Zebrafish have become a widely used vertebrate model in physiology and reliable measures of their metabolic rate are needed. We have developed a 3D-printed respirometer and swim tunnel system and used it for obtaining accurate measurement of standard metabolic rate (SMR) and maximal, aerobic metabolic rate (MMR) in zebrafish under rest and maximal exercise, respectively. We compared a slow (stepwise) protocol to a fast (continuous) protocol for determining MMR. The fast protocol yielded slightly (but not significantly) higher oxygen consumption rates than the slow protocol and the data, in contrast to the slow protocol, followed a normal distribution. These findings point to the fast protocol as a fast and reliable method for obtaining accurate values of MMR in zebrafish. We make the 3D drawings for printing the system available to researchers, to help streamline the field of metabolic research in zebrafish and other smaller fish species.

Levels of dissolved oxygen in open ocean and coastal waters are decreasing (ocean deoxygenation), with poorly understood effects on marine megafauna. All of the more than 1000 species of elasmobranchs (sharks, skates, and rays) are obligate water breathers, with a variety of life-history strategies and oxygen requirements. This review demonstrates that although many elasmobranchs typically avoid hypoxic water, they also appear capable of withstanding mild to moderate hypoxia with changes in activity, ventilatory responses, alterations to circulatory and hematological parameters, and morphological alterations to gill structures. However, such strategies may be insufficient to withstand severe, progressive, or prolonged hypoxia or anoxia where anaerobic metabolic pathways may be used for limited periods. As water temperatures increase with climate warming, ectothermic elasmobranchs will exhibit elevated metabolic rates and are likely to be less able to tolerate the effects of even mild hypoxia associated with deoxygenation. As a result, sustained hypoxic conditions in warmer coastal or surface-pelagic waters are likely to lead to shifts in elasmobranch distributions. Mass mortalities of elasmobranchs linked directly to deoxygenation have only rarely been observed but are likely underreported. One key concern is how reductions in habitat volume as a result of expanding hypoxia resulting from deoxygenation will influence interactions between elasmobranchs and industrial fisheries. Catch per unit of effort of threatened pelagic sharks by longline fisheries, for instance, has been shown to be higher above oxygen minimum zones compared to adjacent, normoxic regions, and attributed to vertical habitat compression of sharks overlapping with increased fishing effort. How a compound stressor such as marine heatwaves alters vulnerability to deoxygenation remains an open question. With over a third of elasmobranch species listed as endangered, a priority for conservation and management now lies in understanding and mitigating ocean deoxygenation effects in addition to population declines already occurring from overfishing.

The maximum rate at which animals take up oxygen from their environment (ṀO2,max) is a crucial aspect of their physiology and ecology. In fishes, ṀO2,max is commonly quantified by measuring oxygen uptake either during incremental swimming tests or during recovery from an exhaustive chase. In this Commentary, we compile recent studies that apply both techniques to the same fish and show that the two methods typically yield different mean estimates of ṀO2,max for a group of individuals. Furthermore, within a group of fish, estimates of ṀO2,max determined during swimming are poorly correlated with estimates determined during recovery from chasing (i.e. an individual\'s ṀO2,max is not repeatable across methods). One explanation for the lack of agreement is that these methods measure different physiological states, each with their own behavioural, anatomical and biochemical determinants. We propose that these methods are not directly interchangeable but, rather, each is suited to address different questions in fish biology. We suggest that researchers select the method that reflects the biological contexts of their study, and we advocate for the use of accurate terminology that acknowledges the technique used to elevate ṀO2 (e.g. peak ṀO2,swim or peak ṀO2,recovery). If the study\'s objective is to estimate the \'true\' ṀO2,max of an individual or species, we recommend that pilot studies compare methods, preferably using repeated-measures designs. We hope that these recommendations contribute new insights into the causes and consequences of variation in ṀO2,max within and among fish species.

Microorganisms quickly colonise microplastics entering the ocean, forming a biofilm that, if ingested, is consumed with the microplastics. Past research often neglects to expose fish to biofouled microplastics, opting only for clean microplastics despite the low likelihood that fish will encounter clean microplastics. Here, we investigate the physiological impacts of biofouled polyethylene microplastic (300-335 μm) exposure in juvenile fish. Intermittent flow respirometry, antioxidant enzyme activity, and lipid peroxidation were investigated after fish were exposed to clean, biofouled, or no microplastic beads. Fish exposed to biofouled microplastics had a wider aerobic scope than those exposed to clean microplastics while antioxidant enzyme and lipid peroxidation levels were higher in clean microplastics. Clean microplastic exposure indicated higher fitness costs, potentially due to a nutritional advantage of the biofilm or varying bioavailability. These findings highlight the importance of replicating natural factors in exposure experiments when predicting the impacts of increasing pollutants in marine systems.

