Upwelling as a stressor event during embryonic development: Consequences for encapsulated and early juvenile stages of the marine gastropod Acanthina monodon.

Upwelling phenomena alter the physical and chemical parameters of the sea's subsurface waters, producing low levels of temperature, pH and dissolved oxygen, which can seriously impact the early developmental stages of marine organisms. To understand how upwelling can affect the encapsulated development of the gastropod Acanthina monodon, capsules containing embryos at different stages of development (initial, intermediate and advanced) were exposed to upwelling conditions (pH = 7.6; O2 = 3 mg L-1; T° = 9 °C) for a period of 7 days. Effects of treatment were determined by estimating parameters such as time to hatching, number of hatchlings per capsule, percentage of individuals with incomplete development, and shell parameters such as shell shape and size, shell strength, and the percentage of the organic/inorganic content. We found no significant impacts on hatching time, number of hatchlings per capsule, or percentage of incomplete development in either the presence or absence of upwelling, regardless of developmental stage. On the other hand, latent effects on encapsulated stages of A. monodon were detected in embryos that had been exposed to upwelling stress in the initial embryonic stage. The juveniles from this treatment hatched at smaller sizes and with higher organic content in their shells, resulting in a higher resistance to cracking 30 days after hatching, due to greater elasticity. Geometric morphometric analysis showed that exposure to upwelling condition induced a change in the morphology of shell growth in all post-hatching juveniles (0-30 days), regardless of embryonic developmental stage at the time of exposure. Thus, more elongated shells (siphonal canal and posterior region) and more globular shells were observed in newly hatched juveniles that had been exposed to the upwelling condition. The neutral or even positive upwelling exposure results suggests that exposure to upwelling events during the encapsulated embryonic phase of A. monodon development might not have major impacts on the future juvenile stages. However, this should be taken with caution in consideration of the increased frequency and intensity of upwelling events predicted for the coming decades.

Embryonic encapsulated development of the gastropod Acanthina monodon is impacted by future environmental changes of temperature and pCO2.

Egg capsules of the gastropod Acanthina monodon were maintained during the entire period of encapsulated development at three temperatures (10, 15, 20 °C) and two pCO2 levels (400, 1200 µatm). Embryos per capsule, size at hatching, time to hatching, embryonic metabolic rates, and the resistance of juveniles to shell breakage were quantified. No embryos maintained at 20 °C developed to hatching. The combination of temperature and pCO2 levels had synergistic effects on hatching time and developmental success, antagonistic effects on number of hatchlings per capsule, resistance to juvenile shell cracking and metabolism, and additive effect on hatching size. Juveniles hatched significantly sooner at 15 °C, independent of the pCO2 level that they had been exposed to, while individuals hatched at significantly smaller sizes if they had been held under 15 °C/1200 µatm rather than at 10 °C/low pCO2. Embryos held at the higher pCO2 had a significantly greater percentage of abnormalities. For capsules maintained at low pCO2 and 15 °C, emerging juveniles had less resistance to shell breakage. Embryonic metabolism was significantly higher at 15 °C than at 10 °C, independent of pCO2 level. The lower metabolism occurred in embryos maintained at the higher pCO2 level. Thus, in this study, temperature was the factor that had the greatest effect on the encapsulated development of A. monodon, increasing the metabolism of the embryos and consequently accelerating development, which was expressed in a shorter intracapsular development time, but with smaller individuals at hatching and a lower resistance of their shells to breakage. On the other hand, the high pCO2 level suppressed metabolism, prolonged intracapsular development, and promoted more incomplete development of the embryos. However, the combination of the two factors can mitigate--to some extent--the adverse effects of both incomplete development and lower resistance to shell breakage.

Energetic trade-offs: Implications for selection between two bivalve prey species by a carnivorous muricid gastropod.

Active predators obtain energy and nutrients from prey through complex processes in which the energy gained must exceed the energy invested in finding and ingesting the prey. In addition, the amount of energy available will vary with the prey that are selected for consumption. The muricid gastropod Acanthina monodon inhabits rocky shores, where it routinely feeds on the mytilids Semimytilus algosus and Perumytilus purpuratus. In this study, S. algosus was highly preferred by the predator (over 90% were eaten) versus P. purpuratus (only 9% were eaten) when offered a mixed diet. The energetic cost of attacking one S. algosus individual was 91 J bivalve-1 while for P. purpuratus it was slightly higher: 95 J bivalve-1. Also, whereas A. monodon required on average 19 h to consume S. algosus, successful attacks on P. purpuratus required about 32% more time (25 h). In addition, a longer resting time was needed by the predator after preying on P. purpuratus before it initiated another attack. Moreover, the active metabolic costs associated with successfully attacking the prey increased 3.2 times over the basal metabolic costs when attacking S. algosus, but only by 2.5 times when attacking P. purpuratus. The calculations associated with preying on each species showed that the energetic gain per unit time likely accounts for the predator's preference for attacking S. algosus, even though predation on both species provided net energy gains for the predator. However, as S. algosus occurs seasonally at our study site, P. purpuratus would probably also be consumed due to its constant availability throughout the whole year.

