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USGS2014

Biodiversity loss and infectious diseases

When conservation biologists think about infectious diseases, their thoughts are mostly negative. Infectious diseases have been associated with the extinction and endangerment of some species, though this is rare, and other factors like habitat loss and poorly regulated harvest still are the overwhelming drivers of endangerment. Parasites are pervasive and play important roles as natural enemies on par with top predators, from regulating population abundances to maintaining species diversity. Sometimes, parasites themselves can be endangered. However, it seems unlikely that humans will miss extinct parasites. Parasites are often sensitive to habitat loss and degradation, making them positive indicators of ecosystem “health”. Conservation biologists need to carefully consider infectious diseases when planning conservation actions. This can include minimizing the movement of domestic and invasive species, vaccination, and culling.

Book chapter↗

USGS2010

A reverse genetics system for the Great Lakes strain of viral hemorrhagic septicemia virus: the NV gene is required for pathogenicity

Viral hemorrhagic septicemia virus (VHSV), belonging to the genus Novirhabdovirus in the family of Rhabdoviridae , causes a highly contagious disease of fresh and saltwater fish worldwide. Recently, a novel genotype of VHSV, designated IVb, has invaded the Great Lakes in North America, causing large-scale epidemics in wild fish. An efficient reverse genetics system was developed to generate a recombinant VHSV of genotype IVb from cloned cDNA. The recombinant VHSV (rVHSV) was comparable to the parental wild-type strain both in vitro and in vivo, causing high mortality in yellow perch ( Perca flavescens ). A modified recombinant VHSV was generated in which the NV gene was substituted with an enhanced green fluorescent protein gene (rVHSV-ΔNV-EGFP), and another recombinant was made by inserting the EGFP gene into the full-length viral clone between the P and M genes (rVHSV-EGFP). The in vitro replication kinetics of rVHSV-EGFP was similar to rVHSV; however, the rVHSV-ΔNV-EGFP grew 2 logs lower. In yellow perch challenges, wtVHSV and rVHSV induced 82-100% cumulative per cent mortality (CPM), respectively, whereas rVHSV-EGFP produced 62% CPM and rVHSV-ΔNV-EGFP caused only 15% CPM. No reversion of mutation was detected in the recovered viruses and the recombinant viruses stably maintained the foreign gene after several passages. These results indicate that the NV gene of VHSV is not essential for viral replication in vitro and in vivo, but it plays an important role in viral replication efficiency and pathogenicity. This system will facilitate studies of VHSV replication, virulence, and production of viral vectored vaccines.

Marine Biotechnology↗

USGS2013

Has the time come for big science in wildlife health?

The consequences of wildlife emerging diseases are global and profound with increased burden on the public health system, negative impacts on the global economy, declines and extinctions of wildlife species, and subsequent loss of ecological integrity. Examples of health threats to wildlife include Batrachochytrium dendrobatidis , which causes a cutaneous fungal infection of amphibians and is linked to declines of amphibians globally; and the recently discovered Pseudogymnoascus (Geomyces) destructans , the etiologic agent of white nose syndrome which has caused precipitous declines of North American bat species. Of particular concern are the novel pathogens that have emerged as they are particularly devastating and challenging to manage. A big science approach to wildlife health research is needed if we are to make significant and enduring progress in managing these diseases. The advent of new analytical models and bench assays will provide us with the mathematical and molecular tools to identify and anticipate threats to wildlife, and understand the ecology and epidemiology of these diseases. Specifically, new molecular diagnostic techniques have opened up avenues for pathogen discovery, and the application of spatially referenced databases allows for risk assessments that can assist in targeting surveillance. Long-term, systematic collection of data for wildlife health and integration with other datasets is also essential. Multidisciplinary research programs should be expanded to increase our understanding of the drivers of emerging diseases and allow for the development of better disease prevention and management tools, such as vaccines. Finally, we need to create a National Fish and Wildlife Health Network that provides the operational framework (governance, policies, procedures, etc.) by which entities with a stake in wildlife health cooperate and collaborate to achieve optimal outcomes for human, animal, and ecosystem health.

