Investigation of the microbial metabolism of carbon dioxide and hydrogen in the kangaroo foregut by stable isotope probing

Kangaroos ferment forage material in an enlarged forestomach analogous to the rumen, but in contrast to ruminants, they produce little or no methane. The objective of this study was to identify the dominant organisms and pathways involved in hydrogenotrophy in the kangaroo forestomach, with the broader aim of understanding how these processes are able to predominate over methanogenesis. Stable isotope analysis of fermentation end products and RNA stable isotope probing (RNA-SIP) were used to investigate the organisms and biochemical pathways involved in the metabolism of hydrogen and carbon dioxide in the kangaroo forestomach. Our results clearly demonstrate that the activity of bacterial reductive acetogens is a key factor in the reduced methane output of kangaroos. In in vitro fermentations, the microbial community of the kangaroo foregut produced very little methane, but produced a significantly greater proportion of acetate derived from carbon dioxide than the microbial community of the bovine rumen. A bacterial operational taxonomic unit closely related to the known reductive acetogen Blautia coccoides was found to be associated with carbon dioxide and hydrogen metabolism in the kangaroo foregut. Other bacterial taxa including members of the genera Prevotella, Oscillibacter and Streptococcus that have not previously been reported as containing hydrogenotrophic organisms were also significantly associated with metabolism of hydrogen and carbon dioxide in the kangaroo forestomach.The ISME Journal advance online publication, 13 March 2014; doi:10.1038/ismej.2014.25.

Stable isotope changes in freshwater shrimps (Exopalaemon modestus and Macrobrachium nipponensis): trophic pattern implications

Trophic patterns of omnivorous freshwater shrimps, Exopalaemon modestus and Macrobrachium nipponensis, were investigated in two shallow eutrophic lakes by using stable isotope analysis. delta N-15 and delta C-13 of M. nipponensis and E. modestus increased with increasing body weight, which might be attributed to larger individuals ingesting organisms that feed higher up the food chain and/or increased assimilation of benthic food items with enriched isotopic signatures. Of the freshwater shrimps occurring in the studied lakes, those from Lake Taihu had significantly elevated delta N-15 and delta C-13 values (4.3% and 1.8%, respectively) compared with those from the less eutrophic Lake Chaohu, indicating that the isotopic signature might partially reflect the trophic states of their habitats. Mixing model results suggested that the benthic food web provides the primary carbon source for both shrimp species, and that E. modestus assimilated relatively more pelagic food sources than M. nipponensis in these lakes.

Preservation methods alter stable isotope values in gelatinous zooplankton: implications for interpreting trophic ecology

Jellyfish are increasingly topical within studies of marine food webs. Stable isotope analysis represents a valuable technique to unravel the complex trophic role of these long-overlooked species. In other taxa, sample preservation has been shown to alter the isotopic values of species under consideration, potentially leading to misinterpretation of trophic ecology. To identify potential preservation effects in jellyfish, we collected Aurelia aurita from Strangford Lough (54(o)22'44.73aEuro(3)N, 5(o)32'53.44aEuro(3)W) during May 2009 and processed them using three different methods prior to isotopic analysis (unpreserved, frozen and preserved in ethanol). A distinct preservation effect was found on delta N-15 values: furthermore, preservation also influenced the positive allometric relationship between individual size and delta N-15 values. Conversely, delta C-13 values remained consistent between the three preservation methods, conflicting with previous findings for other invertebrate, fish and mammalian species. These findings have implications for incorporation of jellyfish into marine food webs and remote sampling regimes where preservation of samples is unavoidable.

Stable isotope composition of smectite in suevites at the Ries crater, Germany: Implications for hydrous alteration of impactites

