Mutant Prevention Concentration, Frequency of Spontaneous Mutant Selection, and Mutant Selection Window-a New Approach to the In Vitro Determination of the Antimicrobial Potency of Compounds.

The analysis of antimicrobial activity is usually MIC- and minimal bactericidal concentration (MBC)-focused, though also crucial are resistance-related parameters, e.g., the frequency of spontaneous mutant selection (FSMS), the mutant prevention concentration (MPC), and the mutant selection window (MSW). In vitro-determined MPCs, however, are sometimes variable, poorly repeatable, and not always reproducible in vivo. We propose a new approach to the in vitro determination of MSWs, along with novel parameters: MPC-D, MSW-D (for dominant mutants, i.e., selected with a high frequency, without a fitness loss), and MPC-F, MSW-F (for inferior mutants, i.e., with an impaired fitness). We also propose a new method for preparing the high-density inoculum (>1011 CFU/mL). In this study, the MPC and MPC-D (limited by FSMS of <10-10) of ciprofloxacin, linezolid, and novel benzosiloxaborole (No37) were determined for Staphylococcus aureus ATCC 29213 using the standard agar method, while the MPC-D and MPC-F were determined by the novel broth method. Regardless of the method, MSWs1010 of linezolid and No37 were the same. However, MSWs1010 of ciprofloxacin in the broth method was narrower than in the agar method. In the broth method, the 24-h incubation of ~1010 CFU in a drug-containing broth differentiates the mutants that can dominate the cell population from those that can only be selected under exposure. We consider MPC-Ds in the agar method to be less variable and more repeatable than MPCs. Meanwhile, the broth method may decrease discrepancies between in vitro and in vivo MSWs. The proposed approaches may help establish MPC-D-related resistance-restricting therapies.

Efflux-Related Carbapenem Resistance in Acinetobacter baumannii Is Associated with Two-Component Regulatory Efflux Systems' Alteration and Insertion of ΔAbaR25-Type Island Fragment.

The efflux pumps, beside the class D carbapenem-hydrolysing enzymes (CHLDs), are being increasingly investigated as a mechanism of carbapenem resistance in Acinetobacter baumannii. This study investigates the contribution of efflux mechanism to carbapenem resistance in 61 acquired blaCHDL-genes-carrying A. baumannii clinical strains isolated in Warsaw, Poland. Studies were conducted using phenotypic (susceptibility testing to carbapenems ± efflux pump inhibitors (EPIs)) and molecular (determining expression levels of efflux operon with regulatory-gene and whole genome sequencing (WGS)) methods. EPIs reduced carbapenem resistance of 14/61 isolates. Upregulation (5-67-fold) of adeB was observed together with mutations in the sequences of AdeRS local and of BaeS global regulators in all 15 selected isolates. Long-read WGS of isolate no. AB96 revealed the presence of AbaR25 resistance island and its two disrupted elements: the first contained a duplicate ISAba1-blaOXA-23, and the second was located between adeR and adeA in the efflux operon. This insert was flanked by two copies of ISAba1, and one of them provides a strong promoter for adeABC, elevating the adeB expression levels. Our study for the first time reports the involvement of the insertion of the ΔAbaR25-type resistance island fragment with ISAba1 element upstream the efflux operon in the carbapenem resistance of A. baumannii.

Adhesion and aggregation properties of Lactobacillaceae strains as protection ways against enteropathogenic bacteria.

