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Concentrations of test microorganisms grown in various test media after centrifuging and resuspending in PBS
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Microorganisms have been shown to adhere to food-contact surfaces and may provide a route for the contamination of processed food. To better understand this phenomenon, the effects of growth media and surface conditioning on the adherence of Pseudomonas fragi, Salmonella typhimurium and Listeria monocytogenes cells to stainless steel were studied....
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Citations
... The presence of glucose in growth media has been reported to enhance biofilm formation (Stepanovi et al., 2000). Type of media, culture condition and surface of ves-sel cultivation are significantly robust the biofilm formation (Hood and Zottola, 1997;Iversen et al., 2004). The ability to adhere to a solid surface and the consecutive formation of an organised bacterial biofilm community are crucial for the formation of P. aeruginosa and S. pyogenes biofilms. ...
... Study by (Jaffar et al., 2016) reported that day 2 was the preferable incubation duration to produce biofilm in A. actinomycetemcomitans and day 7 for P. gingivalis. Previous studies have indicated that temperature, nutrients and other components in media affected the attachment of microorganisms to the surfaces of various materials (Hood and Zottola, 1997;Iversen et al., 2004). Study by (Al-kafaween et al., 2019b) showed that the E. coli produced biofilm on day 3 and day 5 in Brain heart infusion (BHI) and Luria broth (LB) at OD 0.05 and OD 0.1 (Al-kafaween et al., 2019). ...
Introduction: Biofilm formation has different stages and can be classified based on the bacterial strain, culture vessel, and the method employed. Biofilm formation is carried out in culture vessels to represent mode of infection in humans. Microbial concentration, growth medium, supplement, and incubation time are key factors to successfully form biofilm in a culture vessel. Objective: This study aimed to identify the optimum conditions for biofilm formation in a 96-well plate by culturing Pseudomonas aeruginosa and Streptococcus pyogenes. Materials and Methods: We utilized the infectious and pathogenic bacteria, P. aeruginosa and S. pyogenes strains. These bacteria were cultured in Mueller-Hinton Broth (MHB) and Tryptic Soy Broth (TSB) at two different optical densities (OD 0.05 and OD 0.1). After a certain incubation time, the formed biofilm was stained by using 0.1% crystal violet. The stained bacteria were disaggregated and measured using a microplate reader. Biofilm was then classified based on bacterial adherence to the plate. Results: Our results showed that P. aeruginosa and S. pyogenes biofilms were strongly formed on days 3 and 5 in MHB and TSB, respectively. However, the strongest biofilm formation was seen on day 3 after P. aeruginosa being incubated in MHB at OD 0.1 and after S. pyogenes being incubated in MHB at OD 0.05. Discussion: Biofilm formation is ranged between weak, moderate, and strong in accordance with the density of bacterial adhesion. Conclusion: P. aeruginosa and S. pyogenes biofilms were optimized at specific OD (0.1 and 0.05, respectively) for 3 days cultivation in MHB. Citation: Al-kafaween, M.A. and Hilmi A.B.M., 2022. Evaluation of the effect of different growth media and incubation time on the suitability of biofilm formation by Pseudomonas aeruginosa and Streptococcus pyogenes. Applied Environmental Biotechnology, 6(2):
... The EPS provides protection and trap nutrients for microorganisms in the biofilm, contributing to the formation of the cluster (Figure 4A). However, the adhesion of bacteria on surfaces is not related to the production of EPS, because adhesion and consequent biofilm formation can occur even in the absence of the polymeric substance (Hoodt and Zottola, 1997). Despite lack of information on the molecular mechanisms, such as genes and EPS characteristics involved in biofilm formation (Park-ji et al., 2019), there are reports in the literature that EPS production depends on stress and environmental conditions (Giaouris et al., 2015). ...
... The adhesion of B. cereus to surfaces also contributes to the formation of a conditioning layer, facilitating subsequent adhesion of other bacteria (Marchand et al., 2012), and the capability to form biofilms under static conditions (Wijman et al., 2007), because of their hydrophobic properties (Park-ji et al., 2019). Herald and Zottola (1988) and Hoodt and Zottola (1997) found that Listeria species and Yersinia species showed an increased adhesion capacity during high metabolic activity. Thus, it is also possible that in our experiments the microorganisms were in this phase at the fourth day of incubation, facilitating the visualization of the species on the stainless steel in Group 3 (Figures 3, 4). ...
