Reduction In Bacterial Numbers Research Articles

Strong antibacterial dental resin composites containing cellulose nanocrystal/zinc oxide nanohybrids

ObjectiveThe aim is to explore the reinforcing and antibacterial effect of cellulose nanocrystal/zinc oxide (CNC/ZnO) nanohybrids on dental resin composites (DRCs). MethodsCNC/ZnO nanohybrids were prepared through precipitating Zn2+ on the surface of CNC and then introduced into the DRCs. The mechanical properties of DRCs including compressive strength, flexural modulus, flexural strength, and Vickers microhardness were characterized. The antibacterial activity of DRCs to Streptococcus mutans was determined and the morphology of Streptococcus mutans on the surface of DRCs after incubation was observed. The morphology of fractured surface after flexural test and Zn content in DRCs were analyzed. ResultsCompared with DRCs without CNC/ZnO nanohybrids, DRCs containing 2 wt.% CNC/ZnO nanohybrids possess higher compressive strength and flexural modulus and there is no significantly statistical difference (P＞0.05) on the flexural strength and Vickers microhardness. The excess use of CNC/ZnO nanohybrids decreases the mechanical properties of DRCs except flexural modulus. DRCs containing CNC/ZnO nanohybrids show excellent antibacterial properties and a 78% reduction in bacterial number is obtained when 2% CNC/ZnO nanohybrids are added. ConclusionThe small amounts of CNC/ZnO nanohybrids have a positive influence on the mechanical and antibacterial properties of DRCs. SignificanceThe prepared DRCs are promising to address bulk fracture and secondary caries.

The essential role of complement in antibody-mediated resistance to Salmonella.

SummaryVaccines can serve as essential tools to prevent bacterial diseases via the induction of long‐lasting IgG responses. The efficacy of such vaccines depends on the effector mechanisms triggered by IgG. The complement system and Fc‐gamma receptors (FcγRs) can potentially play a crucial role in IgG‐mediated immunity against bacterial diseases. However, their relative importance in vivo is unclear, and has been the object of controversy and debate. In this brief study, we have used gene‐targeted mice lacking either Fcγ RI, II, II and IV or the C3 complement component as well as a novel mouse strain lacking both C3 and FcγRs to conclusively show the essential role of complement in antibody‐mediated host resistance to Salmonella enterica systemic infection. By comparing the effect of IgG2a antibodies against Salmonella O‐antigen in gene‐targeted mice, we demonstrate that the complement system is essential for the IgG‐mediated reduction of bacterial numbers in the tissues.

Detection of the unknown components of the oral microflora of teeth with periapical radiolucencies in a Turkish population using next-generation sequencing techniques.

To detect the unknown components of the oral microbiome and the effects of root canal treatment in a Turkish population and to evaluate the changes in microbial diversity in the root canals before and after treatment. Single-rooted central and lateral maxillary incisors with one canal were chosen from 20 patients. Baseline samples of intact intracanal microbiota were collected from 20 root canals of permanent teeth with necrotic pulps using sterile paper points. After root canal preparation, the root canals were filled with a calcium hydroxide paste for 7days. Calcium hydroxide was removed from root canal with 2.5% sodium hypochlorite and 17% EDTA using passive ultrasonic irrigation (PUI). A second bacteriologic samples were taken with sterile paper points prior to root filling. The samples were processes with DNase-I treatment using next-generation sequencing (NGS). Reduction in bacterial numbers during root canal treatment was evaluated using real-time quantitative PCR (qPCR). All statistical analyses were conducted using the MINITAB 17 software (Minitab Ltd. Co., Coventry, UK). A one-sample t-test was used to analyse the data. Statistical significance was accepted at P<0.05. Relative abundances of Mycoplasma sp., Paludibacter sp., Tannerella sp., Prevotella spp. and an uncultured species from the order Bacteroidales decreased with root canal preparation and medication (98.7%, 99.8%, 98.8%, 97.7% and 99.3%, respectively), whilst the relative abundances of Methylobacterium sp., Corynebacterium sp. and Streptococcus infantis increased (93.1%, 94.8% and 99.4%, respectively). Considerable numbers of Streptophyta species were detected before and after treatment. The ratio of Agrobacterium sp. in the treated teeth community and the ratio of order Streptophyta in the infected canals had negative correlations with the success of bacterial elimination. The combination of NGS and qPCR techniques resulted in detection of previously unknown components of the oral microbiome and the effects of root canal treatment on their relative abundance in a Turkish population.

