This study compares the critical oxygen saturation (O2crit) levels of the shiner perch Cymatogast... more This study compares the critical oxygen saturation (O2crit) levels of the shiner perch Cymatogaster aggregata obtained using two different methods wherein hypoxia is induced either by the fish's respiration (closed respirometry) or by degassing oxygen with nitrogen (intermittent‐flow respirometry). Fish exhibited loss of equilibrium at a higher O2 saturation in the closed respirometry method when compared with the intermittent‐flow method. Utilization of closed respirometry yielded O2crit measurements that were almost twice as high as those obtained with intermittent‐flow respirometry. The lower hypoxia tolerance in closed respirometry is consistent with additional stress, caused by a build‐up of ammonia and carbon dioxide and a faster rate in dissolved oxygen decline. The results indicate that these two methods of determining hypoxia tolerance in aquatic organisms are not comparable, and that much care should be given to method choice.
Many animals live in groups because of the potential benefits associated with defense and foragin... more Many animals live in groups because of the potential benefits associated with defense and foraging. Group living may also induce a 'calming effect' on individuals, reducing overall metabolic demand. This effect could occur by minimising the need for individual vigilance and reducing stress through social buffering. However, this effect has proved difficult to quantify. We examined the effect of shoaling on metabolism and body condition in the gregarious damselfish Chromis viridis Using a novel respirometry methodology for social species, we found that the presence of shoal-mate visual and olfactory cues led to a reduction in the minimum metabolic rate of individuals. Fish held in isolation for 1 week also exhibited a reduction in body condition when compared with those held in shoals. These results indicate that social isolation as a result of environmental disturbance could have physiological consequences for gregarious species.
Data for Killen et al The Effect of Metabolic Phenotype on Sociability and Social Group Size Preference in a Coral Reef Fish
Although individuals within social groups experience reduced predation risk and find food patches... more Although individuals within social groups experience reduced predation risk and find food patches more consistently, there can be competition for food among group mates. Individuals with a higher standard metabolic rate (SMR) may be less social, to prioritise food acquisition over defense, while greater maximum metabolic rate (MMR) may modulate sociability through increased competitive ability. Therefore, in theory, individuals with a higher SMR may prefer smaller groups and those with greater MMR may prefer larger groups. We examined links among metabolic phenotype, sociability, and choice of group size in the redbelly yellowtail fusilier Caesio cuning. Individuals were exposed to three association tests: (1) a choice between two fish or zero fish; (2) a choice between five fish or zero fish; and (3) a choice between two fish and five fish. The first two tests quantified sociability while the third measured group size choice. Although there was no link between SMR and sociability, fish with a higher MMR were more social than those individuals with a lower MMR. While no correlation was found between MMR and group size choice, there was weak evidence that, if anything, individuals with a higher SMR preferred larger groups, contrary to our hypothesis. As C. cuning is an active fish that spends a large proportion of time operating above SMR, this result could suggest that the links between sociability and SMR may shift depending on the species´ routine behaviour. Links between sociability and MMR may arise if competitive ability allows individuals to obtain resources within groups. Although metabolic traits had no significant influence on group size choice, variation in food availability or predation risk could alter the effects of metabolism on group size choice.
Thermal acclimation of tropical coral reef fishes to global heat waves
eLife
As climate-driven heat waves become more frequent and intense, there is increasing urgency to und... more As climate-driven heat waves become more frequent and intense, there is increasing urgency to understand how thermally sensitive species are responding. Acute heating events lasting days to months may elicit acclimation responses to improve performance and survival. However, the coordination of acclimation responses remains largely unknown for most stenothermal species. We documented the chronology of 18 metabolic and cardiorespiratory changes that occur in the gills, blood, spleen, and muscles when tropical coral reef fishes are thermally stressed (+3.0°C above ambient). Using representative coral reef fishes (Caesio cuning and Cheilodipterus quinquelineatus) separated by >100 million years of evolution and with stark differences in major life-history characteristics (i.e. lifespan, habitat use, mobility, etc.), we show that exposure duration illicited coordinated responses in 13 tissue and organ systems over 5 weeks. The onset and duration of biomarker responses differed betwee...
30 Years of Research on Crown-of-thorns Starfish (1986-2016): Scientific Advances and Emerging Opportunities
Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last fe... more Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last few decades, mostly in accordance with the occurrence of population outbreaks at key locations, such as Australia’s Great Barrier Reef. This review considers advances in our understanding of the biology and ecology of CoTS based on the latest resurgence of research interest, which culminated in this current special issue on the Biology, Ecology and Management of Crown-of-Thorns Starfish. More specifically, this review considers progress against 41 specific research questions posed in the seminal review by P. Moran 30 years ago, as well as exploring new directions for CoTS research. Despite the plethora of research on CoTS (>1,200 research articles), there are persistent knowledge gaps that constrain effective management of outbreaks. Although directly addressing some of these questions will be extremely difficult, there have been considerable advances in understanding the biology ...
