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At least 703 records · Page 39Linked to original sources
USGS2017

A framework for modeling emerging diseases to inform management

The rapid emergence and reemergence of zoonotic diseases requires the ability to rapidly evaluate and implement optimal management decisions. Actions to control or mitigate the effects of emerging pathogens are commonly delayed because of uncertainty in the estimates and the predicted outcomes of the control tactics. The development of models that describe the best-known information regarding the disease system at the early stages of disease emergence is an essential step for optimal decision-making. Models can predict the potential effects of the pathogen, provide guidance for assessing the likelihood of success of different proposed management actions, quantify the uncertainty surrounding the choice of the optimal decision, and highlight critical areas for immediate research. We demonstrate how to develop models that can be used as a part of a decision-making framework to determine the likelihood of success of different management actions given current knowledge.

Emerging Infectious Diseases↗
USGS2017

Reassortment of influenza A viruses in wild birds in Alaska before H5 Clade 2.3.4.4 Outbreaks

Sampling of mallards in Alaska during September 2014–April 2015 identified low pathogenic avian influenza A virus (subtypes H5N2 and H1N1) that shared ancestry with highly pathogenic reassortant H5N2 and H5N1 viruses. Molecular dating indicated reassortment soon after interhemispheric movement of H5N8 clade 2.3.4.4, suggesting genetic exchange in Alaska or surrounds before outbreaks.

Emerging Infectious Diseases↗
USGS1994

Association of viral hemorrhagic septicemia virus with epizootic hemorrhages of the skin in Pacific herring Clupea harengus pallasi from Prince William Sound and Kodiak Island, Alaska, USA

Only one-third of the Pacific hernng Clupea harengus pallasi expected to spawn in Prince William Sound (PWS), Alaska, USA, in sprlng 1993 were observed. Of these herring, 15 to 43 '% had external ulcers or subdermal hemorrhages of the skln and fins. A rhabdovirus identified as the North American strain of viral hemorrhagic septicemia virus (VHSV) was isolated from affected herring and 1 Pacific cod Gadus macrocephalus with skin lesions from PWS, and from herring with similar lesions collected near Kodiak Island. No other pathogens were detected in the herring examined. Although VHSV may have been responsible for the skin lesions, there was no confirmed mass herring mortality observed in PWS; hence the actual cause of the reduced herring numbers is still unknown. The same strain of VHSV was subsequently isolated from captive juvenile herring collected from Auke Bay, Alaska, near Juneau, from herring in British Columbia, Canada, and from Puget Sound, Washington, USA. These findings suggest the virus is an opportunistic pathogen that is widely indigenous to Pacific herring populations in the Pacific Northwest and that herring are a significant marine reservoir for North American VHSV.

Alaska↗
USGS2008

Widespread occurrence of the amphibian chytrid fungus Batrachochytrium dendrobatidis in the southeastern USA

From 1999 to 2006, we sampled >1200 amphibians for the fungal pathogen Batrachochytnum dendrobatidis (Bd) at 30 sites in the southeastern USA. Using histological techniques or PCR assays, we detected chytrid infection in 10 species of aquatic-breeding amphibians in 6 states. The prevalence of chytrid infection was 17.8% for samples of postmetamorphic amphibians examined using skin swab-PCR assays (n = 202 samples from 12 species at 4 sites). In this subset of samples, anurans had a much higher prevalence of infection than caudates (39.2% vs. 5.5%, respectively). Mean prevalence in ranid frogs was 40.7 %. The only infected salamanders were Notophthalmus viridescens at 3 sites. We found infected amphibians from late winter through late spring and in 1 autumn sample. Although we encountered moribund or dead amphibians at 9 sites, most mortality events were not attributed to Bd. Chytridiomycosis was established as the probable cause of illness or death in fewer than 10 individuals. Our observations suggest a pattern of widespread and subclinical infections. However, because most of the sites in our study were visited only once, we cannot dismiss the possibility that chytridiomycosis is adversely affecting some populations. Furthermore, although there is no evidence of chytrid-associated declines in our region, the presence of this pathogen is cause for concern given global climate change and other stressors. Although presence-absence surveys may still be needed for some taxa, such as bufonids, we recommend that future researchers focus on potential population-level effects at sites where Bd is now known to occur. ?? Inter-Research 2008.

