Sources and pathways of carbon and nitrogen of macrophytes and sediments using stable isotopes in Al-Kharrar Lagoon, eastern Red Sea coast, Saudi Arabia.

Elemental ratios (δ13C, δ15N and C/N) and carbon and nitrogen concentrations in macrophytes, sediments and sponges of the hypersaline Al-Kharrar Lagoon (KL), central eastern Red Sea coast, were measured to distinguish their sources, pathways and see how they have been influenced by biogeochemical processes and terrestrial inputs. The mangroves and halophytes showed the most depleted δ13C values of -27.07±0.2 ‰ and -28.34±0.4 ‰, respectively, indicating their preferential 12C uptake, similar to C3-photosynthetic plants, except for the halophytes Atriplex sp. and Suaeda vermiculata which showed δ13C of -14.31±0.6 ‰, similar to C4-plants. Macroalgae were divided into A and B groups based on their δ13C values. The δ13C of macroalgae A averaged -15.41±0.4 ‰, whereas macroalgae B and seagrasses showed values of -7.41±0.8 ‰ and -7.98 ‰, suggesting uptake of HCO3- as a source for CO2 during photosynthesis. The δ13C of sponges was -10.7±0.3 ‰, suggesting that macroalgae and seagrasses are their main favoured diets. Substrates of all these taxa showed δ13C of -15.52±0.8 ‰, suggesting the KL is at present a macroalgae-dominated lagoon. The δ15N in taxa/sediments averaged 1.68 ‰, suggesting that atmospheric N2-fixation is the main source of nitrogen in/around the lagoon. The heaviest δ15N (10.58 ‰) in halophytes growing in algal mats and sabkha is possibly due to denitrification and ammonia evaporation. The macrophytes in the KL showed high C %, N %, and C/N ratios, but this is not indicated in their substrates due possibly to a rapid turnover of dense, hypersaline waters carrying most of the detached organic materials out into the Red Sea. The δ13C allowed separation of subaerial from aquatic macrophytes, a proxy that could be used when interpreting paleo-sea level or paleoclimatic changes from the coastal marine sediments.

Amino acid-specific isotopes reveal changing five-dimensional niche segregation in Pacific seabirds over 50 years.

Hutchison's niche theory suggests that coexisting competing species occupy non-overlapping hypervolumes, which are theoretical spaces encompassing more than three dimensions, within an n-dimensional space. The analysis of multiple stable isotopes can be used to test these ideas where each isotope can be considered a dimension of niche space. These hypervolumes may change over time in response to variation in behaviour or habitat, within or among species, consequently changing the niche space itself. Here, we use isotopic values of carbon and nitrogen of ten amino acids, as well as sulphur isotopic values, to produce multi-isotope models to examine niche segregation among an assemblage of five coexisting seabird species (ancient murrelet Synthliboramphus antiquus, double-crested cormorant Phalacrocorax auritus, Leach's storm-petrel Oceanodrama leucorhoa, rhinoceros auklet Cerorhinca monocerata, pelagic cormorant Phalacrocorax pelagicus) that inhabit coastal British Columbia. When only one or two isotope dimensions were considered, the five species overlapped considerably, but segregation increased in more dimensions, but often in complex ways. Thus, each of the five species occupied their own isotopic hypervolume (niche), but that became apparent only when factoring the increased information from sulphur and amino acid specific isotope values, rather than just relying on proxies of δ15N and δ13C alone. For cormorants, there was reduction of niche size for both species consistent with a decline in their dominant prey, Pacific herring Clupea pallasii, from 1970 to 2006. Consistent with niche theory, cormorant species showed segregation across time, with the double-crested demonstrating a marked change in diet in response to prey shifts in a higher dimensional space. In brief, incorporating multiple isotopes (sulfur, PC1 of δ15N [baselines], PC2 of δ15N [trophic position], PC1 and PC2 of δ13C) metrics allowed us to infer changes and differences in food web topology that were not apparent from classic carbon-nitrogen biplots.

Changes in urinary stable nitrogen isotope ratios during controlled short-term energy deficit: a proof-of-principle analysis.