1. Metabolic costs associated with parasites should not be limited to established infections. Eve... more 1. Metabolic costs associated with parasites should not be limited to established infections. Even during initial exposure to questing and attacking parasites, hosts can enact behavioural and physiological responses that could also incur metabolic costs. However, few studies have measured these costs directly. Hence, little is known about metabolic costs arising from parasite exposure. 2. Furthermore, no one has yet measured whether and how previous infection history modulates metabolic responses to parasite exposure. 3. Here, using the California killifish Fundulus parvipinnis and its brain-infecting parasite Euhaplorchis californiensis, we quantified how killifish metabolism, behaviour and osmoregulatory phenotype changed upon acute exposure to parasite infectious stages (i.e. cercariae), and with long-term infection. 4. Exposure to cercariae caused both naïve and long-term infected killifish to acutely increase their metabolic rate and activity, indicating detection and response to parasite infectious stages. Additionally, these metabolic and behavioural effects were moderately stronger in long-term infected hosts than naïve killifish, suggesting that hosts may develop learned behavioural responses, nociceptor sensitiza-tion and/or acute immune mechanisms to limit new infections. 5. Although established infection altered the metabolic response to parasite exposure , established infection did not alter standard metabolic rate, routine metabolic rate, maximum metabolic rate, aerobic scope or citrate synthase enzyme activity. 6. Unexpectedly, established infection reduced lactate dehydrogenase enzyme activity in killifish brains and relative Na + /K +-ATPase abundance in gills, suggesting novel mechanisms by which E. californiensis may alter its hosts' behaviour and osmoregulation. 7. Thus, we provide empirical evidence that parasites can disrupt the metabolism of their host both during parasite exposure and after infection is established. This response may be modulated by previous infection history, with probable knock-on effects for host performance, brain energy metabolism, osmoregulation and ecology. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Research conducted on model organisms may be biased due to undetected pathogen infections. Recent... more Research conducted on model organisms may be biased due to undetected pathogen infections. Recently, screening studies discovered high prevalence of the microsporidium Pseudoloma neurophilia in zebrafish (Danio rerio) facilities. This spore-forming unicellular parasite aggregates in brain regions associated with motor function and anxiety, and despite its high occurrence little is known about how sub-clinical infection affects behaviour. Here, we assessed how P. neurophilia infection alters the zebrafish´s response to four commonly used neurobehavioral tests, namely: mirror biting, open field, light/dark preference and social preference, used to quantify aggression, exploration, anxiety, and sociability. Although sociability and aggression remained unaltered, infected hosts exhibited reduced activity, elevated rates of freezing behaviour, and sex-specific effects on exploration. These results indicate that caution is warranted in the interpretation of zebrafish behaviour, particularly since in most cases infection status is unknown. This highlights the importance of comprehensive monitoring procedures to detect sub-clinical infections in laboratory animals. Model animal species (e.g., rodents, invertebrates and fish) are widely used in biomedicine, where study outcomes hinge on reproducibility of the results. Regular health monitoring of these animals has improved over time, as parasites and pathogens (e.g. microparasites, macroparasites, bacteria, viruses) are known to influence animal physiology, immune mechanisms, functional morphology, behaviour, and welfare 1,2. However, monitoring procedures may fail to detect subclinical infections (i.e., exhibiting no external signs of disease), in animals that appear otherwise healthy 3. Thus, undetected infections can inadvertently bias results obtained from these studies, which has repercussions on many research areas, such as biomedicine. The scale of this issue is only just being uncovered. In rodents, for example, Pritchett-Corning et al. 4 reported the prevalence of sixteen commonly undetected pathogens in mice and rats from pharmaceutical, biotechnology, academic, and governmental institutions in North America and Europe. However, the practical impacts of these elusive infectious agents on frequently used experimental assays remain largely unknown. Undetected parasites and pathogens can alter experimental results in model organisms in several ways. Many species of parasites seem to be particularly adapted to affect host neuroendocrine signalling and behaviour in ways which enhance parasite fitness 5-7 , but other aspects of host phenotype are indeed also affected by infection. For example, the intracellular parasite Wolbachia, which is commonly found in laboratory Drosophila spp. colonies , can reduce host egg viability, confound host optimal trait expression (i.e., intra-locus sexual conflict) and alter host circadian rhythms 8-10 , all commonly measured traits in biomedical studies. Similar effects have been observed in rodent model systems. A common infectious agent in rodent facilities is the pathogen murine noro-virus 4,11 , which can induce tissue inflammation and activate cytokine signalling in murine macrophages 12-14. In well-established model animal systems, like Drosophila spp. and rodents (e.g. Mus musculus, Rattus norvegicus), substantial efforts in recent years have focused on how common parasites and pathogens spread within and among laboratory facilities, as well as best practices to remove these infectious agents once established. This work has helped to successfully eliminate and prevent many infections from research facilities, improving both animal welfare and the reproducibility of study outcomes 15,16. However, in newer model organism species (e.g., zebrafish,
Some parasite species alter the behavior of intermediate hosts to promote transmission to the nex... more Some parasite species alter the behavior of intermediate hosts to promote transmission to the next host in the parasite's life cycle. This is the case for Euhaplorchis californiensis, a brain-encysting trematode parasite that causes behavioral changes in the California killifish (Fundulus parvipinnis). These manipulations increase predation by the parasite's final host, piscivorous marsh birds. The mechanisms by which E. californiensis achieves this manipulation remain poorly understood. As E. californiensis cysts reside on the surface of the killifish's brain, discerning regional differences in parasite distribution could indicate mechanisms for host control. In this study, we developed a method for repeated experimental infections. In addition, we measured brain-region specific density using a novel methodology to locate and quantify parasite infection. We show that E. californiensis cysts are non-randomly distributed on the fish brain, aggregating on the diencephalon/mesencephalon region (a brain area involved in controlling reproduction and stress coping) and the rhombencephalon (an area involved in controlling locomotion and basal physiology). Determining causal mechanisms behind this pattern of localization will guide future research examining the neurological mechanisms of parasite-induced host manipulation. These findings suggest that parasites are likely targeting the reproductive, monoaminergic, and locomotor systems to achieve host behavioral manipulation.
Body size and substrate type modulate movement by the western Pacific crown-ofthorns starfish, Acanthaster solaris
PLoS One, 2017
The movement capacity of the crown-of-thorns starfishes (Acanthaster spp.) is a primary determina... more The movement capacity of the crown-of-thorns starfishes (Acanthaster spp.) is a primary determinant of both their distribution and impact on coral assemblages. We quantified individual movement rates for the Pacific crown-of-thorns starfish (Acanthaster solaris) ranging in size from 75±480 mm total diameter, across three different substrates (sand, flat consolidated pavement, and coral rubble) on the northern Great Barrier Reef. The mean (±SE) rate of movement for smaller (<150 mm total diameter) A. solaris was 23.99 ± 1.02 cm/ min and 33.41 ± 1.49 cm/ min for individuals >350 mm total diameter. Mean (±SE) rates of movement varied with substrate type, being much higher on sand (36.53 ± 1.31 cm/ min) compared to consolidated pavement (28.04 ± 1.15 cm/ min) and slowest across coral rubble (17.25 ± 0.63 cm/ min). If average rates of movement measured here can be sustained, in combination with strong directionality, displacement distances of adult A. solaris could range from 250±520 m/ day, depending on the prevailing substrate. Sustained movement of A. solaris is, however, likely to be highly constrained by habitat heterogeneity, energetic constraints, resource availability, and diurnal patterns of activity, thereby limiting their capacity to move between reefs or habitats.
