BiofilmPub Date : 2024-11-30eCollection Date: 2024-12-01DOI: 10.1016/j.bioflm.2024.100241
Santosh Pandit, Mi-A Kim, Ji-Eun Jung, Hyeon-Mi Choi, Jae-Gyu Jeon
{"title":"Usnic acid brief exposure suppresses cariogenic properties and complexity of <i>Streptococcus mutans</i> biofilms.","authors":"Santosh Pandit, Mi-A Kim, Ji-Eun Jung, Hyeon-Mi Choi, Jae-Gyu Jeon","doi":"10.1016/j.bioflm.2024.100241","DOIUrl":"https://doi.org/10.1016/j.bioflm.2024.100241","url":null,"abstract":"<p><p>Bacterial biofilms are highly structured surface associated architecture of micro-colonies, which are strongly bonded with the exopolymeric matrix of their own synthesis. These exopolymeric substances, mainly exopolysaccharides (EPS) initially assist the bacterial adhesion and finally form a bridge over the microcolonies to protect them from environmental assaults and antimicrobial exposure. Bacterial cells in dental biofilm metabolize dietary carbohydrates and produce organic acids. The blanket of exopolysaccharides over the bacterial communities hinders the buffering by saliva, contributing to the initiation of tooth decay followed by the progression of dental caries. Considering the current interest towards the use of natural antimicrobial agents to disarm the cariogenic properties of dental biofilm, this study evaluated the antimicrobial activity and the effect of twice daily brief exposure (1 min) of usnic acid on acid production, acid tolerance and development of 3-dimensional architecture of <i>Streptococcus mutans</i> biofilm. Herein, biofilms were briefly treated twice daily during biofilm development and biofilms were analyzed by using biochemical, microbiological and microscopic examination. Results obtained in this study showed a significant reduction in virulence properties of biofilm cells treated with usnic acid in compared to non-treated biofilms. Furthermore, twice daily brief exposure of usnic acid significantly disrupted the acid production and reduced the complexity of <i>Streptococcus mutans</i> biofilm by disrupting the EPS production. Brief exposure of usnic acid inhibited the production of glucosyltransferase (GTF) enzymes and their enzymatic activity leading to inhibition in production of EPS on the biofilm matrix. In conclusion, usnic acid treatment reduced the cariogenic properties and complexity of <i>S. mutans</i> biofilm by inhibiting acid production, acid tolerance and disrupting extracellular polysaccharide (EPS) formation, indicating its potential for preventing dental caries.</p>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"100241"},"PeriodicalIF":5.9,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11652789/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142856915","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-11-16DOI: 10.1016/j.bioflm.2024.100239
Jiyu Xie , Xinli Sun , Yanwei Xia , Lili Tao , Taimeng Tan , Nan Zhang , Weibing Xun , Ruifu Zhang , Ákos T. Kovács , Zhihui Xu , Qirong Shen
{"title":"Bridging the Gap: Biofilm-mediated establishment of Bacillus velezensis on Trichoderma guizhouense mycelia","authors":"Jiyu Xie , Xinli Sun , Yanwei Xia , Lili Tao , Taimeng Tan , Nan Zhang , Weibing Xun , Ruifu Zhang , Ákos T. Kovács , Zhihui Xu , Qirong Shen","doi":"10.1016/j.bioflm.2024.100239","DOIUrl":"10.1016/j.bioflm.2024.100239","url":null,"abstract":"<div><div>Bacterial-fungal interactions (BFIs) are important in ecosystem dynamics, especially within the soil rhizosphere. The bacterium <em>Bacillus velezensis</em> SQR9 and the fungus <em>Trichoderma guizhouense</em> NJAU 4742 have gathered considerable attention due to their roles in promoting plant growth and protecting their host against pathogens. In this study, we utilized these two model microorganisms to investigate BFIs. We firstly demonstrate that while co-inoculation of <em>B. velezensis</em> and <em>T. guizhouense</em> could promote tomato growth, these two microorganisms display mutual antagonism on agar solidified medium. To resolve this contradiction, we developed an inoculation method, that allows <em>B. velezensis</em> colonization of <em>T</em>. <em>guizhouense</em> hyphae and performed a transcriptome analysis. During colonization of the fungal hyphae, <em>B. velezensis</em> SQR9 upregulates expression of biofilm related genes (e.g. <em>eps, tasA</em>, and <em>bslA)</em> that is distinct