Common Biocides Research Articles

Sensitivity of 240 Listeria monocytogenes isolates to common industrial biocides is more dependent on the presence of residual organic matter or biofilm than on genetic determinants

Listeria monocytogenes continues to be a problem in ready-to-eat foods. Tolerance and adaptation to biocides have been proposed to contribute to the persistence of L. monocytogenes in the food processing industry. This study aimed to determine if the sensitivity to common biocides varies among 240 L. monocytogenes isolates. The impact of residual organic matter (soils), biofilm, and ability to adapt to biocides was also evaluated. Food and food processing related isolates of L. monocytogenes (n = 240) were whole genome sequenced and assigned to sequence types (n = 53) and clonal complexes (n = 32). Isolates were screened for genetic elements associated with biocide tolerance and tested for their minimum inhibitory concentration (MIC) to benzalkonium chloride (BC), peracetic acid (PAA), sodium hypochlorite (SH) and ethanol (ET). Biocide tolerance under soiled conditions, in biofilm and in a broth suspension test (BST) was further tested for 19 representative isolates. Adaptation to sub-MICs of BC, PAA or SH (18 days, 15 °C) was tested for 15 isolates. MIC for PAA (62 mg/L) showed no variation, while for SH (47–94 mg/L) and ET (4.7–9.4% v/v) values differed 2 × among isolates. For BC, an 8 × difference (0.3–2.5 mg/L) was observed. Eighty-seven of 88 BC tolerant isolates (MIC of 2.5 mg/L) harbored known BC tolerance genes. Soiled conditions increased MIC-values 8–33 × for BC and 8 × for SH but not for PAA and ET. Biofilm increased minimum bactericidal concentrations 4–8 × for BC but not for PAA and SH. Survival of isolates with BC tolerance genes was not significantly (p > 0.05) improved in BSTs with PAA, SH or BC. Lineage I isolates (n = 7 of 19) showed significantly (p < 0.05) better survival than lineage II isolates (n = 12) when exposed to PAA (100 mg/L) in BSTs. Adaptation to BC (8 × MIC increase) occurred exclusively for originally sensitive isolates, while no adaptation to SH and PAA were observed. In conclusion, L. monocytogenes isolates showed limited variation in sensitivity to biocides in MIC assays. However, lineage I isolates were less sensitive to PAA than lineage II isolates in BSTs. The efficacy of SH and BC were affected by residual organic matter and/or biofilm, demonstrating the need for proper cleaning prior to biocide use with PAA being less perturbed.

Identification of norA, qacAB and smr genes and determination of antibiotic and biocide resistance patterns in methicillin-resistant Staphylococcus aureus isolated from Isfahan-Iran

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of nosocomial infections, which have become a serious problem due to antibiotic resistance. In addition to antibiotic resistance, biocide resistance is also increasing due to the high use of these substances. The aim of this study was to investigate the pattern of antibiotic resistance and the prevalence of biocide resistance genes norA, qacA/B and smr, and to determine the minimum inhibitory concentration of chlorhexidine digluconate (CHG) and benzalkonium chloride (BKC) in MRSA isolates taken from teaching hospitals in Isfahan city, Iran. Materials and Methods: This study was conducted on 63 MRSA isolates collected from teaching hospitals in Isfahan city, Iran. MRSA isolates were identified by performing phenotypic testing and then confirmed by using PCR. The resistance pattern was determined by using disc diffusion method. Broth microdilution method was performed to determine the minimum inhibitory concentration (MIC) of CHG and BKC according to the Clinical and Laboratory Standards Institute (CLSI). The presence of biocide resistance genes was determined by using PCR and the Time kill assay was performed to investigate the lethality of biocides. Results: MRSA isolates had MIC values of 0.5–4 μg/ml for BKC and 0.125–4 μg/ml for CHG. norA was the most common biocide resistance gene among 63 isolates (88.9%), followed by qacAB and smr with 28.6% and 22.2%, respectively. Among the isolates, the highest percentage of resistance belonged to penicillin, erythromycin and doxycycline, respectively. Conclusion: Due to the high prevalence of MRSA isolates in hospitals and the role of this pathogen in nosocomial infections, continuous monitoring of antimicrobial resistance for appropriate antibiotic administration and continuous evaluation of biocide resistance due to the high prevalence of resistance genes and appropriate use of biocides for disinfection, allows us to minimize further prevalence of resistant strains in hospital environments.

