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USGS2013

Estimating transmission of avian influenza in wild birds from incomplete epizootic data: implications for surveillance and disease spreac

Estimating disease transmission in wildlife populations is critical to understand host–pathogen dynamics, predict disease risks and prioritize surveillance activities. However, obtaining reliable estimates for free-ranging populations is extremely challenging. In particular, disease surveillance programs may routinely miss the onset or end of epizootics and peak prevalence, limiting the ability to evaluate infectious processes. We used profile likelihood to estimate the force of infection (FOI) in a low pathogenic avian influenza virus (LPAIv) epizootic model from censored time series of LPAIv prevalence in hatch-year waterfowl (order Anseriformes) at postbreeding and migration sites in North America. We found a mean LPAIv FOI of 0·12 day −1 [95% CI, 0·00–0·39], corresponding to an incidence rate of 0·11 day −1 , with geographic heterogeneity (min–max: 0·02–0·23 day −1 ) among study sites. These high infection rates indicate that most hatch-year waterfowl are likely infected with LPAIv early in the fall migration. Comparison of model-predicted and observed immunity confirmed our assumption of naïve hatch-year waterfowl and suggested long-term immunity (>6 months) for adults. Using the mean LPAIv incidence rate, we predict a shorter and lower epizootic curve for highly pathogenic avian influenza virus (HPAIv; 5 weeks with peak prevalence of 28% and 30% mortality) than LPAIv (8 weeks with peak prevalence of 50%). These findings indicate it is harder to detect HPAIv than LPAIv with swabs from live birds, which are commonly used during disease surveillance. Synthesis and applications . Our study highlights the potential of integrating incomplete surveillance data with epizootic models to quantify disease transmission and immunity. This modelling approach provides an important tool to understand spatial and temporal epizootic dynamics and inform disease surveillance. Our findings suggest focusing highly pathogenic avian influenza virus (HPAIv) surveillance on postbreeding areas where mortality of immunologically naïve hatch-year birds is most likely to occur, and collecting serology to enhance HPAIv detection. Our modelling approach can integrate various types of disease data facilitating its use with data from other surveillance programs (as illustrated by the estimation of infection rate during an HPAIv outbreak in mute swans Cygnus olor in Europe).

Journal of Applied Ecology↗

USGS2012

Efficacy of a glycoprotein DNA vaccine against viral haemorrhagic septicaemia (VHS) in Pacific herring, Clupea pallasii Valenciennes

Viral haemorrhagic septicaemia virus (VHSV) and its associated disease state, viral haemorrhagic septicaemia (VHS), is hypothesized to be a proximate factor accounting for the decline and failed recovery of Pacific herring populations in Prince William Sound, AK (Marty et al. 1998, 2003, 2010). Survivors of laboratory-induced VHSV epizootics develop resistance to subsequent viral exposure (Kocan et al. 2001; Hershberger et al. 2007, 2010), which is likely the result of immune system recognition of the viral glycoprotein (G) (Lecocq-Xhonneux et al. 1994), a surface antigen that contains neutralizing epitopes (Lorenzen, Olesen & Jorgensen 1990; Jørgensen et al. 1995) and cell attachment domains (Lecocq-Xhonneux et al. 1994; Estepa & Coll 1996). These properties have proven useful in the development of G-gene-based DNA vaccines for VHSV and a related rhabdovirus, infectious haematopoietic necrosis virus (IHNV) (Anderson et al. 1996; Heppell et al. 1998; Corbeil et al. 1999; Einer-Jensen et al. 2009). Rainbow trout fingerlings, Oncorhynchus mykiss (Walbaum), vaccinated with 1 µg of either the VHS or IHN vaccine are protected from VHS when exposed to virus as early as 4 days (44 degree days) post-vaccination (p.v.) (Lorenzen et al. 2002). At later time points (80 days p.v.; 880 degree days), the level of cross-protection against VHS by IHN vaccination is either completely lost (60 days p.v.; 660 degree days) (3 g rainbow trout; 1 µg vaccine dose) (Lorenzen et al. 2002) or present at intermediate levels (6.5 g rainbow trout; 1 µg vaccine dose) (Einer-Jensen et al. 2009). Comparatively, VHS vaccination remains effective as long as 9 months (2520 degree days) p.v. (100 g rainbow trout; 0.5 µg vaccine dose) (McLauchlan et al. 2003). These results suggest that IHN and VHS vaccination activate a rapid transitory innate immune response against VHSV that is followed by long-term adaptive immunity in VHS-vaccinated trout (Lorenzen et al. 2002).

