Abstract
The role of reactive oxygen species (ROS) in the killing of the bacterial pathogen Aeromonas salmonicida by in vitro cultured rainbow trout (Oncorhynchus mykiss) head kidney macrophages has been investigated. Using a microtitre plate bactericidal assay the respiratory burst pathway responsible for ROS generation was inhibited completely using the NADPH oxidase inhibitor trifluoperazine and partially by the use of ROS scavengers (formate and/or mannitol) and the antioxidative enzymes SOD and/or catalase. Increased bacterial survival of an avirulent strain (MT004) in the presence of trifluoperazine, catalase and formate confirmed that ROS such as hydrogen peroxide and hydroxyl radicals were indeed an important killing agent. This was further confirmed using sodium nitroprusside and aminotriazole, inhibitors of SOD and catalase respectively. The presence of aminotriazole caused a significantly increased bactericidal activity against a virulent strain (MT423) normally resistant to killing by cultured resident rainbow trout head kidney macrophages.
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