Southeast↗
USGS2010

Epizootic ulcerative syndrome caused by Aphanomyces invadans in captive bullseye snakehead Channa marulius collected from south Florida, USA

Epizootic ulcerative syndrome (EUS) caused by the oomycete Aphanomyces invadans is an invasive, opportunistic disease of both freshwater and estuarine fishes. Originally documented as the cause of mycotic granulomatosis of ornamental fishes in Japan and as the cause of EUS of fishes in southeast Asia and Australia, this pathogen is also present in estuaries and freshwater bodies of the Atlantic and gulf coasts of the USA. We describe a mass mortality event of 343 captive juvenile bullseye snakehead Channa marulius collected from freshwater canals in Miami-Dade County, Florida. Clinical signs appeared within the first 2 d of captivity and included petechiae, ulceration, erratic swimming, and inappetence. Histological examination revealed hyphae invading from the skin lesions deep into the musculature and internal organs. Species identification was confirmed using a species-specific PCR assay. Despite therapeutic attempts, 100% mortality occurred. This represents the first documented case of EUS in bullseye snakehead fish collected from waters in the USA. Future investigation of the distribution and prevalence of A. invadans within the bullseye snakehead range in south Florida may give insight into this pathogen-host system.

Florida↗
USGS2020

Amphibian chytrid prevalence on boreal toads in SE Alaska and NW British Columbia: Tests of habitat, life stages, and temporal trends

Tracking and understanding variation in pathogens such as Batrachochytrium dendrobatidis ([Bd]), which causes amphibian chytridiomycosis and has caused population declines globally, is a priority for many land managers. However, there has been relatively little sampling of amphibian communities at high latitudes. We used skin swabs collected during 2005–2017 from boreal toads (Anaxyrus boreas; N = 248), in southeast Alaska (USA; primarily in Klondike Gold Rush National Historical Park [KLGO]) and northwest British Columbia (Canada) to determine how Bd prevalence varied across life stages, habitat characteristics, local species richness, and time. Across all years, Bd prevalence peaked in June and was >3 times greater for adult toads (37.5%) vs. juveniles and metamorphs (11.2%). Bd prevalence for toads in the KLGO area, where other amphibian species are rare or absent, was highest from river habitats (55.0%), followed by human-modified upland wetlands (32.3%) and natural upland wetlands (12.7%) — the same rank-order these habitats are used for toad breeding. No Columbia spotted frogs (N = 12) or wood frogs (N = 2) from the study area tested Bd-positive, although all were from an area of low host density where Bd has not been detected. Prevalence of Bd on toads in the KLGO area decreased during 2005–2015. This trend from a largely single-species system may be encouraging or concerning, depending on how Bd is affecting vital rates, and emphasizes the need to understand effects of pathogens before translating disease prevalence into management actions.

Southeastern Alaska, Northwestern British Columbia↗
USGS1999

Pathogenecity of Ichthyophonus hoferi for laboratory-reared Pacific herring ( Clupea pallasi ) and its early appearance in wild Puget Sound herring

Laboratory-reared pathogen-free Pacific herring were exposed to pure cultures of Ichthyophonus hoferi, and reproduced the disease seen in naturally infected fish--thus fulfilling Koch's Postulates. Pathogen-free herring used in this study were reared from artificially spawned eggs incubated in filtered, UV-sterilized seawater, eliminating the variables associated with multiple infections, which are common in wild herring. Wild free-ranging herring were captured monthly from June through October by dip net from 'herring balls' located in the northern Puget Sound. I. hoferi infections were identified in these fish soon after metamorphoses, about 4 mo post-hatch. The prevalence increased from 5 to 6% in 0-yr fish to 24% in 1-yr-old fish to 50 to 70% in fish over 2 yr old, with no associated increase in mortality. The route of natural transmission to wild herring was not determined, but carnivorous fish became infected and died when they were experimentally fed tissues infected with the organism. In vitro culture of tissues was the most sensitive method for identifying both clinical and subclinical infections.

