Fish Physiology and Biochemistry

The in vitro hepatic 5′-monodeiodination of thyroxine (T4) to triiodothyronine (T3) in Oreochromis mossambicus, Channa striata, Clarias batrachus, Cyprinus carpio and Oxyeleotris marmorata was found to be time, pH and temperature dependent, and related to the amount of substrate (T4) and homogenate introduced into the reaction vessel, in a manner which was consistent with Menton-Michaelis kinetics, and thus indicative of an enzyme-regulated process. Dithiothreitol introduced into the reaction vessel stimulated T3 production in a dose-related manner. Hepatic 5′-monodeiodinase activity was also detected in a further 28 species of teleosts suggesting that the peripheral monodeiodination of T4, which is well-documented in salmonids, is also widespread amongst other teleost fishes. All species examined exhibited evidence of enzymatic deiodination, but there were marked differences in Km and Vmax values between the species. There was no apparent phylogenetic or environmental relationships to explain the widely divergent Km and/or Vmax values, nor was there a correlation between Km and Vmax when the species were considered together.

The acute effects of 3 fish anaesthetics (MS222, metomidate and AQUI-S) were investigated on 3 parts of the cardiovascular system of Chinook salmon (Oncorhynchus tshawytscha). All 3 anaesthetics reduced the contractile force of paced strips of ventricular myocardium. MS222 reduced the contractility by almost 75%, and was more potent than metomidate and AQUI-S, which reduced the contractility by about 25%. MS222 blocked vagal nerve transmission to the heart at the normally applied anaesthetic concentration (NAAC) for Chinook salmon, whereas metomidate and AQUI-S required 100 times their NAACs to have the same effect. Using myography, MS222 and AQUI-S caused a maximal 30–40% dilation of EBAs at 10% NAAC, whereas the equivalent effect with metomidate was only seen at 100% NAAC. MS222 again caused the greatest dilation of the ABAs. AQUI-S dilated the ABAs at up to 50% NAAC, but this was reversed so that there was no dilation at 100% NAAC. Metomidate did not affect the ABAs. These data from in vitro and in situ experiments, which generally show inhibitory effects, are used to suggest possible cardiovascular outcomes in anaesthetised Chinook salmon.

Although Pacific bluefin tuna is a species that relies on vision, its photopic visual function is not well known; we therefore recorded electroretinograms to investigate photopic spectral sensitivity in juveniles of this species (49–81 days post-hatch; standard length 74–223 mm). The peak spectral sensitivity wavelength was 505 nm. We estimated that two (λmax = 512–515 nm and 423–436 nm) or three (λmax = 512–515 nm, 423–436 nm, and 473 nm) types of cone visual pigments contribute to photopic vision; these spectral sensitivities are adapted to surface water habitats in clear ocean and coastal water.

The energy density (ED) of nine species of sub-Antarctic fishes was estimated by calorimetry. The fish, seven notothenioids, one atherinopsid and one galaxiid, represents some of the more abundant species in the ichthyofauna of the Beagle Channel. Principal-components analysis (PCA) of the ED of the different organs/tissues indicated that PC1 and PC2 accounted for 87% of the variability. Separation along PC1 corresponded to differences in muscle and liver energy densities whereas separation along PC2 corresponded to differences in the ED of the gonads. Differences between species were significant except for P. sima. Inclusion of the gonadosomatic index (GSI) as an explanatory variable enabled us to establish the existence of energy transfer from muscle and liver to the gonads in ripe P. tessellata females. Total ED values varied between 4.21 and 6.26 kJ g−1, the pelagic Odontesthes sp. being the species with the highest ED. A significant relationship between ED and muscle dry weight (DWM) was found for all the species except P. tessellata. These data are the first direct estimates of ED of fishes from the Beagle Channel.

