Cationic Biocides Research Articles

Highly Effective Biocides against Pseudomonas aeruginosa Reveal New Mechanistic Insights Across Gram-Negative Bacteria.

Pseudomonas aeruginosa is a major nosocomial pathogen that persists in healthcare settings despite rigorous disinfection protocols due to intrinsic mechanisms conferring resistance. We sought to systematically assess cationic biocide efficacy against this pathogen using a panel of multidrug-resistant P. aeruginosa clinical isolates. Our studies revealed widespread resistance to commercial cationic disinfectants that are the current standard of care, raising concerns about their efficacy. To address this shortcoming, we highlight a new class of quaternary phosphonium compounds that are highly effective against all members of the panel. To understand the difference in efficacy, mechanism of action studies were carried out, which identified a discrete inner-membrane selective target. Resistance selection studies implicated the SmvRA efflux system (a transcriptionally regulated, inner membrane-associated efflux system) as a major determinant of resistance. This system is also implicated in resistance to two commercial bolaamphiphile antiseptics, octenidine and chlorhexidine, which was further validated herein. In sum, this work highlights, for the first time, a discrete inner-membrane specific mechanism for the bolaamphiphile class of disinfectants that contrasts with the prevailing model of indiscriminate membrane interactions of commercial amphiphiles paving the way for future innovations in disinfectant research.

Investigation of the detergency properties of mixtures of biocides and nonionic surfactants using a new simplified hard surface cleaning method

AbstractThe present study explores the cleaning efficacy of a set of nonionic surfactants (linear ethoxylated alcohol, secondary ethoxylated alcohols with 5, 7, and 9 ethoxy groups, glycoside surfactants, polyglycerol surfactants, and an ethoxylated sorbitan monolaurate) combined with cationic biocides—alkyl quaternary ammonium salts. A simple hard surface cleaning methodology was applied, which was shown to discriminate well between poor and good cleaning formulations. In addition to cleaning efficacy, surface aesthetics such as gloss and haze were evaluated together to assess surface streaking caused by a residual surfactant layer. The haziness determination turned out to be the key feature revealing the complex cleaning performance of multi‐component products.

Downstream Effects of the Pandemic? Spatiotemporal Trends of Quaternary Ammonium Compounds in Suspended Particulate Matter of German Rivers

During the SARS-CoV-2 pandemic, the preventive use of antimicrobials such as quaternary ammonium compounds (QACs) surged worldwide. As cationic and surface-active biocides, QACs are only partly removed during wastewater treatment and may cause adverse ecological effects in the downstream environment. To understand the environmental consequences of increased disinfectant use during the pandemic, we investigated spatiotemporal QAC concentration trends in the suspended particulate matter (SPM) of three diverse German rivers. Covering pooled annual SPM samples from 2006-2021 and monthly samples from 2018-2021 collected by the German Environmental Specimen Bank, 31 QACs were quantified by high performance liquid chromatography–mass spectrometry.∑QAC concentrations in annual samples differed by more than tenfold between rivers in the order Saar (average 6.7µg/g) > Rhine (0.9µg/g) > Mulde (0.3µg/g). The strongest potential pandemic imprint was however observed in the Mulde (+67%) and Rhine (+22%). Besides pandemic dynamics, also seasonal variation and mineral content of SPM tentatively affected QAC concentrations. Exceedance of predicted no-effect concentrations for sediment suggest ecotoxicological risks for long-chained QACs already before the pandemic. Our study thus highlights the importance of monitoring the environmental effects of antimicrobial use during pandemics and calls for an urgent minimization of non-essential applications. Environmental implicationThe SARS-CoV-2 pandemic caused a global surge in disinfectant use, including quaternary ammonium compounds (QACs). This raises concerns about their environmental residues, aquatic toxicity and contribution to the (co)selection of antimicrobial resistance in pathogens. To assess these ecotoxicological risks, we investigated spatiotemporal QAC concentration trends in suspended particulate matter of German rivers. While seasonal and spatial fluctuations exceed a potential pandemic “imprint” by a factor of 10, concentrations of some QACs reach ecotoxicological effect levels. This highlights the need to consider particle-associated QAC concentrations, the widespread use of QAC, as well as the environmental implications of chemical public health interventions.

