Stable isotope analysis of dynamic lipidomics

Metabolic pathway flux is a fundamental element of biological activity, which can be quantified using a variety of mass spectrometric techniques to monitor incorporation of stable isotope-labelled substrates into metabolic products. This article contrasts developments in electrospray ionisation mass spectrometry (ESI-MS) for the measurement of lipid metabolism with more established gas chromatography mass spectrometry and isotope ratio mass spectrometry methodologies. ESI-MS combined with diagnostic tandem MS/MS scans permits the sensitive and specific analysis of stable isotope-labelled substrates into intact lipid molecular species without the requirement for lipid hydrolysis and derivatisation. Such dynamic lipidomic methodologies using non-toxic stable isotopes can be readily applied to quantify lipid metabolic fluxes in clinical and metabolic studies in vivo. However, a significant current limitation is the absence of appropriate software to generate kinetic models of substrate incorporation into multiple products in the time domain. Finally, we discuss the future potential of stable isotope-mass spectrometry imaging to quantify the location as well as the extent of lipid synthesis. This article is part of a Special Issue entitled: BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.     

Stable isotope ratios and forensic analysis of microorganisms

In the aftermath of the anthrax letters of 2001, researchers have been exploring various analytical signatures for the purpose of characterizing the production environment of microorganisms. One such signature is stable isotope ratios, which in heterotrophs, are a function of nutrient and water sources. Here we discuss the use of stable isotope ratios in microbial forensics, using as a database the carbon, nitrogen, oxygen, and hydrogen stable isotope ratios of 247 separate cultures of Bacillus subtilis 6051 spores produced on a total of 32 different culture media. In the context of using stable isotope ratios as a signature for sample matching, we present an analysis of variations between individual samples, between cultures produced in tandem, and between cultures produced in the same medium but at different times. Additionally, we correlate the stable isotope ratios of carbon, nitrogen, oxygen, and hydrogen for growth medium nutrients or water with those of spores and show examples of how these relationships can be used to exclude nutrient or water samples as possible growth substrates for specific cultures.     

Stable isotope analyses of feathers help identify autumn stopover sites of three long-distance migrants in northeastern Africa

The potential use of stable nitrogen (δ¹⁵N), carbon (δ¹³C) and hydrogen (δD) isotope ratios in feathers of marsh warblers Acrocephalus palustris , river warblers Locustella fluviatilis and whitethroats Sylvia communis was evaluated as a means to help identify the location and isotopic composition of autumn stopover sites in northeast Africa. Feather δD values were compared with regional precipitation δD maps averaged over autumn months. Compared with whitethroats, feather δ¹⁵N, δ¹³C, and δD values of marsh warblers and river warblers suggest the two warblers occupy and grow their feathers in geographic locations with relatively mesic environments, and with higher proportions of C3 (vs. C4) plants. However, δ¹³C values of marsh and river warblers were distinct enough to indicate use of different foodwebs. From previous studies, it is evident that during autumn stopover, river warblers moult their primaries in Ethiopia. It is likely that marsh warblers, like river warblers, stay in Ethiopia and/or in neighbouring regions. Based on feather δD values and regional δD precipitation maps, this region should lie between southeast Sudan and southwest Ethiopia. However, without additional regional isotopic maps in Africa, more precise locations of the stopover sites remain unclear. The relatively enriched δ¹⁵N and δ¹³C values of whitethroat feathers compared with the two other species, reflect the fact that whitethroats moult in relatively drier environments and/or with a lower proportion of C3 vs. C4 plants.     

Stable isotope studies in human evolution

The discipline of human evolution usually involves the evaluation of changes in gross and molecular morphology or changes in artifact assemblages. In contrast, stable isotope analysis is an indirect line of investigation. Understanding the human evolutionary sequence requires information on nutritional, biobehavioral, and general ecology. These are the kinds of information that stable isotope analysis can provide. Such studies may not identify the mechanisms for change, but their application serves to elucidate the situations under which change occurred.     

