Spatial and seasonal variation in δ15N and δ13C values in a mesopredator shark, Squalus suckleyi, revealed through multitissue analyses

We used variance decomposition to explore the importance of body size, sex, location, and sampling period as predictors of intrapopulation variation in δ¹⁵N and δ¹³C values in spiny dogfish Squalus suckleyi from the Puget Sound–Strait of Georgia basin. Isotopes in two tissues with long (dorsal white muscle) and short (liver) isotopic turnover rates (~1 year and ~3–4 months, respectively) were sampled to evaluate whether the relative importance of each variable differed depending on the time span over which diet information was integrated. Significant spatial variation was observed in both muscle and liver isotopic composition, whereby location uniquely explained 25 and 17 % of the total variance, respectively. The remaining variables explained considerably less variation in both tissue types. Furthermore, evidence of seasonal isotopic shifts in δ¹⁵N and δ¹³C values was apparent, but differed widely in direction and magnitude among groups. These findings suggest that members of spiny dogfish schools may share a common feeding history, possibly by spending extended time periods (weeks to months) foraging in a spatially fixed region. Another explanation is that individuals may move and feed in aggregations that exist for extended periods. These complex group-level patterns suggest that even for large-bodied, motile predators such as sharks, population-level diet estimates derived from averaging isotope ratios of individuals collected from only a few locations may poorly reflect the true population mean.     

High turnover of inorganic carbon in kelp habitats as a cause of δ13C variability in marine food webs

In a study to assess qualitatively the importance of organic matter derived from kelp production in the Aleutian Islands of subarctic Alaka, replicated samples of autotrophic sources and primary and secondary consumer organisms were sampled for ¹³C among sources, sites, (treatment) islands, and years. Unanticipated variation in the ¹³C of kelps occurred among overtly similar sites at different islands. Variation in the ¹³C of the surface canopy-forming kelp Alaria fistulosa was particularly extreme, ranging from-15.5 to-28.0 compared to the understory kelps, Laminaria spp. A. fistulosa ¹³C varied by as much as 6 to 7 among similar sites at a given island within years, and by as much as 3 to 4 between years at the same sampling site. In serveral cases, ¹³C variation was weakly tracked by some consumer organisms, suggesting that even detritus pathways through the food web can be localized and tightly coupled. Dynamic cycles in the concentration and ¹³C of dissolved inorganic carbon (DIC) and aqueous CO₂ concentration ([CO₂]_aq) were measured at three sites on one island. The ¹³C or organic carbon fixed by A. fistulosa, calculated from diurnal DIC concentration and ¹³C measurements, varied by 15 and varied inversely with [CO₂]_aq concentrations. Local DIC variability, probably resulting from high productivity and decreased turbulence in dense kelp habitats, provides a possible mechanism of variation in kelp ¹³C. The short-term variability in the ¹³C of organic carbon fixed by kelps indicates that sampling methodology and design must assess this potential variation in marine macrophyte ¹³C before making assumptions about the transfer of ¹³C-invariate organic matter to higher trophic levels. On the positive side, a predictable relationship between [CO₂]_aq concentration and kelp ¹³C offers a potentially robust means to assess productivity effects on CO₂ limination in kelps and other complex aquatic macrophyte habitats.     

δ15N and δ13C values in dental collagen as a proxy for age- and sex-related variation in foraging strategies of California sea lions