The 24-km diameter Ries crater, Germany, exhibits well-preserved crater filling and surficial melt-rich breccia deposits that are believed to have been altered by post-impact hydrothermal fluids. The alteration mineralogy of the crater filling breccias is characterized by clay (smectite, chlorite) and a zeolite assemblage, and secondary clay phases (smectite, minor halloysite) in surficial melt-bearing breccia deposits. Using stable isotope analysis of secondary smectitic clay fractions, evidence of significant hydrous alteration of impactites at large water/rock ratios was found. The estimated fluid temperatures, using data derived by delta(18)O and delta D fractionation, suggest smectite precipitation in surficial breccias in equilibrium with meteoric fluids at temperatures 16 +/- 5 degrees C in agreement with the long-term variation of modern precipitation in the area. The stable isotope composition of smectite in crater-fill breccia, however, suggests a trend of monotonously increasing temperatures from 43 to 112 degrees C. with increasing depth through the breccia sequence. This demonstrates a different origin of alteration and temperature distribution for the surficial and crater filling melt-bearing impact breccias in the Ries crater. Our results suggest that the inverted structure of hydrothermal systems observed in some terrestrial impact craters, including the Ries crater, could indicate the initial configuration of a thermal anomaly in the crater filling sequence, but which is replaced with a normal hydrothermal convection in crater proper, during the course of post-impact cooling. (C) 2010 Elsevier B.V. All rights reserved.

Salinity of the Eocene Arctic Ocean from oxygen isotope analysis of fish bone carbonate

Stable isotope analysis was performed on the structural carbonate of fish bone apatite from early and early middle Eocene samples (~55 to ~45 Ma) recently recovered from the Lomonosov Ridge by Integrated Ocean Drilling Program Expedition 302 (the Arctic Coring Expedition). The d18O values of the Eocene samples ranged from -6.84 per mil to -2.96 per mil Vienna Peedee belemnite, with a mean value of -4.89 per mil, compared to 2.77 per mil for a Miocene sample in the overlying section. An average salinity of 21 to 25 per mil was calculated for the Eocene Arctic, compared to 35 per mil for the Miocene, with lower salinities during the Paleocene Eocene thermal maximum, the Azolla event at ~48.7 Ma, and a third previously unidentified event at ~47.6 Ma. At the Azolla event, where the organic carbon content of the sediment reaches a maximum, a positive d13C excursion was observed, indicating unusually high productivity in the surface waters.

Stable isotope analyses of Cretaceous sediment samples

We report the sulfur and oxygen isotope composition of sulfate (d34SSO4 and d18OSO4, respectively) in coexisting barite and carbonate-associated sulfate (CAS), which we use to explore temporal variability in the marine sulfur cycle through the middle Cretaceous. The d34SSO4 of marine barite tracks previously reported sulfur isotope data from the tropical Pacific. The d18OSO4 of marine barite exhibits more rapid and larger isotopic excursions than the d34SSO4 of marine barite; these excursions temporally coincide with Ocean Anoxic Events (OAEs). Neither the d34SSO4 nor the d18OSO4 measured in marine barite resembles the d34SSO4 or the d18OSO4 measured in coexisting CAS. Culling our data set for elemental parameters suggestive of carbonate recrystallization (low [Sr] and high Mn/Sr) improves our record of d18OSO4 in CAS in the Cretaceous. This suggests that the CAS proxy can be impacted by carbonate recrystallization in some marine sediments. A box model is used to explore the response of the d34SSO4 and d18OSO4 to different perturbations in the marine biogeochemical sulfur cycle. We conclude that the d34SSO4 in the middle Cretaceous is likely responding to a change in the isotopic composition of pyrite being buried, coupled possibly with a change in riverine input. On the other hand, the d18OSO4 is likely responding to rapid changes in the reoxidation pathway of sulfide, which we suggest may be due to anoxic versus euxinic conditions during different OAEs.

Slow Isotope Turnover Rates and Low Discrimination Values in the American Alligator: Implications for Interpretation of Ectotherm Stable Isotope Data

Stable isotope analysis has become a standard ecological tool for elucidating feeding relationships of organisms and determining food web structure and connectivity. There remain important questions concerning rates at which stable isotope values are incorporated into tissues (turnover rates) and the change in isotope value between a tissue and a food source (discrimination values). These gaps in our understanding necessitate experimental studies to adequately interpret field data. Tissue turnover rates and discrimination values vary among species and have been investigated in a broad array of taxa. However, little attention has been paid to ectothermic top predators in this regard. We quantified the turnover rates and discrimination values for three tissues (scutes, red blood cells, and plasma) in American alligators (Alligator mississippiensis). Plasma turned over faster than scutes or red blood cells, but turnover rates of all three tissues were very slow in comparison to those in endothermic species. Alligator δ15N discrimination values were surprisingly low in comparison to those of other top predators and varied between experimental and control alligators. The variability of δ15N discrimination values highlights the difficulties in using δ15N to assign absolute and possibly even relative trophic levels in field studies. Our results suggest that interpreting stable isotope data based on parameter estimates from other species can be problematic and that large ectothermic tetrapod tissues may be characterized by unique stable isotope dynamics relative to species occupying lower trophic levels and endothermic tetrapods.

