Antifungal and genotoxic effects of Thymus serpyllum as a root canal irrigant.

OBJECTIVES: The aim of this study was the assessment of the efficiency of the ethyl acetate (EthOAc) extract of Thymus serpyllum against Candida albicans and to compare it with sodium hypochlorite (NaOCl) and chlorhexidine (CHX), as well as their genotoxic effect. MATERIAL AND METHODS: The antifungal effectiveness of the EthOAc extract of Thymus serpyllum was determined using the agar disk diffusion method. The inhibition zones induced by the EthOAc extract were compared after 5 min, 60 min, and 24 h to those induced by standard solutions (2% CHX and 2% NaOCl). An in vitro genotoxicity assay was performed in cultured lymphocytes from the blood of human volunteers to observe micronuclei formation. Statistical analysis of the results was performed using the Kruskal-Wallis test and one-way analysis of variance. RESULTS: The inhibition zone of combination of CHX with EthOAc extract of Thymus serpyllum against C. albicans was 29.7 mm after 5 min, 28.3 mm after 60 min, and 29 mm after 24 h. The inhibition zone of NaOCl in combination with EthOAc extract of Thymus serpyllum against C. albicans was 0 mm. The EthOAc extract of Thymus serpyllum did not show a genotoxic effect on lymphocyte cells. CONCLUSIONS: The EthOAc extract of Thymus serpyllum in combination with CHX may be a useful root canal disinfection in endodontic therapy.

Calcium hypochlorite cytotoxicity mechanism in fibroblasts and effect on osteoblast mineralization.

AIM: To determine the cytotoxicity mechanism of 2.5% calcium hypochlorite [Ca(OCl)2 ] in L929 fibroblasts and the effect of this solution on human osteoblast-like cells (Saos-2) mineralization, compared to that of 2.5% sodium hypochlorite (NaOCl). METHODOLOGY: L929 fibroblasts were exposed to Ca(OCl)2 and NaOCl at different dilutions for 10 min. Cell metabolism was assessed by methyl-thiazole-tetrazolium (MTT); lysosome integrity, by neutral red (NR) assay; type of cell death, by flow cytometry (apoptosis/necrosis); cytoskeleton, by actin and α-tubulin fluorescence and cell ultrastructure, by transmission electron microscopy (TEM). The alkaline phosphatase (ALP) activity and mineralized nodule formation were determined in Saos-2 by thymolphthalein release and alizarin red staining (ARS), respectively. The data were analysed by two-way anova and Bonferroni's post-test (α = .05). RESULTS: Ca(OCl)2 promoted higher cell viability and a lower percentage of apoptosis and necrosis than NaOCl (p < .05). Ca(OCl)2 and NaOCl decreased cell metabolism and lysosome integrity, induced the breakdown of microtubules and actin filaments, promoted alterations of rough endoplasmic reticulum and disruption of mitochondrial cristae. Additionally, Ca(OCl)2 did not induce ALP activity and had no effect on mineralized nodules formation. CONCLUSIONS: Although Ca(OCl)2 and NaOCl promoted the same cytotoxicity mechanism, Ca(OCl)2 was less cytotoxic than NaOCl. As for ALP activity, no differences were observed between NaOCl and Ca(OCl)2 . The production of mineralized nodules induced by Ca(OCl)2 was lower than those induced by NaOCl, but was not different from those induced by the control group.

Toxicidad del hipoclorito de sodio y compuestos de amonio cuaternario en seres humanos. Una revisión sistemática en época de COVID-19 / Toxicity of sodium hypochlorite and quaternary ammonium compounds in humans. A systematic review in the time of COVID-19

