BACTERIAL PROFILE AND ANTIBIOGRAM OF BODY FLUIDS ISOLATED FROM VARIOUS IPDS PATIENTS

BACTERIAL PROFILE AND ANTIBIOGRAM OF BODY FLUIDS ISOLATED FROM VARIOUS IPDS PATIENTS

Mahesh Chandra * and Ansar Ahmad Paray

Department of Microbiology, Adesh Institute of Medical Science & Research Bathinda, Punjab, India.

ABSTRACT: Introduction: Pathogenic infections place a significant burden on the healthcare system since they are linked to high rates of morbidity and mortality. Successful therapy of these illnesses depends on an early diagnosis and a good antimicrobial resistance testing. Therefore, the goal of the current investigation was to assess aerobic bacteriological profile and antibiogram from a variety of sterile fluids. Material and Method: This observational study was carried out in the Department of Microbiology, Adesh Institute of Medical Science & Research Bathinda, Punjab from 20 September 2023 to 15 February 2024 obtaining the approval from Ethics Committee of Biomedical & Health Research, Adesh University. All the Body fluid samples were received in Bacteriology Laboratory from various departments. All body fluid samples are processed by Bact alert system. Positive samples were further subculture on blood agar and MacConkey agar. Gram Staining was done to identify gram-positive bacteria and gram-negative Bactria, further antimicrobial susceptibility was done by Vitek 2 compact system as per CLSI guidelines 2023. Results: A total of 288 body fluid samples were received, from which Pus & Swab 102 (38.05%) were highest in number. Among the 157 isolates, the most predominant organism was Escherichia Coli 37 (23.56%) followed by Staphylococcus aureus 29 (18.47%), Klebsiella pneumoniae. The most sensitive drugs for gram-positive isolates were Daptomycin, Teicoplanin, Vancomycin, Doxycycline, Minocycline and for gram negative isolates were Amikacin, Minocycline, Tigecycline, Fosfomycin. Conclusion: The study concluded that multidrug-resistant bacteria are mostly responsible for infections of sterile body fluids and their antibiotic resistance varies greatly across various geographic areas and healthcare settings. Therefore, empirical treatment and antimicrobial testing crucial for effective treatment.

Keywords: Antimicrobial resistance, Gram-positive bacteria, Gram-negative bacteria, Klebsiella pneumoniae, Staphylococcus aureus

INTRODUCTION: Pathogenic infections place a significant burden on the healthcare system since they are linked to high rates of morbidity and mortality ¹. With sterile bodily fluids (SBF), there are no bacteria that would typically be present in the flora. Microorganisms, however, can enter sterile bodily sites ^{2, 3}.

Leading to significant invasive illnesses and catastrophic phenomena ^{4, 5}. Fluids may build up in any body cavity because of infection, leading to invasive disorders such Bacteremia, bacterial meningitis, sepsis, bacterial peritonitis, and other problems ⁶.

Because un-treated infections at sterile body sites can result in severe, potentially fatal infections elsewhere in the body, these infections are critical, urgent conditions that need to be treated right once ⁷. Particularly, infections brought on by resistant microorganisms continue to be a major global cause of severe infections, with rising rates of morbidity and mortality ^{8, 9}.

It is largely more common in developing nations where there are fewer health care services, fewer resources, inadequate sanitation and hygiene, irrational antibiotic usage, and poverty are some of the main factors contributing to the formation of antibiotic-resistant microorganisms ^{10, 11}. The present study aim was to find the prevalence of bacteremia from body fluid samples and performed their antibiotic sensitivity pattern.

MATERIAL AND METHOD: The study was carried out in the Department of Microbiology, Adesh Institute of Medical Science & Research Bathinda, Punjab from 20 September 2023 to 15 February 2024 after obtaining the approval from Ethics Committee of Biomedical & Health Research, Adesh University (reference no. AU/EC_BHR/2K23/486). Body fluid and swab samples were received from various departments. The sample size was calculated by Daniel’s formula with 5% of margin error ¹². Sample size was 288 body fluid samples such as ascitic fluid, pleural fluid, cerebral fluid, pericardial fluid, bronchial wash fluid, drain fluid, Synovial fluid, and pus samples. The sample size was calculated using the following formula calculated using the following formula:

n = Z x P x q / e²

(1.96)² x (0.25) x (0.75) / (0.05)² = 0.72 / 0.0025 = 288

n = Sample size, Z = the standard deviate, usually 1.96 which corresponds to 95% confidence P = Expected average Prevalence. According to Previous studies (25%), e = Margin of error (0.05).

The sample received in the laboratory in proper conditions was immediately processed for fluid culture by Bact alert systems, the positive samples as indicated by Bact alert systems were sub-cultured on blood agar and MacConkey agar. Swab/pus samples were directly inoculated on Blood agar and MacConkey agar and incubated at 37°C.

