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Microbial Biofilms: Mechanisms of Formation, Pathogenicity and Antibiotic Resistance

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A special issue of Pathogens (ISSN 2076-0817).

Deadline for manuscript submissions: closed (30 September 2021) | Viewed by 20959

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Special Issue Editors

Prof. Dr. Patricia Poeta

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Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal
Interests: microbiology; one health; antimicrobial resistance; biofilms; microbial genetics; infectious diseases
Special Issues, Collections and Topics in MDPI journals

Dr. Vanessa Silva

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Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal
Interests: microbiology; one health; antimicrobial resistance; biofilms; microbial genetics; infectious diseases
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Dr. Carla Miranda

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1. Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal
2. LAQV@REQUIMTE, Chemistry Department, Faculty of Science and Technology, NOVA University of Lisbon, Almada, Portugal
Interests: microbiology; One Health; antimicrobial resistance; biofilms; microbial genetics; infectious diseases
Special Issues, Collections and Topics in MDPI journals

Dr. Gilberto Igrejas

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Special Issue Information

Dear Colleagues,

Biofilms are structured aggregates of bacterial cells that are embedded in self-produced extracellular polymeric substances. Biofilm formation occurs to enable bacterial survival, which requires physiological changes. Nearly 80% of all human infections are biofilm-related, and one of their most critical features is their considerably higher resistance to environmental stresses, antimicrobials, disinfectants and host immune defenses. Moreover, when antibiotic-resistant bacteria form a biofilm, the overall resistance is enhanced. Despite major advances in biofilm research, knowledge on biofilm formation, propagation and resistance is still very limited, and this poor understanding has hampered the development of antimicrobial drugs that specifically target biofilms. In fact, most of these studies have been focused on bacteria growing in planktonic cultures and hence have overlooked biofilm-specific AMR mechanisms. These are known to be distinct from the well-characterized intrinsic mechanisms that occur at the cellular level, being operated additively to the intrinsic mechanisms, in a transient and reversible manner, resulting in up to 1000-fold higher resistance levels. This Special Issue will bring together the latest studies regarding the mechanisms of biofilm formation as well as the aspects of function and adhesion that are associated with their pathogenicity. Furthermore, it will highlight the mechanisms of bacterial biofilm resistance to antimicrobial agents.

Prof. Dr. Patricia Poeta
Dr. Vanessa Silva
Prof. Dr. Carla Miranda
Prof. Dr. Gilberto Igrejas
Guest Editors

Manuscript Submission Information

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Keywords

biofilms
pathogenicity
environmental stresses
antimicrobials
disinfectants

Published Papers (5 papers)

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Research

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18 pages, 5348 KiB

Open AccessEditor’s ChoiceArticle

Biofilm Formation of Multidrug-Resistant MRSA Strains Isolated from Different Types of Human Infections

by Vanessa Silva, Luciana Almeida, Vânia Gaio, Nuno Cerca, Vera Manageiro, Manuela Caniça, José L. Capelo, Gilberto Igrejas and Patrícia Poeta

Pathogens 2021, 10(8), 970; https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens10080970 - 30 Jul 2021

Cited by 29 | Viewed by 4698

Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the main pathogens causing chronic infections, mainly due to its capacity to form biofilms. However, the mechanisms underlying the biofilm formation of MRSA strains from different types of human infections are not fully understood. MRSA strains isolated from distinct human infections were characterized aiming to determine their biofilm-forming capacity, the biofilm resistance to conventional antibiotics and the prevalence of biofilm-related genes, including, icaA, icaB, icaC, icaD, fnbA, fnbB, clfA, clfB, cna, eno, ebpS, fib and bbp. Eighty-three clinical MRSA strains recovered from bacteremia episodes, osteomyelitis and diabetic foot ulcers were used. The biofilm-forming capacity was evaluated by the microtiter biofilm assay and the biofilm structure was analyzed via confocal scanning laser microscopy. The antimicrobial susceptibility of 24-h-old biofilms was assessed against three antibiotics and the biomass reduction was measured. The metabolic activity of biofilms was evaluated by the XTT assay. The presence of biofilm-related genes was investigated by whole-genome sequencing and by PCR. Despite different intensities, all strains showed the capacity to form biofilms. Most strains had also a large number of biofilm-related genes. However, strains isolated from osteomyelitis showed a lower capacity to form biofilms and also a lower prevalence of biofilm-associated genes. There was a significant reduction in the biofilm biomass of some strains tested against antibiotics. Our results provide important information on the biofilm-forming capacity of clinical MRSA strains, which may be essential to understand the influence of different types of infections on biofilm production and chronic infections. Full article

(This article belongs to the Special Issue Microbial Biofilms: Mechanisms of Formation, Pathogenicity and Antibiotic Resistance)

