Carbon emission associated with respiration and calcification of nine gastropod species from the intertidal rocky shore of Western Europe

Intertidal rocky shores are characterized by vertical zonation that results from the interplay between environmental conditions, organism physiology, and species interactions. Metabolism of intertidal organisms is highly variable between species and it changes with vertical position along the intertidal gradient. The present study aimed to quantify the carbon metabolism of nine intertidal rocky shore gastropods, in order to clarify their respective roles in carbon production during emersion and immersion. The influences of monthly temperature variation and tidal level were tested for each species. Analyses were performed in the laboratory using the infrared gas analyzer method for measuring aerial respiration rates, and the dissolved inorganic carbon and total alkalinity technique for measuring aquatic respiration rate and calcification. Hourly carbon fluxes were calculated for the mean annual temperature of 13 °C measured in both air and underwater in the study area. Respiration rates were similar for emersion (8–25 μmol CO₂ g AFDW^?1 h^?1) and immersion (10–23 μmol DIC g AFDW^?1 h^?1). For all species, underwater respiration fluxes were more influenced by monthly temperature variation than by air fluxes, probably as an adaptation to the rapid changes occurring during emersion. Calcification was an important factor influencing annual carbon fluxes for all studied species; every species showed different calcification rates according to its size and position on the intertidal zone. Annual carbon emissions were calculated using the mean immersion/emersion time of each species. Intertidal gastropod carbon emission was primarily influenced by body biomass and their vertical position within the intertidal zone.     

Ecological costs on local adaptation of an insect herbivore imposed by host plants and enemies

Herbivore populations may become adapted to the defenses of their local hosts, but the traits that maximize host exploitation may also carry ecological costs. We investigated the patterns and costs of local adaptation in the pine processionary moth, Thaumetopoea pityocampa, to its host plants, Pinus nigra and P. sylvestris. The two hosts differ in needle toughness, a major feeding impediment for leaf-eating insects. We observed a west-to-east gradient of increasing progeny size in the Italian Alps, matching the pattern in toughness of their respective local host plant. Eastern populations that feed on the native P. nigra with tough needles had larger eggs, and neonate larvae with larger head capsules, than western populations that feed on the native P. sylvestris and the introduced P. nigra with softer foliage. In a reciprocal transfer experiment that involved the eastern-most and the western-most populations of T. pityocampa from this region, and excluded natural enemies, we found evidence for local adaptation to the host plant. Specifically, larvae from the western population only performed well when raised on their local hosts with soft needles, and they suffered near-complete mortality on the tough foliage at the eastern site. In contrast, larvae from the eastern population survived equally well at both sites. Local adaptation involved a trade-off between progeny size and the number of offspring. We hypothesized that an additional cost, imposed by natural enemies, may be associated with increased egg size: we also observed a west-to-east gradient of increased egg parasitism. We tested this hypothesis in a common garden by exposing eggs of both populations to parasitism by two native egg parasitoids, Ooencyrtus pityocampae and Baryscapus servadeii. The eastern population suffered a higher level of parasitoid attack by O. pityocampae than the western population, and performance of hatched adults of both parasitoids was enhanced in large eggs. Thus, increased neonate quality (larger eggs yielding larger larvae) confers an advantage on tough foliage but incurs the ecological cost of increased parasitism, which may constrain further adaptation by this herbivore.     

Huddling up in a dry environment: the physiological benefits of aggregation in an intertidal gastropod

In many intertidal gastropods, the formation of aggregations and closing of the opercular opening are behaviors commonly assumed to be associated with water conservation and maintenance of body temperature during tidal emersion periods. In the laboratory, we quantified the relationship between these two behaviors in a littorinid snail common to the north-central shores of Chile, Echinolittorina peruviana, and evaluated any benefit of these behaviors during desiccating conditions. We predicted that solitary individuals would maintain their opercula open for less time than aggregated snails when exposed to drier conditions due, at least in part, to differences in evaporative water loss. In laboratory trials, where relative humidity was manipulated, we observed that aggregated snails maintained their opercula open for longer periods of time than solitary snails under increasingly drier conditions. These results, together with observations of body temperature, suggest that aggregated animals may able to maintain gaseous exchange with their environment for longer periods of time than solitary individuals in response to desiccation stress. Our results suggest an interactive effect of two behaviors that confer physiological benefits when confronted with extreme physical conditions experienced during periods of emersion.     

