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Current Pharmaceutical Design

Editor-in-Chief

ISSN (Print): 1381-6128
ISSN (Online): 1873-4286

Research Article

Lipase and Protease Production Ability of Multi-drug Resistant Bacteria Worsens the Outcomes of Wound Infections

Author(s): Attaur Rahman, Saiqa Sardar, Zeeshan Niaz, Asif Khan, Sheheryar Sheheryar*, Abdulwahed Fahad Alrefaei, Muhammad Hamayun and Sajid Ali*

Volume 30, Issue 17, 2024

Published on: 15 April, 2024

Page: [1307 - 1316] Pages: 10

DOI: 10.2174/0113816128302189240402043330

Abstract

Background: Surgical site infections are one of the major clinical problems in surgical departments that cost hundreds of millions of dollars to healthcare systems around the world.

Aim: The study aimed to address the pressing issue of surgical site infections, which pose significant clinical and financial burdens on healthcare systems globally. Recognizing the substantial costs incurred due to these infections, the research has focused on understanding the role of lipase and protease production by multi-drug resistant bacteria isolated from surgical wounds in the development of post-surgical wound infections.

Methods: For these purposes, 153 pus specimens were collected from patients with severe post-surgical wound infections having prolonged hospital stays. The specimens were inoculated on appropriate culture media. Gram staining and biochemical tests were used for the identification of bacterial growth on suitable culture media after 24 hours of incubation. The isolated pathogens were then applied for lipase and protease, key enzymes that could contribute to wound development, on tributyrin and skimmed milk agar, respectively. Following the CSLI guidelines, the Kirby-Bauer disc diffusion method was used to assess antibiotic susceptibility patterns. The results revealed that a significant proportion of the samples (127 out of 153) showed bacterial growth of Gram-negative (n = 66) and Gram-positive (n = 61) bacteria. In total, isolated 37 subjects were declared MDR due to their resistance to three or more than three antimicrobial agents. The most prevalent bacteria were Staphylococcus aureus (29.13%), followed by S. epidermidis (18.89%), Klebsiella pneumoniae (18.89%), Escherichia coli (14.96%), Pseudomonas aeruginosa (10.23%), and Proteus mirabilis (7.87%). Moreover, a considerable number of these bacteria exhibited lipase and protease activity with 70 bacterial strains as lipase positive on tributyrin agar, whereas 74 bacteria showed protease activity on skimmed milk agar with P. aeruginosa as the highest lipase (69.23%) and protease (76.92%) producer, followed by S. aureus (lipase 62.16% and protease 70.27%).

Results: The antimicrobial resistance was evaluated among enzyme producers and non-producers and it was found that the lipase and protease-producing bacteria revealed higher resistance to selected antibiotics than non-producers. Notably, fosfomycin and carbapenem were identified as effective antibiotics against the isolated bacterial strains. However, gram-positive bacteria displayed high resistance to lincomycin and clindamycin, while gram-negative bacteria were more resistant to cefuroxime and gentamicin.

Conclusion: In conclusion, the findings suggest that lipases and proteases produced by bacteria could contribute to drug resistance and act as virulence factors in the development of surgical site infections. Understanding the role of these enzymes may inform strategies for preventing and managing post-surgical wound infections more effectively.

Keywords: Lipase, protease, surgical site infections, antibiotic, drug resistance, bacteria.

