Use of stable isotopes to distinguish farmed from wild Atlantic salmon, Salmo salar

Abstract –  Stable isotopes of carbon and nitrogen ( δ ¹³C and δ ¹⁵N) were examined in wild and aquaculture origin Atlantic salmon, Salmo salar , to evaluate their utility to identify escaped farmed fish. Samples of muscle tissue obtained from wild Conne River, Newfoundland, salmon were significantly more enriched in nitrogen ( δ ¹⁵N: mean = 12.75; SD ± 0.38‰) but depleted in lipid corrected carbon ( δ ¹³C': mean = −20.51; SD ± 0.23‰) by comparison with aquaculture specimens obtained from Bay d'Espoir, Newfoundland ( δ ¹⁵N = 10.96 ± 0.19‰;  δ ¹³C' = −19.25 ± 0.17‰) resulting in a complete separation of the two groups. Aquaculture specimens differed in δ ¹³C' from analyses of commercial salmon diet by 0.24‰, within the enrichment range associated with trophic transfers, while the δ ¹⁵N values in salmon muscle were enriched by 5.01‰. Although differences occurred in direct comparisons of white muscle and adipose tissue ( N  = 49), the average δ ¹³C' and δ ¹⁵N signatures varied in absolute amounts by only 0.5‰, supporting the use of adipose tissue as a nonlethal means to determine isotopic signatures of Atlantic salmon.     

Basin geochemistry and isotopic ratios of fishes and basal production sources in four neotropical rivers

Abstract –  We analysed stable carbon and nitrogen isotopic ratios of dissolved inorganic carbon (DIC), plants, detritus and fishes to estimate the relative importance of dominant production sources supporting food webs of four Venezuelan rivers with divergent geochemical and watershed characteristics. Based on samples taken during the dry season at each site, fishes from two nutrient-poor, blackwater rivers had significantly lower δ ¹³C values (mean = −31.4‰ and −32.9‰) than fishes from more productive clearwater and whitewater rivers (mean = −25.2‰ and −25.6‰ respectively). Low carbon isotopic ratios of fishes from blackwaters were likely influenced by low δ ¹³C of DIC assimilated by aquatic primary producers. Although floodplains of three savanna rivers supported high biomass of C₄ grasses, relatively little carbon from this source appeared to be assimilated by fishes. Most fishes in each system assimilated carbon derived mostly from a combination of microalgae and C₃ macrophytes, two sources with broadly overlapping carbon isotopic signatures. Even with this broad overlap, several benthivorous grazers from blackwater and whitewater rivers had isotopic values that aligned more closely with algae. We conclude that comparative stable isotopic studies of river biota need to account for watershed geochemistry that influences the isotopic composition of basal production sources. Moreover, isotopic differences between river basins can provide a basis for discriminating spatial and temporal variation in the trophic ecology of fishes that migrate between watersheds having distinct geochemical characteristics.     

Natural abundance of 15N and 13C in fish tissues and the use of stable isotopes as dietary protein tracers in rainbow trout and gilthead sea bream

For developing efficient diets, two sets of experiments examined whether the use and allocation of dietary protein can be traced by labelling with stable isotopes (¹⁵N and ¹³C) in two culture fish ( Oncorhynchus mykiss and Sparus aurata) . In the first experiment, natural abundance and tissue distribution of these isotopes were determined, by measuring the δ¹³C and δ¹⁵N values by isotopic ratio mass spectrometry, in fingerlings (14–17 g) adapted to diets differing in the percentage of fish meal replacement by plant protein sources. For both species, δ¹⁵N and δ¹³C were greater in tissues with higher protein and lower lipid content. Delta ¹⁵N of diets and tissues decreased as replacement increased, suggesting δ¹⁵N can be used as a marker for dietary protein origin. The ¹⁵N fractionation (δ¹⁵N fish − δ¹⁵N diet) differed between groups, and could thus be used to indicate protein catabolism. In the second experiment, fish (75–90 g) of each species ingested a diet enriched with ¹⁵N-protein (10 g kg⁻¹ diet) and ¹³C-protein (30 g kg⁻¹ diet). These proportions were suitable for determining that the delta values of tissue components were high enough above natural levels to allow protein allocation to be traced at 11 and 24 h after feeding, and revealed clear metabolic differences between species.     

