Paleosalinity in a brackish lake during the Holocene based on stable oxygen and carbon isotopes of shell carbonate in Nakaumi Lagoon, southwest Japan

Based on the relationship between salinity and δ¹⁸O and δ¹³C of modern shells in the Lake Nakaumi-Shinji lagoon system (southwestern Japan), where the salinity changes regularly from ca. 1 PSU to 34 PSU, a paleosalinity record for Nakaumi Lagoon during the Holocene has been derived from bulk mollusk shell δ¹⁸O and δ¹³C data. The robust relationships between the salinity and modern shell δ¹⁸O_ar and δ¹³C_ar (aragonite) were used to calibrate the paleosalinity reconstruction. The salinity relationships are expressed by the regressions:

Salinity (PSU)=3.86 δ¹⁸O_ar(‰ VPDB)+33.9 (n=18, r=0.978)     

Oxygen and carbon isotope compositions of middle Cretaceous vertebrates from North Africa and Brazil: Ecological and environmental significance

In order to investigate mid-Cretaceous terrestrial climates of low paleolatitudes, Moroccan, Tunisian and Brazilian vertebrate apatites have been analyzed for their oxygen and carbon isotope compositions of phosphates (δ¹⁸O_p) and carbonates (δ¹⁸O_c, δ¹³C_c). At each site, coexisting theropod dinosaurs, titanosaurid sauropods, pterosaurs, crocodilians, turtles and fish have distinct δ¹⁸O_p and δ¹³C_c values reflecting their ecologies, diets and foraging environments. Oxygen isotope compositions of surface waters (δ¹⁸O_w) estimated from turtle and crocodile δ¹⁸O_p values range from − 5.0 ± 1.0‰ to − 2.4 ± 1.0‰, which do not differ from mean annual rainwater values occurring today under inter-tropical sub-arid to arid climates. High water temperatures ranging from 21 ± 6 °C to 34 ± 2 °C deduced from fish δ¹⁸O_p values are in agreement with those published for mid-Cretaceous low latitudes. Temporary or seasonal droughts are inferred from high δ¹⁸O_p values of lungfish teeth, even though lower reptile δ¹⁸O_p values suggest the use of distinct and most likely larger or regularly renewed bodies of water. Environmental conditions of the studied low latitude regions during the Aptian-Cenomanian interval were somewhat similar to those experienced today under semi-arid to arid tropical or equatorial climates, but with higher mean surface temperatures than present-day ones.     

Effects of body size and environment on diet-tissue δC fractionation in fishes

δ¹³C data are often used in trophodynamic research where diet-tissue fractionation (Δδ¹³C) is assumed to be 0-1‰ per trophic level and unaffected by the size of animals or their environment. Variation in Δδ¹³C will influence conclusions about food sources, energy pathways and trophic level. To assess the effects of body size, age and environmental conditions on Δδ¹³C, European sea bass (Dicentrarchus labrax) were reared on constant diets of dab (Limanda limanda) or (Ammodytes marinus) for 2years under natural environmental regimes. Bass were sampled approximately monthly to determine Δδ¹³C for muscle, heart and liver tissue and were 1.66‰, − 0.18‰, − 1.77‰ (sandeel diet) and 1.34‰, − 1.18‰, − 1.75‰ (dab diet) respectively. Arithmetic lipid correction increased Δδ¹³C to > 2‰ for muscle and liver. Δδ¹³C was dependent on body mass and experimental duration (age) and generally declined with weight or time even after correction for lipid content. For liver, increasing temperature increased Δδ¹³C. The Δδ¹³C estimates from this study were compared with all available published Δδ¹³C estimates for fish. Bass muscle Δδ¹³C was similar to previous estimates for fish white muscle Δδ¹³C (1.56 ± 1.10‰) and whole body Δδ¹³C (1.52 ± 1.13‰). Fractionations derived in this study, combined with those from the literature, support the use of diet-tissue fractionation values of between 1‰-2‰ for δ¹³C, rather than the commonly used 0‰ − 1‰. For muscle Δδ¹³C, 1.5‰ is appropriate.     

