scholarly journals Characterization of Biofilm Formation by Clinically Relevant Serotypes of Group A Streptococci

2006 ◽  
Vol 72 (4) ◽  
pp. 2864-2875 ◽  
Author(s):  
Cordula Lembke ◽  
Andreas Podbielski ◽  
Carlos Hidalgo-Grass ◽  
Ludwig Jonas ◽  
Emanuel Hanski ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Laser Scanning ◽  
Group A Streptococcus ◽  
Three Dimensional ◽  
Positive Control ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Subsequent Formation ◽  
Group A

ABSTRACT Streptococcus pyogenes (group A streptococcus [GAS]) is a frequent cause of purulent infections in humans. As potentially important aspects of its pathogenicity, GAS was recently shown to aggregate, form intratissue microcolonies, and potentially participate in multispecies biofilms. In this study, we show that GAS in fact forms monospecies biofilms in vitro, and we analyze the basic parameters of S. pyogenes in vitro biofilm formation, using Streptococcus epidermidis as a biofilm-positive control. Of nine clinically important serotype strains, M2, M6, M14, and M18 were found to significantly adhere to coated and uncoated polystyrene surfaces. Fibronectin and collagen types I and IV best supported primary adherence of serotype M2 and M18 strains, respectively, whereas serotype M6 and M14 strains strongly bound to uncoated polystyrene surfaces. Absorption measurements of safranin staining, as well as electron scanning and confocal laser scanning microscopy, documented that primary adherence led to subsequent formation of three-dimensional biofilm structures consisting of up to 46 bacterial layers. Of note, GAS isolates belonging to the same serotype were found to be very heterogeneous in their biofilm-forming behavior. Biofilm formation was equally efficient under static and continuous flow conditions and consisted of the classical three steps, including partial disintegration after long-term incubation. Activity of the SilC signaling peptide as a component of a putative quorum-sensing system was found to influence the biofilm structure and density of serotype M14 and M18 strains. Based on the presented methods and results, standardized analyses of GAS biofilms and their impact on GAS pathogenicity are now feasible.

scholarly journals Bacterial lipopolysaccharides variably modulate in vitro biofilm formation of Candida species

2010 ◽  
Vol 59 (10) ◽  
pp. 1225-1234 ◽  
Author(s):  
H. M. H. N. Bandara ◽  
O. L. T. Lam ◽  
R. M. Watt ◽  
L. J. Jin ◽  
L. P. Samaranayake
Keyword(s):  
Biofilm Formation ◽  
Laser Scanning ◽  
Significant Inhibition ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Dependent Manner ◽  
Bacterial Lipopolysaccharides ◽  
Species Specific ◽  
Biofilm Development

