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The representative research animal: Why rainbow trout (Salmo gairdneri Rich.)?

In laboratory research, the rainbow trout has become a counterpart to the white rat, because that fish is an adaptable species available in much of the developed world and stocks from egg through adult are available throughout the year. Moreover, many strains are recognized, and their propagation and laboratory maintenance are not particularly demanding. Also, knowledge of rainbow trout nutrition, husbandry, diseases, immune responses, toxicology, and carcinogenesis exceeds that of any other salmonid or coldwater teleost. The rainbow trout is the logical surrogate species in many studies of other salmonids.

Journal of Applied Ichthyology

Detection of Viral Hemorrhagic Septicemia Virus by Quantitative Reverse Transcription Polymerase Chain Reaction from Two Fish Species at Two Sites in Lake Superior

Viral hemorrhagic septicemia virus (VHSV) was first detected in the Laurentian Great Lakes in 2005 during a mortality event in the Bay of Quinte, Lake Ontario. Subsequent analysis of archived samples determined that the first known isolation of VHSV in the Laurentian Great Lakes was from a muskellunge Esox masquinongy collected in Lake St. Clair in 2003. By the end of 2008, mortality events and viral isolations had occurred in all of the Laurentian Great Lakes except Lake Superior. In 2009, a focused disease surveillance program was designed to determine whether VHSV was also present in Lake Superior. In this survey, 874 fish from 7 sites along the U.S. shoreline of Lake Superior were collected during June 2009. Collections were focused on nearshore species known to be susceptible to VHSV. All fish were dissected individually by using aseptic techniques and were tested for the presence of VHSV genetic material by use of a quantitative reverse transcription (qRT) polymerase chain reaction (PCR) targeting the viral nucleoprotein gene. Seventeen fish from two host species at two different sites tested positive at low levels for VHSV. All attempts to isolate virus in cell culture were unsuccessful. However, the presence of viral RNA was confirmed independently in five fish by using a nested PCR that targeted the glycoprotein (G) gene. Partial G gene sequences obtained from three fish were identical to the corresponding sequence from the original 2003 VHSV isolate (MI03) from muskellunge. These detections represent the earliest evidence for the presence of VHSV in Lake Superior and illustrate the utility of the highly sensitive qRT-PCR assay for disease surveillance in aquatic animals.

Journal of Aquatic Animal Health

Susceptibility of Koi and Yellow Perch to infectious hematopoietic necrosis virus by experimental exposure

Infectious hematopoietic necrosis virus (IHNV) is a novirhabdoviral pathogen that originated in western North America among anadromous Pacific salmonids. Severe disease epidemics in the late 1970s resulting from IHNV's invasion into farmed Rainbow Trout Oncorhynchus mykiss in North America, Asia, and Europe emphasized IHNV's ability to adapt to new hosts under varying rearing conditions. Yellow Perch Perca flavescens and Koi Carp Cyprinus carpio (hereafter, “Koi”) are aquaculture-reared fish that are highly valued in sport fisheries and the ornamental fish trade, respectively, but it is unknown whether these fish species are vulnerable to IHNV infection. In this study, we exposed Yellow Perch, Koi, and steelhead (anadromous Rainbow Trout) to IHNV by intraperitoneal injection (10 6 PFU/fish) and by immersion (5.7×10 5 PFU/mL) for 7 h, and monitored fish for 28 d. The extended immersion exposure and high virus concentrations used in the challenges were to determine if the tested fish had any level of susceptibility. After experimental exposure, Yellow Perch and Koi experienced low mortality (<6%) compared with steelhead (>35%). Virus was found in dead fish of all species tested and in surviving Yellow Perch by plaque assay and quantitative reverse transcription polymerase chain reaction (qPCR), with a higher prevalence in Yellow Perch than Koi. Infectious virus was also detected in Yellow Perch out to 5 d after bath challenge. These findings indicate that Yellow Perch and Koi are highly resistant to IHNV disease under the conditions tested, but Yellow Perch are susceptible to infection and may serve as possible virus carriers.

