Secondary metabolite from <em>Pseudomonas aeruginosa</em> LV strain exhibits antibacterial activity against <em>Staphylococcus aureus</em>
Metabólito secundário de Pseudomonas aeruginosa cepa LV exibe atividade antibacteriana em Staphylococcus aureus
DOI:
https://doi.org/10.34117/bjdv8n10-170Keywords:
antibiotic, antibiofilm, Fluopsin C, methicillin-sensitive Staphylococcus aureus, methicillin-resistant Staphylococcus aureusAbstract
This study aimed to evaluate the antibacterial activity of the dichloromethane/ethyl acetate fraction (named F4a), obtained from the culture of Pseudomonas aeruginosa LV strain in presence of copper chloride, against planktonic and sessile cells of Staphylococcus aureus, including those presenting multidrug resistance. First, the minimal inhibitory concentrations (MIC) of F4a for twenty-six clinical isolates were determined and the values ranged from 1.56 to 6.25 µg/mL. Minimal bactericidal concentration (MBC) of 3.13 µg/mL was detected in 84.6% of the isolates. The time-kill curve analysis revealed a significant decreased in colony-forming unit counts after 4 h of treatment with the MIC/MBC of F4a. Moreover, the MIC/MBC of the fluopsin C, a copper-containing compound present in F4a, were 1.56/3.13 µg/mL, indicating that this compound seems to be one of the active components related to the antibacterial activity against S. aureus. Images of transmission electron microscopy showed significant ultrastructural alterations in planktonic cells treated with the MIC/MBC of F4a. A significant reduction in the metabolic activity of established biofilms of all S. aureus isolates was observed after treatment with F4a. No hemolytic activity to human erythrocytes was detected for F4a, and the cytotoxic concentration to LLC-MK2 cells was 3.44 µg/mL. In conclusion, F4a exhibited a bactericidal activity against planktonic cells and inhibited the metabolic activity of biofilms of S. aureus. F4a can be promising for the development of new strategies for the treatment of infections caused by S. aureus.
References
Gatadi S, Gour J, Nanduri S. Natural Product Derived Promising Anti-MRSA Drug Leads: A Mini-Review. Bioorg Med Chem. 2019;27(17):3760-74.
Wu SC, Liu F, Zhu K, Shen JZ. Natural products that target virulence factors in antibiotic-resistant Staphylococcus aureus. J Agric Food Chem. 2019;67(48):13195-211.
Guo Y, Song G, Sun M, Wang J, Wang Y. Prevalence and Therapies of Antibiotic-Resistance in Staphylococcus aureus. Front Cell Infect Microbiol. 2020;10:107.
Vestergaard M, Frees D, Ingmer H. Antibiotic resistance and the MRSA problem. Microbiol Spectr. 2019;7(2).
Mehraj J, Witte W, Akmatov MK, Layer F, Werner G, Krause, G. Epidemiology of Staphylococcus aureus nasal carriage patterns in the community. How to Overcome the Antibiotic Crisis. Curr Top Microbiol Immunol. 2016;398:55-87.
Lister JL, Horswill AR. Staphylococcus aureus biofilms: recent developments in biofilm dispersal. Front Cell Infect Microbiol. 2014;4:178.
de Oliveira AG, Spago FR, Simionato AS, Navarro MO, da Silva CS, Barazetti AR, et al. Bioactive organocopper compound from Pseudomonas aeruginosa inhibits the growth of Xanthomonas citri subsp. citri. Front Microbiol. 2016;7:113.
Chopra S, Harjai K, Chhibber S. Antibiotic susceptibility of ica-positive and ica-negative MRSA in different phases of biofilm growth. J. Antibiot. 2015;68(1):15-22.
Molina-Manso D, del Prado G, Ortiz-Pérez A, Manrubia-Cobo M, Gómez-Barrena E, Cordero-Ampuero J, et al. In vitro susceptibility to antibiotics of staphylococci in biofilms isolated from orthopaedic infections. Int J Antimicrob Agents. 2013;41(6): 521-23.
Jones SM, Morgan M, Humphrey TJ, Lappin-Scott H. Effect of vancomycin and rifampicin on meticillin-resistant Staphylococcus aureus biofilms. Lancet. 2001; 357(9249):40-41.
Del Pozo JL. Biofilm-related disease. Expert Rev Anti Infect Ther. 2018;16(1):51-65.
Chang W, Ding D, Zhang S, Dai Y, Pan Q, Lu H, et al. Methicillin-resistant Staphylococcus aureus grown on vancomycin-supplemented screening agar displays enhanced biofilm formation. Antimicrob Agents Chemother. 2015;59(12):7906-10.
Mirani ZA, Jamil N. Effect of sub-lethal doses of vancomycin and oxacillin on biofilm formation by vancomycin intermediate resistant Staphylococcus aureus. J. Basic Microbiol. 2011;51(2):191-5.
