Molecular characterization of Toll-like receptor type-3 in mallard duck and its response to Newcastle disease virus infection.

Toll-like receptors (TLRs), type I transmembrane pattern recognition receptors (PRRs), are composed of the extracellular domain that is implicated in the recognition of microbial products and initiates the innate and adaptive immune response. Previous reports on TLRs in birds showed significant levels of inter- and intraspecific genetic variation. Little is known about the structure and function of the avian immune system, especially waterfowl species. This work aimed to identify and clone Anas platyrhynchos (mallard duck) TLR-3 (dTLR-3) and its expression level following challenge with velogenic Newcastle disease virus (NDV) as a model for waterfowl species. The mallard duck TLR-3 full-length cDNA sequence had been cloned, which consisted of 2457 nucleotides. The translated amino acid sequence showed identity degree as 97% with Muscovy duck, 95% with geese, 89% with helmeted guineafowls, 88% with the chickens TLR-3 gene, 82% with turkey TLR-3, and 79% with zebra finch, while it showed 54% with human one; the analysis data suggested that the new sequence is probably homologous to vertebrates' TLR-3. The predicted protein encoded by the duck dTLR-3 mRNA sequence is composed of 819 amino acids. Analysis of the deduced amino acid sequence indicated that dTLR-3 has typical structural features and contains the main components of proteins in the TLR family. The dTLR-3 expressed in almost all examined tissues of mallard duck following quantitative real-time polymerase chain reaction (qPCR) analysis and using B-actin as a housekeeping gene. To check the functionality of the receptor and its role in viral infection, we evaluate the expression level in different tissues and its changes following NDV infection. The results showed significant (P < 0.05) upregulated in the brain at 24 h (1.84-fold), reached a peak at 48 h (4.82-fold), and recovered to normal levels at 72 h post-infection. These results indicate a complete and functional dTLR-3 that is orthologous to other vertebrate receptors with its potential role in early response against viral infection in mallard duck species.

Comparison of the Effectiveness of Two Different Vaccination Regimes for Avian Influenza H9N2 in Broiler Chicken.

Low pathogenic avian influenza virus is one of the major threats that has been affecting the poultry industry in the Middle East region for decades. Attempts to eradicate this disease have failed. Currently, there are commercial vaccines that are either imported or produced locally from recently circulating isolates of H9N2 in Egypt and Middle Eastern countries. This present work focused on comparing the effectiveness of two vaccines belonging to these categories in Egypt. Two commercial broiler flocks (Cobb-500 Broiler) with maternally derived immunity (MDA) against H9N2 virus were employed and placed under normal commercial field conditions or laboratory conditions. Immunity was evaluated on the basis of detectable humoral antibodies against influenza H9N2 virus, and challenge was conducted at 28 days of life using a recent wild H9N2 virus. The results showed that vaccination on the 7th day of life provided significantly higher immune response in both vaccine types, with significantly lower virus shedding compared to vaccination at day 1 of life, regardless of field or laboratory conditions. In addition, the vaccine produced from a recent local H9N2 isolate (MEFLUVAC-H9-16) provided a significantly higher humoral immune response under both field and laboratory conditions, as measured by serology and virus shedding (number of shedders and amount of shedding virus), being significantly lower following challenge on the 28th day of life, contrary to the imported H9 vaccine. In conclusion, use of H9N2 vaccine at 7 days of life provided a significantly higher protection than vaccination at day 1 of life in birds with MDA, suggesting vaccination regimes between 5-8-days of life for broiler chicks with MDA. Moreover, use of a vaccine prepared from a recently circulating H9N2 virus showed significantly higher protection and was more suitable for birds in the Middle East.