Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital

Dr. R. Suganya et al. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital. Int. J Med. Pharm. Res., 6 (6): 622‐628, 2025 622 International Journal of Medical and Pharmaceutical Research Online ISSN-2958-3683 | Print ISSN-2958-3675 Frequency: Bi-Monthly Available online on: https://ijmpr.in/ Original Article Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital Dr. N. Subathra1, Dr. R. Suganya2, Dr. S.K Jayaswarya3 1 MD, Associate Professor of Microbiology Govt Medical College Namakkal. 2 MD, Assistant Professor of Microbiology GMKMC Salem. 3 Assistant professor of Microbiology Govt Medical College Namakkal. A B S T R A C T Corresponding Author: Dr. R. Suganya MD, Assistant Professor of Microbiology GMKMC Salem. Received: 10-10-2025 Accepted: 14-11-2025 Available online: 22-11-2025 Background: Body fluids such as cerebrospinal, pleural, peritoneal, pericardial, and synovial fluids are normally sterile. Infections of these sites lead to significant morbidity and mortality. Early bacteriological identification and antimicrobial susceptibility profiling are essential for effective patient management. Aim: To determine the bacteriological profile and antimicrobial susceptibility pattern of isolates from sterile body fluids at a tertiary-care hospital. Materials and Methods: This retrospective laboratory-based observational study was conducted in the Department of Microbiology, Government Mohan Kumaramangalam Medical College Hospital (GMKMCH), Salem, Tamil Nadu, from June 2020 to March 2021. Four hundred sixty-five body-fluid samples were processed by standard microbiological procedures. Antimicrobial susceptibility testing was performed by the Kirby–Bauer di sk-diffusion method and interpreted as per CLSI 2020 guidelines. Results: Of 465 samples, 99 (21.3 %) showed growth. Pseudomonas aeruginosa (24.2 %) was the most common isolate, followed by Klebsiella spp. (23.2 %), Acinetobacter spp. (17.2 %), and Staphylococcus aureus (17.2 %). MRSA prevalence was 23.5 %. Gram-negative isolates exhibited highest sensitivity to carbapenems and piperacillin–tazobactam, while Gram-positives were 100 % sensitive to linezolid and vancomycin. Conclusion: Gram-negative bacilli predominated among isolates from sterile body fluids. Continuous surveillance of local antimicrobial trends is vital for guiding empirical therapy and antibiotic-stewardship policies. Copyright © International Journal of Medical and Pharmaceutical Research Keywords: Sterile body fluids, bacteriological profile, antimicrobial susceptibility, MRSA, gram negative. INTRODUCTION Body fluids play a substantial role in transporting nutrients, regulating body temperature, and aiding respiration (Abdinia et al.). Normally sterile fluids such as cerebrospinal, pleural, peritoneal, synovial, and pericardial fluid can become infected by microorganisms—bacteria, fungi, viruses, or parasites—leading to severe morbidity and mortality (Deb et al.; Hasbun et al.). Early detection and identification of pathogens are essential for proper management and reduced hospital stay (Sujatha et al.). Common pathogens include Escherichia coli, Klebsiella spp., Haemophilus influenzae, Staphylococcus aureus, Neisseria meningitidis, Pseudomonas spp., and Acinetobacter spp. Because these infections are medical emergencies, regular monitoring of local antibiograms is essential for effective empirical therapy and public-health antibiotic policies. MATERIALS AND METHODS