Microclimate drives intraspecific thermal specialization: conservation perspectives in freshwater habitats.

Endemic and relict species are often confined to ecological refugia or over fragmented distributions, representing priority conservation subjects. Within these sites, the individual population may realize distinct niches to a varying degree of specialization. An emblematic example is provided by freshwater species segregated in thermal-mineral springs, where individuals may face highly diverse microclimates in limited geographic areas. Downscaling the characterization of physiological traits to microclimatic niches becomes pivotal to adopt effective conservation measures in these heterogeneous habitats. Melanopsis etrusca (Brot, 1862) is an endangered relict snail endemic to a small number of thermal-mineral streams in central Italy. Here we describe the thermal tolerance of two populations of M. etrusca inhabiting streams with distinctly different thermal regimes, investigating the extent of physiological and behavioural specialization to such diverse microclimatic niches. The comparison of oxygen consumption rates of a population dwelling in temperate streams, characterized by seasonal temperature fluctuations (12-27°C), with a population experiencing constantly hot water (35-38°C) revealed the absence of any seasonal or geographic effect on metabolic compensation. Conversely, mobility performances were maximized in the population inhabiting the hot stream. Interestingly, here, the snails exhibited emersion behaviour outside the water, triggered by temperatures above 37°C. In the field, individuals of this population are observed inactive on stream banks, conceivably to minimize the metabolic cost that otherwise would be induced by remaining in the hot water. Only a few individuals from the temperate stream exhibited the same behaviour when exposed to elevated temperatures, suggesting the exaptation of a pre-existing trait during the evolutionary process of adaptation to hot waters. The present results provide elements for the best practice in future programmes aimed at reintroducing stocks of threatened species across heterogeneous habitats. Our study further underlines the relevance of downscaling data collection for endangered species conservation in order to recognize microclimatic specializations.

Sea urchin chronicles. The effect of oxygen super-saturation and marine polluted sediments from Bagnoli-Coroglio Bay on different life stages of the sea urchin Paracentrotus lividus.

In marinas and harbours, the accumulation of pollutants in sediments, combined with poor exchange of water with the open sea, poses a major environmental threat. The presence of photosynthetic organisms and the related oxygen production, however, may alleviate the negative effects of environmental contamination on heterotrophic organisms, enhancing their physiological defences. Furthermore, possible transgenerational buffer effects may increase the ability of natural populations to face environmental stress. Here we tested the occurrence of transgenerational effects on larvae of the sea urchin Paracentrotus lividus, whose parents were exposed, during the gametogenesis, to contaminated sediments subject to two temporal patterns of water re-suspension events and normal- (90%) vs. super-saturated (200%) levels of O2. The study site was Bagnoli-Coroglio (Gulf of Naples, southern Tyrrhenian Sea), a historically polluted brownfield and Site of National Interest for which environmental restoration options are currently under exploration. Larvae from different adult populations were significantly, although not linearly, affected by the interaction of all factors to which parents were exposed, at both 24h and 48h post fertilization. Specifically, the exposure of larvae to elutriates from contaminated sediments determined a developmental delay, a reduction in size and an increased percentage of abnormalities in all larval populations independently of their parental exposure. On the contrary, larvae from parents exposed to contaminated sediments, when reared in clean filtered sea water, succeeded in developing until the echinopluteus stage after 48h, with size and abundance comparable to those of larvae from control parents. Pre-exposure of parents to contaminated sediments did not successfully buffer the negative effects of elutriates on their offspring, and no positive effects of 'super-saturated' levels of O2 in response to contaminants were observed, suggesting that the Bagnoli-Coroglio area is currently not suitable for the re-stocking or re-introduction of this species.

Texture features explain the susceptibility of grapevine cultivars to Drosophila suzukii (Diptera: Drosophilidae) infestation in ripening and drying grapes.

Grapevine is a well-known host plant of the invasive pest Drosophila suzukii, but its susceptibility to pest oviposition and development greatly depends on the cultivar. To address environmental sustainability during viticultural zoning planning, new vineyard plantation and Integrated Pest Management programmes, it is essential to take pest pressure and cultivar susceptibility into account. To determine the different grapevine cultivars susceptibility to D. suzukii, we tested twelve widely spread cultivars during the ripening period. We also tested three cultivars during the drying period for raisin wine production. The infestation and emergence rates were consequently related to chemical and texture features of the berries to explain the role of skin and pulp characteristics in determining the nature of the susceptibility. Our results showed that susceptibility to D. suzukii infestation varies across cultivars. On ripening grapes, infestation is primarily influenced by skin and pulp firmness, elasticity and consistency. Suitability for egg development resulted mainly related to skin and pulp deformation. In a drying loft, infestation may also occur in relation to skin and pulp consistency. Lastly, we discuss the practical implication of the underestimated role of berry texture in D. suzukii oviposition and emergence success, in both ripening and drying grapes.

