91 research outputs found

    Effect of low doses of biocides on the susceptibility of Listeria monocytogenes and Salmonella enterica to various antibiotics of clinical importance

    Get PDF
    [EN] The use of subinhibitory concentrations of biocides in food processing environments requires special attention because it is related to potential increases in antibiotic resistance. In this study, we determined the effect of exposure to low doses of four biocides (sodium hypochlorite, SHY; benzalkonium chloride, BZK; peracetic acid, PAA; trisodium phosphate, TSP) on the resistance to 10 antibiotics and on the hydrophobicity of the cellular surface of a strain of Listeria monocytogenes serotype 1/2a (LM) and a strain of Salmonella enterica serotype Agona (SA), both of meat origin. The cultures were exposed at 37 °C in Mueller Hinton II cation-adjusted broth with 0.6% yeast extract (with 0.2% of laky horse blood added in the case of LM) to increasing concentrations of the biocides, starting with half the minimum inhibitory concentration (MIC/2) and incrementing by 1.5 times the concentration until growth was no longer observed, calculating the MIC of the antibiotics before (control cultures) and after exposure. After exposure to TSP, LM was able to grow in the presence of a concentration of the biocide 2.53 times higher than the MIC of unexposed cultures. No adaptation was observed for SHY, BZK or PAA. SA demonstrated adaptation to BZK (it tolerated a concentration 1.13 times higher than the MIC for the unadapted strain) and PAA (2.53 times). LM cultures presented increased resistance (from susceptibility to reduced susceptibility, from susceptibility to resistance, or from reduced susceptibility to resistance) to erythromycin (strains exposed to BZK, PAA and TSP) and fosfomycin (all compounds). Regarding SA, after exposure its resistance to cefoxitin (all compounds), gentamicin (all compounds), tetracycline (TSP), fosfomycin (SHY, BZK and TSP) and enrofloxacin (BZK, PAA and TSP) increased. The cell surface hydrophobicity (determined through the microbial adhesion to solvents -MATS- test) increased (LM exposed to PAA and TSP; SA exposed to BZK) or decreased (SA exposed to PAA) after contact with the biocides. These findings suggest that the use of biocides at subinhibitory concentrations can contribute to the increase in bacterial resistance to antibiotics, in addition to modifying the hydrophobicity of the cellular surface, which is related to the capacity of bacteria to form biofilm.S

    Estimation by flow cytometry of percentages of survival of Listeria monocytogenes cells treated with tetracycline, with or without prior exposure to several biocides

    Get PDF
    [EN] In certain circumstances, disinfectants are used at sublethal concentrations. The aim of this research work was to determine whether contact of Listeria monocytogenes NCTC 11994 with subinhibitory concentrations of three disinfectants widely used in food processing environments and in the health-care system, benzalkonium chloride (BZK), sodium hypochlorite (SHY) and peracetic acid (PAA), can cause the adaptation of the strain to the biocides and increase its resistance to tetracycline (TE). The minimum inhibitory concentrations (MIC; ppm) were 2.0 (BZK), 3500.0 (SHY) and 1050.0 (PAA). On exposure to increasing subinhibitory concentrations of the biocides, the maximum concentrations (ppm) of the compounds that allowed the strain to grow were (ppm) 8.5 (BZK), 3935.5 (SHY) and 1125.0 (PAA). Both the control cells (non-exposed) and the cells that had been in contact with low doses of biocides were treated with different concentrations of TE (0 ppm, 250 ppm, 500 ppm, 750 ppm, 1000 ppm and 1250 ppm) for 24, 48 and 72 h, and the survival percentages determined using flow cytometry, following dying with SYTO 9 and propidium iodide. The cells previously exposed to PAA presented higher survival percentages (P < 0.05) than the rest of the cells for most of the concentrations of TE and treatment times trialled. These results are worrying because TE is sometimes used to treat listeriosis, highlighting the importance of avoiding the use of disinfectant at subinhibitory doses. Furthermore, the findings suggest that flow cytometry is a fast and simple technique to obtain quantitative data on bacterial resistance to antibiotics.S

    Prevalence, quantification and antibiotic resistance of Listeria monocytogenes in poultry preparations

