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USGS2007

Molecular and biochemical analysis of rainbow trout LCK suggests a conserved mechanism for T-cell signaling in gnathostomes

Two genes were identified in rainbow trout that display high sequence identity to vertebrate Lck. Both of the trout Lck transcripts are associated with lymphoid tissues and were found to be highly expressed in IgM-negative lymphocytes. In vitro analysis of trout lymphocytes indicates that trout Lck mRNA is up-regulated by T-cell mitogens, supporting an evolutionarily conserved function for Lck in the signaling pathways of T-lymphocytes. Here, we describe the generation and characterization of a specific monoclonal antibody raised against the N-terminal domains of recombinant trout Lck that can recognize Lck protein(s) from trout thymocyte lysates that are similar in size (???57 kDa) to mammalian Lck. This antibody also reacted with permeabilized lymphocytes during FACS analysis, indicating its potential usage for cellular analyses of trout lymphocytes, thus representing an important tool for investigations of salmonid T-cell function.

Molecular Immunology↗

USGS2014

Assessing reproductive and endocrine parameters in male largescale suckers ( Catostomus macrocheilus ) along a contaminant gradient in the lower Columbia River, USA

Persistent organochlorine pollutants such as polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (p,p′-DDE), and polybrominated diphenyl ethers (PBDEs) are stable, bioaccumulative, and widely found in the environment, wildlife, and the human population. To explore the hypothesis that reproduction in male fish is associated with environmental exposures in the lower Columbia River (LCR), reproductive and endocrine parameters were studied in male resident, non-anadromous largescale sucker (Catostomus macrocheilus) (LSS) in the same habitats as anadromous salmonids having conservation status. Testes, thyroid tissue and plasma collected in 2010 from Longview (LV), Columbia City (CC), and Skamania (SK; reference) were studied. Sperm morphologies and thyrocyte heights were measured by light microscopy, sperm motilities by computer-assisted sperm motion analysis, sperm adenosine triphosphate (ATP) with luciferase, and plasma vitellogenin (VTG), thyroxine (T4), and triiodothyronine (T3) by immunoassay. Sperm apoptosis, viability, mitochondrial membrane potential, nuclear DNA fragmentation, and reproductive stage were measured by flow cytometry. Sperm quality parameters (except counts) and VTG were significantly different among sites, with correlations between VTG and 7 sperm parameters. Thyrocyte heights, T4, T3, gonadosomatic index and Fulton's condition factor differed among sites, but not significantly. Sperm quality was significantly lower and VTG higher where liver contaminants and water estrogen equivalents were highest (LV site). Total PCBs (specifically PCB-138, -146, -151, -170, -174, -177, -180, -183, -187, -194, and -206) and total PBDEs (specifically BDE-47, -100, -153, and -154) were negatively correlated with sperm motility. PCB-206 and BDE-154 were positively correlated with DNA fragmentation, and pentachloroanisole and VTG were positively correlated with sperm apoptosis and negatively correlated with ATP. BDE-99 was positively correlated with sperm counts and motility; T4 was negatively correlated with counts and positively correlated with motility, thus indicating possible androgenic mechanisms and thyroid endocrine disruption. Male LSS proved to be an informative model for studying reproductive and endocrine biomarkers in the LCR.

Oregon;Washington↗

USGS2016

Hydrologic indicators of hot spots and hot moments of mercury methylation potential along river corridors

The biogeochemical cycling of metals and other contaminants in river-floodplain corridors is controlled by microbial activity responding to dynamic redox conditions. Riverine flooding thus has the potential to affect speciation of redox-sensitive metals such as mercury (Hg). Therefore, inundation history over a period of decades potentially holds information on past production of bioavailable Hg. We investigate this within a Northern California river system with a legacy of landscape-scale 19th century hydraulic gold mining. We combine hydraulic modeling, Hg measurements in sediment and biota, and first-order calculations of mercury transformation to assess the potential role of river floodplains in producing monomethylmercury (MMHg), a neurotoxin which accumulates in local and migratory food webs. We identify frequently inundated floodplain areas, as well as floodplain areas inundated for long periods. We quantify the probability of MMHg production potential (MPP) associated with hydrology in each sector of the river system as a function of the spatial patterns of overbank inundation and drainage, which affect long-term redox history of contaminated sediments. Our findings identify river floodplains as periodic, temporary, yet potentially important, loci of biogeochemical transformation in which contaminants may undergo change during limited periods of the hydrologic record. We suggest that inundation is an important driver of MPP in river corridors and that the entire flow history must be analyzed retrospectively in terms of inundation magnitude and frequency in order to accurately assess biogeochemical risks, rather than merely highlighting the largest floods or low-flow periods. MMHg bioaccumulation within the aquatic food web in this system may pose a major risk to humans and waterfowl that eat migratory salmonids, which are being encouraged to come up these rivers to spawn. There is a long-term pattern of MPP under the current flow regime that is likely to be accentuated by increasingly common large floods with extended duration.