Clostridium sordellii outer spore proteins maintain spore structural integrity and promote bacterial clearance from the gastrointestinal tract

Bacterial spores play an important role in disease initiation, transmission and persistence. In some species, the exosporium forms the outermost structure of the spore and provides the first point of contact between the spore and the environment. The exosporium may also be involved in spore adherence, protection and germination. Clostridium sordellii is a highly lethal, spore forming pathogen that causes soft-tissue infections, enteritis and toxic-shock syndrome. Despite the importance of C. sordellii spores in disease, spore proteins from this bacterium have not been defined or interrogated functionally. In this study, we identified the C. sordellii outer spore proteome and two of the identified proteins, CsA and CsB, were characterised using a genetic and phenotypic approach. Both proteins were essential for the correct formation and positioning of the C. sordellii spore coat and exosporium. The absence of CsA reduced sporulation levels and increased spore sensitivity to heat, sodium hydroxide and hydrochloric acid. By comparison, CsB was required for normal levels of spore adherence to cervical, but not vaginal, cells, with csB mutant spores having increased adherence properties. The establishment of a mouse infection model of the gastrointestinal tract for C. sordellii allowed the role of CsA and CsB to be interrogated in an infected host. Following the oral administration of spores to mice, the wild-type strain efficiently colonized the gastrointestinal tract, with the peak of bacterial numbers occurring at one day post-infection. Colonization was reduced by two logs at four days post-infection. By comparison, mice infected with the csB mutant did not show a reduction in bacterial numbers. We conclude that C. sordellii outer spore proteins are important for the structural and functional integrity of spores. Furthermore, outer spore proteins are required for wild-type levels of colonization during infection, possibly as a result of the role that the proteins play in spore structure and morphology.

Use of Soluble Extracellular Regions of MmpL (SERoM) as Vaccines for Tuberculosis

The current vaccine for tuberculosis (TB) is a live attenuated strain of Mycobacterium bovis (BCG) and while effective at reducing the potential for disseminated TB in young children its disease protection rates in adults is highly variable while it confers little protection against latent TB. With these limitations a new vaccine is desperately needed. We investigated the efficacy of three members of the mycobacterial membrane protein Large (MmpL) family as potential subunit vaccines for TB. MmpLs are large, multifunctional integral membrane proteins, and as such are recalcitrant to purification. Here, we describe a strategy of producing synthetic antigens comprised of the soluble, extracellular regions of MmpL (SERoM)-1, MmpL8 and MmpL10 (SERoM-8 and 10 respectively) as potential vaccine candidates. SERoM-1 and SERoM-8 were determined to be highly immunogenic by IFN-γ ELISpot assays, with 0.1% of all splenocytes from SERoM-1 vaccinated mice producing IFN-γ when re-stimulated with MmpL1. A combined SERoM-1, −8 and −10 vaccine demonstrated significant protection against M. tuberculosis challenge in a murine model of TB, resulting in approximately 10-fold reduction in bacterial numbers following challenge in both the lungs and spleens compared to adjuvant only vaccinated mice. These protective effects were comparable to that achieved with BCG.

Metformin Mediates Protection against Legionella Pneumonia through Activation of AMPK and Mitochondrial Reactive Oxygen Species.