from free-living cells. This result suggested an intricate association between extracellular matrix expression and hyphae colonization. In accordance, deletion <em>epsD</em>, <em>tasA, or</em> both <em>epsD</em> and <em>tasA</em> genes of <em>B. velezensis</em> diminished colonization of the <em>T</em>. <em>guizhouense</em> hyphae. The insights from our study demonstrate that soil BFIs are more complex than we understood, potentially involving both competition and cooperation. These intricate biofilm-mediated BFI dynamics might contribute to the remarkable diversity observed within soil microbiota, providing a fresh perspective for further exploration of BFIs in the plant rhizosphere.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100239"},"PeriodicalIF":5.9,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142706666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-11-16DOI: 10.1016/j.bioflm.2024.100240
Lucie Klopffer , Nicolas Louvet , Simon Becker , Jérémy Fix , Cédric Pradalier , Laurence Mathieu
{"title":"Effect of shear rate on early Shewanella oneidensis adhesion dynamics monitored by deep learning","authors":"Lucie Klopffer , Nicolas Louvet , Simon Becker , Jérémy Fix , Cédric Pradalier , Laurence Mathieu","doi":"10.1016/j.bioflm.2024.100240","DOIUrl":"10.1016/j.bioflm.2024.100240","url":null,"abstract":"<div><div>Understanding pioneer bacterial adhesion is essential to appreciate bacterial colonization and consider appropriate control strategies. This bacterial entrapment at the wall is known to be controlled by many physical, chemical or biological factors, including hydrodynamic conditions. However, due to the nature of early bacterial adhesion, i.e. a short and dynamic process with low biomass involved, such investigations are challenging. In this context, our study aimed to evaluate the effect of wall shear rate on the early bacterial adhesion dynamics. Firstly, at the population scale by assessing bacterial colonization kinetics and the mechanisms responsible for wall transfer under shear rates using a time-lapse approach. Secondly, at the individual scale, by implementing an automated image processing method based on deep learning to track each individual pioneer bacterium on the wall. Bacterial adhesion experiments are performed on a model bacterium (<em>Shewanella oneidensis</em> MR-1) at different shear rates (0 to1250 s<sup>−1</sup>) in a microfluidic system mounted under a microscope equipped with a CCD camera. Image processing was performed using a trained neural network (YOLOv8), which allowed information extraction, i.e. bacterial wall residence time and orientation for each adhered bacterium during pioneer colonization (14 min). Collected from over 20,000 bacteria, our results showed that adhered bacteria had a very short residence time at the wall, with over 70 % remaining less than 1 min. Shear rates had a non-proportional effect on pioneer colonization with a bell-shape profile suggesting that intermediate shear rates improved both bacterial wall residence time as well as colonization rate and level. This lack of proportionality highlights the dual effect of wall shear rate on early bacterial colonization; initially increasing it improves bacterial colonization up to a threshold, beyond which it leads to higher bacterial wall detachment. The present study provides quantitative data on the individual dynamics of just adhered bacteria within a population when exposed to different rates of wall shear.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100240"},"PeriodicalIF":5.9,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142706667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Small cyclic dipeptide produced by Lactobacillus rhamnosus with anti-biofilm properties against Streptococcus mutans biofilm","authors":"Rashmi Niranjan , Sachin Patil , Amrita Dubey , Bimlesh Lochab , Richa Priyadarshini","doi":"10.1016/j.bioflm.2024.100237","DOIUrl":"10.1016/j.bioflm.2024.100237","url":null,"abstract":"<div><div>The human oral cavity harbors many bacterial species collectively termed the oral microbiome and is integral for maintaining oral health. Dysbiosis of oral microbiota leads to common oral diseases, including dental caries, gingivitis, and periodontitis. <em>Streptococcus mutans</em> is the primary causative agent of dental caries. Studies