Journal of Fish Diseases↗

USGS1995

Detection and identification of aquatic birnaviruses by PCR assay

A reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for the detection and identification of aquatic birnaviruses. The four sets of primers (PrA, PrB, PrC, and PrD) that we used are specific for regions of cDNA coded by genome segment A of aquatic birnaviruses. PrA identifies a large fragment (1,180 bp) within the pVP2-coding region, and PrB identifies a 524-bp fragment within the sequence amplified by PrA. Primer set PrC frames a genome fragment (339 bp) within the NS-VP3-coding region, and PrD identifies a 174-bp sequence within the fragment identified by PrC. PrB and PrD amplified cDNAs from all nine recognized serotypes of aquatic birnavirus serogroup A as well as the N1 isolate that may represent a 10th serotype. These results indicate that these three primer sequences are highly conserved and can be used in PCR assays for group identification of these viruses. PrA routinely produced amplification products from eight serotypes but exhibited variable results with one serotype, and primer PrC identified 6 of the 11 virus isolates tested. The qualitative sensitivity of the RT-PCR assay was evaluated by comparison of the results with those of cell culture isolation assays. With the exception of one sample, the RT-PCR assay with primer PrD was as accurate as cell culture isolation for detecting virus in kidney and spleen tissues from naturally infected, asymptomatic carrier fish. These results indicate that the RT-PCR assay can be a rapid and reliable substitute for cell culture methods for the detection of aquatic birnaviruses.

Journal of Clinical Microbiology↗

USGS2020

Virome of bat guano from nine northern California roosts

Bats are hosts to a large variety of viruses, including many capable of cross species transmissions to other mammals or humans. We characterized the virome in guano from five common bat species in 9 Northern California roosts and a pool of 5 individual bats. Genomes belonging to 14 viral families known to infect mammals and 17 viral families infecting insects or of unknown tropism were detected. Near or complete genomes of a novel parvovirus, astrovirus, nodavirus, CRESS-DNA viruses and densoviruses and more partial genomes of a novel alphacoronavirus, and bunyavirus were characterized. Lower numbers of reads with >90% amino acid identity to previously described calicivirus, circovirus, adenoviruses, hepatovirus, bocaparvoviruses, and polyomavirus in other bat species were also found likely reflecting their wide distribution among different bats. Unexpectedly a few sequence reads of canine parvovirus 2 and the recently described mouse kidney parvovirus were also detected and their presence confirmed by PCR possibly originating from guano contamination by carnivores and rodents. The majority of eukaryotic viral reads were highly divergent indicating that numerous viruses still remain to be characterized even from such a heavily investigated order as Chiroptera.

California↗

USGS1978

Herpesvirus salmonis: Characterization of a new pathogen of rainbow trout

A new agent, provisionally designated Herpesvirus salmonis , was isolated from post-spawning rainbow trout ( Salmo gairdneri ) and studied primarily in the RTG-2 rainbow trout cell line. Infection of RTG-2 cells resulted in the formation of syncytia and Cowdry type A intranuclear inclusions. Replication occurred regularly at 5 and 10°C, but was inconsistent at 15°C, largely inhibited at 0°C, and completely inhibited at 20°C or higher. The virus was acid, heat, ether, and chloroform labile, but stable to freezing and thawing. It did not hemagglutinate. Viral DNA had a buoyant density of 1.709 g/cm 3 and a guanine-cytosine value of 50%. Hexagonal nucleocapsids had a diameter of 90 nm and were first seen in nuclei at 36 h. Enveloped forms measured about 150 nm and occurred both cytoplasmically and extracellularly. At 10°C, a one-step growth culture required about 96 h; cell-associated virus peaked at about 10 5 PFU/ml and exceeded released virus by a factor of about 10.

