Int J Pharm Pharm Sci, Vol 17, Issue 6, 12-14Original Article

BURDEN OF KLEBSIELLA PNEUMONIAE AND ESCHERICHIA COLI ISOLATES THAT PRODUCE EXTENDED-SPECTRUM BETA-LACTAMASES AT THE TERTIARY CARE HOSPITAL IN UTTAR PRADESH, LUCKNOW

NEETI MISHRA*, DAYAVANTI KUMARI, KHYATI TIWARI

Department of Microbiology, T. S. Misra Medical College and Hospital Uttar Pradesh, India
*Corresponding author: Neeti Mishra; *Email: neetitripathi26@yahoo.com

Received: 07 Apr 2025, Revised and Accepted: 08 May 2025

ABSTRACT

Objective: To ascertain the prevalence of Escherichia coli isolates and Klebsiella pneumoniae that generates Extended-Spectrum Beta-Lactamases (ESBLs) in the tertiary care hospital in Lucknow, Uttar Pradesh.

Methods: Screening tests have been developed by the Clinical Laboratory Standard Institute (CLSI) to identify the Klebsiellla pneumoniae and Escherichia coli that generate ESBLs. Finally, performing combination disk test on probable ESBL-producing isolates, ESBL phenotypic confirmation was established.

Results: 94 (62.7%) of the 150 ESBL-positive isolates were female, while 56 (37.3%) were male. The largest concentration of E. coli which was positive for ESBL production was from urine, and it also shows the lowest concentrations coming from sputum, blood, and CSF samples and the largest concentration of ESBL-positive Klebsiella pneumoniae was found in sputum, followed by urine, pus, and blood, while the lowest concentration was found in a CSF sample. E. coli had 158 of the 267 isolates, and of them, 80 were ESBL positive. Similarly, 70 of the 109 Klebsiella pneumoniae isolates tested were ESBL positive.

Conclusion: E. coli had an ESBL prevalence of 53.34% in the current study, while K. pneumoniae had 46.7% prevalence. Klebsiella pneumoniae and Escherichia coli that produce ESBLs were found in relatively high numbers in the urine, sputum, pus, ET aspirate, and blood samples. The doctors must follow stringent infection control procedures at the hospital and judicious antimicrobial usage guidelines.

Keywords: Extended-spectrum beta-lactamase, Beta-lactamase

© 2025 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/)
DOI: https://dx.doi.org/10.22159/ijpps.2025v17i6.54499 Journal homepage: https://innovareacademics.in/journals/index.php/ijpps

INTRODUCTION

The most typical method of treating bacterial infections is using β-lactam antimicrobials, which also continue to be the main source of β-lactam antibiotic resistance among Gram-negative bacteria globally. Numerous beta-lactams have been continually exposed to different bacterial strains, resulting in dynamic, on-going creation and mutation of enzymes in these bacteria. This has increased these bacteria's activity even against recently developed beta-lactam antibiotics. Extended-spectrum β-lactamases (ESBLs) are the name given to these enzymes [1-3]. The various drug resistances of these organisms make treatment difficult. Due to a number of factors, including the difficulties in detecting ESBL production and inconsistent reporting, it is challenging to quantify the occurrence of ESBL-producing organisms at the level of a larger geographic scale [4]. Recent surveys revealed a marked rise in the rate of ESBLs everywhere in the world [5-12]. The Clinical Laboratory Standard Institute (CLSI) recommends routine reporting and testing for the main infections that continue to be found worldwide that produce ESBLs: Klebsiella pneumoniae and Escherichia coli [13, 14]. ESBL prevalence varies from institution to institution. Previous studies from India and internationally suggest that there might be anywhere from 8 to 80% production of ESBL. However, the antimicrobial resistance patterns and the frequency of ESBL production in Klebsiella pneumoniae isolates have not been extensively studied. Therefore, the primary goal of the current investigation is to determine the prevalence of Klebsiella pneumoniae and Escherichia coli isolates that produce ESBLs in the tertiary care hospital.

