Biofilm Production and Porin Permeability Activity in Clinical Isolates of Ciprofloxacin-Resistant Klebsiella pneumoniae in the Tertiary Hospital in Purwokerto, Indonesia
DOI:
https://doi.org/10.48048/tis.2024.7580Keywords:
Antibiotic resistance, Biofilm, Ciprofloxacin, Klebsiella pneumoniae, PorinAbstract
World Health Organization reports that the current use of ciprofloxacin as a broad-spectrum antibiotic shows a pattern of high bacterial resistance in many countries. Klebsiella pneumoniae is known as a Gram-negative bacterium that most often causes infection and is resistant to antibiotics. This study aimed to find patterns of resistance mechanisms based on biofilm production and porin permeability activity in K. pneumoniae clinical specimens from patients at RSUD Prof. Dr Margono Soekardjo Purwokerto, Indonesia. Several isolates of ciprofloxacin-resistant K. pneumoniae were isolated from clinical specimens of blood, sputum, urine, pus, stool and pleural fluid from August to October 2022. Identification and sensitivity testing of K. pneumoniae to ciprofloxacin were performed using Vitek® 2 Compact. A test for biofilm production is carried out by measuring optical density (OD) with a microplate at a wavelength of 630 nm. The porin permeability activity was determined using the value of the minimum inhibitory concentration (MIC) by the LC-MS spectrophotometry method at a wavelength of 630 nm. The results showed that 72 (47 %) isolates were resistant to ciprofloxacin. Among them, 41.3 % (24/58) of ciprofloxacin-resistant K. pneumoniae were able to produce strong biofilm. Porin permeability is indicated by MIC values of 1, 2 and 4 mg/L with bacterial cell counts of 32×106, 19×106 and 34×106 CFU/mL, respectively. This number decreased from the initial control with a bacterial cell count of 10.15×107 CFU/mL. Analysis of the correlation between the significance of biofilm production and the number of bacterial cells indicated that biofilm formation was related to the number of bacterial cells (R = 0.628, p ≤ 0.05). In conclusion, the resistance mechanism of ciprofloxacin-resistant K. pneumoniae involves biofilm production and decreased porin permeability activity.
HIGHLIGHTS
- Major resistance mechanisms in pneumoniae decreased cell permeability from porin activity and biofilm formation
- Bacterial outer membrane permeability to antimicrobial substances influences inherent resistance. Bacteria produce biofilm to increase their resistance to antibiotics. Porins are non-specific diffusion transport proteins that play a part in the antibiotic resistance process
- The evolutionary development in the case of antibiotic resistance has an impact on the emergence of understanding in new perspective patterns on how to overcome health problems related to antibiotic resistance
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RF Alvi, B Aslam, MH Rasool, S Muzammil, AB Siddique, N Yasmeen, M Khurshid, N Sarwar, A Almatroudi, R Hussain and Z Baloch. Transcriptional response of multidrug-resistant Klebsiella pneumoniae clinical isolates to ciprofloxacin stress. Can. J. Infec. Dis. Med. Microbiol. 2021; 2021, 5570963.
SS Malekshahi, J Yavarian, NZ Shafiei-Jandaghi, T Mokhtari-Azad and M Farahmand. Prevalence of human metapneumovirus infections in Iran: A systematic review and meta-analysis. Fetal Pediatr. Pathol. 2021; 40, 663-73.
D Moradigaravand, V Martin, SJ Peacock and J Parkhill. Evolution and epidemiology of multidrug-resistant Klebsiella pneumoniae in the United Kingdom and Ireland. mBio 2017; 8, e01976-16.
T Banerjee, A Mishra, A Das, S Sharma, H Barman and G Yadav. High prevalence and endemicity of multidrug resistant Acinetobacter spp. in intensive care unit of a tertiary care hospital, Varanasi, India. J. Pathogens 2018; 2018, 9129083.
B Allegranzi, E Tartari and D Pittet. Seconds save lives e clean your hands: The 5 May 2021 World Health Organization SAVE LIVES: Clean Your Hands campaign. J. Hosp. Infect. 2021; 111, 1-3.
A Custovic, J Smajlovic, S Hadzic, S Ahmetagic, N Tihic and H Hadzagic. Epidemiological surveillance of bacterial nosocomial infections in the surgical intensive care unit. Mater. Soc. Med. 2014; 26, 7-11.
SP Singh, N Yaduvanshi, C Sahu, S Singh, A Agarwal and U Ghoshal. Epidemiology, antimicrobial susceptibility patterns and outcomes of bacteremia in an Apex trauma center of a tertiary health care institute with special reference to Methicillin Resistant Staphylococcus aureus (MRSA): A Prospective Cohort study. Int. J. Med. Sci. Curr. Res. 2021; 4, 435-43.
