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Screening bacterial metabolites for inhibitory effects against Batrachochytrium dendrobatidis using a spectrophotometric assay

Bell, S. C. and Alford, R. A. and Garland, S. and Padilla, G. and Thomas, A. D. (2013) Screening bacterial metabolites for inhibitory effects against Batrachochytrium dendrobatidis using a spectrophotometric assay. Diseases of Aquatic Organisms, 103 (1). 77-+. ISSN 0177-5103

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Article Link(s): http://dx.doi.org/10.3354/dao02560

Abstract

Certain bacteria present on frog skin can prevent infection by the pathogenic fungus Batrachochytrium dendrobatidis (Bd), conferring disease resistance. Previous studies have used agar-based in vitro challenge assays to screen bacteria for Bd-inhibitory activity and to identify candidates for bacterial supplementation trials. However, agar-based assays can be difficult to set up and to replicate reliably. To overcome these difficulties, we developed a semi-quantitative spectrophotometric challenge assay technique. Cell-free supernatants were prepared from filtered bacterial cultures and added to 96-well plates in replicated wells containing Bd zoospores suspended in tryptone-gelatin hydrolysate-lactose (TGhL) broth medium. Plates were then read daily on a spectrophotometer until positive controls reached maximum growth in order to determine growth curves for Bd. We tested the technique by screening skin bacteria from the Australian green-eyed tree frog Litoria serrata. Of bacteria tested, 31% showed some degree of Bd inhibition, while some may have promoted Bd growth, a previously unknown effect. Our cell-free supernatant challenge assay technique is an effective in vitro method for screening bacterial isolates for strong Bd-inhibitory activity. It contributes to the expanding field of bioaugmentation research, which could play a significant role in mitigating the effects of chytridiomycosis on amphibians around the world.

Item Type:Article
Business groups:Biosecurity Queensland
Additional Information:Bell, Sara C. Alford, Ross A. Garland, Stephen Padilla, Gabriel Thomas, Annette D.
Subjects:Aquaculture and Fisheries > Aquaculture > Frog culture
Veterinary medicine > Communicable diseases of animals (General)
Deposited On:25 Oct 2013 06:45
Last Modified:25 Oct 2013 06:46

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