Microorganisms in dairy industries can form monospecies, dual-species, or multispecies biofilms, showing cooperative or competitive behaviors, which might contribute to the reduction of efficiency of cleaning and sanitization processes and eventually turn into a potential source of contamination. This study proposes to evaluate the behavior of Listeria monocytogenes in monospecies biofilms, cocultured with Bacillus cereus. The isolates were of dairy origin, and the selection occurred after studies of competition among species. The biofilm formations on AISI 304 stainless steel at 25°C in a stationary culture were analyzed to observe the cooperative or competitive interactions among species, as well as the effect of pre-adhered cells. Biofilm formation assays were performed in four experiments: Experiment 1: in the presence of strains of antagonistic substance producer B. cereus (+); Experiment 2: extract of the antagonistic substance of B. cereus; Experiment 3: pre-adhered cells of B. cereus; and Experiment 4: pre-adhered cells of L. monocytogenes. Subsequently, cooperative behavior was observed by scanning electron microscopy. The L. monocytogenes monospecies biofilm counts of greater than 5 log colony-forming units (CFU)/cm² were also observed in dual-species biofilms in the presence of B. cereus (non-producers of antagonist substance), showing cooperative behavior between species. However, in the presence of antagonistic substance produced by B. cereus, the counts were lower, 1.39 and 1.70 log CFU/cm² (p > 0.05), indicating that the antagonistic substance contributes to competitive interactions. These data are relevant for the development of new studies to control L. monocytogenes in the dairy industry.
... Dans l'industrie laitière, le film conditionnant formé est fonction de la concentration de lait. Ainsi, Hood et Zottola ont montré que le substrat conditionné par le lait dilué possède une meilleure adhérence pour la fixation d'agents pathogènes comparée à celle du lait entier (Hood et Zottola, 1997 (Fletcher, 1976). De même, Meadows (1971) a également montré cet effet inhibiteur de l'albumine alors que la caséine et la gélatine favorisent le processus de fixation microbien (Meadows, 1971). ...
Dans de nombreux secteurs, l'adhésion microbienne sur les surfaces est la source de multiples impacts négatifs. Cette étape est considérée comme préliminaire au développement de biofilm et peut être influencée par la présence d'un film conditionnant engendré par l'adsorption des protéines sur la surface. Ainsi, les stratégies visant une intervention au moment de la phase initiale d'adhésion représentent une approche appropriée pour prévenir la bio-contamination des surfaces et nécessitent une compréhension à l'échelle moléculaire. Dans ce contexte, les matériaux nanocomposites à base de nanoparticules d'argents (AgNPs) et de silice (SiO2) apparaissent comme des outils pertinents. Ce travail de thèse porte sur l'utilisation de substrats nanocomposites possédant une monocouche d'AgNPs exposées à leurs surfaces ou enterrées dans une matrice de SiO2plasma à une distance contrôlée de quelques nanomètres de la surface afin d'explorer, d'une part l'adhésion de protéines modèles (Sérum Albumine Bovine, DsRed et Fibronectine) et leurs changements conformationnels et d'autre part, la cinétique de détachement de la levure Candida albicans dans les différentes conditions. Les AgNPs sont bien connues pour leurs activités antimicrobiennes et présentent de plus, des propriétés optiques permettant de détecter des signatures moléculaires à leurs proximités. Suite à l'application de la spectroscopie Raman exaltée de surface en utilisant les couches nanocomposites à base d'AgNPs, la détection de trois conformations de la DsRed (protéine fluorescente rouge) adsorbée et déshydratée sur les substrats plasmoniques a été possible. Les résultats obtenus montrent que les changements conformationnels des protéines avec une forte cohérence interne sont réversibles. En parallèle, nous avons évalué la dynamique d'organisation et le comportement de la SAB, de la Fn et de la DsRed en contact avec des couches minces de silice ou additivées d'AgNPs. Les mesures des angles de contact des gouttelettes de différentes concentrations protéiques ont montré une interaction hydrophile croissante avec la SiO2th thermique. L'hydrophobicité de surface est modifiée pour les substrats nanocomposites. L'épaisseur et les propriétés optiques des couches protéiques adsorbées ont été évaluées par ellipsométrie spectroscopique. En fonction de la concentration de protéines dans solution les résultats montrent l'évolution d'une monocouche protéique non continue et non dense vers une monocouche plus compacte et plus complexe pour des concentrations élevées.[...]
