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Effect of Biosynthesized Zinc oxide Nanoparticles on Phenotypic and Genotypic Biofilm Formation of Proteus mirabilis




Biofilm, Green synthesis, LuxS, Nanoparticles, Zinc oxide


Proteus mirabilis is considered as a third common cause of catheter-associated urinary tract infection, with urease production, the potency of catheter blockage due to the formation of biofilm formation is significantly enhanced. Biofilms are major virulence factors expressed by pathogenic bacteria to resist antibiotics; in this concern the need for providing new alternatives for antibiotics is getting urgent need, This study aimed to explore whether green synthesized zinc oxide nanoparticles (ZnO NPs) can function as an anti-biofilm agent produced by P.mirabilis. Bacterial cells were capable of catalyzing the biosynthesis process by producing reductive enzymes. The nanoparticles were synthesized from cell free extract of P.mirabilis. Characterization of biosynthesized zinc nanoparticles was carried out to determine the chemical and physical properties of the product using AFM, TEM, FESEM, XRD and UV visible spectrometry. The hexagonal structure was confirmed by XRD, Particle size was marked at 84.45 nm by TEM, FESEM was used to confirm the surface morphology. AFM analysis was used to reveal the roughness and distribution of nanoparticles. UV–visible spectra of the synthesized nanoparticles recorded maximum peak at 287 nm. Zinc nanoparticles showed remarkable biofilm inhibitory effect on clinical isolates of multidrug resistant Proteus mirabilis. Strong biofilm producer strains show weak biofilm production After incubation for 24 and 48 hours at 37Co with 32 μg/ml sub -MIC concentration of ZnO nanoparticles. Down regulation changes in LuxS expression using Real time PCR technology were detected after treatment with zink nanoparticles of these isolates compared to untreated isolates. From all findings conducted by this study, zinc oxide nanoparticles can function as anti-bacterial agent in concentration dependent manner.


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Muhammad MH, Idris AL, Fan X, Guo Y, Yu Y, Jin X, et al. Beyond risk: Bacterial biofilms and their regulating approaches. Front Microbiol. 2020; 11: 928.

Shokouhfard M, Kermanshahi R-K, Feizabadi M-M, Teimourian S, Safari F. Lactobacillus spp. derived biosurfactants effect on expression of genes involved in Proteus mirabilis biofilm formation. Infect Genet Evol . 2022; 100(105264): 105264.

Spirescu VA, Șuhan R, Niculescu AG, Grumezescu V, Negut I, Holban AM, et al. Biofilm-resistant nanocoatings based on zno nanoparticles and linalool. Nanomaterials. 2021; 11(10): 2564.

Balaure PC, Grumezescu AM. Recent advances in surface nanoengineering for biofilm prevention and control. Part ii: active, combined active and passive, and smart bacteria-responsive antibiofilm nanocoatings. Nanomaterials. 2020; 10(8): 1527.

Kazemzadeh-Narbat M, Cheng H, Chabok R, Alvarez MM, de la Fuente-Nunez C, Phillips KS, et al. Strategies for antimicrobial peptide coatings on medical devices: a review and regulatory science perspective. Crit Rev Biotechnol. 2021; 41(1): 94–120.

Alwash A. The green synthesize of zinc oxide catalyst using pomegranate peels extract for the photocatalytic degradation of methylene blue dye. Baghdad Sci J. 2020; 17(3): 0787–0787.

Kadhim AA, Salman JAS, Haider AJ, Ibraheem SA, Kadhim H ali. Effect of zinc oxide nanoparticles biosynthesized by leuconostoc mesenteroides ssp. Dextranicum against bacterial skin infections. In: 2019 12th (DeSE): IEEE; 2019: 755–760.

Jiang Q, Chen J, Yang C, Yin Y, Yao K. Quorum sensing: a prospective therapeutic target for bacterial diseases. Biomed Res Int. 2019; 2019: 1–15.

Wiegand I, Hilpert K, Hancock REW. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc. 2008; 3(2): 163–75.

Banerjee S, Vishakha K, Das S, Dutta M, Mukherjee D, Mondal J, et al. Antibacterial, anti-biofilm activity and mechanism of action of pancreatin doped zinc oxide nanoparticles against methicillin resistant Staphylococcus aureus. Colloids Surf B Biointerfaces. 2020; 190: 110921.

Husain FM, Khan MS, Ahmad I, Khan RA, Al-Shabib NA, Oves M, et al. Nanomaterials as a novel class of anti-infective agents that attenuate bacterial quorum sensing. In: Ahmad I, Ahmad S, Rumbaugh KP (eds.) Antibacterial Drug Discovery to Combat MDR. Springer Singapore; 2019. p. 581–604.

