Metabolic recovery in goldfish: A comparison of recovery from severe hypoxia exposure and exhaustive exercise

Severe hypoxia exposure and exhaustive exercise in goldfish both elicit a strong activation of substrate-level phosphorylation with the majority of the metabolic perturbations occurring in the white muscle. Approximately half of the muscle glycogen breakdown observed during severe hypoxia exposure was accounted for by ethanol production and loss to the environment, which limited the extent of muscle glycogen recovery when animals were returned to normoxic conditions. Ethanol production in goldfish is not solely a response to anoxia/hypoxia exposure however, as a transient increase in ethanol production was observed during the early stages of recovery from exhaustive exercise. These data suggest that ethanol production is a ubiquitous "anaerobic" end product, which accumulates whenever metabolic demands exceed mitochondrial oxidative potential. Exhaustive exercise and hypoxia exposure both caused a 7 to 8 micromol g(-1) wet mass increase in muscle [lactate] and the rates of recovery following these perturbations were similar. The rates of muscle PCr and pHi recovery after hypoxia exposure and exhaustive exercise were similar with levels returning to controls values within 0.5 h. Surprisingly, liver [glycogen] was not depleted during exposure to severe hypoxia, however, during recovery from both hypoxia and exercise dramatically different responses in liver [glycogen] were noted. During the early stages of recovery, liver [glycogen] transiently increased to high levels after exhaustive exercise, while during recovery from hypoxia there was a transient decrease in liver glycogen over the same time frame. Overall, this points to the liver playing a dramatically different role in facilitating recovery from exercise compared with hypoxia exposure.     

Distribution of aldehyde dehydrogenase and alcohol dehydrogenase in summer-acclimatized crucian carp, Carassius carassius L.

The tissue distribution of aldehyde dehydrogenase (ALDH) and alcohol dehydrogenase (ADH) in summer-acclimatized crucian carp showed almost the same exceptional pattern as previously found in winter-acclimatized specimens. There was a nearly complete spatial separation of ALDH and ADH; in other vertebrates these enzymes occur together. This exceptional enzyme distribution is probably an adaptation to the extraordinary ability of Carassius to produce ethanol as the major metabolic end product during anoxia. Since the crucian carp is less likely to encounter anoxia during the summer, the present results suggest that the crucian carp is unable to switch over to a 'normal' ALDH and ADH distribution in the summer. However, it is also possible that there is an advantage for the summer-acclimatized crucian carp in keeping ALDH and ADH separate, because of occasional anoxic periods.     

Chemical Alarm Signals Enhance Survival of Brook Charr (Salvelinus fontinalis) During Encounters with Predatory Chain Pickerel (Esox niger)

A diversity of aquatic organisms release chemical alarm signals when attacked or captured by a predator. These alarm signals are thought to warn other conspecifics of danger and, consequently, may benefit receivers by increasing their survival. Here we experimentally investigated the differences in behaviour and survival of hatchery-reared juvenile brook charr Salvelinus fontinalis that had been exposed to either brook charr skin extract (experimental treatment) or a control of swordtail skin extract (control treatment). Charr exposed to conspecific skin extract exhibited a significant reduction in movement and/or altered their foraging behaviour in the laboratory when compared with charr exposed to swordtail skin extract. We also exposed charr to either water conditioned by a single brook charr disturbed by a predatory bird model or water conditioned by a single undisturbed brook charr. Charr exposed to disturbance signals reduced activity significantly more than charr exposed to chemical stimuli from undisturbed charr. These results demonstrate the existence of both damage-released alarm signals and disturbance signals in brook charr. Wild brook charr also responded to damage-released alarm cues under natural conditions. Charr avoided areas of a stream with minnow traps labelled with conspecific alarm cues vs. control cues. During staged encounters with chain pickerel Esox niger in the laboratory, predator-naive charr fry were better able to evade the predator if they were previously warned by an alarm signal, thus suggesting a survival benefit to receivers. Collectively, these results demonstrate that the presence of alarm signals in brook charr has important implications for understanding predator–prey interactions.     

