Mathematical lipid correction of δ13C and effect of lipid extraction on δ15N of European eel (Anguilla anguilla) muscle: Lipid correction of δ13C in European eels.

RATIONALE: Carbon (δ13C) and nitrogen (δ15N) stable isotope analysis is a powerful tool to investigate diverse questions in fish ecology, such as their trophic position or migration strategies. These questions appear particularly important to protect endangered European eel. However, elevated lipid content in eel muscle can bias δ13C values, as lipids are 13C-depleted compared to proteins and carbohydrates. METHODS: We measured δ13C and δ15N values of bulk and lipid-free samples of eel muscle. Lipid-free samples were obtained after the extraction of lipids with cyclohexane. Lipid-corrected δ13C values, using five different mathematical equations based on bulk δ13C values, were compared to lipid-free δ13C values. We also evaluated the effect of lipid extraction on δ15N values. The analyses were based on linear regression performed on 333 individuals captured in nine lagoons and four rivers. RESULTS: Independently to the capture site or habitat (river or lagoon), the predicted lipid-corrected δ13C values were highly consistent with the measured lipid-free δ13C values (R2 > 0.90). The application of specific equations for each habitat or capture site only slightly increases these R2 (1.5% or less). The lipid extraction treatment significantly decreased by 0.2‰ the δ15N values compared to bulk samples. CONCLUSIONS: Given the excellent prediction of mathematical equations and the small decrease of δ15N values after lipids extraction, we propose to use mathematical correction to estimate δ13C values of eel muscle. As the habitats or sites did not strongly influence the results, the coefficients from our study can be applied to other studies on European eel.

When more is more: taking advantage of species diversity to move towards sustainable aquaculture.

Human population growth has increased demand for food products, which is expected to double in coming decades. Until recently, this demand has been met by expanding agricultural area and intensifying agrochemical-based monoculture of a few species. However, this development pathway has been criticised due to its negative impacts on the environment and other human activities. Therefore, new production practices are needed to meet human food requirements sustainably in the future. Herein, we assert that polyculture practices can ensure the transition of aquaculture towards sustainable development. We review traditional and recent polyculture practices (ponds, recirculated aquaculture systems, integrated multi-trophic aquaculture, aquaponics, integrated agriculture-aquaculture) to highlight how they improve aquaculture through the coexistence and interactions of species. This overview highlights the importance of species compatibility (i.e. species that can live in the same farming environment without detrimental interactions) and complementarity (i.e. complementary use of available resources and/or commensalism/mutualism) to achieve efficient and ethical aquaculture. Overall, polyculture combines aspects of productivity, environmental protection, resource sharing, and animal welfare. However, several challenges must be addressed to facilitate polyculture development across the world. We developed a four-step conceptual framework for designing innovative polyculture systems. This framework highlights the importance of (i) using prospective approaches to consider which species to combine, (ii) performing integrated assessment of rearing environments to determine in which farming system a particular combination of species is the most relevant, (iii) developing new tools and strategies to facilitate polyculture system management, and (iv) implementing polyculture innovation for relevant stakeholders involved in aquaculture transitions.

TOFF, a database of traits of fish to promote advances in fish aquaculture.

Functional traits can be valuable pieces of information for aquaculture research and management. Although fish traits have been the focus of an abundant research, trait datasets for these organisms are difficult to access and often unpractical to achieve meta-analyses without a time-consuming extensive review. Already available large-scale compilations include trait information for many fish species but not as detailed as required for aquaculture purpose. Here, we introduce the TOFF (i.e. Traits OF Fish), a database focusing on fish functional traits that aims at bringing together behavioral, morphological, phenological, and physiological traits always coupled to environmental measurement context into a single open-source access repository. TOFF hosts data from published field and experimental studies. Here, we release data for 228 traits for 174 species extracted from 165 publications and present a collaborative platform. We ultimately aim at providing an inclusive and accessible data resource to facilitate advances in aquaculture development.

Effects of euthanasia methods on stable carbon (δ13 C value) and nitrogen (δ15 N value) isotopic compositions of fry and juvenile rainbow trout Oncorhynchus mykiss.

