Evaluation of the U.S. Department of Agriculture's egg pasteurization processes on the inactivation of high-pathogenicity avian influenza virus and velogenic Newcastle disease virus in processed egg products.

Globally, 230,662 metric tons of liquid egg products are marketed each year. The presence of highly pathogenic avian influenza (HPAI) or Newcastle disease in an exporting country can legitimately inhibit trade in eggs and processed egg products; development and validation of pasteurization parameters are essential for safe trade to continue. The HPAI virus (HPAIV) A/chicken/Pennsylvania/1370/1983 (H5N2) and velogenic Newcastle disease virus (vNDV) AMPV-1/chicken/California/S01212676/2002 were inoculated into five egg products and heat treated at various times and temperatures to determine thermal inactivation rates to effect a 5-log viral reduction. For HPAIV and vNDV, the pasteurization processes for fortified, sugared, plain, and salted egg yolk, and homogenized whole egg (HPAIV only) products resulted in >5-log reductions in virus at the lower temperature-longer times of U.S. Department of Agriculture (USDA)-approved Salmonella pasteurization processes. In addition, a >5-log reduction of HPAIV was also demonstrated for the five products at the higher temperatures-shorter times of USDA-approved pasteurization processes, whereas the vNDV virus was adequately inactivated in only fortified and plain egg yolk products. For the salted and sugared egg yolk products, an additional 0.65 and 1.6 min of treatment, respectively, at 63.3 °C was necessary to inactivate 5 log of vNDV. Egg substitute with fat does not have standard USDA pasteurization criteria, but the D59-value was 0.75 min, adequate to inactivate 5 log of vNDV in <4 min.

Reduction of high pathogenicity avian influenza virus in eggs from chickens once or twice vaccinated with an oil-emulsified inactivated H5 avian influenza vaccine.

The negative impact of high pathogenicity avian influenza virus (HPAIV) infection on egg production and deposition of virus in eggs, as well as any protective effect of vaccination, is unknown. Individually housed non-vaccinated, sham-vaccinated and inactivated H5N9 vaccinated Once or Twice adult White leghorn hens were challenged intranasally/intratracheally 3-weeks post-vaccination with H5N2 HPAIV. The non-/sham-vaccinated layers experienced 100% mortality (0% survivability) within 3-4 days post-challenge (DPC), and major changes to reproductive parameters including precipitous drops in egg production (79-0% in <5 days), production of soft and thin-shelled eggs, and deposition of virus in albumin and yolk, and on the egg shell surface of 53% of eggs. By comparison, the three H5-vaccinated groups had 83%, 100% and 100% survivability after challenge; the two H5-vaccinated Once hens that died had low pre-challenge HI titers (GMT=16). H5-vaccinated Once or Twice groups maintained egg production after challenge (63%), but there was a mild and significant reduction in egg production as compared to pre-challenge egg production (79%). H5-vaccinated groups had reduced number of virus contaminated eggs (28%), and in most groups, reduced quantity of virus in contaminated eggs compared to non-/sham-vaccinated groups. No HPAIV-positive eggs were laid on or after 5 DPC. In conclusion, HPAIV infection had major negative impact on egg production and other reproductive parameters. H5-vaccination Once or Twice prevented declines in egg production after HPAIV challenge, reduced number of virus-infected eggs, and typically reduced the titer of virus in internal contents and on eggshell surface.

Thermal inactivation of avian influenza virus and Newcastle disease virus in a fat-free egg product.

High-pathogenicity avian influenza (HPAI) virus, low-pathogenicity avian influenza (LPAI) virus, virulent Newcastle disease virus (vNDV) and low-virulent Newcastle disease virus (lNDV) can be present on the eggshell surface, and HPAI viruses and vNDV can be present in the internal contents of chicken eggs laid by infected hens. With the increase in global trade, egg products could present potential biosecurity problems and affect international trade in liquid and dried egg products. Therefore, the generation of survival curves to determine decimal reduction times (D(T)-values) and change in heat resistance of the viruses (z(D)-value) within fat-free egg product could provide valuable information for development of risk reduction strategies. Thermal inactivation studies using A/chicken/Pennsylvania/1370/83 (H5N2) HPAI virus resulted in D(55)-, D(56)-, D(56.7)-, D(57)-, D(58)-, and D(59)-values of 18.6, 8.5, 3.6, 2.5, 0.4, and 0.4 min, respectively. The z(D)-value was 4.4 °C. LPAI virus A/chicken/New York/13142/94 (H7N2) had D(55)-, D(56.7)-, D(57)-, D(58)-, D(59)-, and D(60)-values of 2.9, 1.4, 0.8, 0.7, 0.7, and 0.5 min, respectively, and a z-value of 0.4 °C. vNDV avian paramyxoviruses of serotype 1 (AMPV-1)/chicken/California/212676/2002 had D(55)-, D(56)-, D(56.7)-, D(57)-, D(58)-, and D(59)-values of 12.4, 9.3, 6.2, 5, 3.7, and 1.7 min, respectively. The z(D)-value was 4.7 °C. lNDV AMPV-1/chicken/United States/B1/1948 had D(55)-, D(57)-, D(58)-, D(59)-, D(61)-, and D(63)-values of 5.3, 2.2, 1.1, 0.55, 0.19, and 0.17 min, respectively, and a z(D)-value of 1.0 °C. Use of these data in developing egg pasteurization standards for AI and NDV-infected countries should allow safer trade in liquid egg products.