Los desinfectantes son productos que se utilizan de manera cotidiana, su uso se ha incrementado debido a los cambios en los hábitos de bioseguridad que incorporó la población desde el inicio de la pandemia por la COVID-19; en similar medida, han aumentado las intoxicaciones por la exposición a estas sustancias. Conocer los mecanismos a través de los cuales los productos de desinfección pueden causar una intoxicación es importante para desarrollar estrategias de prevención y manejo oportuno. Con este fin, se revisó la información de artículos científicos y reportes de caso, mediante la búsqueda sistemática en línea en PubMed, Medline, Google Scholar y EBSCO, relacionada a intoxicaciones con compuestos de amonio cuaternario e hipoclorito de sodio. El procedimiento utilizado implicó la búsqueda de palabras claves alusivas al tema mediante el uso de Descriptores en Ciencias en Salud y Medical Subject Headings, posteriormente se utilizó metodología “Preferred Reporting Items for Systematic reviews and Meta-Analyses”. Los resultados del estudio evidenciaron que el hipoclorito de sodio afecta a las células y tejidos, mediante sus propiedades oxidativas, provocando alteración de la membrana citoplasmática, desnaturalización y formación de agregados en las proteínas, daño al ADN e inflamación asociada a la lisis celular. Por otro lado, los compuestos de amonio cuaternario afectan principalmente a la membrana celular, ocasionando daño inflamatorio, fibrosis y alteraciones reproductivas. Los dos compuestos afectan la integridad de la célula de manera directa. Sin embargo, sus efectos se ven exacerbados por los cambios que ocurren en el entorno celular. (AU)

Disinfectants are products that are used on a daily basis, their use has increased due to changes in biosafety habits that the population has adopted since the beginning of COVID19. To a similar extent, poisoning from exposure to these substances has increased. Consequently, knowing the mechanisms through which disinfection products can cause poisoning is important to develop prevention strategies and timely management. To this end, the information related to poisoning with quaternary ammonium compounds and sodium hypochlorite was reviewed in scientific articles and study cases by a systematic online search in PubMed, Medline, Google Scholar and EBSCO. The procedure used involved the search for keywords alluding to the topic by using Descriptors in Health Sciences and Medical Subject Headings. Later the «Preferred Reporting Items for Systematic reviews and Meta-Analyses» methodology was used. The results of the study showed that sodium hypochlorite affects cells and tissues through its oxidative properties, causing alteration of the cytoplasmic membrane, denaturation and formation of protein aggregates, DNA damage and inflammation associated with cell lysis. On the other hand, quaternary ammonium compounds mainly affect the cell membrane, causing inflammatory damage, fibrosis and reproductive disorders. Both compounds directly affect the integrity of the cell. However, its effects are exacerbated by changes that occur in the cellular environment. (AU)

Assessment of acute inhalation toxicity of citric acid and sodium hypochlorite in rats.

BACKGROUND: Citric acid (CA) and sodium hypochlorite (NaOCl) have been used to disinfect animals to protect them against avian influenza and foot-and-mouth disease. OBJECTIVES: We performed a good laboratory practice (GLP)-compliant animal toxicity study to assess the acute toxic effects of CA and NaOCl aerosol exposure in Sprague-Dawley rats. METHODS: Groups of five rats per sex were exposed for 4 h to four concentrations of the two chemicals, i.e., 0.00, 0.22, 0.67, and 2.00 mg/L, using a nose-only exposure. After a single exposure to the chemicals, clinical signs, body weight, and mortality was observed during the observation period. On day 15, an autopsy, and then gross findings, and histopathological analysis were performed. RESULTS: After exposure to CA and NaOCl, body weight loss was observed but recovered. Two males died in the CA 2.00 mg/L group and, two males and one female died in the 2.00 mg/L NaOCl group. In the gross findings and histopathological analysis, discoloration of the lungs was observed in the CA exposed group and inflammatory lesions with discoloration of the lungs were observed in the NaOCl exposed group. These results suggest that the lethal concentration 50 (LC50) of CA is 1.73390 mg/L for males and > 1.70 mg/L for females. For NaOCl, the LC50 was 2.22222 mg/L for males and 2.39456 mg/L for females. CONCLUSIONS: The Globally Harmonized System is category 4 for both CA and NaOCl. In this study, the LC50 results were obtained through a GLP-based acute inhalation toxicity assessment. These results provide useful data to reset safety standards for CA and NaOCl use.

