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At least 631 records · Page 35Linked to original sources
USGS2004

Cloning, expression, cellular distribution, and role in chemotaxis of a C5a receptor in rainbow trout: The first identification of a C5a receptor in a nonmammalian species

C3a, C4a, and C5a anaphylatoxins generated during complement activation play a key role in inflammation. C5a is the most potent of the three anaphylatoxins in eliciting biological responses. The effects of C5a are mediated by its binding to C5a receptor (C5aR, CD88). To date, C5aR has only been identified and cloned in mammalian species, and its evolutionary history remains ill-defined. To gain insights into the evolution, conserved structural domains, and functions of C5aR, we have cloned and characterized a C5aR in rainbow trout, a teleost fish. The isolated cDNA encoded a 350-aa protein that showed the highest sequence similarity to C5aR from other species. Genomic analysis revealed the presence of one continuous exon encoding the entire open reading frame. Northern blot analysis showed significant expression of the trout C5a receptor (TC5aR) message in PBLs and kidney. Flow cytometric analysis showed that two Abs generated against two different areas of the extracellular N-terminal region of TC5aR positively stained the same leukocyte populations from PBLs. B lymphocytes and granulocytes comprised the majority of cells recognized by the anti-TC5aR. More importantly, these Abs inhibited chemotaxis of PBLs toward a chemoattractant fraction purified from complement-activated trout serum. Our data suggest that the split between C5aR and C3aR from a common ancestral molecule occurred before the emergence of teleost fish. Moreover, we demonstrate that the overall structure of C5aR as well as its role in chemotaxis have remained conserved for >300 million years.

Journal of Immunology↗
USGS2006

Evolution of the CD4 family: teleost fish possess two divergent forms of CD4 in addition to lymphocyte activation gene-3

The T cell coreceptor CD4 is a transmembrane glycoprotein belonging to the Ig superfamily and is essential for cell-mediated immunity. Two different genes were identified in rainbow trout that resemble mammalian CD4. One (trout CD4) encodes four extracellular Ig domains reminiscent off mammalian CD4, whereas the other (CD4REL) codes for two Ig domains. Structural motifs within the amino acid sequences suggest that the two Ig domains of CD4REL duplicated to generate the four-domain molecule of CD4 and the related gene, lymphocyte activation gene-3. Here we present evidence that both of these molecules in trout are homologous to mammalian CD4 and that teleosts encode an additional CD4 family member, lymphocyte activation gene-3, which is a marker for activated T cells. The syntenic relationships of similar genes in other teleost and non-fish genomes provide evidence for the likely evolution of CD4-related molecules in vertebrates, with CD4REL likely representing the primordial form in fish. Expression of both CD4 genes is highest in the thymus and spleen, and mRNA expression of these genes is limited to surface IgM- lymphocytes, consistent with a role for T cell functionality. Finally, the intracellular regions of both CD4 and CD4REL possess the canonical CXC motif involved in the interaction off CD4 with p56LCK, implying that similar mechanisms for CD4 + T cell activation are present in all vertebrates. Our results therefore raise new questions about T cell development and functionality in lower vertebrates that cannot be answered by current mammalian models and, thus, is of fundamental importance for understanding the evolution of cell-mediated immunity in gnathosomes. Copyright ?? 2006 by The American Association of Immunologists, Inc.

Journal of Immunology↗
USGS2026

Distributions of host heterogeneity in susceptibility show signatures of pathogen geographic structure in an insect baculovirus

Segregated variation between populations is a fundamental evolutionary process leading to parasite specialization, yet the resulting impacts on infection heterogeneity within populations are theoretically and empirically understudied. We asked whether the distribution of host susceptibility to infection within populations carries the signatures of geographic structure from pathogen local adaptation, maladaptation, or generalism in a nuclear polyhedrosis virus that infects the Gulf Fritillary butterfly Dione vanillae . For this virus, there is genetic support for two geographically distinct groups within San Diego County, based on whole genome sequencing of 16 virus isolates. Reciprocal laboratory infections showed evidence of two contrasting viral life history strategies: a ‘generalist’ phenotype that consistently infected variable hosts and a ‘specialist’ that performed slightly better in its local host population. As predicted by our theoretical model, the more consistent infection displayed by the generalist across populations corresponded to lower heterogeneity in susceptibility within populations, modeled as gamma distribution. Furthermore, the generalist phenotype was collected over a wider geographic range despite having a tenfold-lower mean infection rate than the specialist, suggesting that a strategy of more consistent infection provides key fitness advantages across diverse host populations. Intriguingly, when there is variation in host susceptibility, interpretations of pathogen local adaptation are dose-dependent. Measuring infectivity across multiple doses enables estimation of the whole distribution of susceptibility, which provides more reliable identification of pathogen specialization to its local host. Our work demonstrates how trait distributions and not only their mean values can carry quantifiable signatures of eco-evolutionary processes in interspecific interactions.

