Is acidification of samples for isotopic analysis of carbon and nitrogen necessary for shoreline marine species?
Larissa M. Pires-Teixeira A C , Vinicius Neres-Lima B and Joel C. Creed BA Programa de Pós-Graduação em Ecologia e Evolução, Universidade do Estado do Rio de Janeiro, Rua Francisco Xavier 524, PHLC, Sala 220, 20559-900, Rio de Janeiro, RJ, Brazil.
B Departamento de Ecologia, IBRAG, Universidade do Estado do Rio de Janeiro, Rua Francisco Xavier 524, PHLC, Sala 220, 20559-900, Rio de Janeiro, RJ, Brazil.
C Corresponding author. Email: larissamarques@ymail.com
Marine and Freshwater Research 72(2) 256-262 https://doi.org/10.1071/MF19227
Submitted: 28 June 2019 Accepted: 1 June 2020 Published: 17 July 2020
Abstract
The acidification of samples for the simultaneous measurement of stable carbon (δ13C) and nitrogen (δ15N) isotopes represents an important methodological question still not clarified because the removal of calcium carbonate (CaCO3) from samples may affect the outcome of the stable-isotope analysis. We investigated the effect of acidification on samples of 25 different taxa of benthic marine organisms from a rocky shore, to determine whether acidification affects the isotopic values of δ13C and δ15N. After washing with distilled water and drying, each sample was divided into two parts; one part was acidified by adding 1 M HCl with a pipette, until no more CO2 was released, whereas the other part was retained as a non-acidified sample. Organisms with little CaCO3 showed no difference in carbon isotopic signatures after acidification, from those in the non-acidified part. Some organisms with a high CaCO3 content did show differences in carbon isotopic signatures after acidification. The nitrogen isotopic signature presented a significant difference; however, to what extent this difference is biologically important is discussed. Avoiding sampling parts containing a high concentration of carbonate is an efficient way to circumvent this problem. Acidification of samples as a pre-treatment for isotopic analysis is not useful for marine organisms without calcareous structures and is necessary only when the sampled parts contain significant amounts of carbonate.
Additional keywords: decarbonation, rocky shore, sample treatment, stable isotope.
References
Bilan, M. I., and Usov, A. I. (2001). Polysaccharides of calcareous algae and their effect on the calcification process. Russian Journal of Bioorganic Chemistry 27, 2–16.| Polysaccharides of calcareous algae and their effect on the calcification process.Crossref | GoogleScholarGoogle Scholar |
Bosley, K. L., and Wainright, S. C. (1999). Effects of preservatives and acidification on the stable isotope ratios (15N : 14N, 13C : 12C) of two species of marine animals. Canadian Journal of Fisheries and Aquatic Sciences 56, 2181–2185.
| Effects of preservatives and acidification on the stable isotope ratios (15N : 14N, 13C : 12C) of two species of marine animals.Crossref | GoogleScholarGoogle Scholar |
Boßelmann, F., Romano, P., Fabritius, H., Raabe, D., and Epple, M. (2007). The composition of the exoskeleton of two crustacea: the American lobster Homarus americanus and the edible crab Cancer pagurus. Thermochimica Acta 463, 65–68.
| The composition of the exoskeleton of two crustacea: the American lobster Homarus americanus and the edible crab Cancer pagurus.Crossref | GoogleScholarGoogle Scholar |
Brusca, R. C., and Brusca, G. J. (2007). ‘Invertebrados.’ 2nd Edn. (Guanabara Koogan: Rio de Janeiro, Brazil.)
