Epidemiological evidence suggests that patients admitted to rooms previously occupied by individuals infected with multidrug-resistant organisms (MDROs) face an increased risk of acquiring these pathogens. This raises the question of whether MDROs also exhibit resistance to used disinfectants. This article reviews the current state of research on this topic.

The spread of multidrug-resistant bacteria from humans or livestock is a critical issue. However, the epidemiology of resistant pathogens across wastewater pathways is poorly understood. Therefore, we performed a detailed comparison of third-generation cephalosporin-resistant Escherichia coli (3GCREC) from wastewater treatment plants (WWTPs) to analyze dissemination pathways. A total of 172 3GCREC isolated from four WWTPs were characterized via whole genome sequencing. Clonal relatedness was determined using multi-locus sequence typing (MLST) and core genome MLST. Resistance genotypes and plasmid replicons were determined. A total of 68 MLST sequence types were observed with 28 closely related clusters. Resistance genes to eight antibiotic classes were detected. In fluoroquinolone-resistant isolates, resistance was associated with three-or-more point mutations in target genes. Typing revealed high genetic diversity with only a few clonal lineages present in all WWTPs. The distribution paths of individual lines could only be traced in exceptional cases with a lack of enrichment of certain lineages. Varying resistance genes and plasmids, as well as fluoroquinolone resistance-associated point mutations in individual isolates, further corroborated the high diversity of 3GCREC in WWTPs. In total, we observed high diversity of 3GCREC inside the tested WWTPs with proof of resistant strains being released into the environment even after treatment processes.

Epidemiological studies show that the care of patients in rooms with a previous stay by a person with evidence of multi-resistant pathogens (MRP) is associated with an increased risk of these pathogens occurring. The question therefore regularly arises as to whether MRP also exhibit resistance to the disinfectants used. To date, there are no standardised definitions for "resistance" to disinfectants. However, disinfectants authorised on the market are also effective against multi-resistant pathogens and the failure of efficient disinfection is mainly caused by application errors (insufficient cleaning, incomplete wetting, incorrect application concentration or exposure time etc.). The effectiveness of disinfectants depends on a variety of environmental factors (especially accompanying contamination). A reduced sensitivity to disinfectants can occur in individual isolates due to selection under sub-inhibitory concentrations of disinfectants. Resistance mechanisms to antibiotics do not mediate cross-resistance to disinfectants, but a change in the permeability of bacterial cells can influence sensitivity to disinfectants and antibiotics. In general, the success of routine disinfection can be improved by suitable process controls and contribute to reducing the transmission of MRP.

Epidemiological studies show that the care of patients in rooms with a previous stay by a person with evidence of multi-resistant pathogens (MRP) is associated with an increased risk of these pathogens occurring. The question therefore regularly arises as to whether MRP also exhibit resistance to the disinfectants used. To date, there are no standardised definitions for "resistance" to disinfectants. However, disinfectants authorised on the market are also effective against multi-resistant pathogens and the failure of efficient disinfection is mainly caused by application errors (insufficient cleaning, incomplete wetting, incorrect application concentration or exposure time etc.). The effectiveness of disinfectants depends on a variety of environmental factors (especially accompanying contamination). A reduced sensitivity to disinfectants can occur in individual isolates due to selection under sub-inhibitory concentrations of disinfectants. Resistance mechanisms to antibiotics do not mediate cross-resistance to disinfectants, but a change in the permeability of bacterial cells can influence sensitivity to disinfectants and antibiotics. In general, the success of routine disinfection can be improved by suitable process controls and contribute to reducing the transmission of MRP.