Study Design and Duration: This cross-sectional study was conducted in the Department of Microbiology, Government Mohan Kumaramangalam Medical College Hospital (GMKMCH), Salem, Tamilnadu from June 2020 to April 2021. Sample Collection: A total of 465 sterile body fluid samples including cerebrospinal, pleural, peritoneal, synovial, and pericardial fluids were collected using strict aseptic precautions and transported within two hours. Culture and Identification: Samples were inoculated on Blood agar, MacConkey agar, and Chocolate agar plates and incubated at Dr. R. Suganya et al. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital. Int. J Med. Pharm. Res., 6 (6): 622‐628, 2025 623 37°C for 24–48 hours. Bacterial identification was performed using Gram staining, colony morphology, and standard biochemical reactions such as oxidase, indole, citrate, urease and triple sugar iron test. Antimicrobial Susceptibility: Antibiotic sensitivity testing was carried out using the Kirby–Bauer disk diffusion technique on Mueller–Hinton agar as per CLSI 2020 guidelines. Quality Control: Reference ATCC strains—E. coli 25922, Staphylococcus aureus 25923, and Pseudomonas aeruginosa 27853 were employed as ATCC. Data Analysis: Data were analyzed using Microsoft Excel 2021 and presented as percentages. Ethics: Institutional Ethical Committee approval was obtained prior to commencement of the study. Ethical clearance was obtained from the Institutional Ethics Committee. Inclusion criteria included all body-fluid samples received from admitted patients with suspected infection, irrespective of age or sex. Exclusion criteria excluded blood samples, patients with recent antibiotic therapy (within two weeks), contaminated samples, and samples delayed beyond two hours after collection. Each specimen was examined microscopically (Gram stain) and cultured on blood agar, MacConkey agar, and chocolate agar. Bacterial identification was performed by stan biochemical tests (Collee et al.). Susceptibility testing was performed using the Kirby–Bauer disk-diffusion method on Mueller-Hinton agar and interpreted as per CLSI 2020 guidelines. Out of 465 processed samples, 99 (21.3%) exhibited bacterial growth. Gram-negative organisms predominated, particularly Pseudomonas aeruginosa, Klebsiella species, Acinetobacter species, and Escherichia coli. Gram-positive isolates included Staphylococcus aureus and coagulase-negative Staphylococci (CONS). Lactose-fermenting isolates demonstrated excellent sensitivity to β-lactam/β-lactamase inhibitor combinations and carbapenems, whereas non-fermenting isolates exhibited varying resistance patterns. Table 1 & Figure 1: Growth pattern of body fluids Sample type Total number of samples Growth (%) No Growth (%) Pleural fluid 218 45 (21%) 173 (79%) Ascitic fluid 146 41(28%) 105 (72%) Cerebrospinal fluid 79 08 (10%) 71 (90%) Synovial fluid 12 04 (33%) 08 (67%) Pericardial fluid 08 00 (00%) 08 (100%) Bile 02 01 (50%) 01 (50%) Total 465 99 (21%) 366 (79%) Table 2 & Figure 2: Bacterialogical profile of different body fluid samples 21% 28% 10% 33% 0% 50% Pleural fluid Ascitic fluid Cerebrospinal fluid Synovial fluid Pericardial fluid bile Organisms Total 465(99) Pleural fluid 218 (45) Ascitic fluid 146 (41) Cerebrospinal fluid 79 (08) Synovial fluid 12 (04) Pericardial fluid 08 (00) Bile 02 (01) Klebsiella spp 23 09 11 02 - - 01 E.coli 08 01 05 02 - - - Pseudomonas spp 24 16 08 - - - - Dr. R. Suganya et al. