Time course and magnitude of synthesis of heat-shock proteins in congeneric marine snails (Genus tegula) from different tidal heights

The time course and magnitude of the heat-shock response in relation to severity of thermal stress are important, yet poorly understood, aspects of thermotolerance. We examined patterns of protein synthesis in congeneric marine snails (genus Tegula) that occur at different heights along the subtidal to intertidal gradient after a thermal exposure (30 degrees C for 2.5 h, followed by 50 h recovery at 13 degrees C) that induced the heat-shock response. We monitored the kinetics and magnitudes of protein synthesis by quantifying incorporation of 35S-labeled methionine and cysteine into newly synthesized proteins and observed synthesis of putative heat-shock proteins (hsp's) of size classes 90, 77, 70, and 38 kDa. In the low- to mid-intertidal species, Tegula funebralis, whose body temperature frequently exceeds 30 degrees C during emersion, synthesis of hsp's commenced immediately after heat stress, reached maximal levels 1-3 h into recovery, and returned to prestress levels by 6 h, except for hsp90 (14 h). In contrast, in the low-intertidal to subtidal species, Tegula brunnea, for which 2.5 h at 30 degrees C represents a near lethal heat stress, synthesis of hsp's commenced 2-14 h after heat stress; reached maximal levels after 15-30 h, which exceeded magnitudes of synthesis in T. funebralis; and returned to prestress levels in the case of hsp90 (50 h) and hsp77 (30 h) but not in the case of hsp70 and hsp38. Exposures to 30 degrees C under aerial (emersion) and aquatic (immersion) conditions resulted in differences in hsp synthesis in T. brunnea but not in T. funebralis. The different time courses and magnitudes of hsp synthesis in these congeners suggest that the vertical limits of their distributions may be set in part by thermal stress.     

Heat-shock protein 70 (Hsp70) as a biochemical stress indicator: an experimental field test in two congeneric intertidal gastropods (genus: Tegula)

Although previous studies have demonstrated that heat-shock protein 70 (Hsp70) can be induced by environmental stress, little is known about natural variation in this response over short time scales. We examined how Hsp70 levels varied over days to weeks in two intertidal snail species of the genus Tegula: Sampling was conducted both under naturally changing environmental conditions and in different vertical zones on a rocky shore. The subtidal to low-intertidal T. brunnea was transplanted into shaded and unshaded mid-intertidal cages to assess temporal variation in Hsps under conditions of increased stress. For comparison, the low to mid-intertidal T. funebralis was transplanted into mid-intertidal cages, within this species' natural zone of occurrence. Snails were sampled every 3 to 4 days for one month, and endogenous levels of two Hsp70-kDa family members (Hsp72 and Hsp74) were quantified using solid-phase immunochemistry. Following periods of midday low tides, levels of Hsps increased greatly in transplanted T. brunnea but not in T. funebralis. Levels of Hsps increased less in T. brunnea transplanted to shaded cages than to unshaded cages, suggesting that prolonged emersion and reduction in feeding time per se are factors that are only mildly stressful. Upregulated levels of Hsps returned to base levels within days. In unmanipulated snails collected from their natural zones, Hsp levels showed little change with thermal variation, indicating that these species did not experience thermally stressful conditions during this study. However, under common conditions in the mid-intertidal zone, Hsp70 levels reflected the different thermal sensitivities of the physiological systems of these two species.     