Washington↗
USGS2018

Waterfowl spring migratory behavior and avian influenza transmission risk in the changing landscape of the East Asian-Australasian Flyway

Avian influenza has advanced from a regional concern to a global health issue with significant economic, trade, and public health implications. Wild birds, particularly waterfowl (Anseriformes), are known reservoirs for low-pathogenic avian influenza viruses (AIV) and recent studies have shown their potential in the spread of highly pathogenic forms of virus. East Asia remains an epicenter for the emergence of novel strains of AIV, however, information on movement ecology of waterfowl, and subsequently a clearer understanding of disease transmission risks in this region has been greatly lacking. To address this, we marked two species of wild waterfowl, northern pintail (Anas acuta) and Eurasian wigeon (Anas penelope) , with satellite transmitters on their wintering grounds in Hong Kong, China to study the northward spring migration in the East Asian-Australasian Flyway in relation to disease transmission factors. Northern pintail were found to initiate migration 42 days earlier, travel 2,150 km farther, and perform 4.4 more stopovers than Eurasian wigeon. We found both species used similar stopover locations including areas along the Yangtze River near Shanghai, Bohai Bay and Korea Bay in rapidly developing regions of the Yellow Sea, and the Sea of Okhotsk where the species appeared to funnel through a migratory bottleneck. Both species appeared to exhibit strong habitat selection for rice paddies during migration stopovers, a habitat preference which has the potential to influence risks of AIV outbreaks as rapid land use and land cover changes occur throughout China. Both species had greatest association with H5N1 outbreaks during the early stages of migration when they were at lower latitudes. While Eurasian wigeon were not associated with outbreaks after the mean date of wintering ground departures, northern pintail were associated with outbreaks until the majority of individuals departed from the Yellow Sea, a migratory stopover location. Our results show species-level differences in migration timing and behavior for these common and widespread species, demonstrating the need to consider their unique temporal and spatial movement ecology when incorporating wild birds into AIV risk modeling and management.

Frontiers in Ecology and Evolution↗
USGS2021

Comparative genomics analyses support the reclassification of Bisgaard taxon 40 as Mergibacter gen. nov., with Mergibacter septicus sp. nov. as type species: Novel insights into the phylogeny and virulence factors of a Pasteurellaceae family member associated with mortality events in seabirds

The Pasteurellaceae family has been associated with fatal diseases in numerous avian species. Several new taxa within this family, including Bisgaard taxon 40, have been recently described in wild birds, but their genomic characteristics and pathogenicity are not well understood. We isolated Bisgaard taxon 40 from four species of seabirds, including one sampled during a mass, multi-species mortality event in Florida, United States. Here, we present a comprehensive phenotypic and genetic characterization of Bisgaard taxon 40 and comparative genomic analysis with reference strains from the Pasteurellaceae family, aiming at determining its phylogenetic position, antimicrobial susceptibility profile, and identifying putative virulence factors. In silico multilocus sequence-based and whole-genome-based phylogenetic analysis clustered all Bisgaard taxon 40 strains together on a distinct branch separated from the other members of the Pasteurellaceae family, indicating that Bisgaard taxon 40 could represent a new genus. These findings were further supported by protein similarity analyses using the concatenation of 31 conserved proteins and other taxonomic approaches such as the percentage of conserved protein test. Additionally, several putative virulence factors were identified, including those associated with adhesion (capsule, ompA , ompH ) and colonization ( exbD , fur , galU , galE , lpxA , lpxC , and kdsA ) of the host and a cytolethal distending toxin ( cdt ), which may have played a role in disease development leading to the mortality event. Considerably low minimum inhibitory concentrations (MICs) were found for all the drugs tested, in concordance with the absence of antimicrobial resistance genes in these genomes. The novel findings of this study highlight genomic and phenotypic characteristics of this bacterium, providing insights into genome evolution and pathogenicity. We propose a reclassification of these organisms within the Pasteurellaceae family, designated as Mergibacter gen. nov., with Mergibacter septicus sp. nov. as the type species. The type strain is Mergibacter septicus A25201 T (=DSM 112696).