The objective of this study was to evaluate the effect of the bacterial endotoxin LPS on Candida biofilm formation in vitro. The effect of the LPS of Pseudomonas aeruginosa, Klebsiella pneumoniae, Serratia marcescens and Salmonella typhimurium on six different species of Candida, comprising Candida albicans ATCC 90028, Candida glabrata ATCC 90030, Candida krusei ATCC 6258, Candida tropicalis ATCC 13803, Candida parapsilosis ATCC 22019 and Candida dubliniensis MYA 646, was studied using a standard biofilm assay. The metabolic activity of in vitro Candida biofilms treated with LPS at 90 min, 24 h and 48 h was quantified by XTT reduction assay. Viable biofilm-forming cells were qualitatively analysed using confocal laser scanning microscopy (CLSM), while scanning electron microscopy (SEM) was employed to visualize the biofilm structure. Initially, adhesion of C. albicans was significantly stimulated by Pseudomonas and Klebsiella LPS. A significant inhibition of Candida adhesion was noted for the following combinations: C. glabrata with Pseudomonas LPS, C. tropicalis with Serratia LPS, and C. glabrata, C. parapsilosis or C. dubliniensis with Salmonella LPS (P<0.05). After 24 h of incubation, a significant stimulation of initial colonization was noted for the following combinations: C. albicans/C. glabrata with Klebsiella LPS, C. glabrata/C. tropicalis/C. krusei with Salmonella LPS. In contrast, a significant inhibition of biofilm formation was observed in C. glabrata/C. dubliniensis/C. krusei with Pseudomonas LPS, C. krusei with Serratia LPS, C. dubliniensis with Klebsiella LPS and C. parapsilosis/C. dubliniensis /C. krusei with Salmonella LPS (P<0.05). On further incubation for 48 h, a significant enhancement of biofilm maturation was noted for the following combinations: C. glabrata/C. tropicalis with Serratia LPS, C. dubliniensis with Klebsiella LPS and C. glabrata with Salmonella LPS, and a significant retardation was noted for C. parapsilosis/C. dubliniensis/C. krusei with Pseudomonas LPS, C. tropicalis with Serratia LPS, C. glabrata/C. parapsilosis/C. dubliniensis with Klebsiella LPS and C. dubliniensis with Salmonella LPS (P<0.05). These findings were confirmed by SEM and CLSM analyses. In general, the inhibition of the biofilm development of LPS-treated Candida spp. was accompanied by a scanty architecture with a reduced numbers of cells compared with the profuse and densely colonized control biofilms. These data are indicative that bacterial LPSs modulate in vitro Candida biofilm formation in a species-specific and time-dependent manner. The clinical and the biological relevance of these findings have yet to be explored.

Endotracheal tubes coated with a broad-spectrum antibacterial ceragenin reduce bacterial biofilm in an in vitro bench top model

2021 ◽  
Author(s):  
María Consuelo Latorre ◽  
María Jesús Pérez-Granda ◽  
Paul B Savage ◽  
Beatriz Alonso ◽  
Pablo Martín-Rabadán ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Laser Scanning ◽  
In Vitro Study ◽  
Bacterial Biofilm ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Endotracheal Tubes ◽  
Clinical Strains ◽  
Number Of Cells

Abstract Background Ventilator-associated pneumonia is one of the most common nosocomial infections, caused mainly by bacterial/fungal biofilm. Therefore, it is necessary to develop preventive strategies to avoid biofilm formation based on new compounds. Objectives We performed an in vitro study to compare the efficacy of endotracheal tubes (ETTs) coated with the ceragenin CSA-131 and that of uncoated ETTs against the biofilm of clinical strains of Pseudomonas aeruginosa (PA), Escherichia coli (EC) and Staphylococcus aureus (SA). Methods We applied an in vitro bench top model using coated and uncoated ETTs that were treated with three different clinical strains of PA, EC and SA for 5 days. After exposure to biofilm, ETTs were analysed for cfu count by culture of sonicate and total number of cells by confocal laser scanning microscopy. Results The median (IQR) cfu/mL counts of PA, EC and SA in coated and uncoated ETTs were, respectively, as follows: 1.00 × 101 (0.0–3.3 × 102) versus 3.32 × 109 (6.6 × 108–3.8 × 109), P &lt; 0.001; 0.0 (0.0–5.4 × 103) versus 1.32 × 106 (2.3 × 103–5.0 × 107), P &lt; 0.001; and 8.1 × 105 (8.5 × 101–1.4 × 109) versus 2.7 × 108 (8.6 × 106–1.6 × 1011), P = 0.058. The median (IQR) total number of cells of PA, EC and SA in coated and non-coated ETTs were, respectively, as follows: 11.0 [5.5–not applicable (NA)] versus 87.9 (60.5–NA), P = 0.05; 9.1 (6.7–NA) versus 62.6 (42.0–NA), P = 0.05; and 97.7 (94.6–NA) versus 187.3 (43.9–NA), P = 0.827. Conclusions We demonstrated significantly reduced biofilm formation in coated ETTs. However, the difference for SA was not statistically significant. Future clinical studies are needed to support our findings.

