Phenotypes of Non-Attached Pseudomonas aeruginosa Aggregates Resemble Surface Attached Biofilm
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{"title"=>"Phenotypes of non-attached pseudomonas aeruginosa aggregates resemble surface attached biofilm", "type"=>"journal", "authors"=>[{"first_name"=>"Morten", "last_name"=>"Alhede", "scopus_author_id"=>"15755128600"}, {"first_name"=>"Kasper Nørskov", "last_name"=>"Kragh", "scopus_author_id"=>"54396954400"}, {"first_name"=>"Klaus", "last_name"=>"Qvortrup", "scopus_author_id"=>"55210215600"}, {"first_name"=>"Marie", "last_name"=>"Allesen-Holm", "scopus_author_id"=>"8137621000"}, {"first_name"=>"Maria", "last_name"=>"van Gennip", "scopus_author_id"=>"26029570900"}, {"first_name"=>"Louise D.", "last_name"=>"Christensen", "scopus_author_id"=>"17134466300"}, {"first_name"=>"Peter Østrup", "last_name"=>"Jensen", "scopus_author_id"=>"14525265100"}, {"first_name"=>"Anne K.", "last_name"=>"Nielsen", "scopus_author_id"=>"18634822100"}, {"first_name"=>"Matt", "last_name"=>"Parsek", "scopus_author_id"=>"7003840250"}, {"first_name"=>"Dan", "last_name"=>"Wozniak", "scopus_author_id"=>"7005575242"}, {"first_name"=>"Søren", "last_name"=>"Molin", "scopus_author_id"=>"57197278680"}, {"first_name"=>"Tim", "last_name"=>"Tolker-Nielsen", "scopus_author_id"=>"6603859418"}, {"first_name"=>"Niels", "last_name"=>"Høiby", "scopus_author_id"=>"35478208900"}, {"first_name"=>"Michael", "last_name"=>"Givskov", "scopus_author_id"=>"19334462000"}, {"first_name"=>"Thomas", "last_name"=>"Bjarnsholt", "scopus_author_id"=>"8570536600"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-81455127357", "pui"=>"362955807", "doi"=>"10.1371/journal.pone.0027943", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"81455127357", "pmid"=>"22132176"}, "id"=>"78263f6c-6698-3e10-b241-52804f404777", "abstract"=>"For a chronic infection to be established, bacteria must be able to cope with hostile conditions such as low iron levels, oxidative stress, and clearance by the host defense, as well as antibiotic treatment. It is generally accepted that biofilm formation facilitates tolerance to these adverse conditions. However, microscopic investigations of samples isolated from sites of chronic infections seem to suggest that some bacteria do not need to be attached to surfaces in order to establish chronic infections. In this study we employed scanning electron microscopy, confocal laser scanning microscopy, RT-PCR as well as traditional culturing techniques to study the properties of Pseudomonas aeruginosa aggregates. We found that non-attached aggregates from stationary-phase cultures have comparable growth rates to surface attached biofilms. The growth rate estimations indicated that, independently of age, both aggregates and flow-cell biofilm had the same slow growth rate as a stationary phase shaking cultures. Internal structures of the aggregates matrix components and their capacity to survive otherwise lethal treatments with antibiotics (referred to as tolerance) and resistance to phagocytes were also found to be strikingly similar to flow-cell biofilms. Our data indicate that the tolerance of both biofilms and non-attached aggregates towards antibiotics is reversible by physical disruption. We provide evidence that the antibiotic tolerance is likely to be dependent on both the physiological states of the aggregates and particular matrix components. Bacterial surface-attachment and subsequent biofilm formation are considered hallmarks of the capacity of microbes to cause persistent infections. We have observed non-attached aggregates in the lungs of cystic fibrosis patients; otitis media; soft tissue fillers and non-healing wounds, and we propose that aggregated cells exhibit enhanced survival in the hostile host environment, compared with non-aggregated bacterial populations.", "link"=>"http://www.mendeley.com/research/phenotypes-nonattached-pseudomonas-aeruginosa-aggregates-resemble-surface-attached-biofilm", "reader_count"=>146, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>7, "Researcher"=>21, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>36, "Student > Postgraduate"=>7, "Other"=>5, "Student > Master"=>32, "Student > Bachelor"=>26, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>7, "Researcher"=>21, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>36, "Student > Postgraduate"=>7, "Other"=>5, "Student > Master"=>32, "Student > Bachelor"=>26, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Engineering"=>8, "Unspecified"=>1, "Environmental Science"=>4, "Biochemistry, Genetics and Molecular Biology"=>14, "Nursing and Health Professions"=>1, "Medicine and Dentistry"=>18, "Agricultural and Biological Sciences"=>77, "Pharmacology, Toxicology and Pharmaceutical Science"=>5, "Business, Management and Accounting"=>1, "Physics and Astronomy"=>2, "Chemistry"=>5, "Immunology and