Toxicity of common biocides used in aquaculture to embryos and larvae of Brachymystax tsinlingensis Li.

Brachymystax tsinlingensis Li is a threatened fish species endemic to China. With the problems of environmental factors and seeding breeding diseases, it is important to further improve the efficiency of seeding breeding and the basis of resource protection. This study investigated the acute toxicity of copper, zinc and methylene blue (MB) on hatching, survival, morphology, heart rate (HR) and stress behaviour of B. tsinlingensis. Eggs (diameter: 3.86 ± 0.07 mm, weight: 0.032 ± 0.004 g) of B. tsinlingensis were selected randomly from artificial propagation and developed from eye-pigmentation-stage embryos to yolk-sac stage larvae (length: 12.40 ± 0.02 mm, weight: 0.03 ± 0.001 g) and exposed to different concentrations of Cu, Zn and MB for 144 h in a series of semi-static toxicity tests. The acute toxicity tests indicated that the 96-h median lethal concentration (LC50 ) values of the embryos and larvae were 1.71 and 0.22 mg l-1 for copper and 2.57 and 2.72 mg l-1 for zinc, respectively, whereas the MB LC50 after 144-h exposure for embryos and larvae were 67.88 and 17.81 mg l-1 , respectively. The safe concentrations of copper, zinc and MB were 0.17, 0.77 and 6.79 mg l-1 for embryos and 0.03, 0.03 and 1.78 mg l-1 for larvae, respectively. Copper, zinc and MB treatments with concentrations greater than 1.60, 2.00 and 60.00 mg l-1 , respectively, led to a significantly low hatching rate and significantly high embryo mortality (P < 0.05), and copper and MB treatments with concentrations greater than 0.2 and 20 mg l-1 led to significantly high larvae mortality (P < 0.05). Exposure to copper, zinc and MB resulted in developmental defects, including spinal curvature, tail deformity, vascular system anomalies and discolouration. Moreover, copper exposure significantly reduced the HR of larvae (P < 0.05). The embryos exhibited an obvious change in behaviour, converting from the normal behaviour of emerging from the membrane head first to emerging tail first, with probabilities of 34.82%, 14.81% and 49.07% under copper, zinc and MB treatments, respectively. The results demonstrated that the sensitivity of yolk-sac larvae to copper and MB was significantly higher than that of embryos (P < 0.05) and that B. tsinlingensis embryos or larvae might be more resistant to copper, zinc and MB than other members of the Salmonidae family, which benefits their resource protection and restoration.

Effects of currently used marine antifouling paint biocides on green fluorescent proteins in Anemonia viridis.

Some of the antifouling booster biocides affects the marine ecosystem negatively. The booster biocides that are resistant to degradation are accumulated in the sediment of the oceans. One of the sedentary organisms in the Mediterranean Sea is Anemonia viridis. This study aims at showing the toxicities of common biocides such as irgarol, seanine-211, zinc omadine, and acticide on the fluorescence by GFPs of A. viridis. The decreases in the fluorescence intensities of the GFP were measured within different booster biocide concentrations. The results show that fluorescent intensities of GFP proteins decrease more than 50% when they are exposed to different concentrations of irgarol, zinc omadine, acticide. In conclusion, ecosystem health should be prioritized when new antifouling paint compositions are proposed. From the results, it seems that A. viridis can be considered as a vulnerable organism and it is sensitive to booster biocides within self-polishing antifouling paint formulations.

Antifouling coating based on biopolymers (PCL/ PLA) and bioactive extract from the sea cucumber Stichopus herrmanni

An important challenge to decrease the toxic effects of the common biocides in marine environments and to achieve suitable ecofriendly natural antifouling coatings is to find effective natural antifoulants and efficient biodegradable coatings. In this study, antifouling activities of nine bioactive extracts (non-polar to polar) from different organs of the sea cucumber Stichopus herrmanni were tested against five bacterial strains, barnacle and brine shrimp larvae. The ethyl acetate extract of the body wall showed the highest in-vitro antifouling activity including high antibacterial and anti-barnacle activities and low toxicity against the brine shrimp as non-target organism. Based on these results, 10 phr of the ethyl acetate extract from S.herrmanni was added to different coatings consisting of polycaprolactone (PCL)/polylactic acid (PLA) blends containing various compositions of PLA (0, 10, and 20 wt.%). Polyvinyl chloride panels were coated with the prepared antifouling coatings and immersed in seawater for three months. Panel coated with PCL 80% /PLA 20% containing 10 phr of the antifoulant (panel-5), showed the highest resistance against fouling settlement with fouling coverage of 41.66% (P < 0.05). In addition, the lowest fouling weight was measured in panel-5 as well (81.00 ± 9.85 g) (P < 0.05). These findings indicate the antibacterial and antifouling potential of semi-polar bioactive extracts from the S. herrmanni body wall as natural antifoulants, as well as the enhanced antifouling performance of PCL/the natural antifoulant coatings by adding PLA.Graphical