    Get PDF
    .A total of 100 samples of fresh poultry preparations were obtained from 10 retail outlets in North-Western Spain. Listeria spp. were found in 73 samples. Isolates were identified through polymerase chain reaction (PCR) as Listeria monocytogenes (56 samples), Listeria innocua (32), Listeria grayi (3), Listeria seeligeri (1) and Listeria spp. (6). In 24 samples, several different Listeria species were found. The loads of L. monocytogenes detected by quantitative polymerase chain reaction (q-PCR) in the 56 positive samples ranged from 0.05) on concentrations of L. monocytogenes. A total of 163 L. monocytogenes isolates were tested (disc diffusion) against 15 antimicrobials of clinical significance. The average number of resistances per isolate was 5.83 ± 1.64. All strains showed resistance to multiple antimicrobials (between 4 and 11). In all, 80 isolates (49.1%) showed a multi-drug resistant (MDR) phenotype, and two isolates (1.2%) showed an extensively drugresistant (XDR) phenotype. More than 50.0% of isolates showed resistance or reduced susceptibility to oxacillin, cefoxitin, cefotaxime, cefepime, rifampicin, ciprofloxacin, enrofloxacin or nitrofurantoin. This is a cause for concern because these substances are among the antibiotics used to treat human listeriosis, with rifampicin and fluoroquinolones frequently being used. The results from this research work show that poultry preparations are a potential major source of resistant L. monocytogenes strains, since these are present in some samples at high concentrations. This highlights the pressing need to handle poultry preparations correctly, so as to ensure they are sufficiently cooked and to avoid cross-contamination events.S

    Effect of sodium nitrite, nisin and lactic acid on the prevalence and antibiotic resistance patterns of Listeria monocytogenes naturally present in poultry

    Get PDF
    The impact of treating minced chicken meat with sodium nitrite (SN, 100 ppm), nisin (Ni, 10 ppm) and lactic acid (LA, 3000 ppm) on the levels of some microbial groups indicating hygiene quality were investigated. Specifically, aerobic plate counts and culture-based counts of psychrotrophic microorganisms and enterobacteria were obtained. Additionally, the prevalence of Listeria monocytogenes and the resistance of 245 isolates from this bacterium to 15 antibiotics were documented. L. monocytogenes was isolated using the ISO 11290-1:2017 method and confirmed with polymerase chain reaction using the lmo1030 gene. Antibiotic resistance was established using the disc diffusion technique (EUCAST and CLSI criteria). Twenty-four hours after treatment, the microbial load (log10 cfu/g) was reduced (p 0.05) of samples. All strains showed resistance to multiple antimicrobials (between 3 and 12). In all, 225 isolates (91.8%) showed a multi-drug resistant (MDR) phenotype, and one isolate (0.4%) showed an extensively drug-resistant (XDR) phenotype. The mean number of resistances per strain was lower (p < 0.01) in the control samples, at 5.77 ± 1.22, than in those receiving treatment, at 6.39 ± 1.51. It is suggested that the use of food additives might increase the prevalence of resistance to antibiotics in L. monocytogenes, although additional studies would be necessary to verify this finding by analyzing a higher number of samples and different foodstuffs and by increasing the number of antimicrobial compounds and concentrations to be tested.Junta de Castilla y León | Ref. LE018P20Agencia Estatal de Investigación | Ref. RTI2018-098267-R-C33Agencia Estatal de Investigación | Ref. PID2022-142329OB-C3

    Effect of low doses of biocides on the antimicrobial resistance and the biofilms of Cronobacter sakazakii and Yersinia enterocolitica

    Get PDF
    The susceptibility of Cronobacter sakazakii ATCC 29544 (CS) and Yersinia enterocolitica ATCC 9610 (YE) to sodium hypochlorite (10% of active chlorine; SHY), peracetic acid (39% solution of peracetic acid in acetic acid; PAA) and benzalkonium chloride (BZK) was tested. Minimum inhibitory concentration (MIC) values (planktonic cells; microdilution broth method) of 3,800 ppm (SHY), 1,200 ppm (PAA) and 15 ppm (BZK) for CS, and 2,500 ppm (SHY), 1,275 ppm (PAA) and 20 ppm (BZK) for YE, were found. In some instances, an increase in growth rate was observed in presence of sub-MICs (0.25MIC, 0.50MIC or 0.75MIC) of biocides relative to the samples without biocides. The cultures exhibited an acquired tolerance to biocides and an increase in antibiotic resistance after exposure to sub-MICs of such disinfectants. Strains were able to form strong biofilms on polystyrene after 48 hours (confocal laser scanning microscopy), with average biovolumes in the observation field (14,161 µm2) of 242,201.0 ± 86,570.9 µm3 (CS) and 190,184.5 ± 40,860.3 µm3 (YE). Treatment of biofilms for 10 minutes with disinfectants at 1MIC or 2MIC reduced the biovolume of live cells. PAA (YE) and BZK (CS and YE) at 1MIC did not alter the percentage of dead cells relative to non-exposed biofilms, and their effect of countering biofilm was due principally to the detachment of cells. These results suggest that doses of PAA and BZK close to MICs might lead to the dissemination of live bacteria from biofilms with consequent hazards for public healthMinisterio de Ciencia, Innovación y Universidades | Ref. RTI2018-098267-R-C33Junta de Castilla y León | Ref. LE164G1