Abundance,population structure and claw morphology of the semi-terrestrial crab <Emphasis Type="Italic">Pachygrapsus marmoratus</Emphasis> (Fabricius, 1787) on shores of differing wave exposure

Wave action is known to influence the abundance and distribution of intertidal organisms. Wave action will also determine the duration and suitability of various foraging windows (high-tide and low-tide, day and night) for predation and can also affect predator behaviour, both directly by impeding prey handling and indirectly by influencing prey abundance. It remains uncertain whether semi-terrestrial mobile predators such as crabs which can access intertidal prey during emersion when the effects of wave action are minimal, are influenced by exposure. Here, we assessed the effect of wave action on the abundance and population structure (size and gender) of the semi-terrestrial intertidal crab Pachygrapsus marmoratus on rocky shores in Portugal. The activity of P. marmoratus with the tidal cycle on sheltered and exposed shores was established using baited pots at high-tide to examine whether there was activity during intertidal immersion and by low-tide searches. Because prey abundance varies along a wave exposure gradient on most Portuguese shores and because morphology of crab chelipeds are known to be related to diet composition, we further tested the hypothesis that predator stomach contents reflected differences in prey abundance along the horizontal gradient in wave exposure and that this would be correlated with the crab cheliped morphology. Thus, we examined phenotypic variation in P. marmoratus chelipeds across shores of differing exposure to wave action. P. marmoratus was only active during low-tide. Patterns of abundance and population structure of crabs did not vary with exposure to wave action. Stomach contents, however, varied significantly between shores of differing exposure with a higher consumption of hard-shelled prey (mussels) on exposed locations, where this type of prey is more abundant, and a higher consumption of barnacles on sheltered shores. Multivariate geometric analysis of crab claws showed that claws were significantly larger on exposed shores. There was a significant correlation between animals with larger claws and the abundance of mussels in their stomach. Variation in cheliped size may have resulted from differing food availability on sheltered and exposed shores.     

Genetic differentiation in the intertidal zone in populations of the alga Enteromorpha linza (Ulvales: Chlorophyta)

Genetic differentiation within the intertidal zone was examined in six populations of the asexually reproducing alga Enteromorpha linza growing in the Long Island Sound, USA. Four of the five populations sampled in 1981 showed significant differentiation between high and low intertidal positions with respect to the GOT-2 locus. The pattern of differentiation was consistent for samples collected at several times during the year with some seasonal modifications. Four additional polymorphic loci, resolved in 1982, identified a total of 13 five-locus genotypes or clones. Four of the six populations sampled in 1982 showed significant differences in clone frequency between high and low intertidal positions. Laboratory experiments revealed differences in response to temperature among the clones. At 24°C a high intertidal associated clone showed an increase in growth, while low intertidal associated clones showed decreased growth compared to growth at 15°C. These results suggest that the microgeographic differentiation observed for E. linza in the intertidal zone may in part be due to the differential adaptation of clones to different intertidal environments. Additional demographic information is needed for individual clones in order to determine the role longevity, reproductive output, recruitment and interclonal competition play in maintaining the observed differentiation.     

Climate vs. soil factors in local adaptation of two common plant species

Evolutionary theory suggests that divergent natural selection in heterogeneous environments can result in locally adapted plant genotypes. To understand local adaptation it is important to study the ecological factors responsible for divergent selection. At a continental scale, variation in climate can be important while at a local scale soil properties could also play a role. We designed an experiment aimed to disentangle the role of climate and (abiotic and biotic) soil properties in local adaptation of two common plant species. A grass (Holcus lanatus) and a legume (Lotus corniculatus), as well as their local soils, were reciprocally transplanted between three sites across an Atlantic-Continental gradient in Europe and grown in common gardens in either their home soil or foreign soils. Growth and reproductive traits were measured over two growing seasons. In both species, we found significant environmental and genetic effects on most of the growth and reproductive traits and a significant interaction between the two environmental effects of soil and climate. The grass species showed significant home site advantage in most of the fitness components, which indicated adaptation to climate. We found no indication that the grass was adapted to local soil conditions. The legume showed a significant home soil advantage for number of fruits only and thus a weak indication of adaptation to soil and no adaptation to climate. Our results show that the importance of climate and soil factors as drivers of local adaptation is species-dependent. This could be related to differences in interactions between plant species and soil biota.     

