Protective effects of immersion immunization of koi with Escherichia coli DH5α carrying DNA vaccine against koi herpesvirus

This study aims to immunize the koi fingerlings immersion using the formalin-killed and freeze-dried E. coli DH5α carrying plasmid for the KHV DNA vaccine. 200 fish on each tank in a total water volume of 20 L. Each tanks consists of different vaccination group: PBS as control (10 mL; C), empty E. coli DHα (10 mL at 108 CFU mL-1; E), formalinkilled E. coli DHα:ORF81 (10 mL at 108 CFU mL-1; KE), freezedried E.coli DHα:ORF81 without formalin inactivation (10 mL at 108 CFU mL-1; FE), and formalin-killed and then freeze-dried E. coli DHα:ORF81 (10 mL at 108 CFU mL-1; KFE). The bath vaccination was conducted for 1 × 30 min. For the challenged test, fish were immersed with the 100 mL of LD50 dose of KHV (10–2 dilution from the KHV stock) for 30 min. The vaccination using E. coli DH5α:ORF81 could significantly modulate the innate and adaptive immunity of the fish and result in higher fish survival after KHV infection. The vaccination using formalin-killed or formalin-killed and freezedried E. coli DH5α:ORF81 will be further developed as an alternative to the costefficient koi or carp fingerlings vaccination method.