In Legionella pneumophila infection, macrophages play a critical role in the host defense response. Metformin, an oral drug for type 2 diabetes, is attracting attention as a new supportive therapy against a variety of diseases, such as cancer and infectious diseases. The novel mechanisms for metformin actions include modulation of the effector functions of macrophages and other host immune cells. In this study, we have examined the effects of metformin on L. pneumophila infection in vitro and in vivo. Metformin treatment suppressed growth of L. pneumophila in a time- and concentration-dependent fashion in bone marrow-derived macrophages, RAW cells (mouse), and U937 cells (human). Metformin induced phosphorylation of AMP-activated protein kinase (AMPK) in L. pneumophila-infected bone marrow-derived macrophages, and the AMPK inhibitor Compound C negated metformin-mediated growth suppression. Also, metformin induced mitochondrial reactive oxygen species but not phagosomal NADPH oxidase-derived reactive oxygen species. Metformin-mediated growth suppression was mitigated in the presence of the reactive oxygen species scavenger glutathione. In a murine L. pneumophila pneumonia model, metformin treatment improved survival of mice, which was associated with a significant reduction in bacterial number in the lung. Similar to in vitro observations, induction of AMPK phosphorylation and mitochondrial ROS was demonstrated in the infected lungs of mice treated with metformin. Finally, glutathione treatment abolished metformin effects on lung bacterial clearance. Collectively, these data suggest that metformin promotes mitochondrial ROS production and AMPK signaling and enhances the bactericidal activity of macrophages, which may contribute to improved survival in L. pneumophila pneumonia.

Porous Silicon Nanoparticle Delivery of Tandem Peptide Anti-Infectives for the Treatment of Pseudomonas aeruginosa Lung Infections.

There is an urgent need for new materials to treat bacterial infections. In order to improve antibacterial delivery, an anti-infective nanomaterial is developed that utilizes two strategies for localization: i) a biodegradable nanoparticle carrier to localize therapeutics within the tissue, and ii) a novel tandem peptide cargo to localize payload to bacterial membranes. First, a library of antibacterial peptides is screened that combines a membrane-localizing peptide with a toxic peptide cargo and discovers a tandem peptide that displays synergy between the two domains and is able to kill Pseudomonas aeruginosa at sub-micromolar concentrations. To apply this material to the lung, the tandem peptide is loaded into porous silicon nanoparticles (pSiNPs). Charged peptide payloads are loaded into the pores of the pSiNP at ≈30% mass loading and ≈90% loading efficiency using phosphonate surface chemistry. When delivered to the lungs of mice, this anti-infective nanomaterial exhibits improved safety profiles over free peptides. Moreover, treatment of a lung infection of P. aeruginosa results in a large reduction in bacterial numbers and markedly improves survival compared to untreated mice. Collectively, this study presents the selection of a bifunctional peptide-based anti-infective agent and its delivery via biodegradable nanoparticles for application to an animal model of lung infection.

Are antibacterial component additions in etchants and adhesives effective against Streptococcus Mutans?

The aim was to investigate the antibacterial activity of various acids and adhesives with and without antibacterial components against Streptococcus mutans. The antibacterial activities of 35% phosphoric acid (Ultra-Etch), 37% phosphoric acid with benzalkonium chloride (Etch-37), adhesive with chlorhexidine (Peak Universal Bond) and without any agent (PQ1) were investigated by agar-diffusion test. The inhibition-zones were measured after 48 h of incubation. For the tooth-cavity model test; cylindrical cavities were prepared on occlusal dentin surfaces of human molars and divided into four groups (n = 10 cavity/group). Group 1: Ultra-Etch + Peak Universal Bond, Group 2: Ultra-Etch + PQ1, Group 3: Etch-37 + PQ1 were applied. The fourth group without any agent application served as control. The teeth were immersed in 5.8 × 106 cfu/ml of S. mutans solution to infect the cavities for 72 h before the application of the groups. After 72 h, dentin chips were collected from the cavity walls with burs for bacterial counting. Statistical analysis was performed by ANOVA, Bonferroni and Dunnett C tests (p < 0.05). Ultra-Etch and Etch-37 performed similar antibacterial activities in agar-diffusion test. Both acids showed better antibacterial activity compared to adhesives (p < 0.05). The antibacterial activity of PQ1 and Peak Universal Bond was observed to be inactivated by light-polymerization. According to the tooth-cavity model; Group I, II, and III demonstrated reduction in bacterial number and there was no significant difference between them. Antibacterial component additions in etchant and adhesive did not show superior antibacterial activity against S. mutans in both in vitro tests.