Animals are exposed to variable and rapidly changing environmental flow conditions, such as wind ... more Animals are exposed to variable and rapidly changing environmental flow conditions, such as wind in terrestrial habitats and currents in aquatic systems. For fishes, previous work suggests that individuals exhibit flow-induced changes in aerobic swimming performance. Yet, no one has examined whether similar plasticity is found in fast-start escape responses, which are modulated by anaerobic swimming performance, sensory stimuli and neural control. In this study, we used fish from wild schools of the tropical damselfish Chromis viridis from shallow reefs surrounding Lizard Island in the Great Barrier Reef, Australia. The flow regime at each site was measured to ascertain differences in mean water flow speed and its temporal variability. Swimming and escape behaviour in fish schools were video-recorded in a laminar-flow swim tunnel. Though each school’s swimming behaviour (i.e. alignment and cohesion) was not associated with local flow conditions, traits linked with fast-start performance (particularly turning rate and the distance travelled with the response) were significantly greater in individuals from high-flow habitats. This stronger performance may occur due to a number of mechanisms, such as an in situ training effect or greater selection pressure for faster performance phenotypes in areas with high flow speed.
Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last fe... more Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last few decades, mostly in response to population outbreaks at specific locations. This review considers advances in our understanding of the biology and ecology of CoTS based on the resurgence of research interest, which culminated in this current special issue on the Biology, Ecology and Management of Crown-of-Thorns Starfish. More specifically, this review considers progress in addressing 41 specific research questions posed in a seminal review by P. Moran 30 years ago, as well as exploring new directions for CoTS research. Despite the plethora of research on CoTS (>1200 research articles), there are persistent knowledge gaps that constrain effective management of outbreaks. Although directly addressing some of these questions will be extremely difficult, there have been considerable advances in understanding the biology of CoTS, if not the proximate and ultimate cause(s) of outbreaks. Moving forward, researchers need to embrace new technologies and opportunities to advance our understanding of CoTS biology and behavior, focusing on key questions that will improve effectiveness of management in reducing the frequency and likelihood of outbreaks, if not preventing them altogether
Philosophical Transactions of the Royal Society B, 2017
Individuals of gregarious species often group with conspecifics to which they are phenotypically ... more Individuals of gregarious species often group with conspecifics to which they are phenotypically similar. This among-group assortment has been studied for body size, sex and relatedness. However, the role of physiological traits has been largely overlooked. Here, we discuss mechanisms by which physiological traits—particularly those related to metabolism and locomotor performance—may result in phenotypic assortment not only among but also within animal groups. At the among-group level, varying combinations of passive assortment, active assortment, phenotypic plasticity and selective mortality may generate phenotypic differences among groups. Even within groups, however, individual variation in energy requirements, aerobic and anaerobic capacity, neurological lateralization and tolerance to environmental stressors are likely to produce differences in the spatial location of individuals or associations between group mates with specific physiological phenotypes. Owing to the greater availability of empirical research, we focus on groups of fishes (i.e. shoals and schools). Increased knowledge of physiological mechanisms influencing among- and within-group assortment will enhance our understanding of fundamental concepts regarding optimal group size, predator avoidance, group cohesion, information transfer, life-history strategies and the evolutionary effects of group membership. In a broader perspective, predicting animal responses to environmental change will be impossible without a comprehensive understanding of the physiological basis of the formation and functioning of animal social groups.
In the UK, the Norway lobster (Nephrops norvegicus) supports its most important shellfish fishery... more In the UK, the Norway lobster (Nephrops norvegicus) supports its most important shellfish fishery. Nephrops are sold either whole, or as “tails-only” for the scampi trade. In the “tailing” process, the “head” (cephalothorax) is discarded as waste. A smaller crustacean species, the Antarctic krill Euphasia superba, represents an economically valuable industry, as its extractable oil is sold as a human dietary supplement. The aim of this study was to determine the amount and composition of the oil contained in discarded Nephrops heads and to compare its composition to the oil extracted from krill. Differences due to Geographical variation and seasonal patterns in the amount and composition of lipid were also noted. Results indicated that Nephrops head waste samples collected from more southern locations in Scotland (Clyde Sea area) contained higher levels of oil when compared to samples collected from northern locations in Iceland. Moreover, seasonal differences within the Clyde Sea area in Scotland were also observed, with oil extracted from Nephrops head waste peaking at around 11.5% during the summer months when larger and more mature females were caught by trawl. At this time of the year, the valuable fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) accounted for around 23% of the total fatty acid content in oil extracted from Nephrops head waste. A seasonal effect on EPA content was found, with higher levels obtained in the summer, while no trend was found in DHA percentages. Finally, oil from Nephrops head waste contained a higher proportion of EPA and DHA than krill oil but these fatty acids were more abundantly linked to the neutral lipids rather to than polar lipids. The characterization of lipid that could be extracted from Nephrops head waste should be seen as a first step for the commercial use of a valuable resource currently wasted. This approach is extremely relevant given the current limited supply of EPA and DHA and changes in the Common Fisheries Policy
Atmospheric CO2 is expected to more than double by the end of the century. The resulting changes ... more Atmospheric CO2 is expected to more than double by the end of the century. The resulting changes in ocean chemistry will affect the behaviour, sensory systems and physiology of a range of fish species. Although a number of past studies have examined effects of CO2 in gregarious fishes, most have assessed individuals in social isolation, which can alter individual behaviour and metabolism in social species. Within social groups, a learned familiarity can develop following a prolonged period of interaction between individuals, with fishes preferentially associating with familiar conspecifics because of benefits such as improved social learning and greater foraging opportunities. However, social recognition occurs through detection of shoal-mate cues; hence, it may be disrupted by near-future CO2 conditions. In the present study, we examined the influence of elevated CO2 on shoal familiarity and the metabolic benefits of group living in the gregarious damselfish species the blue-green puller (Chromis viridis). Shoals were acclimated to one of three nominal CO2 treatments: control (450 μatm), mid-CO2 (750 μatm) or high-CO2 (1000 μatm). After a 4–7 day acclimation period, familiarity was examined using a choice test, in which individuals were given the choice to associate with familiar shoal-mates or unfamiliar conspecifics. In control conditions, individuals preferentially associated with familiar shoal-mates. However, this association was lost in both elevated-CO2 treatments. Elevated CO2 did not impact the calming effect of shoaling on metabolism, as measured using an intermittent-flow respirometry methodology for social species following a 17–20 day acclimation period to CO2 treatment. In all CO2 treatments, individuals exhibited a significantly lower metabolic rate when measured in a shoal vs. alone, highlighting the complexity of shoal dynamics and the processes that influence the benefits of shoaling.
