Detection and isolation of H5N1 influenza virus from large volumes of natural water

Khalenkov, Alexey M.; Laver, W.G.; Webster, Robert G.

doi:10.1016/j.jviromet.2008.01.001

Cited by 51 publications

(50 citation statements)

References 15 publications

(15 reference statements)

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“…A simple technique for detection and isolation of H5N1 AIV from large volumes of natural water was described recently (15). In that study, particles of a nonpathogenic reverse-genetic virus carrying the HA and neuraminidase (NA) genes of an H5N1 virus were isolated from water using fixed CRBCs and ECEs.…”

Section: Discussionmentioning

confidence: 99%

“…The 4-guanidino-Neu5Ac2en stock solutions were prepared using Relenza (GlaxoSmithKline, Evreux, France) Rotadisks (5 mg of zanamivir with lactose) by dissolving 5-mg blister capsules in 285 l of phosphate-buffered saline (PBS) and were stored at Ϫ20°C. The CRBCs were formalin fixed as previously described (15). After fixation, the erythrocyte solution was washed eight times, and it could be stored for up to 6 months at 4°C in PBS as a 10% (vol/vol) suspension containing antibiotics and antimycotics at the following final concentrations: 200 U/ml penicillin G, 200 g/ml streptomycin sulfate, and 0.5 g/ml amphotericin.…”

Section: Methodsmentioning

confidence: 99%

“…The most sensitive detection methods are virus isolation in embryonated specific-pathogen-free (SPF) chicken eggs and reverse transcription (RT)-PCR (15). Isolation in SPF eggs is highly sensitive for detection of infectious virus particles.…”

Section: Routes Of Avian Influenza Virus (Aiv) Dispersal Among Aquatimentioning

confidence: 99%

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Detection and Quantification of Infectious Avian Influenza A (H5N1) Virus in Environmental Water by Using Real-Time Reverse Transcription-PCR

Dovas

Papanastassopoulou

Georgiadis

et al. 2010

Appl Environ Microbiol

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Routes of avian influenza virus (AIV) dispersal among aquatic birds involve direct (bird-to-bird) and indirect (waterborne) transmission. The environmental persistence of H5N1 virus in natural water reservoirs can be assessed by isolation of virus in embryonated chicken eggs. Here we describe development and evaluation of a real-time quantitative reverse transcription (RT)-PCR (qRT-PCR) method for detection of H5N1 AIV in environmental water. This method is based on adsorption of virus particles to formalin-fixed erythrocytes, followed by qRT-PCR detection. The numbers of hemagglutinin RNA copies from H5N1 highly pathogenic AIV particles adsorbed to erythrocytes detected correlated highly with the infectious doses of the virus that were determined for three different types of artificially inoculated environmental water over a 17-day incubation period. The advantages of this method include detection and quantification of infectious H5N1 AIVs with a high level of sensitivity, a wide dynamic range, and reproducibility, as well as increased biosecurity. The lowest concentration of H5N1 virus that could be reproducibly detected was 0.91 50% egg infective dose per ml. In addition, a virus with high virion stability (Tobacco mosaic virus) was used as an internal control to accurately monitor the efficiency of RNA purification, cDNA synthesis, and PCR amplification for each individual sample. This detection system could be useful for rapid high-throughput monitoring for the presence of H5N1 AIVs in environmental water and in studies designed to explore the viability and epidemiology of these viruses in different waterfowl ecosystems. The proposed method may also be adapted for detection of other AIVs and for assessment of their prevalence and distribution in environmental reservoirs.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Detection and Quantification of Infectious Avian Influenza A (H5N1) Virus in Environmental Water by Using Real-Time Reverse Transcription-PCR

Dovas

Papanastassopoulou

Georgiadis

et al. 2010

Appl Environ Microbiol

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“…Compared to detection of AIV via molecular methods, isolation of viable virus from natural lake water has occurred even less frequently (4,11). Low rates of AIV detection and isolation from natural surface water, combined with recent advances to concentrate large volumes of water for detection of AIV (14)(15)(16), demonstrate that even amid favorable conditions (i.e., high concentrations of aquatic birds and active viral shedding), AIV likely exists at low concentrations in water. As such, any factor that acts to increase virus concentrations and facilitate ingestion by birds could play an important role in transmission.…”

mentioning

confidence: 99%

Accumulation and Inactivation of Avian Influenza Virus by the Filter-Feeding Invertebrate Daphnia magna