Susceptibility of North American ducks and gulls to H5N1 highly pathogenic avian influenza viruses.

Since 2002, H5N1 highly pathogenic avian influenza (HPA1) viruses have been associated with deaths in numerous wild avian species throughout Eurasia. We assessed the clinical response and extent and duration of viral shedding in 5 species of North American ducks and laughing gulls (Larus atricilla) after intranasal challenge with 2 Asian H5N1 HPAI viruses. Birds were challenged at approximately equal to 10 to 16 weeks of age, consistent with temporal peaks in virus prevalence and fall migration. All species were infected, but only wood ducks (Aix sponsa) and laughing gulls exhibited illness or died. Viral titers were higher in oropharyngeal swabs than in cloacal swabs. Duration of viral shedding (1-10 days) increased with severity of clinical disease. Both the hemagglutination-inhibition (HI) and agar gel precipitin (AGP) tests were able to detect postinoculation antibodies in surviving wood ducks and laughing gulls; the HI test was more sensitive than the AGP in the remaining 4 species.

Experimental study to determine if low-pathogenicity and high-pathogenicity avian influenza viruses can be present in chicken breast and thigh meat following intranasal virus inoculation.

Two low-pathogenicity (LP) and two high-pathogenicity (HP) avian influenza (AI) viruses were inoculated into chickens by the intranasal route to determine the presence of the AI virus in breast and thigh meat as well as any potential role that meat could fill as a transmission vehicle. The LPAI viruses caused localized virus infections in respiratory and gastrointestinal (GI) tracts. Virus was not detected in blood, bone marrow, or breast and thigh meat, and feeding breast and thigh meat from virus-infected birds did not transmit the virus. In contrast to the two LPAI viruses, A/chicken/Pennsylvania/1370/1983 (H5N2) HPAI virus caused respiratory and GI tract infections with systemic spread, and virus was detected in blood, bone marrow, and breast and thigh meat. Feeding breast or thigh meat from HPAI (H5N2) virus-infected chickens to other chickens did not transmit the infection. However, A/lchicken/Korea/ES/2003 (H5N1) HPAI virus produced high titers of virus in the breast meat, and feeding breast meat from these infected chickens to other chickens resulted in Al virus infection and death. Usage of either recombinant fowlpox vaccine with H5 AI gene insert or inactivated Al whole-virus vaccines prevented HPAI virus in breast meat. These data indicate that the potential for LPAI virus appearing in meat of infected chickens is negligible, while the potential for having HPAI virus in meat from infected chickens is high, but proper usage of vaccines can prevent HPAI virus from being present in meat.

Heat inactivation of avian influenza and Newcastle disease viruses in egg products.

Avian influenza (AI) and Newcastle disease (ND) viruses are heat labile viruses, but exact parameters for heat inactivation at egg pasteurization temperatures have not been established. In this study we artificially infected four egg products with two AI (one low [LP] and one high pathogenicity [HP]) and three ND (two low and one highly virulent) viruses, and determined inactivation curves at 55, 57, 59, 61 and 63 degrees C. Based on D(t) values, the time to inactivation of the viruses was dependent on virus strain and egg product, and was directly related to virus titre, but inversely related to temperature. For all temperatures, the five viruses had the most rapid and complete inactivation in 10% salt yolk, while the most resistant to inactivation was HPAI virus in dried egg white. This study demonstrated that the LPAI and all ND viruses were inactivated in all egg products when treated using industry standard pasteurization protocols. By contrast, the HPAI virus was inactivated in liquid egg products but not in dried egg whites when using the low-temperature industry pasteurization protocol.

Domestic poultry and SARS coronavirus, southern China.

SARS coronavirus injected intratracheally into chickens, turkeys, geese, ducks, and quail, or into the allantoic sac of their embryonating eggs, failed to cause disease or replicate. This finding suggests that domestic poultry were unlikely to have been the reservoir, or associated with dissemination, of SARS coronavirus in the animal markets of southern China.