Antimicrobial and Antibiofilm Effect of 4,4'-Dihydroxy-azobenzene Against Clinically Resistant Staphylococci

Overview

Journal Antibiotics (Basel)

Specialty Pharmacology

Date 2022 Dec 23

PMID 36551456

Authors

Maria Perez-Aranda

Eloisa Pajuelo

Salvadora Navarro-Torre

Patricia Perez-Palacios

Belen Begines

Ignacio D Rodriguez-Llorente

Yadir Torres

Ana Alcudia

Affiliations

Soon will be listed here.

Abstract

The spread of antibiotic resistance among human and animal pathogens is one of the more significant public health concerns. Moreover, the restrictions on the use of particular antibiotics can limit the options for the treatment of infections in veterinary clinical practice. In this context, searching for alternative antimicrobial substances is crucial nowadays. In this study, 4,4'-dihydroxy-azobenzene (DHAB) was tested for its potential in vitro as an antimicrobial agent against two relevant human and animal pathogens, namely and . The values of minimal inhibitory concentration (MIC) were 64 and 32 mg/L respectively, and they comparable to other azo compounds of probed antimicrobial activity. In addition, the minimal bactericidal concentrations (MCB) were 256 and 64 mg/L. The mechanism by which DHAB produces toxicity in staphylococci has been investigated. DHAB caused membrane damage as revealed by the increase in thiobarbituric acid reactive substances (TBARS) such as malondialdehyde. Furthermore, differential induction of the enzymes peroxidases and superoxide dismutase in and suggested their prevalent role in ROS-scavenging due to the oxidative burst induced by this compound in either species. In addition, this substance was able to inhibit the formation of biofilms by both bacteria as observed by colorimetric tests and scanning electron microscopy. In order to assess the relevance of DHAB against clinical strains of MRSA, 10 clinical isolates resistant to either methicillin or daptomycin were assayed; 80% of them gave values of CMI and CMB similar to those of the control strain. Finally, cutaneous plasters containing a composite formed by an agar base supplemented with DHAB were designed. These plasters were able to inhibit in vitro the growth of and , particularly the later, and this suggests that this substance could be a promising candidate as an alternative to antibiotics in the treatment of animal skin infections, as it has been proven that the toxicity of this substance is very low particularly at a dermal level.

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References

Myers J, Hennessey M, Arnold J, McCubbin K, Lembo T, Mateus A . Crossover-Use of Human Antibiotics in Livestock in Agricultural Communities: A Qualitative Cross-Country Comparison between Uganda, Tanzania and India. Antibiotics (Basel). 2022; 11(10). PMC: 9598773. DOI: 10.3390/antibiotics11101342. View

Bedekar P, Saratale R, Saratale G, Govindwar S . Oxidative stress response in dye degrading bacterium Lysinibacillus sp. RGS exposed to Reactive Orange 16, degradation of RO16 and evaluation of toxicity. Environ Sci Pollut Res Int. 2014; 21(18):11075-85. DOI: 10.1007/s11356-014-3041-2. View

Cattelan N, Jennings-Gee J, Dubey P, Yantorno O, Deora R . Hyperbiofilm Formation by Bordetella pertussis Strains Correlates with Enhanced Virulence Traits. Infect Immun. 2017; 85(12). PMC: 5695122. DOI: 10.1128/IAI.00373-17. View

Pan H, Feng J, He G, Cerniglia C, Chen H . Evaluation of impact of exposure of Sudan azo dyes and their metabolites on human intestinal bacteria. Anaerobe. 2012; 18(4):445-53. PMC: 5870115. DOI: 10.1016/j.anaerobe.2012.05.002. View

Chakraborty S, Mahapatra S, Roy S . Biochemical characters and antibiotic susceptibility of Staphylococcus aureus isolates. Asian Pac J Trop Biomed. 2013; 1(3):212-6. PMC: 3609185. DOI: 10.1016/S2221-1691(11)60029-4. View