The adhesion and aggregation are characteristic attributes of probiotic strains belonging to Lactobacillaceae genus. Due to these properties the host organisms can avoid colonisation of the intestinal tract by enteropathogenic bacteria. The presented research includes a comparison of the properties of various strains belonging to different Lactobacillaceae species and isolated from different sources The aim of this study was to investigate the ability of Lactocaseibacillus rhamnosus, Lactiplantibacillus plantarum, and Lactobacillus strains (L. acidophilus, L. gasseri, L. ultunensis) from probiotic products and clinical specimens to direct and competitive adherence to Caco-2 and HT-29 cell lines. Furthermore, the ability of lactobacilli and enteropathogenic bacteria, E. coli, E. faecalis, and S. Typhimurium, to auto- and co-aggregation was also investigated.The results showed that all tested strains adhered to Caco-2 and HT-29 cell lines. Though, the factor of adhesion depended on the species and origin of the strain. L. rhamnosus strains showed a lowest degree of adherence as compared to L. plantarum and Lactobacillus sp. strains. On the other side both, L. rhamnosus and L. acidophilus strains reduced the pathogenic bacteria in competition adherence test most effectively. All tested lactobacilli strains were characterised by auto- and co-aggregation abilities, to various degrees. The properties of Lactobacillaceae strains analysed in this study, like adhesion abilities, competitive adherence, auto- and co-aggregation, may affect the prevention of colonisation and elimination of pathogenic bacteria in gastrointestinal tract.

Analysis of blaCHDL Genes and Insertion Sequences Related to Carbapenem Resistance in Acinetobacter baumannii Clinical Strains Isolated in Warsaw, Poland.

Acinetobacter baumannii is an important cause of nosocomial infections worldwide. The elucidation of the carbapenem resistance mechanisms of hospital strains is necessary for the effective treatment and prevention of resistance gene transmission. The main mechanism of carbapenem resistance in A. baumannii is carbapenemases, whose expressions are affected by the presence of insertion sequences (ISs) upstream of blaCHDL genes. In this study, 61 imipenem-nonsusceptible A. baumannii isolates were characterized using phenotypic (drug-susceptibility profile using CarbaAcineto NP) and molecular methods. Pulsed field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) methods were utilized for the genotyping. The majority of isolates (59/61) carried one of the following acquired blaCHDL genes: blaOXA-24-like (39/59), ISAba1-blaOXA-23-like (14/59) or ISAba3-blaOXA-58-like (6/59). Whole genome sequence analysis of 15 selected isolates identified the following intrinsic blaOXA-66 (OXA-51-like; n = 15) and acquired class D ß-lactamases (CHDLs): ISAba1-blaOXA-23 (OXA-23-like; n = 7), ISAba3-blaOXA-58-ISAba3 (OXA-58-like; n = 2) and blaOXA-72 (OXA-24-like; n = 6). The isolates were classified into 21 pulsotypes using PFGE, and the representative 15 isolates were found to belong to sequence type ST2 of the Pasteur MLST scheme from the global IC2 clone. The Oxford MLST scheme revealed the diversity among these studied isolates, and identified five sequence types (ST195, ST208, ST208/ST1806, ST348 and ST425). CHDL-type carbapenemases and insertion elements upstream of the blaCHDL genes were found to be widespread among Polish A. baumannii clinical isolates, and this contributed to their carbapenem resistance.

Are Probiotic Really Safe for Humans?

Probiotic bacteria have been used as a health-promoting factor for a very long time. Nowadays, products containing probiotic bacteria are becoming more and more popular on the market. The term probiotics refers to the products belonging to the following groups: probiotic drugs (medicinal products - live biotherapeutic products for human use), medical devices, probiotic foods (e.g. foods, food ingredients, dietary supplements or food for special medical purposes), directly fed microorganisms (for animal use) and designer probiotics (genetically modified probiotics). Safety assessment of bacterial strains used as probiotics should be carefully studied. Even though probiotic bacteria have the generally recognized as safe (GRAS status), there are several reports about side effects triggered by the presence of these organisms. Microorganisms used as probiotics may cause systemic infections, stimulate the immune system, disturb metabolism and participate in horizontal gene transfer.

The Impact of Efflux Pump Inhibitors on the Activity of Selected Non-Antibiotic Medicinal Products against Gram-Negative Bacteria.

