, Volume 191, Issue 4, pp 745–755 | Cite as

Does lipid-correction introduce biases into isotopic mixing models? Implications for diet reconstruction studies

  • Martin C. ArosteguiEmail author
  • Daniel E. Schindler
  • Gordon W. Holtgrieve
Concepts, Reviews and Syntheses


Carbon isotopes are commonly used in trophic ecology to estimate consumer diet composition. This estimation is complicated by the fact that lipids exhibit a more depleted carbon signature (δ13C) than other macromolecules, and are often found at different concentrations among individual organisms. Some researchers argue that lipids bias diet reconstructions using stable isotopes and should be accounted for prior to analysis in food web mixing models, whereas others contend that removing lipids may result in erroneous interpretations of the trophic interactions under study. To highlight this disagreement on best practices for applying δ13C in food web studies, we sampled the recent literature to determine the frequency and method of lipid-correction. We then quantified the potential magnitude and source of bias in mixing model results from a theoretical example and case study of diet reconstruction. The literature was split nearly evenly as to whether lipid-correction was applied to δ13C data in mixing model estimates of diet composition. Comparative mixing model scenarios demonstrated that lipid-correction can substantially alter the estimated diet composition and interpretation of consumer foraging habits. Given the lack of consensus on whether or not to lipid-correct prey and/or consumers, and the associated variation in mixing model results, we call for the establishment of a unified framework that will guide diet reconstruction in stable isotope ecology. Uncertainty in the prevalence of direct routing versus de novo synthesis of lipids across ecosystems, taxa, and trophic levels must be resolved to better guide treatment of lipids in isotope studies using carbon.


Carbon Diet Lipid Lipid-correction Mixing model Stable isotope 



Funding for this project was provided by the Achievement Rewards for College Scientists (ARCS) Foundation Seattle Chapter via the Barton family, Richard and Lois Worthington Endowment, H. Mason Keeler Endowment, Clarence H. Campbell Endowed Lauren Donaldson Scholarship in Ocean and Fishery Sciences, Richard T. Whiteleather Fisheries B.S. 1935 Endowed Scholarship, and Floyd E. Ellis Memorial Scholarship. We thank Thomas Quinn, Michael Brett, Joel Trexler, and three anonymous reviewers for critical feedback on the manuscript.

Author contribution statement

MCA, DES, and GWH designed the study. MCA conducted the literature review, mixing model comparisons and wrote the first draft, after which DES and GWH contributed to the editing and refinement of the manuscript.

Supplementary material

442_2019_4525_MOESM1_ESM.docx (85 kb)
Supplementary material 1 (DOCX 84 kb)
442_2019_4525_MOESM2_ESM.docx (77 kb)
Supplementary material 2 (DOCX 77 kb)


