Rapid physiological and transcriptomic changes associated with oxygen delivery in larval anemonefish suggest a role in adaptation to life on hypoxic coral reefs.

Connectivity of coral reef fish populations relies on successful dispersal of a pelagic larval phase. Pelagic larvae must exhibit high swimming abilities to overcome ocean and reef currents, but once settling onto the reef, larvae transition to endure habitats that become hypoxic at night. Therefore, coral reef fish larvae must rapidly and dramatically shift their physiology over a short period of time. Taking an integrative, physiological approach, using swimming respirometry, and examining hypoxia tolerance and transcriptomics, we show that larvae of cinnamon anemonefish (Amphiprion melanopus) rapidly transition between "physiological extremes" at the end of their larval phase. Daily measurements of swimming larval anemonefish over their entire early development show that they initially have very high mass-specific oxygen uptake rates. However, oxygen uptake rates decrease midway through the larval phase. This occurs in conjunction with a switch in haemoglobin gene expression and increased expression of myoglobin, cytoglobin, and neuroglobin, which may all contribute to the observed increase in hypoxia tolerance. Our findings indicate that critical ontogenetic changes in the gene expression of oxygen-binding proteins may underpin the physiological mechanisms needed for successful larval recruitment to reefs.

How hot is too hot? Thermal tolerance, performance, and preference in juvenile mangrove whiprays, Urogymnus granulatus.

Mangrove habitats can serve as nursery areas for sharks and rays. Such environments can be thermally dynamic and extreme; yet, the physiological and behavioural mechanisms sharks and rays use to exploit such habitats are understudied. This study aimed to define the thermal niche of juvenile mangrove whiprays, Urogymnus granulatus. First, temperature tolerance limits were determined via the critical thermal maximum (CTMax) and minimum (CTMin) of mangrove whiprays at summer acclimation temperatures (28 °C), which were 17.5 °C and 39.9 °C, respectively. Then, maximum and routine oxygen uptake rates (MO2max and MO2routine, respectively), post-exercise oxygen debt, and recovery were estimated at current (28 °C) and heatwave (32 °C) temperatures, revealing moderate temperature sensitivities (i.e., Q10) of 2.4 (MO2max) and 1.6 (MO2routine), but opposing effects on post-exercise oxygen uptake. Finally, body temperatures (Tb) of mangrove whiprays were recorded using external temperature loggers, and environmental temperatures (Te) were recorded using stationary temperature loggers moored in three habitat zones (mangrove, reef flat, and reef crest). As expected, environmental temperatures varied between sites depending on depth. Individual mangrove whiprays presented significantly lower Tb relative to Te during the hottest times of the day. Electivity analysis showed tagged individuals selected temperatures from 24.0 to 37.0 °C in habitats that ranged from 21.1 to 43.5 °C. These data demonstrate that mangrove whiprays employ thermotaxic behaviours and a thermally insensitive aerobic metabolism to thrive in thermally dynamic and extreme habitats. Tropical nursery areas may, therefore, offer important thermal refugia for young rays. However, these tropical nursery areas could become threatened by mangrove and coral habitat loss, and climate change.

Importance of health assessments for conservation in noncaptive wildlife.

Wildlife health assessments help identify populations at risk of starvation, disease, and decline from anthropogenic impacts on natural habitats. We conducted an overview of available health assessment studies in noncaptive vertebrates and devised a framework to strategically integrate health assessments in population monitoring. Using a systematic approach, we performed a thorough assessment of studies examining multiple health parameters of noncaptive vertebrate species from 1982 to 2020 (n = 261 studies). We quantified trends in study design and diagnostic methods across taxa with generalized linear models, bibliometric analyses, and visual representations of study location versus biodiversity hotspots. Only 35% of studies involved international or cross-border collaboration. Countries with both high and threatened biodiversity were greatly underrepresented. Species that were not listed as threatened on the International Union for Conservation of Nature Red List represented 49% of assessed species, a trend likely associated with the regional focus of most studies. We strongly suggest following wildlife health assessment protocols when planning a study and using statistically adequate sample sizes for studies establishing reference ranges. Across all taxa blood analysis (89%), body composition assessments (81%), physical examination (72%), and fecal analyses (24% of studies) were the most common methods. A conceptual framework to improve design and standardize wildlife health assessments includes guidelines on the experimental design, data acquisition and analysis, and species conservation planning and management implications. Integrating a physiological and ecological understanding of species resilience toward threatening processes will enable informed decision making regarding the conservation of threatened species.