Bunn, S. E., Loneragan, N. R., and Kempster, M. A. (1995). Effects of acid washing on stable isotope ratios of C and N in penaeid shrimp and seagrass: implications for food‐web studies using multiple stable isotopes. Limnology and Oceanography 40, 622–625.
| Effects of acid washing on stable isotope ratios of C and N in penaeid shrimp and seagrass: implications for food‐web studies using multiple stable isotopes.Crossref | GoogleScholarGoogle Scholar |
Carabel, S., Godínez-Domínguez, E., Verísimo, P., Fernández, L., and Freire, J. (2006). An assessment of sample processing methods for stable isotope analyses of marine food webs. Journal of Experimental Marine Biology and Ecology 336, 254–261.
| An assessment of sample processing methods for stable isotope analyses of marine food webs.Crossref | GoogleScholarGoogle Scholar |
Carlier, A., Riera, P., Amouroux, J. M., Bodiou, J. Y., and Grémare, A. (2007). Benthic trophic network in the Bay of Banyuls-sur-Mer (northwest Mediterranean, France): an assessment based on stable carbon and nitrogen isotopes analysis. Estuarine, Coastal and Shelf Science 72, 1–15.
| Benthic trophic network in the Bay of Banyuls-sur-Mer (northwest Mediterranean, France): an assessment based on stable carbon and nitrogen isotopes analysis.Crossref | GoogleScholarGoogle Scholar |
Chanton, J. P., and Lewis, F. G. (1999). Plankton and dissolved inorganic carbon isotopic composition in a river-dominated estuary: Apalachicola Bay, Florida. Estuaries 22, 575–583.
| Plankton and dissolved inorganic carbon isotopic composition in a river-dominated estuary: Apalachicola Bay, Florida.Crossref | GoogleScholarGoogle Scholar |
Corbisier, T. N., Soares, L. S. H., Petti, M. A. V., Muto, E. Y., Silva, M. H. C., McClelland, J., and Valiela, I. (2006). Use of isotopic signatures to assess the food web in a tropical shallow marine ecosystem of southeastern Brazil. Aquatic Ecology 40, 381–390.
| Use of isotopic signatures to assess the food web in a tropical shallow marine ecosystem of southeastern Brazil.Crossref | GoogleScholarGoogle Scholar |
DeNiro, M. J., and Epstein, S. (1978). Influence of diet on the distribution of carbon isotopes in animals. Geochimica et Cosmochimica Acta 42, 495–506.
| Influence of diet on the distribution of carbon isotopes in animals.Crossref | GoogleScholarGoogle Scholar |
Fry, B. (1988). Food web structure on Georges Bank from stable C, N, and S isotopic compositions. Limnology and Oceanography 33, 1182–1190.
| Food web structure on Georges Bank from stable C, N, and S isotopic compositions.Crossref | GoogleScholarGoogle Scholar |
Fry, B. (2006). ‘Stable Isotope Ecology.’ (Springer Publishing: New York, NY, USA.)
Goering, J., Alexander, V., and Haubenstock, N. (1990). Seasonal variability of stable carbon and nitrogen isotope ratios of organisms in a North Pacific bay. Estuarine, Coastal and Shelf Science 30, 239–260.
| Seasonal variability of stable carbon and nitrogen isotope ratios of organisms in a North Pacific bay.Crossref | GoogleScholarGoogle Scholar |
Goreau, T. F. (1963). Calcium carbonate deposition by coralline algae and corals in relation to their roles as reef-builders. Annals of the New York Academy of Sciences 109, 127–167.
| Calcium carbonate deposition by coralline algae and corals in relation to their roles as reef-builders.Crossref | GoogleScholarGoogle Scholar | 13949254PubMed |
Jacob, U., Mintenbeck, K., Brey, T., Knust, R., and Beyer, K. (2005). Stable isotope food web studies: a case for standardized sample treatment. Marine Ecology Progress Series 287, 251–253.
| Stable isotope food web studies: a case for standardized sample treatment.Crossref | GoogleScholarGoogle Scholar |
Jaschinski, S., Hansen, T., and Sommer, U. (2008). Effects of acidification in multiple stable isotope analyses. Limnology and Oceanography, Methods 6, 12–15.
| Effects of acidification in multiple stable isotope analyses.Crossref | GoogleScholarGoogle Scholar |
Jones, G. E., and Starkey, R. L. (1957). Fractionation of stable isotopes of sulfur by microorganisms and their role in deposition of native sulfur. Applied Microbiology 5, 111–118.
