Early Life Stage Survival and Susceptibility of Brook Trout,Coho Salmon,Rainbow Trout,and Their Reciprocal Hybrids to Infectious Hematopoietic Necrosis Virus

Abstract

Triploid (heat-shocked) and diploid groups of rainbow trout Oncorhynchus mykiss, brook trout Salvelinus fontinalis, coho salmon Oncorhynchus kisutch, and reciprocal hybrids were produced, monitored for early life stage survival, and evaluated for susceptibility to infectious hematopoietic necrosis virus (IHNV). The female rainbow trout × male brook trout triploid hybrids had significantly greater (P < 0.01) survival than the diploid hybrids of this cross. The heat-shocked hybrid group of the female rainbow trout × male coho salmon also exhibited significantly greater survival to the eyed egg stage of development than the untreated group of this hybrid. Studies of the susceptibility of treatment groups to a 1990 IHNV isolate from the Hagerman Valley were conducted by using a standardized immersion exposure procedure at one or two different mean body weights. The diploid brook trout and coho salmon and two triploid hybrids (female rainbow trout × male brook trout or male coho salmon) were significantly less (P < 0.05) susceptible to IHNV than the pure-species diploid and triploid rainbow trout groups.     

Stress Protein Expression in Coho Salmon with Bacterial Kidney Disease

Abstract

The expression of stress protein-70 (SP-70), also known as heat shock protein-70 (HSP-70), was measured by enzyme-linked immunosorbent assay in kidney and liver from coho salmon Oncorhynchus kisutch with bacterial kidney disease (BKD) experimentally induced by injection with Renibacterium salmoninarum. Fish with BKD had more SP-70 in both kidney and liver than did the control fish. The SP-70 measured was derived from the host tissue, not from the pathogen. In fish without clinical disease, SP-70 was not significantly elevated. Elevated stress protein (SP) levels in fish with BKD raise the possibility that BKD or other infectious diseases may interfere with the use of SP induction as a marker of environmental stress in fish and lead to statistical artifacts when the prevalence of disease is not considered.     

Investigation of the Link between Broodstock Infection,Vertical Transmission,and Prevalence of Flavobacterium psychrophilum in Eggs and Progeny of Rainbow Trout and Coho Salmon

Abstract

The etiological agent of bacterial coldwater disease (BCWD), Flavobacterium psychrophilum, can be transmitted both vertically and horizontally. Outbreaks of BCWD can result in significant losses in salmonid aquaculture. Reduction of outbreaks in fry may be possible through implementation of a management strategy in which progeny of heavily infected broodstock are culled from the general population. Diagnostic assays to quantify F. psychrophilum concentrations in tissue samples and confirm presence of the bacterium in ovarian fluid have been previously validated. In the current study, these assays were used to screen 60 female Rainbow Trout Oncorhynchus mykiss and 60 female Coho Salmon O. kisutch broodstock at two aquaculture facilities. Eyed eggs from 10 female broodstock (five fish from each facility) exhibiting graded levels of infection were transferred to the University of Idaho and monitored through early life stages for the presence of F. psychrophilum. Female Rainbow Trout broodstock were not positive for F. psychrophilum by enzyme-linked immunosorbent assay (ELISA) and prevalence was low in these progeny. However, ELISA optical density values for kidney correlated to F. psychrophilum prevalence in progeny (r = 0.938, P < 0.05) of Coho Salmon. Nested PCR on ovarian fluid was not a reliable indicator of vertical transmission in either species as broodstock ovarian fluid results did not correlate to F. psychrophilum prevalence in eyed eggs. Further research with these assays is necessary; however, results from this study indicate that broodstock screening may be a potential tool for evaluating F. psychrophilum infection levels, which could become an important component for disease management.

Received December 19, 2012; accepted December 8, 2013     

Epizootiological Study of Bacterial Cold-Water Disease in Pacific Salmon and Further Characterization of the Etiologic Agent,Flexibacter psychrophila

Abstract

Isolates of Flexibacter psychrophila were obtained from chinook salmon Oncorhynchus tshawytscha and coho salmon O. kisutch that had previously sustained epizootics of coldwater disease. The pathogen was readily isolated from kidney and mucus of convalescent fish. The organisms were relatively inert in most standard microbiological media but were structurally and serologically homogenous by examination of whole cell protein lysates by sodium dodecyl sulfate polyacrylamide gel electrophoresis. In contrast to the homogeneity observed in phenotypic and serologic assays, the isolates studied elaborated varied ribotypes. All isolates produced a single rDNA spacer amplification product of about 240 base pairs.     

