Dental aerosol-producing treatments: Comparison of contamination patterns of face shields and surgical masks.

During the COVID-19 pandemic, dental face shields were recommended to protect the eyes. This study aimed to examine to what extent face shield and mask contamination differ when a pre-procedural mouth rinsing with Chlorhexidine (CHX) is conducted before treatment. In this prospective, randomized study, three groups of subjects were formed (rinsing with 0.1% CHX, water, or no rinsing (control) before aerosol-producing treatments). After each of the 301 treatments, the practitioner's face shield was swabbed with eSwab and the mask was brought into contact with agar plates. Sampling was done from the exterior surface only. Samples were cultured for 48 h at 35 °C under aerobic and anaerobic conditions. Bacteria were classified by phenotypic characteristics, biochemical test methods, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Colony-forming units were counted and mean values were compared (WSR, H-test, U-test, p < 0.05). Within each subject group, face shields showed significantly more contamination than surgical masks (control group: 350 CFU, 50 CFU; intervention water: 270 CFU, 40 CFU; intervention CHX: 250 CFU, 30 CFU). Comparison of face shields of the different subject groups did not reveal any statistically significant differences. However, CHX resulted in a statistically significant bacterial reduction on surgical masks compared to the water and control group (control: 50 CFU, intervention water: 40 CFU, intervention CHX: 30 CFU). Contamination of face shields and surgical masks was highest in the control group, followed by the water group, and lowest in the intervention group with CHX. Streptococcus spp. and Staphylococcus spp. dominated, representing the oral and cutaneous flora. Contamination of masks worn with or without face shields did not differ. Presumably, face shields intercept first splashes and droplets, while the masks were mainly exposed to bioaerosol mist. Consequently, face shields protect the facial region and surroundings from splashes and droplets, but not the mask itself. A pre-procedural mouth rinse with CHX had no statistically significant reducing effect on contamination of the face shield, but a statistically significant reducing effect was observed on contamination of the mask.

Detection of viable oral bacteria of the patient on the surgical mask of dentists.

INTRODUCTION AND AIM: Bioaerosols contaminate the personal protective equipment (PPE), especially masks. The PPE harbors microorganisms from various sources. However, no previous studies have investigated the specific sources of bacteria found on used masks and their correlation with those from the treated patient. SETTING, DESIGN, MATERIAL AND METHODS: Intraoral samples from the patient were collected prior to dental aerosol-producing treatments using a nylon flock fiber swab. After treatment, the practitioner's mask was imprinted onto agar plates. MAIN OUTCOME METHODS: Following cultivation, colony forming units were counted and identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). After the samples were analyzed, the intraoral samples as well as the mask samples were assessed for the presence of identical species, which were subsequently quantified. RESULTS: 126 treatments were included. One species match occurred most frequently (26.2%), followed by two (11.9%%) and three or more (3.97%). In the intraoral samples, Neisseria subflava occurred most often, within mask samples Staphylococcus epidermidis were detected most. Staphylococcus aureus could be cultivated three times more often in intraoral samples than on the mask. DISCUSSION AND CONCLUSION: Oral microorganisms originating from the patient's oral cavity can be found on the outside of masks. When using PPE during treatments, it should therefore always be in mind that potentially pathogenic microorganisms may land on the mask becoming a source of for itself.

A multi-pronged approach to improve blood culture diagnostics in different clinical departments: a single-centre experience.

PURPOSE: Blood culture (BC) diagnostics are influenced by many factors. We performed a targeted interdisciplinary analysis to analyse effects of various measures on BC diagnostics performance. METHODS: A diagnostic stewardship initiative was conducted at two intervention and two control wards in a German tertiary level hospital. The initiative comprised staff training on the correct indications and sampling for BC, implementation of information cards, labels to identify the collection site, regular BC bottle feedback including the number of bottles, filling volumes and identified pathogens; and the use of a specific sampling device (BD Vacutainer®). Before and after the interventions, two three-month measurement periods were performed, as well as a one-month follow-up period to assess the sustainability of the conducted measures. RESULTS: In total, 9362 BC bottles from 787 patients were included in the analysis. The number of BCs obtained from peripheral venous puncture could be increased at both intervention wards (44.0 vs. 22.2%, 58.3 vs. 34.4%), while arterial sampling could be reduced (30.6 vs. 4.9%). A total of 134 staff members were fully trained. The intervention led to a considerable increase in BC knowledge (from 62.4 to 79.8% correct answers) with differences between the individual professional groups. Relevant reduced contamination rates could be detected at both intervention wards. CONCLUSIONS: As knowledge on the correct BC sampling and strategies to reduce contamination varies considerably between clinical departments and healthcare professionals, a targeted training should be adapted to the specific needs of the individual professional groups. An additional filling device is not necessary.

