Biology subjects

Kwan, G. T.

Publications and source records attributed to Kwan, G. T..

9 recordsLinked to original sources

bioRxiv2024

Air Pocket for a Non-Air-Breathing Fish: Developing Respirometry Methodology for the critically endangered Delta Smelt

Respirometry quantifies O2 consumption to estimate metabolic activity across stressor(s), and the resulting data is central for devising conservation strategies for fishes. Due to its rapid adoption and widespread utility, protocols have been established to foster quality control and increase consistency. However, the application of established respirometry protocols on the Delta Smelt (Hypomesus transpacificus) have been largely unsuccessful, with reports over the past 12 years documenting high mortality rates that can only be circumvented with very short measurement duration ([~]4 h). This prevents the measurement of standard metabolic rate (SMR) and critical O2 tension (Pcrit), and ultimately obstructed physiological research, management actions and conservation tools development. In this study, we detailed the anecdotal, behavioral and empirical evidence that culminated in successful measurements of Delta Smelt SMR and Pcrit at 10, 12, 15, 17 and 19{degrees}C. We discovered the Delta Smelt is physotomous and requires frequent refilling of its air bladder to maintain buoyancy. Inspired by air-breathing fish respirometry, the inclusion of an air pocket within their chamber led to considerably more robust respirometry measurements compared to past attempts. The influence of fasting and time of day on O2 consumption were also examined. Altogether, this methodology paves the way for acquiring critical knowledge needed to inform Delta Smelt research and conservation (e.g. the development of metabolic index). The lessons learned may also have implications for other aspects of Delta Smelt conservation such as decreasing conservation aquaculture operating costs and improving survival during fish transport and supplementation.

physiology↗

bioRxiv2024

Gill ionocyte remodeling mediates blood pH regulation in rockfish (Sebastes diploproa) exposed to environmentally relevant hypercapnia

Marine fishes excrete excess H+ using basolateral Na+/K+-ATPase (NKA) and apical Na+/H+-exchanger 3 (NHE3) in gill ionocytes. However, the mechanisms that regulate H+ excretion during exposure to environmentally relevant hypercapnia (ERH) remain poorly understood. Here, we explored transcriptomic, proteomic, and cellular responses in gills of juvenile splitnose rockfish (Sebastes diploproa) exposed to three days of ERH conditions (pH [~]7.5; [~]1,600 atm pCO2). Blood pH was fully regulated at [~]7.75 despite a lack of significant changes in gill (1) mRNAs coding for proteins involved in blood acid-base regulation, (2) total NKA and NHE3 protein abundance, and (3) ionocyte density. However, ERH-exposed rockfish demonstrated increased NKA and NHE3 abundance on the ionocyte plasma membrane coupled with wider apical membranes and greater extension of apical microvilli. The observed gill ionocyte remodeling is consistent with enhanced H+ excretion that maintains blood pH homeostasis during exposure to ERH and does not necessitate changes at the expression or translation levels. These mechanisms of phenotypic plasticity may allow fishes to regulate blood pH during environmentally relevant acid-base challenges, and thus have important implications for both understanding how organisms respond to climate change and for selecting appropriate metrics to evaluate its impact on marine ecosystems.

physiology↗

bioRxiv2024

Diet and Size-at-Birth Affect Larval Rockfish Condition and Survival

Feeding success and maternal effects on larval size have long been hypothesized as important contributors to interannual recruitment variability in marine fishes. This study examined the feeding ecology and influences of diet and size-at-birth on length and growth of larval rockfishes (Sebastes spp.). Prey carbon biomass and selection were calculated from gut contents, size-at-birth was estimated using otolith core size, and recent growth was derived from outer otolith increment widths. Biomass contributions of preferred prey and otolith data were integrated into Bayesian hierarchical models predicting length and growth. Larvae primarily fed on and selected for copepod nauplii and Calanoid copepodites, modulating feeding with ontogeny and in response to prey availability. Based on carbon weight, the relative contribution of Calanoid copepodites to the diet was more strongly and positively correlated with length and growth than that of nauplii. Younger larvae experienced faster growth in association with Calanoid copepodite consumption than older larvae. Positive effects of core radius suggest that initial larval size, believed to be mediated by maternal provisioning, increases the likelihood of survival, larger size and faster growth. These findings ultimately provide evidence that selective feeding and size-at-birth mediate rockfish survival in early life stages.

ecology↗

bioRxiv2024

Fish Blood Response to Ash-Induced Environmental Alkalinization, and their Implications to Wildfire-Scarred Watersheds