BioRxiv↗
USGS2000

Characterization of poxviruses from forest birds in Hawaii

Two strains of avian pox viruses were isolated from cutaneous lesions in Hawaiian crows ( Corvus hawaiiensis ) examined in 1994 and a third from a biopsy obtained in 1992 from an infected bird of the Apapane species ( Himatione sanguinea ) by inoculation of the chorioallantoic membranes (CAM) of developing chicken embryos. The resulting proliferative CAM lesions contained eosinophilic cytoplasmic inclusion bodies characteristic of pox virus infection. The pathogenicity of these three viruses in domestic chickens was mild as evidenced by the development of relatively minor lesions of short duration at the sites of inoculation. Their virulence in this host was similar to that of a fowlpox virus (FPV) vaccine strain and contrasted greatly with the ability of two field strains of FPV to produce extensive proliferative lesions. One of the Hawaiian crow pox virus isolates as well as the one originating from the Apapane species could be propagated in two secondary avian cell lines, QT-35 and LMH. A comparison of the restriction fragment length polymorphisms (RFLP) of the genomes of the two cell line-adapted viruses, generated by Eco RI digestion, revealed a limited degree of similarity. Moreover, neither profile was comparable to those of the two field isolates of FPV, which were almost indistinguishable from each other. Thus, based on the genetic distinctness of the two Hawaiian bird viruses, they appear to represent different strains of avipoxvirus.

Hawaii↗
USGS2005

Molecular identification of a papilloma virus from cutaneous lesions of captive and free-ranging Florida manatees

Cutaneous papillomatous lesions were biopsied from three captive Florida manatees ( Trichechus manatus latirostris ) at Homosassa Springs State Wildlife Park (HSSWP), Homosassa, Florida, USA, and from six free-ranging Florida manatees from Crystal and Homosassa rivers, Florida. Total DNA extracted from these lesions was assayed for the presence of papilloma virus genomes using the polymerase chain reaction (PCR) with primers that target the L1 capsid protein gene. The amplification generated DNA fragments 458 base pairs in length that encompassed a highly conserved domain within the L1 capsid protein and translated into identical polypeptides of 152 amino acids, suggesting the involvement of a single papilloma virus genotype. Multiple amino acid sequence and phylogenetic analyses of the L1 fragment indicated that the Florida manatee papilloma virus is a unique and quite distinct papillomavirus from other known papilloma viruses. The emergence of this new pathogen raises concerns about its potential impact on the already endangered Florida manatee.

Journal of Wildlife Diseases↗
USGS2008

Using amplified fragment length polymorphism analysis to differentiate isolates of Pasteurella multocida serotype 1

Avian cholera, an infectious disease caused by the bacterium Pasteurella multocida , kills thousands of North American wild waterfowl annually. Pasteurella multocida serotype 1 isolates cultured during a laboratory challenge study of Mallards ( Anas platyrhynchos ) and collected from wild birds and environmental samples during avian cholera outbreaks were characterized using amplified fragment length polymorphism (AFLP) analysis, a whole-genome DNA fingerprinting technique. Comparison of the AFLP profiles of 53 isolates from the laboratory challenge demonstrated that P. multocida underwent genetic changes during a 3-mo period. Analysis of 120 P. multocida serotype 1 isolates collected from wild birds and environmental samples revealed that isolates were distinguishable from one another based on regional and temporal genetic characteristics. Thus, AFLP analysis had the ability to distinguish P. multocida isolates of the same serotype by detecting spatiotemporal genetic changes and provides a tool to advance the study of avian cholera epidemiology. Further application of AFLP technology to the examination of wild bird avian cholera outbreaks may facilitate more effective management of this disease by providing the potential to investigate correlations between virulence and P. multocida genotypes, to identify affiliations between bird species and bacterial genotypes, and to elucidate the role of specific bird species in disease transmission.