The potential role of non-antibiotic medicinal products in the treatment of multidrug-resistant Gram-negative bacteria has recently been investigated. It is highly likely that the presence of efflux pumps may be one of the reasons for the weak activity of non-antibiotics, as in the case of some non-steroidal anti-inflammatory drugs (NSAIDs), against Gram-negative rods. The activity of eight drugs of potential non-antibiotic activity, active substance standards, and relevant medicinal products were analysed with and without of efflux pump inhibitors against 180 strains of five Gram-negative rod species by minimum inhibitory concentration (MIC) value determination in the presence of 1 mM MgSO4. Furthermore, the influence of non-antibiotics on the susceptibility of clinical strains to quinolones with or without PAßN (Phe-Arg-ß-naphthylamide) was investigated. The impacts of PAßN on the susceptibility of bacteria to non-antibiotics suggests that amitriptyline, alendronate, nicergoline, and ticlopidine are substrates of efflux pumps in Gram-negative rods. Amitriptyline/Amitriptylinum showed the highest direct antibacterial activity, with MICs ranging 100-800 mg/L against all studied species. Significant decreases in the MIC values of other active substances (acyclovir, atorvastatin, and famotidine) tested with pump inhibitors were not observed. The investigated non-antibiotic medicinal products did not alter the MICs of quinolones in the absence and in the presence of PAßN to the studied clinical strains of five groups of species.

Looking for the new preparations for antibacterial therapy. V. New antimicrobial agents from the oxazolidinones groups in clinical trials

This paper is the fifth part of the series concerning the search for new preparations for antibacterial therapy and discussing new compounds belonging to the oxazolidinone class of antibacterial chemotherapeutics. This article presents five new substances that are currently at the stage of clinical trials (radezolid, sutezolid, posizolid, LCB01-0371 and MRX-I). The intensive search for new antibiotics and antibacterial chemotherapeutics with effective antibacterial activity is aimed at overcoming the existing resistance mechanisms in order to effectively fight against multidrug-resistant bacteria, which pose a real threat to public health. The crisis of antibiotic resistance can be overcome by the proper use of these drugs, based on bacteriological and pharmacological knowledge. Oxazolidinones, with their unique mechanism of action and favorable pharmacokinetic and pharmacodynamic parameters, represent an alternative way to effectively treat serious infections caused by Gram-positive microorganisms.

Assessment of the Microbiological Status of Probiotic Products.

The aim of this study was to perform the microbiological analysis of quality of 25 probiotic products, available on the Polish market. Analysis of bacterial viability in probiotic products showed that not all of these preparations possess a suitable number of bacteria. Moreover, some of the tested probiotic products contained bacterial strains other than those declared by the manufacturer. All tested strains recovered from probiotic products were found to be resistant to metronidazole and susceptible to nitrofurantoin. The susceptibility to other antibiotics was strain specific. Probiotic products should be subject to regular and thorough inspection by appropriate institutions.

Activity of Natural Polyether Ionophores: Monensin and Salinomycin against Clinical Staphylococcus epidermidis Strains.

Staphylococcus epidermidis, a coagulase-negative Staphylococcus, is the most important pathogen responsible for chronic nosocomial infections. These bacteria produce extracellular slime and form biofilms on various biotic and abiotic surfaces. Bacterial biofilms are very resistant to standard antimicrobial therapy and difficult to eradicate, so it is important to search for new more effective anti-biofilm agents, for example in the group of natural substances. The aim of the study was to examine the activity of two ionophores-salinomycin and monensin against clinical S. epidermidis strains, using MIC/MBC method and biofilm formation inhibition assay. Bacterial strains were tested also for slime production using Congo Red Agar. Both tested ionophore antibiotics showed the highest activity against planktonic bacteria of clinical as well as standard S. epidermidis strains and effectively inhibited the formation of bacterial biofilm.

Looking for new preparations for antibacterial therapy. IV. New antimicrobial agents from the aminoglycoside, macrolide and tetracycline groups in clinical trials.