Effects of emersion and temperature upon growth of the tropical brown alga Padina japonica,using a tide-simulation apparatus

A simple tide-simulation apparatus was used to investigate the influence of emersion and temperature on the intertidal growth of Padina japonica Yamada in both the juvenile and adult growth phases. The upper zonation limits are not determined by the sensitivity of any particular growth phase, since all phases show the same emersion tolerance limits. The species grows best when continuously submerged, and growth rates decrease with increasing emersion up to a clear upper zonation boundary. High temperature reduces the emersion tolerance and is therefore an important factor in tropical intertidal zonation.     

Population structure and phylogeography of an acorn barnacle with induced defense and its gastropod predator in the Gulf of California

Using sequence variation in the mitochondrial cytochrome oxidase I locus, we estimated the population structure and phylogeography of the intertidal acorn barnacle, Chthamalus anisopoma, and its gastropod predator, Mexacanthina lugubris angelica. Both are endemic to the Gulf of California, being derived from taxa on the Pacific coast of the Baja peninsula, and both exhibit phenotypic plasticity for traits affecting their coevolutionary interactions. Consistent with expectations based on differences in dispersal, C. anisopoma populations generally lack geographic structure, while those of M. l. angelica are more strongly structured. However, the variable degree of differentiation in both species suggests that the extent of reciprocal selection and local adaptation in the species will vary geographically, a result consistent with the concept of a geographic mosaic of coevolution. The pattern of variation in C. anisopoma shows clear evidence of recent spatial expansion, possibly due to increased habitat availability following the last glacial maximum. Phylogeographic analyses suggest that M. l. angelica diversified into three distinct clades after the colonization of the Gulf. Overall, our results illustrate how dispersal potential, geological and climatic events, and recent population growth have impacted the pattern of sequence variation in the two species.     

Estimates of outcrossing rates and of inbreeding depression in a population of the marine angiosperm Zostera marina

Despite the great diversity of pollination and fertilization mechanisms observed in marine plants, little is known about the causes or maintenance of this variation. In this study, I estimated outcrossing rates and levels of inbreeding depression in Zostera marina L. (eelgrass), providing the first empirical test of hypotheses about the evolution of breeding systems in plants with submerged flowers. This study also addressed temporal separation of female and male flowering (dichogamy) in eelgrass as a mechanism promoting an outcrossing mating system, and whether the mating system in eelgrass is related to the degree of dichogamy in the field. Outcrossing rates (0<t<1) estimated from two polymorphic allozyme loci indicate that the Z. marina population in intertidal and subtidal habitats in False Bay, Washington, USA, was highly outcrossing in both 1991 (intertidal t=0.905, subtidal t=1.0) and 1992 (intertidal t=0.775, subtidal t=0.611). The outcrossing rates were positively associated with the degree of dichogamy in 1992; intertidal plants exhibited a greater temporal separation of female and male flowering and a higher outcrossing rate than did subtidal plants. Inbreeding depression at seed set was estimated from hand pollinations (self- and outcross) on 20 reproductive individuals from the False Bay population. Averaged across all maternal parents, a greater proportion of outcrossed flowers set seed than selfed flowers; i.e., inbreeding depression was detected. Plants exhibited genetic variation for inbreeding depression, detected as a significant pollination treatment × maternal family interaction in a log-likelihood analysis. By the end of the seed-maturation period (7 mo after intial seed set) some families showed outbreeding depression, i.e., greater fitness in progeny derived from selfing than in progeny from outcrossing. The inbreeding depression in the False Bay population may be an important selective factor contributing to the maintenance of dichogamy and an outcrossing mating system, as proposed for aquatic plants.     

