Transient species driving ecosystem multifunctionality: Insights from competitive interactions between rocky intertidal mussels.

Anthropogenic biodiversity loss poses a significant threat to ecosystem functioning worldwide. Numerically dominant and locally rare (i.e., transient) species are key components of biodiversity, but their contribution to multiple ecosystem functions (i.e., multifunctionality) has been seldomly assessed in marine ecosystems. To fill this gap, here we analyze the effects of a dominant and a transient species on ecosystem multifunctionality. In an observational study conducted along ca. 200 km of the southeastern Pacific coast, the purple mussel Perumytilus purpuratus numerically dominated the mid-intertidal and the dwarf mussel Semimytilus patagonicus exhibited low abundances but higher recruitment rates. In laboratory experiments, the relative abundances of both species were manipulated to simulate the replacement of P. purpuratus by S. patagonicus and five proxies for ecosystem functions-rates of clearance, oxygen consumption, total biodeposit, organic biodeposit, and excretion-were analyzed. This replacement had a positive, linear, and significant effect on the combined ecosystem functions, particularly oxygen consumption and excretion rates. Accordingly, S. patagonicus could well drive ecosystem functioning given favorable environmental conditions for its recovery from rarity. Our study highlights therefore the key role of transient species for ecosystem performance. Improving our understanding of these dynamics is crucial for effective ecosystem conservation, especially in the current scenario of biological extinctions and invasions.

Understanding the Symbiotic Relationship between the Sea Urchin Loxechinus albus (Molina, 1782) and the Pea Crab Pinnaxodes chilensis (H. Milne Edwards, 1837): a Potential Parasitism.

The echinoderm Loxechinus albus has a symbiotic relationship with the pinnotherid crustacean Pinnaxodes chilensis. Females of the crustacean develop in the terminal section of the sea urchin's digestive system, remaining there for life. This relationship has been suggested as commensalism. However, a potential negative impact on gonadal development and on the morphology of the sea urchin's digestive system suggest that it is instead parasitic. To study if there is a negative impact of the crustacean symbiont on the host, specimens of L. albus of all sizes were collected from a rocky shore in southern Chile. The gonadal and somatic tissues of sea urchins that were and were not harboring the pinnotherid were weighed and compared. Our results show that the presence of the pinnotherid was related to sea urchin gonads of lower biomass, decreased gonadosomatic index levels, and alterations in the morphology of the terminal portion of the host digestive system. The lower gonadal biomass suggests a negative impact on gamete production as well as a diversion of energy due to changes of the digestive system tissues and the potential consumption of algal food by the resident crustacean. These results suggest that the prolonged relationship between these two species is one of parasitism rather than one of commensalism.

Specific plasticity of the anemone Anthopleura hermaphroditica to intertidal and subtidal environmental conditions of the Quempillén estuary.

The cellular capacity of marine organisms to address rapid fluctuations in environmental conditions is decisive, especially when their bathymetric distribution encompasses intertidal and subtidal zones of estuarine systems. To understand how the bathymetric distribution determines the oxidative damage and antioxidant response of the estuarine anemone Anthopleura hermaphroditica, individuals were collected from upper intertidal and shallow subtidal zones of Quempillén River estuary (Chile), and their response analysed in a fully orthogonal, multifactorial laboratory experiment. The organisms were exposed to the effects of temperature (10°C and 30°C), salinity (10 ppt and 30 ppt) and radiation (PAR, > 400-700 nm; PAR+UV-A, > 320-700 nm; PAR+UV-A+UV-B, > 280-700 nm), and their levels of lipid peroxidation, protein carbonyl and total antioxidant capacity were determined. The results indicated that the intertidal individuals of A. hermaphroditica presented higher levels of tolerance to the stressful ranges of temperature, salinity, and radiation than individuals from the subtidal zone, which was evident from their lower levels of oxidative damage to lipids and proteins. These results were consistent with increased levels of total antioxidant capacity observed in subtidal organisms. Thus intertidal individuals could have greater plasticity to environmental variations than subtidal individuals. Future studies are needed to understand the mechanisms underlying stress adaptation in individuals from this estuarine anemone subjected to different environmental stressors during their life cycles.

The interactive effect of anti-sea lice pesticide azamethiphos and temperature on the physiological performance of the filter-feeding bivalve Ostrea chilensis: A non-target species.