Journal of Virology↗

USGS2016

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↗

USGS2024

Identifying an understudied interface: Preliminary evaluation of the use of retention ponds on commercial poultry farms by wild waterfowl

While the recent incursion of highly pathogenic avian influenza into North America has resulted in notable losses to the commercial poultry industry, the mechanism by which virus enters commercial poultry houses is still not understood. One theorized mechanism is that waterfowl shed virus into the environment surrounding poultry farms, such as into retention ponds, and is then transmitted into poultry houses via bridge species. Little is known about if and when wild waterfowl use these retention ponds, leading to uncertainty regarding the potential significance of this interface. To quantify the use of retention ponds on commercial poultry farms by wild waterfowl, we surveyed 12 such ponds across Somerset and Dorchester counties, Maryland, USA. This region was chosen due to the high level of poultry production and its importance for migratory waterfowl. Surveys consisted of recording waterfowl visible on the retention ponds from public roadways at least once per week from 20 September 2022–31 March 2023. Throughout the course of this study, we observed a total of nine species of waterfowl using retention ponds on commercial poultry farms at nine of 12 sites. The number of waterfowl observed at retention ponds varied notably throughout the course of our survey period, with values generally following trends of fall migration within each species indicating that resident birds were not the only individuals to utilize these habitats. Additionally, waterfowl use was highest at sites with little vegetation immediately surrounding the pond, and lowest when ponds were surrounded by trees. Our data suggest that retention ponds on commercial poultry farms present a notable interface for waterfowl to introduce avian influenza viruses to farm sites. However, additional testing and surveys could provide further insight into whether it may be possible to reduce the use of these habitats by wild waterfowl through vegetative management as preliminarily reported here.

Transboundary and Emerging Diseases↗

USGS2022

A novel herpesvirus detected in 3 different species of chelonians

Herpesviruses are found in free-living and captive chelonian populations, often in association with morbidity and mortality. To date, all known chelonian herpesviruses fall within the subfamily Alphaherpesvirinae . We detected a novel herpesvirus in 3 species of chelonians: a captive leopard tortoise ( Stigmochelys pardalis ) in western TX, USA; a steppe tortoise ( Testudo [ Agrionemys ] horsfieldii ) found near Fort Irwin, CA, USA; and 2 free-living, three-toed box turtles ( Terrapene mexicana triunguis ) found in Forest Park, St. Louis, MO. The leopard tortoise was coinfected with the tortoise intranuclear coccidian and had clinical signs of upper respiratory tract disease. The steppe tortoise had mucopurulent nasal discharge and lethargy. One of the three-toed box turtles had no clinical signs; the other was found dead with signs of trauma after being observed with blepharedema, tympanic membrane swelling, cervical edema, and other clinical signs several weeks prior to death. Generally, the branching order of the turtle herpesviruses mirrors the divergence patterns of their hosts, consistent with codivergence. Based on phylogenetic analysis, this novel herpesvirus clusters with a clade of viruses that infect emydid hosts and is likely of box turtle origin. Therefore, we suggest the name terrapene alphaherpesvirus 3 (TerAHV3) for the novel virus. This virus also has the ability to host-jump to tortoises, and previously documented herpesviral morbidity tends to be more common in aberrant hosts. The relationship between clinical signs and infection with TerAHV3 in these animals is unclear, and further investigation is merited.

Journal of Veterinary Diagnostic Investigation↗

USGS2007

Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

Background The role of wild birds in the dispersal of highly pathogenic avian influenza virus H5N1 continues to be the subject of considerable debate. However, some researchers functionally examining the same question are applying opposing null hypotheses when examining this issue. Discussion I describe the correct method for establishing a null hypothesis under the scientific method. I suggest that the correct null hypothesis is that migratory birds can disperse this virus during migration and encourage researchers to design studies to falsify this null. Finally, I provide several examples where statements made during this debate, while strictly true, are not generally informative or are speculative. Summary By adhering to the scientific method, definitive answers regarding the role of wild birds in the dispersal of highly pathogenic viruses will be reached more effectively.

Virology Journal↗

USGS2019

Influenza A prevalence and subtype diversity in migrating teal sampled along the United States Gulf Coast