Meixell

Borchardt

Spencer

2013

Appl Environ Microbiol

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The principal mode of avian influenza A virus (AIV) transmission among wild birds is thought to occur via an indirect fecal-oral route, whereby individuals are exposed to virus from the environment through contact with virus-contaminated water. AIV can remain viable for an extended time in water; however, little is known regarding the influence of the biotic community (i.e., aquatic invertebrates) on virus persistence and infectivity in aquatic environments. We conducted laboratory experiments to investigate the ability of an aquatic filter-feeding invertebrate, Daphnia magna, to accumulate virus from AIV-dosed water under the hypothesis that they represent a potential vector of AIV to waterfowl hosts. We placed live daphnids in test tubes dosed with low-pathogenicity AIV (H3N8 subtype isolated from a wild duck) and sampled Daphnia tissue and the surrounding water using reverse transcription-quantitative PCR (RT-qPCR) at 3-to 120-min intervals for up to 960 min following dosing. Concentrations of viral RNA averaged 3 times higher in Daphnia tissue than the surrounding water shortly after viral exposure, but concentrations decreased exponentially through time for both. Extracts from Daphnia tissue were negative for AIV by cell culture, whereas AIV remained viable in water without Daphnia present. Our results suggest daphnids can accumulate AIV RNA and effectively remove virus particles from water. Although concentrations of viral RNA were consistently higher in Daphnia tissue than the water, additional research is needed on the time scale of AIV inactivation after Daphnia ingestion to fully elucidate Daphnia's role as a potential vector of AIV infection to aquatic birds.

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“…Both direct and concentrated samples have been used with either isolation or molecular detection. It is more likely that influenza virus will be detected when environmental water samples are concentrated [63,64]. Feces from infected birds contain high levels of virus [9] and do not require concentration.…”

Section: Discussionmentioning

confidence: 99%

Detection of Influenza A viruses at migratory bird stopover sites in Michigan, USA

Lickfett

Clark

Gehring

et al. 2018

Infection Ecology & Epidemiology

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Introduction: Influenza A viruses have the potential to cause devastating illness in humans and domestic poultry. Wild birds are the natural reservoirs of Influenza A viruses and migratory birds are implicated in their global dissemination. High concentrations of this virus are excreted in the faeces of infected birds and faecal contamination of shared aquatic habitats can lead to indirect transmission among birds via the faecal-oral route. The role of migratory birds in the spread of avian influenza has led to large-scale surveillance efforts of circulating avian influenza viruses through direct sampling of live and dead wild birds. Environmental monitoring of bird habitats using molecular detection methods may provide additional information on the persistence of influenza virus at migratory stopover sites distributed across large spatial scales. Materials and methods: In the current study, faecal and water samples were collected at migratory stopover sites and evaluated for Influenza A by real-time quantitative reverse transcriptase PCR. Results and Discussion: This study found that Influenza A was detected at 53% of the evaluated stopover sites, and 7% and 4.8% of the faecal and water samples, respectively, tested positive for Influenza A virus. Conclusion: Environmental monitoring detected Influenza A at stopover sites used by migratory birds.

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Detection and isolation of H5N1 influenza virus from large volumes of natural water

Cited by 51 publications

References 15 publications

Detection and Quantification of Infectious Avian Influenza A (H5N1) Virus in Environmental Water by Using Real-Time Reverse Transcription-PCR

Detection and Quantification of Infectious Avian Influenza A (H5N1) Virus in Environmental Water by Using Real-Time Reverse Transcription-PCR

Accumulation and Inactivation of Avian Influenza Virus by the Filter-Feeding Invertebrate Daphnia magna

Detection of Influenza A viruses at migratory bird stopover sites in Michigan, USA

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