Peracetate/singlet oxygen chemistry used in post-bleaching of kraft pulp as a practical oxidant for paper machines

The use of a novel sodium peracetate/singlet oxygen chemistry for brightening bleached kraft pulp shows exciting potential for technical performance, supply logistics, safety, and cost reduction. Potential chemical carryover to the paper machine raises questions about whether peracetate will impact paper machine performance, such as metal corrosion, useful press felt life, and interference with existing biocide programs or paper machine chemistry. Sodium peracetate/singlet oxygen chemistry can be used in high-density storage chests for brightening/whitening and to increase color stability. Any oxidant used directly before the paper machine has the possibility of impacting paper machine operations. Traditional oxidants used in bleaching, such as chlorine dioxide and hydrogen peroxide, are known to cause corrosion on machinery metals and press felts. Hydrogen peroxide residuals can interfere with common biocide programs. Traditional oxidants used in biocide treatments themselves significantly degrade press felt life when the rule-of-thumb concentration thresholds are exceeded. Sodium peracetate is evaluated in this paper for its impact on nylon press felt fiber degradation, metal corrosion, and interference with typical biocide programs. Laboratory results indicate that sodium peracetate/singlet oxygen chemistry is less corrosive than chlorine, bro-mine, and hydrogen peroxide on press felt nylon fiber and can therefore be used at higher levels than those chemistries to increase brightness without increasing negative downstream impact. Sodium peracetate can also be used with current biocide programs without negative impacts such as consumptive degradation. Higher residuals of per-acetate going to the paper machine may be useful as a biocide itself and can complement existing programs, allowing those programs to stay within their safe operating levels and thereby extend press felt useful life.

Transcriptomic analysis of the food spoilers Pseudomonas fluorescens reveals the antibiofilm of carvacrol by interference with intracellular signaling processes

Pseudomonas fluorescens as an important food spoilage organism, has strong biofilm forming-ability to resist common biocides, increasing persistence and contamination in the cold fresh food chain. This bacterial biofilm organization is regulated by luxI/luxR mediated quorum sensing (QS) signaling. Carvacrol with antimicrobial properties is well documented, however, the mechanisms of its antibiofilm activity targeting QS are still not well known. The aim of this study was to explore the inhibition of carvacrol on biofilm formation and spoilage potential of two P. fluorescens PFuk4 and PF07 strains, and their luxI mutants (ΔluxI). The minimum inhibitory concentration (MIC) of carvacrol against planktonic wild type (WT) strains and the corresponding ΔluxI was 1.6 and 1.4 mM, respectively, and maximum sublethal concentration (MSC) was 0.8 mM. The acyl-homoserine-lactones (AHLs) C4-HSL and C6-HSL, as major autoinducer molecules, were greatly decreased in PFuk4 and PF07 treated with 0.4 mM carvacrol, while those in the ΔluxI were almost abolished. Biofilm formation and exopolysaccharide production of the P. fluorescens WT and ΔluxI strains were significantly inhibited by carvacrol at the MSC, as well as the observed sparer and thinner biofilm structures. Bacterial cell swimming and swarming motilities were both decreased by carvacrol without affecting cell viability. Protease activity secreted by P. fluorescens was significantly reduced treated by MSC of carvacrol, in consistent with the repression of sensory attributes, especially in the WT strains. Transcriptomics profiling revealed that exposure to 0.4 mM carvacrol dramatically altered gene expression in P. fluorescens PF07 biofilm, as a total of 402 differentially expressed genes (DEGs) were upregulated and 308 DEGs were downregulated, compared with the untreated biofilms. Those DECs were mainly involved in pathways of ribosome, bacterial chemotaxis, citrate cycle. Furthermore, carvacrol also altered the transcription of P. fluorescens genes associated with flagellar assembly, chemotaxis, exopolysaccharide synthesis, signaling pathways of LuxR mediated QS and c-di-GMP, and amino acid degradation, which may contribute to biofilm development and spoilage potential. In addition, carvacrol was found to competitively bind with LuxR predicted protein to interrupt the interaction with C4-HSL, supported by molecular docking. Our results provide novel insights into the regulatory signaling networks of antibiofilm formation in P. fluorescens affected by carvacrol.