The Impact of the Microbiological Vaginal Swab on the Reproductive Outcome in Infertile Women.

BACKGROUND: The thesis on which this paper is based intended to investigate whether the result of the microbiological vaginal swab has an influence on the outcome of the fertility treatment. METHODS: The microbiological vaginal swabs of patients who received fertility treatment at Saarland University Hospital were evaluated. Depending on the microorganisms detected, the swab result was classified as inconspicuous, intermediate, or conspicuous. The SPSS software was used to determine the correlation between the swab result and the outcome of the fertility treatment. RESULTS: Dysbiosis was associated with a worse outcome of fertility treatment. The pregnancy rate with a conspicuous swab was 8.6%, whereas it was 13.4% with an inconspicuous swab. However, this association was not statistically significant. Furthermore, an association of endometriosis with dysbiosis was found. Endometriosis was more frequent with a conspicuous swab result than with an inconspicuous result (21.1% vs. 17.7%), yet the correlation was not statistically significant. However, the absence of lactobacilli was significantly associated with endometriosis (p = 0.021). The association between endometriosis and a lower pregnancy rate was also statistically significant (p = 0.006). CONCLUSION: The microbiological vaginal and cervical swabs can be used as predictors for the success of fertility treatments. Further studies are needed to assess the impact of transforming a dysbiotic flora into a eubiotic environment on the success of fertility treatments.

CHX and a Face Shield Cannot Prevent Contamination of Surgical Masks.

Background: Bacterial contamination on surgical masks puts a threat to medical staff and patients. The aim of the study was to investigate its contamination during dental treatments, wearing a face shield and performing a pre-procedural mouth rinsing with chlorhexidine (CHX). Methods: In this prospective, randomized study, 306 treatments were included, 141 single-tooth (restorations) and 165 total dentition treatments (preventive or periodontal supportive ultrasonic application). A total of three groups (each: n = 102) were formed: participants rinsed for 60 s with 0.1 % CHX or with water before treatment, and, for control, a non-rinsing group was included. In view of the COVID-19 pandemic, a face shield covering the surgical mask enhanced personal protective equipment. After treatment, masks were imprinted on agar plates and incubated at 35°C for 48 h. Bacteria were classified by phenotypic characteristics, biochemical assay methods, and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). Colonies (CFU) were counted and mean values were compared (Kruskal-Wallis-, U test, p < 0.05). Results: Chlorhexidine led to a statistically significant reduction of bacterial contamination of the surgical mask (mean: 24 CFU) in comparison with water (mean: 47 CFU) and non-rinsing (mean: 80 CFU). Furthermore, rinsing with water reduced CFU significantly in comparison with the non-rinsing group. There were no significant differences between single or total dentition treatments. Streptococcus spp., Staphylococcus spp., Micrococcus spp., and Bacillus spp. dominated, representing the oral and cutaneous flora. Conclusion: A pre-procedural mouth rinse is useful to reduce the bacterial load of the surgical mask. However, contamination cannot be prevented completely, even by applying a face shield. In particular, during pandemic, it is important to consider that these additional protective measures are not able to completely avoid the transmission of pathogens bearing aerosols to the facial region. If antiseptic rinsing solutions are not available, rinsing with water is also useful.

German Multicenter Study Analyzing Antimicrobial Activity of Ceftazidime-Avibactam of Clinical Meropenem-Resistant Pseudomonas aeruginosa Isolates Using a Commercially Available Broth Microdilution Assay.