PURPOSE: Stable isotope ratios of nitrogen (δ15N) have previously been shown to increase in human hair during periods of catabolism. The goal of this study was to assess changes in δ15N in urinary urea (δ15Nurea) and Δ15N during a short-term controlled energy deficit. METHODS: We analyzed samples from 6 recreationally active men (25 ± 1 years, BMI: 23.5 ± 0.6 kg/m2) who participated in a repeated measures cross-over study involving 4 days of energy deficit (ED, ~ 15 kcal/kg FFM) without and with exercise (ED-EX, ED + EX) and control conditions in energy balance (CON-EX, CON + EX). δ15Nurea was analyzed from urine samples, and Δ15N was calculated as δ15Nurea-δ15Ndiet, with δ15Ndiet obtained from diet prescriptions. RESULTS: δ15Nurea was significantly elevated in ED-EX (4.4 ± 0.2‰) when compared to CON-EX (3.7 ± 0.1‰; p = 0.026) and CON + EX (3.34 ± 0.13‰, p = 0.001). As a consequence, Δ15N was positive in ED-EX (0.2 ± 0.2‰) and remained negative in ED + EX (- 0.6 ± 0.5‰), CON-EX (- 1.0 ± 0.2) and CON + EX (- 1.1 ± 0.2). Differences in Δ15N were significant between ED-EX and CON-EX (p = 0.005) and ED-EX and CON + EX (p = 0.006). CONCLUSION: Our results suggest that δ15Nurea and subsequently Δ15N are responsive to a short-term energy deficit, likely due to increased amino acid oxidation to meet energy demands and preferable elimination of 14N.

Physiological Influence of Fe and Cu Availability on Nitrogen Isotope Fractionation during Ammonia Oxidation.

Microbially mediated cycling processes play central roles in regulating the speciation and availability of nitrogen, a vital nutrient with wide implications for agriculture, water quality, wastewater treatment, ecosystem health, and climate change. Ammonia oxidation, the first and rate-limiting step of nitrification, is carried out by bacteria (AOB) and archaea (AOA) that require the trace metal micronutrients copper (Cu) and iron (Fe) for growth and metabolic catalysis. While stable isotope analyses for constraining nitrogen cycling are commonly used, it is unclear whether metal availability may modulate expression of stable isotope fractionation during ammonia oxidation, by varying growth or through regulation of metabolic metalloenzymes. We present the first study examining the influence of Fe and Cu availability on the kinetic nitrogen isotope effect in ammonia oxidation (15εAO). We report a general independence of 15εAO from the growth rate in AOB, except at a low temperature (10 °C). With AOA Nitrosopumilus maritimus SCM1, however, 15εAO decreases nonlinearly at lower oxidation rates. We examine assumptions involved in the interpretation of 15εAO values and suggest these dynamics may arise from physiological constraints that push the system toward isotopic equilibrium. These results suggest important links between isotope fractionation and environmental constraints on physiology in these key N cycling microorganisms.

Interspecific, ontogenetic and temporal variations in stable isotopes of small tuna species in the northeast Atlantic Ocean.

In order to study the trophic level of small tuna species and their contribution to the carbon flow in pelagic food webs, an analysis of carbon and nitrogen stable isotopes was carried out. The investigation was focused on four small tuna species (Auxis rochei, Auxis thazard, Euthynnus alletteratus and Sarda sarda) commonly harvested in the northeast Atlantic Ocean. The isotope analysis showed how the results for S. sarda are different from the rest of the species analysed, with a higher trophic level, similar to other major tuna species. The greatest niche overlap in δ13C and δ15N occurs among A. rochei, A. thazard and E. alletteratus. Auxis rochei and E. alletteratus showed a size-dependent variability in δ15N, and in δ13C for S. sarda. The small tuna S. sarda exhibits the highest migration rates among various geographical areas in comparison to other small pelagic tunas, and the seasonal variability of isotope values in the area studied can be attributed to the incorporation of larger individuals with a higher lipid content. The results of this work provide new information on the ecological role played by small tuna in food webs, which is more complex and varied than currently thought. This knowledge is essential for a more effective management of fisheries.

Trophic Transfer of Halogenated Organic Pollutants in a Wetland Food Web: Insights from Compound-Specific Nitrogen Isotope of Amino Acids and Food Source Analysis.