Antipredator behaviour mediated by chemical cues: the role of conspecific alarm signalling and predator labelling in the avoidance response of a marine gastropod

Gastropods represent a challenge in the understanding of alarm signalling. We studied predator avoidance (climbing behaviour) of the marine snail Tegula funebralis in laboratory experiments. Snails were exposed to crude extract of conspecifics, and to water conditioned by actively feeding or non-feeding predatory crabs. Crabs had previously been maintained on different diets, and were accordingly labelled by chemical cues of various origins. Tegula -extract alone released climbing behaviour in May, but not in June. However, during both these months, snails responded to chemical cues from crabs that were actively feeding on Tegula . Crabs labelled by Tegula -diet, and actively feeding on Tegula , also caused more climbing responses compared to crabs labelled by other diets. Chemical cues derived from crabs actively feeding on another snail species, or from non-feeding crabs, did not induce snail climbing no matter the previous feeding history of the predators. When snails received Tegula -extract combined with water conditioned with a non-feeding, Tegula -labelled crab, no climbing occurred. However, when the non-feeding, Tegula -labelled crab was present in the solution of Tegula -extract, moderate climbing responses were obtained. The results imply that climbing responses of T. funebralis are in general caused by the action of a two-component system. This system seems to be a mixture of chemical cues leaking from the tissue of conspecifics when being eaten, and latent conspecific chemicals that are modified in crabs and presumably released with the urine of chemically labelled predators. The modified chemical labels appear to be fully released by crabs when feeding, and moderately released when detecting food. The responses obtained in May with crude extract alone may result from a seasonal change in alarm signalling properties, or a change in behavioural responsiveness of snails exposed to a variable predator regime.     

Non-climatic thermal adaptation: implications for species' responses to climate warming

There is considerable interest in understanding how ectothermic animals may physiologically and behaviourally buffer the effects of climate warming. Much less consideration is being given to how organisms might adapt to non-climatic heat sources in ways that could confound predictions for responses of species and communities to climate warming. Although adaptation to non-climatic heat sources (solar and geothermal) seems likely in some marine species, climate warming predictions for marine ectotherms are largely based on adaptation to climatically relevant heat sources (air or surface sea water temperature). Here, we show that non-climatic solar heating underlies thermal resistance adaptation in a rocky–eulittoral-fringe snail. Comparisons of the maximum temperatures of the air, the snail''s body and the rock substratum with solar irradiance and physiological performance show that the highest body temperature is primarily controlled by solar heating and re-radiation, and that the snail''s upper lethal temperature exceeds the highest climatically relevant regional air temperature by approximately 22°C. Non-climatic thermal adaptation probably features widely among marine and terrestrial ectotherms and because it could enable species to tolerate climatic rises in air temperature, it deserves more consideration in general and for inclusion into climate warming models.     

Altered kelp (Laminariales) phlorotannins and growth under elevated carbon dioxide and ultraviolet-B treatments can influence associated intertidal food webs

Due to the importance of brown algae, such as kelp (Laminariales, Phaeophyta), within most cool nearshore environments, any direct responses of kelp to multiple global changes could alter the integrity of future coastal marine systems. Fifty-five-day manipulation of carbon dioxide (CO₂) and ultraviolet light (UVB) within outdoor sea-tanks, approximating past, present and two predicted future levels, examined the direct influences on Saccharina latissima (= Laminaria saccharina ) and Nereocystis luetkeana development and biochemistry, as well as the indirect influences on a marine herbivore ( Tegula funebralis ; Gastropoda, Mollusca) and on naturally occurring intertidal detritivores. Kelp species displayed variable directional (negative and positive growth) and scale responses to CO₂ and UVB manipulations, which was influenced by interactions. Kelp phlorotannin (phenolic) production in blade tissues was induced by elevated UVB levels, and especially enhanced (additively) by elevated CO₂, further suggesting that some actively growing kelp species are carbon limited in typical nearshore environments. Negative indirect effects upon detritivore consumers fed CO₂-manipulated kelp blade tissues were detected, however, no statistical relationships existed among UVB-treated tissues, and test herbivores did not distinguish between phlorotannin-altered CO₂: UVB-treated kelp blade tissues. Results suggest that past and future conditions differentially benefit these kelp species, which implies a potential for shifts in species abundance and community composition. Higher CO₂ conditions can indirectly impede marine decay processes delaying access to recycled trace nutrients, which may be disruptive to the seasonal regrowth of algae and/or higher trophic levels of nearshore ecosystems.     