Frontiers in Microbiology↗
USGS2025

Bacterial community diversity and potential eco-physiological roles in toxigenic blooms composed of Microcystis, Aphanizomenon or Planktothrix

Cyanobacterial toxicity, cyanotoxins, and their impact on aquatic ecosystems and human health are well documented. In comparison, less is known about bloom-associated bacterial communities. Co-occurring bacteria can influence bloom development, physiology and collapse, and may also provide a niche for pathogenic bacteria. Existing research focuses on the cyanosphere of Microcystis -dominated blooms, despite the increasing prevalence of filamentous genera ( Aphanizomenon and Planktothrix ). This pilot study aimed to broaden our understanding of the bacterial consortia attached to morphologically distinct cyanobacteria (coccoid and filamentous) dominating phytoplankton communities and to explore their potential roles in amplifying the impacts of cyanobacterial blooms. We investigated four shallow freshwater bodies across three continents and two climate zones: an urban pond in the USA, a dammed reservoir and a natural lake in Poland, and an urban water body in Singapore. Amplicon sequencing (16S rRNA gene) was used to characterize bacterial communities, while shotgun metagenomics identified nitrogen- and phosphorus-cycling genes to infer potential eco-physiological functions. Cyanobacteria dominated bacterioplankton assemblages at all sites (>35.6%), with bloom composition influencing toxigenic profiles. A mixed bloom of Microcystis , Snowella , and Aphanizomenon had the broadest range of cyanotoxin synthetase genes ( mcy E, cyr J, ana F and sxt A). Microcystis blooms correlated with increased Roseomonas , while Planktothrix co-occurred with Flavobacterium – both bacteria likely contribute to nutrient-cycling within blooms and represent potential opportunistic pathogens for aquatic organisms and humans. The Microcystis cyanosphere exhibited the highest number of significant positive correlations with bacteria (19 relations), compared to Planktothrix and Aphanizomenon (11 and 2 relations, respectively). Non-diazotrophic blooms of Microcystis and Planktothrix showed greater abundances of nitrogen – ( ure B, gln A, nar B, and nar HZ) and phosphorus-cycling genes ( pho BHPR and ppk 1), indicating a strong dependence on associated bacteria for nutrient acquisition compared to diazotrophic Aphanizomenon . These findings suggest that Aphanizomenon -dominated blooms may be sustained by simpler microbiomes. Our results provide preliminary evidence of cyanosphere heterogeneity potentially shaped by the dominance or coexistence of three morphologically and eco-physiologically distinct genera of cyanobacteria. A comprehensive knowledge of the taxonomy and functional roles of bloom-associated microbiomes is therefore essential to understand bloom activity, evaluate the environmental threat, and develop effective strategies for prevention and mitigation.