Ali MdA, Ahmed T, Wu W, Hossain A, Hafeez R, Islam Masum MdM, et al. Advancements in plant and microbe-based synthesis of metallic nanoparticles and their antimicrobial activity against plant pathogens. Nanomaterials. 2020; 10(6): 1146.

Phang YK, Aminuzzaman M, Akhtaruzzaman Md, Muhammad G, Ogawa S, Watanabe A, et al. Green synthesis and characterization of cuo nanoparticles derived from papaya peel extract for the photocatalytic degradation of palm oil mill effluent(Pome). Sustainability. 2021; 13(2): 796.

Jaffar N, Miyazaki T, Maeda T. Biofilm formation of periodontal pathogens on hydroxyapatite surfaces: Implications for periodontium damage: Biofilm Formation of Periodontal Pathogens on Hydroxyapatite Surfaces. J Biomed Mater Res A . 2016; 104(11): 2873–2880.

Gajdács M, Urbán E. Comparative epidemiology and resistance trends of proteae in urinary tract infections of inpatients and outpatients: a 10-year retrospective study. Antibiotics. 2019; 8(3): 91.

Punniyakotti P, Panneerselvam P, Perumal D, Aruliah R, Angaiah S. Anti-bacterial and anti-biofilm properties of green synthesized copper nanoparticles from Cardiospermum halicacabum leaf extract. Bioprocess Biosyst Eng. 2020; 43(9): 1649–1657.

Passat DNF. Local Study of blaCTX-M genes detection in Proteus spp. by using PCR technique. Iraqi J Sci. 2016; 57(2c): 1371–1376.

Kang Q, Wang X, Zhao J, Liu Z, Ji F, Chang H, et al. Multidrug‐resistant Proteus mirabilis isolates carrying bla OXA‐1 and bla NDM‐1 from wildlife in China: increasing public health risk. Integr Zool. 2021; 16(6): 798–809.

Little K, Austerman J, Zheng J, Gibbs KA. Cell shape and population migration are distinct steps of Proteus mirabilis swarming that are decoupled on high-percentage agar. J Bacteriol. 2019; 201(11): e00726-18.

Tuson HH, Copeland MF, Carey S, Sacotte R, Weibel DB. Flagellum density regulates Proteus mirabilis swarmer cell motility in viscous environments. J Bacteriol. 2013; 195(2): 368–77.

Grasso G, Zane D, Dragone R. Microbial nanotechnology: challenges and prospects for green biocatalytic synthesis of nanoscale materials for sensoristic and biomedical applications. Nanomaterials. 2019; 10(1): 11.

Kouhkan M, Ahangar P, Babaganjeh LA, Allahyari-Devin M. Biosynthesis of copper oxide nanoparticles using lactobacillus casei subsp. Casei and its anticancer and antibacterial activities. Curr Nanosci. 2020; 16(1): 101–111.

Janani B, Syed A, Raju LL, Al-Harthi HF, Thomas AM, Das A, et al. Synthesis of carbon stabilized zinc oxide nanoparticles and evaluation of its photocatalytic, antibacterial and anti-biofilm activities. J Inorg Organomet Polym Mater. 2020; 30(6): 2279–2288.

Duffy LL, Osmond-McLeod MJ, Judy J, King T. Investigation into the antibacterial activity of silver, zinc oxide and copper oxide nanoparticles against poultry-relevant isolates of Salmonella and Campylobacter. Food Control. 2018; 92: 293–300.

AL-Asady ZM, AL-Hamdani AH, Hussein MA. Study the optical and morphology properties of zinc oxide nanoparticles. In: Baghdad, Iraq; AIP Conf Proc. 2020. 2213(1) p. 020061.

Kassinger SJ, van Hoek ML. Biofilm architecture: An emerging synthetic biology target. Synth Syst Biotechnol .2020; 5(1): 1–10.

Khan MohdF, Husain FM, Zia Q, Ahmad E, Jamal A, Alaidarous M, et al. Anti-quorum sensing and anti-biofilm activity of zinc oxide nanospikes. ACS Omega. 2020; 5(50): 32203–32215.

Siddiqi KS, ur Rahman A, Tajuddin, Husen A. Properties of zinc oxide nanoparticles and their activity against microbes. Nanoscale Res Lett. 2018; 13(1): 141.

