Original article / research
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Bacteriological Profile and Antimicrobial Susceptibility Pattern of the Isolates from Body Fluid Samples from Tertiary Care Health Centre, Ujjain, Madhya Pradesh, India |
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Sonali Waske, Yogyata Marothi 1. Assistant Professor, Department of Microbiology, R.D. Gardi Medical College, Ujjain, Madhya Pradesh, India. 2. Professor and Head, Department of Microbiology, R.D. Gardi Medical College, Ujjain, Madhya Pradesh, India |
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Correspondence
Address : Sonali Waske, Assistant Professor, Department of Microbiology, RDGMC, Ujjain, Madhya Pradesh, India. E-mail: drsonalip@gmail.com |
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ABSTRACT | |||||||||||||||||||||||||||||||||||||||||||
: Bacterial Infection of various body fluids can leads to serious invasive infection, high morbidity and mortality. It is a clinical urgency; so early detection and identification of pathogen is essential for appropriate management of patient. Aim: To determine the bacterial profile and their susceptibility to antimicrobial agents of isolates from various body fluids. Materials and Methods: This retrospective cross-sectional study was conducted from January 2021 to December 2021 in the Microbiology Department, CRG Hospital, RD Gardi Medical College, Ujjain, Madhya Pradesh, India. Body fluids were collected and transported aseptically from the patients visited in this hospital. All received samples were processed as per standard microbiology methods. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method and interpreted as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: A total of 298 body fluids samples were received for culture and sensitivity, of these, 63 (21.1%) fluid samples showed growth. Out of them, maximum pathogens were isolated from Broncho Alveolar Lavage (BAL) fluid 28 (44.4%), followed by peritoneal fluid 17 (27%), pleural fluid 12 (19%), Cerebospinal Fluid (CSF) 1 (1.6%), pericardial fluid 1 (1.6%) and synovial 4 (6.3%). Most of the isolates were Gram-Negative Bacteria (GNB) 58 (92%), predominantly Pseudomonas aeruginosa 20 (32%) followed by Klebsiella species 18 (29%), E.coli 14 (22%), All Gram negative isolates were sensitive to Tigecycline and Colistin, However, susceptibility to the Carbapenem group (Imipenem, Miropenem and Ertapenem) were (80-100%). Among Gram positive bacteria, Staphylococcus aureus were 5 (8%) isolated. Of these, Methicillin Resistant Staphylococcus aureus (MRSA) was 4 (6.3%) and Methicillin Sensitive Staphylococcus Aureus (MSSA) was 1 (1.6%). All isolate of Staphylococcus aureus were susceptible to Vancomycin, Linezolide, Teicoplanin and Levofloxacin. Conclusion: Knowledge of bacteriological and antimicrobial profile of body fluids is necessary, so that the life threatening infections can be effectively treated. In this study, BAL fluid is the commonest received sample. GNB were main isolates among them Pseudomonas aeruginosa is frequently isolated pathogen. All isolated pathogens were susceptible for higher group of antimicrobial agents (tigecycline, colistin) and showed reduced sensitivity towards other antibiotics. So, regular monitoring of the pattern of Antimicrobial Resistance (AMR) of bacteriological isolates in the patients is critical to develop antibiotic policy to combat these infections and reduce morbidity and mortality. | |||||||||||||||||||||||||||||||||||||||||||
Keywords : Antimicrobial resistance, Bronchoalveolar lavage, Gram-negative bacteria, Gram-positive bacteria | |||||||||||||||||||||||||||||||||||||||||||
INTRODUCTION | |||||||||||||||||||||||||||||||||||||||||||
Sterile body sites are those in which no microbes exist as commensals when in a healthy state (1). Microorganisms can invade sterile body sites via haematogenous, lymphatic route or any means. Causative agents can be a pathogenic or opportunistic organism from the normal flora of body (2),(3). In response to infection, fluids may accumulate in any body cavity associated with invasive diseases, such as bacteraemia, sepsis, bacterial meningitis, bacterial peritonitis, and other complications (4). Infection of sterile Body fluids like BAL, ascitic (peritoneal fluid), pleural, pericardial, synovial fluids, cerebrospinal, and hydrocele are associated with grave morbidity and life threatening events (5). Lesser isolation rate of organisms from these samples is because of less number of pathogens as well as prior administration of empirical antibiotics (6). Hence, Isolation of single colony of microorganism is significant from body fluids (7),(8). Frequently isolated pathogenic organisms are Escherichia coli (E.coli), Pseudomonas aeruginosa (P.aeruginosa), Klebsiella pneumoniae, Staphylococcus aureus (S.aureus) and Non Fermenter Gram Negative Bacteria (NFGNB) (3). Emergence of AMR leads to extended hospital stay, higher medical costs, increases economical burden on the community, increased morbidity and mortality. The condition is depressing in the poor and developing countries because of limited healthcare facilities, lack of resources, poverty, low education, poor hygiene and sanitation, irrational and abrupt use of antibiotics (9). Early detection and rapid identification of microorganisms are crucial for the appropriate management in this situation (6). There are very limited data available on bacteriological profile and their antimicrobial susceptibility pattern from various body fluids samples from the central part of India (10),(11). However, this is a common topic and many studies conducted from the other parts of India (6),(12),(13),(14). So, the present study was undertaken to evaluate aerobic bacteriological profile along with their antibiogram that will be very useful to the clinicians for presumptive diagnosis and early start of the treatment. | |||||||||||||||||||||||||||||||||||||||||||