Journal of Aquatic Animal Health

Development of a multiplex fluorescence in situ hybridization assay to identify coinfections in young-of-the-year smallmouth bass

Histopathological assessments of young-of-the-year (age-0) Smallmouth Bass Micropterus dolomieu in the Susquehanna River drainage identified a high prevalence of the myxozoan Myxobolus inornatus . This myxozoan infects the connective tissue of the muscle below the skin but is sometimes observed in the esophagus and buccal cavity. In some instances, shallow infections cause breaks in the skin, which could increase the chance of opportunistic bacterial infections. Several microbial pathogens, including Flavobacterium columnare , Aeromonas spp., and Largemouth Bass virus, have also been cultured from clinically diseased young of year. A multiplex fluorescence in situ hybridization (FISH) assay was developed to determine potential colocalization of M. inornatus , Flavobacterium spp., and Aeromonas spp. infections. With FISH, 75% of age-0 Smallmouth Bass exhibited M. inornatus infections, 10% had Aeromonas spp. infections, and 5% had Flavobacterium spp. infections, while 3% had coinfections with both bacterial species and M. inornatus . The results of the multiplex FISH assay revealed a low occurrence of coinfections of Flavobacterium spp. and/or Aeromonas spp. with M. inornatus in randomly sampled individuals.

Pennsylvania

An acute septicaemic disease of brown trout (Salmo trutta) and Atlantic salmon (Salmo salar) caused by a Pasteurella-like organism

Ulcerations of the skin associated with haemorrhagic petechiae of liver and kidneys, were the main signs of disease affecting salmon and brown trout in Norway. A death rate of 15–20% was estimated for the 5 month period of mid-March-August, although mortalities occurred throughout the year. Bacteriological examinations, involving 36 isolates, suggested the causual organism to be a Pasteurella although the DNA homology examination gave a G.C. ratio of 55.6 % which is high for Pasteurella as a group.

Journal of Fish Biology

Characteristics and identification of oxidative Pseudomonads isolated from diseased fish

Several characteristics of 20 strains of oxidative pseudomonads, all but one of which were isolated from freshwater fish, were determined. All strains oxidized glucose and gluconate, produced NH 3 from 1 % peptone, grew at 6, 12, 20 and 30° and produced cytochrome oxidase. Lipase was produced by 17 of the strains, while 15 produced gelatinase, and 16 produced fluorescent pigment. These and other characteristics used to identify fish-spoilage pseudomonads are also useful for identification of fish-pathogenic pseudomonads. Species determinations from the present data were not attempted. However, from previously published data on speciation of Pseudomonas , most of the 20 strains appeared to be closely related to or identical with P. fluorescens .

Journal of General Microbiology

Gross and microscopic pathology of lesions in Pocillopora spp. from the subtropical eastern Pacific

Coral reefs are threatened by a variety of factors including diseases that have caused significant damage in some regions such as in the Caribbean. At present, no data are available on coral diseases in the Mexican Pacific where Pocillopora spp. is a dominant component of coral communities. Here, we describe gross and microscopic morphology of lesions found in pocilloporids at four sites in the Mexican Pacific. Corals were identified and their lesions photographed and quantified in the field. Tissue samples were collected from healthy and affected colonies for histopathology. We recorded seven species of pocilloporids at the study sites with Isla Isabel being the location with the highest coral diversity ( H ′ = 1.27). Lesions were present in 42% of the colonies and included discoloration (32%), predation-induced tissue loss (30%), unexplained tissue loss (3%) and overgrowth by sponges or algae (35%). The most affected species, P . damicornis (50%), was also one of the most common in the region. No species was more prone to a particular lesion, but there was a significant association between location and the presence of lesions. Northern Islas Marietas (61%) and Isla Isabel (41%) had the highest prevalence of lesions, followed by Manzanillo (37%) and Bahías de Huatulco (23%). Histological changes included atrophy of the surface body wall with depletion of zooxanthellae (91%) in corals with discoloration (bleaching). Ablation of tissue from mesoglea (18%) was also observed. Colonies with unexplained tissue loss showed atrophy and thinning of the epidermis (89%), characterized by cuboidal instead of pseudocolumnar cells normally found in healthy pseudocolumnar ciliated epithelium. Bacterial aggregates between the mesoglea and gastrodermis (11%) were very conspicuous in healthy and diseased corals. Lesions produced by fish bites and gastropods were associated with tissue atrophy (40%) and, in some cases, algal overgrowth near the lesion (20%). No infectious agents associated with cell pathology were detected microscopically. Bleaching and overgrowth by algae and sponges, as well as unexplained tissue loss, are common in Pocillopora . These lesions and anatomical changes warrant further study since their incidence is potentially indicative of reef degradation.