Development of a QuEChERS-based method for the determination of quaternary ammonium compounds in different food matrices by LC-MS/MS

A QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) - based method was developed and validated for the determination of six benzalkonium chloride (BAC) homologues (C8-C18) and four didecyldimethylammonium chloride (DDAC) homologues (C8-C14) in various food commodities. Quaternary ammonium compounds (QACs) are cationic surfactants and biocides commonly found in cleaning and disinfecting products widely used in industrial applications, households, agriculture, hospitals, and healthcare facilities and their usage has increased since the COVID-19 pandemic. The method was validated in terms of specificity, selectivity, linearity, limit of quantification (LOQ), accuracy (recovery percentage), precision (CV %) and matrix effect (ME %). %). Intraday and interday precision were evaluated at two levels. Mean recoveries for all matrices fell within 70–120 %, with CV% below 20 %. All calibration curves demonstrated excellent linearity (R² ≥ 0.998). The LOQ was set and verified at 0.01 mg/kg for each analyte. The results were satisfactory according to the SANTE/2021/11312 guidelines. In addition, the method included the elimination of water at each stage of sample preparation and throughout the chromatographic system to minimize background noise. The method was successfully applied to analyse 46 samples of animal origin. BAC C14 was the most frequently detected compound (76 % of the samples) and in two samples, the sum of BACs was above the MRL set by EU the Regulation. These findings, coupled with limited data on QAC levels in meat and seafood products, underscore the necessity for further investigation into these matrices.

Adaptations to cationic biocide exposure differentially influence virulence factors and pathogenicity in Pseudomonas aeruginosa

ABSTRACT Cationic biocides (CBs), which include quaternary ammonium compounds (QACs), are employed to mitigate the spread of infectious bacteria, but resistance to such surface disinfectants is rising. CB exposure can have profound phenotypic implications that extend beyond allowing microorganisms to persist on surfaces. Pseudomonas aeruginosa is a deadly bacterial pathogen that is intrinsically tolerant to a wide variety of antimicrobials and is commonly spread in healthcare settings. In this study, we pursued resistance selection assays to the QAC benzalkonium chloride and quaternary phosphonium compound P6P–10,10 to assess the phenotypic effects of CB exposure in P. aeruginosa PAO1 and four genetically diverse, drug-resistant clinical isolates. In particular, we sought to examine how CB exposure affects defensive strategies and the virulence-associated “offensive” strategies in P. aeruginosa. We demonstrated that development of resistance to BAC is associated with increased production of virulence-associated pigments and alginate as well as pellicle formation. In an in vivo infection model, CB-resistant PAO1 exhibited a decreased level of virulence compared to wild type, potentially due to an observed fitness cost in these strains. Taken together, these results illustrate the significant consequence CB resistance exerts on the virulence-associated phenotypes of P. aeruginosa.

Cationic Biocides Tend to Embed into the Inner Layer of the Model Outer Membrane Vesicles of Gram-negative Bacteria: Computational Insights

The outer membrane vesicles (OMVs), produced by many pathogenic bacteria, play a significant role in bacterial pathogenesis. They promote bacterial resistance to antibiotics and act as natural protective barriers. The study of OMVs is essential both for understanding the general mechanisms of bacterial pathogenicity and for the development of the antibacterial drugs. In this paper, we created model vesicle that imitate the OMVs of Gram-negative bacteria using molecular modeling techniques. To investigate the interaction of the cationic antimicrobial compounds with the outer lipopolysaccharide (LPS) monolayer and the inner phospholipid monolayer of the OMV membrane, we performed molecular dynamics simulations by placing molecules of the cationic antiseptic octenidine on the outside or inside of model vesicles. The interaction of octenidine with the outer and inner monolayer was significantly different: octenidine interacted weakly with the outer LPS surface of the model OMV, but exhibited high affinity for the phospholipids of the inner monolayer. To study the translocation of cationic antimicrobial molecules within model OMV, we performed steered molecular dynamics simulations. For all three cationic biocide molecules, antiseptic octenidine, photosensitizer octakis(cholinyl)zinc phthalocyanine, and dye methylene blue, it turned out that, along with the LPS of the outer membrane of the OMV, the phosphates of lipid A molecules represent the final barrier to their penetration into the model OMV.

Current insights into the effects of cationic biocides exposure on Enterococcus spp.