Frontiers in Microbiology↗
USGS2022

Susceptibility of beavers to chronic wasting disease

Chronic wasting disease (CWD) is a contagious, fatal, neurodegenerative prion disease of cervids. The expanding geographical range and rising prevalence of CWD are increasing the risk of pathogen transfer and spillover of CWD to non-cervid sympatric species. As beavers have close contact with environmental and food sources of CWD infectivity, we hypothesized that they may be susceptible to CWD prions. We evaluated the susceptibility of beavers to prion diseases by challenging transgenic mice expressing beaver prion protein (tgBeaver) with five strains of CWD, four isolates of rodent-adapted prions and one strain of Creutzfeldt–Jakob disease. All CWD strains transmitted to the tgBeaver mice, with attack rates highest from moose CWD and the 116AG and H95+ strains of deer CWD. Mouse-, rat-, and especially hamster-adapted prions were also transmitted with complete attack rates and short incubation periods. We conclude that the beaver prion protein is an excellent substrate for sustaining prion replication and that beavers are at risk for CWD pathogen transfer and spillover.

Biology↗
USGS2022

Plague and trace metals in natural systems

All pathogenic organisms are exposed to abiotic influences such as the microclimates and chemical constituents of their environments. Even those pathogens that exist primarily within their hosts or vectors can be influenced directly or indirectly. Yersinia pestis , the flea-borne bacterium causing plague, is influenced by climate and its survival in soil suggests a potentially strong influence of soil chemistry. We summarize a series of controlled studies conducted over four decades in Russia by Dr. Evgeny Rotshild and his colleagues that investigated correlations between trace metals in soils, plants, and insects, and the detection of plague in free-ranging small mammals. Trace metal concentrations in plots where plague was detected were up to 20-fold higher or lower compared to associated control plots, and these differences were >2-fold in 22 of 38 comparisons. The results were statistically supported in eight studies involving seven host species in three families and two orders of small mammals. Plague tended to be positively associated with manganese and cobalt, and the plague association was negative for copper, zinc, and molybdenum. In additional studies, these investigators detected similar connections between pasturellosis and concentrations of some chemical elements. A One Health narrative should recognize that the chemistry of soil and water may facilitate or impede epidemics in humans and epizootics in non-human animals.

International Journal of Environmental Research an↗
USGS2023

Informing ASR treatment practices in a Florida aquifer through a human health risk approach

Aquifer storage and recovery (ASR) can augment water supplies and hydrologic flows under varying climatic conditions. However, imposing drinking water regulations on ASR practices, including pre-treatment before injection into the aquifer, remains arguable. Microbial inactivation data— Escherichia coli , Pseudomonas aeruginosa , poliovirus type 1 and Cryptosporidium parvum —were used in a human health risk assessment to identify how the storage time of recharged water in the Floridan Aquifer enhances pathogen inactivation, thereby mitigating the human health risks associated with ingestion. We used a quantitative microbial risk assessment to evaluate the risks for a gastrointestinal infection (GI) and the associated disability-adjusted life years (DALYs) per person per year. The risk of developing a GI infection for drinking water no longer exceeded the suggested annual risk threshold (1 × 10 −4 ) by days 31, 1, 52 and 80 for each pathogen, respectively. DALYs per person per year no longer exceeded the World Health Organization threshold (1 × 10 −6 ) by days 27, <1, 43 and 72. In summary, storage time in the aquifer yields a significant reduction in health risk. The findings emphasize that considering microbial inactivation, caused by storage time and geochemical conditions within ASR storage zones, is critical for recharge water treatment processes.

Florida↗
USGS2021

Soil reservoir dynamics of ophidiomyces ophidiicola, the causative agent of snake fungal disease

Wildlife diseases pose an ever-growing threat to global biodiversity. Understanding how wildlife pathogens are distributed in the environment and the ability of pathogens to form environmental reservoirs is critical to understanding and predicting disease dynamics within host populations. Snake fungal disease (SFD) is an emerging conservation threat to North American snake populations. The causative agent, Ophidiomyces ophidiicola (Oo), is detectable in environmentally derived soils. However, little is known about the distribution of Oo in the environment and the persistence and growth of Oo in soils. Here, we use quantitative PCR to detect Oo in soil samples collected from five snake dens. We compare the detection rates between soils collected from within underground snake hibernacula and associated, adjacent topsoil samples. Additionally, we used microcosm growth assays to assess the growth of Oo in soils and investigate whether the detection and growth of Oo are related to abiotic parameters and microbial communities of soil samples. We found that Oo is significantly more likely to be detected in hibernaculum soils compared to topsoils. We also found that Oo was capable of growth in sterile soil, but no growth occurred in soils with an active microbial community. A number of fungal genera were more abundant in soils that did not permit growth of Oo, versus those that did. Our results suggest that soils may display a high degree of both general and specific suppression of Oo in the environment. Harnessing environmental suppression presents opportunities to mitigate the impacts of SFD in wild snake populations.

