Virucidal activity of oral, hand, and surface disinfectants against respiratory syncytial virus.

BACKGROUND: Respiratory syncytial virus (RSV) is known as a major cause of respiratory tract infection in adults and children. Human-to-human transmission occurs via droplets as well as direct and indirect contact (e.g. contaminated surfaces or hands of medical staff). Therefore, applicable hygiene measures and knowledge about viral inactivation are of utmost importance. AIM: To elucidate the disinfection profile of RSV. METHODS: The study evaluated the virucidal efficacy of oral rinses specifically designed for children, World Health Organization (WHO)-recommended hand-rub formulations, and ethanol, as well as 2-propanol against RSV in a quantitative suspension test (EN14476). The stability of RSV on stainless steel discs was assessed and its inactivation by different surface disinfectants (EN16777) investigated. FINDINGS: All tested oral rinses except one reduced infectious viral titres to the lower limit of quantification. The two WHO-recommended hand-rub formulations as well as 30% ethanol and 2-propanol completely abolished the detection of infectious virus. Infectious RSV was recovered after several days on stainless steel discs. However, RSV was efficiently inactivated by all tested surface disinfectants based on alcohol, aldehyde, or hydrogen peroxide. CONCLUSION: Oral rinses, all tested hand-rub formulations as well as surface inactivation reagents were sufficient for RSV inactivation in vitro.

Evaluation of temperature, drying time and other determinants for the recovery of Gram-negative bacterial pathogens in disinfectant efficacy testing.

BACKGROUND: In the clinical setting, surface disinfection is an important measure to reduce the risk of cross transmission of micro-organisms and the risk of nosocomial infections. Standardized methods can be used to evaluate disinfection procedures, as well as the effectiveness of the active ingredients used for disinfection. However, despite standardization, the results of such methodologies are still determined by several factors, and incorrect results may lead to invalid assumptions about the effectiveness of a disinfectant, posing significant health risks for patients and health personnel. AIM: The objective of this study was to evaluate several determinants for the recovery of Pseudomonas aeruginosa and other test organisms to establish their influence on the results of standardized disinfection methodologies, and to find Gram-negative strains that can be used as suitable replacements for P. aeruginosa. METHODS: The effects of inoculum application method, drying time, temperature and carrier material on the survival and recovery of the test organisms were evaluated using Student's t-test, one-way analysis of variance and Tukey's multiple comparison test. FINDINGS AND CONCLUSIONS: Temperature, drying time, application method and carrier material were found to affect the recovery of P. aeruginosa cells significantly, and therefore influence the outcome of the methodologies used. This study also showed thatP. aeruginosa could be replaced with the Gram-negative species Acinetobacter baumannii, a test organism used in many standardized methodologies, which responds better under the same circumstances and has a behaviour similar to that of P. aeruginosa in disinfectant efficacy tests.

Survival and inactivation of hepatitis E virus on inanimate surfaces.

BACKGROUND: Hepatitis E virus (HEV) is the most common cause of acute viral hepatitis, and mainly transmitted via faecal-oral contamination or consumption of contaminated food products. However, limited data on the surface stability and HEV sensitivity to chemical disinfectants are available. AIM: To establish an HEV-based carrier assay to evaluate its surface stability and the virucidal activity of nine surface disinfectants. METHODS: A recently developed robust HEV-3 cell culture system for an HEV-based carrier assay. FINDINGS: Alcohol-based disinfectants were insufficient to eliminate HEV infectivity, whereas disinfectants based on aldehyde, peracetic acid, oxygen, and/or quaternary ammonium inactivated HEV. CONCLUSION: These findings have strong implications for the recommendation of evidence-based hygiene guidelines to reduce HEV transmission.

Evaluation of the substitution of poliomyelitis virus for testing virucidal activities of instrument and surface disinfection.

The Global Polio Eradication initiative has the goal to eradicate poliomyelitis worldwide. This means that poliomyelitisvirus type 1 strain LSc 2ab (PV-1) can no longer be used for the evaluation of virucidal activity of chemical disinfectants. This study evaluated murine parvovirus ATCC VR 1346 (minute virus of mice) as suitable surrogate for PV-1 when testing virucidal activity of biocides in instrument and surface disinfectants. Suspension testing in different laboratories with two commercially available active biocidal substances based on glutaraldehyde (0.01-0.25%) and peracetic acid (0.005-0.1%) with an exposure time of 30 min was performed. Both pathogens showed comparable susceptibility and dose-dependent reduction of virus titres following German and European Guidelines.

