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PubMed2002

Dual-isotope SPECT using (99m)Tc-hydroxymethylene diphosphonate and (201)Tl-chloride to assess mandibular invasion by intraoral squamous cell carcinoma.

UNLABELLED: We examined mandibular invasion of intraoral squamous cell carcinoma by simultaneous bone and tumor dual-isotope SPECT using (99m)Tc-hydroxymethylene diphosphonate ((99m)Tc-HMDP) and (201)Tl-chloride ((201)Tl) and by CT. METHODS: Early and delayed simultaneous bone and tumor dual-isotope SPECT and CT were performed on 39 patients suspected of having tumor invasion of the mandible by intraoral squamous cell carcinoma. SPECT images were superimposed to project tumor location from tumor SPECT onto the osseous structures shown by bone SPECT. The CT imaging protocol consisted of 5-mm contiguous axial images. RESULTS: Histopathologic examination revealed invasion of the mandible in 13 patients and no tumor invasion in 26 patients. The results of delayed dual-isotope SPECT were exactly the same as those of early dual-isotope SPECT. On early and delayed dual-isotope SPECT, the sensitivity, specificity, and accuracy in detecting mandibular invasion by intraoral squamous cell carcinoma were 100% (13/13), 88.5% (23/26), and 92.3% (36/39), respectively. The corresponding values using CT were 45.5% (5/11), 94.7% (18/19), and 76.7% (23/30), respectively, when 9 patients were excluded because of dental artifacts. CONCLUSION: Our results suggest that superimposed early bone and tumor dual-isotope SPECT images alone may be sufficient in the diagnostic evaluation of mandibular invasion by intraoral squamous cell carcinoma.

Adult↗
PubMed1990

Use of isotope effects to characterize intermediates in mechanism-based inactivation of dopamine beta-monooxygenase by beta-chlorophenethylamine.

A mechanism for beta-chlorophenethylamine inhibition of dopamine beta-monooxygenase has been postulated in which bound alpha-aminoacetophenone is generated followed by an intramolecular redox reaction to yield a ketone-derived radical cation as the inhibitory species (Mangold, J.B., and Klinman, J.P. (1984) J. Biol. Chem. 259, 7772-7779). Based on the assumption that the ketone radical is the inhibitory intermediate, an analogous system was predicted and verified (Bossard, M.J., and Klinman, J.P. (1986) J. Biol. Chem. 261, 16421-16427). In the present study, the role of alpha-aminoacetophenone as the proposed intermediate in the inactivation by beta-chlorophenethylamine was examined in greater detail. From the interdependence of tyramine and alpha-aminoacetophenone concentrations, ketone inactivation is concluded to occur at the substrate site as opposed to potential binding at the reductant-binding site. Using beta-[2-1H]- and beta-[2-2H]chlorophenethylamine, the magnitude of the deuterium isotope effect on inactivation under second-order conditions has been found to be identical to that observed under catalytic turnover, D(kappa inact/Ki) = D(kappa cat/Km) = 6-7. By contrast, the isotope effect on inactivation under conditions of substrate and oxygen saturation, D kappa inact = 2, is 3-fold smaller than that seen on catalytic turnover, D kappa cat = 6. This reduced isotope effect for inactivation is attributed to a normal isotope effect on substrate hydroxylation followed by an inverse isotope effect on the partitioning of the enol of alpha-aminoacetophenone between oxidation to a radical cation versus protonation to regenerate ketone. These findings are unusual in that two isotopically sensitive steps are present in the inactivation pathway whereas only one is observable in turnover.

Acetophenones↗
PubMed1994

Comparative feasibility of separate or simultaneous rest thallium-201/stress technetium-99m-sestamibi dual-isotope myocardial perfusion SPECT.