California, Iowa, Missouri, Nebraska, Utah, Wiscon↗
USGS2021

Poecivirus is present in individuals with beak deformities in seven species of North American birds

Avian keratin disorder (AKD), a disease of unknown etiology characterized by debilitating beak overgrowth, has increasingly affected wild bird populations since the 1990s. A novel picornavirus, poecivirus, is closely correlated with disease status in Black-capped Chickadees ( Poecile atricapillus ) in Alaska. However, our knowledge of the relationship between poecivirus and beak deformities in other species and other geographic areas remains limited. The growing geographic scope and number of species affected by AKD-like beak deformities require a better understanding of the causative agent to evaluate the population-level impacts of this epizootic. Here, we tested eight individuals from six avian species with AKD-consistent deformities for the presence of poecivirus: Mew Gull ( Larus canus ), Hairy Woodpecker ( Picoides villosus ), Black-billed Magpie ( Pica hudsonia ), American Crow ( Corvus brachyrhynchos ), Red-breasted Nuthatch ( Sitta canadensis ), and Blackpoll Warbler ( Setophaga striata ). The birds were sampled in Alaska and Maine (1999−2016). We used targeted PCR followed by Sanger sequencing to test for the presence of poecivirus in each specimen and to obtain viral genome sequence from virus-positive host individuals. We detected poecivirus in all individuals tested, but not in negative controls (water and tissue samples). Furthermore, we used unbiased metagenomic sequencing to test for the presence of other pathogens in six of these specimens (Hairy Woodpecker, two American Crows, two Red-breasted Nuthatches, Blackpoll Warbler). This analysis yielded additional viral sequences from several specimens, including the complete coding region of poecivirus from one Red-breasted Nuthatch, which we confirmed via targeted PCR followed by Sanger sequencing. This study demonstrates that poecivirus is present in individuals with AKD-consistent deformities from six avian species other than Black-capped Chickadee. While further investigation will be required to explore whether there exists a causal link between this virus and AKD, this study demonstrates that poecivirus is not geographically restricted to Alaska, but rather occurs elsewhere in North America.

Alaska, Maine↗
USGS2022

Morbidity and mortality of Hawaiin geese (Branta sandvicensis) and Laysan albatross (Phoebastria immutabilis) associated with reticuloendotheliosis virus

Only one virus, Avipox, has been documented previously in wild birds in Hawaii. Using immunohistochemistry and PCR, we found that two native threatened Hawaiian Geese ( Branta sandvicensis ), one with multicentric histiocytoma and the other with toxoplasmosis, and one Laysan Albatross ( Phoebastria immutabilis ) with avian pox were infected with reticuloendotheliosis virus (REV). The virus was isolated from one of the geese by cell culture. Surveys of other Hawaiian geese with various pathologies, avian pox cases, and pox viral isolates using PCR failed to reveal REV, suggesting that the virus is uncommon, at least in samples examined. The full genome of the Gag, Pol, and Env genes were sequenced for all three infected birds and revealed geographic divergence of the Pol gene, suggesting it to be under strong selective pressure. Our finding of REV in Hawaii makes this only the second virus documented in native Hawaiian birds associated with pathology. Moreover, the presence of REV in a pelagic seabird is unusual. Future surveys should seek the reservoir of the virus in efforts to trace its origins.

Journal of Wildlife Diseases↗
USGS1991

Mitochondrial DNA in wildlife forensic science: Species identification of tissues

