purification and partial characterisation of phenoloxidase from the colonial ascidian Botryllus schlosseri

Phenoloxidase (PO) from the colonial ascidian Botryllus schlosseri was purified using two different chromatographic strategies. A three-step purification was developed in order to maintain enzyme activity, whereas an easier purification procedure was adopted to obtain enough PO for the production of specific polyclonal antibodies. The enzyme showed optimal pH and temperature values of 7.0 to 7.5 and 35 °C, respectively, and a K _m value of 4.62 ± 0.76 mM was estimated using l-DOPA as substrate. A molecular weight of 160 kDa was determined after SDS-PAGE under non-reducing conditions. The addition of the reducing agent β-mercaptoethanol caused the disappearance of the 160 kDa band and the appearance of a new band at 80 kDa, suggesting that active PO is a dimer and the two subunits are linked by disulphide bridges. Received: 14 December 1998 / Accepted: 24 August 1999     

A blue carotenoprotein from the marine sponge Suberites domuncula: Purification and properties

A blue carotenoprotein, extracted from the marine spongia Suberites domuncula has been purified. The apoprotein has a molecular weight of about 31,000; the chromophore is not astaxanthin but it is similar to a monohydroxy, mono-expoxycarotene. The amino acid composition of the protein is briefly discussed.     

Genetic evidence for the occurrence of a cryptic species with the littoral nemerteans Lineus ruber and L. viridis (Nemertea: Anopla)

Lineus ruber and L. viridis are common intertidal heteronemerteans found on the coasts of northern Europe and the east coast of North America. The two species resemble each other morphologically and were synonymised for many years prior to being separated on the basis of larval development. In this study, specimens of L. ruber and L. viridis were collected from sites along the west and southwest coast of Britain, northern France and the east coast of North America. The external morphology and allele frequencies of isozymes of the two species were compared from all sites. The external morphology of L. ruber and L. viridis was similar but they could generally be separated by colour. Allele frequencies for up to 13 enzyme loci between sympatric populations of L. ruber and L. viridis indicated that these two species are genetically very different (Nei's genetic identity=0.090 to 0.083). Allele-frequency data also indicated the presence of a third genetic type occurring sympatrically with populations of both L. ruber and L. viridis on the coasts of Britain and France but not on the coast of North America. Fixed differences in allele frequencies between populations of the third genetic type and sympatric populations of L. ruber and L. viridis were observed across multiple loci. Genetic identity between the third genetic type and sympatric populations of L. ruber and L. viridis were extremely low (Nei's genetic identity =<0.078). Such large genetic differences between populations indicate a barrier to gene flow and reproductive isolation. The aberrant type, which exhibits sufficient morphological variation to prevent individuals being distinguished from those of both L. ruber and L. viridis, therefore represents a separate species. The low genetic identities found in intrageneric comparisons of species found in this study have been found in other studies on nemerteans. They may indicate systematic problems within these groups or other phenomena such as morphological stasis.     

Swimming behaviour of the saucer scallop Amusium balloti (Mollusca: Pectinidae)

The swimming performance of the saucer scallop Amusium balloti (Bernardi) was recorded from tests conducted in a natural environment in Shark Bay, Western Australia, in June, July and November 1984 and June and September 1985. Unlike all other scallops described in the literature, the swimming performance (both speed and distance) of A. balloti increases with size. Maximum distance swum in a single swimming event was 23.1 m, while the maximum cumulative distance swum (four swimming events) was 30.8 m. Swimming speeds for larger scallops were generally between 0.8 and 1.0 m s^-1 (1.6 and 2.0 knots), with a maximum speed of 1.6 m s^-1 (3.1 knots). Variations in swimming performance and response times with size and season are probably the major cause of variations in the scallop's vulnerability to fishing gear.     

