Analysing tropical elasmobranch blood samples in the field: blood stability during storage and validation of the HemoCue (R) haemoglobin analyser

Blood samples collected from wild-caught fishes can provide important information regarding the effects of capture (and thus post-release survival) as well as other stressors. Unfortunately, blood samples often cannot be analysed immediately upon sampling, and blood parameters (e.g. blood oxygen levels and acid-base parameters) are known to change with storage duration due to the metabolic activity of the red blood cells. We obtained blood samples from both untreated and stressed individuals of both blacktip reef shark (Carcharhinus melanopterus) and sicklefin lemon shark (Negaprion acutidens) to determine the effects of storage duration on blood pH, haematocrit and haemoglobin concentration ([Hb]). We found no significant effects after storage on ice for up to 180 minutes. Moreover, to validate the usability of a HemoCue haemoglobin analyser (a point-of-care device), we compared data from this device to [Hb] determined using the cyanomethaemoglobin method with blood samples from 10 individuals from each of the aforementioned species as well as epaulette shark (Hemiscyllium ocellatum). Values from the HemoCue consistently overestimated [Hb], and we therefore developed the necessary correction equations. The correction equations were not statistically different among the three elasmobranch species within the biologically relevant range but did differ from published corrections developed using blood from temperate teleost fishes. Although the HemoCue is useful in field situations, development of species-specific calibration equations may be necessary to ensure the reliability of inter-species comparisons of blood [Hb]. Together, these data should increase confidence in haematological stress indicators in elasmobranch fishes, measurements of which are critical for understanding the impact of anthropogenic stressors on these ecologically important species

Combined Effects of Acute Temperature Change and Elevated pCO2 on the Metabolic Rates and Hypoxia Tolerances of Clearnose Skate (Rostaraja eglanteria), Summer Flounder (Paralichthys dentatus), and Thorny Skate (Amblyraja radiata)

Understanding how rising temperatures, ocean acidification, and hypoxia affect the performance of coastal fishes is essential to predicting species-specific responses to climate change. Although a population’s habitat influences physiological performance, little work has explicitly examined the multi-stressor responses of species from habitats differing in natural variability. Here, clearnose skate (Rostaraja eglanteria) and summer flounder (Paralichthys dentatus) from mid-Atlantic estuaries, and thorny skate (Amblyraja radiata) from the Gulf of Maine, were acutely exposed to current and projected temperatures (20, 24, or 28 °C; 22 or 30 °C; and 9, 13, or 15 °C, respectively) and acidification conditions (pH 7.8 or 7.4). We tested metabolic rates and hypoxia tolerance using intermittent-flow respirometry. All three species exhibited increases in standard metabolic rate under an 8 °C temperature increase (Q10 of 1.71, 1.07, and 2.56, respectively), although this was most pronounced in the thorny skate. At the lowest test temperature and under the low pH treatment, all three species exhibited significant increases in standard metabolic rate (44–105%; p \u3c 0.05) and decreases in hypoxia tolerance (60–84% increases in critical oxygen pressure; p \u3c 0.05). This study demonstrates the interactive effects of increasing temperature and changing ocean carbonate chemistry are species-specific, the implications of which should be considered within the context of habitat. Associated dataset: Gail D. Schweiterman, Daniel P. Crear et al. 2019. Metabolic Rates and Hypoxia Tolerences of clearnose skate (Rostaraja eglanteria), summer flounder (Paralichthys dentatus), and thorny skate (Amblyraja radiata) https://doi.org/10.25773/qmew-c18

The power struggle: assessing interacting global change stressors via experimental studies on sharks

