Altered brain ion gradients following compensation for elevated CO2 are linked to behavioural alterations in a coral reef fish

Neurosensory and behavioural disruptions are some of the most consistently reported responses upon exposure to ocean acidification-relevant CO2 levels, especially in coral reef fishes. The underlying cause of these disruptions is thought to be altered current across the GABAA receptor in neuronal cells due to changes in ion gradients (HCO3− and/or Cl−) that occur in the body following compensation for elevated ambient CO2. Despite these widely-documented behavioural disruptions, the present study is the first to pair a behavioural assay with measurements of relevant intracellular and extracellular acid-base parameters in a coral reef fish exposed to elevated CO2. Spiny damselfish (Acanthochromis polyacanthus) exposed to 1900 μatm CO2 for 4 days exhibited significantly increased intracellular and extracellular HCO3− concentrations and elevated brain pHi compared to control fish, providing evidence of CO2 compensation. As expected, high CO2 exposed damselfish spent significantly more time in a chemical alarm cue (CAC) than control fish, supporting a potential link between behavioural disruption and CO2 compensation. Using HCO3− measurements from the damselfish, the reversal potential for GABAA (EGABA) was calculated, illustrating that biophysical properties of the brain during CO2 compensation could change GABAA receptor function and account for the behavioural disturbances noted during exposure to elevated CO2

Mapping QTL for sex and growth traits in Salt-Tolerant Tilapia (Oreochromis spp. X O. mossambicus)

In aquaculture, growth and sex are economically important traits. To accelerate genetic improvement in increasing growth in salt-tolerant tilapia, we conducted QTL mapping for growth traits and sex with an F2 family, including 522 offspring and two parents. We used 144 polymorphic microsatellites evenly covering the genome of tilapia to genotype the family. QTL analyses were carried out using interval mapping for all individuals, males and females in the family, respectively. Using all individuals, three suggestive QTL for body weight, body length and body thickness respectively were detected in LG20, LG22 and LG12 and explained 2.4% to 3.1% of phenotypic variance (PV). When considering only males, five QTL for body weight were detected on five LGs, and explained 4.1 to 6.3% of PV. Using only females from the F2 family, three QTL for body weight were detected on LG1, LG6 and LG8, and explained 7.9–14.3% of PV. The QTL for body weight in males and females were located in different LGs, suggesting that in salt-tolerant tilapia, different set of genes ‘switches’ control the growth in males and females. QTL for sex were mapped on LG1 and LG22, indicating multigene sex determination in the salt-tolerant tilapia. This study provides new insights on the locations and effects of QTL for growth traits and sex, and sets the foundation for fine mapping for future marker-assisted selection for growth and sex in salt-tolerant tilapia aquaculture

Exposure to boat noise in the field yields minimal stress response in wild reef fish

Aquatic anthropogenic noise is on the rise, with growing concern about its impact on species that are sensitive to low-frequency sounds (e.g. most fish and invertebrates). We investigated whether the reef fish Halichoeres bivittatus living in both noisy and quiet areas had differing levels of baseline stress (measured as whole-body cortisol) and whether they would exhibit a physiological stress response when exposed to boat noise playbacks. While the playback experiments significantly increased cortisol levels in fish from our experiment compared to baseline levels, there were minimal pairwise differences across treatments and no difference in baseline stress for fish living in noisy vs. quiet areas. These results may be explained by low overall auditory sensitivity, habituation to a fairly noisy environment (due to biological sounds), or that boat noise simply may not represent an immediate threat to survival in this species. These findings contrast recent studies that have shown elevated stress responses in fishes when exposed to boat noise and highlights that inter-specific differences must be considered when evaluating potential impacts of anthropogenic noise on marine life

Exposure to boat noise in the field yields minimal stress response in wild reef fish

Aquatic anthropogenic noise is on the rise, with growing concern about its impact on species that are sensitive to low-frequency sounds (e.g. most fish and invertebrates). We investigated whether the reef fish Halichoeres bivittatus living in both noisy and quiet areas had differing levels of baseline stress (measured as whole-body cortisol) and whether they would exhibit a physiological stress response when exposed to boat noise playbacks. While the playback experiments significantly increased cortisol levels in fish from our experiment compared to baseline levels, there were minimal pairwise differences across treatments and no difference in baseline stress for fish living in noisy vs. quiet areas. These results may be explained by low overall auditory sensitivity, habituation to a fairly noisy environment (due to biological sounds), or that boat noise simply may not represent an immediate threat to survival in this species. These findings contrast recent studies that have shown elevated stress responses in fishes when exposed to boat noise and highlights that inter-specific differences must be considered when evaluating potential impacts of anthropogenic noise on marine life