Many animals live in groups because of the potential benefits associated with defense and foragin... more Many animals live in groups because of the potential benefits associated with defense and foraging. Group living may also induce a 'calming effect' on individuals, reducing overall metabolic demand. This effect could occur by minimising the need for individual vigilance and reducing stress through social buffering. However, this effect has proved difficult to quantify. We examined the effect of shoaling on metabolism and body condition in the gregarious damselfish Chromis viridis. Using a novel respirometry methodology for social species, we found that the presence of shoal-mate visual and olfactory cues led to a reduction in the minimum metabolic rate of individuals. Fish held in isolation for 1 week also exhibited a reduction in body condition when compared with those held in shoals. These results indicate that social isolation as a result of environmental disturbance could have physiological consequences for gregarious species.
This study compares the critical oxygen saturation (O2crit) levels of the shiner perch Cymatogast... more This study compares the critical oxygen saturation (O2crit) levels of the shiner perch Cymatogaster aggregata obtained using two different methods wherein hypoxia is induced either by the fish's respiration (closed respirometry) or by degassing oxygen with nitrogen (intermittent‐flow respirometry). Fish exhibited loss of equilibrium at a higher O2 saturation in the closed respirometry method when compared with the intermittent‐flow method. Utilization of closed respirometry yielded O2crit measurements that were almost twice as high as those obtained with intermittent‐flow respirometry. The lower hypoxia tolerance in closed respirometry is consistent with additional stress, caused by a build‐up of ammonia and carbon dioxide and a faster rate in dissolved oxygen decline. The results indicate that these two methods of determining hypoxia tolerance in aquatic organisms are not comparable, and that much care should be given to method choice.
Many animals live in groups because of the potential benefits associated with defense and foragin... more Many animals live in groups because of the potential benefits associated with defense and foraging. Group living may also induce a 'calming effect' on individuals, reducing overall metabolic demand. This effect could occur by minimising the need for individual vigilance and reducing stress through social buffering. However, this effect has proved difficult to quantify. We examined the effect of shoaling on metabolism and body condition in the gregarious damselfish Chromis viridis Using a novel respirometry methodology for social species, we found that the presence of shoal-mate visual and olfactory cues led to a reduction in the minimum metabolic rate of individuals. Fish held in isolation for 1 week also exhibited a reduction in body condition when compared with those held in shoals. These results indicate that social isolation as a result of environmental disturbance could have physiological consequences for gregarious species.
Data for Killen et al The Effect of Metabolic Phenotype on Sociability and Social Group Size Preference in a Coral Reef Fish
Although individuals within social groups experience reduced predation risk and find food patches... more Although individuals within social groups experience reduced predation risk and find food patches more consistently, there can be competition for food among group mates. Individuals with a higher standard metabolic rate (SMR) may be less social, to prioritise food acquisition over defense, while greater maximum metabolic rate (MMR) may modulate sociability through increased competitive ability. Therefore, in theory, individuals with a higher SMR may prefer smaller groups and those with greater MMR may prefer larger groups. We examined links among metabolic phenotype, sociability, and choice of group size in the redbelly yellowtail fusilier Caesio cuning. Individuals were exposed to three association tests: (1) a choice between two fish or zero fish; (2) a choice between five fish or zero fish; and (3) a choice between two fish and five fish. The first two tests quantified sociability while the third measured group size choice. Although there was no link between SMR and sociability, fish with a higher MMR were more social than those individuals with a lower MMR. While no correlation was found between MMR and group size choice, there was weak evidence that, if anything, individuals with a higher SMR preferred larger groups, contrary to our hypothesis. As C. cuning is an active fish that spends a large proportion of time operating above SMR, this result could suggest that the links between sociability and SMR may shift depending on the species´ routine behaviour. Links between sociability and MMR may arise if competitive ability allows individuals to obtain resources within groups. Although metabolic traits had no significant influence on group size choice, variation in food availability or predation risk could alter the effects of metabolism on group size choice.
Thermal acclimation of tropical coral reef fishes to global heat waves
eLife
As climate-driven heat waves become more frequent and intense, there is increasing urgency to und... more As climate-driven heat waves become more frequent and intense, there is increasing urgency to understand how thermally sensitive species are responding. Acute heating events lasting days to months may elicit acclimation responses to improve performance and survival. However, the coordination of acclimation responses remains largely unknown for most stenothermal species. We documented the chronology of 18 metabolic and cardiorespiratory changes that occur in the gills, blood, spleen, and muscles when tropical coral reef fishes are thermally stressed (+3.0°C above ambient). Using representative coral reef fishes (Caesio cuning and Cheilodipterus quinquelineatus) separated by >100 million years of evolution and with stark differences in major life-history characteristics (i.e. lifespan, habitat use, mobility, etc.), we show that exposure duration illicited coordinated responses in 13 tissue and organ systems over 5 weeks. The onset and duration of biomarker responses differed betwee...
30 Years of Research on Crown-of-thorns Starfish (1986-2016): Scientific Advances and Emerging Opportunities
Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last fe... more Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last few decades, mostly in accordance with the occurrence of population outbreaks at key locations, such as Australia’s Great Barrier Reef. This review considers advances in our understanding of the biology and ecology of CoTS based on the latest resurgence of research interest, which culminated in this current special issue on the Biology, Ecology and Management of Crown-of-Thorns Starfish. More specifically, this review considers progress against 41 specific research questions posed in the seminal review by P. Moran 30 years ago, as well as exploring new directions for CoTS research. Despite the plethora of research on CoTS (>1,200 research articles), there are persistent knowledge gaps that constrain effective management of outbreaks. Although directly addressing some of these questions will be extremely difficult, there have been considerable advances in understanding the biology ...