  1. Ambrose SH, Norr L (1993) Carbon isotopic evidence for routing of dietary protein to bone collagen, and whole diet to bone apatite carbonate: purified diet growth experiments. In: Lambert JB, Grupe G (eds) Molecular archaeology of prehistoric human bone. Springer, Berlin, pp 1–37Google Scholar
  2. Arts MT, Achman RG, Holub BJ (2001) “Essential fatty acids” in aquatic ecosystems: a crucial link between diet and human health and evolution. Can J Fish Aquat Sci 58:122–137Google Scholar
  3. Babar A, Jayawant MS, Pawar SP (2017) Nutritional profile of the freshwater edible bivalve Lamellidens corrianus (Lea 1834) and its relation to water quality in the Bhatsa River, India. Asian Fish Sci 30:52–69Google Scholar
  4. Beukema JJ (1997) Caloric values of marine invertebrates with an emphasis on the soft parts of marine bivalves. Oceanogr Mar Biol 35:387–414Google Scholar
  5. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917Google Scholar
  6. Boecklen WJ, Yarnes CT, Cook BA, James AC (2011) On the use of stable isotopes in trophic ecology. Annu Rev Ecol Evol Syst 42:411–440Google Scholar
  7. Bond AL, Diamond AW (2011) Recent Bayesian stable-isotope mixing models are highly sensitive to variation in discrimination factors. Ecol Appl 21:1017–1023PubMedGoogle Scholar
  8. Brett MT (2014) Resource polygon geometry predicts Bayesian stable isotope mixing model bias. Mar Ecol Prog Ser 514:1–12Google Scholar
  9. Brett MT, Müller-Navarra DC (1997) The role of highly unsaturated fatty acids in aquatic foodweb processes. Freshw Biol 38:483–499Google Scholar
  10. Brett MT, Müller-Navarra DC, Ballantyne AP, Ravet JL, Goldman CR (2006) Daphnia fatty acid composition reflects that of their diet. Limnol Oceanogr 51:2428–2437Google Scholar
  11. Buchheister A, Latour RJ (2010) Turnover and fractionation of carbon and nitrogen stable isotopes in tissues of a migratory coastal predator, summer flounder (Paralichthys dentatus). Can J Fish Aquat Sci 67:445–461Google Scholar
  12. Cherry SG, Derocher AE, Hobson KA, Stirling I, Thiemann GW (2011) Quantifying dietary pathways of proteins and lipids to tissues of a marine predator. J Appl Ecol 48:373–381Google Scholar
  13. Cummings BM, Schindler DE (2013) Depth variation in isotopic composition of benthic resources and assessment of sculpin feeding patterns in an oligotrophic Alaskan lake. Aquat Ecol 47:403–414Google Scholar
  14. DeNiro MJ, Epstein S (1977) Mechanism of carbon isotope fractionation associated with lipid synthesis. Science 197:261–263PubMedGoogle Scholar
  15. DeNiro MJ, Epstein S (1978) Influence of diet on the distribution of carbon isotopes in animals. Geochim Cosmochim Acta 42:495–506Google Scholar
  16. Erhardt EB, Wolf BO, Ben-David M, Bedrick EJ (2014) Stable isotope sourcing using sampling. Open J Ecol 4:289–298Google Scholar
  17. Felip O, Ibarz A, Fernández-Borrás J, Beltrán M, Martín-Pérez M, Planas JV, Blasco J (2012) Tracing metabolic routes of dietary carbohydrate and protein in rainbow trout (Oncorhynchus mykiss) using stable isotopes ([13C] starch and [15N] protein): effects of gelatinization of starches and sustained swimming. Br J Nutr 107:834–844PubMedGoogle Scholar
  18. Fernandes R, Millard AR, Brabec M, Nadeau M-J, Grootes P (2014) Food reconstruction using isotopic transferred signals (Fruits): a Bayesian model for diet reconstruction. PLoS One 9:e87436PubMedPubMedCentralGoogle Scholar
  19. Fernandes R, Larsen T, Knipper C, Feng F, Wang Y (2017) a database of isotopic data for ecology, archaeology, and environmental science. Accessed 12 July 2018.
  20. Fry B (2006) Stable isotope ecology. Springer, New YorkGoogle Scholar
  21. Fry B, Baltz DM, Benfield MC, Fleeger JW, Gace A, Haas HL, Quinones-Rivera ZJ (2003) Stable isotope indicators of movement and residency for brown shrimp (Farfantepenaeus aztecus) in coastal Louisiana marshscapes. Estuaries 26:82–97Google Scholar