Microbiology"=>10}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>8}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>18}, "Chemistry"=>{"Chemistry"=>5}, "Physics and Astronomy"=>{"Physics and Astronomy"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>10}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>77}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>14}, "Unspecified"=>{"Unspecified"=>1}, "Environmental Science"=>{"Environmental Science"=>4}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>5}}, "reader_count_by_country"=>{"Republic of Singapore"=>1, "United States"=>3, "Denmark"=>1, "Brazil"=>1, "United Kingdom"=>2, "France"=>1, "Switzerland"=>1, "Germany"=>2}, "group_count"=>5}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/710581"], "description"=>"<p>SEM of A – Aggregate harvested from a 48-h old stationary culture. B – Details of a 3-day old biofilm grown in flow-cell. C – Details of 48-h old stationary aggregate. D – Planktonic cells (OD<sub>600</sub> = 0.5). Note that the single planktonic cells are difficult to fixate on the specimen during SEM preparation due to the small size (leads to few cells on the specimen).</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>380905, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SEM_survey_of_different_P_aeruginosa_cultures_/380905", "title"=>"SEM survey of different <i>P. aeruginosa</i> cultures.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:29:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/710720"], "description"=>"<p>Mixed Yellow (YFP) and Cyan (CFP) constitually tagged PAO1 were inoculated together. The CLSM visualization demonstrated that the clumps consisted of both yellow and blue cells indicating non-clonal growth. Length of size bar: 6 µm.</p>", "links"=>[], "tags"=>["clonal"], "article_id"=>381065, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Invstigation_of_the_clonal_relationship_in_clumps_/381065", "title"=>"Invstigation of the clonal relationship in clumps.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:30:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/710845"], "description"=>"<p>Biofilms and aggregates were grown for 24 h to 72-h prior to tobramycin (100 ug/ml) treatment for 24-h. For visualization by CLSM a GFP-tagged PAO1 strain (green) was used and stained with the DNA stain PI (red) for visualizing dead bacteria. Panels A, B and C represent biofilm tolerance on day 1, 2 and 3 respectively andpanels D and E are aggregates on day 1 and 2. Length of size bars: 20 µm.</p>", "links"=>[], "tags"=>["maturing", "flow-cell", "biofilms", "aggregates", "harvested", "static"], "article_id"=>381184, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibiotic_tolerance_of_maturing_flow_cell_biofilms_of_P_aeruginosa_and_aggregates_harvested_from_a_static_P_aeruginosa_culture_/381184", "title"=>"Antibiotic tolerance of maturing flow-cell biofilms of <i>P. aeruginosa</i> and aggregates harvested from a static <i>P. aeruginosa</i> culture.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:30:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/710969"], "description"=>"<p>Growth rates were estimated by quantifying rRNA molecules per rDNA molecules by RT-PCR.</p>", "links"=>[], "tags"=>["rates", "flow-cell", "biofilms", "aggregates"], "article_id"=>381315, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Growth_rates_of_flow_cell_biofilms_and_aggregates_over_time_/381315", "title"=>"Growth rates of flow-cell biofilms and aggregates over time.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:31:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/711070"], "description"=>"<p>Aggregates were grown for 48-h prior to tobramycin (100 ug/ml) treatment for 24-h or whirly mix (20 seconds) followed by tobramycin treatment for 24 h. 24 h. Bars represent the median. Mann-Whitney test were chosen to compare the medians of the different treatment regimens. ns indicates no significant differences between treatments (p>0.05). * Indicates significant difference (p<0.05). *** Indicates a very significant difference (p<0.0001).</p>", "links"=>[], "tags"=>["48-h", "aggregates", "harvested", "static", "disruption", "whirly"], "article_id"=>381402, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibiotic_tolerance_of_48_h_old_aggregates_harvested_from_a_static_P_aeruginosa_culture_and_the_effect_of_disruption_by_whirly_mixing_/381402", "title"=>"Antibiotic tolerance of 48-h old aggregates harvested from a static <i>P. aeruginosa</i> culture and the effect of disruption by whirly mixing.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:31:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/711166"], "description"=>"<p>Aggregates were grown for 48-h prior to tobramycin (100 ug/ml) treatment for 24-h, followed by whirly mix and another round of tobramycin treatment. Number indicates seconds of whirly mix. Bars represent the median. Kruskal-Wallis with Dunn's post test was chosen to compare the effect of whirly mix and subsequent treatment with tobramycin. The post test found the increased killing to be significant (P<0.05 when compared to 0 seconds) after 30 seconds of whirly mix.