Blood parameters in brook trout Salvelinus fontinalis (Mitchill, 1815), affected by columnaris disease

Mass death of farmed brook trout Salvelinus fontinalis reared in raceway culture on a commercial fish farm was recorded in May. The condition was caused by natural infection with external signs of Flavobacterium columnare. Haematological analyses were carried out from peripheral blood from 10 fish from each group. Anaemia was characterized by a reduced erythrocyte count and lower haemoglobin levels; the values of mean corpuscular volume and mean corpuscular haemoglobin were higher and the values of mean corpuscular haemoglobin concentration were decreased. Clinical chemistry analyses in blood plasma indicated decreased levels of total protein, glucose and total calcium and an increase in the urea level. Among the four enzymes analysed, a catalytic activity reaching multiples of the normal level was found in alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase. The catalytic concentration of alkaline phosphatase was decreased. Electrophoretic analyses of the plasma protein indicated a reduced level of albumin. The moribund fish had a higher hepatosomatic index and their condition coefficient after Fulton and after Clark was also higher.     

Angiotensins stimulate in vitro ovulation and contraction of brook trout (Salvelinus fontinalis) follicles

The effects of salmon angiotensin I (sAI) and human angiotensin II (hAII) on in vitro ovulation of preovulatory (preOV) brook trout (Salvelinus fontinalis) follicles were investigated. Both angiotensins increased levels of ovulation above that in controls after 12 hours of incubation. The increase was statistically significant in incubates with greater than 1 M hAII. The effects of the angiotensins on follicle contraction were also studied indirectly by measuring the decrease in weight of punctured follicles taken prior to germinal vesicle breakdown. At 1 M, both angiotensins significantly decreased the weight of punctured follicles after 16 hours of incubation. The angiotensin-stimulated decrease in weight was not blocked by indomethacin (10 g/ml), indicating that follicle contraction was not prostaglandin-dependent. The data indicate that angiotensins might be directly involved in brook trout ovulation and the stimulatory effects of angiotensins on ovulation may be attributed to their effects on follicle contraction.This work was supported by NIH grant #HD25924-02.     

Volume regulation in glutathione-treated brook trout (Salvelinus fontinalis) erythrocytes

Brook trout erythrocytes that were washed with and suspended in Ringer's solution with reduced glutathione (1.0 mM) maintained steady state cell volume for up to 24h, while those without the thiol-protective agent steadily shrank. Changes in cell volume (measured as packed cell volume, PCV) were evoked by acidic media (Ringer's at pH 6.8), hypoosmotic solutions (or both) and intracellular K⁺ and Cl⁻ concentrations were monitored over 4h. Acid-swollen cells failed to volume regulate or release K⁺ but had significantly elevated intracellular Cl⁻ Osmotically-swollen cells at pH 7.8 but not at pH 6.8 underwent regulatory volume decrease (RVD) and returned to initial levels in 2h, accompanied by release of K⁺ and Cl⁻ In contrast, osmotically-shrunken cells did not show regulatory volume increase. The regulatory volume decrease and concomitant K⁺ release were dependent on Cl⁻ implying a direct or indirect coupling of K⁺ to Cl⁻ transport in volume regulation. RVD was partially blocked by 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS, 0.1 mM), an anion exchange blocker, but was unaffected by amiloride (1.0 mM) which blocks Na⁺/H⁺ exchange. Amiloride and DIDS prevented the swelling response to low pH but had no effect on control cells, suggesting involvement of Na⁺/H⁺ and Cl⁻/HCO₃ ⁻ exchanges in acid-induced cell swelling. Quinine (1.0 mM) a known blocker of K⁺ channels, exacerbated the osmotically-induced swelling but had little effect on the subsequent RVD and release of KCl. The results suggest that low extracellular pH inhibits neutral C⁻-dependent K⁺ release and the resultant regulatory volume decrease in osmotically-swollen cells.     

