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USGS1994

Neutralization-resistant variants of infectious hematopoietic necrosis virus have altered virulence and tissue tropism

Infectious hematopoietic necrosis virus (IHNV) is a rhabdovirus that causes an acute disease in salmon and trout. In this study, a correlation between changes in tissue tropism and specific changes in the virus genome appeared to be made by examining four IHNV neutralization-resistant variants (RB-1, RB-2, RB-3, and RB-4) that had been selected with the glycoprotein (G)-specific monoclonal antibody RB/B5. These variants were compared with the parental strain (RB-76) for their virulence and pathogenicity in rainbow trout after waterborne challenge. Variants RB-2, RB-3, and RB-4 were only slightly attenuated and showed distributions of viral antigen in the livers and hematopoietic tissues of infected fish similar to those of the parental strain. Variant RB-1, however, was highly attenuated and the tissue distribution of viral antigen in RB-1-infected fish was markedly different, with more viral antigen in brain tissue. The sequences of the G genes of all four variants and RB-76 were determined. No significant changes were found for the slightly attenuated variants, but RB-1 G had two changes at amino acids 78 and 218 that dramatically altered its predicted secondary structure. These changes are thought to be responsible for the altered tissue tropism of the virus. Thus, IHNV G, like that of rabies virus and vesicular stomatitis virus, plays an integral part in the pathogenesis of viral infection.

Journal of Virology↗

USGS2026

Disease-associated mortality drives reduction in Yukon River Chinook salmon escapement: A novel method for quantifying the negative impacts of ‘misfit’ parasites to improve fisheries management

Parasites can suppress host populations through parasite-induced mortality. However, the negative effects of parasites are difficult to measure in wild populations and we have few tools for quantifying the magnitude of parasite-induced mortality. This is especially true for many ‘misfit’ parasites that do not fit into standard classifications (e.g. fungal-like parasites and myxozoans). As such, we are limited in our ability to include the effects of parasites in fish and wildlife management strategies. Chinook salmon ( Oncorhynchus tshawytscha ) are a species of immense cultural and ecological significance but populations are declining across much of their range. In the Yukon River, Alaska, USA, episodic outbreaks of the ichthyosporean parasite Ichthyophonus have been linked to declines, but population-level effects of Ichthyophonus on Yukon Chinook salmon remain largely unknown. We developed a novel model that leverages changes in parasite intensity distributions to quantify the magnitude of disease-induced mortality occurring in host populations. We used it to address two questions to inform management of Yukon Chinook salmon: (i) Is parasite-induced mortality occurring in Chinook salmon and at what magnitude? (ii) What are the drivers of spatio-temporal variability in parasite-induced mortality? Using 3 years of surveys, we quantified the evidence for the presence, magnitude, and variability in Ichthyophonus -induced mortality in Chinook salmon. Our model predicted that Ichthyophonus was responsible for between 8% and 15% mortality of the migrating Chinook salmon population prior to reaching the Canadian border and could describe nearly 35% of unaccounted for mortality at the border. The model predicted that variability in mortality among years could largely be explained by differences in parasite acquisition in the marine environment rather than differences in infection dynamics in the river. Synthesis and applications . Alaska Department of Fish and Game is implementing an annual monitoring program at the mouth of the Yukon River where our model will estimate the proportion of fish at risk from parasite-induced mortality to inform annual management. Moreover, the model is broadly applicable to other fungal-like and myxozoan parasites of conservation concern, where improved estimates of parasite-induced mortality could be used to predict parasite suppression of wild and managed host populations.

Alaska↗

USGS2019

Performance of a proposed determinative method for p-TSA in rainbow trout fillet tissue and bridging the proposed method with a method for total chloramine-T residues in rainbow trout fillet tissue

Chloramine-T is an effective drug for controlling fish mortality caused by bacterial gill disease. As part of the data required for approval of chloramine-T use in aquaculture, depletion of the chloramine-T marker residue (para-toluenesulfonamide; p-TSA) from edible fillet tissue of fish must be characterized. Declaration of p-TSA as the marker residue for chloramine-T in rainbow trout was based on total residue depletion studies using a method that used time consuming and cumbersome techniques. A simple and robust method recently developed is being proposed as a determinative method for p-TSA in fish fillet tissue. The proposed determinative method was evaluated by comparing accuracy and precision data with U.S. Food and Drug Administration criteria and by bridging the method to the former method for chloramine-T residues. The method accuracy and precision fulfilled the criteria for determinative methods; accuracy was 92.6, 93.4, and 94.6% with samples fortified at 0.5X, 1X, and 2X the expected 1000 ng/g tolerance limit for p-TSA, respectively. Method precision with tissue containing incurred p-TSA at a nominal concentration of 1000 ng/g ranged from 0.80 to 8.4%. The proposed determinative method was successfully bridged with the former method. The concentrations of p-TSA developed with the proposed method were not statistically different at p < 0.05 from p-TSA concentrations developed with the former method.