This paper is the fourth in a series on the search for new antibacterial therapies, and covers new compounds belonging to the aminoglycoside, macrolide and tetracycline groups of antibiotics. The article describes eight new substances at the clinical trial stage of development. One of them is an aminoglycoside (plazomicin), four are macrolides, collectively known as ketolides (cethromycin, solithromycin, EDP-420 and EDP-788), and the remaining three are members of the tetracycline group (omadacycline, eravacycline, sarecycline). Despite the long-term and very expensive process of collecting documentation proving the efficacy of antimicrobial drugs, there is a possibility, that particular compounds find use as active ingredients of medicinal products allowing for the triumph over the clinically relevant, dangerous bacteria.

Evaluation of mycobactericidal activity of selected chemical disinfectants and antiseptics according to European standards.

BACKGROUND: The history of the investigation of standardized mycobactericidal activity of disinfectants and antiseptics is not very long. There is growing interest among the manufacturers of disinfectants in carrying out research on the antimicrobial activities in accordance with European standards (EN). This research could facilitate the introduction of high-quality disinfectants to the market. The aim of this study was to evaluate the mycobactericidal activity of selected chemical disinfectants and antiseptics used in the medical and veterinary fields. MATERIAL AND METHODS: This study included 19 products submitted to the National Medicines Institute in Poland for evaluation of mycobactericidal activity. These products contain in their composition active substances belonging to different chemical groups, including aldehydes, alcohols, amines, quaternary ammonium compounds, phenols, guanidine, and oxidizing compounds. This study, conducted according to the manufacturers' description of the preparations, was carried out in accordance with European standards, which also met the Polish standards: PN-EN 14204: 2013, PN-EN 14348: 2006, and PN-EN 14563: 2012. RESULTS: Tested products for disinfection and antiseptics containing active substances from different chemical groups showed high mycobactericidal activity and met the requirements of the appropriate European standards in most cases. In the case of products containing guanidine and amine compounds, the concentration of active ingredients used in the test and the test conditions specified by the manufacturer did not provide the mycobactericidal activity required by the standards. CONCLUSIONS: Prior to the launch of a new product on the market, it is important to establish the appropriate usage and testing conditions of the preparation, such as its practical concentration, contact time, and environment condition (clean or dirty).

Recent development of potent analogues of oxazolidinone antibacterial agents.

The oxazolidinones are a new and potent class of antimicrobial agents with activity mainly against Gram-positive strains. The commercial success of linezolid, the only FDA-approved oxazolidinone, has prompted many pharmaceutical companies to devote resources to this area of investigation. Until now, four types of chemical modifications of linezolid and oxazolidinone-type antibacterial agents, including modification on each of the A-(oxazolidinone), B-(phenyl), and C-(morpholine) rings as well as the C-5 side chain of the A-ring substructure, have been described. Division into sections according to side chain modification or the type of ring will be used throughout this review, although the process of synthesis usually involves the simultaneous modification of several elements of the linezolid substructure; therefore, assignment into the appropriate section depends on the structure-activity relationships (SAR) studies. This review makes an attempt to summarise the work carried out in the period from 2006 until mid-2012.

Antibacterial activity of selected commercial products for mouth washing and disinfection, assessed in accordance with PN-EN 1040.

BACKGROUND: Currently, there is a wide range of products for mouth washing on the Polish market. They have different qualitative and quantitative compositions, and they differ particularly in the concentration of active substances. In antisepsis and disinfection, the significant reduction in number of cells of microorganisms in a particular environment is very crucial. The chemical agents should provide a significant decrease in number of microorganisms in a relatively short time. The purpose of this study was to examine the bactericidal activity of selected herbal products used for treatment of inflammation, and disinfection and washing of the mouth, having antibacterial activity as declared by the manufacturers. MATERIAL AND METHODS: The study included 28 products for mouth washing and disinfection available in Poland. Bactericidal activity was studied using a quantitative suspension test according to the standard PN-EN 1040. RESULTS: Only 1 of 4 tested herbal products, registered as medicinal products, showed satisfactory antibacterial activity when they were used according to the manufacturer's recommendations. A total of 13 preparations (48%) complied with the standard requirements against all tested strains. Up to 19% of products showed no bactericidal activity against bacterial strains, and up to 33% were only effective against certain microorganisms. CONCLUSIONS: The informational literature accompanying most antiseptics should be corrected by the manufacturers, providing information about antimicrobial activity consistent with the requirements of applicable standards. The information on the packaging or in the leaflets for antiseptic products should be corrected by the manufacturers to include accurate information on antimicrobial activity.