The pesticide azamethiphos used by the salmon industry to treat sea lice, is applied as a bath and subsequently discharged into the sea. The effects of azamethiphos concentration (0, 15 and 100 µg L-1) on the physiology of the Chilean oyster (Ostrea chilensis) at two temperatures (12 and 15 °C) was examined. In all azamethiphos treatments, oysters kept at 15 °C had clearance rates (CR) higher than oysters kept at 12 °C. The oxygen consumption rate (OCR) increased at higher temperatures, except with 100 µg L-1 of azamethiphos, where no changes were observed. Sixty days after the exposure, survival rates of 91 and 79% (15 and 100 µg L-1, respectively), were observed compared to the controls, a situation independent of the experimental temperature. The interaction between temperature and pesticide has detrimental effects on the physiological performance and survival of O. chilensis, and these effects should also be assessed for other non-target species.

Effects of Ultraviolet Radiation on Sediment Burial Parameters and Photo-Oxidative Response of the Intertidal Anemone Anthopleura hermaphroditica.

Anthopleura hermaphroditica is an intertidal anemone that lives semi-buried in soft sediments of estuaries and releases its brooded embryos directly to the benthos, being exposed to potentially detrimental ultraviolet radiation (UVR) levels. In this study, we investigated how experimental radiation (PAR: photosynthetically active radiation; UVA: ultraviolet A radiation; and UVB: ultraviolet B radiation) influences burrowing (time, depth and speed) in adults and juveniles when they were exposed to PAR (P, 400-700 nm), PAR + UVA (PA, 315-700 nm) and PAR + UVA + UVB (PAB, 280-700 nm) experimental treatments. The role of sediment as a physical shield was also assessed by exposing anemones to these radiation treatments with and without sediment, after which lipid peroxidation, protein carbonyls and total antioxidant capacity were quantified. Our results indicate that PAB can induce a faster burial response compared to those anemones exposed only to P. PAB increased oxidative damage, especially in juveniles where oxidative damage levels were several times higher than in adults. Sediment offers protection to adults against P, PA and PAB, as significant differences in their total antioxidant capacity were observed compared to those anemones without sediment. Conversely, the presence or absence of sediment did not influence total antioxidant capacity in juveniles, which may reflect that those anemones have sufficient antioxidant defenses to minimize photooxidative damage due to their reduced tolerance to experimental radiation. Burrowing behavior is a key survival skill for juveniles after they have been released after brooding.

Non-consumptive effects of a predatory snail (Acanthina monodon) on a dominant habitat-forming mussel species (Perumytilus purpuratus).

Predators can influence prey through direct consumption as well as through non-consumptive effects (NCEs). NCEs usually occur mediated by behavioral changes in the prey upon detection of predator cues. Such changes may involve reduction of feeding with a variety of physiological consequences. We evaluated NCEs from an intertidal predatory snail (Acanthina monodon) on a dominant habitat-forming mussel species (Perumytilus purpuratus) from the southeastern Pacific coast. We tested whether A. monodon exerts negative NCEs on clearance rate, oxygen consumption rate, biodeposit production, and between-valve gap size in P. purpuratus. We found that waterborne predator cues triggered a decrease in these variables except biodeposit production. However, the organic content of the biodeposits increased in the presence of predator cues. The snail's physical contact with the mussels strengthened the negative NCEs on between-valve gap size. Since P. purpuratus is a dominant filter-feeder and foundation species in rocky intertidal habitats, predator NCEs on this species might indirectly influence ecosystem-level processes and community structure.

Community-wide consequences of nonconsumptive predator effects on a foundation species.

