Differences In δ13C Values Research Articles

Spatial ecology of moose in Sweden: Combined Sr-O-C isotope analyses of bone and antler.

The study of spatial (paleo)ecology in mammals is critical to understand how animals adapt to and exploit their environment. In this work we analysed the 87Sr/86Sr, δ18O and δ13C isotope composition of 65 moose bone and antler samples from Sweden from wild-shot individuals dated between 1800 and 1994 to study moose mobility and feeding behaviour for (paleo)ecological applications. Sr data were compared with isoscapes of the Scandinavian region, built ad-hoc during this study, to understand how moose utilise the landscape in Northern Europe. The 87Sr/86Sr isoscape was developed using a machine-learning approach with external geo-environmental predictors and literature data. Similarly, a δ18O isoscape, obtained from average annual precipitation δ18O values, was employed to highlight differences in the isotope composition of the local environment vs. bone/antler. Overall, 82% of the moose samples were compatible with the likely local isotope composition (n = 53), suggesting that they were shot not far from their year-round dwelling area. 'Local' samples were used to calibrate the two isoscapes, to improve the prediction of provenance for the presumably 'non-local' individuals. For the latter (n = 12, of which two are antlers and ten are bones), the probability of geographic origin was estimated using a Bayesian approach by combining the two isoscapes. Interestingly, two of these samples (one antler and one bone) seem to come from areas more than 250 km away from the place where the animals were hunted, indicating a possible remarkable intra-annual mobility. Finally, the δ13C data were compared with the forest cover of Sweden and ultimately used to understand the dietary preference of moose. We interpreted a difference in δ13C values of antlers (13C-enriched) and bones (13C-depleted) as a joint effect of seasonal variations in moose diet and, possibly, physiological stresses during winter-time, i.e., increased consumption of endogenous 13C-depleted lipids.

Molecular and carbon isotopic evidence for the possible origin of 13α(n-alkyl)-tricyclic terpanes in Mesoproterozoic bitumens from the Yanliao Rift, North China

The Mesoproterozoic Jixian System in the Yanliao Rift of the North China Craton contains organic-rich shales within the Hongshuizhuang, Tieling, and Xiamaling formations. The biomarker composition of these formations is characterized by the dominance of 13α(n-alkyl)-tricyclic terpanes. The distinctive structure and main occurrence of 13α(n-alkyl)-tricyclic terpanes in the Mesoproterozoic Jixian System in the Yanliao Rift have intrigued researchers, raising questions about their origin and formation mechanisms. This study found that δ13C values of 13α(n-alkyl)-tricyclic terpanes ranged from −32.7‰ to −26.2‰, similar to those of kerogen and n-alkanes, but significantly more depleted in 13C compared to 13β(H),14α(H)-tricyclic terpanes, which ranged from −28.5‰ to −25.7‰. The difference in δ13C values suggested that 13α(n-alkyl)-tricyclic terpanes were not formed by the same mechanism or precursors as 13β(H),14α(H)-tricyclic terpanes. No sterane biomarkers indicative of eukaryotic source input were detected. Instead, the comparable concentrations of 13α(n-alkyl)-tricyclic terpanes and n-alkanes in the samples implied a possible origin from primary organic matter producers, notably cyanobacteria. Moreover, the biomarkers obtained from distinct formations within the Mesoproterozoic Jixian System exhibited an impressive degree of simplicity, similarity and mutual correlation. These findings collectively suggest the prevalence of a cyanobacteria-dominated primitive ecosystem during the Mesoproterozoic Era within the confines of the Yanliao Rift.

Potential Residence and Coexistence Strategy of Tursiops truncatus in a Coastal Lagoon in the Southern Gulf of Mexico: Ecological Inferences Using Stable Isotopes

The Terminos Lagoon Flora and Fauna Protection Area is the largest coastal lagoon in the southern Gulf of Mexico and has the largest population of bottlenose dolphins, Tursiops truncatus, reported in the region. Photo identification studies have reported permanent and seasonal residence of dolphins; however, how dolphins coexist ecologically is unknown. In the present study, from 17 dead specimens stranded on the coast of Isla del Carmen (barrier island) between 2014 and 2016, stable isotopes in the liver, heart, and muscle, which represent different time scales, were analyzed to determine residence through δ13C values and niche width and trophic coexistence through δ15N values. Differences between sexes (δ13C liver: p = 0.009; heart: p = 0.01) and age classes (δ15N muscle: p = 0.02) were found, which indicated trophic sexual segregation and changes in resource use during growth, respectively. The lack of significant differences in δ13C values indicated isotopic equilibrium between the tissues, indicating the possible permanent residence of 12 dolphins in Terminos Lagoon as a coastal ecotype (− 16‰) and a second group (five dolphins) as an oceanic ecotype (− 18‰). In conclusion, most dolphins stranded in the Terminos Lagoon are potential permanent residents of this habitat, with evidence of resource partitioning as a coexistence strategy.

