The effect of environmental factors on the absorption of amino acids by isolated gill tissue of the bivalve, Mya arenaria (L.)

The absorption of amino acids by isolated gill tissue of Mya arenaria (L.) was examined under conditions in which the environmental temperature, salinity, and pH were varied experimentally. A decrease in salinity resulted in a fall in amino-acid uptake. The V_max of L-histidine uptake increased with temperature, the Q₁₀ for the range 5° to 15 °C being 2.64. No significant effect of pH on amino-acid absorption was observed over the pH range 6.2–8.8. These results emphasize the need clearly to specify the experimental conditions when substrate uptake values are used to calculate the contribution made by any extra-alimentary absorption route to the nutritional economy of an animal.     

Burrowing behaviour of the softshell clam (Mya arenaria) following erosion and transport

Erosion and transport of juvenile individuals may alter the distribution pattern of intertidal bivalves. The burrowing success of recently transported juvenile softshell clams (Mya arenaria) was studied in a laboratory flume under a wide range of hydrosedimentary environments. Juvenile individuals (5-20 mm) were observed under a simulated 30 min slack tide before initiating the flow for a period of 60 min. Five different free-stream velocities (0, 3, 5, 10 and 24 cm s^− 1) and four sediment types (mud, sandy-mud, sand and gravel) were used. The mean proportion of juvenile clams that initiated (MPI) or completed (MPC) a burial decreased with increasing shell length. Erosion from the sediment was more important in large juveniles suggesting that large juveniles may have more difficulty successfully relocating once transported. The MPI increased with increasing flow speed in experimental runs held at speed < 24 cm s^− 1. This was observed in all sediment types. Most individuals were unable to burrow at 24 cm s^− 1 because they got eroded. The MPC also increased with increasing flow speed in mud, sandy-mud and sand. The MPC's response to flow was more complex in gravel because of a shell length × flow speed interaction effect. Our observations suggest that water movement may induce the burrowing behaviour of recently eroded juvenile clams. Results are discussed in an ecological and aquacultural context.     

Behavioral plasticity of the soft-shell clam, Mya arenaria (L.), in the presence of predators increases survival in the field

Soft-shell clams, Mya arenaria, are sessile, suspension-feeding bivalves that are preyed upon by the exotic green crab, Carcinus maenas. Clams evade crab consumers by burrowing deeper into the sediment after perceiving a threat from a nearby predator. The purpose of this study was to determine the types of signals that M. arenaria use to detect predators and the types of behaviors clams use to avoid being eaten. In a field study, clams increased their burial depth in the presence of green crab predators consuming conspecifics that were caged nearby, and also increased burial depth after artificial tactile stimulation in the laboratory assay. These results indicate that clams can use chemical and mechanical cues to detect potential predatory threats. We performed a field study to examine the difference in survivability of clams that had burrowed deeper into the sediment in response to predators vs. control clams that were burrowed less deeply. Significantly higher survival rates were observed in clams that had initially burrowed more deeply, suggesting that increasing burial depth is a valid predator avoidance strategy. Some bivalves also alter their pumping rates in the presence of predators, making them less apparent and providing more structural defense by covering soft tissue, and we measured pumping time of soft-shell clams in the presence and absence of predators, when burrowing was not an option for escape. Soft-shell clams did not alter their pumping time in the presence of green crab predators, possibly because they employ a burrowing method called “hydraulic” or “jet-propelled” burrowing, where it is necessary for the clam to pump in order to burrow. Chemical signals and tactile cues instigated behavioral changes in M. arenaria, and this change in behavior (increasing burial depth) increased clam survival in the field.     