Stable Isotope Biogeochemistry of South Florida Coastal Marine Ecosystems

Southeast Florida’s continual urban expansion will potentially increase anthropogenic pollution in adjacent coastal marine systems. Furthermore, increased nutrient loads could have detrimental effects on the already threatened Florida Reef Tract. The present study uses a stable isotopic approach to determine the sources and the impact of nutrients on the Florida Reef Tract. δ13C and δ15N analysis of macroalgae, sponges, and sediment were analyzed in order to determine nutrient inputs in this region. While δ13C data did not display any significant trends spatially, δ15N values of the majority of biota exhibited a strong East to West gradient with more enriched values close to shore. Relative enrichment in δ15N values were measured for sediments sampled along the Florida Reef Tract in comparison to a pristine Marquesas Keys sediment core. The δ15N data also implies that shoreline anthropogenic nutrients have more nutrient loading implications on reefs than major point sources.

Relative contribution of natural productivity and compound feed to tissue growth in blue shrimp (Litopenaeus stylirostris) reared in biofloc: Assessment by C and N stable isotope ratios and effect on key digestive enzymes

The aim of this study was to assess the relative contribution of natural productivity and compound food to the growth of the juvenile blue shrimp Litopenaeus stylirostris reared in a biofloc system. Two experiments were carried out based on the same protocol with three treatments: clear water with experimental diet (CW), biofloc with experimental diet (BF) and biofloc unfed (BU). Shrimp survival was significantly higher in biofloc rearing than in CW rearing. The contribution of the biofloc to shrimp diet was estimated through measurement of carbon and nitrogen stable isotope ratios in shrimp and food sources. Different isotopic compositions between feeds were obtained by feeding natural productivity with a mixture rich in fish meal and the shrimps with a pellet containing a high level of soy protein concentrate. Using a two source one-isotope mixing model, we found that the natural productivity of the biofloc system contributed to shrimp growth at a level of 39.8% and 36.9%, for C and N, respectively. The natural food consumed by the shrimps reared in the biofloc system resulted in higher gene expression (mRNA transcript abundance) and activities of two digestive enzymes in their digestive gland: α-amylase and trypsin. The growth of shrimp biomass reared in biofloc was, on average, 4.4 times that of those grown in clear water. Our results confirmed the best survival and promoted growth of shrimps using biofloc technology and highlighted the key role of the biofloc in the nutrition of rearing shrimps. Statement of relevance In this study, we have applied an original protocol to determine the respective contribution of natural productivity and artificial feeds on the alimentation of the juvenile blue shrimp L. stylirostris reared in biofloc system by using C and N natural stable isotope analysis. Moreover, we have compared, in shrimp digestive gland, the α-amylase and trypsin enzyme activities at biochemical and molecular levels for two different shrimp rearing systems, biofloc and clear water. In our knowledge, the use of molecular tool to study the influence of biofloc consumption on digest process of shrimp was never carried out. We think that our research is new and important to increase knowledge on biofloc topic.

Stable isotope and multi-analytical investigation of Monte da Cegonha: A Late Antiquity population in southern Portugal

This study presents for the first time the diet of a Late Antiquity population in southern Portugal (Civitas of Pax Julia), from the Roman villa of Monte da Cegonha (predominantly 7th century CE). Stable isotope analysis (δ13C, δ15N, δ18O, 87Sr/86Sr) of human and faunal bone collagen and apatite was conducted in order to understand the influence of Roman subsistence strategies on the way of life of rural inhabitants of the area of Pax Julia and to explore their diet (types of ingested plants, amount of animal resources, terrestrial versus marine resources). X-ray diffraction (XRD) and Fourier transform infra-red spectroscopy (FTIR) analyses were used to determine the degree of bone diagenesis and assess the reliability of the bone stable isotopic composition for palaeodietary reconstruction. Anthropological analysis revealed a cariogenic diet, rich in starchy food and carbohydrates, in at least in two individuals based on the frequency of dental caries. Collagen and apatite carbon isotopic analysis suggested that C3 plants were the basis of the population's diet, complemented with some terrestrial meat and its by-products as reflected by the observed bone collagen nitrogen isotopic composition. Moreover, whilst the fairly low apatite-collagen spacing recorded in some skeletons (at around 4‰) may have been due to freshwater organisms intake, the relatively low nitrogen values observed indicate that this consumption did not occur very often, unless in the form of fresh fish of low trophic level or fish sauces. There were no significant differences in isotopic values depending on gender or burial type. Strontium and oxygen isotopic composition of bone apatite revealed a sedentary community, with the exception of a male individual who probably did not spend his childhood in Monte da Cegonha.

