The physiological consequences of a very large natural meal in a voracious marine fish, the staghorn sculpin (Leptocottus armatus).

Little information exists on physiological consequences when wild fish eat natural food. Staghorn sculpins at 10-13°C voluntarily consumed 15.8% of their body mass in anchovies. Gastric clearance was slow with >60% of the meal retained in the stomach at 48 h, and was not complete until 84 h. At 14-24 h post-feeding, pH was depressed by 3 units and Cl- concentration was elevated 2-fold in gastric chyme, reflecting HCl secretion, while in all sections of the intestine, pH declined by 1 pH unit but Cl- concentration remained unchanged. PCO2 and total ammonia concentration were greatly elevated throughout the tract, whereas PNH3 and HCO3- concentration were depressed. Intestinal HCO3- secretion rates, measured in gut sacs in vitro, were also lower in fed fish. Whole-animal O2 consumption rate was elevated approximately 2-fold for 72 h post-feeding, reflecting 'specific dynamic action', whereas ammonia and urea-N excretion rates were elevated about 5-fold. Arterial blood exhibited a modest 'alkaline tide' for about 48 h, but there was negligible excretion of metabolic base to the external seawater. PaCO2 and PaO2 remained unchanged. Plasma total amino acid concentration and total lipid concentration were elevated about 1.5-fold for at least 48 h, whereas small increases in plasma total ammonia concentration, PNH3 and urea-N concentration were quickly attenuated. Plasma glucose concentration remained unchanged. We conclude that despite the very large meal, slow processing with high efficiency minimizes internal physiological disturbances. This differs greatly from the picture provided by previous studies on aquacultured species using synthetic diets and/or force-feeding. Questions remain about the role of the gastro-intestinal microbiome in nitrogen and acid-base metabolism.

An atlas of plasma-accessible carbonic anhydrase availability in the model teleost, the rainbow trout.

The unique teleost oxygenation system that permits enhanced oxygen unloading during stress comprises three main characteristics: pH-sensitive haemoglobin, red blood cell (RBC) intracellular pH (pHi) protection, and a heterogeneous distribution of plasma-accessible carbonic anhydrase (paCA). A heterogeneous distribution of paCA is essential; its presence permits enhanced oxygen unloading during stress, while its absence at the gills maintains conditions for oxygen uptake by pH-sensitive haemoglobins. We hypothesised that paCA would be absent in all four gill arches, as has been previously indicated for arch two, and that paCA would be present in all other tissues. Through a suite of biochemical and molecular methods, we confirmed the absence of paCA from all four arches. We also found evidence for paCA in nine other tissues and a lack of paCA availability in the stomach. Expression was highly variable between tissues and suggests these differences may be associated with their respective metabolic activities. Additionally, we analysed the specific CA-IV isoform expressed within each tissue and showed almost complete separation of expression between tissues; CA-IVa was detected in the heart, brain, anterior intestine, and liver, whereas CA-IVb was detected in all intestine sections, pyloric caeca, kidney, and white muscle. This adds to a growing collection of work suggesting CA-IVa and b play divergent roles in gas exchange and ion/acid-base balance, respectively. The current study represents the first comprehensive atlas of paCA availability within the circulatory system of the model teleost, rainbow trout, and fills important gaps in our knowledge of this unique oxygenation system.

Do extreme postprandial levels of oxygen, carbon dioxide, and ammonia in the digestive tract equilibrate with the bloodstream in the freshwater rainbow trout (Oncorhynchus mykiss)?

The gastrointestinal tract (GIT) lumen of teleosts harbors extreme conditions, especially after feeding: high PCO2 (20-115 Torr), total ammonia (415-3710 µM), PNH3 (79-1760 µTorr in the intestine), and virtual anoxia (PO2 < 1 Torr). These levels could be dangerous if they were to equilibrate with the bloodstream. Thus, we investigated the potential equilibration of O2, CO2, and ammonia across the GIT epithelia in freshwater rainbow trout by monitoring postprandial arterial and venous blood gases in vivo and in situ. In vivo blood was sampled from the indwelling catheters in the dorsal aorta (DA) and subintestinal vein (SIV) draining the posterior intestine in the fasting state and at 4 to 48 h following catheter-feeding. To investigate possible ammonia absorption in the anterior part of the GIT, blood was sampled from the DA, SIV and hepatic portal vein (HPV) from anaesthetized fish in situ following voluntary feeding. We found minimal equilibration of all three gases between the GIT lumen and the SIV blood, with the latter maintaining pre-feeding levels (PO2 = 25-49 Torr, PCO2 = 6-8 Torr, and total ammonia = 117-134 µM and PNH3 = 13-30 µTorr at 48 h post-feeding). In contrast to the SIV, we found that the HPV total ammonia more than doubled 24 h after feeding (128 to 297 µM), indicative of absorption in the anterior GIT. Overall, the GIT epithelia of trout, although specialized for absorption, prevent dangerous levels of PO2, PCO2 and ammonia from equilibrating with the blood circulation.