California↗

USGS2018

Juvenile coho salmon growth and health in streams across an urbanization gradient

Expanding human population and urbanization alters freshwater systems through structural changes to habitat, temperature effects from increased runoff and reduced canopy cover, altered flows, and increased toxicants. Current stream assessments stop short of measuring health or condition of species utilizing these freshwater habitats and fail to link specific stressors mechanistically to the health of organisms in the stream. Juvenile fish growth integrates both external and internal conditions providing a useful indicator of habitat quality and ecosystem health. Thus, there is a need to account for ecological and environmental influences on fish growth accurately. Bioenergetics models can simulate changes in growth and consumption in response to environmental conditions and food availability to account for interactions between an organism's environmental experience and utilization of available resources. The bioenergetics approach accounts for how thermal regime, food supply, and food quality affect fish growth. This study used a bioenergetics modeling approach to evaluate the environmental factors influencing juvenile coho salmon growth among ten Pacific Northwest streams spanning an urban gradient. Urban streams tended to be warmer, have earlier emergence dates and stronger early season growth. However, fish in urban streams experienced increased stress through lower growth efficiencies, especially later in the summer as temperatures warmed, with as much as a 16.6% reduction when compared to fish from other streams. Bioenergetics modeling successfully characterized salmonid growth in small perennial streams as part of a more extensive monitoring program and provides a powerful assessment tool for characterizing mixed life-stage specific responses in urban streams.

Science of the Total Environment↗

USGS2017

A effective DNA vaccine against diverse genotype J infectious hematopoietic necrosis virus strains prevalent in China

Infectious hematopoietic necrosis virus (IHNV) is the most important pathogen threatening the aquaculture of salmonid fish in China. In this study, a DNA vaccine, designated pIHNch-G, was constructed with the glycoprotein (G) gene of a Chinese IHNV isolate SD-12 (also called Sn1203) of genotype J. The minimal dose of vaccine required, the expression of the Mx-1 gene in the muscle (vaccine delivery site) and anterior kidney, and the titers of the neutralizing antibodies produced were used to evaluate the vaccine efficacy. To assess the potential utility of the vaccine in controlling IHNV throughout China, the cross protective efficacy of the vaccine was determined by challenging fish with a broad range of IHNV strains from different geographic locations in China. A single 100 ng dose of the vaccine conferred almost full protection to rainbow trout fry (3 g) against waterborne or intraperitoneal injection challenge with IHNV strain SD-12 as early as 4 days post-vaccination (d.p.v.), and significant protection was still observed at 180 d.p.v. Intragenogroup challenges showed that the DNA vaccine provided similar protection to the fish against all the Chinese IHNV isolates tested, suggesting that the vaccine can be widely used in China. Mx-1 gene expression was significantly upregulated in the muscle tissue (vaccine delivery site) and anterior kidney in the vaccinated rainbow trout at both 4 and 7 d.p.v. Similar levels of neutralizing antibodies were determined with each of the Chinese IHNV strains at 60 and 180 d.p.v. This DNA vaccine should play an important role in the control of IHN in China.

Vaccine↗

USGS2011

Environmental endocrinology of salmon smoltification

Smolting is a hormone-driven developmental process that is adaptive for downstream migration and ocean survival and growth in anadromous salmonids . Smolting includes increased salinity tolerance, increased metabolism, downstream migratory and schooling behavior, silvering and darkened fin margins, and olfactory imprinting. These changes are promoted by growth hormone, insulin-like growth factor I, cortisol , thyroid hormones, whereas prolactin is inhibitory. Photoperiod and temperature are critical environmental cues for smolt development, and their relative importance will be critical in determining responses to future climate change. Most of our knowledge of the environmental control and endocrine mediation of smolting is based on laboratory and hatchery studies, yet there is emerging information on fish in the wild that indicates substantial differences. Such differences may arise from differences in environmental stimuli in artificial rearing environments, and may be critical to ocean survival and population sustainability. Endocrine disruptors, acidification and other contaminants can perturb smolt development, resulting in poor survival after seawater entry.