MATERIALS AND METHODS

The Institutional Ethics Committee (IEC) gave its clearance for the study, which was carried out in the Microbiology Department at T. S. M. Medical College, Lucknow, U. P., India, from January 2018 to February 2020. (Reference No. TSMMC and H/STC/952/2021)

Inclusion criteria

Only those patients who gave their permission and agreed to take part in the study.

Exclusion criteria

Individuals who are not interested in participating in the study and those who have systemic illness.

Data collection

The prospective investigation was conducted by the Microbiology Department of T. S. Misra Medical College and Hospital located in Amausi, Lucknow, Uttar Pradesh. A total of 384 different clinical samples were gathered in sterile containers that came from various departments. Samples that included isolated Klebsiella pneumoniae and E. coli were used in this experiment. All samples-whether from outpatient or inpatient—were included during the study period. Each patient's Performa was filled out after a thorough history was gathered, including information on age, sex, and medical history. According to CLSI recommendations, ESBL detection was carried out.

Processing of clinical samples

All clinical samples were cultured on routine culture media and then processed after an overnight incubation period at 37 °C. For the blood sample, brain heart infusion broth was incubated at 37 °C overnight. A drop of the broth from the Brain Heart Infusion was employed as an inoculum on routine culture media. These agars were then incubated overnight at 37 °C. The blood sample was deemed negative if bacterial colonies were not visible after seven days. Only isolates of Klebsiella pneumoniae and Escherichia coli that were collected from clinical specimens were taken into account in this experiment as a pure and predominant growth. The biochemical reactions and colony morphology of the organisms were used to identify them [15].

Screening for ESBL production

Screening tests have been developed by the CLSI to identify the Klebsiella pneumoniae and E. coli that generate ESBLs [14]. If a strain showed an inhibitory zone of ≤22 mm for Ceftazidime, ≤27 mm for Cefotaxime, and ≤25 mm for Ceftriaxone, it was chosen for confirmatory ESBL testing based on CLSI criteria.

ESBL production confirmation test

Phenotypic confirmation by combination disk test [16]. By performing phenotypic tests on probable ESBL-producing isolates, ESBL production was established. A lawn culture of the inoculum was created, and 25 mm-apart discs of Ceftazidime (30 μg) and Ceftazidime+Clavulanic acid (30 μg+10 μg) were used. Ceftazidime+Clavulanic acid, as ESBL makers, demonstrated about a 5 mm increase in the zone of inhibition in comparison to Ceftazidime alone. Isolates of ESBLs that were Cefoxitin-resistant were not considered for the investigation. This is done in order to eliminate related Amp C beta-lactamases [17, 18].

RESULTS

From January 2019 to December 2021, 384 clinical samples in total were examined for culture and sensitivity. Out of these, 267 samples demonstrated growth of Klebsiella pneumoniae and E. coli. Table 1 shows the gender-wise distribution of ESBL producers and ESBL non-producers in clinical isolates. 94 (62.7%) of the 150 ESBL-positive isolates were female, while 56 (37.3%) were male.

Table 1: Gender wise distribution

Gender ESBL producer Non-ESBL producer Total
Female 94 72 166
Male 56 45 101
Total 150 117 267

Table 2: Dispersal pattern of the clinical isolates

Type of samples Total samples Escherichia coli Escherichia coli positive for ESBL Klebsiella pneumoniae Klebsiella pneumoniae positive for ESBL Total
Mid-stream urine 64 28 16 22 12 50
Sputum 79 38 14 19 14 57
Pus 58 16 09 16 10 32
CSF 15 02 01 06 05 08
Blood 47 23 12 11 09 34
Endotracheal aspirate 38 18 10 10 07 28
Body fluids 44 19 08 13 08 32
High vaginal swab 39 14 10 12 05 26

Table 2 indicates that the largest concentration of E. coli that was positive for ESBL production was from urine, and it also shows the lowest concentrations coming from sputum, blood, and CSF samples. The largest concentration of ESBL-positive Klebsiella pneumoniae was found in sputum, followed by urine, pus, and blood, while the lowest concentration was found in a CSF sample.