We assessed the foraging habits of California sea lions, Zalophus californianus, from Isla Santa Margarita, BCS, Mexico, by analyzing δ¹³C and δ¹⁵N values of dentin collagen. Since dentin is deposited annually in growth layer groups (GLGs), it can be subsampled to construct ontogenetic isotopic profiles at the individual level. We drilled 20 canine teeth and obtained 141 samples for isotopic analysis that were assigned to age-specific categories from GLG-based estimated ages. Pups’ GLGs had the highest mean δ¹⁵N values and the lowest mean δ¹³C values, a pattern likely driven by the consumption of milk. Juveniles had δ¹⁵N values between those of pups and adult females, which may reflect continued nursing into the second year or preferential consumption of coastal benthic versus pelagic prey. Significant differences were observed between the sexes of adults; adult females had lower mean δ¹³C and δ¹⁵N values than adult males. Higher isotope values in adult males relative to females may reflect a higher trophic position, but differences in foraging grounds cannot be excluded as a potential explanation because tracking data are not available at this time. Evidence of intra-specific foraging diversification may be related to a strategy to reduce competition within and among age and sex categories.     

Species-specific variation in cuttlebone δ13C and δ18O for three species of Mediterranean cuttlefish

Stable carbon (δ¹³C) and oxygen (δ¹⁸O) isotopes in cuttlebones of three species of Mediterranean cuttlefish (Sepia elegans, S. officinalis, and S. orbignyana) with different life histories were contrasted. Cuttlebone δ¹³C and δ¹⁸O were quantified at both the core and edge (representing early life and recent deposition, respectively) for all three species sampled from the southern Adriatic Sea in 2010 (n = 28). For S. officinalis, cuttlebone δ¹³C and δ¹⁸O values were both lower relative to S. elegans and S. orbignyana at the core by approximately 1.0–2.0 and 3.0 ‰, respectively. Differences between core and edge in cuttlebone δ¹³C and δ¹⁸O were also observed for S. officinalis with observed values at the cuttlebone edge (recent) exceeding core (early life) values by 2.5 ‰ for δ¹³C and 1.4 ‰ for δ¹⁸O. Differences in isotopic composition across S. officinalis cuttlebones are possibly reflective of ontogenetic migrations from nearshore nurseries (lower seawater δ¹³C and δ¹⁸O values) to offshore overwintering habitats (higher seawater δ¹³C and δ¹⁸O values). Overall, results from this study suggest that cuttlebone δ¹³C and δ¹⁸O hold promise as natural tags for determining the degree of spatial connectivity between nearshore and offshore environments used by cuttlefish.     

Differences in δ13C and δ15N values between feathers and blood of seabird chicks: implications for non-invasive isotopic investigations

Blood and feathers are the most targeted tissues for isotopic investigations in avian ecology, primarily because they can be easily and non-destructively sampled on live individuals. Comparing blood and feather isotopic ratios can provide valuable information on dietary shifts, trophic specialization and migration patterns, but it requires a good knowledge of the isotopic differences between the two tissues. Here, δ¹³C and δ¹⁵N values of whole blood (in blood cells of a few species) and simultaneously grown body feathers were measured in seabird chicks to quantify the tissue-related isotopic differences. Seabirds include 27 populations of 22 wild species that were sampled in 2000–2008, and a review of the literature added 8 groups (including adult birds) to the analysis. The use of a large data set that overall encompasses wide δ¹³C and δ¹⁵N ranges allowed us to depict for the first time accurate relationships between blood and feather isotopic ratios across avian taxa. Blood was impoverished in ¹³C and generally in ¹⁵N compared with feathers. Both mean δ¹³C and δ¹⁵N values of feathers and blood were highly positively and linearly related [feather δ¹³C = 0.972 (±0.020) blood δ¹³C + 0.962 (±0.414), and feather δ¹⁵N = 1.014 (±0.056) blood + 0.447 (±0.665), respectively; both P < 0.0001]. The regressions should be applied to mathematically correct feather or whole blood δ¹³C and δ¹⁵N values when comparing isotopic ratios within and between ecological studies on birds.     