Vitelline-coat lysin: Comparison of lysins among three species in the genus Tegula

Vitelline-coat lysins from two species of marine mollusc of the genus Tegula, Tegula lischkei and Tegula sp., were purified and their properties compared with those of Tegula pfeifferi. Cross-reaction tests among these three species proved that the lysin action on the vitelline coat was species specific. Each of the lysins from these three species is a single, basic polypeptide with a molecular weight of 16000-17000 and an isoelectric point of pH 10.5. All of them share a common antibody recognition site(s) for the anti-T. pfeifferi lysin antibody. Their amino acid sequences were analyzed using intact lysins and peptides separated by reverse phase high-performance liquid chromatography after V8 proteinase digestion. The amino acid sequences of the N-terminus (Nos 1-66) from the three species showed 98% homology, and those of the C-terminus (Nos 91-118) showed 89% homology. It was concluded that the species specificity of the vitelline coat lysin appears to be determined by a sequence of 24 residues (Nos 67-90) in the middle region of the molecule.     

Positive selection and propeptide repeats promote rapid interspecific divergence of a gastropod sperm protein

Male-specific proteins have increasingly been reported as targets of positive selection and are of special interest because of the role they may play in the evolution of reproductive isolation. We report the rapid interspecific divergence of cDNA encoding a major acrosomal protein of unknown function (TMAP) of sperm from five species of teguline gastropods. A mitochondrial DNA clock (calibrated by congeneric species divided by the Isthmus of Panama) estimates that these five species diverged 2-10 MYA. Inferred amino acid sequences reveal a propeptide that has diverged rapidly between species. The mature protein has diverged faster still due to high nonsynonymous substitution rates (> 25 nonsynonymous substitutions per site per 10(9) years). cDNA encoding the mature protein (89-100 residues) shows evidence of positive selection (Dn/Ds > 1) for 4 of 10 pairwise species comparisons. cDNA and predicted secondary-structure comparisons suggest that TMAP is neither orthologous nor paralogous to abalone lysin, and thus marks a second, phylogenetically independent, protein subject to strong positive selection in free-spawning marine gastropods. In addition, an internal repeat in one species (Tegula aureotincta) produces a duplicated cleavage site which results in two alternatively processed mature proteins differing by nine amino acid residues. Such alternative processing may provide a mechanism for introducing novel amino acid sequence variation at the amino-termini of proteins. Highly divergent TMAP N-termini from two other tegulines (Tegula regina and Norrisia norrisii) may have originated by such a mechanism.     

The biology and ecology of narrow endemic and widespread plants: a comparative study of trait variation in 20 congeneric pairs

The objective of this study is to examine whether habitat, herbivory and traits related to resource acquisition, resource conservation, reproduction and dispersal differ between narrow endemic plant species and their widespread congeners. We undertook pairwise contrasts of 25 ecological characteristics and biological traits in 20 congeneric pairs of narrow endemic and widespread plant species in the French Mediterranean region. Within each pair, the two species had the same life-form, pollination mode and dispersal mode. Endemic species differed significantly from widespread congeners for a number of attributes. Endemic species occur in habitats on steeper slopes, with higher rock cover and in lower and more open vegetation than their widespread congeners. Endemic species are significantly smaller than widespread species, but show no differences in traits related to resource acquisition (specific leaf area, leaf nitrogen content, maximum photosynthetic rate) or resource conservation (leaf dry matter content). After accounting for their smaller stature, endemic species produce fewer and smaller flowers with less stigma-anther separation and lower pollen/ovule ratios and produce fewer seeds per plant than their widespread congeners. No consistent variation in seed mass and propagule structure was found between congeneric species. Herbivory levels did not differ between congeneric species. Ecological characteristics, notably the occupation of rocky habitats with low aboveground competition, may thus have played an important role in the differentiation of narrow endemic species in the Western Mediterranean. Morphological and ecophysiological traits of narrow endemic species indicate that they are not more stress-tolerant than their widespread congeners. Lower investment in pollen transfer and seed production suggest that local persistence is a key feature of the population ecology of narrow endemic species.     

