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DOUBLE DISK SYNERGY TEST AND BIOFILM FORMATION IN CLINICAL ISOLATES OF PSEUDOMONAS AERUGINOSA
DOUBLE DISK SYNERGY TEST AND BIOFILM FORMATION IN CLINICAL ISOLATES OF PSEUDOMONAS AERUGINOSA
Hozan Salahaddin Mahdi a and Sherko A Omer b
Hozan Salahaddin Mahdi a and Sherko A Omer b
a College of Science, University of Salahaddin.
b Department of Microbiology, College of Medicine, University of Sulaimani, Kurdistan Region, Iraq.
Submitted: 10/2/2019; Accepted: 29/7/2019; Published: 21/9/2019
DOI Link: https://doi.org/10.17656/jsmc.10207
ABSTRACT
Background
Pseudomonas aeruginosa is an opportunistic nosocomial pathogen responsible for several infections. For such infections, limited antibiotics are suggested and combination therapy and subsequent synergetic effects may be useful.
Objectives
To determine antimicrobial susceptibility and biofilm formation of clinical isolates of P. aeruginosa, and to test synergy between commonly used antimicrobials.
Patients and Methods
Pseudomonas aeruginosa isolates were collected from several hospitals and community health laboratories. The isolate's identities were confirmed, disk diffusion antimicrobial sensitivity test was performed and double-disk synergy test was carried out to detect synergism between seven antimicrobial combinations. The ability to form biofilm was tested using microtiter plate assay.
Results
One hundred clinical isolates of Pseudomonas aeruginosa were tested. Twenty-two isolates were from community laboratories, and 78 were from hospital laboratories. Thirty-four isolates were from urine, 32 from burn wound tissue, 13 from blood and 21 from other specimens. Polymyxin B was the most effective agent (92%) followed by meropenem (65%), while 75% of the isolates were resistant to ticarcillin-clavulanate and 59% to netilmicin. Forty synergism observations were detected between ticarcillin-clavulanate & netilmicin combination and 12 between ticarcillin-clavulanate & meropenem.
Conclusion
Ticarcillin-clavulanate was least effective while polymyxin B was more effective against clinical isolates of P. aeruginosa. Double-disk synergy revealed synergism with ticarcillin-clavulanate & netilmicin combination, disk synergy results can aid in deciding combination therapy. Biofilm formation was common in P. aeruginosa but was not found to affect disk synergy.
KEYWORDS
Pseudomonas aeruginosa, Antimicrobial Combination, Disk Synergy, Biofilm.
References
1. Stover CK, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P, Hickey MJ, et al. Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature. 2000;406(6799):959-64.
2. Weihui W, Yongxin J, Fang B, Shouguan J. Psedomonas aeruginosa. In: Yi-Wei Tang, Dongyou Liu, Joseph Schwartzman, Max Sussman, Ian Poxton, editors. Molecular Medical Microbiology. 2. Second ed: Academic Press; 2015. p. 753-67.
3. Hahn HP. The type-4 pilus is the major virulence-associated adhesin of Pseudomonas aeruginosa--a review. Gene. 1997;192(1):99-108.
4. Gellatly SL, Hancock RE. Pseudomonas aeruginosa: new insights into pathogenesis and host defenses. Pathog Dis. 2013;67(3):159-73.
5. Saleeb M, Sundin C, Aglar O, Pinto AF, Ebrahimi M, Forsberg A, et al. Structure-activity relationships for inhibitors of Pseudomonas aeruginosa exoenzyme S ADP-ribosyltransferase activity. Eur J Med Chem. 2018;143:568-76.
6. Liu PV. Extracellular toxins of Pseudomonas aeruginosa. The Journal of infectious diseases. 1974;130 Suppl(0):S94-9.
7. Szamosvari D, Reichle VF, Jureschi M, Bottcher T. Synthetic quinolone signal analogues inhibiting the virulence factor elastase of Pseudomonas aeruginosa. Chem Commun (Camb). 2016;52(92):13440-3.
8. Habash MB, Park AJ, Vis EC, Harris RJ, Khursigara CM. Synergy of silver nanoparticles and aztreonam against Pseudomonas aeruginosa PAO1 biofilms. Antimicrobial agents and chemotherapy. 2014;58(10):5818-30.