Effects of current speed, shell length and type of sediment on the erosion and transport of juvenile softshell clams (Mya arenaria)

The erosion and transport of juvenile softshell clams (Mya arenaria) was studied in a laboratory flume in relation to free-stream velocity (0, 7, 16, 29 and 35 cm s^− 1), shell length (0-5, 5-10, 10-15, 15-20 mm) and type of sediment (mud, sandy-mud, sand and gravel). Our results showed that these factors interact together on the erosion of clams from the sediment. Juveniles were eroded in great numbers in sand while mud retained them more easily. Bedload transport was initiated at speeds of 16 cm s^− 1. Most of the clams were eroded in sandy sediments at speeds of 29 and 35 cm s^− 1. The smallest individuals were highly vulnerable to erosion compared to the other size classes studied. A results-based model using the logistic regression statistics was proposed. This allowed the estimation of erosion probabilities for a given hydrosedimentary environment. A field validation of the model was then carried out. Field results confirmed the importance of free-stream velocity, shell length and type of sediment on the erosion rate of clams. The differences observed between predicted and field results suggest that the model underestimated the erosion rate in the field. Results are discussed in the context of hydrosedimentary environments found off the eastern coast of Canada.     

Distances of dispersal of juvenile bivalves (Mya arenaria (Linnaeus), Mercenaria mercenaria (Linnaeus), Gemma gemma (Totten))

Although it is recognized that many species of benthic invertebrates continue to disperse after settlement, particularly in soft-bottom habitats, the scale over which movements of juveniles occur is not well known. This study combined laboratory flume experiments assessing the effects of clam size, species, and water velocity on rates and distances of dispersal of three species of juvenile bivalves with field measurements of loss rates and distances of dispersal of transplanted bivalves in the Navesink River estuary in New Jersey, USA. Dispersal distances measured in the laboratory ranged from an average of 1.6 to 40 cm h^− 1 depending on clam size, species, and flow speed. Distances and likelihood of dispersal were generally greater for Mya arenaria than for Mercenaria mercenaria or Gemma gemma, although differences between species were not consistent. As predicted, smaller (1.3 mm) M. arenaria tended to disperse more than larger (3.7 mm) ones, although no significant differences were detected between two sizes (1.8 and 3.4 mm) of M. mercenaria. The similarity of the erosion thresholds of dead clams across sizes and species suggests that burrowing behaviour plays an important role in determining variation in dispersal due to clam size and species. In the field, densities of clams (M.arenaria and M.mercenaria) were reduced to half of that in controls after 3.5-5 h, indicating high levels of dispersal and/or mortality. Some individuals were recovered up to 50 cm away from their initial locations. Overall, our results suggest that dispersal distances of these three species due to bedload transport are likely to be on the order of centimeters per hour. Although these dispersal distances are small, such movements are likely to occur frequently due to tidal currents and, consequently, may have profound impacts on patterns of abundance and distribution.     

Active transport of alanine by the neutral amino-acid exchanger ASCT1

ASCT1 protein is a member of the glutamate transporter superfamily, which shows system ASC selectivity and properties and has been characterized as a Na+-dependent neutral amino-acid exchanger. Here, by using ASCT1-expressing oocytes, the uptake of alanine and glutamate was measured to investigate ASCT1's ability to mediate a concentrative transport of alanine, ASCT1's sodium dependence, and the influence of pH on the mutual inhibition between alanine and glutamate. Alanine uptake was measured after 30 min incubation. Kinetic analysis of the Na+ dependence of alanine uptake showed an apparent K0.5 (affinity constant) value for Na+ of 23.1 +/- 4.3 mM (mean +/- SE). Concentration dependence of alanine uptake was tested at 100 and 1 mM Na+, with apparent K0.5 values of 0.16 +/- 0.04 and 1.8 +/- 0.4 mM, respectively, at pH 7.5, and 0.21 +/- 0.06 and 1.9 +/- 0.3 mM at pH 6. Vmax was not modified between 100 and 1 mM Na+ at either pH. ASCT1 actively transports alanine and accumulates it in the cytosol even when the Na+ concentration in the medium was as low as 1-3 mM. 22Na uptake studies revealed that Na+ transport was stimulated by the presence of alanine in the medium. Our results demonstrate that ASCT1 is able to mediate a concentrative transport of alanine, which is Na+-dependent but not coupled to the Na+ gradient.     