Heat transfer in a microvascular network: the effect of heart rate on heating and cooling in reptiles (Pogona barbata and Varanus varius)

Thermally-induced changes in heart rate and blood flow in reptiles are believed to be of selective advantage by allowing animal to exert some control over rates of heating and cooling. This notion has become one of the principal paradigms in reptilian thermal physiology. However, the functional significance of changes in heart rate is unclear, because the effect of heart rate and blood flow on total animal heat transfer is not known. I used heat transfer theory to determine the importance of heat transfer by blood flow relative to conduction. I validated theoretical predictions by comparing them with field data from two species of lizard, bearded dragons (Pogona barbata) and lace monitors (Varanus varius). Heart rates measured in free-ranging lizards in the field were significantly higher during heating than during cooling, and heart rates decreased with body mass. Convective heat transfer by blood flow increased with heart rate. Rates of heat transfer by both blood flow and conduction decreased with mass, but the mass scaling exponents were different. Hence, rate of conductive heat transfer decreased more rapidly with increasing mass than did heat transfer by blood flow, so that the relative importance of blood flow in total animal heat transfer increased with mass. The functional significance of changes in heart rate and, hence, rates of heat transfer, in response to heating and cooling in lizards was quantified. For example, by increasing heart rate when entering a heating environment in the morning, and decreasing heart rate when the environment cools in the evening a Pogona can spend up to 44 min longer per day with body temperature within its preferred range. It was concluded that changes in heart rate in response to heating and cooling confer a selective advantage at least on reptiles of mass similar to that of the study animals (0. 21-5.6 kg).     

Critical thermal maximum of Amur sleeper <Emphasis Type="Italic">Perccottus glenii</Emphasis> in different seasons of the year

Seasonal differences in the critical thermal maximum are revealed in Amur sleeper Perccottus glenii at the age of 1+ and 2 years. Maximum values of the critical thermal maximum determined by two criteria (rotation of the body and immobile gill covers) are registered in the summer, and minimum values of this parameter are observed in the winter. Maximum thermal resistance is registered at a low water heating rate (0.2–0.4°C/h), and minimum resistance is reported for a heating rate 5.0–10.5°C/h. The difference between the values of the critical thermal maximum assessed by two criteria reaches 1–2°C. Features of behavior of Amur sleeper are described at different heating rates. Based on the characteristics of thermal adaptation (critical thermal maximum and optimal temperature for growth), Amur sleeper is similar to warm water eurythermal species (goldfish Carassius auratus and carp Cyprinus carpio).     

Male sterility at extreme temperatures: a significant but neglected phenomenon for understanding Drosophila climatic adaptations

The thermal range for viability is quite variable among Drosophila species and it has long been known that these variations are correlated with geographic distribution: temperate species are on average more cold tolerant but more heat sensitive than tropical species. At both ends of their viability range, sterile males have been observed in all species investigated so far. This symmetrical phenomenon restricts the temperature limits within which permanent cultures can be kept in the laboratory. Thermal heat sterility thresholds are very variable across species from 23 degrees C in heat sensitive species up to 31 degrees C in heat tolerant species. In Drosophila melanogaster, genetic variations are observed among geographic populations. Tropical populations are more tolerant to heat induced sterility and recover more rapidly than temperate ones. A genetic analysis revealed that about 50% of the difference observed between natural populations was due to the Y chromosome. Natural populations have not reached a selection limit, however: thermal tolerance was still increased by keeping strains at a high temperature, close to the sterility threshold. On the low temperature side, a symmetrical reverse phenomenon seems to exist: temperate populations are more tolerant to cold than tropical ones. Compared to Mammals, drosophilids exhibit two major differences: first, male sterility occurs not only at high temperature, but also at a low temperature; second, sterility thresholds are not evolutionarily constrained, but highly variable. Altogether, significant and sometimes major genetic variations have been observed between species, between geographic races of the same species, and even between strains kept in the laboratory under different thermal regimes. In each case, it is easily argued that the observed variations correspond to adaptations to climatic conditions, and that male sterility is a significant component of fitness and a target of natural selection.     