Temporal and spatial variation in the δ<Superscript>15</Superscript>N and δ<Superscript>13</Superscript>C values of fish and squid from Alaskan waters

To test the hypothesis that stable isotope ratios from marine organisms vary, the δ¹⁵N and δ¹³C values from fish and squid collected in Alaskan waters were measured across years (1997, 2000, and 2005), seasons, geographic locations, and different size/age classes, and between muscle tissue and whole animals. Temporal, geographic, and ontogenetic differences in stable isotope ratios ranged from 0.5–2.5‰ (δ¹⁵N) to 0.5–2.0‰ (δ¹³C). Twenty-one comparisons of stable isotope values between whole organisms and muscle tissue revealed only four small differences each for δ¹⁵N and δ¹³C, making costly and space prohibitive collection of whole animals unnecessary. The data from this study indicate that significant variations of stable isotope values from animals in marine systems necessitates collection of prey and predator tissues from the same time and place for best interpretation of stable isotope analysis in foraging ecology studies.     

Carbon dioxide use by chemoautotrophic endosymbionts of hydrothermal vent vestimentiferans: affinities for carbon dioxide, absence of carboxysomes, and δ13C values

The hydrothermal vent vestimentiferans Riftia pachyptila Jones, 1981 and Ridgeia piscesae Jones, 1985 live in habitats with different abundances of external CO₂. R. pachyptila is found in areas with a high input of hydrothermal fluid, and therefore with a high [CO₂]. R. piscesae is found in a range of habitats with low to high levels of hydrothermal fluid input, with a correspondingly broad range of CO₂ concentrations. We examined the strategies for dissolved inorganic carbon (DIC) use by the symbionts from these two species. R. pachyptila were collected from the East Pacific Rise (9°50′N; 104°20′W) in March 1996, and R. piscesae were collected from the Juan de Fuca Ridge (47°57′N; 129°07′W) during September of 1996 and 1997. The differences in the hosts' habitats were reflected by the internal pools of DIC in these organisms. The concentrations of DIC in coelomic fluid from R. piscesae were 3.1 to 10.5 mM, lower than those previously reported for R. pachyptila, which often exceed 30 mM. When symbionts from both hosts were incubated at in situ pressures, their carbon fixation rates increased with the extracellular concentration of CO₂, and not HCO₃ ⁻, and symbionts from R. piscesae had a higher affinity for CO₂ than those from R. pachyptila (K _1/2 of 7.6 μM versus 49 μM). Transmission electron micrographs showed that symbionts from R. piscesae lack carboxysomes, irrespective of the coelomic fluid [DIC] of their host. This suggests that the higher affinity for CO₂ of R. piscesae symbionts may be their sole means of compensating for lower DIC concentrations. The δ¹³C values of tissues from R. piscesae with higher [DIC] in the coelomic fluid were more positive, opposite to the trend previously described for other autotrophs. Factors which may contribute to this trend are discussed. Received: 24 September 1998 / Accepted: 12 May 1999     

Determination of δ13C and δ15N and trophic fractionation in jellyfish: implications for food web ecology

Application of stable isotope analysis (SIA) in jellyfish allows definition of trophic patterns not detectable using gut content analysis alone, but analytical protocols require standardization to avoid bias in interpreting isotopic data. We determined δ¹³C and δ¹⁵N in Aurelia sp. from the northern Gulf of Mexico (30°00′N, 89°00′W–30°24′N, 88°00′W) to define differences in stable isotope composition between body parts and whole body, the effect of lipid extraction on δ¹³C in tissues, and fractionation values from medusa to prey. The isotopic composition of bell and whole Aurelia sp. was not different. The increase in δ¹³C values after lipid removal suggested a correction is needed. To aid future analyses, we derived a correction equation from empirical data for jellyfish samples. Laboratory feeding experiments indicated medusae increased +4 ‰ in δ¹³C and +0.1 ‰ in δ¹⁵N compared to their diet. These results suggest protocols commonly applied for other species may be inaccurate to define Aurelia sp. trophic ecology. Because Aurelia spp. are commonly found in marine ecosystems, accurately defining their trophic role by use of SIA has implications for understanding marine food webs worldwide.     

