Rapid screening of benzyl dodecyl dimethyl ammonium bromide co-metabolic degrading bacteria based on NIR hyperspectral imaging technology.

As a typical disinfectant, the use of benzyl dodecyl dimethyl ammonium bromide (BDAB) has dramatically increased since the emergence of SARS-CoV-2, posing a threat to environmental balance and human health. Screening BDAB co-metabolic degrading bacteria is required for efficient microbial degradation. Conventional methods for screening co-metabolic degrading bacteria are laborious and time-consuming, especially when the number of strains is large. This study aimed to develop a novel method for the rapid screening of BDAB co-metabolic degrading bacteria from the cultured solid medium using near-infrared hyperspectral imaging (NIR-HSI) technology. Based on NIR spectra, the concentration of BDAB in the solid medium can be well predicted by partial least squares regression (PLSR) models, non-destructively and rapidly, with Rc2 > 0.872 and Rcv2 > 0.870. The results show that the predicted BDAB concentrations decrease after degrading bacteria utilization, comparing with the regions where no degrading bacteria grew. The proposed method was applied to directly identify the BDAB co-metabolic degrading bacteria cultured on the solid medium, and two kinds of co-metabolic degrading bacteria RQR-1 and BDAB-1 were correctly identified. This method provides a high-efficiency method for screening BDAB co-metabolic degrading bacteria from a large number of bacteria.

Derivatization of sialylated glycopeptides plus based sialoglycopeptides enrichment using cation exchange media.

Glycosylation is one of the most important post-translational modifications. However, the characterizations of glycopeptides, especially the negatively charged sialoglycopeptides that are associated with various diseases, remain challenging, due to the co-existence with high abundant peptides and the low ionization efficiency of sialoglycopeptides resulting from the carboxyl groups. Therefore, it is essential to develop an efficient enrichment method for sialoglycopeptides. Here, we present a novel derivatization-based enrichment method that can (i) identify linkage isomers of sialic acids by generating mass difference, (ii) unify the net charge of peptides into zero, and (iii) introduce positive charges to sialoglycopeptides by conjugating quaternary ammonium with sialic acid. The derivatization, termed derivatization of sialylated glycopeptides plus (DOSG+), enables efficient enrichment through electrostatic interaction using weak cation exchange (WCX) media. DOSG+ -based WCX enrichment was validated and optimized with samples derived from bovine fetuin. Peptides were removed efficiently (recovery rate <1%). The signal intensity of a selected model sialoglycopeptide was increased by ∼30% (suggesting recovery rate >100%). The method was employed on human alpha-1 acid glycoprotein (AGP), and recombinant human erythropoietin (EPO), demonstrating the application of DOSG+ -based WCX enrichment on complexed N-linked and O-linked sialoglycopeptides. The method is simple, efficient, and targets small-scale sialoglycopeptide enrichment.

Quaternary ammonium disinfectants and antiseptics: tolerance, resistance and potential impact on antibiotic resistance.

BACKGROUND: Due to the substantial increase in the use of disinfectants containing quaternary ammonion compounds (QACs) in healthcare and community settings during the COVID-19 pandemic, there is increased concern that heavy use might cause bacteria to develop resistance to QACs or contribute to antibiotic resistance. The purpose of this review is to briefly discuss the mechanisms of QAC tolerance and resistance, laboratory-based evidence of tolerance and resistance, their occurrence in healthcare and other real-world settings, and the possible impact of QAC use on antibiotic resistance. METHODS: A literature search was conducted using the PubMed database. The search was limited to English language articles dealing with tolerance or resistance to QACs present in disinfectants or antiseptics, and potential impact on antibiotic resistance. The review covered the period from 2000 to mid-Jan 2023. RESULTS: Mechanisms of QAC tolerance or resistance include innate bacterial cell wall structure, changes in cell membrane structure and function, efflux pumps, biofilm formation, and QAC degradation. In vitro studies have helped elucidate how bacteria can develop tolerance or resistance to QACs and antibiotics. While relatively uncommon, multiple episodes of contaminated in-use disinfectants and antiseptics, which are often due to inappropriate use of products, have caused outbreaks of healthcare-associated infections. Several studies have identified a correlation between benzalkonium chloride (BAC) tolerance and clinically-defined antibiotic resistance. The occurrence of mobile genetic determinants carrying multiple genes that encode for QAC or antibiotic tolerance raises the concern that widespread QAC use might facilitate the emergence of antibiotic resistance. Despite some evidence from laboratory-based studies, there is insufficient evidence in real-world settings to conclude that frequent use of QAC disinfectants and antiseptics has promoted widespread emergence of antibiotic resistance. CONCLUSIONS: Laboratory studies have identified multiple mechanisms by which bacteria can develop tolerance or resistance to QACs and antibiotics. De novo development of tolerance or resistance in real-world settings is uncommon. Increased attention to proper use of disinfectants is needed to prevent contamination of QAC disinfectants. Additional research is needed to answer many questions and concerns related to use of QAC disinfectants and their potential impact on antibiotic resistance.