Identification of the Bacterial Isolates: After bact alert system indicating growth of the organism, Subculture was done on Blood agar and MacConkey agar from the Bact alert culture bottles. Swab/Pus samples were directly inoculated on blood agar and MacConkey agar using directly streak culture method. The culture plate were incubated at 37 °C for 18-24 hours in the incubator. Primary identification was done on the basis of colony characteristics, gram- staining and gram staining followed by final identification using Vitek 2 compact automated systems using Gram negative (GN) cards for gram negative and Gram positive (GP) cards for gram positive isolates.

Antimicrobial Susceptibility Test (AST): AST was performed by automated method using Vitek 2 compact system using AST cards P628 for gram positive bacteria and N405 & N406 for gram-negative bacteria as per Clinical Laboratory standard Institute (CLSI) guidelines 2023 ¹³. Statical Analysis was done by using Microsoft world and Microsoft excel software.

RESULTS: During the study period between 20 September 2023 to 15 February 2024 total 288 randomly non-repetitive body fluid and Swab/pus samples obtained from various departments of hospital. Culture positivity was seen in 157 (54.51%) samples and 131 (45.48%) samples showed no growth.

Out of the 157 samples, 106 (67.50%) were of males and 51 (32.48%) females. The positive samples belong to maximum from age group 61-70 years as compared to other age groups Fig. 1. Out of 157 positive cultures 123 (78.34%) were gram negative bacteria and 34 (21.66 %) gram positive bacteria. Among the 157 isolates, the most predominant organism was Escherichia Coli 37 (23.56%) followed by Staphylococcus aureus 29 (18.47%), Klebsiella pneumonia (15.9%) as shown in Fig. 2.

TABLE 1: SAMPLES WISE DISTRIBUTION OF BACTERIAL PREVENANCE

FIG. 1: AGE WISE DISTRIBUTION OF BACTERIAL SPECIES                                    

FIG. 2: DISTRIBUTION OF BACTERIAL ISOLATED

In the present study, S. aureus show (100%) sensitivity to Daptomycin, Teicoplanin, Vancomycin, Doxycycline, Minocycline followed by Rifampicin (96.50%), Linezolid (96.03 %), Tetracycline (95.00%) Gentamicin (79.40%) & Erythromycin (51.85%). E. coli shows (86.15%) sensitivity to Fosfomycin and (75.65%) to Amikacin followed by Ertapenem (68.40%), Imipenem (56.70%), Meropenem (54.00%), Cefoperazone / Sulbactam (51.35%) & Minocycline (50.00%). Klebsiella pneumoniae shows highly sensitive for Amikacin (92.00%) followed by Tigecycline (60.00%), Gentamicin (48.00%), Levofloxacin & Minocycline (42.80%). The antibiotic sensitivity patterns result of gram-positive cocci are show in Table 2 and those are gram negative show in Table 3.

TABLE 2: ANTIMICROBIAL SENSITIVITY (%) PATTERN OF GRAM-POSITIVE COCCI

NT- Not tested.

TABLE 3: ANTIMICROBIAL SENSITIVITY (%) PATTERN OF GRAM-NEGATIVE BACILLI

NT- Not tested.

DISCUSSION: Normally body fluids like Cerebral spinal, ascitic, pleural, cerebral, pericardial, synovial fluids and pus samples are usually sterile but can be infected by various pathogens. In the study the isolation rate of body fluid showing growth positive was 157 (54.51%) which correlates with studies conducted by Duggal et al.,¹⁴, Mita et al.,¹⁵ and Deboral et al.,¹⁶. They had also reported culture positivity rate of 93%, 85% and 72% respectively. The low rate of isolation may be explained by the fact of the patient probably received antibiotic therapy before culture sensitivity testing.

In the present study the maximum bacterial isolates were isolated from Pus/ swab (83.63%) samples as followed by Drain (53.4%), Bal (Bronchial wash) (52.38%), Synovial samples (20.00%), Pleural (22.22%), CSF (13.33%) and Ascitic (10.00%). Studies done by Shume et al., ¹⁷ and Singh et al.,¹⁸ had also reported prevalence from Pleural fluid (29.8%), (14%) respectively. Sadhna et at., had reported prevalence from CSF (34.4%), drain fluid (13.11%), BAL fluid (6.55%), ascitic fluid (4.91%), pleural fluid (3.27%), synovial fluid, (1.63%) ¹⁹. The results of their studies correlated with this present study. Whereas studies by Shume et al.,¹⁷ and Shrestha et al., ²⁰ reported low rates of prevalence from body fluid culture accounting for 16.7% and 10.68% respectively.

The present study showed males patients had high culture positivity as compared with females’ patients. i.e. 106 (67.50%) males and 51 (32.50%) were females. The result was compared with the study done by Singh et al., ¹⁹ and Shume et al., ¹⁷ they are showing 57.30%, 63.70% in males and 42.70%, 32.40% in females respectively. The reason for some difference is because of gender Bais or high admission rate of males.

In the present study, the isolation of gram-negative bacteria (78.30%) is higher than gram positive bacteria (21.70%). Which is consistent with the previous studies conducted by Shume et al., and Madigubba et al., showing gram-negative bacteria 70.60%, 83.20%. and Gram-positive bacteria 29.40%, 16.35% respectively ^{17, 21}. Another similar study done by Rouf et al., shows Gram-negative bacteria 70.00% as compared to gram positive bacteria (30%) ²².

