scholarly journals Strain and Growth Conditions may Regulate Resistance of Listeria monocytogenes Biofilms to Benzalkonium Chloride

2020 ◽  
Vol 10 (3) ◽  
pp. 988 ◽  
Author(s):  
Lourenço Bonneville ◽  
Sagrario Ortiz ◽  
Vera Maia ◽  
Luisa Brito ◽  
Joaquín V. Martínez-Suárez
Keyword(s):  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Benzalkonium Chloride ◽  
Culture Medium ◽  
Wild Type Strain ◽  
Inhibitory Concentration ◽  
Resistant Mutant ◽  
Growth Conditions ◽  
Processing Plants ◽  
Mutant Derivative

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.

scholarly journals Antimicrobial Activity of Nisin and Lysozyme on Foodborne Pathogens Listeria Monocytogenes, Staphylococcus Aureus, Salmonella Typhimurium, and Escherichia Coli at Different pH

2018 ◽  
Author(s):  
Hamdollah Moshtaghi ◽  
Azadeh Rashidimehr ◽  
Behzad Shareghi
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Inhibitory Concentration ◽  
Low Ph ◽  
Bacteria Growth ◽  
Microdilution Method ◽  
Natural Preservatives

Background: To prevent or inhibit the growth of pathogenic microorganisms and food spoilage factors, many studies have been done by using natural preservatives. The aim of study was to investigate the effect of different concentrations of lysozyme and Nisin on the growth rate and also to determine the minimum inhibitory concentration (MIC) and minimum bactericidal cocentratiin (MBC) of these combinations on the bacteria of Escherichia coli, Staphylococcus aureus, Salmonella typhimorium and Listeria monocytogenes. Methods: In this study, various concentrations of lysozyme and Nisin were set in form of alone concentration and in combination concentrations (0, 19.53, 39.06, 78.13, 156.25, 312.5, 625, 1250, 2500, 5000) in vitro conditions and 6 pH 5.5, 6, 6.5, 7, 7.5, and 8. Microdilution method at 24°C was done and the combined effect on bacteria growth was read by using ELISA reader. Results: The results showed that lysozyme was less effective on Escherichia coli and Nisin was less effective on Listeria monocytogenes. Moreover, combining lysozyme and Nisin at low pH decreased the MIC. Conclusions: The results of the study showed that the effect of combining lysozyme and Nisin on Staphylococcus aureus is above all other bacteria and at low pH reduces the MIC.

scholarly journals Biocide-Tolerant Listeria monocytogenes Isolates from German Food Production Plants Do Not Show Cross-Resistance to Clinically Relevant Antibiotics

2019 ◽  
Vol 85 (20) ◽  
Author(s):  
A. Roedel ◽  
R. Dieckmann ◽  
H. Brendebach ◽  
J. A. Hammerl ◽  
S. Kleta ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Food Production ◽  
Cross Resistance ◽  
High Genetic Diversity ◽  
Content Type ◽  
Processing Plants ◽  
German Food

ABSTRACT Contamination of food during processing is recognized as a main transmission route of Listeria monocytogenes. To prevent microbial contamination, biocides are widely applied as disinfectants in food processing plants. However, there are concerns about the development of antimicrobial resistance in foodborne pathogens due to widespread biocide usage. In our study, 93 L. monocytogenes isolates from German food production facilities were (i) tested for biocide and antibiotic susceptibility using broth microdilution assays, (ii) analyzed for links between reduced biocide susceptibility and antibiotic resistance, and (iii) characterized by whole-genome sequencing, including the detection of genes coding for biocide tolerance, antibiotic resistance, and other virulence factors. Fifteen L. monocytogenes isolates were tolerant to benzalkonium chloride (BAC), and genes conferring BAC tolerance were found in 13 of them. Antibiotic resistance was not associated with biocide tolerance. BAC-tolerant isolates were assigned to 6 multilocus sequence type (MLST) clonal complexes, and most of them harbored internalin A pseudogenes with premature stop codons or deletions (n = 9). Our study demonstrated a high genetic diversity among the investigated isolates including genotypes that are frequently involved in human infections. Although in vitro adaptation studies to biocides have raised concerns about increasing cross-resistance to antibiotics, our results do not provide evidence for this phenomenon in field isolates. IMPORTANCE Foodborne pathogens such as L. monocytogenes can persist in food production environments for a long time, causing perennial outbreaks. Hence, bacterial pathogens are able to survive cleaning and disinfection procedures. Accordingly, they may be repeatedly exposed to sublethal concentrations of disinfectants, which might result in bacterial adaptation to these biocides. Furthermore, antibiotic coresistance and cross-resistance are known to evolve under biocide selection pressure in vitro. Hence, antimicrobial tolerance seems to play a crucial role in the resilience and persistence of foodborne pathogens in the food chain and might reduce therapeutic options in infectious diseases.

