Assessing the effects of lipid extraction and lipid correction on stable isotope values (δ13 C and δ15 N) of blubber and skin from southern hemisphere humpback whales.

RATIONALE: The coupled analysis of δ13 C and δ15 N stable isotope values of blubber and skin biopsy samples is widely used to study the diet of free-ranging cetaceans. Differences in the lipid content of these tissues can affect isotopic variability because lipids are depleted in 13 C, reducing the bulk tissue 13 C/12 C. This variability in carbon isotope values can be accounted for either by chemically extracting lipids from the tissue or by using mathematical lipid normalisation models. METHODS: This study examines (a) the effects of chemical lipid extraction on δ13 C and δ15 N values in blubber and skin of southern hemisphere humpback whales, (b) whether chemical lipid extraction is more favourable than mathematical lipid correction and (c) which of the two tissues is more appropriate for dietary studies. Strategic comparisons were made between chemical lipid extraction and mathematical lipid correction and between blubber and skin tissue δ13 C and δ15 N values, as well as C:N ratios. Six existing mathematical normalisation models were tested for their efficacy in estimating lipid-free δ13 C for skin. RESULTS: Both δ13 C and δ15 N values of lipid-extracted skin (δ13 C: -25.57‰, δ15 N: 6.83‰) were significantly higher than those of bulk skin (δ13 C: -26.97‰, δ15 N: 6.15‰). Five of the six tested lipid normalisation models had small error terms for predicting lipid-free δ13 C values. The average C:N ratio of lipid-extracted skin was within the lipid-free range reported in other studies, whereas the average C:N ratio of blubber was higher than previously reported. CONCLUSIONS: These results highlight the need to account for lipids when analysing δ13 C and δ15 N values from the same sample. For optimised dietary assessments using parallel isotope analysis from a single sample, we recommend the use of unextracted skin tissue. δ15 N values should be obtained from unextracted skin, whereas δ13 C values may be adequately lipid corrected by a mathematical correction.

Identification of compounds from terrestrial dissolved organic matter toxic to cyanobacteria.

There is emerging evidence for the phytotoxicity of terrestrial dissolved organic matter (DOM), however its sources, transformations and ecological effects in aquatic ecosystems are poorly understood. DOM characterization by Nuclear Magnetic Resonance (NMR) spectroscopy has typically involved solid-state techniques, but poor resolution has often precluded identification of individual components. This study is the first to directly identify individual phytotoxic components using a novel combined approach of preparative HPLC fractionation of DOM (obtained from leaves of two common riparian trees, Casuarina cunninghamiana and Eucalyptus tereticornis). This was followed by chemical characterization of fractions, using one-dimensional (1D) and two-dimensional (2D) solution-state 1H NMR analyses. Additionally, the phytotoxic effect of the fractions was determined using cultures of the cyanobacteria Raphidiopsis (Cylindrospermopsis) raciborskii. The amino acid, proline, from Casuarina leachate was identified as phytotoxic, while for Eucalyptus leachate, it was gallic acid and polyphenols. These phytotoxicants remained in the leachates when they were incubated in sunlight or the dark conditions over 5 days. Our study identifies phytotoxic compounds with the potential to affect algal species composition, and potentially control nuisance R. raciborskii blooms.

Subtropical freshwater phytoplankton show a greater response to increased temperature than to increased pCO2.

Global increases in atmospheric CO2 and temperatures will impact aquatic systems, with freshwater habitats being affected. Some studies suggest that these conditions will promote cyanobacterial dominance. There is a need for a clearer picture of how algal species and strains within species will respond to higher temperatures and CO2, especially in combination. This study examined two chlorophytes (Monoraphidium and Staurastrum), and two strains of the cyanobacterium Raphidiopsis raciborskii (straight S07 and coiled C03), to determine how the combination of higher temperature and CO2 levels will affect their growth and maximum cell concentrations. Continuous cultures were used to compare the steady state cell concentrations at 28 °C and 30 °C, and CO2 partial pressures (pCO2), 400 and 750 ppm for all cultures, and in addition 1000 ppm at 28 °C for R. raciborskii strains. This study showed that, for all species, water temperature had a greater effect than higher pCO2 on cell concentrations. There were clear differences in response between the chlorophyte species, with Monoraphidium preferring 28 °C and Staurastrum preferring 30 °C. There were also differences in response of the R. raciborskii strains to increasing temperature and pCO2, with S07 having a greater increase in cell concentration. Genome analysis of R. raciborskii showed that the straight strain has five additional carbon acquisition genes (ß-CA, chpY, cmpB, cmpD and NdhD4), indicative of increased carbon metabolism. These differences in the strains' response to elevated pCO2 will lead to changes in the species population structure and distribution in the water column. This study shows that it is important to examine the effects of both pCO2 and temperature, and to consider strain variation, to understand how species composition of natural systems may change under future climate conditions.

Uptake and accumulation of cadmium, manganese and zinc by fisheries species: Trophic differences in sensitivity to environmental metal accumulation.

Fishery targeted species living in estuaries face multiple anthropogenic pressures including habitat contamination. However, trace metal concentrations in aquatic organisms can be highly variable, making it difficult to interpret accumulation responses. Understanding sources for metal accumulation in these organisms and their biokinetics is important for management of local fisheries and ensuring safety and quality of consumed seafood, particularly in urbanised areas. In this study, we exposed Australian sand clams, school prawns and sand whiting to a combination of cadmium (Cd), manganese (Mn) and zinc (Zn) radioisotopes 1) dissolved in seawater, 2) adsorbed to suspended sediment particles and 3) in radiolabelled food. Sand clams were sensitive to Cd, Mn and Zn uptake and accumulation from all sources because of their filter feeding physiology. Mean Cd and Zn assimilation efficiencies (AE) were higher in clams fed benthic diatoms (51, 43, 63% for Cd, Mn and Zn, respectively) than clams fed an algal flagellate species (22, 32, 33% for Cd, Mn and Zn, respectively). Metal uptake by prawns from seawater was low, whereas assimilation from diet was high (67, 59, 64% mean AEs from Cd, Mn and Zn, respectively). Sand whiting did not accumulate metals from seawater, even after concentrations were increased. Assimilation from diet (labelled prawns) was also low for sand whiting, particularly for Cd and Zn (11, 26, 14% mean AEs from Cd, Mn and Zn, respectively). These results may help explain the persistence of sand whiting in contaminated estuaries. Suspended sediment exposures showed that prawns and fish are less likely than clams to be negatively affected by disturbance events such as floods, which can bring metals into estuaries. The findings of this study have implications for fisheries management, both for protection and remediation of important habitats, and to ensure safe standards for seafood consumption by humans.