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital. Int. J Med. Pharm. Res., 6 (6): 622‐628, 2025 624 Table 3: Antibiotic susceptibility pattern of Gram negative bacteria (GNB). (N = 33) Antibiotics Klebsiellaspp n=23 E. coli n = 8 Citrobacterspp n = 2 Ampicillin 5% 7% 18% Amoxycillin clavulanic acid 42% 47% 62% Amikacin 74% 85% 92% Gentamicin 72% 84% 90% Ciprofloxacin 26% 27% 33% Ceftriaxone 33% 35% 46% Cefotaxime 36% 34% 48% Piperacillin Tazobactam 98% 100% 100% Cefoperazonesulbactam 96% 98% 100% Imepenem 100% 100% 100% Meropenem 100% 100% 100% Cotrimoxazole 33% 50% 70% Doxycycline 96% 100% 100% Acinetobacter spp 17 09 05 03 - - - Citrobacter spp 02 02 - - - - - Staph aureus 17 06 06 01 04 - - CONS 07 02 05 - - - - Streptococcus spp 01 - 01 - - - - Dr. R. Suganya et al. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital. Int. J Med. Pharm. Res., 6 (6): 622‐628, 2025 625 Table 4: Antibiotic susceptibility pattern of Non Fermenting Gram negative bacteria (NFGNB). (N = 41) Antibiotics Pseudomonas spp n = 24 Acinetobacterspp n = 17 Ampicillin ND 5% Amoxycillin clavulanic acid ND 30% Amikacin 96% 94% Gentamicin 83% 76% Ciprofloxacin 50% 53% Ceftriaxone ND 32% Cefotaxime ND 32% Ceftazidime 28% 30% Piperacillin Tazobactam 92% 88% Cefoperazonesulbactam 88% 82% 0% 20% 40% 60% 80% 100% 120% Ampicillin Amoxycillin clavulanic acid Amikacin Gentamicin Ciprofloxacin Ceftriaxone Cefotaxime Piperacillin Tazobactam Cefoperazonesulbactam Imepenem Meropenem Cotrimoxazole Doxycycline Chart Title Table 3: Antibiotic susceptibility pattern of Gram negative bacteria (GNB). (N = 33) Citrobacterspp n = 2 Table 3: Antibiotic susceptibility pattern of Gram negative bacteria (GNB). (N = 33) E.coli n = 8 Table 3: Antibiotic susceptibility pattern of Gram negative bacteria (GNB). (N = 33) Klebsiellaspp n=23 Dr. R. Suganya et al. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital. Int. J Med. Pharm. Res., 6 (6): 622‐628, 2025 626 Imepenem 96% 96% Meropenem 94% 94% Cotrimoxazole ND 62% Table 5 & Figure 5: Antibiotic susceptibility pattern of Gram-positive isolates (n – 25) Antibiotics Staphylococcus aureus (n = 17) CONS (n = 7) Streptococcus spp (n = 1) Cefoxitin 76% 100% ND Gentamicin 59% 86% 100% Ciprofloxacin 65% 29% 100% Cotrimaxazole 24% 29% 100% Doxycycline 41% 43% 100% Erythromycin 12% 14% 100% Clindamycin 18% 14% 100% Vancomycin 100% 100% 100% Linezolid 100% 100% 100% Ampicillin - - 100% Amoxyclav - - 100% Cefotaxime - - 100% 0 0.5 1 1.5 2 2.5 Chart Title Table 4:Antibiotic susceptibility pattern of Non Fermenting Gram negative bacteria (NFGNB). (N = 41) Acinetobacterspp n = 17 Table 4:Antibiotic susceptibility pattern of Non Fermenting Gram negative bacteria (NFGNB). (N = 41) Pseudomonas spp n = 24 Dr. R. Suganya et al. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital. Int. J Med. Pharm. Res., 6 (6): 622‐628, 2025 627 DISCUSSION The overall culture positivity rate of 21.3 % was consistent with studies by Sharma et al., Harshika et al., and Vishalakshi et al., who reported positivity between 20–30 %. Pseudomonas aeruginosa was the predominant isolate, aligning with findings of Harshika et al. and Sharma et al. Klebsiella spp. and Acinetobacter spp. were next most frequent, similar to reports by Vishalakshi et al. and Abdinia et al. S. aureus and CONS isolation patterns correlated with Sujatha et al. and Deb et al. All Gram-positive isolates remained 100 % sensitive to vancomycin and linezolid. Carbapenems and piperacillintazobactam retained the highest activity among Gram-negative