Effect of β-adrenergic stimulation of trout erythrocytes on blood viscosity

1. Both the viscosity of trout blood and its dependence on shear rate are reduced when erythrocyte β-adrenoceptors are stimulated.2. This suggests that catecholamines released during stress may decrease the resistance of nucleated erythrocytes to blood flow.     

Microsatellite DNA analysis of rainbow trout (Oncorhynchus mykiss) from western Alberta, Canada: native status and evolutionary distinctiveness of “Athabasca” rainbow trout

Molecular genetic assays can contribute to conservation of aquatic taxa by assessing evolutionary and taxonomic distinctiveness, levels of genetic variation within and between populations, and the degree of introgression with introduced taxa. The Athabasca River drainage of␣western Alberta, Canada is one of only three (and the largest) drainages flowing east of the continental divide that contain native populations of rainbow trout (Salmonidae: Oncorhynchus mykiss). The “Athabasca” rainbow trout has been considered a preglacial relict worthy of special conservation measures. In addition, the native range of Athabasca rainbow trout has seen many instances of introductions of non-native populations since the beginning of the 20th century. We assayed rainbow trout from the Athabasca River drainage, from hatchery populations, and from representative populations in adjacent regions (N = 49 localities) for variation at 10 microsatelite loci to assess the level of evolutionary distinctiveness of Athabasca rainbow trout, and to assess the levels of introgression with non-native hatchery fish. We found that native Athabasca rainbow trout did not form a distinctive genetic assemblage and that the greatest amount of allele frequency variation was attributable to contemporary drainage systems (29.3%) rather than by a Athabasca/non-Athabasca distinction (12.6%). We found that 78% of all fish were confidently assigned to a “wild” rather than a “hatchery” genetic grouping and that most of the inferred introgression with hatchery fish was restricted to a few localities (N = 6). Our results suggest that: (i)␣Athabasca River rainbow trout are likely postglacial immigrants from adjacent populations of the Fraser River, and (ii) that there is no evidence of widespread introgression of hatchery alleles into native Athabasca River drainage rainbow trout.     

Does socially induced stress in rainbow trout cause chloride cell proliferation?

At the end of a 2-week confinement period, subordinate rainbow trout Oncorhynchus mykiss had significantly higher circulating concentrations of plasma cortisol than did the dominant fish with which they were paired. Physiological effects linked to elevated plasma cortisol concentrations in subordinate fish included loss of weight and a lowering of condition factor. However, there were no significant differences in gill epithelium chloride cell numbers or blood plasma ion concentrations between dominant and subordinate fish. It is concluded that elevated plasma cortisol concentrations elicited by the social stressors of the present study did not cause proliferation of chloride cells.     

Does prolactin affect steroid secretion by isolated rainbow trout ovarian cells?

The in vitro secretion of progesterone (P(4)), androgen (A) and estradiol (E(2)) by follicular cells, isolated monthly from the rainbow trout ovaries during the whole annual cycle, was studied. Cells were cultured as monolayers in control and prolactin (PRL) supplemented media. E(2) secretion showed two distinct maxima in September and January: 4959+/-220 pg/ml and 3166+/-121 pg/ml, respectively, i.e. during vitellogenesis and before the spawning time. PRL had a significant (16%) suppressive effect on E(2) secretion when the level of secreted steroid was at its highest (4167+/-193 pg/ml) at the end of vitellogenesis and by 32% (2157+/-124 pg/ml), before ovulation. Increased P(4) levels observed in February (988+/-69 pg/ml) and March (2008+/-74 pg/ml) may be connected with the need for a substrate for the synthesis of 17alpha20betaOH-P (MIS). At this time, the secretion of P(4) was also suppressed by PRL and was reduced to 1395+/-78 pg/ml. Our results indicate that PRL may play a role in fish reproduction.     

