Phenotypic plasticity is not a cline: Thermal physiology of an intertidal barnacle over 20° of latitude.

Our understanding of the plastic and evolutionary potential of ectothermic organisms and their populational impacts in the face of rapid global change remains limited. Studies attempting on the relationship between the magnitude of thermal variability across latitude and the degree of phenotypic plasticity exhibited by marine ectotherms are inconclusive. We state that the latter arises from the narrow range of thermal variability captured by the limited span of the latitudinal gradients studied to date. Using a mechanistic ecophysiological approach and a satellite-based assessment of the relevant environmental variables (i.e. temperature and food availability), we studied individuals of the intertidal barnacle Jehlius cirratus from seven local populations widely spread along the Humboldt current system that spanning two biogeographic regions. At the same time, we synthesized published information on the local abundance of our study species across a total of 76 sites representing 20° of latitude, and spanning from 18 to 42°S. We examined the effects of latitude and environmental variability on metabolic rate plasticity, thermal tolerance (thermal breadth and thermal safety margins) and their impacts on the abundance of this widespread marine invertebrate. We demonstrate that the phenotypic plasticity of metabolic rate in J. cirratus populations is not related to latitude. In turn, thermal breadth is explained by the temperature variability each population experiences. Furthermore, we found clinal variation with a poleward decrease of the critical thermal minimum, suggesting that episodic extreme low temperatures represent a ubiquitous selective force on the lower thermal limit for ectotherms. Across our study gradient, plasticity patterns indicate that populations at the equatorial extreme are more vulnerable to a warming climate, while populations located in the biogeographic transitional zone (i.e. high environmental heterogeneity), on the centre of the gradient, display higher levels of phenotypic plasticity and may represent a genetic buffer for the effects of ocean warming. Together, our results suggest the existence of a fitness trade-off involving the metabolic cost of plasticity and population density that is evident only across the vast latitudinal gradient examined.

Nuestro conocimiento del potencial plástico y evolutivo de organismos ectotérmicos y de los posibles impactos poblacionales a la luz del rápido cambio global sigue siendo limitado. Los estudios que relacionan la magnitud de la variabilidad térmica y el grado de plasticidad fenotípica a través de la latitud realizados en organismos ectotérmicos marinos no son concluyentes. Lo anterior creemos que es consecuencia del estrecho rango latitudinal y por consecuencia el menor rango de variabilidad térmica abarcado por los estudios previos. Utilizando un enfoque ecofisiológico mecanicista e información satelital de las variables ambientales relevantes (i.e., temperatura y disponibilidad de alimento), estudiamos individuos del cirripedio intermareal Jehlius cirratus a lo largo de siete poblaciones locales que se distribuyen ampliamente a lo largo del Sistema de la corriente de Humboldt abarcando dos regiones biogeográficas. Al mismo tiempo, sintetizamos la información publicada sobre la abundancia local de nuestro modelo de estudio en un total de 76 sitios que representan 20 grados de latitud y abarcan desde los 18° a los 42°S. Examinamos los efectos de la latitud y la variabilidad ambiental en la plasticidad de la tasa metabólica, la tolerancia térmica (i.e. amplitud térmica y márgenes de seguridad térmica) y los impactos en la abundancia de este invertebrado marino con amplia distribución geográfica. Demostramos que la plasticidad fenotípica de la tasa metabólica en poblaciones de J. cirratus no está relacionada con la latitud. A su vez, la amplitud térmica se explica por la variabilidad térmica que cada población experimenta. Además, encontramos un patrón de variación clinal con una disminución hacia los polos del crítico térmico mínimo, lo que sugiere que las temperaturas episódicas extremadamente bajas representan una fuerza selectiva ubicua en el límite térmico inferior para los ectotermos. A lo largo de nuestro gradiente estudiado, los patrones de plasticidad indican que las poblaciones en el extremo ecuatorial son más vulnerables al calentamiento, mientras que las poblaciones ubicadas en la zona de transición biogeográfica (i.e., alta heterogeneidad ambiental), en el centro del gradiente, muestran mayores niveles de plasticidad fenotípica, lo que puede representar un reservorio genético para los efectos del calentamiento de los océanos. Nuestros resultados sugieren la existencia de un compromiso en la adecuación biológica que involucra el costo metabólico de la plasticidad y la densidad de población que es sólo evidente dado el vasto gradiente latitudinal examinado.