| Fractionation of stable isotopes of sulfur by microorganisms and their role in deposition of native sulfur.Crossref | GoogleScholarGoogle Scholar | 13425528PubMed |
Layman, C. A., Arrington, D. A., Montaña, C. G., and Post, D. M. (2007). Can stable isotope ratios provide for community-wide measures of trophic structure? Ecology 88, 42–48.
| Can stable isotope ratios provide for community-wide measures of trophic structure?Crossref | GoogleScholarGoogle Scholar | 17489452PubMed |
Lewis, W. M., Hamilton, S. K., Rodríguez, M. A., Saunders, J. F., and Lasi, M. A. (2001). Foodweb analysis of the Orinoco floodplain based on production estimates and stable isotope data. Journal of the North American Benthological Society 20, 241–254.
| Foodweb analysis of the Orinoco floodplain based on production estimates and stable isotope data.Crossref | GoogleScholarGoogle Scholar |
Longhurst, A. R., and Pauly, D. (1987). ‘Ecology of Tropical Oceans.’ (Academic Press: San Diego, CA, USA.)
Marcus, L., Virtue, P., Nichols, P. D., Meekan, M. G., and Pethybridge, H. (2017). Effects of sample treatment on the analysis of stable isotopes of carbon and nitrogen in zooplankton, micronekton and a filter-feeding shark. Marine Biology 164, 124.
| Effects of sample treatment on the analysis of stable isotopes of carbon and nitrogen in zooplankton, micronekton and a filter-feeding shark.Crossref | GoogleScholarGoogle Scholar |
Mateo, M. A., Serrano, O., Serrano, L., and Michener, R. H. (2008). Effects of sample preparation on stable isotope ratios of carbon and nitrogen in marine invertebrates: implications for food web studies using stable isotopes. Oecologia 157, 105–115.
| Effects of sample preparation on stable isotope ratios of carbon and nitrogen in marine invertebrates: implications for food web studies using stable isotopes.Crossref | GoogleScholarGoogle Scholar | 18465146PubMed |
McCutchan, J. H., Lewis, W. M., Kendall, C., and Mcgrath, C. C. (2003). Variation in trophic shift for stable isotope ratios of carbon, nitrogen, and sulfur. Oikos 102, 378–390.
| Variation in trophic shift for stable isotope ratios of carbon, nitrogen, and sulfur.Crossref | GoogleScholarGoogle Scholar |
McIntyre, A. D., and Eleftheriou, A. (2005). ‘Methods for the Study of Marine Benthos.’ (Blackwell Science Publications: Oxford, UK.)
Michener, R., and Lajtha, K. (2008). ‘Stable Isotopes in Ecology and Environmental Science.’ 2nd Edn. (Blackwell Publishing: Melbourne, Vic, Australia.)
Minagawa, M., and Wada, E. (1984). Stepwise enrichment of 15N along food chains: further evidence and the relation between δ15N and animal age. Geochimica et Cosmochimica Acta 48, 1135–1140.
| Stepwise enrichment of 15N along food chains: further evidence and the relation between δ15N and animal age.Crossref | GoogleScholarGoogle Scholar |
Neres-Lima, V., Brito, E. F., Krsulović, F. A., Detweiler, A. M., Hershey, A. E., and Moulton, T. P. (2016). High importance of autochthonous basal food source for the food web of a Brazilian tropical stream regardless of shading. International Review of Hydrobiology 101, 132–142.
| High importance of autochthonous basal food source for the food web of a Brazilian tropical stream regardless of shading.Crossref | GoogleScholarGoogle Scholar |
Neres-Lima, V., Machado-Silva, F., Baptista, D. F., Oliveira, R., Andrade, P. M., Oliveira, A. F., Sasada-Sato, C. Y., Silva-Junior, E. F., Feijó-Lima, R., Angelini, R., Camargo, P. B., and Moulton, T. P. (2017). Allochthonous and autochthonous carbon flows in food webs of tropical forest streams. Freshwater Biology 62, 1012–1023.