Ortwine JK, Bhavan, K. Morbidity, mortality, and management of methicillin-resistant S. aureus bacteremia in the USA: update on antibacterial choices and understanding. Hosp Pract. 2018;46(2):64-72.
Zuniga A, Manalich J, Cortes R. Stethoscope or staphyloscope? Potential vector in nosocomial infections. Rev Chilena Infectol. 2016;33(1):19-25.
World Health Organization. Antibacterial agents in clinical development: an analysis of the antibacterial clinical development pipeline, including tuberculosis. No. WHO/EMP/IAU/2017.11. World Health Organization, 2017.
Rybtke M, Hultqvist LD, Givskov M, Tolker-Nielsen T. Pseudomonas aeruginosa biofilm infections: community structure, antimicrobial tolerance and immune response. J Mol Biol. 2015;427(23):3628-45.
Esposito E, Nastruzzi C, Sguizzato M, Cortesi R. Nanomedicines to Treat Skin Pathologies with Natural Molecules. Curr Pharm Des. 2019;25(21):2323-37.
Wright GD. Opportunities for natural products in 21st century antibiotic discovery. Nat Prod Rep. 2017;34(7):694-701.
Harvey A, Edrada-Ebel R, Quinn RJ. The re-emergence of natural products for drug discovery in the genomics era. Nat Rev Drug Discov. 2015;14(2):111-29.
Newman DJ, Cragg GM. Natural products as sources of new drugs over the 30 years from 1981 to 2010. J Nat Prod. 2012;75(3):311-35.
Butler MS, Blaskovich MA, Cooper MA. Antibiotics in the clinical pipeline in 2013. J Antibiot. 2013;66(10):571-91.
Clardy J, Fischbach MA, Walsh CT. New antibiotics from bacterial natural products. Nat Biotechnol. 2006;24(12):1541-50.
Walsh C. Antibiotics: actions, origins, resistance. American Society for Microbiology (ASM). 2003.
Craney A, Ahmed S, Nodwell J. Towards a new science of secondary metabolism. J Antibiot. 2013;66(7):387-400.
Buommino E, Scognamiglio M, Donnarumma G, Fiorentino A, D’Abrosca B. Recent advances in natural product-based anti-biofilm approaches to control infections. Mini-Rev Med Chem. 2014;14(14):1169-82.
Gross H, Loper JE. Genomics of secondary metabolite production by Pseudomonas spp. Nat Prod Rep. 2009;26(11):1408-46.
Andrade G. Process of production, purification and obtention of substances with antibiotic activity to control diseases caused by bacteria in plants. BR PI0803350-1A, 2008.
Pistori JF, Simionato AS, Navarro MOP, Andreata, MF, Santos IMO. Low-molecular-weight metabolites produced by Pseudomonas aeruginosa as an alternative to control Huanglongbing in Citrus sinensis cv. Valencia. Trop Plant Pathol. 2018;43(4):289-96.
Munhoz LD, Fonteque JP, Santos IMO, Navarro MOP, Simionato AS, Goya ET, et al. Control of bacterial stem rot on tomato by extracellular bioactive compounds produced by Pseudomonas aeruginosa LV strain. Cogent Food Agric. 2017;3:1282592.
Navarro MOP, Simionato AS, Pérez JCB, Barazetti AR, Emiliano J, Niekawa ETG, et al. Fluopsin C for treating multidrug-resistant infections: in vitro activity against clinically important strains and in vivo efficacy against carbapenemase-producing Klebsiella pneumoniae. Front Microbiol. 2019;10:2431.
Kerbauy G, Vivan ACP, Simões GC, Simionato AS, Pelisson M, Vespero EC, et al. Effect of a metalloantibiotic produced by Pseudomonas aeruginosa on Klebsiella pneumoniae Carbapenemase (KPC)-producing K. pneumoniae. Curr Pharm Biotechnol. 2016:17(4):389-97.
Cardozo VF, Oliveira AG, Nishio EK, Perugini MR, Andrade CG. Antibacterial activity of extracellular compounds produced by a Pseudomonas strain against methicillin-resistant Staphylococcus aureus (MRSA) strains. Ann Clin Microbiol Antimicrob. 2013;12(1):1-8.
Danelli T, Duarte FC, Oliveira TAD, Silva RSD, Frizon Alfieri D, Gonçalves GB, et al. Nasal Carriage by Staphylococcus aureus among Healthcare Workers and Students Attending a University Hospital in Southern Brazil: Prevalence, Phenotypic, and Molecular Characteristics. Interdiscip Perspect Infect Dis. 2020;2020:2020.
de Oliveira CF, Morey AT, Santos JP, Gomes LVP, Cardoso JD, Pinge-Filho P, et al. Molecular and phenotypic characteristics of methicillin-resistant Staphylococcus aureus isolated from hospitalized patients. J Infect Dev Ctries. 2015;9(7):743-51.