Evidence for NO-dependent vasodilation in the trout (Oncorhynchus mykiss ) coronary system

The effects of l-arginine, and its analogues N ^ω-nitro-l-arginine methyl ester and N ^ω-nitro-l-arginine on vascular resistance were investigated in the intact coronary system of an isolated non-working trout heart preparation. l-Arginine, at 10^–8 mol · l^–1induced a slight vasodilatory effect (max 10%). N ^ω-nitro-l-arginine methyl ester and N ^ω-Nitro-l-arginine in the range 10^–8–10^–4 mol · l^–1 caused dose-dependent increases in coronary resistance. The vasodilatory action of l-arginine was abolished when the preparation was pretreated with 10^–4 mol · l^–1 N ^ω-nitro-l-arginine or N ^ω-nitro-l-arginine methyl ester. Nitroprusside alone at 1 mmol · l^–1 induced a maximum vasodilation (30%) of the coronary system. Methylene blue a known inhibitor of guanylate cyclase, induced a strong vasoconstriction (already significant at 10^–5 mol · l^–1) and was able to overcome the vasodilative effect of nitroprusside. The endothelial nitric oxide agonists acetylcholine and serotonin, established in mammalian vessels, also mediate vasodilation in trout coronary system. In 50% of preparations, acetylcholine induced a biphasic response with vasodilation at low concentration (max 15% at 10^–8 mol · l^–1). Serotonin displayed a dose-response vasodilation in the range 10^–8–10^–4 mol · l^–1 (max 20%). These vasodilative effects were reduced or abolished by 10^–4 mol · l^–1 l-NA. These data support the existence of NO-mediated vasodilation mechanisms in the trout coronary system. Accepted: 1 July 1996     

Allosteric modulation by S-nitrosation in the low-O₂ affinity myoglobin from rainbow trout

Myoglobin (Mb) serves in the facilitated diffusion and storage of O? in heart and skeletal muscle, where it also regulates O? consumption via nitric oxide (NO) scavenging or generation. S-nitrosation at reactive cysteines may generate S-nitroso Mb (Mb-SNO) and contribute further to NO homeostasis. In being a monomer, Mb is commonly believed to lack allosteric control of heme reactivity. Here, we test whether in rainbow trout, a fast swimmer living in well-aerated water, the Mb-O? affinity is regulated by ionic cofactors and S-nitrosation. O? equilibria showed the lowest O? affinity ever reported among vertebrate Mbs (P?? = 4.92 ± 0.29 mmHg, 25°C), a small overall heat of oxygenation (ΔH = -12.03 kcal/mol O?), and no effect of chloride, pH, or lactate. Although the reaction with 4,4'-dithiodipyridine (4-PDS) showed 1.3-1.9 accessible thiols per heme, the reaction of Mb with S-nitroso cysteine (Cys-NO) and S-nitrosoglutathione (GSNO) to generate Mb-SNO yielded ～0.3-0.6 and ～0.1 SNO/heme, respectively, suggesting S-nitrosation at only one cysteine (likely Cys1?). At ～60% S-nitrosation, trout Mb-SNO showed a higher O? affinity (P?? = 2.23 ± 0.19 mmHg, 20°C) than unmodified Mb (3.36 ± 0.11 mmHg, 20°C). Total SNO levels measured by chemiluminescence in trout myocardial preparations decreased after hypoxia, but not significantly, indicating that transnitrosation reactions between thiols may occur in vivo. Our data reveal a novel, S-nitrosation-dependent allosteric mechanism in this low-affinity Mb that may contribute to targeted O?-linked SNO release in the hypoxic fish heart and be of importance in preserving cardiac function during intense exercise.     

Restoring marble trout genes in the Soča River (Slovenia)

In Slovenia, the unique watershed naturally hosting the marble trout is the So?a River, called Isonzo in Italy. In 1993–1996 molecular data established the existence of extensive hybridization with stocked Atlantic domestic lineages which is a threat for this taxon and for the economy of the country established on the angling tourism. Different management actions have been developed for restoring marble genes since 1996: banning stocking of brown trout, revising fishing regulations for anglers and testing genetically brood stock in hatchery for stocking phenotypic and pure marble fry. This long fight against hybridization was genetically surveyed using allozymes, mitochondrial sequences and microsatellites according to the available technique at each period. Despite the irregularity of genotyping along nearly fifteen years after the new management started, it appears that the proportion of domestic lineage in the river dropped regularly of about 2% each year, a positive result for conservative management measures.

     

Zinc-dependent binding between peptides derived from rainbow trout CD8α and LCK

The cytoplasmic tail of mammalian CD8α binds the kinase LCK in a zinc-dependent manner. In analogy with a previous study for humans (Kim et al., 2003) peptides were synthesized from rainbow trout CD8α and LCK. Surface plasmon resonance (SPR) analysis indicated that also in fish these molecules bind to each other in a zinc-dependent manner.     