Wild birds in the order Anseriformes are important reservoirs for influenza A viruses (IAV); however, IAV prevalence and subtype diversity may vary by season, even at the same location. To better understand the ecology of IAV during waterfowl migration through the Gulf Coast of the United States (Louisiana and Texas), surveillance of blue-winged (Spatula discors) and American green-winged (Anas carolinensis) teal was conducted annually during the spring (live-capture; 2012-2017) and fall (hunter-harvested; 2007-2017) at times inferred to coincide with northward and southward movements, respectively, for these waterfowl species. During spring migration, 266 low pathogenicity (LP) IAV positive samples were recovered from 7,547 paired cloacal/oropharyngeal (COP) samples (prevalence: 3.5%; annual range: 1.3%-8.4%). During fall migration, 650 LP IAV positive samples were recovered from 9,493 COP samples (prevalence: 6.8%; annual range: 0.4%-23.5%). Overall, 34 and 20 different IAV subtypes were recovered during fall and spring sampling, respectively. Consistent with previous results for fall migrating ducks, H3 and H4 HA subtypes were most common; however, H4 subtype viruses predominated every year. This is in contrast to the predominance of LP H7 and H10 HA subtype viruses in both species during spring. The N6 and N8 NA subtypes, which were usually associated with H4, were most common during fall; the N6 subtype was not recovered in the spring. These consistent seasonal trends in IAV subtype detection in both species are currently not understood and highlight the need for further research regarding potential drivers of spatiotemporal patterns of infection such as population immunity.

Avian Diseases↗

USGS2010

Rice production systems and avian influenza: Interactions between mixed-farming systems, poultry and wild birds

Wild waterfowl are the reservoir for avian influenza viruses (AIVs), a family of RNA viruses that may cause mild sickness in waterbirds. Emergence of H5N1, a highly pathogenic avian influenza (HPAI) strain, causing severe disease and mortality in wild birds, poultry and humans, had raised concerns about the role of wild birds in possible transmission of the disease. In this review, the link between rice production systems, poultry production systems, and wild bird ecology is examined to assess the extent to which these interactions could contribute towards the persistence and evolution of HPAI H5N1. The rice (Oryza sativa) and poultry production systems in Asia described, and then migration and movements of wild birds discussed. Mixed farming systems in Asia and wild bird movement and migration patterns create opportunities for the persistence of low pathogenic AIVs in these systems. Nonetheless, there is no evidence of long-term persistence of HPAI viruses (including the H5N1 subtype) in the wild. There are still significant gaps in the understanding of how AIVs circulate in rice systems. A better understanding of persistence of AIVs in rice farms, particularly of poultry origins, is essential in limiting exchange of AIVs between mixed-farming systems, poultry and wild birds.

Waterbirds↗

USGS2025

Serological assessment of pathogen exposure among desert bighorn sheep in southwestern Arizona

Bighorn sheep ( Ovis canadensis ) are influenced by infectious diseases. Although Mycoplasma ovipneumoniae has been the main focus of bighorn sheep managers since early 2010, other pathogens may also influence bighorn sheep populations. We sampled desert bighorn sheep ( Ovis canadensis mexicana ) captured for a study on the Cabeza Prieta National Wildlife Refuge in southwestern Arizona, USA, 2001–2005, for a suite of pathogens: bluetongue (BT), epizootic hemorrhagic disease (EHD), parainfluenza 3, bovine respiratory syncytial virus, Clamydia , seven Leptospira serovars, bovine herpesvirus 1 (causative agent of infectious bovine rhinotracheitis), bovine viral diarrhea virus, and bovine respiratory syncytial virus. We recorded evidence of exposure to seven of these pathogens, with two Leptospira serovars ( hardjo and bratislava ), five strains of BT, and two strains of EHD detected. Seroprevalence rates of detected pathogens varied between 12–49%. We observed high levels of co-occurrence for EHD and BT. These results highlight that multiple pathogens may influence desert bighorn sheep populations. These data also provide historical context to pathogen exposure for a region where few such data are available.

Arizona↗

USGS1983

Inclusion body disease of cranes: A serological follow-up to the 1978 die-off

A herpesvirus was isolated from captive cranes involved in a 1978 die-off. Neutralizing antibody to this virus was detected in this captive population as early as 1975 and consistently thereafter through 1979. Exposure to the virus evidently occurred at least 2 1/2 years before the die-off, without causing any mortality diagnosed as being caused by inclusion body disease of cranes (IBDC). Overcrowding and environmental conditions in 1978 may have contributed to the deaths of certain species of cranes in one area and not in another. Mortality ratios and serological data suggest that crane species vary in their response to IBDC virus.