Differing Resistance of Atlantic Salmon Strains and Rainbow Trout to Gyrodactylus salaris Infection

Abstract

Laboratory studies were conducted on the susceptibility of different strains of Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss to infection with the monogenean Gyrodactylus salaris. This parasite, probably originating from the Baltic Sea region, is known to minimally affect Neva River (Baltic Sea) Atlantic salmon. However, following its introduction into Norway, G. salaris has caused severe mortality and morbidity among Norwegian Atlantic salmon, which are considered a highly susceptible strain. The cohabitation experiment included one stock of rainbow trout and four different strains of Atlantic salmon from the Baltic Sea region (Mörrum River, Sweden), Europe (Skjern River, Denmark; Conon River, Scotland), and North America (Bristol Cove River, Canada). Fish were exposed to a Norwegian strain of G. salaris, and parasite population development and distribution were monitored for 7 weeks. Rainbow trout exhibited low susceptibility to G. salaris infection, whereas Conon River and Skjern River Atlantic salmon were highly susceptible and exhibited high mortality rates. Mörrum River Atlantic salmon exhibited intermediate susceptibility and low mortality. Bristol Cove River Atlantic salmon harbored relatively low parasite numbers, but fish mortality was high. Our experiment showed that the Danish Skjern River strain of Atlantic salmon is highly susceptible to G. salaris infection, further supporting the hypothesis that Atlantic Ocean strains are more susceptible to G. salaris infection than are Baltic strains.     

Monoclonal Antibody-Based Analysis of the Renibacterium salmoninarum p57 Protein in Spawning Chinook and Coho Salmon

Abstract

Renibacterium salmoninarum, causative agent of bacterial kidney disease of salmonid fish, produces large amounts of soluble proteins during infection and broth culture. An enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibodies was developed for the precise quantification of p57, a major component of these proteins. Kidney, spleen, blood, and reproductive fluids of adult Pacific salmon Oncorhynchus spp. were examined by means of the assay. Kidney and spleen harbored the highest concentrations of this antigen. In the populations of returning salmon tested, 5–25% of the fish had p57 concentrations above a baseline level of 3 ng antigenig tissue, and antigen concentrations as high as 200 μg/g tissue were detected in kidneys of individual fish. The ELISA was compared to direct fluorescent antibody analysis, in which rabbit anti-R. salmoninarum antiserum was used to identify infected fish. There was 99% agreement (199 of 201 examined fish) between the two methods. Western blot analysis was used to demonstrate that antigenic proteins present in infected fish were similar to those seen from antigen prepared from broth culture of R. salmoninarum, although less degradation of p57 appears to occur in vivo.     

Observations on the Occurrence of Bacterial Gill Disease and Amoeba Gill Infestation in Rainbow Trout Cultured in a Water Recirculation System

Abstract

Spontaneous outbreaks of bacterial gill disease (BGD) occurred in 17-44-g rainbow trout Oncorhynchus mykiss after they were placed in a water recirculation system. In each of five groups stocked from September 1991 through July 1992, BGD occurred within 6–8 d after stocking. In each instance, BGD was followed by a secondary amoeba infestation. The spontaneous BGD outbreaks did not occur among previously stocked groups that had recovered from earlier BGD disease outbreaks. Examination of gill tissue by Gram stain and indirect fluorescent antibody technique showed increased numbers of filamentous bacteria associated with BGD after the rainbow trout were stocked into the system. Bacterial numbers decreased after a 1-h treatment with chloramine-T at concentrations of 9–15 mg/L but increased within 2 d after treatment. Although the chloramine-T treatments controlled mortality related to BGD, the amoeba infestation persisted. Histological examination of gills showed some focal hyperplasia before the rainbow trout were placed into the recirculation system, but hyperplasia became more extensive and lamellar fusion and mild telangiectasis developed within a week after placement in the system. The density at which the fingerling rainbow trout were stocked and the suspended solids present in tank water may have contributed to the BGD outbreaks. Exposure of juvenile rainbow trout to tank water from the recirculation system before they were placed into the system did not afford them protection against BGD after stocking.     