UNLABELLED: Separate or simultaneous rest 201Tl/stress 99mTc-sestamibi dual-isotope SPECT are potentially efficient myocardial perfusion imaging protocols which combine the use of a high-resolution 99mTc tracer for stress perfusion assessment and 201Tl, the current single-photon agent of choice, for viability assessment. METHODS: To investigate the feasibility of dual-isotope myocardial perfusion SPECT protocols using rest 201Tl and stress sestamibi, 201Tl crosstalk into the 99mTc acquisition window (Group 1, n = 26 patients) and 99mTc crosstalk into 201Tl windows (Group 2, n = 25) were studied. For Group 1, treadmill exercise with sestamibi injection and poststress SPECT ("virgin" sestamibi images) were performed, followed by rest 201Tl injection and SPECT acquisition using dual-isotope windows (contaminated or "dual" images). For Group 2, the order was reversed: rest 201Tl SPECT (virgin 201Tl images) was performed first, followed by exercise sestamibi injection and dual-isotope SPECT. RESULTS: The contribution of 201Tl scatter to the dual sestamibi images (Group 1) was measured to be 2.9% +/- 2.1%, while 99mTc crosstalk contributed 26.7% +/- 13.0% to the dual 201Tl images (Group 2). Image quality was considered good to excellent in 92% of the sestamibi (virgin and dual) images and 88% of the virgin 201Tl SPECT, but only in 23% of the dual 201Tl studies. CONCLUSIONS: Technetium-99m crosstalk into 201Tl windows is substantial; therefore, simultaneous dual-isotope protocols, which involve assessment of 201Tl images contaminated by 99mTc, are not recommended. On the other hand, because of the small amount of 201Tl crosstalk into the 99mTc window, a separate acquisition dual-isotope approach employing the rest 201Tl (virgin)/stress sestamibi sequence is acceptable.

Aged↗
PubMed1993

Teboroxime, sestamibi and thallium-201 as markers of myocardial hypoperfusion: comparison by quantitative dual-isotope autoradiography in rabbits.

The scintigraphic assessment of myocardial hypoperfusion depends on the ability of imaging agents to delineate flow disparities. Accordingly, we compared the differential uptake of teboroxime, sestamibi and 201Tl in normal and hypoperfused myocardium using quantitative dual isotope autoradiography. Rabbits with acute coronary occlusions (n = 29) received dual isotope injections of teboroxime 201Tl or sestamibi 201Tl or single isotope tracer injections. A group of sham-operated controls (n = 15) received teboroxime and/or 201Tl. Multiple 30-mu short axis slices were collected from each heart and mounted on x-ray film along with tissue standards to independently generate separate 99mTc and 201Tl autoradiographs. Teboroxime and sestamibi produced greater normal-to-defect activity contrast than 201Tl in each dual isotope heart (range 8.4-48.9 [teboroxime] versus 2.6-12.3 [201Tl], p < 0.02 and 4.5-10.4 [sestamibi] versus 3.6-7.3 [201Tl], p < 0.03). Similar profiles were obtained in the single isotope hearts. Teboroxime produced larger autoradiographic defects than 201Tl in the dual-isotope hearts [16.0% +/- 5.6% (teboroxime) versus 12.9% +/- 5.3% (201Tl) of the LV, p < 0.02]. We conclude that the 99mTc-based perfusion agents teboroxime and, to a lesser extent, sestamibi, delineate hypoperfused myocardium more clearly than 201Tl. Teboroxime detects the largest area of hypoperfusion and may provide the most accurate assessment of myocardium at risk.

Animals↗
PubMed2000

Identifying migratory Salmo trutta using carbon and nitrogen stable isotope ratios.

Many Salmo trutta populations consist of non-anadromous (freshwater-resident) brown trout and anadromous (sea-run migratory) sea trout. Although adult brown trout and sea trout can usually be identified using differences in size and body colouration, it is not possible to easily identify eggs/alevins as the progeny of brown trout or sea trout. In this study we show that delta(13)C and delta(15)N, measured using a continuous flow isotope ratio mass spectrometer (CF-IRMS), can accurately identify fish eggs as the progeny of freshwater-resident (delta(13)C(egg) = -25.7 +/- 1.9 per thousand,delta(15)N(egg) = 9.2 +/- 1.8 per thousand) or migratory (delta(13)C(egg) = -19.9 +/- 1.1 per thousand, delta(15)N(egg) = 14. 3 +/- 1.5 per thousand) adult female Salmo trutta. Case studies show that stable isotope analysis is a more reliable technique for distinguishing anadromous adult fish than differentiation using morphological characteristics. For example, stable isotope analysis of brown trout from Loch Eck, Scotland, revealed that some individuals possessed delta(13)C and delta(15)N signatures indicative of marine feeding despite visual identification as freshwater-resident fish. It is most likely that these fish are misidentified sea trout although it possible that these fish may be brown trout that have adopted an estuarine feeding strategy to avoid interspecific competition for food within Loch Eck with salmon, powan and Arctic charr. Most stable isotope studies of fish ecology use terminal tissue sampling to provide sufficient biological material for isotopic analysis; however, our study suggests that adipose fin tissue could provide a comparable measure of delta(13)C and delta(15)N. Such a strategy would be invaluable when studying the trophic ecology or migration patterns of fish of high conservation value.

