The Interactive Effects of the Anti-Sea Lice Pesticide Azamethiphos and Temperature on Oxidative Damage and Antioxidant Responses in the Oyster Ostrea chilensis.

Azamethiphos is used in the salmon industry to treat sea lice and is subsequently discharged into the sea, which may affect non-target species (NTS). A rise in seawater temperature could enhance the sensitivity of NTS. Thus, in the present investigation, the combined effects of azamethiphos (0 µg L-1, 15 µg L-1 and 100 µg L-1) and temperature (12 °C and 15 °C) was assessed over time (7 days) in the gonads and gills of the oyster Ostrea chilensis, assessing its oxidative damage (lipid peroxidation and protein carbonyls) and total antioxidant capacity. Our results indicated that in gonads and gills, lipid peroxidation levels increased over time during exposure to both pesticide concentrations. Protein carbonyl levels in gills increased significantly in all experimental treatments; however, in gonads, only pesticide concentration and exposure time effected a significant increase in protein damage. In both, gill and gonad temperature did not influence oxidative damage levels. Total antioxidant capacity in gonads was influenced only by temperature treatment, whereas in the gills, neither temperature nor azamethiphos concentration influenced defensive responses. In conclusion, our results indicated the time of pesticide exposure (both concentrations) had a greater influence than temperature on the cellular damage in this oyster.

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.

Early development and metabolic rate of the sea louse Caligus rogercresseyi under different scenarios of temperature and pCO2.

Anthropogenic CO2 emissions have led to ocean acidification and a rise in the temperature. The present study evaluates the effects of temperature (10, 15 and 20 °C) and pCO2 (400 and 1200 µatm) on the early development and oxygen consumption rate (OCR) of the sea louse Caligus rogercresseyi. Only temperature has an effect on the hatching and development times of nauplius I. But both factors affected the development time of nauplius II (

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.

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.