High-temperature tolerance of photosynthesis in the red alga Chondrus crispus

Chondrus crispus (Stackhouse) is a perennial red seaweed, common in intertidal and shallow sublittoral communities throughout the North Atlantic Ocean. In the intertidal zone, C. crispus may experience rapid temperature changes of 10 to 20C° during a single immerison-emerision cycle, and may be exposed to temperatures that exceed the thermal limits for long-term survival. C. crispus collected year-round at Long Cove Point, Chamberlain, Maine, USA, during 1989 and 1990, underwent phenotypic acclimation to growth temperature in the laboratory. This phenotypic acclimation enhanced its ability to withstand brief exposure to extreme temperature. Plants grown at summer seawater temperature (20°C) were able to maintain constant rates of lightsaturated photosynthesis at 30°C for 9 h. In contrast, light-saturated photosynthetic rates of plants grown at winter seawater temperature (5°C) declined rapidly following exposure to 30°C, reached 20 to 25% of initial values within 10 min, and then remained constant at this level for 9 h. The degree of inhibition of photosynthesis at 30°C was also dependent upon light intensity. Inhibition was greatest in plants exposed to 30°C in darkness or high light (600 mol photons m^-2s^-1) than in plants maintained under moderate light levels (70 to 100 mol photons m^-2s^-1). Photosynthesis of 20°C-acclimated plants was inhibited by exposure to 30°C in darkness or high light, but the degree of inhibition was less than that exhibited by 5°C-grown plants. Not only was light-saturated photosynthesis of 20°C plants less severely inhibited by exposure to 30°C than that of 5°C plants, but the former also recovered faster when they were returned to growth conditions. The mechanistic basis of this acclimation to growth temperature is not clear. Our results indicate that there were no differences between 5 and 20°C-grown plants in the thermal stability of respiration, electron transport associated with Photosystems I or II, Rubisco or energy transfer between the phycobilisomes and Photosystem II. Overall, our results suggest that phenotypic acclimation to seawater temperature allows plants to tolerate higher temperatures, and may play an important role in the success of C. crispus in the intertidal environment.     

Nutritional status affects the capacity of the snail Concholepas concholepas to synthesize Hsp70 when exposed to stressors associated with tidal regimes in the intertidal zone

Synthesis of the heat shock protein Hsp70 is one of the most important physiological mechanisms that intertidal organisms possess to counteract damage to macromolecules caused by stressors associated with the tidal cycle. However, the synthesis and activity of Hsp70 involves an elevated energetic cost. We evaluated the effect of the nutritional status (fed vs. starved for 2 weeks) of juvenile Concholepas concholepas mollusc on their capacity to synthesize Hsp70 during emersion (i.e. low tide) and immersion (i.e. high tide) at high temperatures (24 °C, e.g. summer conditions) and at low temperatures (7 °C, e.g. winter conditions). In addition, we evaluated whether Hsp70 is induced directly upon exposure to stress (emersion) or during recovery (re-immersion). Starvation decreased the content of stored energy substrates of juveniles as well as their ability to synthesize Hsp70 during emersion under thermal stress, especially at high temperatures. Additionally, analysis of environmental factors associated with laboratory simulation of tidal regimes indicated that juveniles in starvation, in contrast to fed juveniles, did not significantly increase their levels of Hsp70 during cold emersion (7 °C) or warm emersion (24 °C) or upon re-immersion. Induction of Hsp70 occurred during exposure to stress (low-tide conditions) and not when juveniles returned to “normal” conditions (high-tide conditions). Thus, the synthesis of Hsp70 for the juveniles of this intertidal snail species was coordinated and adapted to the tidal cycle, and the species responds in a similar way to hot and cold emersion conditions. The observed levels of Hsp70 reflect the ability of the individual to synthesize these proteins, which is dependent on the nutritional status of the individual.     

Genetic differentiation between morphotypes in the Antarctic limpet <Emphasis Type="Italic">Nacella concinna</Emphasis> as revealed by inter-simple sequence repeat markers

The limpet Nacella concinna (Strebel 1908) was the focus of numerous studies dealing with Antarctic benthos. One of the main characteristics of the species is the presence of two distinguishable morphotypes, one inhabiting the intertidal (during summer) and the other inhabiting the subtidal. For a long time these forms were considered as an expression of phenotypic plasticity, since previous studies did not found genetic differences between them. In the present work, we performed both a morphometric and a genetic differentiation analysis (using ISSR-PCR markers) of these two forms in three stations sampled at Potter Cove, South Shetland Islands. The results confirmed the morphological differences between intertidal and subtidal forms reported in other Antarctic localities. The genetic differences detected indicate that the two forms can be considered as genetically distinct populations maintaining low levels of gene flow. The degree of reproductive isolation of the ecotypes is discussed, as well as the possible origin of the divergence. The genetic differentiation observed can also have behavioral and physiological correlates, pointing out the importance of taking into account the potential differences in the response of both populations to different conditions in future studies in this species. M. C. de Aranzamendi and R. Sahade contributed equally to this paper.     