Relationship between over-crowding within egg capsules of the marine gastropod Acanthina monodon and prospects for juvenile success.

Encapsulated development with extraembryonic yolk may lead to competition for nutrients within egg capsules. In this research, different degrees of competition among embryos in subtidal egg capsules of Acanthina monodon resulted in considerable differences in hatching size. For newly hatched juveniles, individuals hatching from less crowded egg capsules showed better survival, larger SL, higher rates of oxygen consumption, and higher rates of food consumption. However, by 28 days after hatching, the largest surviving juveniles were the best-performing individuals, regardless of the initial embryo density within the capsules. In summary, more crowded egg capsules resulted in poorer survival. These findings may help to explain the variability seen in juvenile success in some field populations; much of that variation may reflect stressful experiences that the new recruits have had during the early stages of their encapsulated development.

Respiratory and desiccation constraints during encapsulated intertidal development of the marine gastropod Acanthina monodon.

Acanthina monodon commonly deposits its egg capsules in the intertidal zone. Capsule aerial exposure during low-tide can impact oxygen consumption rates (OCR) of embryos and intracapsular oxygen availability, and expose embryos to desiccation. OCR increased as embryonic development progressed, and was greater when capsules were submerged in seawater than when exposed to air. Oxygen available within the capsule was always less than that available in the immediate external environment, whether capsules were immersed or exposed. The highest internal oxygen concentrations were recorded during periods of air exposure for embryos in more advanced development stages. When exposed to air, capsules lost water the fastest when they contained early embryos, and suffered the highest mortalities following exposure. Collectively, these data suggest that, although encapsulation helps the embryos to develop across wildly fluctuating environmental conditions, the amount of stress the embryos experience will vary depending on their exact positioning within the intertidal zone.

Growth Stasis and Limited Shell Calcification in Larvae of Cymatium parthenopeum During Trans-Atlantic Transport.

Larvae of the shallow-water marine gastropod Cymatium parthenopeum show no appreciable shell calcification and no demonstrable growth as they disperse across the Atlantic Ocean. Evidence of what appears to be physiological specialization for prolonged delay of metamorphosis was found in larvae of this prosobranch gastropod.

Legacy of Multiple Stressors: Responses of Gastropod Larvae and Juveniles to Ocean Acidification and Nutrition.

Ocean acidification poses a significant threat to calcifying invertebrates by negatively influencing shell deposition and growth. An organism's performance under ocean acidification is not determined by the susceptibility of one single life-history stage, nor is it solely controlled by the direct physical consequences of ocean acidification. Shell development by one life-history stage is sometimes a function of the pH or pCO2 levels experienced during earlier developmental stages. Furthermore, environmental factors such as access to nutrition can buffer organismal responses of calcifying invertebrates to ocean acidification, or they can function as a co-occurring stressor when access is low. We reared larvae and juveniles of the planktotrophic marine gastropod Crepidula fornicata through combined treatments of nutritional stress and low pH, and we monitored how multiple stressors endured during the larval stage affected juvenile performance. Shell growth responded non-linearly to decreasing pH, significantly declining between pH 7.6 and pH 7.5 in larvae and juveniles. Larval rearing at pH 7.5 reduced juvenile growth as a carryover effect. Larval rearing at pH 7.6 reduced subsequent juvenile growth despite the absence of a negative impact on larval growth, demonstrating a latent effect. Low larval pH magnified the impact of larval nutritional stress on competence for metamorphosis and increased carryover effects of larval nutrition on juvenile growth. Trans-life-cycle effects of larval nutrition were thus modulated by larval exposure to ocean acidification.

Volcanic ash in the water column: Physiological impact on the suspension-feeding bivalve Mytilus chilensis.

Ashes settling into the sea from volcanic explosions expose suspension-feeding species to reduced seston quality. Adults and juveniles of the mussel Mytilus chilensis were exposed for 15days to the phytoplankton Isochrysis galbana together with various concentrations of ashes. We then quantified impact on survival and physiology. Although no individuals died during the experiment, by the end of the study clearance rates and oxygen consumption rates had decreased substantially, and tissue weight of mussels exposed to the highest ash concentrations declined substantially. Gills showed no physical damage, but did show abundant mucus secretion in response to ash particles. Moreover, as the relative proportions of microalgae to ash in the diet decreased, individuals showed increasing preferential ingestion of microalgal particles. Increased ash content in the diet altered physiological rates and activated distinct particle selection with a high production of pseudofeces and high energy costs, with potential long-term consequences.

Possible Mechanisms of Hatching from Egg Capsules in the Gastropods Crepipatella dilatata and Crepipatella peruviana, Species with Different Modes of Early Development.

