Lipase, protease, and biofilm as the major virulence factors in staphylococci isolated from acne lesions.

Staphylococci involve infections in association with a number of bacterial virulence factors. Extracellular enzymes play an important role in staphylococcal pathogenesis. In addition, biofilm is known to be associated with their virulence. In this study, 149 staphylococcal isolates from acne lesions were investigated for their virulence factors including lipase, protease, and biofilm formation. Coagulase-negative staphylococci were demonstrated to present lipase and protease activities more often than coagulase-positive staphylococci. A microtiter plate method (quantitative method) and a Congo red agar method (qualitative method) were comparatively employed to assess biofilm formation. In addition, biofilm forming ability was commonly detected in a coagulase-negative group (97.7%, microtiter plate method and 84.7%, Congo red agar method) more frequently than in coagulase-positive organisms (68.8%, microtiter plate method and 62.5%, Congo red agar method). This study clearly confirms an important role for biofilm in coagulasenegative staphylococci which is of serious concern as a considerable infectious agent in patients with acnes and implanted medical devices. The Congo red agar method proved to be an easy method to quickly detect biofilm producers. Sensitivity of the Congo red agar method was 85.54% and 68.18% and accuracy was 84.7% and 62.5% in coagulase-negative and coagulase-positive staphylococci, respectively, while specificity was 50% in both groups. The results clearly demonstrated that a higher percentage of coagulasenegative staphylococci isolated from acne lesions exhibited lipase and protease activities, as well as biofilm formation, than coagulase-positive staphylococci.

Rhodomyrtus tomentosa (Aiton) Hassk. ethanol extract and rhodomyrtone: a potential strategy for the treatment of biofilm-forming staphylococci.

The anti-staphylococcal activity of an ethanol extract of Rhodomyrtus tomentosa and its pure compound, rhodomyrtone, as well as their effects on staphylococcal biofilm formation and biofilm-grown cells were assessed. MIC and minimal bactericidal concentration values of the ethanol extract and rhodomyrtone against planktonic cultures and biofilms of five clinical strains each of Staphylococcus aureus and Staphylococcus epidermidis, and American Type Culture Collection (ATCC) strains of both species, were 32-512 and 0.25-2 µg ml(-1), respectively. Results from time-kill studies indicated that rhodomyrtone at a concentration of 4× MIC could reduce the number of Staphylococcus aureus ATCC 25923 and Staphylococcus epidermidis ATCC 35984 cells by 99.9% within 3 and 13 h, respectively. The ability of rhodomyrtone and the ethanol extract to prevent biofilm formation and kill mature biofilms was assessed: both demonstrated better activity than vancomycin at inhibiting staphylococcal biofilm formation. In addition, the viability of 24 h and 5-day staphylococcal biofilm-grown cells decreased after treatment with the ethanol extract and rhodomyrtone. The ability to reduce biofilm formation and kill mature biofilms occurred in a dose-dependent manner. Scanning electron microscopy clearly confirmed that treatment with rhodomyrtone at 16× MIC could reduce 24 h biofilm formation and the numbers of staphylococci, whilst at 64× MIC this compound destroyed the organisms in the 5-day established biofilm. These results suggest that rhodomyrtone has the potential for further drug development for the treatment of biofilm-forming staphylococcal infections.

Characterization of the antiyeast compound and probiotic properties of a starter lactobacillus plantarum DW3 for possible use in fermented plant beverages

Lactobacillus plantarum DW3 produced antifungal compounds that inhibited the growth of Rhodotorula mucilaginosa DKA, contaminating yeast in fermented plant beverages (FPBs) and various potential human pathogens. Phenyllactic acid (PLA) identified by gas chromatography- mass spectrometry (GC-MS) was produced at 31 mg/L PLA in MRS medium and 5 mg/ml inhibited growth of the target yeast in vitro by 90 percent. Other inhibitors were also present but not specifically identified. Results of in vitro tests showed that DW3 also had probiotic properties as it survived various human biological barriers resistance to pH 3, bile salts, growth without vitamin B12 and the presence and absence of oxygen. Its inhibitory effect against food borne pathogenic bacteria and spoilage organisms was higher than that found for a commercial strain Lactobacillus casei R. An acute oral toxicity test on ICR mice at a high single dose of either 10(9) and 10(12) cells per mouse for 14 days showed that DW3 had no adverse effect on the general health status and there was no evidence of bacteremia. Mice fed DW3 had a reduced weight gain compared to the control. No significant difference (p > 0.05) was found for the spleen weight index (SWI) among the treatment and control groups whereas there was a significant difference (p < 0.05) for the liver weight ratio (LWR) in a group fed with 10(12) cells per mouse when compared with the control group.