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molecules Article Biofilm Eradication Using Biogenic Silver Nanoparticles María Belén Estevez 1, Sofía Raffaelli 1, Scott G. Mitchell 2,3,* , Ricardo Faccio 4 and Silvana Alborés 1,* 1 2 3 4 * Correspondence: scott@unizar.es (S.G.M.); salbores@fq.edu.uy (S.A.) Academic Editor: Russell Crawford Received: 28 March 2020; Accepted: 24 April 2020; Published: 26 April 2020 Área de Microbiología, Departamento de Biociencias, Facultad de Química, Universidad de la República, Montevideo 11800, Uruguay; bestevez@fq.edu.uy (M.B.E.); sraffaelli@fq.edu.uy (S.R.) Instituto de Ciencia de Materiales de Aragón (ICMA), Consejo Superior de Investigaciones Científicas (CSIC)-Universidad de Zaragoza, 50009 Zaragoza, Spain CIBER-BBN, Instituto de Salud Carlos III, 28029 Madrid, Spain Centro NanoMat & Grupo Física, Departamento de Experimentación y Teoría de la Estructura de la Materia y sus Aplicaciones (DETEMA), Facultad de Química, Universidad de la República, Montevideo 11800, Uruguay; rfaccio@fq.edu.uy Abstract: Microorganisms offer an alternative green and scalable technology for the synthesis of value added products. Fungi secrete high quantities of bioactive substances, which play dual-functional roles as both reducing and stabilizing agents in the synthesis of colloidal metal nanoparticles such as silver nanoparticles, which display potent antimicrobial properties that can be harnessed for a number of industrial applications. The aim of this work was the production of silver nanoparticles using the extracellular cell free extracts of Phanerochaete chrysosporium, and to evaluate their activity as antimicrobial and antibiofilm agents. The 45–nm diameter silver nanoparticles synthesized using this methodology possessed a high negative surface charge close to −30 mV and showed colloidal stability from pH 3–9 and under conditions of high ionic strength ([NaCl] = 10–500 mM). A combination of environmental SEM, TEM, and confocal Raman microscopy was used to study the nanoparticle-E. coli interactions to gain a first insight into their antimicrobial mechanisms. Raman data demonstrate a significant decrease in the fatty acid content of E. coli cells, which suggests a loss of the cell membrane integrity after exposure to the PchNPs, which is also commensurate with ESEM and TEM images. Additionally, these biogenic PchNPs displayed biofilm disruption activity for the eradication of E. coli and C. albicans biofilms. Keywords: biogenic nanoparticles; silver nanoparticles; antimicrobial; antibiofilm 1. Introduction Biological synthesis is an alternative green technology used for the production of metal nanoparticles, compared to the conventional physical and chemical methods that may use toxic and/or expensive reagents and severe reaction conditions [1]. Some microorganisms are able to accumulate and detoxify heavy metals due to various reductase enzymes, which reduce metal salts to metal nanoparticles with a narrow size distribution. For this reason, microorganisms have immense potential as ecofriendly and cost-effective tools for biological synthesis of nanoparticles, avoiding toxic, harsh chemicals and the high energy demand required for physiochemical synthesis [2] Biological synthesis by microorganisms includes the intracellular or extracellular production [3,4]. Extracellular synthesis of nanoparticles has received much attention because it eliminates the downstream processing steps required for the recovery of nanoparticles in intracellular methodologies, such as sonication to break Molecules 2020, 25, 2023; doi:10.3390/molecules25092023 www.mdpi.com/journal/moleculesPDF Image | Biofilm Eradication Using Biogenic Silver Nanoparticles
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