GS-2: A Novel Broad-Spectrum Agent for Environmental Microbial Control.

The environmental control of microbial pathogens currently relies on compounds that do not exert long-lasting activity on surfaces, are impaired by soil, and contribute to the growing problem of antimicrobial resistance. This study presents the scientific development and characterization of GS-2, a novel, water-soluble ammonium carboxylate salt of capric acid and L-arginine that demonstrates activity against a range of bacteria (particularly Gram-negative bacteria), fungi, and viruses. In real-world surface testing, GS-2 was more effective than a benzalkonium chloride disinfectant at reducing the bacterial load on common touch-point surfaces in a high-traffic building (average 1.6 vs. 32.6 CFUs recovered from surfaces 90 min after application, respectively). Toxicology testing in rats confirmed GS-2 ingredients were rapidly cleared and posed no toxicities to humans or animals. To enhance the time-kill against Gram-positive bacteria, GS-2 was compounded at a specific ratio with a naturally occurring monoterpenoid, thymol, to produce a water-based antimicrobial solution. This GS-2 with thymol formulation could generate a bactericidal effect after five minutes of exposure and a viricidal effect after 10 min of exposure. Further testing of the GS-2 and thymol combination on glass slides demonstrated that the compound retained bactericidal activity for up to 60 days. Based on these results, GS-2 and GS-2 with thymol represent a novel antimicrobial solution that may have significant utility in the long-term reduction of environmental microbial pathogens in a variety of settings.

[Migration, Transformation, and Toxicity of Quaternary Ammonium Antimicrobial Agents in the Environment].

Quaternary ammonium compounds (QACs) are one type of widely used cationic biocide, and their usage amount is growing rapidly due to the flu and COVID-19 pandemic. Many QACs were released into the environment in or after the course of their use, and thus they were widely detected in water, sediment, soil, and other environmental media. QACs have stronger surface activity and non-specific biotoxicity, which poses a potential threat to the ecosystem. In this study, the environmental fate and potential toxicity of QACs were documented in terms of their migration and transformation process, biological toxicity effects, and the main mechanisms of bacterial resistance to QACs. Aerobic biodegradation was the main natural way of eliminating QACs in the environment, and the reaction was mainly initiated by the hydroxylation of C atoms at different positions of QACs and finally mineralized to CO2and H2O through decarboxylation, demethylation, and ß-oxidation reaction. Toxicological studies showed that QACs at environmental concentrations could not pose acute toxicity to the selected biotas but threatened the growth and reproduction of aquatic organisms like Daphnia magna. Their toxicity effects depended on their molecular structure, the tested species, and the exposed durations. Additionally, our team first investigated the toxicity effects and mechanisms of QACs toward Microcystis aeruginosa, which showed that QACs depressed the algae growth through the denaturation of photosynthetic organelles, suppression of electron transport, and then induction of cell membrane damage. In the environment, the concentrations of QACs were always lower than their bactericidal concentrations, and their degradation could induce the formation of a concentration gradient, which facilitated microbes resistant to QACs. The known resistance mechanisms of bacteria to QACs mainly included the change in cell membrane structure and composition, formation of biofilm, overexpression of the efflux pump gene, and acquisition of resistance genes. Due to the similar targets and mechanisms, QACs could also induce the occurrence of antibiotic resistance, mainly through co-resistance and cross-resistance. Based on the existing data, future research should emphasize the toxicity effect and the potential QACs resistance mechanism of microorganisms in real environmental conditions.

Smart Approach for the Design of Highly Selective Aptamer-Based Biosensors.

Aptamers are chemically synthesized single-stranded DNA or RNA oligonucleotides widely used nowadays in sensors and nanoscale devices as highly sensitive biorecognition elements. With proper design, aptamers are able to bind to a specific target molecule with high selectivity. To date, the systematic evolution of ligands by exponential enrichment (SELEX) process is employed to isolate aptamers. Nevertheless, this method requires complex and time-consuming procedures. In silico methods comprising machine learning models have been recently proposed to reduce the time and cost of aptamer design. In this work, we present a new in silico approach allowing the generation of highly sensitive and selective RNA aptamers towards a specific target, here represented by ammonium dissolved in water. By using machine learning and bioinformatics tools, a rational design of aptamers is demonstrated. This "smart" SELEX method is experimentally proved by choosing the best five aptamer candidates obtained from the design process and applying them as functional elements in an electrochemical sensor to detect, as the target molecule, ammonium at different concentrations. We observed that the use of five different aptamers leads to a significant difference in the sensor's response. This can be explained by considering the aptamers' conformational change due to their interaction with the target molecule. We studied these conformational changes using a molecular dynamics simulation and suggested a possible explanation of the experimental observations. Finally, electrochemical measurements exposing the same sensors to different molecules were used to confirm the high selectivity of the designed aptamers. The proposed in silico SELEX approach can potentially reduce the cost and the time needed to identify the aptamers and potentially be applied to any target molecule.