BACKGROUND:Antimicrobial materials or surfaces are advertised as part of infection prevention bundles. However, the efficacy of such antimicrobial surfaces has not been sufficiently investigated in hospitals. In this study, the antimicrobial activity of examination gloves with light-activated antimicrobial properties against Gram-positive microorganisms was investigated modelling real live conditions. METHOD:In a standardized experimental set-up with dry and realistic contamination, the antimicrobial properties of gloves claiming light dependent antimicrobial activity against Gram-positive organisms were tested in comparison with conventional examination gloves. All gloves were contaminated through a standardized activity of the test persons for construction with contaminated building blocks. For contamination suspensions of Enterococcus faecium ATCC 6057, Acinetobacter baumannii (outbreak strain), methicillin resistant Staphylococcus aureus ATCC 43300 or E. faecium (VRE) patient isolate were dried on the surfaces. After the standardized activity, the gloves were held for 10 min in the light present in the room (bright conditions) and the grade of contamination was determined subsequently by quantitative culture. In one experimental series gloves were held in a dark box after contamination as a control (dark conditions). RESULTS:The light intensity in all experiments under bright conditions was significantly above the limit value specified by the manufacturer for the activation of antimicrobial properties (> 500 lx). The mean values for experiments with antimicrobial active and non-active gloves were 955 and 935 lx, respectively. As claimed by the manufacture, the gloves showed no sufficient efficacy against A. baumannii under bright conditions. Against Gram-positive microorganisms such as E. faecium, E. faecium (VRE) and methicillin resistant S. aureus the gloves showed only very low antimicrobial activity with a reduction factor < 1 log10 even after 10 min in bright conditions. Interestingly, comparable results for experiments with A. baumannii and E. faecium were shown under dark conditions. CONCLUSION:The lack of activity of the active principle against Gram-negative microorganisms could be confirmed. The reduction factors of > 4 log10 within 5 min for Gram-positive microorganisms claimed for the product using a standard test procedure (ASTM D7907) could not be confirmed in a realistic experimental test set-up even after 10 min of light exposure. The effectiveness against Gram-positive microorganisms should be further investigated under realistic (dry) conditions, including patient care. At this stage, the use of supposedly antimicrobial gloves should not be recommended, as the belief in their efficacy may encourage the misuse of gloves.

Admission to a room previously occupied by patients carrying environmentally robust pathogens implies an increased risk of acquiring those pathogens. Therefore, 'No-touch' automated room disinfection systems, including devices based on UV-C irradiation, are discussed to improve terminal cleaning. It is still unclear if clinical isolates of relevant pathogens behave differently under UV-C irradiation compared to laboratory strains used in the approval process of disinfection procedures. In this study we analysed the susceptibility of well characterized clonally divergent vancomycin-resistant enterococci (VRE) strains, including a linezolid-resistant isolate, against UV-C radiation.Susceptibility against UV-C of ten clonally divergent clinical isolates of VRE was determined in comparison to the commonly used test organism Enterococcus hirae ATCC 10541. Ceramic tiles contaminated with 105 to 106 colony forming units/25 cm² of the different enterococci were positioned at a distance of 1.0 and 1.5 m and irradiated for 20 s, resulting in a UV-C dose of 50 and 22 mJ/cm², respectively. Reduction factors were calculated after quantitative culture of the bacteria recovered from treated and untreated surfaces.Susceptibility to UV-C varied considerably among the strains studied, with the mean value of the most robust strain being up to a power of ten lower compared to the most sensitive strain at both UV-C doses. The two most tolerant strains belonged to MLST sequence types ST80 and ST1283. The susceptibility of the laboratory strain E. hirae ATCC 10541 ranged between the most sensitive and most tolerant isolates for both irradiation doses. However, for UV-C dose of 22 mJ/cm², the reduction of the most tolerant isolate of ST1283 was statistically significantly lower compared to E. hirae ATCC 10541. The most susceptible strains belonged to the MLST sequence types ST117 and ST203.These results indicate that UV-C doses reported in the literature are sufficient for the reduction of commonly used reference strains of enterococci but could be insufficient for the reduction of tolerant patient VRE-isolates in a hospital setting. Therefore, for future studies, the most tolerant clinical isolates should be used to validate automated UV-C devices or longer exposure times should be expected to ensure efficacy in the real world.

BACKGROUND:Worldwide observations revealed increased frequencies of multi-resistant Enterobacterales and resistance genes in hospital wastewater compared to any other type of wastewater. Despite the description of clonal lineages possibly adapted to hospital wastewater, little is known about long term persistence as well as evolution of these lineages. METHODS:In this study, wastewater isolates of different Enterobacterales species from a tertiary care hospital were investigated with 2.5 years distance. Whole Genome Sequencing (WGS) and resistance gene identification were performed for E. coli, C. freundii, S. marcescens, K. pneumoniae, K. oxytoca, and E. cloacae isolates (n = 59), isolated in 2022 and compared with strains isolated from the same wastewater pipeline in 2019 (n = 240). RESULTS:Individual clonal lineages with highly related isolates could be identified in all species identified more than once in 2022 that appear to persist in the wastewater drainage. A common motif of all persistent clonal lineages was the carriage of mobile genetic elements encoding carbapenemase genes with hints for horizontal gene transfer in persistent clones in this environment observed over the 2.5-year period. Multiple plasmid replicons could be detected in both years. In 2022 isolates blaVIM-1 replaced blaOXA-48 as the most common carbapenemase gene compared to 2019. Interestingly, despite a similar abundance of carbapenemase genes (>80% of all isolates) at both time points genes encoding extended spectrum β-lactamases decreased over time. CONCLUSIONS:This data indicates that hospital wastewater continuously releases genes encoding carbapenemases to the urban wastewater system. The evolution of the resident clones as well as the reasons for the selection advantage in this specific ecological niche needs to be further investigated in the future.