Evaluation of shrimp polyculture system in Thailand based on stable carbon and nitrogen isotope ratios

ABSTRACT: To quantify the contribution by cocultured animals to waste assimilation in an intensive shrimp farm in Thailand, the food web structures of the macrobenthos in a reservoir pond, a shrimp culture pond and water treatment ponds were examined using the stable C and N isotope ratio technique. Seawater for aquaculture was drawn from a creek, and stored in a reservoir pond, used for farming the banana prawn Fenneropenaeus merguiensis in culture ponds, and then recycled through treatment ponds where the green mussel Perna viridis was cultured to remove organic wastes discharged from the farming. The clam worm Nereididae sp. and the mud creeper Cerithideopsilla cingulata in the culture pond had δ ¹³C values of −21.0‰ and −18.4‰, respectively, suggesting that shrimp feed (mean δ ¹³C = −20.7‰) was the main food source for these species. The δ ¹³C analysis also suggested that sediments (−23.7‰) in the reservoir pond and particulate organic matter (POM) (−24.0‰) and/or sediments (−25.0‰) in the treatment pond supplied carbon for most macrobenthic animals. However, green mussels in the treatment pond had a mean δ ¹³C value of −20.5‰, suggesting that shrimp feed was the main food source for this species.     

Effects of lipid extraction on stable carbon and nitrogen isotope analyses of fish tissues: potential consequences for food web studies

Abstract –  We examined whether solvent-based lipid extractions, commonly used for stable isotope analysis (SIA) of biota, alters δ ¹⁵N or δ ¹³C values of fish muscle tissue or whole juvenile fish. Lipid extraction from muscle tissue led to only small (<1‰) isotope shifts in δ ¹³C and δ ¹⁵N values. By contrast, ecologically significant shifts (+3.4‰ for δ ¹³C and +2.8‰ for δ ¹⁵N) were observed for whole juvenile fish. Sample variance was not affected by lipid extraction. For tissue-specific SIA, two sample aliquots may be required: a lipid-extracted aliquot for stable carbon isotope analysis when differing lipid content among tissues is a concern, and a nonextracted aliquot for δ ¹⁵N determination. Whole organism SIA is not recommended because of the mix of tissues having different turnover times; for very small fish, we recommend that fish be eviscerated, decapitated, and skinned to minimise differences with samples of muscle tissue.     

Community and trophic structures of abalone <Emphasis Type="Italic">Haliotis diversicolor</Emphasis> habitat in Sagami Bay,Japan

Abstract:   The community and trophic structures of the subtidal habitat of the abalone Haliotis diversicolor were examined in Nagai, Sagami Bay, Japan. Conventional community indices showed no significant differences among three sampling stations. The overall continuum of stable isotope ratios was structured into three different trophic linkages: (i) brown algae-dependent benthic food chain; (ii) red algae-dependent benthic food chain; and (iii) planktonic food chain. Brown algae and red algae likely play different roles with respect to carbon sources in the habitat. Conventional fractionation values indicated that the abalone H. diversicolor (δ¹³C = −12.4 ± 1.0‰, δ¹⁵N = 9.3 ± 0.5‰) feeds on the lamina of Undaria pinnatifida during juvenile and adult stages (8.0–65.6 mm shell length). Stable isotope signatures suggested that the juveniles of other abalone species as well as some amphipods and a sea cucumber Holothuria decorata are competitors of H. diversicolor , whereas some Muricidae gastropods such as Thais bronni and Ergalatax contractus are predators. The isotopic differences among macroalgal species and the subsequent transfer to consumers indicate that stable isotopic analysis is an effective means of studying food webs in an open rocky shore community with little influence from external primary production such as terrestrial vegetation.     