</p>", "links"=>[], "tags"=>["prolonged", "whirly"], "article_id"=>381513, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_prolonged_whirly_mix_/381513", "title"=>"Effect of prolonged whirly mix.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:32:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/711255"], "description"=>"<p>Flow-cell biofilms (A+B) were grown for 72-h before addition of PMNs and the aggregates (C+D) were grown for 48-h before the addition of PMNs. The images shows that aggregates are not phagocytosed or penetrated by PMNs. For visualization a GFP-tagged PAO1 strain (green) was used and SYTO62 was used to stain the PMNs (red). Arrows point at paralyzed PMNs. Length of size bars: 20 µm.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>381601, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Tolerance_towards_PMNs_/381601", "title"=>"Tolerance towards PMNs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:32:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/711399"], "description"=>"<p>DNA (PI) staining of A - Aggregate harvested from a 48-h old stationary culture stained with PI. B – 3 day old biofilm grown in flow-cell stained with PI. C – GFP-tagged planktonic cells (OD – 0.5) stained with PI. Length of size bar: 15 µm.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>381747, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DNA_content_in_P_aeruginosa_cultures_/381747", "title"=>"DNA content in <i>P. aeruginosa</i> cultures.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:33:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/711499"], "description"=>"<p>Stationary cultures grown with 90 U/ml DNase I in the medium. After 48-h there was a clear difference of the visible aggregation in the culture. The control was left untreated. The top and bottom panels represent two independent experiments.</p>", "links"=>[], "tags"=>["dnase", "aggregate"], "article_id"=>381851, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g009"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_DNase_on_aggregate_formation_/381851", "title"=>"Effect of DNase on aggregate formation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:34:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/711606"], "description"=>"<p>We tested whether if mutants not able to produce the two distinct extracellular polysaccharides (Pel and Psl) could form intercellular fibers and if these fibers could be removed by DNAse treatment. WT - wild type PAO1, ΔpelA – PAO1 not able to produce Pel polysaccarides, ΔpslBCD - PAO1 not able to produce Psl polysaccarides, ΔpslBCDpelA - PAO1 not able to produce both Psl and Pel polysaccarides.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>381951, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g010"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Matrix_production_by_mutants_/381951", "title"=>"Matrix production by mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:34:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/711741"], "description"=>"<p>HHA-FITC and PI staining of aggregate harvested from a 48-h old stationary culture. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0027943#pone-0027943-g001\" target=\"_blank\">Figure 1</a> and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0027943#pone-0027943-g002\" target=\"_blank\">2</a> represent two independent experiments with A, B and C representing HHA-FITC, PI and HHA-FITC+PI. To test whether if the fibers are made from more than one polymer, we co-stained the aggregates with PI (red) and a mannose-specific lectin stain (HHA-FITC) to visualize DNA and any present Psl polymers. Length of size bar; <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0027943#pone-0027943-g001\" target=\"_blank\">Figure 1</a>: 5 µm; <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0027943#pone-0027943-g002\" target=\"_blank\">Figure 2</a>: 10 µm.</p>", "links"=>[], "tags"=>["lections", "dna"], "article_id"=>382089, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["Morten Alhede", "Kasper Nørskov Kragh", "Klaus Qvortrup", "Marie Allesen-Holm", "Maria van Gennip", "Louise D. Christensen", "Peter Østrup Jensen", "Anne K. Nielsen", "Matt Parsek", "Dan Wozniak", "Søren Molin", "Tim Tolker-Nielsen", "Niels Høiby", "Michael Givskov", "Thomas Bjarnsholt"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027943.g011"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Localization_of_lections_and_DNA_in_aggregates_/382089", "title"=>"Localization of lections and DNA in aggregates.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 11:35:45"}

PMC Usage Stats | Further Information

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Relative Metric

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