In vitro and In vivo Activity of Theaflavin-Epicatechin Combinations versus Multidrug-Resistant Acinetobacter baumannii.

Introduction Acinetobacter baumannii is an important human nosocomial pathogen; most clinical isolates are multidrug-resistant (MDR). Infections caused by A. baumannii often lead to high morbidity and mortality, with limited treatment options. Owing to the small number of anti-Gram-negative antibiotics in the development pipeline, researchers are looking to other natural compounds. The aim of this study was to determine the in vitro kill kinetics, in vivo efficacy and toxicity of theaflavin–epicatechin combinations against MDR A. baumannii.MethodsKill-kinetic assays were performed in Mueller–Hinton 2 broth over 24 h. Toxicity of the compound in the insect model, Galleria mellonella was investigated. The effect of theaflavin–epicatechin combinations on mortality and morbidity were assessed in Acinetobacter baumannii-infected G. mellonella. Larvae were scored for morbidity (melanisation: scale; 0–4) and mortality over 96 h.ResultsKill-kinetic assays revealed that monotherapy had bacteriostatic activity over 24 h, whereas theaflavin–epicatechin combinations were bactericidal (a >3 log reduction in bacterial numbers at 24 h compared with the starting inoculum). Both polyphenols were non-toxic to G. mellonella at concentrations of up to 1000 mg/kg. In vivo treatment assays showed that the combination significantly increased (t test; p ≤ 0.05) larval survival at 96 h to 86% [±17 standard deviation percentage points (pp)] compared to monotherapy with theaflavin (52% ± 14 pp), epicatechin (44% ± 25 pp) or PBS (31% ± 13 pp). Morbidity was also lower in larvae treated with the combination, compared with monotherapy.ConclusionPolyphenol combinations produce effective antibacterial action against A. baumannii and show great potential for the treatment of infections caused by MDR A. baumannii.

Novel vaccine potential of Rv3131, a DosR regulon-encoded putative nitroreductase, against hyper-virulent Mycobacterium tuberculosis strain K

Accumulating evidence indicates that latency-associated Mycobacterium tuberculosis (Mtb)-specific antigens from the dormancy survival regulator regulon (DosR) may be promising novel vaccine target antigens for the development of an improved tuberculosis vaccine. After transcriptional profiling of DosR-related genes in the hyper-virulent Beijing Mtb strain K and the reference Mtb strain H37Rv, we selected Rv3131, a hypothetical nitroreductase, as a vaccine antigen and evaluated its vaccine efficacy against Mtb K. Mtb K exhibited stable and constitutive up-regulation of rv3131 relative to Mtb H37Rv under three different growth conditions (at least 2-fold induction) including exponential growth in normal culture conditions, hypoxia, and inside macrophages. Mice immunised with Rv3131 formulated in GLA-SE, a well-defined TLR4 adjuvant, displayed enhanced Rv3131-specific IFN-γ and serum IgG2c responses along with effector/memory T cell expansion and remarkable generation of Rv3131-specific multifunctional CD4+ T cells co-producing TNF-α, IFN-γ and IL-2 in both spleen and lung. Following challenge with Mtb K, the Rv3131/GLA-SE-immunised group exhibited a significant reduction in bacterial number and less extensive lung inflammation accompanied by the obvious persistence of Rv3131-specific multifunctional CD4+ T cells. These results suggest that Rv3131 could be an excellent candidate for potential use in a multi-antigenic Mtb subunit vaccine, especially against Mtb Beijing strains.