Adipose Tissue↗
PubMed2000

(18)O/(16)O and (13)C/(12)C in an ahermatypic deep-water coral Lophelia pertusa from the North Atlantic: a case of disequilibrium isotope fractionation.

The numerous stable isotope studies of scleractinian photosynthetic reef-building corals in tropical seas have demonstrated the complexity of the biological and environmental factors that give rise to their isotopic composition. Scleractinian non-photosynthetic corals of the deep cold water environment might be expected to reflect the more stable physical environment in the deep sea. However, in comparison, little is known about their isotope systematics. The present study concentrates on specimens of Lophelia pertusa from the north-eastern Atlantic, the Norwegian Shelf and Fjord. Aliquots taken from the theca represent time series and show variations in delta(13)C and delta(18)O of up to 4.7 and 2.4 per thousand, respectively. The variations seem to be related to morphological features of branching. The tendency of higher values near the tip of the polyps reported previously was detected only in some samples. The delta(18)O of the corals are in all cases more negative than the equilibrium values. There is a strong correlation between delta(13)C and delta(18)O forming linear arrays. The difference between these arrays is more pronounced in delta(13)C. The covariation of delta(13)C and delta(18)O indicates an overruling 'kinetic isotope effect'. This is in line with the behaviour of some trace elements. This isotope effect should be regarded as one manifestation of variations in partition coefficients dictated by a biological control on mineralisation (such as food or reproduction) rather than changes in the relative contribution of metabolic carbon affecting delta(13)C, and changes in temperature affecting delta(18)O.

Animals↗
PubMed1987

Simultaneous determination of pool sizes and fractional turnover rates, of deoxycholic acid, cholic acid and chenodeoxycholic acid in man by isotope dilution with 2H and 13C labels and serum sampling.

This paper describes a method for simultaneous determination of the kinetics of the three major bile acids in man using (2,2,4,4-2H4) deoxycholic acid, (24-13C) cholic acid and (24-13C) chenodeoxycholic acid. The gas chromatographic/mass spectrometric-selected ion monitoring technique used provided complete separation of deoxycholic acid, cholic acid and chenodeoxycholic acid, which permitted simultaneous measurement of isotope ratios for all three bile acids. Since measurement of all three pool sizes and fractional turnover rates in a single experiment requires different isotopic labels for deoxycholic acid and cholic acid, we investigated the in vivo stability and applicability of (2,2,4,4-2H4) deoxycholic acid as a stable isotope marker for isotope dilution studies in man. No consistent differences were observed between deoxycholic acid pool sizes and fractional turnover rates determined in serum samples after administration of (2,2,4,4-2H4) deoxycholic acid and (24-13C) deoxycholic acid. Simultaneous administration of (2,2,4,4-2H4) deoxycholic acid, (24-13C) cholic acid and (24-13C) chenodeoxycholic acid and isotope ratio measurements in serum permitted determination of pool sizes and fractional turnover rates of the three major bile acid and the 7 alpha-dehydroxylation fraction. Pool sizes, fractional turnover rates and synthesis rates (input rates) agreed well with data obtained previously with (24-13C) labels in independent studies.

Carbon Isotopes↗
PubMed1982

Pharmacokinetic equivalence of stable-isotope-labeled and unlabeled drugs. Phenobarbital in man.