  22. Galloway AWE, Eisenlord ME, Dethier MN, Holtgrieve GW, Brett MT (2014) Quantitative estimates of isopod resource utilization using a Bayesian fatty acid mixing model. Mar Ecol Prog Ser 507:219–232Google Scholar
  23. Garcés R, Mancha M (1993) One-step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Anal Biochem 211:139–143PubMedGoogle Scholar
  24. Gaye-Siessegger J, Focken U, Abel H, Becker K (2004) Dietary lipid content influences the activity of lipogenic enzymes in the liver and on whole body δ13C values of Nile tilapia, Oreochromis niloticus (L.). Isot Environ Health Stud 40:181–190Google Scholar
  25. Gelman A, Carlin JB, Stern HS, Dunson DB, Vehtari A, Rubin DB (2013) Bayesian data analysis, 3rd edn. CRC Press, Boca RatonGoogle Scholar
  26. Gende SM, Quinn TP, Willson MF (2001) Consumption choice by bears feeding on salmon. Oecologia 127:372–382PubMedGoogle Scholar
  27. Gende SM, Quinn TP, Hilborn R, Hendry AP, Dickerson B (2004) Brown bears selectively kill salmon with higher energy content but only in habitats that facilitate choice. Oikos 104:518–528Google Scholar
  28. Goetz F, Rosauer D, Sitar S, Goetz G, Simchick C, Roberts S, Johnson R, Murphy C, Bronte CR, Mackenzie S (2010) A genetic basis for the phenotypic differentiation between siscowet and lean lake trout (Salvelinus namaycush). Mol Ecol 19:176–196PubMedGoogle Scholar
  29. Goulden CE, Place AR (1990) Fatty acid synthesis and accumulation rates in daphniids. J Exp Zool 256:168–178Google Scholar
  30. Haramis GM, Nichols JD, Pollock KH, Hines JE (1986) The relationship between body mass and survival of wintering canvasbacks. Auk 103:506–514Google Scholar
  31. Harvey CJ, Hanson PC, Essington TE, Brown PB, Kitchell JF (2002) Using bioenergetics models to predict stable isotope ratios in fishes. Can J Fish Aquat Sci 59:115–124Google Scholar
  32. Hoffman JC, Sierszen ME, Cotter AM (2015) Fish tissue lipid-C: N relationships for correcting values and estimating lipid content in aquatic food-web studies. Rapid Commun Mass Spectrom 29:2069–2077PubMedGoogle Scholar
  33. Hopkins CA (1950) Studies on cestode metabolism. I. Glycogen metabolism in Schistocephalus solidus in vivo. J Parasitol 26:384–390Google Scholar
  34. Janjua MY, Gerdeaux D (2011) Evaluation of food web and fish dietary niches in oligotrophic Lake Annecy by gut content and stable isotope analysis. Lake Reserv Manag 27:115–127Google Scholar
  35. Kelly LJ, Martínez del Rio C (2010) The fate of carbon in growing fish: an experimental study of isotopic routing. Physiol Biochem Zool 83:473–480PubMedGoogle Scholar
  36. Kilborn L, Macleod J (1920) Observation on the glycogen content of certain invertebrates and fishes. Exp Physiol 12:317–330Google Scholar
  37. Kiljunen M, Grey J, Sinisalo T, Harrod C, Immonen H, Jones RI (2006) A revised model for lipid-normalizing values from aquatic organisms, with implications for isotope mixing models. J Appl Ecol 43:1213–1222Google Scholar
  38. Layman CA, Araujo MS, Boucek R, Hammerschlag-Peyer CM, Harrison E, Jud ZR, Matich P, Rosenblatt AE, Vaudo JJ, Yeager LA, Post DM, Bearhop S (2012) Applying stable isotopes to examine food-web structure: an overview of analytical tools. Biol Rev 87:545–562PubMedGoogle Scholar
  39. Lincoln AE, Quinn TP (2018) Optimal foraging or surplus killing: selective consumption and discarding of salmon by brown bears. Behav Ecol 30:202–212Google Scholar
  40. Logan JM, Jardine TD, Miller TJ, Bunn SE, Cunjak RA, Lutcavage ME (2008) Lipid corrections in carbon and nitrogen stable isotope analyses: comparison of chemical extraction and modeling methods. J Anim Ecol 77:838–846PubMedGoogle Scholar
  41. Martínez del Rio C, Wolf N, Carleton S, Gannes LZ (2009) Isotopic ecology ten years after a call for more laboratory experiments. Biol Rev 84:91–111Google Scholar