Antibacterial activity of resin composites containing surface pre-reacted glass-ionomer (S-PRG) filler

ObjectiveA surface pre-reacted glass-ionomer (S-PRG) filler is a technology of interest for providing bio-functions to restorative materials. Resin composites containing S-PRG filler have been reported to show less plaque accumulation and reduced bacterial attachment. In this study, experimental resin composites containing S-PRG filler at various concentrations were fabricated, and the inhibitory effects on bacterial growth on their surface and the association of ions released from S-PRG filler with antibacterial activity were evaluated. MethodsFive kinds of experimental resin composites containing S-PRG filler at 0, 13.9, 27.3, 41.8, or 55.9 (vol.%) were fabricated. Streptococcus mutans was cultured on the cured discs for 18h to examine the growth of bacteria in contact with the surface of the experimental resins. The concentrations of Al3+, BO33−, F−, Na+, SiO32−, or Sr2+ released from each experimental resin into water were measured. The standardized solutions of each ion were prepared at the concentrations determined to be released from the experimental resin, and their inhibitory effects of single ion species on S. mutans growth were evaluated by using each standardized solution. ResultsResin composites containing S-PRG filler at 13.9 (vol.%) or greater inhibited S. mutans growth on their surface. When S. mutans was incubated in the presence of six kinds of ions at the concentrations released from the resin composite containing S-PRG filler at 55.9 (vol.%), a significant reduction in bacterial number was observed for BO33−, F−, Al3+, and SiO32−. Among these four ions, BO33− and F− demonstrated the strongest inhibitory effect on S. mutans growth. SignificanceOur findings suggest that resin composites containing S-PRG filler inhibit the growth of S. mutans on their surface. BO33−, F−, Al3+ and SiO32− released from S-PRG filler have the ability to inhibit S. mutans growth, and the inhibitory effects are mainly attributed to release of BO33− and F−.

Antimicrobial activity of a bioelectric dressing using an in vitro wound pathogen colony drip-flow reactor biofilm model.

We performed in vitro antibiofilm testing of a silver and zinc containing microcurrent generating bioelectric dressing (BED) against clinical wound pathogens to determine its efficacy in preventing biofilm formation under low shear and continuous flow conditions, simulating wound infection environments. We customised an in vitro colony drip-flow reactor (DFR) biofilm model for efficacy evaluation of BED. Each bacterial pathogen was diluted to 104CFU/ml and inoculated on the polycarbonate filter membrane as an abiotic support. BED and controls (no treatment, gauze, and blank polyester with no silver and zinc) were applied directly on the membranes where bacterial cultures were inoculated. Biofilms were continuously developed onto the membranes for 72 hours at room temperature. Biofilm formation was confirmed by crystal violet staining and microscopic observation. Through vigorous shaking and sonication, the released bacteria were serially diluted, plated, and incubated for 24 hours at 37°C to determine the numbers of surviving bacteria. Biofilms were well developed onto blank polyesters, but not the BED after 72 hours incubation. Crystal violet staining from the blank polyesters showed large and fully grown biofilms. We observed an inhibition in bacterial growth on BED treatment. The antibiofilm activity of the BED against each of eight monospecies biofilms showed a 1- or 2 log10 (or 10- or 100-fold) reduction in bacterial numbers compared with those of controls. Our results demonstrated that colony DFR biofilm model was an appropriate for testing the antibiofilm efficacy of BED under low shear and continuous flow conditions for simulating clinical wound environments. The bioelectric currents generated from the silver and zinc active ingredients in the dressing resulted in antibiofilm activity of this wound care device. The opinions or assertions contained herein are the private views of the authors, based on scientific investigation, and are not to be construed as official or as reflecting the views of the Department of Defense, the United States Government or any of the authors' employers. Dr. Mina Izadjoo has served as a consultant to Vomaris Innovations Inc.