EcoHealth↗

USGS2011

Deltamethrin flea-control preserves genetic variability of black-tailed prairie dogs during a plague outbreak

Genetic variability and structure of nine black-tailed prairie dog (BTPD, Cynomys ludovicianus ) colonies were estimated with 15 unlinked microsatellite markers. A plague epizootic occurred between the first and second years of sampling and our study colonies were nearly extirpated with the exception of three colonies in which prairie dog burrows were previously dusted with an insecticide, deltamethrin, used to control fleas (vectors of the causative agent of plague, Yersinia pestis ). This situation provided context to compare genetic variability and structure among dusted and non-dusted colonies pre-epizootic, and among the three dusted colonies pre- and post-epizootic. We found no statistical difference in population genetic structures between dusted and non-dusted colonies pre-epizootic. On dusted colonies, gene flow and recent migration rates increased from the first (pre-epizootic) year to the second (post-epizootic) year which suggested dusted colonies were acting as refugia for prairie dogs from surrounding colonies impacted by plague. Indeed, in the dusted colonies, estimated densities of adult prairie dogs (including dispersers), but not juveniles (non-dispersers), increased from the first year to the second year. In addition to preserving BTPDs and many species that depend on them, protecting colonies with deltamethrin or a plague vaccine could be an effective method to preserve genetic variability of prairie dogs.

Montana↗

USGS1985

Immune response of channel catfish, Ictalurus punctatus Rafinesque, to bacterial and protozoan antigens administered by three routes

Experiments were conducted to measure the agglutinating antibody response of channel catfish to a particulate and a soluble antigen administered simultaneously by one of three routes. Specific antibody production in response to administration of particulate brucella tube-test antigen via intramuscular injection, oral drench, and topical application peaked at 3 weeks in channel catfish held under identical conditions at 21°C. The humoral antibody response was greater in fish injected with antigen than in the orally or topically exposed channel catfish. Specific antibody was also produced by the same channel catfish in response to the administration of a soluble ciliary preparation derived from Tetrahymena pyriformis Lwoff. Intramuscular injection of Tetrahymena antigen was the only route that produced detectable levels of circulating antibody as measured by direct agglutination of live T. pyriformis cells. The response to the soluble antigen also peaked at 3 weeks. Neither antigen elicited detectable agglutinins in cutaneous mucus, regardless of the vaccination method used.

Aquaculture↗

USGS1991

Recent advances in detection and control of infectious hematopoietic necrosis virus in aquaculture

Infectious hematopoietic necrosis (IHN) is one of the most important viral diseases of salmon and trout reared in culture. The disease remains untreatable with avoidance being the only control measure. Much has been learned about the chemical, physical, and serological characteristics of the rhabdovirus causing IHN, but critical gaps exist in our understanding of the biology of the virus in nature. The tools of molecular biology have provided improved methods for detection of pathogens and new strategies for control of viral diseases. This paper reviews several recent improvements in methods for detecting infectious hematopoietic necrosis virus including the application of enzyme-linked immunosorbent assays, development of monoclonal antibodies and DNA probes, and use of the polymerase chain reaction. New strategies for control of IHN through the use of better water treatment, more resistant fish, antiviral drugs or chemicals, and new generation vaccines are discussed.

Annual Review of Fish Diseases↗

USGS2019

Health and disease treatment in captive and reintroduced Whooping Cranes

The principles and applications of medicine in Whooping Cranes ( Grus americana ) are similar to that for other crane species, though the necessity of managing Whooping Crane health across the captive, reintroduced, and remnant populations poses particular challenges. In this chapter, we review the important aspects of crane medicine relevant to managing Whooping Cranes in captive and reintroduced environments. We pay special attention to viral, parasitic, fungal, and bacterial diseases that are important to manage for reintroduction efforts to be successful. We also introduce some common aspects of treatment and surgery. Anesthesia is important for successful surgery and is discussed. Radio transmitter implant surgery, as it applies to studying causes of mortality in wild Whooping Crane chicks, is included as a subject of special interest to recovery efforts. Orthopedic problems, which pose a major challenge to the healthy development of young, captive-reared cranes, are discussed. Treatment and vaccination recommendations are made to aid in producing healthy Whooping Cranes for reintroduction to the wild. The literature about crane medicine is reviewed along with the personal insights of the three authors, and an overview of the development of crane medicine programs is included.