Respiratory metabolism of crabs from marine and estuarine habitats: an interspecific comparison

The gill-surface area and R-T (metabolic rate-temperature) response during aquatic and aerial respiration of three different species of crabs, the subtidal Scylla serrata, the intertidal Sesarma quadratum, and the supratidal Ocypoda platytarsis, were compared. Scylla serrata has the highest, Sesarma quadratum the second highest, and O.platytarsis the lowest gill-surface area. Oxygen-consumption values in water under different ambient partial pressures of oxygen indicate that all three species display equally efficient aquatic respiration despite variation in gill area and number. There is no coherent R-T response either inter- or intra-specifically. However, the ability to extract oxygen from the surrounding medium (respiratory efficiency) conditions the R-T response in the different species. The R-T trends in aquatic respiration of Scylla serrata and aerial respiration of O.platytarsis reveal a normal response to temperature. The proportion of aquaticoxygen uptake to total respiration in all sizes in both Scylla serrata and Sesarma quadratum remains more or less constant (between 88 and 92%), whereas in the supratidal O.platytarsis, this proportion decreases with increasing weight (from 88 to 64 %), indicating a progressive loss of gill respiration to total respiration. All three species are metabolically equally efficient; what one lacks in respiratory surface it compensates by respiratory efficiency.     

Extracellular polymeric substances (EPS) from cyanobacterial mats: characterisation and isolation method optimisation

Extracellular polymeric substances (EPS) play an important role in bacterial mat formation and sediment stabilisation of coastal zones. The analysis of these secretion products on a molecular level is a prerequisite to understand their formation mechanisms and ecological function in microbial consortia. The present study focuses on the optimisation of EPS isolation and characterisation from cohesive cyanobacterial mats. Extracted EPS were analysed for quantity, content of total organic carbon and nitrogen, and bulk contents of neutral sugars, uronic acids, and proteins. These criteria are strongly influenced by the extraction conditions applied and therefore, the effects of different extraction media (NaCl or artificial seawater), addition of EDTA, centrifugal force, and purification via repeated ethanol precipitation on extraction success were determined. From this an optimised extraction procedure for EPS resulted. When using fresh mat samples, the yield of EPS amounted to 3.3 ± 0.8 mg g⁻¹ mat (dw). The isolated EPS were composed of nearly equal amounts of proteins and uronic acids (12.7 ± 1.5 and 11.8 ± 1.1%, respectively) and the bulk content of neutral sugars was 30.5 ± 2.6%. High contents of TOC and TN indicated relatively pure EPS and only a low contribution of inorganic compounds. Furthermore, low contents of 2-keto-3-deoxyoctonate and the small protein/polysaccharide-ratio in the EPS extracted by our method, signified low contaminations by intracellular polymers and hence a low rupture of cells. Our method provides an useful tool to evaluate further investigations of the composition, characteristics and properties of EPS on a sound basis.     

Seasonal variation in the properties of muscle mitochondria from the tropical scallop Euvola (Pecten) ziczac

In scallops, gametogenesis leads to considerable transfer of energetic reserves from the adductor muscle to the gonads. During an annual cycle, the scallops are exposed to changes in temperature and food availability. As these changes may affect muscle metabolic capacities, we examined whether the properties of the mitochondria in the phasic adductor muscle were modified during the annual cycle of the scallop Euvola (Pecten) ziczac (L. 1758). During our study, temperature and chlorophyll a levels generally showed an inverse relationship: high temperatures and low chlorophyll a levels occurred from mid-April to early June. Lower temperatures and higher chlorophyll a levels were found from January to late March and from mid-June to mid-September. Throughout the annual cycle, the substrate preferences and the pH sensitivity of the isolated muscle mitochondria changed little, whereas the maximal oxidative capacities and respiratory control ratios (RCR) varied considerably. Consistently, the maximal capacities for substrate oxidation were 30 to 80% lower in mitochondria isolated in May than at other times in the year. The RCR values of mitochondrial oxidation of glutamate, glutamine and succinate varied throughout the year with lower values characterizing the mitochondria from scallops harvested in May and in certain cases in August. In May, adductor muscles had lower protein levels than at other times. These data suggest that the requirements of gametogenesis, coupled with␣the high temperatures and low food availability occurring during April and May, led to a mobilization of muscle proteins which concomitantly decreased the oxidative capacity of isolated mitochondria. Received: 29 November 1996 / Accepted: 5 December 1996     