    Relationship with Clonal Lineages and Antimicrobial Resistance

    Get PDF
    Funding: NORTE-01-0145-FEDER-030101 the projects UIDB/CVT/00772/2020 and LA/P/0059/2020 funded by the Portuguese Foundation for Science and Technology (FCT). The Ministerio de Ciencia, Innovación y Universidades (Spain, grant number RTI2018-098267-R-C33) and the Junta de Castilla y León (Consejería de Educación, Spain, grant number LE018P20). Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.This study aimed to compare the biofilm formation ability of Staphylococcus aureus isolated from a wide range of animals and study the association between biofilm formation and antimicrobial resistance and genetic lineages. A total of 214 S. aureus strains isolated from pets, livestock, and wild animals were evaluated regarding their ability to form biofilms by the microtiter biofilm assay and their structure via confocal scanning laser microscopy. Statistical analysis was used to find an association between biofilm formation and antimicrobial resistance, multidrug resistance, sequence types (STs), spa and agr-types of the isolates. The antimicrobial susceptibility of 24 h-old biofilms was assessed against minimum inhibitory concentrations (MIC) and 10× MIC of amikacin and tetracycline, and the biomass reduction was measured. The metabolic activity of biofilms after antimicrobial treatment was evaluated by the XTT assay. All isolates were had the ability to form biofilms. Yet, significant differences in biofilm biomass production were detected among animal species. Multidrug resistance had a positive association with biofilm formation as well as methicillin-resistance. Significant differences were also detected among the clonal lineages of the isolates. Both tetracycline and amikacin were able to significantly reduce the biofilm mass. However, none of the antimicrobials were able to eradicate the biofilm at the maximum concentration used. Our results provide important information on the biofilm-forming capacity of animal-adapted S. aureus isolates, which may have potential implications for the development of new biofilm-targeted therapeutics.publishersversionpublishe

    Exploring the Biofilm Formation Capacity in S. pseudintermedius and Coagulase-Negative Staphylococci Species

    Get PDF
    NORTE-01-0145-FEDER-030101 UIDB/CVT/00772/2020 LA/P/0059/2020 RTI2018-098267-R-C33 LE018P20The ability of biofilm formation seems to play an important role in the virulence of staphylococci. However, studies reporting biofilm formation of coagulase-negative staphylococci isolated from animals are still very scarce. Thus, we aimed to evaluate the biofilm-forming capacity of CoNS and S. pseudintermedius isolated from several animal species and to investigate the effect of conventional antimicrobials on biofilm reduction. A total of 35 S. pseudintermedius and 192 CoNS were included. Biofilm formation was accessed by the microtiter plate assay and the biofilms were stained by crystal violet. Association between biofilm formation and staphylococci species and antimicrobial resistance was also performed. Biofilm susceptibility testing was performed with tetracycline and amikacin at the minimum inhibitory concentration (MIC) and 10 Ă— MIC. The metabolic activity of the biofilm cells after antimicrobial treatment was accessed by the XTT assay. All isolates formed biofilm, with S. urealyticus producing the most biofilm biomass and S. pseudintermedius producing the least biomass. There was a positive association between biofilm formation and multidrug resistance as well as resistance to individual antimicrobials. Neither tetracycline nor amikacin were able to eradicate the biofilm, not even at the highest concentration used. This study provides new insights into biofilm formation and the effects of antimicrobials on CoNS species.publishersversionpublishe

    Evaluation of the phenolic profile of Castanea sativa mill. By-products and their antioxidant and antimicrobial activity against multiresistant bacteria