Book chapter↗

USGS2008

Fish Rhabdoviruses

Many important viral pathogens of fish are members of the family Rhabdoviridae . The viruses in this large group cause significant losses in populations of wild fish as well as among fish reared in aquaculture. Fish rhabdoviruses often have a wide host and geographic range, and infect aquatic animals in both freshwater and seawater. The fish rhabdoviruses comprise a diverse collection of isolates that can be placed in one of two quite different groups: isolates that are members of the established genus Novirhabdovirus , and those that are most similar to members of the genus Vesiculovirus . Because the diseases caused by fish rhabdoviruses are important to aquaculture, diagnostic methods for their detection and identification are well established. In addition to regulations designed to reduce the spread of fish viruses, a significant body of research has addressed methods for the control or prevention of diseases caused by fish rhabdoviruses, including vaccination. The number of reported fish rhabdoviruses continues to grow as a result of the expansion of aquaculture, the increase in global trade, the development of improved diagnostic methods, and heightened surveillance activities. Fish rhabdoviruses serve as useful components of model systems to study vertebrate virus disease, epidemiology, and immunology.

Book chapter↗

USGS2020

Genomic architecture and repertoire of the rainbow trout immunoglobulin light chain genes

The genomic loci encoding the four immunoglobulin light chains (IgL1, IgL2, IgL3, and IgL4) in the Swanson trout genome assembly were annotated in order to provide a measurement of the potential IgL repertoire. IgL1 and IgL3 gene segments are co-localized on chromosomes 21, 18, 15, and 7 while IgL2 and IgL4 were found on chromosomes 13 and 17, respectively. In total, 48 constant (C L ), 87 variable (V L ), and 59 joining (J L ) productive genes are described. Pairwise alignment of the V L segments revealed that they belong to nine different families, three of which (kappa IV, V, and VI) are described for the first time in this study. V L and C L sequences on chromosome 15 and 21 and those on chromosomes 7 and 18 clustered together in phylogenetic analysis. PCR was used to examine IgL C L and V L genes in 9 lines of rainbow trout. IgL4 in the Hot Creek and Golden trout lines was missing 42 nucleotides resulting in a loss of 14 amino acids. The sigma IV variable family was completely absent from the Swanson, Arlee, Hot Creek, and wild type lines and silenced in the Skamania line with the addition of 176 bp mini-satellite insert. Similarly, the Whale Rock, Arlee, and wild type lines were all found to encode two sigma II products, a functional 252 bp product and a larger 425 bp product that contained a 172 bp insert. Results from this study indicate that there are genomic differences in IgL repertoire between different lines of trout that could affect humoral immune responses post vaccination and during disease.

Developmental and Comparative Immunology↗

USGS2016

Temporal, geographic, and host distribution of avian paramyxovirus 1 (Newcastle disease virus)

Newcastle disease is caused by virulent forms of avian paramyxovirus of serotype 1 (APMV-1) and has global economic importance. The disease reached panzootic proportions within two decades after first being identified in 1926 in the United Kingdom and Indonesia and still remains endemic in many countries across the world. Here we review information on the host, temporal, and geographic distribution of APMV-1 genetic diversity based on the evolutionary systematics of the complete coding region of the fusion gene. Strains of APMV-1 are phylogenetically separated into two classes (class I and class II) and further classified into genotypes based on genetic differences. Class I viruses are genetically less diverse, generally present in wild waterfowl, and are of low virulence. Class II viruses are genetically and phenotypically more diverse, frequently isolated from poultry with occasional spillovers into wild birds, and exhibit a wider range of virulence. Waterfowl, cormorants, and pigeons are natural reservoirs of all APMV-1 pathotypes, except viscerotropic velogenic viruses for which natural reservoirs have not been identified. Genotypes I and II within class II include isolates of high and low virulence, the latter often being used as vaccines. Viruses of genotypes III and IX that emerged decades ago are now isolated rarely, but may be found in domestic and wild birds in China. Containing only virulent viruses and responsible for the majority of recent outbreaks in poultry and wild birds, viruses from genotypes V, VI, and VII, are highly mobile and have been isolated on different continents. Conversely, virulent viruses of genotypes XI (Madagascar), XIII (mainly Southwest Asia), XVI (North America) and XIV, XVII and XVIII (Africa) appear to have a more limited geographic distribution and have been isolated predominantly from poultry.