Stable isotope labeling of drugs has been used in human metabolism studies because it eliminates the risk of radiation exposure accompanying use of radioactive tracers. The labeled drug can be measured by gas-chromatographic mass spectrometry (GCMS). However, if reliable pharmacokinetic data are to be obtained, one has to be certain the rate of metabolism of labeled and unlabeled drug is the same, i.e., there is no kinetic isotope effect. To evaluate this for phenobarbital (PB), three humans were infused with a 1:1 mixture of phenobarbital and 1,3-15N2-2-13C-PB. Serum was collected at regular intervals. Concentrations of labeled and unlabeled phenobarbital were determined by GCMS. Within each subject, there was no trend for concentrations of labeled phenobarbital to be higher or lower than concentrations of unlabeled phenobarbital (P greater than 0.90 for all three subjects). There was no difference in the zero time intercepts, distribution and elimination time constants and half-lives, volumes of distribution and central compartment, or clearance of the two forms of phenobarbital. Thus, no isotope effect was found. Published data on other labeled drugs and the likelihood of encountering an isotope effect based on type of isotope and its location in the molecule are discussed.

Adult↗
PubMed2004

In vivo uniform (15)N-isotope labelling of plants: using the greenhouse for structural proteomics.

Isotope labelling of proteins is important for progress in the field of structural proteomics. It enables the utilisation of the power of nuclear magnetic resonance spectroscopy (NMR) for the characterisation of the three-dimensional structures and corresponding dynamical features of proteins. The usual approach to obtain isotopically labelled protein molecules is by expressing the corresponding gene in bacterial or yeast host organisms, which grow on isotope-enriched media. This method has several drawbacks. Here, we demonstrate that it is possible to fully label a plant with (15)N-isotopes. The advantage of in vivo labelling of higher organisms is that all constituting proteins are labelled and become available as functional, post-translationally modified, correctly folded proteins. A hydroponics set-up was used to create the first example of a uniformly (15)N-labelled (> 98%) plant species, the potato plant (Solanum tuberosum L., cv. Elkana). Two plants were grown at low costs using potassium-[(15)N]-nitrate as the sole nitrogen source. At harvest time, a total of 3.6 kg of potato tubers and 1.6 kg of foliage, stolons and roots were collected, all of which were fully (15)N-labelled. Gram quantities of soluble (15)N-labelled proteins (composed mainly of the glycoprotein patatin and Kunitz-type protease inhibitors) were isolated from the tubers. NMR results on the complete proteome of potato sap and on an isolated protease inhibitor illustrate the success of the labelling procedure. The presented method of isotope labelling is easily modified to label other plants. Its envisioned impact in the field of structural proteomics of plants is discussed.

Carbon Isotopes↗
PubMed2003

Effects of temperature on isotopic enrichment in Daphnia magna: implications for aquatic food-web studies.

Laboratory experiments were conducted with Daphnia magna and Hyalella sp. grown on a single food source of known isotopic composition at a range of temperatures spanning the physiological optima for each species. Daphnia raised at 26.5 degrees C were enriched in delta(13)C and delta(15)N by 3.1 and 2.8 per thousand, respectively, relative to diet. Daphnia raised at 12.8 degrees C were enriched 1.7 and 5.0 per thousand in delta(13)C and delta(15)N, respectively. Results imply a significant negative relationship between the delta(13)C and delta(15)N of primary consumers when a temperature gradient exists. Similar responses were observed for Hyalella. Results indicate a general increase in delta(13)C enrichment and decrease in delta(15)N enrichment as temperature rises. Deviations from the commonly applied isotopic enrichment values used in aquatic ecology were attributed to changes in temperature-mediated physiological rates. Field data from a variety of sources also showed a general trend toward delta(13)C enrichment with increasing temperature in marine and lacustrine zooplankton. Multivariate regression models demonstrated that, in oligotrophic and mesotrophic lakes, zooplankton delta(13)C was related to lake-specific POM delta(13)C, lake surface temperature and latitude. Temperature-dependent isotopic separation (enrichment) between predator and prey should be taken into consideration when interpreting the significance of isotopic differences within and among aquatic organisms and ecosystems, and when assigning organisms to food-web positions on the basis of observed isotope values.

Algorithms↗
PubMed2003

Effect of preparation and preservation procedures on carbon and nitrogen stable isotope determinations from zooplankton.