Spatial patterns of resource subsidies in Great Lakes tributaries from migratory fishes

Migratory fish can influence the stream ecosystems where they spawn by depositing large amounts of energy rich eggs, carcasses, and excrement which can increase primary and secondary productivity. Past research in the Great Lakes has focused on individual tributaries; accordingly, there is a poor understanding of how wide-spread resource subsidies are amongst tributaries. To determine which Great Lake tributaries received subsidies, we compared carbon stable isotope values (δ13C) of resident stream fishes above and below 54 barriers (e.g., low-head lamprey weirs, natural waterfalls) in 31 tributaries. Subsidies, as indicated by the difference in δ13C values above and below barriers, were common. The magnitude of the subsidy varied among Great Lakes with Lake Superior having significantly larger subsidies than the other three lakes. Barrier type (complete, partial) was not related to subsidy magnitude; however, the classification of barrier type was imperfect. Correspondence analysis of regional and local environmental factors showed that large subsidies were associated with oligotrophic streams with steep channels on the Canadian Shield with little agricultural land. The association between subsidies and canopy cover and substrate size were poor. Migratory fish supply resource subsidies to rivers across the Great Lakes basin, though their magnitude is contextually dependent. The presence of barriers are likely limiting the production of migratory fishes that depend on streams for juvenile production. The importance of resource subsidies should be considered when decisions are made about the fate of existing, and the construction of new, barriers that may reduce stream productive capacity.

Petroleum Retention, Intraformational Migration and Segmented Accumulation within the Organic‐rich Shale in the Cretaceous Qingshankou Formation of the Gulong Sag, Songliao Basin, Northeast China

AbstractIn this study, organic geochemical and petrological analyses were conducted on 111 shale samples from a well to understand the retention, intraformational migration and segmented accumulation (shale oil enrichment in different intervals is unconnected) features of shale oil within the organic‐rich shale in the Qingshankou Formation of the Gulong Sag. Our study shows that retained petroleum characteristics in the investigated succession are mainly influenced by three factors: organic richness, intraformational migration and segmented accumulation. Organic matter richness primarily controls the amount of retained petroleum, especially the ‘live’ component indicated by the S2 value rather than the total organic carbon (TOC) figure alone. The negative expulsion efficiencies determined by mass‐balance calculations of hydrocarbons reveal that petroleum from adjacent organic‐rich intervals migrates into the interval of about 2386–2408 m, which is characterized by high free hydrocarbon (S1), OSI and saturated hydrocarbons content, along with a greater difference in δ13C values between polar compounds (including resins and asphaltenes) and saturated hydrocarbons. The depth‐dependent heterogeneity of carbon isotope ratios (δ13C) of mud methane gas, δ13C of extracts gross composition (SARA), δ13C of kerogen and SARA content of extracts suggest that the studied succession can be subdivided into four intervals. The shale oil sealing enrichment character in each interval is further corroborated by the distinct δ13C values of mud methane gas in different intervals. Due to the migration of petroleum into the 2386–2408 m interval, the S1, OSI and saturated hydrocarbons content of the interval show higher relative values. The maturity of organic matter in the 2471–2500 m interval is at the highest with the smaller size molecular components of the retained petroleum. Thus, favorable ‘sweet spots’ may be found in the 2386–2408 m interval and the 2471–2500 m interval, according to the experiment results in this study.

A year-round observation of δ13C of dicarboxylic acids and related compounds in fine aerosols: Implications from Central European background site

Isotopic analysis of specific compounds in aerosols can be a useful tool when studying atmospheric processes. Here, we present the results of stable carbon isotope ratio (δ13C) measurements performed on a one-year set (n = 96, Sep. 2013–Aug. 2014) of dicarboxylic acids and related compounds in PM1 at a rural Central European background site, Košetice (Czech Republic).The most 13C enriched acid was oxalic (C2, annual average = −16.6 ± 5.0‰) followed by malonic (C3, avg. = −19.9 ± 6.6‰) and succinic (C4, avg. = −21.3 ± 4.6‰) acids. Thus, δ13C values decreased with an increase in carbon numbers. Azelaic acid (C9, avg. = −27.2 ± 3.6‰) was found to be the least 13C enriched.A comparison of δ13C of dicarboxylic acids from other background sites, especially in Asia, shows similar values to those from the European site. This comparison also showed that C2 is more 13C enriched at background sites than at urban ones. In general, we did not observe significant seasonal differences in δ13C values of dicarboxylic acids at the Central European station. We observed statistically significant differences (p value < 0.05) between winter and summer δ13C values solely for C4, glyoxylic acid (ωC2), glutaric acid (C5) and suberic acid (C8).The only significant correlations between δ13C of C2 and δ13C of C3 were found in spring and summer, suggesting that the oxidation of C3 to C2 is significant in these months with a strong contribution from biogenic aerosols. The strongest season-independent annual correlation was observed in δ13C values between C2 and C4, the two dominant dicarboxylic acids. Therefore, C4 appears to be the main intermediate precursor of C2 throughout the whole year.

