Abstract
The study evaluated and compared the effect of adding streptokinase and amylase to antibiotics that are already used in clinical practice to treat Gram negative bacteria biofilm infection on indwelling devices on the antibiotics' minimum inhibitory concentration (MIC). 24 h-old biofilms were developed on 96-well plate with eight clinical isolates. MIC of amikacin, cefepime, ceftazidime, colistin, meropenem, and piperacillin–tazobactam, on biofilms were measured before and after the addition of 25 U/ml streptokinase and 25 μg/ml amylase with microplate reader. The addition of streptokinase reduces the MICs of cefepime, ceftazidime, colistin, meropenem from (16, 16, 8, 4 μg/ml) to (8, 1, 1, 0.5 μg/ml) in Escherichia coli (isolate 1). While the addition of amylase reduces the MICs of only cefepime, ceftazidime from (16, 16 μg/ml) to (2, 4 μg/ml) in E. coli (isolate 1). In Pseudomonas aeruginosa (isolate 4), the MICs of amikacin, cefepime, ceftazidime, colistin and meropenem (64, 16, 32, 4, 32 μg/ml) reduced to (2, 1, 0.5, 0.25, 0.5 μg/ml) with streptokinase and (4, 4, 4, 2, 0.5 μg/ml) with amylase respectively. Similar inhibitions were seen in Pseudomonas putida, Proteus mirabilis. We can conclude that the addition of streptokinase and amylase were effective in reducing the MICs of antibiotics that are commonly used to treat Gram negative bacteria biofilm infection on indwelling devices, thereby increasing susceptibility of bacteria to antibiotics. Streptokinase obviously had a greater effect than amylase, implying that it should be prioritized in future in vivo and clinical studies to obtain successful therapy with antibiotics on biofilm infections.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
O’Toole G, Kaplan HB, Kolter R (2000) Biofilm formation as microbial development. Annu Rev Microbiol 54:49–79. https://doi.org/10.1146/annurev.micro.54.1.49
Hall-Stoodley L, Stoodley P (2009) Evolving concepts in biofilm infections. Cell Microbiol 11:1034–1043. https://doi.org/10.1111/j.1462-5822.2009.01323.x
Jamal M, Ahmad W, Andleeb S et al (2018) Bacterial biofilm and associated infections. J Chin Med Assoc 81:7–11. https://doi.org/10.1016/j.jcma.2017.07.012
Kokare CR, Chakraborty S, Khopade AN, Mahadik KR (2009) Biofilm: importance and applications. Indian J Biotechnol 6:159–168
Perciva S, Williams D, Cooper T, Randle J (2014) Biofilms in infection prevention and control a healthcare handbook. Academic Press, Cambridge, Massachusetts
Tran PL, Lowry N, Campbell T et al (2012) An organoselenium compound inhibits Staphylococcus aureus biofilms on hemodialysis catheters in vivo. Antimicrob Agents Chemother 56:972–978. https://doi.org/10.1128/AAC.05680-11
Høiby N, Bjarnsholt T, Givskov M et al (2010) Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents 35:322–332. https://doi.org/10.1016/j.ijantimicag.2009.12.011
Hengzhuang W, Wu H, Ciofu O et al (2011) Pharmacokinetics/pharmacodynamics of colistin and imipenem on mucoid and nonmucoid Pseudomonas aeruginosa biofilms. Antimicrob Agents Chemother 55:4469–4474. https://doi.org/10.1128/AAC.00126-11
Roy R, Tiwari M, Donelli G, Tiwari V (2018) Strategies for combating bacterial biofilms: a focus on anti-biofilm agents and their mechanisms of action. Virulence 9:522–554. https://doi.org/10.1080/21505594.2017.1313372
Pieper-Bigelow C, Strocchi A, Levitt MD (1990) Where does serum amylase come from and where does it go? Gastroenterol Clin North Am 19:793–810
Azzopardi E, Lloyd C, Teixeira SR et al (2016) Clinical applications of amylase: novel perspectives. Surgery 160:26–37. https://doi.org/10.1016/j.surg.2016.01.005
Schachtele CF, Staat RH, Harlander SK (1975) Dextranases from oral bacteria: inhibition of water-insoluble glucan production and adherence to smooth surfaces by Streptococcus mutans. Infect Immun 12:309–317. https://doi.org/10.1128/iai.12.2.309-317.1975
Lembre P, Lorentz C, Di P (2012) Exopolysaccharides of the biofilm matrix: a complex biophysical world. In: Karunaratne DN (Ed) The complex world of polysaccharides. InTechOpen, London, pp 371–392. https://doi.org/10.5772/51213
Kalpana BJ, Aarthy S, Pandian SK (2012) Antibiofilm activity of α-amylase from Bacillus subtilis S8–18 against biofilm forming human bacterial pathogens. Appl Biochem Biotechnol 167:1778–1794. https://doi.org/10.1007/s12010-011-9526-2
Jee S-C, Kim M, Sung J-S, Kadam AA (2020) Efficient biofilms eradication by enzymatic-cocktail of pancreatic protease type-I and bacterial α-amylase. Polymers (Basel) 12:3032. https://doi.org/10.3390/polym12123032
Solihin J, Waturangi DE, Purwadaria T (2021) Induction of amylase and protease as antibiofilm agents by starch, casein, and yeast extract in Arthrobacter sp. CW01. BMC Microbiol 21:232. https://doi.org/10.1186/s12866-021-02294-z