Variation in bulk tissue, fatty acid and monosaccharide δ13C values between autotrophic and heterotrophic plant organs

The flowers of 23 species of grass and herb plants were collected from a mesotrophic grassland to assess natural variability in bulk, monosaccharide and fatty acid δ¹³C values from one plant community and were compared with previous analyses of leaves from the same species. The total mean bulk δ¹³C value of flower tissues was −28.1‰, and there was no significant difference between the mean δ¹³C_flower values for grass (−27.8‰) and herb (−28.2‰) species. On average bulk δ¹³C_flower values were 1.1‰ higher than bulk δ¹³C_leaf values, however, the δ¹³C_flower and δ¹³C_leaf values of grasses did not differ between organs suggesting that carbon isotope discrimination is different in grass and herb species. The abundance of different monosaccharides abundance varied between plant types, i.e. xylose concentrations in the grass flowers were as high as 40%, compared with up to 15% in the herb species, but the general relationship δ¹³C_arabinose > δ¹³C_xylose > δ¹³C_glucose > δ¹³C_galactose which had been observed in leaves was similar in flowers (total mean δ¹³C values = −25.9‰, −27.2‰, −28.8‰ and −28.1‰, respectively). However, the average 5.4‰ depletion in the δ¹³C values of the C_16:0, C_18:2 and C_18:3 fatty acids in flowers compared to bulk tissue was significantly greater than observed for leaves. The trend C_16:0 < C_18:2 < C_18:3 previously observed in leaves was also observed in grass flowers (δ¹³C_C16:0 = −33.8‰; δ¹³C_C18:2 = −33.1‰; δ¹³C_C18:3 = −34.2‰) but not herb flowers (δ¹³C_C16:0 = −34.1‰; δ¹³C_C18:2 = −32.4‰; δ¹³C_C18:3 = −34.5‰). We conclude: (i) that the biological processes influencing carbon isotope discrimination in grass flowers are different from herbs flowers; and, (ii) that a range of post-photosynthetic fractionation effects caused the observed differences between flower and leaf δ¹³C values, especially the significant ¹³C-depletion in flower fatty acid δ¹³C values.     

Seasonal variations in bulk tissue, fatty acid and monosaccharide δC values of leaves from mesotrophic grassland plant communities under different grazing managements

Leaves of 26 grass, herb, shrub and tree species were collected from mesotrophic grasslands to assess natural variability in bulk, fatty acid and monosaccharide δ¹³C values under different grazing management (cattle- or deer-grazed) on three sample dates (May, July and October) such that interspecific and spatiotemporal variations in whole leaf tissues and compound-specific δ¹³C values could be determined. The total mean leaf bulk δ¹³C value for plants was −28.9‰ with a range of values spanning 7.5‰. Significant interspecific variation between bulk leaf δ¹³C values was only determined in October (P = <0.001) when δ¹³C values of the leaf tissues from both sites was on average 1.5‰ depleted compared to during July and May. Samples from May were significantly different between fields (P = 0.03) indicating an effect from deer- or cattle-grazing in young leaves. The average individual monosaccharide δ¹³C value was 0.8‰ higher compared with whole leaf tissues. Monosaccharides were the most abundant components of leaf biomass, i.e. arabinose, xylose, mannose, galactose and glucose, and therefore, fluctuations in their individual δ¹³C values had a major influence on bulk δ¹³C values. An average depletion of ca. 1‰ in the bulk δ¹³C values of leaves from the deer-grazed field compared to the cattle-grazed field could be explained by a general depletion of 1.1‰ in glucose δ¹³C values, as glucose constituted >50% total leaf monosaccharides. In October, δ¹³C values of all monosaccharides varied between species, with significant variation in δ¹³C values of mannose and glucose in July, and mannose in May. This provided an explanation for the noted variability in the tissue bulk δ¹³C values observed in October 1999. The fatty acids C_16:0, C_18:2 and C_18:3 were highly abundant in all plant species. Fatty acid δ¹³C values were lower than those of bulk leaf tissues; average values of −37.4‰ (C_16:0), −37.0‰ (C_18:2) and −36.5‰ (C_18:3) were determined. There was significant interspecific variation in the δ¹³C values of all individual fatty acids during October and July, but only for C_18:2 in May (P = <0.05). This indicated that seasonal trends observed in the δ¹³C values of individual fatty acids were inherited from the isotopic composition of primary photosynthate. However, although wide diversity in δ¹³C values of grassland plants ascribed to grazing management, interspecific and spatiotemporal influences was revealed, significant trends (P = <0.0001) for fatty acid and monosaccharide δ¹³C values: δ¹³C_16:0 < δ¹³C_18:2 < δ¹³C_18:3 and δ¹³C_arabinose > δ¹³C_xylose > δ¹³C_glucose > δ¹³C_galactose, respectively, previously described, appear consistent across a wide range of species at different times of the year in fields under different grazing regimes.     