Predators can exert nonconsumptive effects (NCEs) on prey, which often take place through prey behavioural adjustments to minimise predation risk. As NCEs are widespread in nature, interest is growing to determine whether NCEs on a prey species can indirectly influence several other species simultaneously, thus leading to changes in community structure. In this study, we investigate whether a predator can exert NCEs on a foundation species and indirectly affect community structure. Through laboratory experiments, we first tested whether the predatory marine snail Acanthina monodon exerts negative NCEs on larviphagy (consumption of pelagic larvae) and phytoplankton filtration rates of the mussel Perumytilus purpuratus, an intertidal foundation species. These hypotheses stem from the notion that mussels may decrease feeding activities in the presence of predator cues to limit detection by predators. Afterwards, a field experiment tested whether the presence of A. monodon near mussel beds leads to higher colonisation rates of invertebrates that reproduce through pelagic larvae (expected under a lower larviphagy in P. purpuratus) and to a lower algal biomass on P. purpuratus shells (expected under a lower metabolite excretion in the mussels), thereby changing the community structure of the species typically found in P. purpuratus beds. The laboratory experiments revealed that waterborne cues from A. monodon limit the larviphagy and filtration rates of P. purpuratus. In turn, the field experiment showed that A. monodon cues led to greater abundances of barnacles and bivalves and a lower algal biomass in P. purpuratus beds, thus altering community structure. Overall, this study shows that a predator can indirectly affect community structure through NCEs on an invertebrate foundation species. As invertebrate foundation species are ubiquitous worldwide, understanding predator NCEs on these organisms could help to better understand community regulation in systems structured by such species.

Impact of short-term elevated temperature stress on winter-acclimated individuals of the marine gastropod Crepidula fornicata.

The intertidal zone is an especially stressful thermal habitat, typically exposing residents to air temperatures for up to 6 h at a time, twice daily. Tolerance to elevated temperatures has been particularly well-studied for a variety of intertidal species, especially with regard to upper thermal limits during summers. However, in recent years, as climates have been changing around the world, temperate zone intertidal organisms have sometimes been exposed to periods of unusually high air temperatures during the winter. The present study sought to examine the impact of elevated temperatures on survival and clearance rates of winter-acclimated intertidal individuals of the sedentary marine suspension-feeding gastropod Crepidula fornicata. Individuals were collected intertidally from Nahant, Massachusetts from late January to early April each year for 5 years, maintained in the laboratory at the acclimation temperature of 6 °C, and exposed in the laboratory for 3 h to temperatures as high as 37 °C in seawater either once or twice, 24 h apart. Although mean clearance rates were substantially reduced for at least the next 12-24 h after individuals were returned to the 6 °C control condition following exposures to elevated temperatures as low as 21-26 °C, we saw little mortality even following two 3 h exposures to 35 °C, or single exposures to 37 °C. Mortality was substantial, however, in one experiment following a double exposure to 37 °C. Smaller individuals (~5-12 mm in shell length) were somewhat more sensitive to the thermal stress than adults were. Intertidal members of C. fornicata in Massachusetts seem well-prepared to deal with the increasing range of winter air temperatures associated with the global climate confusion predicted for future years. Additional studies will be required to understand the physiological and biochemical mechanisms used by winter-acclimated individuals of this species to tolerate such periodic substantial temperature increases of 29-31 °C.

Plastic response of the oyster Ostrea chilensis to temperature and pCO2 within the present natural range of variability.

Estuaries are characterized by high fluctuation of their environmental conditions. Environmental parameters measured show that the seawater properties of the Quempillén estuary (i.e. temperature, salinity, pCO2, pH and ΩCaCO3) were highly fluctuating and related with season and tide. We test the effects of increasing temperature and pCO2 in the seawater on the physiological energetics of the bivalve Ostrea chilensis. Juvenile oysters were exposed to an orthogonal combination of three temperatures (10, 15, and 20°C) and two pCO2 levels (~400 and ~1000 µatm) for a period of 60 days to evaluate the temporal effect (i.e. 10, 20, 30, 60 days) on the physiological rates of the oysters. Results indicated a significant effect of temperature and time of exposure on the clearance rate, while pCO2 and the interaction between pCO2 and the other factors studied did not show significant effects. Significant effects of temperature and time of exposure were also observed on the absorption rate, but not the pCO2 nor its interaction with other factors studied. Oxygen consumption was significantly affected by pCO2, temperature and time. Scope for growth was only significantly affected by time; despite this, the highest values were observed for individuals subject to to 20°C and to ~1000 µatm pCO2. In this study, Ostrea chilensis showed high phenotypic plasticity to respond to the high levels of temperature and pCO2 experienced in its habitat as no negative physiological effects were observed. Thus, the highly variable conditions of this organism's environment could select for individuals that are more resistant to future scenarios of climate change, mainly to warming and acidification.

The filter-feeding bivalve Mytilus chilensis capture pelagic stages of Caligus rogercresseyi: A potential controller of the sea lice fish parasites.