TC Hendrik, AFIH Voor and MC Vos. Clinical and molecular epidemiology of producing klebsiella spp.: A systematic review and meta-analyses. PLos One 2015; 10, e0140754.
M Bassetti, E Righi, A Carnelutti, E Graziano and A Russo. Multidrug-resistant klebsiella pneumoniae: Challenges for treatment, prevention and infection control. Expet. Rev. Anti Infective Ther. 2018; 16, 749-61.
O Unlu, BR Ersoz, AI Tosun and M Demirci. Epidemic Klebsiella pneumoniae ST258 incidence in ICU patients admitted to a university hospital in Istanbul. J. Infect. Develop. Countries 2021; 15, 665-71.
AA Al-Naqshbandi, MA Chawsheen and HH Abdulqader. Prevalence and antimicrobial susceptibility of bacterial pathogens isolated from urine specimens received in rizgary hospital - Erbil. J. Infect. Publ. Health 2019; 12, 330-6.
P Kazanjian. History of antimicrobial stewardship. In: K LaPlante, C Cunha, H Morrill, L Rice and E Mylonakis (Eds.). Antimicrobial stewardship: Principles and practice. CABI, Oxfordshire, 2017.
KL Wyres and KE Holt. Klebsiella pneumoniae as a key trafficker of drug resistance genes from environmental to clinically important bacteria. Curr. Opin. Microbiol. 2018; 45, 131-9.
K Starzyk-Łuszcz, TM Zielonka, J Jakubik and K Życińska. Mortality due to nosocomial infection with Klebsiella pneumoniae ESBL+. Adv. Exp. Med. Biol. 2017; 1022, 19-26.
LX Su, XT Wang, P Pan, WZ Chai and DW Liu. Infection management strategy based on prevention and control of nosocomial infections in intensive care units. Chin. Med. J. 2019; 132, 115-9.
L Burke, H Humphreys and D Fitzgerald-Hughes. The revolving door between hospital and community: Extended-spectrum beta-lactamase-producing Escherichia coli in Dublin. J. Hosp. Infect. 2012; 81, 192-8.
P Nahar, L Unicomb, PJ Lucas, MR Uddin, MA Islam, FA Nizame, N Khisa, SMS Akter and EK Rousham. What contributes to inappropriate antibiotic dispensing among qualified and unqualified healthcare providers in Bangladesh? A qualitative study. BMC Health Serv. Res. 2020; 20, 656.
B Mehrad, NM Clark, GG Zhanel and JP Lynch. Antimicrobial resistance in hospital-acquired gram-negative bacterial infections. Chest 2015; 147, 1413-21.
O Zlatian, AT Balasoiu, M Balasoiu, O Cristea, AO Docea, R Mitrut, DA Spandidos, AM Tsatsakis, G Bancescu and D Calina. Antimicrobial resistance in bacterial pathogens among hospitalised patients with severe invasive infections. Exp. Ther. Med. 2018; 16, 4499-510.
D Baggio and MR Ananda-Rajah. Fluoroquinolone antibiotics and adverse events. Aust. Prescriber 2021; 44, 161-4.
R Handal, L Qunibi, I Sahouri, M Juhari, R Dawodi, H Marzouqa and M Hindiyeh. Characterization of carbapenem-resistant Acinetobacter baumannii strains isolated from hospitalized patients in Palestine. Int. J. Microbiol. 2006; 2017, 8012104.
World Health Organization. The WHO list of Critically Important Antimicrobials (CIA). World Health Organization, Switzerland, 2019.
World Health Organization (WHO), GLASS Report: Early Implementation 2020, 2020. https://www.who.int/publications/i/item/9789240005587
FSA Al-Mayahi. A preliminary study of Aminoglycoside Modifying Enzymes (AMEs) of Multiple Antibiotic Resistance of Methicillin-resistant Staphylococcus aureus (MRSA) isolated from clinical specimens in Al-Diwaniya/Iraq. Jordan J. Biol. Sci. 2021; 14, 733-41.
JD Prajapati, U Kleinekath and M Winterhalter. How to enter a bacterium: Bacterial porins and the permeation of antibiotics. Chem. Rev. 2021; 121, 5158-92.
H Nirwati, K Sinanjung, F Fahrunissa, F Wijaya, S Napitupulu, VP Hati, MS Hakim, A Meliala, AT Aman and T Nuryastuti. Biofilm formation and antibiotic resistance of Klebsiella pneumoniae isolated from clinical samples in a tertiary care hospital, Klaten, Indonesia. BMC Proc. 2019; 13, 20.