The mechanism of calcium transport in mitochondria isolated from the marine mussel, Mytilus edulis (L.)

Isolated mussel mitochondria produced a less pronounced transient stimulation of respiration upon the addition of Ca²⁺ in a reaction medium containing Pi and a slower rate of Ca²⁺ transport compared to rat liver mitochondria. The initial rates of Ca²⁺ transport in the absence of Pi were more similar and both types of mitochondria possessed a sigmoidal relationship between the initial rate of Ca²⁺ transport and the free Ca²⁺ concentration $\text{(‘Km’ ? 5μM)}$. Ruthenium red produced an equal maximal inhibition of the initial rate of Ca²⁺ transport in both types of mitochondria but mussel mitochondria were rather more resistant to the inhibitor. The major difference found was that approximately 15 nmoles La³⁺ mg protein^?1 was required to produce maximal inhibition of the initial rate of Ca²⁺ transport in mussel mitochondria compared to approximately 1.0 nmole La³⁺ mg protein^?1 in rat liver mitochondria. It is concluded that mussel mitochondria possess a comparable Ca²⁺ transporter to vertebrate mitochondria and possible reasons for resistance to La³⁺ are discussed.     

Calorific values of the soft parts of the tellinid bivalve Macoma balthica (L.) as determined by two methods

Calorific values of the ash-free dried soft parts of Macoma balthica (L.), sampled monthly during 1 year at a tidal flat station in the Dutch Wadden Sea, have been estimated by two methods. The indirect method started from an analysis of the biochemical composition and used conversion factors for the various components to calculate the calorific value of the total tissue. The direct method employed a micro-bomb calorimeter and a furnace for separate ash determinations.The indirect method yielded estimates that were consistently higher than those from the direct one. Two sources, explaining most of the difference, could be traced, viz. incomplete drying of the samples used in the calorimeter and a too high conversion factor for weight to calories applied conventionally for lipids. This conversion factor was found to depend on the method of extraction of the lipids. but was always significantly below the commonly used 9.3 to 9.7 cal · mg^?1.The best estimates for the annual mean of the calorific value at complete combustion of the ash-free and well-dried soft parts of Macoma were 5.47 for the direct and 5.59 cal·mg^?1 ash-free dry wt for the indirect method. Both values are higher than estimates for Macoma published earlier by various authors. Calorific values at physiological oxidation amount to only about 4.6 cal·mg^?1 or less, due to incomplete protein oxidation.For this reason, it is concluded that biologically significant calorimetry cannot dispense with biochemical analysis. On the other hand, calculation of calorific values from biochemical composition should be accompanied by some calorimetry, as the conversion factor for the lipid fraction, at least, may deviate significantly from commonly applied values.     

Amino acid transport in the gill epithelium of a marine bivalve

1. Amino acid transport across the intestine of bivalve molluscs is reviewed. 2. Transport of alanine or taurine is dependent on the sodium gradient across the intestine. 3. The time course of the uptake of alanine into the brush border membrane vesicles is also sodium dependent.     

Effects of hypoxia and body size on the oxygen consumption of the bivalve Arctica islandica (L.)

The relationship between oxygen consumption and body weight has been examined in Arctica islandica (L.).Large Arctica exhibit a high degree of respiratory independence under hypoxic conditions; like other species studied, respiratory independence increases markedly with increasing body size. Respiratory independence can also be modified by temperature and physiological condition and it is concluded that the division of species per se into oxygen regulators and oxygen conformers is not merited. These terms describe only the extremes of what is a variable capacity to maintain respiratory independence during hypoxia.     