A trophic position (TP) model (TPmix model) that simultaneously considered trophic discrimination factor and ßGlu/Phe variations was developed in this study and was first applied to investigate the trophic transfer of halogenated organic pollutants (HOPs) in wetland food webs. The TPmix model characterized the structure of the wetland food web more accurately and significantly improved the reliability of TMF compared to the TPbulk, TPAAs, and TPsimmr models, which were calculated based on the methods of stable nitrogen isotope analysis of bulk, traditional AAs-N-CSIA, and weighted ßGlu/Phe, respectively. Food source analysis revealed three interlocking food webs (kingfisher, crab, and frogs) in this wetland. The highest HOP biomagnification capacities (TMFmix) were found in the kingfisher food web (0.24-82.0), followed by the frog (0.08-34.0) and crab (0.56-11.7) food webs. The parabolic trends of TMFmix across combinations of log KOW in the frog food web were distinct from those of aquatic food webs (kingfisher and crab), which may be related to differences in food web composition and HOP bioaccumulation behaviors between aquatic and terrestrial organisms. This study provides a new tool to accurately study the trophic transfer of contaminants in wetlands and terrestrial food webs with diverse species and complex feeding relationships.

Comparison of markers to estimate microbial adherence and fibrolytic activity in forages after in vitro incubation.

The aims of this study were to evaluate the relationship between attached microbial markers and degradability of forage samples incubated in vitro and to compare these microbial markers. In Trial 1, the content of 15N, purines, and phosphorus (P) as well as xylanase activity in residue of different forage species were measured after 24 h of incubation in a conventional in vitro system at pH 6.8. Trial 2 used the same procedures as those of Trial 1 except that forage samples were incubated in media with different initial pH (5.5, 6.0, 6.5, or 7.0). There was no correlation (P > 0.10) between forage degradability and either microbial marker in Trial 1. Degradability of both, bermuda and ryegrass, and the content of all markers in the incubation residues was positively affected by increased pH (P < 0.05). The content of 15N in residues was linearly related to xylanase activity (P < 0.05) but not with P or purines content. In conclusion, the nutritional potential of different forage species may not to be compared, based on the content of microbial markers in the incubation residues. In other way, within a forage species, the in vitro degradation was directly associated to either marker. However, P presents analytical advantage over other markers.

Using stable isotope (δ13C, δ15N) values from feces and breath to infer shorebird diets.

The use of stable isotopes of carbon (δ13C) and nitrogen (δ15N) from feces and breath offers potential as non-destructive tools to assess diets and nutrition. How stable isotope values derived from breath and feces compare with those from commonly used tissues, such as blood fractions and liver, remains uncertain, including understanding the metabolic routing of dietary nutrients. Here, we measured δ13C and δ15N from feces and δ13C of breath from captive Red-necked Stints (Calidris ruficollis) and 26 species of wild-caught migratory shorebirds (n = 259 individuals) and compared them against isotopic values from blood and feathers. For captive birds fed either cereal- or fish-based diets, differences in δ13C between feces and lipid-free diet were small, - 0.2 ± 0.5‰ and 0.1 ± 0.3‰, respectively, and differences in δ15N, - 0.7 ± 0.5‰ and - 0.5 ± 0.5‰, respectively. Hence, δ13C and δ15N values from feces can serve as proxies for ingested proteinaceous tissues and non-soluble carbohydrates because isotopic discrimination can be considered negligible. Stable isotope values in plasma and feces were strongly correlated in wild-caught shorebirds, indicating feces can be used to infer assimilated macronutrients. Breath δ13C was 1.6 ± 0.8‰ to 5.6 ± 1.2‰ lower than bulk food sources, and breath C derived from lipids was estimated at 47.5% (cereal) to 96.1% (fish), likely underlining the importance of dietary lipids for metabolism. The findings validate the use of stable isotope values of feces and breath in isotopic assays to better understand the dietary needs of shorebirds.

Insights into amino acid fractionation and incorporation by compound-specific carbon isotope analysis of three-spined sticklebacks.

Interpretation of stable isotope data is of upmost importance in ecology to build sound models for the study of animal diets, migration patterns and physiology. However, our understanding of stable isotope fractionation and incorporation into consumer tissues is still limited. We therefore measured the δ13C values of individual amino acids over time from muscle and liver tissue of three-spined sticklebacks (Gasterosteus aculeatus) on a high protein diet. The δ13C values of amino acids in the liver quickly responded to small shifts of under ± 2.0‰ in dietary stable isotope compositions on 30-day intervals. We found on average no trophic fractionation in pooled essential (muscle, liver) and non-essential (muscle) amino acids. Negative Δδ13C values of - 0.7 ± 1.3‰ were observed for pooled non-essential (liver) amino acids and might indicate biosynthesis from small amounts of dietary lipids. Trophic fractionation of individual amino acids is reported and discussed, including unusual Δδ13C values of over + 4.9 ± 1.4‰ for histidine. Arginine and lysine showed the lowest trophic fractionation on individual sampling days and might be useful proxies for dietary sources on short time scales. We suggest further investigations using isotopically enriched materials to facilitate the correct interpretation of ecological field data.