... Under such circumstances conditioning is one of the first occasions to occur via the adsorption of food particles to surface. Surface conditioning and growth media both were found to affect the attachment of bacterial cells to stainless steel [19]. Microbial efficacy of D-glycin plus imipenem, D-glycin plus ceftriaxone, imipenem and ceftriaxone alone separately against biofilm producing E. coli isolated from different sources was determined was demonstrated in tables 4 and 5. Synergetic effect of D-glycin with impinem in the inhibition of biofilm formation was more than the antibiotic alone. ...
ABSTRACT The ability of many bacteria to adhere to surfaces and form biofilm makes the treatment of infections so difficult and has major consequences in a variety of industries such as food industry. Biofilm generates a persistent cause of contamination, thus, in the present study; inhibition of biofilm formation efficacy of the combination of D-glycin with antibiotics was compared with the antibiotic alone against Escherichia coli biofilm formation. It was found that the synergetic action of antibiotics with D-glycin was effective on inhibition of biofilm formation. The isolates ability to form biofilm was assayed using the tissue culture plate and Congo-red agar methods and findings of this study were source related; since food origin isolates were more biofilm producers when the Congo-red agar used as compared with tissue culture plate method. Minimum inhibitory concentration (MIC) of D-glycin, imipenem and ceftriaxone were determined. The combinations of D-glycin with antibiotics inhibited bacterial biofilm formation more than the antibiotics imipenem and ceftriaxone separately. Susceptibility test to eight antibiotics: Imipenem, ceftriaxone, ampicillin, amoxicilin, cephalexin, clarithromycin, gentamycin and tetracycline; was performed on planktonic cells, and the cells exhibited different sensitivity patterns to them depending on their source.
... Under such circumstances conditioning is one of the first occasions to occur via the adsorption of food particles to surface. Surface conditioning and growth media both were found to affect the attachment of bacterial cells to stainless steel [19]. Microbial efficacy of D-glycin plus imipenem, D-glycin plus ceftriaxone, imipenem and ceftriaxone alone separately against biofilm producing E. coli isolated from different sources was determined was demonstrated in tables 4 and 5. Synergetic effect of D-glycin with impinem in the inhibition of biofilm formation was more than the antibiotic alone. ...
ABSTRACT The ability of many bacteria to adhere to surfaces and form biofilm makes the treatment of infections so difficult and has major consequences in a variety of industries such as food industry. Biofilm generates a persistent cause of contamination, thus, in the present study; inhibition of biofilm formation efficacy of the combination of D-glycin with antibiotics was compared with the antibiotic alone against Escherichia coli biofilm formation. It was found that the synergetic action of antibiotics with D-glycin was effective on inhibition of biofilm formation. The isolates ability to form biofilm was assayed using the tissue culture plate and Congo-red agar methods and findings of this study were source related; since food origin isolates were more biofilm producers when the Congo-red agar used as compared with tissue culture plate method. Minimum inhibitory concentration (MIC) of D-glycin, imipenem and ceftriaxone were determined. The combinations of D-glycin with antibiotics inhibited bacterial biofilm formation more than the antibiotics imipenem and ceftriaxone separately. Susceptibility test to eight antibiotics: Imipenem, ceftriaxone, ampicillin, amoxicilin, cephalexin, clarithromycin, gentamycin and tetracycline; was performed on planktonic cells, and the cells exhibited different sensitivity patterns to them depending on their source.
... In this study, L. monocytogenes and L. innocua formed a biofilm in an acidic environment with high sugar content at 25 C. Previously, it was reported that L. monocytogenes and L. innocua strains adhered to stainless steel at much lower levels than some of the other Gram positive and Gram negative isolates (Hood & Zottola 1997a, 1997bJeong & Frank 1994a, 1994b. In mono-species biofilms, L. innocua adhered on SSCs at significantly higher counts than L. monocytogenes. ...
When testing conditions that prevail in fruit juice industry it was found that Listeria monocytogenes ATCC 7644 and a wild strain of Listeria innocua were highly hydrophobic, expressed swimming and twitching, co‐aggregated with yeast cells and produced exopolysaccharide, all characteristics that would favor the adhesive process. In mono‐species biofilms, L. innocua adhered on stainless steel at significantly higher counts than L. monocytogenes achieving values of 6.64 ± 0.01 and 5.80 ± 0.21 log CFU/cm². In dual species biofilms with resident yeasts, L. innocua cells counts increase significantly in the presence of Candida tropicalis and Candida krusei and decrease significantly in the presence of Rhodotorula mucilaginosa and Candida kefyr. When cocultured with Candida tropicalis, the cell numbers of L. monocytogenes had a significant increase. These results revealed synergic and antagonistic interactions among species.