Evaluation of honey authenticity in Thailand by analysis of carbon stable isotope ratio using elemental analyser coupled to isotope ratio mass spectrometry and cavity ring‐down spectrometry

SummaryIn Thailand, honey is used in a variety of commercial products that are distributed in domestic markets and abroad. As honey in Thailand comes from many locations, it is challenging to control the production quality of commercial products. The main problem commonly found in the honey industry is adulteration with undeclared added sugars. Identification of adulterated honey is crucial for authentication, which can increase consumer confidence on honey purity. In this study, 49 honey samples from beekeepers (natural) and 15 commercial honey producers were collected from different cities across Thailand. They were analysed for stable carbon isotope ratios (expressed as 13C/12C or δ13C and reported in relation to the international reference standard unit Vienna Pee Dee Belemnite, ‰ VPDB) using Elemental Analysis Isotope Ratio Mass Spectrometry (EA‐IRMS) for developing a δ13C database of natural honey and for checking the adulteration of honey. The δ13C values detected in 49 natural honey samples varied from −23.1‰ to −29.1‰ VPDB with an average of −26.8 ± 1.2‰ VPDB. According to the AOAC standard (No. 998.12, 2019), natural honey should have δ13C values lower than −23.5‰ VPDB, with an acceptable difference in δ13C values between honey and the protein extracted from honey up to 1‰. Compared to the standard guideline, 60% of commercial honey samples collected for this study (9 out of 15 samples) were found to contain undeclared added sugars.

Water use efficiency responses to fluctuating soil water availability in contrasting commercial sugar beet varieties.

Many areas of sugar beet production will face hotter and drier summers as the climate changes. There has been much research on drought tolerance in sugar beet but water use efficiency (WUE) has been less of a focus. An experiment was undertaken to examine how fluctuating soil water deficits effect WUE from the leaf to the crop level and identify if sugar beet acclimates to water deficits to increase WUE in the longer term. Two commercial sugar beet varieties with contrasting upright and prostrate canopies were examined to identify if WUE differs due to contrasting canopy architecture. The sugar beet were grown under four different irrigation regimes (fully irrigated, single drought, double drought and continually water limited) in large 610 L soil boxes in an open ended polytunnel. Measurements of leaf gas exchange, chlorophyll fluorescence and relative water content (RWC) were regularly undertaken and stomatal density, sugar and biomass yields and the associated WUE, SLW and Δ13C were assessed. The results showed that water deficits generally increase intrinsic (WUEi) and dry matter (WUEDM) water use efficiency but reduce yield. Sugar beet recovered fully after severe water deficits, as assessed by leaf gas exchange and chlorophyll fluorescence parameters and, except for reducing canopy size, showed no other acclimation to drought, and therefore no changes in WUE or drought avoidance. Spot measurements of WUEi, showed no differences between the two varieties but the prostrate variety showed lower Δ13C values, and traits associated with more water conservative phenotypes of a lower stomatal density and greater leaf RWC. Leaf chlorophyll content was affected by water deficit but the relationship with WUE was unclear. The difference in Δ13C values between the two varieties suggests traits associated with greater WUEi may be linked to canopy architecture.

Stable isotope profiles of soil organic carbon in forested and grassland landscapes in the Lake Alaotra basin (Madagascar): insights in past vegetation changes

