Antibiotic resistance is a major public health concern. Triclosan is an antimicrobial compound with direct links to antibiotic resistance that was widely used in soaps in the U.S. until its ban by the U.S. Food and Drug Administration. Benzalkonium chloride (BAC), a quaternary ammonium compound, has widely replaced triclosan in soaps marketed as an antibacterial. BAC has been detected in surface waters and its presence will likely increase following increased use in soap products. The objective of this study was to determine the effect of BAC on relative abundance of antibiotic resistance in a bacterial community from a surface water used as a source for drinking water treatment. Bench-scale microcosm experiments were conducted with microbial communities amended with BAC at concentrations ranging from 0.1 μg L⁻¹ to 500 μg L⁻¹. Phenotypic antibiotic resistance was quantified by culturing bacteria in the presence of different antibiotics, and genotypic resistance was determined using qPCR to quantify antibiotic resistance genes (ARGs). BAC at concentrations ranging from 0.1 μg L⁻¹ to 500 μg L⁻¹ was found to positively select for bacteria resistant to ciprofloxacin and sulfamethoxazole, and negatively select against bacteria with resistance to six other antibiotics. Exposure to BAC for 14 days increased the relative abundance of sul1 and blaTEM. This study re-highlights the importance of employing both culture and non-culture-based techniques to identify selection for antibiotic resistance. The widespread use of BAC will likely impact antibiotic resistance profiles of bacteria in the environment, including in source waters used for drinking water, wastewater treatment plants, and natural waterways.

Benzylalkyldimethylethyl ammonium compounds are pervasive in natural environments and toxic at high concentrations. The changes in functional genes and microbial diversity in eutrophic lake samples exposed to benzyldimethyldodecyl ammonium chloride (BAC) were assessed. BAC exerted negative effects on bacteria abundance, particularly at concentrations of 100 μg L−1 and higher. A significant increase in the number of the quaternary ammonium compound-resistant gene qacA/B was recorded within the 10 μg L−1 treatment after the first day of exposure. Not all antibiotic resistance genes increased in abundance as the concentrations of BAC increased; rather, gene abundances were dependent on the gene type, concentrations of BAC, and contact time. The nitrogen fixation-related gene nifH and ammonia monooxygenase gene amoA were inhibited by high concentrations of BAC after the first day, whereas an increase of the nitrite reductase gene nirK was stimulated by exposure. Microbial communities within higher treatment levels (1000 and 10 000 μg L−1) exhibited significantly different community composition compared to other treatment levels and the control. Selective enrichment of Rheinheimera, Pseudomonas, and Vogesella were found in the higher treatment levels, suggesting that these bacteria have some resistance or degradation capacity to BAC. Genes related with RNA processing and modification, transcription, lipid transport and metabolism, amino acid transport and metabolism, and cell motility of microbial community function were involved in the process exposed to the BAC stress.

With the recent ban of triclosan (TCS) and triclocarban (TCC) from some personal care products, many replacement antimicrobial compounds have been used. Yet the potential health risk and environmental impact of these replacement compounds are largely unknown. Here we investigated the toxicological effects of three commonly used replacement antimicrobials, benzalkonium chloride (BAC), benzethonium chloride (BEC), and chloroxylenol (CX) to two model organisms, the nematode C. elegans and zebrafish (Danio rerio), and compared them to the banned TCS and TCC. We found that these replacement compounds are not any safer than the banned antimicrobials. In the worm, at least one of the three, BAC, showed comparable toxicity to TCS from organismal to molecular levels, with toxic effects occurring at lower hundred μg/L to lower mg/L levels. In the fish, all three compounds at the tested concentration ranges (0.05–5 mg/L) showed toxicity effects to zebrafish embryos, indicated by hatching delay or inhibition, embryonic mortality, morphological malformations, and neurotoxicity. BAC was the most toxic among the three, with acute lethal toxicity occurring at environmentally relevant concentrations (hundreds of μg/L), which is comparable to the banned TCC. However, the toxicity effects of BAC and TCC occurred within different time windows, potentially suggesting different mechanisms of toxicity. CX was the only compound that induced a “body curvature” phenotype among the five compounds examined, suggesting a unique mode of toxic action for this compound. Furthermore, all five compounds except TCS induced neurotoxicity in fish larvae, indicated by alterations in secondary motoneuron axonal projections. Such neurotoxicity has been largely understudied for these antimicrobials in the past years and calls for further investigations in terms of its underlying mechanisms and ecological significance. These findings strongly indicate that scrutiny should be put on these replacement compounds before their introduction into massive use in personal care products.

