Log10 Reduction In Viability Research Articles

Is a reduction in viability enough to determine biofilm susceptibility to a biocide?

The abundance and prevalence of dry-surface biofilms (DSBs) in hospitals constitute an emerging problem, yet studies rarely report the cleaning and disinfection efficacy against DSBs. Here, the combined impact of treatments on viability, transferability, and recovery of bacteria from DSBs has been investigated for the first time. Staphylococcus aureus DSBs were produced in alternating 48-hour wet-dry cycles for 12 days on AISI 430 stainless steel discs. The efficacy of 11 commercially available disinfectants, 4 detergents, and 2 contactless interventions were tested using a modified standardized product test. Reduction in viability, direct transferability, cross transmission (via glove intermediate), and DSB recovery after treatment were measured. Of 11 disinfectants, 9 were effective in killing and removing bacteria from S. aureus DSBs with >4 log10 reduction. Only 2 disinfectants, sodium dichloroisocyanurate 1,000 ppm and peracetic acid 3,500 ppm, were able to lower both direct and cross transmission of bacteria (<2 compression contacts positive for bacterial growth). Of 11 disinfectants, 8 could not prevent DSB recovery for >2 days. Treatments not involving mechanical action (vaporized hydrogen peroxide and cold atmospheric plasma) were ineffective, producing <1 log10 reduction in viability, DSB regrowth within 1 day, and 100% transferability of DSB after treatment. Reduction in bacterial viability alone does not determine product performance against biofilm and might give a false sense of security to consumers, manufacturers and regulators. The ability to prevent bacterial transfer and biofilm recovery after treatment requires a better understanding of the effectiveness of biocidal products.

It's a trap! The development of a versatile drain biofilm model and its susceptibility to disinfection

Pathogens in drain biofilms pose a significant risk for hospital-acquired infection. However, the evidence of product effectiveness in controlling drain biofilm and pathogen dissemination are scarce. A novel in-vitro biofilm model was developed to address the need for a robust, reproduceable and simple testing methodology for disinfection efficacy against a complex drain biofilm. Identical complex drain biofilms were established simultaneously over 8 days, mimicking a sink trap. Reproducibility of their composition was confirmed by next-generation sequencing. The efficacy of sodium hypochlorite 1000 ppm (NaOCl), sodium dichloroisocyanurate 1000 ppm (NaDCC), non-ionic surfactant (NIS) and peracetic acid 4000 ppm (PAA) was explored, simulating normal sink usage conditions. Bacterial viability and recovery following a series of 15-min treatments were measured in three distinct parts of the drain. The drain biofilm consisted of 119 mixed species of Gram-positive and -negative bacteria. NaOCl produced a >4 log10 reduction in viability in the drain front section alone, while PAA achieved a >4 log10 reduction in viability in all of the drain sections following three 15-min doses and prevented biofilm regrowth for >4 days. NIS and NaDCC failed to control the biofilm in any drain sections. Drains are one source of microbial pathogens in healthcare settings. Microbial biofilms are notoriously difficult to eradicate with conventional chemical biocidal products. The development of this reproducible in-vitro drain biofilm model enabled understanding of the impact of biocidal products on biofilm spatial composition and viability in different parts of the drain.

Use of a small‐scale, portable test chamber for determining the bactericidal efficacy of aerosolized glycol formulations

This study aimed to understand the efficacy and mechanisms of action of an aerosolised glycol-ethanol formulations against bacteria. We validated a small scale in-house test chamber to determine the microbicidal efficacy of four aerosolized formulations combining dipropylene glycol and ethanol against Staphylococcus aureus and Escherichia coli embedded in alginate. The aerosolised glycol/ethanol formulation decreased bacterial viability by 3 Log10 and was more efficacious than an ethanol only control formulation. Electron microscopic examination indicated extensive structural damage in both bacteria, and membrane damage was confirmed with potassium release in S. aureus and DNA release in E. coli. The development of a small test chamber facilitated the measurement of the microbicidal efficacy and experiments to understand the mechanism of action of an aerosolised microbicidal formulation.

Candida auris Dry Surface Biofilm (DSB) for Disinfectant Efficacy Testing.

Candida auris is an emerging pathogen that needs to be controlled effectively due to its association with a high mortality rate. The presence of biofilms on dry surfaces has been shown to be widespread in healthcare settings. We produced a C. auris dry surface biofilm (DSB) on stainless steel surfaces following sequential hydration and desiccation cycles for 12 days. The ASTM2967-15 was used to measure the reduction in viability of 12 commercially wipe-based disinfectants and sodium hypochlorite (1000 ppm) against C. auris DSB. We also evaluated C. auris transferability and biofilm regrowth post-treatment. A peracetic acid (3500 ppm) product and two chlorine-based products (1000 ppm available chlorine) were successful in reducing C. auris viability and delaying DSB regrowth. However, 50% of the products tested failed to decrease C. auris viability, 58% failed to prevent its transferability, and 75% did not delay biofilm regrowth. Using three different parameters to measure product efficacy provided a practical evaluation of product effectiveness against C. auris DSB. Although log10 reduction in viability is traditionally measured, transferability is an important factor to consider from an infection control and prevention point of view as it allows for determination of whether the surface is safe to touch by patients and hospital staff post-treatment.