[1]
Simone DB, Sartelli M, Coccolini F, et al. Intraoperative surgical site infection control and prevention: A position paper and future addendum to WSES intra-abdominal infections guidelines. World J Emerg Surg 2020; 15(1): 10.
[http://dx.doi.org/10.1186/s13017-020-0288-4] [PMID: 32041636]
[2]
Dessie W. Pattern of bacterial pathogens and their susceptibility isolated from surgical site infections at selected referral hospitals, Addis Ababa, Ethiopia. Int J Microbiol 2016; 2016: 2418902.
[http://dx.doi.org/10.1155/2016/2418902]
[3]
Abdul F. Staphylococcus epidermidis biofilms: Functional molecules; relation to virulence and the host immune response. J Life Sci Bio Sci Res 2021; 2(02): 42-53.
[http://dx.doi.org/10.38094/jlbsr20243]
[4]
Shettigar K, Murali TS. Virulence factors and clonal diversity of Staphylococcus aureus in colonization and wound infection with emphasis on diabetic foot infection. Eur J Clin Microbiol Infect Dis 2020; 39(12): 2235-46.
[http://dx.doi.org/10.1007/s10096-020-03984-8] [PMID: 32683595]
[5]
Lindsay S, Oates A, Bourdillon K. The detrimental impact of extracellular bacterial proteases on wound healing. Int Wound J 2017; 14(6): 1237-47.
[http://dx.doi.org/10.1111/iwj.12790] [PMID: 28745010]
[6]
Kon Y, Tsukada H, Hasegawa T, et al. The role of Pseudomonas aeruginosa elastase as a potent inflammatory factor in a rat air pouch inflammation model. FEMS Immunol Med Microbiol 1999; 25(3): 313-21.
[http://dx.doi.org/10.1111/j.1574-695X.1999.tb01356.x] [PMID: 10459586]
[7]
Saising J, Singdam S, Ongsakul M, Voravuthikunchai SP. Lipase, protease, and biofilm as the major virulence factors in staphylococci isolated from acne lesions. Biosci Trends 2012; 6(4): 160-4.
[http://dx.doi.org/10.5582/bst.2012.v6.4.160] [PMID: 23006962]
[8]
Chen X, Teoh WP, Stock MR, Resko ZJ, Alonzo F III. Branched chain fatty acid synthesis drives tissue-specific innate immune response and infection dynamics of Staphylococcus aureus. PLoS Pathog 2021; 17(9): e1009930.
[http://dx.doi.org/10.1371/journal.ppat.1009930] [PMID: 34496007]
[9]
Nassar MSM, Hazzah WA, Bakr WMK. Evaluation of antibiotic susceptibility test results: How guilty a laboratory could be? J Egypt Public Health Assoc 2019; 94(1): 4.
[http://dx.doi.org/10.1186/s42506-018-0006-1] [PMID: 30686832]
[10]
Bauer A. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 1966; 45(4): 493-6.
[http://dx.doi.org/10.1093/ajcp/45.4_ts.493]
[11]
Falagas ME, Koletsi PK, Bliziotis IA. The diversity of definitions of multidrug-resistant (MDR) and pandrug-resistant (PDR) Acinetobacter baumannii and Pseudomonas aeruginosa. J Med Microbiol 2006; 55(12): 1619-29.
[http://dx.doi.org/10.1099/jmm.0.46747-0] [PMID: 17108263]
[12]
Magiorakos AP, Srinivasan A, Carey RB, 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.
[http://dx.doi.org/10.1111/j.1469-0691.2011.03570.x] [PMID: 21793988]
[13]
Ilesanmi OI. Isolation, optimization and molecular characterization of lipase producing bacteria from contaminated soil. Sci Am 2020; 8: e00279.
[14]
Masi C, Gemechu G, Tafesse M. Isolation, screening, characterization, and identification of alkaline protease-producing bacteria from leather industry effluent. Ann Microbiol 2021; 71(1): 24.
[http://dx.doi.org/10.1186/s13213-021-01631-x]
[15]
Saravanakumar R, Devi PMB. Surgical site infection in a tertiary care centre-an overview - A cross sectional study. Int J Surg Open 2019; 21: 12-6.
[http://dx.doi.org/10.1016/j.ijso.2019.09.008]
[16]
Bastola R. Surgical site infections: Distribution studies of sample, outcome and antimicrobial susceptibility testing. J Med Microb Diagn 2017; 6(252): 2161-0703.1000252.
[http://dx.doi.org/10.4172/2161-0703.1000252]