The toxicity and genotoxicity of the cationic surfactant benzalkonium chloride (BAC) were studied using Daphnia magna and Ceriodaphnia dubia as model systems. Acute and chronic toxicity testing were performed according to the international standard guidelines and the genotoxicity was detected through the comet assay on cells from whole organisms in vivo exposed. Acute effects occurred at concentrations in the order of tens of μg/L in D. magna and hundreds of μg/L in C. dubia. Chronic effects were found at one order of magnitude less than short-term effects maintaining the same difference in sensitivity between D. magna and C. dubia. BAC induced relevant DNA damage, in both cladocerans; the lowest adverse effect levels were 0.4 and 4 ng/L for D. magna and C. dubia, respectively. As these effective concentrations are far lower than BAC occurrence in surface waters (units of μg/L) a concerning environmental risk cannot be excluded. The findings of this study showed that D. magna and C. dubia, could be used as model organisms to detect acute and chronic toxicity as well as genotoxicity at the whole organism level.

The increased use of disinfectants due to the spread of the novel coronavirus infection (e.g. COVID-19) has caused burden in the environment but knowledge on its ecotoxicological impact on the estuary environment is limited. Here we report in vivo and molecular endpoints that we used to assess the effects of chloroxylenol (PCMX) and benzalkonium chloride (BAC), which are ingredients in liquid handwash, dish soap products, and sanitizers used by consumers and healthcare workers on the estuarine rotifer Brachionus koreanus. PCMX and BAC significantly affected the life table parameters of B. koreanus. These chemicals modulated the activities of antioxidant enzymes such as superoxide dismutase and catalase and increased reactive oxygen species even at low concentrations. Also, PCMX and BAC caused alterations in the swimming speed and rotation rate of B. koreanus. Furthermore, an RNA-seq-based ingenuity pathway analysis showed that PCMX affected several signaling pathways, allowing us to predict that a low concentration of PCMX will have deleterious effects on B. koreanus. The neurotoxic and mitochondrial dysfunction event scenario induced by PCMX reflects the underlying molecular mechanisms by which PCMX produces outcomes deleterious to aquatic organisms.

In this work, the distribution of quaternary ammonium compounds (QACs) in two dated sediment cores, collected from the Pearl River Estuary (PRE) and Tokyo Bay (TB), were investigated to understand the historical input of QACs and their diagenetic behavior in urban estuarine environments. The vertical variation profiles of QAC concentrations showed that benzylalkyldimethyl ammonium compounds (BACs) and dialkyldimethyl ammonium compounds (DADMACs) were widely used during 1970s and 1980s both in China and Japan. The declining environmental concentrations of QACs suggested a compositional change of commodities and the effectiveness of emission control strategies. For the individual QAC homologues, BAC homologues decreased significantly over time, while DADMAC compositions remained relatively stable. The differences in concentration and composition profiles of BACs and DADMACs in the sediment cores provided useful information on the patterns of use of QACs in China and Japan, as well as their diagenetic behaviors in the sediments.

A previously isolated indigenous strain of Pseudomonas sp. was used to treat effluents, a synthetic and an industrial-containing benzalkonium chloride (BAC), in continuous upflow biofilm aerobic reactors. The reactor used to treat the synthetic effluent was constructed from Plexiglas® and filled with hollow polyvinyl chloride (PVC) cylinders as support material, whereas the one used to treat the industrial effluent was constructed from PVC and had a high recirculation flow rate and lightweight expanded clay aggregate (LECA®) as support material. Biodegradation was evaluated by spectrophotometry, HPLC, and microbial growth. Detoxification was evaluated by using Vibrio fisheri, Pseudokirchneriella subcapitata, and Lactuca sativa as test organisms. Maximal BAC influent concentrations were of 383.4 and 1172.0 mg L⁻¹ respectively, which corresponds to a maximal organic load of 49.8 and 146.5 g BAC m⁻³ day⁻¹. The efficiency of the reactors was higher than 99.3% in terms of compound removal and 97.0% in terms of COD removal. Complete detoxification of the effluent was demonstrated for the synthetic effluent, whereas a toxicity removal higher than 97% was reached in the case of the industrial effluent. The promising behavior of the isolated indigenous strain to degrade BAC in continuous reactors allows us to suggest its possible use in remediation processes.