Multidrug resistance is an emerging healthcare issue, especially concerning Pseudomonas aeruginosa. In this multicenter study, P. aeruginosa isolates with resistance against meropenem detected by routine methods were collected and tested for carbapenemase production and susceptibility against ceftazidime-avibactam. Meropenem-resistant isolates of P. aeruginosa from various clinical materials were collected at 11 tertiary care hospitals in Germany from 2017−2019. Minimum inhibitory concentrations (MICs) were determined via microdilution plates (MICRONAUT-S) of ceftazidime-avibactam and meropenem at each center. Detection of the presence of carbapenemases was performed by PCR or immunochromatography. For meropenem-resistant isolates (n = 448), the MIC range of ceftazidime-avibactam was 0.25−128 mg/L, MIC90 was 128 mg/L and MIC50 was 16 mg/L. According to EUCAST clinical breakpoints, 213 of all meropenem-resistant P. aeruginosa isolates were categorized as susceptible (47.5%) to ceftazidime-avibactam. Metallo-ß-lactamases (MBL) could be detected in 122 isolates (27.3%). The MIC range of ceftazidime-avibactam in MBL-positive isolates was 4−128 mg/L, MIC90 was >128 mg/L and MIC50 was 32 mg/L. There was strong variation in the prevalence of MBL-positive isolates among centers. Our in vitro results support ceftazidime-avibactam as a treatment option against infections caused by meropenem-resistant, MBL-negative P. aeruginosa.

Molecular epidemiology and antimicrobial resistance of Clostridioides difficile in Germany, 2014-2019.

Clostridioides difficile is a Gram positive spore-forming rod and mainly responsible for nosocomial diarrhea in developed nations. Molecular and antimicrobial surveillance is important for monitoring the strain composition including genotypes of high epidemiological importance such as ribotype 027 (RT027) and corresponding resistance patterns. 1535 isolates obtained from samples sent between 2014 and 2019 to the German National Reference Center (NRC) for diagnostic reasons (NRC strain set), and 1143 isolates from a Tertiary Care University Center in Saarland, Germany (non-NRC strain set), were evaluated using antibiotic susceptibility testing and ribotyping. In the NRC strain set, RT027 overtook RT001, the main RT found in the preceding studies, and dominated with 36.2%, followed by RT001 (13.3%), and RT014 (8.5%). Of note, since 2016 a constant decrease of RT027 could be noticed. In the non-NRC strain set a large strain diversity was present with RT014 (18%) and RT001 (8.9%) being most prevalent. In NRC samples, resistance towards metronidazole, vancomycin, moxifloxacin, clarithromycin and rifampicin was 2.7%, 0%, 57.1%, 53.2% and 19.2%, respectively. Metronidazole resistance was almost exclusively found in RT027 isolates. Rifampicin resistance was also observed predominantly in isolates of RT027, constituting an almost four-fold increase, when compared to preceeding studies in this region. In conclusion these data demonstrate that RT027 is a driver for rifampicin and metronidazole resistance, underlining the importance of continuous surveillance efforts.

Simple Questionnaires to Improve Pooling Strategies for SARS-CoV-2 Laboratory Testing.

Background: Liberal PCR testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is key to contain the coronavirus disease 2019 (COVID-19) pandemic. Combined multi-sample testing in pools instead of single tests might enhance laboratory capacity and reduce costs, especially in low- and middle-income countries. Objective: The purpose of our study was to assess the value of a simple questionnaire to guide and further improve pooling strategies for SARS-CoV-2 laboratory testing. Methods: Pharyngeal swabs for SARS-CoV-2 testing were obtained from healthcare and police staff, hospital inpatients, and nursing home residents in the southwestern part of Germany. We designed a simple questionnaire, which included questions pertaining to a suggestive clinical symptomatology, recent travel history, and contact with confirmed cases to stratify an individual's pre-test probability of having contracted COVID-19. The questionnaire was adapted repeatedly in face of the unfolding pandemic in response to the evolving epidemiology and observed clinical symptomatology. Based on the response patterns, samples were either tested individually or in multi-sample pools. We compared the pool positivity rate and the number of total PCR tests required to obtain individual results between this questionnaire-based pooling strategy and randomly assembled pools. Findings: Between March 11 and July 5, 2020, we processed 25,978 samples using random pooling (n = 6,012; 23.1%) or questionnaire-based pooling (n = 19,966; 76.9%). The overall prevalence of SARS-CoV-2 was 0.9% (n = 238). Pool positivity (14.6% vs. 1.2%) and individual SARS-CoV-2 prevalence (3.4% vs. 0.1%) were higher in the random pooling group than in the questionnaire group. The average number of PCR tests needed to obtain the individual result for one participant was 0.27 tests in the random pooling group, as compared to 0.09 in the questionnaire-based pooling group, leading to a laboratory capacity increase of 73% and 91%, respectively, as compared to single PCR testing. Conclusions: Strategies that combine pool testing with a questionnaire-based risk stratification can increase laboratory testing capacities for COVID-19 and might be important tools, particularly in resource-constrained settings.