Oxygen supersaturation protects coastal marine fauna from ocean warming.

Ocean warming affects the life history and fitness of marine organisms by, among others, increasing animal metabolism and reducing oxygen availability. In coastal habitats, animals live in close association with photosynthetic organisms whose oxygen supply supports metabolic demands and may compensate for acute warming. Using a unique high-frequency monitoring dataset, we show that oxygen supersaturation resulting from photosynthesis closely parallels sea temperature rise during diel cycles in Red Sea coastal habitats. We experimentally demonstrate that oxygen supersaturation extends the survival to more extreme temperatures of six species from four phyla. We clarify the mechanistic basis of the extended thermal tolerance by showing that hyperoxia fulfills the increased metabolic demand at high temperatures. By modeling 1 year of water temperatures and oxygen concentrations, we predict that oxygen supersaturation from photosynthetic activity invariably fuels peak animal metabolic demand, representing an underestimated factor of resistance and resilience to ocean warming in ectotherms.

Ocean acidification and elevated temperature negatively affect recruitment, oxygen consumption and calcification of the reef-building Dendropoma cristatum early life stages: Evidence from a manipulative field study.

Expected temperature rise and seawater pH decrease may affect marine organism fitness. By a transplant experiment involving air-temperature manipulation along a natural CO2 gradient, we investigated the effects of high pCO2 (~1100 µatm) and elevated temperature (up to +2 °C than ambient conditions) on the reproductive success, recruitment, growth, shell chemical composition and oxygen consumption of the early life stages of the intertidal reef-building vermetid Dendropoma cristatum. Reproductive success was predominantly affected by temperature increase, with encapsulated embryos exhibiting higher survival in control than elevated temperature conditions, which were in turn unaffected by altered seawater pH levels. Decreasing pH (alone or in combination with temperature) significantly affected the shell growth and shell chemical composition of both embryos and recruits. Elevated temperatures along with lower pH led to decreases of ~30% oxygen consumption and ~60% recruitment. Our results suggest that the early life stages of the reef-builder D. cristatum are highly sensitive to expected environmental change, with major consequences on the intertidal vermetid reefs they build and indirectly on the high biodiversity levels they support.

Winter temperature predicts prolonged diapause in pine processionary moth species across their geographic range.

Prolonged diapause occurs in a number of insects and is interpreted as a way to evade adverse conditions. The winter pine processionary moths (Thaumetopoea pityocampa and Th. wilkinsoni) are important pests of pines and cedars in the Mediterranean region. They are typically univoltine, with larvae feeding across the winter, pupating in spring in the soil and emerging as adults in summer. Pupae may, however, enter a prolonged diapause with adults emerging one or more years later. We tested the effect of variation in winter temperature on the incidence of prolonged diapause, using a total of 64 individual datasets related to insect cohorts over the period 1964-2015 for 36 sites in seven countries, covering most of the geographic range of both species. We found high variation in prolonged diapause incidence over their ranges. At both lower and upper ends of the thermal range in winter, prolonged diapause tended to be higher than at intermediate temperatures. Prolonged diapause may represent a risk-spreading strategy to mitigate climate uncertainty, although it may increase individual mortality because of a longer exposure to mortality factors such as predation, parasitism, diseases or energy depletion. Climate change, and in particular the increase of winter temperature, may reduce the incidence of prolonged diapause in colder regions whereas it may increase it in warmer ones, with consequences for population dynamics.

Temperature-induced phenotypic plasticity in the ovipositor of the invasive species Drosophila suzukii.

Drosophila suzukii (Matsumura, 1931) is a highly successful invasive dipteran which represents a serious threat for global fruit industry. Among other adaptive traits, D. suzukii owes its success to the derived morphological features of its ovipositor, which allows the insect to exploit the exclusive ecological niche of fresh fruit, thus avoiding competition with other closely related species. With the aim of investigating temperature-induced phenotypic plasticity of D. suzukii ovipositor, we reared this insect in four different laboratory conditions, represented by the combination of two developmental temperatures and two diet regimes for the larvae. We recorded the effects of these two factors on ovipositor size and shape and overall body size through a combination of distance-based and geometric morphometric analyses. Results showed that insects attain the largest body sizes at lower temperature, whereas the diet does not determine significant difference in size. However, the effect on size of the two factors is less pronounced in the ovipositor, which shows a negative allometry with respect to body size in all treatments. At higher temperature, ovipositor shape tends also to co-vary with its own size. Neither temperature nor diet have significant effect on ovipositor bilateral fluctuating asymmetry. These results confirm the hypothesis that in D. suzukii the toughened valve of the ovipositor are subjected to effective morpho-functional constraints, while probably being under strong selection by reason of their mechanical role.