Seasonality of glycogen phosphorylase activity in crucian carp (<Emphasis Type="Italic">Carassius carassius</Emphasis> L<Emphasis Type="Italic">.</Emphasis>)

Seasonal changes in the activity of glycogen phosphorylase (GP), a rate-limiting enzyme of glycogen degradation, were examined in an anoxia-tolerant fish species, the crucian carp (Carassius carassius L.). In muscle and brain, the activity of GP remained constant throughout the year when tested at 25°C. In contrast, the activities of liver and heart GP displayed striking increases in summer. When seasonal temperature changes are taken into account, the activity of GP during the anoxic mid-winter is only 4–6% of its summer time activity in the muscle, heart and liver, and 13% in brain. In winter-acclimatized fish, experimental anoxia (1–6 weeks) caused sustained depression of the GP activity in heart and gills. In liver and muscle, a transient depression of GP activity occurred during the first week of anoxia but later GP activity recovered back to the normoxic level. GP of the brain was completely resistant to anoxia. In all studied tissues, the constitutive activity of GP is more than sufficient to degrade glycogen deposits during winter anoxia without anoxia-induced activation of GP. The seemingly paradoxical summer-time increase in the activity of liver and heart GP could be related to active life-style of the summer-acclimatized fish (growth, reproduction), the increased demand of energy and molecular precursors of anabolic metabolism being satisfied by preferential degradation of glycogen. The high glycogen content of winter-acclimatized crucian carp is not associated with the elevated GP activity or anoxic activation of GP.     

Seawater acclimation of diploid and triploid brook charr (Salvelinus fontinalis), diploid Arctic charr (Salvelinus alpinus), and their diploid and triploid hybrids

Seawater acclimation of diploid (FF) and triploid (F2F) brook charr Salvelinus fontinalis , diploid (AA) Arctic charr Salvelinus alpinus , and diploid (FA) and triploid (F2A) hybrids between female brook charr and male Arctic charr was investigated. Triploidization of brook charr and the hybrid did not have any effect on the acclimation. Seawater acclimation of the hybrid was achieved during the experimental period and was comparable to that observed in brook charr. Acclimation could not be ascertained in Arctic charr since the level of cortisol, a stress indicator, was still high at the end of the experiment. No relationship between either length or condition factors and plasma osmolality was observed. Elevated plasma cortisol concentrations in Arctic charr and in diploid or triploid hybrids, both in fresh water and sea water, indicate more favourable rearing conditions for brook charr.     

Control of carbohydrate metabolism in an anoxia-tolerant nervous system

Anoxia-tolerant animal models are crucial to understand protective mechanisms during low oxygen excursions. As glycogen is the main fermentable fuel supporting energy production during oxygen tension reduction, understanding glycogen metabolism can provide important insights about processes involved in anoxia survival. In this report we studied carbohydrate metabolism regulation in the central nervous system (CNS) of an anoxia-tolerant land snail during experimental anoxia exposure and subsequent reoxygenation. Glucose uptake, glycogen synthesis from glucose, and the key enzymes of glycogen metabolism, glycogen synthase (GS) and glycogen phosphorylase (GP), were analyzed. When exposed to anoxia, the nervous ganglia of the snail achieved a sustained glucose uptake and glycogen synthesis levels, which seems important to maintain neural homeostasis. However, the activities of GS and GP were reduced, indicating a possible metabolic depression in the CNS. During the aerobic recovery period, the enzyme activities returned to basal values. The possible strategies used by Megalobulimus abbreviatus CNS to survive anoxia are discussed.     

Influence of spring and summer water temperature on brook charr, Salvelinus fontinalis, growth and age structure in the Ford River, Michigan

Synopsis The influence of late spring and summer water temperatures on brook charr, Salvelinus fontinalis, growth and age structure was evaluated from 1984 to 1991 in the Ford River, Michigan. Temperature was monitored and brook charr sampled for vital statistics from late May through September using fyke nets and weirs at four locations within a 25.8 km section of stream. Scale analysis was used to determine captured brook charr age, past length at age and relative annual growth rates. Late spring and summer water temperature patterns varied between years with the greatest variability occurring in May and June. Age and size structure also varied between years and was significantly related to temperature. Years with cooler late spring and summer temperature patterns were dominated by older (age 2 and 3), larger brook charr, while years with warmer spring and summer temperature patterns were dominated by younger (age 1), smaller brook charr. Spring and summer temperature did not appear to have a significant effect on the growth of age 0 or age 1 brook charr. However, temperature was negatively related to brook charr growth from age 2 on. As spring and summer water temperatures are critical to brook charr growth and survival, it is important that a streams thermal regime be considered when establishing management goals for this species.     