have explored the use of probiotic <em>Lactobacillus</em> spp. to mitigate <em>S. mutans</em> biofilms. In the present study, we have tested the use of <em>Lactobacillus rhamnosus</em> extracts/metabolites for anti-biofilm properties. A small organic compound/metabolite was isolated from the cell-free supernatant of <em>L. rhamnosus</em>, and this metabolite resulted in a dose-dependent inhibition of <em>S. mutans</em> biofilms. Confocal microscopy revealed that the thickness of <em>S. mutans</em> biofilms was severely reduced upon metabolite treatment. With the help of FTIR spectra and mass spectrometry analysis, the molecular formula (C<sub>11</sub>H<sub>19</sub>O<sub>2</sub>N<sub>2</sub>) was deduced. The inhibitor compound was further identified as a small cyclic peptide, cyclo (-L-Leu-L-Pro). Our data also revealed that isolated metabolite impedes <em>S. mutans</em> biofilms by modulating gene expression of several essential genes involved in biofilm establishment.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100237"},"PeriodicalIF":5.9,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142661144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-11-06DOI: 10.1016/j.bioflm.2024.100238
Alicia Gomez-Lopez , Candela Fernandez-Fernandez
{"title":"Molecular characterization of gliotoxin synthesis in a biofilm model of Aspergillus fumigatus","authors":"Alicia Gomez-Lopez , Candela Fernandez-Fernandez","doi":"10.1016/j.bioflm.2024.100238","DOIUrl":"10.1016/j.bioflm.2024.100238","url":null,"abstract":"<div><div>Mycelial growth as biofilm structures and the activation of secondary metabolism leading to the release of low-molecular-weight molecules (known as secondary metabolites), are among the previously described strategies used by the filamentous fungi <em>Aspergillus fumigatus</em> to adapt and survive. Our study unveils that <em>A. fumigatus</em> strains can activate mechanisms linked to the production of gliotoxin, a crucial metabolite for <em>Aspergillus</em>, in the established <em>in vitro</em> biofilm model. Gliotoxin production exhibits strain- and time-dependent patterns and is associated -in a coordinated manner-with the expression levels of several genes involved in its regulation and synthesis. The transcriptional study of some of these genes by qPCR shows temporal inter-strain differences, which correlate with those obtained when evaluating the amounts of metabolites produced. Given that <em>A. fumigatus</em> forms biofilm structures within the site of infection, understanding the regulation of gliotoxin biosynthesis may have a role in the evolution of <em>Aspergillus</em> infection and guide diagnostic and treatment strategies.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100238"},"PeriodicalIF":5.9,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142661142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-11-01DOI: 10.1016/j.bioflm.2024.100236
Torgny Sunnerhagen , Thomas Bjarnsholt , Klaus Qvortrup , Henning Bundgaard , Claus Moser
{"title":"Transcatheter aortic valve implantation (TAVI) prostheses in vitro - biofilm formation and antibiotic effects","authors":"Torgny Sunnerhagen , Thomas Bjarnsholt , Klaus Qvortrup , Henning Bundgaard , Claus Moser","doi":"10.1016/j.bioflm.2024.100236","DOIUrl":"10.1016/j.bioflm.2024.100236","url":null,"abstract":"<div><h3>Background</h3><div>Transcatheter aortic valve implantation (TAVI) is a percutaneous catheter-based treatment of aortic stenosis as an alternative to open heart valve surgery. In cases of TAVI endocarditis, the treatment possibilities may be limited as surgical removal of the infected valve may be associated with a high risk in elderly, comorbid or frail patients. The propensity of bacteria to form a biofilm on foreign material is assumed to be of importance part of the disease process in TAVI endocarditis, but no studies on biofilm formation on TAVI valves have been conducted. We hypothesize that <em>Staphylococcus aureus</em> and <em>Enterococcus faecalis</em> biofilm formation on TAVI valves may have an impact on antibiotic tolerance and non-surgical cure rates.