The copepod Caligus rogercresseyi is an ectoparasite of several salmonid species. The pumping activity of filter-feeding molluscs could reduce the abundance of copepod dispersive larval stages in the water column. In this research, nauplius II and copepodid larvae of C. rogercresseyi were exposed to filtering mussels (Mytilus chilensis) of different sizes. These mussels were able to filter both larval stages, although they were more efficient in catching nauplius II. The fact that nauplius II were ingested more efficiently could be explained by their smaller size, lower swimming velocity (escape) and longer resting times between movements, when they were exposed to the influx of water around the inhalant area of the mussels. Larger mussels were more effective filtering C. rogercresseyi larvae due to their larger inhalant area and the related water influx. Additionally, the results suggest that larvae captured by the mussels can be incorporated into pseudofaeces or ingested and then released as part of the faeces. Thus, high concentrations of M. chilensis surrounding salmon farms may act as biological barriers, reducing the density of copepod dispersive larval stages and, thus, salmon infestations.

Reproductive biology of the encapsulating, brooding gastropod Crepipatella dilatata Lamarck (Gastropoda, Calyptraeidae).

Among calyptraeid gastropods, males become females as they get older, and egg capsules containing developing embryos are maintained beneath the mother's shell until the encapsulated embryos hatch. Crepipatella dilatata is an interesting biological model considering that is an estuarine species and thus periodically exposed to elevated environment-physiological pressures. Presently, there is not much information about the reproductive biology and brooding parameters of this gastropod. This paper describes field and laboratory observations monitoring sex changes, brooding frequencies, sizes of brooding females, egg mass characteristics, and embryonic hatching conditions. Our findings indicate that C. dilatata is a direct-developing protandric hermaphrodite, changing from male to female when individuals were between 18 and 20 mm in shell length. At our study site in Quempillén estuary, females were found to be brooding almost continuously throughout the year, having an average maximum of 85% of simultaneous brooding, with a short rest from April through June. No relationship was found between the number of capsules per egg mass and the size of the brooding female. However, capsule size and the number of embryos and nurse eggs were strongly related to female size. The offspring hatched with an average shell length > 1 mm. About 25% of the hatched capsules were found to contain both metamorphosed (juveniles) and non-metamorphosed (veliger) individuals. The sizes of the latter were < 1000 µm. The length of hatching juveniles was inversely related to the number of individuals per capsule, which seems related to differences in the availability of nurse eggs per embryo. Although fecundity per reproductive event of this species is relatively low (maximum approx. 800 offspring per egg mass) compared with those of calyptraeid species showing mixed development, the overall reproductive potential of C. dilatata seems to be high considering that females can reproduce up to 5 times per year, protecting their encapsulated embryos from physical stresses until well-developed juveniles are released into the population, avoiding a dangerous pelagic period prior to metamorphosis.

Trophic transfer of paralytic shellfish toxin (PST): Physiological and reproductive effects in the carnivorous gastropod Acanthina monodon (Pallas, 1774).

Harmful algal blooms can adversely affect different levels of the trophic chain, from primary consumers, such as bivalve molluscs, to higher links such as large fish, birds and mammals, including humans. Among secondary consumers, it has been described that carnivorous gastropods can accumulate these toxins when they prey on bivalves that have been exposed to toxic microalgae; these could also harm human health. In Chile, frequent events of harmful algal blooms caused by the dinoflagellate Alexandrium catenella have been described. This organism produces paralytic shellfish toxin (PST) which has been identified in some carnivorous gastropods. The objective of this research was to identify the physiological and reproductive response of the carnivorous gastropod Acanthina monodon fed on the Mytilid Perumytilus purpuratus, which had previously been maintained on a diet containing PST. Specimens of A. monodon showed a decrease in ingestion and absorption rate when they consumed PST indirectly through their diet. The oxygen consumption rate was also affected by the diet-time interaction. The variations of these parameters were reflected in the scope for growth, since the available energy was lower in gastropods exposed to toxic diet. Consumption of PST had a negative effect on the reproduction of A. monodon, since intoxicated adults presented lower egg-masses and delayed start of oviposition. We observed a delay in the development of the embryos inside the capsules, and a lower number of hatched juveniles, although these few juveniles from intoxicated parents accomplished higher growth rates during the next 6 months. We may therefore suggest that toxin transfer, from harmful microalgae through the trophic chain, can generate deleterious effects on the physiological energetics of the organisms that consume them, affecting their reproductive capacity and early ontogenetic development.