Growth variability of brook charr (Salvelinus fontinalis) in coastal maine

Brook charr inhabit a wide variety of habitats and exhibit considerable growth variation within and among populations. We used biannual mark–recapture electrofishing surveys and PIT tags at two streams in coastal Maine to examine spatial and temporal variability in brook charr growth. We documented considerable stream‐to‐stream and year‐to‐year variation in the growth of coastal brook charr. Based on 2892 recaptures between 2006 and 2010, we developed a suite of linear mixed models to examine variation in individual growth rates at Stanley Brook as a function of environmental variables. We distilled physical habitat data from surveys in 2010 into principle components for entry into the growth model. Growth was related primarily to an interaction between season and body length, mean water temperature, and instream location. Growth rates were highest in the summer, and smaller individuals grew more rapidly than larger conspecifics in the same stream. Individuals using habitats closer to the head of tide grew faster than those in upstream locations. Physical habitat variation was not an important control of individual growth variation within Stanley Brook. We suggest local variability in growth rates should be considered when developing management strategies for coastal populations.     

Cortisol,glucose and hematocrit changes during acute stress,cohort sampling,and the diel cycle in diploid and triploid brook trout (Salvelinus fontinalis Mitchill)

In this study, groups of diploid (control) and triploid brook trout were subjected to a 5 min acute handling/confinement stress. Blood samples were collected from equal numbers of diploid and triploid fish at regular predetermined intervals, beginning prior to confinement and continuing for up to 3h after confinement. These blood samples were used for the measurement of hematocrit and plasma cortisol and glucose concentrations. The effects of cohort sampling and the diel cycle on these variables were also analysed. Except for minor differences, both groups responded similarly to the acute stress, with short-term elevations in plasma cortisol and hematocrit levels, and a more sustained increase in plasma glucose levels. No significant differences were found between diploid and triploid fish in their reactions to cohort sampling. However, the act of removing an individual from a tank caused significant changes in plasma cortisol levels in cohorts remaining in that tank (p < 0.001). Significant diel effects were observed for both ploidy groups in plasma cortisol (p < 0.05) but not in plasma glucose or hematocrit. To whom correspondence should be addressed     

Seasonal habitat use by brook trout,Salvelinus fontinalis (Mitchill), in a second-order stream

Abstract Seasonal habitat use by over-yearling and under-yearling brook trout, Salvelinus fontinalis (Mitchill), was examined in a second-order stream in north-central Pennsylvania, USA. The habitat occupied by brook trout and available habitat were determined in a 0.5-km stream reach during the spring, summer and autumn of 1989 and the spring and summer of 1990. Cover, depth, substrate and velocity were quantified from over 2000 observations of individual brook trout. Habitat used by under-yearling brook trout was more uniform between seasons and years than that used by over-yearling brook trout. Over-yearling brook trout occupied areas with more cover and greater depth than did under-yearling brook trout, suggesting ontogenetic shifts in these variables. Differences for velocity and substrate were not as great as those for cover and depth. The selection of areas with low water velocities governed trout habitat use in spring, whereas cover and depth were the most important habitat variables in summer and autumn. Principal component analysis showed that available habitat and trout habitat centroids diverged most in spring, indicating that habitat selection by brook trout may be greatest at this time.     

Protein and energy nutrition of brook trout (Salvelinus fontinalis) at optimal and elevated temperatures

The digestible protein (DP) and digestible energy (DE) requirements for maintenance and growth of brook trout (Salvelinus fontinalis) were determined using a factorial model at either optimum (15 °C) or elevated temperature (19 °C). Several key parameters of the factorial model were measured using a series of inter‐related studies. The maintenance requirements for DP and DE were 0.10 gDP kg^?0.69 day^?1 (15 °C) and 0.31 gDP kg^?0.78 day^?1 (19 °C), and 34.86 kJDE kg^?0.84 day^?1 (15 °C) and 46.14 kJDE kg^?0.86 day^?1 (19 °C). The total requirements for DP were 0.10 gDP kg^?0.69 day^?1 + 2.14PG (protein gain) (15 °C) and 0.31 gDP kg^?0.78 day^?1 + 1.98PG (19 °C). The total requirements for DE were 36.86 kJDE kg^?0.84 day^?1 + 1.58EG (energy gain) (15 °C) and 46.14 kJDE kg^?0.86 day^?1 + 1.64EG (19 °C). The partial efficiencies for growth were 0.47 (15 °C) and 0.51 (19 °C) for protein, and 0.63 (15 °C) and 0.61 (19 °C) for energy. Nutrient gain was lower at the elevated temperature; however, feed formulation for brook trout should be adjusted to match changes in nutrient requirements at different culture temperatures. The protein and energy requirements model will be useful for developing commercial feeds and feeding charts for brook trout.     