Bioenergetics is informative for a range of fundamental and applied resource management questions, but findings are often constrained by a lack of ecological realism due to the challenges of remotely estimating key parameters such as metabolic rate. To enable field applications, we conducted a calibration study with smallmouth bass (Micropterus dolomieu, 0.7-2 kg) surgically implanted with accelerometer transmitters and exposed to a ramp-Ucrit swimming protocol in a swim tunnel respirometer across a range of water temperatures (6, 12, 18, and 24°C). There was an exponential increase in fish acceleration with swimming speed, and acceleration per speed was higher in smaller fish and female fish, and at colder temperatures. Mass-specific fish metabolic rate (MO2; mg O2 kg-1 h-1) increased with swimming speed, acceleration, and temperature, and decreased with fish mass, which when combined were strong predictors of MO2. Maximum metabolic rate (MMR) was estimated to peak at 22°C, but maximum sustained swimming speed (Ucrit) remained high at c. 90-100 m s-1 above 20°C, based on second-order polynomial functions. Aerobic scope (AS) estimates peaked at 20°C (>90% AS at 17-24°C; >50% AS at 11-28°C). Males exhibited marginally higher MMR, AS, and Ucrit than females at higher temperatures. Larger fish generally exhibited higher Ucrit, but smaller fish had a marginally broader performance range (AS, Ucrit) among temperatures, benefiting from higher MMR despite a steeper increase in resting metabolic rate with temperature. These findings enable field studies to estimate metabolic metrics of smallmouth bass in situ to characterize their ecological energetics and inform bioenergetics models.

Sinking or floating is the natural state of planktonic organisms and particles in the ocean. Simulating these conditions is critical when making measurements, such as respirometry, because they allow the natural exchange of substrates and products between sinking particles and water flowing around them and prevent organisms that are accustomed to motion from changing their metabolism. We developed a rotating incubator, the RotoBOD (named after its capability to rotate and determine biological oxygen demand, BOD), that uniquely enables automated oxygen measurements in small volumes while keeping the samples in their natural state of suspension. This allows highly sensitive rate measurements of oxygen utilization and subsequent characterization of single particles or small planktonic organisms, such as copepods, jellyfish, or protists. As this approach is nondestructive, it can be combined with several further measurements during and after the incubation, such as stable isotope additions and molecular analyses. This makes the instrument useful for ecologists, biogeochemists, and potentially other user groups such as aquaculture facilities. Here, we present the technical background of our newly developed apparatus and provide examples of how it can be utilized to determine oxygen production and consumption in small organisms and particles.

This study investigated the effectiveness of incorporating sulphur (S) with nitrogen (N) and phosphorus (P) for enhancing microbial activity in diesel-contaminated soil during ex-situ bioremediation. While N and P amendments are commonly used to stimulate indigenous microorganisms, the potential benefits of adding S have received less attention. The study found that historically contaminated soil with a moderate concentration of total petroleum hydrocarbons (TPH; 1270 mg/kg) did not have nutrient limitation, and incubation temperature was found to be more critical for enhancing microbial activity. However, soil spiked with an additional 5000 mg/kg of diesel showed increased activity following NP and NPS amendment. Interestingly, NPS amendment at 10 °C resulted in higher microbial activity than at 20 °C, indicating the potential for a tailored nutrient amendment approach to optimize bioremediation in cold conditions. Overall, this study suggests that incorporating S with N and P can enhance microbial activity in diesel-contaminated soil during ex-situ bioremediation. Furthermore, the study highlights the importance of considering incubation temperature in designing a nutrient amendment approach for bioremediation, especially in cold conditions. These findings can guide the design and implementation of future effective bioremediation strategies for petroleum hydrocarbon-contaminated soil.