Infection, Genetics and Evolution↗

USGS2018

Broad-spectrum antiviral JL122 blocks infection and inhibits transmission of aquatic rhabdoviruses

The aquaculture industry is growing rapidly to meet the needs for global protein consumption. Viral diseases in aquaculture are quite challenging due to lack of treatment options as well as limited injection-delivery vaccines, which are costly. Thus, water-immersion antiviral treatments are highly desirable. This study focused on broad-spectrum, light-activated antivirals that target the viral membrane (envelope) of viruses to prevent viral-cell membrane fusion, ultimately blocking viral entry into cells. Of the tested small-molecules, JL122, a new broad-spectrum antiviral previously unexplored against aquatic viruses, blocked infection of three aquatic rhabdoviruses (IHNV, VHSV and SVCV) in cell culture and in two live fish challenge models. Importantly, JL122 inhibited transmission of IHNV from infected to uninfected rainbow trout. Further, the effective antiviral concentrations were not toxic to cells or susceptible fish. These results show promise for JL122 to become an immersion treatment option for outbreaks of aquatic enveloped viral infections.

Virology↗

USGS2016

Determining the 95% limit of detection for waterborne pathogen analyses from primary concentration to qPCR

The limit of detection (LOD) for qPCR-based analyses is not consistently defined or determined in studies on waterborne pathogens. Moreover, the LODs reported often reflect the qPCR assay alone rather than the entire sample process. Our objective was to develop an approach to determine the 95% LOD (lowest concentration at which 95% of positive samples are detected) for the entire process of waterborne pathogen detection. We began by spiking the lowest concentration that was consistently positive at the qPCR step (based on its standard curve) into each procedural step working backwards (i.e., extraction, secondary concentration, primary concentration), which established a concentration that was detectable following losses of the pathogen from processing. Using the fraction of positive replicates (n = 10) at this concentration, we selected and analyzed a second, and then third, concentration. If the fraction of positive replicates equaled 1 or 0 for two concentrations, we selected another. We calculated the LOD using probit analysis. To demonstrate our approach we determined the 95% LOD for Salmonella enterica serovar Typhimurium, adenovirus 41, and vaccine-derived poliovirus Sabin 3, which were 11, 12, and 6 genomic copies (gc) per reaction (rxn), respectively (equivalent to 1.3, 1.5, and 4.0 gc L −1 assuming the 1500 L tap-water sample volume prescribed in EPA Method 1615). This approach limited the number of analyses required and was amenable to testing multiple genetic targets simultaneously (i.e., spiking a single sample with multiple microorganisms). An LOD determined this way can facilitate study design, guide the number of required technical replicates, aid method evaluation, and inform data interpretation.

Water Research↗

USGS2000

Toxicity of acidified chitosan for cultured rainbow trout (Oncorhynchus mykiss)

Chitosan is a deacetylation product of chitin. It is used as a flocculent for sewage and brewery wastes, and as a chelator of heavy metals. In aquaculture, chitosan has been used as an immunostimulant for protection against bacterial diseases in fish, for controlled release of vaccines, and as a diet supplement. Chitosan has generally been considered to be nontoxic to animals, but when it was dissolved in acetic acid and added to a culture system at 1.0 ppm to remove organic solids, we found acute toxicity to rainbow trout ( Oncorhynchus mykiss ). In controlled experiments to determine the extent of toxicity, we found that trout died after several hours exposure to 0.75 ppm and died in 24 h after exposure to 0.075 ppm. Exposure to 0.038 ppm resulted in mortality after 6 days exposure, while exposure to 0.019 ppm resulted in no mortality after 14 days exposure. Histological examination of gills, skin, muscle, and internal organs indicated significant and consistent pathological changes only in gills. Lifting of lamellar epithelium, hypertrophy and hyperplasia of lamellar epithelial cells occurred in trout exposed to 0.019 and 0.038 ppm. In trout exposed to 0.75 or 0.075 ppm chitosan, large areas of lamellar fusion were observed. These results show that soluble acidified chitosan is highly toxic to rainbow trout even at low concentrations.