Cationic biocides (CBs), such as quaternary ammonium compounds and biguanides, are critical for controlling the spread of bacterial pathogens like Enterococcus spp., a leading cause of multidrug-resistant healthcare-associated infections. The widespread use of CBs in recent decades has prompted concerns about the potential emergence of Enterococcus spp. populations exhibiting resistance to both biocides and antibiotics. Such concerns arise from their frequent exposure to subinhibitory concentrations of CBs in clinical, food chain and diverse environmental settings. This comprehensive narrative review aimed to explore the complexity of the Enterococcus' response to CBs and of their possible evolution toward resistance. To that end, CBs' activity against diverse Enterococcus spp. collections, the prevalence and roles of genes associated with decreased susceptibility to CBs, and the potential for co- and cross-resistance between CBs and antibiotics are reviewed. Significant methodological and knowledge gaps are identified, highlighting areas that future studies should address to enhance our comprehension of the impact of exposure to CBs on Enterococcus spp. populations' epidemiology. This knowledge is essential for developing effective One Health strategies that ensure the continued efficacy of these critical agents in safeguarding Public Health.

Chimeric Amphiphilic Disinfectants: Quaternary Ammonium/Quaternary Phosphonium Hybrid Structures.

Cationic biocides play a crucial role in the disinfection of domestic and healthcare surfaces. Due to the rise of bacterial resistance towards common cationic disinfectants like quaternary ammonium compounds (QACs), the development of novel actives is necessary for effective infection prevention and control. Toward this end, a series of 15 chimeric biscationic amphiphilic compounds, bearing both ammonium and phosphonium residues, were prepared to probe the structure and efficacy of mixed cationic ammonium-phosphonium structures. Compounds were obtained in two steps and good yields, with straightforward and chromatography-free purifications. Antibacterial activity evaluation of these compounds against a panel of seven bacterial strains, including two MRSA strains as well as opportunistic pathogen A. baumannii, were encouraging, as low micromolar inhibitory activity was observed for multiple structures. Alkyl chain length on the ammonium group was, as expected, a major determinant of bioactivity. In addition, high therapeutic indexes (up to 125-fold) for triphenyl phosphonium-bearing amphiphiles were observed when comparing antimicrobial activity to mammalian cell lysis activity.

Cleavable Amphiphilic Biocides with Ester-Bearing Moieties: Aggregation Properties and Antibacterial Activity.

The rise of multidrug-resistant bacterial infections and the dwindling supply of newly approved antibiotics have emerged as a grave threat to public health. Toward the ever-growing necessity of the development of novel antimicrobial agents, herein, we synthesized a series of cationic amphiphilic biocides featuring two cationic headgroups separated by different hydrophobic spacers, accompanied by the inclusion of two lipophilic tails through cleavable ester functionality. The detailed aggregation properties offered by these biocides were investigated by small-angle neutron scattering (SANS) and conductivity. The critical micellar concentration of the biocides and the size and shape of the micellar aggregates differed with variation of pendant and spacer hydrophobicity. Furthermore, the aggregation number and size of the micelles were found to vary with changing concentration and temperature. These easily synthesized biocides exhibited potent antibacterial properties against various multidrug-resistant bacteria. The optimized biocides with minimum hematotoxicity and potent antibacterial activity against methicillin-resistant Staphylococcus aureus and Acinetobacter baumannii exhibited rapid killing kinetics against planktonic bacteria. Also, these membrane-active agents were able to eradicate preformed biofilms. The enzymatic and acidic degradation profile further offered proof of gradual degradation. Collectively, these cleavable amphiphilic biocides demonstrated excellent potency for combating the multidrug-resistant bacterial infection.

Preparation and characterization of new antifouling coating based on alkyd paint modified with hydrophobic cationic biocide

Preparation and characterization of new antifouling coating based on alkyd paint modified with hydrophobic cationic biocide

Zooperiphyton on Anti-Fouling Coatings and Changes of Its Coenotic Structure

The features of the zooperiphyton communities' forming on experimental substrates - protective coatings with antifouling properties, based on industrial alkyd enamel PF-167 modified by cationic biocides 1-dodecylpyridine oleate and 1-dodecylpyridinium bis(2-ethylhexyl) phosphate in the bay of the Kaniv reservoir (the Dnieper river) have been studied. Some features of successional changes in the Zebra mussel and Bryozoans' (pearlwort) communities were discussed. On the contrast to the previous years, when the water temperature in August (time of high development of bryozoan colonies) was lower, the continual succession of communities with <i>Dreissena'</i>s dominance was not interrupted by the bryozoan's development. The protective coatings containing 8% of cationic biocides demonstrated high antifouling activity. In this study, no connections between the cenotic structure and nature of antifouling coatings were found.

