scholarly journals Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for Twelve Antimicrobials (Biocides and Antibiotics) in Eight Strains of Listeria monocytogenes

Biology ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 46
Author(s):  
Cristina Rodríguez-Melcón ◽  
Carlos Alonso-Calleja ◽  
Camino García-Fernández ◽  
Javier Carballo ◽  
Rosa Capita

When selecting effective doses of antimicrobials, be they biocides or antibiotics, it is essential to know the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of these substances. The present research determined the MICs and MBCs for three biocides, sodium hypochlorite (SH), benzalkonium chloride (BC), and peracetic acid (PAA), and nine antibiotics in eight strains of Listeria monocytogenes of varying serotypes. Marked intra-species differences were observed in the resistance of L. monocytogenes to the biocides and antibiotics. The MICs (ppm) for the biocides ranged between 1750 and 4500 for SH, 0.25 and 20.00 for BC, and 1050 and 1700 for PAA. Their MBCs (ppm) ranged from 2250 to 4500 for SH, 0.50 to 20.00 for BC, and 1150 to 1800 for PAA. The MICs (ppm) for antibiotics lay between 1 and 15 for ampicillin, 8 and 150 for cephalothin, 20 and 170 for cefoxitin, 0.05 and 0.20 for erythromycin, 4 and 50 for chloramphenicol, 3 and 100 for gentamicin, 2 and 15 for tetracycline, 2 and 80 for vancomycin, and 160 and 430 for fosfomycin. The corresponding MBCs (ppm) were from 5 to 20 for ampicillin, 9 to 160 for cephalothin, 70 to 200 for cefoxitin, 4 to 5 for erythromycin, 9 to 70 for chloramphenicol, 5 to 100 for gentamicin, 3 to 30 for tetracycline, 3 to 90 for vancomycin, and 160 to 450 for fosfomycin. Notably, erythromycin showed considerable efficacy, demonstrated by the low values for both MIC and MBC. Based on EUCAST and the CLSI criteria, all strains were susceptible to erythromycin. All strains were resistant to cephalothin, cefoxitin, gentamicin, and fosfomycin. Further values for resistance were 87.50% for ampicillin and vancomycin, 75.00% for tetracycline, and 62.50% for chloramphenicol. The high prevalence of antibiotic resistance is a matter for concern. A positive correlation was found between MIC and MBC values for most of the biocides and antibiotics. The higher the hydrophobicity of the cell surface, the higher the susceptibility to biocides, suggesting that surface characteristics of bacterial cells influence resistance to these compounds.

2020 ◽  
Vol 8 (2) ◽  
pp. 184
Author(s):  
Matthias Noll ◽  
Katharina Trunzer ◽  
Antje Vondran ◽  
Szilvia Vincze ◽  
Ralf Dieckmann ◽  
...  

The objective of our study was to investigate the effects of benzalkonium chloride (BC) adaptation of L. monocytogenes on the susceptibility to antimicrobial agents and on the viable but non culturable (VBNC) state of the bacterial cells. We adapted L. monocytogenes SLCC2540 to BC by applying BC below minimum inhibitory concentration (MIC) to above minimum bactericidal concentration (MBC). The culturable fractions and the susceptibility of adapted and parental cells to BC were assessed. In addition, cell membrane permeability and glucose uptake were analyzed by multi parametric flow cytometry using the fluorescent agents SYTO9, propidium iodide, and 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D-glucose (2-NBDG). Adapted cells displayed a two-fold MIC increase of BC and reduced antibiotic susceptibility. At high BC concentrations, the decrease in the number of colony forming units was significantly lower in the population of adapted cells compared to parental cells. At the same time, the number of metabolically active cells with intact membranes was significantly higher than the number of culturable cells. Growth-independent viability assays revealed an adapted subpopulation after BC application that was not culturable, indicating increased abundance of viable but nonculturable (VBNC) cells. Moreover, adapted cells can outcompete non-adapted cells under sublethal concentrations of disinfectants, which may lead to novel public health risks.