[Microbiological diagnosis of implant-associated infections : Retrospective analysis of 133 patients in an arthroplasty center]. / Mikrobiologische Diagnostik bei implantatassoziierten Infektionen : Retrospektive Analyse von 133 Patienten in einem Endoprothetikzentrum.

BACKGROUND: Because standardized microbiological cultures of puncture fluids and tissue samples often do not provide pathogen detection in implant-associated infections, sonication and polymerase chain reaction (PCR) are used additionally today. OBJECTIVES: Pathogen spectra and previous microbiological standards are examined for agreement of results using the new methods sonication and PCR. MATERIALS AND METHODS: In this descriptive, retrospective observational study, we evaluated the data of 133 patients in whom a joint prosthesis, osteosynthesis material or a spacer was removed during revision surgery with suspected implant-associated infection and sent for sonication. RESULTS: Pathogen detection was achieved by culture of peri-implant material in 40.1% and by sonication in 42.5%. In each case, coagulase-negative staphylococci were detected most frequently. Overall, the results were consistent in 71.7% of cases. In the discrepant cases, more anaerobes could be detected by sonication, especially for osteosynthesis material and knee prostheses. PCR analyses in 21 cases showed pathogen detection in 14.3% and agreement with the results of peri-implant tissue culture and sonication in 57.1% and 66.7%, respectively. CONCLUSIONS: The present results indicate a gain in sensitivity of sonication, especially for anaerobes that are difficult to grow, and a gain in specificity through sonication. PCR analyses should be reserved for specific questions.

Virucidal activity of nasal sprays against severe acute respiratory syndrome coronavirus-2.

The highest viral loads of severe acute respiratory syndrome coronavirus-2 are detectable in the oral cavity, so a potential reduction of infectious virus by nasal and oral sprays could reduce transmission. Therefore, the inactivation capacity of nine nasal and oral sprays was evaluated according to EN 14476. One nasal spray based on sodium hypochlorite and one oral spray containing essential oils reduced viral titres by two to three orders of magnitude. Although clinical data are still sparse, nasal and oral sprays display a more convenient application for elderly people or those who are unable to rinse/gargle.

Virucidal efficacy of an ozone-generating system for automated room disinfection.

Besides conventional prevention measures, no-touch technologies based on gaseous systems have been introduced in hospital hygiene for room disinfection. The whole-room disinfectant device Sterisafe Pro, which creates ozone as a biocidal agent, was tested for its virucidal efficacy based on Association Française de Normalisation Standard NF T 72-281:2014. All test virus titres were reduced after 150 and 300 min of decontamination, with mean reduction factors ranging from 2.63 (murine norovirus) to 3.94 (simian virus 40). These results will help to establish realistic conditions for virus inactivation, and assessment of the efficacy of ozone technology against non-enveloped and enveloped viruses.

[Long-term care facilities during the COVID-19 pandemic : Considerations on the way back to normality]. / Langzeitpflegeeinrichtungen in der COVID-19-Pandemie : Überlegungen auf dem Weg zurück in die Normalität.

Long-term care facilities (LTCF) were and are particularly affected by the COVID-19 pandemic. The dimensions of the outbreaks and the high mortality among residents led to massive restrictions in LTCFs, especially in the area of social contacts and activities but also in areas of medical care. With the start of vaccinations and the improved testing options, the situation has now changed and existing restrictions must be evaluated to determine whether they are still appropriate. In an interprofessional and interdisciplinary group of experts, considerations have been formulated on how a way back to normality could look like in LTCFs.

Improved method for tuberculocidal and mycobactericidal activity testing of disinfectants based on the European Standard EN 14348.

Safe measurements to prevent the transmission of (multidrug-resistant) mycobacteria such as disinfection are essential in healthcare settings. In Europe chemical disinfectants are tested for their tuberculocidal and mycobactericidal efficacy by the internationally accepted test procedure described in EN 14348. However, especially for amine-based disinfectants, invalid results may occur by this procedure due to insufficient neutralization. In this multi-laboratory study the procedure described in EN 14348 was optimized by a combination of chemical neutralization and membrane filtration in order to obtain a valid and secure method especially for amine-based disinfectants.

Potential sources, modes of transmission and effectiveness of prevention measures against SARS-CoV-2.

