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Diseases of Aquatic Organisms

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DAO 34:21-26 (1998)  -  doi:10.3354/dao034021

Performance of serum-free broth media for growth of Renibacterium salmoninarum

Clifford E. Starliper1,*, William B. Schill1, Jay Mathias2

1National Fish Health Research Laboratory, U.S. Geological Survey, Biological Resources Division, 1700 Leetown Road, Kearneysville, West Virginia 25430, USA
2The Conservation Fund, Freshwater Institute, PO Box 1746, Shepherdstown, West Virginia 25443, USA

ABSTRACT: Growth of Renibacterium salmoninarum was compared in 14 different broth media; 13 serum-free, and 1 that contained newborn calf serum, KDM2+M. Supplementation with 1% v/v R. salmoninarum MCO4M metabolite was evaluated for 6 of the media that do not utilize it as part of their ingredients. Viable cells were enumerated on Days 10, 20, and 30 post inoculation to evaluate performance. The experiment was repeated 3 times using high, low, and medium (trials 1 to 3, respectively) cell concentrations as inoculum. In general there was no optimal medium and all performed well. The choice of which to employ depends on the ease of preparation and presence of certain ingredients that might affect subsequent assays. In trials 2 and 3, the pH was estimated using test papers at the same time as cells were counted. Maximum pH increase occurred with KDM2+M and those media containing charcoal. For most media, a simple pH determination could be used as a means to check that growth has occurred in a culture, particularly if charcoal was added directly to the media and a visual inspection could not be made to detect growth.

KEY WORDS: Renibacterium salmoninarum · Bacterial Kidney Disease · Broth media · Serum free

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