A new and efficient method to obtain benzalkonium chloride adapted cells of Listeria monocytogenes

Ibusquiza, Paula Saá; Herrera, Juan José Rodríguez; Vázquez‐Sánchez, Daniel; Parada, Adelaida; Cabo, Marta López

doi:10.1016/j.mimet.2012.07.009

Cited by 14 publications

(11 citation statements)

References 38 publications

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“…Of note, in L34-CECT324 the susceptibility of L34 toward BAC was even higher than in monospecies samples displaying a reduction value 2.35 log CFU cm –2 higher ( Table 3 ). These results are in contrast with the those of Saá Ibusquiza et al (2012b) demonstrating that in 96-h L. monocytogenes CECT 4032– P. putida CECT 911 biofilms on SS, the first displayed a five-fold increase regarding the resistance to BAC if compared with their monospecies counterparts. However, in this case, the shorter growth times used can explain the lower resistance as it has been previously published that the matureness, i.e., age, of a L. monocytogenes mixed-species biofilm is one of the determining factors affecting its overall resistance ( Rodríguez-López and Cabo, 2017 ).…”

Section: Resultscontrasting

confidence: 85%

Architectural Features and Resistance to Food-Grade Disinfectants in Listeria monocytogenes-Pseudomonas spp. Dual-Species Biofilms

2022

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Listeria monocytogenes is considered a foodborne pathogen of serious concern capable of forming multispecies biofilms with other bacterial species, such as Pseudomonas spp., adhered onto stainless steel (SS) surfaces. In an attempt to link the biofilms’ morphology and resistance to biocides, dual-species biofilms of L. monocytogenes, in co-culture with either Pseudomonas aeruginosa, Pseudomonas fluorescens, or Pseudomonas putida, were assayed to ascertain their morphological characteristics and resistance toward benzalkonium chloride (BAC) and neutral electrolyzed water (NEW). Epifluorescence microscopy analysis revealed that each dual-species biofilm was distributed differently over the SS surface and that these differences were attributable to the presence of Pseudomonas spp. Confocal laser scanning microscopy (CLSM) assays demonstrated that despite these differences in distribution, all biofilms had similar maximum thicknesses. Along with this, colocalization analyses showed a strong trend of L. monocytogenes to share location within the biofilm with all Pseudomonas assayed whilst the latter distributed throughout the surface independently of the presence of L. monocytogenes, a fact that was especially evident in those biofilms in which cell clusters were present. Finally, a modified Gompertz equation was used to fit biofilms’ BAC and NEW dose-response data. Outcomes demonstrated that L. monocytogenes was less susceptible to BAC when co-cultured with P. aeruginosa or P. fluorescens, whereas susceptibility to NEW was reduced in all three dual-species biofilms, which can be attributable to both the mechanism of action of the biocide and the architectural features of each biofilm. Therefore, the results herein provided can be used to optimize already existing and develop novel target-specific sanitation treatments based on the mechanism of action of the biocide and the biofilms’ species composition and structure.

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Section: Resultscontrasting

confidence: 85%

Architectural Features and Resistance to Food-Grade Disinfectants in Listeria monocytogenes-Pseudomonas spp. Dual-Species Biofilms

2022

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“…As a shelter for bacterial community in biofilms, EPS in the multispecies biofilms could act as a diffusionlimiting barrier against the preservatives, resulting in limited compounds access to LM in the deeper layer of the biofilms (Karygianni et al, 2020). Previous studies demonstrated a competitive interaction and increased sanitizer resistance of LM when co-cultured with Pseudomonas putida (Saa Ibusquiza et al, 2012). The dominating competitive interactions among multispecies brewery biofilm, community members confer tolerance against the antimicrobial sulphathiazole, due to a reduction in competition upon antimicrobial treatment (Parijs and Steenackers, 2018).…”

Section: Discussionmentioning

confidence: 99%

Synergism With ε-Polylysine Hydrochloride and Cinnamon Essential Oil Against Dual-Species Biofilms of Listeria monocytogenes and Pseudomonas lundensis