Free amino acids, carbon and nitrogen isotopic compositions responses to cadmium stress in two castor (Ricinus communis L.) species.

Cadmium (Cd) toxicity induce various disturbances in metabolic processes and impair plant establishment. The composition of carbon and nitrogen stable isotopes (δ13C and δ15N) and free amino acids (FAAs) can reflect the response of plants to environmental stress. In the present study, a solution culture experiment was carried out, and the secretion characteristics of FAAs as well as δ13C and δ15N were evaluated as indicative of the functional performance of two castor species (Zibo-3 and Zibo-9) under various Cd concentrations stress (0, 1, 2, and 5 mg L-1). The results indicated that: 1) The treatment of the plants with 5 mg L-1 of a Cd solution resulted in a significant decline of biomasses by 22.4% and 11.6% in Zibo-3 and Zibo-9, respectively, relative to controls; additionally, the accumulation levels for Cd in Zibo-9 were higher than those in Zibo-3, thus Zibo-9 showed higher tolerance and enrichment ability to Cd. 2) The exposure of castor to Cd treatments results in significant modifications in individual FAAs, suggesting a differential sensitivity of each biosynthetic pathway to this stress; however, a positive correlation was found between the accumulation of total FAAs and Cd treatment dosages; higher proportion of asparagine and glutamate in total amino acids for Zibo-9, and abundant secretion of arginine in Cd treated Zibo-9 may be associated with the higher Cd-tolerance and Cd-accumulation in Zibo-9. 3) Cd stress increased leaf δ13C and δ15N values regardless of the castor species; δ13C and δ15N could be used as monitoring tools for heavy metal stress in plants.

Timing is everything - obtaining accurate measures of plant uptake of amino acids.

Plants are known to have the capacity to take up and utilise amino acids for growth. The significance of this uptake, however, remains elusive, partly due to methodological challenges and biological implications associated with acquiring and interpreting data. This study compared bulk stable isotope analysis and compound-specific liquid chromatography-mass spectrometry, two established methods for determining amino acid uptake. Root amino acid uptake was assayed using U-13 C5 -15 N2 -l-glutamine and axenically grown Arabidopsis thaliana. After 15-120 min of exposure, the content of intact glutamine measured in the roots was constant, whilst the 15 N and 13 C content increased over time, resulting in very different estimated uptake rates. The 13 C : 15 N ratio in roots declined with time, suggesting a loss of glutamine carbon of up to 15% within 120 min. The results presented indicate that, regardless of method used, time is a crucial factor when determining plant amino acid uptake. Due to post-uptake metabolism, compound-specific methods should primarily be used in experiments with a time frame of minutes rather than hours or days. Post-uptake metabolism in plants may account for significant loss of carbon, suggesting that it is not just pre-uptake metabolism by microbes that accounts for the 15 N-13 C mismatch reported in ecological studies, but also post-uptake metabolism in the plant.

The True Amino Acid Digestibility of 15N-Labelled Sunflower Biscuits Determined with Ileal Balance and Dual Isotope Methods in Healthy Humans.