Expert opinion on the status and stressors of brook trout,Salvelinus fontinalis,in Ontario

Ontario supports a vast fisheries resource with an abundance of lakes, rivers and streams. A landscape approach to management informed by a broad‐scale monitoring programme has been initiated to assess the status of fisheries within lakes. However, not all species are assessed by this programme, and there is no provincial monitoring of species inhabiting rivers and streams. As such, changes in the status of a species such as brook trout, Salvelinus fontinalis (Mitchill), could be entirely missed. Brook trout is a highly valued and sought after species by anglers within the province, but there are concerns the species is declining. Given the paucity of broad, empirical data, the status and trends of brook trout across the province have been based on expert opinion at multiple local scales. In 2016, a online questionnaire was sent to brook trout experts to determine status, stressors, management approaches and assess risks (magnitude and probability) to lake and river/stream populations in different geographic areas of Ontario. A Bayesian network was used to analyse responses and develop a risk assessment based on expert opinion for brook trout at multiple scales within the province.     

Effects of dietary carbohydrate level on growth performance,innate immunity,antioxidant ability and hypoxia resistant of brook trout Salvelinus fontinalis

Juvenile brook trout were fed on six isonitrogenous and isolipidic diets, containing graded levels of gelatinized corn starch (50, 100, 150, 200, 250, 300 g/kg diet) for 81 days. Cellulose was used to compensate carbohydrate loss. The weight gain, specific growth rate and protein efficiency ratio increased as the dietary carbohydrate level increased from 50 to 150 g/kg, but decreased thereafter (p < .05). Quadratic regression analysis revealed that the optimum dietary carbohydrate level was in the range of 187.1–194.1 g/kg. In addition, serum alanine transaminase and aspartate transaminase activities, as well as glucose content, were highest in the group fed on 300 g/kg carbohydrate (p < .05). The hepatic malondialdehyde level increased with dietary carbohydrate levels (p < .05). The hepatic lysozyme activity increased as dietary carbohydrate level increased from 50 to 150 g/kg and decreased thereafter (p < .05). The overall survival rate after hypoxia challenge (45 min; dissolved oxygen content: 2.2 mg L^?1) decreased with dietary carbohydrate levels (p < .05). Taken together, these results suggested that optimal level of dietary carbohydrate could improve growth performance. However, excessive dietary carbohydrate intake (> 250 g/kg) may decrease innate immunity status, increase oxidative stress and reduce resistance to hypoxia stress in brook trout.     

Tissue concentrations of RNA and protein for juvenile brook trout (Salvelinus fontinalis): lagged responses to fluctuations in food availability

To be useful as short-term indices of nutritional status when food availability varies, wet weight-at-length and tissue concentrations of nucleic acids and protein must covary closely with a fish's recent feeding history. We measured changes in these indices for young brook trout (Salvelinus fontinalis) (fork length: 20–34 mm) reared under alternating, 4 to 5 d periods of food provisioning and food deprivation. Weight-at-length corresponded closely with current feeding conditions, being higher when the trout were fed than when they were deprived. Concentrations of RNA and protein, however, did not correspond closely with current feeding conditions. Instead, there were significant carry-over effects whereby responses to feeding conditions experienced in one treatment period were not exhibited until the following treatment period. Food provisioning had positive carry-over effects on concentrations of RNA and protein while food deprivation had negative carry-over effects. Consequently, food-deprived trout sometimes had higher concentrations of RNA and protein than well fed trout. Since wild fish may experience short-term fluctuations in food availability, lagged responses in concentrations of nucleic acids or protein like those reported here could hamper attempts to use these biochemical measures to assess the nutritional status of juvenile fish in the field.Author to whom all correspondence should be addressed.     