1. Metabolic costs associated with parasites should not be limited to established infections. Eve... more 1. Metabolic costs associated with parasites should not be limited to established infections. Even during initial exposure to questing and attacking parasites, hosts can enact behavioural and physiological responses that could also incur metabolic costs. However, few studies have measured these costs directly. Hence, little is known about metabolic costs arising from parasite exposure. 2. Furthermore, no one has yet measured whether and how previous infection history modulates metabolic responses to parasite exposure. 3. Here, using the California killifish Fundulus parvipinnis and its brain-infecting parasite Euhaplorchis californiensis, we quantified how killifish metabolism, behaviour and osmoregulatory phenotype changed upon acute exposure to parasite infectious stages (i.e. cercariae), and with long-term infection. 4. Exposure to cercariae caused both naïve and long-term infected killifish to acutely increase their metabolic rate and activity, indicating detection and response to parasite infectious stages. Additionally, these metabolic and behavioural effects were moderately stronger in long-term infected hosts than naïve killifish, suggesting that hosts may develop learned behavioural responses, nociceptor sensitiza-tion and/or acute immune mechanisms to limit new infections. 5. Although established infection altered the metabolic response to parasite exposure , established infection did not alter standard metabolic rate, routine metabolic rate, maximum metabolic rate, aerobic scope or citrate synthase enzyme activity. 6. Unexpectedly, established infection reduced lactate dehydrogenase enzyme activity in killifish brains and relative Na + /K +-ATPase abundance in gills, suggesting novel mechanisms by which E. californiensis may alter its hosts' behaviour and osmoregulation. 7. Thus, we provide empirical evidence that parasites can disrupt the metabolism of their host both during parasite exposure and after infection is established. This response may be modulated by previous infection history, with probable knock-on effects for host performance, brain energy metabolism, osmoregulation and ecology. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Research conducted on model organisms may be biased due to undetected pathogen infections. Recent... more Research conducted on model organisms may be biased due to undetected pathogen infections. Recently, screening studies discovered high prevalence of the microsporidium Pseudoloma neurophilia in zebrafish (Danio rerio) facilities. This spore-forming unicellular parasite aggregates in brain regions associated with motor function and anxiety, and despite its high occurrence little is known about how sub-clinical infection affects behaviour. Here, we assessed how P. neurophilia infection alters the zebrafish´s response to four commonly used neurobehavioral tests, namely: mirror biting, open field, light/dark preference and social preference, used to quantify aggression, exploration, anxiety, and sociability. Although sociability and aggression remained unaltered, infected hosts exhibited reduced activity, elevated rates of freezing behaviour, and sex-specific effects on exploration. These results indicate that caution is warranted in the interpretation of zebrafish behaviour, particularly since in most cases infection status is unknown. This highlights the importance of comprehensive monitoring procedures to detect sub-clinical infections in laboratory animals. Model animal species (e.g., rodents, invertebrates and fish) are widely used in biomedicine, where study outcomes hinge on reproducibility of the results. Regular health monitoring of these animals has improved over time, as parasites and pathogens (e.g. microparasites, macroparasites, bacteria, viruses) are known to influence animal physiology, immune mechanisms, functional morphology, behaviour, and welfare 1,2. However, monitoring procedures may fail to detect subclinical infections (i.e., exhibiting no external signs of disease), in animals that appear otherwise healthy 3. Thus, undetected infections can inadvertently bias results obtained from these studies, which has repercussions on many research areas, such as biomedicine. The scale of this issue is only just being uncovered. In rodents, for example, Pritchett-Corning et al. 4 reported the prevalence of sixteen commonly undetected pathogens in mice and rats from pharmaceutical, biotechnology, academic, and governmental institutions in North America and Europe. However, the practical impacts of these elusive infectious agents on frequently used experimental assays remain largely unknown. Undetected parasites and pathogens can alter experimental results in model organisms in several ways. Many species of parasites seem to be particularly adapted to affect host neuroendocrine signalling and behaviour in ways which enhance parasite fitness 5-7 , but other aspects of host phenotype are indeed also affected by infection. For example, the intracellular parasite Wolbachia, which is commonly found in laboratory Drosophila spp. colonies , can reduce host egg viability, confound host optimal trait expression (i.e., intra-locus sexual conflict) and alter host circadian rhythms 8-10 , all commonly measured traits in biomedical studies. Similar effects have been observed in rodent model systems. A common infectious agent in rodent facilities is the pathogen murine noro-virus 4,11 , which can induce tissue inflammation and activate cytokine signalling in murine macrophages 12-14. In well-established model animal systems, like Drosophila spp. and rodents (e.g. Mus musculus, Rattus norvegicus), substantial efforts in recent years have focused on how common parasites and pathogens spread within and among laboratory facilities, as well as best practices to remove these infectious agents once established. This work has helped to successfully eliminate and prevent many infections from research facilities, improving both animal welfare and the reproducibility of study outcomes 15,16. However, in newer model organism species (e.g., zebrafish,
Some parasite species alter the behavior of intermediate hosts to promote transmission to the nex... more Some parasite species alter the behavior of intermediate hosts to promote transmission to the next host in the parasite's life cycle. This is the case for Euhaplorchis californiensis, a brain-encysting trematode parasite that causes behavioral changes in the California killifish (Fundulus parvipinnis). These manipulations increase predation by the parasite's final host, piscivorous marsh birds. The mechanisms by which E. californiensis achieves this manipulation remain poorly understood. As E. californiensis cysts reside on the surface of the killifish's brain, discerning regional differences in parasite distribution could indicate mechanisms for host control. In this study, we developed a method for repeated experimental infections. In addition, we measured brain-region specific density using a novel methodology to locate and quantify parasite infection. We show that E. californiensis cysts are non-randomly distributed on the fish brain, aggregating on the diencephalon/mesencephalon region (a brain area involved in controlling reproduction and stress coping) and the rhombencephalon (an area involved in controlling locomotion and basal physiology). Determining causal mechanisms behind this pattern of localization will guide future research examining the neurological mechanisms of parasite-induced host manipulation. These findings suggest that parasites are likely targeting the reproductive, monoaminergic, and locomotor systems to achieve host behavioral manipulation.