Optimizing the design of giant scallop (Placopecten magellanicus) spat collectors: field experiments

Field experiments were carried out in 1991 and 1992 in the Gulf of St. Lawrence (Québec, Canada) to examine the influence of various structural components of spat collectors on recruitment of the giant scallop (Placopecten magellanicus) and associated species. Components considered were monofilament diameter, heterogeneity (presence of knots), mesh size of external bags combined with monofilament density, and spatial arrangement of monofilament inside collectors. P. magellanicus spat were more abundant in collectors made with bags of intermediate mesh size (0.4x0.5 cm) containing low densities (2030 cm²) of monofilament than in other types of collectors. significant variations in spat numbers were, however, observed with other taxa such as Mytilidae, Hiatella arctica, Cerastoderma pinnulatum, Mya arenaria, Anomia simplex, Tellina sp., Veneridae, Dacrydium vitreum, Mactridae, Buccinidae, Littorina sp., Ostracoda and Foraminifera. Abundance of scallop spat did not vary significantly with spatial arrangement, diameter and heterogeneity of monofilament.Contribution to the program of GIROQ (Groupe Interuniversitaire de Recherches Océanographiques du Québec) and REPERE (Recherche sur le Pétoncle à des fins d'Élevage et de Repeuplement).     

Optimizing the design of giant scallop (Placopecten magellanicus) spat collectors: flume experiments

Flume experiments were carried out in 1993 with specially designed collectors and inert particles to optimize the effect of various structural components of scallop spat collectors. The structural components considered were monofilament diameter and density, and mesh size of external bags. Results showed that abundance of particles trapped in silicone-treated collectors did not vary significantly with monofilament diameter. The heterogeneous arrangement of monofilament also proved to be an important factor. Combinations of monofilament of three different diameters (0.17, 0.55 and 0.90 mm), the thinnest being the upstream one, collected at least twice as many particles than all other single one-sized monofilament collectors. Particle abundance decreased with decreasing density of monofilament, and the densest arrangement of monofilament captured over four times more particles than the least dense arrangement. Collectors with an external bag of intermediate mesh size (0.4×0.5 cm) were the most efficient at collecting particles. The effect of flow speed was also investigated. Particles were trapped in significantly greater numbers at flow velocities of 5 cm s^-1 than at lower or higher velocities. These results are consistent with results of field experiments carried out to optimize the design of scallop collectors. Enhancement of spat collection is possible by modifying collectors. Results from both field and flume experiments confirm the hypothesis that larvae of the species collected in the field are passive at settlement at the scale of the collectors.Contribution to the program of GIROQ (Groupe Interuniversitaire de Recherches Océanographiques du Québec), and OPEN (Ocean Production Enhancement Network)     

Phylogeography of the marine interstitial nemertean <Emphasis Type="Italic">Ototyphlonemertes parmula</Emphasis> (Nemertea,Hoplonemertea) reveals cryptic diversity and high dispersal potential

We conducted a phylogeographic study of the meiofaunal nemertean Ototyphlonemertes parmula, an apparent species complex from the littoral zone of coarse-grained beaches, using a 494-bp fragment of the mitochondrial cytochrome oxidase 3 gene (cox3). Six populations from the Gulf and Atlantic coasts of Florida, two from New England, and one from the Caribbean were sampled in March and August 2005. Three major lineages were identified, separated by cox3 sequence divergence of 16–18%, with partially overlapping ranges. Tests for hybridization using ISSR markers detected nuclear gene exchange within but not between the major mitochondrial lineages, indicating the presence of cryptic species. One lineage dominating the Atlantic coast of Florida shows no evidence of geographic structuring. Another lineage shows a phylogenetic break between the Atlantic and Gulf coasts, suggesting that unsuitable habitat may act as a barrier to dispersal. Long-distance migration is evidenced by shared haplotypes between Florida and the eastern Caribbean. Overall, the widespread distribution of individual haplotypes and lack of structuring within geographic regions contrast with O. parmula’s strongly sediment-bound lifestyle. We speculate that dispersal of adults by storms and/or sediment transport may be more important than few and potentially short-lived planktonic larvae to explain geographic diversity in O. parmula and may be important for meiofauna in general.     