scholarly journals Apicomplexan co-infections impair with phagocytic activity in avian macrophages

2020 ◽  
Vol 119 (12) ◽  
pp. 4159-4168
Author(s):  
Runhui Zhang ◽  
Wanpeng Zheng ◽  
Arwid Daugschies ◽  
Berit Bangoura
Keyword(s):  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Three Dimensional Model ◽  
Host Interactions ◽  
Macrophage Phagocytosis ◽  
High Doses ◽  
Free Ranging

AbstractMixed infections of Toxoplasma gondii and Eimeria tenella are likely to occur frequently due to the high prevalence of both pathogens in free-ranging chickens. In this study, we investigated the co-occurrence of the two parasites in the same immune-competent host cell towards altered patterns of parasite-host interactions. Chicken blood monocyte–derived macrophages were co-infected with T. gondii RH tachyzoites and E. tenella Houghton sporozoites in vitro for 24 h. Through monitoring the uptake of pH-sensitive pHrodo™ Zymosan BioParticles (“Zymosan”) by macrophages, we created a three-dimensional model and to analyze quantitatively phagocytosis using confocal laser scanning microscopy. Assessments of parasite populations were performed by qPCR at 2, 6, 12, and 24 h post-infection (hpi). At 6 hpi, phagocytosis was inhibited in the E. tenella–infected cultures while no inhibition of phagocytosis was observed due to T. gondii. Phagocytosis activity revealed more complex interactions during co-infection. At 12 and 24 hpi, phagocytosis response to “Zymosan” was distinctly weaker in co-infected cells than in all other groups except for cells mono-infected with high doses of E. tenella at 24 hpi. By qPCR, significantly reduced numbers of both intracellular parasites were recorded (10-fold) in all infected groups at 2 hpi. At 12 hpi, the T. gondii population reached lowest values but dramatically increased by 24 hpi. Our data confirm that macrophage phagocytosis is involved in the control of invasion by apicomplexan parasites in chicken which particularly applies to E. tenella infection and it was able to be altered by the co-existing parasites.

scholarly journals Peeking under the Iron Curtain: Development of a Microcosm for Imaging the Colonization of Steel Surfaces by Mariprofundus sp. Strain DIS-1, an Oxygen-Tolerant Fe-Oxidizing Bacterium

2016 ◽  
Vol 82 (22) ◽  
pp. 6799-6807 ◽  
Author(s):  
Adam C. Mumford ◽  
Irini J. Adaktylou ◽  
David Emerson
Keyword(s):  
Laser Scanning ◽  
Three Dimensional ◽  
Experimental System ◽  
Laser Scanning Microscopy ◽  
Dimensional Structure ◽  
Confocal Laser ◽  
Sterile Seawater ◽  
Microbially Influenced Corrosion ◽  
Steel Surfaces

ABSTRACTMicrobially influenced corrosion (MIC) is a major cause of damage to steel infrastructure in the marine environment. Despite their ability to grow directly on Fe(II) released from steel, comparatively little is known about the role played by neutrophilic iron-oxidizing bacteria (FeOB). Recent work has shown that FeOB grow readily on mild steel (1018 MS) incubatedin situor as a substrate for pure culturesin vitro; however, details of how they colonize steel surfaces are unknown yet are important for understanding their effects. In this study, we combine a novel continuously upwelling microcosm with confocal laser scanning microscopy (CLSM) to determine the degree of colonization of 1018 MS by the marine FeOB strain DIS-1. 1018 MS coupons were incubated with sterile seawater (pH 8) inoculated with strain DIS-1. Incubations were performed both under oxic conditions and in an anoxic-to-oxic gradient. Following incubations of 1 to 10 days, the slides were removed from the microcosms and stained to visualize both cells and stalk structures. Stained coupons were visualized by CLSM after being mounted in a custom frame to preserve the three-dimensional structure of the biofilm. The incubation of 1018 MS coupons with strain DIS-1 under oxic conditions resulted in initial attachment of cells within 2 days and nearly total coverage of the coupon with an ochre film within 5 days. CLSM imaging revealed a nonadherent biofilm composed primarily of the Fe-oxide stalks characteristic of strain DIS-1. When incubated with elevated concentrations of Fe(II), DIS-1 colonization of 1018 MS was inhibited.IMPORTANCEThese experiments describe the growth of a marine FeOB in a continuous culture system and represent direct visualizations of steel colonization by FeOB. We anticipate that these experiments will lay the groundwork for studying the mechanisms by which FeOB colonize steel and help to elucidate the role played by marine FeOB in MIC. These observations of the interaction between an FeOB, strain DIS-1, and steel suggest that this experimental system will provide a useful model for studying the interactions between microbes and solid substrates.