MATERIAL AND METHODS | |||||||||||||||||||||||||||||||||||||||||||
The retrospective cross-sectional study was conducted from January 2021 to December 2021 in the Department of Microbiology, CRG Hospital, RD Gardi Medical College, Ujjain, Madhya Pradesh, India, on 63 culture positive body fluids. Study Procedure A total of 298 sterile body fluids like pleural fluid, BAL fluid, peritoneal fluid, Cerebrospinal Fluid (CSF), synovial fluid etc., were collected aseptically from patients in a sterile container and transferred to the microbiology laboratory for further processing. With standard safety precautions, the samples were subjected for microscopic examination (Gram staining) and culture. Samples were processed by inoculation on blood agar, MacConkey’s agar and chocolate agar (Hi-media, Mumbai) and incubated aerobically at 37°C for 24 hours to obtain isolated colonies. Isolates were identified by using standard technique [7,8,15]. Isolates were identified on the basis of colony morphology, Gram staining, catalase test, coagulase test, indole, methyl reduction, urease, citrate, triple sugar iron test, mannitol motility test etc., [7,8]. Susceptibility of bacterial isolates to antimicrobial agents was carried out on Mueller Hinton Agar media using modified Kirby-Bauer disk diffusion method as per Clinical and Laboratory Standards Institute (CLSI) guideline (15). Relevant clinical history was collected from each patient for clinical correlation. Reference strains of S.aureus (ATCC 25923), E.coli (ATCC 25922), and P.aeruginosa (ATCC 27853) were used as quality control for susceptibility testing as per CLSI guideline (15). Statistical Analysis Descriptive statistical analysis were used to analyse data in form of number and percentage. | |||||||||||||||||||||||||||||||||||||||||||
RESULTS | |||||||||||||||||||||||||||||||||||||||||||
During the study period, a total of 298 body fluids were processed. Of these, 63 (21.1%) fluids showed growth. Among 63 culture positive samples, Majority were males 50 (79%) whereas females were 13 (21%). Age of the study group ranged from two years to 80 years with majority was above 50 years 31 (49%) (Table/Fig 1). The topmost sample was showed growth, that was BAL fluid 28 (44%), followed by peritoneal fluid 17 (27%), pleural fluid 12 (19%), CSF 1 (1.6%), pericardial fluid 1 (1.6%) and other body fluids 4 (6.3%) (synovial) (Table/Fig 2). Of the total 63 isolates, GNB were prevalent 58 (92%). Predominant GNB isolated was P.aeruginosa 20 (32%) followed by Klebsiella species 18 (29%), E.coli 14 (22%), Other NFGNB (Acinetobacter, Burkholderia) were 4 (6.3%) and less commonly isolated was Enterobacter species 2 (3.2%). S.aureus 5 (8%) was frequently isolated among Gram positive bacteria. MRSA were 4 (6.3%) and MSSA were 1 (1.6%) isolated (Table/Fig 2). All the S.aureus strains were sensitive to vancomycin, linezolide, teicoplanin and levofloxacin (Table/Fig 3). Gram negative organisms were utterly susceptible to tigecycline and colistin, sensitivity towards carbapenem group was 80-100%. They showed high resistance to amoxyclav, quinolones, aminoglycosides and third generation cephalosporin (ceftazidime, ceftriaxone and cefotaxime) (Table/Fig 4). | |||||||||||||||||||||||||||||||||||||||||||
DISCUSSION | |||||||||||||||||||||||||||||||||||||||||||
In the present study, samples were received from all the age groups, their age ranged from two year to 80 year, Majority of them were from above 50 years of the age with 79% males and 21% females. That is in concordance with study conducted at North India (63.6% were males, while 36.4% were females) (12). In the present study, culture positivity rate was 21.1% as it is likewise the study conducted by Harshika YK et al., with rate of 22% (13). In the present study, out of 63 culture positive samples, The most common received sample was BAL fluid 28 (44%), followed by peritoneal fluid, then pleural fluid, then CSF, pericardial fluid and other body fluids. This is in contrast with studies from South India, where peritoneal fluid and pleural fluid were reported as commonest received sample [6,13]. Isolation of microorganisms from various clinical samples was described in previous study in (Table/Fig 5) (6),(13),(14),(16),(17),(18). In this study P.aeruginosa 20 (32%) was the commonest pathogen isolated followed by Klebsiella species 18 (29%), E.coli 14 (22%), Other NFGNB were 4 (6.3%) and Enterobacter species 2 (3.2%) was least isolated among GNB. As compared to other studies, isolation