Changes in land use, warming climate and increased drought have amplified wildfire frequency and magnitude globally. Ash mixing into aquatic systems after wildfires rapidly increases water pH, creating an additional threat to wildlife, especially species that are already threatened, endangered and/or migratory. Here, Chinook salmon (Oncorhynchus tshawytscha) yearlings acclimated to 15 or 20{degrees}C were exposed to an environmentally relevant concentration of ash (0.25% w/v) which caused water pH to rapidly rise from [~]8.1 to [~]9.2. Mortalities occurred within the first 12 hours, and was higher at the higher temperature (33 versus 20 %). The greatest differences in blood chemistry between the two temperatures were dramatically greater ([~]7.5-fold) and very rapid (within 1 hour) spikes in both plasma total ammonia (to [~]1200 {micro}M) and lactate (to [~]6 mM) in warm-acclimated salmon, whereas cold-acclimated salmon experienced a much smaller and gradual rise in plasma total ammonia. Salmon at both temperatures experienced extracellular and intracellular alkalosis within 1 hour that recovered within 24 hours, but the alkalosis was smaller in magnitude in fish at warmer temperature. Impacts on plasma ion concentrations were relatively mild and plasma glucose increased by 2- to 4-fold at both temperatures. Notably, the increase in plasma total ammonia in fish at the warmer temperature was far faster and much greater than those reported in previous studies exposing fish despite higher water pH (9.4-10.5) induced without using ash. This suggests that ash has physiological impacts that cannot be explained by high water pH alone which may relate to the complex mixture of metals and organic compounds also released from ash. This demonstrates post-wildfire ash input can induce lethal yet previously unexplored physiological disturbances in fish and highlights the complex interaction with warmer temperatures typical of wildfire-scarred landscapes.

physiology↗

bioRxiv2023

Trophic Efficiency Facilitates Larval Shortbelly Rockfish (Sebastes jordani) Development

Identifying the factors influencing fish recruitment is critical for fishery management, and failure to do so can have major ecological and economic consequences. Many hypotheses over the past century have been proposed, and the recently postulated Trophic Efficiency in Early Life (TEEL) hypothesis argues that a shorter food chain length can result in more efficient energy transfer from primary producers to young fishes, thereby increasing growth rate and larval condition, reducing early-life mortality and ultimately leading to a stronger recruitment cohort. To test this hypothesis, we analyzed the trophic position (TP) through compound-specific isotopic analysis of amino acids, as well as otolith microstructure and stomach content of larval shortbelly rockfish (Sebastes jordani). Results show larval rockfish that ate lower TP prey were both heavier and faster growing. This suggests the trophic characteristics of early life diet are critical to larval survival, and provide evidence in support of the TEEL hypothesis.

ecology↗

bioRxiv2023

Immunohistochemical and ultrastructural characterization of the inner ear epithelial cells of splitnose rockfish (Sebastes diploproa)

The inner ear of teleost fish regulates the ionic and acid-base chemistry and secretes the protein matrix of the endolymph to facilitate otolith biomineralization, which are used to maintain vestibular and auditory functions. The otolith is biomineralized in a concentric ring pattern corresponding to seasonal growth, and this CaCO3 polycrystal has become a vital aging and life-history tool for fishery managers, ecologists, and conservation biologists. Moreover, biomineralization patterns are sensitive to environmental variability including climate change, thereby threatening the accuracy and relevance of otolith-reliant toolkits. However, the cellular biology of the inner ear is poorly characterized, which is a hurdle for a mechanistic understanding of the underlying processes. This study provides a systematic characterization of the cell types in the inner ear of splitnose rockfish (Sebastes diploproa). Scanning electron microscopy revealed the apical morphologies of the six inner ear cell types. Additionally, immunostaining and confocal microscopy characterized the expression and subcellular localization of the proteins Na+/K+-ATPase, carbonic anhydrase, V-type H+-ATPase, Na+-K+-2Cl--Co-Transporter, Otolith Matrix Protein 1, and Otolin-1 in six inner ear cell types bordering the endolymph. This fundamental cytological characterization of the rockfish inner ear epithelium illustrates the intricate physiological processes involved in otolith biomineralization, and highlights how greater mechanistic understanding is necessary to predict their multi-stressor responses to future climate change.

physiology↗

bioRxiv2022

Optimizing immunostaining of archival fish samples to enhance museum collection potential