Inhibition of in vitro aflatoxin B1-DNA binding in rainbow trout by CYP1A inhibitors: α-naphthoflavone, β-naphthoflavone and trout CYP1A1 peptide antibody

Rainbow trout cytochrome P450 (CYP)1A detoxifies aflatoxin B₁ (AFB₁) to aflatoxin M₁ (AFM₁), whereas CYP2K1 activates AFB₁ to AFB₁-8,9-epoxide. We report that α-naphthoflavone (ANF) and β-naphthoflavone (BNF) both strongly inhibit CYP1A-mediated ethoxyresorufin O-deethylase (EROD) activity (K_i = 9.1 ± 0.8 and 7.6 ± 1.1 nM, respectively). These inhibitors (selective for mammalian CYP1A at low concentrations), as well as rabbit polyclonal antibody to a trout CYP1A1 peptide (residues 277–294), also strongly inhibited trout microsome-catalyzed AFB₁-DNA binding and lauric acid (ω-1) hydroxylation in vitro, reactions previously established to be CYP2K1-dependent. ANF at 0.5, 5, 50 and 500 μM inhibited liver microsome-catalyzed AFB₁-DNA binding by 22, 58, 84 and 91%, respectively, whereas BNF at the same concentrations inhibited 22, 74, 78 and 81%, respectively. The CYP1A1 peptide and CYP2K1 polyclonal antibodies (10 mg IgG/mg microsomal protein) inhibited AFB₁-DNA binding by 84 and 66%, respectively, compared with pre-immune IgG. Lauric acid (ω-1) hydroxylation was inhibited 61% by 5 μM ANF, 69% by 5 μM BNF and 100% by either antibody at 12 mg IgG/mg microsomal protein. These results demonstrate that mammalian CYP1A inhibitors also inhibit trout microsomal AFB₁-DNA binding and lauric acid (ω-1) hydroxylation, catalyzed primarily by CYP2K1. In the absence of evidence that trout CYP1A can catalyze AFB₁-DNA binding, the results suggest configuration similarities at, or near, the active sites for these two fish enzymes that result in antibody crossreaction and loss of the inhibitor specificity observed with mammalian CYP1A.     

Does bradycardia or hypertension enhance gas transfer in rainbow trout (Oncorhynchus mykiss)?

Experiments were conducted to test the hypothesis that branchial gas transfer is enhanced in rainbow trout during hypoxia or hypercarbia by bradycardia and systemic vasoconstriction. Gas transfer was indirectly assessed by continuous monitoring of arterial blood gases, PaO2 and PaCO2. Cardiac frequency was maximally decreased by 34.9+/-4.3 and 8.6+/-3.2 bpm in hypoxic and hypercarbic fish, respectively. Pre-treating fish with atropine (1micromol kg(-1)) attenuated or abolished the bradycardia during hypoxia and hypercarbia, respectively. However, there were no significant differences in the arterial blood gases between the control and atropinized fish. Dorsal aortic blood pressure was increased maximally by 11.3+/-2.8 and 17.7+/-2.0mm Hg in the hypoxic and hypercarbic fish. Pre-treatment of fish with prazosin (2.4micromol kg(-1)) prevented these increases in blood pressure. Blood gases were unaltered by prazosin treatment in the hypercarbic fish. However, in the hypoxic fish, gas transfer appeared to be impaired by prazosin on the basis of lowered PaO2 (by approximately 35 mm Hg compared to control fish) and increased PaCO2 (by approximately 0.3mm Hg). Because the normal hyperventilatory response to hypoxia was prevented by prazosin, it is possible that the impairment of gas transfer was related to inadequate ventilation rather than to any differences in the pressor response. The present results provide no evidence that gas transfer in rainbow trout is enhanced by bradycardia nor do they reveal any obvious benefit associated with the increases in blood pressure that accompany hypoxia and hypercarbia.     

How repeatable is CTmax within individual brook trout over short- and long-time intervals?