In our study, the most Common organisms were Escherichia Coli 37 (23.5%) followed by Staphylococcus aureus 29 (18.55%), Klebsiella pneumoniae (15.95%), Pseudomonas aeruginosa (15.90%), Enterobacter cloacae complex (7.64%), Acinetobacter baumannii (6.36%), Proteus mirabilia (5.10%), Serratia marcescens (2.60%), Sphingomonas paucimobilis (1.91%), CoNS (1.91%), Enterococcus faecium (1.27%).This finding correlates  with study done by  Madigubba et al., study, showing most Common organisms were Escherichia coli (29.80%), Klebsiella pneumoniae (14.70%), Enterobacter spp. (5.00%), Acinetobacter baumannii (13.7%), Pseudomonas aeruginosa (13.10%), Staphylococcus aureus (4.50%) ²¹. Another similar study done by Harshika et al., they are showing predominant organism isolated was Escherichia coli (23.2%), followed by Pseudomonas (14%), Klebsiella (13.4%), Staphylococcus aureus (10.50%). Less commonly isolated were Enterococcus species (2.00%), Enterobacter (1.40%), and Streptococcus pyogenes (0.70%) ²³. One more study done by Rouf and Nazir, showing E. coli was most in gram-negative organism ²².

In the present study, antimicrobial susceptibility pattern among the Gram‑negative organisms showed that E. coli was highly sensitive to Fosfomycin (86.15%) and Amikacin (75.65%) followed by Ertapenem (68.40%), Imipenem (56.70%), Meropenem (54.00%), Cefoperazone / Sulbactam (51.35%) & Minocycline (50.00%). Similar study done by Madigubba, et al., ²¹ they are showing highly sensitive to amikacin (83.00%), meropenem (81.00%), Cefoperazone‑sulbactam (62.70%) and piperacillin‑tazobactam (57.30%). Other antibiotics show similar sensitive to Ceftazidime and Ceftriaxone (20.80%), (16.10%) respectively. E. coli showing less sensitive to Gentamicin (25.00%). Another study done by Harshika et al., they are showing highly sensitive to Amoxyclav, Ciprofloxacin (44.00%), (62.00%) respectively. Amikacin showed highly resistance (38.00%) ²³.

In the present study, Klebsiella pneumoniae shows sensitive for Amikacin (92.00%) followed by Tigecycline (60.00%), Gentamicin (48.00%), Levofloxacin & Minocycline (42.85%). According to another study by Harshika et al., ²³ showing Gentamicin (78.45%), Amoxyclav (44.00%), Ceftriaxone (38.00%) and Cefotaxime (34.00%). They are showing less sensitive to Amikacin (84.00%). One more study done by Madigubba et al., showing lesser sensitive to Amikacin (59.80%), Gentamicin (21.80%). Ciprofloxacin, Ceftazidime, Ceftriaxone were lesser resistant (65.50%), (72.40%), (74.50%) respectively ²¹.

In the present study, S. aureus showed (100%) sensitivity to Daptomycin, Teicoplanin, Vancomycin, Doxycycline, Minocycline followed by Rifampicin (96.50%), Linezolid (96.50 %), Tetracycline (95.00%), Gentamicin (79.40%) & Erythromycin (51.80%). The present study corallites with Rouf and Nazir, they are showing were highly sensitive to Vancomycin, Linezolid (100%), Tigecycline (60%-70%), Teicoplanin (50%-60%) ²². Similar study done by Harshika et al., showing 100% sensitive to Amikacin, Gentamicin, Vancomycin, followed by Ciprofloxacin, Levofloxacin, Cefepime (92.00%), Amoxyclav (84.00%), Clindamycin (75.00%), Erythromycin (67.00%), Tetracycline (56.00%) ²³.

CONCLUSION: In the present study Escherichia coli, followed by Staphylococcus aureus and Klebsiella pneumonia were the predominant isolates from various body fluids. Most of the Gram-positive isolates were resistant to Benzylpenicillin, Ciprofloxacin, Levofloxacin Oxacillin. Gram negative isolates were highly resistant to Amoxicillin/Clavulanic Acid, Cefuroxime, Cefuroxime Axetil, Cefotaxime, Ceftriaxone. Moreover, the rates of multiple drug-resistant isolates are alarmingly. Therefore, the study recommends hospitals to have strict antibiotics utilization policies and to support clinicians for rational choice of antibiotic therapy and to be updated with the present antimicrobial patterns of locality.

ACKNOWLEDGEMENT: Nil

Funding/Grant declared: Nil

Authors’ Contribution: All authors contributed equally to the study.

Ethical Considerations: Ethical issues (including plagiarism, data fabrication, double publication) have been completely observed by authors.

CONFLICT OF INTEREST: Nil

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    How to cite this article:

    Chandra M and Paray AA: Bacterial profile and antibiogram of body fluids isolated from various IPDS patients. Int J Pharm Sci & Res 2024; 15(10): 3084-89. doi: 10.13040/IJPSR.0975-8232.15(10).3084-89.

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