Thermal physiological traits and plasticity of metabolism are sensitive to biogeographic breaks in a rock-pool marine shrimp.

Populations of broadly distributed species commonly exhibit latitudinal variation in thermal tolerance and physiological plasticity. This variation can be interrupted when biogeographic breaks occur across the range of a species, which are known to affect patterns of community structure, abundance and recruitment dynamics. Coastal biogeographic breaks often impose abrupt changes in environmental characteristics driven by oceanographic processes and can affect the physiological responses of populations inhabiting these areas. Here, we examined thermal limits, performances for heart rate and plasticity in metabolic rate of the intertidal shrimp Betaeus emarginatus from seven populations along its latitudinal range (∼3000 km). The distribution of this species encompass two breaks along the southeastern Pacific coast of Chile: the northern break is characterized by sharp discontinuities in upwelling regimes, and the southern break constitutes a major discontinuity in water conditions (temperature, pH, dissolved oxygen and nutrients), coastline topography and divergence of main oceanographic currents. For B. emarginatus, we found higher plasticity in metabolism at the sites sampled at the biogeographic breaks, and at the site subjected to seasonal upwelling. The variation in metabolic rate was not consistent with increasing latitude and it was not affected by breaks. The lower and upper thermal limits were lower in populations around breaks, although the optimum temperature decreased towards higher latitudes. Overall, whereas thermal limits and plasticity of metabolism are related to biogeographic breaks, metabolic rate is not related to increasing latitude or the presence of breaks in the sampled range.

Exploring physiological plasticity and local thermal adaptation in an intertidal crab along a latitudinal cline.

Intertidal organisms have evolved physiological mechanisms that enable them to maintain performance and survive during periods of severe environmental stress with temperatures close to their tolerance limits. The level of these adaptive responses in thermal physiology can vary among populations of broadly distributed species depending on their particular environmental context and genetic backgrounds. Here we examined thermal performances and reaction norms for metabolic rate (MR) and heart rate (HR) of seven populations of the porcelanid crab Petrolisthes violaceus from markedly different thermal environments across the latitudinal gradient of ~3000km. Physiological responses of this intertidal crab under common-garden conditions suggest the absence of local thermal adaptation along the geographic gradient (i.e., lack of latitudinal compensation). Moreover, thermal physiological sensitivities and performances in response to increased temperatures evidenced the existence of some level of: i) metabolic rate control or depression during warm temperature exposures; and ii) homeostasis/canalization (i.e., absence or low levels of plasticity) in physiological traits that may reflect some sort of buffering mechanism in most of the populations. Nevertheless, our results indicate that elevated temperatures can reduce cardiac function but not metabolic rate in high latitude crabs. The lack of congruence between HR and MR supports the idea that energy metabolism in marine invertebrates cannot be inferred from HR and different conclusions regarding geographic differentiation in energy metabolism can be obtained from both physiological traits. Integrating thermal physiology and species range extent can contribute to a better understanding of the likely effects of climate change on natural populations of marine ectotherms.

Biomineralization changes with food supply confer juvenile scallops (Argopecten purpuratus) resistance to ocean acidification.