Practical applications
The study supports the plausibility that interactions between L. monocytogenes and members of resident microbiota, such as C. tropicalis, C. krusei, C. kefyr, and R. mucilaginosa, might play an important role for the survival and dissemination of L. monocytogenes. Apple juice processing conditions was used as simulation of the real condition in fruit juice processing environment and these results will alert to fruit juice industry to adopt the best cleaning and disinfecting practices against Listeria.
... Under such circumstances conditioning is one of the first occasions to occur via the adsorption of food particles to surface. Surface conditioning and growth media both were found to affect the attachment of bacterial cells to stainless steel [19]. Microbial efficacy of D-glycin plus imipenem, D-glycin plus ceftriaxone, imipenem and ceftriaxone alone separately against biofilm producing E. coli isolated from different sources was determined was demonstrated in tables 4 and 5. Synergetic effect of D-glycin with impinem in the inhibition of biofilm formation was more than the antibiotic alone. ...
The ability of many bacteria to adhere to surfaces and form biofilm makes the treatment of infections so difficult
and has major consequences in a variety of industries such as food industry. Biofilm generates a persistent cause of
contamination, thus, in the present study; inhibition of biofilm formation efficacy of the combination of D-glycin
with antibiotics was compared with the antibiotic alone against Escherichia coli biofilm formation. It was found that
the synergetic action of antibiotics with D-glycin was effective on inhibition of biofilm formation. The isolates ability
to form biofilm was assayed using the tissue culture plate and Congo-red agar methods and findings of this study
were source related; since food origin isolates were more biofilm producers when the Congo-red agar used as
compared with tissue culture plate method. Minimum inhibitory concentration (MIC) of D-glycin, imipenem and
ceftriaxone were determined. The combinations of D-glycin with antibiotics inhibited bacterial biofilm formation
more than the antibiotics imipenem and ceftriaxone separately. Susceptibility test to eight antibiotics: Imipenem,
ceftriaxone, ampicillin, amoxicilin, cephalexin, clarithromycin, gentamycin and tetracycline; was performed on
planktonic cells, and the cells exhibited different sensitivity patterns to them depending on their source.
... Em cada Erlenmeyer foram adicionados três cupons de aço inoxidável AISI 304 # 4 com dimensões de 2 cm x 2 cm x 0,1 cm, previamente higienizados e esterilizados de acordo com Rossoni & Gaylarde (2000). A superfície de aço inoxidável AISI 304 # 4 foi escolhida por ser a mais empregada em tanques de expansão de leite (Hood & Zottola, 1997). ...
The Myrtaceae family has
great occurrence in Brazil, mainly in the Atlantic Forest of the South and Southeast regions
and presents potential for essential oil production. This work aimed to evaluate the essential
oil yield and composition of the species Myrciaria delicatula, Campomanesia xantocarpha,
Campomanesia aurea, Calyptranthes clusiifolia, Myrcia splendens, Eugenia osoriana, Myrciaria
tenella, Myrceugenia reitzii, Calyptranthes concinna e Myrcia arborensis from a segment of the
Atlantic Forest of Parana State. The sample collections for essential oil extraction, photographic
records and herbarium specimens preparation for botanical species identification were made
at the Private Reserve of Natural Heritage (PRNP) Butuguara in Palmeira (PR), with “Campos
Gerais” formations. The identification and the official deposit of the species were made at the
HFIE Herbarium. The essential oil extraction was carried out by hydrodistillation of fresh and dried
leaves using a Clevenger type apparatus and the chemical composition was analyzed by gas
phase chromatography coupled with mass spectrometry (GC/MS). Myrceugenia reitzii showed
eesential oil yield superior to other species and the essential oil yield of all species increased
after drying. The chemical composition in the samples showed mono and sesquiterpenes,
being the sesquiterpenes in higher percentages. The drying conditions affected the essential
oil composition of the for the majority of the evaluated species.