Acute inhalation toxicity of aerosolized electrochemically generated solution of sodium hypochlorite.

OBJECTIVE: The objective was to determine the inhalation toxicity of the electrochemically generated sodium hypochlorite solution after its single administration to laboratory animals in the form of a highly dispersed aerosol. MATERIALS AND METHODS: The study has been conducted according to the OECD Test Guideline №403 'Acute Inhalation Toxicity.' Laboratory animals were exposed to inhalation of an aerosol containing 1.7 ± 0.13 mg/m3 of active chlorine. The hematological and biochemical parameters of the blood of experimental animals have been determined, as well as specific parameters: the activity of cathepsins B and L, catalase, and α1-antitrypsin. Histological study of the lungs of animals has been carried out. RESULTS: During inhalation and 14 days after it, no death of the animals was observed; the behavior, appearance, and weight gain did not differ from the control group. There were no significant deviations in hematological parameters, except the decrease in the level of platelets. The biochemical study showed slight changes in the activity of alkaline phosphatase and aspartate aminotransferase on the 1st day after inhalation; these parameters returned to normal within 14 days of observation. Specific biochemical parameters did not show the development of oxidative stress. No specific histological pathologies of lung tissue have been found. CONCLUSIONS: Thus, the studied electrochemically generated sodium hypochlorite solution under single inhalation exposure in aerosol form practically does not cause a toxic effect. The data obtained allow classifying such solution to the 4th (or even 5th - after additional studies) class of toxicity in accordance with Globally Harmonized System of Classification and Labeling of Chemicals.

Cytotoxic effect of sodium hypochlorite (Lavanox 0.08%) and chlorhexidine gluconate (Irrisept 0.05%) on human osteoblasts.

PURPOSE: Soft tissue, bone and joint infections are severe complications in orthopedic and traumatological surgery. Lavanox (0.08% NaOCl) and Irrisept (0.05% chlorhexidine gluconate, CHG) are industrially produced antiseptic solutions commonly used in infection treatment. Regarding this clinical indication, the microbicidal effect is often investigated, but toxicity to osteoblasts has rarely been examined. This is important to decide whether these solutions should be used in septic situations in which bone healing must take place. The hypothesis of the present study is that NaOCl and CHG are cytotoxic to osteoblasts even after a short exposure time. METHODS: Human osteoblasts were isolated from donors with osteoarthritis during total knee and hip arthroplasty. Cells were cultivated and treated with both antiseptic solutions for 2, 5 and 10 min in different dilutions. Toxicity was quantified by counting cells, lactate dehydrogenase (LDH) expression, spectrophotometric quantification via XTT assay and FDA/PI fluorescence microscopy. RESULTS: Analyzing viable cells after treatment with both antiseptics showed a significant decrease in viable cells through LDH expression test, XTT assay, fluorescence microscopy and light microscopy, depending on concentration. The time dependence showed a trend to more cell death at longer exposure times, without significance. CONCLUSION: Toxic effects on osteoblasts were shown after treatment with 0.08% NaOCl and 0.05% CHG after an exposure time of 2 min which also was concentration dependent. There was no difference in cytotoxicity between both antiseptics. In conclusion, these antiseptic solutions may be used with caution in situations requiring bone healing. Trial registration number Local ethics committee registration number: 5176-07/16.

The cytotoxic effects of various endodontic irrigants on the viability of dental mesenchymal stem cells.

This study aimed to evaluate cytotoxic effects of various irrigation solutions used in regenerative endodontic treatments (RETs) on mesenchymal stem cells, and further examine the long-term effect of hypochlorous acid (HOCl) on the cell viability and alkaline phosphatase (ALP) activity. Stem cells were exposed to various concentrations of NaOCl, EDTA, chlorhexidine (CHX), etidronic acid (HEDP)/NaOCl combination and HOCl. HOCl was tested for its effects on ALP activity up to 21 days. Additionally, cell viability was measured fluorescently using calcein AM. The most cytotoxic irrigant was CHX even with the lowest concentration. NaOCl and HEDP/NaOCl with 1:100 dilution decreased viability to around 40%. HOCl showed the lowest cytotoxicity among all tested irrigants. HOCl also showed no significant reduction in ALP activity compared with the controls. The cytotoxicity of endodontic irrigants was time and concentration dependent. HOCl demonstrated promising results regarding viability and ALP activity, since RETs require host stem cell survival.

