Although the role of glutamate was diminished in this study, it contributed substantially to fighting against apoptosis. In summary, GSE-induced apoptosis was mediated by mitochondria-dependent pathway. Although initiated by the mitochondria, other organelles were also involved in this apoptosis. The changes in the yeast cells during apoptosis were indicative of their struggle with cell death. However, the destruction of the mitochondrial 60 S ribosomal protein L14-A and the Tigecycline cessation of the conversion of pantothenic acid to CoA resulted in apoptosis. The method can be used as a preliminary study of apoptosis, to identify interest proteins or metabolites and then further detailed study is needed to find out the exact mechanism of apoptosis. The predicted amino acid sequence analysis of the NA and matrix genes revealed that the virus isolates are likely to be sensitive to commonly used influenza drugs such as amantadine and oseltamivir. The E627K mutation in the PB2 protein, associated with increased virulence of influenza A H5N1 viruses in 10-Gingerol mammals was not present. Deletion of five amino acids in the N-terminal of the NS1 protein from position 80 to 84 which might enhance cytokine expression by macrophages is similar to other viruses. Molecular markers for possible increase in the virulence of this virus in chicken and mice and also inhibition of host immune responses. The pathogenicity of viruses in mice needs to be ascertained to assess their possible public health implications. The intravenous pathogenicity index calculated for all the seven isolates ranged from indicating high pathogenicity to chickens. Further investigations are needed to ascertain whether the presence of concurrent infection with other infectious agents such as duck viral enteritis or duck viral hepatitis contributed to the increased mortality. In chickens, even though 100% mortality was recorded in one shed, no mortality was observed in other sheds indicating that strict biosecurity measures prevented onward spread. Interestingly, no mortality was observed in other poultry within 3 KM radius of the first outbreak in the one month that separated the two outbreaks and all the samples collected during the intensive post-outbreak surveillance program were negative for avian influenza. Hence, there is a possibility that the introduction to the Gandhigram farm was not through direct transmission from the index farm. The role of fomites or local migratory birds needs to be considered. The most probable route of this transmission could be the movement of land based poultry or local migratory birds. Following the repeated detection H5N1 clade 2.3.2 viruses in wild birds in Hong Kong in 2006�C2008 and in waterfowl and poultry in Russia and Japan, we suggested that this virus clade may have become endemic in wild birds and may be spreading via long distance bird migration. The possible dissemination of influenza A H5N1 throughout Eurasia through wild migratory birds has been previously discussed. The emergence and spread of H5N1 virus in South Asian region supports the contention that this virus clade is probably established in wild birds and land based poultry and is spreading its geographical range just as clade 2.2 before it. The WHO report on antigenic and genetic characteristics of zoonotic influenza viruses and development of candidate vaccine viruses for pandemic preparedness also reveals isolation of clade 2.3.4.2 viruses in Myanmar and Bangladesh which showed reduced reactivity with post-infection ferret antisera against the clade 2.3.4 viruses. Continued circulation of the H5N1 viruses of various subclades which are more adapted to land based poultry in a highly populated region might lead to evolution of pandemic strains with devastating consequences.