Body size and substrate type modulate movement by the western Pacific crown-ofthorns starfish, Acanthaster solaris
PLoS One, 2017
The movement capacity of the crown-of-thorns starfishes (Acanthaster spp.) is a primary determina... more The movement capacity of the crown-of-thorns starfishes (Acanthaster spp.) is a primary determinant of both their distribution and impact on coral assemblages. We quantified individual movement rates for the Pacific crown-of-thorns starfish (Acanthaster solaris) ranging in size from 75±480 mm total diameter, across three different substrates (sand, flat consolidated pavement, and coral rubble) on the northern Great Barrier Reef. The mean (±SE) rate of movement for smaller (<150 mm total diameter) A. solaris was 23.99 ± 1.02 cm/ min and 33.41 ± 1.49 cm/ min for individuals >350 mm total diameter. Mean (±SE) rates of movement varied with substrate type, being much higher on sand (36.53 ± 1.31 cm/ min) compared to consolidated pavement (28.04 ± 1.15 cm/ min) and slowest across coral rubble (17.25 ± 0.63 cm/ min). If average rates of movement measured here can be sustained, in combination with strong directionality, displacement distances of adult A. solaris could range from 250±520 m/ day, depending on the prevailing substrate. Sustained movement of A. solaris is, however, likely to be highly constrained by habitat heterogeneity, energetic constraints, resource availability, and diurnal patterns of activity, thereby limiting their capacity to move between reefs or habitats.
Animals are exposed to variable and rapidly changing environmental flow conditions, such as wind ... more Animals are exposed to variable and rapidly changing environmental flow conditions, such as wind in terrestrial habitats and currents in aquatic systems. For fishes, previous work suggests that individuals exhibit flow-induced changes in aerobic swimming performance. Yet, no one has examined whether similar plasticity is found in fast-start escape responses, which are modulated by anaerobic swimming performance, sensory stimuli and neural control. In this study, we used fish from wild schools of the tropical damselfish Chromis viridis from shallow reefs surrounding Lizard Island in the Great Barrier Reef, Australia. The flow regime at each site was measured to ascertain differences in mean water flow speed and its temporal variability. Swimming and escape behaviour in fish schools were video-recorded in a laminar-flow swim tunnel. Though each school’s swimming behaviour (i.e. alignment and cohesion) was not associated with local flow conditions, traits linked with fast-start performance (particularly turning rate and the distance travelled with the response) were significantly greater in individuals from high-flow habitats. This stronger performance may occur due to a number of mechanisms, such as an in situ training effect or greater selection pressure for faster performance phenotypes in areas with high flow speed.
Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last fe... more Research on the coral-eating crown-of-thorns starfish (CoTS) has waxed and waned over the last few decades, mostly in response to population outbreaks at specific locations. This review considers advances in our understanding of the biology and ecology of CoTS based on the resurgence of research interest, which culminated in this current special issue on the Biology, Ecology and Management of Crown-of-Thorns Starfish. More specifically, this review considers progress in addressing 41 specific research questions posed in a seminal review by P. Moran 30 years ago, as well as exploring new directions for CoTS research. Despite the plethora of research on CoTS (>1200 research articles), there are persistent knowledge gaps that constrain effective management of outbreaks. Although directly addressing some of these questions will be extremely difficult, there have been considerable advances in understanding the biology of CoTS, if not the proximate and ultimate cause(s) of outbreaks. Moving forward, researchers need to embrace new technologies and opportunities to advance our understanding of CoTS biology and behavior, focusing on key questions that will improve effectiveness of management in reducing the frequency and likelihood of outbreaks, if not preventing them altogether
Philosophical Transactions of the Royal Society B, 2017
Individuals of gregarious species often group with conspecifics to which they are phenotypically ... more Individuals of gregarious species often group with conspecifics to which they are phenotypically similar. This among-group assortment has been studied for body size, sex and relatedness. However, the role of physiological traits has been largely overlooked. Here, we discuss mechanisms by which physiological traits—particularly those related to metabolism and locomotor performance—may result in phenotypic assortment not only among but also within animal groups. At the among-group level, varying combinations of passive assortment, active assortment, phenotypic plasticity and selective mortality may generate phenotypic differences among groups. Even within groups, however, individual variation in energy requirements, aerobic and anaerobic capacity, neurological lateralization and tolerance to environmental stressors are likely to produce differences in the spatial location of individuals or associations between group mates with specific physiological phenotypes. Owing to the greater availability of empirical research, we focus on groups of fishes (i.e. shoals and schools). Increased knowledge of physiological mechanisms influencing among- and within-group assortment will enhance our understanding of fundamental concepts regarding optimal group size, predator avoidance, group cohesion, information transfer, life-history strategies and the evolutionary effects of group membership. In a broader perspective, predicting animal responses to environmental change will be impossible without a comprehensive understanding of the physiological basis of the formation and functioning of animal social groups.
In the UK, the Norway lobster (Nephrops norvegicus) supports its most important shellfish fishery... more In the UK, the Norway lobster (Nephrops norvegicus) supports its most important shellfish fishery. Nephrops are sold either whole, or as “tails-only” for the scampi trade. In the “tailing” process, the “head” (cephalothorax) is discarded as waste. A smaller crustacean species, the Antarctic krill Euphasia superba, represents an economically valuable industry, as its extractable oil is sold as a human dietary supplement. The aim of this study was to determine the amount and composition of the oil contained in discarded Nephrops heads and to compare its composition to the oil extracted from krill. Differences due to Geographical variation and seasonal patterns in the amount and composition of lipid were also noted. Results indicated that Nephrops head waste samples collected from more southern locations in Scotland (Clyde Sea area) contained higher levels of oil when compared to samples collected from northern locations in Iceland. Moreover, seasonal differences within the Clyde Sea area in Scotland were also observed, with oil extracted from Nephrops head waste peaking at around 11.5% during the summer months when larger and more mature females were caught by trawl. At this time of the year, the valuable fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) accounted for around 23% of the total fatty acid content in oil extracted from Nephrops head waste. A seasonal effect on EPA content was found, with higher levels obtained in the summer, while no trend was found in DHA percentages. Finally, oil from Nephrops head waste contained a higher proportion of EPA and DHA than krill oil but these fatty acids were more abundantly linked to the neutral lipids rather to than polar lipids. The characterization of lipid that could be extracted from Nephrops head waste should be seen as a first step for the commercial use of a valuable resource currently wasted. This approach is extremely relevant given the current limited supply of EPA and DHA and changes in the Common Fisheries Policy
Atmospheric CO2 is expected to more than double by the end of the century. The resulting changes ... more Atmospheric CO2 is expected to more than double by the end of the century. The resulting changes in ocean chemistry will affect the behaviour, sensory systems and physiology of a range of fish species. Although a number of past studies have examined effects of CO2 in gregarious fishes, most have assessed individuals in social isolation, which can alter individual behaviour and metabolism in social species. Within social groups, a learned familiarity can develop following a prolonged period of interaction between individuals, with fishes preferentially associating with familiar conspecifics because of benefits such as improved social learning and greater foraging opportunities. However, social recognition occurs through detection of shoal-mate cues; hence, it may be disrupted by near-future CO2 conditions. In the present study, we examined the influence of elevated CO2 on shoal familiarity and the metabolic benefits of group living in the gregarious damselfish species the blue-green puller (Chromis viridis). Shoals were acclimated to one of three nominal CO2 treatments: control (450 μatm), mid-CO2 (750 μatm) or high-CO2 (1000 μatm). After a 4–7 day acclimation period, familiarity was examined using a choice test, in which individuals were given the choice to associate with familiar shoal-mates or unfamiliar conspecifics. In control conditions, individuals preferentially associated with familiar shoal-mates. However, this association was lost in both elevated-CO2 treatments. Elevated CO2 did not impact the calming effect of shoaling on metabolism, as measured using an intermittent-flow respirometry methodology for social species following a 17–20 day acclimation period to CO2 treatment. In all CO2 treatments, individuals exhibited a significantly lower metabolic rate when measured in a shoal vs. alone, highlighting the complexity of shoal dynamics and the processes that influence the benefits of shoaling.