Aquaculture↗

USGS2003

Induction of lupus autoantibodies by adjuvants

Exposure to the hydrocarbon oil pristane induces lupus specific autoantibodies in non-autoimmune mice. We investigated whether the capacity to induce lupus-like autoimmunity is a unique property of pristane or is shared by other adjuvant oils. Seven groups of 3-month-old female BALB/cJ mice received a single intraperitoneal injection of pristane, squalene (used in the adjuvant MF59), incomplete Freund's adjuvant (IFA), three different medicinal mineral oils, or saline, respectively. Serum autoantibodies and peritoneal cytokine production were measured. In addition to pristane, the mineral oil Bayol F (IFA) and the endogenous hydrocarbon squalene both induced anti-nRNP/Sm and -Su autoantibodies (20% and 25% of mice, respectively). All of these hydrocarbons had prolonged effects on cytokine production by peritoneal APCs. However, high levels of IL-6, IL-12, and TNF?? production 2-3 months after intraperitoneal injection appeared to be associated with the ability to induce lupus autoantibodies. The ability to induce lupus autoantibodies is shared by several hydrocarbons and is not unique to pristane. It correlates with stimulation of the production of IL-12 and other cytokines, suggesting a relationship with a hydrocarbon's adjuvanticity. The potential to induce autoimmunity may complicate the use of oil adjuvants in human and veterinary vaccines. ?? 2003 Elsevier Ltd. All rights reserved.

Journal of Autoimmunity↗

USGS1991

Responses to in vitro and in vivo immunisations with Aeromonas salmonicida O antigen bacterins in rainbow trout (Oncorhynchus mykiss)

Immunological assays were developed to test for the effects of immunisation with Aeromonas salmonicida O antigen bacterin in rainbow trout ( Oncorhynchus mykiss ) in vitro and in vivo . Trout spleen sections were immunised in vitro by incubation in media containing 100, 10, and 1 μ g ml −1 of the A. salmonicida O antigen. After 10 days at 15°C, the individual sections were minced and cell suspensions assayed for the effects of the bacterin on non-specific and specific immune responses. Assays of the cell suspensions showed that the neutrophil oxidative activity, phagocytic capabilities, numbers of plaque-forming cells (PFC) and circulating antibody titres all increased in proportion to the bacterin dosage. Trout immunised in vivo with 100 μ g by injection and by 100 μ g ml −1 bath with the O antigen, showed similar responses to spleen sections immunised in vitro . The appearance of non-specific PFC against unlabelled sheep red blood cells was highest in spleens immunised in vitro and in the fish immunised by injection; non-specific PFC rarely occurred in O antigen bathed fish and in control fish. The development of these assays may enable biologists to follow the effects on the non-specific defence mechanisms and the specific immune response in cultivated fish populations after, immunisation with A. salmonicida bacterins and thus aid in the development of protective vaccination against furunculosis.

Fish and Shellfish Immunology↗

USGS2004

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

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

Environmental Biology of Fishes↗

USGS2026

Rapid expansion of genotype D1.1 A(H5N1) influenza viruses in wild birds across North America during the 2024 migratory season

In late 2021, high pathogenicity avian influenza A(H5N1) clade 2.3.4.4b viruses entered North America and reassorted rapidly with local avian influenza viruses. In September 2024, we detected a new reassortant later classified as genotype D1.1. Using active and passive avian influenza surveillance across Canada and the USA, we tracked the emergence and rapid spread of D1.1 viruses in wild birds during the 2024 fall migration. Phylodynamic analysis showed that D1.1 viruses formed a monophyletic group and displaced earlier A(H5) genotypes across several flyways. Their expansion coincided with detections in other hosts, including 17 human cases, 4 of which were severe or fatal. None of the mammalian-adaptive markers detected in human cases were found in wild bird viruses, and candidate vaccine viruses retained antigenic cross-reactivity with D1.1 strains.

Nature medicine↗

USGS2026

Loma salmonae and related species

Loma salmonae is a microsporidium that infects Pacific salmon and causes a gill inflammatory syndrome known as microsporidial gill disease of salmon. This disease has been mostly associated with netpen-farmed Chinook salmon ( Oncorhynchus tshawytscha ) in British Columbia, Canada. Clinical, diagnostic, pathological aspects of disease, as well as approaches for disease avoidance in salmon aquaculture are discussed. A laboratory infection model in rainbow trout was used to determine life cycle-stages, transmission dynamics, pathophysiology, influence of temperature, and to test therapeutics applicable to aquaculture. This experimental model has been informative on various approaches of disease control, including the development of a promising vaccine. In addition to improving fish health in salmon farming in North America, this L. salmonae model will be applicable to other microsporidial diseases that may be encountered in emerging aquaculture regions.

Book chapter↗