BACKGROUND: Sunflower is a promising protein source but data on amino acid (AA) digestibility are lacking in humans. Classically, the determination of AA digestibility requires ileal digesta sampling. The dual isotope method is minimally invasive but has not been compared to the conventional approach. OBJECTIVES: This study aimed to determine the true ileal digestibility of sunflower AAs in healthy volunteers who ate biscuits containing 15nitrogen (N) protein isolate, in comparison with the dual isotope method. METHODS: Twelve healthy volunteers (men and women; 40.4 ± 10.5 years old; BMI, 23.7 ± 2.9 kg/m2) were equipped with a naso-ileal tube. For 4 hours, they consumed 9 repeated meals comprising 15N-sunflower protein biscuits together with 13carbon (C)-AAs, carried either in chocolate (SUN + Ch; n = 7) or apple puree (SUN + P; n = 5). Ileal digesta and blood were sampled throughout 8 hours after ingestion of the first meal. The 15N and 13C AA enrichments were measured in digesta to determine ileal digestibility directly and in plasma to determine lysine and threonine digestibility using the dual isotope method. Differences between methods and between vector groups were analyzed using paired and unpaired t-tests, respectively. RESULTS: The ileal digestibility of sunflower indispensable AAs (IAA) was 89% ± 5.3%, with threonine and lysine having the lowest digestibility. In the SUN + Ch meal, IAA digestibility was 3% below that of SUN + P (P < 0.05). The mean free 13C-AA ileal digestibility was 98.1% ± 0.9%. No matter which matrix was used to carry 13C-AAs, plasma 15N and 13C-AA kinetics displayed a 1-hour offset. Digestibility obtained with the dual isotope method (70.4% ± 6.0% for threonine and 75.9% ± 22.3% for lysine) was below the target values. CONCLUSIONS: The ileal digestibility of IAAs from a sunflower isolate incorporated in a biscuit was close to 90% in healthy adults. Under our experimental conditions, the dual isotope method provided lower values than the usual method. Further protocol developments are needed to validate the equivalence between both methods. This trial was registered at clinicaltrials.gov as NCT04024605.

Nitrogen Kinetic Isotope Effects of Nitrification by the Complete Ammonia Oxidizer Nitrospira inopinata.

Analysis of nitrogen isotope fractionation effects is useful for tracing biogeochemical nitrogen cycle processes. Nitrification can cause large nitrogen isotope effects through the enzymatic oxidation of ammonia (NH3) via nitrite (NO2-) to nitrate (NO3-) (15εNH4+→NO2- and 15εNO2-→NO3-). The isotope effects of ammonia-oxidizing bacteria (AOB) and archaea (AOA) and of nitrite-oxidizing bacteria (NOB) have been analyzed previously. Here, we studied the nitrogen isotope effects of the complete ammonia oxidizer (comammox) Nitrospira inopinata that oxidizes NH3 to NO3-. At high ammonium (NH4+) availability (1 mM) and pH between 6.5 and 8.5, its 15εNH4+→NO2- ranged from -33.1 to -27.1‰ based on substrate consumption (residual substrate isotopic composition) and -35.5 to -31.2‰ based on product formation (cumulative product isotopic composition), while the 15εNO2-→NO3- ranged from 6.5 to 11.1‰ based on substrate consumption. These values resemble isotope effects of AOB and AOA and of NOB in the genus Nitrospira, suggesting the absence of fundamental mechanistic differences between key enzymes for ammonia and nitrite oxidation in comammox and canonical nitrifiers. However, ambient pH and initial NH4+ concentrations influenced the isotope effects in N. inopinata. The 15εNH4+→NO2- based on product formation was smaller at pH 6.5 (-31.2‰) compared to pH 7.5 (-35.5‰) and pH 8.5 (-34.9‰), while 15εNO2-→NO3- was smaller at pH 8.5 (6.5‰) compared to pH 7.5 (8.8‰) and pH 6.5 (11.1‰). Isotopic fractionation via 15εNH4+→NO2- and 15εNO2-→NO3- was smaller at 0.1 mM NH4+ compared to 0.5 to 1.0 mM NH4+. Environmental factors, such as pH and NH4+ availability, therefore need to be considered when using isotope effects in 15N isotope fractionation models of nitrification. IMPORTANCE Nitrification is an important nitrogen cycle process in terrestrial and aquatic environments. The discovery of comammox has changed the view that canonical AOA, AOB, and NOB are the only chemolithoautotrophic organisms catalyzing nitrification. However, the contribution of comammox to nitrification in environmental and technical systems is far from being completely understood. This study revealed that, despite a phylogenetically distinct enzymatic repertoire for ammonia oxidation, nitrogen isotope effects of 15εNH4+→NO2- and 15εNO2-→NO3- in comammox do not differ significantly from those of canonical nitrifiers. Thus, nitrogen isotope effects are not suitable indicators to decipher the contribution of comammox to nitrification in environmental samples. Moreover, this is the first systematic study showing that the ambient pH and NH4+ concentration influence the isotope effects of nitrifiers. Hence, these key parameters should be considered in comparative analyses of isotope effects of nitrifiers across different growth conditions and environmental samples.