δN and δC analysis of a Posidonia sinuosa seagrass bed

Potential food sources and dominant invertebrates and fishes were collected for the examination of variability in ¹³C/¹²C and ¹⁵N/¹⁴N to determine the sources of carbon available to consumers within a Western Australian Posidonia sinuosa-dominated seagrass bed. Autotrophs showed a wide distribution of δ¹³C values, with P. sinuosa at −11.3 ± 0.8‰ and macroalgae ranging from −16.6 to −31.7‰. This variation allowed us to successfully identify macroalgae as the main contributor of carbon to the trophic structure, although no distinction could be made between epiphytic macroalgae on seagrass, or allochthonous macroalgal sources. The range in δ¹⁵N ratios among potential food items at the trophic base was too small to make it useful as tracer of nitrogen flow pathways, but it consistently increased from macrophytes and detritus (4.1–6.8‰), to invertebrates (5.7–7.4‰) located near the middle of the food web, to fishes (8.3–11.9‰), with piscivorous species such as Leviprora inops generally having a higher ¹⁵N. δ¹³C of seston (−12.8‰) and sedimentary organic matter (−8.7‰) indicate that seagrass material is the main contributor to these two carbon pools, and that very little of it contributes to animal biomass.     

Stable isotope fractionation in the digestive gland, muscle and gills tissues of the marine mussel Mytilus galloprovincialis

Mytilus galloprovincialis is a common species in the Mediterranean. It is a sedentary filter-feeding organism that assimilates carbon and nitrogen isotopic ratios in tissues from its food sources. The δ¹³C and δ¹⁵N values have been used to demonstrate differences in isotopic composition between digestive gland, muscle and gills of this mussel. For δ¹³C, mean values were - 21.99 ± 0.50‰, - 19.70 ± 0.44‰, and - 19.96 ± 0.44‰, respectively; and for δ¹⁵N, they were 5.16 ± 0.90‰, 7.67 ± 0.79‰ and 7.77 ± 0.85‰, respectively. The fractionation or enrichment factor for ¹³C values between digestive gland and muscle, between digestive gland and gills, and between muscle and gills were - 2.29 ± 0.16‰, - 2.04 ± 0.14‰ and 0.27 ± 0.07‰, respectively, within the expected range of ¹³C fractionation at filter feeders reported elsewhere. In contrast, low fractionation values were found for ¹⁵N with - 2.45 ± 0.24‰, - 2.51 ± 0.16‰ and - 0.11 ± 0.16‰, between digestive gland and muscle, between digestive gland and gills, and between muscle and gills, respectively. Through isotopic fractionation of M. galloprovincialis, the depleted values were found in the digestive gland, followed by gills and then by muscle tissue. Statistical analysis (PERMANOVA) was performed to check for significant differences in δ¹³C and δ¹⁵N isotopic signatures between tissues and localities. The current study demonstrates significant differences in the δ¹³C and δ¹⁵N isotopic composition between digestive gland, muscle and gills tissues in M. galloprovincialis living in the oligotrophic environment of the Balearic Islands.     