Beyond buoyancy and vision: the potential for the Root effect to deliver oxygen to tissues other than the swim bladder and eye

Teleost fish possess a unique, pH-sensitive hemoglobin (Hb) that, in the presence of an acidosis, substantially reduces the affinity and carrying capacity for O₂ (Root effect). To date, this efficient O₂ delivery mechanism is only known for filling a swim bladder (SB) against huge pressure gradients (> 50 atm) associated with depth and for oxygenating the metabolically active, yet avascular retinal tissue of the eye. In spite of the clear benefits to O₂ delivery for buoyancy and vision, no study has been conducted to determine whether the Root effect may be important in optimizing O₂ delivery to other tissues such as muscle, which is the focus of this research.During environmental or exercise-induced stress, blood pH may fall; however, some fish regulate red blood cell (RBC) intracellular pH (pHi) by releasing catecholamines that activate the sodium/proton (Na⁺/H⁺) exchanger (βNHE) on the RBC membrane. The βNHE removes H⁺s from the RBC resulting in an intracellular alkalosis, an increase in Hb–O₂ affinity, and O₂ uptake at the respiratory surfaces is safeguarded, which is the ultimate goal of this mechanism. In our proposed model, when adrenergically stimulated blood encounters plasma-accessible carbonic anhydrase (CA), an enzyme found in the RBC but also membrane-bound and potentially plasma-accessible in select locations, it will catalyze H⁺s removed from the RBC to form CO₂. This CO₂ will back-diffuse into the RBC creating an intracellular acidosis (extracellular alkalosis), reducing Hb–O₂ affinity, and ultimately elevating PO₂ via the Root effect. We created an in vitro closed system using rainbow trout (Oncorhynchus mykiss) blood where we can (1) simulate an acid-induced Root effect, (2) adrenergically stimulate the RBCs, and finally (3) short-circuit the βNHE via CA (CA-mediated Root effect), all of which can be monitored in real-time ( Fig. 1). Data generated currently support our Hypothesis: adrenergic RBC pH regulation can be short-circuited in the presence of plasma-accessible CA, therefore generating a Root effect increase in PO₂. In fact, if this scenario also occurs in the tissues of O. mykiss, CA-mediated short-circuiting of adrenergic pH regulation can facilitate an increase in PO₂ over 30 times that which would be generated in vertebrates possessing only a Bohr shift! We are ready to test our model in vivo by implanting fiber-optic O₂ sensors in O. mykiss muscle while simulating environmental and exercise stress with and without CA blockers. Furthermore, even though CA is not found in general circulation, there are membrane-bound and potentially plasma-accessible isoforms in muscle endothelia, and research is underway to localize this enzyme to understand the relationship between location and function of the short-circuiting.\ud \ud Teleost fish, which are more numerous than all other vertebrates combined (terrestrial and aquatic), have evolved an extraordinary O₂ delivery mechanism, the Root effect, that allows O₂ delivery to the eye and to the SB, thus allowing efficient buoyancy regulation, which may be one of the most important factors responsible for the extensive adaptive radiation in teleost fishes. Therefore, it is particularly interesting that the Root effect has not yet been investigated for general O₂ delivery. If the Root effect can also facilitate general O₂ delivery in vivo, which our data currently support, this would help shed insight into how the Root effect was selected for prior to the evolution of the βNHE, choroid gland and retia of the eye, and the gas gland and rete mirabile associated with the SB

Gas transport and exchange: interaction between O2 and CO2 exchange

The interaction between O2 and CO2 in the general circulation of fish exists at the level of hemoglobin within the red blood cell, and is determined largely by the magnitude of the Bohr and Haldane effects. Assuming steady-state conditions, a Bohr–Haldane coefficient of 0.35–0.5 (0.5 × the respiratory quotient, RQ) is optimal for tissue O2 delivery (excluding the swimbladder and eye), and greater values may be important for CO2 excretion and acid–base homeostasis. Many teleosts possess a nonlinear Bohr–Haldane coefficient over the oxygen-equilibrium curve (OEC), which alters the nature of the interaction when different regions of the OEC are used for gas exchange. Recent in vivo experiments indicate that Bohr–Haldane coefficients close to RQ (typically 0.7–1.0) may play an important role in facilitating tissue O2 delivery in vivo likely due to the existence of large disequilibrium states in the blood

Developing in warm water: irregular colouration and patterns of a neonate elasmobranch

[Extract] Temperature can impact embryonic development in nearly all vertebrates. This may be particularly evident in ectotherms—especially embryos developing in eggs outside of the female. Until hatching, the eggs can be restricted to their local thermal environment. Temperature can affect development rates, and skeletal abnormalities and abnormal colouration and patterns (e.g., snakes; Vinegar 1974) can result from elevated temperatures. While perhaps not directly life-threatening, temperature-mediated changes in colouration and pattern development may impact biological fitness because patterns are often required for camouflage, to attract mates, and/or for deterring predators