et al. 2022

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Various pathogenic and spoilage bacteria frequently coexist in meat processing environments and can form multispecies biofilms, causing significant health and economic issues. Despite the prevalence and coexistence, only less is known about possible interactions between Listeria monocytogenes (LM) and spoilers like Pseudomonas species, and their community-wide resistance against natural preservatives. This study evaluates the interactions between mono- or dual-species biofilms formed by LM and Pseudomonas lundensis (PL), as well as the sensitivity of these bacteria in dual-species biofilms to ε-polylysine hydrochloride (ε-PLH) alone or combined with cinnamon essential oil (CEO). The results showed that the biofilm cell density of P. lundensis in dual species was higher (p < 0.05) than LM, constituting about 85% of the total population. More biofilms and exopolysaccharide both in mono- or dual species of the two psychrotrophic strains were greatly produced at 15°C than at 30°C. The biomass, biovolume, and thickness of dual-species biofilms were significantly lower than single PL biofilm when tested using crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy, indicating the competitive interactions between them prevail. Additionally, ε-PLH significantly reduced the biofilm development as mono- and dual species in a concentration-dependent manner, especially single LM biofilm, which was consistent with the decrease in autoinducer-2 (AI-2) activity. LM as dual-species biofilms exhibited lower sensitivity to ε-PLH than its mono-biofilm probably due to protective effect conferred by PL. ε-PLH in combination with CEO, at the maximum sublethal concentrations (MSCs), showed enhanced inhibitory activity against dual-species biofilm formation, as evidenced by thin spare spatial structures and reduced AI-2 activity. In addition, the preformed dual biofilms were dramatically eradicated following treatment with ε-PLH combined with CEO at higher than minimum inhibitory concentration in comparison with either of the compounds used alone, indicating the synergistic antibiofilm of the two preservatives. This study reveals the competitive interactions between the two strains in dual-species biofilms, in which the dominant PL significantly contributed toward the tolerance of LM to ε-PLH, and the use of combined preservatives shows it is an effective strategy to control the multispecies biofilms in meat processing.

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“…Regarding isolates from food sources, L1 and L8 were isolated from RTE fish products, L34 was isolated from a frozen panga fillet in a previous study [30] and L12 was isolated from a Halibut fillet as follows: 25 g of product was mixed with 225 mL of buffered peptone water (BPW; Cultimed, Barcelona, Spain) and processed with a stomacher masticator (IUL Instruments, Barcelona, Spain) for 1 min. From this sample, serial dilutions (up to 10 3 ) were made in BPW, 1 mL of each solution was spread onto two (500 µL each) PALCAM (Liofilchem, Roseto degli Abruzzi, Italy) and incubated for 24-48 h at 37 • C.…”

Section: Origin Of L Monocytogenes Isolatesmentioning

confidence: 99%

Efficacy of Synthetic Furanones on Listeria monocytogenes Biofilm Formation

Rodríguez-López

Barrenengoa

Pascual-Sáez

et al. 2019

Foods

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Furanones are analogues of acylated homoserine lactones with proven antifouling activity in both Gram-positive and Gram-negative bacteria though the interference of various quorum sensing pathways. In an attempt to find new strategies to prevent and control Listeria monocytogenes biofilm formation on stainless steel (SS) surfaces, different concentrations of six synthetic furanones were applied on biofilms formed by strains isolated from food, environmental, and clinical sources grown onto AISI 316 SS coupons. Among the furanones tested, (Z-)-4-Bromo-5-(bromomethylene)-2(5H)-furanone and 3,4-Dichloro-2(5H)-furanone significantly (p < 0.05) reduced the adhesion capacity (>1 log CFU cm−2) in 24 h treated biofilms. Moreover, individually conducted experiments demonstrated that (Z-)-4-Bromo-5-(bromomethylene)-2(5H)-furanone was able to not only significantly (p < 0.05) prevent L. monocytogenes adhesion but also to reduce the growth rate of planktonic cells up to 48 h in a dose-dependent manner. LIVE/DEAD staining followed by epifluorescence microscopy visualisation confirmed these results show an alteration of the structure of the biofilm in furanone-treated samples. Additionally, it was demonstrated that 20 µmol L−1 of 3,4-Dichloro-2(5H)-furanone dosed at 0, 24 and 96 h was able to maintain a lower level of adhered cells (>1 log CFU cm−2; p < 0.05). Since furanones do not pose a selective pressure on bacteria, these results represent an appealing novel strategy for the prevention of L. monocytogenes biofilm grown onto SS.

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A new and efficient method to obtain benzalkonium chloride adapted cells of Listeria monocytogenes

Cited by 14 publications

References 38 publications

Architectural Features and Resistance to Food-Grade Disinfectants in Listeria monocytogenes-Pseudomonas spp. Dual-Species Biofilms

Architectural Features and Resistance to Food-Grade Disinfectants in Listeria monocytogenes-Pseudomonas spp. Dual-Species Biofilms

Synergism With ε-Polylysine Hydrochloride and Cinnamon Essential Oil Against Dual-Species Biofilms of Listeria monocytogenes and Pseudomonas lundensis

Efficacy of Synthetic Furanones on Listeria monocytogenes Biofilm Formation

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