Pan H, Feng J, Cerniglia C, Chen H . Effects of Orange II and Sudan III azo dyes and their metabolites on Staphylococcus aureus. J Ind Microbiol Biotechnol. 2011; 38(10):1729-38. PMC: 5868333. DOI: 10.1007/s10295-011-0962-3. View

Tenover F, Arbeit R, Goering R, Mickelsen P, Murray B, Persing D . Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995; 33(9):2233-9. PMC: 228385. DOI: 10.1128/jcm.33.9.2233-2239.1995. View

Cox Jr L, Ricci P . Causal regulations vs. political will: why human zoonotic infections increase despite precautionary bans on animal antibiotics. Environ Int. 2008; 34(4):459-75. DOI: 10.1016/j.envint.2007.10.010. View

Andersson D, Balaban N, Baquero F, Courvalin P, Glaser P, Gophna U . Antibiotic resistance: turning evolutionary principles into clinical reality. FEMS Microbiol Rev. 2020; 44(2):171-188. DOI: 10.1093/femsre/fuaa001. View

10.

Vollaro A, Esposito A, Antonaki E, Iula V, DAlonzo D, Guaragna A . Steroid Derivatives as Potential Antimicrobial Agents Against Planktonic Cells. Microorganisms. 2020; 8(4). PMC: 7232480. DOI: 10.3390/microorganisms8040468. View

11.

Ben Mansour H, Corroler D, Barillier D, Ghedira K, Chekir L, Mosrati R . Evaluation of genotoxicity and pro-oxidant effect of the azo dyes: acids yellow 17, violet 7 and orange 52, and of their degradation products by Pseudomonas putida mt-2. Food Chem Toxicol. 2007; 45(9):1670-7. DOI: 10.1016/j.fct.2007.02.033. View

12.

MAEHLY A, Chance B . The assay of catalases and peroxidases. Methods Biochem Anal. 1954; 1:357-424. DOI: 10.1002/9780470110171.ch14. View

13.

Somayaji R, Priyantha M, Rubin J, Church D . Human infections due to Staphylococcus pseudintermedius, an emerging zoonosis of canine origin: report of 24 cases. Diagn Microbiol Infect Dis. 2016; 85(4):471-6. DOI: 10.1016/j.diagmicrobio.2016.05.008. View

14.

Sabino H, Valera F, Santos D, Fantucci M, Titoneli C, Martinez R . Biofilm and Planktonic Antibiotic Resistance in Patients With Acute Exacerbation of Chronic Rhinosinusitis. Front Cell Infect Microbiol. 2022; 11:813076. PMC: 8801706. DOI: 10.3389/fcimb.2021.813076. View

15.

Ghosh C, Sarkar P, Issa R, Haldar J . Alternatives to Conventional Antibiotics in the Era of Antimicrobial Resistance. Trends Microbiol. 2019; 27(4):323-338. DOI: 10.1016/j.tim.2018.12.010. View

16.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

17.

Concilio S, Sessa L, Petrone A, Porta A, Diana R, Iannelli P . Structure Modification of an Active Azo-Compound as a Route to New Antimicrobial Compounds. Molecules. 2017; 22(6). PMC: 6152751. DOI: 10.3390/molecules22060875. View

18.

Kaur S, Kaur A . Variability in antioxidant/detoxification enzymes of Labeo rohita exposed to an azo dye, acid black (AB). Comp Biochem Physiol C Toxicol Pharmacol. 2014; 167:108-16. DOI: 10.1016/j.cbpc.2014.06.009. View

19.

Abdullahi I, Fernandez-Fernandez R, Juarez-Fernandez G, Martinez-Alvarez S, Eguizabal P, Zarazaga M . Wild Animals Are Reservoirs and Sentinels of and MRSA Clones: A Problem with "One Health" Concern. Antibiotics (Basel). 2021; 10(12). PMC: 8698730. DOI: 10.3390/antibiotics10121556. View

20.

Beyer Jr W, Fridovich I . Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal Biochem. 1987; 161(2):559-66. DOI: 10.1016/0003-2697(87)90489-1. View