Journal of AOAC International↗

USGS2020

Liquid chromatographic determination of para-toluenesulfonamide in edible fillet tissues from three species of fish

Chloramine-T ( N -sodium- N -chloro- p -toluene-sulfonamide) is a candidate therapeutic drug for treating bacterial gill disease, a predominant disease of a variety of fish species. Research has been initiated to obtain the U.S. Food and Drug Administration’s (FDA) approval for the use of chloramine-T on a variety of fish species. An attribute of a therapeutic aquaculture drug that must be characterized before the FDA approves its use is depletion of the drug’s marker residue (the drug’s parent compound or metabolite of highest concentration in an edible tissue). para -Toluenesulfonamide (p-TSA) is the primary degradation product and marker residue for chloramine-T in rainbow trout. To conduct residue depletion studies for chloramine-T in fish, a robust analytical method sensitive and specific for p-TSA residues in edible fillet tissue from a variety of fish was required. Homogenized fillet tissues from rainbow trout ( Oncorhynchus mykiss ), walleye ( Stizostedion vitreum ), and channel cattish ( Ictalurus punctatus ) were fortified at nominal p-TSA concentrations of 17, 67, 200, 333, and 1000 ng/g. Samples were analyzed by isocratic reversed-phase liquid chromatography (LC) with absorbance detection at 226 nm. Mean recoveries of p-TSA ranged from 77 to 93.17%; relative standard deviations ranged from 1.5 to 14%; method quantitation limits ranged from 13 to 18 ng/g; and method detection limits ranged from 3.8 to 5.2 ng/g. The LC parameters produced p-TSA peaks without coelution of endogenous compounds and excluded chromatographic interference from at least 20 chemicals and drugs of potential use in aquaculture.

Journal of AOAC International↗

USGS2011

The effects of myxobolus cerebralis on the physiological performance of whirling disease resistant and susceptible strains of rainbow trout

The development of rainbow trout Oncorhynchus mykiss strains that are resistant to whirling disease has shown promise as amanagement tool for populationsin areas where Myxobolus cerebralisis present. However, the physiological effects of the disease on characteristics necessary for fish survival in natural river conditions have not been tested in many of these strains. Five rainbow trout strains were evaluated for their swimming ability and growth characteristics in relation to M. cerebralis exposure: the resistant German rainbow trout (GR) strain (Hofer strain), the susceptible Colorado River rainbow trout (CRR) strain, and three intermediate (hybrid) strains (F1 = GR ?? CRR; F2 = F1 ?? F1; B2 = backcross of F1 ?? CRR). Three broad response patterns among strain and exposure were evident in our study. First, exposure metrics, growth performance, and swimming ability differed among strains. Second, exposure to the parasite did not necessarily produce differences in growth or swimming ability. Exposure to M. cerebralis did not affect batch weight for any strain, and critical swimming velocity did not differ between exposed and unexposed families. Third, although exposure did not necessarily affect growth or swimming ability, individuals that exhibited clinical deformities did show reduced growth and swimming performance; fish with clinical deformities were significantly smaller and had lower critical swimming velocities than exposed fish without clinical deformities. Research and management have focused on GR ?? CRR hybrid strains; however, given the performance of the GR strain in our study, it should not be discounted as a potential broodstock. Additional field trials comparing the GR and F1 strains should be conducted before wholesale adoption of the GR strain to reestablish rainbow trout populations in Colorado. ?? American Fisheries Society 2011.