General and Comparative Endocrinology↗

USGS2021

Effects of long-term cortisol treatment on growth and osmoregulation of Atlantic salmon and brook trout

Cortisol is the final product of the hypothalamic-pituitary-interrenal (HPI) axis and acts as a gluco- and mineralo-corticoid in fish. Long-term elevations of cortisol have been linked to reduced growth in fishes, but the mechanism(s) and relative sensitivities of species are still unclear. We carried out experiments to examine the relative effects of cortisol on growth and gill NKA activity in two salmonids: Atlantic salmon ( Salmo salar ) and brook trout ( Salvelinus fontinalis ). Treatment with intraperitoneal cortisol implants for 30 days resulted in reduced growth in both species, but with greater sensitivity to cortisol in brook trout. Gill NKA activity was strongly upregulated by cortisol in Atlantic salmon, and weakly upregulated in brook trout but with no statistically significant effect. Cortisol treatment resulted in reduced plasma levels of insulin-like growth factor I and increased plasma growth hormone levels in Atlantic salmon. Our results demonstrate that there are species differences in the sensitivity of growth and osmoregulation to cortisol, even among species in the same family (Salmonidae).

General and Comparative Endocrinology↗

USGS2019

Phylogeography and evolution of infectious hematopoietic necrosis virus in China

Infectious hematopoietic necrosis virus (IHNV) is a well-known rhabdoviral pathogen of salmonid fish. In this study, a comprehensive analysis of 40 IHNV viruses isolated from thirteen fish farms in nine geographically dispersed Chinese provinces during 2012 to 2017 is presented. Identity of nucleotide and amino acid sequences among all the complete glycoprotein (G) genes from Chinese isolates was 98.0–100% and 96.7–100%, respectively. Coalescent phylogenetic analyses revealed that all the Chinese IHN virus characterized in this study were in a monophyletic clade that had a most recent common ancestor with the J Nagano (JN) subgroup within the J genogroup of IHNV. Within the Chinese IHNV clade isolates obtained over successive years from the same salmon fish farm clustered in strongly supported subclades, suggesting maintenance and diversification of virus over time within individual farms. There was also evidence for regional virus transmission within provinces, and some cases of longer distance transmission between distant provinces, such as Gansu and Yunnan. The data demonstrated that IHNV has evolved into a new subgroup in salmon farm environments in China, and IHNV isolates are undergoing molecular evolution within fish farms. We suggest that Chinese IHNV comprises a separate JC subgroup within the J genogroup of IHNV.

Molecular Phylogenetics and Evolution↗

USGS2001

Mucous lysozyme levels in hatchery coho salmon (Oncorhynchus kisutch) and spring chinook salmon (O. tshawytscha) early in the parr-smolt transformation

Mucous lysozyme concentrations were determined in juvenile coho salmon (Oncorhynchus kisutch) and spring chinook salmon (O. tshawytscha) to establish reference levels during the time associated with the parr-smolt transformation. The first reported naris and vent mucous lysozyme levels are provided for spring chinook salmon and coho salmon. Naris mucous lysozyme levels ranged between 300 and 700 ??g ml-1, vent mucous lysozyme from 100 to 300 ??g ml-1, and skin mucous lysozyme levels were below 130 ??g ml-1. Lysozyme levels in the two species showed the same relationship with the highest levels in naris mucous, and the lowest in skin mucous. A seasonal decrease occurred in both species with a significant decrease in naris mucous lysozyme between February and March. Gill ATPase levels used to monitor smolt development during the same period did not reach ranges reported for smolts for either species during emigration. Identification of seasonal levels of lysozyme activity in mucous provides an alternative determination of developmental status prior to release of fish from the hatchery when salmonids are still undergoing the parr-smolt transformation. ?? 2001 Elsevier Science B.V.

Aquaculture↗

USGS2003

Residue depletion of oxytetracycline from fillet tissues of northern pike and walleye