scholarly journals Antibody response to the 45 kDa Candida albicans antigen in an animal model and potential role of the antigen in adherence

2008 ◽  
Vol 57 (12) ◽  
pp. 1466-1472 ◽  
Author(s):  
Helena Bujdáková ◽  
Ema Paulovičová ◽  
Silvia Borecká-Melkusová ◽  
Juraj Gašperík ◽  
Soňa Kucharíková ◽  
...  
Keyword(s):  
Animal Model ◽  
Candida Albicans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Vaccine Development ◽  
Laser Scanning Microscopy ◽  
Model Systems ◽  
Confocal Laser

The Candida antigen CR3-RP (complement receptor 3-related protein) is supposed to be a ‘mimicry’ protein because of its ability to bind antibody directed against the α subunit of the mammalian CR3 (CD11b/CD18). This study aimed to (i) investigate the specific humoral isotypic response to immunization with CR3-RP in vivo in a rabbit animal model, and (ii) determine the role of CR3-RP in the adherence of Candida albicans in vitro using the model systems of buccal epithelial cells (BECs) and biofilm formation. The synthetic C. albicans peptide DINGGGATLPQ corresponding to 11 amino-acids of the CR3-RP sequence DINGGGATLPQALXQITGVIT, determined by N-terminal sequencing, was used for immunization of rabbits to obtain polyclonal anti-CR3-PR serum and for subsequent characterization of the humoral isotypic response of rabbits. A significant increase of IgG, IgA and IgM anti-CR3-RP specific antibodies was observed after the third (P<0.01) and the fourth (P<0.001) immunization doses. The elevation of IgA levels suggested peptide immunomodulation of the IgA1 subclass, presumably in coincidence with Candida epithelial adherence. Blocking CR3-RP with polyclonal anti-CR3-RP serum reduced the ability of Candida to adhere to BECs, in comparison with the control, by up to 35 % (P<0.001), and reduced biofilm formation by 28 % (P<0.001), including changes in biofilm thickness and integrity detected by confocal laser scanning microscopy. These properties of CR3-RP suggest that it has potential for future vaccine development.

scholarly journals Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract

2020 ◽  
Author(s):  
S. N. Sarah Shafiei ◽  
K. Ahmad ◽  
N. F. M. Ikhsan ◽  
S. I. Ismail ◽  
K. Sijam
Keyword(s):  
Biofilm Formation ◽  
Laser Scanning ◽  
Leaf Extract ◽  
Antimicrobial Agents ◽  
Acacia Mangium ◽  
Positive Control ◽  
Laser Scanning Microscopy ◽  
Xanthomonas Oryzae ◽  
Confocal Laser ◽  
Biofilm Inhibition

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

scholarly journals Influence of Remineralizing Dentifrice in the Treatment of Erosive Enamel Lesions

2018 ◽  
Vol 20 (4) ◽  
pp. 238
Author(s):  
Júlia Bazaga Ferreira ◽  
Gabriella Rodovalho Paiva ◽  
Vinícius Rangel Geraldo-Martins ◽  
Juliana Jendiroba Faraoni ◽  
Regina Guenka Palma Dibb ◽  
...  
Keyword(s):  
Surface Roughness ◽  
Laser Scanning ◽  
Tooth Enamel ◽  
In Vitro Study ◽  
Positive Control ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Significant Difference ◽  
Kolmogorov Smirnov

