Masood RABBANI, Arfan AHMAD, Khushi MUHAMMAD, Asfa RASOOL, Sameera AKHTAR, Aftab Ahmed ANJUM, Javed MUHAMMAD, Ali Ahmad SHEIKH, Fakhra LIAQAT, Fariha AKHTAR, Rabia TAHIR

Comparative evaluation of factors affecting hemagglutinatingactivity of avian influenza (H9) virus

Influenza virus H9 was standardized for hemagglutination assay using different factors such as red blood cell (RBC) types, concentrations, diluent types, and storage times. Avian influenza virus H9 was grown in embryonated chicken eggs and confirmed by spot agglutination. A significant (P < 0.05) difference with the highest mean titer (9.00 ± 0.00) was observed using RBCs from different species. Nonsignificant differences (P > 0.05) were found between human blood type O, chicken, and dog RBCs, as well as among rabbit, pigeon, sheep, and parrot. The highest titer (9.00 ± 0.00) with a nonsignificant difference was found using virus stored at 4 °C and 20 °C while 37 °C showed the lowest significant mean hemagglutinin (HA) titer (11.08 ± 188.21). Nonsignificant differences were observed in HA titers against H9 virus stored for 3, 4, 5, and 6 months. Nonsignificant differences were found between the use of normal saline and 0.5% peptone water with the lowest HA titers of 7.83 ± 0.40 and 8.00 ± 0.00, respectively, while the highest HA titer (9.00 ± 0.00) with nonsignificant difference was observed by using HA-HI buffer and phosphate buffer saline as diluents. RBCs with 0.5% and 1% concentrations showed nonsignificant difference in HA titer but significant difference with 0.1% RBCs.

PDF

___

1. Van Reeth K. Avian and swine influenza viruses: our current understanding of the zoonotic risk. Vet Res 2007; 38: 243260.
2. Tsukamoto K, Ashizawa T, Nakanishi K, Kaji N, Suzuki K, Shishido M, Okamatsu M, Mase M. Use of reverse transcriptase PCR to subtype N1 to N9 neuraminidase genes of avian influenza viruses. J Clin Microbiol 2009; 47: 23012303.
3. Gamblin SJ, Skehel JJ. Influenza hemagglutinin and neuraminidase membrane glycoproteins. J Biol Chem 2010; 285: 2840328409.
4. Naeem K, Hussain M. An outbreak of avian Influenza in poultry in Pakistan. Vet Rec 1995; 137: 439.
5. Naeem K, Ullah A, Manvell RJ, Alexander DJ. Avian influenza A subtype H9N2 in poultry in Pakistan. Vet Rec 1999; 145: 560.
6. Allan WH, Lancaster JE, Toth B. Newcastle Disease Vaccines, Their Production and Use. Rome, Italy: Food and Agriculture Organization of the United Nations; 1978.
7. Beard CW. Avian influenza antibody detection by immunodiffusion. Avian Dis 1970; 14: 337341.
8. Witthawat, Weena WP, Kranapone P, Pirom P, Kridsda C. Seroprevalence of H PA I H 5N 1 virus in domestic dogs and cats living in repeated outbreak areas in Thailand. Bangkok, Thailand: Thai Ministry of Public Health; 2009.
9. Hussain M, Mehmood M, Ahmad A, Shabbir M, Yaqub T. Factors affecting hemagglutination activity of avian influenza virus subtype H5. J Vet Anim Sci 2008; 1: 3136.
10. Pawar SD, Kale SD, Rawankar AS, Koratkar SS, Raut CG, Pande SA, Mullick J, Mishra AC. Avian influenza surveillance reveals presence of low pathogenic avian influenza viruses in poultry during 2009-2011 in the West Bengal State, India. Virol J 2012; 9: 151.
11. Kurskaia O, Durymanov A, Sharshov K, Zaĭkovskaia A, Shestopalov A, Ilicheva T. Hemagglutination inhibition test for retrospective diagnosis of avian influenza in mammals. Zh Mikrobiol Epidemiol Immunobiol 2009; 75.
12. Burnet F, Edney M. Influence of ions on the interaction of influenza virus and cellular receptors or soluble inhibitors of haemagglutination. Aust J Exp Biol Med Sci 1952; 30: 105118.
13. Epand RM, Epand RF. Thermal denaturation of Influenza virus and its relationship to membrane fusion. J Biochem 2002; 1: 841848.
14. Stallknecht DE, Kearney MT, Shane SM, Zwank PJ. Effects of pH, temperature, and salinity on persistence of avian influenza viruses in water. Avian Dis 1991; 34: 412418.
15. Stallknecht DE, Shane SM, Kearney MT, Zwank PJ. Persistence of avian influenza viruses in water. Avian Dis 1990; 34: 406411.
16. Shahid MA, Abubakar M, Hameed S, Hassan S. Avian influenza virus (H5N1); effects of physico-chemical factors on its survival. Virol J 2009; 6: 3844.