Future ocean acidification (OA) will affect physiological traits of marine species, with calcifying species being particularly vulnerable. As OA entails high energy demands, particularly during the rapid juvenile growth phase, food supply may play a key role in the response of marine organisms to OA. We experimentally evaluated the role of food supply in modulating physiological responses and biomineralization processes in juveniles of the Chilean scallop, Argopecten purpuratus, that were exposed to control (pH ~ 8.0) and low pH (pH ~ 7.6) conditions using three food supply treatments (high, intermediate, and low). We found that pH and food levels had additive effects on the physiological response of the juvenile scallops. Metabolic rates, shell growth, net calcification, and ingestion rates increased significantly at low pH conditions, independent of food. These physiological responses increased significantly in organisms exposed to intermediate and high levels of food supply. Hence, food supply seems to play a major role modulating organismal response by providing the energetic means to bolster the physiological response of OA stress. On the contrary, the relative expression of chitin synthase, a functional molecule for biomineralization, increased significantly in scallops exposed to low food supply and low pH, which resulted in a thicker periostracum enriched with chitin polysaccharides. Under reduced food and low pH conditions, the adaptive organismal response was to trade-off growth for the expression of biomineralization molecules and altering of the organic composition of shell periostracum, suggesting that the future performance of these calcifiers will depend on the trajectories of both OA and food supply. Thus, incorporating a suite of traits and multiple stressors in future studies of the adaptive organismal response may provide key insights on OA impacts on marine calcifiers.

Geographic variation in thermal physiological performance of the intertidal crab Petrolisthes violaceus along a latitudinal gradient.

Environmental temperature has profound effects on the biological performance and biogeographical distribution of ectothermic species. Variation of this abiotic factor across geographic gradients is expected to produce physiological differentiation and local adaptation of natural populations depending on their thermal tolerances and physiological sensitivities. Here, we studied geographic variation in whole-organism thermal physiology of seven populations of the porcelain crab Petrolisthes violaceus across a latitudinal gradient of 3000 km, characterized by a cline of thermal conditions. Our study found that populations of P. violaceus show no differences in the limits of their thermal performance curves and demonstrate a negative correlation of their optimal temperatures with latitude. Additionally, our findings show that high-latitude populations of P. violaceus exhibit broader thermal tolerances, which is consistent with the climatic variability hypothesis. Interestingly, under a future scenario of warming oceans, the thermal safety margins of P. violaceus indicate that lower latitude populations can physiologically tolerate the ocean-warming scenarios projected by the IPCC for the end of the twenty-first century.

Environmental correlates of oyster farming in an upwelling system: Implication upon growth, biomass production, shell strength and organic composition.

Comprehending the potential effects of environmental variability on bivalves aquaculture becomes crucial for its sustainability under climate change scenarios, specially in the Humboldt Current System (HCS) where upwelling intensification leading to frequent hypoxia and acidification is expected. In a year-long study, Pacific oysters (Magallana gigas) were monitored at two depths (1.5m, 6.5m) in a bay affected by coastal upwelling. Surface waters exhibited warmer, well-oxygenated conditions and higher chlorophyll-a concentrations, while at depth greater hypoxia and acidification events occur, especially during upwelling. Surface cultured oysters exhibited 60 % larger size and 35% greater weight due to faster growth rate during the initial month of cultivation. The condition index (CI) increases in surface oysters after 10 months, whereas those at the bottom maintain a lower index. Food availability, temperature, and oxygen, correlates with higher growth rates, while pH associates with morphometric variables, indicating that larger oysters tend to develop under higher pH. Increased upwelling generally raises CI, but bottom oysters face stressful conditions such as hypoxia and acidification, resulting in lower performance. However, they acclimate by changing the organic composition of their shells and making them stronger. This study suggests that under intensified upwelling scenario, oysters would grow slowly, resulting in smaller sizes and lower performance, but the challenges may be confronted through complex compensation mechanisms among biomass production and maintenance of the shell structure and function. This poses a significant challenge for the sustainability of the aquaculture industry, emphasizing the need for adaptive strategies to mitigate the effects of climate change.

Differential gene expression analysis in the scallop Argopecten purpuratus exposed to altered pH and temperature conditions in an upwelling-influenced farming area.