No evidence for homeoviscous adaptation in intertidal snails: analysis of membrane fluidity during thermal acclimation, thermal acclimatization, and across thermal microhabitats

Many eurythermal organisms alter composition of their membranes to counter perturbing effects of environmental temperature variation on membrane fluidity, a process known as homeoviscous adaptation. Marine intertidal gastropods experience uniquely large thermal excursions that challenge the functional integrity of their membranes on tidal and seasonal timescales. This study measured and compared membrane fluidity in marine intertidal snail species under three scenarios: (1) laboratory thermal acclimation, (2) thermal acclimatization during a hot midday low tide, and (3) thermal acclimatization across the vertical intertidal zone gradient in temperature. For each scenario, we used fluorescence polarization of the membrane probe DPH to measure membrane fluidity in individual samples of gill and mantle tissue. A four-week thermal acclimation of Tegula funebralis to 5, 15, and 25°C did not induce differences in membrane fluidity. Littorina keenae sampled from two thermal microhabitats at the beginning and end of a hot midday low tide exhibited no significant differences in membrane fluidity, either as a function of time of day or as a function of thermal microhabitat, despite changes in body temperature up to 24°C within 8 h. Membrane fluidities of a diverse group of snails collected from high, middle, and low vertical regions of the intertidal zone varied among species but did not correlate with thermal microhabitat. Our data suggest intertidal gastropod snails do not exhibit homeoviscous adaptation of gill and mantle membranes. We discuss possible alternatives for how these organisms counter thermal excursions characteristic of the marine intertidal zone.     

Adaptations to semi-terrestrial life in embryos of East African mangrove crabs: a comparative approach

In this experimental study, we compared the embryonic respiration rate in air and water of six East African sesarmid species with intertidal, supratidal and arboreal habits, to highlight possible adaptations in embryonic metabolism to their different lifestyles. The embryos of all analysed crabs showed bimodal respiration, but we did not find a trend towards an enhanced embryonic oxygen uptake in air from the intertidal to the arboreal and supratidal species. However, the late-stage embryos of the most land-adapted species, Chiromantes spp., showed an enhanced metabolism when immersed in sea water that we interpreted as an adaptive recovery mechanism to cope with the storage of by-products due to marine-based metabolic pathways during long emersion periods. Thus, we showed that the embryos of land-adapted species, although still strongly water dependent, are well adapted to semi-terrestrial habitats and represent a minor limiting factor for females, which are not restricted in their emersion period by the oxygen requirements of their embryos.     

Latitude or biogeographic breaks? Determinants of phenotypic (co)variation in fitness-related traits in Betaeus truncatus along the Chilean coast

Ectothermal organisms distributed along environmental gradients in a wide geographical distribution display extensive phenotypic variation. This is particularly pervasive along latitudinal clines, which are linked to gradual changes in environmental factors. Widespread species may also be distributed among biogeographic breaks, which in contrast to smooth clines, often show abrupt changes in phenotypic traits. In species with widespread latitudinal distribution that also encompass important biogeographical breaks, it is not clear which of those factors prevails on shaping the phenotypic variation or if some traits are particularly more sensitive to one or the other. To evaluate this, we measured 4 fitness-related traits in 6 populations of the intertidal snapping shrimp Betaeus truncatus, as its distribution along Chile expands over 40° in latitude and three major biogeographical provinces. Here, we statistically evaluated the role of both, latitude and biogeographic breaks, on mean population values of fitness-related traits but also on the variances and covariances (i.e., P-matrix) between them. Overall, our results (1) indicate that latitude is more important than breaks in shaping the phenotypic variation of most of these fitness-related traits, (2) show that the differences in the variance–covariance relationship among traits between the extremes of the gradient arises from gradual increases in variance and rather sharp changes in covariance at mid-latitudes and (3) show that at present, it is difficult to unambiguously determine whether natural selection or plasticity is responsible for the observed pattern in means, variances and covariances and only further work might disentangle these possibilities.     