Assessment of the cytotoxic effects and chemical composition of the insoluble precipitate formed from sodium hypochlorite and chlorhexidine gluconate.

AIM: To investigate (1) the cytotoxic potential of the brown precipitate (BP) formed with sodium hypochlorite (NaOCl) and chlorhexidine gluconate (CHX), using both a small animal model of Caenorhabditis elegans (C. elegans) and cultured human gingival fibroblasts; and (2) the chemical composition of BP using Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS). METHODOLOGY: Brown precipitate was obtained by mixing equal volumes of 6% NaOCl and 2% CHX and separating the BP from clear supernatant by centrifugation. The brown precipitate was weighed and solubilized in dimethyl sulfoxide for cytotoxicity experiments. The cytotoxic effect of BP was assessed using C. elegans larvae and primary immortalized human gingival fibroblasts-hTERT (hTERT-hNOF) cells. Various dilutions of BP (25 ng/µL-150 ng/µL), supernatant (0.15% v/v), NaOCl (1:100-1:1000 dilutions of 6% NaOCl) or CHX (1:500-1:1000 dilutions of 2% CHX) along with vehicle control (0.5% v/v ethanol and 0.15% v/v DMSO) or untreated control (growth medium) were tested on C. elegans larvae and hTERT-hNOF cells. Viability was assessed in C. elegans larvae using stereomicroscopy and in hTERT-hNOF cells using dehydrogenase-based colorimetric assay. ToF-SIMS was used to assess the chemical composition of BP in comparison with CHX and para-chloroaniline (PCA). The C. elegans and cell line data were analysed using Log-Rank test and Student's t-test, respectively (p < .05). RESULTS: BP-75 ng/µL and BP-150 ng/µL were significantly more toxic to C. elegans larvae than the untreated, vehicle, supernatant or CHX treatment groups (p < .0001). Similarly, in hTERT-hNOF cells, BP-50 ng/µL, BP-75 ng/µL and BP-150 ng/µL induced significant cytotoxicity within 2 h compared with untreated, vehicle, supernatant and CHX treatments (p < .05). ToF-SIMS analysis of BP revealed ion composition characteristic of both CHX and the carcinogen PCA. CONCLUSIONS: Brown precipitate was toxic in both C. elegans larvae and hTERT-hNOF cells. The ToF-SIMS analysis of BP revealed ions characteristic of CHX and PCA that could account for the toxicities observed in C. elegans larvae and human gingival fibroblasts. Because of the insoluble and toxic nature of BP, consecutive use of CHX and NaOCl irrigants should be avoided in root canal treatment.

Cytotoxicity evaluation of calcium hypochlorite and other commonly used root canal irrigants against human gingival fibroblast cells: An in vitro evaluation.

