Browsing by Subject "biofilms"

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  • Langenheder, Silke; Wang, Jianjun; Karjalainen, Satu Maaria; Laamanen, Tiina M.; Tolonen, Kimmo T.; Vilmi, Annika; Heino, Jani (2017)
    The spatial structure and underlying assembly mechanisms of bacterial communities have been studied widely across aquatic systems, focusing primarily on isolated sites, such as different lakes, ponds and streams. Here, our main aim was to determine the underlying mechanisms for bacterial biofilm assembly within a large, highly connected lake system in Northern Finland using associative methods based on taxonomic and phylogenetic alpha-and beta-diversity and a large number of abiotic and biotic variables. Furthermore, null model approaches were used to quantify the relative importance of different community assembly processes. We found that spatial variation in bacterial communities within the lake was structured by different assembly processes, including stochasticity, species sorting and potentially even dispersal limitation. Species sorting by abiotic environmental conditions explained more of the taxonomic and particularly phylogenetic turnover in community composition compared with that by biotic variables. Finally, we observed clear differences in alpha diversity (species richness and phylogenetic diversity), which were to a stronger extent determined by abiotic compared with biotic factors, but also by dispersal effects. In summary, our study shows that the biodiversity of bacterial biofilm communities within a lake ecosystem is driven by within-habitat gradients in abiotic conditions and by stochastic and deterministic dispersal processes.
  • Nowak, Jessika; Visnovsky, Sandra B.; Pitman, Andrew R.; Cruz, Cristina D.; Palmer, Jon; Fletcher, Graham C.; Flint, Steve (2021)
    Listeria monocytogenes is a ubiquitous foodborne pathogen that results in a high rate of mortality in sensitive and immunocompromised people. Contamination of food with L. monocytogenes is thought to occur during food processing, most often as a result of the pathogen producing a biofilm that persists in the environment and acting as the source for subsequent dispersal of cells onto food. A survey of seafoodprocessing plants in New Zealand identified the persistent strain 15G01, which has a high capacity to form biofilms. In this study, a transposon library of L. monocytogenes 15G01 was screened for mutants with altered biofilm formation, assessed by a crystal violet assay, to identify genes involved in biofilm formation. This screen identified 36 transposants that showed a significant change in biofilm formation compared to the wild type. The insertion sites were in 27 genes, 20 of which led to decreased biofilm formation and seven to an increase. Two insertions were in intergenic regions. Annotation of the genes suggested that they are involved in diverse cellular processes, including stress response, autolysis, transporter systems, and cell wall/membrane synthesis. Analysis of the biofilms produced by the transposants using scanning electron microscopy and fluorescence microscopy showed notable differences in the structure of the biofilms compared to the wild type. In particular, inactivation of uvrB and mltD produced coccoid-shaped cells and elongated cells in long chains, respectively, and the mgtB mutant produced a unique biofilm with a sandwich structure which was reversed to the wild-type level upon magnesium addition. The mltD transposant was successfully complemented with the wild-type gene, whereas the phenotypes were not or only partially restored for the remaining mutants. IMPORTANCE The major source of contamination of food with Listeria monocytogenes is thought to be due to biofilm formation and/or persistence in food-processing plants. By establishing as a biofilm, L. monocytogenes cells become harder to eradicate due to their increased resistance to environmental threats. Understanding the genes involved in biofilm formation and their influence on biofilm structure will help identify new ways to eliminate harmful biofilms in food processing environments. To date, multiple genes have been identified as being involved in biofilm formation by L. monocytogenes; however, the exact mechanism remains unclear. This study identified four genes associated with biofilm formation by a persistent strain. Extensive microscopic analysis illustrated the effect of the disruption of mgtB, clsA, uvrB, and mltD and the influence of magnesium on the biofilm structure. The results strongly suggest an involvement in biofilm formation for the four genes and provide a basis for further studies to analyze gene regulation to assess the specific role of these biofilm-associated genes.
