Pathogenesis of infectious pancreatic necrosis virus infection in brook trout, Salvelinus fontinalis (Mitchill), following intraperitoneal injection

Abstract. The pathogenesis of IPNV infection was examined in young brook trout following intraperitoneal inoculation. Fish were sampled sequentially and the distribution of virus determined by virus isolation, histopathology and immunofluorescence. Following intraperitoneal inoculation, virus entered the peritoneal cavity and by 2 days post-inoculation virus had interacted with the pancreatic exocrine cells. Replication in these cells resulted in the production of IPNV-specific antigen, necrosis of infected cells, and spread of infectious virus to adjacent tissue. Areas of viral replication were at first multifocal but tended to merge as they grew. Eventually, most of the acinar tissue became involved and only small pockets of normal acinar cells remained. Twelve-week old brook trout survived with only a small amount of functional exocrine pancreas and limited virus replication continued in this tissue for weeks. Extensive virus replication appeared to take place only in the pancreas although viral antigen and slight pathological changes were found in the renal interstitium and livers of some fish.     

Relevance of Renibacterium salmoninarum in an asymptomatic carrier population of brook trout, Salvelinus fontinalis (Mitchill)

Abstract. These studies were done to determine the prevalence of infection, over time, of progeny from a population of 1988 year class brook trout, Salvelinus fontinalis (Mitchill), that was asymptomatically infected with Renibacterium salmoninarum , the causative agent of bacterial kidney disease. This population and its progeny from the 1991 and 1992 spawnings were monitored for approximately 3 years for prevalence of R. salmoninarum and development of bacterial kidney disease. Ovarian fluid samples from the 1991 and 1992 spawnings, and from a 1993 spawning of the 1991 progeny, were analysed for R. salmoninarum by enzyme linked immunosorbent assay (ELISA) and the membrane filtration fluorescent antibody technique. Kidney and spleen tissues of the 1988 year class were assayed by ELISA following the 1992 spawning. The progeny of the 1991 and 1992 spawnings were followed for increased prevalence of R. salmoninarum by the ELISA. Kidney tissues were assayed and sampling began once the fish were large enough. Each population of progeny was sampled quarterly for one year. Overall, the progeny displayed decreasing prevalence of R. salmoninarum and all ovarian fluids of the 1991 progeny (in 1993) were negative. Brood fish and progeny remained asymptomatic during the course of the study.     

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.     

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.     

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.     

Food availability and growth of the brook trout, Salvelinus fontinalis (Mitchill), in a French Alpine lake

Plan Vianney lake, situated in the Oisans massif (French Alps) at an altitude of 2250 m, has a fish fauna composed of brook trout, Salvelinus fontinalis (Mitchell), and Artic charr, Salvelinus alpinus (L.). The present study examines the feeding behaviour of S. fontinalis in 1993–94 but also gives some data on S. alpinus . The brook trout represented 80% of the fish fauna of the lake. Dietary analysis showed that this species is an opportunistic feeder and revealed similarities in the trophic niches of S. fontinalis and S. alpinus and a low level of potentially available food resources. The annual growth of S. fontinalis was short and the growth rate was slow compared with other populations. Under these extreme conditions, female fish required several summers to replenish their reserves of energy and were only in condition to spawn once every 2 or 3 years. Although the fecundity of the population was low, it was sufficient to maintain the species in the lake which is not exploited.     

Association between initial weight and weight gain in brook trout, Salvelinus fontinalis Mitchill, and brown trout, Salmo trutta L.

Abstract. Individual weighings of samples of brook trout, Salvelinus fontinalis Mitchill, and brown trout, Salmo trutta L., al the initiation and termination of their growth period, enabled estimating the association of weight gain on initial weight. In both cases the regressions were positive and the correlations significant. Variation in initial weight among tested individuals and groups is expected to bias growth estimates of these groups in programmes of genetic testing, as a result of this association. Obviating this bias requires an adequate correcting technique, involving the estimation of an environmentally generated correction factor. Phenotypic regression coefficients, as estimated in this investigation, cannot serve as correction factors, but may be regarded as maximum estimates.     