BACKGROUND: The conventional endodontic therapy primarily focuses on biomechanical preparation, which is achieved by the application of various intracanal irrigants and intracanal medicaments. One of the most commonly used intracanal irrigants - sodium hypochlorite (NaOCl) - has already been proven to have an antimicrobial effect as well as the ability to dissolve tissues in the areas where files cannot reach. One of the recently used irrigants having a promising effect is calcium hypochlorite (Ca(OCl)2), which has been shown to be relatively more stable than NaOCl and has much more chlorine ions. OBJECTIVES: The aim of this study was to assess the individual cytotoxicity of various root canal irrigants and the combined cytotoxicity of NaOCl and Ca(OCl)2 with ethylenediaminetetraacetic acid (EDTA) against human gingival fibroblast (hGF) cells. MATERIAL AND METHODS: The evaluation of the individual cytotoxicity was carried out with regard to the following root canal irrigants: NaOCl; Ca(OCl)2; and chlorhexidine (CHX). The evaluation of the combined cytotoxicity regarded NaOCl/EDTA and Ca(OCl)2/EDTA. The concentrations used were 0.025%, 0.050%, 0.10%, and 0.20%. The cytotoxicity against hGF cells was examined within a timeframe of 6 h and 24 h with the use of the sulforhodamine B (SRB) assay. RESULTS: It was observed that Ca(OCl)2 had a mean absorbance rate of 0.315 ±0.02, 0.294 ±0.03, 0.265 ±0.03, and 0.240 ±0.02 at 0.025%, 0.050%, 0.10%, and 0.20%, respectively. In combination with EDTA, the mean absorbance rate was 70.12 ±2.9, 67.42 ±4.3, 64.35 ±3.6, and 61.58 ±4.1 at 0.025%, 0.050%, 0.10%, and 0.20%, respectively. The cytotoxic effect of the root canal irrigants on hGF cells was observed to be statistically significant (p < 0.05). CONCLUSIONS: Calcium hypochlorite is less cytotoxic than NaOCl, and when used in combination with EDTA, it was shown to have its cytotoxic effect on hGF cells reduced to a great extent.

Mitigation of graphene oxide toxicity in C. elegans after chemical degradation with sodium hypochlorite.

Graphene oxide (GO) is a promising and strategic carbon-based nanomaterial for innovative and disruptive technologies. It is therefore essential to address its environmental health and safety aspects. In this work, we evaluated the chemical degradation of graphene oxide by sodium hypochlorite (NaClO, bleach water) and its consequences over toxicity, on the nematode Caenorhabditis elegans. The morphological, chemical, and structural properties of GO and its degraded product, termed NaClO-GO, were characterized, exploring an integrated approach. After the chemical degradation of GO at room temperature, its flake size was reduced from 156 to 29 nm, while NaClO-GO showed changes in UV-vis absorption, and an increase in the amount of oxygenated surface groups, which dramatically improved its colloidal stability in moderately hard reconstituted water (EPA medium). Acute and chronic exposure endpoints (survival, growth, fertility, and reproduction) were monitored to evaluate material toxicities. NaClO-GO presented lower toxicity at all endpoints. For example, an increase of over 100% in nematode survival was verified for the degraded material when compared to GO at 10 mg L-1. Additionally, enhanced dark-field hyperspectral microscopy confirmed the oral uptake of both materials by C. elegans. Finally, this work represents a new contribution toward a better understanding of the links between the transformation of graphene-based materials and nanotoxicity effects (mitigation), which is mandatory for the safety improvements that are required to maximize nanotechnological benefits to society.

Oxidation of diclofenac in water by sodium hypochlorite: Identification of new degradation by-products and their ecotoxicological evaluation.

Diclofenac (DCF) is the most widely prescribed non-steroidal anti-inflammatory drug in the world and it has been detected in drinking and surface waters. In this paper, the effect of chlorination process on DCF in aqueous solutions was investigated and the structures of 14 isolated degradation by-products (DPs), of which nine are new, have been determined from combining mass spectrometry and nuclear magnetic resonance data and justified by a proposed mechanism of formation beginning from the parent drug. Some degradation by-products show only one phenyl, others are dimers or trimers of the parental compound, which has undergone oxidative decarboxylation of the side chain and/or chlorination of this or one or both aromatic rings. Ecotoxicological bioassays evidenced the following sensitivities D. magna < R. subcapitata < A. fischeri. The isolated DPs (DP1-8, except for DP9) exhibited effects ≥ 50 % in the exposed microalgae and crustaceans showing toxicities mainly ranked from slight to acute.

¿Puede la ingesta de lavandina o lejía prevenir o curar la COVID-19? [infografía]

El uso de Lavandina o lejía (hipoclorito de sodio) sirve para matar gérmenes en superficies inanimadas. No es un producto para ser ingerido o usado en personas.

Calcium hypochlorite on mouse embryonic fibroblast cells (NIH3T3) in vitro cytotoxicity and genotoxicity: MTT and comet assay.