A common problem in wildlife law enforcement is identifying the species of origin of carcasses, meat, or blood when morphological characters such as hair or bones are not available. Immunological and protein electrophoretic (allozyme or general protein) procedures have been used in species identification with considerable success (Bunch et al. 1976, McClymont et al. 1982, Wolfe 1983, Mardini 1984, Pex and Wolfe 1985, Dratch 1986), However, immunological tests often are not sensitive enough to distinguish closely related species. Furthermore, electrophoretically detectable protein polymorphisms may be lacking in certain populations or species and may not be species-specific. Analysis of DNA in human and wildlife forensics has been shown to be a potentially powerful tool for identification of individuals (Jeffreys et al. 1985, Vassartet al. 1987, Thommasen et al. 1989). Differences in copy number and nucleotide sequence of repetitive sequences in the nuclear (chromosomal) DNA result in hypervariability and individual-specific patterns which have been termed DNA "fingerprints." However, these patterns may be too variable for species identification necessitating analyses of more conservative parts of the genome. Mitochondrial DNA (mtDNA) is haploid, maternally inherited, similar in nucleotide sequence among conspecifics from the same geographic region, and more suitable for species identification, in contrast to hypervariable DNA fingerprints. MtDNA has several characteristics which make it useful as a species-specific marker. In mammals, individuals have a single mtDNA genotype shared by all tissues. Because mtDNA is haploid and reflects only maternal ancestry, the mtDNA gene number in a population is 4 times less than the nuclear gene number (Birky et al. 1983). This can result in relatively rapid loss or fixation of mtDNA genotypes so that all individuals in a population may be descended from a single ancestral female in as few as 4N ( N = population size) generations (Avise et al., 1984). This suggests there is great potential for identification of species-specific or population-specific mtDNA genotypes, Within species, mtDNA genotypes are thought to be selectively neutral, i.e., there is little evidence that genotypes differ in fitness (Nigro and Prout 1990).

Wildlife Society Bulletin↗
USGS1992

Establishing paternity in Whooping Cranes ( Grus americana ) by DNA analysis

DNA fingerprinting was used to study paternity and genetic variability within a captive flock of Whooping Cranes ( Grus americana ). Fingerprint patterns for 42 individuals were obtained by digesting genomic crane DNAs with Hae III followed by electrophoresis, blotting, and hybridization to the M13 minisatellite probe. Despite finding reduced levels of genetic variation in the Whooping Crane due to a population "bottleneck," these polymorphisms were successfully used to determine paternity in six of seven cases of captive propagation where the maternal-offspring relationship was known, but where the sire was unknown. These determinations of paternity are required for effective genetic management of the crane flock. These results also revealed a number of heterozygous minisatellite loci that will be valuable in future assessments of genetic variability in this endangered species.

The Auk↗
USGS2005

Development of a subunit vaccine for infectious pancreatic necrosis virus using a baculovirus insect/larvae system

Various attempts to develop a vaccine against infectious pancreatic necrosis virus (IPNV) have not yielded consistent results. Thus, at present, no commercial vaccine is available that can be used with confidence to immunize fry of salmon and trout. We generated a cDNA clone of the large genome segment A of an IPNV Sp strain and expressed all structural protein genes in insect cells and larvae using a baculovirus expression system. Green fluorescent protein was also co-expressed as a reporter molecule. High yields of IPNV proteins were obtained and the structural proteins self assembled to form virus-like particles (VLPs). We tested the immunogenicity of the putative VLP antigen in immersion vaccine experiments (two concentrations) in rainbow trout (Oncorhynchus mykiss) fry, and by intraperitoneal immunisation of Atlantic salmon (Salmo salar) pre-smolts using an oil adjuvant formulation. Rainbow trout were challenged by immersion using either the Sp or the VR-299 strain of IPNV two or three weeks post-vaccination, while Atlantic salmon were bath challenged with Sp strain after two months, after parr-smolt transformation. In the rainbow trout fry challenged two weeks post-immunization, cumulative mortality rates three weeks post challenge were 14 % in the fry that had received the highest dose versus 8 % in the control groups. No indication of protection was seen in repeated trials using a lower dose of antigen and challenge three weeks post-immunisation. The cumulative mortality rate of intraperitoneally immunised Atlantic salmon post-smolts four weeks post challenge was lower (56 %) than in the control fish (77 %), showing a dose-response pattern.