Cytogenetic and histological studies of the brook trout, Salvelinus fontinalis (Mitchill), and the Arctic char, S. alpinus (L.) hybrids

Although brook trout and the Arctic char hybrids are able to reproduce, individuals with decreased fertility or even fish that are unable to produce any gametes have been also described. Abnormal gonadal development and disturbances in the gamete production in the char hybrid offspring may be triggered by the odd chromosome number and disturbances in their pairing during meiosis. To verify this hypothesis, cytogenetic examination and the gonadal histology analysis of the brook trout x Arctic char hybrids were carried out. Diploid chromosome number in the studied char (F₁) hybrids varied from 82 to 84 (FN = 99–102). Among 28 hybrids, 12 males, three females, nine intersex individuals and two sterile specimens were described. In the case of two individuals, gonads were not found. Diploid chromosome numbers in the males and intersex individuals varied from 82 to 84. Chromosome numbers in the females were 82 and 83 chromosomes. Two sterile fish exhibited karyotypes composed of 82 and 84 chromosomes. Predominance of the ovarian component in the intersex gonads and gonadal sex ratio distortion towards the males suggested hybrid females had problems with gonadal differentiation. However, the lack of the clear relationship between chromosome number and gonadal development in the studied hybrids did not support our hypothesis that odd chromosome number may be responsible for such reproductive disturbances in the hybrid individuals. We have presumed that sterility and intersexual development of the gonads may be caused by interactions between brook trout and Arctic char genes on the sex chromosomes and autosomes rather than unpairing of the parental chromosomes.     

Immunization of adult brook trout, Salvelinus fontinalis (Mitchill), fails to prevent the infectious pancreatic necrosis virus (IPNV) carrier state

Abstract. Adult brook trout, Salvelinus fontinalis (Mitchill), mounted a strong humoral immune response after injection with inactivated infectious pancreatic necrosis virus (IPNV) in Freund's complete adjuvant (FCA). However, this immunization did not prevent the fish from becoming IPNV carriers. After an injection challenge with virulent IPNV, the immunized and control fish (FCA or water) shed virus in the faeces and reproductive products and had IPNV-infected leucocytes and visceral organs. Initially, from 1 to 3 weeks post-challenge (wpc), immunized fish had a lower prevalence of infection and virus titres in the plasma, and fewer infected leucocytes than the control fish. Immunization did not prevent the eventual infection of the leucocytes; over 75% of the immunized and control fish had leucocyte-associated viraemia from 6 to 15 wpc. When the organs were tested at 15 wpc, the immunized fish showed fewer infected organs per fish, and a lower prevalence of infection and virus titres in individual organs than the control fish, but these differences were not significant. Immunized male and female fish shed IPNV in the reproductive products, suggesting that immunization of adult fish would not prevent vertical transmission of IPNV to progeny.     

Fry age and size effects on immersion immunization of brook trout, Salvelinus fontinalis Mitchell, against infectious pancreatic necrosis virus

Abstract. Brook trout, Salvelinus fontinalis Mitchell, fry were divided into eight age groups of 1–8 weeks post-hatch (wph) and immunized by a single direct immersion in formalin-inactivated infectious pancreatic necrosis virus ( ipnv ). After a direct-immersion ipnv challenge given 4 weeks later, only fry immunized at 2, 3 and 6wph showed protection. The relative per cent survival 60 days after ipnv challenge was highest in 2 and 3 wph fry (45–50%) and decreased as fish increased in age or size. The same response was obtained one year later when four age groups of fry, from 1 to 4wph, were immunized with a different serotype and dose of ipnv . The 2 and 3 wph fry had mean weights between 49 and 60mg at immunization. Killed vaccines administered by immersion have not previously been reported as inducing protection in salmonid fry of such low weights. Analysis of the growth of the fry suggests that protection against IPNV requires immunization in the eleutheroembryo phase, during the time of slow weight gain. This appears to be in direct contrast to the stage of ontogeny and weight growth rate required for successful immunization against the bacterial pathogen Vibrio anguillarum. Although immunization with two ipnv serotypes reduced mortalities from challenges with these same virus isolates, it did not prevent ipnv infection of fry in any age group.     