Preference versus performance: body temperature of the intertidal snail Chlorostoma funebralis

Evolutionary theory predicts that, in variable environments, it is advantageous for ectothermic organisms to prefer a body temperature slightly below the physiological optimum. This theory works well for many terrestrial organisms but has not been tested for animals inhabiting the hypervariable physical environment of intertidal shores. In laboratory experiments, we allowed the intertidal snail Chlorostoma funebralis to position itself on a temperature gradient, then measured its thermal preference and determined an index of how its performance varied with temperature. Snails performed a biased random walk along the temperature gradient, which, contrary to expectations, caused them to aggregate where body temperature was 15 to 17 °C below their temperature of optimum performance and near the species' lower thermal limit. This "cold-biased" behavioral response may guide snails to refuges in shaded cracks and crevices, but potentially precludes C. funebralis from taking full advantage of its physiological capabilities.     

Thermal niche predicts tolerance to habitat conversion in tropical amphibians and reptiles

Habitat conversion is a major driver of the biodiversity crisis, yet why some species undergo local extinction while others thrive under novel conditions remains unclear. We suggest that focusing on species' niches, rather than traits, may provide the predictive power needed to forecast biodiversity change. We first examine two Neotropical frog congeners with drastically different affinities to deforestation and document how thermal niche explains deforestation tolerance. The more deforestation‐tolerant species is associated with warmer macroclimates across Costa Rica, and warmer microclimates within landscapes. Further, in laboratory experiments, the more deforestation‐tolerant species has critical thermal limits, and a jumping performance optimum, shifted ~2 °C warmer than those of the more forest‐affiliated species, corresponding to the ~3 °C difference in daytime maximum temperature that these species experience between habitats. Crucially, neither species strictly specializes on either habitat – instead habitat use is governed by regional environmental temperature. Both species track temperature along an elevational gradient, and shift their habitat use from cooler forest at lower elevations to warmer deforested pastures upslope. To generalize these conclusions, we expand our analysis to the entire mid‐elevational herpetological community of southern Costa Rica. We assess the climatological affinities of 33 amphibian and reptile species, showing that across both taxonomic classes, thermal niche predicts presence in deforested habitat as well as or better than many commonly used traits. These data suggest that warm‐adapted species carry a significant survival advantage amidst the synergistic impacts of land‐use conversion and climate change.     

Thermal death kinetics of fifth-instar Plodia interpunctella (Lepidoptera: Pyralidae)

Heat treatments have been suggested as alternatives to chemical fumigants for control of postharvest insects in dried fruits and nuts. Conventional forced hot air treatments heat product too slowly to be practical, but radio frequency treatments are capable of more rapid product heating. While developing radio frequency heat treatments for dried fruits and nuts, the heat tolerance of nondiapausing and diapausing fifth-instar larvae of the Indianmeal moth, Plodia interpunctella (Hübner), was determined using a heating block system developed by Washington State University. Both a 0.5th order kinetic model and a classical empirical model were used to estimate lethal exposure times for temperatures of 44-52 degrees C for nondiapausing fifth-instar larvae. We obtained 95% mortality at exposures suitable for practical radio frequency treatments (< or = 5 min) with temperatures of 50 and 52 degrees C. Diapausing larvae were significantly more tolerant than nondiapausing larvae at the lowest treatment temperature and shortest exposure, but differences were not significant at more extreme temperature-time combinations. Previous studies showed that fifth-instar larvae of the navel orangeworm, Amyelois transitella (Walker), were more heat tolerant than either diapausing or nondiapausing Indianmeal moth larvae. Consequently, efficacious treatments for navel orangeworm would also control Indianmeal moth.     