Abstract. The extent to which the central highlands of Madagascar were once covered by forests is still a matter of debate: while reconstructing past environments is inherently difficult, the debate is further hampered by the fact that the evidence documenting land cover changes and their effects on carbon and sediment dynamics in Madagascar has hitherto mainly been derived from lake coring studies. Such studies provide an integrated view over relatively large areas but do not provide information on how land-use change affects hillslopes in terms of carbon and sediment dynamics. Such information would not only be complementary to lake inventories but may also help to correctly interpret lake sediment data. Carbon stable isotope ratios (δ13C) are particularly useful tracers to study the past dynamics of soil carbon over time spans ranging from years to millennia and thus to understand the consequences of land-use change over such time spans. We analysed soil profiles down to a depth of 2 m from pristine forests and grasslands in the Lake Alaotra region in central Madagascar. Along grassland hillslopes, soil organic carbon (SOC) content was low, from 0.4 % to 1.7 % in the top layer, and decreased rapidly to ca. 0.2 % below 100 cm depth. The current vegetation predominantly consists of C4 grasses (δ13C ∼ −13 ‰), yet topsoil δ13C-OC ranges between −23.0 ‰ and −15.8 ‰, and most profiles show a decrease in δ13C-OC with depth. This contrasts with our observations in the C3-dominated forest profiles, which show a typical profile whereby δ13C values increase slightly with depth. Moreover, the SOC stock of grasslands was ∼ 55.6 % lower than along the forested hillslopes for the upper 0–30 cm layer. δ13C values in grassland and forest profiles converge to similar values (within 2.0 ± 1.8 ‰) at depths below ∼ 80 cm, suggesting that the grasslands in the Lake Alaotra region have indeed developed on soils formerly covered by a tree vegetation dominated by C3 plants. We also observed that the percent of modern carbon (pMC) of the bulk OC in the top, middle and lower middle positions of grasslands was less than 85 % near the surface. This could reflect a combination of (i) the long residence time of forest OC in the soil, (ii) the slow replacement rate of grassland-derived OC (iii) and the substantial erosion of the top positions towards the valley position of grasslands. At the valley positions under grassland, the upper 80 cm contains higher amounts of recent grass-derived OC in comparison to the hillslope positions. This is likely to be related to the higher productivity of the grassland valleys (due to higher moisture and nutrient availability), and the deposition of OC that was eroded further upslope may also have contributed. The method we applied, which is based on the large difference in δ13C values between the two major photosynthetic pathways (C3 and C4) in (sub-)tropical terrestrial environments, provides a relatively straightforward approach to quantitatively determine changing vegetation cover, and we advocate for its broader application across Madagascar to better understand the island's vegetation history.

Stable isotope differences of polar bears in the Southern Beaufort Sea and Chukchi Sea

AbstractThe life‐history, genetic, and habitat use differences between the 2 polar bear (Ursus maritimus) subpopulations in Alaska, USA, have been used to determine the geographic border separating them, but it has sparked a debate of the correct placement of the border for several years. Recently, the Southern Beaufort Sea (SBS) polar bear subpopulation has declined because of sea ice loss, while the Chukchi Sea (CS) subpopulation appears stable. To provide additional information about potential differences between the SBS and CS subpopulations, such as differences in prey sources, we used stable isotope analysis of carbon and nitrogen from bone collagen of polar bears in these 2 neighboring subpopulations. We analyzed polar bear bones from 112 individuals collected from 1954–2019. Our purpose was to determine if the SBS and CS subpopulations could be distinguished based on the stable isotope signatures of bone collagen. A difference &gt;1‰ in stable carbon isotope (δ13C) values suggests a change in carbon sources, such as nearshore to offshore, while a 3‰ change in stable nitrogen isotope (δ15N) values equates to a change of about 1 trophic level. Our study indicated a difference in δ13C values (P ≤ 0.001) but not δ15N values (P = 0.654) between the CS (−13.0 ± 0.3‰ and 22.0 ± 0.9‰, respectively) and SBS bears (−14.7 ± 1.3‰ and 22.2 ± 1.0‰, respectively). Our findings indicate that the 2 subpopulations are consuming similar high trophic level prey, while feeding in ecosystems with different δ13C baselines. We performed a logistic regression analysis using δ13C and δ15N values of the polar bears to predict their placement into these 2 subpopulations. Using Icy Cape, Alaska as the geographical boundary, the analysis correctly placed polar bears in their respective subpopulations 82% of the time. Overall accuracy of placement changed to 84% when using the current geographical boundary at Utqiaġvik, Alaska. We predicted samples collected from the Wainwright, Alaska region as 58% CS and 42% SBS polar bears. This suggests that the area between Wainwright and Icy Cape is a polar bear mixing zone that includes bears from both subpopulations. Bone collagen has a long‐term, potentially life‐long, stable isotope turnover rate, and our results could be used to determine the association of harvested polar bears to Alaska subpopulations, thus aiding in transboundary harvest quota management.