Antimicrobial efficacy of a novel povidone iodine contact lens disinfection system.

Contact lens (CL) wear is a risk factor for the acquisition of microbial keratitis. Accordingly, compliance to manufacturers' recommended hygiene and disinfection procedures are vital to safe (CL) use. In this study we evaluated a novel povidone-iodine (PI) (CL) disinfection system (cleadew, Ophtecs Corporation, Japan) against a range of bacterial, fungal and Acanthamoeba. Antimicrobial assays were conducted according to ISO 14729 using the recommended strains of bacteria and fungi, with and without the presence of organic soil. Regrowth of bacteria and fungi in the disinfection system was also examined. The activity on biofilms formed from Stenotrophomonas maltophilia and Achromobacter sp. was evaluated. Efficacy against A. castellanii trophozoites and cysts was also investigated. The PI system gave >4 log10 kill of all bacteria and fungi following the manufacturer's recommended disinfection and cleaning time of 4h, with or without the presence of organic soil. No regrowth of organisms was found after 14days in the neutralized solution. In the biofilm studies the system resulted in at least a 7 log10 reduction in viability of bacteria. For Acanthamoeba, >3 log10 kill of trophozoites and 1.1-2.8 log10 kill for the cyst stage was obtained. The PI system effective against a variety of pathogenic microorganisms under a range of test conditions. Strict compliance to recommended CL hygiene procedures is essential for safe CL wear. The use of care systems such as PI, with broad spectrum antimicrobial activity, may aid in the prevention of potentially sight threatening microbial keratitis.

The effect of hospital biocide sodium dichloroisocyanurate on the viability and properties of Clostridium difficile spores.

This study is the first to report on changes in Clostridium difficile spore surface property after exposure to sublethal levels of the commonly used biocide sodium dichloroisocyanurate. The implications of these changes to the spore surface include increased adherence of the spores to inorganic surfaces which can directly contribute to persistence and spread of spores within the hospital environment.

Biocompatible biodegradable polymeric antibacterial nanoparticles for enhancing the effects of a third-generation cephalosporin against resistant bacteria.

In the present study, enhancement of the the antibacterial activity of ceftriaxone against Gram-positive (meticillin-resistant Staphylococcus aureus; MRSA) and Gram-negative (Escherichia coli) bacteria with a biodegradable polymer was attempted. MRSA and E. coli were collected and identified by biochemical and molecular tests. Blank and ceftriaxone-loaded chitosan nanoparticles (CNPs) were prepared by the ionic gelation method. In vitro antibiotic-susceptibility studies were performed by disc diffusion, agar well plate method, Etest and time-kill assay. In vivo activity was assessed using the neutropenic mouse thigh model and cytotoxicity was estimated by MTT (methylthiazolyldiphenyl tetrazolium bromide) assay with the MCF-7 cancer cell line. MRSA showed 97 % and E. coli 83 % resistance against ceftriaxone in the disc diffusion test. The isolates showing a ≥1024 mg l-1 MIC value for ceftriaxone were selected for further evaluation. In the agar well plate method, the mean zones of inhibition for blank and ceftriaxone-loaded CNPs were 17 and 23 mm, respectively, for MRSA isolates and 15 and 25 mm, respectively, for E. coli isolates. In the time-kill assay, ~1 log10 to ~2.5 log10 reduction in viability was seen with both isolates when treated with ceftriaxone-loaded CNPs over 24 h. The in vivo studies also showed the enhanced antibacterial activity of ceftriaxone-loaded CNPs, with a 41 % reduction in MRSA and a 27 % reduction in E. coli burden. A low cytotoxicity of blank and ceftriaxone-loaded CNPs was seen, with a slight reduction in the percentage viability of cells from 87 to 83 % and from 88 to 81 %, respectively. The synergistic effect of ceftriaxone-loaded CNPs is a useful finding for the treatment of MRSA and E. coli infections.

Predation Efficacy of Bdellovibrio bacteriovorus on Multidrug-Resistant Clinical Pathogens and Their Corresponding Biofilms.

The aim of the present study was to evaluate the predation efficacy of Bdellovibrio bacteriovorus on multidrug-resistant (MDR) or extensive drug resistant (XDR) gram-negative pathogens and their corresponding biofilms. In this study, we examined the ability of B. bacteriovorus to prey on MDR and XDR gram-negative clinical bacteria, including Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Results showed that B. bacteriovorus was able to prey on all planktonic cultures, among which the most efficient predation was observed for drug-resistant E. coli, with a 3.11 log10 reduction in viability. Furthermore, B. bacteriovorus demonstrated promising efficacy in preventing biofilm formation and dispersing the established biofilm. Reductions in biofilm formation of E. coli, K. pneumoniae, P. aeruginosa, and A. baumannii co-cultured with B. bacteriovorus were 65.2%, 37.1%, 44.7%, and 36.8%, respectively. Meanwhile, the established biofilms of E. coli, K. pneumoniae, P. aeruginosa, and A. baumannii were significantly reduced by 83.4%, 81.8%, 83.1%, and 79.9%, respectively. A visual analysis supported by scanning electron microscopy demonstrated the role of B. bacteriovorus in removing the established biofilms. This study highlights the potential use of B. bacteriovorus as a biological control agent with the capability to prey on MDR/XDR gram-negative pathogens and eradicate biofilms.