Ocean acidification exacerbates the effects of paralytic shellfish toxins on the fitness of the edible mussel Mytilus chilensis.

High latitudes are considered particularly vulnerable to ocean acidification, since they are naturally low in carbonate ions. The edible mussel Mytilus chilensis is a common calcifier inhabiting marine ecosystems of the southern Chile, where culturing of this species is concentrated and where algal blooms produced by the toxic dinoflagellate A. catenella are becoming more frequent. Juvenile Mytilus chilensis were exposed to experimental conditions simulating two environmental phenomena: pCO2 increase and the presence of paralytic shellfish toxins (PST) produced by the dinoflagellate Alexandrium catenella. Individuals were exposed to two levels of pCO2: 380 µatm (control condition) and 1000 µatm (future conditions) over a period of 39 days (acclimation), followed by another period of 40 days exposure to a combination of pCO2 and PST. Both factors significantly affected most of the physiological variables measured (feeding, metabolism and scope for growth). However, these effects greatly varied over time, which can be explained by the high individual variability described for mussels exposed to different environmental conditions. Absorption efficiency was not affected by the independent effect of the toxic diet; however, the diet and pCO2 interaction affected it significantly. The inhibition of the physiological processes related with energy acquisition by diets containing PST, may negatively impact mussel fitness, which could have important consequences for both wild and cultured mussel populations, and thus, for socioeconomic development in southern Chile.

Pre-ingestive selection capacity and endoscopic analysis in the sympatric bivalves Mulinia edulis and Mytilus chilensis exposed to diets containing toxic and non-toxic dinoflagellates.

This study investigates the effects of toxic and non-toxic dinoflatellates on two sympatric bivalves, the clam Mulinia edulis and the mussel Mytilus chilensis. Groups of bivalves were fed one of three diets: (i) the toxic paralytic shellfish producing (PSP) Alexandrium catenella + Isochrysis galbana; (ii) the non-toxic Alexandrium affine + Isochrysis galbana and (iii) the control diet of Isochrysis galbana. Several physiological traits were measured, such as, clearance rate, pre-ingestive selection efficiency and particle transport velocity in the gill. The clearance rates of both M. chilensis and M. edulis showed a significant reduction when fed a mixed toxic diet of 50% Alexandrium catenella: 50% Isochrysis galbana. Similarly, when both species of bivalves were fed with the non-toxic diet (50% A. affine: 50% I. galbana), clearance rate was significantly lower compared with a diet of 100% I. galbana. Under all the experimental diets, M. chilensis showed higher clearance rate values, slightly more than double that of M. edulis. M. edulis and M. chilensis have the ability to select particles at the pre-ingestive level, thus eliminating a larger proportion of the toxic dinoflagellate A. catenella as well as the non-toxic A. affine in the form of pseudofaeces. Higher values of selection efficiency were registered in M. edulis than in M. chilensis when exposed to the toxic diet. Similar results were observed when these two species were exposed to the diet containing the non-toxic dinoflagellate, explained by the fact that the infaunal Mulinia edulis is adapted to dealing with larger particle sizes and higher particle densities (Navarro et al., 1993). The lower transport particle velocity observed in the present work for both species, is related to the reduced clearance rate, the higher particle concentration, and the presence of larger, toxic dinoflagellates. In addition, the species differ in their feeding responses to diets, with and without A. catenella or A. affine, largely reflecting their adaptations to different environmental conditions. The results suggest that the presence of a dinoflagellate bloom, whether toxic or non-toxic spp in Yaldad Bay, is likely to have a greater impact on the Mytilus chilensis than the infaunal Mulinia edulis, based on the combined effects on clearance rate, selection efficiency and particle transport velocity.

Growing Safe: Acute Size Escape from Desiccation in Juvenile Crepipatella peruviana (Mollusca: Gastropoda).