Improved Approach for Analyzing Bromophenols in Seafood Using Stable Isotope Dilution Analysis in Combination with SPME

An analytical method for the measurement of five naturally occurring bromophenols of sensory relevance in seafood (barramundi and prawns) is presented. The method combines simultaneous distillation−extraction followed by alkaline back extraction of a hexane extract and subsequent acetylation of the bromophenols. Analysis of the bromophenol acetates was accomplished by headspace solid phase microextraction and gas chromatography−mass spectrometry using selected ion monitoring. The addition of 13C6 bromophenol stable isotope internal standards for each of the five congeners studied permitted the accurate quantitation of 2-bromophenol, 4-bromophenol, 2,6-dibromophenol, 2,4-dibromophenol, and 2,4,6-tribromophenol down to a limit of quantification of 0.05 ng/g of fish flesh. The method indicated acceptable precision and repeatability and excellent linearity over the typical concentration range of these compounds in seafood (0.5−50 ng/g). The analytical method was applied to determine the concentration of bromophenols in a range of farmed and wild barramundi and prawns and was also used to monitor bromophenol uptake in a pilot feeding trial.

Growth and food availability of silver and bighead carps: evidence from stable isotope and gut content analysis

A 2-year investigation of growth and food availability of silver carp and bighead was carried out using stable isotope and gut content analysis in a large pen in Meiliang Bay of Lake Taihu, China. Both silver carp and bighead exhibited significantly higher delta 13C in 2005 than in 2004, which can probably be attributed to two factors: (i) the difference between isotopic compositions at the base of the pelagic food web and (ii) the difference between the compositions of prey items and stable isotopes. The significantly positive correlations between body length, body weight and stable isotope ratios indicated that isotopic changes in silver carp and bighead resulted from the accumulation of biomass concomitant with rapid growth. Because of the drastic decrease in zooplankton in the diet in 2005, silver carp and bighead grew faster in 2004 than in 2005. Bighead carp showed a lower trophic level than silver carp in 2005 as indicated by stable nitrogen isotope ratios, which was possibly explained by the interspecific difference between the prey species and the food quality of silver carp and bighead.

Contributions of Pelagic and Benthic Dietary Sources to Freshwater Mussels: Evidence from Stable Carbon Isotope Analysis

In Gonghu Bay of Lake Taihu, tissue of five mussel species showed delta C-13 values similar to or slightly below that of pelagic suspended particulate organic matter (SPOM). This indicated that mussels in this area either fed non-selectively and so reflected available carbon in the pelagic habitat or selected for phytoplankton. The situation was the same for Anodonta woodiana woodiana and Cristaria plicata in Meiliang Bay; however, for the remaining three species, Hyriopsis cumingii, Arconaia lanceolata, and Lamprotula rochechouarti, tissue had intermediate delta C-13 values, falling between those for pelagic SPOM and benthic sediment organic matter (SOM), suggesting a possible preferential selection of phytoplankton from the pelagic SPOM but more likely reflecting local differences in pelagic SPOM and benthic SOM composition and available organic carbon sources. The mixing model showed that pelagic SPOM accounted for over 98% of carbon incorporated by all mussels in Gonghu Bay and two mussels in Meiliang Bay, suggesting the dietary importance of pelagic food sources for mussels. Less than 50% of the assimilation in H. cumingii, A. lanceolata, and L. rochechouarti came from the pelagic carbon sources in Meiliang Bay, which suggested that these species consumed a mix of benthic and pelagic derived carbon sources.