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22 pages, 4918 KiB

Open AccessArticle

Activity of Liquid and Volatile Fractions of Essential Oils against Biofilm Formed by Selected Reference Strains on Polystyrene and Hydroxyapatite Surfaces

by Ruth Dudek-Wicher, Justyna Paleczny, Beata Kowalska-Krochmal, Patrycja Szymczyk-Ziółkowska, Natalia Pachura, Antoni Szumny and Malwina Brożyna

Pathogens 2021, 10(5), 515; https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens10050515 - 23 Apr 2021

Cited by 6 | Viewed by 2605

Abstract

Biofilms are surface-attached, structured microbial communities displaying higher tolerance to antimicrobial agents in comparison to planktonic cells. An estimated 80% of all infections are thought to be biofilm-related. The drying pipeline of new antibiotics efficient against biofilm-forming pathogens urges the search for alternative routes of treatment. Essential Oils (EOs), extracted from medicinally important plants, are a reservoir of bioactive compounds that may serve as a foothold in investigating novel antibiofilm compounds. The aim of this study was to compare antimicrobial activity of liquid and volatile fractions of tested EOs against biofilm-forming pathogens using different techniques. In this research, we tested five EOs, extracted from Syzygium aromaticum L., Boswelia serrata Roxb., Juniperus virginiana L., Pelargonium graveolens L. and Melaleuca alternifolia Cheel., against planktonic and biofilm forms of five selected reference strains, namely Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans. To obtain cohesive results, we applied four various methodological approaches: to assess the activity of the liquid fraction of EOs, disc diffusion and the microdilution method were applied; to test EOs’ volatile fraction, the AntiBioVol assay and modified Antibiofilm Dressing Activity Measurement (A.D.A.M.) were used. The molecular composition and dynamics of antimicrobial substances released from specific EOs was measured using Gas Chromatography–Mass Spectrometry (GC-MS). The antimicrobial potency of EO’s volatile fraction against biofilm formed by tested strains differed from that of the liquid fraction and was related to the molecular weight of volatile compounds. The liquid fraction of CW-EO and volatile fraction of F-EO acted in the strongest manner against biofilm of C. albicans. The addition of 0.5% Tween 20 to liquid phase, enhanced activity of G-EO against E. coli and K. pneumoniae biofilm. EO activity depended on the microbial species it was applied against and the chosen assessment methodology. While all tested EOs have shown a certain level of antimicrobial and antibiofilm effect, our results indicate that the choice of EO to be applied against a specific biofilm-forming pathogen requires careful consideration with regard to the above-listed aspects. Nevertheless, the results presented in this research contribute to the growing body of evidence indicating the beneficial effects of EOs, which may be applied to fight biofilm-forming pathogens. Full article

(This article belongs to the Special Issue Microbial Biofilms: Mechanisms of Formation, Pathogenicity and Antibiotic Resistance)

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14 pages, 1693 KiB

Open AccessArticle

Cytotoxicity, Intracellular Replication, and Contact-Dependent Pore Formation of Genotyped Environmental Legionella pneumophila Isolates from Hospital Water Systems in the West Bank, Palestine

by Ashraf R. Zayed, Marina Pecellin, Lina Jaber, Suha Butmeh, Shereen A. Bahader, Michael Steinert, Manfred G. Höfle, Ingrid Brettar and Dina M. Bitar

Pathogens 2021, 10(4), 417; https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens10040417 - 01 Apr 2021

Cited by 4 | Viewed by 2340

Abstract

Legionella pneumophila is the causative agent of Legionnaires’ disease. Due to the hot climate and intermittent water supply, the West Bank, Palestine, can be considered a high-risk area for this often fatal atypical pneumonia. L. pneumophila occurs in biofilms of natural and man-made freshwater environments, where it infects and replicates intracellularly within protozoa. To correlate the genetic diversity of the bacteria in the environment with their virulence properties for protozoan and mammalian host cells, 60 genotyped isolates from hospital water systems in the West Bank were analyzed. The L. pneumophila isolates were previously genotyped by high resolution Multi Locus Variable Number of Tandem Repeat Analysis (MLVA-8(12)) and sorted according to their relationship in clonal complexes (VACC). Strains of relevant genotypes and VACCs were compared according to their capacity to infect Acanthamoeba castellanii and THP-1 macrophages, and to mediate pore-forming cytotoxicity in sheep red blood cells (sRBCs). Based on a previous detailed analysis of the biogeographic distribution and abundance of the MLVA-8(12)-genotypes, the focus of the study was on the most abundant L. pneumophila- genotypes Gt4(17), Gt6 (18) and Gt10(93) and the four relevant clonal complexes [VACC1, VACC2, VACC5 and VACC11]. The highly abundant genotypes Gt4(17) and Gt6(18) are affiliated with VACC1 and sequence type (ST)1 (comprising L. pneumophila str. Paris), and displayed seroroup (Sg)1. Isolates of these two genotypes exhibited significantly higher virulence potentials compared to other genotypes and clonal complexes in the West Bank. Endemic for the West Bank was the clonal complex VACC11 (affiliated with ST461) represented by three relevant genotypes that all displayed Sg6. These genotypes unique for the West Bank showed a lower infectivity and cytotoxicity compared to all other clonal complexes and their affiliated genotypes. Interestingly, the L. pneumophila serotypes ST1 and ST461 were previously identified by in situ-sequence based typing (SBT) as main causative agents of Legionnaires’ disease (LD) in the West Bank at a comparable level. Overall, this study demonstrates the site-specific regional diversity of L. pneumophila genotypes in the West Bank and suggests that a combination of MLVA, cellular infection assays and hierarchical agglomerative cluster analysis allows an improved genotype-based risk assessment. Full article

