Development of sodium polyacrylate nanoparticles based combined mastitis vaccine and assessment of its efficacy in mice model

Abstract

The present study was conducted to develop a stable, safe and effective vaccine against newlinemastitis. For the purpose, two most common mastitis causing bacterial pathogen S. aureus and E. coli newlinewere selected based on previous studies conducted locally and in different part of country. These were newlinecharacterized by cultural, morphological, biochemical tests. Further, these were confirmed by the PCR newlinebased gene amplification. The amplified products were purified and sequenced. The sequences of PCR newlineamplicons have been submitted to NCBI, GenBank to obtain Accession no. MH092071 and KY914488 newlinefor S. aureus and E. coli, respectively. Adjuvants always play critical role in the acceptance and newlinesuccess of any vaccine so well approved and recognized adjuvant MontanideTM GEL 1 was newlineincorporated with virulent S. aureus (MH092071) and E. coli (KY914488) to develop formalized killed newlineadjuvant combined mastitis vaccine PGV (Polymer gel based vaccine) to make it 8.9 X108 newline and 1.85 newlineX108 CFU of formalin killed virulent S. aureus (MH092071) and E. coli (KY914488) in per shot newline(100µl) for mice. The PGV was tested for sterility and found sterile. Its safety was assessed in adult newlineinbred albino female mice and it produced no untoward reaction. For stability testing PGV was stored newlineat different temperature and PGV was found stable for 12 months at refrigeration temperature (4-8°C) newlineand 9 months at room temperature. To assess efficacy of PGV, it was inoculated in 24 adult inbred newlinealbino female mice at the dose of 100µl {8.9 X108 newline and 1.85 X108 CFU of formalin killed virulent S. newlineaureus (MH092071) and E. coli (KY914488)} through subcutaneous route. The control groups were newlineinoculated equal volume of adjuvant and PBS (pH7.4) with similar route. All the mice were challenged newlineon 28th day post vaccination with live virulent S. aureus (MH092071) and E. coli (KY914488) (102 newlineCFU) through intra mammary route and sacrificed on 7 newlineth day post challenge. During experiment, blood newlinewas collected at 7th, 14th, 21st, 28th and 35th days for serum as well as