Increased carbon dioxide in the atmosphere and its absorption across the ocean surface will alter natural variations in pH and temperature levels, occurring in coastal upwelling ecosystems. The scallop Argopecten purpuratus, one of the most economically important species farmed in northern Chile, has been shown to be vulnerable to these environmental drivers. However, the regulatory responses at the gene-level of scallops to these climate stressors remain almost unknown. Consequently, we used an orthogonal experimental design and RNAseq approach to analyze the acute effects of variability in pH and temperature on gene expression in the muscle tissue of A. purpuratus. In respect to control conditions (pH ~ 8.0/ 14 °C), the influence of low pH (~ 7.7) and temperature (14 °C) induced the activation of several genes associated with apoptotic signaling pathways and protein localization to plasma membrane. Elevated temperature (18 °C) and pH (~8.0) conditions increased the expression of transcripts associated with the activation of muscle contraction, regulation, and sarcomere organization effects on muscle tissue. In scallops exposed to low pH and elevated temperature, the genes expressed were differentially associated with the oxidation-reduction process, signal translation, and positive regulation of GTPase activity. These results indicated that the differentially expressed genes under the experimental conditions tested are mainly related to the mitigation of cellular damage and homeostasis control. Our results add knowledge about the function of the adductor muscle in response to stressors in scallops. Furthermore, these results could help in the identification of molecular biomarkers of stress necessary to be integrated into the aquaculture programs for the mitigation of climate change.

Direct and indirect impacts of ocean acidification and warming on algae-herbivore interactions in intertidal habitats.

Anthropogenically induced global climate change has caused profound impacts in the world ocean. Climate change related stressors, like ocean acidification (OA) and warming (OW) can affect physiological performance of marine species. However, studies evaluating the impacts of these stressors on algae-herbivore interactions have been much more scarce. We approached this issue by assessing the combined impacts of OA and OW on the physiological energetics of the herbivorous snail Tegula atra, and whether this snail is affected indirectly by changes in biochemical composition of the kelp Lessonia spicata, in response to OA and OW. Our results show that OA and OW induce changes in kelp biochemical composition and palatability (organic matter, phenolic content), which in turn affect snails' feeding behaviour and energy balance. Nutritional quality of food plays a key role on grazers' physiological energetics and can define the stability of trophic interactions in rapidly changing environments such as intertidal communities.

Adaptive shell color plasticity during the early ontogeny of an intertidal keystone snail.

We report a mechanism of crypsis present during the vulnerable early post-metamorphic ontogeny (95%) of specimens bearing patterns of shell coloration (dark or light colored) that matched the background coloration provided by patches of Concholepas' most abundant prey (mussels or barnacles respectively). The variation in shell color was positively associated with the color of the most common prey (r = 0.99). In laboratory experiments, shell coloration of C. concholepas depended on the prey-substrate used to induce metamorphosis and for the post-metamorphic rearing. The snail shell color matched the color of the prey offered during rearing. Laboratory manipulation experiments, switching the prey during rearing, showed a corresponding change in snail shell color along the outermost shell edge. As individuals grew and became increasingly indistinguishable from the surrounding background, cryptic individuals had higher survival (71%) than the non cryptic ones (4%) when they were reared in the presence of the predatory crab Acanthocyclus hassleri. These results suggest that the evolution of shell color plasticity during the early ontogeny of C. concholepas, depends on the color of the more abundant of the consumed prey available in the natural habitat where settlement has taken place; this in turn has important consequences for their fitness and survivorship in the presence of visual predators.

Evaluation of remodeling and geometry on the biomechanical properties of nacreous bivalve shells.

Mollusks have developed a broad diversity of shelled structures to protect against challenges imposed by biological interactions(e.g., predation) and constraints (e.g., [Formula: see text]-induced ocean acidification and wave-forces). Although the study of shell biomechanical properties with nacreous microstructure has provided understanding about the role of shell integrity and functionality on mollusk performance and survival, there are no studies, to our knowledge, that delve into the variability of these properties during the mollusk ontogeny, between both shells of bivalves or across the shell length. In this study, using as a model the intertidal mussel Perumytilus purpuratus to obtain, for the first time, the mechanical properties of its shells with nacreous microstructure; we perform uniaxial compression tests oriented in three orthogonal axes corresponding to the orthotropic directions of the shell material behavior (thickness, longitudinal, and transversal). Thus, we evaluated whether the shell material's stress and strain strength and elastic modulus showed differences in mechanical behavior in mussels of different sizes, between valves, and across the shell length. Our results showed that the biomechanical properties of the material building the P. purpuratus shells are symmetrical in both valves and homogeneous across the shell length. However, uniaxial compression tests performed across the shell thickness showed that biomechanical performance depends on the shell size (aging); and that mechanical properties such as the elastic modulus, maximum stress, and strain become degraded during ontogeny. SEM observations evidenced that compression induced a tortuous fracture with a delamination effect on the aragonite mineralogical structure of the shell. Findings suggest that P. purpuratus may become vulnerable to durophagous predators and wave forces in older stages, with implications in mussel beds ecology and biodiversity of intertidal habitats.

