Respirometry provides a direct measure of an organism\'s O2 consumption rate (VO2), which is a significant component of its metabolic rate (energy expenditure). Amongst ants, variations in lifespan between different social castes (such as workers and queens) can be substantial, varying depending to the species. As metabolic rate is higher in short-living species, we aimed to determine how VO2 and longevity may have coevolved within ant casts. Measuring VO2 in such tiny animal models can be challenging, and as a first methodological step, we validate the use of a Clark electrode, initially designed for measuring mitochondrial respiration control pathways, for assessing VO2 in ants within a sealed chamber. This was done by comparing it with stop-flow VO2 and CO2 production, using a traditional indirect calorimetry device. The global aim is to provide a reliable protocol to conduct accurate comparisons of metabolic rates within and among ant species. As expected, using Clark electrode entails high time resolution and revealed that queens and workers exhibited discontinuous respiration, with episodes of apnea lasting up to 20 min.

Conventional wastewater treatment plants (WWTPs) are not designed for the abatement of antibiotics, and their effluents are one of the main entry ways of these emerging contaminants to the aquatic environment, causing major concern due to their toxicity, persistence, and bioaccumulation. When wastewater containing antibiotics enters the bioreactor, they can impact microbial communities of the activated sludge, affecting biodegradation processes of organic matter and nutrients. There is scarce information about the effect of activated carbon on the activated sludge within the bioreactor in presence of antibiotics. In light of this, the effect of representative antibiotics, ciprofloxacin (CIP), nalidixic acid (NAL), and erythromycin (ERY), on the performance of a conventional activated sludge of a WWTP was analyzed by respirometry with and without activated carbon. NAL and ERY negatively affected the net heterotrophic biomass growth rate (r\'x,H), with reduction percentages of 26%-90% and 31%-81%, respectively. The addition of activated carbon mitigated this effect, especially for ERY, with increments of even 8% in the r\'x,H for the hybrid process when working with 5 ppm of ERY and 80 ppm of activated carbon compared with the value in the absence of antibiotic and activated carbon. This effect was attributed to the enhanced retention of ERY, in comparison to NAL, on the surface of the activated carbon, probably due to its higher molecular size and affinity towards the activated carbon (log Kow = 3.06). This effect was more marked at low sludge retention times (below 8 days). PRACTITIONER POINTS: Ciprofloxacin (CIP), nalidixic acid (NAL), and erythromycin (ERY) were studied. NAL and ERY exerted negative impact on heterotrophic growth rate. Effect of antibiotics on microorganisms in the presence of activated carbon was studied. Activated carbon was mainly relevant for ERY due to its adsorption retention. Enhancement by activated carbon was more significant at low sludge retention times.

Social interactions can sometimes be a source of stress, but social companions can also ameliorate and buffer against stress. Stress and metabolism are closely linked, but the degree to which social companions modulate metabolic responses during stressful situations-and whether such effects differ depending on social rank-is poorly understood. To investigate this question, we studied Neolamprologus pulcher, a group-living cichlid fish endemic to Lake Tanganyika and measured the metabolic responses of dominant and subordinate individuals when they were either visible or concealed from one another. When individuals could see each other, subordinates had lower maximum metabolic rates and tended to take longer to recover following an exhaustive chase compared with dominants. In contrast, metabolic responses of dominants and subordinates did not differ when individuals could not see one another. These findings suggest that the presence of a dominant individual has negative metabolic consequences for subordinates, even in stable social groups with strong prosocial relationships.

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.