Mercury biomagnification in an Antarctic food web of the Antarctic Peninsula

Under the climate change context, warming Southern Ocean waters may allow mercury (Hg) to become more bioavailable to the Antarctic marine food web (i.e., ice-stored Hg release and higher methylation rates by microorganisms), whose biomagnification processes are poorly documented. Biomagnification of Hg in the food web of the Antarctic Peninsula, one of the world's fastest-warming regions, was examined using carbon (δ13C) and nitrogen (δ15N) stable isotope ratios for estimating feeding habitat and trophic levels, respectively. The stable isotope signatures and total Hg (T-Hg) concentrations were measured in Antarctic krill Euphausia superba and several Antarctic predator species, including seabirds (gentoo penguins Pygoscelis papua, chinstrap penguins Pygoscelis antarcticus, brown skuas Stercorarius antarcticus, kelp gulls Larus dominicanus, southern giant petrels Macronectes giganteus) and marine mammals (southern elephant seals Mirounga leonina). Significant differences in δ13C values among species were noted with a great overlap between seabird species and M. leonina. As expected, significant differences in δ15N values among species were found due to interspecific variations in diet-related to their trophic position within the marine food web. The lowest Hg concentrations were registered in E. superba (0.007 ± 0.008 μg g−1) and the highest values in M. giganteus (12.090 ± 14.177 μg g−1). Additionally, a significant positive relationship was found between Hg concentrations and trophic levels (reflected by δ15N values), biomagnifying nearly 2 times its concentrations at each level. Our results support that trophic interaction is the major pathway for Hg biomagnification in Southern Ocean ecosystems and warn about an increase in the effects of Hg on long–lived (and high trophic level) Antarctic predators under climate change in the future.

Archaeal tetraether lipids and their biphytane carbon isotope composition in sediments along an estuarine biogeochemical gradient

The stable carbon isotopic composition of Thaumarchaeota-produced crenarchaeol (δ13Ccren) is thought to reflect the δ13C of dissolved inorganic carbon (DIC) and is valuable in carbon cycle research. Recently, an isotope flux-balance model was proposed that δ13Ccren is dependent on passive CO2 uptake and a slow rate of intracellular conversion to HCO3−, which has been validated in open ocean environments. However, further studies are required in environments with larger ranges of aqueous CO2 and diverse archaeal communities to examine whether the δ13Ccren value could still be predicted by the model. Here, we report δ13C values of biphytanes chemically released from the archaeal tetraether lipids (i.e., GDGTs) in sediments along a transect exhibiting shifts in the archaeal community and showing gradients of salinity, pH, and CO2 in the Pearl River Estuary, South China. Methanogenic and methanotrophic Euryarchaeota are present in the upper estuary, although not significant, as revealed by the elevated relative abundances of GDGT-0 and higher values of methane index in suspended particulate matter (SPM) and surface sediments, as well as the more 13C-depleted C40:0, C40:1, and C40:2 biphytanes than the C40:3 biphytane released from sedimentary GDGTs. Toward the lower estuary and coastal sea, GDGT distributions in SPM and sediments tend to indicate archaeal communities under normal marine conditions. However, there are still differences in δ13C values between C40:0 and C40:3 biphytanes by up to 2.4‰ in sediments, although C40:1, C40:2, and C40:3 were similar in δ13C, likely due to the existence of marine group II Euryarchaeota. Along such a physicochemical and biogeochemical gradient, the values of sedimentary δ13C40:3 increase monotonically with the increase of mean annual pH and the decrease of mean annual CO2. This phenomenon resembles the reported δ13CGDGT profiles, along with the variations in pH and CO2, in water column in the open ocean, and hence consistent in direction with the isotope flux-balance model for Thaumarchaeota lipids. However, our estimated apparent biosynthetic isotope effect (εAr) of crenarchaeol, ranging from 15.2‰ to 19.7‰, is mostly lower than those estimated in the open ocean. With the constraints of thaumarchaeal growth rates in such a dynamic estuarine environment as well as those in the open ocean, we suggest slight reductions in catalytic efficiency and intracellular kinetic isotope effect of intracellular CO2 hydration relative to previously employed values. Such modifications would slightly change the sensitivities of εAr to archaeal growth rate and ambient CO2 concentration but might be more suitable for variable hydrology and community environments.

Mycobiont diversity and first evidence of mixotrophy associated with Psathyrellaceae fungi in the chlorophyllous orchid Cremastra variabilis.

Mixotrophy (MX, also called partial mycoheterotrophy) in plants is characterized by isotopic abundances that differ from those of autotrophs. Previous studies have evaluated mycoheterotrophy in MX plants associated with fungi of similar ecological characteristics, but little is known about the differences in the relative abundances of 13C and 15N in an orchid species that associates with several different mycobionts species. Since the chlorophyllous orchid Cremastra variabilis Nakai associates with various fungi with different ecologies, we hypothesized that it may change its relative abundances of 13C and 15N depending on the associated mycobionts. We investigated mycobiont diversity in the chlorophyllous orchid C. variabilis together with the relative abundance of 13C and 15N and morphological underground differentiation (presence or absence of a mycorhizome with fungal colonization). Rhizoctonias (Tulasnellaceae, Ceratobasidiaceae, Sebacinales) were detected as the main mycobionts. High differences in δ13C values (-34.7 to -27.4 ‰) among individuals were found, in which the individuals associated with specific Psathyrellaceae showed significantly high relative abundance of 13C. In addition, Psathyrellaceae fungi were always detected on individuals with mycorhizomes. In the present study, MX orchid association with non-rhizoctonia saprobic fungi was confirmed, and the influence of mycobionts on morphological development and on relative abundance of 13C and 15N was discovered. Cremastra variabilis may increase opportunities to gain nutrients from diverse partners, in a bet-hedging plasticity that allows colonization of various environmental conditions.