BACKGROUND:Various assay methods have been developed to study antimicrobial activity based on contamination of surfaces with different amounts of liquid bacterial suspensions. Since surfaces with frequent hand contact are typically touched in a dry state in clinical settings, these tests may be inappropriate at assessing effectiveness to reduce pathogen transmission. AIM:To investigate a surface previously confirmed to display antimicrobial activity even after drying of small volumes of bacterial suspension (Egger antimicrobial surfaces: EAS) under conditions modelling dry contamination using a touch-transfer method. METHODS:EAS, an antimicrobial copper alloy, as well as a negative control were examined to assess interlaboratory test reproducibility. FINDINGS:Significantly fewer bacteria on EAS after touch transfer and some differences in the touch transmission were detected between the two laboratories. However, an identical assessment of effectiveness for EAS came from both laboratories. Interestingly, despite previously detected antimicrobial efficacy of EAS and the antimicrobial copper alloy after liquid contamination, insufficient activity was observed under dry conditions during a contact time of 4 h by both laboratories. Experiments under standardized air humidity in one laboratory revealed at least for copper a strong influence of humidity on antimicrobial activity. These data indicate that procedures involving contamination of surfaces with organisms suspended in liquids are not directly comparable to dry contamination. CONCLUSION:Since, in the real world of a hospital, organisms are typically transferred between dry surfaces, further standardization of the touch-transfer method is worthwhile for a better understanding of the efficacy of such surfaces.

Regulations for measures to protect against SARS-CoV-2 transmission vary widely around the world, with very strict regulations in Germany where respirators (filtering face piece FFP2 or comparable) are often mandatory. The efficiency of respirators, however, depends essentially on the tight facial fit avoiding the bypass of contaminated air via gaps between mask and wearer's face. The facial fit can be verified in a fit test. The aim of this review was to describe the quantitative fit test results depending on the respirator designs. A literature search revealed 29 suitable studies. Of all respirators with circumferential head straps, three-panel folded dome-shaped respirators showed the best fit (80.8% of 4625 fit tests passed), followed by rigid-dome-shaped respirators (72.4% of 8234 fit tests passed), duckbill-shaped respirators (31.6% of 2120 fit tests passed), and coffee-filter-shaped respirators (30.9% of 3392 fit tests passed). Respirators with ear loops showed very poor tight fit (3.6% of 222 fit tests passed). In four randomized control trials, single-use respirators were not shown to be superior to surgical masks for the prevention of laboratory-confirmed viral respiratory infections, even when adjusted with a fit test. Therefore, we consider the mandatory use of respirators to be disproportionate and not supported by evidence. Further evidence should be generated, in which scenarios respirators might provide an effective benefit as part of occupational health and safety. For situations with confirmed benefits, only high-quality disposable respirators with head straps or respiratory protective equipment of higher protective levels should be used.

Pigmentation, catalase activity and biofilm formation are virulence factors that cause resistance of Staphylococcus aureus to environmental stress factors including disinfectants. In recent years, automatic UV-C room disinfection gained greater importance in enhanced disinfection procedures to improve disinfection success in hospitals. In this study, we evaluated the effect of naturally occurring variations in the expression of virulence factors in clinical S. aureus isolates on tolerance against UV-C radiation. Quantification of staphyloxanthin expression, catalase activity and biofilm formation for nine genetically different clinical S. aureus isolates as well as reference strain S. aureus ATCC 6538 were performed using methanol extraction, a visual approach assay and a biofilm assay, respectively. Log(10) reduction values (LRV) were determined after irradiation of artificially contaminated ceramic tiles with 50 and 22 mJ/cm(2) UV-C using a commercial UV-C disinfection robot. A wide variety of virulence factor expression was observed, indicating differential regulation of global regulatory networks. However, no direct correlation with the strength of expression with UV-C tolerance was observed for either staphyloxanthin expression, catalase activity or biofilm formation. All isolates were effectively reduced with LRVs of 4.75 to 5.94. UV-C disinfection seems therefore effective against a wide spectrum of S. aureus strains independent of occurring variations in the expression of the investigated virulence factors. Due to only minor differences, the results of frequently used reference strains seem to be representative also for clinical isolates in S. aureus.