Importancia de los exámenes diagnósticos para la conservación de fauna silvestre Resumen Los exámenes diagnósticos de fauna silvestre ayudan a identificar poblaciones en riesgo por desnutrición, enfermedades infecciosas y disminución poblacional, causadas por impactos antropogénicos. Revisamos los estudios disponibles que llevaron a cabo exámenes diagnósticos en fauna silvestre y diseñamos un marco de trabajo para integrar dichos exámenes en monitoreos poblacionales. Empleando un enfoque sistemático, evaluamos aquellos estudios que examinaban múltiples indicadores de salud en vertebrados no cautivos entre 1982 y 2020 (n = 261 estudios). Cuantificamos las tendencias estadísticas, clasificadas por taxones, del diseño del estudio y de los métodos diagnósticos usando modelos lineales generalizados, análisis bibliométricos y representaciones visuales del lugar de estudio versus los hotspots (puntos calientes) de biodiversidad. Sólo el 35% de los estudios incluían colaboraciones internacionales o transfronterizas, y los países ricos en biodiversidad y especies amenazadas estaban gravemente subrepresentados. Las especies no clasificadas como amenazadas en la Lista Roja de la Unión Internacional para la Conservación de la Naturaleza representaban el 49% de las especies examinadas; una tendencia posiblemente asociada al enfoque regional de la mayoría de los estudios. Recomendamos encarecidamente seguir protocolos diagnósticos y manuales de técnicas del estudio de la fauna silvestre, además de usar tamaños muestrales estadísticamente adecuados al establecer rangos de referencia. Los métodos diagnósticos más comunes para todos los taxones fueronanálisis sanguíneos (89%), evaluaciones de composición corporal (81%), exámenes físicos (72%) y análisis fecales (24% de los estudios). Presentamos un marco conceptual para mejorar y estandarizar los exámenes diagnósticos en estudios de fauna silvestre; dicho marco incluye guías para el diseño experimental, para la obtención y el análisis de datos, y para elaborar planes de acción para especies amenazadas. La combinación de conocimientos fisiológicos y ecológicos, relacionados con la resiliencia biológica de especies amenazadas, facilitará una toma de decisiones eficiente para el manejo y para la conservación de la biodiversidad.

A novel hyperbaric swimming respirometer allows the simulation of varying swimming depths in fish respirometry studies.

The understanding of swimming physiology and knowledge on the metabolic costs of swimming are important for assessing effects of environmental factors on migratory behavior. Swim tunnels are the most common experimental setups for measuring swimming performance and oxygen uptake rates in fishes; however, few can realistically simulate depth and the changes in hydrostatic pressure that many fishes experience, e.g. during diel vertical migrations. Here, we present a new hyperbaric swimming respirometer (HSR) that can simulate depths of up to 80 m. The system consists of three separate, identical swimming tunnels, each with a volume of 205 L, a control board and a storage tank with water treatment. The swimming chamber of each tunnel has a length of 1.40 m and a diameter of 20 cm. The HSR uses the principle of intermittent-flow respirometry and has here been tested with female European eels (Anguilla anguilla). Various pressure, temperature and flow velocity profiles can be programmed, and the effect on metabolic activity and oxygen consumption can be assessed. Thus, the HSR provides opportunities to study the physiology of fish during swimming in a simulated depth range that corresponds to many inland, coastal and shelf waters.

Parasite infection directly impacts escape response and stress levels in fish.

Parasites can account for a substantial proportion of the biomass in marine communities. As such, parasites play a significant ecological role in ecosystem functioning via host interactions. Unlike macropredators, such as large piscivores, micropredators, such as parasites, rarely cause direct mortality. Rather, micropredators impose an energetic tax, thus significantly affecting host physiology and behaviour via sublethal effects. Recent research suggests that infection by gnathiid isopods (Crustacea) causes significant physiological stress and increased mortality rates. However, it is unclear whether infection causes changes in the behaviours that underpin escape responses or changes in routine activity levels. Moreover, it is poorly understood whether the cost of gnathiid infection manifests as an increase in cortisol. To investigate this, we examined the effect of experimental gnathiid infection on the swimming and escape performance of a newly settled coral reef fish and whether infection led to increased cortisol levels. We found that micropredation by a single gnathiid caused fast-start escape performance and swimming behaviour to significantly decrease and cortisol levels to double. Fast-start escape performance is an important predictor of recruit survival in the wild. As such, altered fitness-related traits and short-term stress, perhaps especially during early life stages, may result in large scale changes in the number of fish that successfully recruit to adult populations.

Thermal impacts on the growth, development and ontogeny of critical swimming speed in Atlantic herring larvae.