Biological evaluation of quaternary bis ammonium salt and cetylpyridinum bromide against S. epidermidis biofilm.

Quaternary ammonium compounds are broad-spectrum bacteriocides widely used as antiseptics, disinfection and preservation agents. The aim of this study was to examine the activity of two quaternary ammonium salts, cetylpyridinum bromide and a newly synthesized quaternary bis ammonium salt, against S. epidermidis biofilm. The average values of killing efficiency for cetylpyridinum bromide ranged from 26.6% to 64.1% for all tested concentrations (0.125 to 8.0 microg x mL(-1)) and for quaternary bis ammonium salt the percentage of killing efficiency ranged from 59.7% to 88.4% for tested concentrations (from 2.0 to 128.0 microg x mL(-1)). Both tested compounds significantly affect staphylococcal biofilms, but any of used concentrations caused a total eradication of bacterial biofilm.

[Evaluation of biocidal properties of silver nanoparticles against cariogenic bacteria]. / Ocena bakteriobójczej aktywnosci koloidalnego roztworu nanoczastek srebra w stosunku do bakterii próchnicotwórczych.

INTRODUCTION: Antimicrobial properties of silver nanoparticles (SNP's) have been recentl well evaluated, and now are being considered as excellent candidates for therapeutic purposes. It is confirmed, that various solutions of colloidal SNP's possess significant antibacterial properties against such species as: Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa even at low concentrations, although there have been so far only a few researches evaluating antimicrobial activity of SNP's against cariogenic bacteria: Streptococcus mutans, Streptococcus salivarius and Streptococcus mitis responsible for initiation of dental carries. Tooth decay is infectious disease an worldwide, which may occur in patients of every age. Nanotechnology creates a new approach of designing of medical devices preventing or reducing bacterial colonization. METHODS: Colloidal silver solution (CSS) of concentration 350 ppm was used in this research. Nanoparticles size, shape and solution stability were evaluated. 16 strains of cariogenic bacteria, 4 isolates of each species: S. mutans, S. salivarius, S. sanguinis and S, mitis were obtained from plaque swabs of 7 patients treated for dental carries at Department of Conservative Dentistry, Medical University of Warsaw. MIC and MBC values for CSS's were evaluated. RESULTS: CSS used in this research is of good stability. No agglomeration or coalescence was observed during 24 hours of experiment. Silver nanoparticles were of round shape and had mean size of 67 nm. MIC values were: 12-25 ppm for S. salivarius, 25 ppm for S. sanguinis, 50-100 ppm for S. mitis and 50 ppm for S. mutans, while MBC values after 1 hour of bacterial contact with nanoparticles were 200-350 ppm for all cariogenic bacterial species. After 24 hours of contact MBC values were: 25-50 ppm for S. salivarius and S. sanguinis, 100-200 ppm for S. mitis and 200 ppmfor S. mutans. CONCLUSIONS: Antimicrobial properties of CSS depend on nanoparticles concentration and interaction time with bacteria. The susceptibility of cariogenic oral streptococci to silver nanoparticles is diversified. Sufficient concentration which inhibited all cariogenic bacteria in our research was 200 ppm after long (24 hours) period of silver nanoparticles interaction with bacteria.

[Microbiological characteristics of selected liquid soaps for hands washing]. / Mikrobiologiczna charakterystyka wybranych plynnych produktów stosowanych do mycia rak.