Delayed growth of mussel (Mytilus edulis) and scallop (Pecten maximus) veligers at low temperatures

Veliger larvae of Mytilus edulis (L.) from Menai Straits, North Wales, were maintained for up to 2 mo during 1981 at 5°C and then grown on to metamorphosis at 17°C. Larvae so treated showed similar low mortality and equivalent spat production to control larvae. Growth rate at 17°C was less in treated larvae than in controls, but treated larvae grew a little during the period at low temperature. Larvae of Pecten maximus (L.) from the Irish Sea suffered high mortality at low temperature but larvae surviving 2 wk at 8°C could be grown on to spat at 17°C. The longevity of M. edulis larvae is discussed in relation to the genetic homogeneity of adult populations around the UK.     

Heavy metals uptake from aqueous solutions using marine algae (Colpomenia sinuosa): kinetics and isotherms

The adsorption of copper, zinc, cobalt, lead and cadmium ions onto Colpomenia sinuosa was studied as a function of contact time, initial metal ion concentration and initial pH. In addition, desorption studies were performed. Characterisation of this adsorbent was also confirmed by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) analysis. Batch adsorption experimental data were analysed using Langmuir, Freundlich and Dubinin–Raduschkevich (D–R) adsorption isotherms. The results indicated that the biosorption equilibrium was well described by both the Freudlich and D–R isotherms. Moreover, sorption kinetics was performed and it was observed that equilibrium was reached in<60 min, which could be described by the pseudo-second-order kinetic model for all heavy metals. The sorption of heavy metals onto the biomass was largely dependent on the initial solution pH. The elution efficiency for heavy metal ions desorption from C. sinuosa was determined for 0.1 M HCl, 1.0 M HCl and 1.0 M HNO₃. Desorption efficiency and also adsorption capacity were highest for Pb(II). The results indicate that C. sinuosa has great potential for the removal of heavy metals in an ecofriendly process.     

Multivariate characterisation of the habitats of seven species of Malayan mudskippers (Gobiidae: Oxudercinae)

Mudskippers (Gobiidae: Oxudercinae) are amphibious gobies. The occurrence and habitats of seven sympatric species of Malayan mudskippers were recorded: Boleophthalmus boddarti, Boleophthalmus pectinirostris, Periophthalmodon schlosseri, Periophthalmus chrysospilos, Periophthalmus gracilis, Periophthalmus variabilis and Scartelaos histophorus. A total of 14 surveys were made at six localities along 120 km of the west coast of the Malay Peninsula in August 1996 and September 2006. A multivariate set of ordinal parameters were used to measure the quantity of environmental water in mudskippers’ habitats, and three guilds of species were discriminated. The resulting pattern mirrored the adaptive scenario depicted by physiological and anatomical studies of mudskippers’ terrestriality, although in one case a terminal taxon was less terrestrial than other more basal ones. Differentiation of trophic niches may also play a role in habitat selection by the herbivorous Boleophthalmus spp., which were only found in unvegetated, illuminated areas and on wet mud at low tide. With the more terrestrial guild, a significant negative co-occurrence suggested direct interspecific competition. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Incorporation of dietary sterols by the sea scallop Placopecten magellanicus (Gmelin) fed on microalgae