The broad-spectrum antibacterial drug oxytetracycline (OTC) is used in the U.S. to treat certain diseases in salmonids and catfish. This study was conducted to support an extension of the OTC label to include all cool-water fish species cultured at U.S. public aquaculture facilities by satisfying human food safety requirements. Juvenile northern pike ( Esox lucius ; mean weight: 117 g) and walleye ( Stizostedion vitreum ; mean weight: 59 g) were fed OTC-medicated diets near the maximum legal treatment rate (82.7 mg OTC-HCl/kg fish/day for 10 days) and near the lower limit of the water temperature range for most disease outbreaks in these species (14 and 16 °C, respectively). Two trials were conducted simultaneously with northern pike, one using commercially medicated feed and the other using on-site OTC top-coated feed. A third trial was performed with walleye using on-site OTC top-coated feed. Fillet tissues were collected and OTC free base (OTC-base) concentrations were determined by high performance liquid chromatography. The maximum mean OTC-base concentrations in the fillet tissue were 319 ng/g in northern pike (skinless) and 721 ng/g in walleye (skin-on), both well below the current tolerance limit of 2000 ng/g OTC-base. The log-linear loss of OTC-base from the fillet tissues was monophasic, with terminal phase half-lives of 5.9 days in northern pike fed commercial medicated feed, 6.7 days in northern pike fed top-coated feed, and 10.5 days in walleye fed top-coated feed. The data supported a zero withdrawal time in juvenile northern pike and walleye fed OTC at the approved dose level for 10 days at water temperatures down to 14 and 16 °C respectively.

Aquaculture↗

USGS1997

Safety of formalin treatments on warm- and coolwater fish eggs

Formalin is widely used for treating fungal infections of fish eggs in intensive aquaculture operations. The use of formalin in the United States is only allowed on salmonid and esocid eggs unless a special exemption is granted for use on other species. This study was conducted to determine the safety of formalin treatments on eggs of representative warm- and coolwater fish species and data was used to support a request to allow the use of formalin on the eggs of warmwater and additional coolwater fish species. Non-eyed eggs of walleye ( Stizostedion vitreum ), common carp ( Cyprinus carpio ), white sucker ( Catostomus commersoni ), channel catfish ( Ictalurus punctatus ), and lake sturgeon ( Acipenser fulvescens ) were cultured in miniature egg hatching jars and treated for 45 min every-other-day with 1500, 4500, or 7500 μL L -1 formalin up to hatch. For all species tested, the percent hatch was greater in 1500 mu L L-1 treatment groups than in untreated controls. Walleye eggs were the least sensitive species and had a hatch of 87% in the 7500 mu L L-1 treatment. Lake sturgeon were the most sensitive species with a mean hatch of 54% in 1500 mu L L-1 treatments. Adequate margins of safety exist for standard treatments (1500 mu L L-1 for 15 min) on eggs of all species tested except lake sturgeon. Fungal infections drastically reduced or eliminated hatch in most control groups whereas most treated groups were free of infections. This confirms the efficacy of formalin as an fungicide. Published by Elsevier Science B.V.

Aquaculture↗

USGS1985

Introduction and spread of the threespine stickleback ( Gasterosteus aculeatus ) in Lakes Huron and Michigan

The threespine stickleback ( Gasterosteus aculeatus ) was not known to occur in the Great Lakes above Niagara Falls until 1980, when it was collected in South Bay, Manitoulin Island, in the Lake Huron basin. By 1984 this species had been found in tributaries of Lakes Huron and Michigan, and in the open waters of both lakes. All specimens identified were the completely plated morph that is most prevalent in fresh water along the east coast of North America. The status of this species in Lakes Huron and Michigan appears to be “Possibly Established.” If threespine stickleback increase in abundance they may eventually provide additional forage for large salmonids.

Journal of Great Lakes Research↗

USGS2024

Structurally selective ozonolysis of p-phenylenediamines and toxicity in coho salmon and rainbow trout

The tire-rubber-derived ozonation product of N -(1,3-dimethylbutyl)- N ′-phenyl- p -phenylenediamine (6PPD), N -(1,3-dimethylbutyl)- N ′-phenyl- p -phenylenediamine-quinone (6PPD-Q), was recently discovered to cause acute mortality in coho salmon ( Oncorhynchus kisutch ). para -Phenylenediamines (PPDs) with variable side chains distinct from 6PPD have been identified as potential replacement antioxidants, but their toxicities remain unclear under environmentally relevant ozone conditions. We herein tested the multiphase gas-surface ozone reactivity of four select PPDs [6PPD, N -isopropyl- N ′-phenyl- p -phenylenediamine (IPPD), N , N ′-diphenyl- p -phenylenediamine (DPPD), and N -phenyl- N ′-cyclohexyl- p -phenylenediamine (CPPD)] and evaluated the toxicity of their reaction mixtures in coho salmon, rainbow trout ( Oncorhynchus mykiss ), and fathead minnow ( Pimephales promelas ). 6PPD and IPPD were found to rapidly react with ozone, while no significant multiphase ozone reactivity was observed for DPPD or CPPD. The viability of coho salmon CSE-119 cells was strongly affected by the ozonolysis products of 6PPD but not by those of the other three PPDs. Acute mortality was only observed in juvenile rainbow trout that were exposed to oxidized 6PPD, suggesting a common mechanism of toxic action in the two salmonid fish species. This study reports the structurally selective ozone reactivity of PPDs and the unique toxicity of 6PPD ozonolysis mixtures, which demonstrates that other PPDs are potential alternative antioxidants.