Mexican Pacific

Immunostimulants in fish diets

Various immunostimulants and their methods of application in fish culture are examined in this review. Important variables such as life stage and innate disease resistance of the fish; immunostimulant used, its structure and mode of action; and the fish's environment are discussed. Conflicting results have been published about the efficacy of immunostimulants in fish diets. Some researchers have had positive responses demonstrated as increased fish survival, others have not. Generally, immunostimulants enhance individual components of the non-specific immune response but that does not always translate into increased fish survival. In addition, immunostimulants fed at too high a dose or for too long can be immunosuppressive. [Article copies available for a fee from The Haworth Document Delivery Service: 1-800-342-9678. E-mail address: getinfo@haworthpressinc.com ].

Journal of Applied Aquaculture

Fish viruses: Isolation and identification of infectious hematopoietic necrosis in eastern North America

Infectious hematopoietic necrosis virus (IHNV) was isolated from diseased fingerling rainbow trout ( Salmo gairdneri ) from a hatchery in West Virginia. Clinical signs, histopathologic findings, and origin of eggs provided a basis for diagnosis, and virus was isolated and presumptively identified by plaque characteristics. Serum neutralization tests provided positive identification of the agent as IHNV, and electronmicroscopy showed its rhabdovirus morphology. Experimental infections resulted in signs of IHNV and death; test fish had characteristic histopathologic alterations and appropriate virus titers. This is the first completely documented occurrence of IHNV beyond the Pacific Northwest.

Journal of the Fisheries Research Board of Canada

Quantitative and qualitative studies of gut flora in striped bass from estuarine and coastal marine environments

Examination of the intestinal contents of 130 striped bass ( Morone saxatilis ) collected from the Hudson River and Long Island Sound during May to October 1981 showed that opportunistic fish pathogens—especially Aeromonas hydrophila —predominated in samples from both locations. Other isolates from both groups of striped bass included Vibrio , pseudomonads, flavobacteria, Alcaligenes , and enterics. Small numbers of Micrococcus , Bacillus , Corynebacterium , and Acinetobacter were also isolated. Total numbers of bacteria in the intestines were 100 to 1,000 times higher in striped bass from the Hudson River than in those from Long Island Sound.

Journal of Wildlife Diseases

Mycobacteriosis in a black-tailed deer (Odocoileus hemionus columbianus) caused by Mycobacterium kansasii

An eviscerated hunter-harvested female black-tailed deer ( Odocoileus hemionus columbianus ) was submitted to the Washington Department of Fish and Wildlife. The deer was emaciated, devoid of adipose tissue, and the parietal surface of the thoracic cavity contained multiple granulomas. Acid-fast bacteria were detected histologically from the granulomas and were isolated and identified as Mycobacterium kansasii , a nontuberculous mycobacterium sporadically reported to cause tuberculosis-like disease in a variety of vertebrates. This was the first report of symptomatic disease caused by M. kansasii in free-ranging deer. This case indicates that atypical mycobacteria can cause tuberculosis-like disease in free-ranging deer and illustrates the importance of identifying causative agents of tuberculosis-like disease in wildlife.