rate of Gram positive bacteria were less, 5 (8%) were S.aureus, out of them MRSA were 4 (6.3%) and MSSA were 1 (1.1%). Bacteriological profile from various samples reported by past studies showed in (Table/Fig 6) (6),(13),(14),(16),(17),(18). In present study, GNB 58 (92%) were repeatedly isolated than Gram positive bacteria. This finding was similar to South Indian study with the isolation rate of Gram negative organisms were 83% and Gram positives were 16.3% (14). Similar Gram negative predominance seen in previous studies (3),(18). In contrast, Gram-positive bacteria was reported as most frequent isolate by Vishalakshi B et al., and Perween N et al., (6),(12). In the present study, predominant organism isolated was P.aeruginosa 20 (32%) followed by Klebsiella species 18 (29%), E.coli 14 (22%), That is correlated with previous studies by Shume T et al., and Magazine R et al., (3),(19). P.aeruginosa 15 (54%) was most commonly isolated from BAL fluid, that is comparable with studies conducted by Magazine R et al., 35 (35%) and Akhtar S et al., 46 (54.76%) (19),(20). Isolation rate of P.aeruginosa was 5 (26%) from the pleural fluid and was the commonest pathogen isolated from the same sample. This is similar with study from South India 23.6% (14). This result is in discrimination with other studies, where E.coli is the frequently isolated from pleural fluid (13),(18). In this study, from the peritoneal fluid frequently isolated GNB was E.coli followed by Klebsiella species. This is concordance with studies from South India (14),(17). In India, from different geographical areas E.coli was reported as predominant isolated organism from peritoneal fluid (18),(20). As E.coli is a normal flora of intestinal tract as well as facultative anaerobe so easily grow in routinely used media in laboratory. In present study, susceptibility pattern of isolates were good. GNB were highly sensitive towards carbapenem groups, susceptibility for amikacin and gentamycin is reduced. Third generation cephalosporin (ceftazidime and ceftriaxone) was showed less effective. Gram positive organisms were entirely sensitive towards vancomycin, levofloxacin, linezolide, teicoplanin and decreased sensitivity seen for aminoglycoside group (amikacin and gentamycin). Antimicrobial pattern of previous studies described in the [Table/Fig-7,8] (6),(13),(14),(16),(17),(18). In this study, all isolates of P.aeruginosa were sensitive to tigecycline and colistin, susceptibility to carbapenem antibiotics were meropenem 80% and imipenem 60%, Piperacillin-tazobactem 75% and Ticarcillin-clavulinate 70%. Susceptibility reduced towards aminoglycoside compounds; still better for amikacin than gentamycin (amikacin 75% and gentamycin 45%). Resistance was higher among cephalosporins (ceftazidime 40%, cefepime 30%) and cotrimoxazole 40%. Similar pattern of susceptibility was seen in previous studies (13),(14),(18),(17). This was different from Vijaya Durga S and Anuradha B, studies, they reported higher sensitivity to Piperacillin tazobactam 95% and >80% to carbapenems (17). In the present study, all Klebsiella pneumoniae isolates were sensitive only for reserved antibiotics, such as tigecycline and colistin, whereas susceptibility to carbapenem group was; for meropenem and ertapenem 88% both. That is comparable with study done by Sharma R and Anuradha ND (16), whereas contemplate with Madigubba H et al., study (14). Susceptibility reduced towards routinely used antibiotics such as for third generation cephalosporin (ceftazidime 53% and ceftriaxone 29%), Amoxyclave 59%, Piperacillin-tazobactam 29% and cotrimoxazole 29%. Among Aminoglycoside group susceptibility to amikacin 71% and gentamycin 53%, which was compatible with Madigubba H et al., study (14). In the present study, MRSA isolation were 4 (6.3%), all the isolates were sensitive to vancomycin, linezolide, teicoplanin and levofloxacin. Susceptibility pattern of the antibiotics were similar but isolation rate of MRSA was 27% in Mediguppa’s study, which was higher than finding in the present study (14). Limitation(s) This was a single institute based study, to make more convenient for clinicians and fabricating of effective antimicrobial policy additional data is needed. It is important to perform research work by collaboration of more than one institute. | |||||||||||||||||||||||||||||||||||||||||||
CONCLUSION | |||||||||||||||||||||||||||||||||||||||||||
Significant number of both Gram negative and Gram positive microorganism isolated among various body fluids. P.aeruginosa was the commonest pathogen isolated in the present study. AMR is higher among various isolates. Over use and misuse of antimicrobials, lack of awareness of antimicrobial use, non following of infection prevention practices are the common reasons of emergence of AMR. Hence, surveillance of bacteriological profile and antibiotic susceptibility pattern of isolates from body fluids is an essential part for the selection of the most appropriate empiric antibiotic regimen which helps the clinicians to treat effectively and thus prevent morbidity and mortality associated with these infections. | |||||||||||||||||||||||||||||||||||||||||||
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