Epidemiology of Clostridium difficile in Germany based on a single center long-term surveillance and German-wide genotyping of recent isolates provided to the advisory laboratory for diagnostic reasons.

Epidemiology of Clostridium difficile is characterized by worldwide increase of C. difficile infections (CDI) and the emergence of new epidemic outbreak strains with the capacity for global spreading. Long-term local surveillance at the University of Saarland Medical Center between 2000 and 2013 shows that the incidence rate of laboratory-confirmed CDI was influenced by local epidemiology as well as by testing strategies. Since 2008, molecular typing of C. difficile was regularly performed for symptomatic hospitalized patients by surface-layer protein A sequence typing (slpAST), which is an established highly standardized technique for genotyping of C. difficile. The results were assigned to known ribotypes for better comparison to international data. It could be demonstrated that distribution of genotypes was different between age groups. Older patients were predominantly infected with ribotype 001 and 027, whereas ribotype 027 was not detected in the pediatric population. Molecular typing of German isolates sent to the advisory laboratory between 2011 and 2013 revealed that ribotype 027 is present with high percentages in most German regions except for the very North. In conclusion, optimized testing of all hospitalized patients with diarrhea should be generally implemented to avoid under-diagnosis of C. difficile infection. Ribotype 027 is highly prevalent in Germany, but its infections are restricted to older patients, while absent in children. Molecular typing of suspected hospital outbreaks and of patients with severe or recurrent disease may help to better understand virulence and epidemic spreading of C. difficile.

A pseudo-outbreak due to Acinetobacter species (GIM-1) contamination of the pneumatic transport system of a Large University Hospital.

OBJECTIVE: To establish the source and contamination routes resulting in positive clinical and surveillance microbiological cultures with carbapenem-resistant, GIM-1 metallo-ß-lactamase-positive Acinetobacter pitii and Acinetobacter radioresistens from 21 patients in 8 departments. DESIGN: Retrospective, descriptive study. SETTING: A 1,300-bed tertiary care academic medical facility consisting of 90 buildings linked by a pneumatic transport system (PTS). METHODS: Microbiological workup of the cluster strains included matrix-assisted laser desorption/ionization time-of-flight species identification, phenotypic carbapenemase tests, polymerase chain reaction-based genotyping of carbapenemase, and pulsed-field gel electrophoresis. Outbreak management procedures were employed according to institutional regulations. RESULTS: The rarity of GIM-1 Acinetobacter species in the hospital and region, the lack of epidemiological links between patients, and the fact that in some patients the apparent colonization was clearly nonnosocomial prompted the suspicion of a pseudo-outbreak. Numerous environmental cultures were positive for GIM-1-positive Acinetobacter (including archived sample requisition forms, PTS capsules, cultures from line-diverter and dispenser stations, and sterilized transport capsules following PTS delivery). Moreover, it was observed that condensation fluid from subterranean PTS tubing resulted in water entry in PTS capsules, possibly conferring specimen contamination. After extensive system disinfection, environmental surveys of the PTS were negative, and no further positive patient specimens were encountered. CONCLUSIONS: This is the first report of a PTS-associated pseudo-outbreak. The large number of falsely positive patient-related specimens in conjunction with the potential hazard of airborne and contact spread of multidrug-resistant microorganisms (in this case, GIM-1 carbapenem-resistant Acinetobacter species) underscores the need for implementation of infection control-based monitoring and operating procedures in a hospital PTS.

