Respiration: comparison of the Winkler technique, O2 electrodes, O2 optodes and the respiratory electron transport system assay
- 203 Downloads
Aerobic respiration is a biological energy generation process that consumes organic carbon and oxygen. In the ocean, the balance between photosynthesis and respiration is recognized as critical to understanding the ocean’s impact on the hydrospheric and atmospheric CO2. Techniques to determine respiration can be based on inorganic chemistry, electrochemistry, photochemistry, and enzymology. Here, for method comparison, physiological respiration was simultaneously measured by the Winkler method (W), O2 electrodes (E), and O2 optodes (O). These techniques detected respiratory O2 consumption (R), in situ, in dark incubation chambers. Respiratory electron transport system activity measurements detected potential respiration (Ф), biochemically. Leptomysis lingvura, a marine mysid, and Ulva rigida, a species of green algal sea lettuce, were the two organisms tested. Physiological respiration results from each technique were not statistically significantly different (multiple paired Student’s t tests, p value > 0.05) and were inside the range of similar published measurements. The mean dry-mass-specific respiration in L. lingvura and U. rigida was 0.147 ± 0.037 and 0.023 ± 0.008 µmol O2 h−1 (mg dry mass)−1, n = 9, respectively. The R-to-Ф ratios were different in the two organisms. However, linear regression between R and Ф for L. lingvura and U. rigida was stronger (r 2 = 0.814 and 0.313) than the linear regression between R and dry biomass (r 2 = 0.643 and 0.213). The application of Passing–Bablok regression analysis evidenced the high correlation between the results, and the Bland–Altman analysis examined the average difference (“bias”) and limits of agreement between the methods.
The authors are grateful to I. Martínez for her help with the protein determinations. We acknowledge the comments from the reviewers which have contributed greatly to the improvement of this article. This work was accomplished thanks to the Spanish Ministry of Economy and Competitiveness and the Spanish Ministry of Education, Culture and Sports. The research was completed while the senior author was a Ph.D. student in the Doctoral Programme in Oceanography and Global Change at the University of Las Palmas de Gran Canaria.
Compliance with ethical standards
This work was funded by the BIOMBA Project (CTM2012-32729/MAR) Granted to M. Gómez by the Spanish Ministry of Economy and Competitiveness. D. R. Bondyale-Juez received financial support from the FPU Grants from the Spanish Ministry of Education, Culture and Sports. T. T. Packard was supported by TIAA-CREF (USA), Social Security (USA), and by the Canary Islands CEI: Tricontinental Atlantic Campus.
Conflict of interest
The authors have no conflict of interest.
This article does not contain any studies with human participants. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
- Afonso J, Sansón M (1999) Algas, hongos y fanerógamas marinas de las Islas Canarias. Clave Analítica. Materiales didácticos universitarios. Serie Biología 2. Secretario de Publicaciones Universidad de La LagunaGoogle Scholar
- Agardh CA (1823) Species algarum. Lund, SwedenGoogle Scholar
- Cabello-Pasini A, Macías-Carranza V, Abdala R, Korbee N, Figueroa FL (2011) Effect of nitrate concentration and UVR on photosynthesis, respiration, nitrate reductase activity, and phenolic compounds in Ulva rigida (Chlorophyta). J Appl Phycol 23(3):363–369. https://doi.org/10.1007/s10811-010-9548-0 CrossRefGoogle Scholar
- Cleland WW (1967) Enzyme kinetics. Ann Rev Biochem 36(1):77–112. https://doi.org/10.1146/annurev.bi.36.070167.000453 CrossRefGoogle Scholar
- Curl H, Sandberg J (1961) The measurement of dehydrogenase activity in marine organisms. J Mar Res 19(3):123–138Google Scholar
- Dewitte K, Fierens C, Stöckl D, Thienpont LM (2002) Application of the Bland–Altman plot for interpretation of method-comparison studies: a critical investigation of its practice. Clin Chem 48(5):799–801Google Scholar