Shah S, Gaikwad S, Nagar S, Kulshrestha S, Vaidya V, Nawani N, et al. Biofilm inhibition and anti-quorum sensing activity of phytosynthesized silver nanoparticles against the nosocomial pathogen Pseudomonas aeruginosa. Biofouling .2019; 35(1): 34 49.:

Abadeer NS, Fülöp G, Chen S, Käll M, Murphy CJ. Interactions of bacterial lipopolysaccharides with gold nanorod surfaces investigated by refractometric sensing. ACS Appl Mater Interfaces .2015; 7(44): 24915–25.

Hasson SO, kadhem Salman SA, Hassan SF, Abbas SM. Antimicrobial effect of eco- friendly silver nanoparticles synthesis by iraqi date palm (Phoenix dactylifera) on gram-negative biofilm-forming bacteria. Baghdad Sci J. 2021; 18(4): 1149-1156.

Bondarenko OM, Sihtmäe M, Kuzmičiova J, Ragelienė L, Kahru A, Daugelavičius R. Plasma membrane is the target of rapid antibacterial action of silver nanoparticles in Escherichia coli and Pseudomonas aeruginosa. Int J Nanomedicine. 2018; 13: 6779–6790.

Hussein EI, Al-Batayneh K, Masadeh MM, Dahadhah FW, Al Zoubi MS, Aljabali AA, et al. Assessment of pathogenic potential, virulent genes profile, and antibiotic susceptibility of proteus mirabilis from urinary tract infection. Int J Microbiol. 2020; 2020: 1–5.

Rajalakshmi, Sangeetha, Udhaya. Effect of antifungal drugs against candida isolates from diabetic women with vaginitis. J Infect Dis Ther. 2017; 05(04): 1-5.

Jabin Mishu N, Sm S, Hm K, Nabonee MA, zannat dola N, haque A. Association between biofilm formation and virulence genes expression and antibiotic resistance pattern in proteus mirabilis, isolated from patients of dhaka medical college hospital. Arch Clin Biomed Res. 2022; 06(03): 418-434.

Badawy MSEM, Riad OKM, Taher FA, Zaki SA. Chitosan and chitosan-zinc oxide nanocomposite inhibit expression of LasI and RhlI genes and quorum sensing dependent virulence factors of Pseudomonas aeruginosa. Int J Biol Macromol. 2020; 149: 1109–1117.

Abdul-Hamza HK, Mohammed GJ. Anti-quorum sensing effect of streptococcus agalatiaceae by Zinc Oxide, Copper Oxide, and Titanium Oxide nanoparticles. J Phys : Conf Ser . 2021; 1999(1): 012031.

Cai X, Liu X, Jiang J, Gao M, Wang W, Zheng H, et al. Molecular mechanisms, characterization methods, and utilities of nanoparticle biotransformation in nanosafety assessments. Small. 2020;1 6(36): 1907663.

Joshi ASPAMI. nteractions of gold and silver nanoparticles with bacterial biofilms: Molecular interactions behind inhibition and resistance. Int J Mol Sci.. 2020; 21(20).

Sistemática D, Gabriela SS, Daniela FR, Helia BT. Zinc nanoparticles as potential antimicrobial agent in disinfecting root canals. Odontostomat. 2016; 10(3): 547–54.

Peulen T-O, Wilkinson KJ. Diffusion of nanoparticles in a biofilm. Environ Sci Technol. 2011; 45(8): 3367–73.

Raheem H, Yasser H. Silver Nanoparticles as Antibacterial Action against Pseudomonas Fluorescens Isolated from Burn Infection. Ann Romanian Soc Cell Biol. 2021; 25(4): 12578–83.

Chaudhary A, Kumar N, Kumar R, Salar RK. Antimicrobial activity of zinc oxide nanoparticles synthesized from Aloe vera peel extract. SN Appl Sci . 2019; 1: 136.

Gómez-Gómez B, Arregui L, Serrano S, Santos A, Pérez-Corona T, Madrid Y. Unravelling mechanisms of bacterial quorum sensing disruption by metal-based nanoparticles. Sci Total Environ. 2019; 696: 133869.

Alavi M, Li L, Nokhodchi A. Metal, metal oxide and polymeric nanoformulations for the inhibition of bacterial quorum sensing. Drug Discov Today. 2022; 28(1): 103392.

Rashid AE, Ahmed ME, Hamid MK. Evaluation of Antibacterial and Cytotoxicity Properties of Zinc Oxide Nanoparticles Synthesized by Precipitation Method against Methicillin-resistant Staphylococcus aureus. Int J Drug Deliv Technol. 2022; 12(3): 985-989.