AbstractWe conducted a field trial to evaluate the effectiveness of Aquaflor (50% florfenicol) for controlling mortality associated with Streptococcus iniae in freshwater-reared subadult sunshine bass (female white bass Morone chrysops × male striped bass M. saxatilis). Bacterial samples collected from moribund fish representing a reference population were presumptively identified microbiologically and were later confirmed to be S. iniae by biochemical characterization and polymerase chain reaction. The trial comprised a 1-d acclimation period, 10-d treatment period, and 14-d posttreatment period. During the treatment period, Aquaflor-medicated feed was administered to treated tanks (N = 3) at a target dose of 10 mg of florfenicol·kg of fish−1·d−1, and nonmedicated feed was administered to control tanks (N = 3). At the end of the posttreatment period, mean (±SD) cumulative mortality in treated tanks (9 ± 11%) was significantly (P = 0.040) less than that in control tanks (52 ± 13%). Analysis of medicated feed samples revealed that treated tanks had received an actual dose of 8.3 mg florfenicol·kg fish−1·d−1 (83% of target). No florfenicol was detected in control feed samples. Although the actual florfenicol dose administered to treated tanks was less than the target dose, the trial was accepted by the U.S. Food and Drug Administration Center for Veterinary Medicine as demonstrating the efficacy of Aquaflor to control mortality associated with S. iniae in cultured sunshine bass populations.

01 Feb 2009·Infection and ImmunityQ2 · MEDICINE

Host Immune Response and Acute Disease in a Zebrafish Model ofFrancisellaPathogenesis

Q2 · MEDICINE

Article

Author: Vojtech, Lucia N. ; Ostland, Vaughn ; Hansen, John D. ; Sanders, George E. ; Conway, Carla

ABSTRACTMembers of the bacterial genusFrancisellaare highly virulent and infectious pathogens. New models to studyFrancisellapathogenesis in evolutionarily distinct species are needed to provide comparative insight, as the mechanisms of host resistance and pathogen virulence are not well understood. We took advantage of the recent discovery of a novel species ofFrancisellato establish a zebrafish/Francisellacomparative model of pathogenesis and host immune response. Adult zebrafish were susceptible to acuteFrancisella-induced disease and suffered mortality in a dose-dependent manner. Using immunohistochemical analysis, we localized bacterial antigens primarily to lymphoid tissues and livers of zebrafish following infection by intraperitoneal injection, which corresponded to regions of local cellular necrosis.Francisellasp. bacteria replicated rapidly in these tissues beginning 12 h postinfection, and bacterial titers rose steadily, leveled off, and then decreased by 7 days postinfection. Zebrafish mounted a significant tissue-specific proinflammatory response to infection as measured by the upregulation of interleukin-1β (IL-1β), gamma interferon, and tumor necrosis factor alpha mRNA beginning by 6 h postinfection and persisting for up to 7 days postinfection. In addition, exposure of zebrafish to heat-killed bacteria demonstrated that the significant induction of IL-1β was highly specific to live bacteria. Taken together, the pathology and immune response to acuteFrancisellainfection in zebrafish share many features with those in mammals, highlighting the usefulness of this new model system for addressing both general and specific questions aboutFrancisellahost-pathogen interactions via an evolutionary approach.

01 Apr 2008·Diseases of Aquatic OrganismsQ3 · AGRICULTURAL & FORESTRY SCIENCE

Biochemical, molecular, and virulence characteristics of select Mycobacterium marinum isolates in hybrid striped bass Morone chrysops × M. saxatilis and zebrafish Danio rerio

Q3 · AGRICULTURAL & FORESTRY SCIENCE

Article

Author: Westerman, M. E. ; Watral, V. ; Austin, F. W. ; St-Hilaire, S. ; Ostland, V. E. ; Whipps, C. M. ; Kent, M. L.

A panel of 15 Mycobacterium marinum isolates was characterized by biochemical tests, sequencing the ribosomal DNA intergenic spacer (ITS) region and the heat shock protein 65 gene (hsp65) and pulsed-field gel electrophoresis (PFGE). The biochemical characteristics of all isolates were similar, except for Tween 80 hydrolysis. DNA sequence of hsp65 for a subset of isolates were identical; however, at position 5 of the ITS rDNA, a single nucleotide polymorphism was identified. Isolates possessing a guanine residue at this position (G strains) were unable to hydrolyze Tween 80, while isolates that contained an adenine residue at this position (A strains) were positive for Tween 80 hydrolysis. PFGE successfully discriminated between the G and A strains; all G strains had identical AseI restriction enzyme-cutting patterns while the A strains exhibited a variety of cutting patterns. Eight isolates (4 G and 4 A strains) were further characterized for virulence by experimental infection of hybrid striped bass (HSB) Morone chrysops x M. saxatilis and zebrafish Danio rerio. Seven of the 8 strains produced cumulative mortality ranging from 13.3 to 83.3% in the HSB virulence trial. The M. marinum reference strain ATCC 927T did not produce mortality in HSB. HSB exposed to the G strains had significantly higher cumulative mortality than those exposed to the A strains. When these same isolates were tested in zebrafish, 6 of the 8 strains caused 100% cumulative mortality, with 2 of the A strains being the most pathogenic. In zebrafish, however, ATCC 927T was virulent and produced 28.5% mortality. Collectively, we conclude that the M. marinum G strains are unique and may represent a distinct virulence phenotype in HSB, but this trend was not consistent in zebrafish.

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