Detrital flows through the feeding pathway of the oyster (Crassostrea gigas) in a tropical shallow lagoon: δ13C signals

The spatial relationships and linkage of the detrital flows among the water column, the sediment and the oyster Crassostrea gigas cultured in the water column were examined by using stable carbon isotopes (δ¹³C) in a tropical shallow lagoon from October 1996 to June 1997. The lagoon is located in southwestern Taiwan and is isolated from the sea by sand barriers except at two tidal inlets. It receives freshwater mainly from two rivers. A total of 12 stations were set up along three transect lines, each running across the lagoon from riverine to tidal inlet localities. The δ¹³C values of the water-column POM exhibited a marked sea–river gradient, with values depleted from a high of −21.7‰ at seaward stations to a low of −28.2‰ at riverine stations; those in the sedimentary POM (<62 μm grain size) also revealed this trend, but to a lesser extent. Oysters of two known ages, 6 months old (“old oysters”) and newly settled individuals (“young oysters”), were transplanted from one station to each of the remaining stations, while some were left at the original station. Values of δ¹³C in the muscle of transplanted oysters changed in parallel with the sea–river gradient of δ¹³C in POM (decreasing from −16.0 to −18.5‰ in old oysters and from −16.8 to −21.9‰ in young ones). The spatial sea–river gradient of the oyster's δ¹³C is related not only to the distance between the site that the oyster inhabits and sea or riverine environment, but also to the tidal flow pattern that surrounds its feeding place. Although the δ¹³C value of the sedimentary POM was correlated with that of the water-column POM, the δ¹³C value of the oyster tissue was significantly correlated with that of the water-column POM, but not with that of the sedimentary POM. This suggests that the oyster feeds primarily on water-column rather than sedimentary POM. Received: 30 April 1999 / Accepted: 15 December 1999     

Differences in diet composition and foraging patterns between sexes of the Magellanic penguin (Spheniscus magellanicus) during the non-breeding period as revealed by δ13C and δ15N values in feathers and bone

In diving seabirds, sexual dimorphism in size often results in sex-related differences of foraging patterns. Previous research on Magellanic penguins, conducted during the breeding season, failed to reveal consistent differences between the sexes on foraging behavior, despite sexual dimorphism. In this paper, we tested the hypothesis that male and female Magellanic penguins differ in diet and foraging patterns during the non-breeding period when the constraints imposed by chick rearing activities vanish. We used stable isotope ratios of carbon and nitrogen in feather and bone to characterize the diet and foraging patterns of male and female penguins in the South Atlantic at the beginning of the 2009–2010 and 2010–2011 post-breeding seasons (feathers) and over several consecutive breeding and migratory seasons (bone). The mean δ¹³C and δ¹⁵N values of feathers showed no differences between the sexes in any of the three regions considered or in the diet composition between the sexes from identical breeding regions; however, Bayesian ellipses showed a higher isotopic niche width in males at the beginning of the post-breeding season. Stable isotope ratios in bone revealed the enrichment of males with δ¹³C compared with females across the three regions considered. Furthermore, the Bayesian ellipses were larger for males and encompassed those of females in two of the three regions analyzed. These results suggest a differential use of winter resources between the sexes, with males typically showing a larger diversity of foraging/migratory strategies. The results also show that dietary differences between male and female Magellanic penguins may occur once the constraints imposed by chick rearing activities cease at the beginning of the post-breeding season.     