Effects of ocean acidification on embryonic respiration and development of a temperate wrasse living along a natural CO2 gradient.

Volcanic CO2 seeps provide opportunities to investigate the effects of ocean acidification on organisms in the wild. To understand the influence of increasing CO2 concentrations on the metabolic rate (oxygen consumption) and the development of ocellated wrasse early life stages, we ran two field experiments, collecting embryos from nesting sites with different partial pressures of CO2 [pCO2; ambient (∼400 µatm) and high (800-1000 µatm)] and reciprocally transplanting embryos from ambient- to high-CO2 sites for 30 h. Ocellated wrasse offspring brooded in different CO2 conditions had similar responses, but after transplanting portions of nests to the high-CO2 site, embryos from parents that spawned in ambient conditions had higher metabolic rates. Although metabolic phenotypic plasticity may show a positive response to high CO2, it often comes at a cost, in this case as a smaller size at hatching. This can have adverse effects because smaller larvae often exhibit a lower survival in the wild. However, the adverse effects of increased CO2 on metabolism and development did not occur when embryos from the high-CO2 nesting site were exposed to ambient conditions, suggesting that offspring from the high-CO2 nesting site could be resilient to a wider range of pCO2 values than those belonging to the site with present-day pCO2 levels. Our study identifies a crucial need to increase the number of studies dealing with these processes under global change trajectories and to expand these to naturally high-CO2 environments, in order to assess further the adaptive plasticity mechanism that encompasses non-genetic inheritance (epigenetics) through parental exposure and other downstream consequences, such as survival of larvae.

Concurrent environmental stressors and jellyfish stings impair caged European sea bass (Dicentrarchus labrax) physiological performances.

The increasing frequency of jellyfish outbreaks in coastal areas has led to multiple ecological and socio-economic issues, including mass mortalities of farmed fish. We investigated the sensitivity of the European sea bass (Dicentrarchus labrax), a widely cultured fish in the Mediterranean Sea, to the combined stressors of temperature, hypoxia and stings from the jellyfish Pelagia noctiluca, through measurement of oxygen consumption rates (MO2), critical oxygen levels (PO2crit), and histological analysis of tissue damage. Higher levels of MO2, PO2crit and gill damage in treated fish demonstrated that the synergy of environmental and biotic stressors dramatically impair farmed fish metabolic performances and increase their health vulnerability. As a corollary, in the current scenario of ocean warming, these findings suggest that the combined effects of recurrent hypoxic events and jellyfish blooms in coastal areas might also threaten wild fish populations.

The importance of thermal history: costs and benefits of heat exposure in a tropical, rocky shore oyster.

Although thermal performance is widely recognised to be pivotal in determining species' distributions, assessment of this performance is often based on laboratory-acclimated individuals, neglecting their proximate thermal history. The thermal history of a species sums the evolutionary history and, importantly, the thermal events recently experienced by individuals, including short-term acclimation to environmental variations. Thermal history is perhaps of greatest importance for species inhabiting thermally challenging environments and therefore assumed to be living close to their thermal limits, such as in the tropics. To test the importance of thermal history, the responses of the tropical oyster Isognomon nucleus to short-term differences in thermal environments were investigated. Critical and lethal temperatures and oxygen consumption were improved in oysters that previously experienced elevated air temperatures, and were associated with an enhanced heat shock response, indicating that recent thermal history affects physiological performance as well as inducing short-term acclimation to acute conditions. These responses were, however, associated with trade-offs in feeding activity, with oysters that experienced elevated temperatures showing reduced energy gain. Recent thermal history, therefore, seems to rapidly invoke physiological mechanisms that enhance survival of short-term thermal challenge but also longer term climatic changes and consequently needs to be incorporated into assessments of species' thermal performances.

The trade-off between heat tolerance and metabolic cost drives the bimodal life strategy at the air-water interface.