</div></div><div><h3>Methods</h3><div>TAVI valves (pieces including part of the metal frame, approximately 1 cm wide) were exposed to either species <em>in vitro</em> in LB-Krebs Ringer medium at 37 °C, with the bacterial count being assessed by culturing of sonicated TAVI pieces and broth at 0, 4, 18 and 24 h after bacterial exposure. Scanning electron microscopy (SEM) was performed. Effects of ampicillin, gentamicin, moxifloxacin, rifampicin (for <em>S. aureus</em>), and ceftriaxone (for <em>E. faecalis</em>) at 5 times minimal inhibitory concentration were tested alone and in combination with ampicillin. Antibiotics were added to biofilm aged 0 or 24 h and the effects assessed.</div></div><div><h3>Results</h3><div>Exposure for 15 min established attachment to all of valve pieces. SEM findings were consistent with biofilm formation and suggested lower amounts of bacteria on the metal compared to the tissue part of the TAVI valves. The number of bacteria attached to the TAVI valves increased until 24 h of incubation from less than 10^1 to a level of approximately 10^9 CFU/g. The bacteria became more tolerant to antibiotics on the TAVI valves over time, with the bactericidal effect against 24-h old biofilm being significantly less effective than against 0-h old biofilm depending on antibiotic.</div></div><div><h3>Conclusions</h3><div>The results indicate that bacteria can adhere to metal and tissue parts of the TAVI valves within minutes after an exposure which is comparable to transient bacteremia <em>in vivo</em>, and that the bacteria rapidly gain biofilm properties, associated with significantly reduced antibiotic effect.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100236"},"PeriodicalIF":5.9,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142573126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-10-24DOI: 10.1016/j.bioflm.2024.100234
Alfonz Kedves , Zoltán Kónya
{"title":"Effects of nanoparticles on anaerobic, anammox, aerobic, and algal-bacterial granular sludge: A comprehensive review","authors":"Alfonz Kedves , Zoltán Kónya","doi":"10.1016/j.bioflm.2024.100234","DOIUrl":"10.1016/j.bioflm.2024.100234","url":null,"abstract":"<div><div>Nanoparticles (NPs) are of significant interest due to their unique properties, such as large surface area and high reactivity, which have facilitated advancements in various fields. However, their increased use raises concerns about environmental impacts, including on wastewater treatment processes. This review examines the effects of different nanoparticles on anaerobic, anammox, aerobic, and algal-bacterial granular sludge used in wastewater treatment. CeO<sub>2</sub> and Ag NPs demonstrated adverse effects on aerobic granular sludge (AGS), reducing nutrient removal and cellular function, while anaerobic granular sludge (AnGS) and anammox granular sludge (AxGS) showed greater resilience due to their higher extracellular polymeric substance (EPS) content. TiO<sub>2</sub> NPs had fewer negative effects on algal-bacterial granular sludge (ABGS) than on AGS, as algae played a crucial role in enhancing EPS production and stabilizing the granules. The addition of Fe<sub>3</sub>O<sub>4</sub> NPs significantly enhanced both aerobic and anammox granulation by reducing granulation time, promoting microbial interactions, improving granule stability, and increasing nitrogen removal efficiency, primarily through increased EPS production and enzyme activity. However, Cu and CuO NPs exhibited strong inhibitory effects on aerobic, anammox, and anaerobic systems, affecting EPS structure, cellular integrity, and microbial viability. ZnO NPs demonstrated dose-dependent toxicity, with higher concentrations inducing oxidative stress and reducing performance in AGS and AnGS, whereas AxGS and ABGS were more tolerant due to enhanced EPS production and algae-mediated protection. The existing knowledge gaps and directions for future research on NPs are identified and discussed.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100234"},"PeriodicalIF":5.9,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142538350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-10-24DOI: 10.1016/j.bioflm.2024.100235
Matthew R. Willett , Sarah L. Codd , Joseph D. Seymour , Catherine M. Kirkland
