Biology Faculty Articles
Title
Assays optimized for detection and quantification of antibacterial activity in shark cell lysates under high salt conditions
Document Type
Article
Publication Date
1-1-2013
Publication Title
Fish and Shellfish Immunology
Keywords
Shark, Antibacterial assays, Leukocyte lysates
ISSN
1050-4648
Volume
34
Issue/No.
5
First Page
1223
Last Page
1227
Abstract
To assess non-cellular innate immune mechanisms that play a role in the antimicrobial defense of an organism several assay systems have been devised to screen for such factors. Most assays, however, have been developed to measure activity against clinical isolates of medical importance. There is scant information on methods optimal for assaying material from sharks and other marine fish for antimicrobial activity particularly against salt tolerant organisms that are likely to be encountered in the marine environment. We have modified and optimized agar diffusion and broth dilution assays for detection and quantification of antibacterial activity of shark leukocyte lysates. By replacing marine agar, typically used for marine organisms, with artificial sea water complete medium (SCM) enriched with tryptone and yeast extract has resulted in an improved inhibition zone assay that uses Planococcus citreus, a salt-tolerant organism as the target organism. Antibacterial activity is correlated to the size of zone of no bacterial growth around wells containing bioactive test sample. An alternative broth based microdilution growth assay uses the 96 well format and the antibacterial effect of the sample on growth of P. citreus, the target organism, is measured spectrophotometrically as percent inhibition of bacterial growth when compared to the growth of P. citreus grown in medium alone that represents 100% growth. The assay can also be used to titrate antibacterial activity and express the level of growth inhibition as a titer.
NSUWorks Citation
Smith, Sylvia L. and Nichole Vaughan. 2013. "Assays optimized for detection and quantification of antibacterial activity in shark cell lysates under high salt conditions." Fish and Shellfish Immunology 34, (5): 1223-1227. https://nsuworks.nova.edu/cnso_bio_facarticles/51