Diel locomotor activity of brook charr, as determined by radiotelemetry

Locations by radiotelemetry during eight 24–h sampling periods in 1991, 1992, and 1993 were used to evaluate the diel activity patterns of adult brook charr Salvelinus fontinalis in two lakes of the Laurentian Shield (Québec, Canada). Based on the minimum distance travelled between two consecutive locations, adult brook charr were more active at dusk and at night than during the day. During daylight periods, individual fish displayed strong site selection, remaining in the same area along the shore from day to day. The fish left their inshore position at dusk, were highly mobile during the night, and returned to their focal point at sunrise, exhibiting diel homing behaviour. These results are supported by an increase in gillnet captures of adult brook charr in Lac Melchior at night. Data obtained by gillnet fishing in lakes Bondi and Simpson showed that juvenile (1+) brook charr were active throughout the diel cycle and that young-of-the-year were active mainly during the day, suggesting that there is an ontogenetic change in the diel activity of brook charr. Despite its limitations (restrictions on the number and size of tracked individuals, post-manipulation mortality, lifetime of transmitter battery), radiotelemetry allows for collection of multiple behavioural observations (distances travelled, instantaneous activity, site selection and homing) during the activity cycle of individual fish, and thus provides a finer degree of spatial resolution than alternative methods such as gillnetting.     

Complete amino acid sequence of rat liver alcohol dehydrogenase deduced from the cDNA sequence

Alcohol dehydrogenase (ADH) catalyzes the rate-determining reaction in the metabolism of ethanol. We report here the complete nucleotide sequence of a cDNA encoding rat liver ADH, and the deduced amino acid (aa) sequence of the protein. The rat enzyme contains a cluster of aa substitutions and an aa insertion in the region between aa residues 111 and 118, which is near the intron-exon junction reported for the human ADH gene. It also contains an additional cysteine in the highly variable region from aa residues 108-125 which may account for the unusual lability of rat ADH compared with ADH from other species.     

Evidence of anoxia-induced channel arrest in the brain of the goldfish (Carassius auratus)

The common goldfish (Carassius auratus) is extremely anoxia tolerant and here we provide evidence that "channel arrest" in the brain of these fish contributes to ATP conservation during periods of anoxia. Whole-cell patch-clamp recordings of slices taken from the telencephalon indicated that the N-methyl-d-aspartate (NMDA) receptor, an ionotropic glutamate receptor and Ca(2+)-channel, underwent a 40-50% reduction in activity during 40 min of acute anoxia. This is the first direct evidence of channel arrest in an anoxia-tolerant fish. Because goldfish produce ethanol as a byproduct of anaerobic metabolism we then conducted experiments to determine if the observed reduction in NMDA receptor current amplitude was due to inhibition by ethanol. NMDA receptor currents were not inhibited by ethanol (10 mmol L(-1)), suggesting that channel arrest of the receptor involved other mechanisms. Longer-term (48 h) in vivo exposure of goldfish to anoxic conditions (less than 1% dissolved O(2)) provided indirect evidence that a reduction in Na(+)/K(+)-ATPase activity also contributed to ATP conservation in the brain but not the gills. Anoxia under these conditions was characterized by a decrease in brain Na(+)/K(+)-ATPase activity of 30-40% by 24 h. Despite 90% reductions in the rates of ventilation, no change was observed in gill Na(+)/K(+)-ATPase activity during the 48-h anoxia exposure, suggesting that branchial ion permeability was unaffected. We conclude that rapid "channel arrest" of NMDA receptors likely prevents excitotoxicity in the brain of the goldfish, and that a more slowly developing decrease in Na(+)/K(+)-ATPase activity also contributes to the profound metabolic depression seen in these animals during oxygen starvation.     

The influence of groundwater withdrawal and land use changes on brook charr (<Emphasis Type="Italic">Salvelinus fontinalis</Emphasis>) thermal habitat in two coldwater tributaries in Michigan,U.S.A.