Increases in swimming ability have a profound influence on larval fish growth and survival by increasing foraging success, predator avoidance and the ability to favorably influence transport. Understanding how development and environmental factors combine to influence swimming performance in aquatic organisms is particularly important during the transition from viscous to inertial environments. We measured the growth, development and ontogenetic changes in critical swimming speed (Ucrit) in Atlantic herring (Clupea harengus) larvae reared at three temperatures (7, 11, 15°C). Temperature had a significant effect on growth rates (from 0.21 at 7°C to 0.34mm·d(-1) at 15°C), and larval morphology-at-length (increased dry weight (DW), body height and developmental rate at warmer temperatures). Temperature-dependent differences in morphology influenced swimming performance (e.g. the exponential increase in Ucrit with increasing body size was faster at warmer temperatures). Larvae entered the transition to an inertial environment (Reynolds numbers ≥300) at body lengths between 15 (15°C) and 17mm (7°C). Inter-individual differences in Ucrit were not related to nutritional condition (RNA·DNA(-1) or DNA·DW(-1)), but were negatively correlated to length-at-age, suggesting a trade-off between growth rate and locomotor activity. The Ucrit data from this and previously published studies suggest that Atlantic herring pass through four activity phases: 1) yolk-sac (<0.6cm·s(-1)), 2) pre-flexion (0.6-3.0cm·s(-1), temperature effect changes with body size), 3) post-flexion (up to 6-8cm·s(-1), Q10~1.8-2.0), 4) juvenile-adult period (20-170cm·s(-1)).

Habitat complexity and predator odours impact on the stress response and antipredation behaviour in coral reef fish.

Mass coral bleaching events coupled with local stressors have caused regional-scale loss of corals on reefs globally. Following the loss of corals, the structural complexity of these habitats is often reduced. By providing shelter, obscuring visual information, or physically impeding predators, habitat complexity can influence predation risk and the perception of risk by prey. Yet little is known on how habitat complexity and risk assessment interact to influence predator-prey interactions. To better understand how prey's perception of threats may shift in degraded ecosystems, we reared juvenile Pomacentrus chrysurus in environments of various habitat complexity levels and then exposed them to olfactory risk odours before simulating a predator strike. We found that the fast-start escape responses were enhanced when forewarned with olfactory cues of a predator and in environments of increasing complexity. However, no interaction between complexity and olfactory cues was observed in escape responses. To ascertain if the mechanisms used to modify these escape responses were facilitated through hormonal pathways, we conducted whole-body cortisol analysis. Cortisol concentrations interacted with habitat complexity and risk odours, such that P. chrysurus exhibited elevated cortisol levels when forewarned with predator odours, but only when complexity levels were low. Our study suggests that as complexity is lost, prey may more appropriately assess predation risk, likely as a result of receiving additional visual information. Prey's ability to modify their responses depending on the environmental context suggests that they may be able to partly alleviate the risk of increased predator-prey interactions as structural complexity is reduced.

Limiting motorboat noise on coral reefs boosts fish reproductive success.

Anthropogenic noise impacts are pervasive across taxa, ecosystems and the world. Here, we experimentally test the hypothesis that protecting vulnerable habitats from noise pollution can improve animal reproductive success. Using a season-long field manipulation with an established model system on the Great Barrier Reef, we demonstrate that limiting motorboat activity on reefs leads to the survival of more fish offspring compared to reefs experiencing busy motorboat traffic. A complementary laboratory experiment isolated the importance of noise and, in combination with the field study, showed that the enhanced reproductive success on protected reefs is likely due to improvements in parental care and offspring length. Our results suggest noise mitigation could have benefits that carry through to the population-level by increasing adult reproductive output and offspring growth, thus helping to protect coral reefs from human impacts and presenting a valuable opportunity for enhancing ecosystem resilience.

Automated flow control of a multi-lane swimming chamber for small fishes indicates species-specific sensitivity to experimental protocols.