  • Teirlinck, Eline; Barras, Alexandre; Liu, Jing; Fraire, Juan C.; Lajunen, Tatu; Xiong, Ranhua; Forier, Katrien; Li, Chengnan; Urtti, Arto; Boukherroub, Rabah; Szunerits, Sabine; De Smedt, Stefaan C.; Coenye, Tom; Braeckmans, Kevin (2019)
    Impaired penetration of antibiotics through bacterial biofilms is one of the reasons for failure of antimicrobial therapy. Hindered drug diffusion is caused on the one hand by interactions with the sticky biofilm matrix and on the other hand by the fact that bacterial cells are organized in densely packed clusters of cells. Binding interactions with the biofilm matrix can be avoided by encapsulating the antibiotics into nanocarriers, while interfering with the integrity of the dense cell clusters can enhance drug transport deep into the biofilm. Vapor nanobubbles (VNB), generated from laser irradiated nanoparticles, are a recently reported effective way to loosen up the biofilm structure in order to enhance drug transport and efficacy. In the present study, we explored if the disruptive force of VNB can be used simultaneously to interfere with the biofilm structure and trigger antibiotic release from light-responsive nanocarriers. The antibiotic tobramycin was incorporated in two types of light-responsive nanocarriersliposomes functionalized with gold nanoparticles (Lip-AuNP) and graphene quantum dots (GQD)and their efficacy was evaluated on Pseudomonas aeruginosa biofilms. Even though the anti-biofilm efficacy of tobramycin was improved by liposomal encapsulation, electrostatic functionalization with 70 nm AuNP unfortunately resulted in premature leakage of tobramycin in a matter of hours. Laser-irradiation consequently did not further improve P. aeruginosa biofilm eradication. Adsorption of tobramycin to GQD, on the other hand, did result in a stable formulation with high encapsulation efficiency, without burst release of tobramycin from the nanocarriers. However, even though laser-induced VNB formation from GQD resulted in biofilm disruption, an enhanced anti-biofilm effect was not achieved due to tobramycin not being efficiently released from GQD. Even though this study was unsuccessful in designing suitable nanocarriers for simultaneous biofilm disruption and light-triggered release of tobramycin, it provides insights into the difficulties and challenges that need to be considered for future developments in this regard.
  • Mosselhy, Dina A.; Assad, Mhd; Sironen, Tarja; Elbahri, Mady (2021)
    Staphylococcus aureus is a notorious pathogen that colonizes implants (orthopedic and breast implants) and wounds with a vicious resistance to antibiotic therapy. Methicillin-resistant S. aureus (MRSA) is a catastrophe mainly restricted to hospitals and emerged to community reservoirs, acquiring resistance and forming biofilms. Treating biofilms is problematic except via implant removal or wound debridement. Nanoparticles (NPs) and nanofibers could combat superbugs and biofilms and rapidly diagnose MRSA. Nanotheranostics combine diagnostics and therapeutics into a single agent. This comprehensive review is interpretative, utilizing mainly recent literature (since 2016) besides the older remarkable studies sourced via Google Scholar and PubMed. We unravel the molecular S. aureus resistance and complex biofilm. The diagnostic properties and detailed antibacterial and antibiofilm NP mechanisms are elucidated in exciting stories. We highlight the challenges of bacterial infections nanotheranostics. Finally, we discuss the literature and provide "three action appraisals". (i) The first appraisal consists of preventive actions (two wings), avoiding unnecessary hospital visits, hand hygiene, and legislations against over-the-counter antibiotics as the general preventive wing. Our second recommended preventive wing includes preventing the adverse side effects of the NPs from resistance and toxicity by establishing standard testing procedures. These standard procedures should provide breakpoints of bacteria's susceptibility to NPs and a thorough toxicological examination of every single batch of synthesized NPs. (ii) The second appraisal includes theranostic actions, using nanotheranostics to diagnose and treat MRSA, such as what we call "multifunctional theranostic nanofibers. (iii) The third action appraisal consists of collaborative actions.