Table 3: ESBL prevalence

Organism Total ESBL
Klebsiella pneumoniae 109 80 (53.34%)
E. coli 158 70 (46.66%)
Total 267 150

Prevalence of ESBL is described in table 3. E. coli had 158 of the 267 isolates, and of them, 80 were ESBL positive. Similarly, 70 of the 109 Klebsiella pneumoniae isolates tested were ESBL positive.

DISCUSSION

Many scientific studies have examined the incidence of K. pneumoniae strains that produce ESBL, for example, in India, where many groups have documented a significant burden of these pathogenic strains. In the current investigation, the prevalence of ESBLs in E. coli was 53.34%, while that in Klebsiella pneumoniae was 46.66%. The percentage of bacteria that produce ESBLs in India varied from 4% to 83% [19, 20]. Maharashtra has a lower proportion of ESBL producers, according to Rodrigues et al. [21]. Out of 47 K. pneumoniae isolates, four (8.5%) were positive ESBL producers, according to their findings. At this moment, the production of ESBL should have increased in the same location, and this ratio most likely indicates their early stages. This makes sense because there are a number of variables that affect the frequency of extended-spectrum beta-lactamase producers in hospitals, such as the institution's antibiotic policies, the infection rates among staff, and the types of disinfectants used, particularly in the intensive care unit [22]. ESBL isolates were more prevalent in females in the current study. Gupta S. et al. provides similar results [23]. Studies by Mendelson G et al. and Bazzaz B et al. indicate that males are more susceptible to ESBL production [24, 25] ESBL-producing Klebsiella species have also been observed in recent years in the USA (42-44%) and Canada (4.9%), Turkey (78.6%), Spain (20.8%), Taiwan (28.4%), Algeria (20%), and China (51%) [26]. There are significant geographic variations in the occurrence of ESBLs when targeting the epidemiology in Europe. According to recent research involving 1,610 E. coli and 785 K. pneumoniae isolates from 31 sites across ten European countries, the incidence of ESBL in these organisms varied significantly, ranging from 1.5% in Germany to 39–47% in Russia, Poland, and Turkey [27]. E. coli had an ESBL prevalence of 53.34%, and K. pneumoniae had 46.7% prevalence. In comparison to an Indian investigation [28], this found that about 40% of urine isolates of E. coli and K. pneumoniae were ESBL positive, this value was noticeably higher. Latin America (54.4%), the western Pacific (24.6%), and Europe (22.6%) were the regions with the highest reported isolation rates of K. pneumoniae generating ESBLs. 8.5%, 7.8%, and 5.3%, respectively, of E. coli in these regions were observed to produce ESBLs.

LIMITATIONS OF THE STUDY

ESBL production's molecular characterisation could not be investigated because of the lack of resources. The participants in this study were restricted to one hospital. The results might therefore not be generalizable to other regions.

CONCLUSION

E. coli had an ESBL prevalence of 53.34% in the current study, while K. pneumoniae had 46.7% prevalence. Klebsiella pneumoniae and Escherichia coli that produce ESBLs were found in relatively high numbers in the urine, sputum, pus, ET aspirate, and blood samples. The doctors must follow stringent infection control procedures at the hospital and judicious antimicrobial usage guidelines. Clinical laboratories must regularly and routinely monitor the presence of clinical isolates that produce ESBLs.

FUNDING

Nil

AUTHORS CONTRIBUTIONS

Drs. Neeti and Daya were responsible for creating the conceptual framework, the draft, and the data analysis. As Dr. Khyati wrote the manuscript and oversaw the final round of editing, Dr. Daya also helped with data collection and analysis.

CONFLICT OF INTERESTS

Declared none

REFERENCES

  1. Livermore DM. Bacterial resistance: origins, epidemiology and impact. Clin Infect Dis. 2003 Jan 15;36 Suppl 1:S11-23. doi: 10.1086/344654, PMID 12516026.

  2. Paterson DL, Bonomo RA. Extended spectrum beta-lactamases: a clinical update. Clin Microbiol Rev. 2005 Oct;18(4):657-86. doi: 10.1128/CMR.18.4.657-686.2005, PMID 16223952, PMCID PMC1265908.