Journal of Aquatic Animal Health↗

USGS2018

Survival of whirling disease resistant rainbow trout fry in the wild: A comparison of two strains

Introduced pathogens can affect fish populations, and three main factors affect disease occurrence: the environment, host, and pathogen. Manipulating at least one of these factors is necessary for controlling disease. Myxobolus cerebralis , the parasite responsible for salmonid whirling disease, became established in Colorado during the 1990s and caused significant declines in wild Rainbow Trout Oncorhynchus mykiss populations. Attempts to re-establish Rainbow Trout have focused on manipulating salmonid host resistance. A Rainbow Trout strain known as GR × CRR was developed for stocking in Colorado by crossing a whirling-disease-resistant strain known as the German Rainbow Trout (GR) with the Colorado River Rainbow Trout (CRR). The GR × CRR fish exhibit resistance similar to that shown by GR, and survival and reproduction were expected to be similar to those of CRR. One disadvantage of stocking GR × CRR is that outcrossing and backcrossing could decrease resistance, and laboratory studies have indicated that this can occur. A potential disadvantage of stocking pure GR is lower survival due to domestication. To compare fry survival between the strains, a field experiment was conducted in 1.6-km reaches of nine Colorado streams. Each stream was stocked in August 2014 with 5,000 GR × CRR and 5,000 GR individuals. In October 2014, April 2015, and August 2015, apparent survival was assessed. Two laboratory predation experiments were also conducted. The field experiment revealed that short-term apparent survival was influenced by stream, and growth rate was influenced by strain and stream. However, after 12 months, there was no difference in apparent survival or growth rate between the GR and GR × CRR strains. Laboratory experiments showed that survival did not differ between the strains when confronted with Brown Trout Salmo trutta predation. Our results indicate that the GR strain is a viable option for stocking in streams where M. cerebralis is enzootic. Further evaluation is needed to determine whether GR fish will survive to maturity and reproduce.

Journal of Aquatic Animal Health↗

USGS2009

Propagation of the myxozoan parasite Myxobolus cerebralis by different geographic and genetic populations of Tubifex tubifex: An Oregon perspective

Tubifex tubifex are obligate invertebrate hosts in the life cycle of Myxobolus cerebralis, the myxozoan parasite that causes whirling disease in salmonid fishes. This exotic parasite is established to varying degrees across Oregon's Columbia River system (Pacific Northwest, USA) and characteristics of local T. tubifex populations likely play a role in the pattern of disease occurrence. To better understand these patterns, we collected T. tubifex from three Oregon river basins (Willamette, Deschutes, and Grande Ronde), determined their genotype (mitochondrial 16S rDNA lineage and RAPD genotype) and exposed 10 different populations to M. cerebralis in the laboratory. Four mt lineages were identified: I, III, V and VI. Lineage III was found in all river basins but dominated both central and eastern sites. The RAPD assay further divided these lineages into geographic sub-populations; no RAPD genotype was common to all basins. There was a significant difference in prevalence of infection and level of parasite production among the populations we exposed to M. cerebralis that was attributed to genotypic composition. Only lineage III worms released actinospores and only populations dominated by this lineage amplified the parasite. These populations had the lowest survival, however, the lineage dominant before exposure remained dominant despite the high prevalence of infection. The distribution and infection dynamics of susceptible T. tubifex throughout Oregon may contribute to the differences in M. cerebralis occurrence; our studies further support the influence of oligochaete genotypes on the manifestation of whirling disease in salmonid populations. ?? 2009 Elsevier Inc. All rights reserved.

Journal of Invertebrate Pathology↗

USGS2001

Evidence that coded-wire-tagging procedures can enhance transmission of Renibacterium salmoninarum in chinook salmon

Binary coded wire tags (CWTs) are used extensively for identification and management of anadromous salmonid populations. A study of bacterial kidney disease (BKD) in two brood year groups of hatchery-reared spring chinook salmon Oncorhynchus tshawytscha provided strong evidence that horizontal transmission of Renibacterium salmoninarum, the causative agent of BKD, might be enhanced by CWT-marking procedures. About 4 months after CWTs were implanted in the snouts of juvenile fish, 14-16 different tissues were sampled from each of 60 fish per brood year group for histological analysis. Of the fish that were positive for R. salmoninarum by histological examination, 41% (7 of 17) of the 1988 brood year fish and 24% (10 of 42) of the 1989 brood year fish had BKD lesions confined to the head near the site of tag implantation. These lesions often resulted in the destruction of tissues of one or both olfactory organs. No focal snout infections were observed in fish that had not been marked with CWTs. Further data obtained from tissue analyses by use of an enzyme-linked immunosorbent assay and a fluorescent antibody test for detection of R. salmoninarum supported the hypothesis that infections of R. salmoninarum can be initiated in the snout tissues of CWT-marked fish and then spread to other organs. The tagging procedures might promote transmission of the pathogen among fish via contaminated tagging needles, by facilitating the entry of pathogens through the injection wound, or both. Limited evidence from this study suggested that implantation of passive integrated transponder tags in the peritoneal cavities of fish might also promote the transmission of R. salmoninarum or exacerbate existing infections. The results indicated a need for strict sanitary procedures during the tagging of fish in populations positive for R. salmoninarum to reduce the probability of enhanced horizontal transmission of the pathogen.