Hominid environments at Hadar from paleosol studies in a framework of Ethiopian climate change

The amount and seasonal distribution of paleo-rainfall is a major concern of paleoanthropology because they determine the nature of the vegetation and the structure of the ecosystem, particularly in eastern Africa. The δ¹⁸O and δ¹³C of paleosol carbonates are quantitative proxies of these critical features of the paleoenvironment. The Afar region of Ethiopia lies between the African and Indian summer monsoons, and is prone to profound climate change. In the western Afar, the dominant paleoenvironment of the Hadar Formation during the late Pliocene was a major meandering river's distal low, flat floodplain, on which muds accreted that were continuously transformed into vegetated soils with Bk horizons rich in CaCO₃. The mean δ¹³C of paleosols throughout the Hadar Formation translates to an average vegetative cover across the extensive floodplain of about 30% of the C₄ grasses and 70% of unspecified C₃ plants. The character of the paleosols, such as the one at Locality 333, and their δ¹⁸O_Carbonate argue for a highly seasonal rainfall of about twice today's amount, implying that the C₃ plants were mostly sizeable trees and that the biome for Australopithecus afarensis was a grassy woodland. The amount of grasses abruptly increased in the lower Busidima Formation with its early Homo and artifacts to a more open grassy woodland of ca. 50% grasses. However, this transition in δ¹³C is not mirrored in the δ¹⁸O, which persists at a quite negative average value of −6.4‰ over the entire >2-Myr duration of both formations. This value for the carbonate means that the paleosoil water was a quite negative −4.1‰, a significant 5‰ more negative than our estimate of modern rain at Hadar. We put the negative δ¹⁸O of paleo-Hadar's rainfall into an isotopic framework of the dynamic history of climate change in sub-Saharan northern Africa. There have been two end-member climate regimes: (1) an earlier persistently pluvial Pliocene regime, with its strong summer monsoon, as registered in the Hadar Formation; and (2) the modern cyclical, mostly arid regime that began ca. 1 Myr ago, which has been punctuated by about ten cyclically predictable brief millennia-long pluvial episodes. The best known pluvial of the latter regime is the latest one, the African Humid Period (AHP), just 9.0-6.5 kyr ago, whose δ¹⁸O_Rainfall matches that for paleo-Hadar. The known climatological factors that brought on the AHP are probably the same ones that were persistently present for the Afar of the Pliocene. This dynamic rainfall history undoubtedly has influenced hominid occupation of the keystone Afar area at the gateway out of, and into, Africa.     

Mineral nutrition and heterotrophy in the water conservative holoparasite Hydnora Thunb. (Hydnoraceae)

There are large gaps in our understanding of parasite–host nutrient relationships. Our goal was to evaluate transdermal water loss, parasite–host mineral relationships, and heterotrophy in the holoparasitic genus Hydnora. We estimated in situ transdermal water loss in Hydnora and measured nutrient profiles and δ¹³C and δ¹⁵N signatures for Hydnora and hosts in southern Africa and Madagascar. For comparison we also measured δ¹³C and δ¹⁵N for aerial hemiparasites at the same sites. Transdermal water loss in Hydnora ranged from 0.14 ± 0.02 to 0.38 ± 0.04 mg cm⁻² h⁻¹ and was comparable to transpiration rates for water conservative xerophytes. Concentrations of P and K were higher in Hydnora relative to CAM hosts; other mineral concentrations were significantly lower in the parasite or were not different. δ¹³C signatures of holoparasites and hemiparasites relative to their hosts reflected host metabolism and differences in commitment to heterotrophic C gain. Holoparasite δ¹³C values were significantly enriched (by 0.55‰ ± 0.23) compared to host shoot and depleted compared to host root tissues (by −0.97‰ ± 0.12). Holoparasite δ¹³C values were not significantly different compared to the estimated whole host δ¹³C value. δ¹⁵N values for holoparasites and hemiparasites were significantly correlated with hosts. The water conservative nature of Hydnora spp. combined with parasite–host mineral nutrition profiles are suggestive of active processes of solute uptake. Stable isotope fractionation in host tissues dictated significant differences between parasite and host (shoot and root) δ¹³C signatures. The confirmation of complete heterotrophy and the lack of a confounding transpiration stream may make Hydnora a promising model organism for the examination of parasite solute uptake.     