Parasitism by metacercariae modulates the morphological, organic and mechanical responses of the shell of an intertidal bivalve to environmental drivers.

Environmental variation alters biological interactions and their ecological and evolutionary consequences. In coastal systems, trematode parasites affect their hosts by disrupting their life-history traits. However, the effects of parasitism could be variable and dependent on the prevailing environmental conditions where the host-parasite interaction occurs. This study compared the effect of a trematode parasite in the family Renicolidae (metacercariae) on the body size and the shell organic and mechanical characteristics of the intertidal mussels Perumytilus purpuratus, inhabiting two environmentally contrasting localities in northern and central Chile (ca. 1600 km apart). Congruent with the environmental gradient along the Chilean coast, higher levels of temperature, salinity and pCO2, and a lower pH characterise the northern locality compared to that of central Chile. In the north, parasitised individuals showed lower body size and shell resistance than non-parasitised individuals, while in central Chile, the opposite pattern was observed. Protein level in the organic matter of the shell was lower in the parasitised hosts than in the non-parasitised ones regardless of the locality. However, an increase in polysaccharide levels was observed in the parasitised individuals from central Chile. These results evidence that body size and shell properties of P. purpuratus vary between local populations and that they respond differently when confronting the parasitism impacts. Considering that the parasite prevalence reaches around 50% in both populations, if parasitism is not included in the analysis, the true response of the host species would be masked by the effects of the parasite, skewing our understanding of how environmental variables will affect marine species. Considering parasitism and identifying its effects on host species faced with environmental drivers is essential to understand and accurately predict the ecological consequences of climate change.

Morphological, physiological and behavioral responses of an intertidal snail, Acanthina monodon (Pallas), to projected ocean acidification and cooling water conditions in upwelling ecosystems.

Ocean acidification (OA) is expected to rise towards the end of the 21st century altering the life history traits in marine organisms. Upwelling systems will not escape OA, but unlike other areas of the ocean, cooling effects are expected to intensify in these systems. Regardless, studies evaluating the combined effects of OA and cooling remain scarce. We addressed this gap using a mesocosm system, where we exposed juveniles of the intertidal muricid snail Acanthina monodon to current and projected pCO2 (500 vs. 1500 ppm) and temperature (15 vs. 10 °C) from the southeast Pacific upwelling system. After 9 weeks of experimental exposure to those conditions, we conducted three estimations of growth (wet weight, shell length and shell peristomal length), in addition to measuring calcification, metabolic and feeding rates and the ability of these organisms to return to the normal upright position after being overturned (self-righting). Growth, feeding and calcification rates increased in projected cooling conditions (10 °C) but were unaffected by pCO2 or the interaction between pCO2 and temperature. Instead, metabolic rates were driven by pCO2, but a significant interaction with temperature suggests that in cooler conditions, metabolic rates will increase when associated with high pCO2 levels. Snail self-righting times were not affected across treatments. These results suggest that colder temperatures projected for this area would drive this species growth, feeding and calcification, and consequently, some of its population biology and productivity. However, the snails may need to compensate for the increase in metabolic rates under the effects of ocean acidification. Although A. monodon ability to adjust to individual or combined stressors will likely account for some of the changes described here, our results point to a complex dynamic to take place in intertidal habitats associated with upwelling systems.

Ocean acidification alters anti-predator responses in a competitive dominant intertidal mussel.