... However, before a cell can bind to a surface, the surface is conditioned by adsorbing molecules from the surrounding environment. In brief, bacteria along with other organic and inorganic molecules gets adsorbed to the surface and, then, the accumulation of molecules at the solid-liquid interface on food-contact surfaces leads to a higher concentration of nutrients compared to the fluid phase and acts as a conditioning film (Hood and Zottola, 1997). This conditioning film favours biofilm formation and alters the physico-chemical properties of the surface, e.g., changes in hydrophobicity and electrostatic charges (Dickson and Koohmaraie, 1989). ...
... Studies have shown the capacity of several food-borne pathogens, such as L. monocytogenes, to persist in the environment for years Lunden et al. 2002). In particular, environmental surfaces such as floors and walls may be indirect sources of contamination e.g., transference to the food products by vectors such as air, people and cleaning systems (Holah, 1992;Gibson et al. 1999). In food systems, improperly cleaned and sanitized equipment and air-borne microflora are usually considered to be the major sources of contamination . ...
Listeria monocytogenes is a facultative intracellular bacterium and hence, this bacterium is able to survive and propagate in macrophages (M. Yet, in contrast to this notion, a majority of L. monocytogenes is killed by M indicating that M play a critical role in protection against L. monocytogenes infection. Because the number of L. monocytogenes engulfed and killed by M depends on whether or not M are activated, experimental listeriosis has been employed for analyzing M activities by many investigators. There are several methods for analyzing phagocytic and listericidal activities of M Yet, in most cases expensive materials and complicated methods are required. In this review, in addition to methods recently employed for determination of phagocytic and listericidal activities of M, simple and excellent, though old-fashioned, method is also introduced.
... However, before a cell can bind to a surface, the surface is conditioned by adsorbing molecules from the surrounding environment. In brief, bacteria along with other organic and inorganic molecules gets adsorbed to the surface and, then, the accumulation of molecules at the solid-liquid interface on food-contact surfaces leads to a higher concentration of nutrients compared to the fluid phase and acts as a conditioning film (Hood and Zottola, 1997). This conditioning film favours biofilm formation and alters the physico-chemical properties of the surface, e.g. ...
... In particular, environmental surfaces such as floors and walls may be indirect sources of contamination e.g. transference to the food products by vectors such as air, people and cleaning systems (Holah, 1992;Gibson et al. 1999). In food systems, improperly cleaned and sanitized equipment and air-borne microflora are usually considered to be the major sources of contamination (Simoes et al. 2010). ...
... L monocytogenes can adhere rapidly and firmly to inert surfaces commonly found in the meat processing industry (Autio et al. 1999;Lyytikainen et al. 2000;Miettinen et al. 2001;D'Orio et a., 2007). L. monocytogenes, in fact, is capable of adhering and forming biofilm on common materials used in meat industry such as metal, glass, rubber and plastic surfaces (Hood and Zottola, 1997;Chae andSchraft, 2000, 2006;Sinde and Cerballo, 2000;Joseph et al. 2001;Stepanovic et al. 2004;Moltz and Martin, 2005). ...
Listeria monocytogenes is an important foodborne pathogen not because it causes large numbers of symptomatic cases but because of its relatively high case–fatality rate (20-30%). Listeriosis appears to be primarily a foodborne infection and is particularly a problem on foods that are not cooked, including ready-to-eat (RTE) meats, soft cheeses, and unpasteurized dairy products, as well as vegetables. Thermal processing of milk and meat products will destroy L. monocytogenes but post-processing contamination does occur. L. monocytogenes that contaminates foods may originate in soils from farms, in slaughtering facilities, in food processing plants and other equipments used in food manufacture. Elimination of Listeria from many environmental sources can be challenging because of the resistant, persistent biofilms formed by this microorganism. Bacterial biofilms are generally described as surface-associated bacterial communities forming microcolonies surrounded by a matrix of exopolymers. Biofilm formation can be separated into four sequential steps: (i) conditioning of the surface, (ii) initial adherence, (iii) physical irreversible adherence that involves the production of exopolymers that fix the cells, and (iv) growth of the microorganisms, which form microcolonies, and coaggregation leading to establishment of the biofilm. Such structures are dynamic systems in which cells grow, die, and/or are released, allowing contamination of other surfaces. Moreover, biofilms confer to L. monocytogenes a greater resistance to antimicrobial agents and they allow microorganisms to resist to environmental changes and cleaning/disinfection treatments. In the following, through an up-to-date review of (personal and non) published data, the growth of L. monocytogenes in biofilm in the food industry will be discussed. ***Full text available upon request