Effects of temperature and salinity on the standard metabolic rate (SMR) of the caridean shrimp Palaemon peringueyi

The standard metabolic rate (SMR) of the caridean shrimp Palaemon peringueyi to changes in temperature (15-30 °C), salinity (0-45‰) and a combination thereof was investigated. The rate of oxygen consumption of the shrimp was determined using a YSI oxygen meter. At a constant salinity of 35‰ the respiration rate of P. peringueyi increased with an increase in temperature and ranged between 0.260 and 0.982 μl O₂ mg wwt^− 1 h^− 1. The Q₁₀ value over the temperature range 15-25 °C was estimated at 3.13. At a constant temperature of 15 °C the respiration rate of P. peringueyi also increased with an increase in salinity and ranged between 0.231 and 0.860 μl O₂ mg wwt^− 1 h^− 1. For combination experiments the absence of any significant difference in the respiration rate of P. peringueyi at the four temperatures over the salinity range 15-35‰ suggests that the shrimp is well adapted to inhabiting environments characterised by variations in salinity and temperature such as those encountered within the middle and lower reaches of permanently open estuaries with substantial freshwater inflow. On the other hand, the total mortality of the shrimp recorded at salinities < 5‰ at all four temperatures suggests that the upper distribution of the shrimp may reflect physiological constraints. Similarly, the increase in the respiration rate of the shrimp at the four temperatures at salinities > 35‰ suggests that the shrimp may experience osmotic stress in freshwater deprived permanently open and intermittently open estuaries where hypersaline conditions may develop.     

Salinities, not diets, affect strontium/calcium ratios in otoliths of Anguilla japonica

Although otolith Strontium (Sr)/calcium (Ca) ratios have been widely used to reconstruct the past salinity environmental history of anguillid eels, factors affecting the Sr/Ca ratios in otoliths are incompletely understood. Japanese Eel (Anguilla japonica) elvers (mean length 54.7 ± 2.1 mm) were collected in the estuary during their upstream migration and reared at 5 different salinities (0, 5, 15, 25, and 35 psu) and 3 types of feeding conditions (formulated feed, tubifex, and starvation) for 30 days to evaluate the effects of salinity and diets on otolith Sr/Ca ratios. Ca and Sr concentrations in the ambient water significantly increased with salinity (SAL) as [Ca] _water = 15.50SAL − 5.56, and [Sr] _water = 0.21SAL + 0.03, respectively. Sr/Ca ratios in otoliths increased with salinity (SAL) of the rearing water as [(Sr/Ca) × 1000] _otolith = 0.091SAL + 3.790. In diets, Sr/Ca ratios were 4 times higher in tubifex than in formulated feed. However, in otoliths, ANOVA indicated that Sr/Ca ratios did not differ significantly between groups fed on tubifex or formulated feed (p = 0.118). Otolith Sr/Ca ratios were negatively correlated with fish growth rates while the growth rates differed significantly among rearing conditions with different salinities and diets. Partition coefficients of the Sr/Ca ratios from ambient water to fish tissues and otoliths significantly increased with salinity. The Sr/Ca ratios of Japanese Eel otoliths thus were positively correlated with the ambient salinity and decreased with increasing fish growth rate, but was not affected by fish diet.     

Differentiation of benthic marine primary producers using stable isotopes and fatty acids: Implications to food web studies

A two-dimensional biomarker approach, using stable isotopes (δ¹³C, δ¹⁵N) and fatty acids, was used to evaluate differences both amongst and within benthic primary producer types (seagrass, fleshy red algae, calcareous red algae, brown algae, and seagrass periphyton) that are typical of the nearshore, temperate Australian region. The primary source of variance (as examined by permutational ANOVA) for all biomarkers examined was amongst primary producer types, as opposed to amongst species within type. δ¹³C showed a clear separation (Monte Carlo p < 0.05) between seagrass (range of means = −10.1 to −14.0‰) and macroalgae (−14.6 to −25.2‰), but could not differentiate amongst the algal types examined. Similarly, distinct δ¹⁵N signatures (p < 0.05) were found only for seagrass (range of means = 3.6-4.1‰) versus calcareous red algae (4.6-5.5‰), with all other types overlapping in their mean δ¹⁵N values. In contrast, multivariate analysis of fatty acid data (using Canonical Analysis of Principal coordinates; CAP) distinguished not only between seagrass and macroalgae, but also between red and brown algae (and to a limited extent between the calcareous and fleshy red algal types). The principal unsaturated fatty acids in the samples were C₂₀ polyunsaturates (found primarily in the macroalgae and periphyton), and C₁₈ mono- and polyunsaturates, with high proportions of 18:2n-6 and 18:3n-3 typical of the seagrasses. The C₁₈ monounsaturate 18:1n-7 was one of the most diagnostic compounds for the red algae examined, being present in very low amounts in seagrass and virtually absent in the brown algae. Conversely, brown algae were high in 18:4n-3, with 20:4n-3 particularly diagnostic of the kelp Ecklonia radiata. In contrast to stable isotopes, fatty acids helped distinguish different algal groups, thereby providing support that a two-dimensional approach using stable isotopes and fatty acids is likely to provide the most useful tool to distinguish primary producers in food web structure.     