The bactericidal activity of glutaraldehyde-impregnated polyurethane.

Although glutaraldehyde is known to be bactericidal in solution, its potential use to create novel antibacterial polymers suitable for use in healthcare environments has not been evaluated. Here, novel materials were prepared in which glutaraldehyde was either incorporated into polyurethane using a simple “swell‐encapsulation‐shrink” method (hereafter referred to as “glutaraldehyde‐impregnated polyurethane”), or simply applied to the polymer surface (hereafter referred to as “glutaraldehyde‐coated polyurethane”). The antibacterial activity of glutaraldehyde‐impregnated and glutaraldehyde‐coated polyurethane samples was tested against Escherichia coli and Staphylococcus aureus. Glutaraldehyde‐impregnated polyurethane resulted in a 99.9% reduction in the numbers of E. coli within 2 h and a similar reduction of S. aureus within 1 h, whereas only a minimal reduction in bacterial numbers was observed when the biocide was bound to the polymer surface. After 15 days, however, the bactericidal activity of the impregnated material was substantially reduced presumably due to polymerization of glutaraldehyde. Thus, although glutaraldehyde retains antibacterial activity when impregnated into polyurethane, activity is not maintained for extended periods of time. Future work should examine the potential of chemical modification of glutaraldehyde and/or polyurethane to improve the useful lifespan of this novel antibacterial polymer.

Copper-based water repellent and antibacterial coatings by aerosol assisted chemical vapour deposition.

The adhesion and proliferation of bacteria on solid surfaces presents a major challenge in both healthcare and industrial applications. In response to this problem, an effective and simple method is reported to fabricate superhydrophobic antibacterial copper coated polymer films via aerosol assisted chemical vapor deposition (AACVD). The material is characterized using a range of techniques including electron microscopy, water contact angle measurement and elemental mapping. The antibacterial activity of the modified film is tested against the Gram-negative bacterium, Escherichia coli, and the Gram-positive bacterium, Staphylococcus aureus and the film shows highly significant antibacterial activity against both bacteria (>4 log reduction in bacterial numbers) in 15 min and 60 min, respectively. In addition, all the CVD modified samples results in a significant reduction in bacterial cell adhesion compared to the control materials. Thus, we report a new film type that has dual mode of action-the superhydrophobicity helps limit cell adhesion combined with a cytotoxic copper induced bacteria kill.

Egg surface decontamination with bronopol increases larval survival of Nile tilapia, Oreochromis niloticus

Intensive tilapia egg incubation techniques create favourable conditions for microbial proliferation which often leads to massive mortalities of fish larvae. The effects of Nile tilapia (Oreochromis niloticus) eggs exposure to bronopol on decontamination of their bacterial surface and survival of larvae were observed. Immersion treatments of fertilized eggs were applied at 10, 25, 50, 100, and 250 mg/l of bronopol for 10, 20, and 30 min. This treatment substantially reduced the number of bacteria on Nile tilapia eggs. The greatest reduction in bacterial numbers (1.58 × 103 colony forming units/g of egg) was observed at the maximum treatment dosage, i.e. 250 mg/l for 30 min, but this was not significant when compared with treatments of 100-250 mg/l bronopol for 10-30 min. Treatments of 50-250 mg/l bronopol provided better larval survival (89.33-94.67%) than those of < 50 mg/l (P < 0.05). The larval survival obtained from the Nile tilapia eggs disinfected with 10 and 25 mg/l bronopol for 10-30 min was similar to that of negative controls (79.33-80.97%). Additional in vitro test was performed to determine the inhibitory potency of bronopol against bacteria cultured from fertilized Nile tilapia eggs. Twenty-two bacterial isolates (Aeromonas hydrophila (n = 12), Staphylococcus aureus (n = 6), Escherichia coli (n = 2), and Micrococcus spp. (n = 2)) responded to minimum inhibitory concentrations of bronopol ranging from 64 to 128 µg/ml. The study demonstrates that the immersion treatment of Nile tilapia fertilized eggs with 250 mg/l for 10-30 min, the concentration proven effective against bacteria cultured from fertilized eggs, significantly reduced bacterial load and improved larval survival.

