Metagenomic insights into microbial contamination in critical healthcare environments and the efficacy of a novel "HLE" disinfectant.

BACKGROUND: Bacterial contamination on inanimate clinical surfaces is directly linked to severe health problems, especially those caused by multidrug resistant (MDR) pathogens. Here, we evaluated the microbial burden in these environments and tested the efficacy of a novel HLE disinfectant solution. METHODS: Microbial contamination of healthcare surfaces [Intensive Care Unit (ICU), Long Period Hospitalization Room (LPHR) and Otolaryngology Consultation (OC)] and the efficacy of HLE disinfectant solution were determined analyzing the viable counts on general and selective media, and also by molecular studies focused on metagenomic and specific qPCR. RESULTS: Different contamination loads were detected with LPHR showing the highest contamination. Treatment with the HLE disinfectant solution curbed the spread of well-adapted pathogens on touched surfaces (ICU, LPHR and OC). Metagenomic analysis of microbial diversity of the Patient Table (most contaminated surface in LPHR) revealed the presence of mainly A. johnsonii and P. putida. Furthermore, functional annotation of toxin, virulence and antibiotic resistance sequences showed a high diversity of Acinetobacter spp. and Pseudomonas spp. In this context, specific qPCR analysis confirmed the efficacy of HLE disinfectant solution against the most prevalent and critical pathogens Pseudomonas sp. and Acinetobacter sp. achieving their complete eradication. CONCLUSION: Given the persistence of detrimental resistant pathogens, the application of HLE disinfection solution could be a highly beneficial and effective option -used either alone or in combination-for infection prevention and control with the aim to eliminate microbial pathogens and their genes from contaminated contact-surfaces and thus limit the spread to humans and other ecological niches.

Novel combination of nanoparticles and metallo-ß-lactamase inhibitor/antimicrobial-based formulation to combat antibiotic resistant Enterococcus sp. and Pseudomonas sp. strains.

Nanotechnology presents an innovative strategy to combat the spread of antibiotic resistant bacteria and their resistance genes throughout different ecosystems. To address this challenge, nanoparticles (silver, gold, zinc and copper) alone or in combination with metallo-ß-lactamase inhibitor/antimicrobial-based formulation (EDTA/HLE) showed antimicrobial activity against antibiotic resistant Enterococcus sp. and Pseudomonas sp. strains. Furthermore, the observed synergistic effect was detected notably for silver, zinc or copper nanoparticles with EDTA (ethylenediaminetetraacetic acid) and silver nanoparticles with HLE against planktonic Enterococcus sp. strains, or gold nanoparticles+EDTA or HLE against Pseudomonas sp. Regarding activity against bacterial biofilms, zinc nanoparticles combined with either of the reagents caused strong inhibition of developing biofilms of antibiotic resistant Enterococcus sp. Pseudomonas sp. strains, while preformed biofilms were mainly inhibited by silver nanoparticles+reagent. Microscopic analyses confirmed that the antimicrobial activity of nanoparticles was caused by adsorption to the bacterial cell surface, and further enhanced by chelating agents. Hence, we can conclude that nanoparticles+EDTA or HLE could represent a good alternative to limit the spread of antibiotic resistant bacteria in the food chain and the environment.

Antibacterial and antibiofilm effects of essential oil components, EDTA and HLE disinfectant solution on Enterococcus, Pseudomonas and Staphylococcus sp. multiresistant strains isolated along the meat production chain.