INTRODUCTION: According to common belief, supported by the authority of the World Health Organization - WHO, the common (social) hand washing is the simplest, cheapest and the most effective way of reduction the hospital-acquired infections. For this purpose products of"liquid soaps", present in a large number on the market, are most often applied. Microbiological status (microbiological purity and antimicrobial activity) of"liquid soaps" available on the Polish market is not known, because relevant routinely studies have not been performed. Only the antibacterial and / or antifungal activity of certain formulations is sometimes assessed, especially when the manufacturer suggests the standardized application of the products for surgical or hygienic procedures. The aim of this study was to determine the microbiological quality, especially microbiological purity and antimicrobial activity of the selected hands washing products, presents on the Polish market. METHODS: The 12 selected commercial products, available on the market in Poland, dedicated for hands washing were included into study. Microbiological purity test was carried out in accordance with the Polish Pharmacopoeia (FP) monograph (FP monograph numbers correspond to numbers of the European Pharmacopoeia monograph- Ph. Eur.) No 2.6.12 "Microbiological examination of non-sterile products: microbial enumaration tests", and the monograph of FP No. 2.6.13 "Microbiological examination of non-sterile products: test for specified microorganisms". The following physico-chemical properties of soaps were examined: the pH of the formulations was measured according to the monograph FP No. 2.2.3. "Potentiometric determination of pH", the density of products was assayed according to the monograph FPNo. 2.2.5. "Relative density" and determination the water activity was performed by monograph FP No 2.9.39 "Water-solid interactions: determination of sorption-desorption isotherms and of water activity". Next, antibacterial and antifungal protection was determined in accordance with the monograph FP No 5.1.3. "Efficacy of antimicrobial preservation". The study of antimicrobial activity was carried out in accordance with PN-EN 1040 "Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of basic bactericidal activity of chemical disinfectants and antiseptics - Test method and requirements (phase 1)". Finally, using the "time-kill" method the survival of microorganisms after different contact times of the products with bacteria and fungi were determined. RESULTS: All the examined products showed a very high microbiological purity. None of the formulations was characterized by a high acidity or alkalinity. All the analyzed products were slightly thicker than water, but such density of the preparation does not seem to be important parameter in the growth of microorganisms. The results of water activity estimation - the parameter indicating the presence of free, not chemically bound water stimulating microbes growth - do not show that low water content in the preparation may inhibit bacteria and fungi growth. Taking into consideration the antimicrobial protection of the products demonstrated in the tests carried out in accordance within FP monograph No 5.1.3. and PN-EN 1040, and analysing curves indicating killing rate of bacteria and fungi obtained by "time-kill" method, the microorganisms contaminating the products generally should not multiply in their environment, and gradually they die - what can take many hours or even days. CONCLUSIONS: The cases of bacterial infections connected with the usage of non-medical liguid soaps, applied in the health care units and described in the literature, should be considered as related rather to contamination of plastic packaging and dosage system, then to contamination of preparation itself inside the package. It was proved, that in all tested products amount of contaminating microbes diminishes in time. The dynamics of this process depends on the microorganisms character - bacteria dies quicker then fungi. The special attention should be given to washing, cleaning and disinfection of preparation dispensing systems, to avoid microbial contamination of product doses applied directly on the hands. It should be emphasized that only formulations containing antimicrobial agents in an appropriate amount, eliminate microorganisms from the skin surface fast and effectively. In case of hygienic and surgical procedures following the standardized manner in order to obtain required reduction rate of microorganisms in a short time - only products complying with appropriate EN standards are suitable. For these puroposes, the popular "liquid soaps" should not be used.

Looking for the new preparations for antibacterial therapy III. New antimicrobial agents from the quinolones group in clinical trials.

There is an essential need for searching for the new compounds effective in the treatment of infections caused by multidrug-resistant bacteria. This paper is the third part of a series associated with the exploration of new antibacterial agents and it discusses the compounds belonging to the group of quinolones and substances possessing a hybrid structure composed of the quinolone molecule and other compounds. Eleven new substances at the stage of clinical trials are presented. Three of them belong to the group of non-fluorinated quinolone (nemonoxacin, ozenoxacin and KRP-AM 1977X), while six are the quinolones containing fluorine atom at 6 position of the carbon atom in the quinoline ring (zabofloxacin, finafloxacin, delafloxacin, JNJ-Q2, WCK771 and KPI-10). The remaining two compounds possess a hybrid construction composed of the quinolone structure and other molecules (cadazolid and CBR-2092). There is a chance in the near future, that the presented compounds can extend the range of existing antibacterial drugs and provide an alternative to currently available medicinal products.

