Ahead of Print -Active Surveillance for Avian Influenza Virus, Egypt, 2010–2012 - Volume 20, Number 4—April 2014 - Emerging Infectious Disease journal - CDC

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Ahead of Print -Active Surveillance for Avian Influenza Virus, Egypt, 2010–2012 - Volume 20, Number 4—April 2014 - Emerging Infectious Disease journal - CDC

Volume 20, Number 4—April 2014

Research

Active Surveillance for Avian Influenza Virus, Egypt, 2010–2012

Article Contents

• Materials and Methods
• Results
• Discussion
• Acknowledgment
• References
• Figure 1
• Figure 2
• Figure 3
• Figure 4
• Figure 5
• Table 1
• Table 2
• Table 3
• Technical Appendix - 461 KB 
• Suggested Citation

Ghazi Kayali , Ahmed Kandeil, Rabeh El-Shesheny, Ahmed S. Kayed, Mokhtar M. Gomaa, Asmaa M. Maatouq, Mahmoud M. Shehata, Yassmin Moatasim, Ola Bagato, Zhipeng Cai, Adam Rubrum, Mohamed A. Kutkat, Pamela P. McKenzie, Robert G. Webster, Richard J. Webby, and Mohamed A. Ali

Author affiliations: St. Jude Children's Research Hospital, Memphis, Tennessee, USA (G. Kayali, A. Rubrum, P.P. McKenzie, R.G. Webster, R.J. Webby); National Research Center, Giza, Egypt (A. Kandeil, R. El-Shesheny, A.S. Kayed, M.M. Gomaa, A.M. Maatouq, M.M. Shehata, Y. Moatasim, O. Bagato, M.A. Kutkat, M.A. Ali);Georgia State University, Atlanta, Georgia, USA (Z. Cai)

Suggested citation for this article

Abstract

Continuous circulation of influenza A(H5N1) virus among poultry in Egypt has created an epicenter in which the viruses evolve into newer subclades and continue to cause disease in humans. To detect influenza viruses in Egypt, since 2009 we have actively surveyed various regions and poultry production sectors. From August 2010 through January 2013, >11,000 swab samples were collected; 10% were positive by matrix gene reverse transcription PCR. During this period, subtype H9N2 viruses emerged, cocirculated with subtype H5N1 viruses, and frequently co-infected the same avian host. Genetic and antigenic analyses of viruses revealed that influenza A(H5N1) clade 2.2.1 viruses are dominant and that all subtype H9N2 viruses are G1-like. Cocirculation of different subtypes poses concern for potential reassortment. Avian influenza continues to threaten public and animal health in Egypt, and continuous surveillance for avian influenza virus is needed.

In 2008, highly pathogenic avian influenza (HPAI) A(H5N1) virus became enzootic among poultry in Egypt, and the country became an epicenter for virus activity (1). As the established viruses drifted over time, viral genetic and antigenic diversity was generated. During 2010–2011, subclade 2.2.1 viruses (direct-drift progeny of the initially introduced virus) and 2.2.1.1 viruses (which might have emerged because of vaccine pressure) were cocirculating among poultry in Egypt (2). These subclades differed genetically and antigenically, hence complicating control efforts, especially vaccination (3). Subclade 2.2.1 viruses, commonly isolated from backyard flocks that are not vaccinated, caused all of the human cases in Egypt; from 2006 through September 2013, the toll rose to 173 cases and 63 deaths (4,5). Subclade 2.2.1.1 viruses were more prevalent on commercial farms, where vaccines are more frequently used (6). Furthermore, recent reports have indicated that very few mutations are needed for subtype H5N1 to become transmissible among ferrets, the best mammalian model of human influenza infection (7,8). In Egypt, a subtype H5N1 virus was found to have 2 of the 4 mutations needed to gain the transmissibility function, thereby underlying the need and urgency for surveillance among poultry (8). The Nile Delta region of Egypt was also identified as an area where substantial reassortment of influenza viruses can take place (9). As a further complication, in 2011, subtype H9N2 viruses were detected in poultry from areas in Egypt where subtype H5N1 viruses circulate (10).