{"title":"Relaxation-weighted MRI analysis of biofilm EPS: Differentiating biopolymers, cells, and water","authors":"Matthew R. Willett , Sarah L. Codd , Joseph D. Seymour , Catherine M. Kirkland","doi":"10.1016/j.bioflm.2024.100235","DOIUrl":"10.1016/j.bioflm.2024.100235","url":null,"abstract":"<div><div>Biofilms are a highly complex community of microorganisms embedded in a protective extracellular polymeric substance (EPS). Successful biofilm control requires a variety of approaches to better understand the structure-function relationship of the EPS matrix. Magnetic resonance imaging (MRI) is a versatile tool which can measure spatial structure, diffusion, and flow velocities in three dimensions and in situ. It is well-suited to characterize biofilms under natural conditions and at different length scales. MRI contrast is dictated by <em>T</em><sub><em>1</em></sub> and <em>T</em><sub><em>2</em></sub> relaxation times which vary spatially depending on the local chemical and physical environment of the sample. Previous studies have demonstrated that MRI can provide important insights into the internal structure of biofilms, but the contribution of major biofilm components—such as proteins, polysaccharides, and cells—to MRI contrast is not fully understood. This study explores how these components affect contrast in <em>T</em><sub><em>1</em></sub><em>-</em>and <em>T</em><sub><em>2</em></sub>-weighted MRI by analyzing artificial biofilms with well-defined properties modeled after aerobic granular sludge (AGS), compact spherical biofilm aggregates used in wastewater treatment. MRI of these biofilm models showed that certain gel-forming polysaccharides are a major source of <em>T</em><sub><em>2</em></sub> contrast, while other polysaccharides show minimal contrast. Proteins were found to reduce <em>T</em><sub><em>2</em></sub> contrast slightly when combined with polysaccharides, while cells had a negligible impact on <em>T</em><sub><em>2</em></sub> but showed <em>T</em><sub><em>1</em></sub> contrast. Patterns observed in the model biofilms served as a reference for examining <em>T</em><sub><em>2</em></sub> and <em>T</em><sub><em>1</em></sub>-weighted contrast in the void spaces of two distinct AGS granules, allowing for a qualitative evaluation of the EPS components which may be present. Further insights provided by MRI may help improve understanding of the biofilm matrix and guide how to better manage biofilms in wastewater, clinical, and industrial settings.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100235"},"PeriodicalIF":5.9,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142661143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-10-24DOI: 10.1016/j.bioflm.2024.100232
Jung-Ah Cho , Sangsoo Jeon , Youngmin Kwon , Yoo Jin Roh , Sukjin Shin , Chang-Hun Lee , Sung Jae Kim
{"title":"Identification and comparison of protein composition of biofilms in response to EGCG from Enterococcus faecalis and Staphylococcus lugdunensis, which showed opposite patterns in biofilm-forming abilities","authors":"Jung-Ah Cho , Sangsoo Jeon , Youngmin Kwon , Yoo Jin Roh , Sukjin Shin , Chang-Hun Lee , Sung Jae Kim","doi":"10.1016/j.bioflm.2024.100232","DOIUrl":"10.1016/j.bioflm.2024.100232","url":null,"abstract":"<div><div>Bacterial biofilm is resistant to conventional antibiotic treatments, leading to complications associated with many infection-related human diseases. Epigallocatechin Gallate (EGCG), a phenolic catechin enriched in green tea, is recognized for its anti-bacterial and anti-biofilm activities. In this study, we examined the protein components of the biofilms formed in the absence or presence of EGCG using <em>Enterococcus faecalis</em> and <em>Staphylococcus lugdunensis,</em> which had shown opposing patterns in biofilm formation. A clustering heatmap revealed that the two microorganisms expressed the different protein sets in response to EGCG. Proteins that were noticeably upregulated included those associated with stress responsiveness and gluconeogenesis in <em>E. faecalis</em>, and gene modification in <em>S. lugdunensis</em>. Conversely, downregulated proteins were related to tRNA-modifying enzyme activity in <em>E. faecalis</em>, and anabolic metabolism in <em>S. lugdunensis</em>. Among the proteins identified only in EGCG-responsive biofilms, enzymes involved in de novo purine biosynthesis were enriched in <em>E. faecalis</em>, while proteins likely to cause DNA instability and pathogenicity changes were abundantly present in <em>S. lugdunensis</em>. The classification based on gene ontology (GO) terms by microorganism exhibited that metabolic process or catabolic activity was at the