Many animals live in groups because of the potential benefits associated with defense and foragin... more Many animals live in groups because of the potential benefits associated with defense and foraging. Group living may also induce a 'calming effect' on individuals, reducing overall metabolic demand. This effect could occur by minimising the need for individual vigilance and reducing stress through social buffering. However, this effect has proved difficult to quantify. We examined the effect of shoaling on metabolism and body condition in the gregarious damselfish Chromis viridis. Using a novel respirometry methodology for social species, we found that the presence of shoal-mate visual and olfactory cues led to a reduction in the minimum metabolic rate of individuals. Fish held in isolation for 1 week also exhibited a reduction in body condition when compared with those held in shoals. These results indicate that social isolation as a result of environmental disturbance could have physiological consequences for gregarious species.
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Papers by Lauren Nadler
is, however, likely to be highly constrained by habitat heterogeneity, energetic constraints, resource availability, and diurnal patterns of activity, thereby limiting their capacity to move between reefs or habitats.
from shallow reefs surrounding Lizard Island in the Great Barrier Reef, Australia. The flow regime at each site was measured to ascertain differences in mean water flow speed and its temporal variability. Swimming and escape behaviour in fish schools were video-recorded in a laminar-flow swim tunnel. Though each school’s swimming behaviour (i.e. alignment and cohesion) was not associated with local flow conditions, traits linked with fast-start performance (particularly turning rate and the distance travelled with the response) were significantly greater in individuals from high-flow habitats. This stronger performance may occur due to a number of mechanisms, such as an in situ training effect or greater selection pressure for faster
performance phenotypes in areas with high flow speed.
Management of Crown-of-Thorns Starfish. More specifically, this review considers progress in addressing
41 specific research questions posed in a seminal review by P. Moran 30 years ago, as well as exploring new directions for CoTS research. Despite the plethora of research on CoTS (>1200 research articles), there are persistent knowledge gaps that constrain effective management of outbreaks. Although directly addressing some of these questions will be extremely difficult, there have been considerable advances in understanding the biology of CoTS, if not the proximate and ultimate cause(s) of outbreaks. Moving forward, researchers need to embrace new technologies and opportunities to
advance our understanding of CoTS biology and behavior, focusing on key questions that will improve effectiveness of management in reducing the frequency and likelihood of outbreaks, if not preventing them altogether
traits has been largely overlooked. Here, we discuss mechanisms by which physiological traits—particularly those related to metabolism and locomotor performance—may result in phenotypic assortment not only among but also within animal groups. At the among-group level, varying combinations of passive assortment, active assortment, phenotypic plasticity and selective mortality may generate phenotypic differences among groups.
Even within groups, however, individual variation in energy requirements, aerobic and anaerobic capacity, neurological lateralization and tolerance to environmental stressors are likely to produce differences in the spatial location of individuals or associations between group mates with specific physiological phenotypes. Owing to the greater availability of empirical research, we focus on groups of fishes (i.e. shoals and schools). Increased knowledge of physiological mechanisms influencing among- and within-group assortment will enhance our understanding of fundamental concepts regarding optimal group size, predator avoidance, group cohesion, information
transfer, life-history strategies and the evolutionary effects of group membership. In a broader perspective, predicting animal responses to environmental change will be impossible without a comprehensive understanding
of the physiological basis of the formation and functioning of animal social groups.
Euphasia superba, represents an economically valuable industry, as its extractable oil is sold as a human dietary supplement. The aim of this study was to determine the amount and composition of the oil contained in discarded Nephrops heads and to compare its composition to the oil extracted from krill. Differences due to Geographical variation and seasonal patterns in the amount and composition of lipid were also noted. Results indicated that Nephrops head waste samples collected from more
southern locations in Scotland (Clyde Sea area) contained higher levels of oil when compared to samples collected from northern locations in Iceland. Moreover, seasonal differences within the Clyde Sea area in Scotland were also observed, with oil extracted from Nephrops head waste peaking at
around 11.5% during the summer months when larger and more mature females were caught by trawl. At this time of the year, the valuable fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid
(DHA) accounted for around 23% of the total fatty acid content in oil extracted from Nephrops head waste. A seasonal effect on EPA content was found, with higher levels obtained in the summer, while no trend was found in DHA percentages. Finally, oil from Nephrops head waste contained
a higher proportion of EPA and DHA than krill oil but these fatty acids were more abundantly linked to the neutral lipids rather to than polar lipids. The characterization of lipid that could be extracted from Nephrops head waste should be seen as a first step for the commercial use of a valuable resource currently wasted. This approach is extremely relevant given the current limited supply of EPA and DHA and changes in the Common Fisheries Policy
will affect the behaviour, sensory systems and physiology of a range of fish species. Although a number of past studies
have examined effects of CO2 in gregarious fishes, most have assessed individuals in social isolation, which can alter individual
behaviour and metabolism in social species. Within social groups, a learned familiarity can develop following a prolonged
period of interaction between individuals, with fishes preferentially associating with familiar conspecifics because
of benefits such as improved social learning and greater foraging opportunities. However, social recognition occurs
through detection of shoal-mate cues; hence, it may be disrupted by near-future CO2 conditions. In the present study, we
examined the influence of elevated CO2 on shoal familiarity and the metabolic benefits of group living in the gregarious
damselfish species the blue-green puller (Chromis viridis). Shoals were acclimated to one of three nominal CO2 treatments:
control (450 μatm), mid-CO2 (750 μatm) or high-CO2 (1000 μatm). After a 4–7 day acclimation period, familiarity was examined
using a choice test, in which individuals were given the choice to associate with familiar shoal-mates or unfamiliar
conspecifics. In control conditions, individuals preferentially associated with familiar shoal-mates. However, this association
was lost in both elevated-CO2 treatments. Elevated CO2 did not impact the calming effect of shoaling on metabolism, as
measured using an intermittent-flow respirometry methodology for social species following a 17–20 day acclimation period
to CO2 treatment. In all CO2 treatments, individuals exhibited a significantly lower metabolic rate when measured in a
shoal vs. alone, highlighting the complexity of shoal dynamics and the processes that influence the benefits of shoaling.