Impacts of non-microbial soils on polychloramide disinfectants

The presence of some nonmicrobial chemicals and surfaces, herein called “soils”, are known to degrade the performance of biocides, and biocidal assays often include mixtures of materials to mimic the effects of soils. We hypothesized that water-soluble anionic polychloramide biocides were less sensitive to soil interference than cationic polymeric biocides. The relationships between soil composition and antimicrobial polymer biocidal activity were compared for an anionic polychloramide, a cationic polychloramide, and a cationic poly(quaternary ammonium) biocide. The nanoscale soil models individually investigated were polyacrylic acid (PAA), cellulose nanocrystals (CNCs), and bovine serum albumin. The low molecular weight model soils were ammonium chloride, glycine, and succinimide. Three types of soil impacts were identified: 1) sequestration, whereby the soil physically inhibited transport of the biocide to microbes; 2) extraction, whereby the soil reduced or extracted oxidative chlorine, decreasing or eliminating the oxidative chlorine strength; and 3) extraction whereby the biocidal activity increases in the presence of a low molecular weight chemical that carries oxidative Cl from the polymer to the microbes. PAA and CNCs inhibit cationic biocides by sequestration but have little impact on anionic polychloramide. Glycine and BSA extract oxidative chlorine, lowering the biocidal activity of the anionic and cationic polychloramides while not impacting the poly(quaternary ammonium) biocide. Finally, the presence of succinimide increased bacteria deactivation of both anionic and cationic polychloramides. We propose that succinimide extracts oxidative chlorine from the polychloramides and transports it to the bacteria.

Lipopolysaccharide structure modulates cationic biocide susceptibility and crystalline biofilm formation in Proteus mirabilis.

Chlorhexidine (CHD) is a cationic biocide used ubiquitously in healthcare settings. Proteus mirabilis, an important pathogen of the catheterized urinary tract, and isolates of this species are often described as "resistant" to CHD-containing products used for catheter infection control. To identify the mechanisms underlying reduced CHD susceptibility in P. mirabilis, we subjected the CHD tolerant clinical isolate RS47 to random transposon mutagenesis and screened for mutants with reduced CHD minimum inhibitory concentrations (MICs). One mutant recovered from these screens (designated RS47-2) exhibited ~ 8-fold reduction in CHD MIC. Complete genome sequencing of RS47-2 showed a single mini-Tn5 insert in the waaC gene involved in lipopolysaccharide (LPS) inner core biosynthesis. Phenotypic screening of RS47-2 revealed a significant increase in cell surface hydrophobicity and serum susceptibility compared to the wildtype, and confirmed defects in LPS production congruent with waaC inactivation. Disruption of waaC was also associated with increased susceptibility to a range of other cationic biocides but did not affect susceptibility to antibiotics tested. Complementation studies showed that repression of smvA efflux activity in RS47-2 further increased susceptibility to CHD and other cationic biocides, reducing CHD MICs to values comparable with the most CHD susceptible isolates characterized. The formation of crystalline biofilms and blockage of urethral catheters was also significantly attenuated in RS47-2. Taken together, these data show that aspects of LPS structure and upregulation of the smvA efflux system function in synergy to modulate susceptibility to CHD and other cationic biocides, and that LPS structure is also an important factor in P. mirabilis crystalline biofilm formation.

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 β-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.

Growth Inhibition of Sulfate-Reducing Bacteria during Gas and Oil Production Using Novel Schiff Base Diquaternary Biocides: Synthesis, Antimicrobial, and Toxicological Assessment.