Wisconsin↗

USGS2016

U.S. Geological Survey science strategy for highly pathogenic avian influenza in wildlife and the environment (2016–2020)

Introduction Through the Science Strategy for Highly Pathogenic Avian Influenza (HPAI) in Wildlife and the Environment, the USGS will assess avian influenza (AI) dynamics in an ecological context to inform decisions made by resource managers and policymakers from the local to national level. Through collection of unbiased scientific information on the ecology of AI viruses and wildlife hosts in a changing world, the U.S. Geological Survey (USGS) will enhance the development of AI forecasting tools and ensure this information is integrated with a quality decision process for managing HPAI. The overall goal of this USGS Science Strategy for HPAI in Wildlife and the Environment goes beyond documenting the occurrence and distribution of AI viruses in wild birds. The USGS aims to understand the epidemiological processes and environmental factors that influence HPAI distribution and describe the mechanisms of transmission between wild birds and poultry. USGS scientists developed a conceptual model describing the process linking HPAI dispersal in wild waterfowl to the outbreaks in poultry. This strategy focuses on five long-term science goals, which include: Science Goal 1—Augment the National HPAI Surveillance Plan; Science Goal 2—Determine mechanisms of HPAI disease spread in wildlife and the environment; Science Goal 3—Characterize HPAI viruses circulating in wildlife; Science Goal 4—Understand implications of avian ecology on HPAI spread; and Science Goal 5—Develop HPAI forecasting and decision-making tools. These goals will help define and describe the processes outlined in the conceptual model with the ultimate goal of facilitating biosecurity and minimizing transfer of diseases across the wildlife-poultry interface. The first four science goals are focused on scientific discovery and the fifth goal is application-based. Decision analyses in the fifth goal will guide prioritization of proposed actions in the first four goals.

Open-File Report↗

USGS2004

Fecal-indicator bacteria in the Newfound Creek watershed, western North Carolina, during a high and low streamflow condition, 2003

Water quality in the Newfound Creek watershed has been shown to be affected by bacteria, sediment, and nutrients. In this study, Escherichia coli (E. coli) bacteria were sampled at five sites in Newfound Creek and five tributary sites during low flow on May 28, 2003, and high flow on November 19, 2003. In addition, a subset of five sites was sampled for fecal coliform bacteria, E. coli bacteria in streambed sediments (low flow only), and coliphage virus for serotyping. Coliphage virus serotyping has been used to identify human and animal sources of bacterial contamination. A streamflow gage was installed and operated to support ongoing water-quality studies in the watershed. Fecal coliform densities ranged from 92 to 27,000 colony-forming units per 100 milliliters of water for E. coli and 140 to an estimated 29,000 colony-forming units per 100 milliliters of water for fecal coliform during the two sampling visits. Ninety percent of the E. coli and fecal coliform samples exceeded corresponding U.S. Environmental Protection Agency or North Carolina water-quality criteria for recreational and ambient waters. During low flow, the middle part of the Newfound Creek watershed and the Dix Creek tributary had the highest densities of E. coli bacteria. During the high-flow sampling, all tributaries contained high densities of E. coli bacteria, although Dix Creek and Round Hill Branch were the largest contributors of these bacteria to Newfound Creek. Coliphage virus serotyping results were inconclusive because most samples did not contain the male-specific RNA coliphage needed for serotyping. Positive results indicated, however, that during low flow, non-human sources of bacteria were present in Sluder Branch, and during high flow, human sources of bacteria were present in Round Hill Branch. Sampling of bacteria in streambed sediments during low flow indicated that sediments do not appear to be a substantial source of bacteria relative to the water column, with the exception of an area near the confluence of Sluder Branch and Newfound Creek.

North Carolina↗

USGS2000

Microbiological and chemical quality of ground water used as a source of public supply in southern Missouri — Phase II, April–July, 1998