| Allochthonous and autochthonous carbon flows in food webs of tropical forest streams.Crossref | GoogleScholarGoogle Scholar |
Newsome, S. D., Martinez del Rio, C., Bearhop, S., and Phillips, D. L. (2007). A niche for isotopic ecology. Frontiers in Ecology and the Environment 5, 429–436.
| A niche for isotopic ecology.Crossref | GoogleScholarGoogle Scholar |
Ng, J. S. S., Wai, T. C., and Williams, G. A. (2007). The effects of acidification on the stable isotope signatures of marine algae and molluscs. Marine Chemistry 103, 97–102.
| The effects of acidification on the stable isotope signatures of marine algae and molluscs.Crossref | GoogleScholarGoogle Scholar |
Pereira, R. C., and Soares-Gomes, A. (2002). ‘Biologia Marinha.’ 1st Edn. (Interciência: Rio de Janeiro, Brazil.)
Peterson, B. J. (1999). Stables isotopes as tracers of organic matter input and transfer in benthic food webs: a review. Acta Oecologica 20, 479–487.
| Stables isotopes as tracers of organic matter input and transfer in benthic food webs: a review.Crossref | GoogleScholarGoogle Scholar |
Pinnegar, J. K., and Polunin, N. V. C. (1999). Differential fractionation of δ13C and δ15N among fish tissues: implications for the study of trophic interactions. Functional Ecology 13, 225–231.
| Differential fractionation of δ13C and δ15N among fish tissues: implications for the study of trophic interactions.Crossref | GoogleScholarGoogle Scholar |
Riebesell, U., Zondervan, I., Rost, B., Tortell, P. D., Zeebe, R. E., and Morel, F. M. (2000). Reduced calcification of marine plankton in response to increased atmospheric CO2. Nature 407, 364–367.
| Reduced calcification of marine plankton in response to increased atmospheric CO2.Crossref | GoogleScholarGoogle Scholar | 11014189PubMed |
Salgado, L. T., Amado Filho, G. M., Fernandez, M. S., Arias, J. L., and Farina, M. (2011). The effect of alginates, fucans and phenolic substances from the brown seaweed Padina gymnospora in calcium carbonate mineralization in vitro. Journal of Crystal Growth 321, 65–71.
| The effect of alginates, fucans and phenolic substances from the brown seaweed Padina gymnospora in calcium carbonate mineralization in vitro.Crossref | GoogleScholarGoogle Scholar |
Serrano, O., Serrano, L., Mateo, M. A., Colombini, I., Chelazzi, L., Gagnarli, E., and Fallaci, M. (2008). Acid washing effect on elemental and isotopic composition of whole beach arthropods: implications for food web studies using stable isotopes. Acta Oecologica 34, 89–96.
| Acid washing effect on elemental and isotopic composition of whole beach arthropods: implications for food web studies using stable isotopes.Crossref | GoogleScholarGoogle Scholar |
Søreide, J. E., Tamelander, T., Hop, H., Hobson, K. A., and Johansen, I. (2006). Sample preparation effects on stable C and N isotope values: a comparison of methods in Arctic marine food web studies. Marine Ecology Progress Series 328, 17–28.
| Sample preparation effects on stable C and N isotope values: a comparison of methods in Arctic marine food web studies.Crossref | GoogleScholarGoogle Scholar |
Vafeiadou, A.-M., Adão, H., De Troch, M., and Moens, T. (2013). Sample acidification effects on carbon and nitrogen stable isotope ratios of macrofauna from a Zostera noltii bed. Marine and Freshwater Research 64, 741–745.
| Sample acidification effects on carbon and nitrogen stable isotope ratios of macrofauna from a Zostera noltii bed.Crossref | GoogleScholarGoogle Scholar |
Vanderklift, M. A., and Ponsard, S. (2003). Sources of variation in consumer-diet δ15N enrichment: a meta-analysis. Oecologia 136, 169–182.
| Sources of variation in consumer-diet δ15N enrichment: a meta-analysis.Crossref | GoogleScholarGoogle Scholar | 12802678PubMed |