A literature survey of zooplankton stable isotope studies revealed inconsistencies between authors concerning (a) fixation and (b) allowance for gut clearance of zooplankton prior to delta13C and delta15N determinations. To address whether commonly used preservation techniques induce changes in stable isotope values, fresh lake zooplankton (control) were compared with preserved (ethanol, methanol, formaldehyde, gluteraldehyde, frozen and shock frozen) material. Differences of up to 1.1 per thousand for carbon and 1.5 per thousand for nitrogen isotopic signatures were found. Even freezing, the most frequently used method identified from the literature, caused significant changes compared with the control. We advocate the use of fresh material prepared immediately whenever possible, or complementary testing of the preservative method to be used. Larger organisms are routinely eviscerated, or specific tissues are dissected, and analysed for stable isotopes to reduce errors introduced via the gut contents. Yet zooplankton gut clearance is rarely performed: the gut content assumed to be negligible relative to organism mass. Experimental determinations of relative gut mass, from both original and compiled data, range from 1-26% for different zooplankton species. Using reported isotopic values of basal resources from natural systems, we calculated that, when analysing bulk zooplankton, inclusion of the gut mass may introduce substantial errors of >3 per thousand. Thus it appears prudent to perform the simple procedure of gut clearance, especially for copepod species.

Animals↗
PubMed2006

Nitrogen isotopic composition in hair protein is different in liver cirrhotic patients.

The stable-isotopic composition of nitrogen (delta15N) or carbon (delta13C) of body tissues depends on the isotopic composition of food sources and on shifts due to isotopic fractionation during metabolism. As little is known about the effects of pathophysiological conditions we measured delta15N and delta13C values in hair and hair amino acids of patients with cirrhosis (n = 21) and compared the results with those of healthy subjects (n = 100) randomly selected from the 1987-1988 VERA German nutrition survey population. Cirrhosis was reflected in lower hair 15N abundances (6.7 vs. 9.9 per thousand delta15N; P < 0.001) whereas hair 13C abundances did not differ from healthy subjects (-19.4 vs. -19.6 per thousand 13C). Distinct patterns of delta15N and delta13C values were measured in hair amino acids. The delta15N values of phenylalanine were significantly higher in cirrhotics (P < 0.001). With the exception of isoleucine, threonine, and proline all other measured amino acids showed lower delta15N values than healthy subjects (P < 0.001). Lower hair delta15N values were associated with cirrhotic liver disease which suggests that under this condition the altered liver amino acid metabolism affects the nitrogen isotopic composition of the amino acids used for hair protein synthesis. It remains to be determined in controlled studies whether the altered nitrogen isotopic composition directly reflects the pathophysiological condition or is related to differences in dietary protein intake from plant or animal food sources.

Amino Acids↗
PubMed2002

A rapid and precise technique for measuring delta(13)C-CO(2) and delta(18)O-CO(2) ratios at ambient CO(2) concentrations for biological applications and the influence of container type and storage time on the sample isotope ratios.

A simple modification to a commercially available gas chromatograph isotope ratio mass spectrometer (GC/IRMS) allows rapid and precise determination of the stable isotopes ((13)C and (18)O) of CO(2) at ambient CO(2) concentrations. A sample loop was inserted downstream of the GC injection port and used to introduce small volumes of air samples into the GC/IRMS. This procedure does not require a cryofocusing step and significantly reduces the analysis time. The precisions for delta(13)C and delta(18)O of CO(2) at ambient concentration were +/-0.164 and +/-0.247 per thousand, respectively. This modified GC/IRMS was used to test the effects of storage on the (18)O and (13)C isotopic ratios of CO(2) at ambient concentrations in four container types. On average, the change in the (13)C-CO(2) and (18)O-CO(2) ratios of samples after one week of storage in glass vials equipped with butyl rubber stoppers (Bellco Glass Inc.) were depleted by 0.12 and by 0.20 per thousand, respectively. The (13)C ratios in aluminum canisters (Scotty II and IV, Scott Specialty Gasses) after one month of storage were depleted, on average, by 0.73 and 2.04 per thousand, respectively, while the (18)O ratios were depleted by 0.38 and 1.20 per thousand for the Scotty II and IV, respectively. After a month of storage in electropolished containers (Summa canisters, Biospheric Research Corporation), the (13)C-CO(2) and (18)O-CO(2) ratios were depleted, on average, by 0.26 and enriched by 0.30 per thousand, respectively, close to the precision of measurements. Samples were collected at a mature hardwood forest for CO(2) concentration determination and isotopic analysis. A comparison of CO(2) concentrations determined with an infrared gas analyzer and from sample voltages, determined on the GC/IRMS concurrent with the isotopic analysis, indicated that CO(2) concentrations can be determined reliably with the GC/IRMS technique. The (13)C and (18)O ratios of nighttime ecosystem-respired CO(2), determined from the intercept of Keeling plots, were -26.11 per thousand (V-PDB) and -8.81 per thousand (V-PDB-CO(2)), respectively.