Utilization of terrestrial organic matter by the bivalve <Emphasis Type="Italic">Corbicula japonica</Emphasis> estimated from stable isotope analysis

ABSTRACT:   Carbon and nitrogen isotopic ratios in tissue of the bivalve corbicula ( Corbicula japonica ) and particulate organic matter (POM) were measured along a salinity gradient in the Kushida Estuary, Japan. The bivalve exhibited a gradual isotopic enrichment from the uppermost estuarine site (δ¹³C = −24.8‰ and δ¹⁵N = 8.6‰) to the marine site (δ¹³C = −16.1‰ and δ¹⁵N = 11.8‰). Using the concentration-weighted mixing model, the bivalves' food source is estimated from the isotope values for the bivalves and POM from terrestrial plants, marine phytoplankton and benthic microalgae. The results indicated that the contributions of benthic micro algae and phytoplankton were small, while terrestrial particulate matter is significantly important for the corbicula diet, although the contribution varies among sampling sites.     

Ecological and genetic differentiation among the Arctic charr of Lake Aigneau, Northern Québec

Abstract –  Two modal size groups of sexually mature Arctic charr ( Salvelinus alpinus ) differing in shape and found at different depths in Lake Aigneau in the Canadian sub-Arctic are described and tested for genetic and ecological differentiation. Forms consisted of a small littoral resident, mean size 21.7 cm, and a large profundal resident, mean size 53.9 cm. Mitochondrial DNA analysis indicated that seven of eight haplotypes were diagnostic for either the littoral or profundal fish, with 66.6% of the variation being found within form groupings. Pairwise tests of microsatellite data indicated significant differences in nine of 12 loci and a significant difference between the forms across all tested loci. Molecular variation was partitioned to 84.1% within and 15.9% between forms and suggestive of either restricted interbreeding over time or different allopatric origins. Stable isotope signatures were also significantly different, with the profundal fish having higher δ¹³C and δ¹⁵N values than the littoral fish. Overlap and separation, respectively, in the range of form δ¹³C and δ¹⁵N signatures indicated that carbon was obtained from similar sources, but that forms fed at different trophic levels. Littoral fish relied on aquatic insects, predominantly chironomids. Profundal fish were largely piscivorous, including cannibalism. Predominantly empty stomachs and low per cent nitrogen muscle-tissue composition among profundal fish further indicated that the feeding activity was limited to the winter when ice-cover increases the density of available prey at depth. Results provide evidence of significant differences between the modal groups, with origins in both genetics and ecology.     

Patterns of stable carbon and nitrogen isotopes in the baleen of common minke whale Balaenoptera acutorostrata from the western North Pacific

ABSTRACT:   Stable carbon and nitrogen isotope ratios were determined in the baleen plates of 17 common minke whales Balaenoptera acutorostrata from the north-western Pacific Ocean off Japan, as well as prey species (krill Euphausia pacifica , Japanese anchovy Engraulis japonicus and Pacific saury Cololabis saira ) collected in the stomach contents, to investigate the trophic relationship between the minke whales and their prey. A few δ¹⁵N-depleted peaks occurred along the length of baleen plates for 10 males irrespective of stomach content (anchovies and sauries). Similar δ¹⁵N-depleted peaks were also found for one female and two immature individuals. It was likely that these δ¹⁵N-depleted peaks formed in early summer. The stable nitrogen isotope ratio (δ¹⁵N) values in Pacific saury (9.3 ± 1.4‰) did not differ significantly from that in Japanese anchovy (8.8 ± 0.9‰). In contrast, δ¹⁵N in krill (7.2 ± 0.5‰ in July and 8.0 ± 0.2‰ in September) were significantly lower than in the Pacific saury. Thus, these peaks may reflect the dietary change from krill to fishes in the feeding migration of the whales. Growth rate of the baleen plate was estimated to be 129 mm/y, and it appeared that a dietary record of about 1.4 years remained in the baleen plate. For two immature whales, the maximum value of δ¹⁵N occurred at the tip of baleen. This δ¹⁵N enrichment may possibly be useful for discriminating weanlings and older whales.     

