Understanding tetrazolium reduction and the importance of substrates in measuring respiratory electron transport activity

Maldonado, F.; Packard, Theodore T.; Gómez, May

doi:10.1016/j.jembe.2012.08.010

Cited by 49 publications

(28 citation statements)

References 71 publications

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“…The calculation of respiration from ETS activity (Packard 1971, King 99 ) and correlation with micro-and mesozooplankton in vivo respiration rates (Packard et al 1974, Kenner & Ahmed 1975, Owens & King 1975 can be useful in calculating the ocean carbon requirement (Packard & Christensen 2004, Packard & Gómez 2013. The rela-tionship between respiration and ETS for the mi croplanktonic population (< 225 µm), shown by Aristegui & Montero (1995) and by Maldonado et al (2012), is not unlike the calibration factors used as proxies for other planktonic metabolic processes, like productivity (Aristegui & Montero 1995). The carbon requirements per unit of zooplankton biomass displayed variability in diel and seasonal vertical distribution patterns.…”

Section: Discussionmentioning

confidence: 99%

“…The rate-limiting step of the system is the oxidation of the coenzyme Q-cytochrome B complex, and it can be measured by its reaction with the artificial electron acceptor 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) (Packard 1971). Many papers have shown good correlation between ETS activity and in vivo respiration, so that ETS activity can be used as a proxy of micro-and mesozooplankton respiration rates (Packard et al 1974, Kenner & Ahmed 1975, Owens & King 1975, Gómez et al 1996, La Ferla et al 1999, Packard & Christensen 2004, Packard & Gómez 2008, 2013, Martínez et al 2010, Maldonado et al 2012.…”

Section: Introductionmentioning

confidence: 99%

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Mesozooplankton carbon requirement in the southern Adriatic Sea: vertical distribution, diel and seasonal variability, relation to particle flux

Minutoli

Granata²,

Brugnano³

et al. 2014

Mar. Ecol. Prog. Ser.

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Mesozooplankton plays a role in particulate organic carbon (POC) remineralisation from the pelagic sinking POC flux down the water column. The vertical distribution (to 1000 m depth), diel variability (during a 24 h cycle) and seasonal differences of the mesozooplankton carbon requirement (µg C g −1 d −1 ), estimated by measuring the activity of the electron transport system, were investigated at a fixed station in the southern Adriatic Sea during November 2006 and April 2007. To estimate the quantitative role of zooplankton in the carbon loss that occurs during organic particle sinking, the zooplankton carbon requirement was compared with organic carbon vertical fluxes measured using a sediment trap. Zooplankton abundance, biomass and community composition were investigated along with the diel vertical distribution of euphausiid migrating groups and the relative importance of crustaceans and gelatinous taxa to the potential community carbon requirement. Day/night zooplankton carbon requirement differences in the upper 100 m were shown in both sampling periods. They were not at all correlated to the vertical distribution pattern of euphausiids but were attributable to the amount of non-living specimens and/or the ratio between gelatinous and crustacean taxa. The mean metabolic requirement in the sampled layers of the water column was lower in November than in April, ranging from 0.02 to 9.10% and from 0.07 to 31.86%, respectively, of the measured carbon losses. In the epi-and mesopelagic zones, the total percent contributions of mesozooplankton respiration to organic carbon losses were 0.18 and 18.45%, respectively, in November and 1.41 and 87.27%, respectively, in April. The higher remineralisation percentages detected in April compared to November could be linked to seasonal differences that involve higher primary production, zooplankton biomass, vertical amount of sinking POC flux and more intense heterotrophic metabolism, which take place mainly in the meso pelagic layer in each period.KEY WORDS: Mesozooplankton · Euphausiids · Carbon requirement · ETS · Sinking POC flux · Adriatic SeaResale or republication not permitted without written consent of the publisher

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Mesozooplankton carbon requirement in the southern Adriatic Sea: vertical distribution, diel and seasonal variability, relation to particle flux

Minutoli

Granata²,

Brugnano³

et al. 2014

Mar. Ecol. Prog. Ser.

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“…We used the enzymatic approach because direct measurements of physiology such as the water-bottle procedures give low spatial resolution and are additionally complicated by artifacts derived from organism manipulation, overcrowding and even starvation when long incubation times are needed (Bidigare, 1983). Enzymatic assays in turn, require the addition of saturating levels of substrates to ensure the specificity of the reaction (Maldonado et al, 2012;Segel, 1993), resulting in a potential measurement which gives the maximum rate (V max ) instead of the actual one. Nevertheless, studies have demonstrated the utility of enzymes as an accurate index of metabolic rates, specially when they are previously calibrated for the surveyed system (e.g., Bode et al, 2013;Hernández-León et al, 1999;Packard, 1985).…”

Section: Introductionmentioning

confidence: 99%

Distribution of zooplankton biomass and potential metabolic activities across the northern Benguela upwelling system