Journal of Aquatic Animal Health↗

USGS2021

Effects of stocking density on stress response and susceptibility to infectious hematopoietic necrosis virus in rainbow trout

The goals of this study were to examine the effect of stocking density on the stress response and disease susceptibility in juvenile rainbow trout ( Oncorhynchus mykiss ). Fish were sorted into one of 2 stocking densities (high density "HD", 20-40 kg/m 3 ) or (low density, "LD", 4-8 kg/m 3 ) and 3 stress indices (cortisol levels in serum and water, and neutrophil: lymphocyte (N:L) ratios from blood smears) were measured at multiple time points over 21 d. Serum cortisol was significantly increased at 1 h in LD samples and at 14 d in HD samples. Water cortisol concentrations were significantly higher in LD tanks as compared with HD tanks on day 14. N:L ratios were significantly higher in HD tanks on day 14 as compared with LD tanks and with baseline. The effect of stocking density on mortality after exposure to infectious hematopoietic necrosis virus (IHNV) was compared between fish held in HD or LD conditions, with or without prior acclimation to the different density conditions. No significant differences in survival were found between HD and LD treatments or between acclimated and nonacclimated treatments. Cumulative results indicate that 1) 1 to 4 gram rainbow trout did not generally demonstrate significant differences in stress indices at the density conditions tested over a 21-d period, 2) independent differences were found in 3 stress indices at day 14 after sorting into LD and HD holding conditions; and 3) LD and HD stocking densities did not have a significant effect on mortality due to IHNV.

Journal of the American Association for Laboratory↗

USGS2014

Effectiveness of Aquaflor (50% florfenicol) administered in feed to control mortality associated with Streptococcus iniae in tilapia at a commercial tilapia production facility

The efficacy of Aquaflor (florfenicol; FFC) to control mortality caused by Streptococcus iniae in tilapia was evaluated under field conditions. The trial was initiated following presumptive diagnosis of S. iniae infection in a mixed group of fingerling (mean, 4.5 g) Nile Tilapia Oreochromis niloticus and a hybrid of Nile Tilapia×Blue Tilapia O. aureus . Diagnoses included mortality in source tank; examination of clinical signs and presence or absence of gram-positive cocci in brain, and collection of samples for microbiological review and disease confirmation of 60 moribund fish. Following presumptive diagnosis, tilapia (83/tank) were randomly transferred to each of 20 test tanks receiving the same water as the source tank (test tank water was not reused). Tilapia were offered either nonmedicated control feed or FFC-medicated feed (FFC at 15 mg/kg body weight/d; 10 tanks per regimen) for 10 consecutive days followed by a 14-d observation period during which only the nonmedicated control feed was offered. Streptococcus iniae was presumptively identified during pretreatment necropsy and confirmed by polymerase chain reaction assay; S. iniae was confirmed in samples taken during the dosing period but was not detected during the postdosing period. The FFC disk diffusion zone of inhibition ranged from 29 to 32 mm, while the minimum inhibitory concentration of FFC ranged from 2 to 4 μg/mL for the S. iniae isolates collected. Survival of tilapia assigned to the FFC-dose group was significantly greater at 14 d posttreatment than that of the nonmedicated controls. The odds of tilapia assigned to the FFC-dose group surviving to the end of the postdosing period were 1.34 times the odds of survival of tilapia assigned to the nonmedicated control group. There were no clinically apparent adverse effects associated with the administration of FFC-medicated feed in this study.