isolates. These trends emphasize the need for ongoing surveillance to guide empirical antimicrobial therapy. CONCLUSION Gram-negative bacteria remain the predominant pathogen among sterile body-fluid isolates, with Pseudomonas aeruginosa being the most common organism. Carbapenems and β-lactam/β-lactamase inhibitor combinations remain effective empirical treatment options while resistance to cephalosporins and fluroquinolones is rising. Continuous surveillance of local bacteriological profiles and antimicrobial patterns is essential for rational antibiotic therapy and stewardship Ethical Clearance Obtained from Institutional Ethics Committee, GMKMCH, Salem. Acknowledgement The authors thank the Department of Microbiology, GMKMCH, Salem, for technical support. REFERENCES 1. Abdinia B et al. Epidemiology and bacterial profile of sterile body fluid infections. J Clin Diagn Res. 2014;8(5): DC20–DC23. 2. Sharma R, Anuradha, Nandini D. Bacteriological Profile and Antimicrobial Sensitivity pattern in Sterile Body Fluids from a Tertiary Care Hospital. J Appl Microbiol Biochem. 2017, 1:1 3. Harshika Y K1, ShobhaMedegar K. R2, *, Asha B Patil3, Smita N R4A study on bacteriological profile and antimicrobial resistance pattern from various body fluids of patients attending the tertiary care Hospital, KIMS, Hubli 4. A Study on Aerobic Bacteriological Profile of Sterile Body Fluids B. Vishalakshi*, Pushpalatha Hanumanthappa and S. Krishna 5. Bacterial Isolates and Drug Susceptibility Pattern of Sterile Body Fluids from Tertiary Hospital, Northern Ethiopia: AFour-Year Retrospective Study EphremTsegay,1 AregawiHailesilassie,2 Haftamu Hailekiros ,1 SelamNiguse,1 Muthupandian Saravanan ,1andMahmudAbdulkade 6. Deb AK et al. Bacteriological profile and antibiotic sensitivity pattern of isolates from sterile body fluids. Indian J Med Microbiol. 2014;32(4):508–513. 7. Hasbun R et al. Bacterial infections in body fluids: clinical outcomes and management. Clin Infect Dis. 2013;56(9):1251–1258. 8. Sujatha R et al. Microbial profile and antimicrobial susceptibility from body fluids in tertiary care hospitals. J Lab Physicians. 2015;7(2):95–100. 9. Wiest R et al. Infections in cirrhosis and ascitic fluid: pathophysiology and treatment. J Hepatol. 2011;55(5):1228– 1241. 10. Van de Beek D et al. Bacterial meningitis in adults. Lancet. 2012;380(9854):1703–1712. 11. World Health Organization. Global Antimicrobial Resistance and Use Surveillance System (GLASS) Report. Geneva: WHO; 2020. 0 0.2 0.4 0.6 0.8 1 1.2 010 20 30 40 Table 5:Antibiotic susceptibility pattern of Gram positive isolates (n – 25) Antibiotics Cefoxitin Table 5:Antibiotic susceptibility pattern of Gram positive isolates (n – 25) Staphylococcus aureus (n = 17) 76% Dr. R. Suganya et al. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Isolates from Sterile Body Fluids in a Tertiary Care Hospital. Int. J Med. Pharm. Res., 6 (6): 622‐628, 2025 628 12. M. Gizachew, H. Abdella, and M. Tiruneh, “Antimicrobial susceptibility patterns of staphylococcus aureus at the university of gondar tertiary hospital, northwest ethiopia: a retrospective cross-sectional study,” Journal of Bacteriology & Parasitology, vol.6, no.3, article 228,2015. 13. Jain Sonali, Banavaliker J N, Empyema Thoracis: Bacteriological analysis of pleural fluid, IOSR-JDMS. 2013;3(6):4651. ISSN: 2279-0853, ISBN:2279-0861. 14. Chen CJ, Huang YC (2005) Community-acquired methicillin resistant Staphylococcus aureus in Taiwan. MicrobiolImmunol Infect 38: 376-382