DAO 72:135-146 (2006)  -  doi:10.3354/dao072135

Aquatic Francisella-like bacterium associated with mortality of intensively cultured hybrid striped bass Morone chrysops × M. saxatilis

V. E. Ostland1,*, J. A. Stannard1, J. J. Creek1, R. P. Hedrick2, H. W. Ferguson3, J. M. Carlberg1, M. E. Westerman1

1Kent SeaTech Corporation, 11125 Flintkote Ave., Suite J, San Diego, California 92121, USA
2Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California 95616, USA
3Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, UK

ABSTRACT: The present study identifies an emerging disease associated with an aquatic Francisella-like bacterium that can cause mortality in hybrid striped bass Morone chrysops × M. saxatilis reared intensively in freshwater. Clinically affected fish were lethargic, had scattered haemorrhagic cutaneous lesions and diffuse gill pallor. The head kidney and spleen were markedly swollen and contained numerous interstitial granulomas; histological examination revealed small, pleomorphic Gram-negative coccobacilli within vacuolated cells. The bacterium could not be cultured from head kidney homogenates either with standard or enriched microbiological media or following inoculation of a Chinook salmon embryo (CHSE)-214 cell line. No amplification product was obtained from head kidney DNA by polymerase chain reaction (PCR) assay using Piscirickettsia salmonis-specific primers. PCR analysis of infected head kidney homogenate with primers designed for the eubacterial 16S rRNA produced a single amplicon. Phylogenetic analysis of this DNA sequence demonstrated that the sequence aligned most closely with members of the genus Francisella, identified from tilapia Oreochromis spp. in Taiwan and an aquatic Francisella species that was recently isolated from the three-line grunt Parapristipoma trilineatum in Japan. This Francisella-like disease was transmitted to naïve hybrid striped bass fingerlings by intraperitoneal injection of tissue homogenates prepared from a natural outbreak. All fish developed gross and histological lesions identical to those from natural outbreaks. Intracellular Gram-negative bacteria were observed within the cytoplasm of cells (presumably macrophages) within the granulomas, but bacteria were not recovered. The 16S DNA sequence of the bacterium obtained from tissues of experimentally infected fish was identical to that obtained from the fish used as infected donor tissue.


KEY WORDS: Francisella-like bacterium · Intracellular bacterial disease · Hybrid striped bass · Emergent disease in aquaculture