The spread of multidrug resistant (MDR) bacteria and resistance genes along the food chain and the environment has become a global, but silent pandemic. To face this challenge, it is of outmost importance to develop efficient strategies to reduce potential contamination by these agents. In the present study, 30 strains of Enterococcus sp., Staphylococcus sp. and Pseudomonas sp. isolated from various surfaces throughout the meat production chain in a goat and lamb slaughterhouse were characterized as MDR bacteria harboring several antibiotic resistance genes (ARGs). The antimicrobial efficacy of natural essential oil components "EOCs" (carvacrol "CA," cinnamaldehyde "CIN," eugenol "EU," geraniol "GE," limonene "LI" and thymol "TH"), HLE disinfectant solution (3-6% H2O2; 2.2-4.4% lactic acid and 12.5-25 mM EDTA in water) and EDTA was tested against these MDR bacteria. Results showed that Minimum Inhibitory Concentrations (MIC) were compound and strain dependent. In addition, the synergistic effect of these antimicrobials was evaluated at 1/2 MIC. Here our study showed particularly promising results regarding the inhibitory effect at sub-inhibitory concentrations, which were confirmed by the analysis of bacterial growth dynamics over 72 h. Furthermore, the inhibitory effect of EOCs, HLE disinfectant solution and EDTA or their combinations was studied in developing and established biofilms of MDR bacteria obtaining variable results depending on the morphological structure of the tested strain and the phenolic character of the EOCs. Importantly, the combination of EOCs with HLE or EDTA showed particularly positive results given the effective inhibition of biofilm formation. Moreover, the synergistic combinations of EU and HLE/EDTA, TH, CA, GE, LI or CIN + EDTA/HLE caused log reductions in established biofilms of several strains (1-6 log10 CFU) depending on the species and the combination used, with Pseudomonas sp. strains being the most susceptible. Given these results, we propose novel antimicrobial formulations based on the combination of sub-inhibitory concentrations of EOCs and HLE or EDTA as a highly promising alternative to currently used approaches. This novel strategy notably shows great potential to efficiently decrease the emergence and spread of MDR bacteria and ARGs in the food chain and the environment, thus supporting the decrease of resistomes and pathogenesis in clinical and industrial areas while preserving the antibiotic therapeutic action.

Physical and Biological Performance Evaluation of Disinfection Systems for Transportation Vehicles against AI Virus.

To prevent the outbreak of infectious diseases that inflict huge economic and social losses, domestic livestock farms and related facilities have introduced automatic and semiautomatic disinfectant solution-spraying systems for vehicles. However, the facility standards and specifications vary by manufacturer, and no scientific performance evaluation has been conducted. The puropose of this study is to develop physical and biological evaluation methods. Physical and biological appraisals were conducted using two types of disinfection facilities (tunnel- and U-type) and two types of vehicles (passenger car, truck). Water-sensitive paper was used to evaluate the physical performance values for the disinfection facilities. In addition, to assess their biological performance, carriers containing low-pathogenic avian influenza virus were attached to vehicles, and the viral reduction was measured after the vehicles moved through the facility. The tunnel-type had rates of coverage in the range of 70-90% for the passenger car and 60-90% for the truck. At least 4-log virus reduction after spraying for 1-5 min was shown for both vehicles. For the U-type facility evaluation, the coverage rates were in the range of 60-90% for the passenger car and at least 90% for the truck. More than 4-log viral reduction was estimated within a spraying time of 5 min. To reduce viruses on the surface of vehicles by at least 4 log within a short period, the disinfectant solution should cover at least 71% of the pathogens. In conclusion, we were able to assess the physical and biological performance criteria for disinfection facilities aboard transportation vehicles.

Disinfection of gloved hands during the COVID-19 pandemic.

BACKGROUND: Disinfection of gloves can be used during a pandemic situation when performing various procedures on the same patient or when removing personal protective equipment. If performing glove disinfection, there is a need to check the compatibility of gloves with the disinfectant product used. AIM: To test the resistance of nitrile gloves to various disinfectant solutions. METHODS: One hundred percent powder-free nitrile gloves, composed of nitrile butadiene rubber compounds, were exposed to various disinfectants to analyse resistance. The seven most commonly used disinfectant solutions in the healthcare field were selected for testing. The effects of each disinfectant were analysed in comparison with the control group (untreated glove). For tensile testing, the thickness of each test specimen was measured with a micrometer. FINDINGS: Bleach solution decreased the breaking load of gloves, although to a lesser extent than disinfectants that contained ethanol. CONCLUSION: Disinfectants that contain alcohol decrease the breaking load of nitrile gloves.

Analysis of P. aeruginosa disinfectant sensitivity and microbial adhesions to worn cosmetic contact lenses.

