Three cohorts of juvenile and subadult Chinook salmon Oncorhynchus tshawytscha received multiple treatments with macrolide antibiotics for bacterial kidney disease (BKD) during rearing in a captive broodstock program. A total of 77 mortalities among the cohorts were screened for Renibacterium salmoninarum, the etiologic agent of BKD, by agar culture from kidney, and isolates from 7 fish were suitable for growth testing in the presence of macrolide antibiotics. The minimum inhibitory concentration (MIC) of erythromycin and azithromycin was determined by a modification of the standardized broth assay using defined medium. The American Type Culture Collection (ATCC) type strain 33209 exhibited a MIC of 0.008 µg ml -1 to either erythromycin or azithromycin. Isolates from 3 fish displayed MICs identical to the MICs for the ATCC type strain 33209. In contrast, isolates from 4 fish exhibited higher MICs, ranging between 0.125 and 0.250 µg ml -1 for erythromycin and between 0.016 and 0.031 µg ml -1 for azithromycin. Sequence analysis of the mutational hotspots for macrolide resistance in the 23S rDNA gene and the open reading frames of ribosomal proteins L4 and L22 found identical sequences among all isolates, indicating that the phenotype was not due to mutations associated with the drug-binding site of 23S rRNA. These results are the first report of R. salmoninarum with reduced susceptibility to macrolide antibiotics isolated from fish receiving multiple antibiotic treatments.
KEY WORDS:Antimicrobial agent susceptibility testing · Macrolide antibiotic · Reduced susceptibility · Renibacterium salmoninarum · Bacterial kidney disease
Resale or republication not permitted without written consent of the publisherDis Aquat Org 80: [173][174][175][176][177][178][179][180] 2008 tible to infection throughout the entire life cycle. For fish culturists, the windows of opportunity to deliver therapeutics against BKD are limited, and antibiotics are the most commonly used agents.By 1990, erythromycin was demonstrated to be potentially effective against vertical transmission of Renibacterium salmoninarum (Bullock & Leek 1986, Evelyn et al. 1986, Brown et al. 1990, and injection of female broodstock with erythromycin is currently a routine procedure in many US rearing facilities (e.g. Washington Department of Fish and Wildlife 2002, US Fish andWildlife Service 2005). In the US, erythromycin may be used in food salmonids under an Investigational New Animal Drug (INAD) application, although approval is still pending (National Coordinator for Aquaculture New Animal Drug Applications 2007). In captive broodstock programs for endangered stocks of salmon, a percentage of fish are held through the entire life cycle, and the risk of BKD epidemics can be high. In those programs oral or injected macrolides may be administered anytime during growth to maturity.Azithromycin, a semi-synthetic derivative of erythromycin, has a broader spectrum of activity against bacteria than erythromycin, and it is active against Grampositive bact...