Conference Paper↗
USGS2014

Molecular epidemiology and evolution of fish Novirhabdoviruses

The genus Novirhabdoviridae contains several of the important rhabdoviruses that infect fish hosts. There are four established virus species: Infectious hematopoietic necrosis virus (IHNV), Viral hemorrhagic septicemia virus (VHSV), Hirame rhabdovirus (HIRRV), and Snakehead rhabdovirus (SHRV). Viruses of these species vary in host and geographic range, and they have all been studied at the molecular and genomic level. As globally significant pathogens of cultured fish, IHNV and VHSV have been particularly well studied in terms of molecular epidemiology and evolution. Phylogenic analyses of hundreds of field isolates have defined five major genogroups of IHNV and four major genotypes of VHSV worldwide. These phylogenies are informed by the known histories of IHNV and VHSV, each involving a series of viral emergence events that are sometimes associated with host switches, most often into cultured rainbow trout. In general, IHNV has relatively low genetic diversity and a narrow host range, and has been spread from its endemic source in North American to Europe and Asia due to aquaculture activities. In contrast, VHSV has broad host range and high genetic diversity, and the source of emergence events is virus in widespread marine fish reservoirs in the northern Atlantic and Pacific Oceans. Common mechanisms of emergence and host switch events include use of raw feed, proximity to wild fish reservoirs of virus, and geographic translocations of virus or naive fish hosts associated with aquaculture.

Book chapter↗
USGS2012

Selecting sagebrush seed sources for restoration in a variable climate: ecophysiological variation among genotypes

Big sagebrush ( Artemisia tridentata ) communities dominate a large fraction of the United States and provide critical habitat for a number of wildlife species of concern. Loss of big sagebrush due to fire followed by poor restoration success continues to reduce ecological potential of this ecosystem type, particularly in the Great Basin. Choice of appropriate seed sources for restoration efforts is currently unguided due to knowledge gaps on genetic variation and local adaptation as they relate to a changing landscape. We are assessing ecophysiological responses of big sagebrush to climate variation, comparing plants that germinated from ~20 geographically distinct populations of each of the three subspecies of big sagebrush. Seedlings were previously planted into common gardens by US Forest Service collaborators Drs. B. Richardson and N. Shaw, (USFS Rocky Mountain Research Station, Provo, Utah and Boise, Idaho) as part of the Great Basin Native Plant Selection and Increase Project. Seed sources spanned all states in the conterminous Western United States. Germination, establishment, growth and ecophysiological responses are being linked to genomics and foliar palatability. New information is being produced to aid choice of appropriate seed sources by Bureau of Land Management and USFS field offices when they are planning seed acquisitions for emergency post-fire rehabilitation projects while considering climate variability and wildlife needs.

Report↗
USGS2010

Bison conservation initiative: Bison conservation genetics workshop: Report and recommendations

One of the first outcomes of the Department of the Interior (DOI) Bison Conservation Initiative was the Bison Conservation Genetics Workshop held in Nebraska in September 2008. The workshop brought together scientists from government agencies and non-governmental organizations with professional population geneticists to develop guidance for the genetic management of the federal bison herds. The scientists agreed on the basic tenets of genetic management for the DOI herds and discussed different approaches to meeting those goals. First, the 12 DOI herds are an irreplaceable resource for the long-term conservation of North American plains bison. Most of the herds show low levels of cattle introgression dating from the time when they were saved from extirpation; those herds should not be mixed without careful consideration as to their origin. Herds that show no evidence of cattle ancestry by the current molecular methods are the highest priority for protection from genetic mixing with any other bison herds. Second, despite the fact that most of the herds now managed by the U.S. government were founded with very few bison and have been maintained for many generations at relatively low population sizes, they do not show obvious effects of inbreeding. They have retained significant amounts of genetic variation by the standard measures, heterozygosity and allelic diversity. This may be explained in part by the fact that most of these herds are not remnants of a single population. Third, to preserve genetic variation in federal bison herds over decades and centuries, herds should be managed at a population or metapopulation level of 1,000 animals or more, with a sex ratio that enables competition between breeding bulls. The parks and refuges that currently have bison herds, with the exception of Yellowstone National Park, do not have enough land to support a population of this size. In the short term, it will be important to develop satellite herds to attain population targets, and develop a metapopulation structure between herds. Fourth and finally, the current methods used to evaluate the DOI bison herds, using mitochondrial DNA and a suite of nuclear DNA microsatellites, are highly informative at the herd level. They have confirmed relatedness of herds that we know from historical records have a common origin. They have detected cattle ancestry in most of the herds where it was suspected and have shown some loss of rare alleles. However, they do not sample across the bison genome, and the use of neutral genetic markers as the basis for selection of individual bison—either to breed or move to other herds—would be better supported by more high-resolution molecular methods currently under development.