Looking for the new preparations for antibacterial therapy. II. Clinical trials; new beta-lactam antibiotics and beta-lactamase inhibitors.

To obtain a status of a medicinal product, a compound possessing potential antimicrobial activity and displaying no cytotoxicity, must undergo three phases of clinical trials to prove its therapeutic efficacy, safety and quality. Properties of the compound should be based on the results of studies meeting specific criteria. Studies should be: randomized, double-blind, involving sufficient number of volunteers, concerning the infections localized in strictly defined area and caused by identified microorganisms. After the medicinal product is authorized to be on the market, clinical trials of the fourth phase are carried out to detect adverse effects, overdose symptoms, interactions of the new drug with other medicinal products and to establish characteristic of activity among groups such as children, elderly, women in pregnancy and patients suffering from other diseases, but only if the benefits of receiving treatment outweigh the risks. This article is a second part of the series associated with searching for new antibacterial agents and it relates to performance of clinical trials and the new compounds belonging to the class of beta-lactams. Among the 9 presented compounds, candidates to become medicinal products, two belong to the cephalosporins (CXA-101, S-649266), one to carbapenems (razupenem), three to monobactams (BAL30072, BAL30376, MC-1) and three to beta-lactamase inhibitors (NXL-104, MK-7655, ME1071).

Reduction of the neutralisation time during antimicrobial activity testing of disinfectants according to European Standards.

BACKGROUND: Evaluation of the biocidal activity of chemical disinfectants and antiseptics according to European Standards (EN) is based on determination of the reduction of the number of viable test microorganisms under defined conditions. OBJECTIVE: The objective of this study was to investigate whether reducing the neutralization time required following declared product contact times for the tested microorganisms yields method validations. MATERIAL AND METHODS: This study was conducted on 14 products containing active substances from different chemical groups: alcohols, aldehydes, biguanides, quaternary ammonium compounds, phenols, amines derivatives, oxidizing agents. These products were tested according to phase 1 tests: EN 1040:2005 and EN 1275:2005 and then according to phase 2, step 1 tests: Draft EN 13727:2005 and EN 13624:2003. Biocidal activity was evaluated using the following test organisms: S. aureus ATCC 6538, P. aeruginosa ATCC 15442, E. coli NCTC 10538, E. coli ATCC 10536, E. hirae ATCC 10541, C. albicans ATCC 10231 and A. brasiliensis ATCC 16404. RESULTS: Validation C results for all products and tested microorganism strains were at least half of the density of the suspension for validation (Nvo) after only 10 s of neutralization. Furthermore, results from test procedures performed in parallel were also positive except 5 products toward A. brasiliensis. CONCLUSIONS: The results of our study confirm that the contact time described in the European Standards phase 1: EN 1040 and EN 1275, as well as phase 2, step 1: Draft EN 13727 and EN 13624 can be precisely determined in spite of reducing the neutralization time from 5 minutes to even 10 seconds.

Do NSAIDs and Other Pain Relief Drugs Can Inhibit the Growth of Lactobacillaceae?

Non-steroidal anti-inflammatory drugs (NSAIDs) commonly used in clinical practice may cause gastrointestinal injuries and influence the gut microbiota. This study investigated the effects of various NSAIDs and some analgesics on the viability of Lactobacillaceae strains (including probiotic strains) in vitro. It was found that diclofenac, ibuprofen, ketoprofen, dexketoprofen, flurbiprofen, and acetylsalicylic acid inhibited the growth of lactobacilli at a concentration of 0.05-3.2 mg/ml. These MICs of NSAIDs are well above therapeutic plasma concentrations achieved in humans, indicating that the tested drugs should not inhibit the growth of lactobacilli in the human digestive tract.