Benzalkonium chloride (BAC) loaded to montmorillonites (Mt) or organomontmorillonites (OMt) generates a functional material that can be incorporated to several systems (polymers, paints, etc) as a controlled release bactericide. Understanding the BAC adsorption sites on these adsorbents is of high importance to clarify their adsorption/desorption characteristics in aqueous media or other solvents. In this work, a thorough study about the adsorption/desorption properties of Mt and OMt with regards to BAC is presented, in order to evaluate further BAC release with the consequent aquatic environment contamination. In this work, the BAC adsorption on two different sites is demonstrated: the interlayer space and the external surface. Depending on BAC concentration in water, sorption of BAC at Mt occurred in two steps. At adsorbed amount <0.5 mmol g⁻¹, there was an Mt interlayer expansion of 0.49 nm with no change of the external charge. At adsorbed amount >0.5 mmol g⁻¹, there was a new interlayer expansion attaining 0.75 nm and the external charge shifted to positive value. In the case of OMt, the introduction of BAC produced changes in the interlayer structure and in the external surface charge. BAC desorption was strongly dependent on the type of Mt or OMt and extraction solvent, knowledge of which will allow its safe use in environmental friendly technological applications.

A combined use of silver nanoparticles (AgNPs) with different types of disinfectants as antimicrobial might be useful in mitigating the problem of development of bacterial resistance with a strong enhancement of the biocidal effect of disinfectants. To evaluate the biocidal activity of silver nanoparticles and its loaded forms, five commercial disinfectants (quaternary ammonium compounds (benzalkonium chloride (BC) and TH⁴⁺), Virkon®S, sodium hypochlorite, and hydrogen peroxide (H₂O₂)) were used against Listeria monocytogenes (L. monocytogenes) isolates at different concentrations and exposure times to reveal intra-species variability and the percentage of resistance to antimicrobial agents used. Therefore, a total of 260 specimens from animal and human stool as well as environmental samples from dairy cattle farms were cultured for isolation of L. monocytogenes. Thereafter, bacterial isolates were identified using PCR. Silver nanoparticle was synthesized using chemical reduction. Both silver nanoparticles and its loaded forms were characterized by transmission electron microscopy (TEM). The sensitivity test of 60 strains of L. monocytogenes bacteria to AgNPs and its loaded forms was evaluated using broth macrodilution method. Virkon®S/AgNPs 2.0% exhibited the highest bactericidal effect (100%) against L. monocytogenes strains followed by H₂O₂/AgNPs 5.0% and TH⁴⁺/AgNPs 1.0% (90% each). Furthermore, the percentage of resistance of L. monocytogenes was 0.0% to both H₂O₂/AgNPs 5.0% and Virkon®S/AgNPs 2.0%. In conclusion, monitoring the main source of contamination with Listeria monocytogenes in dairy cattle farms is an essential factor to achieve an efficient control. Moreover, the use of the disinfectants, Virkon®S 2.0%, H₂O₂ 5.0%, and TH⁴⁺1.0%, loaded on silver nanoparticles composite had the strong bactericidal effect against L. monocytogenes.

The release of anthropogenic compounds into the aquatic environment has been a particular concern, since some of these substances exhibit biologic activity of different types in non-target species. Among anthropogenic compounds present in the aquatic compartment, detergents are commonly found and may be responsible for physiological modifications in exposed organisms. The impairment of key physiological functions, such as neurotransmission, and tissue damage in some important organs, has been used to assess the effects of several classes of xenobiotics, including detergents, in aquatic organisms. The present study intended to assess the effect of three types of detersive compounds (sodium dodecylsulfate (SDS), benzalkonium chloride (BZC), and Triton X-100 (TX100)) in the acetylcholinesterase activity (AChE) and tissue damage (gills and liver) of Gambusia holbrooki after a chronic exposure to realistic levels of these compounds. SDS, BZC, and TX100 did not cause any significant alteration in AChE. Furthermore, no specific gross morphological changes were also observed in the gills and liver of the exposed individuals. It is possible to conclude that, under ecologically relevant conditions of exposure, both tissue damage and cholinesterasic impairment are not toxicological pathways affected by detergents in G. holbrooki.