Effect of plastic pollution in soil properties and growth of grass species in semi-arid regions: a laboratory experiment.

Since the year 2020, the use of plastic as a strategy to mitigate the spread of COVID-19 disease has been given substantial attention. Global environmental contamination of plastic creates waste and is a known threat to soil ecosystems as a main sink of microplastics. However, there is still considerable uncertainty about microplastics controlling soil properties alteration. Therefore, we carried out an incubation experiment with soil and Carex stenophylla Wahlenb., which are the dominant soil and grass species in semi-arid regions. We investigated the effect of polymer polyethylene terephthalate (PET) concentrations (0%, 1%, 3%, and 5%) on C. stenophylla growth and soil ammonium-N and nitrate-N, organic matter content, pH, soil aggregates, and soil respiration. When soils were exposed to PET microplastics, fewer seeds germinated (62.8 ± 32%) but not significantly (p value > 0.05) when soils were treated to 0, 1, 3, and 0.5% PET. Shoot height was also not effectively reduced with PET. The soil pH was considerably lower when exposed to higher PET compared to all other treatments with the soil exposed to 5% w/w PET for both unplanted and planted, being 0.84 and 0.54 units, respectively, lower than the controls. The soil microbial respiration under exposure to PET was considerably increased in comparison to control samples. Moreover, the presence of PET resulted in potential alterations of soil stability, and with PET present soil stability increased. In conclusion, PET microplastics cannot significantly affect the development of C. stenophylla but could affect crucial soil properties. In addition, changes occurred with increased variability in soil ammonium-N and nitrate-N, particularly at a high PET ratio.

The first detection of quaternary ammonium compounds in breast milk: Implications for early-life exposure.

BACKGROUND: Quaternary ammonium compounds (QACs), commonly used in cleaning, disinfecting, and personal care products, have recently gained worldwide attention due to the massive use of disinfectants during the COVID-19 pandemic. However, despite extensive use of these chemicals, no studies have focused on the analysis of QACs in human milk, a major route of exposure for infants. OBJECTIVE: Our objectives were to identify and measure QACs in breast milk and evaluate early-life exposure to this group of compounds for nursing infants. METHODS: Eighteen QACs, including 6 benzylalkyldimethyl ammonium compounds (BACs, with alkyl chain lengths of C8-C18), 6 dialkyldimethyl ammonium compounds (DDACs, C8-C18), and 6 alkyltrimethyl ammonium compounds (ATMACs, C8-C18), were measured in breast milk samples collected from U.S. mothers. Daily lactational intake was estimated based on the determined concentrations for 0-12 month old nursing infants. RESULTS: Thirteen of the 18 QACs were detected in breast milk and 7 of them were found in more than half of the samples. The total QAC concentrations (ΣQAC) ranged from 0.33 to 7.4 ng/mL (median 1.5 ng/mL). The most abundant QAC was C14-BAC with a median concentration of 0.45 ng/mL. The highest median ΣQAC estimated daily intake (EDI) was determined for <1-month old infants based on the average (using the median concentration) and high (using the 95th percentile concentration) exposure scenarios (230 and 750 ng/kg body weight/day, respectively). SIGNIFICANCE: Our findings provide the first evidence of the detection of several QACs in breast milk and identify breastfeeding as an exposure pathway to QACs for nursing infants. IMPACT STATEMENT: Our findings provide the first evidence of QAC occurrence in breast milk and identify breastfeeding as one of the exposure pathways to QACs for nursing infants.

Evaluation of virucidal activity of residual quaternary ammonium-treated surfaces on SARS-CoV-2.