The protection of public health through quality public ground-water systems is the responsibility of the U. S. Environmental Protection Agency and the State of Missouri, through the Missouri Department of Natural Resources, Public Drinking Water Program. Approximately 95 percent of the public-water supplies in Missouri use ground water as their source of drinking water through more than 3,700 public wells. Karst terrain, intensive agricultural operations, extensive numbers of on-site sewage systems, and poor well construction can lead to chemical and microbiological contamination of the contributing aquifers. Sitespecific studies and routine regulatory monitoring have produced information on the overall quality and potability of the State's public-drinking-water supplies, but little is known about the presence of viruses. The U.S. Geological Survey, in cooperation with the Missouri Department of Natural Resources, sampled 109 public-water supplies to characterize the physical, chemical, bacterial, and viral conditions in southern Missouri. During April to July 1998, these wells were sampled for nutrients, total organic carbon, optical brighteners, indicator bacteria, enteric viruses, and ribonucleic acid and somatic coli phages. These constituents indicate possible surface contamination of the sampled aquifer. Selection of the wells to be sampled depended on the age of the well (pre-1970), land use, geohydrology, and well construction. None of the physical or chemical constituents measured or analyzed exceeded Missouri's Drinking Water Standards set by the Public Drinking Water Program of the Missouri Department of Natural Resources. The majority of ammonia plus organic nitrogen, nitrite, and phosphorus concentrations were below the laboratory's minimum reporting levels. There were a greater number of detects above the minimum reporting level with respect to the nitrite plus nitrate, ammonia, orthophosphate, and total organic carbon concentrations. Analyses included comparing and contrasting the data by grouping according to well age and construction, karst type, geohydrology, soil type, and land use. There was little variation in well construction between selected wells. The results indicated several groupings of similar and dissimilar concentrations, most expected because of hydrological, physical, or land use differences. Dissolved oxygen values indicated distinct variation in the different groupings. There were significant differences in dissolved oxygen values between the secondary and non-karst areas, the Ozark confined and Ozark unconfined geohydrologic groups, and between agricultural and other land uses. In groupings by soil and geohydrology, the Missouri bootheel region differed with respect to ammonia, total organic carbon, and phosphorus when compared with the other groups. Less than 10 percent of the wells sampled tested positive for bacterial contamination. E. coli was the most frequently detected bacterium. The public wells at Monett and West Plains, Missouri, had plates with colonies too numerous to count for all three indicator bacteria. Further analyses by rRNA (ribosomal RiboNucleic Acid) hybridization techniques detennined that much of the bacteria present were from ruminant and human sources. No enteric viruses were detected in the 109 samples. Both ribonucleic acid and somatic coliphage were detected at two wells. One additional well had ribonucleic acid coliphage and another had somatic coliphage for a total of four wells with coliphage selects.

Missouri↗

USGS1993

Determination of buoyant density and sensitivity to chloroform and freon for the etiological agent of infectious salmonid anaemia

Plasma was collected from Atlantic salmon Salrno salar with acute infectious salmon anaemia (ISA) and used to challenge Atlantic salmon parr by intraperitoneal injection. Treatment of plasma with the lipid solvent, chloroform, showed that the etiological agent of ISA contained essential lipids, probably as a viral envelope. Some infectivity remained following treatment with freon. Injection challenges using fractions from equilibrium density gradient centrifugation of plasma from fish with acute ISA revealed a band of infectivity in the range 1.184 to 1.262 g cm-3. The band was believed to conta~n both complete ISA-virus particles and infectious particles lacking a complete envelope, nucleocapsid or genome. Density gradient centrifugation of infectious plasma for enrichment of the putative ISA virus appeared to offer a suitable method for obtaining virus-specific nucleic acid for use in the construction of cDNA libraries.

Diseases of Aquatic Organisms↗

USGS2010

Establishment and partial characterization of a cell line from burbot Lota lota maculosa: susceptibility to IHNV, IPNV and VHSV.

This study describes the development and partial characterization of a continuous fibroblastic-like cell line (BEF-1) developed from late stage embryos of North American burbot Lota lota maculosa. This cell line has been maintained for over 5 yr and 100 passages in vitro. Cells were cultured using Eagle’s minimum essential medium with Earle’s salts (MEM) supplemented with GlutaMAX™, and 10% fetal bovine serum (FBS), pH 7.4. The addition of penicillin-streptomycin-neomycin (PSN) antibiotic mixture (0.05, 0.05, 0.1 mg ml–1, respectively) did not negatively influence cell replication; however, the antimycotic Fungizone™ (2.5 µg ml–1, amphotericin B) caused cell rounding and resulted in a severe decrease in cell proliferation. Optimal incubation temperature has been observed between 15 and 23°C, and at these temperatures cultures are routinely passed using standard trypsinization methods every 5 to 7 d at a split ratio of 1:3 or 1:4. The cell line was susceptible to isolates of the M and U North American genotypes of infectious hematopoietic necrosis virus (IHNV), and to isolates of genotypes I, IVa, and IVb of viral hemorrhagic septicemia virus (VHSV). In contrast, the cell line was refractory to infection by 2 North American isolates of infectious pancreatic necrosis virus (IPNV) from serotypes A1 and A9. This cell line provides a new laboratory tool, will allow further investigation into viral diseases of burbot and possibly other species, and is the first immortalized cell line reported from a species in the Gadidae (cod) family.

Diseases of Aquatic Organisms↗