Immunohistochemistry (IHC) is a powerful biochemical technique that uses antibodies to specifically label and visualize proteins of interests within biological samples. However, fluid-preserved specimens within natural history collection often use fixatives and protocols that induce high background signal (autofluorescence), which hampers IHC as it produces low signal-to-noise ratio. Here, we explored techniques to reduce autofluorescence using sodium borohydride (SBH), citrate buffer, and their combination on fish tissue preserved with paraformaldehyde, formalin, ethanol, and glutaraldehyde. We found SBH was the most effective quenching technique, and applied this pretreatment to the gill or skin of 10 different archival fishes - including specimens that had been preserved in formalin or ethanol for up to 65 and 37 years, respectively. The enzyme Na+/K+-ATPase (NKA) was successfully immunostained and imaged using confocal fluorescence microscopy, allowing for the identification and characterization of NKA-rich ionocytes essential for fish ionic and acid-base homeostasis. Altogether, our SBH-based method facilitates the use of IHC on archival samples, and unlocks the historical record on fish biological responses to environmental factors (such as climate change) using specimens from natural history collections that were preserved decades to centuries ago. HighlightsO_LISodium borohydride pretreatment reduced aldehyde-induced autofluorescence C_LIO_LISuccessfully immunostained archival samples of various fixative and fixation time C_LIO_LILarval fish that was formalin-fixed for 63-65 years was successfully immunostained C_LI

biochemistry↗

bioRxiv2021

Elucidating the acid-base mechanisms underlying otolith overgrowth in fish exposed to ocean acidification

Over a decade ago, ocean acidification (OA) exposure was reported to induce otolith overgrowth in teleost fish. This phenomenon was subsequently confirmed in multiple species; however, the underlying physiological causes remain unknown. Here, we report that splitnose rockfish (Sebastes diploproa) exposed to ~1,600 atm pCO2 (pH ~7.5) were able to fully regulated the pH of both blood and endolymph (the fluid that surrounds the otolith within the inner ear). However, while blood was regulated around pH 7.80, the endolymph was regulated around pH ~8.30. These different pH setpoints result in increased pCO2 diffusion into the endolymph, which in turn leads to proportional increases in endolymph [HCO3-] and [CO32-]. Endolymph pH regulation despite the increased pCO2 suggests enhanced H+ removal. However, a lack of differences in inner ear bulk and cell-specific Na+/K+-ATPase and vacuolar type H+-ATPase protein abundance localization pointed out to activation of preexisting ATPases, non-bicarbonate pH buffering, or both, as the mechanism for endolymph pH-regulation. These results provide the first direct evidence showcasing the acid-base chemistry of the endolymph of OA-exposed fish favors otolith overgrowth, and suggests that this phenomenon will be more pronounced in species that count with more robust blood and endolymph pH regulatory mechanisms.

physiology↗

bioRxiv2021

Rapid blood acid-base regulation by European sea bass (Dicentrarchus labrax) in response to sudden exposure to high environmental CO2

Fish in coastal ecosystems can be exposed to acute variations in CO2 that can approach 1 kPa CO2 (10,000 atm). Coping with this environmental challenge will depend on the ability to rapidly compensate the internal acid-base disturbance caused by sudden exposure to high environmental CO2 (blood and tissue acidosis); however, studies about the speed of acid-base regulatory responses in marine fish are scarce. We observed that upon exposure to ~1 kPa CO2, European sea bass (Dicentrarchus labrax) completely regulate erythrocyte intracellular pH within ~40 minutes, thus restoring haemoglobin-O2 affinity to pre-exposure levels. Moreover, blood pH returned to normal levels within ~2 hours, which is one of the fastest acid-base recoveries documented in any fish. This was achieved via a large upregulation of net acid excretion and accumulation of HCO3- in blood, which increased from ~4 to ~22 mM. While the abundance and intracellular localisation of gill Na+/K+-ATPase (NKA) and Na+/H+ exchanger 3 (NHE3) remained unchanged, the apical surface area of acid-excreting gill ionocytes doubled. This constitutes a novel mechanism for rapidly increasing acid excretion during sudden blood acidosis. Rapid acid-base regulation was completely prevented when the same high CO2 exposure occurred in seawater with experimentally reduced HCO3- and pH, likely because reduced environmental pH inhibited gill H+ excretion via NHE3. The rapid and robust acid-base regulatory responses identified will enable European sea bass to maintain physiological performance during large and sudden CO2 fluctuations that naturally occur in coastal environments. Summary statementEuropean sea bass exposed to 1 kPa (10,000 atm) CO2 regulate blood and red cell pH within 2 hours and 40 minutes, respectively, protecting O2 transport capacity, via enhanced gill acid excretion.

physiology↗