Mass spectrometry and isotopes analysis of the saturated fraction biomarkers

GC-IRMS analysis extends and confirms (or not) the interpretation based on the results of GC-MS analyses. For example, it is very useful in determining the sedimentation environment of organic matter. GC-MS analysis of biomarkers and the results are reliable, but only GC-IRMS studies can confirm it. In this study, the origin of BNH (28,30-bisnorhopane from chemoautotrophic bacteria) and origin of higher carotenoids and their derivatives from Chlorobiaceae or Chromotiaceae bacteria were confirmed through isotopic analyzes. Biomarkers were analyzed using the GC-IRMS and EA-IRMS apparatus. The obtained chromatograms from the IRMS analyses were compared with the archival GC-MS analyses for the same samples in order to identify individual chemical compounds. In addition to the existing methodology of sample preparation for analyses, a non-standard method was also used, consisting in the separation of n-alkanes from branched hydrocarbons. The repeatability of the method was determined on the GC-IRMS and the values of δ13C for selected biomarkers from the saturated fraction were determined. It was found that samples with low biomarker content are not suitable for analysis. On the other hand, too high concentration of the analyte causes an increase of the chromatogram baseline and worse separation of the peaks, which is also a problem. For the crude oils the δ13C values were initially determined for the biomarkers of the saturated fraction from the hopanes group: bisnorhopane (BNH), oleanane, C29 norhopane, C30 hopane, moretane and the C31-C35 homohopane series. Relatively small differences in δ13C values were found between BNH/hopanes and BNH/crude oils, which suggests the same source of origin for all biomarkers (including BNH). Determining biomarkers in the aromatic fraction using the GC-IRMS method was not successful. In the future, a special methodology for preparing samples for carbon isotopic analyses of aromatic fraction will be required.

Lipids and their δ13C values reveal carbon assimilation and cycling in the methane-seep tubeworm Paraescarpia echinospica from the South China Sea

Gutless tubeworms obtain their nutrition from their bacterial endosymbiotic partners, and the metazoan hosts provide the bacteria with sulfide, carbon dioxide, and molecular oxygen taken up through the plume. At seeps, tubeworms take up sulfide using their posterior end, allowing them to fix carbon dioxide and synthesize organic molecules. Compared to bulk tissue isotope compositions, lipid biomarkers and their compound-specific δ13C values of soft tissues allow to assign nutritional modes of endosymbionts and hosts, as well as associations between endosymbionts and the animal host. In this study, the tubeworm Paraescarpia echinospica retrieved from the Haima seeps in the South China Sea (SCS) at approximately 1400 m water depth was analyzed using 16S rRNA, bulk tissue isotopes, lipid biomarkers, and compound-specific δ13C values. The Lamellibrachia sp. endosymbiont, a phylotype of the Gammaproteobacteria, is a bacterium gaining energy from oxidizing sulfur compounds (thiotrophic endosymbiont). Further, six phylotypes of Gammaproteobacteria and seven phylotypes of Alphaproteobacteria were identified in the trophosome. High contents of n-C16:1ω7, n-C18:1ω7 fatty acids, and n-C16:0 and n-C18:1ω7 fatty alcohols were found in the trophosome, and somewhat lower contents in the plume and vestimentum, consistent with the prevalence of thiotrophic endosymbionts in the trophosome. The fatty acids n-C16:1ω7 and n-C18:1ω7 produced by the endosymbionts yielded δ13C values ranging from −34‰ to −30‰, reflecting the use of marine dissolved inorganic carbon (DIC) rather than DIC derived from anaerobic oxidation of methane. The overall small differences in δ13C values (Δamong tissues ≤3‰) for n-C16:1ω7 and n-C18:1ω7 fatty acids in both symbiont-containing and symbiont-free tissues appear to represent the δ13C signatures of the endosymbionts. Generally, n-C16:1ω7 and n-C18:1ω7 fatty acids should be more 13C-depleted than the respective n-C16:0, n-C18:0 fatty acids resulting from desaturation. However, in this study, the saturated n-C16:0, n-C18:0 fatty acids in soft tissues were 13C depleted by 4‰–10‰ compared to the respective n-C16:1ω7 and n-C18:1ω7 fatty acids, suggesting complex trophic interactions between P. echinospica, its endosymbionts, other bacteria living in P. echinospica, and the environment. Sterols, yielding the lowest δ13C values from −49‰ to −46‰, are interpreted to be derived from bioconversion of dietary sterols by the animal host. The combination of microbial diversity, bulk tissue isotopes, and lipid biomarkers used here is a valuable approach to evaluate the nutritional associations between symbionts and hosts in chemosymbiosis as well as the diet of the animal host.