Air↗
PubMed1999

Early pleistocene habitat in member 1 Olorgesailie based on paleosol stable isotopes.

Stable carbon and oxygen isotope values from soil carbonates were used to determine the vegetation context of archaeological sites and local climatic conditions represented in a approximately 0.99 Ma paleosol that is exposed laterally in the Olorgesailie basin, southern Kenya rift valley. As part of this landscape-scale project, samples of an upper Member 1 paleosol were analyzed along nearly 4 km of outcrop in three adjacent parts of the basin. Modern East African soil and plant community analogs are used to interpret the isotope ratios. The carbon isotopic composition of the paleosol carbonates indicates that the area supported a local biomass of about 75-100% C(4)plants during the period of soil formation. After averaging the data for each trench, an open C(4)grassland is represented by half of the carbon values, with wooded grassland more abundant across the paleolandscape than it is in the area now. This vegetational reconstruction is supported by the mammalian faunal assemblage, which has a high percentage of grazers. Although the relatively small sample size outside the main excavation area precludes firm characterization of vegetational diversity across the basin in upper Member 1 times, eastern and western localities in the study area may have had more woody C(3)plants than the widely sampled zone in between. Oxygen isotopes indicate that the lowland basin was slightly cooler and moister than today's semi-arid climate, with greater annual rainfall. Archaeological traces have a virtually continuous distribution across the paleolandscape, but vary in density of occurrence. With the strong evidence for C(4)grassland as the primary vegetation context across most of the study area, no habitat preference by the Acheulean toolmakers at Olorgesailie is shown in our initial comparison between carbon isotope values and stone/bone densities.

Animals↗
PubMed1999

Application of fed-batch fermentation to the preparation of isotopically labeled or selenomethionyl-labeled proteins.

An increasing demand for isotopically labeled samples for spectroscopic and crystallographic studies has led to a corresponding need for effective and efficient methods for producing these samples. The present work is based on the strategy of using an isotopically labeled compound as the growth-limiting nutrient during protein expression in Escherichia coli (DE3) strains. By using dissolved O2 and agitation rate data, the cell growth, feeding of the isotopic label, induction of protein expression, and the harvest of cells can be coordinated in a feedback controlled fermenter in a simple, easily defined manner. This approach is demonstrated for the nutrient-limited production of [U-15N]- and [U-13C, U-15N]-labeled toluene 4-monooxygenase effector protein in E. coli BL21(DE3) with isotopic abundance identical to that of the labeled precursors. For selective labeling, demonstrated with selenomethionine using methionine auxotroph E. coli B834(DE3), approximately 80-85% incorporation was obtained from methionine-dependent growth of the auxotroph followed by selenomethionine feeding and protein induction upon methionine depletion. This selective labeling is accomplished in a single culture, does not require washing or resuspension, minimizes costly incorporation of label into host cell mass prior to induction, and can be easily adapted to selective labeling with other amino acids. Moreover, cell mass yield from these experiments can be readily optimized to provide the desired level of protein for a given investigation from a single growth and purification. This combination provides an efficient, controllable option for isotopic labeling experiments.

Base Sequence↗
PubMed2003

Trophic partitioning in tropical rain forest birds: insights from stable isotope analysis.

Bird communities reach their highest taxonomic and trophic diversity in tropical rain forest, but the use of different foraging strategies to meet food requirements in such competitive environments is poorly understood. Conventional dietary analyses are poorly suited to investigate dietary patterns in complex systems. We used stable carbon ((13)C/(12)C) and nitrogen ((15)N/(14)N) isotope analysis of whole blood to examine avian trophic patterns and sources of diet in the tropical rain forest of Los Tuxtlas, Veracruz, Mexico. We used stable nitrogen isotope analysis to delineate trophic levels, and stable carbon isotope analysis to distinguish the relative contribution of C-3 and CAM/C-4 ultimate sources of proteins to diets. There was large inter- and intraspecific variation in whole blood delta(13)C and delta(15)N values in 23 species of birds. Stable nitrogen isotope analysis separated birds into several trophic levels, including species that obtained their dietary protein mostly from plants, insects or a combination of both food sources. Stable carbon isotope analysis showed that most birds fed on C3-based foods but Stub-tailed Spadebills (Platyrinchus cancrominus) included C-3- and C-4/CAM-specialist individuals. Our analyses provided insights into the nutritional contribution of plant and animal sources of protein and distinguish their photosynthetic origin over relatively long average time periods.