Brook trout (Salvelinus fontinalis) population dynamics and mottled sculpin (Cottus bairdi) occurrence in relation to acidic episodes in streams

We monitored water chemistry and populations of brook trout (Salvelinus fontinalis) and mottled sculpins (Cottus bairdi) in 5 streams and some tributaries to determine how the severity of acidic episodes (low pH and high Al) influenced density and distribution of these species. Among streams, Linn Run had the lowest pH (4.8) and highest concentrations of total dissolved Al (>400 μg-l^?1) during periods of high discharge. Densities of brook trout and mottled sculpins were low in Linn Run, but high in tributaries, where pH (6.5) and Al (30 μg-l^?1) remained well below lethal levels. Among the other streams, brook trout density was highest where the acidic episodes were least severe and the density of age-0 brook trout was lowest when precipitation during the early months of the year was above normal. Immigration of brook trout from downstream areas maintained populations in the stream segments subjected to severe episodes. Mottled sculpins were not found in streams with severe episodes.     

The reproductive physiology of three age classes of adult female diploid and triploid brook trout (Salvelinus fontinalis)

Triploid female fish show impaired gametogenesis and are unable to produce viable offspring. The reproductive physiology of artificially-induced triploid female salmonids has been well described up until the time of first sexual maturation in diploids, but few reports exist for older triploids. This study reports the influence of triploidy on growth, ovarian development and reproductive endocrinology among three age classes of female brook trout (Salvelinus fontinalis) in comparison to sibling diploids. Triploids were larger than diploids for most of the study period, but the difference was statistically significant only during maturation and spawning of 2⁺ diploids. Plasma estradiol-17 (E₂), testosterone (T) and vitellogenin (VTG) levels in triploids were generally lower than in diploids, and VTG was the only parameter to show seasonal fluctuations resembling those of diploids. Triploids showed significantly lower GSI and total oocyte number than diploids of similar age, and only half of all triploids sacrificed during the study (n=56) had developing oocytes in their ovaries. At age 3⁺, 13 of 19 triploid females had oocytes at various stages of development, including perinucleolar, yolk vesicle and yolk globule stages. In addition, three of these fish had collectively produced 72 mature stage oocytes. Thus, whereas diploid brook trout can produce mature oocytes as two-year-olds, triploids cannot do so until four years of age, with the number of mature oocytes being greatly reduced.     

The effect of dietary energy level on growth and nutrient utilization by juvenile diploid and triploid brook charr,Salvelinus fontinalis

Triploids are used in aquaculture because they are sterile and do not undergo preharvest loss in flesh quality. Despite this advantage, they do not always perform as well as diploids when fed commercial diets. This study investigated whether differences in dietary energy utilization might explain this reduced performance. Dietary lipid levels were adjusted to supply diets with 22.7, 23.6 or 24.4 MJ kg^?1 gross energy and fed to juvenile diploid and triploid brook charr. Fish were fed to satiation twice daily in a 70‐day growth trial, with samples collected at days 0 and 70 for proximate composition and plasma insulin‐like growth factor I (IGF‐I) analysis. This was followed by a digestibility trial using the same diets to determine apparent digestibility coefficients. Triploids had lower growth rates and condition factor than diploids on all three diets, but there was no effect of ploidy on feed conversion efficiency. Triploids also had lower whole‐body lipid content, but equal protein content, which resulted in lower energy content at both days 0 and 70. There was no effect of diet or ploidy on plasma IGF‐I concentrations. Triploidy did not affect diet digestibility, suggesting that the reduced performance of triploids is due to a difference in metabolism and energy utilization.     