Desiccation is an important limiting factor in the intertidal zone. Generally decreasing seaward, desiccation stress can also be alleviated in wet microhabitats. Juvenile snails are generally more susceptible to desiccation than adults, and, for some species, juveniles must therefore hide in microhabitats to survive emersion. The transition from hiding in safe microhabitats to being able to survive fully exposed for the duration of low tide is not well documented. In this study, we investigated the influence of size on desiccation tolerance in juveniles of the calyptraeid gastropod Crepipatella peruviana to determine the size at which they can first survive exposure to air. Juveniles 2-13 mm long were exposed to 75% or 100% relative humidity for 0.5-6.5 hours. Juveniles smaller than 5 mm in shell length did not survive emersion at 75% relative humidity for even 0.5 hours; surprisingly, most also perished after short exposures to air at 100% relative humidity, suggesting that something other than desiccation stress may also be at play. In marked contrast, 82% of juveniles larger than 6 mm in shell length survived exposure to 75% relative humidity for the full 6.5 hours. In a field survey, no juveniles smaller than 9 mm were found on exposed rock but rather were found only in wet microhabitats. We suggest that the clearly defined size escape from desiccation may reflect a change in gill functioning or a newfound ability to retain water more effectively within the mantle cavity at low tide.

Effects of Embryonic Exposure to Salinity Stress or Hypoxia on Post-metamorphic Growth and Survival of the Polychaete Capitella teleta.

Although a good number of studies have investigated the impact of larval experience on aspects of post-metamorphic performance, only a few have considered the potential impact of stresses experienced by brooded embryos. In this study we separately investigated the impact of salinity stress (as low as 10) and hypoxia (1 ml O2 l-1) experienced by brooded embryos of the deposit-feeding polychaete Capitella teleta on hatching success, metamorphosis, post-metamorphic survival, and post-metamorphic growth. Salinity reduction from 30 to 10 or 15 reduced relative hatching success, presumably by reducing embryonic survival, but generally had no negative latent effects on juvenile survival or growth. Prolonged exposure to hypoxic conditions had no negative effects, as seen on measurements recorded, other than abandonment of brood tubes by some females. There were no negative effects on days to emergence from brood tubes, numbers of larvae emerging from brood tubes, juvenile survival, or juvenile growth. Future studies should consider the potential role of maternal behavior in protecting embryos from at least short-term exposures to hypoxia, and the capacity for anaerobic metabolism in both embryos and adults of this species.

Role of the Substrate in Feeding and Growth of the Marine Suspension-Feeding Gastropods Crepidula fornicata and Crepipatella peruviana.

Unlike lamellibranch bivalves, suspension-feeding calyptraeid gastropods lack siphons and paired shell valves to regulate water inflow. This study was designed to determine if calyptraeid gastropods use the solid surface to which they attach to facilitate food particle capture. Juveniles of both Crepidula fornicata and Crepipatella peruviana were maintained with phytoplankton for 3 to 6 wk in the laboratory, either attached to solid substrate or without solid substrate. Individuals of C. fornicata and C. peruviana that were reared on solid substrate grew about five to ten times more, or two times more, respectively, than those deprived of solid substrate. Final tissue weights were also significantly greater for individuals of both species that had been reared on solid substrate. For the two species, phytoplankton clearance rates were about two to three times higher for individuals attached to solid substrate than for those without solid substrate; rates of food cord production from the gills were also significantly higher. About 50% of C. peruviana that were deprived of solid substrate died during the first 3 wk of observation, and about 60% were dead by 6 wk. In contrast, most individuals of C. peruviana that were attached to solid substrate survived for the entire 6-wk study period, and all of the C. fornicata survived whether or not they were attached to solid substrate. The solid substrate to which calyptraeid gastropods attach clearly plays an important role in their feeding biology, although the precise role remains to be explored.

Brooding strategy in fluctuating salinity environments: oxygen availability in the pallial cavity and metabolic stress in females and offspring in the Chilean oyster Ostrea chilensis.

