BackgroundStaphylococcus aureus is a major human pathogen causing significant morbidity and mortality. The S. aureus colonies in osteomyelitis, in patients with cystic fibrosis and patients with endoprosthesis rejection frequently have an atypical morphology, i.e. staphylococcal small-colony variants, which form a naturally occurring subpopulation of clinically important staphylococci. Identification of these small colony variants is difficult, because of the loss of typical phenotypic characteristics of these variants.We wanted to improve and simplify the diagnosis of staphylococcal infection using a diagnostic preparation, consisting of 5 μg batumin paper disks. Batumin possesses a unique selective activity against all studied Staphylococcus spp., whereas all other species tested thus far are batumin resistant. We assessed the efficacy of the batumin diagnostic preparation to identify staphylococcal small colony variants, isolated from osteomyelitis patients.FindingsWith the batumin diagnostic preparation, all 30 tested staphylococcal small-colony variants had a growth inhibition zone around the disk of minimum 25 mm, accordant with the inhibition zones of the parent strains, isolated from the same patients.ConclusionsThe batumin diagnostic preparation correctly identified the small-colony variants of S. aureus, S. haemolyticus and S. epidermidis as belonging to the genus Staphylococcus, which differ profoundly from parental strains and are difficult to identify with standard methods. Identification of staphylococcal small-colony variants with the batumin diagnostic preparation is technically simple and can facilitate practical laboratory work.
The antibiotic batumin, produced by Pseudomonas batumici, has been shown to be highly active against 123 type and reference strains and clinical isolates of 30 Staphylococcus species (including MRSA and small colony variants-(SSCVs) of S. aureus, S. epidermidis and S. haemolyticus). Batumin activity against these bacteria did not depend on the species, origin or resistance to other antibiotics and its MIC was 0.0625 -0.5 µg/ml. Batumin influence on biofilm formation was studied in clinical isolates of S. aureus, S. epidermidis and S. intermedius. Addition of batumin at a concentration of half of the MIC in the broth, i.e. 0.125 µg/ml, decreased the biofilm of 16 out of 20 S. aureus strains to varying degrees. Batumin was more effective against Staphylococcus strains with strong biofilm formation. Using atomic-force microscopy, it could be shown that batumin reduced the number of S. aureus ATCC 25923 adherent cells more than fourfold. The adherent cells of staphylococci were visualized as monolayers of separate islets. A detailed study of the surface of bacterial cells treated with batumin allowed to establish significant reduction of their roughness values. Observed values were typical for planktonic S. aureus cells. The obtained data explain one of the mechanisms of the antimicrobial activity of batumin, which is based оn preventing the formation of S. aureus biofilm. As such, batumin could be considered as an agent offering opportunities
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