Brain imaging analysis can identify participants under regular mental training

Multivariate pattern recognition approaches have become a prominent tool in neuroimaging data analysis. These methods enable the classification of groups of participants (e.g. controls and patients) on the basis of subtly different patterns across the whole brain. This study demonstrates that these methods can be used, in combination with automated morphometric analysis of structural MRI, to determine with great accuracy whether a single subject has been engaged in regular mental training or not. The proposed approach allowed us to identify with 94.87% accuracy (p<0.001) if a given participant is a regular meditator (from a sample of 19 regular meditators and 20 non-meditators). Neuroimaging has been a relevant tool for diagnosing neurological and psychiatric impairments. This study may suggest a novel step forward: the emergence of a new field in brain imaging applications, in which participants could be identified based on their mental experience.     

Stable isotope analysis indicates trophic differences among forest floor carabids in Japan

Differences in trophic niches among carabid beetles (Coleoptera: Carabidae) co‐occurring on the forest floors of warm temperate forests in central Japan were studied using carbon (δ¹³C) and nitrogen (δ¹⁵N) stable isotope analyses. Different carabid species showed similar δ¹⁵N values, which were higher than those of their possible invertebrate prey (herbivores and detritivores) collected from the litter layer, indicating that these species were consumers in the same trophic level. In contrast, δ¹³C values differed among carabid species, indicating interspecific differences in prey animals. The variation in the δ¹³C value was larger in summer than in autumn. In summer, δ¹³C values indicated that some carabids depended highly on either grazing (low δ¹³C values) or detrital sources (high δ¹³C values) within the food chain [Chlaenius posticalis Motschulsky and Haplochlaenius costiger (Chaudoir), respectively], although other species with intermediate δ¹³C values likely depended on both. The latter group of species comprised mostly two dominant genera (Carabus and Synuchus). Although congeners might have similar feeding habits, the stable isotope ratios indicated trophic niche differences between adults of different species and between adults and larvae of the same genus.     

Stable carbon isotope composition of bone hydroxylapatite: significance in paleodietary analysis

The stable carbon isotope composition of the structural carbonate derived from animal bone hydroxylapatite (δ¹³C_B-HA) could record an animal’s diet. These records provide critical evidence for different paleontological disciplines, e.g., paleodiet analyses, and paleoclimate reconstructions. Compared to those of other body tissues, such as bone collagen or teeth enamel hydroxylapatite, δ¹³C_B-HA values record information on the whole diet of an animal in its last years. δ¹³C_B-HA can be applied to fossil animals of various body sizes. The δ¹³C analytical instruments available only require that prepared bone samples be approximately 2–5 mg for precise measurement, allowing δ¹³C_B-HA analysis to be feasible on most vertebrate fossils without destructive sampling, especially on small mammals or birds whose teeth are not large enough for sampling or are lost. Moreover, δ¹³C_B-HA can be used from different times or under less than ideal burial environments. For fossils dating back to Devonian or buried in hot and humid regions, dietary information has been completely lost in bone collagen during post-depositional processes but still remained in the δ¹³C_B-HA values because hydroxylapatite is less influenced by diagenetic effects after deposition. In addition, systematic methods such as X-ray diffraction and Fourier transform infrared spectroscopy have been developed to qualitatively or semiquantitatively assess the influence of diagenesis on bone hydroxylapatite to ensure the credibility of the δ¹³C_B-HA values. With the above merits, δ¹³C_B-HA analysis is therefore becoming an increasingly important method in paleodiet-related research. Currently, applications of the δ¹³C_B-HA method on fossil animals are primarily focused on two aspects, namely, paleodietary reconstruction of fossil animals with uncertain diets and paleoenvironmental reconstruction based on the δ¹³C_B-HA values of fossil herbivores. The published researches, combined with our new results from early birds, demonstrate the considerable significance of the δ¹³C_B-HA method in paleontological and paleoenvironmental research. Notably, the δ¹³C_B-HA-based paleodietary analysis of early vertebrates, especially the large number of small birds or mammals discovered in the past decades would be an important work in the near future.     