J Arachchige, A Sampath and M Kothalawala. Use of high-dose ciprofloxacin for recurrent biofilm-forming multidrug-resistant Klebsiella pneumoniae bacteremia. Germs 2021; 11, 449-53.
E Paluch, J Rewak-Soroczyńska, I Jędrusik, E Mazurkiewicz and K Jermakow. Prevention of biofilm formation by quorum quenching. Appl. Microbiol. Biotechnol. 2020; 104, 1871-81.
T Mah. Biofilm-specific antibiotic resistance. Future Microbiol. 2012; 7, 1061-72.
L Surgers, A Boyd, PM Girard, G Arlet and D Decré. Biofilm formation by ESBL-producing strains of Escherichia coli and Klebsiella pneumoniae. Int. J. Med. Microbiol. 2019; 309, 13-8.
V Cepas, Y López, E Muñoz, D Rolo, C Ardanuy, S Martí, M Xercavins, JP Horcajada, J Bosch and SM Soto. Relationship between biofilm formation and antimicrobial resistance in Gram-negative bacteria. Microb. Drug Resist. 2019; 25, 72-9.
E Christaki, M Marcou and A Tofarides. Antimicrobial resistance in bacteria: Mechanisms, evolution, and persistence. J. Mol. Evol. 2019; 88, 26-40.
SS Walker and TA Black. Are outer-membrane targets the solution for MDR Gram-negative bacteria? Drug Discov. Today 2021; 26, 2152-8.
Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing: 25th Informational supplement. Clinical and Laboratory Standards Institute, Pennsylvania, 2015.
A Hassan, J Usman, F Kaleem, M Omair, A Khalid and M Iqbal. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz. J. Infect. Dis. 2011; 15, 305-11.
B Kowalska-Krochmal and R Dudek-Wicher. The minimum inhibitory concentration of antibiotics: Methods, interpretation, clinical relevance. Pathogens 2021; 10, 165.
X Qin, S Wu, M Hao, J Zhu, B Ding, Y Yang, X Xu, M Wang, F Yang and F Hu. The colonization of carbapenem-resistant klebsiella pneumoniae: Epidemiology, resistance mechanisms, and risk factors in patients admitted to intensive care units in China. J. Infect. Dis. 2020; 221, S206-S214.
HZ Hamdan, E Kubbara, AM Adam, OS Hassan, SO Suliman and I Adam. Urinary tract infections and antimicrobial sensitivity among diabetic patients at Khartoum, Sudan. Ann. Clin. Microbiol. Antimicrobials 2015; 14, 26.
A Kaur, RK Wasan, C Kaur, P Sethi and V Kaur. Antibiotic resistance pattern of Klebsiella pneumoniae a major problem for society. Int. J. Health Sci. 2022; 6, 4699-712.
R Osagie, A Eyaufe, O Iserhienrhien, M Okodua, F Unuabonah and O Daibo. Antibiotic susceptibility profile of Klebsiella pneumoniae isolated from sputum samples amongst hospitalized adults in parts of Edo State, South-South, Nigeria. Merit Res. J. Med. Med. Sci. 2017; 5, 378-83.
J Akter, AMMA Chowdhury and MA Forkan. Study on prevalence and antibiotic resistance pattern of Klebsiella isolated from clinical samples in south east region of Bangladesh. Am. J. Drug Discov. Dev. 2014; 4, 73-9.
K Shilpa, R Thomas and A Ramyshree. Isolation and Antimicrobial sensitivity pattern of Klebsiella pneumoniae from sputum samples in a tertiary care hospital. Int. J. Biomed. Adv. Res. 2016; 7, 53-7.
M Melzer, I Petersen and T Cheasty. The difference in serotypes between extended-β-lactamase (ESBL) and non-ESBL-producing E. coli blood culture isolates at a UK district general hospital. J. Hosp. Infect. 2008; 68, 367-9.
JX Zheng, ZW Lin, C Chen, Z Chen, FJ Lin, Y Wu, SY Yang, X Sun, WM Yao, DY Li, ZJ Yu, JL Jin, D Qu and QW Deng. Biofilm formation in Klebsiella pneumoniae bacteremia strains was found to be associated with CC23 and the presence of wcaG. Front. Cell. Infect. Microbiol. 2018; 8, 21.
O Favour, F Osazuwa, RM Mordi, E Osazuwa, SS Taiwo, OAT Alli, DO Ogbolu, EO Akanni and KC Anukam. Klebsiella has taken lead among uropathogens in University of Benin Teaching Hospital, Benin City, Nigeria-An observation. New York Sci. J. 2014; 3, 61-4.