(This article belongs to the Special Issue Microbial Biofilms: Mechanisms of Formation, Pathogenicity and Antibiotic Resistance)

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18 pages, 4623 KiB

Open AccessArticle

Identification and Morphological Characterization of Biofilms Formed by Strains Causing Infection in Orthopedic Implants

by Débora C. Coraça-Huber, Lisa Kreidl, Stephan Steixner, Maximilian Hinz, Dietmar Dammerer and Manfred Fille

Pathogens 2020, 9(8), 649; https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens9080649 - 12 Aug 2020

Cited by 13 | Viewed by 3468

Abstract

Objectives: For a better understanding of the mechanisms involved in biofilm formation, we performed a broad identification and characterization of the strains affecting implants by evaluating the morphology of biofilms formed in vitro in correlation with tests of the strains’ antibiotic susceptibility in planktonic form. The ability of the strains to form biofilms in vitro was evaluated by means of colony forming units counting, metabolic activity tests of biofilm cells, and scanning electron microscopy. Methods: A total of 140 strains were isolated from patients with orthopedic implant-related infections during the period of 2015 to 2018. The identification of the isolates was carried out through microbiological cultures and confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Antibiotic susceptibility rates of the isolates were accessed according to EUCAST (European Committee on Antimicrobial Susceptibility Testing). The ability of all isolates to form biofilms in vitro was evaluated by counting the colony forming units, by measuring the metabolic activity of biofilm cells, and by analyzing the morphology of the formed biofilms using scanning electron microscopy. Results: From all the isolates, 41.84% (62 strains) were Staphylococcus epidermidis and 15.60% (22 strains) were Staphylococcus aureus. A significant difference in the capacity of biofilm formation was observed among the isolates. When correlating the biofilm forming capacity of the isolates to their antibiotic susceptibility rates, we observed that not all strains that were classified as resistant were biofilm producers in vitro. In other words, bacteria that are not good biofilm formers can show increased tolerance to multiple antibiotic substances. Conclusion: From 2015 until 2018, Staphylococcus epidermidis was the strain that caused most of the orthopedic implant-related infections in our hospital. Not all strains causing infection in orthopedic implants are able to form biofilms under in vitro conditions. Differences were observed in the number of cells and morphology of the biofilms. In addition, antibiotic resistance is not directly related to the capacity of the strains to form biofilms in vitro. Further studies should consider the use of in vitro culture conditions that better reproduce the joint environment and the growth of biofilms in humans. Full article

(This article belongs to the Special Issue Microbial Biofilms: Mechanisms of Formation, Pathogenicity and Antibiotic Resistance)

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Review

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20 pages, 10165 KiB

Open AccessEditor’s ChoiceReview

Biofilm Formation as a Complex Result of Virulence and Adaptive Responses of Helicobacter pylori

by Paweł Krzyżek, Rossella Grande, Paweł Migdał, Emil Paluch and Grażyna Gościniak

Pathogens 2020, 9(12), 1062; https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens9121062 - 18 Dec 2020

Cited by 40 | Viewed by 6589

Abstract

Helicobacter pylori is a bacterium that is capable of colonizing a host for many years, often for a lifetime. The survival in the gastric environment is enabled by the production of numerous virulence factors conditioning adhesion to the mucosa surface, acquisition of nutrients, and neutralization of the immune system activity. It is increasingly recognized, however, that the adaptive mechanisms of H. pylori in the stomach may also be linked to the ability of this pathogen to form biofilms. Initially, biofilms produced by H. pylori were strongly associated by scientists with water distribution systems and considered as a survival mechanism outside the host and a source of fecal-oral infections. In the course of the last 20 years, however, this trend has changed and now the most attention is focused on the biomedical aspect of this structure and its potential contribution to the therapeutic difficulties of H. pylori. Taking into account this fact, the aim of the current review is to discuss the phenomenon of H. pylori biofilm formation and present this mechanism as a resultant of the virulence and adaptive responses of H. pylori, including morphological transformation, membrane vesicles secretion, matrix production, efflux pump activity, and intermicrobial communication. These mechanisms will be considered in the context of transcriptomic and proteomic changes in H. pylori biofilms and their modulating effect on the development of this complex structure. Full article

(This article belongs to the Special Issue Microbial Biofilms: Mechanisms of Formation, Pathogenicity and Antibiotic Resistance)

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