Multifractal Analysis to Determine the Effect of Surface Topography on the Distribution, Density, Dispersion and Clustering of Differently Organised Coccal-Shaped Bacteria.

The topographic features of surfaces are known to affect bacterial retention on a surface, but the precise mechanisms of this phenomenon are little understood. Four coccal-shaped bacteria, Staphylococcus sciuri, Streptococcus pyogenes, Micrococcus luteus, and Staphylococcus aureus, that organise in different cellular groupings (grape-like clusters, tetrad-arranging clusters, short chains, and diploid arrangement, respectively) were used. These differently grouped cells were used to determine how surface topography affected their distribution, density, dispersion, and clustering when retained on titanium surfaces with defined topographies. Titanium-coated surfaces that were smooth and had grooved features of 1.02 µm-wide, 0.21 µm-deep grooves, and 0.59 µm-wide, 0.17 µm-deep grooves were used. The average contact angle of the surfaces was 91°. All bacterial species were overall of a hydrophobic nature, although M. luteus was the least hydrophobic. It was demonstrated that the 1.02 µm-wide featured surface most affected Strep. pyogenes and S. sciuri, and hence the surfaces with the larger surface features most affected the cells with smaller dimensions. The 0.59 µm featured surface only affected the density of the bacteria, and it may be suggested that the surfaces with the smaller features reduced bacterial retention. These results demonstrate that the size of the topographical surface features affect the distribution, density, dispersion, and clustering of bacteria across surfaces, and this is related to the cellular organisation of the bacterial species. The results from this work inform how surface topographical and bacterial properties affect the distribution, density, dispersion, and clustering of bacterial retention.

Factors Involved in the onset of infection following bacterially contaminated platelet transfusions.

Transfusion of platelet concentrates (PCs) is associated with several adverse patient reactions, the most common of which are febrile non-hemolytic transfusion reactions (FNHTRs) and transfusion-associated bacterial-infection/transfusion-associated sepsis (T-ABI/TA-S). Diagnosis of T-ABI/T-AS requires a positive blood culture (BC) result from the transfusion recipient and also a positive identification of bacterial contamination within a test aliquot of the transfused PC. In a significant number of cases, clinical symptoms post-transfusion are reported by the clinician, yet the BCs from the patient and/or PC are negative. The topic of 'missed bacterial detection' has therefore been the focus of several primary research studies and review articles, suggesting that biofilm formation in the blood bag and the presence of viable but non-culturable (VBNC) pathogens are the major causes of this missed detection. However, platelets are emerging as key players in early host responses to infection and as such, the aforementioned biofilm formation could elicit 'platelet priming', which could lead to significant immunological reactions in the host, in the absence of planktonic bacteria in the host bloodstream. This review reflects on what is known about missed detection and relates this to the emerging understanding of the effect of bacterial contamination on the platelets themselves and the significant role played by platelets in exacerbation of an immune response to infection within the transfusion setting.

Molybdenum Disulfide Surfaces to Reduce Staphylococcus aureus and Pseudomonas aeruginosa Biofilm Formation.

The reduction of bacteria and biofilm formation is important when designing surfaces for use in industry. Molybdenum disulfide surfaces (MoS2SUR) were produced using MoS2 particle (MoS2PAR) sizes of 90 nm, 2 µm, and 6 µm containing MoS2PAR concentrations of 5%, 10%, 15%, and 20%. These were tested to determine the efficacy of the MoS2SUR to impede bacterial retention and biofilm formation of two different types of bacteria, Staphylococcus aureus and Pseudomonas aeruginosa. The MoS2SUR were characterized using Fourier transform infrared spectroscopy, ion-coupled plasma atomic emission spectroscopy, scanning electron microscopy, optical profilometry, and water contact angles. The MoS2SUR made with the smaller 90 nm MoS2PAR sizes demonstrated smaller topographical-shaped features. As the size of the incorporated MoS2PAR increased, the MoS2SUR demonstrated wider surface features, and they were less wettable. The increase in MoS2PAR concentration within the MoS2SUR groups did not affect the surface topography but did increase wettability. However, the increase in MoS2PAR size increased both the surface topography and wettability. The MoS2SUR with the smaller topographical-shaped features influenced the retention of the S. aureus bacteria. Increased MoS2SUR topography and wettability resulted in the greatest reduction in bacterial retention, and the bacteria became more heterogeneously dispersed and less clustered across the surfaces. The surfaces that exhibited decreased bacterial retention (largest particle sizes, largest features, greatest roughness, and most wettable) resulted in decreased biofilm formation. Cytotoxicity testing of the surface using cell viability demonstrated that the MoS2SUR were not toxic against HK-2 cells at MoS2PAR sizes of 90 nm and 2 µm. This work demonstrated that individual surface variables (MoS2SUR topographic shape and roughness, MoS2PAR size, and concentration) decreased bacterial loading on the surfaces, which then decreased biofilm formation. By optimizing MoS2SUR properties, it was possible to impede bacterial retention and subsequent biofilm formation.