The anatomical distributions of sterols and the incorporation of dietary phytosterols into different organs were studied in two populations of sea scallops, Placopecten magellanicus Gmelin, collected in 1989 from Georges Bank (Nova Scotia) and St. Pierre Bank (Newfoundland), respectively. In contrast to the well-established organ-specific lipid classes and fatty-acid compositions usually found in marine animals, the major organs of wild sea scallops (adductor muscle, digestive gland, gonads, gills and mantle) had the same sterol compositions. In order to know if anisomyarian bivalves require a uniform anatomical distribution of sterols, wild scallops were subjected to a microalgal diet containing high concentrations of brassicasterol, -sitosterol and cholesterol. The sterol composition of the scallop adductor muscle was not changed by 6 wk of feeding on the experimental diet. In contrast, the proportions of brassicasterol, -sitosterol and cholesterol in the digestive gland, and of brassicasterol and cholesterol in the male gonad, increased significantly (p<0.05). These results showed that the typical even anatomical distribution of sterols of bivalves can be disrupted by a drastic change in diet and is therefore not subject to strict internal regulation. Furthermore, the P. magellanicus results indicate that, although sea scallops may be capable of sterol biosynthesis, the incorporation of unmodified dietary phytosterols plays an influential role in establishing their sterol composition.ORNL is managed by Martin Marietta Energy Systems for the US Department of Energy, under contract DE-ACOS-8-10R21400     

Molecular and ecological characterisation of plant populations from limed and metal-contaminated sites in Northern Ontario (Canada): ISSR analysis of white birch (Betula papyrifera) populations

The Greater Sudbury region in Northern Ontario is known as one of Canada's most ecologically disturbed regions because of the effects of heavy metal pollution. The main objectives of the present study were to assess the effects of soil liming of sites contaminated with metals on species richness and abundance, forest health in general, and to determine the level of genetic variability in white birch populations from Northern Ontario. Shannon-Wiener diversity index and tree species richness values were higher in populations from limed and control sites compared to the unlimed areas. A significant improvement in forest population health (measured using a scale of 1 to 10) in limed sites over the unlimed areas was observed. Key results revealed no significant difference for metal content in white birch (Betula papyrifera) leaves from limed compared to unlimed sites. But higher levels of Al, Ca, Mg, Mn, Ni, Sr, and Zn in leaves compared to the bioavailable amount in soil were observed. The levels of genetic variability in white birch populations were moderate to high, ranging from 30% to 79% of polymorphic loci. A high level of genetic variability such as observed in the present study is usually associated with long term sustainability in plant populations. No association was found between metal accumulation in soil or plants and the levels of genetic variation.     

近平滑假丝酵母NAD(H)依赖型次级醇脱氢酶的分离纯化及酶学性质

从近平滑假丝酵母(Candida parapsilosis CCTCC 203011)中分离得到了NAD(H)依赖型的次级醇脱氢酶.粗酶经乙醇沉淀、Blue Sepharose Fast Flow亲和层析、DEAE Sepharose离子交换层析后在SDS-PAGE上显示为单一条带,其酶蛋白的亚基分子量(Mr)为37.5×103.该酶还原反应的最适pH为6.0,最适温度为45℃,是一个含Zn2 金属酶.该脱氢酶具有较高的底物的选择性,对2-酮具有较高的还原活力;同时,也能作用于醇类发生氧化反应,对次级醇的氧化活力最高.以β-羟基苯乙酮为底物,用纯化得到的酶催化反应后,还原产物为(R)-苯基乙二醇,因此该酶为(R)-专一性次级醇脱氢酶,是一种生产(R)-苯基乙二醇的有效的生物催化剂.经LC-MASS-MASS分析得到了酶蛋白中3个肽段的氨基酸序列,通过比对发现该酶与NCBI编号为BAA24528的次级醇脱氢酶具有较高的同源性.图4表3参17     

An apparatus for measuring photosynthesis and respiration of intact large marine algae and comparison of results with those from experiments with tissue segments

An apparatus is described for incubating large marine algae in light and dark chambers on the sea floor, while providing vigorous circulation of water prefiltered to remove phytoplankton. Successful use of the equiment in over 500 h of incubations of Laminaria longicruris has shown that earlier methods in which strips of tissue are incubated in bottles greatly overestimates dark respiration rates and increases experimental error. The effect of these findings on long-term productivity estimates is discussed.