Brook charr (Salvelinus fontinalis) is a sentinel fish species which requires clean, cold water habitats. As such, many jurisdictions in the United States where brook charr are present use this species as an indicator of ecosystem health. In Michigan, groundwater-dominated streams are currently being impacted by increased groundwater withdrawal and land use/land cover changes which alter stream temperatures and their flow, and thus have the potential to significantly influence brook charr production and behavior. We quantified the influence of groundwater withdrawal and land use alteration on thermal habitat availability for brook charr using a groundwater modeling tool that estimated changes in baseflow to a stream segment based on changes to the groundwater system due to groundwater withdrawal and changes in rates of recharge related to landscape changes within a watershed. Projected stream temperature changes were calculated using a stream temperature modeling tool and compared to the range of temperature preferenda for brook charr in order to evaluate the potential impact of policy decisions regarding water extraction and land use/cover changes. The models predicted relatively small changes in both stream baseflow and consequently, stream temperature, with increased groundwater withdrawal rates. Land use/land cover alterations which we analyzed were shown to either mitigate or enhance the loss of brook charr thermal habitat as a result of groundwater withdrawal, depending on its relationship to recharge dynamics. This study emphasizes the importance of collaboration between water, land, and fisheries managers to ensure brook charr population viability, productivity, and sustainability in the face of environmental change, increasing water use and development in the watershed.     

Competitive interactions for foraging microhabitat among introduced brook charr, Salvelinus fontinalis, and native bull charr, S. confluentus, and westslope cutthroat trout, Oncorhynchus clarki lewisi, in a Montana stream

Competitive interactions for foraging microhabitat among introduced brook charr, Salvelinus fontinalis, and native bull charr, S. confluentus, and westslope cutthroat trout, Oncorhynchus clarki lewisi, were studied by species removal experiments in a tributary of the Flathead Lake and River system, northwestern Montana, focusing on brook charr influences on bull charr. When the three species were in sympatry, they interacted with each other, forming a size-structured, mixed-species dominance hierarchy in two stream pools. The influences of interference interactions were examined by measuring changes in five characteristics of foraging microhabitat and behavior, focal point height and velocity, cover use, and foraging rate and distance, after the successive removal of two species. Cutthroat trout removal resulted in increased foraging rates and distances, and decreased cover use for brook charr, but no changes for bull charr. After removal of brook charr from the two-species system, bull charr also increased foraging rates and distances and occupied more exposed positions. Moreover, total fish densities, which had initially decreased owing to the removal experiments, were partly compensated for by subsequent bull charr immigration, implying that competitive interactions with brook charr are an important factor in the mechanisms responsible for the regulation of bull charr densities, at least on a local scale.     

Habitat use and fish activity of landlocked Atlantic salmon and brook charr in a newly developed habitat compensation facility

Degradation and destruction of valuable spawning and rearing habitat due to anthropogenic changes (e.g., flow modification and channelisation) is known to have dramatic impacts on fish populations. To compensate for habitat losses due to hydropower development, an artificial fluvial habitat channel (‘Compensation Creek’) was constructed in south-central Newfoundland, Canada. The creek was designed to include appropriate habitat features for the two dominant salmonid fish species, landlocked Atlantic salmon (Salmo salar L.) and brook charr (Salvenius fontinalis Mitchell). The study examines the habitat use of landlocked Atlantic salmon and brook charr in the Compensation Creek using electromyogram (EMG) radio telemetry. Ten landlocked Atlantic salmon and eight brook charr were captured and tagged with EMG transmitters. In laboratory swimming experiments, the EMG values were calibrated against swimming speed. Fish were then released in the Compensation Creek and tracked on a daily basis. The results show that (1) during residence in the creek, both species used preferentially the habitat features designed to match their rearing habitat preferences, and (2) swimming speed did not vary among habitat types for either species.     

