Recombinant IBV (B/Florida/4/2006) Hemagglutinin (a.a. 362-555) [hFc]

Avibacterium paragallinarum is an important pathogen affecting the respiratory tract of chickens. There is a paucity of information on the molecular characteristics and pathology of A. paragallinarum in Turkish poultry flocks. In the present study, broiler and layer flocks (n = 2) suspected of viral infections with serious respiratory signs and significant mortality were visited. Chickens showed various disease signs and necropsy lesions, including purulent nasal discharge, respiratory distress, facial edema, sticky eyes, mucoid tracheitis, hemorrhagic inflammation of the infraorbital sinuses along with fibrinous mass and conjunctivitis. Histopathological lesions included loss of cilia along with necrosis and exfoliation of the superficial mucosal epithelium of the trachea, facial cellulitis, dermatitis, fibrinous plasmatic edema and infiltration of inflammatory cells, especially heterophils. A. paragallinarum was detected in tissue samples by species-specific polymerase chain reaction (PCR). The sequencing and phylogenetic analysis of the core region of the hemagglutinin (HA) gene revealed that Turkish strains detected here belonged to serotype A (serovar A1). They were related to strains reported from India (VRDC), the United States (0083), and Japan (0221), which are representatives of serovar A1. A homology of 88-90% was found between Indian strains and the Turkish strains detected in this study. Surprisingly, only vaccine strains of infectious bronchitis virus (IBV) were detected as a co-infecting agent in all samples positive for A. paragallinarum. Our findings suggest that A. paragallinarum may be an emerging pathogen in Turkish poultry flocks, and direct PCR may facilitate rapid diagnosis of infectious coryza. These results will also help to develop control strategies for A. paragallinarum.

The defense against influenza virus (IV) infections still poses a series of challenges. The current antiviral arsenal against influenza viruses is in fact limited; therefore, the development of new anti-influenza strategies effective against antigenically different viruses is an urgent priority. Bioactive compounds derived from medicinal plants and fruits may provide a natural source of candidates for such broad-spectrum antivirals. In this regard, cranberry (Vaccinium macrocarpon Aiton) extracts on the basis of their recognized anti-adhesive activities against bacteria, may provide potential compounds able to prevent viral attachment to target cells. Nevertheless, only few studies have so far investigated the possible use of cranberry extracts as an antiviral tool. This study focuses on the suitability of a cranberry extract as a direct-acting anti-influenza compound. We show that the novel cranberry extract Oximacro (R) inhibits influenza A and B viruses (IAV, IBV) replication in vitro because of its high content of A-type proanthocyanidins (PAC-A) dimers and trimers. Mechanistic studies revealed that Oximacro (R) prevents attachment and entry of IAV and IBV into target cells and exerts a virucidal activity. Oximacro (R) was observed to interact with the ectodomain of viral hemagglutinin (HA) glycoprotein, thus suggesting the interference with HA functions and a consequent loss of infectivity of IV particles. Fluorescence spectroscopy revealed a reduction in the intrinsic fluorescence of HA protein after incubation with purified dimeric PAC-A (PAC-A2), thus confirming a direct interaction between HA and Oximacro (R) PAC-A2. In silico docking simulations further supported the in vitro results and indicated that among the different components of the Oximacro (R) chemical profile, PAC-A2 exhibited the best binding propensity with an affinity below 10 nM. The role of PAC-A2 in the anti-IV activity of Oximacro (R) was eventually confirmed by the observation that it prevented IAV and IVB replication and caused the loss of infectivity of IV particles, thus indicating PAC-A2 as the major active component of Oximacro (R). As a whole, these results suggest Oximacro (R) as a potential candidate to create novel antiviral agents of natural origin for the prevention of IV infections.