Brook trout (Salvelinus fontinalis) movement and survival after removal of two dams on the West Branch of the Wolf River,Wisconsin

Dam removals allow fish to access habitats that may provide ecological benefits and risks, but the extent of fish movements through former dam sites has not been thoroughly evaluated for many species. We installed stationary PIT antennas in 2016 and 2017 to evaluate movements and survival of brook trout Salvelinus fontinalis in the West Branch of the Wolf River (WBWR) in central Wisconsin following removal of two dams and channel modifications designed to promote fish movement. These changes provided access to lacustrine habitats that might provide suitable winter habitat or act as ecological sinks. We used multistate models to estimate transition probabilities between river sections, to determine whether brook trout: (a) moved between multiple river sections and (b) entered lacustrine habitats as seasonal refuges, but eventually returned to lotic habitat. We also used a Cormack-Jolly-Seber model to evaluate whether apparent survival of brook trout in the WBWR was comparable to other populations. Few fish moved among river sections or used lacustrine habitat (<5% of tagged fish); most brook trout remained in sections where they were initially tagged, potentially due to quality habitat located throughout the river. Like other studies, brook trout in the WBWR appear to experience high mortality based on low number of detections, few physical recaptures and an estimated eight-month apparent survival rate of 0.27. In scenarios where fish can already access suitable habitat, removal of dams may not result in substantial increases in fish movement and colonisation of newly accessible habitat may not occur immediately.     

Hormonal changes during meiotic maturation and ovulation in the brook trout (Salvelinus fontinalis)

The plasma levels of estradiol-17 (E2), 17, 20-dihydroxy-4-pregnen-3-one (17,20-P) and gonadotropin (GTH) were measured in brook trout (Salvelinus fontinalis) during the period from the end of vitellogenesis to postovulation. Blood samples were taken according to specific stages of maturation, including germinal vesicle breakdown (GVBD) and ovulation. E2 levels were quite high (45 ng/ml) at the end of vitellogenesis (and prior to GVBD) and dropped precipitously by GVBD (2 ng/ml). They remained low through ovulation and postovulation. 17,20-P levels were low prior to GVBD (0.7 ng/ml) and increased dramatically at GVBD (148 ng/ml). The levels of 17,20-P remained high at ovulation (142 ng/ml) and then dropped significantly within 24 h to approximately half of the ovulatory values. They decreased even further by 7 days postovulation. GTH levels rose gradually through GVBD and ovulation from a postvitellogenic level of approximately 3 ng/ml to a 7 day postovulatory value of approximately 10 ng/ml. The overall results; 1) decrease in estradiol prior to GVBD, 2) increase in 17,20-P at GVBD and 3) gradual GTH rise through GVBD and ovulation, are similar to those reported for other salmonids.     

The purification and development of a quantitative enzyme linked immunosorbent assay (ELISA) for the measurement of vitellogenin in diploid and triploid brook trout (Salvelinus fontinalis)

Vitellogenin (VTG) was purified from the plasma of estradiol-17β (E₂) treated male brook trout (Salvelinus fontinalis; bt) using gel filtration and anion exchange chromatography. An antiserum to bt-VTG was raised in rabbits, then used to detect bt-VTG by Western blot analysis. Purified bt-VTG and its antibody were then used to develop an antibody-capture, competitive enzyme-linked immunosorbant assay (ELISA). Confirmation of the purified protein as bt-VTG was based on the characteristics of high molecular weight (∼562 kDa), dominant plasma protein in vitellogenic females and induction by exogenous E₂ treatment in males. The antiserum recognized a major 184 kDa polypeptide as well as minor bt-VTG degradation products (150–66 kDa) in purified bt-VTG and in plasma from vitellogenic females. Low levels of antibody cross reactivity were shown with plasma from nonvitellogenic females, control rabbit serum, and several other antisera known to not contain VTG. The ELISA had a minimum detection limit of 8 ng ml⁻¹ and intra- and inter-coefficients of variation less than 9% and 15%, respectively. The ELISA demonstrated parallel binding slopes among dilution curves of purified bt-VTG standard and plasma from diploid and triploid brook trout females. Using the ELISA, maximum plasma VTG levels of 93.5±33.6 mg ml⁻¹ were detected in vitellogenic diploid females, whereas only 0.18±0.15 mg ml⁻¹ were detected in triploid females of the same age (n=5 for each). Diploid and triploid males cohabitating with vitellogenic females showed measurable levels of plasma VTG during vitellogenesis in females (i.e., 0.17 and 0.06 mg ml⁻¹, respectively (n=1)). This revised version was published online in August 2006 with corrections to the Cover Date.     

