The importance of the crocodilian hepatic piston pump to ventilation during altered respiratory demand

Crocodilians ventilate through a combination of the intercostal muscles, the abdominal muscles and the diaphragmaticus muscle. Previous studies report that the caudal-cranial movement of the liver during the ventilatory cycle by the diaphragmaticus muscle, termed the hepatic piston pump, is solely responsible for ventilation in floating caimans. However, the importance of the hepatic-piston pump to ventilation in crocodilians under altered conditions of ventilatory demand is unknown. The hepatic-piston pump made only a limited contribution to ventilation while crocodiles rested at 30OC, following a decrease in temperature (20OC, reduced ventilatory demand), and during hypercapnia (5% CO2, increased ventilatory drive). The diaphragmatic muscle was important for facilitating ventilation during exercise (increased ventilatory demand) as loss of the hepatic piston pump, following inactivation of the diaphragmaticus muscle, compromised exercise induced increases in tidal volume and minute ventilation. A relative hyperventilation was induced by exercise (both with and without a functional hepatic piston pump) and, as a result, the alterations in ventilation following inactivation of the diaphragmaticus muscle did not significantly alter arterial oxygenation

Calibration of the HemoCue point-of-care analyser for determining haemoglobin concentration in a lizard and a fish

Haemoglobin concentration ([Hb]) is measured for a wide variety of animal studies. The use of point-of-care devices, such as the HemoCue, is becoming increasingly common because of their portability, relative ease of use and low cost. In this study, we aimed to determine whether the [Hb] of blue-tongued skink (Tiliqua nigrolutea) blood can be determined accurately using the HemoCue and whether the HemoCue overestimates the [Hb] of reptile blood in a similar manner to fish blood. Additionally, we aimed to test whether ploidy affected [Hb] determined by the HemoCue using blood from diploid and triploid Atlantic salmon (Salmo salar). The HemoCue Hb 201⁺ systematically overestimated [Hb] in both blue-tongued skinks and Atlantic salmon, and there was no difference between calibration equations determined for diploid or triploid salmon. The overestimation was systematic in both species and, as such, [Hb] determined by the HemoCue can be corrected using appropriate calibration equations

The accessory role of the diaphragmaticus muscle in lung ventilation in the estuarine crocodile Crocodylus porosus

Crocodilians use a combination of three muscular mechanisms to effect lung ventilation: the intercostal muscles producing thoracic movement, the abdominal muscles producing pelvic rotation and gastralial translation, and the diaphragmaticus muscle producing visceral displacement. Earlier studies suggested that the diaphragmaticus is a primary muscle of inspiration in crocodilians, but direct measurements of the diaphragmatic contribution to lung ventilation and gas exchange have not been made to date. In this study, ventilation, metabolic rate and arterial blood gases were measured from juvenile estuarine crocodiles under three conditions: (i) while resting at 30°C and 20°C; (ii) while breathing hypercapnic gases; and (iii) during immediate recovery from treadmill exercise. The relative contribution of the diaphragmaticus was then determined by obtaining measurements before and after transection of the muscle. The diaphragmaticus was found to make only a limited contribution to lung ventilation while crocodiles were resting at 30°C and 20°C, and during increased respiratory drive induced by hypercapnic gas. However, the diaphragmaticus muscle was found to play a significant role in facilitating a higher rate of inspiratory airflow in response to exercise. Transection of the diaphragmaticus decreased the exercise-induced increase in the rate of inspiration (with no compensatory increases in the duration of inspiration), thus compromising the exercise-induced increases in tidal volume and minute ventilation. These results suggest that, in C. porosus, costal ventilation alone is able to support metabolic demands at rest, and the diaphragmaticus is largely an accessory muscle used at times of elevated metabolic demand

Negligible differences in metabolism and thermal tolerance between diploid and triploid Atlantic salmon (Salmo salar)