is, however, likely to be highly constrained by habitat heterogeneity, energetic constraints, resource availability, and diurnal patterns of activity, thereby limiting their capacity to move between reefs or habitats.
from shallow reefs surrounding Lizard Island in the Great Barrier Reef, Australia. The flow regime at each site was measured to ascertain differences in mean water flow speed and its temporal variability. Swimming and escape behaviour in fish schools were video-recorded in a laminar-flow swim tunnel. Though each school’s swimming behaviour (i.e. alignment and cohesion) was not associated with local flow conditions, traits linked with fast-start performance (particularly turning rate and the distance travelled with the response) were significantly greater in individuals from high-flow habitats. This stronger performance may occur due to a number of mechanisms, such as an in situ training effect or greater selection pressure for faster
performance phenotypes in areas with high flow speed.
Management of Crown-of-Thorns Starfish. More specifically, this review considers progress in addressing
41 specific research questions posed in a seminal review by P. Moran 30 years ago, as well as exploring new directions for CoTS research. Despite the plethora of research on CoTS (>1200 research articles), there are persistent knowledge gaps that constrain effective management of outbreaks. Although directly addressing some of these questions will be extremely difficult, there have been considerable advances in understanding the biology of CoTS, if not the proximate and ultimate cause(s) of outbreaks. Moving forward, researchers need to embrace new technologies and opportunities to
advance our understanding of CoTS biology and behavior, focusing on key questions that will improve effectiveness of management in reducing the frequency and likelihood of outbreaks, if not preventing them altogether
traits has been largely overlooked. Here, we discuss mechanisms by which physiological traits—particularly those related to metabolism and locomotor performance—may result in phenotypic assortment not only among but also within animal groups. At the among-group level, varying combinations of passive assortment, active assortment, phenotypic plasticity and selective mortality may generate phenotypic differences among groups.
Even within groups, however, individual variation in energy requirements, aerobic and anaerobic capacity, neurological lateralization and tolerance to environmental stressors are likely to produce differences in the spatial location of individuals or associations between group mates with specific physiological phenotypes. Owing to the greater availability of empirical research, we focus on groups of fishes (i.e. shoals and schools). Increased knowledge of physiological mechanisms influencing among- and within-group assortment will enhance our understanding of fundamental concepts regarding optimal group size, predator avoidance, group cohesion, information
transfer, life-history strategies and the evolutionary effects of group membership. In a broader perspective, predicting animal responses to environmental change will be impossible without a comprehensive understanding
of the physiological basis of the formation and functioning of animal social groups.
Euphasia superba, represents an economically valuable industry, as its extractable oil is sold as a human dietary supplement. The aim of this study was to determine the amount and composition of the oil contained in discarded Nephrops heads and to compare its composition to the oil extracted from krill. Differences due to Geographical variation and seasonal patterns in the amount and composition of lipid were also noted. Results indicated that Nephrops head waste samples collected from more
southern locations in Scotland (Clyde Sea area) contained higher levels of oil when compared to samples collected from northern locations in Iceland. Moreover, seasonal differences within the Clyde Sea area in Scotland were also observed, with oil extracted from Nephrops head waste peaking at
around 11.5% during the summer months when larger and more mature females were caught by trawl. At this time of the year, the valuable fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid
(DHA) accounted for around 23% of the total fatty acid content in oil extracted from Nephrops head waste. A seasonal effect on EPA content was found, with higher levels obtained in the summer, while no trend was found in DHA percentages. Finally, oil from Nephrops head waste contained
a higher proportion of EPA and DHA than krill oil but these fatty acids were more abundantly linked to the neutral lipids rather to than polar lipids. The characterization of lipid that could be extracted from Nephrops head waste should be seen as a first step for the commercial use of a valuable resource currently wasted. This approach is extremely relevant given the current limited supply of EPA and DHA and changes in the Common Fisheries Policy
will affect the behaviour, sensory systems and physiology of a range of fish species. Although a number of past studies
have examined effects of CO2 in gregarious fishes, most have assessed individuals in social isolation, which can alter individual
behaviour and metabolism in social species. Within social groups, a learned familiarity can develop following a prolonged
period of interaction between individuals, with fishes preferentially associating with familiar conspecifics because
of benefits such as improved social learning and greater foraging opportunities. However, social recognition occurs
through detection of shoal-mate cues; hence, it may be disrupted by near-future CO2 conditions. In the present study, we
examined the influence of elevated CO2 on shoal familiarity and the metabolic benefits of group living in the gregarious
damselfish species the blue-green puller (Chromis viridis). Shoals were acclimated to one of three nominal CO2 treatments:
control (450 μatm), mid-CO2 (750 μatm) or high-CO2 (1000 μatm). After a 4–7 day acclimation period, familiarity was examined
using a choice test, in which individuals were given the choice to associate with familiar shoal-mates or unfamiliar
conspecifics. In control conditions, individuals preferentially associated with familiar shoal-mates. However, this association
was lost in both elevated-CO2 treatments. Elevated CO2 did not impact the calming effect of shoaling on metabolism, as
measured using an intermittent-flow respirometry methodology for social species following a 17–20 day acclimation period
to CO2 treatment. In all CO2 treatments, individuals exhibited a significantly lower metabolic rate when measured in a
shoal vs. alone, highlighting the complexity of shoal dynamics and the processes that influence the benefits of shoaling.