Animals↗
PubMed2006

Changes in stable isotopic signatures of soil nitrogen and carbon during 40 years of forest development.

Understanding what governs patterns of soil delta15N and delta13C is limited by the absence of these data assembled throughout the development of individual ecosystems. These patterns are important because stable isotopes of soil organic N and C are integrative indicators of biogeochemical processing of soil organic matter. We examined delta15N of soil organic matter (delta15NSOM) and delta13CSOM of archived soil samples across four decades from four depths of an aggrading forest in southeastern USA. The site supports an old-field pine forest in which the N cycle is affected by former agricultural fertilization, massive accumulation of soil N by aggrading trees over four decades, and small to insignificant fluxes of N via NH3 volatilization, nitrification, and denitrification. We examine isotopic data and the N and C dynamics of this ecosystem to evaluate mechanisms driving isotopic shifts over time. With forest development, delta13CSOM became depth-dependent. This trend resulted from a decline of approximately 2 per thousand in the surficial 15 cm of mineral soil to -26.0 per thousand, due to organic matter inputs from forest vegetation. Deeper layers exhibited relatively little trend in delta13CSOM with time. In contrast, delta15NSOM was most dynamic in deeper layers. During the four decades of forest development, the deepest layer (35-60 cm) reached a maximum delta15N value of 9.1 per thousand, increasing by 7.6 per thousand. The transfer of > 800 kg ha(-1) of soil organic N into aggrading vegetation and the forest floor and the apparent large proportion of ectomycorrhizal (ECM) fungi in these soils suggest that fractionation via microbial transformations must be the major process changing delta15N in these soils. Accretion of isotopically enriched compounds derived from microbial cells (i.e., ECM fungi) likely promote isotopic enrichment of soils over time. The work indicates the rapid rate at which ecosystem development can impart delta15NSOM and delta13CSOM signatures associated with undisturbed soil profiles.

Carbon Isotopes↗
PubMed2006

Isotopic enrichment in a phloem-feeding insect: influences of nutrient and water availability.

The isotopic enrichment between an animal and its diet can vary among and within living systems, but the sources of variation are not yet fully understood. Some studies have found that diet quality or an animal's nutritional status can influence the degree of trophic enrichment, while others have dismissed nutrition as a contributing factor. We evaluated the effects of nutrient and water availability on carbon and nitrogen isotopic enrichment in a specialized plant-herbivore system. Aphids are largely sedentary and rely exclusively on nitrogen-poor phloem sap of their host for nutrition. We grew potato aphids [Macrosiphum euphorbiae (Aphididae)] on an accepted host, pumpkin [Cucurbita pepo L. (Cucurbitaceae)], in a glasshouse environment. Twelve pumpkin plants growing under high- and low-watering regimes were inoculated at 4 weeks of age with aphids. During the course of the experiment we collected leaves, phloem sap, aphids and honeydew (i.e., aphid exudates). We found no trophic enrichment between aphids and their phloem sap diet, but significant carbon enrichment of honeydew relative to aphids (2.5 per thousand) and phloem sap (2.1 per thousand). Honeydew was also enriched in nitrogen compared to the phloem sap (1.2 per thousand). Watering treatment had a substantial impact on trophic enrichment. Correlations among tissues, an indication of uniform trophic enrichment among samples, were significant only for the carbon isotopic composition, and then only for plants and aphids grown in the low-water treatment. Diet quality also influenced the degree of isotopic enrichment; trophic enrichment for both carbon and nitrogen isotopic composition increased as diet quality (C/N) declined. We conclude that the degree of trophic enrichment is variable due, in part, to diet quality, but that the scale of variation is small.

Analysis of Variance↗