scholarly journals Prevalence, Molecular Typing, and Determination of the Biofilm-Forming Ability of Listeria monocytogenes Serotypes from Poultry Meat and Poultry Preparations in Spain

2019 ◽  
Vol 7 (11) ◽  
pp. 529 ◽  
Author(s):  
Carlos Alonso-Calleja ◽  
Sara Gómez-Fernández ◽  
Javier Carballo ◽  
Rosa Capita
Keyword(s):  
Listeria Monocytogenes ◽  
Laser Scanning ◽  
Positive Sample ◽  
Laser Scanning Microscopy ◽  
Poultry Meat ◽  
Epidemiological Surveillance ◽  
Confocal Laser ◽  
Retail Outlets ◽  
Northwestern Spain

A study was undertaken of the presence of Listeria monocytogenes in 260 samples of poultry meat obtained from retail outlets in northwestern Spain. L. monocytogenes was detected in 20 samples (7.7%). Twenty strains (one strain per positive sample) were characterized. The strains belonged to 10 serotypes: 1/2a (2 strains), 1/2b (2), 1/2c (2), 3a (1), 3b (2), 3c (2), 4a (2), 4b (4), 4c (1), and 4d (2). Cluster analysis (ribotyping; EcoRI) showed a strong genetic relationship between strains isolated from samples coming from different outlets. Ribotyping permitted some isolates of the same serotype to be differentiated, which points to the possible usefulness of this technique in the epidemiological surveillance of L. monocytogenes. All strains formed biofilm on polystyrene, as shown by confocal laser scanning microscopy. The biovolume (between 621.7 ± 36.0 µm3 and 62,984.0 ± 14,888.2 µm3 in the observational field of 14,161 μm2), percentage of surface coverage (from 2.17 ± 0.84% to 94.43 ± 3.97%), roughness (between 0.399 ± 0.052 and 0.830 ± 0.022), and maximum thickness (between 9.00 ± 0.00 µm and 24.00 ± 14.93 µm) of biofilms varied between strains (p < 0.05). These results expand knowledge of the characteristics of L. monocytogenes isolates from poultry.

Effects of Bacteriophage P100 at Different Concentrations on the Structural Parameters of Listeria monocytogenes Biofilms

2018 ◽  
Vol 81 (12) ◽  
pp. 2040-2044 ◽  
Author(s):  
CRISTINA RODRÍGUEZ-MELCÓN ◽  
ROSA CAPITA ◽  
CAMINO GARCÍA-FERNÁNDEZ ◽  
CARLOS ALONSO-CALLEJA
Keyword(s):  
Image Analysis ◽  
Listeria Monocytogenes ◽  
Laser Scanning ◽  
Digital Image Analysis ◽  
Surface Coverage ◽  
Structural Parameters ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Foodborne Diseases ◽  
Average Thickness

ABSTRACT Because listeriosis is one of the deadliest foodborne diseases, controlling and eradicating Listeria monocytogenes biofilms is a serious challenge for food safety. Biofilms (24 h old) formed on polystyrene by a L. monocytogenes strain of food origin were exposed for a further 24 h to 12 different concentrations (from 100 to 1011 PFU/mL) of the bacteriophage P100 (Listex P100). The structural parameters of biofilms were studied by using confocal laser scanning microscopy and digital image analysis. The biovolume in the observation field (14,121 μm2) of control (untreated) biofilms was 237,333.1 ± 2,692.6 μm3. The biomass of treated biofilms ranged from 164.7 ± 89.0 μm3 (biofilms exposed to 1010 PFU/mL) to 231,170.5 ± 15,142.0 μm3 (100 PFU/mL). The lowest biomass was achieved after treatment with 108 PFU/mL, with no further decrease in biovolume when higher phage concentrations were used. A strong (P &lt; 0.001) correlation was found between phage concentration (log units) and biovolume (−0.965), surface coverage (−0.939), roughness (0.976), maximum thickness (−0.853), and average thickness (−0.965). Findings from this research suggest that bacteriophage P100 at concentrations equal to or greater than 8 log PFU/mL successfully removes L. monocytogenes biofilms from polystyrene surfaces.

scholarly journals High Rates of Conjugation in Bacterial Biofilms as Determined by Quantitative In Situ Analysis

1999 ◽  
Vol 65 (8) ◽  
pp. 3710-3713 ◽  
Author(s):  
Martina Hausner ◽  
Stefan Wuertz
Keyword(s):  
Laser Scanning ◽  
Nutrient Concentration ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Bacterial Biofilms ◽  
Conjugative Gene Transfer ◽  
Biofilm Structure

ABSTRACT Quantitative in situ determination of conjugative gene transfer in defined bacterial biofilms using automated confocal laser scanning microscopy followed by three-dimensional analysis of cellular biovolumes revealed conjugation rates 1,000-fold higher than those determined by classical plating techniques. Conjugation events were not affected by nutrient concentration alone but were influenced by time and biofilm structure.