  42. Mayntz D, Raubenheimer D, Salomon M, Toft S, Simpson SJ (2005) Nutrient-specific foraging in invertebrate predators. Science 307:111–113PubMedGoogle Scholar
  43. Mayntz D, Nielsen VH, Sørensen A, Toft S, Raubenheimer D, Hejlesen C, Simpson SJ (2009) Balancing of protein and lipid intake by a mammalian carnivore, the mink, Mustela vison. Anim Behav 77:349–355Google Scholar
  44. McConnaughey T, McRoy CP (1979) Food-web structure and the fractionation of carbon isotopes in the Bering Sea. Mar Biol 53:257–262Google Scholar
  45. Mohan JA, Smith SD, Connelly TL, Attwood ET, McClelland JW, Herzka SZ, Walther BD (2016) Tissue-specific isotope turnover and discrimination factors are affected by diet quality and lipid content in an omnivorous consumer. J Exp Mar Biol Ecol 479:35–45Google Scholar
  46. Moore JW, Semmens BX (2008) Incorporating uncertainty and prior information into stable isotope mixing models. Ecol Lett 1:470–480Google Scholar
  47. Morrison RIG, Davidson NC, Wilson JR (2007) Survival of the fattest: body stores on migration and survival in red knots Calidris canutus islandica. J Avian Biol 38:479–487Google Scholar
  48. Newsome SD, Wolf N, Peters J, Fogel ML (2014) Amino acid analysis shows flexibility in the routing of dietary protein and lipids to the tissue of an omnivore. Integr Comp Biol 54:890–902PubMedGoogle Scholar
  49. Nithirojpakdee P, Beamish FWH, Boonphakdee T (2014) Diet diversity among five co-existing fish species in a tropical river: integration of dietary and stable isotope data. Limnology 15:99–107Google Scholar
  50. O’Neill SM, Ylitalo GM, West JE (2014) Energy content of Pacific salmon as prey of northern and southern resident killer whales. Endanger Species Res 25:265–281Google Scholar
  51. Park R, Epstein S (1961) Metabolic fractionation of C13 and C12 in plants. Plant Physiol 36:133–138PubMedPubMedCentralGoogle Scholar
  52. Parnell AC, Inger R, Bearhop S, Jackson AL (2010) Source partitioning using stable isotopes: coping with too much variation. PLoS One 5:e9672PubMedPubMedCentralGoogle Scholar
  53. Parrish FA, Martinelli-Liedtke TL (1999) Some preliminary findings on the nutritional status of the Hawaiian spiny lobster (Panulirus marginatus). Pac Sci 53:361–366Google Scholar
  54. Patterson HK, Carmichael RH (2016) The effect of lipid extraction on carbon and nitrogen stale isotope ratios in oyster tissues: implications for glycogen-rich species. Rapid Commun Mass Spectrom 30:2594–2600PubMedGoogle Scholar
  55. Pauli JN, Newsome SD, Cook JA, Harrod C, Steffan SA, Baker CJO, Ben-David M, Bloom D, Bowen GJ, Cerling TE, Cicero C, Cook C, Dohm M, Dharampal PS, Graves G, Gropp R, Hobson KA, Jordan C, MacFadden B, Birch SP, Poelen J, Ratnasignham S, Russell L, Stricker CA, Uhen MD, Yarnes CT, Hayden B (2017) Why we need a centralized repository for isotopic data. Proc Natl Acad Sci USA 114:2997–3001PubMedGoogle Scholar
  56. Pecquerie L, Nisbet RM, Fablet R, Lorrain A, Koojiman SA (2010) The impact of metabolism on stable isotope dynamics: a theoretical framework. Phil Trans R Soc B 365:3455–3468PubMedGoogle Scholar
  57. Peterson BJ, Fry B (1987) Stable isotopes in ecosystem studies. Ann Rev Ecol Syst 18:293–320Google Scholar
  58. Phillips DL, Gregg JW (2003) Source partitioning using stable isotopes: coping with too many sources. Oecologia 136:261–269PubMedGoogle Scholar
  59. Phillips DL, Inger R, Bearhop S, Jackson AL, Moore JW, Parnell AC, Semmens BX, Ward EJ (2014) Best practices for use of stable isotope mixing models in food-web studies. Can J Zoo 92:823–835Google Scholar
  60. Podlesak DW, McWilliams SR (2007) Metabolic routing of dietary nutrients in birds: effects of dietary lipid concentration on δ13C of depot fat and its ecological implications. Auk 124:916–925Google Scholar