Ocean warming and acidification act concurrently on marine ectotherms with the potential for detrimental, synergistic effects; yet, effects of these stressors remain understudied in large predatory fishes, including sharks. We tested for behavioural and physiological responses of blacktip reef shark (Carcharhinus melanopterus) neonates to climate change relevant changes in temperature (28 and 31 °C) and carbon dioxide partial pressures (pCO2; 650 and 1050 µatm) using a fully factorial design. Behavioural assays (lateralisation, activity level) were conducted upon 7–13 days of acclimation, and physiological assays (hypoxia tolerance, oxygen uptake rates, acid–base and haematological status) were conducted upon 14–17 days of acclimation. Haematocrit was higher in sharks acclimated to 31 °C than to 28 °C. Significant treatment effects were also detected for blood lactate and minimum oxygen uptake rate; although, these observations were not supported by adequate statistical power. Inter-individual variability was considerable for all measured traits, except for haematocrit. Moving forward, studies on similarly ‘hard-to-study’ species may account for large inter-individual variability by increasing replication, testing larger, yet ecologically relevant, differences in temperature and pCO2, and reducing measurement error. Robust experimental studies on elasmobranchs are critical to meaningfully assess the threat of global change stressors in these data-deficient species

Metabolic Rates and Hypoxia Tolerences of clearnose skate (Rostaraja eglanteria), summer flounder (Paralichthys dentatus), and thorny skate (Amblyraja radiata)

These data were collected following methods described in the associated publication: LINK “Combined Effects of Acute Temperature Change and Elevated pCO2 on the Metabolic Rates and Hypoxia Tolerances of Clearnose Skate (Rostaraja eglanteria), Summer Flounder (Paralichthys dentatus), and Thorny Skate (Amblyraja radiata)”. Schweiterman, G.D. et al. 2019 Biology, 8(3), 56

The impacts of warming and hypoxia on the performance of an obligate ram ventilator

Climate change is causing the warming and deoxygenation of coastal habitats like Chesapeake Bay that serve as important nursery habitats for many marine fish species. As conditions continue to change, it is important to understand how these changes impact individual species\u27 behavioral and metabolic performance. The sandbar shark (Carcharhinus plumbeus) is an obligate ram-ventilating apex predator whose juveniles use Chesapeake Bay as a nursery ground up to 10 years of age. The objective of this study was to measure juvenile sandbar shark metabolic and behavioral performance as a proxy for overall performance (i.e. fitness or success) when exposed to warm and hypoxic water. Juvenile sandbar sharks (79.5-113.5 cm total length) were collected from an estuary along the eastern shore of Virginia and returned to lab where they were fitted with an accelerometer, placed in a respirometer and exposed to varying temperatures and oxygen levels. Juvenile sandbar shark overall performance declined substantially at 32 degrees C or when dissolved oxygen concentration was reduced below 3.5 mg l(-1) (51% oxygen saturation between 24-32 degrees C). As the extent of warm hypoxic water increases in Chesapeake Bay, we expect that the available sandbar shark nursery habitat will be reduced, which may negatively impact the population of sandbar sharks in the western Atlantic as well as the overall health of the ecosystem within Chesapeake Bay

Analysing tropical elasmobranch blood samples in the field: blood stability during storage and validation of the HemoCue® haemoglobin analyser

Blood samples collected from wild-caught fishes can provide important information regarding the effects of capture (and thus post-release survival) as well as other stressors. Unfortunately, blood samples often cannot be analysed immediately upon sampling, and blood parameters (e.g. blood oxygen levels and acid–base parameters) are known to change with storage duration due to the metabolic activity of the red blood cells. We obtained blood samples from both untreated and stressed individuals of both blacktip reef shark (Carcharhinus melanopterus) and sicklefin lemon shark (Negaprion acutidens) to determine the effects of storage duration on blood pH, haematocrit and haemoglobin concentration ([Hb]). We found no significant effects after storage on ice for up to 180 minutes. Moreover, to validate the usability of a HemoCue haemoglobin analyser (a point-of-care device), we compared data from this device to [Hb] determined using the cyanomethaemoglobin method with blood samples from 10 individuals from each of the aforementioned species as well as epaulette shark (Hemiscyllium ocellatum). Values from the HemoCue consistently overestimated [Hb], and we therefore developed the necessary correction equations. The correction equations were not statistically different among the three elasmobranch species within the biologically relevant range but did differ from published corrections developed using blood from temperate teleost fishes. Although the HemoCue is useful in field situations, development of species-specific calibration equations may be necessary to ensure the reliability of inter-species comparisons of blood [Hb]. Together, these data should increase confidence in haematological stress indicators in elasmobranch fishes, measurements of which are critical for understanding the impact of anthropogenic stressors on these ecologically important species