Widespread intertidal mussels are exposed to a variety of natural and anthropogenic stressors. Even so, our understanding of the combined influence of stressors such as predation risk and ocean acidification (OA) on these species remains limited. This study examined the response of the purple mussel (Perumytilus purpuratus), a species distributed along Pacific southeastern rocky shores, to the effects of predation risk and OA. Using a laboratory 2 × 2 cross design, purple mussels were either devoid or exposed to predator cues from the muricid snail Acanthina monodon, while simultaneously exposing them to current (500 ppm) or projected OA conditions (1500 ppm). The response of purple mussels to these factors was assessed using growth, calcification, clearance, and metabolic rates, in addition to byssus production. After 60 d, the presence of predator cues reduced mussel growth in width and length, and in the latter case, OA enhanced this response making the effects of predator cues more severe. Calcification rates were driven by the interaction between the two stressors, whereas clearance rates increased only in response to OA, likely explaining some of the growth results. Mussel byssus production also increased with pCO2 but interacted with predation risk: in the absence of predator cues, byssus production increased with OA. These results suggest that projected levels of OA may alter and in some cases prevail over the natural response of purple mussels to predation risk. Considering the role played by this mussel as a dominant competitor and ecosystem engineer in rocky shores, these results have community-wide implications.

Metabolic rate allometry in intertidal mussels across environmental gradients: The role of coastal carbonate system parameters in mediating the effects of latitude and temperature.

We assess the role of direct and indirect effects of coastal environmental drivers (including the parameters of the carbonate system) on energy expenditure (MR) and body mass (M) of the intertidal mussel, Perumytilus purpuratus, across 10 populations distributed over 2800 km along the Southern Eastern Pacific (SEP) coast. We find biogeographic and local variation in carbonate system variables mediates the effects of latitude and temperature on metabolic rate allometry along the SEP coast. Also, the fitted Piecewise Structural Equation models (PSEM) have greater predictive ability (conditional R2 = 0.95) relative to the allometric scaling model (R2 = 0.35). The largest standardized coefficients for MR and M were determined by the influence of temperature and latitude, followed by pCO2, pH, total alkalinity, and salinity. Thus, physiological diversity of P. purpuratus along the SEP coast emerges as the result of direct and indirect effects of biogeographic and local environmental variables.

Biomechanical Characterization of Scallop Shells Exposed to Ocean Acidification and Warming.

Increased carbon dioxide levels (CO2) in the atmosphere triggered a cascade of physical and chemical changes in the ocean surface. Marine organisms producing carbonate shells are regarded as vulnerable to these physical (warming), and chemical (acidification) changes occurring in the oceans. In the last decade, the aquaculture production of the bivalve scallop Argopecten purpuratus (AP) showed declined trends along the Chilean coast. These negative trends have been ascribed to ecophysiological and biomineralization constraints in shell carbonate production. This work experimentally characterizes the biomechanical response of AP scallop shells subjected to climate change scenarios (acidification and warming) via quasi-static tensile and bending tests. The experimental results indicate the adaptation of mechanical properties to hostile growth scenarios in terms of temperature and water acidification. In addition, the mechanical response of the AP subjected to control climate conditions was analyzed with finite element simulations including an anisotropic elastic constitutive model for a two-fold purpose: Firstly, to calibrate the material model parameters using the tensile test curves in two mutually perpendicular directions (representative of the mechanical behavior of the material). Secondly, to validate this characterization procedure in predicting the material's behavior in two mechanical tests.

Evolutionary drivers of the hump-shaped latitudinal gradient of benthic polychaete species richness along the Southeastern Pacific coast.