The efficiency of enzymatic conversion of glucose into glucose-6-phosphate in fish fed carbohydrates is still a matter of debate. This study reports the first ever identification of hexokinase-family members at a molecular level in fish. The deduced amino acid sequences of six cDNA clones allowed their assignement either to hexokinase IV (glucokinase) for rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio) and gilthead seabream (Sparus aurata) or to hexokinase I/II for the common carp (two clones) and the gilthead seabream, based on their similarities to known members of the HK family. The evolutionary relationships among these sequences were investigated by phylogenetic analysis. These hexokinase sequences constitute potential molecular tools for analysis of interspecific differences in dietary carbohydrate utilization in fish.

Juvenile tilapias (Oreochromis niloticus × O. aureus), with average body weights of groups I, II and III of 55.14, 122.82, and 225.68 g respectively were used to study changes in the activity of protease, amylase, and lipase in different organs and sections of the alimentary tract. Additionally, tract containing digesta, which is closer to physiological conditions, may help reveal the mechanisms of whole chemical digestion process. Another group of tilapias (mean body weight of 111.81 ± 0.14 g) was used to analyze changes in activity of protease, lipase, and amylase in response to varying pH and temperatures. The activity of enzymes in all digestive organs increased when body weight increased from 55 to 122 g, and the activity further increased when the weights increased from 122 to 225 g. The rates of increase in digestive enzyme activity were significantly greater than the rates of growth of the fish. When the activity was compared in different sections of the tract, a similar regularity was found for the three enzymes in all fish. The highest amounts of activity were seen in the foregut and the lowest in the rectum, with the middle amount of activity in the hindgut. The pH ranges for the maximum activity of tilapia protease in the stomach and intestine were 1.6–3.3 and 9.5–10.5 respectively and the optimal temperature in these organs was 55°C. The pH ranges for the maximum amylase activity in the intestine and hepatopancreas were 6–7 and 7.5 respectively, and high amylase activity in these organs was found at 25–35°C. The activity of lipase in the intestine remained unchanged at pH 6.0–9.0 and with temperatures between 25 and 35°C. These results suggest that the ability of tilapia to utilize feed varies at different growth stages and remains relatively stable with changes in environmental pH and temperature, which should be taken into account when designing feed formulas for tilapia.

The regulation mechanism of the hybrid yellow catfish “Huangyou-1” was assessed under conditions of hypoxia and reoxygenation by examination of oxygen sensors and by monitoring respiratory metabolism, oxidative stress, and apoptosis. The expressions of genes related to oxygen sensors (HIF-1α, HIF-2α, VHL, HIF-1β, PHD2, and FIH-1) were upregulated in the brain and liver during hypoxia, and recovered compared with control upon reoxygenation. The expressions of genes related to glycolysis (HK1, PGK1, PGAM2, PFK, and LDH) were increased during hypoxia and then recovered compared with control upon reoxygenation. The mRNA levels of CS did not change during hypoxia in the brain and liver, but increased during reoxygenation. The mRNA levels of SDH decreased significantly only in the liver during hypoxia, but later increased compared with control upon reoxygenation in both tissues. Under hypoxic conditions, the expressions of genes related to oxidative stress (SOD1, SOD2, GSH-Px, and CAT) and the activity of antioxidant enzymes (SOD, CAT, and GSH-Px) and MDA were upregulated compared with control. The expressions of genes related to apoptosis (Apaf-1, Bax, Caspase 3, Caspase 9, and p53) were higher than those in control during hypoxic exposure, while the expressions of Bcl-2 and Cyt C were decreased. The findings of the transcriptional analyses will provide insights into the molecular mechanisms of hybrid yellow catfish “Huangyou-1” under conditions of hypoxia and reoxygenation. Overall, these findings showed that oxygen sensors of “Huangyou-1” are potentially useful biomarkers of environmental hypoxic exposure. Together with genes related to respiratory metabolism, oxidative stress and apoptosis occupy a quite high position in enhancing hypoxia tolerance. Our findings provided new insights into the molecular regulatory mechanism of hypoxia in “Huangyou-1.”