Antimicrobial irrigation solutions are widely used under clinical settings. Their effect on dental tissue is a subject of recent research, which aims for a safer irrigant for clinical use. In this regard, here our goal was to evaluate the cytotoxicity and the genotoxicity of calcium hypochlorite (Ca(OCl)2) solution, along with NaOCl, on Mouse embryonic fibroblast cells (NIH3T3). First, Cells were treated either with NaOCl or Ca(OCl)2 in a time- and dose-dependent manner for cytotoxicity by 3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, then cell viability was calculated according to cell proliferation plots. Secondly, genotoxicity was assessed by Comet assay. Data were statistically analyzed by Tukey's test (P < .05). NaOCl and Ca(OCl)2 had similar effects on cellular viability at 3 and 6 h treatments. Cell viability of Ca(OCl)2 at concentrations of 0.0125%, 0.025%, 0.05%, or 0.125% was significantly lower than that of NaOCl at 24 h treatment (P < .05).Comparing Ca(OCl)2 and NaOCl treatments at all time points and concentrations, the damaged cell number of Ca(OCl)2 was almost fourfold higher than that of NaOCl. In conclusion, both, NaOCl and Ca(OCl)2 solutions were cytotoxic and genotoxic to NIH3T3, however, Ca(OCl)2 had a significantly higher damaged cell percentage than NaOCl at all time points and concentrations investigated.

Evaluation of the cytotoxic potential of sodium hypochlorite using meristematic root cells of Lens culinaris Med.

The present investigation was designed to monitor the cytotoxic potential of Sodium Hypochlorite using lentil (Lens culinaris) as a bioindicator of toxicity. Sodium hypochlorite (NaClO), is a chemical compound that is used mainly for its disinfectant properties, its effect is widely toxic, which is why it is marketed in low concentrations and it is also a component in various products such as agrochemicals. In the present study the L. culinaris seeds were exposed to different NaClO dose 0, 0.2, 1, 3, 5 and 7 mg L-1 during 24, 48 and 72 h; timeslots in which the root growth was also studied. The cytotoxic potential of NaClO was determined by calculating the mitotic index (MI), calculating cellular anomalies (CA) and observing the longitudinal growth of the roots during the various time periods. The radicular growth was prolonged and it was observed that there was a greater growth at the dose of 1 and 7 mg L-1 in the time of 72 h. The cytotoxic effects could be analyzed in the mitotic index, since the higher the concentration, the lower the mitotic index, as observed in the dose of 7 mg L-1 where a reduction of the mitotic index of the meristematic cells is observed. The results indicate that NaClO has a cytotoxic effect that induces various types of chromosomal abnormalities. This indicates that Sodium Hypochlorite has a cytotoxic effect according to the increase in its dose. Therefore, Lens culinaris turned out to be a kind of appropriate bioindicator to study the cytotoxic effects of various potentially toxic substances.

Comparison of cytotoxicity and smear layer removal efficacy of triphala (an Indian ayurvedic herbal formulation) and 5.25% sodium hypochlorite as root canal irrigants: An in vitro study.

INTRODUCTIO: Healing potential of plants is an age-old idea that has recently attained renewed interest. Considering the ineffectiveness, potentially harmful effects, and safety concerns of commonly used synthetic irrigants, the herbal alternatives for endodontic usage might prove to be advantageous. AIM: The aim of this study was to evaluate the adequacy of smear layer removal and cytotoxicity potential of triphala in comparison to sodium hypochlorite. MATERIALS AND METHODS: The study was conducted in two parts: the first part of the study was cytotoxicity assessment studied using Alamar blue assay. L929 mouse fibroblasts were seeded in 96-well plates at a density of 5000 cells/well and treated with different concentrations of triphala and NaOCl for a period of 24 and 48 h. The percentage of cell viability was then quantified using an Alamar blue assay. The optical density was measured at 570 nm and compared with 620 nm, which was considered as a reference wavelength. The second part of the study was smear layer assessment at the coronal, middle, and apical third of twenty human premolar teeth using scanning electron microscope. RESULTS: The Alamar blue reagent cytotoxicity study suggested that triphala showed no cytotoxic properties against the normal mouse fibroblast cells whereas sodium hypochlorite showed a significant cytotoxic effect against the L929 cell lines with the IC50 concentration at 1.8%, respectively, after the treatment of 24 h of incubation at 37°C temperature. Triphala was as effective as sodium hypochlorite in smear layer removal in the coronal and middle third of the root, but sodium hypochlorite showed better smear layer removal in the apical third. CONCLUSION: Triphala can be considered as a superior irrigant with good antibacterial efficacy and least cytotoxicity potential compared to conventional hypochlorite irrigating agent and provide adequate clearing of smear layer in the coronal and middle third, and further studies are warranted to alter the properties of liquid to make it more cleansable in the apical third of the root.