  3. Pitout JD, Laupland KB. Extended spectrum beta lactamase producing enterobacteriaceae: an emerging public health concern. Lancet Infect Dis. 2008 Mar;8(3):159-66. doi: 10.1016/S1473-3099(08)70041-0, PMID 18291338.

  4. Steward CD, Wallace D, Hubert SK, Lawton R, Fridkin SK, Gaynes RP. Ability of laboratories to detect emerging antimicrobial resistance in nosocomial pathogens: a survey of project ICARE laboratories. Diagn Microbiol Infect Dis. 2000 Sep;38(1):59-67. doi: 10.1016/s0732-8893(00)00161-9, PMID 11025185.

  5. Gupta V. An update on newer beta lactamases. Indian J Med Res. 2007 Nov;126(5):417-27. PMID 18160745.

  6. Livermore DM, Canton R, Gniadkowski M, Nordmann P, Rossolini GM, Arlet G. CTX-M: changing the face of ESBLs in Europe. J Antimicrob Chemother. 2007 Feb;59(2):165-74. doi: 10.1093/jac/dkl483, PMID 17158117.

  7. Sturenburg E, Mack D. Extended spectrum beta lactamases: implications for the clinical microbiology laboratory therapy and infection control. J Infect. 2003 Nov;47(4):273-95. doi: 10.1016/s0163-4453(03)00096-3, PMID 14556752.

  8. Perez F, Endimiani A, Hujer KM, Bonomo RA. The continuing challenge of ESBLs. Curr Opin Pharmacol. 2007 Oct;7(5):459-69. doi: 10.1016/j.coph.2007.08.003, PMID 17875405, PMCID PMC2235939.

  9. Romero ED, Padilla TP, Hernandez AH, Grande RP, Vazquez MF, Garcia IG. Prevalence of clinical isolates of escherichia coli and klebsiella spp. producing multiple extended-spectrum beta-lactamases. Diagn Microbiol Infect Dis. 2007 Dec;59(4):433-7. doi: 10.1016/j.diagmicrobio.2007.06.007, PMID 17913435.

  10. Kuo KC, Shen YH, Hwang KP. Clinical implications and risk factors of extended spectrum beta lactamase producing klebsiella pneumoniae infection in children: a case control retrospective study in a medical center in southern Taiwan. J Microbiol Immunol Infect. 2007 Jun;40(3):248-54. PMID 17639166.

  11. Hosoglu S, Gundes S, Kolayli F, Karadenizli A, Demirdag K, Gunaydin M. Extended spectrum beta lactamases in ceftazidime resistant escherichia coli and klebsiella pneumoniae isolates in Turkish hospitals. Indian J Med Microbiol. 2007 Oct;25(4):346-50. doi: 10.4103/0255-0857.37336, PMID 18087082.

  12. Messai Y, Iabadene H, Benhassine T, Alouache S, Tazir M, Gautier V. Prevalence and characterization of extended spectrum β-lactamases in klebsiella pneumoniae in Algiers hospitals (Algeria). Pathol Biol (Paris). 2008;56(5):319-25. doi: 10.1016/j.patbio.2008.05.008, PMID 18585867.

  13. Jacoby GA, Munoz Price LS. The new beta lactamases. N Engl J Med. 2005 Jan 27;352(4):380-91. doi: 10.1056/NEJMra041359, PMID 15673804.

  14. Clinical Laboratory Standards Institute. CLSI performance standards for antimicrobial susceptibility testing. In: proceedings of the 16th international supplement (M100-S16) national committee for clinical laboratory standards, wayne PA USA; 2006.

  15. Murugan N, Malathi J, Therese KL, Madhavan HN. Antimicrobial susceptibility and prevalence of extended spectrum betalactamase (ESBL) and metallo-beta-lactamase (MBL) and its co-existence among pseudomonas aeruginosa recovered from ocular infections. Int J Pharm Pharm Sci. 2015 May 1;7(5):147-51.