Relative importance of predation and intraspecific competition in regulating growth and survival of juveniles of the soft-shell clam, Mya arenaria L., at several spatial scales

Predation appears to be the single most important biotic factor regulating populations of bivalves in estuarine and coastal soft sediments. However, the relative roles of predation and intraspecific competition are rarely investigated simultaneously over different spatial scales, making generalities about these mechanisms difficult. Using juveniles of the soft-shell clam, Mya arenaria (initial mean shell length [SL] ± 95% CI = 12.4 ± 0.13 mm), I tested the interactive effects of predator deterrence and intraspecific density (660 vs. 1320 individuals m^− 2) on growth and survival responses over a 185-day period from May to November 2003 at spatial scales that spanned four orders of magnitude: embayments, sites within embayments, tidal gradients, and blocks that were 10,000's, 1000's, 100's, and 5 m apart, respectively. Replicate field experiments were conducted from May to November 2003 at the upper and lower tidal heights at each of two intertidal mud flats (sites) within each of two embayments (Passamaquoddy Bay [PB] and Cobscook Bay [CB]) in eastern Maine.Mean survival, relative growth, and the abundance of wild recruits each varied significantly over all spatial scales. Predation was the most important factor affecting clam survival, explaining 45% of the total variability, whereas embayment, sites within embayments, tidal gradient, and intraspecific density collectively accounted for less than 10% of the variation. At all four intertidal sites, clam survival in experimental units designed to deter predators averaged 72%, but the degree of enhancement varied between embayments (PB = 61%; CB = 267%). Average survival rate was higher (by 12-16%), but growth was slower (by ca. 50%) in upper vs. lower intertidal blocks; however, the patterns differed for both variables between sites within each embayment. The effect of increasing intraspecific clam density was to lower survival by ca. 17% (56% [660 m^− 2] vs. 48% [1320 m^− 2]) in both embayments, but growth was unaffected. Overall, clams doubled in SL, although mean relative growth was 15% greater in CB than PB. Tidal gradient, sites within embayments, and blocks were the three most important factors explaining 35%, 19%, and 22% of total variation in relative clam growth, respectively. In Maine and the northeast US, juveniles of Mya reach their highest abundance above mean low tide levels. Experimental evidence presented here suggests that differential predation along the tidal gradient is the dominant factor controlling clam abundance and distribution patterns in the intertidal zone.     

Hoarding by honeybees (Apis mellifera L.)

The food hoarding by groups of fifty bees kept in small cages and provided with sugar syrup was studied. Less food was stored in a new comb than in an old one, whether the old comb had been used for storing food or rearing brood, and there was less in drone than in worker combs. The presence of light, larvae and the odour of honey discouraged storage of syrup, but the presence of a queen encouraged it. The amount stored also varied with the environmental temperature, the age of the bees concerned and with their previous physiological and behavioural experience including food deprivation and length of confinement. Increased food in honeystomachs sometimes compensated for less stored in combs.     

Chemoreception by the asteroid Cross aster papposus (L.)

Chemoreception abilities were demonstrated in the solasterid asteroid Crossaster papposus (L.) from a level substratum in the current-swept Crouch Estuary, England. C. papposus showed wide variability of positive and negative responses to crude extracts of prey species and (unattractive) conspecifics. C. papposus has an olfactory (distance chemoreception) ability to whole, live potential prey and conspecifics. This was confirmed by contact chemoreception responses, which are stronger, to the same stimuli. The olfaction tests in the Y-maze yielded variable results in which the initial (rheopositive or rheonegative) responses demonstrated that conspecifics and prey species were avoided yet subsequent arm choices of rheopositive individuals showed significant selection for prey over controls and no avoidance of conspecifics. The arm choices for prey relate to a priority of orientation to food and feeding over intraspecific avoidance in hungry asteroids. Intraspecific avoidance functions more in dispersion than prevention of cannibalism and is less important than feeding to hungry asteroids. Interpretation of chemically-mediated responses of asteroids requires appreciation of their innate behavioural variability which has led to confusion in the literature. This is the first experimental study of contact and distance chemoreception among solasterids. Their chemosensory abilities have previously been contradictory which may be due to intraspecific avoidance in satiated individuals of some species.     