During the current SARS-CoV-2 pandemic new studies are emerging daily providing novel information about sources, transmission risks and possible prevention measures. In this review, we aimed to comprehensively summarize the current evidence on possible sources for SARS-CoV-2, including evaluation of transmission risks and effectiveness of applied prevention measures. Next to symptomatic patients, asymptomatic or pre-symptomatic carriers are a possible source with respiratory secretions as the most likely cause for viral transmission. Air and inanimate surfaces may be sources; however, viral RNA has been inconsistently detected. Similarly, even though SARS-CoV-2 RNA has been detected on or in personal protective equipment (PPE), blood, urine, eyes, the gastrointestinal tract and pets, these sources are currently thought to play a negligible role for transmission. Finally, various prevention measures such as handwashing, hand disinfection, face masks, gloves, surface disinfection or physical distancing for the healthcare setting and in public are analysed for their expected protective effect.

Efficacies of the original and modified World Health Organization-recommended hand-rub formulations.

The World Health Organization (WHO) hand-rub formulations have been in use around the world for at least the past 10 years. The advent of coronavirus disease 2019 (COVID-19) has further enhanced their use. We reviewed published efficacy data for the original and modified formulations. Only efficacy data according to the European Norms (EN) were found. The bactericidal efficacy of the original formulations was, under practical conditions, partly insufficient (EN 1500, only effective in 60 s; EN 12791, efficacy too low in 5 min). The first modification with higher alcohol concentrations improves their efficacy as hygienic hand rub (effective in 30 s). The second (0.725% glycerol) and third (0.5% glycerol) modification improves their efficacy for surgical hand preparation (effective in 5 and 3 min). The original and second modified formulations were tested and demonstrate activity against enveloped viruses including severe acute resiratory syndrome coronavirus 2 (SARS-CoV-2) in 30 s. The ethanol-based formulation is also active against some non-enveloped test viruses in 60 s (suspension tests, EN 14476). In-vivo data on the formulations would provide a more reliable result on the virucidal efficacy on contaminated hands but are currently not available. Nevertheless, the most recent modifications should be adopted for use in healthcare.

Voriconazole plus terbinafine combination antifungal therapy for invasive Lomentospora prolificans infections: analysis of 41 patients from the FungiScope® registry 2008-2019.

OBJECTIVES: Lomentospora prolificans is an emerging cause of serious invasive fungal infections. Optimal treatment of these infections is unknown, although voriconazole-containing treatment regimens are considered the treatment of choice. The objective of this study was to evaluate the role of combination antifungal therapy for L. prolificans infections. METHODS: We performed a retrospective review of medical records of patients with invasive L. prolificans infection diagnosed between 1 January 2008 and 9 September 2019 that were documented in the FungiScope® registry of rare invasive fungal infections. We compared clinical outcomes between antifungal treatment strategies. RESULTS: Over the study period, 41 individuals with invasive L. prolificans infection from eight different countries were documented in the FungiScope® registry. Overall, 17/40 (43%) had treatment response/stable disease and 21/40 (53%) had a fatal outcome attributed to invasive fungal infection. Combination antifungal therapy was associated with increased 28-day survival (15/24 survived versus 4/16 receiving monotherapy; p 0.027) and the combination voriconazole plus terbinafine trended to be associated with higher rates of treatment success (10/16, 63%, 95% CI 35%-85%) compared with other antifungal treatment regimens (7/24, 29%, 95% CI 13%-51%, p 0.053). In Kaplan-Meier survival analysis there was a higher survival probability in individuals receiving the voriconazole/terbinafine combination compared with other antifungal regimens (median survival 150 days versus 17 days). CONCLUSIONS: While overall mortality was high, combination antifungal treatment, and in particular combination therapy with voriconazole plus terbinafine may be associated with improved treatment outcomes compared with other antifungal regimens for the treatment of invasive L. prolificans infections.

Performance of the AsperGenius® PCR assay for detecting azole resistant Aspergillus fumigatus in BAL fluids from allogeneic HSCT recipients: A prospective cohort study from Essen, West Germany.