North American Journal of Aquaculture↗

USGS1998

Molecular approaches to fish vaccines

For more than 50 years, researchers have tested a variety of killed, attenuated, and subunit preparations for control offish diseases. The earliest fish vaccines used killed preparations containing whole bacteria, viruses, or parasites and today, several bacterins have become commercially successful with more expected as improved delivery systems and adjuvants are realized. Live, attenuated vaccines have been developed by serial passage of a pathogen in culture or by using naturally occurring mutants and cross-reacting strains. These generally offer excellent protection and are cost-effective, but concerns about residual virulence or their effects on other aquatic species make them difficult candidates for licensing. In recent years, the tools of molecular biology have been applied to construction of a variety of recombinant, engineered, or subunit vaccines for fish. Among the approaches to be discussed are: attenuated strains of viruses and bacteria created by deletion of specific genes associated with virulence, in vitro synthesis of protective antigens from genes cloned into E. coli or baculovirus expression systems, chemical synthesis of peptides that represent protective epitopes, and direct immunization with DNA coding for protective antigens. Preparations representing each of these approaches have been tested in laboratory or field trials with various results and such vaccines promise to be safe and relatively inexpensive if they are able to provide protection when delivered by immersion. A significant advantage of genetically engineered vaccines is the ability to construct multivalent preparations that can protect fish against several pathogens or different strains of the same pathogen. While many of these novel vaccine strategies have been effective at stimulating specific immunity in the laboratory, more work is needed to develop better delivery systems and to overcome potential regulatory concerns.

Journal of Applied Ichthyology↗

USGS2011

Occurrence of antibiotic resistance and characterization of resistant genes and integrons in Enterobacteriaceae isolated from integrated fish farms south China

Antibiotics are still widely applied in animal husbandry to prevent diseases and used as feed additives to promote animal growth. This could result in antibiotic resistance to bacteria and antibiotic residues in animals. In this paper, Enterobacteriaceae isolated from four integrated fish farms in Zhongshan, South China were tested for antibiotic resistance, tetracycline resistance genes, sulfonamide resistance genes, and class 1 integrons. The Kirby-Bauer disk diffusion method and polymerase chain reaction (PCR) assays were carried out to test antibiotic susceptibility and resistance genes, respectively. Relatively high antibiotic resistance frequencies were found, especially for ampicillin (80%), tetracycline (52%), and trimethoprim (50%). Out of 203 Enterobacteriaceae isolates, 98.5% were resistant to one or more antibiotics tested. Multiple antibiotic resistance (MAR) was found highest in animal manures with a MAR index of 0.56. Tetracycline resistance genes ( tet (A), tet (C)) and sulfonamide resistance genes ( sul2 ) were detected in more than 50% of the isolates. The intI1 gene was found in 170 isolates (83.7%). Both classic and non-classic class 1 integrons were found. Four genes, aadA5 , aadA22 , dfr2 , and dfrA17 , were detected. To our knowledge, this is the first report for molecular characterization of antibiotic resistance genes in Enterobacteriaceae isolated from integrated fish farms in China and the first time that gene cassette array dfrA17-aadA5 has been detected in such fish farms. Results of this study indicated that fish farms may be a reservoir of highly diverse and abundant antibiotic resistant genes and gene cassettes. Integrons may play a key role in multiple antibiotic resistances posing potential health risks to the general public and aquaculture.

Zhongshan↗

USGS2014

Florfenicol residues in Rainbow Trout after oral dosing in recirculating and flow-through culture systems

Aquaflor is a feed premix for fish containing the broad spectrum antibacterial agent florfenicol (FFC) incorporated at a ratio of 50% (w/w). To enhance the effectiveness of FFC for salmonids infected with certain isolates of Flavobacterium psychrophilum causing coldwater disease, the FFC dose must be increased from the standard 10 mg·kg −1 body weight (BW)·d −1 for 10 consecutive days. A residue depletion study was conducted to determine whether FFC residues remaining in the fillet tissue after treating fish at an increased dose would be safe for human consumption. Groups of Rainbow Trout Oncorhynchus mykiss (total n = 144; weight range, 126–617 g) were treated with FFC at 20 mg·kg −1 BW·d −1 for 10 d in a flow-through system (FTS) and a recirculating aquaculture system (RAS) each with a water temperature of ∼13°C. The two-tank RAS included a nontreated tank containing 77 fish. Fish were taken from each tank (treated tank, n = 16; nontreated tank, n = 8) at 6, 12, 24, 48, 72, 120, 240, 360, and 480 h posttreatment. Florfenicol amine (FFA) concentrations (the FFC marker residue) in skin-on fillets from treated fish were greatest at 12 h posttreatment (11.58 μg/g) in the RAS and were greatest at 6 h posttreatment (11.09 μg/g) in the FTS. The half-lives for FFA in skin-on fillets from the RAS and FTS were 20.3 and 19.7 h, respectively. Assimilation of FFC residues in the fillets of nontreated fish sharing the RAS with FFC-treated fish was minimal. Florfenicol water concentrations peaked in the RAS-treated tank and nontreated tanks at 10 h (453 μg/L) and 11 h (442 μg/L) posttreatment, respectively. Monitoring of nitrite concentrations throughout the study indicated the nitrogen oxidation efficiency of the RAS biofilter was minimally impacted by the FFC treatment.