Stable isotopes for reliable identification of wild and mass-reared Queensland fruit flies in sterile insect technique programs

Queensland fruit fly is one of the most economically important horticultural pests in Australia. Sterile insect technique (SIT) is now being reconsidered and upscaled to combat this pest so reliable discrimination of released sterile Q-flies from wild flies in monitoring traps is important for effective SIT operations. Stable isotopes provide a permanent chemical marker to discriminate sterile and wild flies when dye marking is unclear. In this study, we compared the isotopic ratios of carbon and nitrogen between Q-flies reared on different larval diets and wild flies collected from diverse locations in Australia and New Caledonia. Finally, we conducted a release–recapture study to corroborate differences in stable isotope C and N ratios in laboratory-reared and wild Q-flies. The δ15N values obtained from wild and laboratory Q-flies showed high variability that is likely related to the food source of the larval and/or adult stage and do not offer an effective means to discriminate between sterile and wild Q-flies. The δ13C values of examined wild Q-flies ranged from − 27.46 to − 24.37‰ VPDB, whereas those from laboratory-reared, released and recaptured Q-flies ranged from − 25.73 to − 19.26‰ VPDB. Differences in δ13C values resulted in 100% correct classification of wild flies and 96.88% correct classification of released flies. Measurements of intrinsic δ13C values offer a precise tool to discriminate between sterile and wild Q-flies in SIT programs, regardless of the composition of the larval or adult pre-release diets.

Assessment of the influence of sample preparation on the results of pyrolysis analyses

The aim of the work, the results of which are presented in the article, was to evaluate the influence of the method of sample preparation on the results of pyrolytic analyses: RockEval, Py-GC-FID (gas chromatography with FID detector) and Py-GC-IRMS (isotope mass spectrometry with gas chromatography). Mortars and ball mills are most often used to grind the samples. Three rock samples with a higher organic carbon content (shales) and five with a lower organic carbon content (including two samples of dolomites and three of anhydrites) were prepared. The rocks were homogenized and divided into three parts. Each part was ground: by hand in a mortar, in a ball mill for 5 minutes, and in a ball mill for 15 minutes. A total of 24 RockEval analyses, 48 Py-GC-FID analyses (two pyrolysis temperatures corresponding to RockEval pyrolysis conditions) and 24 Py-GC-IRMS analyses were performed. The grinding of the samples for the RockEval analyses is insignificant. The differences in the results seem to indicate the influence of the analysis error and the nature of the sample. For the Py-GC-FID methodology, the influence of milling on the results of desorption carried out at the temperature of 300ºC for most of the tested samples can be considered insignificant and negligible. At the temperature of 500ºC, various types of samples show some correlations, but they are insufficient to reject one of the methods of sample preparation. Therefore, it is important to analyze the results comprehensively, taking into account both the group composition and the distribution of pyrolysis products. In the case of the isotopic composition analyses (Py-GC-IRMS), also no differences in δ13C values related to different types of grinding samples can be found. Summarizing, all the results show a negligible influence of the method of grinding the samples on the results of pyrolysis analyses. Nevertheless, it is important that only one method of sample preparation is used for a separate sample series.

Isotopic niche partitioning between two small cetacean species

Commerson’s dolphin Cephalorhynchus commersonii and Peale’s dolphin Lagenorhynchus australis live in sympatry along the southwestern South Atlantic Ocean, suggesting the existence of some degree of habitat partitioning to reduce their competition for resources. Both species are usually associated with coastal environments, but information on their trophic ecology is scarce. Here we explored the existence of trophic resource partitioning between these 2 sympatric species using δ13C and δ15N. From 2007 to 2013, we analyzed a total of 14 Commerson’s dolphin and 34 Peale’s dolphin skin samples from a marine protected area in Argentina. Significant differences in δ13C values indicated intra- and interspecific spatial partitioning. Three different Peale’s dolphin feeding groups (FGs) were identified: FG1 exclusively exploited the pelagic habitat, FG2 occupied more pelagic/inshore habitats (similar to Commerson’s dolphins), and FG3 occupied more benthic/inshore habitats. Isotopic niche breadth varied between species and feeding groups, exhibiting less variation in prey selection among Commerson’s dolphins. According to Bayesian standard ellipse area analysis, isotopic niche overlap was found between FG2 and FG3 from 2007 to 2012, and between FG2 and Commerson’s dolphins. Mixing models suggest that FG3 has a diet mainly based in benthic fish, while FG2 and Commerson’s dolphins feed predominantly on pelagic fish. Overall, this study indicates segregation in the use of trophic resources between 2 sympatric dolphin species, showing different foraging strategies that promote coexistence and reduce intra- and interspecific competition. Furthermore, the presence of 3 Peale’s dolphin feeding groups segregated at a small geographic scale suggests a previously unknown ecological complexity.