[17]
Dessalegn L. Aerobic bacterial isolates from post-surgical wound and their antimicrobial susceptibility pattern: A hospital based cross-sectional study. J Med Res 2014; 3(2): 18-23.
[18]
Control CD. National nosocomial infections surveillance (NNIS) report, data summary from october 1986-April 1996, issued May 1996. Am J Infect Control 1996; 24(5): 380-8.
[http://dx.doi.org/10.1016/S0196-6553(96)90026-7] [PMID: 8902113]
[19]
Sterling T, Irwin JJ. ZINC 15-ligand discovery for everyone. J Chem Inf Model 2015; 55(11): 2324-37.
[http://dx.doi.org/10.1021/acs.jcim.5b00559] [PMID: 26479676]
[20]
Kumar P, Dalal V, Kotra DG, Kumar PI. In-silico approach to identify novel potent inhibitors against GraR of S. aureus. Front Biosci 2020; 25(7): 1337-60.
[http://dx.doi.org/10.2741/4859] [PMID: 32114436]
[21]
Morris GM, Huey R, Lindstrom W, et al. AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility. J Comput Chem 2009; 30(16): 2785-91.
[http://dx.doi.org/10.1002/jcc.21256] [PMID: 19399780]
[22]
Xue LC, Rodrigues JP, Kastritis PL, Bonvin AM, Vangone A. PRODIGY: A web server for predicting the binding affinity of protein-protein complexes. Bioinformatics 2016; 32(23): 3676-8.
[http://dx.doi.org/10.1093/bioinformatics/btw514] [PMID: 27503228]
[23]
Kurkcuoglu Z, Koukos PI, Citro N, et al. Performance of HADDOCK and a simple contact-based protein-ligand binding affinity predictor in the D3R Grand Challenge 2. J Comput Aided Mol Des 2018; 32(1): 175-85.
[http://dx.doi.org/10.1007/s10822-017-0049-y] [PMID: 28831657]
[24]
Dalal V, Dhankhar P, Singh V, et al. Structure-based identification of potential drugs against FmtA of Staphylococcus aureus: Virtual screening, molecular dynamics, MM-GBSA, and QM/MM. Protein J 2021; 40(2): 148-65.
[http://dx.doi.org/10.1007/s10930-020-09953-6] [PMID: 33421024]
[25]
Dalal V, Kumar P, Rakhaminov G, et al. Repurposing an ancient protein core structure: Structural studies on FmtA, a novel esterase of Staphylococcus aureus. J Mol Biol 2019; 431(17): 3107-23.
[http://dx.doi.org/10.1016/j.jmb.2019.06.019] [PMID: 31260692]
[26]
Singh V, Dhankhar P, Dalal V, Tomar S, Kumar P. In-silico functional and structural annotation of hypothetical protein from Klebsiella pneumonia: A potential drug target. J Mol Graph Model 2022; 116: 108262.
[http://dx.doi.org/10.1016/j.jmgm.2022.108262] [PMID: 35839717]
[27]
Babinski KJ, Ribeiro AA, Raetz CRH. The Escherichia coli gene encoding the UDP-2,3-diacylglucosamine pyrophosphatase of lipid A biosynthesis. J Biol Chem 2002; 277(29): 25937-46.
[http://dx.doi.org/10.1074/jbc.M204067200] [PMID: 12000770]
[28]
Lee M, Zhao J, Kwak SH, et al. Structure-activity relationship of sulfonyl piperazine LpxH inhibitors analyzed by an LpxE-coupled malachite green assay. ACS Infect Dis 2019; 5(4): 641-51.
[http://dx.doi.org/10.1021/acsinfecdis.8b00364] [PMID: 30721024]
[29]
Damale MG, Patil R, Ansari SA, et al. In-silico structure based drug design approach to find potential hits in ventilator-associated pneumonia caused by Pseudomonas aeruginosa. Comput Biol Med 2022; 146: 105597.
[http://dx.doi.org/10.1016/j.compbiomed.2022.105597] [PMID: 35751198]
[30]
Kalinin DV, Holl R. LpxC inhibitors: A patent review (2010-2016). Expert Opin Ther Pat 2017; 27(11): 1227-50.
[http://dx.doi.org/10.1080/13543776.2017.1360282] [PMID: 28742403]
[31]
Williams AH, Immormino RM, Gewirth DT, Raetz CRH. Structure of UDP-N-acetylglucosamine acyltransferase with a bound antibacterial pentadecapeptide. Proc Natl Acad Sci 2006; 103(29): 10877-82.
[http://dx.doi.org/10.1073/pnas.0604465103] [PMID: 16835299]
[32]
Jenkins RJ, Dotson GD. Dual targeting antibacterial peptide inhibitor of early lipid A biosynthesis. ACS Chem Biol 2012; 7(7): 1170-7.