Schiff-base silver nanocomplexes formation on natural biopolymer coated mesoporous silica contributed to the improved curative effect on infectious microbes

Infectious microbes that spread easily in healthcare facilities remain as the severe threat for the public health, especially among immunocompromised populations. Given the intricate problem of dramatic increase in resistance to common biocides, the development of safe and efficient biocide formulated agents to alleviate drug resistance is highly demanding. In this study, Schiff-base ligands were successfully formed on natural biopolymer of epsilon-poly-L-lysine (e-PL) decorated aldehyde functionalized mesoporous silica SBA-15 (CHO-SBA-15) for the selective coordination of silver ions, which was affirmed by various physicochemical methods. Besides the identified broad-spectrum antibacterial activities, the as-prepared Schiff-base silver nanocomplex (CHO-SBA-15/e-PL/Ag, CLA-1) exhibited an improved inhibitory effect on infectious pathogen growth typified by Escherichia coli and Staphylococcus aureus in comparison with two control silver complexes without Schiff-base conjugates, SBA-15/e-PL/Ag and CHO-SBA-15/Ag, respectively. In addition, CLA-1 remarkably inhibited the growth of Mycobacterium tuberculosis due to the excellent antimicrobial activity of silver species. Significantly, CLA-1 kills Candida albicans cells, inhibits biofilm formation, and eliminates preformed biofilms, with no development of resistance during continuous serial passaging. The antifungal activity is connected to disruption of bacterial cell membranes and increased levels of intracellular reactive oxygen species. In mouse models of multidrug-resistant C. albicans infection, CLA-1 exhibited efficient in vivo fungicidal efficacy superior to two antifungal drugs, amphotericin B and fluconazole. Moreover, CLA-1 treatment induces negligible toxicity against normal tissues with safety. Therefore, this study reveals the pivotal role of the molecular design of Schiff-base silver nanocomplex formation on biopolymer surface-functionalized silica mesopores as a green and efficient nanoplatform to tackle infectious microbes.

Antimicrobial Activity of Five Apitoxins from Apis mellifera on Two Common Foodborne Pathogens.

Antimicrobial resistance is one of today’s major public health challenges. Infections caused by multidrug-resistant bacteria have been responsible for an increasing number of deaths in recent decades. These resistant bacteria are also a concern in the food chain, as bacteria can resist common biocides used in the food industry and reach consumers. As a consequence, the search for alternatives to common antimicrobials by the scientific community has intensified. Substances obtained from nature have shown great potential as new sources of antimicrobial activity. The aim of this study was to evaluate the antimicrobial activity of five bee venoms, also called apitoxins, against two common foodborne pathogens. A total of 50 strains of the Gram-negative pathogen Salmonella enterica and 8 strains of the Gram-positive pathogen Listeria monocytogenes were tested. The results show that the minimum inhibitory concentration (MIC) values were highly influenced by the bacterial genus. The MIC values ranged from 256 to 1024 µg/mL in S. enterica and from 16 to 32 µg/mL in L. monocytogenes. The results of this study demonstrate that apitoxin is a potential alternative agent against common foodborne pathogens, and it can be included in the development of new models to inhibit the growth of pathogenic bacteria in the food chain.