scholarly journals Conservation and Distribution of the Benzalkonium Chloride Resistance CassettebcrABCin Listeria monocytogenes

2013 ◽  
Vol 79 (19) ◽  
pp. 6067-6074 ◽  
Author(s):  
Vikrant Dutta ◽  
Driss Elhanafi ◽  
Sophia Kathariou
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Benzalkonium Chloride ◽  
Clinical Environment ◽  
Outbreak Strain ◽  
Content Type ◽  
Chloride Resistance ◽  
Processing Plants ◽  
Hot Dog ◽  
Widespread Dissemination

ABSTRACTAnalysis of a panel of 116Listeria monocytogenesstrains of diverse serotypes and sources (clinical, environment of food processing plants, and food) revealed that all but one of the 71 benzalkonium chloride-resistant (BCr) isolates harboredbcrABC, previously identified on a large plasmid (pLM80) of the 1998-1999 hot dog outbreak strain H7858. In contrast,bcrABCwas not detected among BC-susceptible (BCs) isolates. ThebcrABCsequences were highly conserved among strains of different serotypes, but variability was noted in sequences flankingbcrABC. The majority of the BCrisolates had either the pLM80-type of organization of thebcrABCregion or appeared to harborbcrABCon the chromosome, adjacent to novel sequences. Transcription ofbcrABCwas induced by BC (10 μg/ml) in strains of different serotypes and diversebcrABCregion organization. These findings reveal widespread dissemination ofbcrABCacross BCrL. monocytogenesstrains regardless of serotype and source, while also suggesting possible mechanisms ofbcrABCdissemination acrossL. monocytogenesgenomes.

Comparison between the Resistance of Benzalkonium Chloride–Adapted and –Nonadapted Biofilms of Listeria monocytogenes to Modified Atmosphere Packaging and Nisin Once Transferred to Mussels

2011 ◽  
Vol 74 (7) ◽  
pp. 1112-1118 ◽  
Author(s):  
P. SAÁ IBUSQUIZA ◽  
J. J. R. HERRERA ◽  
M. L. CABO
Keyword(s):  
Listeria Monocytogenes ◽  
Benzalkonium Chloride ◽  
Modified Atmosphere Packaging ◽  
Inhibitory Effect ◽  
Cross Contamination ◽  
Positive Interaction ◽  
Modified Atmospheres ◽  
Processing Plants ◽  
High O2 ◽  
High Concentrations

Benzalkonium chloride–adapted and –nonadapted Listeria monocytogenes biofilm cells were transferred by contact to cooked or live mussels and packed in rich CO2 and O2, respectively. The viabilities of transferred cells during storage of these packed samples at 2.5°C were compared. In addition, in cooked mussels the combined effect of CO2 and nisin against the survival of L. monocytogenes was also studied by using a first-order factorial design. The results obtained demonstrated that biofilms formed by benzalkonium chloride–adapted L. monocytogenes cells could be more resistant to the application of modified atmospheres rich in CO2 and nisin once they have been transferred to cooked mussels by contact (simulating cross-contamination). This implies an increase in the risk associated with the presence of these cells in food processing plants. Significant empirical equations obtained after 7, 11, and 20 days showed an inhibitory effect of CO2 and nisin against L. monocytogenes. However, a significant positive interaction between both variables highlights an incompatibility between CO2 and nisin at high concentrations. Results also demonstrated that L. monocytogenes could persist after cross-contamination during the processing of live mussels, so L. monocytogenes is of concern as a contaminant in live mussels packaged in high-O2 atmospheres.

scholarly journals Heavy-Metal and Benzalkonium Chloride Resistance of Listeria monocytogenes Isolates from the Environment of Turkey-Processing Plants