Background: The efficacy of ultraviolet C (UV -C) radiation against a broad spectrum of micro-organisms has been demonstrated in several studies, but differences in the specific doses and the extent of microbial reduction were found. Furthermore, the conditions of laboratory tests differ greatly from reality, such that efficacy achieved in tests may not necessarily be assumed in reality. Consequently, it is important to investigate the effectiveness of UV-C in representative field trials. The aim was therefore to develop and establish a field test to evaluate automatic UV-C in comparison to manual disinfection.Methods: Before and after disinfection, samples were repeatedly collected from naturally highly contaminated surfaces using the swab technique to obtain representative data sets for disinfected and non-disinfected surfaces. Subsequently, the log reduction values (LRV) and the disinfection success were evaluated for UV-C radiation and full compliant manual disinfection using alcohol-based wipes. Results: Surfaces that are naturally contaminated with bacteria on a regular and nearly uniform basis have been identified as particularly suitable for field testing. Mean contamination was reduced from 23.3 to 1.98 cfu/cm2 (LRV 0.9) and 29.7 to 0.26 cfu/cm2 (LRV 1.2) for UV-C and manual disinfection, respectively. UV-C disinfection achieved 75.5% successful disinfected surfaces, whereas manual disinfection showed 98.1%.Conclusions: Full compliant manual disinfection showed slightly higher LRVs and disinfection success than automatic UV-C disinfection. Successful, operator-independent UV-C disinfection still has the potential to improve disinfection performance in addition to manual disinfection.& COPY; 2023 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

SARS-CoV-2 RNA is frequently identified in patient rooms and it was speculated that the viral load quantified by PCR might correlate with infectivity of surfaces. To evaluate Ct values for the prediction of infectivity, we investigated contaminated surfaces and Ct-value changes after disinfection. Viral RNA was detected on 37 of 143 investigated surfaces of an ICU. However, virus isolation failed for surfaces with a high viral RNA load. Also, SARS-CoV-2 could not be cultivated from surfaces artificially contaminated with patient specimens. In order to evaluate the significance of Ct values more precisely, we used surrogate enveloped bacteriophage Φ6. A strong reduction in Φ6 was achieved by three different disinfection methods. Despite a strong reduction in viability almost no change in the Ct values was observed for UV-C and alcoholic surface disinfectant. Disinfection using ozone resulted in a lack of Φ6 recovery as well as a detectable shift in Ct values indicating strong degradation of the viral RNA. The observed lack of significant effects on the detectable viral RNA after effective disinfection suggest that quantitative PCR is not suitable for predicting the infectivity of SARS-CoV-2 on inanimate surfaces. Ct values should therefore not be considered as markers for infectivity in this context.

The potential impact of music-making on air quality around musicians was inferred at the outset of the SARS-CoV-2 pandemic from measurements on individual musical instruments and from theoretical considerations. However, it is unclear to what extent playing together in an orchestra under optimal ventilation conditions really increases infection risks for individual musicians. In this study, changes in indoor air quality were assessed by measuring common parameters, i.e., temperature, relative humidity, and carbon dioxide, along with particle counting and determining the presence of airborne pharyngeal bacteria under different seating arrangements. The study was conducted in cooperation with a professional orchestra on a stage ventilated by high volume displacement ventilation. Even with a full line-up, the particle load was only slightly influenced by the presence of the musicians on stage. At the same time, however, a clear increase in pharyngeal flora could be measured in front of individual instrument groups, but independent of seat spacing. Simultaneous measurement of various air parameters and, above all, the determination of relevant indicator bacteria in the air, enables site-specific risk assessment and safe music-making even during a pandemic.

Hospitals are one of the main reservoirs of multi-resistant Enterobacterales (MRE). As MRE are resistant to the most frequently used antibiotics, therapy for patients with MRE infections is challenging. It has been previously described that MRE from hospital wastewater can pass into municipal wastewater and even surface water. In this study, we investigated the diversity and epidemiology of MRE in the wastewater of a large tertiary care hospital. Wastewater samples were collected for a four-day period and tested for the presence of Enterobacterales resistant to 3rd gen. cephalosporins. Representative isolates were further characterized by whole genome sequencing. In 120 β-glucuronidase-producing isolates, 68 Escherichia coli and, interestingly, also 52 Citrobacter freundii were identified. In 120 β-glucosidase-producing isolates 45 Serratia marcescens, 34 Klebsiella oxytoca, 32 Enterobacter cloacae and 9 Klebsiella pneumoniae were observed. For all species various MLST sequence types and different clusters of resistance genes were determined, showing a great diversity within the different Enterobacterales, further corroborated by clonal analysis performed by cgMLST. The most prominent clone was wastewater associated E. coli ST635, which accounted for 47.1% of all E. coli isolates. Interestingly, 45.6% of E. coli, 88.5% of C. freundii, 95.6% of S. marcescens, 91.2% of K. oxytoca, 96.9% of E. cloacae and 88.9% of K. pneumoniae isolates carried a carbapenemase gene, indicating a high burden with carbapenemase-producing Enterobacterales. Comparison with clinical isolates from the same hospital displayed few clonal matches. One wastewater isolate of K. pneumoniae was identified to be closely related compared to a clone that had been introduced into the hospital during an outbreak four years earlier. One E. coli isolate was identified as identical to an isolate from a patient, with inpatient stay during the sampling period. The data obtained in this study highlight the problem of antibiotic resistance of Enterobacterales in hospital wastewater. In particular, the clustered occurrence of carbapenemase genes is of great concern and underscores the problem of increasingly scarce antibiotic options against these bacteria.