Infectio ◽  
2017 ◽  
Vol 21 (4) ◽  
Author(s):  
Daniel Felipe Vásquez-Giraldo ◽  
Gerardo Andrés Libreros-Zúñiga ◽  
María Del Pilar Crespo-Ortiz

Background: Bacterial responses to biocide exposure and its effects on survival and persistence remain to be studied in greater detail.Aim: To analyse the viability and survival of environmental isolates from household and hospital settings after biocide exposure.Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of chlorhexidine (CHxG), benzalkonium chloride (BAC) and triclosan (TC) were determined in isolates of Pseudomonas aeruginosa, Acinetobacter baumannii complex and Escherichia coli collected from hospital and households environments. Viability was monitored after exposure and removal of biocides using agar cultures and flow cytometry.Findings: P. aeruginosa isolates showed greater tolerance for all biocides tested whereas A. baumannii complex and E. coli were less tolerant.When compared with reference strains, biocide tolerance was up to 8 to 13-fold higher for TC and BAC respectively. Flow cytometry showed that biocide exposure may induce viable but non-growing states in P. aeruginosa and E. coli isolates before becoming fully replicative. Changes in the susceptibility profile in one isolate of A. baumannii complex were observed after biocide exposure.Discussion: Bacteria isolates from hospital and households were able to recover after biocide exposure at bactericidal concentrations favouring persistence and spread of biocide-tolerant strains. This study reinforces that cleaning compliance should be monitored by non-culture based tests. Novel formulations in cleaning and disinfection protocols should be revisited in hospitals harbouring P. aeruginosa and A. baumannii multidrug resistant isolates.


2020 ◽  
Vol 10 (3) ◽  
pp. 988 ◽  
Author(s):  
Lourenço Bonneville ◽  
Sagrario Ortiz ◽  
Vera Maia ◽  
Luisa Brito ◽  
Joaquín V. Martínez-Suárez

Listeria monocytogenes is one of the main foodborne pathogens. The formation of biofilms by L. monocytogenes contributes to its resistance to disinfectants, which represents a serious risk for food production plants. The aim of this study was to compare the effect of sub-inhibitory concentrations of benzalkonium chloride (BAC) (1.25 or 2.5 mg/L) on biofilm production and on biofilm reduction after exposure to an inhibitory concentration of BAC (1280 mg/L) in two isogenic L. monocytogenes strains: the BAC-sensitive wild-type strain S2-1 and its BAC-resistant mutant derivative S2BAC, which presented a multidrug resistance phenotype. The biofilm-forming ability of the strains under different BAC concentrations was evaluated by the resazurin method using polystyrene microplates. The biofilm reduction after BAC exposure was evaluated by using stainless steel coupons (SSCs). When the resazurin method was used, S2BAC produced significantly more biofilm in the presence of a sub-inhibitory concentration of BAC compared to that in the culture medium without BAC (p < 0.05). When the SSC method was used, the presence of sub-inhibitory concentrations of BAC resulted in a higher resistance of the biofilm for S2BAC compared to that in the culture medium without BAC (p < 0.05). This was not observed with the sensitive S2-1 strain. These results suggest that biofilm behavior depends on the strain and sub-inhibitory concentrations of disinfectants and may explain the ability of certain isolates to persist in niches of food processing plants.


2011 ◽  
Vol 74 (6) ◽  
pp. 1017-1021 ◽  
Author(s):  
I. SAKARIDIS ◽  
N. SOULTOS ◽  
E. IOSSIFIDOU ◽  
A. PAPA ◽  
I. AMBROSIADIS ◽  
...  

This study was conducted to determine the prevalence and antimicrobial resistance of Listeria monocytogenes recovered from chicken carcasses in slaughterhouses in Northern Greece. A total of 100 poultry samples (300 carcasses) were examined for Listeria spp. The samples were neck skin taken from four different slaughterhouses in Northern Greece. Forty samples were also taken from the environment of the slaughterhouses. Identification of L. monocytogenes was carried out by PCR and fingerprinting of the isolates by random amplified polymorphic DNA. L. monocytogenes strains isolated from chicken carcasses and from the environment of the slaughterhouses were also examined for antibiotic resistance. Fifty-five isolates of L. monocytogenes were tested for susceptibility to 20 antibiotics using the disk diffusion method. Listeria spp. were present in 99 of the poultry samples tested (99%), and 38 yielded L monocytogenes (38%). L. monocytogenes was also isolated in 80% of samples from the environment of a certain slaughterhouse, while the other slaughterhouses were found to be contaminated only with Listeria spp. All isolates were resistant to nalidixic acid and oxolinic acid, the majority of them to clindamycin, and only a few to tetracycline and oxytetracycline, whereas they were found to be susceptible to all other antimicrobials. The results of this study demonstrate a high prevalence of L. monocytogenes contamination in chicken carcasses, and all isolates were found to be sensitive to the antimicrobials most commonly used to treat human listeriosis.