Comparative study of lignins isolated from Alfa grass (Stipa tenacissima L.)

Soda lignin, dioxane lignin and milled lignin were isolated from Alfa grass (Stipatenacissima L.). The physico-chemical characterization of three different lignins: one industrial lignin precipitated from soda spent liquor and two lignin preparations isolated under laboratory conditions from Alfa grass (also know as Esparto grass) was performed. The structures of lignins were studied by three non-destructive (FT-IR, solid state ¹³C NMR and UV/visible spectroscopy) and two destructive (nitrobenzene oxidation and thermogravimetric analysis) methods. Elemental analysis and the methoxyl content determination were performed in order to determine the C₉ formulae for the studied lignins. The total antioxidant capacity of the studied lignins has been determined and compared to commercial antioxidants commonly used in thermoplastic industry.     

In vitro excystment of the metacercaria of Maritrema arenaria (Digenea: Microphallidae)

Metacercarial cysts of Mantrema arenaria were subjected to a solution containing trypsin and bile salts at 41°C. This treatment induced intense metacercarial activity and after 15 min metacercariae burst through their cyst walls and emerged. Electron microscopy demonstrated that during the process of excystment the inner layer of the cyst wall changed from a compact to a loose fibrous state. Experiments showed that only cysts containing viable metacercariae underwent this change whereas cysts which had been forcibly vacated before treatment did not. This indicated that the structural change of the inner layer of the cyst wall could not be attributed to the excystment medium. Also there was much less acid phosphatase activity in and on the surface of newly excysted metacercariae compared with encapsulated specimens. It was concluded that the excystment medium induced physical activity in, and the release of enzymic material by, the metacercariae. Together these activities rendered the cyst wall soft and susceptible to rupture by physical pressure.     

Changes in shell and soft tissue growth, tissue composition, and survival of quahogs, Mercenaria mercenaria, and softshell clams, Mya arenaria, in response to eutrophic-driven changes in food supply and habitat

Eutrophic-driven changes in the composition of near-bottom seston and surface sediment potentially affect food resources and habitat of commercially important bivalves like quahogs, Mercenaria mercenaria, and softshell clams, Mya arenaria. To define how land-derived nitrogen loads and resulting eutrophication affect bivalves, we compared estuarine features to growth and survival of clams across estuaries receiving different N loads. The major effects of nitrogen enrichment on near-bottom seston and surface sediment were to (1) increase microalgal concentrations and reduce carbon to nitrogen ratios, increasing quantity and quality of available foods, and (2) reduce oxygen content in sediments, potentially reducing habitat quality. Shell growth of juvenile and native clams increased with increasing food supply, driven by N enrichment. Growth of soft tissue followed growth of shell, and %N content of soft tissue increased across N loads, providing direct evidence of a link between N loads and growth responses in clams. In some locations, low salinity limited growth and low oxygen concentrations may have reduced survival. Despite these factors, our data indicate the major effect of N enrichment on clams was increased secondary production in terms of shell and soft tissue growth.     

Silicon decreases chloride transport in rice (Oryza sativa L.) in saline conditions