Animals routinely encounter environmental (e.g., high temperatures and hypoxia) as well as physiological perturbations (e.g., exercise and digestion) that may threaten homeostasis. However, comparing the relative threat or \"disruptiveness\" imposed by different stressors is difficult, as stressors vary in their mechanisms, effects, and timescales. We exploited the fact that several acute stressors can induce the loss of equilibrium (LOE) in fish to (i) compare the metabolic recovery profiles of three environmentally relevant stressors and (ii) test the concept that LOE could be used as a physiological calibration for the intensity of different stressors. We focused on Etheostoma caeruleum, a species that routinely copes with environmental fluctuations in temperature and oxygen and that relies on burst swimming to relocate and avoid predators, as our model. Using stop-flow (intermittent) respirometry, we tracked the oxygen consumption rate (MO2) as E. caeruleum recovered from LOE induced by hypoxia (PO2 at LOE), warming (critical thermal maximum, CTmax), or exhaustive exercise. Regardless of the stressor used, E. caeruleum recovered rapidly, returning to routine MO2 within ~3 h. Fish recovering from hypoxia and warming had similar maximum MO2, aerobic scopes, recovery time, and total excess post-hypoxia or post-warming oxygen consumption. Though exhaustive exercise induced a greater maximum MO2 and corresponding higher aerobic scope than warming or hypoxia, its recovery profile was otherwise similar to the other stressors, suggesting that \"calibration\" to a physiological state such as LOE may be a viable conceptual approach for investigators interested in questions related to multiple stressors, cross tolerance, and how animals cope with challenges to homeostasis.

Organ function declines with age, and large-scale transcriptomic analyses have highlighted differential aging trajectories across tissues. The mechanisms underlying shared and organ-selective functional changes across the lifespan, however, still remains poorly understood. Given the central role of mitochondria in powering cellular processes needed to maintain tissue health, we therefore undertook a systematic assessment of respiratory activity across 33 different tissues in young (2.5 months) and old (20 months) mice of both sexes. Our high-resolution mitochondrial respiration atlas reveals: 1) within any group of mice, mitochondrial activity varies widely across tissues, with the highest values consistently seen in heart, brown fat, and kidney; 2) biological sex is a significant but minor contributor to mitochondrial respiration, and its contributions are tissue-specific, with major differences seen in the pancreas, stomach, and white adipose tissue; 3) age is a dominant factor affecting mitochondrial activity, especially across different fat depots and skeletal muscle groups, and most brain regions; 4) age-effects can be sex- and tissue-specific, with some of the largest effects seen in pancreas, heart, adipose tissue, and skeletal muscle; and 5) while aging alters the functional trajectories of mitochondria in a majority of tissues, some are remarkably resilient to age-induced changes. Altogether, our data provide the most comprehensive compendium of mitochondrial respiration and illuminate functional signatures of aging across diverse tissues and organ systems.

Lumpfish (Cyclopterus lumpus) mortalities have been reported during the summer at some North Atlantic salmon cage-sites where they serve as \"cleaner fish.\" To better understand this species\' physiology and whether limitations in their metabolic capacity and thermal tolerance can explain this phenomenon, we compared the aerobic scope (AS) of 6°C-acclimated lumpfish (~50 g and 8.8 cm in length at the beginning of experiments) when all individuals (N = 12) were given a chase to exhaustion, a critical swim speed (Ucrit) test, and a critical thermal maximum (CTMax) test (rate of warming 2°C h-1). The Ucrit and CTMax of the lumpfish were 2.36 ± 0.08 body lengths per second and 20.6 ± 0.3°C. The AS of lumpfish was higher during the Ucrit test (206.4 ± 8.5 mg O2 kg-1 h-1) versus that measured in either the CTMax test or after the chase to exhaustion (141.0 ± 15.0 and 124.7 ± 15.5 mg O2 kg-1 h-1, respectively). Maximum metabolic rate (MMR), AS, and \"realistic\" AS (ASR) measured using the three different protocols were not significantly correlated, indicating that measurements of metabolic capacity using one of these methods cannot be used to estimate values that would be obtained using another method. Additional findings include that (1) the lumpfish\'s metabolic capacity is comparable to that of Atlantic cod, suggesting that they are not as \"sluggish\" as previously suggested in the literature, and (2) their CTMax (20.6°C when acclimated to 6°C), in combination with their recently determined ITMax (20.6°C when acclimated to 10°C), indicates that high sea-cage temperatures are unlikely to be the primary cause of lumpfish mortalities at salmon sea-cages during the summer.