Allometry of gill weights,gill surface areas,and foot biomass δ<Superscript>13</Superscript>C values of the chemoautotroph–bivalve symbiosis<Emphasis Type="Italic"> Solemya velum</Emphasis>

The protobranch bivalve Solemya velum Say, 1822 has large gills, which harbor chemolithoautotrophic bacteria that supply the majority of the clams organic carbon. A substantial portion of the CO₂, O₂, H₂S, and other nutrients necessary for symbiont autotrophy and host heterotrophy are acquired from the environment through the gills, whose large size may be necessary to facilitate the acquisition of sufficient O₂ from S. velums habitat to meet the combined demands of the host and symbionts. Large gills may also result in an oversupply of CO₂, which may in turn be responsible for the isotopically depleted ¹³C values observed in S. velum biomass (–31 to –34). Alternatively, gill hypertrophy may simply be an adaptation to house a large population of symbionts adjacent to their environmental source of dissolved gases and other nutrients. To better understand gill function in this symbiosis, gill weights, gill surface areas, and foot ¹³C values were measured as a function of total weights. S. velum gill weights were found to be a substantial portion of total clam weight, averaging 38% of wet weight, compared to nonsymbiotic protobranch bivalves Yoldia limatula Say, 1831 (5%) and Nucula proxima Say, 1822 (11%). Gill weights are a smaller percentage of total weight in larger individuals; the allometric equation for gill weight (G) as a function of total weight (M) is G=0.26M^0.85. Dry weights scale similarly. Gill surface areas are immense; the average gill surface area measured was 107 cm² g^–1 total soft tissue wet weight, the highest value for any marine invertebrate. Gill surface area (SA) also scales with size (SA=69.8M^0.85). When gill surface areas were calculated with respect to gill wet weights, they did not scale with size. The ¹³C values do not scale with size either, consistent with high rates of CO₂ supply at all sizes. Extraordinarily high rates of CO₂ supply relative to demand are supported by a model for CO₂ delivery based on Ficks law and the allometric relationship between surface areas and total weight, consistent with a role for large gill surface areas in the generation of isotopically depleted tissue ¹³C values.Communicated by J. P. Grassle, New Brunswick     

Variations in δ13C of water–soluble leaf and phloem organic matter of Platycladus orientalis: influences of photosynthetic and post–photosynthetic fractionation

Analysis of variations in water–soluble organic matter (WSOM) δ¹³C of leaves and phloem can efficiently describe the δ¹³C distributions within plants and identify the temporal variation of δ¹³C. In this study, WSOM δ¹³C values of both leaves and phloem (twig, stem, and root) of Platycladus orientalis were measured during seven sunny days, including 2–hour interval measurements at three days for diel pattern analysis and 6–hour interval measurements at the remaining four days for day–to–day variation analysis. Analysis of WSOM δ¹³C in different plant organs showed that ¹³C was generally depleted from leaves to twigs, then enriched in stems and subsequently depleted in roots. Stems were significantly ¹³C–enriched compared to twigs (p?<?0.05), while δ¹³C differences between stems and other organs and among leaves, twigs and roots were not significant (p?>?0.05). No clear diel patterns in δ¹³C of leaves and phloem were found. Daily average δ¹³C values indicated that all plant organs had more positive values on sunny days during the dry season than during the wet season. Both photosynthetic and post–photosynthetic fractionation influence variations in WSOM δ¹³C. These results have implications for research on plant physiology and plant water use.     

Accumulation, contamination, and seasonal variability of trace metals in the coastal zone – patterns in a seagrass meadow from the Mediterranean