Clinical and radiological findings in long-term intracranial pressure monitoring.

BACKGROUND: Advantages of telemetric devices for long-term intracranial pressure (ICP) measurement have been mentioned several times in the literature. However, descriptions of associated complications are lacking. Therefore, the presented observational study focused on clinical and radiological findings after insertion of an intraparenchymal telemetric ICP monitor. METHODS: Between April 2010 and February 2013, 185 telemetric ICP catheters were implanted for diagnostic purposes. All patients were clinically followed. Radiological, microbiological and clinical data were analysed. RESULTS: One brain abscess (0.5 %) and two cutaneous infections (1.1 %) occurred in 185 patients. Staphylococcus spp. could be detected in all cases. Six patients (3.2 %) suffered from single new-onset seizures and one patient (0.5 %) from a temporary hemiparesis. Intracerebral haemorrhages occurred in 15.6 %, most of the time as small punctate bleedings. Perifocal oedematous reactions surrounding inserted telemetric catheters could be observed in 46.9 %. Multiple imaging studies revealed a tendency of complete oedema resolution over time. CONCLUSIONS: Infectious as well as haemorrhagic complication rates are well comparable with the common literature. The long-term implantation of an ICP probe does not seem to increase the risk of wound infections or brain abscess formation. Surprisingly, very high numbers of oedematous reactions after insertion of the intraparenchymal ICP monitor were seen. Reasons therefore could only be speculated upon.

Methicillin-resistant Staphylococcus aureus in Saarland, Germany: a statewide admission prevalence screening study.

BACKGROUND: The screening of hospital admission patients for methicillin resistant Staphylococcus aureus (MRSA) is of undisputed value in controlling and reducing the overall MRSA burden; yet, a concerted parallel universal screening intervention throughout all hospitals of an entire German Federal State has not yet been performed. METHODOLOGY/PRINCIPAL FINDINGS: During a four-week period, all 24 acute care hospitals of the State of Saarland participated in admission prevalence screening. Overall, 436/20,027 screened patients revealed MRSA carrier status (prevalence, 2.2/100 patients) with geriatrics and intensive care departments associated with highest prevalence (7.6/100 and 6.3/100, respectively). Risk factor analysis among 17,975 admission patients yielded MRSA history (OR, 4.3; CI95 2.7-6.8), a skin condition (OR, 3.2; CI95 2.1-5.0), and/or an indwelling catheter (OR, 2.2; CI95 1.4-3.5) among the leading risks. Hierarchical risk factor ascertainment of the six risk factors associated with highest odd's ratios would require 31% of patients to be laboratory screened to allow for detection of 67% of all MRSA positive admission patients in the State. CONCLUSIONS/SIGNIFICANCE: State-wide admission prevalence screening in conjunction with risk factor ascertainment yields important information on the distribution of the MRSA burden for hospitals, and allows for data-based decisions on local or institutional MRSA screening policies considering risk factor prevalence and expected MRSA identification rates.

Activity of the two-component regulatory system CiaRH in Streptococcus pneumoniae R6.

The two-component regulatory system CiaRH of Streptococcus pneumoniae affects a variety of processes such as competence development, autolysis, bacteriocin production, host colonization, and virulence. While the targets of the regulator CiaR are known, the role of phosphorylation in CiaR regulation has not been defined. To address this issue, the presumed phosphorylation site of CiaR, aspartic acid at position 51, was replaced by alanine. The mutant CiaRD51A protein was no longer able to activate CiaR-dependent promoters, strongly suggesting that the phosphorylated form of CiaR is active in regulation. However, depending on the growth medium, inactivation of the kinase gene ciaH resulted in a subtle increase of CiaR-dependent promoter activities or in a strong reduction. Therefore, CiaH may act as a kinase or phosphatase and CiaR is apparently able to obtain its phosphate independently of CiaH. On the other hand, promoter measurements in cells with an intact CiaRH system demonstrated a high, nearly constitutive, expression level of the CiaR regulon independent from the growth medium. Thus, in contrast to many other two-component regulatory systems, CiaRH has apparently evolved to maintain high levels of gene expression under a variety of conditions rather than responding strongly to a signal.