Two-stage mineral dissolution and precipitation related to organic matter degradation: Insights from in situ C–O isotopes of zoned carbonate cements

Mineral dissolution and precipitation, particularly of carbonate minerals, are ubiquitous and significant diagenetic processes in sedimentary rocks. However, the impacts of organic matter degradation on these processes in single systems remain unclear because the timing and fluid sources during progressive burial are poorly constrained. To address this issue, in situ C–O isotope analyses were conducted for zoned dolomite–ankerite/calcite cements from the Permian Lucaogou Formation in the Junggar Basin, China, using secondary ion mass spectrometry (SIMS) combined with mineralogical and elemental analyses. Carbonate minerals can be divided into early-stage and late-stage precipitations based on distinct variations in δ18O (−18.3 to −0.7‰ V-PDB), δ13C (−3.6 to +20.8‰ V-PDB), and Fe content (Fe# ranging from 0.002 to 0.373). The chemo-isotopically zoned carbonate cements record the thermal and chemical conditions of pore fluids during the burial process. The difference in δ18O values between the early and late precipitation (i.e., Δ18O [early–late]) of 11.9‰, and the difference in δ13C values (i.e., Δ13C [early–late]) of 11.7‰, reveal significant changes in precipitation temperatures and the availability of carbon sources, respectively. Early-stage precipitation occurred in association with microbial methanogenesis at lower temperatures (~25 °C) during shallow burial (~300 m), whereas late-stage precipitation was related to thermally-induced decarboxylation at higher temperatures (~90 °C or ~110 °C) during deep burial (~2500 m or ~3200 m). These two stages of dissolved pore-filling precipitation correspond to two major periods of dissolution (i.e., eogenetic and mesogenetic dissolution). Diagenetic fluids, mainly CO2 and/or organic acids, were derived from the biological and thermal degradation of organic matter, respectively. These results constrain the isotopic responses of carbonates to organic matter degradation with increasing burial depth and provide significant insights into carbonate diagenesis in organic-rich sedimentary sequences.

Communities of mycorrhizal fungi in different trophic types of Asiatic Pyrola japonica sensu lato (Ericaceae).

Mixotrophic plants obtain carbon by their own photosynthetic activity and from their root-associated mycorrhizal fungi. Mixotrophy is deemed a pre-adaptation for evolution of mycoheterotrophic nutrition, where plants fully depend on fungi and lose their photosynthetic activity. The aim of this study was to clarify mycorrhizal dependency and heterotrophy level in various phenotypes of mixotrophic Pyrola japonica (Ericaceae), encompassing green individuals, rare achlorophyllous variants (albinos) and a form with minute leaves, P. japonica f. subaphylla. These three phenotypes were collected in two Japanese forests. Phylogenetic analysis of both plants and mycorrhizal fungi was conducted based on DNA barcoding. Enrichment in 13C among organs (leaves, stems and roots) of the phenotypes with reference plants and fungal fruitbodies were compared by measuring stable carbon isotopic ratio. All plants were placed in the same clade, with f. subaphylla as a separate subclade. Leaf 13C abundances of albinos were congruent with a fully mycoheterotrophic nutrition, suggesting that green P. japonica leaves are 36.8% heterotrophic, while rhizomes are 74.0% heterotrophic. There were no significant differences in δ13C values among organs in both albino P. japonica and P. japonica f. subaphylla, suggesting full and high mycoheterotrophic nutrition, respectively. Among 55 molecular operational taxonomic units (OTUs) detected as symbionts, the genus Russula was the most abundant in each phenotype and its dominance was significantly higher in albino P. japonica and P. japonica f. subaphylla. Russula spp. detected in P. japonica f. subaphylla showed higher dissimilarity with other phenotypes. These results suggest that P. japonica sensu lato is prone to evolve mycoheterotrophic variants, in a process that changes its mycorrhizal preferences, especially towards the genus Russula for which this species has a marked preference.