Temperature Related Absorption and Excretion of Sulphadimidine in Rainbow Trout,Salmo Gairdneri

The main purpose of the present study was to see if a kinetic model and a computer program for characterizing the rate of uptake and clearance of chemicals in aquatic organisms (Biofac, OECD’s Chemicals Testing Programme) was applicable to pharmacokinetic studies in fish. The program seems suitable for such studies.The effect of temperature upon rate of absorption of sulphadimidine from medicated water and elimination of the same drug was investigated in rainbow trout. A rise in the temperature from 7° C to 14° C more than doubled the calculated rate constants of absorption and elimination. The biological half life (t_½ β) and the time to reach 90 % of steady state (t_{90% ss}) at 14° C were approximately half the values at 7° C.     

Regulatory Effects of Water Temperature on Loma salmonae (Microspora) Development in Rainbow Trout

Abstract

Water temperature, a pivotal factor influencing interactions between teleosts and pathogens, was examined to determine its effects on the kinetics of xenoma formation and dissolution subsequent to experimental exposure of rainbow trout Oncorhynchus mykiss to the microsporidian gill pathogen Loma salmonae. The permissive water temperature range in which xenomas developed was between 9° and 20°C. Parasite development was arrested at temperatures outside this range, as indicated by the absence of visible xenomas among exposed fish. In addition, when these trout were subsequently moved to temperatures within the permissive range, xenomas failed to develop. Water temperature, within the permissive range, had no significant effect on either the number of xenomas that formed or the proportion of fish that developed xenomas following gastric intubation with a standard dose of spores. The relationship between water temperature and xenoma onset-time was best described (R ² = 88.3%) by polynomial regression analysis: onset = 320 ? 33.4T + 0.9547T ², where T is temperature (°C). Xenoma onset rate was also described through a modified degree-days model, yielding a predictive equation appropriate for use under conditions of fluctuating temperature. The thermal units, expressed as days × (°C above 7°C) necessary for xenoma onset were 298.6 on average. Xenoma dissolution rates, from the time of onset, also appeared to have a trend; more rapid dissolution occurred as temperatures increased. However, this trend correlated minimally with regression models.     

养殖虹鳟与大西洋鲑肉质和营养成分比较研究

试验选取10尾贵州都匀养殖场的虹鳟(1562±32)g和10尾市场出售的挪威大西洋鲑(1856±45 g),采集其背部肌肉,分别进行质构特性及营养成分分析。结果表明:质构特性上,虹鳟肌肉的失水率显著低于大西洋鲑(P<0.05),剪切力显著高于大西洋鲑(P<0.05);常规成分中,虹鳟肌肉的粗脂肪含量显著低于大西洋鲑(P<0.05),二者的粗蛋白含量无显著差异(P>0.05);脂肪酸组成上,虹鳟肌肉的n-6多不饱和脂肪酸含量显著低于大西洋鲑(P<0.05),但基于脂肪酸组成获得的饱和度(S/P)、动脉粥样硬化指数(AI)、凝血指数(TI)和降胆固醇/致高血脂比率(HH)等指标无显著性差异(P>0.05);氨基酸组成方面,虹鳟肌肉内的人体必需氨基酸比例显著低于大西洋鲑(P<0.05),鲜味氨基酸含量显著高于大西洋鲑(P<0.05)。结果表明,养殖虹鳟在肉质质构上相比大西洋鲑较“干硬”,且虹鳟肌肉多不饱和脂肪酸和必需氨基酸含量均低于大西洋鲑。     

Effect of Nanophyetus salmincola and Bacterial Co-Infection on Mortality of Juvenile Chinook Salmon