Stable sulfur isotope analysis of SO2 pollution impact on vegetation

Summary The ³⁴S value of SO₂ emitted by natural gas refineries is about +25, which is higher than that for non-industrial sulfur sources in our study areas. Terrestrial mosses absorb SO₂ from the atmosphere and have a ³⁴S value which is directly related to the degree of SO₂ stress to which they are subjected. The ³⁴S values for conifer needles are lower than for mosses at the same collection site, which indicates that trees obtain sulfur from both atmospheric and soil sources.Potted conifers were transferred to sites differing in their degree of SO₂ stress. This difference is reflected by the change of ³⁴S values of their needles. SO₂ absorbant pot covers, such as charcoal and moss, reduce the amount of airborne sulfur which is available to tress. Moss also may reduce SO₂ absorbed by soils in forest stands. We have used analysis of ³⁴S values to (1) help define SO₂ dispersion patterns; (2) reveal the rates at which plants accumulate this pollutant; and (3) associate suspected SO₂ injury more closely to an emission source.     

Stable isotope ecology in the Omo-Turkana Basin

Stable isotopes provide an independent assessment of paleoenvironments in the Omo-Turkana Basin. Stable isotopes track the flow of oxygen and carbon through ecosystems and accordingly are not directly related to changes in mammalian faunal composition or sedimentology. Therefore, isotope studies give insight into the paleoenvironmental conditions in which human evolutionary trends have been recorded. The development of stable isotopes as indicators of continental environmental conditions has proceeded in parallel with questions about the conditions of human environment. What was the vegetation? How hot was it? How dry? What were the diets of animals living among early humans? And most persistently, how important were "savannas" to early hominids? In this review, we take the opportunity to provide extensive background on the use of isotopes in anthropological sites. The application of stable isotope ecology to anthropological sites in the Turkana Basin has a long history, but in many ways the Omo-Turkana Basin has been a proving ground for the development of new proxy methods for understanding tropical terrestrial environments in the Neogene and Quaternary. For that reason, we also describe some of the fundamental aspects of isotope ecology that developed outside the field of paleoanthropology.     

Stable isotope ecology in the Ituri Forest

The Ituri Forest, Democratic Republic of Congo (formerly Zaire) is an example of a closed canopy forest showing extreme depletion in ¹³C. ¹³C values for plants from the canopy top, from gaps in the canopy, and from the subcanopy average –29.0±1.7, –30.4±0.9, and –34.0±1.5, respectively. The ¹³C of forest mammals show these differences, with the subcanopy browsers (okapi, dwarf antelope) having ¹³C values for tooth enamel much more negative than subcanopy frugivores who derive their food from the canopy top, and from folivores and omnivores living in gap or clearing areas. Nitrogen isotopes in plants from this ecosystem have an average ¹⁵N value of 5.4±1.8 and do not show significant differences at the 95% confidence interval between plants from the canopy top, from gaps in the canopy, and from the subcanopy. The ¹⁸O_SMOW values of surface waters in the study area are between –2.0 and –2.7. The ¹⁸O_PDB for tooth enamel ranged from –3 to +7.     

Stable isotope analysis and fin whale subpopulation structure in the eastern North Atlantic

The North Atlantic fin whale was subject to heavy exploitation in the past and, despite partial recovery, it is still considered endangered. Recent research has questioned its currently accepted subpopulation structure and migratory movements, challenging management and conservation efforts. Here we contribute to this discussion by presenting results of stable isotope analysis of fin whale bones and krill samples collected from fin whale stomachs from two exploited areas, west Iceland and northwest Spain, and comparing these results with North Atlantic isoscapes. In krill, δ¹⁵N values were highly variable and no significant differences in δ¹⁵N and δ¹³C between areas emerged. δ¹⁵N and δ¹³C values in bone were higher than in krill, due to trophic enrichment, and were not significantly different between areas. Both krill and bone results were slightly inconsistent with the local isoscapes, maybe due to seasonal variations in local productivity and in krill diet and, in the case of bone, to its capability to integrate long‐term isotopic values, derived from food consumed in distant areas. Conversely, δ¹⁸O values, which reflect the basal signal of sea water, were consistent with isoscapes and significantly higher in whales from northwestern Spain, possibly indicating migration to breeding grounds located at lower latitudes.     

