Characterization of biofilm-forming abilities of antibiotic-resistant Salmonella typhimurium DT104 on hydrophobic abiotic surfaces.
ABSTRACT Salmonella typhimurium DT104 strain has emerged as a global human and veterinary public health concern because of its antibiotic resistance and extensive host range. Although it is thought to be more virulent, to date, factors relevant to its virulence have not been fully elucidated. Thus, understanding how this strain forms biofilms on hydrophobic surfaces will add to current knowledge on its possible virulence mechanism.
Biofilm-forming abilities of clinical isolates of S. typhimurium DT104 from human and animal sources on hydrophobic inanimate surfaces were assessed by absorbance at 600 nm of crystal violet-bound cells recovered from 96-well tissue culture plates after growth in a nutrient-rich growth medium and various adjusted media; and scanning electron microscopy based on standard procedures.
In the nutrient-rich growth medium, Luria-Bertani (LB), biofilms were formed in small quantities, preferentially on polystyrene (p<0.05), and followed different time courses. Significantly lower amounts of biofilms were formed on polystyrene when a nutrient-deficient growth medium (adherence test medium) was used. Inclusion of D-(+)-mannose in LB at a concentration of 100 mM significantly (p<0.05) inhibited biofilm formation on polystyrene. D-(+)-glucose relatively enhanced biofilm formation but D-(-)-mannitol only insignificantly influenced the process. The action of mannose on polyvinly chloride (PVC) was insignificant, suggesting that its action may be surface-dependent. Additionally, glucose significantly reduced biofilm growths of 2 of the isolates and only that of the PVC-loving strain T980021 on polystyrene and PVC, respectively. At the concentration tested, unlike xylose, both D-mannose and D-glucose significantly (p<0.05) inhibited bacterial growth, providing a possible mechanism for their inhibitory action on biofilm formation by S. typhimurium. While stress of starvation resulted in significant reduction in biofilm formation on polystyrene in all but the PVC-loving strain T980021, high osmolarity had little effect on the quantity of biofilm formed on polystyrene. The extent of primary attachment to polystyrene as well as their capacity to form biofilm did not correlate with their cell surface hydrophobicity and exopolysaccharide production.
D-(+)-mannose inhibits biofilm formation by S. typhimurium DT104 on polystyrene but not on PVC. There was also a general lack of correlation between the ability of S. typhimurium DT104 to form biofilm and its physicochemical surface characteristics.
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ABSTRACT: This study was designed to evaluate gene expression patterns of the planktonic and biofilm cells of Staphylococcus aureus and Salmonella Typhimurium in trypticase soy broth adjusted to pH 5.5 and pH 7.3. The planktonic and biofilm cells of multiple antibiotic-resistant S. aureus (S. aureus(R) ) and S. Typhimurium (S. Typhimurium(R) ) were more resistant to β-lactams than those of antibiotic-susceptible S. aureus (S. aureus(S) ) and S. Typhimurium (S. Typhimurium(S) ) at pH 5.5 and pH 7.3. The relative gene expression levels of norB, norC, and mdeA genes were increased by 7.0-, 4.7-, and 4.6-fold, respectively, in the biofilm cells of S. aureus(S) grown at pH 7.3, while norB, norC, mdeA, sec, seg, sei, sel, sem, sen, and seo genes were stable in the biofilm cells of S. aureus(R) . This study provides useful information for understanding gene expression patterns in the planktonic and biofilm cells of antibiotic-resistance pathogens exposed to acidic stress.FEMS Microbiology Letters 10/2011; 325(2):180-8. · 2.72 Impact Factor
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ABSTRACT: Bacterial biofilms are crucial to the pathogenesis of many important infections and are difficult to eradicate. Streptococcus suis is an important pathogen of pigs, and here the biofilm-forming ability of 32 strains of this species was determined. Significant biofilms were completely formed by 10 of the strains after 60 h of incubation, with exopolysaccharide production in the biofilm significantly higher than that in the corresponding planktonic cultures. S. suis strain SS2-4 formed a dense biofilm, as revealed by scanning electron microscopy, and in this state exhibited increased resistance to a number of antibiotics (ampicillin, amoxicillin, ciprofloxacin, kanamycin, and rifampin) compared to that of planktonic cultures. A bacteriophage lysin, designated LySMP, was used to attack biofilms alone and in combination with antibiotics and bacteriophage. The results demonstrated that the biofilms formed by S. suis, especially strains SS2-4 and SS2-H, could be dispersed by LySMP and with >80% removal compared to a biofilm reduction by treatment with either antibiotics or bacteriophage alone of less than 20%; in addition to disruption of the biofilm structure, the S. suis cells themselves were inactivated by LySMP. The efficacy of LySMP was not dose dependent, and in combination with antibiotics, it acted synergistically to maximize dispersal of the S. suis biofilm and inactivate the released cells. These data suggest that bacteriophage lysin could form part of an effective strategy to treat S. suis infections and represents a new class of antibiofilm agents.Applied and Environmental Microbiology 12/2011; 77(23):8272-9. · 3.95 Impact Factor
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ABSTRACT: Scorpine, a small cationic peptide from the venom of Pandinus imperator, which has been shown to have anti-bacterial and anti-plasmodial activities, has potential important applications in the pharmaceutical industries. However, the isolation of scorpine from natural sources is inefficient and time-consuming. Here, we first report the expression and purification of recombinant scorpine in Escherichia coli, using small ubiquitin-related modifier (SUMO) fusion partner. The fusion protein was expressed in soluble form in E. coli, and expression was verified by SDS-PAGE and western blotting analysis. The fusion protein was purified to 90% purity by nickel-nitrilotriacetic acid (Ni2+-NTA) resin chromatography. After the SUMO-scorpine fusion protein was cleaved by the SUMO protease, the cleaved sample was reapplied to a Ni2+-NTA column. Tricine/SDS-PAGE gel results indicated that Scorpine had been purified successfully to more than 95% purity. The recombinantly expressed Scorpine showed anti-bacterial activity against two standard bacteria including Staphylococcus aureus ATCC 29213 and Acinetobacter baumannii ATCC 19606, and clinically isolated bacteria including S. aureus S, S. aureus R, A. baumannii S, and A. baumannii R. It also produced 100% reduction in Plasmodium falciparum parasitemia in vitro. Thus, the expression strategy presented in this study allowed convenient high yield and easy purification of recombinant Scorpine for pharmaceutical applications in the future.PLoS ONE 07/2014; 9(7):e103456. · 3.53 Impact Factor