Upstream crude oil production equipment is always exposed to destruction damagingly which is caused by sulfate-reducing bacterium (SRB) activities that produce H2S gas, which leads to increased metal corrosion (bio-fouling) rates and inflicts effective infrastructure damage. Hence, oil and gas reservoirs must be injected with biocides and inhibitors which still offer the foremost protection against harmful microbial activity. However, because of the economic and environmental risks associated with biocides, the oil and gas sectors improve better methods for their usage. This work describes the synthesis and evaluation of the biological activities as the cytotoxicity and antimicrobial properties of a series of diquaternary cationic biocides that were studied during the inhibition of microbial biofilms. The prepared diquaternary compound was synthesized by coupling vanillin and 4-aminoantipyrene to achieve the corresponding Schiff base, followed by a quaternization reaction using 1,6-bromohexane, 1,8-bromooctane, and 1,12-bromododecane. The increase of their alkyl chain length from 6 to 12 methylene groups increased the obtained antimicrobial activity and cytotoxicity. Antimicrobial efficacies of Q1-3 against various biofilm-forming microorganisms, including bacteria and fungi, were examined utilizing the diameter of inhibition zone procedures. The results revealed that cytotoxic efficacies of Q1-3 were significantly associated mainly with maximum surface excess and interfacial characteristics. The cytotoxic efficiencies of Q1-3 biocides demonstrated promising results due to their comparatively higher efficacies against SRB. Q3 exhibited the highest cytotoxic biocide against the gram +ve, gram -ve, and SRB species according to the inhibition zone diameter test. The toxicity of the studied microorganisms depended on the nature and type of the target microorganism and the hydrophobicity of the biocide molecules. Cytotoxicity assessment and antimicrobial activity displayed increased activity by the increase in their alkyl chain length.

Co-culture with Acinetobacter johnsonii enhances benzalkonium chloride resistance in Salmonella enterica via triggering lipid A modifications

Salmonella enterica is one of the leading causes of foodborne gastroenteritis worldwide. In the food production environment, many bacterial species co-exist on surfaces in biofilm structures, which can act as reservoirs of microbial contamination of food products. Polymicrobial biofilms have been shown to have greater tolerance to antimicrobials, such as disinfectants, however the mechanistic basis of this is poorly understood. In this study, S. enterica subsp. enterica serovar Liverpool was co-cultured in mixed-species biofilms with bacteria isolated from the food production environment and challenged with the cationic biocide disinfectant, benzalkonium chloride (BC). Co-culture with the common environmental bacterium Acinetobacter johnsonii resulted in >200-fold higher resistance of S. Liverpool to BC, compared to mono-culture biofilms. The transcriptional response of S. enterica to biofilm co-culture was determined using a dual RNA-seq strategy. Genes controlled by the PhoPQ and PmrAB two-component systems, involved in lipid A modification and associated with cationic antimicrobial peptide resistance (CAMP) of S. Liverpool, were significantly upregulated. Deletion of either the phoP or pmrA genes resulted in an increase in susceptibility to BC, suggesting that activation of their regulons during co-culture enhances BC resistance. S. Liverpool lipid A profiles changed significantly upon co-culturing, with greater incorporation of both phosphoethanolamine and palmitate, which was dependent upon activation of PhoPQ and PmrAB. We conclude that when grown in the presence of A. johnsonii, S. Liverpool increases its tolerance to cationic BC disinfection by remodelling its cell envelope including reducing the net negative charge of lipid A and increasing lipid A acyl density.

In-Membrane Nanostructuring of Cationic Amphiphiles Affects Their Antimicrobial Efficacy and Cytotoxicity: A Comparison Study between a De Novo Antimicrobial Lipopeptide and Traditional Biocides.

Cationic biocides have been widely used as active ingredients in personal care and healthcare products for infection control and wound treatment for a long time, but there are concerns over their cytotoxicity and antimicrobial resistance. Designed lipopeptides are potential candidates for alleviating these issues because of their mildness to mammalian host cells and their high efficacy against pathogenic microbial membranes. In this study, antimicrobial and cytotoxic properties of a de novo designed lipopeptide, CH3(CH2)12CO-Lys-Lys-Gly-Gly-Ile-Ile-NH2 (C14KKGGII), were assessed against that of two traditional cationic biocides CnTAB (n = 12 and 14), with different critical aggregation concentrations (CACs). C14KKGGII was shown to be more potent against both bacteria and fungi but milder to fibroblast host cells than the two biocides. Biophysical measurements mimicking the main features of microbial and host cell membranes were obtained for both lipid monolayer models using neutron reflection and small unilamellar vesicles (SUVs) using fluorescein leakage and zeta potential changes. The results revealed selective binding to anionic lipid membranes from the lipopeptide and in-membrane nanostructuring that is distinctly different from the co-assembly of the conventional CnTAB. Furthermore, CnTAB binding to the model membranes showed low selectivity, and its high cytotoxicity could be attributed to both membrane lysis and chemical toxicity. This work demonstrates the advantages of the lipopeptides and their potential for further development toward clinical application.