Nemoto K, Hirota K, Ono T et al (2000) Effect of varidase (streptokinase) on biofilm formed by Staphylococcus aureus. Chemotherapy 46:111–115. https://doi.org/10.1159/000007264
Vanassche T, Peetermans M, Van Aelst LNL et al (2013) The role of staphylothrombin-mediated fibrin deposition in catheter-related Staphylococcus aureus infections. J Infect Dis 208:92–100. https://doi.org/10.1093/infdis/jit130
Kwiecinski J, Peetermans M, Liesenborghs L et al (2016) Staphylokinase control of Staphylococcus aureus biofilm formation and detachment through host plasminogen activation. J Infect Dis 213:139–148. https://doi.org/10.1093/infdis/jiv360
Zapotoczna M, McCarthy H, Rudkin JK et al (2015) An essential role for coagulase in Staphylococcus aureus biofilm development reveals new therapeutic possibilities for device-related infections. J Infect Dis 212:1883–1893. https://doi.org/10.1093/infdis/jiv319
Baldassarri L, Creti R, Recchia S et al (2006) Therapeutic failures of antibiotics used to treat macrolide-susceptible Streptococcus pyogenes infections may be due to biofilm formation. J Clin Microbiol 44:2721–2727. https://doi.org/10.1128/JCM.00512-06
Antunes ALS, Trentın DS, Bonfanti JW et al (2010) Application of a feasible method for determination of biofilm antimicrobial susceptibility in Staphylococci. APMIS 118:873–877. https://doi.org/10.1111/j.1600-0463.2010.02681.x
Aksoy N, Vatansever C, Zengin Ersoy G et al (2022) The effect of biofilm inhibitor N-acetylcysteine on the minimum inhibitory concentration of antibiotics used in Gram-negative bacteria in the biofilm developed on catheters. Int J Artif Organs 45:865–870. https://doi.org/10.1177/03913988221112969
Stamm WE (1978) Infections related to medical devices. Ann Intern Med 89:764. https://doi.org/10.7326/0003-4819-89-5-764
Donlan RM, Costerton JW (2002) Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15:167–193. https://doi.org/10.1128/CMR.15.2.167-193.2002
Lewis K (2007) Persister cells, dormancy and infectious disease. Nat Rev Microbiol 5:48–56. https://doi.org/10.1038/nrmicro1557
Lewis K (2001) Riddle of biofilm resistance. Antimicrob Agents Chemother 45:999–1007. https://doi.org/10.1128/AAC.45.4.999-1007.2001
Jørgensen N, Zobek N, Dreier C et al (2016) Streptokinase treatment reverses biofilm-associated antibiotic resistance in Staphylococcus aureus. Microorganisms 4:36. https://doi.org/10.3390/microorganisms4030036
Kwiecinski J, Na M, Jarneborn A et al (2016) Tissue plasminogen activator coating on implant surfaces reduces Staphylococcus aureus biofilm formation. Appl Environ Microbiol 82:394–401. https://doi.org/10.1128/AEM.02803-15
Limoli DH, Jones CJ, Wozniak DJ (2015) Bacterial extracellular polysaccharides in biofilm formation and function. Microbiol Spectr. https://doi.org/10.1128/microbiolspec.MB-0011-2014
Eke PI, Rotimi VO, Odugbemi TO, Dosunmu-Ogunbi OO (1984) Effects of saliva and alpha-amylase on antibiotic sensitivity of bacteria. Afr J Med Med Sci 13:15–20
Craigen B, Dashiff A, Kadouri DE (2011) The use of commercially available alpha-amylase compounds to inhibit and remove Staphylococcus aureus biofilms. Open Microbiol J 5:21–31. https://doi.org/10.2174/1874285801105010021
Bradford C (2011) The use of commercially available alpha-amylase compounds to inhibit and remove Staphylococcus aureus biofilms. Open Microbiol J 5:21–31. https://doi.org/10.2174/1874285801105010021
Acknowledgements
The author would like to thank Nur Öztürk for her ongoing assistance as well as Altınbaş University Scientific Research Fund for their financial support.
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This research project was supported by Altınbaş University Scientific Research Fund. Project Number: PB2018-GÜZ-ECZ-7.
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All authors contributed to the concept and design of the study. Material preparation, data collection and analysis were performed by [NA], [CV], [CA], [BAA] and [TF]. [NA] wrote the first draft of the manuscript, and all contributors provided feedback on prior drafts. The final manuscript has been reviewed and approved by all writers.
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Aksoy, N., Vatansever, C., Adalı, C. et al. The Inhibitory Effects of Amylase and Streptokinase on Minimum Inhibitory Concentration of Antibiotics Used to Treat Gram Negative Bacteria Biofilm Infection on Indwelling Devices. Indian J Microbiol 63, 533–540 (2023). https://doi.org/10.1007/s12088-023-01109-1
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DOI: https://doi.org/10.1007/s12088-023-01109-1