Latitudinal diversity gradients (LDG) and their explanatory factors are among the most challenging topics in macroecology and biogeography. Despite of its apparent generality, a growing body of evidence shows that 'anomalous' LDG (i.e., inverse or hump-shaped trends) are common among marine organisms along the Southeastern Pacific (SEP) coast. Here, we evaluate the shape of the LDG of marine benthic polychaetes and its underlying causes using a dataset of 643 species inhabiting the continental shelf (<200 m depth), using latitudinal bands with a spatial resolution of 0.5°, along the SEP (3-56° S). The explanatory value of six oceanographic (Sea Surface Temperature (SST), SST range, salinity, salinity range, primary productivity and shelf area), and one macroecological proxy (median latitudinal range of species) were assessed using a random forest model. The taxonomic structure was used to estimate the degree of niche conservatism of predictor variables and to estimate latitudinal trends in phylogenetic diversity, based on three indices (phylogenetic richness (PDSES), mean pairwise distance (MPDSES), and variation of pairwise distances (VPD)). The LDG exhibits a hump-shaped trend, with a maximum peak of species richness at ca. 42° S, declining towards northern and southern areas of SEP. The latitudinal pattern was also evident in local samples controlled by sampling effort. The random forest model had a high accuracy (pseudo-r2 = 0.95) and showed that the LDG could be explained by four variables (median latitudinal range, SST, salinity, and SST range), yet the functional relationship between species richness and these predictors was variable. A significant degree of phylogenetic conservatism was detected for the median latitudinal range and SST. PDSES increased toward the southern region, whereas VPD showed the opposite trend, both statistically significant. MPDSES has the same trend as PDSES, but it is not significant. Our results reinforce the idea that the south Chile fjord area, particularly the Chiloé region, was likely the evolutionary source of new species of marine polychaetes along SEP, creating a hotspot of diversity. Therefore, in the same way as the canonical LDG shows a decline in diversity while moving away from the tropics; on this case the decline occurs while moving away from Chiloé Island. These results, coupled with a strong phylogenetic signal of the main predictor variables suggest that processes operating mainly at evolutionary timescales govern the LDG.

Plasticity in organic composition maintains biomechanical performance in shells of juvenile scallops exposed to altered temperature and pH conditions.

The exposure to environmental variations in pH and temperature has proven impacts on benthic ectotherms calcifiers, as evidenced by tradeoffs between physiological processes. However, how these stressors affect structure and functionality of mollusk shells has received less attention. Episodic events of upwelling of deep cold and low pH waters are well documented in eastern boundary systems and may be stressful to mollusks, impairing both physiological and biomechanical performance. These events are projected to become more intense, and extensive in time with ongoing global warming. In this study, we evaluate the independent and interactive effects of temperature and pH on the biomineral and biomechanical properties of Argopecten purpuratus scallop shells. Total organic matter in the shell mineral increased under reduced pH (~ 7.7) and control conditions (pH ~ 8.0). The periostracum layer coating the outer shell surface showed increased protein content under low pH conditions but decreasing sulfate and polysaccharides content. Reduced pH negatively impacts shell density and increases the disorder in the orientation of calcite crystals. At elevated temperatures (18 °C), shell microhardness increased. Other biomechanical properties were not affected by pH/temperature treatments. Thus, under a reduction of 0.3 pH units and low temperature, the response of A. purpuratus was a tradeoff among organic compounds (biopolymer plasticity), density, and crystal organization (mineral plasticity) to maintain shell biomechanical performance, while increased temperature ameliorated the impacts on shell hardness. Biopolymer plasticity was associated with ecophysiological performance, indicating that, under the influence of natural fluctuations in pH and temperature, energetic constraints might be critical in modulating the long-term sustainability of this compensatory mechanism.

Microstructure and crystallography of the wall plates of the giant barnacle Austromegabalanus psittacus: a material organized by crystal growth.

In biomineralization, it is essential to know the microstructural and crystallographic organization of natural hard tissues. This knowledge is virtually absent in the case of barnacles. Here, we have examined the crystal morphology and orientation of the wall plates of the giant barnacle Austromegabalanus psittacus by means of optical and electron microscopy, and electron backscatter diffraction. The wall plates are made of calcite grains, which change in morphology from irregular to rhombohedral, except for the radii and alae, where fibrous calcite is produced. Both the grains and fibres arrange into bundles made of crystallographically co-oriented units, which grow onto each other epitaxially. We call these areas crystallographically coherent regions (CCRs). Each CCR elongates and disposes its c-axis perpendicularly or at a high angle to the growth surfaces, whereas the a-axes of adjacent CCRs differ in orientation. In the absence of obvious organic matrices, this pattern of organization is interpreted to be produced by purely crystallographic processes. In particular, due to crystal competition, CCRs orient their fastest growth axes perpendicular to the growth surface. Since each CCR is an aggregate of grains, the fastest growth axis is that along which crystals stack up more rapidly, that is, the crystallographic c-axis in granular calcite. In summary, the material forming the wall plates of the studied barnacles is under very little biological control and the main role of the mantle cells is to provide the construction materials to the growth front.