Amaral MM, Coelho LR, Flores RP, Souza RR, Silva-Carvalho MC, Teixeira LA, et al. The predominant variant of the Brazilian epidemic clonal complex of methicillin-resistant Staphylococcus aureus has an enhanced ability to produce biofilm and to adhere to and invade airway epithelial cells. J Infect Dis. 2005;192(5):801-10.
Bedoya JC, Dealis ML, Silva CS, Niekawa ETG, Navarro MOP, Simionato AS, et al. Enhanced production of target bioactive metabolites produced by Pseudomonas aeruginosa LV strain. Biocatal Agric Biotechnol. 2019;17:653-64.
CLSI. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard – Tenth Edition. CLSI Document M07-A11. Wayne, PA: Clinical and Laboratory Standards Institute; 2018.
Miles AA, Misra SS, Irwin JO. The estimation of the bactericidal power of the blood. J Hyg. 1938;38:732-49.
CLSI. Methods for Determining Bactericidal Activity of Antimicrobial Agents; Approved Guideline. CLSI Document M26-A. Wayne, PA: Clinical and Laboratory Standards Institute; 1999.
Stepanović S, Vuković D, Hola V, Bonaventura GD, Djukić S, Cirković I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. Apmis. 2007;115(8):891-99.
Schillaci D, Arizza V, Dayton T, Camarda L, Stefano VD. In vitro anti-biofilm activity of Boswellia spp. oleogum resin essential oils. Lett Appl Microbiol. 2008;47(5):433-38.
Walencka E, Różalska S, Sadowska B, Różalska B. The influence of Lactobacillus acidophilus-derived surfactants on staphylococcal adhesion and biofilm formation. Folia Microbiol. 2008;53(1):61-6.
Shi L, Ge HM, Tan SH, Li HQ, Song YC, Zhu HL, et al. Synthesis and antimicrobial activities of Schiff bases derived from 5-chloro-salicylaldehyde. Eur J Med Chem. 2007;42(4):558-64.
Teanpaisan R, Kawsud P, Pahumunto N, Puripattanavong J. Screening for antibacterial and antibiofilm activity in Thai medicinal plant extracts against oral microorganisms. J Tradit Complement Med. 2017;7(2):172-177.
Izumi E, Ueda-Nakamura T, Veiga Jr VF, Pinto AC, Nakamura CV. Terpenes from Copaifera demonstrated in vitro antiparasitic and synergic activity. J Med Chem. 2012;55(7):2994-3001.
Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect. 2012;18(3):268-81.
Zadrazilova I, Pospisilova S, Pauk K, Imramovsky A, Vinsova J, Cizek A, et al. In vitro bactericidal activity of 4-and 5-chloro-2-hydroxy-N-[1-oxo-1-(phenylamino) alkan-2-yl] benzamides against MRSA. Biomed Res Int. 2015;2015:349534.
Pankey GA, Sabath LD. Clinical relevance of bacteriostatic versus bactericidal mechanisms of action in the treatment of Gram-positive bacterial infections. Clin Infect Dis. 2004;38(6):864-70.
Schilcher K, Horswill AR. Staphylococcal biofilm development: structure, regulation, and treatment strategies. Microbiol Mol Biol Rev. 2020;84(3):e00026-19.
Patteson JB, Putz AT, Tao L, Simke WC, Bryant III LH, Britt RD, et al. Biosynthesis of fluopsin C, a copper-containing antibiotic from Pseudomonas aeruginosa. Science. 2021;374(6570):1005-9.
Navarro MOP, Dilarri G, Simionato AS, Grzegorczyk K, Dealis ML, Cano BG, et al. Determining the Targets of Fluopsin C Action on Gram-Negative and Gram-Positive Bacteria. Front Microbiol. 2020;11:1076.
Egawa Y, Umino K, Ito Y, Okuda T. Antibiotic YC 73 of Pseudomonas origin. II Structure and synthesis of thioformin and its cupric complex (YC 73). J Antibiot. 1971;24(2):124-30.
Itoh S, Inuzuka K, Suzuki T. New antibiotics produced by bacteria grown on n-paraffin (mixture of C12, C 13 and C14 fractions). J Antibiot. 1970;23:542–45.
Morita AKM, Ibelli-Bianco C, Anache JAA, Coutinho JV, Pelinson NS, Nobrega J, et al. Pollution threat to water and soil quality by dumpsites and non-sanitary landfills in Brazil: A review. Waste Manage. 2021;131:163-76.
Besold AN, Culbertson EM, Culotta VC. The Yin and Yang of copper during infection. J Biol Inorg Chem. 2016;21(2):137-44.
Del Rio LA, Gorgé JL, Olivares J, Mayor F. Antibiotics from Pseudomonas reptilivora II. Isolation, Purification, and Properties. Antimicrob Agents Chemother. 1972;2(3): 189-194.
Otsuka H, Niwayama S, Tanaka H, Take T, Uchiyama T. An antitumor antibiotic, no. 4601 from Streptomyces, identical with YC 73 of Pseudomonas origin. J. Antibiot. 1972;25:369–70.