Sanguinicola fontinalis sp nov. (Digenea: Sanguinicolidae): a blood parasite of brook trout, Salvelinus fontinalis (Mitchill), and longnose dace, Rhinichthys cataractae (Valenciennes)

Abstract. Sanguinicola fontinalis sp. nov., from the branchial, renal and cardiac blood vessels of brook trout. Salvelinus fontinalis (Mitchill), and longnose dace, Rhinichthys cataractae (Valenciennes), is described. The new species most closely resembles, hut differs from, Sanguinicola davisi of steelhead and rainbow trout, Salmo gairdneri Richardson. The adult differs from all known North American species in having 29–33 lateral tufts of blunt spines. The snail host is Leptoxis (Mudalia) carinata (Bruquière, 1792). Lodged and migrating miracidia caused considerable gill damage in the brook trout, resulting in high mortality when the fish were moved. To remove the hazard, the most severely affected brook trout were destroyed and the ponds drained and disinfected. Adult worms from Pennsylvania brook trout and West Virginia longnose dace were studied, as well as infected snails.     

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.     

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.     

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.     

Movement of brook trout (Salvelinus fontinalis) in four small subalpine streams in northern Colorado

We studied the movement of brook trout (Salvelinus fontinalis) in four small streams in northern Colorado using mark-recapture methods and weirs. The recapture rates of marked adult trout were low for all streams, and large numbers of unmarked adult trout, apparently immigrants, were found each year. Significantly more trout, immigrated into sections that were experimentally modified by installing low log dams, which increased depth, pool volume and the amount of overhead cover. The number of immigrant and resident trout was significantly related to the amount of cover in the sections. Resident trout were larger than immigrants in all streams in the last year of sampling. Most mobile brook trout moved upstream during summer on the two streams where weirs were operated, and upstream migrants were significantly larger than downstream migrants on both streams. We suggest that a high degree of movement may be an adaptive response by brook trout to the heterogeneous nature of small mountain streams.     

Microhabitat use by southern brook trout (Salvelinus fontinalis) in a headwater North Carolina stream

Brook trout are the one of the only Salvelinus species native to eastern North America and range from Canada to Georgia. Very little is known, however, about the ecology of the southern form of this species. We quantified microhabitat use of southern brook trout in Ball Creek NC, a third‐order stream, during six seasonal samples (summer 2010, autumn 2010, spring 2011, summer 2011, autumn 2011 and spring 2012). In general, trout preferentially occupied deeper microhabitats with lower mean velocities and higher amounts of erosional substrata than were randomly available. Older trout (1+ and 2+) occupied deeper microhabitats with lower mean velocities than yearling trout. These microhabitats typically represent ‘plunge pools’. Southern brook trout also occupied focal point velocities that were statistically indistinguishable from optimal velocities calculated for rainbow trout in the same system and thus may chose microhabitats that maximise net energy gain. Southern brook trout are found in isolated populations, and management strategies should focus on the preservation of plunge pool habitat for conservation of this subspecies.     

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     

The influence of Gyrodactylus salaris Malmberg 1957 (Monogenea) on the epidermis of Atlantic salmon,Salmo salar L., and brook trout,Salvelinus fontinalis (Mitchill): experimental studies

The effect of Gyrodactylus salaris on the epidermal structure of hatchery-reared brook trout parr and Norwegian Atlantic salmon parr was evaluated. Both species were initially susceptible to this parasite, but whereas populations on salmon increased until the host died, brook trout responded to, and eventually eliminated, their infections. Salmon skin samples taken 14 days p.i. showed a reduced mucous cell concentration (less than 1500 cells mm^–2 on the head compared with 2000 cells mm^–2 in controls; the same trend was also seen in other sites) and the epidermis was thinner (48 μm compared with 60 μm on pectoral fins; similar trend seen in other sites) than in uninfected controls kept under identical conditions. Brook trout skin samples were taken 50 days p.i., when the hosts had responded to, and almost eliminated, their infections. No change was then observed in mucous cell density, while the number of epidermal cell layers and the epidermal thickness of brook trout had increased slightly following infection. These results are related to the nature of the host response, and the thinning of the epidermis and loss of mucous cells may in some way be related to the inability of Norwegian salmon parr to respond to the parasite.