Development, standardization, and validation of a biofilm efficacy test: The single tube method.

Methods validated by a standard setting organization enable public, industry and regulatory stakeholders to make decisions on the acceptability of products, devices and processes. This is because standard methods are demonstrably reproducible when performed in different laboratories by different researchers, responsive to different products, and rugged when small (usually inadvertent) variations from the standard procedure occur. The Single Tube Method (ASTM E2871) is a standard method that measures the efficacy of antimicrobials against biofilm bacteria that has been shown to be reproducible, responsive and rugged. In support of the reproducibility assessment, a six-laboratory study was performed using three antimicrobials: a sodium hypochlorite, a phenolic and a quaternary/alcohol blend, each tested at low and high efficacy levels. The mean log reduction in viable bacteria in this study ranged from 2.32 to 4.58 and the associated reproducibility standard deviations ranged from 0.89 to 1.67. Independent follow-up testing showed that the method was rugged with respect to deviations in sonication duration and sonication power but slightly sensitive to sonicator reservoir degassing and tube location within the sonicator bath. It was also demonstrated that when a coupon was dropped into a test tube, bacteria can splash out of reach of the applied antimicrobials, resulting in substantial bias when estimating log reductions for the products tested. Bias can also result when testing products that hinder the harvesting of microbes from test surfaces. The culmination of this work provided recommended changes to the early version of the standard method E2871-13 (ASTM, 2013b) including use of splashguards and microscopy checks. These changes have been incorporated into a revised ASTM method E2871-19 (ASTM 2019) that is the basis for the first regulatory method (ATMP-MB-20) to substantiate "kills biofilm" claims for antimicrobials registered and sold in the US.

Comparative study on the efficacy of sodium hypochlorite, aqueous ozone, and peracetic acid in the elimination of Salmonella from cattle manure contaminated various surfaces supported by Bayesian analysis.

Providing the dairy industry with an effective and safe disinfectant is considered a key step in improving the farm hygiene and biosecurity. Salmonella infection via foodborne transmission remains a major public health threat. The main objective of this study was therefore to characterize and compare the decontamination power of NaOCl, aqueous-O3, and PAA against cattle manure based-Salmonella heavily contaminated various surfaces (plastic, nylon, rubber, and wood) using Bayesian analysis. In a crossover design, 14 strips of each material were randomly assigned between 3 groups, treatment (n = 6), positive-control (contaminated with feces-Salmonella mixture, but not exposed to disinfectants, n = 6), and negative control (laboratory blank, inoculated only with sterile water, n = 2). The strips were soaked in cattle manure inoculated with 107-108 of Salmonella Typhimurium-Choleraesuis (aSTC) and exposed to 50 mL of 200 ppm NaOCl, 9 ppm aqueous-O3, or 400 ppm PAA for 4 minutes. Bayesian methods were used for analysis. On plastic and nylon surfaces, NaOCl, aqueous-O3, or PAA reduce aSTC population to a safe level (>5.0-log10) within 4 minutes. On rubber surface, PAA and aqueous-O3 can produce a reduction in aSTC population 50% and 30% higher than NaOCl with posterior probabilities of 97% and 90%, respectively. However, PAA can produce reduction factor on wood surface 40% higher than aqueous-O3 and NaOCl with posterior probabilities of 97% and 73%, respectively. We conclude that smooth surfaces were most effectively decontaminated. Peracetic acid of 400 ppm can provide an effective means for controlling Salmonella population heavily contaminated various surfaces in dairy operations. However, the safe residues and strong reactivity makes aqueous-O3 and PAA attractive alternative disinfectants for improving farm hygiene and biosecurity.