In this study a commercially available multiplex real-time PCR (AsperGenius®) was evaluated for its efficacy in detecting Aspergillus fumigatus and azole resistance markers in comparison with conventional culture methods and galactomannan (GM) testing from BAL fluids in allogeneic HSCT recipients. Between January 2015 and May 2017 100 allogeneic HSCT recipients with pulmonary infiltrates and suspicion of invasive fungal infection were recruited to the study from a tertiary care center in Germany. BAL fluid was routinely assessed using the following diagnostic tests: AsperGenius® PCR assay, GM testing (cut-off: 1.0) and conventional culture. Susceptibility testing of azoles was performed by using Etest and, in case presenting elevated MICs, PCR for mutations in the cyp51A gene was carried out. Criteria of EORTC/MSG were used to classify the patients for invasive fungal disease. According to the EORTC/MSG criteria 23 patients presented with probable invasive aspergillosis (IA). Aspergillus PCR showed a sensitivity of 65% for probable IA cases. A combination of PCR and GM results in BAL displayed a sensitivity of 96% (22/23) and 100% specificity. Mutations in the cyp51A gene were detected by PCR in three cases (3/23; 13%) which were also found resistant with the culture method. In one case a Y121F/T289A mutation and in two cases a L98H were found. The combination of a commercial Aspergillus PCR assay and GM testing from BAL demonstrated a high sensitivity and specificity for diagnosing IA in allogeneic HSCT recipients. The Aspergillus PCR assay was not superior in detecting azole resistant A. fumigatus compared to culture.

In-vitro activity of active ingredients of disinfectants against drug-resistant fungi.

The biocidal activities of peracetic acid and ethanol were tested against nine clinical fungal isolates and four reference strains. Ethanol was active (≥4.0 log10 reduction) against yeasts at a concentration of 50% v/v and against moulds at 80% v/v. Exposure times in both cases were 1 min. Peracetic acid was active as a 0.25% solution against yeasts and as a 0.5% solution against moulds; exposure times in both cases were 5 min. Compared with the reference strains, clinical isolates, including multi-drug-resistant strains, showed similar or higher sensitivity to the active ingredients of disinfectants in vitro.

Detection of invasive pulmonary aspergillosis in critically ill patients by combined use of conventional culture, galactomannan, 1-3-beta-D-glucan and Aspergillus specific nested polymerase chain reaction in a prospective pilot study.

Invasive pulmonary aspergillosis (IPA) is an emerging and life-threatening infectious disease in patients admitted to the intensive care unit (ICU). Most diagnostic studies are conducted in hematological patients and results cannot readily be transferred to ICU patients lacking classical host factors. In a multicenter, prospective clinical trial including 44 ICU patients, hematological (n = 14) and non-hematological patients (n = 30), concurrent serum and bronchoalveolar lavage (BAL) samples were analyzed by conventional culture, galactomannan (GM), 1-3-beta-D-glucan (BDG) as well as an Aspergillus specific nested polymerase chain reaction (PCR). Nine patients (20%) had putative IPA according to AspICU classification. GM and PCR showed superior performance in BAL with sensitivity/specificity of 56%/94% and 44%/94% compared to 33%/97% and 11%/94% in serum. Despite better sensitivity of 89%, BDG showed poor specificity of only 31% (BAL) and 26% (serum). Combination of GM and PCR (BAL) with BDG (serum) resulted in 100% sensitivity, but also reduced specificity to 23%. Whereas mean GM levels were significantly higher in hematological patients BDG and PCR did not differ between hematological and non-hematological patients. Under present clinical conditions test combinations integrating both BAL and blood samples are advantageous. BDG might best serve as possible indicator for ruling out IPA. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT01695499. First posted: September 28, 2012, last update posted: May 8, 2017.

Airborne Aspergillus fumigatus spore concentration during demolition of a building on a hospital site, and patient risk determination for invasive aspergillosis including azole resistance.

BACKGROUND: Invasive aspergillosis (IA) in immunocompromised patients has been associated with demolition in or adjacent to hospitals. In recent years, azole-resistant clinical isolates of Aspergillus fumigatus, the most common agent of IA, have emerged in Western Europe and are spreading globally. AIM: To determine the potential risk of IA, including azole resistance, in patients caused by demolition of a hospital building. METHODS: Air sampling before, during and after demolition, screening for azole resistance, genotyping of non-susceptible isolates, and comparing those with strains from patients with azole-resistant IA during demolition. FINDINGS: Mean concentrations of A. fumigatus spores did not differ significantly between the three periods before [17.5 colony-forming units (cfu)/m³], during (20.8 cfu/m³) (P=0.26) and after (17.7 cfu/m³) demolition (P=0.33). No significant difference in IA cases documented by clinicians was found when comparing the timeframe of demolition with the previous year (44 vs 42 cases). Thirty of 200 A. fumigatus isolates (15%) showed azole resistance. Genotyping by microsatellite polymerase chain reaction of the azole-resistant environmental and clinical isolates showed a polyclonal distribution. CONCLUSIONS: The results suggest that with implemented preventive measures, there is no increased risk for IA, including azole resistance, in immunocompromised patients during outdoor demolition work. Further prospective studies are needed to confirm these findings.