  • Gilbert-Girard, Shella; Savijoki, Kirsi; Yli-Kauhaluoma, Jari; Fallarero, Adyary (2020)
    In recent years, bacterial infections have become a main concern following the spread of antimicrobial resistance. In addition, bacterial biofilms are known for their high tolerance to antimicrobials and they are regarded as a main cause of recalcitrant infections in humans. Many efforts have been deployed in order to find new antibacterial therapeutic options and the high-throughput screening (HTS) of large libraries of compounds is one of the utilized strategies. However, HTS efforts for anti-biofilm discovery remain uncommon. Here, we miniaturized a 96-well plate (96WP) screening platform, into a 384-well plate (384WP) format, based on a sequential viability and biomass measurements for the assessment of anti-biofilm activity. During the assay optimization process, different parameters were evaluated while using Staphylococcus aureus and Pseudomonas aeruginosa as the bacterial models. We compared the performance of the optimized 384WP platform to our previously established 96WP-based platform by carrying out a pilot screening of 100 compounds, followed by the screening of a library of 2000 compounds to identify new repurposed anti-biofilm agents. Our results show that the optimized 384WP platform is well-suited for screening purposes, allowing for the rapid screening of a higher number of compounds in a run in a reliable manner.
  • Munsch-Alatossava, Patricia; Alatossava, Tapani (2020)
    Worldwide, the dairy sector remains of vital importance for food production despite severe environmental constraints. The production and handling conditions of milk, a rich medium, promote inevitably the entrance of microbial contaminants, with notable impact on the quality and safety of raw milk and dairy products. Moreover, the persistence of high concentrations of microorganisms (especially bacteria and bacterial spores) in biofilms (BFs) present on dairy equipment or environments constitutes an additional major source of milk contamination from pre- to post-processing stages: in dairies, BFs represent a major concern regarding the risks of disease outbreaks and are often associated with significant economic losses. One consumption trend toward "raw or low-processed foods" combined with current trends in food production systems, which tend to have more automation and longer processing runs with simultaneously more stringent microbiological requirements, necessitate the implementation of new and obligatory sustainable strategies to respond to new challenges regarding food safety. Here, in light of studies, performed mainly with raw milk, that considered dominant "planktonic" conditions, we reexamine the changes triggered by cold storage alone or combined with nitrogen gas (N-2) flushing on bacterial populations and discuss how the observed benefits of the treatment could also contribute to limiting BF formation in dairies.
  • Savijoki, Kirsi; Myllymäki, Henna; Luukinen, Hanna; Paulamäki, Lauri; Vanha-aho, Leena-Maija; Svorjova, Aleksandra; Miettinen, Ilkka; Fallarero, Adyary; Ihalainen, Teemu O; Yli-Kauhaluoma, Jari; Nyman, Tuula A.; Parikka, Mataleena (2021)
    The complex cell wall and biofilm matrix (ECM) act as key barriers to anti-biotics in mycobacteria. Here, the ECM and envelope proteins ofMycobacterium mari-numATCC 927, a nontuberculous mycobacterial model, were monitored over 3 monthsby label-free proteomics and compared with cell surface proteins on planktonic cells touncover pathways leading to virulence, tolerance, and persistence. We show that ATCC927 forms pellicle-type and submerged-type biofilms (PBFs and SBFs, respectively) after2 weeks and 2 days of growth, respectively, and that the increased CelA1 synthesis inthis strain prevents biofilm formation and leads to reduced rifampicin tolerance. Theproteomic data suggest that specific changes in mycolic acid synthesis (cord factor),Esx1 secretion, and cell wall adhesins explain the appearance of PBFs as ribbon-likecords and SBFs as lichen-like structures. A subpopulation of cells resisting 64MICrifampicin (persisters) was detected in both biofilm subtypes and already in 1-week-oldSBFs. The key forces boosting their development could include subtype-dependentchanges in asymmetric cell division, cell wall biogenesis, tricarboxylic acid/glyoxylatecycle activities, and energy/redox/iron metabolisms. The effect of various ambient oxy-gen tensions on each cell type and nonclassical protein secretion are likely factorsexplaining the majority of the subtype-specific changes. The proteomicfindings alsoimply that Esx1-type protein secretion is more efficient in planktonic (PL) and PBF cells,while SBF may prefer both the Esx5 and nonclassical pathways to control virulence andprolonged viability/persistence. In conclusion, this study reports thefirst proteomicinsight into aging mycobacterial biofilm ECMs and indicates biofilm subtype-dependentmechanisms conferring increased adaptive potential and virulence of nontuberculousmycobacteria.