Fernández‐Urruzola

Osma

Packard

et al. 2014

Journal of Marine Systems

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The distribution of zooplankton biomass and potential metabolic rates, in terms of electron transport system (ETS) and glutamate dehydrogenase (GDH), were analyzed along a cross-shelf transect in waters off Namibia. The highly variable dynamics of upwelling filaments promoted short-term fluctuations in the zooplankton biomass and metabolism. Maximum values were characteristically found over the shelf-break, where zooplankton biomass as dry mass (DM) reached peaks of 64.5 mg m −3 within the upper 200 m in late August. Two weeks later, the zooplankton-DM decreased by more than a third (19 mg DM m −3). Zooplankton potential respiration and NH 4 + excretion averaged 234 μmol O 2 m −3 d −1 and 169 μmol NH 4 + m −3 d −1 in the Namibian shelf, respectively. High protein-specific ETS activities even in the low-chlorophyll waters outside the filament suggested a shift into greater omnivory seaward. In this light, zooplankton elemental and isotopic compositions were used to investigate the pelagic food web interactions. They evidenced spatial changes in the carbon resource for zooplankton as well as changes in the form of nitrogen that fueled the biological production in aging advected waters. Overall, both aspects of zooplankton metabolism impacted the primary productivity at a level less than 10% under all the different oceanographic conditions.

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“…A frequently applied biochemical method in soil and freshwater research is the measurement of respiratory electron transport system activity (ETSA) or respiratory potential (Broberg, 1985;G.-Tóth, Langó, Padisák, & Varga, 1994;Garcia, Hernandez, & Costa, 1997;Mersi & Schinner, 1991;Simčič & Mori, 2007;Simčič, Mori, Hossli, Robinson, & Doering, 2015;Trevors, 1984). It measures potential respiration by detecting the activity in the dehydrogenases and cytochromes within respiratory electron transport systems that biochemically control oxygen consumption (Maldonado, Packard, & Gómez, 2012). Because the electron transport system is present in both aerobic and anaerobic microbes and the tetrazolium salt used in the assay is an artificial electron acceptor (replacing O 2 or other naturally occurring acceptors), the technique estimates the total (aerobic and anaerobic) respiratory potential of the microbial community (Maldonado et al, 2012;Zimmerman, 1975).…”

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confidence: 99%

“…It measures potential respiration by detecting the activity in the dehydrogenases and cytochromes within respiratory electron transport systems that biochemically control oxygen consumption (Maldonado, Packard, & Gómez, 2012). Because the electron transport system is present in both aerobic and anaerobic microbes and the tetrazolium salt used in the assay is an artificial electron acceptor (replacing O 2 or other naturally occurring acceptors), the technique estimates the total (aerobic and anaerobic) respiratory potential of the microbial community (Maldonado et al, 2012;Zimmerman, 1975). Previous studies have shown that ETSA correlates highly with oxygen consumption, CO 2 production, and the amount of organic matter in soils or sediments (Cook & Garland, 1997;Garcia et al, 1997;Muri & Simčič, 2004;Simčič & Mori, 2007;Trevors, 1984) and thus is a good estimate of ecosystem functioning.…”

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confidence: 99%

Spatiotemporal heterogeneity of actual and potential respiration in two contrasting floodplains

Mori

Simčić

Brancelj

et al. 2017

Hydrological Processes

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Floodplains are vital components of river ecosystems and play an important role in carbon cycling and storage at catchment and global scales. For efficient river management and conservation, it is critical to understand the functional role of spatiotemporally complex and dynamic habitat mosaics of river floodplains. Unfortunately, the fundamental understanding of mineralization and carbon flux patterns across complex floodplains is still fragmentary. In this study, respiratory potential (i.e., electron transport system activity [ETSA]) was quantified seasonally across different aquatic and terrestrial habitats (wetted channels, gravel bars, islands, riparian forests, and grasslands) of 2 Alpine floodplains differing in climate, altitude, discharge, flow alteration intensity, and land use (Soča [natural flow regime, 12% grassland area] and Urbach [mean annual discharge reduction by 30% due to water abstraction, 69% grassland area]). In situ respiration (R) was measured, and ETSA–R ratios were calculated to examine differences in exploitation intensity of the overall respiratory capacity among floodplain habitats and seasons. ETSA and R provided potential and actual estimates, respectively, of organic matter mineralization in the different floodplain habitats. Hierarchical linear regression across habitat types showed that organic matter, grain sizes <0.063 mm, and water content were the most important predictors of ETSA in both floodplains, and grain sizes 2–0.063 and >8 mm were also highly important for the Soča floodplain. The combination of ETSA and R measurements conducted in contrasting floodplains increased our understanding of the relationships between floodplain habitat heterogeneity, organic matter mineralization and human impacts, that is, structural–functional linkages in floodplains. These data are integral towards predicting changes in floodplain function in response to environmental alterations from increasing human pressures and environmental change.

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Understanding tetrazolium reduction and the importance of substrates in measuring respiratory electron transport activity

Cited by 49 publications

References 71 publications

Mesozooplankton carbon requirement in the southern Adriatic Sea: vertical distribution, diel and seasonal variability, relation to particle flux

Mesozooplankton carbon requirement in the southern Adriatic Sea: vertical distribution, diel and seasonal variability, relation to particle flux

Distribution of zooplankton biomass and potential metabolic activities across the northern Benguela upwelling system

Spatiotemporal heterogeneity of actual and potential respiration in two contrasting floodplains

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