Thermally insensitive physiological performance allows neonatal sharks to use coastal habitats as nursery areas

Coastal sharks can use shallow, nearshore habitats as nursery areas, which is a behaviour that may increase fitness. The ecological benefits of shark nursery areas are well studied; yet the physiological mechanisms that enable sharks to exploit coastal habitats, especially those that experience extreme and dynamic temperatures, remain understudied. We hypothesised that neonatal sharks are able to use thermally dynamic coastal habitats as nursery areas because temperature does not strongly affect their physiology. To test this hypothesis, we defined patterns of nursery area use and temperature-dependent physiological performance in 2 reef shark species. First, we determined whether 10 sites around the island of Moorea, French Polynesia, satisfied nursery area criteria for neonate populations of blacktip reef sharks Carcharhinus melanopterus and sicklefin lemon sharks Negaprion acutidens using 5 consecutive years of abundance surveys. We then quantified effects of thermal exposure in situ on growth in recaptured individuals and quantified the temperature dependence of metabolic rate ex situ using respirometry. We found several potential C. melanopterus nursery areas, but during different sampling years, and identified 1 N. acutidens nursery area that remained consistent during the entire 5 yr study. In support of our hypothesis, growth and metabolic performance were not strongly affected by temperature in either species. Thus, thermally insensitive physiological performance may be a trait that elasmobranchs exhibit in thermally variable coastal habitats, including shark nursery areas. Together, this approach demonstrates how physiological and ecological concepts complement each other to improve our understanding of nursery area use in coastal shark populations

Swimming depth of dolphinfish (Coryphaena hippurus) associated and unassociated with fish aggregating devices

Dolphinfish (Coryphaena hippurus), large pelagic predators and important fishery targets, frequently associate with floating debris or manmade fish aggregating devices (FADs). We tagged 8 dolphinfish with pressure-sensitive ultrasonic transmitters and actively tracked individuals continuously for up to 40 h to elucidate the vertical movement patterns and differences between FADassociated (FAD-A) and FAD-unassociated (FAD-U) fish. Four additional fish were equipped with acoustic transmitters and passively monitored for several days with receivers attached to FADs. When not associated with FADs, dolphinfish used the upper 75-100 m of the water column during the day and made descents up to 160 m during the night. In contrast, FAD-A fish generally stayed within the upper 10 m of the water column and tended to make deeper excursions during the day rather than at night. Water temperature data from expendable bathythermo-graphs deployed during active tracking showed that fish only descended to depths where temperatures were <= 3 degrees C cooler than the uniform-temperature surface layer. The use of vertical behavior to determine whether a dolphinfish is associated or not with a floating object opens the possibility for new, large-scale research aimed at investigating the role of floating objects in the ecosystem inhabited by this species and at assessing the impacts of FADs on its ecology

Seawater carbonate chemistry and behaviour, physiological metrics in blacktip reef shark (Carcharhinus melanopterus)

Ocean warming and acidification act concurrently on marine ectotherms with the potential for detrimental, synergistic effects; yet, effects of these stressors remain understudied in large predatory fishes, including sharks. We tested for behavioural and physiological responses of blacktip reef shark (Carcharhinus melanopterus) neonates to climate change relevant changes in temperature (28 and 31 °C) and carbon dioxide partial pressures (pCO2; 650 and 1050 µatm) using a fully factorial design. Behavioural assays (lateralisation, activity level) were conducted upon 7–13 days of acclimation, and physiological assays (hypoxia tolerance, oxygen uptake rates, acid–base and haematological status) were conducted upon 14–17 days of acclimation. Haematocrit was higher in sharks acclimated to 31 °C than to 28 °C. Significant treatment effects were also detected for blood lactate and minimum oxygen uptake rate; although, these observations were not supported by adequate statistical power. Inter-individual variability was considerable for all measured traits, except for haematocrit. Moving forward, studies on similarly 'hard-to-study' species may account for large inter-individual variability by increasing replication, testing larger, yet ecologically relevant, differences in temperature and pCO2, and reducing measurement error. Robust experimental studies on elasmobranchs are critical to meaningfully assess the threat of global change stressors in these data-deficient species