Otoliths speak out: why the Pacific halibut in Puget sound are different

Pacific halibut, Hippoglossus stenolepis, is one of the most important commercial groundfish and is managed as a single coast-wide population from Alaska to northern California. Nevertheless, genetic investigations did not show success in detecting the population structure of the species. Here I report stable oxygen and carbon isotope analyses (δ¹⁸O and δ¹³C) in otoliths to discriminate the stock differences from two sample locations between the Washington coast (WC) and the northern Puget Sound (PS), and two sample years in 2007 and 2008. In general the δ¹⁸O values of halibut otoliths from WC ranged from ?0.2 to 1.8‰, higher than the PS samples from ?0.5 to 1.4‰. In contrast, the δ¹³C values from WC ranged from ?3.6 to ?1.0‰, lower than the PS samples from ?3.2 to ?1.2‰. Results from the otolith nuclei (age-0 halibut) and the 8^th (the earliest maturity age for male halibut) and edge otolith rings (the latest location where the fish lived) showed significant differences between halibut samples from PS and WC. In particular, the sample location difference (between PS and WC) in both δ¹³C and δ¹⁸O data was significant and markedly larger than the sample year difference (between 2007 and 2008). These isotopic signatures provide evidence that the PS halibut may belong to a distinct stock that is significantly different from WC halibut.     

Statistical analysis on otolith data of anadromous fishes

Stable oxygen and carbon isotope (δ¹⁸O and δ¹³C) analyses of otoliths are becoming increasingly common in fisheries science and management. However, little is known about the statistical properties of isotopic data and few attempts have been made to explore appropriate statistical methods that could be used for otolith data analysis. In this paper, we present a pilot study on δ¹⁸O and δ¹³C data from otoliths of two anadromous fish species, Atlantic salmon (Salmo salar) and Pacific sockeye salmon (Oncorhynchus nerka). The results indicated that the salmon otolith data were not normally distributed, so that linear discriminant function analysis and commonly-used statistical tests such as ANOVA and the t-test may not be appropriate. Using non-parametric k-sample nearest neighbor discriminant analysis, we were able to discriminate with high accuracy among five hatcheries for Atlantic salmon and the origins of wild and hatchery sockeye salmon. Analyses also indicated that the sample sizes required to estimate δ¹⁸O and δ¹³C means based on the different sources of variability (between group or within group) and precision levels (≤ ±5.0 %) were not large. These results and conclusions not only address the statistical considerations of isotopic data from otoliths, but also have practical importance for fisheries management as well.     

Recent and historical discharge of a large European river system - oxygen isotopic composition of river water and skeletal aragonite of Unionidae in the Rhine

Seasonal variations in the oxygen isotopic composition of Rhine River water were analyzed in detail and compared with the oxygen isotopic record from recent and historical specimens of freshwater bivalves (Unionidae). The purpose of this study was to investigate the potential use of these aragonitic bivalves as proxy recorders for varying amounts and sources of discharge, and thereby infer climate change. Seasonal variations in the river water δ¹⁸O are on the order of 1-2‰. During summer, Alpine melt-water contributes significantly to the total discharge, resulting in average values of −10 to −10.5‰, whereas the non-Alpine contribution is higher during winter, as indicated by mean δ¹⁸O values of −8.5 to −9‰. The basic pattern of the modern seasonal variation of δ¹⁸O of river water can be described by a numerical mass balance approximation of the various contributions from the Alpine and non-Alpine catchments with their average δ¹⁸O composition. The δ¹⁸O of growth increments in the prismatic shell layer of Anodonta corresponds perfectly to what is predicted by known fractionation of ¹⁸O between water and aragonite. Shell growth is restricted to water temperatures above 8-10°C, so variations in δ¹⁸O and the river water temperature are faithfully recorded by relatively large growth increments during summer. The distinctive isotopic signatures of individual flood events during summer and autumn are also recorded in the shells.     

