Isolation of Avian Influenza Virus A Subtype H5N1 from Internal Contents (Albumen and Allantoic Fluid) of Japanese Quail (Coturnix coturnix japonica) Eggs and Oviduct during a Natural Outbreak

Abstract

Avian influenza virus (AIV) was recovered from the internal contents of eggs, including mixture of albumen and allantoic fluid, and from the oviduct of naturally infected Japanese quail (Coturnix coturnix japonica) flocks in the southern part of Thailand. The virus titers of 10(4.6)-10(6.2) ELD(50)/mL were directly measured from the internal content of infected eggs. The virus was isolated by chorioallantoic sac inoculation of embryonating chicken eggs. Infected allantoic fluid was identified as hemagglutinating virus and then was indicated the presence of H5 hemagglutinin. The virus was confirmed to be H5N1 subtype influenza A virus by reverse transcriptase-polymerase chain reaction. Additionally, real-time reverse transcriptase-polymerase chain reaction assay could specifically detect influenza virus subtype H5. Furthermore, indirect fluorescent antibody (IFA) test by using specific anti-influenza A monoclonal antibody indicated that virus antigens were detected in the parenchyma of multiple tissues. Systemic localization of viral antigen detected was certainly considered to be viremic stage. In addition, influenza virus antigen was also detected by IFA in allantoic fluid sediments isolated from internal content of egg or oviduct. The conclusion of isolated AIV type A subtype H5N1 from these two infected materials was correlated to the viremic stage of infection because the virus antigens could be observed in almost all tissues. Conclusively, the need for adequate safeguards to prevent contamination and spread of the virus to the environment during movement of eggs--including hatching eggs, cracked eggs, and other relevant infected materials-- or egg consumption from area of outbreak is emphasized and must not be ignored for the reasons of animal, public, and environmental health.

The onset of virus shedding and clinical signs in chickens infected with high-pathogenicity and low-pathogenicity avian influenza viruses

“…in the gonads of intranasally inoculated chickens (Jones & Swayne, 2004). Asian HPAI H5N1 viruses have been found in the internal contents of quail eggs (Promkuntod et al, 2006).…”

A current review of avian influenza in pigeons and doves (Columbidae)

“…Despite the fact that several experimental studies and reports of natural infections prove that vertical transmission occurs in gallinaceous poultry (Cappucci et al, 1985;Bean et al, 1985;Kilany et al, 2010;Promkuntod et al, 2006;Pillai et al, 2010), there is no evidence to suggest that this occurs in columbids, and no experimental infections to investigate transmission of AIV in pigeon or dove eggs are documented. This is a topic that requires experimental investigation for a conclusion, emphasized by the observation of mild sporadic microscopic lesions in the ovarial thecal epithelial cells of H5N1 inoculated rock pigeons (Brown et al, 2009).…”

Anthropogenic factors and societal response to challenges in the transmission of highly pathogenic avian influenza A (H5N1)

“…Unfortunately, the effects of H5N1 outbreaks for many developing countries were farreaching, affecting food production, poultry export, and the economy. Furthermore, H5N1 virus is detected not only in poultry meat but on eggs (on the shell and within) (Beato et al 2007;Pantin-Jackwood et al 2007;Li et al 2006;Promkuntod, Antarasena, and Prommuang 2006;Beato, Capua, and Alexander 2009), and thus infection poses a risk of spreading and transmission through commodity (Swayne and Beck 2005), especially between regions in developing countries with low capacities to detect the virus.…”