2020 ◽  
Vol 17 (10) ◽  
pp. 1104-1112
Author(s):  
Kamol YUSOOK ◽  
Pettaya PANVONGSA

Lupinifolin from Derris reticulata Craib. was extracted with hexane by Soxhlet extractor and purified by crystallization. The yellow needle-shaped lupinifolin crystals were identified and confirmed by nuclear magnetic resonance (NMR) spectra and Liquid chromatography mass spectrometry (LC/MS). The lupinifolin showed minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 8 and 16 µg/ml against Methicillin resistant S. aureus (MRSA), respectively. The flow cytometry (FCM) was performed to determine the alteration of cytoplasmic membrane (CM) permeability of MRSA by using Propidium iodide (PI) 5 µg/ml as an indicator for bacterial membrane integrity. It was found that the bacterial CM permeability was effected by lupinifolin with the MIC of 8 µg/ml comparable to the control when investigated by Propidium iodide intensity. Additionally, DNA laddering assay was carried out to evaluate apoptosis in bacterial cells. It was shown that the lupinifolin has no effect on DNA fragmentation.


2011 ◽  
Vol 13 (3) ◽  
pp. 293-297 ◽  
Author(s):  
C.E Castro ◽  
J.M Ribeiro ◽  
T.T Diniz ◽  
A.C Almeida ◽  
L.C Ferreira ◽  
...  

The antibacterial effect of Lippia sidoides (rosemary pepper) essential oil was tested against the bacteria Staphylococcus aureus and Escherichia coli isolated from homemade Minas cheese produced in Brazil. The Minimum Inhibitory Concentration (MIC) determined in the Dilution Test was 13 µL oil mL-1 for both bacteria, which characterizes inhibitory action in broth for a 24-hour interaction period. The Minimum Bactericidal Concentration (MBC) determined in the Suspension Test, with one minute of contact, was 25 µL oil mL-1 for both tested bacteria, obtaining at this concentration a bactericidal effect of 99.9% on the viable bacterial cells from each sample. Results demonstrated the bacterial activity of Lippia sidoides essential oil against S. aureus and E. coli, suggesting its use as an antibacterial agent in foods.


AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Moj Khaleghi ◽  
Sadegh Khorrami

AbstractConsidering the prevalence of resistance to antibiotics, the discovery of effective agents against resistant pathogens is of extreme urgency. Herein, 26 mecA-positive methicillin-resistant S. aureus (MRSA) isolated from clinical samples were identified, and their resistance to 11 antibiotics was investigated. Next, the antibacterial and anti-biofilm activity of the ethanolic extract of M. communis on these strains was evaluated. Furthermore, the effect of this extract on the expression of biofilm-associated genes, icaA, icaD, bap, sarA, and agr, was studied. According to the results, all isolated strains were multidrug-resistant and showed resistance to oxacillin and tetracycline. Also, 96.15 and 88.46 % of them were resistant to gentamicin and erythromycin. However, the extract could effectively combat the strains. The minimum inhibitory concentration (MIC) against different strains ranged from 1.56 to 25 mg/ml and the minimum bactericidal concentration (MBC) was between 3.125 and 50 mg/ml. Even though most MRSA (67 %) strongly produced biofilm, the sub-MIC concentration of the extract destroyed the pre-formed biofilm and affected the bacterial cells inside the biofilm. It could also inhibit biofilm development by significantly decreasing the expression of icaA, icaD, sarA and bap genes involved in biofilm formation and development. In conclusion, the extract inhibits biofilm formation, ruins pre-formed biofilm, and kills cells living inside the biofilm. Furthermore, it down-regulates the expression of necessary genes and nips the biofilm formation in the bud.