C Wang, Z Yuan, W Huang, L Yan, J Tang and CW Liu. Epidemiologic analysis and control strategy of Klebsiella pneumoniae infection in intensive care units in a teaching hospital of People’s Republic of China. Infect. Drug Resist. 2019; 12, 391-8.
M Ahmad, AB Siddique, S Muzammil, M Shafique, Z Nawaz, M Khurshid, MH Rasool, MM Jalees, N Sarwar and B Aslam. Occurrence of hypervirulent Klebsiella pneumoniae in clinical settings and lytic potential of bacteriophages against the isolates. Jundishapur J. Microbiol. 2022; 15, e120027.
AS Moini, B Soltani, AT Ardakani, A Moravveji, M Erami, MH Rezaei and M Namazi. Multidrug-resistant Escherichia coli and Klebsiella pneumoniae isolated from patients in Kashan, Iran. Jundishapur J. Microbiol. 2015; 8, e27517.
M Walaszek, A Rozanka, MZ Wałaszek and J Wójkowska-Mach. Epidemiology of Ventilator-Associated Pneumonia, microbiological diagnostics and the length of antimicrobial treatment in the Polish Intensive Care Units in the years 2013 - 2015. BMC Infect. Dis. 2018; 18, 308.
GM Adwan, DM Owda and AA Abu-Hijleh. Prevalence of capsular polysaccharide genes and antibiotic resistance pattern of Klebsiella pneumoniae in Palestine. Jordan J. Biol. Sci. 2020; 13, 475-82.
JH Lee. Perspectives towards antibiotic resistance: From molecules to population. J. Microbiol. 2019; 57, 181-4.
F Hu, Y Yang, Y Zheng, S Wu, X Jiang, D Zhu and F Wang. Resistance reported from China antimicrobial surveillance network (CHINET) in 2018. Eur. J. Clin. Microbiol. Infect. Dis. 2019; 38, 2275-81.
KJ Aldred, RJ Kerns and N Oshero. Mechanism of quinolone action and resistance. Biochemistry 2014; 53, 1565-74.
XY Zhou, XG Ye, LT He, SR Zhang, RL Wang, J Zhou and ZS He. In vitro characterization and inhibition of the interaction between ciprofloxacin and berberine against multidrug-resistant Klebsiella pneumoniae. J. Antibiot. 2016; 69, 741-6.
S Saha, KM Devi, S Damrolien, KS Devi, Krossnunpuii and KT Sharma. Biofilm production and its correlation with antibiotic resistance pattern among clinical isolates of Pseudomonas aeruginosa in a tertiary care hospital in north-east India. Int. J. Adv. Med. 2018; 5, 964-8.
MT Al-Ouqaili, SQ Al-Quhli and MY Al-Izzy. The role of milleri Streptococci in the formation of cariogenic biofilm: Bacteriological aspects. Jordan J. Biol. Sci. 2011; 4, 165-72.
W Chen, B Li, S Li, YW Ou and Q Ou. Effects of scutellaria baicalensis on activity and biofilm formation of Klebsiella pneumoniae. Chin. Med. Sci. J. 2016; 31, 180-4.
CN Murphy and S Clegg. Klebsiella pneumoniae and type 3 fimbriae: Nosocomial infection, regulation and biofilm formation. Future Microbiol. 2012; 7, 991-1002.
A Cherif-Antar, B Moussa-Boudjemâa, N Didouh, K Medjahdi, B Mayo and AB Flórez. Diversity and biofilm-forming capability of bacteria recovered from stainless steel pipes of a milk-processing dairy plant. Dairy Sci. Tech. 2016; 96, 27-38.
A Grillon, F Schramm, M Kleinberg and F Jehl. Comparative activity of ciprofloxacin, levofloxacin and moxifloxacin against Klebsiella pneumoniae, Pseudomonas aeruginosa and Stenotrophomonas maltophilia assessed by minimum inhibitory concentrations and time-kill studies. PLoS One 2016; 11, e0156690.
SK Tiwari, S Wang, Y Huang, X Zhou, HHK Xu, B Ren, X Peng, Y Xiao, M Li and L Cheng. Starvation survival and biofilm formation under subminimum inhibitory concentration of QAMs. Biomed. Res. Int. 2021; 2021, 8461245.
Y Pu, Y Ke and F Bai. Active efflux in dormant bacterial cells - New insights into antibiotic persistence. Drug Resist. Updates 2017; 30, 7-14.
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