As stream temperatures increase due to factors such as heated runoff from impervious surfaces, deforestation, and climate change, fish species adapted to cold water streams are forced to move to more suitable habitat, acclimate or adapt to increased thermal regimes, or die. To estimate the potential for adaptation, a (within individual) repeatable metric of thermal tolerance is imperative. Critical thermal maximum (CT_max) is a dynamic test that is widely used to measure thermal tolerance across many taxa and has been used in fishes for decades, but its repeatability in most species is unknown. CT_max tests increase water temperature steadily over time until loss of equilibrium (LOE) is achieved. To determine if CT_max is a consistent metric within individual fish, we measured CT_max on the same lab-held individually-marked adult brook trout Salvelinus fontinalis at three different times (August & September 2016, September 2017). We found that CT_max is a repeatable trait (Repeatability ± S.E.: 0.48 ± 0.14). CT_max of individuals males was consistent over time, but the CT_max of females increased slightly over time. This result indicates that CT_max is a robust, repeatable estimate of thermal tolerance in a cold-water adapted fish.     

Non-indigenous brook trout and the demise of Pacific salmon: a forgotten threat?

Non-indigenous species may be the most severe environmental threat the world now faces. Fishes, in particular, have been intentionally introduced worldwide and have commonly caused the local extinction of native fish. Despite their importance, the impact of introduced fishes on threatened populations of Pacific salmon has never been systemically examined. Here, we take advantage of several unique datasets from the Columbia River Basin to address the impact of non-indigenous brook trout, Salvelinus fontinalis, on threatened spring/summer-run chinook salmon, Oncorhynchus tshawytscha. More than 41 000 juvenile chinook were individually marked, and their survival in streams without brook trout was nearly double the survival in streams with brook trout. Furthermore, when brook trout were absent, habitat quality was positively associated with chinook survival, but when brook trout were present no relationship between chinook survival and habitat quality was evident. The difference in juvenile chinook survival between sites with, and without, brook trout would increase population growth rate (lambda) by ca. 2.5%. This increase in lambda would be sufficient to reverse the negative population growth observed in many chinook populations. Because many of the populations we investigated occur in wilderness areas, their habitat has been considered pristine; however, our results emphasize that non-indigenous species are present and may have a dramatic impact, even in remote regions that otherwise appear pristine.     

Compensatory growth for free? A field experiment on brown trout,Salmo trutta

Laboratory studies suggest that animals may be capable of compensatory growth after periods of food shortage. There is, however, a lack of field experiments investigating the incidence and consequences of compensatory growth in the wild, and the relevance of compensatory responses in natural populations has recently been questioned. Here we addressed the hypotheses that (1) food restriction during critical growth periods can induce compensatory growth, and (2) that compensatory growth is associated with delayed costs in natural populations. These hypotheses were addressed by (1) manipulating the food intake of brown trout in spring, (2) measuring growth rate responses over the first month following release, and (3) measuring growth and mortality (i.e. recapture rate) over the subsequent fall and winter. We found that brown trout restored lost body weight and condition within a month, providing the first experimental demonstration of compensatory growth in the wild. However, no delayed costs of the compensatory response could be detected within the timespan of the experiment. We suggest that wild brown trout have an adapted "buffer capacity" to withstand fluctuations in food supply, allowing restoration of lost lipid reserves when feeding conditions improve. However, when prolonged food deprivation affect structural components, compensation may not be possible without compromising long-term performance.     

Temperature dependence of cardiac sarcoplasmic reticulum Ca²⁺-ATPase from rainbow trout Oncorhynchus mykiss

In this work, the temperature dependence of the sarco-endoplasmic reticulum Ca(2+) -ATPase (SERCA2) activity from rainbow trout Oncorhynchus mykiss cardiac ventricles was measured and compared with the mammalian SERCA2 isoform. The rate of ATP-dependent Ca(2+) transport catalysed by O. mykiss vesicles was totally abolished by thapsigargin and the Ca(2+) ionophore A(23187) . At warm temperatures (25 and 30° C), the SERCA2 from O. mykiss ventricles displayed the same rate of Ca(2+) uptake. At 35° C, the activity of the O. mykiss enzyme decreased after 20 min of reaction time. The rate of Ca(2+) uptake catalysed by the mammalian SERCA2 was temperature dependent exhibiting its maximal activity at 35° C. In contrast to the rate of Ca(2+) uptake, the rate of ATP hydrolysis catalysed by O. mykiss SERCA2 was not significantly different at 25 and 35° C, but the rate of ATP hydrolysis catalysed by the rat Rattus norvegicus SERCA2 isoform at 35° C was two-fold higher than at 25° C. At low temperatures (5 to 20° C), the rate of Ca(2+) uptake from O. mykiss SR was less temperature dependent than the R. norvegicus isoform, being able to sustain a high activity even at 5° C. The mean ±s.e. Q(10) values calculated from 25 to 35° C for ATP hydrolysis were 1·112 ± 0·026 (n = 3) and 2·759 ± 0·240 (n = 5) for O. mykiss and R. norvegicus, respectively. Taken together, the results show that the O. mykiss SERCA2 was not temperature dependent over the 10 to 25° C temperature interval commonly experienced by the animal in vivo. The Q(10) value of SERCA2 was significantly lower in O. mykiss than R. norvegicus which may be key for cardiac function over the wide environmental temperatures experienced in this eurythermal fish.     