The principle of oxygen and capacity limitation of thermal tolerance in ectotherms suggests that the long-term upper limits of an organism's thermal niche are equivalent to the upper limits of the organism's functional capacity for oxygen provision to tissues. Air-breathing ectotherms show wider thermal tolerances, since they can take advantage of the higher availability of oxygen in air than in water. Bimodal species move from aquatic to aerial media and switch between habitats in response to environmental variations such as cyclical or anomalous temperature fluctuations. Here we tested the prediction that bimodal species cope better with thermal stress than truly aquatic species using the crab Pachygrapsus marmoratus as a model species. When in water, oxygen consumption rates of P. marmoratus acutely rise during warming. Beyond a temperature threshold of 23 °C the crab's aerobic metabolism in air remains lower than in water. In parallel, the haemolymph oxygen partial pressure of submerged animals progressive decreases during warming, while it remains low but constant during emersion. Our results demonstrate the ability of a bimodal breathing ectotherm to extend its thermal tolerance during air-breathing, suggesting that there are temperature-related physiological benefits during the evolution of the bimodal life style.

Can respiratory physiology predict thermal niches?

Predicting species responses to global warming is the holy grail of climate change science. As temperature directly affects physiological rates, it is clear that a mechanistic understanding of species vulnerability should be grounded in organismal physiology. Here, we review what respiratory physiology can offer the field of thermal ecology, showcasing different perspectives on how respiratory physiology can help explain thermal niches. In water, maintaining adequate oxygen delivery to fuel the higher metabolic rates under warming conditions can become the weakest link, setting thermal tolerance limits. This has repercussions for growth and scaling of metabolic rate. On land, water loss is more likely to become problematic as long as O2 delivery and pH balance can be maintained, potentially constraining species in their normal activity. Therefore, high temperatures need not be lethal, but can still affect the energy intake of an animal, with concomitant consequences for long-term fitness. While respiratory challenges and adaptive responses are diverse, there are clear recurring elements such as oxygen uptake, CO2 excretion, and water homeostasis. We show that respiratory physiology has much to offer the field of thermal ecology and call for an integrative, multivariate view incorporating respiratory challenges, thermal responses, and energetic consequences. Fruitful areas for future research are highlighted.

Improved heat tolerance in air drives the recurrent evolution of air-breathing.

The transition to air-breathing by formerly aquatic species has occurred repeatedly and independently in fish, crabs and other animal phyla, but the proximate drivers of this key innovation remain a long-standing puzzle in evolutionary biology. Most studies attribute the onset of air-breathing to the repeated occurrence of aquatic hypoxia; however, this hypothesis leaves the current geographical distribution of the 300 genera of air-breathing crabs unexplained. Here, we show that their occurrence is mainly related to high environmental temperatures in the tropics. We also demonstrate in an amphibious crab that the reduced cost of oxygen supply in air extends aerobic performance to higher temperatures and thus widens the animal's thermal niche. These findings suggest that high water temperature as a driver consistently explains the numerous times air-breathing has evolved. The data also indicate a central role for oxygen- and capacity-limited thermal tolerance not only in shaping sensitivity to current climate change but also in underpinning the climate-dependent evolution of animals, in this case the evolution of air-breathing.

A role for haemolymph oxygen capacity in heat tolerance of eurythermal crabs.

Heat tolerance in aquatic ectotherms is constrained by a mismatch, occurring at high temperatures, between oxygen delivery and demand which compromises the maintenance of aerobic scope. The present study analyses how the wide thermal tolerance range of an eurythermal model species, the green crab Carcinus maenas is supported and limited by its ability to sustain efficient oxygen transport to tissues. Similar to other eurytherms, C. maenas sustains naturally occurring acute warming events through the integrated response of circulatory and respiratory systems. The response of C. maenas to warming can be characterized by two phases. During initial warming, oxygen consumption and heart rate increase, while stroke volume and haemolymph oxygen partial pressure decrease. During further warming, dissolved oxygen levels in the venous compartment decrease below the threshold of full haemocyanin oxygen saturation. The progressive release of haemocyanin bound oxygen with further warming follows an exponential pattern, thereby saving energy in oxygen transport and causing an associated leveling off of metabolic rate. According to the concept of oxygen and capacity limited thermal tolerance (OCLTT), this indicates that the thermal tolerance window is widened by the increasing contribution of haemocyanin oxygen transport and associated energy savings in cardiocirculation. Haemocyanin bound oxygen sustains cardiac performance to cover the temperature range experienced by C. maenas in the field. To our knowledge this is the first study providing evidence of a relationship between thermal tolerance and blood (haemolymph) oxygen transport in a eurythermal invertebrate.

Climate change reduces offspring fitness in littoral spawners: a study integrating organismic response and long-term time-series.

