TITLE:
Immunomodulatory Effect of Purified Exotoxins of Staphylococcus aureus in Association with Bird Flu Virus Vaccine in Broilers
AUTHORS:
Muhammad Usman Ghani, Muhammad Danish Mehmood, Amna Javed, Farheen Ansari, Huma Anwar, Sana Noreen, Sajjad Hussain Shah
KEYWORDS:
AIV H9N2, Staphylococcus aureus Toxoid, Super Antigen, Immunomodulation, HI Assay
JOURNAL NAME:
American Journal of Molecular Biology,
Vol.9 No.1,
November
27,
2018
ABSTRACT: Immunization is the most effective method
still used against infectious agents. Although not always, vaccines
ineffectiveness is reported enormously against
many of the pathogens throughout the world in poultry, particularly in case of killed or sub unit vaccine. The current project
is, therefore, carried out as a preliminary study on broiler chickens to
investigate the modulation of immune system against avian influenza virus in
association with purified Staphylococcus aureus toxoid. After isolation
of Gram positive cocci bacteria on mannitol salt agar from raw milk, yogurt and
chicken meat were subsequently biochemically characterized by using rapid
diagnostic kit. Pure culture of S. aureus was inoculated into digitally controlled bio-fermentor containing mannitol salt broth for production of toxins.
Enormous production of bacteria was passed through sequence of filtration
system based on 0.45 μm followed by 0.22 μm size. The centrifugation of the
filtrate was made at 10,000 rpm for 60 minutes at 5℃ followed by 56,100 rpm
for 20 minutes and clear supernatant containing Staphylococcus enterotoxin
(SEs) was obtained. Bradford estimation of proteins further provided 305 μg/ml
of SEs toxoid. Four types of oil adjuvant avian influenza type H9N2 virus
vaccines (without toxoid, 91.5 μg/0.3ml, 22.8 μg/0.3ml and 11.43 μg/0.3ml) were injected into healthy AI H9N2 susceptible broilers and anti-H9N2 HI
antibody titers were measured in terms of hemagglutination inhibition test. It
was observed that on the 8th day, post vaccination cumulative mean anti AIH9 HI antibody titer of G-1, G-2, G-3, G-4 and G-5 was 3.13 ± 0.406, 5.13 ± 0.246, 3.96 ± 0.159,
3.25 ± 0.237 and 0.78 ± 0.467 respectively. It was found that all the vaccines
induced protective titers 18 days’ post vaccination, but vaccine containing 91.5 μg/dose of SEs toxoid showed significantly higher (P , particularly in inactivated vaccines, could be the better choice for modulation of immediate and better immune
response in future vaccines technologies.