O objetivo deste trabalho in vitro foi avaliar a influência de diferentes agentes remineralizantes no tratamento de lesões erosivas em esmalte. Foram confeccionados espécimes de 4mmx4mm e 3 mm de espessura a partir da superfície vestibular de incisivos bovinos (n=10) e divididos aleatoriamente em 4 grupos. G1=aplicação do dentifrício remineralizante, G2= aplicação do agente potencializador remineralizante, G3= dentifrício remineralizante + agente potencializador remineralizante, G4=aplicação de verniz fluoretado (controle positivo), G5=nenhum tratamento (controle negativo). Os espécimes foram imersos em refrigerante durante um período de 10 dias. A rugosidade superficial foi analisada por meio de microscopia confocal de varredura a laser. Os dados foram analisados quanto à homogeneidade (Levene’s) e normalidade (Kolmogorov- Smirnov). Foram realizados testes paramétricos com análise de variância a dois critérios: fator tempo e fator tratamento, e pós-teste de Tukey para diferenciação das médias. Todos os testes estatísticos tiveram nível de significância de 5% (α=0,05). Os resultados obtidos mostraram diferenças estatisticamente significantes, demonstrando a redução da rugosidade da superfície do esmalte logo após o primeiro tratamento (G3) e para os demais grupos (G1, G2 e G4) somente após o segundo tratamento. Concluiu-se que a utilização de dentifrício composto por silicato de cálcio e fosfato de sódio influenciou na rugosidade do esmalte erodido do dente bovino.Palavras-chave: Dentifrícios. Erosão Dentária. Esmalte Dentário.Abstract The objective of this in vitro study was to evaluate the influence of different remineralizing agents in the treatment of enamel erosive lesions. Specimens of 4mmx4mm and 3mm thickness were made from the buccal surface of bovine incisors (n=10) and randomly divided into 4 groups. G1 = application of the remineralizing dentifrice, G2 = application of the remineralizing agent, G3 = remineralizing dentifrice + remineralizing agente, G4 = application of fluoride varnish (positive control), G5 = no treatment Specimens were immersed in refrigerant solution during a period of 10 days. The surface roughness was analyzed by means of confocal laser scanning microscopy. The data were analyzed for homogeneity (Levene's) and normality (Kolmogorov-Smirnov). Parametric tests with analysis of variance were performed on two criteria: time factor and treatment factor, and Tukey post-test for differentiation of means. All tests were statistically significant at 5% (α = 0.05). The results showed statistically significant difference, demonstrating the reduction of surface roughness after the first treatment (G3) and the other groups (G1, G2 and G4) only after the second treatment. It was concluded that the use of dentifrice composed of calcium silicate and sodium phosphate influenced the roughness of the eroded tooth enamel of the bovine tooth.Keywords: Dentifrices. Tooth Erosion. Tooth Enamel.

scholarly journals Biofilm Forming Ability Of Salmonella Enteritidis In Vitro

2015 ◽  
Vol 65 (3) ◽  
pp. 371-389 ◽  
Author(s):  
Čabarkapa Ivana ◽  
Škrinjar Marija ◽  
Lević Jovanka ◽  
Kokić Bojana ◽  
Blagojev Nevena ◽  
...  
Keyword(s):  
Crystal Violet ◽  
Congo Red ◽  
Laser Scanning ◽  
Salmonella Enteritidis ◽  
Incubation Temperature ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Microtiter Plates

