Development and Efficacy Evaluation of an SP01-adjuvanted Inactivated Escherichia Coli Mutant Vaccine Against Bovine Coliform Mastitis

Escherichia coli (E. coli) is one of the most common pathogens causing clinical mastitis in cattle, but no vaccine is available to prevent this disease in China. Therefore, development of an E. coli vaccine against bovine clinical mastitis is urgently needed. The candidate vaccine (Ch-O111-1) and challenge (LZ06) strains were screened from milk samples of cows with clinical mastitis. To extend the cross-protection of the Ch-O111-1 strain, we deleted the galE gene fragment of the Ch-O111-1 strain through homologous recombination between the Ch-O111-1 strain and pCVD442/ΔgalE plasmid, which was identified through conventional methods, including PCR, SDS-PAGE and sequencing. The Ch-O111-1/ΔgalE (Z9) strain was characterized by extensive cross-reactivity and attenuated virulence. We prepared inactivated Z9 vaccines with different adjuvants. Immunization of inactivated Z9 antigen induced adjuvant-, dosage- and inoculation time-dependent antibody titers in cows and mice. Furthermore, immunization with SP01-adjuvanted inactivated Z9 vaccine protected cows against severe clinical mastitis caused by LZ06 and protected mice against death due to LZ06. An SP01-adjuvanted inactivated Z9 vaccine was successfully developed and found to protect cows against severe mastitis caused by Escherichia coli.