Prevalence and characterization of azole-resistant Aspergillus fumigatus in patients with cystic fibrosis: a prospective multicentre study in Germany.

Objectives: Aspergillus fumigatus is the most prevalent filamentous fungus in the respiratory tract of patients with cystic fibrosis (CF). The aim of this prospective multicentre study was to investigate the prevalence of azole-resistant A. fumigatus (ARAF) in respiratory secretions from CF patients across Germany and to characterize ARAF isolates by phenotypic and molecular methods. Methods: Twelve tertiary care centres from Germany participated in the study. In total, 2888 A. fumigatus isolates from 961 CF patients were screened for ARAF by using azole-containing agar plates. Antifungal susceptibility testing of isolates was performed by broth microdilution according to EUCAST guidelines. Analysis of mutations mediating resistance was performed using PCR and sequencing of the cyp51A gene. Furthermore, genotyping by microsatellite PCR was performed. Results: Of a total of 2888 A. fumigatus isolates, 101 isolates from 51 CF patients were found to be azole resistant (prevalence per patient 5.3%). The Essen centre had the highest prevalence (9.1%) followed by Munich (7.8%), Münster (6.0%) and Hannover (5.2%). Most ARAF isolates (n = 89) carried the TR34/L98H mutation followed by eight G54E/R, one TR46/Y121F/T289A and one F219S mutation. In two isolates no mutation was found. Genotyping results showed no major clustering. Forty-five percent of CF patients with ARAF had previously received azole therapy. Conclusions: This is the first multicentre study analysing the prevalence of ARAF isolates in German CF patients. Because of a resistance rate of up to 9%, susceptibility testing of A. fumigatus isolates from CF patients receiving antifungal treatment should be part of standard diagnostic work-up.

Azole Resistance in Aspergillus fumigatus in Patients with Cystic Fibrosis: A Matter of Concern?

Aspergillus fumigatus is the most frequent filamentous fungus isolated from respiratory specimens from patients with cystic fibrosis (CF). Triazoles are the most widely used antifungals in the treatment of allergic bronchopulmonary aspergillosis (ABPA) and invasive aspergillosis (IA) in CF patients. Treatment success could be severely compromised by the occurrence of azole-resistant A. fumigatus (ARAf), which is increasingly reported worldwide from both clinical samples and the environment. In previous studies, ARAf has been detected in up to 8% of CF patients. Isolates from CF patients requiring antifungal treatment should therefore be routinely subjected to antifungal susceptibility testing. The optimal treatment of ABPA or IA in CF patients with azole-resistant isolates has not been established; treatment options include liposomal amphotericin B i.v. and/or echinocandins i.v.

The lung microbiome in patients with pneumocystosis.

BACKROUND: Pneumocystis jirovecii pneumonia (PCP) is an opportunistic fungal infection that is associated with a high morbidity and mortality in immunocompromised individuals. In this study, we analysed the microbiome of the lower respiratory tract from critically ill intensive care unit patients with and without pneumocystosis. METHODS: Broncho-alveolar fluids from 65 intubated and mechanically ventilated intensive care unit patients (34 PCP+ and 31 PCP- patients) were collected. Sequence analysis of bacterial 16S rRNA gene V3/V4 regions was performed to study the composition of the respiratory microbiome using the Illumina MiSeq platform. RESULTS: Differences in the microbial composition detected between PCP+ and PCP- patients were not statistically significant on class, order, family and genus level. In addition, alpha and beta diversity metrics did not reveal significant differences between PCP+ and PCP- patients. The composition of the lung microbiota was highly variable between PCP+ patients and comparable in its variety with the microbiota composition of the heterogeneous collective of PCP- patients. CONCLUSIONS: The lower respiratory tract microbiome in patients with pneumocystosis does not appear to be determined by a specific microbial composition or to be dominated by a single bacterial species.