Impact of intensifying nitrogen limitation on ocean net primary production is fingerprinted by nitrogen isotopes.

The open ocean nitrogen cycle is being altered by increases in anthropogenic atmospheric nitrogen deposition and climate change. How the nitrogen cycle responds will determine long-term trends in net primary production (NPP) in the nitrogen-limited low latitude ocean, but is poorly constrained by uncertainty in how the source-sink balance will evolve. Here we show that intensifying nitrogen limitation of phytoplankton, associated with near-term reductions in NPP, causes detectable declines in nitrogen isotopes (δ15N) and constitutes the primary perturbation of the 21st century nitrogen cycle. Model experiments show that ~75% of the low latitude twilight zone develops anomalously low δ15N by 2060, predominantly due to the effects of climate change that alter ocean circulation, with implications for the nitrogen source-sink balance. Our results highlight that δ15N changes in the low latitude twilight zone may provide a useful constraint on emerging changes to nitrogen limitation and NPP over the 21st century.

Diet-tissue discrimination and turnover of δ13 C and δ15 N in muscle tissue of a penaeid prawn.

RATIONALE: Stable isotopes are used to study trophic and movement ecology in aquatic systems, as they provide spatially distinct, time-integrated signatures of diet. Stable isotope ecology has been used to quantify species-habitat relationships in many important fisheries species (e.g., penaeid prawns), with approaches that typically assume constant values for diet-tissue discrimination and diet-tissue steady state, but these can be highly variable. Here we provide the first report of these processes in Metapenaeus macleayi (eastern school prawn). METHODS: Here we explicitly measure and model carbon (δ13 C) and nitrogen (δ15 N) diet-tissue discrimination and turnover in eastern school prawn muscle tissue as a function of experimental time following a change in diet to an isotopically distinct food source. RESULTS: Diet-tissue discrimination factors were 5 and 0.6‰ for δ13 C and δ15 N, respectively. Prawn muscle tissue reached an approximate steady state after approximately 50 and 30 days for δ13 C and δ15 N. Half-lives indicated faster turnover of δ15 N (~8 days) than δ13 C (~14 days). CONCLUSIONS: Our diet-tissue discrimination factors deviate from 'typical' values with larger values for carbon than nitrogen isotopes, but are generally similar to those measured in other crustaceans. Similarly, our estimates of isotopic turnover align with those in other penaeid species. These findings confirm muscle tissue as a reliable indicator of long-term diet and movement patterns in eastern school prawn.

Aggregation Condition-Structure Relationship of Mouse Prion Protein Fibrils.

Prion diseases are associated with conformational conversion of cellular prion protein into a misfolded pathogenic form, which resembles many properties of amyloid fibrils. The same prion protein sequence can misfold into different conformations, which are responsible for variations in prion disease phenotypes (prion strains). In this work, we use atomic force microscopy, FTIR spectroscopy and magic-angle spinning NMR to devise structural models of mouse prion protein fibrils prepared in three different denaturing conditions. We find that the fibril core region as well as the structure of its N- and C-terminal parts is almost identical between the three fibrils. In contrast, the central part differs in length of ß-strands and the arrangement of charged residues. We propose that the denaturant ionic strength plays a major role in determining the structure of fibrils obtained in a particular condition by stabilizing fibril core interior-facing glutamic acid residues.

An arithmetic correction for the effect of lipid on carbon stable isotope ratios in muscle and digestive glands of the American lobster (Homarus americanus).

RATIONALE: Lipid correction models use elemental carbon-to-nitrogen ratios to estimate the effect of lipids on δ13 C values and provide a fast and inexpensive alternative to chemically removing lipids. However, the performance of these models varies, especially in whole-body invertebrate samples. The generation of tissue-specific lipid correction models for American lobsters, both an ecologically and an economically important species in eastern North America, will aid ecological research of this species and our understanding of the function of these models in invertebrates. METHOD: We determined the δ13 C and δ15 N values before and after lipid extraction in muscle and digestive glands of juvenile and adult lobster. We assessed the performance of four commonly used models (nonlinear, linear, natural logarithm (LN) and generalized linear model (GLM)) at estimating lipid-free δ13 C values based on the non-lipid-extracted δ13 C values and elemental C:N ratios. The accuracy of model predictions was tested using paired t-tests, and the performance of the different models was compared using the Akaike information criterion score. RESULTS: Lipid correction models accurately estimated post-lipid-extraction δ13 C values in both tissues. The nonlinear model was the least accurate for both tissues. In muscle, the three other models performed well, and in digestive glands, the LN model provided the most accurate estimates throughout the range of C:N values. In both tissues, the GLM estimates were not independent of the post-lipid-extraction δ13 C values, thus reducing their transferability to other datasets. CONCLUSIONS: Whereas previous work found that whole-body models poorly estimated the effect of lipids in invertebrates, we show that tissue-specific lipid correction models can generate accurate and precise estimates of lipid-free δ13 C values in lobster. We suggest that the tissue-specific logarithmic models presented here are the preferred models for accounting for the effect of lipid on lobster isotope ratios.