Journal of Aquatic Animal Health↗

USGS2014

Contrasting past and current numbers of bears visiting Yellowstone cutthroat trout streams

Spawning cutthroat trout ( Oncorhynchus clarkii bouvieri ) were historically abundant within tributary streams of Yellowstone Lake within Yellowstone National Park and were a highly digestible source of energy and protein for Yellowstone’s grizzly bears ( Ursus arctos ) and black bears ( U. americanus ). The cutthroat trout population has subsequently declined since the introduction of non-native lake trout (Salvelinus namaycush), and in response to effects of drought and whirling disease ( Myxobolus cerebralis ). The trout population, duration of spawning runs, and indices of bear use of spawning streams had declined in some regions of the lake by 1997–2000. We initiated a 3-year study in 2007 to assess whether numbers of spawning fish, black bears, and grizzly bears within and alongside stream corridors had changed since 1997– 2000. We estimated numbers of grizzly bears and black bears by first compiling encounter histories of individual bears visiting 48 hair-snag sites along 35 historically fished streams.We analyzed DNA encounter histories with Pradel-recruitment and Jolly-Seber (POPAN) capture-mark-recapture models. When compared to 1997–2000, the current number of spawning cutthroat trout per stream and the number of streams with cutthroat trout has decreased. We estimated that 48 (95% CI¼42–56) male and 23 (95% CI¼21–27) female grizzly bears visited the historically fished tributary streams during our study. In any 1- year, 46 to 59 independent grizzly bears (8–10% of estimated Greater Yellowstone Ecosystem population) visited these streams. When compared with estimates from the 1997 to 2000 study and adjusted for equal effort, the number of grizzly bears using the stream corridors decreased by 63%. Additionally, the number of black bears decreased between 64% and 84%. We also document an increased proportion of bears of both species visiting front-country (i.e., near human development) streams. With the recovery of cutthroat trout, we suggest bears that still reside within the Lake basin will readily use this high-quality food resource.

Journal of Wildlife Management↗

USGS2016

Identification of the major capsid protein of erythrocytic necrosis virus (ENV) and development of quantitative real-time PCR assays for quantification of ENV DNA

Viral erythrocytic necrosis (VEN) is a disease of marine and anadromous fish that is caused by the erythrocytic necrosis virus (ENV), which was recently identified as a novel member of family Iridoviridae by next-generation sequencing. Phylogenetic analysis of the ENV DNA polymerase grouped ENV with other erythrocytic iridoviruses from snakes and lizards. In the present study, we identified the gene encoding the ENV major capsid protein (MCP) and developed a quantitative real-time PCR (qPCR) assay targeting this gene. Phylogenetic analysis of the MCP gene sequence supported the conclusion that ENV does not group with any of the currently described iridovirus genera. Because there is no information regarding genetic variation of the MCP gene across the reported host and geographic range for ENV, we also developed a second qPCR assay for a more conserved ATPase-like gene region. The MCP and ATPase qPCR assays demonstrated good analytical and diagnostic sensitivity and specificity based on samples from laboratory challenges of Pacific herring Clupea pallasii . The qPCR assays had similar diagnostic sensitivity and specificity as light microscopy of stained blood smears for the presence of intraerythrocytic inclusion bodies. However, the qPCR assays may detect viral DNA early in infection prior to the formation of inclusion bodies. Both qPCR assays appear suitable for viral surveillance or as a confirmatory test for ENV in Pacific herring from the Salish Sea.

