Supplemental oxygen does not improve growth but can enhance reproductive capacity of fish.

Fish tend to grow faster as the climate warms but attain a smaller adult body size following an earlier age at sexual maturation. Despite the apparent ubiquity of this phenomenon, termed the temperature-size rule (TSR), heated scientific debates have revealed a poor understanding of the underlying mechanisms. At the centre of these debates are prominent but marginally tested hypotheses which implicate some form of 'oxygen limitation' as the proximate cause. Here, we test the role of oxygen limitation in the TSR by rearing juvenile Galaxias maculatus for a full year in current-day (15°C) and forecasted (20°C) summer temperatures while providing half of each temperature group with supplemental oxygen (hyperoxia). True to the TSR, fish in the warm treatments grew faster and reached sexual maturation earlier than their cooler conspecifics. Yet, despite supplemental oxygen significantly increasing maximum oxygen uptake rate, our findings contradict leading hypotheses by showing that the average size at sexual maturation and the adult body size did not differ between normoxia and hyperoxia groups. We did, however, discover that hyperoxia extended the reproductive window, independent of fish size and temperature. We conclude that the intense resource investment in reproduction could expose a bottleneck where oxygen becomes a limiting factor.

Evidence for energy reallocation, not oxygen limitation, driving the deceleration in growth of adult fish.

The lifetime growth of almost all fishes follows a biphasic relationship - juvenile growth is rapid and adult growth subsequently decelerates. For a trend that is so ubiquitous, there is no general agreement as to the underlying mechanisms causing adult growth to decelerate. Ongoing theories argue that adult growth slows because either the gills fail to supply the body with surplus oxygen needed for continued somatic gain (i.e. oxygen limited), or sexual maturation induces a switch in energy allocation towards reproduction and away from growth (i.e. energy limited). Here, we empirically tested these notions by tracking the individual growth trajectories of ∼100 female Galaxias maculatus, ranging in size, during their first 3 months of adulthood. At a summer temperature of 20°C, we provided subsets of fish with additional energy (fed once versus twice a day), supplementary oxygen (normoxia versus hyperoxia), or a combination of the two, to assess whether we could change the trajectory of adult growth. We found that growth improved marginally with additional energy, yet remained unaffected by supplementary oxygen, thereby providing evidence for a role for energy reallocation in the deceleration of adult growth. Interestingly, additional dietary energy had a disproportionately larger effect on the growth of fish that matured at a greater size, revealing size-dependent variance in energy acquisition and/or allocation budgets at summer temperatures. Overall, these findings contribute towards understanding the mechanisms driving widespread declines in the body size of fish with climate warming.

Chronic experimental hyperoxia elevates aerobic scope: a valid method to test for physiological oxygen limitations in fish.

Experimental hyperoxia has been shown to enhance the maximum oxygen uptake capacity of fishes under acute conditions, potentially offering an avenue to test prominent physiological hypotheses attempting to explain impacts of climate warming on fish populations (e.g., gill-oxygen limitation driving declines in fish size). Such benefits of experimental hyperoxia must persist under chronic conditions if it is to provide a valid manipulation to test the relevant hypotheses, yet the long-term benefits of experimental hyperoxia to oxygen uptake capacity have not been examined. Here, the authors measured aerobic metabolic performance of Galaxias maculatus upon acute exposure to hyperoxia (150% air saturation) and after 5 months of acclimation, at both 15°C and 20°C. Acute hyperoxia elevated aerobic scope by 74%-94% relative to normoxic controls, and an elevation of 58%-73% persisted after 5 months of hyperoxia acclimation. When hyperoxia-acclimated fish were acutely transitioned back to normoxia, they maintained superior aerobic performance compared with normoxic controls, suggesting an acclimation of the underlying metabolic structures/processes. In demonstrating the long-term benefits of experimental hyperoxia on the aerobic performance of a fish, the authors encourage the use of such approaches to disentangle the role of oxygen in driving the responses of fish populations to climate warming.

Behavioural temperature regulation is a low priority in a coral reef fish (Plectropomus leopardus): insights from a novel behavioural thermoregulation system.