Comparing trophic position of stream fishes using stable isotope and gut contents analyses

Abstract –  Stable isotope analysis (SIA) and gut contents analysis (GCA) are commonly used in food web studies, but few studies analyse these data in concert. We used SIA (δ¹⁵N) and GCA (% composition) to identify diets and trophic position (TP) of six stream fishes and to compare TP estimates between methods. Ordination analysis of gut contents identified two primary trophic groups, omnivores and predators. Significant differences in TP_GCA and TP_SIA were similar in direction among-species and among-trophic groups; neither method detected seasonal changes in omnivore diets. Within-species TP_GCA and TP_SIA were similar except for one omnivore. TP_GCA was less variable than TP_SIA for predators, but variation between methods was similar for omnivores. While both methods were equally robust at discriminating trophic groups of fishes, TP_SIA is less laborious to estimate and may facilitate cross-stream comparisons of food web structure and energy flow.     

Stable isotope analysis of trophic position and terrestrial vs. marine carbon sources for juvenile Pacific salmonids in nearshore marine habitats

Abstract  Stable isotope analysis was used to determine trophic position and the relative contributions of terrestrial-derived carbon (TDC) and marine-derived carbon for Chinook, Oncorhynchus tshawytscha (Walbaum), pink, Oncorhynchus gorbuscha (Walbaum), and chum, Oncorhynchus keta (Walbaum) salmon fry in near-shore marine habitats. Chum fry were enriched in δ ¹³C relative to pink fry, and enriched in δ ¹⁵N relative to both Chinook and pink fry. Between 5.5 and 39.7% of the carbon in the three species was TDC. The TDC was higher in chum fry (28.7 ± 4% SD) than in pink fry (24.9 ± 4.4% SD), but TDC did not differ between Chinook fry (27.8 ± 9.5% SD) and either chum or pink fry. The fry of these three species of Pacific salmon may form a trophic hierarchy with chum fry occupying the highest trophic position and the three species may also partition resources according to carbon source.     

Carbon sources of Amazonian fisheries

Variation in the seasonal and spatial isotopic composition of plant C₄ (aquatic macrophytes) and C₃ (forest, C₃ aquatic macrophytes and algae), and that of fish [ Prochilodus nigricans Agassiz, Mylossoma duriventre (Cuvier), Colossoma macropomum (Cuvier), Semaprochilodus insignis (Schomburgk) and S. taeniurus Steindachner in the Amazon floodplain were analysed to test whether the fisheries deliver plant carbon to the population of Manaus in the same proportion as it is available in the floodplain. The contribution of C₄ plants was significantly lower in ¹³C during the season of high water levels and increased toward the west of the basin. Mylossoma duriventre and C. macropomum changed δ¹³C levels, while the δ¹³C of P. nigricans and C. macropomum shifted spatially. The contribution of C₄ to the fisheries yield was small. C³ plants (excluding phytoplankton) also contributed less than expected. This was explained by the importance of detritivores to the yield of the fisheries and the dependence of these species on algal carbon.     

Stable carbon and nitrogen isotopic evidence for ecotonal coupling between boreal forests and fishes

Abstract— As a result of water turbulence effects on boundary layer diffusion resistance and carbon isotopic discrimination, the δ¹³C values (ratios of ¹³C: ¹²C) of attached algae may often overlap those of terrestrial plants, thereby making it impossible to distinguish between the relative importance of these two potential food sources for aquatic animals. The present study used a dual isotope approach (δ¹³C and δ¹⁵N) to refine measurements of the incorporation of allochthonous organic matter into freshwater fishes. The dependence of five species of littoral fishes on terrestrial detritus for part of their energy sustenance was demonstrated. The littoral zones of boreal Canadian Shield lakes are, therefore, not isolated from their surrounding riparian forests in terms of carbon flow as present day timber management guidelines erroneously assume, but instead exhibit a measurable degree of ecotonal land-water coupling. As a result, clearcut logging of riparian forests to the lakeshore edge, permissible by law in most Canadian provinces containing boreal forests, may have to be reassessed as a forest harvesting strategy.     