PURPOSE: To compare the sensitivity of two genotypes of P. aeruginosa to various disinfectant solutions and analyze the attached bacteria on worn cosmetic contact lenses (cosCLs). METHODS: In this prospective study, healthy volunteers wore etafilcon (brown), nelfilcon (gray), or hilafilcon (black) cosCLs and microbial adhesion analysis was performed. A rub-off test determined pigment dislodgement. Disinfectant sensitivity to Optifree Replenish (Alcon), Optifree Pure Moist (Alcon), Renu Fresh (Bausch & Lomb), and AoSept Plus (Ciba Vision) was tested at various disinfection times and compared between various genotypes and Type III secretion (T3S) system mutants. RESULTS: Of the 1152 cosCLs collected, 364 were culture positive (32%). The highest rate of culture-positive lens was hilafilcon (chi square, P = 0.0001). Hilafilcon also had a significantly greater number of isolates than etafilcon (P < 0.0001). Hilafilcon was the only lens to fail the rub-off test. Cytotoxic strains were significant more resistant to Renu Fresh than were invasive strains, even at 100% of recommended disinfection time (P = 0.0005). Of the tested disinfectants, Renu Fresh was significantly less effective in killing both genotypes of P. aeruginosa compared to AoSept Plus at all time points (25%, 50%, 75%, and 100% recommended disinfection time, P = 0.0001, 0.0001, 0.0005, and 0.0005, respectively). When the T3S system was dysfunctional, mutant strains were all susceptible to disinfectants (P = 0.0001 for both invasive and cytotoxic strains). CONCLUSION: Pseudomonas species is commonly found on cosCLs of asymptomatic individuals. Wearers of cosCLs that dislodge pigments may be predisposed to microbial contamination. Cytotoxic strains are more resistant to disinfectant solutions, especially to Renu Fresh. P. aeruginosa disinfectant resistance requires a functional T3S system.

Influence of chloramine-T disinfection on elastomeric impression stability.

OBJECTIVE: The aim of this study was to evaluate the stability of elastomer through detail reproduction and its dimensional stability (DS) after disinfection with 0.2% chloramine-T. MATERIALS AND METHODS: The elastomeric impression dental materials used in this study were polydimethylsiloxane (Oranwash L), polyvinyl siloxane (Express), polysulfide (Permlastic), and polyether (Impregum Soft). The entire press procedure was performed on a matrix in accordance with the ISO 4823. Detail reproduction was analyzed using an optical microscope (Stereozoom Microscope) over the 20-µm line with 25 mm of length at a magnification of ×4. DS was measured using an optical microscope (Scanning Tunneling Microscope) by subtracting the distance between the lines X and X' over the 20-µm line on the matrix (DM) from the distance between the lines on the impression material (DI) divided by DM and multiplied by 100 establishing the equation: DS = ([DI - DM]/DM) ×100; then, 100% was added to the results of the equation. The detail reproduction values were subsequently subjected to descriptive analysis by percentage (%), and the DS values (%) were submitted to Kolmogorov-Smirnov test, two-way ANOVA (material × disinfectant), and Tukey's test (α = 0.05). RESULTS: All elastomeric impression materials showed 100% of detail reproduction, regardless of the disinfection procedure. Polysulfide (not disinfected) and polysulfide and polydimethylsiloxane (after disinfection with 0.2% chloramine-T) showed the smaller mean values of DS. CONCLUSION: According to the stability properties analyzed, chloramine-T can be used for disinfection of elastomer molds.

Fatal Ingestion of Chlumsky Disinfectant Solution.

A 32-year-old pregnant woman in the 25th week of pregnancy underwent oral glucose tolerance screening at the diabetologist's. Later that day, she was found dead in her apartment possibly poisoned with Chlumsky disinfectant solution (solutio phenoli camphorata). An autopsy revealed chemical burns in the digestive system. The lungs and the brain showed signs of severe edema. The blood of the woman and fetus was analyzed using gas chromatography with mass spectrometry and revealed phenol, its metabolites (phenyl glucuronide and phenyl sulfate) and camphor. No ethanol was found in the blood samples. Both phenol and camphor are contained in Chlumsky disinfectant solution, which is used for disinfecting surgical equipment in healthcare facilities. Further investigation revealed that the deceased woman had been accidentally administered a disinfectant instead of a glucose solution by the nurse, which resulted in acute intoxication followed by the death of the pregnant woman and the fetus.