Origin of the biphase nature and surface roughness of biogenic calcite secreted by the giant barnacle Austromegabalanus psittacus.

The calcite grains forming the wall plates of the giant barnacle Austramegabalanus psittacus have a distinctive surface roughness made of variously sized crystalline nanoprotrusions covered by extremely thin amorphous pellicles. This biphase (crystalline-amorphous) structure also penetrates through the crystal's interiors, forming a web-like structure. Nanoprotrusions very frequently elongate following directions related to the crystallographic structure of calcite, in particular, the <- 441> directions, which are the strongest periodic bond chains (PBCs) in calcite. We propose that the formation of elongated nanoprotrusions happens during the crystallization of calcite from a precursor amorphous calcium carbonate (ACC). This is because biomolecules integrated within the ACC are expelled from such PBCs due to the force of crystallization, with the consequent formation of uninterrupted crystalline nanorods. Expelled biomolecules accumulate in adjacent regions, thereby stabilizing small pellicle-like volumes of ACC. With growth, such pellicles become occluded within the crystal. In summary, the surface roughness of the biomineral surface reflects the complex shape of the crystallization front, and the biphase structure provides evidence for crystallization from an amorphous precursor. The surface roughness is generally explained as resulting from the attachment of ACC particles to the crystal surface, which later crystallised in concordance with the crystal lattice. If this was the case, the nanoprotrusions do not reflect the size and shape of any precursor particle. Accordingly, the particle attachment model for biomineral formation should seek new evidence.

Geographical variability and parasitism on body size, reproduction and shell characteristics of the keyhole limpet Fissurella crassa (Mollusca: Vetigastropoda).

Environmental variation may alter biological interactions and their ecological consequences. For instance, in marine ecosystems hosts and parasites are subject to environmental variability across latitudinal gradients, and their co-evolutionary dynamics may be the result of the interplay with local physical-chemical variables in seawater. Thus, assessing the environmental conditions required for a host in order to improve their survival is essential to understand the host-parasite interaction and dynamics. In this study, we evaluated the impact of parasitism by Proctoeces humboldti on the body size and reproduction of the intertidal keyhole limpet Fissurella crassa collected from three populations spanning ca. 1500 km along the latitudinal gradient of the Chilean coast. In addition, for the first time, we explore whether the effect of parasitism can be extended to changes in the organic composition and mechanical properties of the host shell. Our results show that parasitism prevalence and intensity, and body size of F. crassa increased in central Chile (ca. 33°S). Unlike body size, which was greater in parasitized limpets than in non-parasitized limpets at the three study sites, reproductive performance followed this trend only in central Chile populations, with no differences between parasitized and non-parasitized limpets collected in the northern Chilean (ca. 23°S), and lower in parasitized than non-parasitized individuals from the south-central Chile (ca. 37°S). The organic composition of F. crassa shells showed significant differences between parasite conditions (e.g. polysaccharides and water decreased in parasitized limpets) and across sites (e.g. proteins levels increase in shell of parasitized limpets from central Chile, but decreased at south-central Chile). However, variability in shell mechanical properties (e.g. toughness and elastic module) do not showed significant differences across sites and parasitism condition. These results suggest the interplay of both parasitism and environmental fluctuations upon the reproductive performance and morphology of the host. In addition, our result highlight that the host may also trade-offs reproduction, growth and shell organic composition to maintain the shell functionality (e.g. protection for mechanical forces and durophagous predators).