Antibiotic Susceptibility Patterns and Virulence Profiles of Classical and Hypervirulent Strains Isolated from Clinical Samples in Khyber Pakhtunkhwa, Pakistan

Overview

Journal Pathogens

Date 2025 Jan 25

PMID 39861040

Authors

Azra

Taj Ali Khan

Ihtisham Ul Haq

Woranich Hinthong

Susana Campino

Aisha Gohar

Noman Khan

Muhammad Kashif

Ihsan Ullah

Taane G Clark

Affiliations

Soon will be listed here.

Abstract

The emergence of hypervirulent and carbapenem-resistant hypermucoviscous strains presents a significant public health challenge due to their increased virulence and resistance to multiple antibiotics. This study evaluates the antibiotic susceptibility patterns and virulence profiles of classical and hypervirulent strains isolated from various clinical samples. A total of 500 clinical samples were collected from patients at the Mardan Medical Complex and Ayub Medical Complex in KPK between July 2022 and June 2024. Among these, 64 strains were isolated and subsequently subjected to antimicrobial susceptibility testing (AST) and phenotypic virulence detection. Among the 64 isolates, 21 (32.8%) exhibited hypermucoviscosity, a characteristic associated with increased pathogenicity. Hemagglutination was observed in 35 (54.1%) of the isolates, indicating the presence of surface adhesins that facilitate bacterial adherence to host tissues. A high prevalence of biofilm formation was noted, with 54 (84%) isolates capable of forming biofilms, which are known to protect bacteria from antibiotics and the host immune response. Most isolates (59/64, 92.1%) were resistant against ampicillin, highlighting its limited efficacy against these strains. Conversely, the lowest resistance was observed for tigecycline, with only 15% (10/64) of the isolates showing resistance, indicating its potential utility as a treatment option. The study also found that 38 (59.3%) of the isolates were extended-spectrum beta-lactamase (ESBL) producers, 42 (65.6%) were multidrug-resistant (MDR), 32 (50%) were extensively drug-resistant (XDR), and 13 (20.3%) were resistant to carbapenems. The genetic study revealed biofilm producer and enhancer genes (, , , , , , and ) mainly in the hypervirulent strains. These hypervirulent strains also show a high number of resistance genes. The findings of this study underscore the critical need for the active surveillance of antimicrobial resistance and virulence determinants in . The coexistence of high levels of antibiotic resistance and virulence factors in these isolates poses a severe threat to public health, as it can lead to difficult-to-treat infections and increased morbidity and mortality.

References

Basak S, Singh P, Rajurkar M . Multidrug Resistant and Extensively Drug Resistant Bacteria: A Study. J Pathog. 2016; 2016:4065603. PMC: 4749793. DOI: 10.1155/2016/4065603. View

Li Y, Ni M . Regulation of biofilm formation in . Front Microbiol. 2023; 14:1238482. PMC: 10513181. DOI: 10.3389/fmicb.2023.1238482. View

Di Martino P, Cafferini N, Joly B, Darfeuille-Michaud A . Klebsiella pneumoniae type 3 pili facilitate adherence and biofilm formation on abiotic surfaces. Res Microbiol. 2003; 154(1):9-16. DOI: 10.1016/s0923-2508(02)00004-9. View

Yang D, Zhang Z . Biofilm-forming Klebsiella pneumoniae strains have greater likelihood of producing extended-spectrum beta-lactamases. J Hosp Infect. 2008; 68(4):369-71. DOI: 10.1016/j.jhin.2008.02.001. View

Fang R, Liu H, Zhang X, Dong G, Li J, Tian X . Difference in biofilm formation between carbapenem-resistant and carbapenem-sensitive Klebsiella pneumoniae based on analysis of mrkH distribution. Microb Pathog. 2021; 152:104743. DOI: 10.1016/j.micpath.2021.104743. View

Stahlhut S, Struve C, Krogfelt K, Reisner A . Biofilm formation of Klebsiella pneumoniae on urethral catheters requires either type 1 or type 3 fimbriae. FEMS Immunol Med Microbiol. 2012; 65(2):350-9. PMC: 3410544. DOI: 10.1111/j.1574-695X.2012.00965.x. View