Viability and rate of development at different temperatures in Drosophila: a comparison of constant and alternating thermal regimes

Populations belonging to the sibling species Drosophila melanogaster and D. simulans were collected in Southwestern France and Southern Spain, and investigated under constant (CT) and alternating (AT) temperature regimes. Development under CT was possible between 11 and 32 degrees C and egg-to-adult viability curves were almost 'rectangular', with a sharp decrease below 14 and above 29 degrees C. Rate of development followed a complex non-linear curve. A model described the curve as an exponential below a critical temperature (T(C)), and above T(C) as the difference between this function and another exponential which is assumed to show deleterious effects of heat. Developmental rates under two daily 12-h phases with various mid-temperatures and thermal amplitudes were compared to expected rates calculated from the above model. Acceleration effects were observed at four AT (in increasing order: 12-30, 9-21, 11-21, 16-26 degrees C); retardation occurred at three other ones (in increasing order, 7-21, 5-15, 7-29 degrees C). When expressed by the ratio observed/expected, the effects could be predicted using a multiple regression, as a positive function of the thermal amplitude and a negative one of the mid-temperature. Viability under AT was analysed considering an equivalent developmental temperature (EDT), that is the CT which would produce the same rate or development. Very low viabilities occurred under broad amplitude regimes, but the deleterious effects of some extreme temperatures, that would be lethal under CT, could be recovered by daily return to a moderate temperature. The two species exhibited slight but significant differences in their characteristic temperatures: developmental zero, critical temperature, temperature of maximum rate, upper developmental limit. All data may be interpreted by considering that D. simulans compared to D. melanogaster is more tolerant to cold but less tolerant to heat.     

Resilience to extreme temperature events: acclimation capacity and body condition of a polymorphic fish in response to thermal stress

Considerable attention has been given to the potential impacts of global climate change on biodiversity. In the present study, we combine understudied themes by examining the ability of a freshwater fish (polymorphic for heat‐sensitivity) to respond to short‐term thermal stress mimicking an extreme temperature event. We simultaneously measured the effect of thermal stress on the body condition of heat‐sensitive and heat‐tolerant forms to evaluate an existing hypothesis regarding the underlying mechanism by which temperature affects the maintenance of genetic variation in this species. Surprisingly, the heat‐sensitive allelic variant increased in body condition equally as much as a heat‐tolerant variant under acute heat stress. More importantly, the heat‐sensitive variant exhibited a significant response to thermal stress, with an upward shift of greater than 2 °C in critical thermal maximum. Our findings suggest a complexity to the relationship between thermal stress and male body condition that may depend on an interaction with other factors such as resource level. Although the evolutionary fate of species with respect to climate change is typically evaluated in terms long‐term adaptive response, short‐term selection events could drastically reduce fitness and reduce evolutionary potential. Our results suggest that heat‐sensitive species may have considerably greater resilience to the short‐term, extreme perturbations to the environment that are expected under climate change. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 504–510.     

A heat wave and dispersal cause dominance shift and decrease biomass in experimental metacommunities

In experimental metacommunities with marine benthic microalgae, we tested whether heat stress changes effects of connectivity and habitat heterogeneity on metacommunity structure and functioning, by manipulating a simulated heat wave, dispersal frequency and a light intensity gradient. We found that all measures of mean local and regional diversity and community biomass significantly declined after the heat wave and showed no sign of recovery. Additionally, dispersal decreased diversity and increased dominance in both the heat stressed and control communities. Together the heat wave and high dispersal frequency induced a dominance shift by spreading a temperature tolerant but low yielding species from its source patches with low light intensity across the metacommunity, an effect that increased with time. Although different species became dominant at high dispersal frequency with and without the heat wave, the shift towards a temperature tolerant species was not sufficient to maintain total community biomass. Thus, short‐term disturbance may cause longer‐term loss of ecosystem function due to dominance shifts in the composition of communities. This study illustrates the importance of employing multispecies approaches when attempting to predict responses of communities to environmental changes.