top rank in <em>E. faecalis</em> with more than 33 proteins, and in <em>S. lugdunensis</em>, localization or transport was highly ranked with 4 proteins. These results support the hypothesis that EGCG might cause different cellular programs in each microorganism. Finally, comparison of the proteomes between two groups that form biofilms to similar extents discovered that 2 proteins were commonly found in the weak biofilm-forming groups (<em>E. faecalis</em> and EGCG-responding <em>S. lugudunensis</em>), whereas 9 proteins were common among the strong biofilm-forming groups (<em>S. lugdunensis</em> and EGCG-responding <em>E. faecalis</em>). It was suggested that these proteins could serve as potential indicators to detect the presence and predict the extent of biofilm formation by multiple microorganisms. Taken all together, proteomics data and analyses performed in this study provided useful and new information on the proteins embedded in the biofilms formed at the specific conditions, which can aid in diagnosis and the development of tailored treatment strategies.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100232"},"PeriodicalIF":5.9,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142560756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BiofilmPub Date : 2024-10-23DOI: 10.1016/j.bioflm.2024.100233
Dominique C.S. Evans , Amanda B. Khamas , Alex Payne-Dwyer , Adam J.M. Wollman , Kristian S. Rasmussen , Janne K. Klitgaard , Birgitte Kallipolitis , Mark C. Leake , Rikke L. Meyer
{"title":"Cooperation between coagulase and von willebrand factor binding protein in Staphylococcus aureus fibrin pseudocapsule formation","authors":"Dominique C.S. Evans , Amanda B. Khamas , Alex Payne-Dwyer , Adam J.M. Wollman , Kristian S. Rasmussen , Janne K. Klitgaard , Birgitte Kallipolitis , Mark C. Leake , Rikke L. Meyer","doi":"10.1016/j.bioflm.2024.100233","DOIUrl":"10.1016/j.bioflm.2024.100233","url":null,"abstract":"<div><div>The major human pathogen <em>Staphylococcus aureus</em> forms biofilms comprising of a fibrin network that increases attachment to surfaces and shields bacteria from the immune system. It secretes two coagulases, Coagulase (Coa) and von Willebrand factor binding protein (vWbp), which hijack the host coagulation cascade and trigger the formation of this fibrin clot. However, it is unclear how Coa and vWbp contribute differently to the localisation and dynamics of clot assembly in growing biofilms.</div><div>Here, we address this question using high-precision time-resolved confocal microscopy of fluorescent fibrin to establish the spatiotemporal dynamics of fibrin clot formation in functional biofilms. We also use fluorescent fusion proteins to visualise the locations of Coa and vWbp in biofilms using both confocal laser scanning and high resolution highly inclined and laminated optical sheet microscopy. We visualise and quantify the spatiotemporal dynamics of fibrin production during initiation of biofilms in plasma amended with fluorescently labelled fibrinogen.</div><div>We find that human serum stimulates coagulase production, and that Coa and vWbp loosely associate to the bacterial cell surface. Coa localises to cell surfaces to produce a surface-attached fibrin pseudocapsule but can diffuse from cells to produce matrix-associated fibrin. vWbp produces matrix-associated fibrin in the absence of Coa, and furthermore accelerates pseudocapsule production when Coa is present. Finally, we observe that fibrin production varies across the biofilm. A sub-population of non-dividing cells does not produce any pseudocapsule but remains within the protective extended fibrin network, which could be important for the persistence of <em>S. aureus</em> biofilm infections as antibiotics are more effective against actively growing cells.</div><div>Our findings indicate a more cooperative role between Coa and vWbp in building fibrin networks than previously thought, and a bet-hedging cell strategy where some cells produce biofilm matrix while others do not, but instead assume a dormant phenotype that could be associated with antibiotic tolerance.</div></div>","PeriodicalId":55844,"journal":{"name":"Biofilm","volume":"8 ","pages":"Article 100233"},"PeriodicalIF":5.9,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142554271","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}