Rahman S, Hameed A, Roghani M, Ullah Z . Multidrug resistant neonatal sepsis in Peshawar, Pakistan. Arch Dis Child Fetal Neonatal Ed. 2002; 87(1):F52-4. PMC: 1721434. DOI: 10.1136/fn.87.1.f52. View

Seemann T . Prokka: rapid prokaryotic genome annotation. Bioinformatics. 2014; 30(14):2068-9. DOI: 10.1093/bioinformatics/btu153. View

Zhang Y, Wang Q, Yin Y, Chen H, Jin L, Gu B . Epidemiology of Carbapenem-Resistant Enterobacteriaceae Infections: Report from the China CRE Network. Antimicrob Agents Chemother. 2017; 62(2). PMC: 5786810. DOI: 10.1128/AAC.01882-17. View

10.

Chew K, Lin R, Teo J . in Singapore: Hypervirulent Infections and the Carbapenemase Threat. Front Cell Infect Microbiol. 2018; 7:515. PMC: 5732907. DOI: 10.3389/fcimb.2017.00515. View

11.

Bjarnsholt T . The role of bacterial biofilms in chronic infections. APMIS Suppl. 2013; (136):1-51. DOI: 10.1111/apm.12099. View

12.

Wu M, Lin T, Hsieh P, Yang H, Wang J . Isolation of genes involved in biofilm formation of a Klebsiella pneumoniae strain causing pyogenic liver abscess. PLoS One. 2011; 6(8):e23500. PMC: 3155550. DOI: 10.1371/journal.pone.0023500. View

13.

Fatima S, Liaqat F, Akbar A, Sahfee M, Samad A, Anwar M . Virulent and multidrug-resistant Klebsiella pneumoniae from clinical samples in Balochistan. Int Wound J. 2021; 18(4):510-518. PMC: 8273605. DOI: 10.1111/iwj.13550. View

14.

Shadkam S, Goli H, Mirzaei B, Gholami M, Ahanjan M . Correlation between antimicrobial resistance and biofilm formation capability among Klebsiella pneumoniae strains isolated from hospitalized patients in Iran. Ann Clin Microbiol Antimicrob. 2021; 20(1):13. PMC: 7885248. DOI: 10.1186/s12941-021-00418-x. View

15.

Xu J, Zhao Z, Ge Y, He F . Rapid Emergence of a Pandrug-Resistant ST11 Isolate in an Inpatient in a Teaching Hospital in China After Treatment with Multiple Broad-Spectrum Antibiotics. Infect Drug Resist. 2020; 13:799-804. PMC: 7071855. DOI: 10.2147/IDR.S243334. View

16.

Merikallio H, Paakko P, Salmenkivi K, Kinnula V, Harju T, Soini Y . Expression of snail, twist, and Zeb1 in malignant mesothelioma. APMIS. 2012; 121(1):1-10. DOI: 10.1111/j.1600-0463.2012.02931.x. View

16.

Asri N, Ahmad S, Mohamud R, Mohd Hanafi N, Mohd Zaidi N, Irekeola A . Global Prevalence of Nosocomial Multidrug-Resistant : A Systematic Review and Meta-Analysis. Antibiotics (Basel). 2021; 10(12). PMC: 8698758. DOI: 10.3390/antibiotics10121508. View

17.

Vuotto C, Longo F, Pascolini C, Donelli G, Balice M, Libori M . Biofilm formation and antibiotic resistance in Klebsiella pneumoniae urinary strains. J Appl Microbiol. 2017; 123(4):1003-1018. DOI: 10.1111/jam.13533. View

18.

Page A, Cummins C, Hunt M, Wong V, Reuter S, Holden M . Roary: rapid large-scale prokaryote pan genome analysis. Bioinformatics. 2015; 31(22):3691-3. PMC: 4817141. DOI: 10.1093/bioinformatics/btv421. View

19.

Yu V, Hansen D, Ko W, Sagnimeni A, Klugman K, von Gottberg A . Virulence characteristics of Klebsiella and clinical manifestations of K. pneumoniae bloodstream infections. Emerg Infect Dis. 2008; 13(7):986-93. PMC: 2878244. DOI: 10.3201/eid1307.070187. View