Stable Isotopic Evaluation of the Relative Assimilation of Natural and Artificial Foods by Golden Shiners Notemigonus crysoleucas in Ponds

Stable carbon isotope ratios (δ¹³C) were used to estimate the relative contributions of natural and prepared feeds to the nutrition of golden shiners Notemigonus crysoleucas in ponds. Concurrent feeding trials were conducted in tanks (110 L) and ponds (0.04 ha) to determine the response of golden shiners to three isotopically distinct prepared diets both in the presence (pond) and absence (tank) of natural food. Seston was the primary indicator of the δ¹³C of natural foods available in the ponds, but δ¹³C's of free-swimming, unfed golden shiners provided an additional index. Fish receiving experimental diets were maintained in floating nets within the same ponds. After 6 wk, fish in tanks fed all three diets had acquired the diet label to some degree, while fish in ponds also tracked their respective feeds except for those fed diet 3. This diet contained a large amount of corn meal, which appeared to be poorly assimilated by golden shiners. The use of natural foods by golden shiners fed all three prepared feeds ranged from 40–83% and was inversely correlated with the relative assimilation of the prepared feeds.     

Invasion of stream fishes into low trophic positions

Abstract –  In the absence of other life-history constraints, fishes that can feed at low trophic levels (i.e., omnivores/detritivores) are predicted to be successful invaders because their food resources during the colonization and integration phases of the invasion are rarely limiting. Accordingly, we hypothesized that trophic position of non-native fishes in five mid-western and south-western U.S. river systems would be lower than native fishes. By standardizing δ ¹⁵N values with a common invertebrate (chironomids) across sites, we were able to evaluate differences in trophic position between natives and non-natives across sample locations and years. Our data tend to support this hypothesis, but there was notable spatial and temporal variation in this pattern. Moreover, three of four species generally fed at lower trophic positions in their introduced ranges than their native ranges. Although many factors influence the success of introduced species, our data suggest that the ability to forage on low-quality resources is a favourable trait for invasive fishes in lotic systems. Because these fishes feed at low trophic positions, it is important to consider how they will influence invaded systems, in contrast to invaders that feed at higher trophic positions. Future studies that evaluate the resistance of communities to invaders from different trophic positions would help understand the mechanisms that control the establishment and spread of species with different life-history traits.     

Linking migratory patterns and diet to reproductive traits in female brown trout (Salmo trutta L.) by means of stable isotope analysis on ova

Abstract –  Many populations of vertebrates display alternative reproductive strategies involving delays in maturation processes and migrations to more productive habitats. Notably for females, migratory patterns and diet could influence reproductive traits such as ova size and number. To test this hypothesis, we targeted anadromous and freshwater-resident female brown trout of various sizes during their return to spawn in tributaries, and carried out stable isotope analyses (δ¹⁵N and δ¹³C) on ova to identify their feeding areas and diet. The study was performed in the Oir River (France), a small coastal system where trout migrations to sea and within the river basin have been monitored for the last 20 years. General trends were observed in reproductive traits, such as the positive correlation between female fork length, fecundity and ova size. Fecundity was size-dependent whatever the migratory origin of females, but major divergences were seen for ova size and isotopic ratios. Freshwater residents displayed a constant increase in ova size and a ¹⁵N-enrichment in ova, suggesting an investment in larger ova and a shift to piscivory as the fish become larger and older. Conversely, anadromous females had smaller ova compared with freshwater-resident females of similar body size and achieved higher fecundity as they grew bigger. Our results suggest the existence of a continuum of reproductive traits for freshwater-resident females whereas anadromous females clearly show a break with this continuum. This major dissimilarity could be explained by the difference in growing environments in terms of food availability and quality.     

Pharmacokinetics of Dietary 13C-Labeled Docosahexaenoic Acid and Docosapentaenoic Acid in Red Sea Bream Chrysophrys major

The objectives of this study were to investigate: 1) the pharmacokinetics of dietary docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA) using ¹³C-labeled fatty acids; 2) the interorgan transport of DHA in the red sea bream by monitoring the DHA level of several organs; and 3) the relationship between the plasma DHA level and optimum dietary DHA level in the plasma of the red sea bream Chrysophrys major . For this purpose, a mixture of 38.5% of [¹³C]DHA, 8.5% of [¹³C]DPA, and 4.2% of [¹³C]palmitic acid were given to the red sea bream at dose level of 8.0, 16.0, and 47.9 mg/kg by a single oral administration. For [¹³C]DHA, the maximum plasma concentration (t_max) occurred at 2.00–3.00 h after the oral administration. The peak plasma concentration (C_max) and the area under the plasma concentration-time curve to 24 h (AUC_0-24 for [¹³C]DHA level linearly increased with respect to dosage. [¹³C]DHA appeared in each organ (plasma, erythrocyte and the fat body of the orbit, liver, intestine, skin, brain, heart and muscle) at 0.5 h and was observed until 24 h. From the values determined for the pharmacokinetic parameters, the range of the effective plasma DHA level for normal growth of the red sea bream was suggested to be between 21.0 and 40.3 μg/mL. For [¹³C]DPA, the AUC_0-24 and C_max values also linearly increased with the dosage, but t_max did not depend on it.     