    Get PDF
    The chestnut industry generates a large amount of by-products. These agro-industrial wastes have been described as potential sources of phenolic compounds with high bioactive potential. Therefore, we aimed to extract the phenolic compounds from chestnut by-products and assess their antioxidant potential and evaluate their antimicrobial activity against multidrug resistant bacteria. The individual phenolic compounds in the ethanolic extracts of chestnut shell, inner shell, bur, and leaves were characterized by HPLC-DAD/electrospray ionization (ESI)-MS. The antioxidant properties were determined by DPPH and ABTS assays. The minimum inhibitory concentration (MIC) and the antimicrobial susceptibility was performed using the Kirby–Bauer disc di usion method against 10 bacterial strains. The major phenolic compounds identified in the extracts were trigalloyl-HHDP-glucose, gallic acid, quercetin, and myricetin glycoside derivatives. All chestnut by-products presented promising antioxidant activity in both assays, with leaf samples the ones presenting the highest antioxidant capacity. The inner shell’s extract was e ective against all Gram-positive and two Gram-negative bacteria; nevertheless, all extracts showed antibacterial activity. Staphylococcus epidermidis showed susceptibility to all extracts while none of the extracts was able to suppress the growth of Escherichia coli and Salmonella enteritidis. Chestnut by-products are a source of phenolic compounds with prominent antioxidant and antimicrobial activities. Nevertheless, further studies should be conducted to assess the correlation between phenolic compounds and the bioactivities obtained.info:eu-repo/semantics/publishedVersio

    Genetic characterization of methicillin-resistant staphylococcus aureus isolates from human bloodstream infections: detection of mlsb resistance

    Get PDF
    In this study we aimed to characterize antimicrobial resistance in methicillin-resistant Staphylococcus aureus (MRSA) isolated from bloodstream infections as well as the associated genetic lineages of the isolates. Sixteen MRSA isolates were recovered from bacteremia samples from inpatients between 2016 and 2019. The antimicrobial susceptibility of these isolates was tested by the Kirby–Bauer disk diffusion method against 14 antimicrobial agents. To determine the macrolide–lincosamide–streptogramin B (MLSB) resistance phenotype of the isolates, erythromycin-resistant isolates were assessed by double-disk diffusion (D-test). The resistance and virulence genes were screened by polymerase chain reaction (PCR). All isolates were characterized by multilocus sequence typing (MLST), spa typing, staphylococcal chromosomal cassette mec (SCCmec) typing, and accessory gene regulator (agr) typing. Isolates showed resistance to cefoxitin, penicillin, ciprofloxacin, erythromycin, fusidic acid, clindamycin, and aminoglycosides, confirmed by the presence of the blaZ, ermA, ermC, mphC, msrA/B, aac(6’)-Ie-aph(2’’)-Ia, and ant(4’)-Ia genes. Three isolates were Panton–Valentine-leukocidin-positive. Most strains (n = 12) presented an inducible MLSB phenotype. The isolates were ascribed to eight spa-types (t747, t002, t020, t1084, t008, t10682, t18526, and t1370) and four MLSTs (ST22, ST5, ST105, and ST8). Overall, most (n = 12) MRSA isolates had a multidrug-resistance profile with inducible MLSB phenotypes and belonged to epidemic MRSA clones.info:eu-repo/semantics/publishedVersio

    Characterization of Biofilms Formed by Foodborne Methicillin-Resistant Staphylococcus aureus

    Get PDF
    The objective of this study was to evaluate the capacity of 49 methicillin resistant Staphylococcus aureus (MRSA) from foods of animal origin (42 from dairy products and 7 from meat and meat products) to form biofilms. Overall, a higher biofilm biomass was observed for those MRSA strains harboring SCCmec type IV, while 8 MRSA strains (5 from dairy products and 3 from meat and meat products) were classified as strong biofilm formers in standard Tryptic Soy Broth medium. When a prolonged incubation period (48 h) was applied for those 8 MRSA strains, an increased biofilm biomass accumulation was observed during the time course, whereas the number of viable cells within the biofilms decreased as the biomass increased. The capacity of biofilm production correlated pretty well between the experiments using polystyrene microtiter plates and stainless steel micro-well plates, and significant higher values were observed in stainless steel when glucose was added to TSB during the enrichment. Biofilms were further characterized by confocal laser scanning microscope (CLSM), confirming that proteins and α-polysaccharides were the predominant components inside the extracellular polymeric matrix of biofilms formed by MRSA strains. In conclusion, our results confirm that MRSA isolates from foods of animal origin have significant capacity for forming biofilms with a high protein content, which can play a key role for the successful dissemination of MRSA lineages via food. Knowledge of the capacity of MRSA strains to produce biofilms, as well as characterization of the main MRSA biofilms matrix components, can help both to counteract the mechanisms involved in biofilm formation and resistance and to define more rational control strategies by using tailor-made cleaning agents
    • …
    corecore