- Duarte CM, Regaudie-de-Gioux A, Arrieta JM, Delgado-Huertas A, Agustí S (2013) The oligotrophic ocean is heterotrophic. Annu Rev Mar Sci 5:551–569. https://doi.org/10.1146/annurev-marine-121211-172337 CrossRefGoogle Scholar
- Ducklow HW, Doney SC (2013) What is the metabolic state of the oligotrophic ocean? A debate. Annu Rev Mar Sci 5:525–533. https://doi.org/10.1146/annurev-marine-121211-172331 CrossRefGoogle Scholar
- Gómez M (2000) Manual de prácticas de zoología Marina. Departamento de Biología, Universidad de Las Palmas de Gran Canaria, Servicio de Publicaciones y Producción Documental, Gran CanariaGoogle Scholar
- Herrera-Ulibarri A (2013) Identification, abundance and rearing of mysids of Gran Canaria: aplication to laboratory and oceanographic respiratory metabolism studies. Ph.D. dissertation, Universidad de Las Palmas de Gran Canaria, Canary Island, SpainGoogle Scholar
- Linnet K (1999) Limitations of the paired t-test for evaluation of method comparison data. Clin Chem 45(2):314–315Google Scholar
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275Google Scholar
- Lubbers DW, Opitz N (1985) US Patent No. RE31, 879. Washington, DC: US Patent and Trademark OfficeGoogle Scholar
- Mazej Z, Gaberscik A (1999) ETS-activity as a measure of vitality of different macrophyte species. Plant Physiol 39(3):181–186Google Scholar
- McDonald JH (2009) Handbook of biological statistics, vol 2. Sparky House Publishing, Baltimore, pp 191–197Google Scholar
- Nachlas MM, Margulies SI, Seligman AM (1960) A colorimetric method for the estimation of succinic dehydrogenase activity. J Biol Chem 235(2):499–503Google Scholar
- Nelson D, Cox MM (2005) Lehninger principles of biochemistry. WH Freeman and Company, New YorkGoogle Scholar
- Omori M, Ikeda T (1984) Methods in marine zooplankton ecology. xiii, 332 pp. John Wiley. J Mar Biol Assoc UK 65(2):562–562. https://doi.org/10.1017/S0025315400050669
- Opitz N (1986) O2 Optodes for analyzing micro blood samples using thin sensor layers with small O2 capacities and special reflection properties for optical decoupling of sensor and sample. In: Longmuir IS (ed) Oxygen transport to tissue VIII. Advances in experimental medicine and biology, vol 200. Springer, Boston, MAGoogle Scholar
- Packard TT (1985a) Measurement of electron transport activity of marine microplankton. In: Williams PJB, Jannasch HW (eds) Advances in aquatic microbiology. Academic Press, New York, pp 207–261Google Scholar
- Packard TT, Williams PJL (1981) Rates of respiratory oxygen-consumption and electron-transport in surface seawater from the northwest Atlantic. Oceanol Acta 4(3):351–358Google Scholar
- Peltier G, Cournac L (2002) Chlororespiration. Annu Rev Plant Biol 53(1):523–550. https://doi.org/10.1146/annurev.arplant.53.100301.135242 CrossRefGoogle Scholar
- Rutter WJ (1967) Protein determination in embryos. In: Wilt FH, Wessells NK (eds) Methods in developmental biology. Thomas Y. Crowell Co., New York, pp 671–683Google Scholar
- Sars GO (1866) II. Beretning om en i Sommeren 1865 foretagen zoologisk Reise ved Kysterne af Christianias og Christiansands Stifter. Nyt Magazin for Naturvidenskaberne. 15:84–128Google Scholar
- Satpati GG, Pal R (2011) Biochemical composition and lipid characterization of marine green alga Ulva rigida—a nutritional approach. J Algal Biomass Utln 2(4):10–13Google Scholar
- Smith JJ, McFeters GA (1997) Mechanisms of INT (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl tetrazolium chloride), and CTC (5-cyano-2, 3-ditolyl tetrazolium chloride) reduction in Escherichia coli K-12. J Microbiol Methods 29(3):161–175. https://doi.org/10.1016/S0167-7012(97)00036-5 CrossRefGoogle Scholar
- Williams PJ, Del Giorgio PA (2005) Respiration in aquatic ecosystems: history and background. In: Del Giorgio PA, Williams PJ (eds) Respiration in aquatic ecosystems. Oxford University Press, Oxford, pp 1–17Google Scholar
- Williams PJLB, Quay PD, Westberry TK, Behrenfeld MJ (2013) The oligotrophic ocean is autotrophic. Annu Rev Mar Sci 5:535–549. https://doi.org/10.1146/annurev-marine-121211-172335 CrossRefGoogle Scholar