Current understanding of behavioural thermoregulation in aquatic ectotherms largely stems from systems such as 'shuttle boxes', which are generally limited in their capacity to test large-bodied species. Here, we introduce a controlled system that allows large aquatic ectotherms to roam freely in a tank at sub-optimal temperatures, using thermal refuges to increase body temperature to their thermal optimum as desired. Of the 10 coral grouper (Plectropomus leopardus; length ∼400 mm) implanted with thermal loggers, three fish maintained themselves at the ambient tank temperature of 17.5-20.5°C for the entire 2-4 days of the trial. Of the other seven fish, body temperature never exceeded ∼21.5°C, which was well below the temperature available in the thermal refuges (∼31°C) and below the species' optimal temperature of ∼27°C. This study adds to a growing literature documenting an unexpected lack of behavioural thermoregulation in aquatic ectotherms in controlled, heterothermal environments.

Paths towards greater consensus building in experimental biology.

In a recent editorial, the Editors-in-Chief of Journal of Experimental Biology argued that consensus building, data sharing, and better integration across disciplines are needed to address the urgent scientific challenges posed by climate change. We agree and expand on the importance of cross-disciplinary integration and transparency to improve consensus building and advance climate change research in experimental biology. We investigated reproducible research practices in experimental biology through a review of open data and analysis code associated with empirical studies on three debated paradigms and for unrelated studies published in leading journals in comparative physiology and behavioural ecology over the last 10 years. Nineteen per cent of studies on the three paradigms had open data, and 3.2% had open code. Similarly, 12.1% of studies in the journals we examined had open data, and 3.1% had open code. Previous research indicates that only 50% of shared datasets are complete and re-usable, suggesting that fewer than 10% of studies in experimental biology have usable open data. Encouragingly, our results indicate that reproducible research practices are increasing over time, with data sharing rates in some journals reaching 75% in recent years. Rigorous empirical research in experimental biology is key to understanding the mechanisms by which climate change affects organisms, and ultimately promotes evidence-based conservation policy and practice. We argue that a greater adoption of open science practices, with a particular focus on FAIR (Findable, Accessible, Interoperable, Re-usable) data and code, represents a much-needed paradigm shift towards improved transparency, cross-disciplinary integration, and consensus building to maximize the contributions of experimental biologists in addressing the impacts of environmental change on living organisms.

The use of internal heart rate loggers in determining cardiac breakpoints of fish.

As marine environments are influenced by global warming there is a need to thoroughly understand the relationship between physiological limits and temperature in fish. One quick screening method of a physiological thermal tipping point is the temperature at which maximum heart rate (ƒHmax) can no longer scale predictably with warming and is referred to as the Arrhenius break temperature (TAB). The use of this method has been successful for freshwater fish by using external electrodes to detect an electrocardiogram (ECG), however, the properties of this equipment pose challenges in salt water when evaluating marine fish. To overcome these challenges, this study aimed to explore the potential use of implantable heart rate loggers to quantify the TAB of Chrysoblephus laticeps, a marine Sparid, following the ECG method protocols where ƒHmax is monitored over an acute warming event and the TAB is subsequently identified using a piece-wise linear regression model. Of the nine experimental fish, only five (56%) returned accurate ƒHmax data. The TAB of successful trials was identified each time and ranged from 18.09 to 20.10 °C. This study therefore provides evidence that implantable heart rate loggers can estimate TAB of fish which can be applied to many marine species.

Implantation, orientation and validation of a commercially produced heart-rate logger for use in a perciform teleost fish.

Quantifying how the heart rate of ectothermic organisms responds to environmental conditions (e.g. water temperature) is important information to quantify their sensitivity to environmental change. Heart rate studies have typically been conducted in lab environments where fish are confined. However, commercially available implantable heart rate biologgers provide the opportunity to study free-swimming fish. Our study aimed to determine the applicability of an implantable device, typically used on fusiform-shaped fish (e.g. salmonids), for a perciform fish where morphology and anatomy prevent ventral incisions normally used on fusiform-shaped fish. We found that ventrolateral incisions allowed placement near the heart, but efficacy of the loggers was sensitive to their orientation and the positioning of the electrodes. Electrocardiogram detection, signal strength and subsequent heart rate readings were strongly influenced by logger orientation with a significant effect on the quality and quantity of heart rate recordings. We provide details on the surgical procedures and orientation to guide future heart rate biologger studies on perciform-shaped fish.