Environmental Science and Technology↗

USGS2023

Establishing an in vitro model to assess the toxicity of 6PPD-quinone and other tire wear transformation products

The tire wear transformation product 6PPD-quinone (6PPDQ) has been implicated as the causative factor for broad scale mortality events for coho salmon in the Pacific Northwest. Highly variable sensitivity to 6PPDQ in closely related salmonids complicates efforts to evaluate the broader toxicological impacts to aquatic ecosystems. Our goals were to (1) validate the large range of in vivo species sensitivities reported for coho, Chinook, and sockeye salmon and (2) develop an in vitro platform for assessing 6PPDQ toxicity. In vivo studies confirmed the acute sensitivity of juvenile coho (12 h LC 50 = 80.4 ng/L) and demonstrated that sockeye salmon were not vulnerable to mortality. Chinook salmon were sensitive to 6PPDQ mortality at initial concentrations >25 μg/L, ∼10-fold greater than reported environmental measurements. In vitro , the coho salmon cell line CSE-119 was acutely sensitive to 6PPDQ (metabolic EC 50 = 7.9 μg/L, cytotoxicity EC 50 = 6.1 μg/L). Analogous Chinook (CHSE-214) and sockeye salmon (SSE-5) cell lines were nonresponsive in both assays, and rainbow trout RTG-2 cells began showing metabolic effects at 68 μg/L (EC 5 ). Recreation of species-specific 6PPDQ sensitivity in vitro implicates conserved modes of action in CSE-119 that could be utilized for mechanistic studies of 6PPDQ toxicity and screening of other PPD transformation products.

Environmental Science and Technology Letters↗

USGS2005

Speciation of mercury and mode of transport from placer gold mine tailings

Historic placer gold mining in the Clear Creek tributary to the Sacramento River (Redding, CA) has highly impacted the hydrology and ecology of an important salmonid spawning stream. Restoration of the watershed utilized dredge tailings contaminated with mercury (Hg) introduced during gold mining, posing the possibility of persistent Hg release to the surrounding environment, including the San Francisco Bay Delta. Column experiments have been performed to evaluate the extent of Hg transport under chemical conditions potentially similar to those in river restoration projects utilizing dredge tailings such as at Clear Creek. Physicochemical perturbations, in the form of shifts in column influent ionic strength and the presence of a low molecular weight organic acid, were applied to coarse and fine sand placer tailings containing 109-194 and 69-90 ng of Hg/g, respectively. Significant concentrations of mercury, up to 16 ??g/L, leach from these sediments in dissolved and particle-associated forms. Sequential chemical extractions (SCE) of these tailings indicate that elemental Hg initially introduced during gold mining has been transformed to readily soluble species, such as mercury oxides and chlorides (3-4%), intermediately extractable phases that likely include (in)organic sorption complexes and amalgams (75-87%), and fractions of highly insoluble forms such as mercury sulfides (6-20%; e.g., cinnabar and metacinnabar). Extended X-ray absorption fine structure (EXAFS) spectroscopic analysis of colloids obtained from column effluent identified cinnabar particles as the dominant mobile mercury-bearing phase. The fraction of intermediately extractable Hg phases also likely includes mobile colloids to which Hg is adsorbed. ?? 2005 American Chemical Society.

Environmental Science & Technology↗

USGS2001

Copper, cadmium, and zinc concentrations in juvenile Chinook salmon and selected fish-forage organisms (aquatic insects) in the upper Sacramento River, California