2008 ◽  
Vol 74 (5) ◽  
pp. 1464-1468 ◽  
Author(s):  
S. Mullapudi ◽  
R. M. Siletzky ◽  
S. Kathariou
Keyword(s):  
Listeria Monocytogenes ◽  
Quaternary Ammonium ◽  
Food Processing ◽  
Benzalkonium Chloride ◽  
The United States ◽  
Limited Information ◽  
Chloride Resistance ◽  
Processing Plants ◽  
Serotype 1 ◽  
Serotype 4B

ABSTRACT The resistance of Listeria monocytogenes to cadmium and arsenic has been used extensively for strain subtyping. However, limited information is available on the prevalence of such resistance among isolates from the environment of food-processing plants. In addition, it is not known whether the resistance of such isolates to heavy metals may correlate with resistance to quaternary ammonium compounds extensively used as disinfectants in the food-processing industry. In this study, we characterized 192 L. monocytogenes isolates (123 putative strains) from the environment of turkey-processing plants in the United States for resistance to cadmium and arsenic and to the quaternary ammonium disinfectant benzalkonium chloride (BC). Resistance to cadmium was significantly more prevalent among strains of serotypes 1/2a (or 3a) and 1/2b (or 3b) (83% and 74%, respectively) than among strains of the serotype 4b complex (19%). Resistance to BC was encountered among 60% and 51% of the serotype 1/2a (or 3a) and 1/2b (or 3b) strains, respectively, and among 7% of the strains of the serotype 4b complex. All BC-resistant strains were also resistant to cadmium, although the reverse was not always the case. In contrast, no correlation was found between BC resistance and resistance to arsenic, which overall was low (6%). Our findings suggest that the processing environment of turkey-processing plants may constitute a reservoir for L. monocytogenes harboring resistance to cadmium and to BC and raise the possibility of common genetic elements or mechanisms mediating resistance to quaternary ammonium disinfectants and to cadmium in L. monocytogenes.

scholarly journals Listeria monocytogenes in Milk: Occurrence and Recent Advances in Methods for Inactivation

Beverages ◽  
2019 ◽  
Vol 5 (1) ◽  
pp. 14 ◽  
Author(s):  
Sarah Lee ◽  
Leandro Cappato ◽  
Jonas Guimarães ◽  
Celso Balthazar ◽  
Ramon Rocha ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Electric Fields ◽  
Culture Medium ◽  
Thermal Treatments ◽  
Important Food ◽  
Minimally Processed ◽  
Recent Advances ◽  
Processing Plants ◽  
Nutritional Qualities ◽  
Traditional Processing

Milk is one of the most important food items consumed by humans worldwide. In addition to its nutritional importance, milk is an excellent culture medium for microorganisms, which may include pathogens such as Listeria monocytogenes (L. monocytogenes). Traditional processing of milk for direct consumption is based on thermal treatments that efficiently eliminate pathogens, including pasteurization or sterilization. However, the occurrence of L. monocytogenes in milk as a consequence of failures in the pasteurization process or postpasteurization contamination is still a matter of concern. In recent years, consumer demand for minimally processed milk has increased due to the perception of better sensory and nutritional qualities of the products. This review deals with the occurrence of L. monocytogenes in milk in the last 10 years, including regulatory aspects, and recent advances in technologies for the inactivation of this pathogen in milk. The results from studies on nonthermal technologies, such as high hydrostatic pressure, pulsed electric fields, ultrasounds, and ultraviolet irradiation, are discussed, considering their potential application in milk processing plants.

Heat Resistance of Listeria monocytogenes

2001 ◽  
Vol 64 (3) ◽  
pp. 410-429 ◽  
Author(s):  
M. ELLIN DOYLE ◽  
ALEJANDRO S. MAZZOTTA ◽  
TIM WANG ◽  
DANA W. WISEMAN ◽  
VIRGINIA N. SCOTT
Keyword(s):  
Listeria Monocytogenes ◽  
Heat Resistance ◽  
Foodborne Pathogens ◽  
Fruits And Vegetables ◽  
Culture Media ◽  
Salt Content ◽  
Growth Conditions ◽  
Salmonella Spp ◽  
Different Strains ◽  
Resistance Data