Comparative Evaluation of Tensile Strength in Die Stone Incorporated with Sodium and Calcium Hypochlorite as Disinfectants: An in vitro Study.

AIM: The aim of this study was to evaluate the tensile strength of die stone incorporated with sodium and calcium hypochlorite as disinfectants. MATERIALS AND METHODS: Two commercially available type IV die stone (Kalrock: Kalabhai Karson Pvt., Ltd and Pearlstone: Asian Chemicals) and two commercially available disinfectant solutions (sodium hypochlorite and calcium hypochlorite: Beachem Laboratory Chemical Private Limited, Chennai and Leo Chem Private Limited, Bengaluru) were used in this study, and the tensile strength was measured using Lloyd's Universal Testing Machine. RESULTS: The results show that incorporating the disinfecting solutions decreases the tensile strength of both products. The effect of decreasing tensile strength on type IV gypsum product is seen more in calcium hypochlorite when compared with sodium hypochlorite disinfecting solution, and the tensile strength of Kalrock specimens is higher than Pearlstone specimens after disinfecting with sodium hypochlorite and calcium hypochlorite solution. The statistical results also show significant results in all the groups when compared with the control group. CONCLUSION: The incorporation of sodium and calcium hypochlorite disinfecting solutions is not an encouraging method for both die materials as it reduces the tensile strength of type IV gypsum product. Tensile strength of Kalstone® die material is superior than Pearlstone® die material after mixing with sodium hypochlorite and calcium hypochlorite. CLINICAL SIGNIFICANCE: According to the recommendations of Americans with Disability Act (ADA) and the Centers for Disease Control and Prevention, disinfecting the whole cast without or minimal changes in physical and mechanical properties was the motto of the study. The tensile strength in type IV gypsum product plays a most important role in retrieval of cast from impression, especially in narrow tooth preparation. This study reveals that incorporating method of disinfecting solutions is not recommended as it reduces the tensile strength.

Bacterial contaminants from frozen puff pastry production process and their growth inhibition by antimicrobial substances from lactic acid bacteria.

Seventy-five bacterial contaminants which still persisted to cleaning system from three puff pastry production lines (dough forming, layer and filling forming, and shock freezing) were identified using 16S rDNA as seven genera of Bacillus, Corynebacterium, Dermacoccus, Enterobacter, Klebsiella, Pseudomonas, and Staphylococcus with detection frequencies of 24.00, 2.66, 1.33, 37.33, 1.33, 2.66, and 30.66, respectively. Seventeen species were discovered while only 11 species Bacillus cereus, B. subtilis, B. pumilus, Corynebacterium striatum, Dermacoccus barathri, Enterobacter asburiae, Staphylococcus kloosii, S. haemolyticus, S. hominis, S. warneri, and S. aureus were detected at the end of production. Based on their abundance, the highest abundance of E. asburiae could be used as a biomarker for product quality. While a low abundance of the mesophile pathogen C. striatum, which causes respiratory and nervous infection and appeared only at the shock freezing step was firstly reported for its detection in bakery product. Six antimicrobial substances (AMSs) from lactic acid bacteria, FF1-4, FF1-7, PFUR-242, PFUR-255, PP-174, and nisin A were tested for their inhibition activities against the contaminants. The three most effective were FF1-7, PP-174, and nisin A exhibiting wide inhibition spectra of 88.00%, 85.33%, and 86.66%, respectively. The potential of a disinfectant solution containing 800 AU/ml of PP-174 and nisin A against the most resistant strains of Enterobacter, Staphylococcus, Bacillus and Klebsiella was determined on artificially contaminated conveyor belt coupons at 0, 4, 8, 12, and 16 hr. The survival levels of the test strains were below 1 log CFU/coupon at 0 hr. The results suggested that a combined solution of PP-174 and nisin A may be beneficial as a sanitizer to inhibit bacterial contaminants in the frozen puff pastry industry.