Bio-Control of Salmonella Enteritidis in Foods Using Bacteriophages.

Two lytic phages, vB_SenM-PA13076 (PA13076) and vB_SenM-PC2184 (PC2184), were isolated from chicken sewage and characterized with host strains Salmonella Enteritidis (SE) ATCC13076 and CVCC2184, respectively. Transmission electron microscopy revealed that they belonged to the family Myoviridae. The lytic abilities of these two phages in liquid culture showed 104 multiplicity of infection (MOI) was the best in inhibiting bacteria, with PC2184 exhibiting more activity than PA13076. The two phages exhibited broad host range within the genus Salmonella. Phage PA13076 and PC2184 had a lytic effect on 222 (71.4%) and 298 (95.8%) of the 311 epidemic Salmonella isolates, respectively. We tested the effectiveness of phage PA13076 and PC2184 as well as a cocktail combination of both in three different foods (chicken breast, pasteurized whole milk and Chinese cabbage) contaminated with SE. Samples were spiked with 1 × 104 CFU individual SE or a mixture of strains (ATCC13076 and CVCC2184), then treated with 1 × 108 PFU individual phage or a two phage cocktail, and incubated at 4 °C or 25 °C for 5 h. In general, the inhibitory effect of phage and phage cocktail was better at 4 °C than that at 25 °C, whereas the opposite result was observed in Chinese cabbage, and phage cocktail was better than either single phage. A significant reduction in bacterial numbers (1.5–4 log CFU/sample, p < 0.05) was observed in all tested foods. The two phages on the three food samples were relatively stable, especially at 4 °C, with the phages exhibiting the greatest stability in milk. Our research shows that our phages have potential effectiveness as a bio-control agent of Salmonella in foods.

Ozonated saline shows activity against planktonic and biofilm growing Staphylococcus aureus in vitro: a potential irrigant for infected wounds.

Infections associated with deep wounds require extensive surgical and medical care. New adjunctive treatments are required to aid in the eradication of the bacterial biofilms found on infected wounds and, in particular, any underlying hardware. Ozone has been used as a safe and efficient disinfectant in water treatment plants for many years. The purpose of this study is to investigate the anti-biofilm potential of ozonated saline against biofilms of Staphylococcus aureus, a microorganism commonly implicated in wound infections. A custom-made bacterial biofilm bioreactor was used to grow S. aureus biofilms on discs of medical grade titanium alloy. An ozone generator was connected in-line and biofilms and planktonic bacteria were exposed to ozone in saline. Cytotoxicity was assessed against primary ovine osteoblasts in the same system. In tests against planktonic S. aureus, a 99% reduction in bacterial numbers was detected within 15 minutes of exposure. S. aureus biofilms were significantly more resistant to ozone, although complete eradication of the biofilm was eventually achieved within 5 hours. Ozonated saline was not found to be cytotoxic to primary ovine osteoblasts. Ozonated saline may be suitable as an adjuvant therapy to treat patients as an instillation fluid for wound irrigation and sterilisation.

