ABSTRACT
A polyphasic taxonomic study using morphological, biochemical, chemotaxonomic and molecular methods was performed on three strains of a Gram-stain positive, non-sporeforming, motile aerobic rod-shaped bacterium resistant to tylosin and tetracycline isolated from a swine-manure storage pit. On the basis of 16S rRNA gene sequence analyses, it was confirmed that these isolates are highly related to each other and form a hitherto unknown lineage within the Planococcaceae. In particular, pairwise analysis of the 16S rRNA gene sequence demonstrated that the novel organism is closely related to members of the genus Sporosarcina (92.8-94.5 %), Pyschrobacillus (93.5-93.9 %) and Paenisporosarcina (93.3-94.5 %). The predominant fatty acids were found to consist of iso-C15:0 and iso-C17:1 ω10c and the G+C mol% was determined to be 41.8. Based on biochemical, chemotaxonomic, and phylogenetic evidence, it is proposed that these novel strains be classified as a novel genus and species, Savagea faecisuis gen nov., sp. nov. The type strain is Con12(T) (=CCUG 63563(T) = NRRL B-59945(T) = NBRC 109956(T)).
Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Environmental Microbiology , Planococcaceae/classification , Planococcaceae/isolation & purification , Tetracycline/pharmacology , Tylosin/pharmacology , Aerobiosis , Animals , Base Composition , Cluster Analysis , Cytosol/chemistry , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Fatty Acids/analysis , Locomotion , Manure , Molecular Sequence Data , Phylogeny , Planococcaceae/drug effects , Planococcaceae/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , SwineABSTRACT
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.