The freshwater trematode Nanophyetus salmincola has been demonstrated to impair salmonid immune function and resistance to the marine pathogen Vibrio anguillarum, potentially resulting in ocean mortality. We examined whether infection by the parasite N. salmincola similarly increases mortality of juvenile Chinook Salmon Oncorhynchus tshawytscha when they are exposed to the freshwater pathogens Flavobacterium columnare or Aeromonas salmonicida, two bacteria that juvenile salmonids might encounter during their migration to the marine environment. We used a two-part experimental design where juvenile Chinook Salmon were first infected with N. salmincola through cohabitation with infected freshwater snails, Juga spp., and then challenged with either F. columnare or A. salmonicida. Cumulative percent mortality from F. columnare infection was higher in N. salmincola-parasitized fish than in nonparasitized fish. In contrast, cumulative percent mortality from A. salmonicida infection did not differ between N. salmincola-parasitized and nonparasitized groups. No mortalities were observed in the N. salmincola-parasitized-only and control groups from either challenge. Our study demonstrates that a relatively high mean intensity (>200 metacercariae per posterior kidney) of encysted N. salmincola metacercariae can alter the outcomes of bacterial infection in juvenile Chinook Salmon, which might have implications for disease in wild fish populations.

Received February 24, 2015; accepted September 7, 2015     

Case Report: Strawberry Disease in Farmed Chilean Rainbow Trout

Strawberry disease is a chronic, nonlethal skin condition that affects Rainbow Trout Oncorhynchus mykiss in the United States and several European countries, where it is also known as red-mark syndrome. We provide the first identification and characterization of three strawberry disease outbreaks occurring at two aquaculture farms in southern Chile. Clinically affected fish weighing an average of 400 g presented multiple bright-red, usually raised, skin lesions on the flank, ventral surface, and dorsal surface. A PCR using Rickettsia-like-organism (RLO)-specific primers was performed on nine affected fish, and all skin samples were positive for the RLO 16S ribosomal RNA sequence. All PCR results for Flavobacterium psychrophilum and other bacterial and viral pathogens were negative. Histopathological examination of the skin lesions revealed extensive dermatitis, with severe lymphocytic infiltration in advanced cases. This report is the first to describe strawberry disease in farmed Chilean Rainbow Trout. Additional studies are needed to evaluate the risk for Rainbow Trout culture; fish challenge experiments should be performed to fulfill Koch’s postulates and to demonstrate that RLO is the cause of this disease.

Received December 27, 2014; accepted October 23, 2015.     

Ichthyophoniasis: An Emerging Disease of Chinook Salmon in the Yukon River

Abstract

Before 1985, Ichthyophonus was unreported among Pacific salmon Oncorhynchus spp. from the Yukon River; now it infects more than 40% of returning adult Chinook salmon O. tshawytscha. Overall infection prevalence reached about 45% in the Yukon River and about 30% in the Tanana River between 1999 and 2003. Mean infection prevalence was greater in females than males in the main-stem Yukon River during each of the 5 years of the study, but the infection prevalence in males increased each year until the difference was no longer significant. Clinical signs of ichthyophoniasis (presence of visible punctate white lesions in internal organs) were least at the mouth of the Yukon River (～10%) but increased to 29% when fish reached the middle Yukon River and was 22% at the upper Tanana River. However, clinical signs increased each year from 7% in 1999 to 27% in 2003 at the mouth of the river. As fish approached the upper reaches of the Yukon River (Canada) and the spawning areas of the Chena and Salcha rivers (Alaska), infection prevalence dropped significantly to less than 15% in females on the Yukon River and less than 10% for both sexes in the Chena and Salcha rivers, presumably because of mortality among infected prespawn fish. Age was not a factor in infection prevalence, nor was the position of fish within the run. The source of infection was not determined, but Ichthyophonus was not found in 400 Pacific herring Clupea pallasi from the Bering Sea or in 120 outmigrating juvenile Chinook salmon from two drainages in Alaska and Canada. Freshwater burbot Lota lota from the middle Yukon River were subclinically infected with Ichthyophonus, but the origin and relationship of this agent to the Chinook salmon isolate is unknown.     

Gill Infection Model for Columnaris Disease in Common Carp and Rainbow Trout

Challenge models generating gill lesions typical for columnaris disease were developed for the fry of both Common Carp Cyprinus carpio and Rainbow Trout Oncorhynchus mykiss by means of an immersion challenge and Flavobacterium columnare field isolates were characterized regarding virulence. Carp inoculated with highly virulent isolates revealed diffuse, whitish discoloration of the gills affecting all arches, while in trout mostly unilateral focal lesions, which were restricted to the first two gill arches, occurred. Light microscopic examination of the gills of carp exposed to highly virulent isolates revealed a diffuse loss of branchial structures and desquamation and necrosis of gill epithelium with fusion of filaments and lamellae. In severe cases, large parts of the filaments were replaced with necrotic debris entangled with massive clusters of F. columnare bacterial cells enwrapped in an eosinophilic matrix. In trout, histopathologic lesions were similar but less extensive and much more focal, and well delineated from apparently healthy tissue. Scanning and transmission electron microscopic observations of the affected gills showed long, slender bacterial cells contained in an extracellular matrix and in close contact with the destructed gill tissue.

