Background:
The surface of polymeric tracheotomy tubes is a favourable environment for biofilm formation and therefore represents a potential risk factor for the development of pneumonia after tracheotomy. The aim of this in-vitro study was to develop octenidine dihydrochloride (OCT) coated polymer tracheotomy tubes and investigate any effects on Staphylococcus aureus and Pseudomonas aeruginosa colonization. Additionally the resistance of the OCT coating was tested using reprocessing procedures like brushing, rinsing and disinfection with glutaraldehyde.
Results:
Contamination with S. aureus: Before any reprocessing, OCT coated tracheotomy tubes were colonized with 103 cfu/ml and uncoated tracheotomy tubes with 105 10 cfu/ml (P=0.045). After reprocessing, no differences in bacterial concentration between modified and conventional tubes were observed. Contamination with P. aeruginosa: Before reprocessing, OCT coated tubes were colonized with 106 cfu/ml and uncoated tubes with 107 cfu/ml (P=0.006). After reprocessing, no significant differences were observed.
Conclusion:
OCT coating initially inhibits S. aureus and P. aeruginosa colonisation on tracheotomy tubes. This effect, however, vanishes quickly after reprocessing of the tubes due to poor adhesive properties of the antimicrobial compound. Despite the known antimicrobial effect of OCT, its use for antimicrobial coating of tracheotomy tubes is limited unless methods are developed to allow sustained attachment to the tube.

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Background:
The development of a new cold-active beta-D-galactosidase and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production.
Results:
In this article, we present a new beta-D-galactosidase as a candidate to be applied in the above mentioned biotechnological processes. The gene encoding this beta-D-galactosidase has been isolated from the genomic DNA library of Antarctic bacterium Arthrobacter sp. 32c, sequenced, cloned, expressed in Escherichia coli and Pichia pastoris, purified and characterized. 27 mg of beta-D-galactosidase was purified from 1 L of culture with the use of an intracellular E. coli expression system. The protein was also produced extracellularly by P. pastoris in high amounts giving approximately 137 mg and 97 mg of purified enzyme from 1 L of P. pastoris culture for the AOX1 and a constitutive system, respectively. The enzyme was purified to electrophoretic homogeneity by using either one step- or a fast two step- procedure including protein precipitation and affinity chromatography. The enzyme was found to be active as a homotrimeric protein consisting of 695 amino acid residues in each monomer. Although, the maximum activity of the enzyme was determined at pH 6.5 and 50degreesC, 60% of the maximum activity of the enzyme was determined at 25degreesC and 15% of the maximum activity was detected at 0degreesC.
Conclusions:
The properties of Arthrobacter sp. 32c beta-D-galactosidase suggest that this enzyme could be useful for low-cost, industrial conversion of lactose into galactose and glucose in milk products and could be an interesting alternative for the production of ethanol from lactose-based feedstock.

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Background:
Acquiring a highly stable photonic plasmid in transformed Salmonella typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella typhimurium (S. typh-lux) using three different plasmids and characterize their respective photonic properties.
Results:
In presence of ampicillin (AMP), S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 plasmids exhibited 100% photon-emitting colonies over a 10-d study period. Photon emitters of S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 without AMP selection decreased over time (P < 0.05), representing only 11 +/- 1%, 35 +/- 1% and 43 +/- 1%, respectively, of original photon emitting properties of the bacterial population by d 10. Photonic emissions were positively correlated with bacterial concentration (P < 0.05) for pAK1-lux, pCGLS-1 and pXEN-1 (r = 0.96, 0.98 and 0.82, respectively). When stratified by high, medium and low density bacteria concentrations, photonic emissions for high density populations containing pAK1-lux, pCGLS-1 and pXEN-1 resulted in differences of photonic emissions across a range of bacterial concentrations (1×10^7 to 1×10^9 CFU, P < 0.05) with positive correlations (P < 0.05) of (r = 0.72, 0.46 and 0.72, respectively). The correlation of photonic emissions with bacterial concentrations for samples with medium and low density bacteria (pAK1-lux, pCGLS-1, and pXEN-1 plasmids) imaged in tubes were also positively correlated (medium; r=0.69, 0.49, 0.46, low; r=0.90, 0.71, 0.68, respectively; P > 0.05); although photonic emissions across a range of bacterial concentrations were not different (1×10^4 to 1×10^6 CFU, P > 0.05). For very low density bacterial concentrations imaged in 96 well plates photonic emissions were positively correlated with bacterial concentration (P < 0.05) for pAK1-lux, pCGLS-1, and pXEN-1 plasmids (r=0.99, 0.99, and 0.96, respectively), and photonic emissions across a range of bacterial concentrations (1×10^3 to 1×10^5 CFU) low to high were different in the 96-well plate format (P < 0.05).
Conclusion:
These data characterize photon stability properties for S. typh-lux transformed with three different photon generating plasmids that may facilitate real-time Salmonella tracking using in vivo or in situ biophotonic paradigms. [USDA-ARS Biophotonics Initiative # 58-6402-3-0120].

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Background:
Enteropathogenic Escherichia coli (EPEC) produce attaching/effacing (A/E) lesions on eukaryotic cells mediated by the outer membrane adhesin intimin. EPEC are sub-grouped into typical (tEPEC) and atypical (aEPEC). We have recently demonstrated that aEPEC strain 1551-2 (serotype O non-typable, non-motile) invades HeLa cells by a process dependent on the expression of intimin sub-type omicron. In this study, we evaluated whether aEPEC strains expressing other intimin sub-types are also invasive using the quantitative gentamicin protection assay. We also evaluated whether aEPEC invade differentiated intestinal T84 cells.
Results:
Five of six strains invaded HeLa and T84 cells in a range of 13.3%-20.9% and 5.8%-17.8%, respectively, of the total cell-associated bacteria. The strains studied were significantly more invasive than prototype tEPEC strain E2348/69 (1.4% and 0.5% in HeLa and T84 cells, respectively). Invasiveness was confirmed by transmission electron microscopy. We also showed that invasion of HeLa cells by aEPEC 1551-2 depended on actin filaments, but not on microtubules. In addition, disruption of tight junctions enhanced its invasion efficiency in T84 cells, suggesting preferential invasion via a non-differentiated surface.
Conclusion:
Some aEPEC strains may invade intestinal cells in vitro with varying efficiencies and independently of the intimin sub-type.

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