AbstractSalmonella entericaserotype Enteritidis is an important alimentary pathogen that recently gained special attention due to the ability of a large number of strains to form biofilms. Qualitative testing of biofilm forming ability was performed by observing the morphotype of the colonies on Congo Red agar and by conducting the pellicle test, while quantitative testing was carried out by Cristal violet assay on microtiter plates. A total of 14 isolates of S. Enteritidis were tested for biofilm forming ability, while Salmonella Enteritidis ATTC 13076 was used as the reference strain. Based on the morphotype of colonies cultivated on Congo Red agar at 25°C incubation temperature, among tested isolates three morphotypes were detected – red, dry and rough (rdar), brown, dry and rough (bdar) and smooth and white (saw). Half of the tested isolates demonstrated rdar morphotype. All isolates that showed a specific morphotype at this incubation temperature also formed the corresponding type of pellicle at the air-liquid interface. Additionally, comparing OD (optical density) values obtained by crystal violet test between groups of isolates that represent one of the three detected morphotypes (rdar, bdar and saw), statistically significant differences were detected. Based on OD values obtained by crystal violet test at both applied incubation temperatures, isolates were classified into three categories, regarding their ability to form biofilms: strong, moderate and weak biofilm producers. By comparing the amounts of the biofilms formed after 48h at 25°C and 37°C, statistically significant differences were noted (P<0.05). In this research we presented micrographs and a reconstruction of three-dimensional projections of biofilm developing phases of rdar morphotype isolates, which were obtained using confocal laser scanning microscopy.

scholarly journals Spatial Arrangements and Associative Behavior of Species in an In Vitro Oral Biofilm Model

2001 ◽  
Vol 67 (3) ◽  
pp. 1343-1350 ◽  
Author(s):  
M. Guggenheim ◽  
S. Shapiro ◽  
R. Gmür ◽  
B. Guggenheim
Keyword(s):  
Biofilm Formation ◽  
Laser Scanning ◽  
Fusobacterium Nucleatum ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Streptococcus Sobrinus ◽  
Growth Phenomenon ◽  
Associative Behavior ◽  
Species Specific

ABSTRACT The spatial arrangements and associative behavior ofActinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis strains in an in vitro model of supragingival plaque were determined. Using species-specific fluorescence-labeled antibodies in conjunction with confocal laser scanning microscopy, the volumes and distribution of the five strains were assessed during biofilm formation. The volume-derived cell numbers of each strain correlated well with respective culture data. Between 15 min and 64 h, populations of each strain increased in a manner reminiscent of batch growth. The microcolony morphologies of all members of the consortium and their distributions within the biofilm were characterized, as were interspecies associations. Biofilms formed 15 min after inoculation consisted principally of single nonaggregated cells. All five strains adhered strongly to the saliva-conditioned substratum, and therefore, coadhesion played no role during the initial phase of biofilm formation. This observation does not reflect the results of in vitro coaggregation of the five strains, which depended upon the nature of the suspension medium. While the possibility cannot be excluded that some interspecies associations observed at later stages of biofilm formation were initiated by coadhesion, increase in bacterial numbers appeared to be largely a growth phenomenon regulated by the prevailing cultivation conditions.

scholarly journals Three-dimensional distribution of GFP-labeled Pseudomonas putida during biofilm formation on solid PAHs assessed by confocal laser scanning microscopy

2003 ◽  
Vol 47 (5) ◽  
pp. 139-142 ◽  
Author(s):  
A.C. Rodrigues ◽  
S. Wuertz ◽  
A.G. Brito ◽  
L.F. Melo
Keyword(s):  
Pseudomonas Putida ◽  
Confocal Laser Scanning Microscopy ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
In Situ Experiments

Confocal laser scanning microscopy was used to monitor the colonization pattern of the gfp-labeled derivative strain of Pseudomonas putida ATCC 17514 on fluorene and phenanthrene crystals. The in situ experiments showed that P. putida tends to grow directly on phenanthrene, forming a biofilm on accessible crystalline surfaces. On the other hand, no significant biofilm formation was observed in the presence of fluorene. The results obtained showed that substrate properties affected bacterial strategy regarding uptake.

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