Natural Resource Report↗
USGS2015

Hyla chrysoscelis (Cope’s gray treefrog) x Hyla cinerea (green treefrog): putative natural hybrid

Naturally–occurring hybrid treefrogs have been occasionally found in the eastern United States. However, these hybrids are almost always between members of the same species group. On 10 Jun 2014, at 2145 h, we located an individual making an unusual advertisement call along Bayou Manual Road in Sherburne Wildlife Management Area in the Atchafalaya Basin of south-central Louisiana, USA, and brought it back to the laboratory for further study. Physically, the treefrog appeared intermediate between a Green Treefrog and a Cope’s Gray Treefrog, which are members of different species groups. Call analysis also showed the individual to be intermediate between the two putative parental species. Flow cytometry was used to estimate the total genome size from nuclei of whole blood cells, and also determined the individual to be intermediate of the putative parental species. Despite vocalizing for mates, the hybrid did not appear to have viable spermatozoa, and was likely the result of an anomalous mis-mating event between a male Cope’s Gray Treefrog and a female Green Treefrog. To our knowledge, natural hybrids between a Cope’s Gray Treefrog and a Green Treefrog have not been previously reported.

Louisiana↗
USGS1989

Response of salmonid fish to artificial infection with chum salmon virus

In the fall of 1978, a reovirus was isolated from normal-appearing adult chum salmon (Oncorhynchus keta ) returning to the Tokushibetsu Hatchery in Hokkaido, Japan (Winton et al 1981). The chum salmon virus (CSW) was recovered in the chinook salmon ( Oncorhynchus tshawytscha ) embryo cell line (CHSE-214) where it replicated at 15-20 C, producing foci of syncytia in the monolayer. Electron microscopy revealed icosahedral particles, 75 nm in diameter, with a double capsid. The virus was not inactivated by chloroform or inhibited by fluorodeoxyuridine. It was unstable at 56 C, did not hemagglutinate human type 0 erythrocytes, and had a density of 1.33 g/ml in CsCl . The virus was not neutralized by antiserum against infectious pancreatic necrosis virus or mammalian reovirus serotypes 1, 2 or 3 (Winton 1981). Electrophoretic analysis showed the genome was composed of three large, three medium, and five small segments of double stranded RNA (dsRNA) that ranged from 0.37-2.5 x 10% molecular weight. The virions contained five major structural proteins and several minor proteins (Winton et al 1983).

Book chapter↗
USGS1989

Isolation of a reovirus from coho salmon ( Oncorhynchus kisutch ) in Oregon, USA

Reoviruses isolated from aquatic animals share certain common characteristics: (1) a typical reovirus-like morphology which shows an icosahedral particle with a double capsid that is approximately 75 nm in diameter; (2) a genome with eleven segments of double-stranded RNA (dsRNA) distributed as three large, three medium and five small segments with a total molecular weight of approximately 15 x 106; (3) a virion composed of five major and several minor structural proteins that range in molecular weight from 32,000 to 137,000; and (4) form plaque-like syncytia in monolayer cultures of fish cells. Intact virus particles have buoyant densities in CsCl of 1.34 to 1.36 g/ml. The viruses have been isolated from fish and shellfish collected in both the marine and freshwater environments and will replicate in several fish cell lines (Plumb et al., 1979; Meyers and Hirai, 1980; Winton et al., 1981; Nagabayashi and Mori, 1983; Hedrick et al., 1984; Chen and Jiang, 1984). The original four aquatic reovirus isolates have been compared by Winton et al., 1987.

Book chapter↗
USGS1998

A ribonuclease protection assay can distinguish spring viremia of carp virus from pike fry rhabdovirus

Thirteen rhabdovirus isolates from 10 teleost fish species as well as reference strains of spring viraemia of carp virus (SVCV) and pike fry rhabdovirus (PFRV) cross-reacted in an indirect immunofluorescence assay and were thus indistinguishable by this method. A ribonuclease protection assay (RPA) using a super(32)P-labeled RNA probe made from a cloned copy of the full length SVCV glycoprotein (G) gene was able to discriminate clearly between the type strains of SVCV and PFRV and among the 13 rhabdovirus isolates. Results for the RPA were generally in agreement with standard serum neutralisation assays; however, the RPA was also able to detect genomic differences between isolates of SVCV. These results have implications for fish disease control programs for SVCV.

Bulletin of the European Association of Fish Patho↗