Journal of Zoo and Wildlife Medicine

A novel approach for directly incorporating disease into fish stock assessment: A case study with seroprevalence data

When estimating mortality from disease with fish population models, common disease surveillance data such as infection prevalence are not always informative, especially for fast-acting diseases that may go unobserved in infrequently sampled populations. In these cases, seroprevalence — the proportion of fish with measurable antibody levels in their blood — may be more informative. In cases of life-long immunity, seroprevalence data require less frequent sampling intervals than infection prevalence data and can reflect the cumulative exposure history of fish. We simulation tested the usefulness of seroprevalence data in an age-structured fish stock assessment model using viral hemorrhagic septicemia virus (VHSV) in Pacific herring ( Clupea pallasii ) as a case study. We developed a novel epidemiological model to simulate population dynamics and seroprevalence data and fitted to these data in an integrated catch-at-age model with equations that estimate age- and time-varying mortality from disease. We found that simulated seroprevalence data can provide accurate estimates of infection history and disease-associated mortality. Importantly, even models that misspecified nonstationary processes in background or disease-associated mortality, but included seroprevalence data, accurately estimated annual infection and population abundance.

Canadian Journal of Fisheries and Aquatic Sciences

Control of whirling disease (Myxosoma cerebralis): Effects of drying, and disinfection with hydrated lime or chlorine

Thorough air drying of contaminated mud killed the spores of Myxosoma cerebralis . Treatment of simulated earthen ponds with hydrated lime with up to 4550 g/m 2 on the wet mud did not kill all of the spores. Treatment of simulated earthen ponds with up to 1200 p.p.m. chlorine did not kill all of the spores, however, treatment of contaminated water with 10 p.p.m. chlorine did kill the spores.

Journal of Fish Biology

Recovery of a fish pathogenic bacterium, Aeromonas salmonicida, from ebonyshell mussels Fusconaia ebena using nondestructive sample collection procedures

Refugia are increasingly being used to maintain and propagate imperiled freshwater mussels for future population augmentations. Success for this endeavor is dependent on good husbandry, including a holistic program of resource health management. A significant aspect to optimal health is the prevention or control of infectious diseases. Describing and monitoring pathogens and diseases in mussels involves examination of tissues or samples collected from an appropriate number of individuals that satisfies a certain confidence level for expected prevalences of infections. In the present study, ebonyshell mussels Fusconaia ebena were infected with a fish pathogenic bacterium, Aeromonas salmonicida, through their cohabitation with diseased brook trout Salvelinus fontinalis. At a 100% prevalence of infection, the F. ebena were removed from the cohabitation tank to clean tanks that were supplied with pathogen-free water, which initiated their depuration of A. salmonicida. Three samples (nondestructive fluid, mantle, hemolymph) collected using nondestructive procedures were compared with fluids and soft tissue homogenates collected after sacrificing the mussels for recovery of the bacterium during this period of depuration. Nondestructive sample collections, especially ND fluid, provide a comparable alternative to sacrificing mussels to determine pathogen status.

Journal of Shellfish Research

Renibacterium salmoninarum in spring-summer Chinook salmon smolts at dams on the Columbia and Snake Rivers

We evaluated Renibaeterium salmoninarum infection in smolts of hatchery and wild spring-summer Chinook salmon Oncorhynchus tshawytscha sampled during most of the outmigration at Little Goose (1988) and Lower Granite dams (1988–1991) on the Snake River and at Priest Rapids and McNary dams on the Columbia River (1988–1990). We sampled 860–2,178 fish per dam each year. Homogenates of kidney–spleen tissue from all fish were tested for the presence of R. salmoninarum antigens by the enzyme-linked immunosorbent assay (ELISA), and homogenates from 10% of the fish were examined by the fluorescent antibody technique (FAT). Although only 1–11% of fish sampled at a given dam during any l year exhibited lesions characteristic of bacterial kidney disease, 86–100% of the fish tested positive for R. salmoninarum antigen by ELISA, whereas 4–17% of the fish tested positive by the FAT. During most years, a majority (68–87%) of fish testing positive by the ELISA had low R. salmoninarum antigen levels, but in 1989, 53% of positive fish from Lower Granite Dam and 52% from McNary Dam showed medium-to-high antigen levels. For most years, the highest mean antigen levels were measured in fish sampled after 75% of the total out-migrants had passed a given dam. When the largest numbers of fish were being collected for bypass or downriver transportation, mean antigen levels were relatively low.