Ammonia transport is independent of PNH3 gradients across the gastrointestinal epithelia of the rainbow trout: A role for the stomach.

Although the gastrointestinal tract (GIT) is an important site for nitrogen metabolism in teleosts, the mechanisms of ammonia absorption and transport remain to be elucidated. Both protein catabolism in the lumen and the metabolism of the GIT tissues produce ammonia which, in part, enters the portal blood through the anterior region of the GIT. The present study examined the possible roles of different GIT sections of rainbow trout (Oncorhynchus mykiss) in transporting ammonia in its unionized gas form-NH3 -by changing the PNH3 gradient across GIT epithelia using in vitro gut sac preparations. We also surveyed messenger RNA expression patterns of three of the identified Rh proteins (Rhbg, Rhcg1, and  Rhcg2) as potential NH3 transporters and NKCC as a potential ammonium ion (NH4 + ) transporter along the GIT of rainbow trout. We found that ammonia absorption is not dependent on the PNH3 gradient despite expression of Rhbg and Rhcg2 in the intestinal tissues, and Rhcg2 in the stomach. We detected no expression of Rhbg in the stomach and no expression of Rhcg1 in any GIT tissues. There was also a lack of correlation between ammonia transport and [NH4 + ] gradient despite NKCC expression in all GIT tissues. Regardless of PNH3 gradients, the stomach showed the greatest absorption and net tissue consumption of ammonia. Overall, our findings suggest nitrogen metabolism zonation of GIT, with stomach serving as an important site for the absorption, handling and transport of ammonia that is independent of the PNH3 gradient.

Post-prandial respiratory gas and acid-base profiles in the gastrointestinal tract and its venous drainage in freshwater rainbow trout (Oncorhynchus mykiss) and seawater English sole (Parophrys vetulus).

The basic respiratory gas and acid-base conditions inside the lumen of the gastrointestinal tract (GIT) and blood draining the tract are largely unestablished in teleost fishes after feeding, though there have been some recent novel discoveries on freshwater rainbow trout (Oncorhynchus mykiss) and seawater English sole (Parophrys vetulus). The present study examined in greater detail the gas (PO2, PCO2, PNH3) and acid-base profiles (pH, [HCO3-], total [ammonia]) in the lumen of the stomach, the anterior, mid, and posterior intestine, as well as the venous drainage (subintestinal and/or hepatic portal vein) of the GIT in these two species 20 h post-feeding. Both species had high PCO2, PNH3, and total [ammonia], and low PO2 (virtual anoxia) in the lumens throughout all sections of the GIT, and high [HCO3-] in the intestine. Total [ammonia], PNH3, and [HCO3-] increased from anterior to posterior intestine in both species. P. vetulus had higher intestinal total [ammonia] and lower [HCO3-] than O. mykiss post feeding, but total [ammonia] was much higher in the stomach of O. mykiss. Despite the extreme conditions in the lumen, both arterial and venous blood showed relatively lower PCO2, total [ammonia] and higher PO2, implying limited equilibration between the two compartments. The higher [HCO3-] and lower total [ammonia] in the intestinal lumen of the freshwater O. mykiss than the seawater P. vetulus suggest the need for future comparative studies using conspecifics fed identical diets but acclimated to the two different salinities in order to understand the potential role of environmental salinity and associated osmoregulatory processes underlying these differences.

An in vitro study of urea and ammonia production and transport by the intestinal tract of fed and fasted rainbow trout: responses to luminal glutamine and ammonia loading.

Digestion of dietary protein in teleosts results in high ammonia levels within the intestinal chyme that may reach concentrations that are many-fold greater than blood plasma levels. We used in vitro gut sac preparations of the ammoniotelic rainbow trout (Oncorhynchus mykiss) to investigate the role of the intestine in producing and transporting ammonia and urea, with specific focus on feeding versus fasting, and on responses to loading of the lumen with 2 mmol L-1 glutamine or 2 mmol L-1 ammonia. Feeding increased not only ammonia production and both mucosal and serosal fluxes, but also increased urea production and serosal fluxes. Elevated urea production was accompanied by an increase in arginase activity but minimal CPS III activity, suggesting that urea may be produced by direct arginolysis. The ammonia production and serosal fluxes increased in fasted preparations with glutamine loading, indicating an ability of the intestinal tissue to deaminate glutamine and perhaps use it as an energy source. However, there was little evidence of urea production or transport resulting from the presence of glutamine. Furthermore, the intestinal tissues did not appear to convert surplus ammonia to urea as a detoxification mechanism, as urea production and serosal flux rates decreased in fed preparations, with minimal changes in fasted preparations. Nevertheless, there was indirect evidence of detoxification by another pathway, as ammonia production rate decreased with ammonia loading in fed preparations. Overall, our study suggests that intestinal tissues of rainbow trout have the ability to produce urea and detoxify ammonia, likely via arginolysis.