[http://dx.doi.org/10.1021/cb300094a] [PMID: 22530734]
[33]
Dangkulwanich M, Raetz CRH, Williams AH. Structure guided design of an antibacterial peptide that targets UDP-N-acetylglucosamine acyltransferase. Sci Rep 2019; 9(1): 3947.
[http://dx.doi.org/10.1038/s41598-019-40418-8] [PMID: 30850651]
[34]
Arif SM, Floto RA, Blundell TL. Using  structure-guided  fragment-based drug discovery to target Pseudomonas aeruginosa infections in cystic fibrosis. Front Mol Biosci 2022; 9: 857000.
[http://dx.doi.org/10.3389/fmolb.2022.857000] [PMID: 35433835]
[35]
Silver LL. Multi-targeting by monotherapeutic antibacterials. Nat Rev Drug Discov 2007; 6(1): 41-55.
[http://dx.doi.org/10.1038/nrd2202] [PMID: 17159922]
[36]
Qin Z, Zhang J, Xu B, et al. Structure-based discovery of inhibitors of the YycG histidine kinase: New chemical leads to combat Staphylococcus epidermidis infections. BMC Microbiol 2006; 6(1): 96.
[http://dx.doi.org/10.1186/1471-2180-6-96] [PMID: 17094812]
[37]
McCann MT, Gilmore BF, Gorman SP. Staphylococcus epidermidis device-related infections: Pathogenesis and clinical management. J Pharm Pharmacol 2008; 60(12): 1551-71.
[http://dx.doi.org/10.1211/jpp/60.12.0001] [PMID: 19000360]
[38]
Paharik AE, Horswill AR. The staphylococcal biofilm: Adhesins, regulation, and host response. Virulence mechanisms of bacterial pathogens. Wiley 2016; pp. 529-66.
[http://dx.doi.org/10.1128/9781555819286.ch19]
[39]
Alonso GIS, García MS, Cabrera BG, et al. Low concentration of the neutrophil proteases Cathepsin G, Cathepsin B, proteinase-3 and metalloproteinase-9 induce biofilm formation in non-biofilm-forming Staphylococcus epidermidis Isolates. Int J Mol Sci 2022; 23(9): 4992.
[http://dx.doi.org/10.3390/ijms23094992] [PMID: 35563384]
[40]
Dubin G. Extracellular proteases of Staphylococcus spp. Biol Chem 2002; 383(7-8): 1075-2086.
[http://dx.doi.org/10.1515/BC.2002.116]
[41]
Sethi G, Hwang JH, Krishna R. Structure based exploration of potential lead molecules against the extracellular cysteine protease (EcpA) of Staphylococcus epidermidis: A therapeutic halt. J Biomol Struct Dyn 2023; 2023: 1-17.
[http://dx.doi.org/10.1080/07391102.2023.2250455] [PMID: 37615425]
[42]
Zou L, Evans CR, Do VD, Losefsky QP, Ngo DQ, McGillivray SM. Loss of the ClpXP protease leads to decreased resistance to cell-envelope targeting antimicrobials in Bacillus anthracis sterne. Front Microbiol 2021; 12: 719548.
[http://dx.doi.org/10.3389/fmicb.2021.719548] [PMID: 34497598]
[43]
Devanathan K. Correlation of drug resistance pattern with lipase production in clinical isolates of Klebsiella pneumoniae. Int J Eng Res 2022; 11(5): 18-21.
[44]
Hinz A, Lee S, Jacoby K, Manoil C. Membrane proteases and aminoglycoside antibiotic resistance. J Bacteriol 2011; 193(18): 4790-7.
[http://dx.doi.org/10.1128/JB.05133-11] [PMID: 21764915]
[45]
Sedarat Z, Robinson TAW. Biofilm formation by pathogenic bacteria: Applying a Staphylococcus aureus model to appraise potential targets for therapeutic intervention. Pathogens 2022; 11(4): 388.
[http://dx.doi.org/10.3390/pathogens11040388] [PMID: 35456063]
[46]
Proma TT, Ahmed T. Evaluation of drug resistance before and after biofilm formation of bacteria causing wound infection and detection of their protease activity. Int J Infect 2021; 8(3): e108247.
[http://dx.doi.org/10.5812/iji.108247]
[47]
Prasad ASB, Shruptha P, Prabhu V, et al. Pseudomonas aeruginosa virulence proteins pseudolysin and protease IV impede cutaneous wound healing. Lab Invest 2020; 100(12): 1532-50.
[http://dx.doi.org/10.1038/s41374-020-00478-1] [PMID: 32801335]

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