Effects of sodium hypochlorite concentration on the methanogenic activity in an anaerobic fluidized membrane bioreactor.

The effect of membrane cleaning chemical, NaOCl on specific acetoclastic methanogenic activity (SAMA) of biomass in the anaerobic fluidized membrane bioreactors (AFMBRs) was assessed. Granular activated carbon (GAC) was used as a fluidizing media in the AFMBR to provide membrane scouring effect and surface for biofilm attachment. Effect of NaOCl on methane production was negligible for GAC with biofilm (bGAC) samples up to 150 mg NaOCl/g VSS, but was significant for the bulk liquid samples with noticeable lag period over 300 h even at the dosage of 50 mg NaOCl/g VSS. The toxicity of NaOCl on methane production was reduced in samples with virgin GAC (vGAC) by allowing 14 days of biomass buildup period prior to NaOCl addition, although the vGAC alone did not alleviate the toxicity. The results revealed that NaOCl concentrations beyond 100 mg/L within the reactor inhibited methanogenic activity and the effects were more pronounced on suspended biomass than the immobilized biomass on GAC.

Cytotoxic evaluation of sodium hypochlorite, using Pisum sativum L as effective bioindicator.

The objective of this study was to evaluate the cytotoxic effect of different sodium hypochlorite concentrations, using apical root cells of P. sativum as a bioindicator. Initially, the seeds of P. sativum were exposed to different concentrations of sodium hypochlorite (0.1, 0.2, 0.4, 0.8, 1.6, 2 mg L-1) and to a control solution based on distilled water. Next, root growth was measured during 24, 48 and 72 h. Subsequently, the mitotic index (MI) and cellular anomalies (5000 cells per treatment) were determined at 72 h. According to the results obtained, a decrease in root growth was observed at concentrations of 0.4, 1.6 and 2 mg L-1. Likewise, it was evident that, among all the evaluated concentrations, an inhibition of mitosis higher than 50% was presented. Additionally, chromosomal anomalies were also generated, such as Nuclear notch, lagging chromosomes and Chromosomal break, which were present in all the concentrations evaluated. In addition, the presence of micronuclei at concentrations of 2.0 and 1.6 mg L-1 indicate that sodium hypochlorite is a highly cytotoxic substance. Therefore, P. sativum is a specie that offers a feasible experimental model to be implemented in the laboratory with the aim to evaluate the cytotoxic effect of any cytotoxic substance.

Assessment of toxicity and oxidative DNA damage of sodium hypochlorite, chitosan and propolis on fibroblast cells.

The objective of this study was to evaluate and compare the cytotoxicity and genotoxicity on human fibroblast cell lines of sodium hypochlorite (NaOCl), chitosan and propolis as root canal irrigating solutions. Human fibroblast cells were exposed to chitosan, propolis and NaOCl for 4 and 24 h. Cell viability was assessed by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide, and oxidative DNA damage was assessed by determination of 8-hydroxydeoxyguanosine (8-OHdG) level with an ELISA kit. The data of cell cytotoxicity were analysed statistically using a test of one-way analysis of variance at a significance level of p < 0.05. In the NaOCI group, the 8-OHdG level was higher than in the chitosan group, but there was no statistical difference when compared with the other groups (p < 0.05). It was determined that the irrigation solutions were cytotoxic, depending on the dose and time. NaOCl was the most toxic solution after both 4 and 24 h of exposure (p < 0.05). Chitosan and propolis may be alternatives to NaOCl for irrigation solutions, because they are both less toxic and produce less oxidative DNA damage.