This is the first study to reveal gill lesions typical for columnaris disease at macroscopic, light microscopic, and ultrastructural levels in both Common Carp and Rainbow Trout following a challenge with F. columnare. The results provide a basis for research opportunities to examine pathogen–gill interactions.

Received January 10, 2014; accepted July 27, 2014     

Blood Parameters and Immune Status of Rainbow Trout with Proliferative Kidney Disease

Abstract

Blood parameters, disease resistance, and the immune response were sequentially evaluated in rainbow trout Oncorhynchus mykiss with proliferative kidney disease (PKD). The fish were maintained under laboratory conditions, and the study group went through a full cycle of the disease. Hematological and serological changes occurred primarily in those fish with severe kidney lesions. Fish infected with the parasite that causes PKD demonstrated a greater resistance to bacterial challenge, and their immune responses were heightened when compared with those of uninfected fish. These data suggest that PKD alone is not a predisposing factor for secondary infections if the fish does not incur severe renal lesions.     

The Impact of Egg Ozonation on Hatching Success,Larval Growth,and Survival of Atlantic Cod,Atlantic Salmon,and Rainbow Trout

The direct exposure of fish eggs to ozonated water has generated interest as a means of ensuring pathogen-free eggs without the use of harsh chemicals. However, there are numerous knowledge gaps, including safe contact times, exposure levels, and potential long-term effects on aquaculture species in both freshwater and seawater. The effect of different ozone (O₃) doses (0.5–1.0, 1.5–2.0, and 2.5–3.0 mg of O₃/L for 90 s) on recently fertilized eggs of Atlantic Cod Gadus morhua and eyed eggs of Atlantic Salmon Salmo salar and Rainbow Trout Oncorhynchus mykiss was evaluated in comparison with the effects of two commercial disinfectants: Perosan (0.004 mg/L) and Ovadine (100 mg/L). The impact of ozone application was evaluated based on hatching success, larval nucleic acid concentration, larval growth, and survival. Overall, results indicated that ozonation of Atlantic Cod eggs at a dose less than 3.0 mg/L for 90 s produced no negative effect on the larvae up to 30 d posthatch. Furthermore, ozonation of Atlantic Salmon and Rainbow Trout eggs generated no negative effect on the larvae, based on monitoring until 85% yolk sac re-absorption (16 d posthatch).

Received May 6, 2014; accepted October 24, 2014     

Branchial Ionic Flux Responses in Rainbow Trout to Chloramine-T after Acclimation to Different Levels of Water Hardness

Abstract

Whole-body unidirectional and net Na⁺ and Cl^– fluxes were determined in rainbow trout Oncorhynchus mykiss exposed to chloramine-T (9 mg/L) alone or in combination with 17 mmol NaCl/L (0.1 % weight per volume, w/v) after acclimation to either city of Ottawa tap water (TW: Na⁺ 0.121 ± 0.004 mmol/L; K⁺ 0.015 ± <0.001 mmol/L; Ca²⁺ 0.351 ± 0.004 mmol/L; Cl^– 0.141 ± 0.002 mmol/L) or artificial soft water (ASW: Na⁺ 0.043 ± 0.003 mmol/L; K⁺ 0.005 ± <0.001 mmol/L; Ca²⁺ 0.063 ± 0.002 mmol/L; Cl^– 0.026 ± 0.002 mmol/L). Preexposure fluxes revealed that ASW-acclimated fish had reduced rates of Na⁺ uptake and efflux compared with TW-acclimated fish. However, rates of Cl^– uptake among ASW-acclimated fish were comparable with those of TW-acclimated fish. Exposure to chloramine-T in TW resulted in significant increases in the Cl^– efflux, giving rise to a significant and negative net flux. Exposure of ASW-acclimated fish to chloramine-T caused a significant negative increase in the Na⁺ efflux and net flux but only a negative increase in the Cl^– net flux. The addition of 17 mmol NaCl/L (0.1% w/v) eliminated the negative net ionic fluxes, resulting in significant increases in influx. The combined use of chloramine-T with NaCl is a potential method of remediating ionic disturbances that may be caused by treating trout with chloramine-T.     