The heat resistance data on Listeria monocytogenes in culture media and foods are summarized. Most heat resistance data for foods have been obtained in dairy, meat, poultry, and egg products. Limited data have been published on seafood, fruits, and vegetables. The methodologies employed have evolved over time; hence data from earlier experiments are not directly comparable to more recent studies. Many factors influence the heat resistance of L. monocytogenes. Variation exists among different strains in their ability to withstand heat treatment. In addition, heat resistance is influenced by age of the culture, growth conditions, recovery media, and characteristics of foods such as salt content, aw, acidity, and the presence of other inhibitors. Listeriae are more heat resistant than most other nonspore-forming foodborne pathogens, and thus, processing recommendations based on data from experiments with Salmonella spp. or pathogenic Escherichia coli may not be sufficient to eliminate similar numbers of L. monocytogenes. The data provided in this review may prove useful for food processors in determining appropriate times and temperatures for producing foods free of vegetative pathogens.

scholarly journals Genetic Characterization of Plasmid-Associated Benzalkonium Chloride Resistance Determinants in a Listeria monocytogenes Strain from the 1998-1999 Outbreak

2010 ◽  
Vol 76 (24) ◽  
pp. 8231-8238 ◽  
Author(s):  
Driss Elhanafi ◽  
Vikrant Dutta ◽  
Sophia Kathariou
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Benzalkonium Chloride ◽  
Genetic Basis ◽  
Genetic Characterization ◽  
Chloride Resistance ◽  
Processing Plants ◽  
Resistant Strains ◽  
Ammonium Compounds

ABSTRACT Quaternary ammonium compounds such as benzalkonium chloride (BC) are widely used as disinfectants in both food processing and medical environments. BC-resistant strains of Listeria monocytogenes have been implicated in multistate outbreaks of listeriosis and have been frequently isolated from food processing plants. However, the genetic basis for BC resistance in L. monocytogenes remains poorly understood. In this study, we have characterized a plasmid (pLM80)-associated BC resistance cassette in L. monocytogenes H7550, a strain implicated in the 1998-1999 multistate outbreak involving contaminated hot dogs. The BC resistance cassette (bcrABC) restored resistance to BC (MIC, 40 μg/ml) in a plasmid-cured derivative of H7550. All three genes of the cassette were essential for imparting BC resistance. The transcription of H7550 BC resistance genes was increased under sublethal (10 μg/ml) BC exposure and was higher at reduced temperatures (4, 8, or 25°C) than at 37°C. The level of transcription was higher at 10 μg/ml than at 20 or 40 μg/ml. In silico analysis suggested that the BC resistance cassette was harbored by an IS1216 composite transposon along with other genes whose functions are yet to be determined. The findings from this study will further our understanding of the adaptations of this organism to disinfectants such as BC and may contribute to the elucidation of possible BC resistance dissemination in L. monocytogenes.

scholarly journals Diverse Cadmium Resistance Determinants in Listeria monocytogenes Isolates from the Turkey Processing Plant Environment

2009 ◽  
Vol 76 (2) ◽  
pp. 627-630 ◽  
Author(s):  
S. Mullapudi ◽  
R. M. Siletzky ◽  
S. Kathariou
Keyword(s):  
Listeria Monocytogenes ◽  
Benzalkonium Chloride ◽  
Processing Plant ◽  
Cadmium Resistance ◽  
Resistance Determinants ◽  
Plant Environment ◽  
Processing Plants

ABSTRACT Two different cadA cadmium resistance determinants (cadA1, first identified in Tn5422, and cadA2, associated with pLM80) were detected among cadmium-resistant Listeria monocytogenes strains from turkey processing plants. Prevalence of cadA1 versus cadA2 was serotype associated. Cadmium-resistant isolates that were also resistant to benzalkonium chloride (BC) were more likely to harbor cadA2 alone or together with cadA1 than isolates that were cadmium resistant but BC susceptible.

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
Keyword(s):  
Antibiotic Resistance ◽  
Listeria Monocytogenes ◽  
Benzalkonium Chloride ◽  
Species Differences ◽  
Inhibitory Concentration ◽  
Minimum Bactericidal Concentration ◽  
Surface Characteristics ◽  
Bacterial Cells ◽  
High Prevalence ◽  
Effective Doses

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.

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