Since 2009, we have been conducting systematic, active surveillance of avian influenza virus (AIV) among poultry in Egypt; the same locations are sampled over time, regardless of whether a clinical outbreak of disease is present. We previously reported that the threat of HPAI (H5N1) virus is widespread beyond rural areas and that the commercial sector is a key reservoir for virus transmission (11). Here we provide an update on the changing epizootiology and genetic features of AIV in Egypt and report co-infection of poultry in Egypt with influenza virus subtypes H5N1 and H9N2.

Materials and Methods

Sample Collection and Processing

Figure 1

Figure 1. . Location of surveillance governorates and percentage of avian influenza virus detection in each governorate, Egypt, 2010–2012.

A team of veterinarians collected cloacal and oropharyngeal swab samples from 11,452 birds from 4 poultry production sectors: commercial farms, backyard flocks, live-bird markets, and abattoirs. One swab sample was collected per bird, and depending on the size of the population, as many as 5 birds were sampled per flock. Birds were not randomly selected; samples were also collected from sick or dead birds found on site. From August 2010 through January 2013, a total of 6,904 cloacal and 4,548 oropharyngeal samples were collected from 63 sites in 7 governorates in Egypt, including Cairo (4 neighborhoods); 4 Nile Delta governorates (Qalubiya [12 villages], Menofiya [9 villages], Sharqiya [3 towns], and Daqahliya [4 towns]); and 2 mid-Egypt governorates (Fayyoum [22 villages] and BeniSuef [9 villages]) (Figure 1). The selected governorates represent the main foci of the poultry industry in Egypt and sites of previous AIV detection (11). The selected sampling sites were areas at which the veterinarian was known to the local population and thus had access to the poultry. The sites were routinely visited on a monthly basis regardless of the occurrence of clinical signs or poultry deaths. Study veterinarians subjectively recorded their field observations. Swab samples were collected in medium containing 50% glycerol, 50% phosphate-buffered saline (PBS), penicillin (2×106 U/L), streptomycin (200 mg/L), and amphotericin B (250 mg/L) (antimicrobial drugs from Lonza, Walkersville, MD, USA). Samples were chilled on ice until delivered to the laboratory (within 24 hours). All samples were stored at –80°C until used.

Acknowledgment

This work was funded by the US Department of Health and Human Services, National Institutes of Health, National Institute of Allergy and Infectious Diseases, under contract no. HHSN266200700005C, and supported by the American Lebanese Syrian Associated Charities.

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Figures

• Figure 1. . Location of surveillance governorates and percentage of avian influenza virus detection in each governorate, Egypt, 2010–2012.
• Figure 2. . Avian influenza virus infections, by month, Egypt, 2010–2012. Blue bars, detection of the virus in birds; red dots, cases of influenza A(H5N1) virus infections in humans; and black...
• Figure 3. . Subtypes of influenza A viruses detected in poultry, by month, by using reverse transcription PCR, Egypt, 2010–2012.
• Figure 4. . Phylogenetic tree of the hemagglutinin gene of influenza A(H5N1) viruses from Egypt, 2010–2012. Scale bar indicates phylogenetic distance (1 base substitution/100 positions).
• Figure 5. . Phylogenetic tree of the hemagglutinin gene of influenza A(H9N2) viruses from Egypt, 2010–2012. Scale bar indicates phylogenetic distance (1 base substitution/100 positions).

Tables

• Table 1. Primers used for H5 and H9 subtyping of avian influenza viruses from Egypt, 2010–2012
• Table 2. Epizootiologic data for avian influenza virus isolated from poultry in 7 governorates in Egypt, 2010–2012
• Table 3. Epizootiologic data for avian influenza virus subtypes H5N1, H9N2, and H5/H9 , Egypt, 2010–2012

Technical Appendix

• Technical Appendix. . Avian influenza A virus detection in poultry in Egypt, by month and by subtype; experimental growth of avian influenza A virus in specific pathogen–free embryonated chicken eggs and... 461 KB 

Suggested citation for this article: Kayali G, Kandeil A, El-Shesheny R, Kayed AS, Gomaa MM, Maatouq AM, et al. Active surveillance for avian influenza virus, Egypt, 2010–2012. Emerg Infect Dis [Internet]. 2014 Apr [date cited]. http://dx.doi.org/10.3201/eid2004.131295

DOI: 10.3201/eid2004.131295