Washington

Fish viruses: A double-stranded RNA icosahedral virus from a North American cyprinid

A previously unreported virus disease of cultured golden shiners ( Notemigonus crysoleucas ) is described. The condition is called golden shiner virus (GSV) disease. The virus is icosahedral, measures approximately 70 nm, is ether and heat resistant, stable at pH 3, 7, and 10, and appears to have a double stranded RNA core. It was not neutralized by infectious pancreatic necrosis virus specific antiserum. GSV causes syncytial like cytopathic effect in fathead minnow cells but not brown bullhead, rainbow trout gonad (RTG-2) or channel catfish ovary cells. Severe mortalities in golden shiner culture ponds do not appear to be associated with GSV infections.

Journal of the Fisheries Research Board of Canada

Principles underlying the epizootiology of viral hemorrhagic septicemia in Pacific herring and other fishes throughout the North Pacific Ocean

Although viral hemorrhagic septicemia virus (VHSV) typically occurs at low prevalence and intensity in natural populations of Pacific herring ( Clupea pallasii ) and other marine fishes in the Northeast Pacific Ocean, epizootics of the resulting disease (VHS) periodically occur, often in association with observed fish kills. Here we identify a list of principles, based on a combination of field studies, controlled laboratory experiments, and previously unpublished observations, that govern the epizootiology of VHS in Pacific herring. A thorough understanding of these principles provides the basis for identifying risk factors that predispose certain marine fish populations to VHS epizootics, including the lack of population resistance, presence of chronic viral carriers in a population, copious viral shedding by infected individuals, cool water temperatures, limited water circulation patterns, and gregarious host behavioral patterns. Further, these principles are used to define the epizootiological stages of the disease in Pacific herring, including the susceptible (where susceptible individuals predominate a school or subpopulation), enzootic (where infection prevalence and intensity are often below the limits of reasonable laboratory detection), disease amplification (where infection prevalence and intensity increase rapidly), outbreak (often accompanied by host mortalities with high virus loads and active shedding), recovery (in which the mortality rate and virus load decline owing to an active host immune response), and refractory stages (characterized by little or no susceptibility and where viral clearance occurs in most VHS survivors). In addition to providing a foundation for quantitatively assessing the potential risks of future VHS epizootics in Pacific herring, these principles provide insights into the epizootiology of VHS in other fish communities where susceptible species exist.

Canadian Journal of Fisheries and Aquatic Sciences

Sex-biased gene flow among elk in the greater Yellowstone ecosystem

We quantified patterns of population genetic structure to help understand gene flow among elk populations across the Greater Yellowstone Ecosystem. We sequenced 596 base pairs of the mitochondrial control region of 380 elk from eight populations. Analysis revealed high mitochondrial DNA variation within populations, averaging 13.0 haplotypes with high mean gene diversity (0.85). The genetic differentiation among populations for mitochondrial DNA was relatively high ( F ST = 0.161; P = 0.001) compared to genetic differentiation for nuclear microsatellite data ( F ST = 0.002; P = 0.332), which suggested relatively low female gene flow among populations. The estimated ratio of male to female gene flow ( m m / m f = 46) was among the highest we have seen reported for large mammals. Genetic distance (for mitochondrial DNA pairwise F ST ) was not significantly correlated with geographic (Euclidean) distance between populations (Mantel's r = 0.274, P = 0.168). Large mitochondrial DNA genetic distances (e.g., F ST > 0.2) between some of the geographically closest populations (<65 km) suggested behavioral factors and/or landscape features might shape female gene flow patterns. Given the strong sex-biased gene flow, future research and conservation efforts should consider the sexes separately when modeling corridors of gene flow or predicting spread of maternally transmitted diseases. The growing availability of genetic data to compare male vs. female gene flow provides many exciting opportunities to explore the magnitude, causes, and implications of sex-biased gene flow likely to occur in many species.

Yellowstone National Park