Identification of the genes directly controlled by the response regulator CiaR in Streptococcus pneumoniae: five out of 15 promoters drive expression of small non-coding RNAs.

The two-component regulatory system CiaRH of Streptococcus pneumoniae has been implicated in beta-lactam resistance, maintenance of cell integrity, competence and virulence, but the genes that are regulated directly by the system have not been defined. Using transcriptional mapping, in vitro CiaR binding, and in vivo analysis of CiaR-mediated regulation, 15 promoters were identified to be directly controlled by the response regulator CiaR. A direct repeat, TTTAAG-N5-TTTAAG, was found to be essential for CiaR binding and regulation. It is present, either completely or with subtle changes, in all promoter regions. Fourteen promoters of the regulon are activated by CiaR, and one was found to be controlled negatively. The genes that are transcribed from these promoters included ciaRH, loci that are predicted to be involved in the modification of teichoic acids (lic), in sugar metabolism (mal, man), stress response (htrA), chromosome segregation (parB), protease maturation (ppmA) and unknown functions. Remarkably, the five strongest promoters of the CiaR regulon drive expression of small RNAs. These small RNAs, designated csRNAs for cia-dependent small RNAs, are non-coding, between 87 and 151 nt in size, and show a high degree of similarity to each other. The analysis of deletion mutants in the csRNA genes revealed that csRNA4 and csRNA5 affect stationary-phase autolysis. The identification of five small non-coding regulatory RNAs opens new perspectives to approach the physiological role of the CiaRH two-component regulatory system.

A new integrative reporter plasmid for Streptococcus pneumoniae.

A new promoter probe system for Streptococcus pneumoniae has been developed that allows stable genomic integration of promoters cloned in front of a promoterless hybrid beta-galactosidase gene consisting of translation initiation signals of the protease gene htrA of S. pneumoniae fused to a truncated Escherichia colibeta-galactosidase gene lacZ. Chromosomal insertions of promoter-lacZ fusions are directed to the endogenous beta-galactosidase gene bgaA, thereby abolishing background beta-galactosidase activity. The new system was tested by measuring beta-galactosidase activity directed by the two promoters of the early competence genes comA and comC. The new integrative plasmid offers several advantages compared with existing systems and is especially suited for stable integration of small promoter fragments to conduct mutagenesis or deletion studies.

A functional dlt operon, encoding proteins required for incorporation of d-alanine in teichoic acids in gram-positive bacteria, confers resistance to cationic antimicrobial peptides in Streptococcus pneumoniae.

Streptococcus pneumoniae is one of the few species within the group of low-G +C gram-positive bacteria reported to contain no d-alanine in teichoic acids, although the dltABCD operon encoding proteins responsible for d-alanylation is present in the genomes of two S. pneumoniae strains, the laboratory strain R6 and the clinical isolate TIGR4. The annotation of dltA in R6 predicts a protein, d-alanine-d-alanyl carrier protein ligase (Dcl), that is shorter at the amino terminus than all other Dcl proteins. Translation of dltA could also start upstream of the annotated TTG start codon at a GTG, resulting in the premature termination of dltA translation at a stop codon. Applying a novel integrative translation probe plasmid with Escherichia coli 'lacZ as a reporter, we could demonstrate that dltA translation starts at the upstream GTG. Consequently, S. pneumoniae R6 is a dltA mutant, whereas S. pneumoniae D39, the parental strain of R6, and Rx, another derivative of D39, contained intact dltA genes. Repair of the stop codon in dltA of R6 and insertional inactivation of dltA in D39 and Rx yielded pairs of dltA-deficient and dltA-proficient strains. Subsequent phenotypic analysis showed that dltA inactivation resulted in enhanced sensitivity to the cationic antimicrobial peptides nisin and gallidermin, a phenotype fully consistent with those of dltA mutants of other gram-positive bacteria. In addition, mild alkaline hydrolysis of heat-inactivated whole cells released d-alanine from dltA-proficient strains, but not from dltA mutants. The results of our study suggest that, as in many other low-G+C gram-positive bacteria, teichoic acids of S. pneumoniae contain d-alanine residues in order to protect this human pathogen against the actions of cationic antimicrobial peptides.