Journal of Fungi↗
USGS2018

Fungal disease prevention in seedlings of rice (Oryza sativa) and other grasses by growth-promoting seed-associated endophytic bacteria from invasive Phragmites australis

Non-cultivated plants carry microbial endophytes that may be used to enhance development and disease resistance of crop species where growth-promoting and protective microbes may have been lost. During seedling establishment, seedlings may be infected by several fungal pathogens that are seed or soil borne. Several species of Fusarium , Pythium and other water moulds cause seed rots during germination. Fusarium blights of seedlings are also very common and significantly affect seedling development. In the present study we screened nine endophytic bacteria isolated from the seeds of invasive Phragmites australis by inoculating onto rice, Bermuda grass ( Cynodon dactylon ), or annual bluegrass ( Poa annua ) seeds to evaluate plant growth promotion and protection from disease caused by Fusarium oxysporum . We found that three bacteria belonging to genus Pseudomonas spp. (SLB4- P. fluorescens , SLB6- Pseudomonas sp. and SY1- Pseudomonas sp.) promoted seedling development, including enhancement of root and shoot growth, and stimulation of root hair formation. These bacteria were also found to increase phosphate solubilization in in vitro experiments. Pseudomonas sp. (SY1) significantly protected grass seedlings from Fusarium infection. In co-culture experiments, strain SY1 strongly inhibited fungal pathogens with 85.71% growth inhibition of F. oxysporum , 86.33% growth inhibition of Curvularia sp. and 82.14% growth inhibition of Alternaria sp. Seedlings previously treated with bacteria were found much less infected by F. oxysporum in comparison to non-treated controls. On microscopic observation we found that bacteria appeared to degrade fungal mycelia actively. Metabolite products of strain SY1 in agar were also found to inhibit fungal growth on nutrient media. Pseudomonas sp. (SY1) was found to produce antifungal volatiles. Polymerase chain reaction (PCR) amplification using specific primers for pyrrolnitirin synthesis and HCN (hydrogen cyanide) production suggested presence of genes for both compounds in the genome of SY1. HCN was detected in cultures of SY1. We conclude that microbes from non-cultivated plants may provide disease protection and promote growth of crop plants.

Microorganisms↗
USGS2020

Growth and behavior of North American microbes on Phragmites australis leaves

Phragmites australis subsp. australis is a cosmopolitan wetland grass that is invasive in many regions of the world, including North America, where it co-occurs with the closely related Phragmites australis subsp. americanus. Because the difference in invasive behavior is unlikely to be related to physiological differences, we hypothesize that interactions with unique members of their microbiomes may significantly affect the behavior of each subspecies. Therefore, we systematically inoculated both plant lineages with a diverse array of 162 fungal and bacterial isolates to determine which could (1) differentiate between Phragmites hosts, (2) infect leaves at various stages of development, or (3) obtain plant-based carbon saprophytically. We found that many of the microbes isolated from Phragmites leaves behave as saprophytes. Only 1% (two taxa) were determined to be strong pathogens, 12% (20 taxa) were weakly pathogenic, and the remaining 87% were nonpathogenic. None of the isolates clearly discriminated between host plant lineages, and the Phragmites cuticle was shown to be a strong nonspecific barrier to infection. These results largely agree with the broad body of literature on leaf-associated phyllosphere microbes in Phragmites.