Oxidative stress and antioxidant defenses in goldfish Carassius auratus during anoxia and reoxygenation

The purpose of this work was to evaluate the response of the antioxidant system of goldfish Carassius auratus during anoxia and reoxygenation. The exposure of goldfish to 8 h of anoxia induced a 14% decrease in total glutathione levels in the kidney, although the liver, brain, and muscle were unaffected. Anoxia also resulted in increases in the activities of liver catalase, brain glucose-6-phosphate dehydrogenase, and brain glutathione peroxidase (by 38, 26, and 79%, respectively) and a decrease in kidney catalase activity (by 17.5%). After 14 h of reoxygenation, liver catalase and brain glutathione peroxidase activities remained higher than controls and several other tissue-specific changes occurred in enzyme activities. Superoxide dismutase activity was unaffected by anoxia and reoxygenation. The levels of conjugated dienes, as indicators of lipid peroxidation, increased by 114% in liver after 1 h of reoxygenation and by 75% in brain after 14 h of reoxygenation. Lipid peroxidation was unaffected in kidney and depressed during anoxia and reoxygenation (by 44-61%) in muscle. Regulation of the goldfish antioxidant system during anoxia may constitute a biochemical mechanism that minimizes oxidative stress following reoxygenation.     

Behavioural plasticity of lake charr (Salvelinus namaycush) x brook charr (S. fontinalis) F1 hybrids in response to varying social environment

Lake x brook F₁ hybrid charr were observed in heterogeneous groups with lake charr or brook charr to assess the effects of different species on hybrid social behaviour. Levels of behavioural plasticity were low in relation to the behavioural differences between the parental and hybrid types. Feeding behaviour showed the most dramatic change, with higher levels in the lake charr treatment and lower levels in the brook charr treatment than controls, suggesting social facilitation. Even though only two measures varied significantly between treatments, there was a tendency for the frequencies and durations of all agonistic measures to increase in the presence of lake charr and decrease with brook charr. A cost-benefit argument is presented to account for this trend.     

Attraction of slimy sculpins to chemical cues of brook charr eggs

In a laboratory study to examine the responses of slimy sculpin Cottus cognatus to chemical cues of brook charr Salvelinus fontinalis eggs, water that held freshly laid eggs (<20 min old), water-hardened eggs (i.e. eggs >4 h old) or injured eggs was collected and used in a series of two-choice tests. Slimy sculpin were exposed to paired stimuli of (1) hard egg water v . control water, (2) fresh egg water v . control water, (3) hard egg water v . injured egg water or (4) hard egg water v . fresh egg water. Sculpin spent considerably more time on the side of the tank with hard egg water and fresh egg water v . control water and injured egg water v . hard egg water. Sculpin did not show a preference for hard egg water v . fresh egg water. In a field study, brook charr were attracted to chemical cues from brook charr eggs, suggesting that brook charr eggs produce sufficient odour to attract some species under natural conditions.     

Regulation of alcoholic fermentation in coleoptiles of two rice cultivars differing in tolerance to anoxia

To investigate regulation of anaerobic carbohydrate catabolism in anoxia-tolerant plant tissue, rate of alcoholic fermentation and maximum catalytic activities of four key enzymes were assessed in coleoptiles of two rice cultivars that differ in tolerance to anoxia. The enzymes were ATP-dependent phosphofructokinase (PFK), pyrophosphate-dependent phosphofructokinase (PFP), pyruvate decarboxylase (PDC), and alcohol dehydrogenase (ADH). During anoxia, rates of coleoptile elongation and ethanol synthesis were faster in the more tolerant variety Calrose than in IR22. Calrose coleoptiles, in contrast to IR22, also showed a sustained Pasteur effect, with the estimated rate of glycolysis during anoxia being 1.4-1.7-fold faster than that of aerobic coleoptiles. In Calrose after 5 d anoxia, maximum catalytic activities of crude enzyme extracts were (in mumol substrate g-1 fresh weight min.-1) 170-240 for ADH, 4-6 for PDC and PFP and 0.4-0.7 for PFK. During anoxia, activity per coleoptile of all four enzymes increased 3-5.5-fold, suggesting that PFK, and PFP, like PDC and ADH, are synthesised in anoxic rice coleoptiles. Enzyme activities, on a fresh weight basis, were lower in IR22 than in Calrose. In vivo activities of PDC and PFK in anoxic coleoptiles from both cultivars were calculated using in vitro activities, estimated substrate levels, cytoplasmic pH, and S0.5 (the substrate level at which 0.5Vmax is reached, without inferring Michaelis-Menten kinetics). Data indicated that potential carbon flux through PFK, rather than through PDC, more closely approximated rates of alcoholic fermentation. That PFK is an important site of regulation was supported further for Calrose coleoptiles by a decrease in the concentration of its substrate pool (F-6-P + G-6-P) following the onset of anoxia. By contrast, in IR22, there was little evidence for control by PFK, consistent with recent evidence that suggests substrate supply limits alcoholic fermentation in this cultivar.     

