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Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

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  • Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

    The highly pathogenic H5N1 is a major avian pathogen that crosses species barriers and seriously affects humans as well as some mammals. It mutates in an intensified manner and is considered a potential candidate for the possible next pandemic with all the catastrophic consequences.

    Methods: Nasal swabs were collected from donkeys suffered from respiratory distress.

    The virus was isolated from the pooled nasal swabs in specific pathogen free embryonated chicken eggs (SPF-ECE). Reverse transcriptase polymerase chain reaction (RT-PCR) and sequencing of both haemagglutingin and neuraminidase were performed.H5 seroconversion was screened using haemagglutination inhibition (HI) assay on 105 donkey serum samples.
    Results: We demonstrated that H5N1 jumped from poultry to another mammalian host; donkeys. Phylogenetic analysis showed that the virus clustered within the lineage of H5N1 from Egypt, closely related to 2009 isolates.
    It harboured few genetic changes compared to the closely related viruses from avian and humans. The neuraminidase lacks oseltamivir resistant mutations.Interestingly, HI screening for antibodies to H5 haemagglutinins in donkeys revealed high exposure rate.

    Conclusions: These findings extend the host range of the H5N1 influenza virus, possess implications for influenza virus epidemiology and highlight the need for the systematic surveillance of H5N1 in animals in the vicinity of backyard poultry units especially in endemic areas.

    Author: Ahmed Abdel-MoneimAhmad Abdel-GhanySalama Shany
    Credits/Source: Journal of Biomedical Science 2010, 17:25http://7thspace.com/headlines/341392...m_donkeys.html
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  • #2
    Re: Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

    The report mentioned is now available and opens on the link below.

    AVIAN INFLUENZA (24): EGYPT, DONKEY***********************************A ProMED-mail post<http://www.promedmail.org>ProMED-mail is a program of theInternational Society for Infectious Diseases<http://www.isid.org> Date: Wed 14 Apr 2010Source: Journal of Biomedical Science 2010, 17:25 [edited] Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys - Provisional abstract----------------------------Highly pathogenic H5N1 is a major avian pathogen that crosses species barriers and seriously affects humans as well as some mammals. It mutates in an intensified manner and is considered a potential candidate for the possible next pandemic with all the catastrophic consequences. Methods------------Nasal swabs were collected from donkeys suffering from respiratory distress. The virus was isolated from the pooled nasal swabs in specific pathogen free embryonated chicken eggs (SPF-ECE). Reverse transcriptase polymerase chain reaction (RT-PCR) and sequencing of both haemagglutingin and neuraminidase were performed. H5 seroconversion was screened using haemagglutination inhibition (HI) assay on 105 donkey serum samples. Results----------We demonstrated that H5N1 jumped from poultry to another mammalian host, donkeys. Phylogenetic analysis showed that the virus clustered within the lineage of H5N1 from Egypt closely related to 2009 isolates. It harboured few genetic changes compared to the closely related viruses from avian and humans. The neuraminidase lacks oseltamivir resistant mutations. Interestingly, HI screening for antibodies to H5 haemagglutinins in donkeys revealed a high exposure rate. Conclusions------------These findings extend the host range of the H5N1 influenza virus, possess implications for influenza virus epidemiology, and highlight the need for the systematic surveillance of H5N1 in animals in the vicinity of backyard poultry units, especially in endemic areas. [Byline: Ahmed S Abdel-Moneim, Ahmad E Abdel-Ghany, Salama AS Shany] --Communicated by:ProMED-mail <promed@promedmail.org> [The donkey has already been added to the other 16 animals of the class Mammalia included in the USGS List of Species affected by H5N1 (Avian Influenza), (referenced reports of highly pathogenic avian influenza H5N1 in wildlife and domestic animals), which is available at <http://www.nwhc.usgs.gov/disease_information/avian_influenza/affected_species_chart.jsp>. For the full paper from which the abstract has been copied, see: Ref 1. Ahmed S Abdel-Moneim, Ahmad E Abdel-Ghany & Salama AS Shany (2010). Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys. Journal of Biomedical Science 2010, 17:25; doi:10.1186/1423-0127-17-25;<http://www.jbiomedsci.com/content/pdf/1423-0127-17-25.pdf>. - Mod.AS]


    24th March 2009, 1wk after an outbreak of H5N1 infection in poultry in the village, where many donkeys suffered from the same clinical manifestations in an epidemic manner. The virus was isolated from a pool of nasal discharge from three affected animals. It produced haemagglutination only
    after the 3rd egg passage.