9. Lima J, Alves LR, Jacome P, Bezerra Neto JP, Maciel MAV, Morais MMC. Biofilm production by clinical isolates of Pseudomonas aeruginosa and structural changes in LasR protein of isolates non biofilm-producing. Braz J Infect Dis. 2018;22(2):129-36.
10. Forson OA, Ayanka E, Olu-Taiwo M, Pappoe-Ashong PJ, Ayeh-Kumi PJ. Bacterial infections in burn wound patients at a tertiary teaching hospital in Accra, Ghana. Ann Burns Fire Disasters. 2017;30(2):116-20.
11. Jung IY, Jeong SJ, Lee KM, Ahn JY, Ku NS, Han SH, et al. Risk factors for mortality in patients with Pseudomonas aeruginosa pneumonia: Clinical impact of mucA gene mutation. Respir Med. 2018;140:27-31.
12. Qi X, Qu H, Yang D, Zhou L, He YW, Yu Y, et al. Lower respiratory tract microbial composition was diversified in Pseudomonas aeruginosa ventilator-associated pneumonia patients. Respir Res. 2018;19(1):139.
13. Mittal R, Aggarwal S, Sharma S, Chhibber S, Harjai K. Urinary tract infections caused by Pseudomonas aeruginosa: A minireview. Journal of Infection and Public Health. 2009;2(3):101-11.
14. Kroin JS, Li J, Goldufsky JW, Gupta KH, Moghtaderi M, Buvanendran A, et al. Perioperative high inspired oxygen fraction therapy reduces surgical site infection with Pseudomonas aeruginosa in rats. J Med Microbiol. 2016;65(8):738-44.
15. Martin LW, Robson CL, Watts AM, Gray AR, Wainwright CE, Bell SC, et al. Expression of Pseudomonas aeruginosa antibiotic resistance genes varies greatly during infections in cystic fibrosis patients. Antimicrobial agents and chemotherapy. 2018.
16. Bassetti M, Vena A, Croxatto A, Righi E, Guery B. How to manage Pseudomonas aeruginosa infections. Drugs Context. 2018;7:212527.
17. Morita Y, Tomida J, Kawamura Y. Responses of Pseudomonas aeruginosa to antimicrobials. Frontiers in microbiology. 2014;4:422.
18. Kang C-I, Kim S-H, Kim H-B, Park S-W, Choe Y-J, Oh M-d, et al. Pseudomonas aeruginosa Bacteremia: Risk Factors for Mortality and Influence of Delayed Receipt of Effective Antimicrobial Therapy on Clinical Outcome. Clinical Infectious Diseases. 2003;37(6):745-51.
19. Chachanidze V, Curbelo-Irizarry A, Ashcraft D, Pankey G. In Vitro Synergy of Levofloxacin Plus Piperacillin/Tazobactam against Pseudomonas aeruginosa. Interdiscip Perspect Infect Dis. 2009;2009:984934.
20. Kobra Salimiyan Rizi KGaMKN. Adaptive Antibiotic Resistance: Overview and Perspectives. Journal of Infectious Diseases & Therapy. 2018;6(3).
21. Eliopoulos GM, Eliopoulos CT. Antibiotic combinations: should they be tested? Clin Microbiol Rev. 1988;1(2):139-56.
22. Traugott KA, Echevarria K, Maxwell P, Green K, Lewis JS, 2nd. Monotherapy or combination therapy? The Pseudomonas aeruginosa conundrum. Pharmacotherapy. 2011;31(6):598-608.
23. Tangden T. Combination antibiotic therapy for multidrug-resistant Gram-negative bacteria. Ups J Med Sci. 2014;119(2):149-53.
24. Biswas SM MM, Ara N , Ibrahim M , Nasir TA , Yunus S Comparison of Three Dimensional Test and Double Disc Synergy Test for detection of Extended Spectrum β-Lactamase (ESBL) producing Gram negative bacteria Pulse. 2013;6(1-2).
25. Marina V. Sukhorukova EYSMVENVI, Roman S. Kozlov. Evaluation of double-disk synergy test with low-content cephalosporin disks used by EUCAST versus high-content disks used by CLSI for detection of extended-spectrum beta-lactamases in Enterobacteriaceae. 26th European Congress of Clinical Microbiology and Infectious Diseases 9 - 12 April 2016; Amsterdam, Netherlands2016.