Function and control of the fish secondary vascular system, a contrast to mammalian lymphatic systems

Teleost fishes and mammalian lineages diverged 400 million years ago, and environmental requirements (water versus air) have resulted in marked differences in cardiovascular function between fish and mammals. Suggestions that the fish secondary vascular system (SVS) could be used as a model for the mammalian lymphatic system should be taken with caution. Despite molecular markers indicating similar genetic origin, functions of the SVS in teleost fish are probably different from those of the mammalian lymphatic system. We determined that, in resting glass catfish (Kryptopterus bicirrhis), plasma moves from the primary vascular system (PVS) to the SVS through small connecting vessels less than 10 m in diameter, smaller than the red blood cells (RBCs). During and following hypoxia or exercise, flow increases and RBCs enter the SVS, possibly via beta-adrenoreceptor-mediated dilation of the connecting vessels. The volume of the SVS can be large and, as RBCs flow into the SVS, the haematocrit of the PVS falls by as much as 50% of the resting value. Possible functions of the SVS, including skin respiration, ionic and osmotic buffering, and reductions in heart work and RBC turnover, are discussed

Heat shock protein (Hsp70) induced by a mild heat shock slightly moderates plasma osmolarity increases upon salinity transfer in rainbow trout (Oncorhynchus mykiss)

We have investigated whether mild heat shock, and resulting Hsp70 expression, can confer cross-protection against the stress associated with transfer from freshwater (FW) to seawater (SW) in juvenile rainbow trout (Oncorhynchus mykiss). In experimental Series I, juvenile trout reared in FW were transferred from 13.5 °C to 25.5 °C in FW, held for 2 h, returned to 13.5 °C for 12 h, and then transferred to 32 ppt SW at 13.5 °C. Branchial Hsp70 increased approximately 10-fold in the heat-shocked fish relative to the control by the end of recovery and remained high 2, 8, and 24 h post-salinity transfer. However, no clear differences could be detected in blood parameters (blood hemoglobin, hematocrit, MCHC, plasma Na⁺ and plasma osmolarity) or muscle water content between heat-shocked and sham-shocked fish in SW at any sampling interval (0, 2, 8, 24, 48, 120, 240 and 360 h post-SW transfer). In experimental Series II, trout acclimated to 8 °C were heat-shocked at 22 °C for 2 h, allowed to recover 18 h, and exposed to a more severe salinity transfer (either 36 or 45 ppt) than in Series I. Branchial Hsp70 levels increased approximately 6-fold in heat-shocked fish, but had declined to baseline after 120 h in SW. Plasma osmolarity and chloride increased in both groups upon transfer to 36 ppt; however, the increase was significantly less in heat-shocked fish when compared to the increase observed in sham-shocked fish at 24 h. No significant differences could be detected in branchial Na⁺/K⁺-ATPase activity or Na⁺/K⁺-ATPase α1a and α1b mRNA expression between the two groups. Our data indicate that a mild temperature shock has only modest effects on the ability of rainbow trout to resist osmotic stress during FW to SW transfer

Critical thermal maxima of early life stages of three tropical fishes: effects of rearing temperature and experimental heating rate

Marine ectotherms are often sensitive to thermal stress, and certain life stages can be particularly vulnerable (e. g., larvae or spawners). In this study, we investigated the critical thermal maxima (CTmax) of larval and early juvenile life stages of three tropical marine fishes (Acanthochromis polyacanthus, Amphiprion melanopus, and Lates calcarifer). We tested for potential effects of developmental acclimation, life stage, and experimental heating rates, and we measured metabolic enzyme activities from aerobic (citrate synthase, CS) and anaerobic pathways (lactate dehydrogenase, LDH). A slightly elevated rearing temperature neither influenced CTmax nor CS activity, which otherwise could have indicated thermal acclimation. However, we found CTmax to either remain stable (Acanthrochromis polyacanthus) or increase with body mass during early ontogeny (Amphiprion melanopus and Lates calcarifer). In all three species, faster heating rates lead to higher CTmax. Acute temperature stress did not change CS or LDH activities, suggesting that overall aerobic and anaerobic metabolism remained stable. Lates calcarifer, a catadromous species that migrates from oceanic to riverine habitats upon metamorphosis, had higher CTmax than the two coral reef fish species. We highlight that, for obtaining conservative estimates of a fish species' upper thermal limits, several developmental stages and body mass ranges should be examined. Moreover, upper thermal limits should be assessed using standardized heating rates. This will not only benefit comparative approaches but also aid in assessing geographic (re-) distributions and climate change sensitivity of marine fishes