2020 ◽  
pp. 514-524
Author(s):  
Omnia A Khalil ◽  
Mona I Enbaawy ◽  
Taher Salah ◽  
Hossam Mahmoud ◽  
Eman Ragab

Despite the presence of modern antibacterial drugs, bacterial infections are still a major threatening problem due to the enormous increase in multi-drug-resistant bacteria. Nanoparticles have been extensively used as an applicable and safe alternative to antibiotics. The present study aimed to explore the inhibitory effect of silver nanoparticles on Extended Spectrum Beta lactamase (ESBL) producing E. coli and Klebsiella spp. in vitro as well as their effect on the expression of antibiotic resistance genes. Different samples (i.e., wound swabs, Fecal swabs, and urine samples) were collected from dogs and cats. Phenotypic and molecular identification, antibiotic susceptibility testing, and double-disk synergy test were carried out for the identification of ESBL producing E. coli and Klebsiella spp. Silver nanoparticles were tested for their in vitro antibacterial potential and there were reports of their minimum inhibitory concentration and minimum bactericidal concentration. Moreover, the effect of silver nanoparticles on the expression of antibiotic resistance genes (i.e., blaTEM, blaSHV, and blaCTX) was assessed as well as their effect on the structural integrity of the bacterial cells using Scanning Electron Microscope (SEM). Results revealed that 23 isolates (19.16%) (E. coli=17, Klebsiella spp.=6) were confirmed as ESBL producing. Silver nanoparticles indicated a promising antibacterial effect where the minimum inhibitory concentration of AgNPs for ESBL producing E. coli was measured as 0.31 mg/ml, and 0.62 mg/ml for ESBL-producing Klebsiella spp., while the minimum bactericidal concentration of ESBL-producing E. coli and Klebsiella spp. was reported as 0.15 mg/ml and 0.3 mg/ml, respectively. Consequently, the expression of antibiotic resistance genes was downregulated in both bacteria species and there was a noticeable toxic effect of AgNPs on E. coli and Klebsiella spp. cells which was investigated using SEM. It can be concluded that silver nanoparticles have a promising antibacterial activity and could be considered an applicable alternative for the control of ESBL producing bacteria.


Author(s):  
R. H. Liss

Piperacillip (PIP) is b-[D(-)-α-(4-ethy1-2,3-dioxo-l-piperzinylcar-bonylamino)-α-phenylacetamido]-penicillanate. The broad spectrum semisynthetic β-lactam antibiotic is believed to effect bactericidal activity through its affinity for penicillin-binding proteins (PBPs), enzymes on the bacterial cytoplasmic membrane that control elongation and septation during cell growth and division. The purpose of this study was to correlate penetration and binding of 14C-PIP in bacterial cells with drug-induced lethal changes assessed by microscopic, microbiologic and biochemical methods.The bacteria used were clinical isolates of Escherichia coli and Pseudomonas aeruginosa (Figure 1). Sensitivity to the drug was determined by serial tube dilution in Trypticase Soy Broth (BBL) at an inoculum of 104 organisms/ml; the minimum inhibitory concentration of piperacillin for both bacteria was 1 μg/ml. To assess drug binding to PBPs, the bacteria were incubated with 14C-PIP (5 μg/0.09 μCi/ml); controls, in drug-free medium.


2020 ◽  
Vol 21 (11) ◽  
pp. 1129-1137 ◽  
Author(s):  
Somayeh Mirsadeghi ◽  
Masoumeh F. Koudehi ◽  
Hamid R. Rajabi ◽  
Seied M. Pourmortazavi

Background: Herein, we report the biosynthesis procedure to prepare silver nanoparticles as reduction and capping agents with the aqueous plant extract of Perovskia abrotanoides. Methods: The therapeutic application of silver nanoparticles entirely depends on the size and shape of the nanoparticles therefore, their control during the synthesis procedure is so important. The effects of synthesis factors, for example, silver ion concentration, the mass of plant extract, reaction time and extraction temperature, on the size of silver particles were considered and optimized. Several analytical methods were used for the characterization of silver NPs including FT-IR and UV–Vis spectrophotometer, XRD and SEM. Results: The results showed that the mean size of the silver particles was about 51 nm. Moreover, the antibacterial properties of biosynthesized silver NPs were investigated by the minimum inhibitory concentration, minimum bactericidal concentration, and Well-diffusion tests. The minimum inhibitory concentration/ minimum bactericidal concentration values of silver NPs and aqueous plant extract versus Gram-positive bacteria (Staphylococcus aureus and Bacillus cereus) and Gram-negative bacteria (E. coli) were 3.03/0.00, 1.20/0.01, 3.06/0.00, 0.98/1.04, 1.00/0.05 and 1.30/0.03 (mg/mL), respectively. Conclusion: The antimicrobial activity study displayed that the synthesized silver nanoparticles by plant extract have better antimicrobial properties compared to aqueous plant extract of Perovskia abrotanoides.


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