Kin-biased distribution in brown trout: an effect of redd location or kin recognition?

A wide range of animals have been reported to show kin-biased behaviours, such as reduced aggressiveness and increased food sharing among relatives. However, less is known about whether wild animals also associate with relatives under natural conditions, which is a prerequisite to facilitate kin-biased behaviours and hence kin selection. We tested, by means of microsatellite polymorphism, correlations between pair-wise relatedness and pair-wise metric distance in wild brown trout (Salmo trutta L.) under natural conditions in two streams. Our data show that young-of-the-year as well as older trout found close together also had a higher genetic relatedness in one of the two streams, whereas no relationship was found in the other stream. Very few half and full siblings were found in the second stream and under these conditions it is unlikely that kin-biased behaviours will receive positive selection. We discuss the underlying mechanisms for the observed structure and we specifically address the issue of whether the grouping of related individuals could reflect dispersal from the same spawning redds, or if it reflects active association with relatives, possibly conferring kin-selected advantages.     

Interspecific hybridization, a matter of pioneering? Insights from Atlantic salmon and brown trout

Interspecific hybridization may occur in situations of recent contact between a colonizer and a resident species, being more intense in the colonization front. Atlantic salmon Salmo salar and brown trout S. trutta have been sympatric species since their origin and they share spatial and temporal spawning niches, exhibiting low levels of bidirectional interspecific hybridization and introgression throughout their distribution range. Different causes have been identified for increased hybridization, from escapes or deliberate releases of domesticated fish to sneaking male behavior. We have examined hybridization rates and direction in different situations of advance of one of these species into a territory formerly inhabited by the other (247 samples were analyzed in northern Spain and 487 in Kerguelen Islands). In all cases, hybrids found in the colonization front were offspring of colonizer females and resident males. We hypothesize that these findings are the result of adaptive relaxed mate choice of colonizing females, regardless of the relative abundance of each species.     

Invasive brown trout Salmo trutta induce differential growth strategies in the native snow trout Schizothorax richardsonii of Himalaya: Are natives in unaltered rivers better at picking the gauntlet of invasion?

Brown trout Salmo trutta is a potent global invader and its establishments have progressively altered physiologies, life-histories and niche-availabilities for native fish species. River impoundments further escalate its invasion potential. The Himalayan rivers however, stay uncharted for the effects of brown trout interactions with the native fish fauna. Snow trout Schizothorax richardsonii a Himalayan cold-water native, concerningly overlaps its range with brown trout. To understand its responses to invasion pressures, we investigated brown trout effects on the age and growth of snow trout populations in three rivers with varying levels of perturbation: (a) a dammed and (b) an undammed river with the invasive brown trout in comparison to (c) an undammed river without invasion pressures. We found sympatric snow trout in the undammed river to respond to brown trout invasion with fast life history responses, showing an early age-at-maturity (A₅₀ = 1.2 years) and fast growth with a higher growth constant (K = 0.40 yr⁻¹) and specific rate of linear growth across life. On the contrary, sympatric snow trout in the dammed river showed an explicitly slow life-history by maturing at a higher age (A₅₀ = 2.9 years) and a slow growth, with a lower growth constant (K = 0.26 yr⁻¹) and specific linear growth rates. Our findings suggest that, the snow trout appear to present stronger response to brown trout invasions when the river is unaltered and free from hydropower operations and damming. Further research is strongly warranted from other high-altitude Himalayan basins to delineate the variation in growth strategies exhibited by snow trout in sympatry with the invasives.