The extent of monkeypox virus environmental contamination of surfaces is unclear. We examined surfaces in rooms occupied by two monkeypox patients on their fourth hospitalisation day. Contamination with up to 10(5)viral copies/cm(2) on inanimate surfaces was estimated by PCR and the virus was successfully isolated from surfaces with more than 106 copies. These data highlight the importance of strict adherence of hospital staff to recommended protective measures. If appropriate, pre-exposure or early post-exposure vaccination should be considered for individuals at risk.

BACKGROUND:The presence of coronaviruses on surfaces in the patient environment is a potential source of indirect transmission. Manual cleaning and disinfection measures do not always achieve sufficient removal of surface contamination. This increases the importance of automated solutions in the context of final disinfection of rooms in the hospital setting. Ozone is a highly effective disinfectant which, combined with high humidity, is an effective agent against respiratory viruses. Current devices allow continuous nebulization for high room humidity as well as ozone production without any consumables. AIM:In the following study, the effectiveness of a fully automatic room decontamination system based on ozone was tested against bacteriophage Φ6 (phi 6) and bovine coronavirus L9, as surrogate viruses for the pandemic coronavirus SARS-CoV-2. METHODS:For this purpose, various surfaces (ceramic tile, stainless steel surface and furniture board) were soiled with the surrogate viruses and placed at two different levels in a gas-tight test room. After using the automatic decontamination device according to the manufacturer's instructions, the surrogate viruses were recovered from the surfaces and examined by quantitative cultures. Then, reduction factors were calculated. FINDINGS:The ozone-based room decontamination device achieved virucidal efficacy (reduction factor >4 log10) against both surrogate organisms regardless of the different surfaces and positions confirming a high activity under the used conditions. CONCLUSION:Ozone is highly active against SARS-CoV-2 surrogate organisms. Further investigations are necessary for a safe application and efficacy in practice as well as integration into routine processes.

Abstract Background Acinetobacter baumannii is an emerging multiresistant Gram-negative rod, which has caused multiple hospital outbreaks. A. baumannii can display a high ability to survive on inanimate surfaces. Therefore, cleaning and disinfection is an important part in the prevention of A. baumannii transmission. No-touch room decontamination is performed with increasing frequency to reach more standardization in hospital cleaning. In this study, we investigated the efficacy of an automated room decontamination using ozone (Sterisafe™ pro) against A. baumannii under complex room conditions. Methods A Carbapenemase-producing A. baumannii outbreak strain was analyzed with respect to its ability to survive on dry surfaces. The Sterisafe™ pro instrument was used in a patient room with an attached bathroom. The A. baumannii strain was dried on three different carriers (ceramic tiles, stainless steel, solid core furniture board) and placed at eight different positions in the rooms. A standard disinfection cycle (80 ppm ozone; 90 % RH; 60 minutes) was conducted in three independent experiments. Results The A. baumannii strain displayed a long-term survival on surfaces under dry conditions sufficient for further disinfection experiments. Interestingly, the mean reduction rates of A. baumannii dried on three surfaces displayed significant differences. Reduction rates greater than 5 log were reached on all stainless steel and ceramic carriers even under the complex room conditions using the standard disinfection cycle of the Sterisafe™ pro instrument. In contrast, on furniture board individual carriers displayed reduction rates of even less than 4 log. The mean reduction rate was still 5 log for A. baumannii on furniture board. Conclusion A. baumannii dried on different surfaces display a differential susceptibility against automated ozone disinfection. However, the SterisafeTM pro instrument displayed a sufficient reduction of A. baumannii for all tested surfaces even under complex room conditions. The individual behavior of A. baumannii on different materials indicate the necessity for the validation of automated room decontamination systems under varying conditions. Disclosures All authors: No reported disclosures.