This study assessed the downstream extent andseverity of copper (Cu), cadmium (Cd), and zinc (Zn)contamination from acid mine drainage on juvenile chinook salmon( Oncorhynchus tshawytscha ) and aquatic insects over aroughly 270-km reach of the Sacramento River below KeswickReservoir. During April–May 1998, salmon were collected fromfour sites in the river and from a fish hatchery that receiveswater from Battle Creek. Salmon from river sites were examinedfor gut contents to document their consumption of variousinvertebrate taxa, whereas salmon from river sites and thehatchery were used for metal determinations. Midge(Chironomidae) and caddisfly (Trichoptera) larvae and mayfly(Ephemeroptera) nymphs were collected for metal determinationsduring April–June from river sites and from Battle and Buttecreeks. The fish hatchery and Battle and Butte creeks served asreference sites because they had no history of receiving minedrainage. Salmon consumed mostly midge larvae and pupae (44.0%,damp-dry biomass), caddisfly larvae (18.9%), Cladocera (5.8%),and mayfly nymphs (5.7%). These results demonstrated thatinsects selected for metal determinations were important as fishforage. Dry-weight concentrations of Cu, Cd, and Zn weregenerally far higher in salmon and insects from the river thanfrom reference sites. Within the river, high metalconcentrations persisted as far downstream as South Meridian (thelowermost sampling site). Maximum concentrations of Cd (30.7 μg g -1 ) and Zn (1230 μg g -1 ),but not Cu (87.4 μg g -1 ), in insects exceeded amounts that other investigators reported as toxic when fed for prolonged periods to juvenile salmonids.

California↗

USGS2002

Inhibition of erythrocytes δ-aminolevulinic acid dehydratase (ALAD) activity in fish from waters affected by lead smelters

We assessed the effects on fish of lead (Pb) released to streamsby smelters located in Trail, BC (Canada), E. Helena, MT, Herculaneum, MO, and Glover, MO. Fish were collected by electrofishing from sites located downstream of smelters and from reference sites. Blood from each fish was analyzed for δ-aminolevulinic acid dehydratase ( ALAD ) activity and hemoglobin (Hb), and samples of blood, liver, or carcass were analyzed for Pb, zinc (Zn), or both. Fish collected downstreamof all four smelters sites had elevated Pb concentrations, decreased ALAD activity, or both relative to their respectivereference sites. At E. Helena, fish from the downstream site also had lower Hb concentrations than fish from upstream. Differences among taxa were also apparent. Consistent with previous studies, ALAD activity in catostomids ( Pisces : Catostomidae-northern hog sucker, Hypentelium nigricans ;river carpsucker, Carpiodes carpio ; largescale sucker, Catostomus macrocheilus ; and mountain sucker, C. platyrhynchus ) seemed more sensitive to Pb-induced ALAD inhibition than the salmonids ( Pisces : Salmonidae-rainbow trout, Oncorhynchus mykiss ; brook trout, Salvelinus fontinalis ) or common carp ( Cyprinus carpio ). Some of these differences may have resulted from differential accumulation of Zn, which was not measured at all sites. We detected no ALAD activity in channel catfish ( Ictaluruspunctatus ) from either site on the Mississippi River at Herculaneum, MO. Our findings confirmed that Pb is releasedto aquatic ecosystems by smelters and accumulated by fish, andwe documented potentially adverse effects of Pb in fish. We recommend that Zn be measured along with Pb when ALAD activityis used as a biomarker and the collection of at least 10 fish ofa species at each site to facilitate statistical analysis.

Environmental Monitoring and Assessment↗

USGS2004

Major histocompatibility complex loci are associated with susceptibility of Atlantic salmon to infectious hematopoietic necrosis virus

Infectious hematopoietic necrosis virus (IHNV) is one of the most significant viral pathogens of salmonids and is a leading cause of death among cultured juvenile fish. Although several vaccine strategies have been developed, some of which are highly protective, the delivery systems are still too costly for general use by the aquaculture industry. More cost effective methods could come from the identification of genes associated with IHNV resistance for use in selective breeding. Further, identification of susceptibility genes may lead to an improved understanding of viral pathogenesis and may therefore aid in the development of preventive and therapeutic measures. Genes of the major histocompatibility complex (MHC), involved in the primary recognition of foreign pathogens in the acquired immune response, are associated with resistance to a variety of diseases in vertebrate organisms. We conducted a preliminary analysis of MHC disease association in which an aquaculture strain of Atlantic salmon was challenged with IHNV at three different doses and individual fish were genotyped at three MHC loci using denaturing gradient gel electrophoresis (PCR-DGGE), followed by sequencing of all differentiated alleles. Nine to fourteen alleles per exon-locus were resolved, and alleles potentially associated with resistance or susceptibility were identified. One allele ( Sasa -B-04) from a potentially non-classical class I locus was highly associated with resistance to infectious hematopoietic necrosis ( p < 0.01). This information can be used to design crosses of specific haplotypes for family analysis of disease associations.

Environmental Biology of Fishes↗