RATIONALE: Carbon and nitrogen stable isotope analyses of fish tissues are now commonly used in ecological studies but mostly require the sacrifice of the animal. Ethical considerations recommend the use of anesthetics for tissue sampling. This study examines how anesthetics affect stable isotope ratios of fish compared with other euthanasia methods. METHODS: Rainbow trout fry and juveniles were sacrificed using ice-freezing (as this common method used to kill fish does not affect natural isotopic ratios), electronarcosis or an overdose of chemical anesthetics (2-phenoxyethanol, benzocaine and clove oil). For fry, we sampled the whole animal whereas, for juveniles, white dorsal muscle, liver, red blood cells, plasma, external tegument and pectoral fin were sampled. Isotopic ratios and the elemental compositions of carbon and nitrogen were then measured. RESULTS: The δ15 N values, and the C and N contents of all considered tissues as well as δ13 C values of muscle, liver, red blood cells and plasma, were not affected by the use of chemical anesthetics. Clove oil and to a lesser extent 2-phenoxyethanol and benzocaine decreased δ13 C values of whole fry and juvenile external tegument and pectoral fin. The use of electronarcosis drastically affects the δ13 C and δ15 N values of all fish tissues. CONCLUSIONS: Anesthetics should be avoided for δ13 C analysis when tissues are in contact with the water containing the anesthetic. Ice-immersion has to be preferred when approved by guidelines. If not, benzocaine and 2-phenoxyethanol should be preferred over clove oil. Electronarcosis should not be used to kill fish until further investigations are performed.

Stable isotopes reveal spatial variability in the trophic structure of a macro-benthic invertebrate community in a tropical coral reef.

RATIONALE: Studies of organic matter fluxes in coral reefs are historically based on physical and biogeochemical approaches. It is important to link these approaches to community analysis as the abundance and behaviour of species, populations or trophic groups can have a profound effect on nutrient budgets. METHODS: We determined the carbon and nitrogen isotopic compositions of coral reef organic matter sources and macro-benthic invertebrate communities using a Europa Geo 20/20 isotope ratio mass spectrometer interfaced to an ANCA-SL elemental analyzer in continuous flow mode. Isotopic ecology metrics and a mixing model were used to analyze and interpret the data. RESULTS: The coral reef macro-invertebrate community principally relies on detrital or recycled food sources. An increased reliance on reef nitrogen-derived sources was observed in the cold-dry season. The community food-web lengths differ noticeably across the coral reef and reflect the characteristics and origin of organic matter reservoirs. CONCLUSIONS: Anthropogenic and terrestrial inputs lead to a loss of biological diversity. Exclusive dominance of suspension-feeding species is observed in areas receiving direct surface riverine particulate organic matter. The accumulation of sediment organic matter in eutrophic areas leads to dominance of deposit-feeding species. Copyright © 2016 John Wiley & Sons, Ltd.

Spatial and temporal variations in stable carbon (δ(13)C) and nitrogen (δ(15)N) isotopic composition of symbiotic scleractinian corals.

Tropical scleractinian corals are considered autotrophic as they rely mainly on photosynthesis-derived nutrients transferred from their photosymbionts. Corals are also able to capture and ingest suspended particulate organic matter, so heterotrophy can be an important supplementary trophic pathway to optimize coral fitness. The aim of this in situ study was to elucidate the trophic status of 10 coral species under contrasted environmental conditions in a French Polynesian lagoon. Carbon (δ(13)C) and nitrogen (δ(15)N) isotopic compositions of coral host tissues and photosymbionts were determined at 3 different fringing reefs during wet and dry seasons. Our results highlighted spatial variability in stable isotopic compositions of both coral host tissues and photosymbionts. Samples from the site with higher level of suspended particulate matter were (13)C-depleted and (15)N-enriched relative to corals and photosymbionts from less turbid sites. However, differences in both δ(13)C and δ(15)N between coral host tissues and their photosymbionts (Δ(host-photosymbionts 13)C and Δ(host-photosymbionts 15)N) were small (0.27 ± 0.76‰ and 1.40 ± 0.90‰, respectively) and similar at all sites, thus indicating no general increases in the heterotrophic pathway. Depleted δ(13)C and enriched δ(15)N values of coral host tissues measured at the most turbid site were explained by changes in isotopic composition of the inorganic nutrients taken up by photosymbionts and also by changes in rate of isotopic fractionation with environmental conditions. Our results also highlighted a lack of significant temporal variations in δ(13)C and δ(15)N values of coral host and photosymbiont tissues and in Δ(host-photosymbionts 13)C and Δ(host-photosymbionts 15)N values. This temporal stability indicated that corals remained principally autotrophic even during the wet season when photosymbiont densities were lower and the concentrations of phytoplankton were higher. Increased coral heterotrophy with higher food availability thus appears to be species-specific.

Can UV radiation affect benthic deposit-feeders through biochemical alteration of food resources? An experimental study with juveniles of the benthic polychaete Eupolymnia nebulosa.

The growth, tentacle development and feeding activity of the benthic polychaete Eupolymnia nebulosa were examined to determine whether UV might affect marine deposit-feeders indirectly through the modification of the nutritional quality of their resources. Since marine invertebrates have higher nutritional requirements during the period following settlement, we tested the effect of UV-altered phytodetritus on freshly settled juveniles of E. nebulosa. Phytodetritus was prepared from cultures of the diatom Skeletonema costatum either grown under or sheltered from UVB radiation. Sterol content of phytodetritus was unmodified by UV radiation. Conversely, phytodetritus was noticeably depleted in polyunsaturated fatty acids. Growth and tentacle development of juveniles fed on altered phytodetritus were reduced by 35% and 15% respectively, suggesting potential deficiencies in essential nutrients. In response to the lower quality of the phytodetritus, juveniles explored a wider area as they search for food, a strategy that could compensate for low food quality.