Hemoglobin and subunit multiplicity in the rainbow trout (Oncorhynchus mykiss) hemoglobin system

Hemoglobin (Hb) multiplicity is a commonly used index of phylogenetic differentiation and molecular adaptation in fish. Despite improvements in technologies for characterizing protein structure, our knowledge of the component composition of the Hb system in rainbow trout, Oncorhychus mykiss, has remained almost unchanged through three decades. The Hb is considered to consist of four components, of which two (HbI and HbIV) have been extensively characterised with regard to structure and function. Using a variety of molecular techniques we demonstrate the presence of at least nine different Hb fractions composed of nine (five α and four β globin) chains, and that HbI and HbIV consist of two and five individual components, respectively. These findings indicate that the published data on trout HbI and HbIV refer to mixtures of isoHbs and need to be reappraised. This revised version was published online in August 2006 with corrections to the Cover Date.     

Patterns of growth and nutrient deposition in lake trout (Salvelinus namaycush), brook trout (Salvelinus fontinalis) and their hybrid, F1 splake (Salvelinus namaycush × Salvelinus fontinalis) as a function of water temperature

Several fish species of the genus Salvelinus are used for stocking freshwaters in North and South America and much of Europe but there is little information about their growth and carcass composition. Lake trout (S. namaycush), brook trout (S. fontinalis) and their hybrid F₁ splake (S. namaycush × S. fontinalis) (initial body weight ca. 2–4 g) were raised at 6.4, 10.6 and 14.9 °C to examine growth and nutrient deposition as a function of water temperature. In all species, weight gain and feed intake increased significantly with water temperature and feed efficiency was significantly lower at 6.4 than at 10.6 and 14.9 °C. In brook trout, Thermal‐unit Growth Coefficient (TGC) growth rate was significantly lower at 6.4 than at 10.6 and 14.9 °C, while in F₁ splake TGC was only lower at 6.4 than 10.6 °C. Expressed in terms of relative composition a significant effect of temperature was observed. In all species, moisture content decreased while crude protein, lipid, ash and energy contents increased with increased temperature. Expressed in absolute terms, however, a significant effect of temperature was not observed. In all species carcass contents increased significantly with increased live body weight and were best described by simple linear equations. Gross energy concentration was significantly affected by both water temperature and body weight. These data indicate that the growth of these species is a function of water temperature and, in absolute terms, carcass composition is mainly a function of body weight and not water temperature. Also, such simple linear equations bode well for modification of existing feed requirement and waste outputs models; improving their applicability to these species.     

Loma salmonae (Putz, Hoffman and Dunbar, 1965) in the rainbow trout, Salmo gairdneri Richardson, and L. fontinalis sp. nov. (Microsporida) in the brook trout, Salvelinus fontinalis (Mitchill)

Abstract. Loma salmonae (Putz, Hoffman and Dunbar, 1965), Morrison & Sprague, 1981, in the rainbow trout, Salmo gairdneri Richardson, was studied with the electron microscope. It is morphologically very similar to L. morhua Morrison and Sprague, 1981, in the Atlantic cod, Gadus morhua L., having only minor differences. Loma sp. Morrison and Sprague, 1981, in the brook trout, Salvelinus fontinalis (Mitchill), is more like L. salmonae than the other species of Loma , but was deemed to be distinct. It was named L. fontinalis sp. nov.