Microorganisms↗
USGS2023

Monkeypox virus in animals: Current knowledge of viral transmission and pathogenesis in wild animal reservoirs and captive animal models

Mpox, formerly called monkeypox, is now the most serious orthopoxvirus (OPXV) infection in humans. This zoonotic disease has been gradually re-emerging in humans with an increasing frequency of cases found in endemic areas, as well as an escalating frequency and size of epidemics outside of endemic areas in Africa. Currently, the largest known mpox epidemic is spreading throughout the world, with over 85,650 cases to date, mostly in Europe and North America. These increased endemic cases and epidemics are likely driven primarily by decreasing global immunity to OPXVs, along with other possible causes. The current unprecedented global outbreak of mpox has demonstrated higher numbers of human cases and greater human-to-human transmission than previously documented, necessitating an urgent need to better understand this disease in humans and animals. Monkeypox virus (MPXV) infections in animals, both naturally occurring and experimental, have provided critical information about the routes of transmission; the viral pathogenicity factors; the methods of control, such as vaccination and antivirals; the disease ecology in reservoir host species; and the conservation impacts on wildlife species. This review briefly described the epidemiology and transmission of MPXV between animals and humans and summarizes past studies on the ecology of MPXV in wild animals and experimental studies in captive animal models, with a focus on how animal infections have informed knowledge concerning various aspects of this pathogen. Knowledge gaps were highlighted in areas where future research, both in captive and free-ranging animals, could inform efforts to understand and control this disease in both humans and animals.

Viruses↗
USGS2011

Rapid diagnosis of avian influenza virus in wild birds: Use of a portable rRT-PCR and freeze-dried reagents in the field

Wild birds have been implicated in the spread of highly pathogenic avian influenza (HPAI) of the H5N1 subtype, prompting surveillance along migratory flyways. Sampling of wild birds for avian influenza virus (AIV) is often conducted in remote regions, but results are often delayed because of the need to transport samples to a laboratory equipped for molecular testing. Real-time reverse transcriptase polymerase chain reaction (rRT-PCR) is a molecular technique that offers one of the most accurate and sensitive methods for diagnosis of AIV. The previously strict lab protocols needed for rRT-PCR are now being adapted for the field. Development of freeze-dried (lyophilized) reagents that do not require cold chain, with sensitivity at the level of wet reagents has brought on-site remote testing to a practical goal. Here we present a method for the rapid diagnosis of AIV in wild birds using an rRT-PCR unit (Ruggedized Advanced Pathogen Identification Device or RAPID, Idaho Technologies, Salt Lake City, UT) that employs lyophilized reagents (Influenza A Target 1 Taqman; ASAY-ASY-0109, Idaho Technologies). The reagents contain all of the necessary components for testing at appropriate concentrations in a single tube: primers, probes, enzymes, buffers and internal positive controls, eliminating errors associated with improper storage or handling of wet reagents. The portable unit performs a screen for Influenza A by targeting the matrix gene and yields results in 2-3 hours. Genetic subtyping is also possible with H5 and H7 primer sets that target the hemagglutinin gene. The system is suitable for use on cloacal and oropharyngeal samples collected from wild birds, as demonstrated here on the migratory shorebird species, the western sandpiper (Calidrus mauri) captured in Northern California. Animal handling followed protocols approved by the Animal Care and Use Committee of the U.S. Geological Survey Western Ecological Research Center and permits of the U.S. Geological Survey Bird Banding Laboratory. The primary advantage of this technique is to expedite diagnosis of wild birds, increasing the chances of containing an outbreak in a remote location. On-site diagnosis would also prove useful for identifying and studying infected individuals in wild populations. The opportunity to collect information on host biology (immunological and physiological response to infection) and spatial ecology (migratory performance of infected birds) will provide insights into the extent to which wild birds can act as vectors for AIV over long distances.

Journal of Visualized Experiments↗