scholarly journals Determination of the pH Gradient in Hair Follicles of Human Volunteers Using pH-Sensitive Melamine Formaldehyde-Pyranine Nile Blue Microparticles

Sensors ◽  
2020 ◽  
Vol 20 (18) ◽  
pp. 5243 ◽  
Author(s):  
Dennis Kaden ◽  
Lars Dähne ◽  
Fanny Knorr ◽  
Heike Richter ◽  
Jürgen Lademann ◽  
...  
Keyword(s):  
Hair Follicle ◽  
Laser Scanning ◽  
Nile Blue ◽  
Hair Shaft ◽  
Hair Follicles ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Melamine Formaldehyde

Nanoparticles can be applied to the hair follicles, which can serve as reservoirs for triggered drug release. A valid measurement method for the determination of the pH within the hair follicle in vivo has not been shown yet. Here, melamine formaldehyde particles up to 9 µm in size were applied on 40 freshly plucked scalp hairs of eight individuals to determine the pH along the hair shaft down to the root area of the hair. For fluorescent pH indicators, pyranine and Nile blue were incorporated into the particles. Measurements were conducted using confocal laser scanning microscopy. A pH decay gradient could be found from the hair sheath towards the external hair shaft (p = 0.012) with pH values at the hair sheath of 6.63 ± 0.09, at the hair sheath end at 6.33 ± 0.11, and at the external hair shaft at 6.17 ± 0.09 (mean ± SE). The pH difference between the hair sheath end and the external hair shaft was found to be significant (p = 0.036). The results might be comparable with the pH within the hair follicle in vivo indicating a pH increase towards the hair root.

scholarly journals Genetic Features of Resident Biofilms Determine Attachment of Listeria monocytogenes

2009 ◽  
Vol 75 (24) ◽  
pp. 7814-7821 ◽  
Author(s):  
Olivier Habimana ◽  
Mickael Meyrand ◽  
Thierry Meylheuc ◽  
Saulius Kulakauskas ◽  
Romain Briandet
Keyword(s):  
Listeria Monocytogenes ◽  
Laser Scanning ◽  
Time Course ◽  
Surface Hydrophobicity ◽  
Flow Cell ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Microscopy Imaging ◽  
Initial Fixation ◽  
Genetic Features

ABSTRACT Planktonic Listeria monocytogenes cells in food-processing environments tend most frequently to adhere to solid surfaces. Under these conditions, they are likely to encounter resident biofilms rather than a raw solid surface. Although metabolic interactions between L. monocytogenes and resident microflora have been widely studied, little is known about the biofilm properties that influence the initial fixation of L. monocytogenes to the biofilm interface. To study these properties, we created a set of model resident Lactococcus lactis biofilms with various architectures, types of matrices, and individual cell surface properties. This was achieved using cell wall mutants that affect bacterial chain formation, exopolysaccharide (EPS) synthesis and surface hydrophobicity. The dynamics of the formation of these biofilm structures were analyzed in flow cell chambers using in situ time course confocal laser scanning microscopy imaging. All the L. lactis biofilms tested reduced the initial immobilization of L. monocytogenes compared to the glass substratum of the flow cell. Significant differences were seen in L. monocytogenes settlement as a function of the genetic background of resident lactococcal biofilm cells. In particular, biofilms of the L. lactis chain-forming mutant resulted in a marked increase in L. monocytogenes settlement, while biofilms of the EPS-secreting mutant efficiently prevented pathogen fixation. These results offer new insights into the role of resident biofilms in governing the settlement of pathogens on food chain surfaces and could be of relevance in the field of food safety controls.

scholarly journals Characterization of Binary Biofilms of Listeria monocytogenes and Lactobacillus and Their Response to Chlorine Treatment

2021 ◽  
Vol 12 ◽  
Author(s):  
Magdalena A. Olszewska ◽  
Francisco Diez-Gonzalez
Keyword(s):  
Listeria Monocytogenes ◽  
Laser Scanning ◽  
Spatial Organization ◽  
Volume Ratio ◽  
Single Species ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Interspecies Interactions ◽  
Biofilm Thickness ◽  
Cell Counts