Development of a highly efficient electrochemical flow-through anode based on inner in-site enhanced TiO2-nanotubes array

This paper reports on the development of macroporous flow-through anodes. The anodes comprised an enhanced TiO2 nanotube array (ENTA) that was grown on three macroporous titanium substrates (MP-Ti) with nominal pore sizes of 10, 20, and 50 µm. The ENTA was then covered with SnO2-Sb2O3. We refer to this anode as the MP-Ti-ENTA/SnO2-Sb2O3 anode. The morphology, pore structure, and electrochemical properties of the anode were characterized. Compared with the traditional NTA layer, we found that the MP-Ti-ENTA/SnO2-Sb2O3 anode has a service lifetime that was 1.56 times larger than that of MP-Ti-NTA/SnO2-Sb2O3. We used 2-methyl-4-isothiazolin-3-one (MIT), a common biocide, as the target pollutant. We evaluated the impact of the operating parameters on energy efficiency and the oxidation rate of MIT. Furthermore, the apparent rate constants were 0.38, 1.63, and 1.24 min−1 for the 10, 20, and 50 μm nominal pore sizes of the MP-Ti substrates, respectively, demonstrating the different coating–loading mechanisms for the porous substrate. We found that hydroxyl radicals were the dominant species in the MIT oxidation in the HO radical scavenging experiments. The radical and nonradical oxidation contributions to the MIT degradation for different current densities were quantitatively determined as 72.1%–74.8% and 25.2%–27.9%, respectively. Finally, we summarized the oxidation performance for MIT destruction for (1) the published literature on various advanced oxidation technologies, (2) the published literature on various anodes, and (3) our flow-by and -through anodes. Accordingly, we found that our flow-through anode has a much lower electrical efficiency per order value (0.58 kWh m−3) than the flow-by anodes (6.85 kWh m−3).

Food safety risk during the pandemic

Food safety risk during the pandemic

Biocidal activity of polylactic acid-based nano-formulated abamectin on Acyrthosiphon pisum (Hemiptera: Aphididae) and the aphid predator Adalia bipunctata (Coleoptera: Coccinellidae).

Abamectin is a common biocide used to control agricultural insect pests. However, the water insolubility of abamectin may result in extra organic solvent introduced in the environment. To solve this issue, it is desirable to develop nanoformulations to encapsulate abamectin with environment-friendly polymers. In this study, two polylactic acid based abamectin nanoformulations were prepared. The average particle sizes, measured by dynamic light scattering and transmission electron microscope, were 240 nm and 150 nm, respectively. The insecticidal activity of these nano-formulated abamectin was examined in the laboratory on the pea aphid, Acyrthosiphon pisum (Hemiptera: Aphididae). The acute toxicity of nano-formulated abamectin on non-target aphid predator Adalia bipunctata (Coleoptera: Coccinellidae) was also evaluated by topical, residual and oral exposure. The two nano-formulated abamectin had comparable insecticidal effect with commercial abamectin formulation against the pea aphid. Taking median lethal concentration (LC50) as the toxicological endpoint, nanoformulations had higher contact toxicity and lower oral toxicity to first-instar larvae of the predator A. bipunctata. These results are expected to contribute to the application of solvent-free nano-formulated pesticides that comply with the integrated pest management (IPM) strategies.

Backtracking and forward checking of human listeriosis clusters identified a multiclonal outbreak linked to Listeria monocytogenes in meat products of a single producer

ABSTRACT Due to its high case fatality rate, foodborne listeriosis is considered a major public health concern worldwide. We describe one of the largest listeriosis outbreaks in Germany with 83 cases of invasive listeriosis between 2013 and 2018. As part of the outbreak investigation, we identified a highly diverse Listeria monocytogenes population at a single producer of ready-to-eat meat products. Strikingly, the extensive sampling after identification of a first match between a cluster of clinical isolates and a food isolate allowed for a linkage between this producer and a second, previously unmatched cluster of clinical isolates. Bacterial persistence in the processing plant and indications of cross-contamination events explained long-term contamination of food that led to the protracted outbreak. Based on screening for virulence factors, a pathogenic phenotype could not be ruled out for other strains circulating in the plant, suggesting that the outbreak could have been even larger. As most isolates were sensitive to common biocides used in the plant, hard to clean niches in the production line may have played a major role in the consolidation of the contamination. Our study demonstrates how important it is to search for the origin of infection when cases of illness have occurred (backtracking), but also clearly highlights that it is equally important to check whether a contamination at food or production level has caused disease (forward checking). Only through this two-sided control strategy, foodborne disease outbreaks such as listeriosis can be minimized, which could be a real improvement for public health.

Combinations of Antimicrobial Polymers with Nanomaterials and Bioactives to Improve Biocidal Therapies.