Sensitivity to UV radiation in early life stages of the Mediterranean sea urchin Sphaerechinus granularis (Lamarck).

The sea urchin Sphaerechinus granularis was used to investigate the impact of relevant levels of UV-B radiation on the early life stages of a common Mediterranean free spawning benthic species. Sperm, eggs and embryos were exposed to a range of UV radiation doses. The resulting endpoints were evaluated in terms of fertilisation success, development and survival rates. Above a weighted UV radiation dose of 0.0029 kJ m(-2), fertilisation capability of irradiated sperm decreased rapidly. The exposure of the eggs to 0.0175 kJ m(-2) and more led to delayed and inhibited development with ensuing embryonic morphological abnormalities. One-day old larvae remained strongly sensitive to UV radiation as shown by the 50% decrease of the larval survival rate for a dose of 0.025 kJ m(-2) UVR. The elevated sensitivity of embryos to experimental UVR went along with a lack of significant amount of sunscreen compounds (e.g., mycosporine-like amino acids) in the eggs. The present results demonstrated that gamete viability and embryonic development may be significantly impaired by solar UV radiation in S. granularis, compromising in this way the reproduction of the species. Unless adaptive behavioural reproductive strategies exist, the influence of ambient UV radiation appears as a selective force for population dynamics of broadcast spawners in the shallow benthic Mediterranean environment.

Ultraviolet radiation induces structural and chromatin damage in Mediterranean sea-urchin spermatozoa.

There is growing concern about the effects of enhanced levels of solar ultraviolet radiation on the living components of the biosphere (i.e. cancer, loss of biodiversity and productivity, etc.). In shallow coastal environments, many benthic species release their gametes directly in the water column where fertilisation occurs and the planktonic larvae remain for several weeks. Any effects on these early life stages could significantly impair reproductive input or alter the fitness of the community. The purpose of this paper is to provide new insights into the mechanisms of UV toxicity on sea-urchin spermatozoa in a cytological context, and to address the question of the potential ecological consequences of the damage. The Mediterranean sea-urchin Sphaerechinus granularis (Lamarck) was chosen as a model to study the effects of ecologically relevant doses of UV-R on the spermatozoa of marine invertebrates. Structural damage was visualised by use of transmission electron microscopy and the single-cell gel electrophoresis (Comet) assay was used to assess chromatin integrity in spermatozoa. The present results provide experimental evidence that irradiation with UV induces structural and chromatin damage in sea-urchin sperm. Almost 90% of spermatozoa exhibited morphological alterations and DNA strand breakage increased 2-fold. The observed alterations of the acrosome, plasma membrane and mitochondria can explain the concomitant impairment of fertilisation (23% decrease of fertilisation rate), which in turn may affect reproductive success. On the other hand, how DNA damage and fertilisation rate correlate remains unclear; however, when not repaired genetic lesions can lead to abnormal development and/or the transmission of heritable damage. The 3-fold decrease of the frequency of 2-celled embryos indicates a delay or inhibition of the first cell division, which may be ascribed to impairment of nuclear chromatin and/or other cellular targets.

Improved Comet assay for the assessment of UV genotoxicity in Mediterranean sea urchin eggs.

Gametes and embryos of broadcast spawners are exposed to a wide range of chemical and physical stressors which may alone, or in conjunction, have serious consequences on reproductive outcomes. In this study, two Mediterranean echinoid species, Paracentrotus lividus and Sphaerechinus granularis, were chosen as models to study the genotoxicity of UV radiation (UVR) on the eggs of broadcast-spawning marine invertebrates. The single cell gel electrophoresis, or Comet assay, was successfully adapted to assess DNA strand breakage in sea urchin eggs. The results demonstrated that the genetic material of sea urchin eggs is susceptible to environmentally realistic UV exposure. The induction of DNA damage in the irradiated unfertilized eggs suggests that the previously described defense mechanisms in sea urchin eggs do not completely protect the egg's DNA against UV toxicity. Taken together, our results suggest that UV-impairment of the genetic integrity of the eggs might have a role in postfertilization failures and abnormal embryonic development. Although both species were vulnerable to UVR, embryonic development was less dramatically impaired in P.Lividus. This observation supports the postulation that species inhabiting shallower environments possess more efficient mechanisms to overcome UV-induced DNA alterations. The present demonstration of the utility and sensitivity of the Comet assay to evaluate DNA integrity in eggs from marine invertebrates opens new perspectives for monitoring the long-term effects of environmental exposure on populations and for the routine screening of substances for genotoxicity in marine systems.