The effects of blood conditioning films on the antimicrobial and retention properties of zirconium-nitride silver surfaces.

External bone fixation devices provide support and rehabilitation for severely damaged/broken bones, however, this invasive procedure is prone to infection. Zirconium nitride/silver (Ti-ZrN/Ag) coatings were characterised for surface topography, chemical composition, physicochemistry and antimicrobial efficacy (against Staphylococcus aureus and Staphylococcus epidermidis), in the presence of a blood conditioning film. The conditioning film altered the width of the microtopography of the surfaces however, the depth of the features remained relatively constant. The conditioning film also altered the coatings from hydrophobic to hydrophilic/partially hydrophilic surfaces. Following the MATH assay, the presence of a conditioning film reduced affinity towards the hydrocarbons for both microorganisms. The addition of a blood conditioning film reduced the antimicrobial efficacy of the Ti-ZrN/Ag coatings but also reduced the number of retained bacteria. This study suggests that the presence of a pre-defined blood conditioning film may result in surfaces with anti-adhesive properties, potentially leading to a reduction in bacterial retention. This, combined with the antimicrobial efficacy of the coatings, could reduce the risk of infection on biomaterial surfaces.

Antimicrobial synergy of cationic grafted poly(para-phenylene ethynylene) and poly(para-phenylene vinylene) compounds with UV or metal ions against Enterococcus faecium.

The rise in multidrug resistant bacteria is an area of growing concern and it is essential to identify new biocidal agents. Cationic grafted compounds were investigated for their antimicrobial properties using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) tests. Synergy testing was carried out using the compounds in the presence of ultraviolet (UV). Fractional inhibitory concentration (FIC) and fractional bactericidal concentration (FBC) tests were carried out using the cationic molecules in conjunction with metal ion solutions of gold, silver, palladium, platinum, rhodium, titanium, tin, vanadium and molybdenum. Individually, the cationic compounds containing quaternary amines, polyphenylene vinylene (PPV) with long polyacrylate grafts (PPV-g-PMETAC (HMw)), polyphenylene ethylene (PPE) with long polyacrylate grafts (PPE-g-PMETAC (HMw)), polyphenylene vinylene (PPV) with short polyacrylate grafts (PPV-g-PMETAC (LMw)) and polyphenylene ethylene (PPE) with short polyacrylate grafts (PPE-g-PMETAC (LMw)) were effective against Enterococcus faecium. The most successful compound under UV was PPV-g-PMETAC (HMw). Following the FICs, palladium and rhodium ion solutions caused a synergistic reaction with all four tested compounds. The presence of conjugated bonds in the cationic molecules increased its antimicrobial activity. These results suggest that the chemical backbone of the compounds, alongside the chain lengths and chain attachment affect the antimicrobial efficacy of a compound. These factors should be taken into consideration when formulating new biocidal combinations.

Surface modification of platelet concentrate bags to reduce biofilm formation and transfusion sepsis.

Bacterial contamination of blood products poses a major risk in transfusion medicine, including transfusions involving platelet products. Although testing systems are in place for routine screening of platelet units, the formation of bacterial biofilms in such units may decrease the likelihood that bacteria will be detected. This work determined the surface properties of p-PVC platelet concentrate bags and investigated how these characteristics influenced biofilm formation. Serratia marcescens and Staphylococcus epidermidis, two species commonly implicated in platelet contamination, were used to study biofilm growth. The platelet concentrate bags were physically flattened to determine if reducing the surface roughness altered biofilm formation. The results demonstrated that the flattening process of the platelet bags affected the chemistry of the surface and reduced the surface hydrophobicity. Flattening of the surfaces resulted in a reduction in biofilm formation for both species after 5 days, with S. marcescens demonstrating a greater reduction. However, there was no significant difference between the smooth and flat surfaces following 7 days' incubation for S. marcescens and no significant differences between any of the surfaces following 7 days' incubation for S. epidermidis. The results suggest that flattening the p-PVC surfaces may limit potential biofilm formation for the current duration of platelet storage time of 5 days. It is hoped that this work will enhance the understanding of how surface properties influence the development of microbial biofilms in platelet concentrate bags in order to devise a solution to discourage biofilm formation.