Amongst a plethora of threats to seagrass ecosystems, contamination with heavy metals may well be one of the most significant. We therefore set out to track contamination levels with Cu, Zn, Pb and Cd in the principal autotrophic compartments and sediments of a meadow of Posidonia oceanica in the Gulf of Naples, Mediterranean Sea. With respect to metal levels, leaves and their associated epibiota are certainly not a homogenous compartment, as might perhaps be inferred from the common use of the term “leaf–epiphyte complex” in the literature. Save for Cu, all metal species analysed showed appreciable differences in concentration between seagrass leaves and epibiota. These results give strength to our argument that in ecotoxicological work leaves and epibiota should not be treated as a single unit. Although absolute differences in trace-metal levels among sampling periods varied somewhat with the specific component analysed (i.e. macrophyte organs, epibiota, sediment), an overall trend of markedly higher heavy-metal levels during the winter season is a striking one. Whilst annual cycles in growth dynamics of the seagrasses explain a significant fraction of the temporal variance, seasonality in productivity is a doubtful explanation for similar patterns in non-living sedimentary components; consideration of additional variables therefore seems sensible. As variables with consistent explanatory powers we suggest: (1) seasonal cycles in storm frequency and amplitude which remobilise metals bound in the sediments of the sea floor, and (2) increased precipitation during the cold season which may significantly increase marine metal levels through elevated weathering of rocks and elevated fluvial inputs of anthropogenic contaminant loads. Whereas Cd and Pb concentrations in seagrass leaves from the Gulf of Naples fall within the range for coastal areas subjected to low levels of heavy-metal pollution, Cu and Zn reach levels typical of highly contaminated regions, such as the waters bordering major coastal cities. Any direct comparisons of the pollution status of seagrass beds between different geographic areas are, however, likely to be confounded by the indiscriminate application of the “leaf–epiphyte complex”: the magnitude of the confounding effect depends on the ratio of epibiota/leave biomass, time of sampling, and metal species analysed. Received: 15 May 1997 / Accepted: 2 February 1998     

Matching and mismatching stable isotope (δ13C and δ15N) ratios in fin and muscle tissue among fish species: a critical review

Using non-lethal tissue sampling for stable isotope analysis has become standard in many fields, but not for fishes, despite being desirable when species are rare or protected, when repeated sampling of individuals is required or where removal may bias other analyses. Here, we examine the utility of fish dorsal fin membrane as an alternative to muscle for analyzing δ¹³C and δ¹⁵N ratios in two reef fish species (blue cod Parapercis colias and spotty Notolabrus celidotus) that have differing feeding modes. Both species exhibited evidence of size-based feeding from fin δ¹⁵N values, but not from muscle. Blue cod fin δ¹⁵N increased steadily throughout the sampled size range (213–412 mm fork length), whereas spotty exhibited a distinct ontogenetic diet shift at approximately 120–140 mm fork length after which size-based feeding did not occur. Fin membrane was higher than muscle in δ¹³C in both species and in δ¹⁵N for blue cod, but fin δ¹⁵N was lower than muscle in spotty. The δ¹³C and δ¹⁵N fin–muscle offsets were constant in spotty regardless of size, while in blue cod, δ¹³C was constant with fish size, but δ¹⁵N offsets increased with increasing fish size. Non-lethal sampling utilizing fin tissue can be employed to estimate stable isotope values of muscle in fishes, but it is necessary to assess relationships among tissues and the effects of fish size on isotope values a priori for each species studied. Our data indicated that fin membrane may be a more sensitive tissue than muscle for detecting size-based feeding in some fish species using stable isotopes. A critical literature review revealed inconsistencies in tissue types tested, little understanding of tissue-specific trophic shift or turnover rates, and pseudo-replicated analyses leading to erroneous postulating of 1:1 relationships between tissues.     

The metabolism and distribution of [14C‐phenyl]‐ethyl parathion in a tropical soil under field conditions

The persistence, distribution and metabolism of [^l4C‐phenyl]‐ethyl parathion applied to soil columns in an agricultural farm were investigated under tropical field conditions. Volatilization, soil microbial activity, moisture levels and pH were found to influence the persistence, distribution and metabolism of this pesticide in the soil. There was rapid distribution of the pesticide in the soil matrix with time resulting in high levels of bound residues which reduced the overall rate of disappearance of parathion residues from the soil. The soil was slightly acidic but the metabolites, paraoxon, p‐nitrophenol and p‐aminophenol were detected in the soil extracts 7 days after pesticide application. After 72 days, 43.7% of the applied pesticide remained in soil composed of 18.9% extractable and 24.8% bound residues. A mechanism for the metabolism of ethyl parathion in this soil is given.     