Reduced hypoxia tolerance and survival at elevated temperatures may limit the ability of Amazonian fishes to survive in a warming world.

The Amazon basin contains more than 20% of the world's freshwater fishes, many of ecological and economical importance. An increase in temperature of 2.2 to 7 °C is predicted to occur within the next century in the worst-case scenario of climate change predictions, which will likely be associated with an increase in the prevalence and duration of reduced water oxygen levels (hypoxia). Furthermore, there is an increasing frequency of heat waves in the Amazon basin, which exacerbates issues related to temperature and hypoxia. Increases in temperature and hypoxia both constrain an organism's ability to supply oxygen to metabolizing tissues, thus the ability to cope with thermal and hypoxic stress may be correlated. Here, we reveal a positive correlation between acute thermal tolerance and acute hypoxia tolerance amongst 37 Amazonian fish species at the current river temperatures of 28-31 °C. The effects of long-term (10 days or 4 weeks) increases in temperature were investigated in a subset of 13 species and demonstrated that 2 species failed to acclimate and survive at 33 °C, 9 species failed at 35 °C, and only 2 species survived up to 35 °C. Of those that survived long-term exposure to 33 or 35 °C, the majority of the species demonstrated only an improvement in acute thermal tolerance. In contrast, hypoxia tolerance was reduced following acute- and long-term exposure to 33, 35 or 37 °C in all species investigated. The results of this study suggest that many of the fish species that inhabit the Amazon may be at risk during both short- and long-term temperature increases and these risks are exacerbated by the associated environmental hypoxia.

The gaseous gastrointestinal tract of a seawater teleost, the English sole (Parophrys vetulus).

There has been considerable recent progress in understanding the respiratory physiology of the gastrointestinal tract (GIT) in teleosts, but the respiratory conditions inside the GIT remain largely unknown, particularly the luminal PCO2 and PO2 levels. The GIT of seawater teleosts is of special interest due to its additional function of water absorption linked to HCO3- secretion, a process that may raise luminal PCO2 levels. Direct measurements of GIT PCO2 and PO2 using micro-optodes in the English sole (Parophrys vetulus; anaesthetized, artificially ventilated, 10-12 °C) revealed extreme luminal gas levels. Luminal PCO2 was 14-17 mmHg in the stomach and intestinal segments of fasted sole, considerably higher than arterial blood levels of 5 mmHg. Moreover, feeding, which raised intestinal HCO3- concentration, also raised luminal PCO2 to 34-50 mmHg. All these values were higher than comparable measurements in freshwater teleosts, and also greater than environmental CO2 levels of concern in aquaculture or global change scenarios. The PCO2 values in subintestinal vein blood draining the GIT of fed fish (28 mmHg) suggested some degree of equilibration with high luminal PCO2, whereas subintestinal vein PO2 levels were relatively low (9 mmHg). All luminal sections of the GIT were virtually anoxic (PO2 ≤ 0.3 mmHg), in both fasted and fed animals, a novel finding in teleosts.

The effect of temperature acclimation on thermal tolerance, hypoxia tolerance and aerobic scope in two subspecies of sheepshead minnow; Cyprinodon variegatus variegatus and Cyprinodon variegatus hubbsi.

The freshwater teleost Cyprinodon variegatus hubbsi (Cvh) diverged from its euryhaline relative Cyprinodon variegatus variegatus (Cvv) ~150 kya and these subspecies are physiologically distinct in their osmoregulatory capabilities. Cvv inhabits intertidal estuaries and saltwater marshes along the Gulf of Mexico and Atlantic coast, where they experience a broad temperature range from -1.9 to 43 °C and frequent bouts of hypoxia. In contrast, Cvh lives in several lakes in central Florida, where temperature is more stable (12-31 °C) and hypoxia is uncommon. To assess whether relaxed selective pressure on Cvh has resulted in reduced temperature and hypoxia tolerance, a comparative study on the effects of acclimation to 25, 30 and 35 °C on critical thermal tolerance (CTMax), hypoxia tolerance, and aerobic scope was performed. The CTMax was similar between subspecies and positively correlated with acclimation temperature. Neither subspecies, however, could survive at 38 °C for a prolonged period of time. In general, Cvv displayed greater hypoxia tolerance and aerobic scope relative to Cvh over the range of acclimation temperatures. Routine metabolic rate was significantly lower while maximum metabolic rate and aerobic scope were significantly higher in Cvv, but only in fish acclimated to 30 °C. Overall, the different responses of Cvh to relaxed selective pressure suggest these traits are weakly linked physiologically in these fishes.