Application of quantitative microbial risk assessment for selection of microbial reduction targets for hard surface disinfectants

This quantitative microbial risk assessment (QMRA) included problem formulation for fomites and hazard identification for 7 microorganisms, including pathogenic Escherichia coli and E coli 0157:H7, Listeria monocytogenes, norovirus, Pseudomonas spp, Salmonella spp, and Staphylococcus aureus. The goal was to address a risk-based process for choosing the log10 reduction recommendations, in contrast to the current US Environmental Protection Agency requirements. For each microbe evaluated, the QMRA model included specific dose-response models, occurrence determination of aerobic bacteria and specific organisms on fomites, exposure assessment, risk characterization, and risk reduction. Risk estimates were determined for a simple scenario using a single touch of a contaminated surface and self-inoculation. A comparative analysis of log10 reductions, as suggested by the US Environmental Protection Agency, and the risks based on this QMRA approach was also undertaken. The literature review and meta-analysis showed that aerobic bacteria were the most commonly studied on fomites, averaging 100 colony-forming units (CFU)/cm(2). Pseudomonas aeruginosa was found at a level of 3.3 × 10(-1) CFU/cm(2); methicillin-resistant S aureus (MRSA), at 6.4 × 10(-1) CFU/cm(2). Risk estimates per contact event ranged from a high of 10(-3) for norovirus to a low of 10(-9) for S aureus. This QMRA analysis suggests that a reduction in bacterial numbers on a fomite by 99% (2 logs) most often will reduce the risk of infection from a single contact to less than 1 in 1 million.

Laboratory evaluation of Drawtex Hydroconductive Dressing with LevaFiber technology

The aim of this study was to assess absorption, sequestration and retention of microorganisms by Drawtex Hydroconductive dressing with LevaFiberTM technology. The absorption over time and the ability to sequester and retain bacteria were assessed in the laboratory using an in vitro model where known amounts of fluid and microorganisms were allowed to absorb and sequester over a 24 hour period. The reduction in numbers of microorganisms following sequestration was determined using standard plate counting methods. Retention of the organisms was visualised by scanning electron microscopy. Drawtex was shown to absorb eight times its own weight in fluid over time and it showed a 90% reduction in bacterial numbers over a 24 hour period in sequestration experiments. Direct observation by scanning electron microscopy demonstrated bacterial retention in the dressing fibres. Drawtex is a recent addition to the formulary for absorbent dressings available in the UK. It demonstrates excellent absorption properties, while maintaining integrity. In addition, it sequesters and retains microorganisms, which will help with removal of exudate and bioburden from the wound bed to help facilitate wound healing. This study was funded by an educational grant from Martindale Pharma, UK. The authors have no conflicts of interest to declare with regard to the manuscript or its content.

Development of a controlling method for Escherichia coli O157:H7 and Salmonella spp. in fresh market beef by using polylysine and modified atmosphere packaging

Escherichia coli O157:H7 and Salmonella spp. often contaminate fresh beef. In Japan, an E coli outbreak caused by raw beef made 181 people ill and 5 individuals dead in 2011. Responding to this outbreak, an effective sterilization method for fresh beef is expected to be developed. In this study, ε-polylysine combined with CO2-packaging method was examined for controlling these pathogens in fresh beef. At an incubation temperature of 4 °C, approximately 4.3 log and 2.4 log reduction in bacterial numbers were observed after 7-day incubation for E. coli O157:H7 and Salmonella, respectively, in ε-polylysine-added beef. When effectiveness of CO2-packaging combined with ε-polylysine was investigated, CO2 did not have additional inhibiting effect on bacterial growth compared to only-ε-polylysine-treated samples when incubated at 4 °C. However, effectiveness of CO2 was observed when incubated at 10 °C where approximately 2.9 log and 4.4 log reduction in E. coli cell numbers were observed in only-ε-polylysine-treated samples and polylysine- and CO2-treated group, respectively, and approximately 1.7 log and 3.5 log reduction in Salmonella cell numbers were observed in only-ε-polylysine-treated samples and polylysine and CO2-treated group, respectively. This study confirmed that ε-polylysine or ε-polylysine combined with CO2 packaging are effective in preventing foodborne diseases caused by raw beef.