  16. National Committee for Clinical Laboratory Standards. NCCLS. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, approved standard M7-A5 and informational supplement M100-S10. Wayne PA: National Committee for Clinical Laboratory Standards; 2000.

  17. Shashwati N, Kiran T, Dhanvijay AG. Study of extended spectrum β-lactamase producing enterobacteriaceae and antibiotic coresistance in a tertiary care teaching hospital. J Nat Sci Biol Med. 2014 Jan;5(1):30-5. doi: 10.4103/0976-9668.127280, PMID 24678193, PMCID PMC3961948.

  18. Mathur P, Kapil A, Das B, Dhawan B. Prevalence of extended spectrum beta lactamase producing gram negative bacteria in a tertiary care hospital. Indian J Med Res. 2002 Apr;115:153-7. PMID 12239838.

  19. Hansotia JB, Agarwal V, Pathak AA, Saoji AM. Extended spectrum beta lactamase-mediated resistance to third-generation cephalosporins in klebsiella pneumoniae in Nagpur central India. Indian J Med Res. 1997 Apr;105:158-61. PMID 9145597.

  20. Lal P, Kapil A, Das BK, Sood S. Occurrence of TEM and SHV gene in extended spectrum beta-lactamases (ESBLs) producing klebsiella sp. isolated from a tertiary care hospital. Indian J Med Res. 2007 Feb;125(2):173-8. PMID 17431288.

  21. Rodrigues C, Joshi P, Jani SH, Alphonse M, Radhakrishnan R, Mehta A. Detection of lactamases in nosocomial gram negative clinical isolates. Indian J Med Microbiol. 2004 Oct-Dec;22(4):247-50. doi: 10.1016/S0255-0857(21)02772-9, PMID 17642748.

  22. Ananthan S, Subha A. Cefoxitin resistance mediated by loss of a porin in clinical strains of klebsiella pneumoniae and escherichia coli. Indian J Med Microbiol. 2005 Jan;23(1):20-3. doi: 10.4103/0255-0857.13867, PMID 15928416.

  23. Gupta S, Maheshwari V. Prevalence of ESBLs among enterobacteriaceae and their antibiotic resistance pattern from various clinical samples. Int J Curr Microbiol App Sci. 2017;6(8):2620-8. doi: 10.20546/ijcmas.2017.609.323.

  24. Mendelson G, Hait V, Ben Israel J, Gronich D, Granot E, Raz R. Prevalence and risk factors of extended spectrum beta lactamase producing escherichia coli and klebsiella pneumoniae in an Israeli long-term care facility. Eur J Clin Microbiol Infect Dis. 2005 Jan;24(1):17-22. doi: 10.1007/s10096-004-1264-8, PMID 15660255.

  25. Bazzaz BS, Naderinasab M, Mohamadpoor AH, Farshadzadeh Z, Ahmadi S, Yousefi F. The prevalence of extended-spectrum beta-lactamase-producing escherichia coli and klebsiella pneumoniae among clinical isolates from a general hospital in Iran. Acta Microbiol Immunol Hung. 2009 Mar;56(1):89-99. doi: 10.1556/AMicr.56.2009.1.7, PMID 19388560.

  26. Xiong Z, Zhu D, Zhang Y, Wang F. Extended spectrum beta lactamase in klebsiella pneumoniae and escherichia coli isolates. Zhonghua Yi Xue Za Zhi. 2002 Nov 10;82(21):1476-9. PMID 12509910.

  27. Goossens H, MYSTIC Study Group (Europe). MYSTIC program: summary of European data from 1997 to 2000. Diagn Microbiol Infect Dis. 2001 Dec;41(4):183-9. doi: 10.1016/s0732-8893(01)00320-0, PMID 11777657.

  28. Babypadmini S, Appalaraju B. Extended spectrum lactamases in urinary isolates of escherichia coli and klebsiella pneumonia: prevalence and susceptibility pattern in a tertiary care hospital. Indian J Med Microbiol. 2004 Jul-Sep;22(3):172-4. doi: 10.1016/S0255-0857(21)02830-9, PMID 17642726.