Stable isotope analyses in otoliths of silver carp: a pilot study in identification of natal sources and stock differences

Silver carp, Hypophthalmichthys molitrix, is one of the four major fishes in China for fishing and aquaculture. The Yangtze River and associated waterbodies are important home to silver carp. In this study, we collected 135 otoliths of juvenile silver carp from the two rivers and the Poyang Lake, and analyzed for stable oxygen and carbon isotope ratios (δ¹⁸O and δ¹³C). The isotopic data and correlation of δ¹⁸O versus δ¹³C showed a clear separation and significant differences (p?<?0.001 for both δ¹³C and δ¹⁸O) between otoliths from the Ganjiang River and the Yangtze River at δ¹⁸O values around ?10‰. The Poyang Lake samples were overlapped with those of the Ganjiang River, but distinctly different from the Yangtze River samples. These isotopic signatures agree well with the previous genetic investigation results that there are marked distinctions between one population in the Ganjiang River and the other in the Yangtze River, and the Poyang Lake population belongs to the Ganjiang River rather than to the Yangtze River. Thus we concluded that stable isotope analyses of otoliths can be effectively used to study natal sources and stock differences of silver carp.     

High‐resolution isotope analysis of young alewife Alosa pseudoharengus otoliths: assessment of temporal resolution and reconstruction of habitat occupancy and thermal history

Otoliths of age 0 year alewife Alosa pseudoharengus collected in different Lake Michigan habitats were microsampled, and carbon and oxygen isotope ratios (δ¹⁸O_otolith and δ¹³C_otolith) of resulting microsamples were quantified. To assess the temporal resolution of the method, age and otolith growth rates were also estimated by counting otolith daily growth increments. Core and outer intra‐otolith samples averaged 36 and 23 days, respectively. Because of the accretionary nature of otolith growth, a habitat switch by a larva occurring between 0 and 18 days post‐hatch may not be recognized by this approach. Taking this temporal resolution into account, A. pseudoharengus habitat occupancy and thermal history in nearshore Lake Michigan, and a connecting drowned river‐mouth lake were documented. Comparisons between δ¹⁸O_otolith and δ¹³C_otolith profiles, and isotope values of Lake Michigan habitats suggested that movements by individual fish between a nearshore area of Lake Michigan proper and drowned river‐mouth lake habitats were rare. Some individuals evidently moved between habitats, and such movements occurred during different periods of ontogeny. Thermal reconstructions, based on δ¹⁸O_otolith values suggested that during early life (e.g. first month of life) young A.pseudoharengus appeared to inhabit microhabitats with temperatures greater than mean epilimnetic temperatures. This study demonstrates not only the utility of intra‐otolith geochemical analysis to describe the complexity of fish behaviour in fresh water but also identifies limitations of the present approach.     

Pre-steady state kinetic studies on the microsecond time scale of the laccase from Trametes versicolor

Laccase from Trametes versicolor reduces dioxygen to water. The enzyme is used in green chemistry applications such as the selective oxidation of alcohols in the presence of a suitable mediator (TEMPO) or in biofuel cells. We studied the catalytic mechanism of the enzyme by the stopped-flow and our newly developed rapid-mixing rapid sampling method, which has an experimental dead time of 75 ± 15 μs. Equilibrium and kinetic analyses yielded a reduction potential of 717 ± 5 mV for Type 1 copper center. EPR and low-temperature UV-Vis spectroscopy indicate that oxidation of the blue copper center and OO bond splitting occur within 100 μs, without detectable formation of a peroxide intermediate. These results indicate a rapid internal electron transfer between the various copper centers (>25.000/s) and rapid binding of O₂ (k_on > 5 × 10⁷ M⁻¹ s⁻¹). Mechanistic aspects of the catalytic cycle are shortly discussed.     