Influence of two disinfectants methods on disinfection efficiency and model precision of dental impression / 中国实用护理杂志

Objective To iavestigate the influence of two disinfectants methods on disintection efficiency and model precision of dental impression.Methods Alginate impressions,alginate and agar complex impressions and silicon rubber impressions were made according to a unified standard.Chlorinous disinfectant solution and acidic oxidative potential water were used to disinfect the infected impressions.The influence of two disinfectants on disinfection efficiency and model precision of dental impression was compared.Results Two disinfectants for 30 minutes had significant disinfection effect on these three kinds of impressions.The effective rate reached 99.48％.Chlorinous disinfectant solution bad no significant influence on the model precision of alginate impressions and silicon rubber impressions,but had significant influence on the model precision of alginate and agar complex impressions.Acidic oxidative potential water did not influence the model precision of the three dental impressions.Conclusions Acidic oxidative potential water can be used to disinfect the three dental impressions.Chlorinous disinfectant solution can be used to disinfect alginate impressions and silicon rubber impressions,but cannot be used to disinfect agar complex impressions.

Surface detail reproduction and dimensional accuracy of stone models: influence of disinfectant solutions and alginate impression materials

This study compared the surface detail reproduction and dimensional accuracy of stone models obtained from molds disinfected with 2% sodium hypochlorite, 2% chlorhexidine digluconate or 0.2% peracetic acid to models produced using molds which were not disinfected, with 3 alginate materials (Cavex ColorChange, Hydrogum 5 and Jeltrate Plus). The molds were prepared over matrix containing 20-, 50-, and 75-µm lines, performed under pressure with perforated metal tray. The molds were removed following gelation and either disinfected (using one of the solutions by spraying followed by storage in closed jars for 15 min) or not disinfected. The samples were divided into 12 groups (n=5). Molds were filled with dental gypsum Durone IV and 1 h after the start of the stone mixing the models were separated from the tray. Surface detail reproduction and dimensional accuracy were evaluated using optical microscopy on the 50-µm line with 25 mm in length, in accordance with the ISO 1563 standard. The dimensional accuracy results (%) were subjected to ANOVA. The 50 µm-line was completely reproduced by all alginate impression materials regardless of the disinfection procedure. There was no statistically significant difference in the mean values of dimensional accuracy in combinations between disinfectant procedure and alginate impression material (p=0.2130) or for independent factors. The disinfectant solutions and alginate materials used in this study are no factors of choice regarding the surface detail reproduction and dimensional accuracy of stone models.

Este estudo comparou a reprodução de detalhes da superfície e alteração dimensional de modelos de gesso obtidos a partir de moldes desinfetados com hipoclorito de sódio 2%, digluconato de clorexidina 2%, ou ácido peracético 0,2% a modelos confeccionados utilizando moldes que não foram desinfetados com três alginatos (Cavex ColorChange, Hydrogum 5, Jeltrate Plus). Os moldes foram preparados sobre matriz contendo linhas de 20, 50 e 75 µm realizado sob pressão com moldeira de metal perfurada. Os moldes foram removidos após a geleificação e desinfetados (utilizando uma das soluções por pulverização, armazenados em frascos fechados durante 15 min) ou não desinfetados. Assim, as amostras foram divididas em 12 grupos (n=5). Os moldes foram preenchidos com gesso dental Durone IV e uma hora após a manipulação do gesso os modelos foram separados da moldeira. A reprodução de detalhes da superfície e a precisão dimensional foram avaliadas usando microscopia óptica na linha 50 µm com 25 mm de comprimento, de acordo com a norma ISO 1563. Os resultados de precisão dimensional (%) foram submetidos à ANOVA. A linha de 50 µm foi completamente reproduzida por todos os alginatos, independentemente do processo de desinfecção. Não houve diferença estatisticamente significativa nos valores médios de precisão dimensional nas combinações entre procedimento de desinfecção e alginato (p=0,2130), ou para fatores independentes. Soluções desinfetantes e alginatos utilizados neste estudo não são fatores de escolha em relação à reprodução de detalhes da superfície e alteração dimensional de modelos de gesso.