Longitudinal and vertical trends in stable isotope signatures (δ13C and δ15N) of omnivorous and carnivorous copepods across the South Atlantic Ocean

Stable isotope (SI) ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) were measured in omnivorous and carnivorous deep-sea copepods of the families Euchaetidae and Aetideidae across the Atlantic sector of the Southern Ocean to establish their trophic positions. Due to high and variable C/N ratios related to differences in lipid content, δ¹³C was corrected using a lipid-normalisation model. δ¹⁵N signals ranged from 3.0–6.9‰ in mesopelagic species to 7.0–9.5‰ in bathypelagic congeners. Among the carnivorous Paraeuchaeta species, the epi- to mesopelagic species Paraeuchaeta antarctica had lower δ¹⁵N values than the mesopelagic P. rasa and bathypelagic P. barbata. The same trend was observed among omnivorous Aetideidae, but was not significant. In the most abundant species P. antarctica, individuals from the western Atlantic had higher δ¹³C and δ¹⁵N values than specimens at the eastern stations. These longitudinal changes in δ¹³C and δ¹⁵N values were attributed to regional differences in hydrography and sea surface temperature (SST), in particular related to a northward extension of the Antarctic Polar Front (APF) at the easternmost stations. The results indicate that even in a mesopelagic carnivorous species, the changes in surface stable isotope signatures are pronounced.     

Stable isotopes (δ13C, δ15N) and modelling as tools to estimate the trophic ecology of cultivated oysters in two contrasting environments

Food sources for cultivated marine bivalves generally are not well identified, although they are essential for a better understanding of coastal ecosystems and for the sustainability of shellfish farming activities. In addition to phytoplankton, other organic matter sources (OMS), such as microphytobenthos and detritus (of terrestrial or marine origins), can contribute significantly to the growth of marine bivalves. The aim of this study was to identify the potential food sources and to estimate their contributions to the growth of the Pacific oyster (Crassostrea gigas) in two contrasting trophic environments of Normandy (France): the Baie des Veys (BDV) and the Lingreville area (LIN). Two sites were studied in the BDV area (BDV-S and BDV-N) and one in the LIN area. To estimate the contribution of each type of OMS, we used a combination of stable natural isotope composition (δ¹³C, δ¹⁵N) analysis of OMS and oyster tissue together with a modelling exercise. Field sampling was conducted every 2 months over 1 year. The sampled sources were suspended particulate organic matter from marine (PhyOM) and terrestrial (TOM) origins, microphytobenthos (MPB), detrital organic matter from the superficial sediment (SOM), and macroalgae (Ulva sp., ULV). A statistical mixing model coupled to a bioenergetic model was used to calculate the contributions of each different source at different seasons. Results showed that isotopic composition of the animal flesh varied with respect to the potential OMS over the year within each ecosystem. Significant differences were also observed among the three locations. For instance, the δ¹³C and δ¹⁵N values of the oysters ranged from −20.0 to −19.1‰ and from 6.9 to 10.8‰ at BDV-S, from −19.4 to −18.1‰ and from 6.4 to 10.0‰ at BDV-N, and from −21.8 to −19.4‰ and from 6.3 to 8.3‰ at LIN. The contributions of the different sources to oyster growth differed depending on the ecosystem and on the period of the year. Phytoplankton (PhyOM) predominated as the principal food source for oysters (particularly in the LIN location). MPB, TOM, and ULV detritus also possibly contributed to oysters’ diet during summer and autumn at the BDV-S and BDV-N sites. SOM was not considered an OMS because it was already a mix of the other four OMS, but rather a trophic reservoir that potentially mirrored the trophic functioning of marine ecosystems.     