N:P ratios, δN fractionation and nutrient resorption along a nitrogen to phosphorus limitation gradient in an oligotrophic wetland complex

The vegetation N:P ratio is thought to be a diagnostic indicator of the nature of nutrient limitation in wetland vegetation. It should therefore be closely linked to other indicators of nutrient acquisition and conservation, such as nitrogen stable isotope fractionation (δ¹⁵N), nutrient resorption efficiency (RE) and resorption proficiency (RP). However, the interrelationships among these traits and the N:P ratio remain unclear. We compared tissue nutrient concentrations, N:P ratios, δ¹⁵N fractionation, RE, and RP along an N to P limitation gradient in an oligotrophic wetland valley in the South Island of New Zealand. Within the valley, the soil TN:TP ratio increased from 1.3 to 18.0 in three discrete wetlands along the gradient. In pooled data from all vegetation communities within each site, the mass-based vegetation N:P ratio correlated significantly (r² = 0.35, P < 0.01) to soil TN:TP ratios and increased from 10.2 ± 2.7 to 13.5 ± 3.6 along the N to P limitation gradient. This was accompanied by an increase in tissue δ¹⁵N enrichment from 2.05 ± 1.12‰ to 6.27 ± 1.70‰, consistent with more open N cycling and lower N demand. These trends held within all vegetation types, but were particularly strong in a Typha orientalis (C-strategist) community (soil TN:TP vs vegetation N:P correlation r² = 0.78, P < 0.001; δ¹⁵N increase from 1.81 ± 0.44‰ to 7.73 ± 1.79‰). The individual N and P concentrations and retention patterns were more species-specific and less responsive to the nutrient limitation gradient. T. orientalis maximised N resorption as N limitation increased (increasing NRE from 50.8 ± 3.3% to 71.7 ± 7.4%; reducing NRP from 0.70 ± 0.12% to 0.36 ± 0.13%) but did not alter PRE or PRP, whereas the S-strategist Schoenus pauciflorus maximised P resorption as P limitation increased (increasing PRE from 48.0 ± 5.6% to 73.5 ± 10.1%; reducing PRP from 0.053 ± 0.008% to 0.015 ± 0.004%) but did not alter NRE or NRP. These results show that the tissue N:P ratio and its associated δ¹⁵N enrichment are highly responsive indicators of the relative availability of N and P at the site and community level. However, they are not indicators of species-specific physiological requirements for N and P, or of likely responses of individual species to N or P enrichment, which are better interpreted from indicators such as RE and RP that describe nutrient retention behaviour.     

Bicarbonate use in three aquatic plants

Our study aimed to test the ability of aquatic plants to use bicarbonate when acclimated to three different bicarbonate concentrations. To this end, we performed experiments with the three species Ceratophyllum demersum, Egeria densa, Lagarosiphon major to determine photosynthetic rates under varying bicarbonate concentrations. We measured bicarbonate use efficiency, photosynthetic performance and respiration. For all species, our results revealed that photosynthetic rates were highest in replicates grown at low alkalinity. Thus, E. densa had approx. five times higher rates at low (264 ± 15 μmol O₂ g⁻¹ DW h⁻¹) than at high alkalinity (50 ± 27 μmol O₂ g⁻¹ DW h⁻¹), C. demersum had three times higher rates (336 ± 95 and 120 ± 31 μmol O₂ g⁻¹ DW h⁻¹), and L. major doubled its rates at low alkalinity (634 ± 114 and 322 ± 119 μmol O₂ g⁻¹ DW h⁻¹). Similar results were obtained for bicarbonate use efficiency by E. densa (136 ± 44 and 43 ± 10 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹) and L. major (244 ± 29 and 82 ± 24 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹). As to C. demersum, efficiency was high but unaffected by alkalinity, indicating high adaptation ability to varied alkalinities. A pH drift experiment supported these results. Overall, our results suggest that the three globally widespread worldwide species of our study adapt to low inorganic carbon availability by increasing their efficiency of bicarbonate use.