BACKGROUND: The COVID-19 pandemic has had an unprecedented impact on global health and the world's economies. Proliferation of virulent and deadly SARS-CoV-2 variants require effective transmission mitigation strategies. Under reasonable environmental conditions, culturable and infectious SARS-CoV-2 can survive on contaminated fomites from hours to months. In the present study we evaluated a surface-anchored polymeric quaternary ammonium antimicrobial to help reduce fomite transmission of SARS-CoV-2 from contaminated surfaces. METHODS: Two studies were performed on antimicrobial pre-treated metal disks in March 2020 by two independent Biosafety Level III (BSL-3) equipped laboratories in April 2020. These facilities were in Belgium (the Rega Medical Research Institute) and Australia (the Peter Doherty Institute) and independently applied quantitative carrier-based methodologies using the authentic SARS-CoV-2 isolates (hCoV-19/Australia/VIC01/2020, hCoV-19/Belgium/GHB-03021/2020). RESULTS: Residual dry tests were independently conducted at both facilities and demonstrated sustained virion destruction (108.23 TCID50/carrier GHB-03021 isolate, and 103.66 TCID50/carrier VIC01 isolate) 1 hour (drying) + 10 minutes after inoculation. Reductions are further supported by degradation of RNA on antimicrobial-treated surfaces using qRT-PCR. CONCLUSIONS: Using a polymeric quaternary ammonium antimicrobial (EPA/PMRA registered) the results independently support a sustained antiviral effect via SARS-CoV-2 virion destruction and viral RNA degradation. This indicates that silane-anchored quaternary ammonium compound (SiQAC-18) treated surfaces could play an important role in mitigating the communicability and fomite transmission of SARS-CoV-2.

Anti-SARS-CoV-2 Activity of Surgical Masks Infused with Quaternary Ammonium Salts.

The SARS-CoV-2 pandemic has highlighted the need for protective and effective personal protective equipment (PPE). Research has shown that SARS-CoV-2 can survive on personal protective equipment, such as commonly used surgical masks. Methods are needed to inactivate virus on contaminated material. We show here that embedding viral-disinfecting compounds during the manufacturing of surgical masks inactivates a high dose (up to 1 × 105 pfu) of live, authentic SARS-CoV-2 within minutes.

Safe evacuation system for pneumoperitoneum and surgical smoke in laparoscopic surgery using a water trap in times of COVID-19 / Sistema de evacuación seguro de neumoperitoneo y humo quirúrgico en cirugía laparoscópica mediante trampa de agua en tiempos de COVID-19

INTRODUCCIÓN: La reciente pandemia por SARS-CoV-2 (COVID-19) ha hecho resurgir preocupación sobre la exposición inadvertida del equipo quirúrgico a agentes infecciosos transmisibles por vía aérea durante el acto quirúrgico. El objetivo de este trabajo es describir la confección de un sistema de filtrado simple y de bajo costo que permite reducir el riesgo de exposición al virus, particularmente en el proceso de aspiración, recambio y remoción del neumoperitoneo en cirugía laparoscópica. MATERIALES Y MÉTODO: Se diseñó e implementó un circuito cerrado de evacuación y de filtrado del neumoperitoneo en cirugías ginecológicas laparoscópicas en un centro de salud terciario. El circuito incluye un filtro HEPA (High Efficiency Particulate Air) y una trampa de vacío que contiene una solución de inactivación en base a amonio cuaternario o hipoclorito de sodio. RESULTADOS: Desde su implementación se han realizado 17 cirugías laparoscópicas ginecológicas por diversas patologías. Dos de ellas fueron en pacientes Covid-19 (+). A la fecha no se han reportado contagios en el equipo médico que participó en la cirugía. CONCLUSIONES: Es posible implementar un sistema de evacuación del neumoperitoneo en cirugía laparoscópica presumiblemente eficaz en minimizar el riesgo de exposición al virus SARS-COV-2 (Covid-19). Su bajo costo lo hace especialmente recomendable en países en vías de desarrollo.

INTRODUCTION: The recent SARS-CoV-2 (COVID-19) pandemics has raised concern on the incidental exposition of health team to air transmissible infectious agents during surgeries. The main goal of this work is to communicate a simple and low-cost filtering system allowing to reduce the risk of contagion related to the virus, associated with pneumoperitoneum removal during surgical laparoscopy. METHODS: A closed circuit of gas removal and filtering was developed and implemented in laparoscopic gynecologic procedures at a tertiary teaching hospital. The circuit included an HEPA (High Efficiency Particulate Air) filter and a vacuum trap containing an inactivating solution based on quaternary ammonium or sodium hypochlorite. RESULTS: Since its introduction, seventeen laparoscopic surgeries have been carried out for different gynecologic pathologies. Two of them in Covid (+) cases. To date, no contagion has been reported among health teammates participating in these surgeries. CONCLUSIONS: It is possible to implement a pneumoperitoneum evacuation system in laparoscopic surgery presumably effective in minimizing the risk of exposure to the SARS-COV-2 virus (Covid-19). Its low cost makes it especially recommended in developing countries.