Tissue,ontogenic and sex-related differences in δ<Superscript>13</Superscript>C and δ<Superscript>15</Superscript>N values of the oceanic squid <Emphasis Type="Italic">Todarodes filippovae</Emphasis> (Cephalopoda: Ommastrephidae)

Stable isotopes are increasingly used in the study of trophic interactions of many aquatic animals and most recently cephalopods. To evaluate the application of the method to squids, it is important to assess isotopic differences among and within consumer tissues that may confound the resolution of ecological relationships. Inter- and intra-tissue isotopic variation was examined in 55 individuals of the oceanic squid Todarodes filippovae that were collected at the beginning of April 2000 in the southwestern Indian Ocean (between 44°S, 76°E, and Saint Paul and Amsterdam islands, 38°S, 78°E). Delipidated soft tissues (mantle, arm, buccal mass, gill and reproductive organs) showed small δ¹³C and δ¹⁵N differences, which were probably tissue-specific. A lower carbon value was observed in the digestive gland as a consequence of incomplete lipid removal. Hard tissues, such as beaks and gladii, had lower ¹⁵N values than soft tissues, which can be explained by the presence of chitin, a ¹⁵N-depleted molecule. Females (n = 38) and males (n = 17) had identical δ¹³C values, but females showed higher δ¹⁵N values than males. The difference was size-related rather than sex-related, however, as females were generally larger than males. A comparison of similar-sized females and males produced identical nitrogen values. These data suggest dietary shifts from lower to higher trophic levels during growth, because δ¹⁵N values of large T. filippovae were much higher than that of small specimens. As expected, nitrogen values of lower beaks and gladii of large squids increased from the oldest to the most recently formed region, reflecting the progressive growth of chitinized tissues in parallel with dietary changes. Sequential sampling along the growth increments of squid beaks and gladii can likely be used to produce a chronological record of dietary information throughout an individual’s history.     

The synergistic effects of increasing temperature and CO2 levels on activity capacity and acid–base balance in the spider crab, Hyas araneus

With global climate change, ocean warming and acidification occur concomitantly. In this study, we tested the hypothesis that increasing CO₂ levels affect the acid–base balance and reduce the activity capacity of the Arctic spider crab Hyas araneus, especially at the limits of thermal tolerance. Crabs were acclimated to projected oceanic CO₂ levels for 12 days (today: 380, towards the year 2100: 750 and 1,120 and beyond: 3,000 μatm) and at two temperatures (1 and 4 °C). Effects of these treatments on the righting response (RR) were determined (1) at acclimation temperatures followed by (2) righting when exposed to an additional acute (15 min) heat stress at 12 °C. Prior to (resting) and after the consecutive stresses of combined righting activity and heat exposure, acid–base status and lactate contents were measured in the haemolymph. Under resting conditions, CO₂ caused a decrease in haemolymph pH and an increase in oxygen partial pressure. Despite some buffering via an accumulation of bicarbonate, the extracellular acidosis remained uncompensated at 1 °C, a trend exacerbated when animals were acclimated to 4 °C. The additional combined exposure to activity and heat had only a slight effect on blood gas and acid–base status. Righting activity in all crabs incubated at 1 and 4 °C was unaffected by elevated CO₂ levels or acute heat stress but was significantly reduced when both stressors acted synergistically. This impact was much stronger in the group acclimated at 1 °C where some individuals acclimated to high CO₂ levels stopped responding. Lactate only accumulated in the haemolymph after combined righting and heat stress. In the group acclimated to 1 °C, lactate content was highest under normocapnia and lowest at the highest CO₂ level in line with the finding that RR was largely reduced. In crabs acclimated to 4 °C, the RR was less affected by CO₂ such that activity caused lactate to increase with rising CO₂ levels. In line with the concept of oxygen and capacity limited thermal tolerance, all animals exposed to temperature extremes displayed a reduction in scope for performance, a trend exacerbated by increasing CO₂ levels. Additionally, the differences seen between cold- and warm-acclimated H. araneus after heat stress indicate that a small shift to higher acclimation temperatures also alleviates the response to temperature extremes, indicating a shift in the thermal tolerance window which reduces susceptibility to additional CO₂ exposure.