No evidence for genetic differentiation between Antarctic limpet Nacella concinna morphotypes

The extent to which genetic divergence can occur in the absence of physical barriers to gene flow is currently one of the most controversial topics in evolutionary biology, with implications for our understanding of speciation, phenotypic plasticity and adaptive potential. This is illustrated by a recent study reporting a surprising pattern of genetic differentiation between intertidal and subtidal morphotypes of the broadcast-spawning Antarctic limpet Nacella concinna. To explore this further, we collected almost 400 Antarctic limpets from four depths (intertidal, 6, 15 and 25 m) at Adelaide island, Antarctica, and conducted a combined morphometric and genetic analysis using 168 polymorphic amplified fragment length polymorphism (AFLP) loci. Morphological analysis revealed not only pronounced differences between the two morphotypes, but also a continuous cline in shell shape from the intertidal zone down to 25 m depth, suggesting that the distinction between the morphotypes may be artificial. Moreover, genetic analysis using both F _st and a Bayesian analogue found no evidence for differentiation either between the two morphotypes or by depth, and a Bayesian cluster analysis did not detect any cryptic genetic structure. Our findings lend support to the notion that limpets can be phenotypically highly plastic, although further studies are required to determine unequivocally whether there is any genetic basis to the observed variation in shell morphology.     

Reproductive isolation between intertidal and subtidal<Emphasis Type="Italic"> Scoloplos armiger</Emphasis> (Polychaeta,Orbiniidae) indicates sibling species in the North Sea

Two distinct modes of development in the common polychaete Scoloplos armiger (O. F. Müller, 1776) occur in the North Sea region: holobenthic development in egg cocoons and pelagic larvae hatching from suspended eggs. In the northern Wadden Sea near the island of Sylt, we observed that egg cocoons are produced intertidally while pelagic larvae originate from the adjacent subtidal zone. A previous genetic comparison between these subtidal and intertidal populations revealed distinct gene pools, suggesting that reproductive differences are not phenotypic but heritable. In this study, crossbreeding experiments show that intertidal and subtidal populations are reproductively isolated. Couples with males and females from different habitats had no offspring. Production of egg cocoons is determined by female origin from the intertidal zone. Pelagic larvae occurred only in couples with subtidal females and subtidal males. Intertidal males have spermatozoa with heads twice as long as those from subtidal males and a significantly shorter flagellum. We suspect that deviating sperm morphology may cause the reproductive breakdown at the fertilization stage. Juveniles hatching from cocoons have shorter anal cirri compared to juveniles that metamorphosed from pelagic larvae. We conclude there to be two sympatric sibling species in S. armiger: 'type I' in intertidal areas, which have egg cocoons, no pelagic larvae, elongated sperm heads, shortened sperm flagella and anal cirri; and a subtidal 'type S', lacking egg cocoons but with pelagic larvae, short sperm heads, long sperm flagella and anal cirri.Communicated by O. Kinne, Oldendorf/Luhe     

Geographic variation,niche breadth and genetic differentiation at different geographic scales in the mussels Mytilus californianus and M. edulis

Genetic differentiation was investigated in the marine mussels Mytilus californianus Conrad and M. edulis Linn. from the west coast of North America. In allopatry with M. californianus, M. edulis occurs throughout the intertidal zone; however, in microgeographic sympatry its ecological range is restricted to above the M. californianus mussel bed and to patches of substratum opened by natural disturbances within the bed. Over the same geographic scale, the broader-niched M. edulis shows greater among-locality genetic difference and greater levels of polymorphism than M. californianus at two enzyme loci. Genetic differentiation on a geographic basis was investigated in M. californianus at a single rock (on a scale of meters), on an island (on a scale of kilometers), throughout a strait (on a scale of 10² kilometers), and along the west coast of North America (on a scale of 10³ km). Differentiation was minimal over the west coast, and could be explained by microhabitat differences in a local area. The minimal differentiation in west coast mussels relative to strong geographic differentiation of M. edulis on the east coast of North America may be related to the steeper latitudinal thermal gradient on the east coast. Local ecologically related microgeographic variation can result in biased and misleading estimates of genetic heterogeneity but microgeographic variation at enzyme loci may not be due to selection at the loci investigated or may even be due to the breeding structure of the mussel populations, as suggested by Tracey et al. (1975).