The rise of antibiotic-resistant microorganisms has become a critical issue in recent years and has promoted substantial research efforts directed to the development of more effective antimicrobial therapies utilizing different bactericidal mechanisms to neutralize infectious diseases. Modern approaches employ at least two mixed bioactive agents to enhance bactericidal effects. However, the combinations of drugs may not always show a synergistic effect, and further, could also produce adverse effects or stimulate negative outcomes. Therefore, investigations providing insights into the effective utilization of combinations of biocidal agents are of great interest. Sometimes, combination therapy is needed to avoid resistance development in difficult-to-treat infections or biofilm-associated infections treated with common biocides. Thus, this contribution reviews the literature reports discussing the usage of antimicrobial polymers along with nanomaterials or other inhibitors for the development of more potent biocidal therapies.

Kinetic, mechanism and mass transfer impact on electrochemical oxidation of MIT using Ti-enhanced nanotube arrays/SnO2-Sb anode

In this study, a novel Ti-enhanced nanotube arrays/SnO2-Sb electrode was prepared by the sol-gel method on enhanced TiO2 nanotube arrays (ENTA). The characterization, degradation performance, oxidation mechanism and mass transfer impact of the novel electrode was investigated. Compared with the conventional Ti/SnO2-Sb electrode, our novel electrode has higher oxygen evolution potential and electrochemical stability, due to the fabrication of ENTA structure. Its application for the destruction of a common biocide of MIT (2-methyl-4-isothiazolin-3-one), the first-order rate constants were determined from 0.01 min−1 to 0.505 min−1 during the electrochemical process. The radical oxidation and non-radical oxidation of the MIT degradation contribution were quantitatively identified as 87.8% and 12.1%, respectively. Furthermore, the second-rate constant between hydroxyl radical and MIT was estimated at 3.72×109M−1S−1. The mass transfer impact was investigated with a differential column batch reactor (DCBR) under the conditions of various electrode spacing and fluid velocities. It indicated that reducing the electrode spacing and increasing the fluid velocity would enhance the mass transfer process and increase the overall oxidation rate. Meanwhile, the EE/O decreased from 17.7 kWh m−3 to 9.7 kWh m−3, and the mass transfer coefficients increased dramatically from 0.52 × 10−6 m s−1 to 2.48 × 10−6 m s−1.

The use of bacteriophage MS2 for the development and application of a virucide decontamination test method for porous and heavily soiled surfaces.

(i) To develop an analytical method for recovery and quantification of bacteriophage MS2-as a surrogate for foot-and-mouth disease virus-from complex porous surfaces, with and without the presence of laboratory-developed agricultural grime; (ii) to evaluate, with a 4-log dynamic range, the virucidal activity of common biocides for their ability to decontaminate surfaces and hence remediate facilities, following a foreign animal disease contamination incident. An analytical method was developed and optimized for MS2 recovery from simulated agricultural surfaces. The addition of Dey-Engley neutralizing broth to an extraction buffer improved MS2 viability in liquid extracts, with optimal analytical holding times determined as <8 to ≤24h, depending on matrix. The recovery of MS2 from surface materials decreased in the order: nonporous reference material >grimed porous materials >nongrimed porous materials. In disinfectant testing, two spray applications of pAB were effective against MS2 (≥4-log reduction) on all operational-scale materials. Two per cent citric acid had limited effectiveness, with a ≥4-log reduction observed on a selected subset of grimed concrete samples. Decontamination efficacy test results can be affected by surface characteristics, extraction buffer composition, analytical holding time and surface-specific organism survivability. Efficacy should be evaluated using a test method that reflects the environmental characteristics of the intended application. The results of this study demonstrate the importance of analytical method verification tests for disinfectant testing prior to application in complex environments.

Effects of sub-lethal concentrations of copper ammonium acetate, pyrethrins and atrazine on the response of Escherichia coli to antibiotics

Background: Antibiotic resistance in human and animal pathogens is mainly the outcome of human use of antibiotics. However, bacteria are also exposed to thousands of other antimicrobial agents. Increasingly those exposures are being investigated as co-selective agents behind the rapid rise and spread of resistance in bacterial pathogens of people and our domesticated animals. Methods: We measured the sub-lethal effects on antibiotic tolerance of the human pathogen/commensal Escherichia coli caused by exposure to three common biocide formulations based on either copper, pyrethrins, or atrazine as active ingredients. The influence of the efflux pump AcrAB-TolC was investigated using deletion strains, and the persistence of observed effects was determined. Results: Some effects were seen for all biocides, but the largest effects were observed with copper in combination with the antibiotic tetracycline. The effect was caused by both the induction of the adaptive efflux system and by chelation of the antibiotic by copper. Finally, persistence of the adaptive response was measured and found to persist for about two generations. Conclusions: Through a combination of microbe-chemical and chemical-chemical interactions, humanity may be creating micro-environments in which resistance evolution is accelerated.