In nature, Listeria may interact competitively and cooperatively with other organisms, resulting in unique spatial organization and functions for cells within the community. This study was undertaken to characterize the biofilm architecture of binary biofilms of Listeria monocytogenes and Lactobacillus species and to assess their effect on the survival of Listeria during exposure to hypochlorite. Three L. monocytogenes strains, ATCC 19115 (Lm5), ATCC 19117 (Lm7), and Coleslaw (LmC), were selected and combined individually with three Lactobacillus strains: L. fermentum (Lf), L. bavaricus (Lb), and L. plantarum (Lp). In binary Lm-Lp biofilms, the Lm cell counts were similar to single-species biofilms (8.5 log CFU/well), and the Lp cell numbers declined by 1.0 log CFU/well. In the presence of Lb, the Lm cell counts were reduced by 1.5 log CFU/well (p &lt; 0.05), whereas the Lf cell counts increased at least by 3.5 log CFU/well. Confocal laser scanning microscopy (CLSM) determined that interspecies interactions significantly affected the spatial organization of three binary biofilms. Biofilm surface-to-volume ratio increased from 0.8 μm2/μm3 for Lm5 in the monoculture to 2.1 μm2/μm3 for Lm5-Lp in the dual-species model (p &lt; 0.05), and was characterized by a thicker structure with a largely increased surface area. Biofilm roughness increased from 0.2 for Lm7 to 1.0 for Lm7-Lb biofilms (p &lt; 0.05), which appeared as interspecific segregation. Biofilm thickness increased from 34.2 μm for LmC to 46.3 μm for LmC–Lf (p &lt; 0.05), which produced flat and compact structures that covered the entire surface available. The biomass of the extracellular matrix was higher in the case of some binary biofilms (p &lt; 0.05); however, this effect was dependent upon the species pair. When treated with hypochlorite, Lm5 in binary biofilms had an approximately 1.5 log CFU/well greater survival than individually. The unique spatial organization and greater protein production may explain the protective effect of Lp after hypochlorite exposure.

A Simple and Rapid Staining Technique for Sex Determination of Trichinella Larvae Parasites by Confocal Laser Scanning Microscopy

2019 ◽  
Vol 25 (6) ◽  
pp. 1491-1497 ◽  
Author(s):  
Inese Gavarane ◽  
Elena Kirilova ◽  
Ilze Rubeniņa ◽  
Ligita Mežaraupe ◽  
Sergejs Osipovs ◽  
...  
Keyword(s):  
Sex Determination ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Staining Technique ◽  
Life Cycle Stage ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser

AbstractThe roundworms of Trichinella genus are worldwide distributed and their prevalence in nature is high. Trichinella genus parasites are the causative agents of foodborne zoonosis trichinellosis. The main prevention and control of the infection are meat inspection by the magnetic stirrer method for the detection of Trichinella larvae in muscle samples. The treatment can be effective if the parasite is discovered early in the intestinal phase. Once the Trichinella larva has reached the muscle tissue, the parasite remains therein and there is no treatment for this life cycle stage. The Trichinella species is dioecious with separate male and female individuals. The developed staining technique that uses confocal laser scanning microscopy (CLSM) displays sufficient results for Trichinella larvae examination and this protocol is applicable to study the internal and external structures and for the sex determination of T. britovi and T. spiralis larvae samples. In the present study, a luminescent derivative was synthesized and used for staining of T. spiralis and T. britovi larvae samples for the examination by CLSM. Various fixatives, such as AFA, 70% ethanol, and Bouin's and Carnoy's solutions were tested for sample preparation. The synthesized luminescent compound demonstrates best visualization results for samples fixed in Bouin's fixative.

scholarly journals Distribution of extracellular DNA in Listeria monocytogenes biofilm

2019 ◽  
Vol 37 (No. 6) ◽  
pp. 409-416
Author(s):  
Martina Šuláková ◽  
Jarmila Pazlarová ◽  
Rikke Louise Meyer ◽  
Kateřina Demnerová
Keyword(s):  
Listeria Monocytogenes ◽  
Laser Scanning ◽  
3D Structure ◽  
Laser Scanning Microscopy ◽  
Extracellular Dna ◽  
Confocal Laser ◽  
Matrix Component ◽  
Signal Distribution ◽  
Culturing Conditions ◽  
Phylogenetic Lineages

Extracellular DNA (eDNA) is an abundant matrix component that protects biofilm from environmental stress, facilitate horizontal gene transfer, and serve as a source of nutrients. eDNA is also found in Listeria monocytogenes biofilm, but it is unknown to which extent its importance as a matrix component varies in terms of phylogenetic relatedness. This study aims to determine if these variations exist. Biofilm forming capacity of ten L. monocytogenes strains of different phylogenetic lineages and serotypes was examined using crystal violet assay at 37°C and 22°C. eDNA content was evaluated fluorometrically at 37°C and at 22°C, then the 3D structure of biofilm was studied by confocal laser scanning microscopy (CLSM). Biofilm forming capacity differed significantly between the culturing conditions and was higher at 37°C than at ambient temperature. eDNA signal distribution was found to be influenced by strain and lineage. CLSM images revealed information about spatial distribution in the biofilm. The information about the eDNA spatial organisation in the biofilm contributes to the understanding of the role of eDNA in a biofilm formation.

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