In fishes, swimming performance is considered an important metric to measure fitness, dispersal and migratory abilities. The swimming performance of individual larval fishes is often integrated into models to make inferences on how environmental parameters affect population-level dynamics (e.g. connectivity). However, little information exists regarding how experimental protocols affect the swimming performance of marine fish larvae. In addition, the technical setups used to measure larval fish swimming performance often lack automation and accurate control of water quality parameters and flow velocity. In this study, we automated the control of multi-lane swimming chambers for small fishes by developing an open-source algorithm. This automation allowed us to execute repeatable flow scenarios and reduce operator interference and inaccuracies in flow velocity typically associated with manual control. Furthermore, we made structural modifications to a prior design to reduce the areas of lower flow velocity. We then validated the flow dynamics of the new chambers using computational fluid dynamics and particle-tracking software. The algorithm provided an accurate alignment between the set and measured flow velocities and we used it to test whether faster critical swimming speed (U crit) protocols (i.e. shorter time intervals and higher velocity increments) would increase U crit of early life stages of two tropical fish species [4-10-mm standard length (SL)]. The U crit of barramundi (Lates calcarifer) and cinnamon anemonefish (Amphiprion melanopus) increased linearly with fish length, but in cinnamon anemonefish, U crit started to decrease upon metamorphosis. Swimming protocols using longer time intervals (more than 2.5 times increase) negatively affected U crit in cinnamon anemonefish but not in barramundi. These species-specific differences in swimming performance highlight the importance of testing suitable U crit protocols prior to experimentation. The automated control of flow velocity will create more accurate and repeatable data on swimming performance of larval fishes. Integrating refined measurements into individual-based models will support future research on the effects of environmental change.

Physiology can contribute to better understanding, management, and conservation of coral reef fishes.

Coral reef fishes, like many other marine organisms, are affected by anthropogenic stressors such as fishing and pollution and, owing to climate change, are experiencing increasing water temperatures and ocean acidification. Against the backdrop of these various stressors, a mechanistic understanding of processes governing individual organismal performance is the first step for identifying drivers of coral reef fish population dynamics. In fact, physiological measurements can help to reveal potential cause-and-effect relationships and enable physiologists to advise conservation management by upscaling results from cellular and individual organismal levels to population levels. Here, we highlight studies that include physiological measurements of coral reef fishes and those that give advice for their conservation. A literature search using combined physiological, conservation and coral reef fish key words resulted in ~1900 studies, of which only 99 matched predefined requirements. We observed that, over the last 20 years, the combination of physiological and conservation aspects in studies on coral reef fishes has received increased attention. Most of the selected studies made their physiological observations at the whole organism level and used their findings to give conservation advice on population dynamics, habitat use or the potential effects of climate change. The precision of the recommendations differed greatly and, not surprisingly, was least concrete when studies examined the effects of projected climate change scenarios. Although more and more physiological studies on coral reef fishes include conservation aspects, there is still a lack of concrete advice for conservation managers, with only very few published examples of physiological findings leading to improved management practices. We conclude with a call to action to foster better knowledge exchange between natural scientists and conservation managers to translate physiological findings more effectively in order to obtain evidence-based and adaptive management strategies for the conservation of coral reef fishes.

Projected habitat loss for Atlantic herring in the Baltic Sea.

Projected, climate-driven changes in rainfall patterns are expected to alter the salinity (S) of estuaries and larger brackish water bodies, such as the Baltic Sea. Some marine fish larvae are potentially more sensitive to low salinity than older stages, hence we compared the low salinity tolerance of Atlantic herring (Clupea harengus) larvae at the individual and population levels including four populations in the North and Baltic Seas. Acute low salinity tolerance was similar (S = 1.9-2.7) across populations and increased with increasing body size. Based on this physiological threshold and a regionally down-scaled climate model, spawning habitats in the northern and eastern Baltic Sea are projected to be largely unsuitable for herring by 2100. Although adaptive mechanisms may attenuate the effect in some species, the limited physiological tolerance of fish larvae will remain an important bottleneck for the persistence of marine fish populations in brackish waters undergoing climate-driven freshening.

Direct Effects of Microalgae and Protists on Herring (Clupea harengus) Yolk Sac Larvae.

This study investigated effects of microalgae (Rhodomonas baltica) and heterotrophic protists (Oxyrrhis marina) on the daily growth, activity, condition and feeding success of Atlantic herring (Clupea harengus) larvae from hatch, through the end of the endogenous (yolk sac) period. Yolk sac larvae were reared in the presence and absence of microplankton and, each day, groups of larvae were provided access to copepods. Larvae reared with microalgae and protists exhibited precocious (2 days earlier) and ≥ 60% increased feeding incidence on copepods compared to larvae reared in only seawater (SW). In the absence and presence of microalgae and protists, life span and growth trajectories of yolk sac larvae were similar and digestive enzyme activity (trypsin) and nutritional condition (RNA-DNA ratio) markedly declined in all larvae directly after yolk sac depletion. Thus, microplankton promoted early feeding but was not sufficient to alter life span and growth during the yolk sac phase. Given the importance of early feeding, field programs should place greater emphasis on the protozooplankton-ichthyoplankton link to better understand match-mismatch dynamics and bottom-up drivers of year class success in marine fish.