Silicon can alleviate salt damage to plants, although the mechanism(s) still remains to be elucidated. In this paper, we report the effect of silicon on chloride transport in rice (Oryza sativa L.) seedlings in saline conditions. In the absence of salinity, silicon enhanced the growth of shoots, but not roots in three cultivars (cv. GR4, IR36, and CSR10). Salinity reduced the growth of both shoots and roots in all three genotypes. In saline conditions, addition of silicon to the culture solution again improved the growth of shoots, but not of roots. Under these saline conditions, the concentrations of chloride in the shoot were markedly decreased by adding silicon and the ratio of K⁺/Cl⁻ was significantly increased, while the concentration of chloride in the roots was unchanged. The decrease in chloride concentration in the shoot was correlated with the decrease in transpirational bypass flow in rice, as shown by the transport of the apoplastic tracer trisodium-8-hydroxy-1,3,6-pyrenetrisulphonic acid (PTS). Addition of silicon increased the net photosynthetic rate, stomata conductance, and transpiration of salt-stressed plants in cv. IR36, indicating that the reduction of chloride (and sodium) uptake by silicon was not through a reduction in transpiration rate. Silicon addition also increased the instantaneous water use efficiency of salt-stressed plants, while it did not change the relative growth rate of shoots. The results suggest that silicon addition decreased transpirational bypass flow in the roots, and therefore decreased the transport of chloride to the shoot.     

A chloride requirement for Na+-dependent amino-acid transport by brush border membrane vesicles isolated from the intestine of a mediterranean teleost (Boops salpa)

The uptake of d-glucose, 2-aminoisobutyric acid and glycine was studied with intestinal brush border membrane vesicles of a marine herbivorous fish: Boops salpa. The uptake of these three substances is stimulated by an Na⁺ electrochemical gradient ($\text{C}{}_{\mathrm{<mtext>out</mtext>}}\text{C}{}_{\mathrm{<mtext>in</mtext>}}$). For glucose, an increase of the electrical membrane potential generated by a concentration gradient of the liposoluble anion, SCN^?, increases the Na⁺-dependent transport. This responsiveness to the membrane potential was confirmed by valinomycin. Differently from glucose, uptake of glycine and 2-aminoisobutyric acid requires, besides the Na⁺ gradient, the presence of Cl^? on the external side of the vesicles. In the absence of Cl^?, amino acid uptake is not stimulated by the Na⁺ gradient and is not influenced by an electrical membrane potential generated by SCN^? gradient ($\text{C}{}_{\mathrm{<mtext>out</mtext>}}\text{>C}{}_{\mathrm{<mtext>in</mtext>}}$) or by a K⁺ diffusion potential ($\text{C}{}_{\mathrm{<mtext>in</mtext>}}\text{>C}{}_{\mathrm{<mtext>out</mtext>}}$). This Cl^? requirement differs from the Na⁺ requirement, since a Cl^? gradient ($\text{C}{}_{\mathrm{<mtext>out</mtext>}}\text{>C}{}_{\mathrm{<mtext>in</mtext>}}$) does not result in an accumulation of glycine or 2-aminoisobutyric acid similar to that produced by an Na⁺ gradient.     

Dormancy of Medicago marina (L.) seed

Medicago marina (L.) is a Mediterranean species whose seeds show strong dormancy that prevents germination. We used an integrated approach of physiological analyses and proteomics to investigate the mechanisms that control M. marina dormancy/germination and that underlie stress tolerance. First, we evaluated the effects on dormancy breaking of the following treatments: mechanical scarification, freezing at −20 °C, storage for 4 months and heating at 100 °C for 1 h. Mechanical scarification and freezing were the most effective treatments in overcoming dormancy. The role of abscisic acid (ABA) in M. marina dormancy was studied by ELISA immuno-enzymatic assay. The ABA content of germinated and non-germinated mature (control) and treated seeds was determined. The level of ABA was higher in treated seeds than in control seeds; the most significant increase occurred in the heated seeds. A comparison of the ABA level in the germinated, control and treated seeds suggests that different mechanisms modulate ABA content in response to different stresses, and that a specific ABA-signalling pathway regulates germination. Proteomic analysis revealed 46 proteins differentially expressed between treated and untreated seeds; 14 of these proteins were subsequently identified by mass spectrometry. Several of the proteins identified are important factors in the stress response, and are involved in such diverse functions as lipid metabolism, protein folding and chromatin protection. Lastly, an analysis of the phosphoproteome maps showed that the function of many proteins in seeds subjected to temperature treatment is modulated through post-translational modifications.