Integrating long-term ecological observations with experimental findings on species response and tolerance to environmental stress supports an understanding of climate effects on population dynamics. Here, we combine the two approaches, laboratory experiments and analysis of multi-decadal time-series, to understand the consequences of climate anomalies and ongoing change for the population dynamics of a eurythermal littoral species, Carcinus aestuarii. For the generation of cause and effect hypotheses we investigated the thermal response of crab embryos at four developmental stages. We first measured metabolic rate variations in embryos following acute warming (16-24 °C) and after incubation at 20 and 24 °C for limited periods. All experiments consistently revealed differential thermal responses depending on the developmental stage. Temperature-induced changes in metabolic activity of early embryonic stages of blastula and gastrula suggested the onset of abnormal development. In contrast, later developmental stages, characterized by tissue and organ differentiation, were marginally affected by temperature anomalies, indicating enhanced resilience to thermal stress. Then, we extended these findings to a larger, population scale, by analyzing a time-series of C. aestuarii landings in the Venice lagoon from 1945 to 2010 (ripe crabs were recorded separately) in relation to temperature. Landings and extreme climatic events showed marked long-term and short-term variations. We found negative relationships between landings and thermal stress indices on both timescales, with time lags consistent with an impact on crab early life stages. When quantitatively evaluating the influence of thermal stress on population dynamics, we found that it has a comparable effect to that of the biomass of spawners. This work provides strong evidence that physiological responses to climatic anomalies translate into population-level changes and that apparently tolerant species may be impacted before the ontogeny of eurythermy. These ontogenetic bottlenecks markedly shape population dynamics and require study to assess the effects of global change.

Coping with cyclic oxygen availability: evolutionary aspects.

Both the gradual rise in atmospheric oxygen over the Proterozoic Eon as well as episodic fluctuations in oxygen over several million-year time spans during the Phanerozoic Era, have arguably exerted strong selective forces on cellular and organismic respiratory specialization and evolution. The rise in atmospheric oxygen, some 2 billion years after the origin of life, dramatically altered cell biology and set the stage for the appearance of multicelluar life forms in the Vendian (Ediacaran) Period of the Neoproterozoic Era. Over much of the Paleozoic, the level of oxygen in the atmosphere was near the present atmospheric level (21%). In the Late Paleozoic, however, there were extended times during which the level of atmospheric oxygen was either markedly lower or markedly higher than 21%. That these Paleozoic shifts in atmospheric oxygen affected the biota is suggested by the correlations between: (1) Reduced oxygen and the occurrences of extinctions, a lowered biodiversity and shifts in phyletic succession, and (2) During hyperoxia, the corresponding occurrence of phenomena such as arthropod gigantism, the origin of insect flight, and the evolution of vertebrate terrestriality. Basic similarities in features of adaptation to hyopoxia, manifest in living organisms at levels ranging from genetic and cellular to physiological and behavioral, suggest the common and early origin of a suite of adaptive mechanisms responsive to fluctuations in ambient oxygen. Comparative integrative approaches addressing the molecular bases of phenotypic adjustments to cyclic oxygen fluctuation provide broad insight into the incremental steps leading to the early evolution of homeostatic respiratory mechanisms and to the specialization of organismic respiratory function.

Quaternary structure and functional properties of Penaeus monodon hemocyanin.

The hemocyanin of the tiger shrimp, Penaeus monodon, was investigated with respect to stability and oxygen binding. While hexamers occur as a major component, dodecamers and traces of higher aggregates are also found. Both the hexamers and dodecamers were found to be extremely stable against dissociation at high pH, independently of the presence of calcium ions, in contrast to the known crustacean hemocyanins. This could be caused by only a few additional noncovalent interactions between amino acids located at the subunit-subunit interfaces. Based on X-ray structures and sequence alignments of related hemocyanins, the particular amino acids are identified. At all pH values, the p50 and Bohr coefficients of the hexamers are twice as high as those of dodecamers. While the oxygen binding of hexamers from crustaceans can normally be described by a simple two-state model, an additional conformational state is needed to describe the oxygen-binding behaviour of Penaeus monodon hemocyanin within the pH range of 7.0 to 8.5. The dodecamers bind oxygen according to the nested Monod-Whyman-Changeaux (MWC) model, as observed for the same aggregation states of other hemocyanins. The oxygen-binding properties of both the hexameric and dodecameric hemocyanins guarantee an efficient supply of the animal with oxygen, with respect to the ratio between their concentrations. It seems that under normoxic conditions, hexamers play the major role. Under hypoxic conditions, the hexamers are expected not to be completely loaded with oxygen. Here, the dodecamers are supposed to be responsible for the oxygen supply.