Antimicrobial activity of novel nanostructured Cu-SiO2 coatings prepared by chemical vapour deposition against hospital related pathogens

There is increasing recognition that the healthcare environment acts as an important reservoir for transmission of healthcare acquired infections (HCAI). One method of reducing environmental contamination would be use of antimicrobial materials. The antimicrobial activity of thin silica-copper films prepared by chemical vapour deposition was evaluated against standard strains of bacteria used for disinfectant testing and bacteria of current interest in HCAI. The structure of the coatings was determined using Scanning Electron Microscopy and their hardness and adhesion to the substrate determined. Antimicrobial activity was tested using a method based on BS ISO 22196:2007. The coatings had a pale green-brown colour and had a similar hardness to steel. SEM showed nano-structured aggregates of Cu within a silica matrix. A log10 reduction in viability of >5 could be obtained within 4 h for the disinfectant test strains and within 6 h for producing Acinetobacter baumannii, Klebsiella pneumoniae and Stenotrophomonas maltophilia. Activity against the other hospital isolates was slower but still gave log10 reduction factors of >5 for extended spectrum β-lactamase producing Escherichia coli and >3 for vancomycin resistant Enterococcus faecium, methicillin resistant Staphylococcus aureus and Pseudomonas aeruginosa within 24 h. The results demonstrate the importance of testing antimicrobial materials destined for healthcare use against isolates of current interest in hospitals as well as standard test strains. The coatings used here can also be applied to substrates such as metals and ceramics and have potential applications where reduction of microbial environmental contamination is desirable.

Role of the competitive microbial flora in the radiation-induced enhancement of ochratoxin production by Aspergillus alutaceus var. alutaceus NRRL 3174

The radiation sensitivity and the toxigenic potential of conidiospores of the fungus Aspergillus alutaceus var. alutaceus were determined after irradiation with 60Co gamma rays and high-energy electrons. Over the pH range of 3.6 to 8.8, the doses required for a 1 log10 reduction in viability based on the exponential portion of the survival curve ranged from 0.21 to 0.22 kGy, with extrapolation numbers (extrapolation of the exponential portion of the survival curve to zero dose) of 1.01 to 1.33, for electron irradiation, and from 0.24 to 0.27 kGy, with extrapolation numbers of 2.26 to 5.13, for gamma irradiation. Nonsterile barley that was inoculated with conidia of the fungus and then irradiated with either electrons or gamma rays and incubated for prolonged periods at 28 degrees C and at a moisture content of 25% produced less ochratoxin A with increasing doses of radiation. Inoculation of barley following irradiation resulted in enhanced ochratoxin levels compared with unirradiated controls. In these experiments, inoculation with 10(2) spores per g produced greater radiation-induced enhancement than inoculation with 10(5) spores per g. There was no radiation-induced enhancement when the barley was surface sterilized by chemical means prior to irradiation. These results are consistent with the hypothesis that a reduction in the competing microbial flora by irradiation is responsible for the enhanced mycotoxin production observed when nonsterile barley is inoculated with the toxigenic fungus A. alutaceus var. alutaceus after irradiation.

Oral Multivitamin Liquid Preparation

Contact lens (CL) wear is a risk factor for the acquisition of microbial keratitis. Accordingly, compliance to manufacturers’ recommended hygiene and disinfection procedures are vital to safe (CL) use. In this study we evaluated a novel povidone-iodine (PI) (CL) disinfection system (cleadew, Ophtecs Corporation, Japan) against a range of bacterial, fungal and Acanthamoeba.Antimicrobial assays were conducted according to ISO 14729 using the recommended strains of bacteria and fungi, with and without the presence of organic soil. Regrowth of bacteria and fungi in the disinfection system was also examined. The activity on biofilms formed from Stenotrophomonas maltophilia and Achromobacter sp. was evaluated. Efficacy against A. castellanii trophozoites and cysts was also investigated.The PI system gave >4 log10 kill of all bacteria and fungi following the manufacturer’s recommended disinfection and cleaning time of 4 h, with or without the presence of organic soil. No regrowth of organisms was found after 14 days in the neutralized solution. In the biofilm studies the system resulted in at least a 7 log10 reduction in viability of bacteria. For Acanthamoeba, >3 log10 kill of trophozoites and 1.1–2.8 log10 kill for the cyst stage was obtained.The PI system effective against a variety of pathogenic microorganisms under a range of test conditions. Strict compliance to recommended CL hygiene procedures is essential for safe CL wear. The use of care systems such as PI, with broad spectrum antimicrobial activity, may aid in the prevention of potentially sight threatening microbial keratitis.