Organisms that encounter stressful situations in nature often cope using behavioral (e.g., avoidance) or physiological tactics. In sessile mollusks, the only available behavioral option in dealing with salinity stress is to "clam up", isolating their tissues from the environment. Though effective in the short term, prolonged isolation can have detrimental physiological consequences, particularly for females brooding embryos in a mantle cavity that is isolated from the external environment. In the Quempillén estuary, the Chilean oyster, Ostrea chilensis, spent nearly one-third of its brooding season at salinities low enough to cause female isolation. When females thus isolated themselves, the dissolved oxygen in their mantle cavity fluid dropped to hypoxic levels within 10 min. In females that were brooding embryos, this depletion of oxygen was not uniform: oxygen was depleted more quickly in the palp region (where embryos accumulate) than in the inhalant region. Additionally, oxygen was reduced even more quickly in the palp region when females were brooding late-stage embryos, which consumed oxygen significantly more quickly than embryos in earlier developmental stages. Finally, O. chilensis used anaerobic metabolism to cope with the hypoxia induced by isolation, as lactate accumulated in the tissues of both females (brooding > non-brooding) and embryos (late stage > early stage). Our findings demonstrate the trade-off between an adaptive avoidance behavior (clamming up) and the potentially detrimental consequences brought on by such a behavior (hypoxia). Cycling of embryos throughout the mantle cavity by deliberate female pumping keeps them from accumulating in the area between the palps, forestalling the creation of hypoxic conditions there. In addition, the capacity for anaerobic metabolism by both females and their embryos should help them tolerate the low oxygen levels that do eventually arise when the pallial cavity is isolated from the surrounding environment during long periods of reduced ambient salinity.

Brooding in the Chilean oyster Ostrea chilensis: unexpected complexity in the movements of brooded offspring within the mantle cavity.

Brooding in invertebrates serves to protect embryos from stressful external conditions by retaining progeny inside the female body, effectively reducing the risk of pelagic stages being exposed to predation or other environmental stressors, but with accompanying changes in pallial fluid characteristics, including reduced oxygen availability. Brooded embryos are usually immobile and often encapsulated, but in some Ostrea species the embryos move freely inside the female pallial cavity in close association with the mother's gills for as long as eight weeks. We used endoscopic techniques to characterize the circulation pattern of embryos brooded by females of the oyster, Ostrea chilensis. Progeny at embryonic and veliger stages typically circulated in established patterns that included the use of dorsal and ventral food grooves (DFG, VFG) to move anteriorly on the gills. Both embryos and veligers accumulated around the mother's palps, and remained there until an active maternal countercurrent moved them to the gill inhalant area. Both food grooves were able to move embryos, veligers, and food-particle aggregates anteriorly, but the DFG was more important in progeny transport; early embryos were moved more rapidly than veligers in the DFG. A microcirculation pattern of embryos was apparent when they were moved by gill lamellae: when they were close to the VFG, most embryos lost gill contact and "fell" down to the DFG. Those that actually reached the DFG moved anteriorly, but others came into contact with the base of the lamellae and again moved towards the VFG. The circulation pattern of the progeny appears well-suited for both cleaning them and directing them posteriorly to an area where there is more oxygen and food than in the palp region. This process for actively circulating progeny involves the feeding structures (gill and palps) and appears to be energetically costly for the female. It also interferes with feeding, which could explain the poor energy balance previously documented for brooding females of this species.

Effects of low salinity on adult behavior and larval performance in the intertidal gastropod Crepipatella peruviana (Calyptraeidae).

Shallow-water coastal areas suffer frequent reductions in salinity due to heavy rains, potentially stressing the organisms found there, particularly the early stages of development (including pelagic larvae). Individual adults and newly hatched larvae of the gastropod Crepipatella peruviana were exposed to different levels of salinity stress (32(control), 25, 20 or 15), to quantify the immediate effects of exposure to low salinities on adult and larval behavior and on the physiological performance of the larvae. For adults we recorded the threshold salinity that initiates brood chamber isolation. For larvae, we measured the impact of reduced salinity on velar surface area, velum activity, swimming velocity, clearance rate (CR), oxygen consumption (OCR), and mortality (LC50); we also documented the impact of salinity discontinuities on the vertical distribution of veliger larvae in the water column. The results indicate that adults will completely isolate themselves from the external environment by clamping firmly against the substrate at salinities ≤24. Moreover, the newly hatched larvae showed increased mortality at lower salinities, while survivors showed decreased velum activity, decreased exposed velum surface area, and decreased mean swimming velocity. The clearance rates and oxygen consumption rates of stressed larvae were significantly lower than those of control individuals. Finally, salinity discontinuities affected the vertical distribution of larvae in the water column. Although adults can protect their embryos from low salinity stress until hatching, salinities <24 clearly affect survival, physiology and behavior in early larval life, which will substantially affect the fitness of the species under declining ambient salinities.