A new carnivorous plant lineage (Triantha) with a unique sticky-inflorescence trap.

Carnivorous plants consume animals for mineral nutrients that enhance growth and reproduction in nutrient-poor environments. Here, we report that Triantha occidentalis (Tofieldiaceae) represents a previously overlooked carnivorous lineage that captures insects on sticky inflorescences. Field experiments, isotopic data, and mixing models demonstrate significant N transfer from prey to Triantha, with an estimated 64% of leaf N obtained from prey capture in previous years, comparable to levels inferred for the cooccurring round-leaved sundew, a recognized carnivore. N obtained via carnivory is exported from the inflorescence and developing fruits and may ultimately be transferred to next year's leaves. Glandular hairs on flowering stems secrete phosphatase, as seen in all carnivorous plants that directly digest prey. Triantha is unique among carnivorous plants in capturing prey solely with sticky traps adjacent to its flowers, contrary to theory. However, its glandular hairs capture only small insects, unlike the large bees and butterflies that act as pollinators, which may minimize the conflict between carnivory and pollination.

Distinct nitrogen isotopic compositions of healthy and cancerous tissue in mice brain and head&neck micro-biopsies.

BACKGROUND: Cancerous cells can recycle metabolic ammonium for their growth. As this ammonium has a low nitrogen isotope ratio (15N/14N), its recycling may cause cancer tissue to have lower 15N/14N than surrounding healthy tissue. We investigated whether, within a given tissue type in individual mice, tumoral and healthy tissues could be distinguished based on their 15N/14N. METHODS: Micro-biopsies of murine tumors and adjacent tissues were analyzed for 15N/14N using novel high-sensitivity methods. Isotopic analysis was pursued in Nude and C57BL/6 mice models with mature orthotopic brain and head&neck tumors generated by implantation of H454 and MEERL95 murine cells, respectively. RESULTS: In the 7 mice analyzed, the brain tumors had distinctly lower 15N/14N than healthy neural tissue. In the 5 mice with head&neck tumors, the difference was smaller and more variable. This was at least partly due to infiltration of healthy head&neck tissue by tumor cells. However, it may also indicate that the 15N/14N difference between tumoral and healthy tissue depends on the nitrogen metabolism of the healthy organ in question. CONCLUSIONS: The findings, coupled with the high sensitivity of the 15N/14N measurement method used here, suggest a new approach for micro-biopsy-based diagnosis of malignancy as well as an avenue for investigation of cancer metabolism.

Testing of how and why the Terpios hoshinota sponge kills stony corals.

An encrusting sponge, Terpios hoshinota, has the potential to infect all species of stony corals in shallow reefs and killing them. It caused a decline in coral coverage in two south-eastern islands of Taiwan. We proposed two hypotheses to examine how the sponges kill the corals, namely, light blocking and toxins, and tested by in-situ experiments. The results revealed that both light blocking, sponge toxins, and particularly the combination of both factors were effective at inducing tissue damage in stony corals over a short period. Second, to answer why the sponges killed the corals, we tested two hypotheses, namely, gaining nutrients versus gaining substrates for the sponge. By analyzing the stable isotopes 13C and 15N, as well as exploiting an enrichment experiment, it was possible to determine that only approximately 9.5% of the carbon and 16.9% of the nitrogen in the newly grown sponge tissues originated from the enriched corals underneath. The analysis also revealed that the control corals without isotope enrichment had higher δ13C and δ15N than the control sponges, which was an additional indication that T. hoshinota did not rely heavily on corals for nutrients. Therefore, our results support the hypothesis that the encrusting sponge did not kill corals for food or nutrients, but rather for the substrate.