    RT-PCR was performed to the full length of both NA and
    HA genes, where they were sequenced directly after gel purification. Sequences were deposited in GenBank under accession numbers; GU371911 and GU371912 for HA and NA respectively. The HA and NA genes of the investigated equine isolates revealed that they belonged to (5J), (1J) lineages respectively (According to the Influenza A Virus Genotype Tool) [24]. Phylogenetic analyses revealed that the HA of the equine isolate related to sublineages, A (A1) (Fig.1). The equine isolate showed a typical polybasic cleavage motif with the GERRRKKR*GLF consensus sequence found
    in clade 2.2 viruses. It also contains amino acid D403 characteristic to sub-clade 2.2.1

    (Fig.2). The haemagglutinin gene was found to be closely related to
    A/chicken/Egypt/0894-NLQP/2008, A/Egypt/N00605/2009 and
    A/chicken/Egypt/092-NLQP/2009 while the neuraminidase gene of the current
    strain is closely related to A/Egypt/N03450/2009 and A/Egypt/N05056/2009.
    However, none of these strains were isolated from localities near to Beni-Suef.

    The equine isolate has amino acids Q226 and G228 (H3 influenza numbering)
    denoting the preferential binding of α-2,3 linkage, typical for the avian and equine viruses but not human ones [5, 25]. This finding demonstrated that the isolates with avian specific receptor binding properties can replicate and cause infection in equines. Different amino-acids that are implicated in receptor specificity Y98, S136,W153, H183, E190, K193 L194 E216 P221 K222 G225, Q226, S227,G228 (H3 influenza

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    • #3
      Re: Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

      Another interesting comment from the .pdf:

      Although the disease did not constitute a real threat to donkeys, it raises the concern of different issues including the route of transmission to donkeys, whether being from aerosol exposure of pulverized infected birds droppings or contaminated feeds and water or because of contact with infected birds.
      The salvage of human life ought to be placed above barter and exchange ~ Louis Harris, 1918

      Comment


      • #4
        Re: Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

        I remember the tigers, cats, stone marten etc.

        they got it from eating contaminated meat

        I don't remember having heard about dust from droppings to transmit
        H5N1
        Although - how does it transmit on these poultry farms, that
        _must_ be known ?!?!?! I assume respiratory, breathing chickens

        within a few day all chickens dead
        I'm interested in expert panflu damage estimates
        my current links: http://bit.ly/hFI7H ILI-charts: http://bit.ly/CcRgT

        Comment


        • #5
          Re: Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

          So we know when they said the pigs did not have this virus they did!
          we also know now it has spread into the donkeys within Egypt at least one time.even thou spreading outside of poultry was denied in the past!

          we also know quite a few horses and donkeys were located dead in Giza during mid 2009 and the cause was never made public as i recall.

          I'm wonder if this report is what generated the concern here last year for h3n8

          a pretty massive campaign is currently under way to cull stray dogs and cats through out Egypt. due to rabies they claim, which i have not heard much about at all, or heard only a little about via the normal media.

          I'm sure it can be spread by winds and carried on dust and we have dust storms quite often. or even picked up in droppings from other animals as they graze or forage.the animals are often even housed in the same areas.

          Comment


          • #6
            Re: Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

            from 2008 - Reported as H3N8

            "The Commission began an emergency in the central laboratories of the Ministry of Agriculture, in collaboration with the Directorate of Veterinary Medicine in Aswan work yesterday, which is surrounded flu horses after the emergence of 165 cases of infection among horses and donkeys at the centre of Edfu"


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            • #7
              Re: Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys


              Submitted (10-SEP-2008) Viral and Zoonotic Diseases Research
              Program, U.S. Naval Medical Research Unit No. 3, Extension of
              Ramses Street, Cairo 11517, Egypt
              /serotype="H3N8"
              /isolate="Alex-1"
              /host="Equus caballus"
              /collection_date="Jun-2008"


              Epizootiological and molecular characterization of equine influenza
              virus 2008 outbreak in Egypt
              Submitted (06-OCT-2009)
              serotype="H3N8"
              /host="equine"
              /collection_date="Jul-2008"


              Isolation and characterization of highly pathogenic avian influenza
              virus subtype H5N1 from donkeys
              Submitted (01-JAN-2010)
              /serotype="H5N1"
              /host="equine"
              /country="Egypt: Beni-Suef Governorate, Aborady Village"
              /collection_date="24-Mar-2009"
              I'm interested in expert panflu damage estimates
              my current links: http://bit.ly/hFI7H ILI-charts: http://bit.ly/CcRgT

              Comment


              • #8
                Re: Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys

                in addition the equine isolate lacks aa S145. This deletion is also present in all other
                viruses grouped into 2.2 sublineage A1 which also includes sequences from human
                H5N1 isolates (Fig.2). The significance of this deletion is unknown, but it should be
                noted that this position is close to a domain modulating receptor interaction.
                Interestingly, strains with this deletion appear to evolve towards a receptor usage
                that is similar to that of the seasonal H1N1[31,Veljkovic et.al. 2009]

                ...

                (4-6 months after virus isolation)
                27 out of 105 of the examined animals were H5 positive with the
                highest percentage found in the area where the virus was isolated


                I'm interested in expert panflu damage estimates
                my current links: http://bit.ly/hFI7H ILI-charts: http://bit.ly/CcRgT

                Comment

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