The mechanisms that underlie thermal tolerance in aquatic ectotherms remain unresolved. Triploid fish have been reported to exhibit lower thermal tolerance than diploids, offering a potential model organism to better understand the physiological drivers of thermal tolerance. Here, we compared triploid and diploid juvenile Atlantic salmon (Salmo salar) in freshwater to investigate the proposed link between aerobic capacity and thermal tolerance. We measured specific growth rates (SGR) and resting (aerobic) metabolic rates (RMR) in freshwater at 3, 7 and 9 weeks of acclimation to either 10, 14 or 18°C. Additionally, maximum metabolic rates (MMR) were measured at 3 and 7 weeks of acclimation, and critical thermal maxima (CTmax) were measured at 9 weeks. Mass, SGR, and RMR differed between ploidies across all temperatures at the beginning of the acclimation period, but all three metrics converged between ploidies by week 7. Aerobic scope (MMR – RMR) remained consistent across ploidies, acclimation temperatures, and time. At 9 weeks, CTmax was independent of ploidy, but correlated positively with acclimation temperature despite the similar aerobic scope between acclimation groups. Our findings suggest that acute thermal tolerance is not modulated by aerobic scope, and the altered genome of triploid Atlantic salmon does not translate to reduced thermal tolerance of juvenile fish in freshwater

Physiological effects of dissolved oxygen are stage-specific in incubating Atlantic salmon (Salmo salar)

Oxygen availability is highly variable during salmonid incubation in natural redds and also in aquaculture incubation systems. Hypoxia generally decreases growth and aerobic metabolism prior to hatching, in parallel with eliciting physiological modifications that enhance oxygen delivery. However, it is less-well known whether developmental hyperoxia can drive the opposite effect. Moreover, there is insufficient understanding of stage-specific developmental windows during which ambient oxygen availability may be of greater or lesser impact to incubating embryos. Here, we tested the effects of hypoxia (50% dissolved oxygen: DO, % air saturation) and hyperoxia (150% DO) on the growth, routine aerobic metabolism (ṀO2rout) and hypoxia tolerance (O2crit) of Atlantic salmon (Salmo salar) during seven developmental windows throughout incubation. Embryos exposed to hyperoxia (150% DO) did not differ from the normoxic group in growth, ṀO2rout or O2crit at any developmental window. In contrast, embryos exposed to hypoxia grew slower and had a lower ṀO2rout, but had higher hypoxia tolerance (lower O2crit) than normoxic and hyperoxic counterparts. Interestingly, these differences were only apparent when the embryos were measured prior to hatching. Larvae (alevins) incubated in hypoxia following hatching grew similarly to normoxia-incubated alevins. Our results provide evidence that Atlantic salmon embryos are most sensitive to hypoxia prior to hatching, probably due to increasing (absolute) oxygen requirements concurrent with restricted oxygen diffusion through the egg. Moreover, the similarities between normoxia- and hyperoxia-incubated salmon demonstrate that embryos are not oxygen-limited under normoxic conditions

Hypoxia during incubation does not affect aerobic performance or haematology of Atlantic salmon (Salmo salar) when re-exposed in later life

Hypoxia in aquatic ecosystems is becoming increasingly prevalent, potentially reducing fish performance and survival by limiting the oxygen available for aerobic activities. Hypoxia is a challenge for conserving and managing fish populations and demands a better understanding of the short- and long-term impacts of hypoxic environments on fish performance. Fish acclimate to hypoxia via a variety of short- and long-term physiological modifications in an attempt to maintain aerobic performance. In particular, hypoxia exposure during early development may result in enduring cardio-respiratory modifications that affect future hypoxia acclimation capacity, yet this possibility remains poorly investigated. We incubated Atlantic salmon (Salmo salar) in normoxia (∼100% dissolved oxygen [DO, as percent air saturation]), moderate hypoxia (∼63% DO) or cyclical hypoxia (100-25% DO daily) from fertilization until 113 days post-fertilization prior to rearing all groups in normoxia for a further 8 months. At ∼11 months of age, subsets of each group were acclimated to hypoxia (50% DO) for up to 44 days prior to haematology, aerobic metabolic rate and hypoxia tolerance measurements. Hypoxia exposure during incubation (fertilization to 113 days post-fertilization) did not affect the haematology, aerobic performance or hypoxia tolerance of juvenile salmon in later life. Juveniles acclimated to hypoxia increased maximum aerobic metabolic rate and aerobic scope by ∼23 and ∼52%, respectively, when measured at 50% DO but not at 100% DO. Hypoxia-incubated juveniles also increased haematocrit and haemoglobin concentration but did not affect acute hypoxia tolerance (critical oxygen level and DO at LOE). Thus, while Atlantic salmon possess a considerable capacity to physiologically acclimate to hypoxia by improving aerobic performance in low oxygen conditions, we found no evidence that this capacity is influenced by early-life hypoxia exposure