Journal of Veterinary Diagnostic Investigation↗

USGS2005

Combined use of the ASK and SHK-1 cell lines to enhance the detection of infectious salmon anemia virus

Infectious salmon anemia (ISA) is a severe disease primarily affecting commercially farmed Atlantic salmon (Salmo salar) in seawater. The disease has been reported in portions of Canada, the United Kingdom, the Faroe Islands, and the United States. Infectious salmon anemia virus (ISAV), the causative agent of ISA, has also been isolated from several asymptomatic marine and salmonid fish species. Diagnostic assays for the detection of ISAV include virus isolation in cell culture, a reverse transcriptase-PCR, an enzyme-linked immunosorbent assay, and an indirect fluorescent antibody test. Virus isolation is considered the gold standard, and 5 salmonid cell lines are known to support growth of ISAV. In this study, the relative performance of the salmon head kidney 1 (SHK-1), Atlantic salmon kidney (ASK), and CHSE-214 cell lines in detecting ISAV was evaluated using samples from both experimentally and naturally infected Atlantic salmon. Interlaboratory comparisons were conducted using a quality control-quality assurance ring test. Both the ASK and SHK-1 cell lines performed well in detecting ISAV, although the SHK-1 line was more variable in its sensitivity to infection and somewhat slower in the appearance of cytopathic effect. Relative to the SHK-1 and ASK lines, the CHSE-214 cell line performed poorly. Although the ASK line appeared to represent a good alternative to the more commonly used SHK-1 line, use of a single cell line for diagnostic assays may increase the potential for false-negative results. Thus, the SHK-1 and ASK cell lines can be used in combination to provide enhanced ability to detect ISAV.

Journal of Veterinary Diagnostic Investigation↗

USGS2014

Surveillance and monitoring of white-tailed deer for chronic wasting disease in the northeastern United States

Chronic wasting disease (CWD) is a prion disease that affects both wild and captive cervid populations. In the past 45 y, CWD has spread from northern Colorado to all bordering states, as well as the midwestern United States (Midwest) and northeastern United States (Northeast), Canada, and South Korea. Because CWD is a relatively new issue for wildlife management agencies in the Northeast, we surveyed a representative (e.g., cervid biologist, wildlife veterinarian) from 14 states to gain a better understanding of state-specific surveillance measures. Between 2002 and 2012, New York (37,093) and Pennsylvania (35,324) tested the greatest number of harvested white-tailed deer Odocoileus virginianus in the Northeast. Additionally, the 14 states surveyed have tested 121,730 harvested deer, or approximately 15,216/y, since CWD was first detected in 2005. The most common tissues used by agencies in the Northeast for testing were retropharyngeal lymph nodes, which have been determined to be the most reliable in detecting CWD in cervids. Understanding CWD surveillance efforts at a regional scale can help to provide guidance for the development of new surveillance plans or the improvement of existing ones. Furthermore, collaborations among state and regional agencies in the Northeast may attempt to identify deficiencies in surveillance by state or subregion.

Connecticut, Delaware, Maine, Maryland, Massachuse↗

USGS2013

Reduced Myxobolus cerebralis actinospore production in a Colorado reservior may be linked to changes in Tubifex tubifex population structure

Elucidating the dynamics of a parasitic infection requiring two hosts in a natural ecosystem can be a daunting task. Myxobolus cerebralis (Mc), the myxozoan parasite that causes whirling disease in some salmonids, was detected in the Colorado River upstream of Windy Gap Reservoir (WGR) in 1988. Subsequently, whirling disease was implicated in the decline of wild Rainbow Trout Oncorhynchus mykiss in the river when WGR was identified as a point source of Mc triactinomyxons (TAMs). Between 1997 and 2004, numerous investigations began to elucidate the etiology of Mc in WGR. During this period, Mc TAM production in WGR declined more than 90%. Explanations for the decline have included differences in stream discharge between years, changes in the thermal regime of the lake, severe drought, changes in the fish population structure in WGR, and reductions in the prevalence and severity of Mc infection in salmonids in the Colorado and Fraser rivers upstream of WGR. All of these have been discredited as explanations for the reduced TAM production. In 2005, a new study was conducted to replicate the studies completed in 1998. In this paper, the results of a new real-time polymerase chain reaction assay utilized to quantify the mitochondrial 16S rDNA specific to each of four lineages of Tubifex tubifex in pooled samples of 50 oligochaetes are presented. These results suggest that compared with 1998, the densities of aquatic oligochaetes and T. tubifex have increased, TAM production has been greatly reduced, and the decline is congruent with the dominance of lineages I, V, and VI of T. tubifex—three lineages that are refractory or highly resistant to Mc infection—in the oligochaete population. While it is possible that the resistant lineages function as biofilters that deactivate Mc myxospores, the reason for the decline in TAM production in WGR remains an enigma.

Colorado↗