Effects of sub-lethal concentrations of copper ammonium acetate, pyrethrins and atrazine on the response of Escherichia coli to antibiotics.

Background: Antibiotic resistance in human and animal pathogens is mainly the outcome of human use of antibiotics. However, bacteria are also exposed to thousands of other antimicrobial agents. Increasingly those exposures are being investigated as co-selective agents behind the rapid rise and spread of resistance in bacterial pathogens of people and our domesticated animals. Methods: We measured the sub-lethal effects on antibiotic tolerance of the human pathogen/commensal Escherichia coli caused by exposure to three common biocide formulations based on either copper, pyrethrins, or atrazine as active ingredients. The influence of the efflux pump AcrAB-TolC was investigated using deletion strains, and the persistence of observed effects was determined. Results: Some effects were seen for all biocides, but the largest effects were observed with copper in combination with the antibiotic tetracycline. The effect was caused by both the induction of the adaptive efflux system and by chelation of the antibiotic by copper. Finally, persistence of the adaptive response was measured and found to persist for about two generations. Conclusions: Through a combination of microbe-chemical and chemical-chemical interactions, humanity may be creating micro-environments in which resistance evolution is accelerated.

A sustainable solution for removal of glutaraldehyde in saline water with visible light photocatalysis

Glutaraldehyde (GA) is the most common biocide used in unconventional oil and gas production. Photocatalytic degradation of GA in brine simulating oil and gas produced water using Ag/AgCl/BiOCl composite as a photocatalyst with visible light was investigated. Removal of GA at 0.1 mM in 200 g/L NaCl solution at pH 7 was 90% after 75 min irradiation using 5 g/L of the photocatalyst. The GA removal followed pseudo-first order reaction with a rate constant of 0.0303 min−1. At pH 5 or at 300 g/L NaCl, the photocatalytic removal of GA was almost completely inhibited. Similar inhibitions were observed when adding dissolved organic carbon (from humic acid) at 10 and 200 mg/L, or Br− at 120 mg/L to the system. The removal rate of GA markedly increased with increasing pH (5–9), photocatalyst loading (2–8 g/L) and under 350 nm UV (compared to visible light). On the contrary, the removal rate of GA markedly decreased with increasing NaCl and initial GA concentrations (0–300 g/L for NaCl and 0.1–0.4 mM for GA). A quenching experiment was also conducted; electron holes (h+) and superoxide (▪) were found as the main reactive species responsible for the removal of GA while OH had a very limited effect.

Optimization of microorganism preservation conditions for the development of an acute toxicity bioassay for biocides

Biocides, also referred to as ‘microbicides’ or ‘inhibitors’, are widely used in industrial processes (e.g. utility water in cooling towers) to control and/or eliminate the growth of microorganisms. Because of their inherent toxicity, their presence in various sources (e.g. river sediments, potable water) can negatively affect ecosystems. Currently available biocide detection techniques are not suitable for ‘point-of-use’ applications since they are tedious, complicated, and often require experienced personnel to operate. To address this concern, we sought to develop a simple-to-use toxicity bioassay based on a model microorganism (E. coli) after short (<30 min) exposure to known biocides that can be stored at room temperature (preferably) or in the fridge. Based on recent work and our expertise in polymer-based preservation of biomolecules, we leveraged this knowledge to improve E. coli preservation for biocide detection purposes. A design-of-experiments strategy was used to evaluate 16 different preservation conditions from 5 process parameters (i.e. 25−1 fractional factorial). It was found that pullulan, a sugar-based polymer, improved E. coli culturability by an order of magnitude after three months of storage. Also, it was found that storing E. coli in the fridge in Milli-Q water was favorable for maintaining a high level of culturability. Finally, the toxicity of three common biocides (Cetyltrimethylammonium bromide (CTAB), ProClin™ 300, and Grotan® BK) was evaluated using a fluorescence-based assay across all 16 preservation conditions. The response of the preserved E. coli was biocide specific and at certain conditions did not vary during the entire three-month storage period.