  • Manner, Suvi; Skogman, Malena; Goeres, Darla; Vuorela, Pia; Fallarero, Adyary (2013)
  • Ahola-Iivarinen, Elina (Helsingfors universitet, 2016)
    In the literature review of this study, the focus was on biofilms that certain microbes produce, and their potential use in food industry. Biofilms consist of microbial cells and extracellular products, e.g., polysaccharides. Pullulan as an exopolysaccharide has many industrial applications and the aim of this study was to explore a new potential alpha-glucan, dextran, and especially its ability to form a stand-alone film. Pullulan and dextran were separately mixed in de-ionized water. The dynamic viscosities of dextran and pullulan solutions were determined. Film formation of dextran was not successful, not even with sorbitol as a plasticizer. The optical properties, water vapor and oxygen permeabilities and tensile strengths of pullulan films were studied. Additionally, Whatman42-filter material was coated with or immersed in dextran solution. Hence the changes in tensile strength and permeability values between a well-known material and dextran treated material could be detected. Pullulan films had low haze values (2.1–3.9%) and they were transparent to UVA-, UVB- and visible light. The tensile strength values of pullulan films were 47–53 MPa. For filter paper, the corresponding values were 10 MPa and application of dextran coating increased it to 15–19 MPa. All polysaccharide solutions exhibited Newtonian behavior and their relative viscosities were <10 mPa, 5% pullulan with viscosity around 20 mPa as an exception. Pullulan solutions had higher viscosities than dextran solutions. The air permeabilities were 10–50 ml/min for pullulan films, 10 ml/min for dextran-sorbitol film, 200 ml/min for dextran film and 200–500 ml/min for Whatman42 material. The oxygen permeability values for pullulan films were <0,1 cm3·μm m-2·d-1·kPa-1. Based on results in this study, pullulan films are impermeable to oxygen. As the films tolerated water vapor poorly, pullulan might be a potential component in packages made of composite materials, as individual packaging material in dry environment or possibly chemically modified to obtain better resistance to water vapor. Our results show that without additional modifications dextran does not form a continuous self-supporting films in these conditions.
  • Dickinson, Amy; Kankaanpää, Hanna; Silen, Suvi; Meri, Seppo; Haapaniemi, Aaro; Ylikoski, Jukka; Mäkitie, Antti (2020)
    Objectives/Hypothesis We aimed to determine whether there was a difference between core and surface bacteriology of Finnish adults with recurrent or chronic tonsillitis to understand whether a surface swab is worthwhile and which bacteria are involved. Study Design Case series. Methods Uninflamed tonsillar surface swabs and core biopsies were taken prior to and during surgery, respectively, in 103 patients aged 16 to 66 years undergoing tonsillectomy for recurrent or chronic tonsillitis. The McNemar test was used to determine differences between the surface and core in the most prevalent bacterial species. Results Twenty-seven bacterial species were isolated in addition to normal flora and were more commonly found in the core (1.11 surface and 4.75 core bacteria isolated per patient). Viridans group streptococci were the most commonly detected bacteria, found in 88% of the patients, mainly in the core. The bacteria in general were mainly isolated from the core. Of the 10 most prevalent bacteria, only group C beta-hemolytic streptococci showed no difference between detection from core and surface swabs. Other bacteria found mainly in the core include Prevotella melaninogenica, Staphylococcus aureus, and fusobacteria. Conclusions There is discord between the surface and core bacteria. A different population of bacteria exists in the core, especially anaerobic bacteria, suggesting that a core sample may be useful in evaluating recurrent and chronic tonsillitis. Level of Evidence 4 Laryngoscope, 2019