Phenotypic responses of lacustrine brook charr in relation to the intensity of interspecific competition

We investigated the structure of interindividual variations in the diet of brook charr (Salvelinus fontinalis) based on stomach contents data of 3776 charr captured in 69 lakes of the Canadian Shield (Québec, Canada); 29 of these contained allopatric brook charr populations, 24 contained brook charr and creek chub (Semotilus atromaculatus) and 16 contained brook charr and white sucker (Catostomus commersoni). In any given lake, some of the charr fed almost exclusively on benthic organisms (benthic specialists, i.e., mean percent weight of benthic prey >90%), others, almost exclusively on pelagic prey (pelagic specialists, i.e., mean percent weight of benthic prey <10%), and a lesser proportion were generalist feeders (i.e. mean percent weight of benthic prey between 10 and 90%). The proportion of benthic and pelagic specialists were respectively 41.3 and 18% in allopatric brook charr populations. These proportions fit remarkably well with those based on interindividual preferences in spatial distribution, identified through radio-telemetry in another study done in two lakes of the same area. The proportion of benthic specialists was related to competition for benthic organisms with creek chub and white sucker. The effect of white sucker on brook charr diet was more pronounced than that of creek chub: the proportion of benthic specialists among brook charr decreased from 41.3% in allopatry to 19.7% in sympatry with creek chub, and to 9.9% in sympatry with white sucker. Other response variables of brook charr populations also indicate that white sucker is a stronger competitor than creek chub in this system. Because sucker and chub were introduced in these lakes during the last century, phenotypic responses of brook charr to interspecific competition appear to be rapid. Furthermore, in addition to providing a strong field support to the current hypothesis that polymorphism is promoted by arelaxation of interspecific competition, our results also indicate that phenotypic response of brook charr (i.e. the proportion of each form in a given lake) is related to the intensity of this competition.     

Mitochondrial DNA variation of Salvelinus species found in Finland

Mitochondrial DNA (mtDNA) was purified from the Arctic charr, Salvelinus alpinus , the brook charr, Salvelinus fontinalis , and the lake charr, Salvelinus namaycush , and digested with restriction enzymes Ava II, Hinf I, Eco R V, Pst I and Xba I. Two Arctic charr samples were from natural populations and they represented two different morphotypes of Arctic charr. All other studied populations were hatchery maintained. Eight additional restriction enzymes and double digestions were employed to study morphotypes of Arctic charr. We distinguished two morphotypes with restriction enzyme Nci I. Sequence divergence among mtDNA types was 2.9–3.8% between S. alpinus and S. fontinalis , 3.4–4.6% between S. alpinus and S. namaycush , and 4.7–5.3% between S. fontinalis and S. namaycush . lntraspecific variation was lowest in Arctic charr, the average of nucleon diversity for three populations being 0.179, while for brook charr and for lake charr nucleon diversity was 0.334 and 0.550, respectively. According to the number of mtDNA types, it is obvious that introduction to Finland and hatchery propagation have not greatly affected the mtDNA variation of brook charr or lake charr.     

Comparative activities of glycogen phosphorylase and gamma-amylase in livers of carp (Cyprinus carpio) and goldfish (Carassius auratus).

1. The activity of glycogen phosphorylase in goldfish liver is fivefold greater than that in carp liver, suggesting that the enzyme may not be as important in regulating glycogenolysis in the latter species. 2. The activity of gamma-amylase is comparable in carp and goldfish liver. 3. The activity of hepatic gamma-amylase is approximately one-half that of glycogen phosphorylase in carp whereas in goldfish, the activity of gamma-amylase is less than one-sixth that of phosphorylase. Hepatic gamma-amylase may be an important glycogenolytic enzyme in carp but makes an insignificant contribution to glycogenolysis in goldfish.