Lipid biosynthesis from [14C]-glucose in the coral Montipora digitata

ABSTRACT:     Glucose has been implicated in functioning as a form of carbon translocated from symbiont zooxanthellae to the host coral cell. The present paper describes the lipid biosynthesis from [¹⁴C]-glucose in the coral tissue. To study the incorporation of [¹⁴C]-glucose into lipids, the branch tips of the coral Montipora digitata were incubated with [¹⁴C]-glucose or another radiolabeled substrates. The lipid biosynthesis from [¹⁴C]-glucose was dependent on light, and was decreased by dark conditions or by photosystem II inhibitor, 3-(3,4-dichlorophenyl)-1,1-dimethylurea. Of the lipid classes, the light dependency was more pronounced with the biosynthesis of triacylglycerols (TG) and wax compared with phospholipids. Examination of [¹⁴C]-label distribution in the glycerolipids suggested that [¹⁴C]-glucose supplied mainly the fatty acid moiety of newly synthesized TG, while [¹⁴C]-glucose provided evenly the fatty acid moiety and the glycerol skeleton of phospholipids. The comparison of [¹⁴C]-labeling of lipid from host coral tissue and symbiont zooxanthellae suggested that [¹⁴C]-glucose entered the coral cell and was processed in parallel in the zooxanthellae and host cells. Furthermore, the coral cells used various [¹⁴C]-labeled sugars for lipid synthesis with similar lipid labeling profile as was the case for glucose. The current study thus supports the view that the low-molecular-weight compound, sugars and amino acids, once translocated from zooxanthellae to host cell were metabolized toward lipogenesis as well as glycerol.     

Linear models to predict the digestible lipid content of fish diets

Values for the digestible contents of nutrients in diets and feed ingredients are of utmost importance in nutritional strategies for fish. Prediction from dietary composition would eliminate lengthy, tedious and demanding digestibility experiments with fish. Apparent digestible lipid (DL) content [range 7.6–353.4 g kg⁻¹ dry matter (DM)] in compound diets can be predicted with high accuracy ( n  = 610; studies =127; fish species = 34; R ² = 0.9515; RMSE = 16.9504) from dietary crude lipid (CL) content (range 12.0–388.7 g kg⁻¹ DM) by the linear regression equation DL =−2.7303 + 0.9123 CL. Validation of this equation against 65 values from 15 independent studies presented R ² and mean prediction error (MPE) values of 0.9947 and 0.0671, respectively. The corresponding equation for 37 individual feed ingredients evaluated in 24 studies with 18 fish species ( n  = 180) was found to be DL = −1.5824 + 0.8654 CL ( R ² = 0.9717; RMSE = 8.3765). However, validation of the latter is currently hampered by a lack of independent values.     

Effects of temperature, salinity and irradiance on the growth of the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae) isolated from Hiroshima Bay, Japan

ABSTRACT: Temperature and salinity ranges in which Gymnodinium catenatum (Hiroshima Bay strain) showed specific growth rates higher than 0.2/day were approximately 20–30°C and 20–32. The specific growth rate (μ), expressed as a polynomial equation as functions of temperature ( T ; °C) and salinity ( S ) were μ = (−6.84 × 10⁻⁴ T ² + 0.0354 T – 0.213) × (−1.03 × 10⁻³ S ² + 0.0579 S – 0.548)/0.31; the maximum growth rate (0.31/day) was obtained at 25°C and 30. From a comparison with field data recording temperature, salinity and light intensity, this species may be expected to bloom from summer to autumn in Hiroshima Bay.