  61. Post DM (2002) Using stable isotopes to estimate trophic position: models, methods, and assumptions. Ecology 83:703–718Google Scholar
  62. Post DM, Layman CA, Arrington DA, Takimoto G, Quattrochi J, Montaña CG (2007) Getting to the fat of the matter: models, methods and assumptions for dealing with lipids in stable isotope analyses. Oecologia 152:179–189PubMedGoogle Scholar
  63. Rau GH (1980) Carbon-13/carbon-12 variation in subalpine lake aquatic insects: food source implications. Can J Fish Aquat Sci 37:742–746Google Scholar
  64. Reimchen TE (2000) Some ecological and evolutionary aspects of bear-salmon interactions in coastal British Columbia. Can J Zool 78:448–457Google Scholar
  65. Reitan KI, Rainuzzo JR, Olsen Y (1994) Effect of nutrient limitation on fatty acid and lipid content of marine microalgae. J Phycol 30:972–979Google Scholar
  66. Rowe DK, Thorpe JE, Shanks AM (1991) Role of fat stores in the maturation of male Atlantic salmon (Salmo salar) parr. Can J Fish Aquat Sci 48:405–413Google Scholar
  67. Ryan C, McHugh B, Trueman CN, Harrod C, Berrow SD, O’Connor I (2012) Accounting for the effects of lipids in stable isotope (δ13C and δ15N values) analysis of skin and blubber of balaenopterid whales. Rapid Commun Mass Spectrom 26:2745–2754PubMedGoogle Scholar
  68. Sanderson BL, Tran CD, Coe HJ, Pelekis V, Steel A, Reichert WL (2009) Nonlethal sampling of fish caudal fins yields valuable stable isotope data for threatened and endangered fishes. Trans Am Fish Soc 138:1166–1177Google Scholar
  69. Shine R, Madsen T (1997) Prey abundance and predator reproduction: rats and pythons on a tropical Australian floodplain. Ecology 78:1078–1086Google Scholar
  70. Smith RJ, Moore FR (2003) Arrival fat and reproductive performance in a long-distance passerine migrant. Oecologia 134:325–331PubMedGoogle Scholar
  71. Stock BC, Semmens BX (2013) MixSIAR GUI User Manual. Version 3.1. Accessed Dec 2018
  72. Stock BC, Semmens BX (2016) Unifying error structures in commonly used biotracer mixing models. Ecology 97:2562–2569PubMedGoogle Scholar
  73. Stock BC, Jackson AL, Ward EJ, Parnell AC, Phillips DL, Semmens BX (2018) Analyzing mixing systems using a new generation of Bayesian tracer mixing models. PeerJ 6:e5096PubMedPubMedCentralGoogle Scholar
  74. Sweeting CJ, Polunin NVC, Jennings S (2006) Effects of chemical lipid extraction and arithmetic lipid correction on stable isotope ratios of fish tissues. Rapid Commun Mass Spectrom 20:595–601PubMedGoogle Scholar
  75. Taipale SJ, Kainz MJ, Brett MT (2011) Diet-switching experiments show rapid accumulation and preferential retention of highly unsaturated fatty acids in Daphnia. Oikos 120:1674–1982Google Scholar
  76. Tarroux A, Ehrich D, Lecomte N, Jardine TD, Bêty J, Berteaux D (2010) Sensitivity of stable isotope mixing models to variation in isotopic ratios: evaluating consequences of lipid extraction. Methods Ecol Evol 1:231–241Google Scholar
  77. Tocher DR (2003) Metabolism and functions of lipids and fatty acids in teleost fish. Rev Fish Sci 11:107–184Google Scholar
  78. van der Merwe NJ (1982) Carbon isotopes, photosynthesis, and archaeology. Am Sci 70:596–606Google Scholar
  79. Wessels FJ, Hahn DA (2010) Carbon 13 discrimination during lipid biosynthesis varies with dietary concentration of stable isotopes: implications for stable isotope analyses. Funct Ecol 24:1017–1022Google Scholar
  80. Wolf N, Newsome SD, Peters J, Fogel ML (2015) Variability in the routing of dietary proteins and lipids to consumer tissues influences tissue-specific isotopic discrimination. Rapid Commun Mass Spectrom 29:1448–1456PubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Aquatic and Fishery SciencesUniversity of WashingtonSeattleUSA
  2. 2.Applied Physics Laboratory, Air-Sea Interaction & Remote Sensing DepartmentUniversity of WashingtonSeattleUSA

Personalised recommendations