Stable carbon isotope analysis reveals widespread drought stress in boreal black spruce forests

Unprecedented rates of climate warming over the past century have resulted in increased forest stress and mortality worldwide. Decreased tree growth in association with increasing temperatures is generally accepted as a signal of temperature‐induced drought stress. However, variations in tree growth alone do not reveal the physiological mechanisms behind recent changes in tree growth. Examining stable carbon isotope composition of tree rings in addition to tree growth can provide a secondary line of evidence for physiological drought stress. In this study, we examined patterns of black spruce growth and carbon isotopic composition in tree rings in response to climate warming and drying in the boreal forest of interior Alaska. We examined trees at three nested scales: landscape, toposequence, and a subsample of trees within the toposequence. At each scale, we studied the potential effects of differences in microclimate and moisture availability by sampling on northern and southern aspects. We found that black spruce radial growth responded negatively to monthly metrics of temperature at all examined scales, and we examined ?¹³C responses on a subsample of trees as representative of the wider region. The negative ?¹³C responses to temperature reveal that black spruce trees are experiencing moisture stress on both northern and southern aspects. Contrary to our expectations, ?¹³C from trees on the northern aspect exhibited the strongest drought signal. Our results highlight the prominence of drought stress in the boreal forest of interior Alaska. We conclude that if temperatures continue to warm, we can expect drought‐induced productivity declines across large regions of the boreal forest, even for trees located in cool and moist landscape positions.     

Stable isotope analysis of macroinvertebrates and their food sources in a glacier stream

1. Food sources and trophic structure of the macroinvertebrate community along a longitudinal gradient were examined in a glacier stream of the Swiss Alps (Val Roseg). Analysis of multiple stable isotopes (δ¹³C and δ¹⁵N) and measurement of C : N ratios were used to differentiate between allochthonous and autochthonous organic matter.
2. Although isotopic signatures of algae varied widely among sites and dates, it was possible to discriminate between allochthonous and autochthonous food sources using a site-specific approach.
3. Dominant food sources of herbivorous invertebrates in all main channel sites were epilithic diatoms and the filamentous gold alga Hydrurus foetidus . Allochthonous organic matter was of some importance only in a groundwater-fed stream close to the floodplain margin.
4. Seasonal changes in the δ¹³C signature of the macroinvertebrates corresponded with seasonal changes in δ¹³C of the gold alga H. foetidus . This indicated that the energy base remains autochthonous throughout the year.
5. Because of limited food sources, feeding plasticity of the invertebrate community was high. Both grazers and shredders fed predominantly on algae, whereas gatherer-collectors seemed to be omnivorous.
6. The overall enrichment of δ¹⁵N was 2.25‰ ( r ²=0.99) per trophic level. On a gradient from the glacier site to a downstream forested site trophic enrichment was constant but variation in δ¹⁵N within trophic levels decreased.     

Amplification and analysis of human DNA present in mosquito bloodmeals

Abstract. DNA fingerprinting should permit the identification of individual human hosts of haematophagous arthropods, providing epidemi-ologically useful information, for example, the biting rates on different people and the impact of insecticide-impregnated bednets.
Investigations reported here demonstrate that it is possible to extract, amplify and fingerprint human DNA from the bloodmeals of individual female Anopheles gambiae mosquitoes kept at 24^oC for up to 10–15 h post-ingestion.