Many invertebrates enclose their embryos within egg capsules, from which the offspring hatch. In marine gastropods that brood their egg capsules, hatching could involve radular activity by the mother or by unhatched stages, increased osmotic concentration of the intracapsular fluid, or production of hatching enzymes. The present research sought to determine whether mechanical action by the brooding female or by the encapsulated embryos was involved in the hatching for two sympatric and closely related species of calyptraeid: Crepipatella dilatata, which exhibits direct development without free-living larvae, and Crepipatella peruviana, which releases free-living veliger larvae. We also considered the role that enzymatic action or osmotic changes in the intracapsular fluid might play in hatching. Using scanning electron micrograph analyses, we found no evidence that the well-developed, pre-hatching juvenile radula of C. dilatata played any role in the hatching process and that the radula of C. peruviana did not even develop until long after hatching; so there was no evidence of radular activity involved in the hatching of either species. For C. peruviana, the intracapsular fluid osmolality was always higher than that of the surrounding seawater, suggesting that there is a strong natural water inflow during development. Moreover, when egg capsules of C. peruviana were exposed to lower ambient salinities, the substantial entry of water correlated well with high percentages of hatching, particularly for egg capsules containing advanced veligers, suggesting that an osmotic mechanism may be involved in the hatching process of this species. In contrast, hatching in C. dilatata appeared to be enzymatically mediated.

Crepidula fornicata is not a first intermediate host for trematodes: who is?

Trematode larvae must generally invade a molluscan intermediate host, usually a gastropod, before they can reach reproductive maturity in another definitive host. The research literature to date has focused almost exclusively on the documented specificity between particular trematode species and particular molluscan hosts; little attention has been paid to gastropod species that do not appear to serve as hosts. We sampled Rhode Island and Massachusetts populations of the marine gastropod Crepidula fornicata to determine whether this widespread species serves as a first intermediate host for trematodes. We also sampled from the same habitat populations of Littorina littorea and Ilyanassa obsoleta, gastropods known to serve as first intermediate hosts for several trematode species. All individuals were examined by dissection for the presence of sporocysts, rediae, or developing cercariae. Although 4-28% of L. littorea (N=112) and I. obsoleta (N=84) were infected by larvae of at least one trematode species, no individuals of C. fornicata sampled from the same locations were so infected (N=136). A survey of the Biological Abstracts computer database indicates that snails in only about 10% of marine gastropod families are known to serve as first intermediate hosts for trematodes. We suggest that more attention be paid to marine gastropods that appear not to be infected by trematode miracidia. Such species may productively serve as new models for understanding trematode host specificity and gastropod resistance to infection.

How Do Temperature and Salinity Affect Relative Rates of Growth, Morphological Differentiation, and Time to Metamorphic Competence in Larvae of the Marine Gastropod Crepidula plana?

The influence of environmental conditions on rates of larval growth has been documented many times for various marine mollusks. But the factors that influence rates of morphological and physiological differentiation, particularly the rate at which larvae within a population become competent to metamorphose, remain obscure. In four experiments, we reared larvae of the gastropod Crepidula plana at 29°C, 25°C, and 20°C at 30 ppt salinity, and in two other experiments, in salinities between 4-30 ppt at 25°C. Rates of shell growth and morphological differentiation, and rates of becoming competent within populations were recorded. Larvae were considered to be competent to metamorphose if they could be stimulated to metamorphose by exposure to a high concentration of KCl (20 mM above ambient). Larvae consistently became competent faster at higher temperatures, but in only one of four experiments did temperature also consistently increase the rates of growth and morphological differentiation. Larvae took longer to become competent when reared at lower salinities, but the effects were poorly predicted by the influence of salinity on rates of growth and morphological differentiation. Competent larvae could also not be recognized by shell length; many individuals were competent at shell lengths of 600-800 µm, while many other individuals were still not competent at sizes exceeding 1000 µm. At 29°C, many individuals became competent at smaller sizes than those reared at lower temperatures. Presence of gill filaments or shell brims also did not correlate with individual metamorphic competence. The data suggest that growth rate, rate of morphological differentiation, and time required for larvae of C. plana to become competent can be uncoupled markedly by shifts in rearing conditions.

Histochemical glycogen and neutral lipid in Echinostoma trivolvis cercariae and effects of exogenous glucose on cercarial longevity.

Histochemical glycogen and neutral lipid studies were conducted on Echinostoma trivolvis cercariae maintained in artificial spring water (ASW) at 24-25 degrees C for up to 24 h after emergence from host snails. Treatment of whole cercariae by the periodic acid Schiff (PAS) reagent with or without 1% malt diastase showed that cercariae depleted glycogen mainly from the tail by 6 to 24 h postemergence. The posterior tip of the tail remained PAS positive and diastase fast suggesting the presence of mucopolysaccharides there. Fresh cercariae or those stained up to 24 h postemergence with Oil Red O showed the presence of neutral lipid droplets in the excretory system. There was no discernible difference in the size, abundance, or distribution of these droplets in fresh or aged cercariae. Cercariae maintained in ASW plus 1% glucose for 12 or 23 h showed no evidence of resynthesizing glycogen. Nevertheless, cercariae survived longer in 1% glucose than in either 0.0, 0.1 or 0.5% glucose; but only at 23 h were any differences statistically greater (one way ANOVA, P < 0.05).