26. Punam Verma. Methods for Determining Bactericidal Activity and Antimicrobial Interactions: Synergy Testing, Time-Kill Curves, and Population Analysis. In: Richard Schwalbe, Lynn Steele-Moore, Avery C. Goodwin, editors. Antimicrobial Susceptibility Testing Protocols. Boca Raton CRC Press 2007. p. 275-98.
27. Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. American journal of clinical pathology. 1966;45(4):493-6.
28. CLSI. Performance standards for antimicrobial susceptibility testing. 28th ed. CLSI supplement M100S. Wayne, PA: Clinical and Laboratory Standards Institute; 2018.
29. Bodelon G, Montes-Garcia V, Lopez-Puente V, Hill EH, Hamon C, Sanz-Ortiz MN, et al. Detection and imaging of quorum sensing in Pseudomonas aeruginosa biofilm communities by surface-enhanced resonance Raman scattering. Nature materials. 2016;15(11):1203-11.
30. Saxena S, Banerjee G, Garg R, Singh M. Comparative Study of Biofilm Formation in Pseudomonas aeruginosa Isolates from Patients of Lower Respiratory Tract Infection. Journal of clinical and diagnostic research : JCDR. 2014;8(5):DC09-11.
31. Morrison JAJ, Wenzel RP. Epidemiology of Infections Due to Pseudomonas aeruginosa. Reviews of Infectious Diseases. 1984;6(Supplement_3):S627-S42.
32. Oliver A, Mulet X, Lopez-Causape C, Juan C. The increasing threat of Pseudomonas aeruginosa high-risk clones. Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy. 2015;21-22:41-59.
33. Yetkin G, Otlu B, Cicek A, Kuzucu C, Durmaz R. Clinical, microbiologic, and epidemiologic characteristics of Pseudomonas aeruginosa infections in a University Hospital, Malatya, Turkey. American journal of infection control. 2006;34(4):188-92.
34. OTHMAN, N., BABAKIR-MINA, M., NOORI, C. K. & RASHID, P. Y. 2014. Pseudomonas aeruginosa infection in burn patients in Sulaimaniyah, Iraq: risk factors and antibiotic resistance rates. J Infect Dev Ctries, 8, 1498-502.
35. Othman N, Babakir-Mina M, Noori CK, Rashid PY. Pseudomonas aeruginosa infection in burn patients in Sulaimaniyah, Iraq: risk factors and antibiotic resistance rates. Journal of infection in developing countries. 2014;8(11):1498-502.
36. Yadav V, Kiran V, Jaiswal M, Singh K. A study of antibiotic sensitivity pattern of Pseudomonas aeruginosa isolated from a tertiary care hospital in South Chhattisgarh. International Journal of Medical Science and Public Health. 2017;6(3):600-5.
37. Berditsch M, Jager T, Strempel N, Schwartz T, Overhage J, Ulrich AS. Synergistic effect of membrane-active peptides polymyxin B and gramicidin S on multidrug-resistant strains and biofilms of Pseudomonas aeruginosa. Antimicrobial agents and chemotherapy. 2015;59(9):5288-96.
38. Chatterjee M, Anju CP, Biswas L, Anil Kumar V, Gopi Mohan C, Biswas R. Antibiotic resistance in Pseudomonas aeruginosa and alternative therapeutic options. International journal of medical microbiology : IJMM. 2016;306(1):48-58.
39. Tre-Hardy M, Vanderbist F, Traore H, Devleeschouwer MJ. In vitro activity of antibiotic combinations against Pseudomonas aeruginosa biofilm and planktonic cultures. International journal of antimicrobial agents. 2008;31(4):329-36.
40. Shih P-C, Huang C-T. Effects of quorum-sensing deficiency on Pseudomonas aeruginosa biofilm formation and antibiotic resistance. Journal of Antimicrobial Chemotherapy. 2002;49(2):309-14.
41. Fahmy A, Srinivasan A, Webber MA. The Relationship Between Bacterial Multidrug Efflux Pumps and Biofilm Formation. In: Li X-Z, Elkins CA, Zgurskaya HI, editors. Efflux-Mediated Antimicrobial Resistance in Bacteria: Mechanisms, Regulation and Clinical Implications. Cham: Springer International Publishing; 2016. p. 651-63.
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