Cellular Inflammatory Response of Rainbow Trout to the Protozoan Parasite that Causes Proliferative Kidney Disease

Abstract

The cellular inflammatory response of rainbow trout Oncorhynchus mykiss (formerly Salmo gairdneri) to the myxozoan parasite PKX that causes proliferative kidney disease was investigated. The response was studied from 3 to 20 weeks after the fish were injected with infected kidney homogenate. Kidney samples were examined by light and electron microscopy. In contrast to most myxosporeans, PKX provoked a severe host response. Parasites were found in peritubular capillaries and sinusoids 3 weeks postinjection. The initial response to PKX was hemopoietic hyperplasia followed by a marked granulomatous nephritis that was resolved by termination of the study at 20 weeks postinjection. The macrophage was the predominant cell type involved in the inflammatory response to PKX. We presume that the macrophage effectively interrupts the development of PKX and eliminates the parasite from the host.     

Effects of Supplemental Amino Acid-Chelated Trace Elements on the Immune Response of Rainbow Trout Subjected to Bacterial Challenge

Abstract

This study examined whether supplemental amino acid-chelated trace elements were effective in improving disease resistance in rainbow trout Oncorhynchus mykiss. Four semipurified diets containing trace elements from either sulfates or an amino acid chelate source were fed to the fish for 18 weeks. Subsequently, fish weighing about 20–40 g were challenged with β-hemolytic Streptococcus sp. The cumulative mortality was less in the chelate-supplemented groups but not significantly different from that of the sulfate groups. Antibody titer was significantly higher in the chelate-fed fish. Lysozyme activity and total immunoglobulin content were slightly more enhanced in the chelate-fed than in the sulfate-fed groups. Despite the increased antibody production, no distinct improvement in disease resistance against Streptococcus sp. was achieved by supplementing rainbow trout diets with amino acid-chelated trace elements. Further confirmatory studies focusing on the amounts of the trace elements used are needed to shed light on the influence of amino acid chelates on disease resistance.     

Survival Rates of Out-Migrating Yearling Chinook Salmon in the Lower Columbia River and Plume after Exposure to Gas-Supersaturated Water

In 2011, unusually high flows caused total dissolved gas (TDG) levels in the Columbia River, USA, to escalate well above the 120% regulatory limit that was imposed to prevent harmful impacts to aquatic organisms. After observing gas bubble trauma (GBT) in dead yearling Chinook Salmon Oncorhynchus tshawytscha (smolts) held in tanks, we compared estimated survival rates of acoustic-tagged in-river-migrating (IR) and transported (TR) smolts that were released below Bonneville Dam prior to and during the period of elevated TDG (>120%). The log odds of estimated daily survival in the lower river and plume was significantly lower for IR smolts that were released during elevated TDG (maximum possible exposure = 134%) than for IR smolts released when TDG was less than 120%. The TR smolts that were released 10–13 km below Bonneville Dam during elevated TDG had lower maximum possible exposure levels (126% TDG), and the log odds of estimated daily survival in the lower river and plume did not differ from that of TR smolts released when TDG was less than 120%. Direct mortality due to GBT is probably reduced in natural settings relative to laboratory experiments because smolts can move to deeper water, where pressure keeps gasses in solution, and can migrate downstream of the spillway, where TDG levels decrease as the river returns to equilibrium with the atmosphere. However, initially nonlethal GBT may reduce survival rates by increasing smolt susceptibility to predation and infection. Although our findings are limited by the observational nature of the study, our analysis is the first direct assessment of gas supersaturation’s potential influence on survival of free-ranging smolts in the river and coastal ocean below a large dam. Experiments using simultaneous releases of control and gas-exposed groups are warranted and should consider the possibility that the chronic effects of TDG exposure on survival are important and persist into the early marine period.

Received October 5, 2015; accepted August 18, 2016 Published online October 24, 2016