Disruption of Membrane Integrity as a Molecular Initiating Event Determines the Toxicity of Polyhexamethylene Guanidine Phosphate Depending on the Routes of Exposure.

Polyhexamethylene guanidine phosphate (PHMG-P), a cationic biocide, is widely used in household products due to its strong bactericidal activity and low toxicity. However, it causes fatal lung damage when inhaled. In this study, we investigated why PHMG-P causes fatal lung injury when inhaled, and demonstrated that the disruption of membrane integrity through ionic interaction—a molecular initiating event of PHMG-P—determines toxicity. Mice were injected intravenously with 0.9 or 7.2 mg/kg PHMG-P (IV group), or instilled intratracheally with 0.9 mg/kg PHMG-P (ITI group); they were euthanatized at 4 h and on days 1 and 7 after treatment. Increased total BAL cell count and proinflammatory cytokine production, along with fibrotic changes in the lungs, were detected in the ITI group only. Levels of hepatic enzymes and hepatic serum amyloid A mRNA expression were markedly upregulated in the 7.2 mg/kg IV and ITI groups at 4 h or day 1 after treatment, but returned to baseline. No pathological findings were detected in the heart, liver, or kidneys. To simulate the IV injection, A549, THP-1, and HepG2 cells were treated with PHMG-P in cell culture media supplemented with different serum concentrations. Increased serum concentration was associated with an increase in cell viability. These results support the idea that direct contact between PHMG-P and cell membranes is necessary for PHMG-induced toxicity.

Adsorption of cationic surfactant as a probe of the montmorillonite surface reactivity in the alginate hydrogel composites.

Adsorption of a cationic surfactant allowed to probe the surface reactivity of montmorillonite encapsulated in a composite of alginate hydrogels (A-MMT). Dodecylbenzyldimethylammonium chloride (BAC-12) was the surfactant used for these studies. BAC-12 is part of the widely used surfactant mixture known as benzalkonium chloride. XRD showed that up to three different types of basal spacing (d 001) were present within the composite indicating that as the concentration of adsorbed BAC-12 increases, populations with different adsorption conformational arrangements are present, even unexpanded clay remains. From the SEM-EDS spectra it is observed that the clay is distributed in the whole composite. In addition, the effect of the presence of cationic and anionic biocides on BAC-12 adsorption was studied. Cationic biocides such as tetradecyllbenzyldimethylammonium chlorides (BAC-14) and paraquat (PQ) show a competitive behavior for the clay adsorption sites at BAC-12 low concentration indicating an electrostatic adsorption mechanism. However, the presence of anionic contaminants such as 2,4-D and metsulfuron methyl do not affect surfactant adsorption. In all scenarios is observed an abrupt increase of BAC-12 adsorbed amount reaching values higher than the clay CEC suggesting strong tail-tail interactions. This occurs at concentrations 10 times lower than the CMC of BAC-12 promoted by clay encapsulation in the composite. In these composites the alginate does not affect the surface reactivity of the clay, but the formation of the hydrogel allows it to be easily extracted from aqueous media which makes it an interesting material with a potential use in water remediation.

Assessing the risk of resistance to cationic biocides incorporating realism-based and biophysical approaches.

The control of microorganisms is a key objective in disease prevention and in medical, industrial, domestic, and food-production environments. Whilst the effectiveness of biocides in these contexts is well-evidenced, debate continues about the resistance risks associated with their use. This has driven an increased regulatory burden, which in turn could result in a reduction of both the deployment of current biocides and the development of new compounds and formulas. Efforts to balance risk and benefit are therefore of critical importance and should be underpinned by realistic methods and a multi-disciplinary approach, and through objective and critical analyses of the literature. The current literature on this topic can be difficult to navigate. Much of the evidence for potential issues of resistance generation by biocides is based on either correlation analysis of isolated bacteria, where reports of treatment failure are generally uncommon, or laboratory studies that do not necessarily represent real biocide applications. This is complicated by inconsistencies in the definition of the term resistance. Similar uncertainties also apply to cross-resistance between biocides and antibiotics. Risk assessment studies that can better inform practice are required. The resulting knowledge can be utilised by multiple stakeholders including those tasked with new product development, regulatory authorities, clinical practitioners, and the public. This review considers current evidence for resistance and cross-resistance and outlines efforts to increase realism in risk assessment. This is done in the background of the discussion of the mode of application of biocides and the demonstrable benefits as well as the potential risks.