Isolation, Identification and Antimicrobial Sensitivity of Bacterial Isolates from Pus, Sputum and Urine Samples

A total of 8 different microbial organisms (7 bacteria and 1 Candida species) were isolated from 69 positive samples out of 155 clinical samples specimens of pus, urine, sputum and other body fluid culture collected from public health Civil Hospital Vyara.The most common isolates observed in our study were Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, S.pyrogens, Proteus ,Enterococcus spp, and Candida species. Staphylococcus aureus showed highest sensitivity towards Linezolid (92.3%). Streptococcus pyrogenes was found to be 100 % sensitive towards Vancomycin, Cefuroxzime, Cefazoline,Teicoplanin and 100% resistance towards Erythromycin,Ofloxacin, Ciprofloxacin, Cefadroxil, Gentamycin, Ampicillin + Sulbactam, Cloxacillin/ Oxacillin, Levofloxacin, Cefdinir and Linezolid. Enterococcus showed 100% sensitivity towards Ampicillin + Sulbactam and Moxifloxacin and moderate towards the rest of the antibiotics. E.coli was 90 % sensitive to most of the antibiotics. Pseudomonas Spp showed 100% sensitivity towards Ceftriaxone, Gentamycin,Piperacillin + Tazobactam. Klebsiella Spp showed highest sensitivity (89.5%) towards Ciprofloxacin. Introduction Infectious diseases are a major cause of death around the world and are responsible for affecting the living conditions of millions of people. Microbes (bacteria, fungi, parasites and viruses) cause infectious diseases and antimicrobial agents (such as penicillin, streptomycin and other antimicrobial) have been developed to combat the severity and spread of many of these diseases. The use of antimicrobial agents for prevention or treatment of infections in humans in any dose and over any time period cause a reduction in microbial populations. The emergence of resistance to antimicrobial in previously susceptible bacterial pathogens is a major challenge to infectious disease. Antibiotics were considered to be the most effective therapeutic agents to combat microbial infections. But due to the overuse of antibiotics, an emergence and spread of multidrug resistant strains among different groups of microorganisms have taken place. Infections resistant bacteria are emerging threat all over the world both as hospital acquired as well as community acquired microorganisms The antimicrobial resistance is one of the main problems in clinical as well as public health view points. The antimicrobial resistance is not only increasing morbidity and mortality but also great economic loss encompassing use of more expensive antibiotics to treat infection as well as threat of resistance to them. [1] When immunity is decreased or humans are attacked by virulent bacteria resulting in respiratory tract infection, urinary tract infection and gastrointestinal infection. Antimicrobials like third generation cephalosporines are used to cure these infections. These drugs are highly active against gram-negative cocci, gram-negative bacilli and anaerobes. Hence, there is a need to conduct area specific infections and their resistance patterns, so as to generate data that would help clinicians to choose the correct therapy. Antimicrobial resistance in E. coli has been reported worldwide and increasing rates of resistance among E. coli is a growing concern in both developed and developing countries. A rise in bacterial resistance to antibiotics complicates treatment of infections. [2] Recently, WHO warned the community that Multi-drug resistant bacteria are emerging worldwide, which is a big challenge to healthcare and if immediate action is not taken, then antibiotics may lose their power to cure diseases. In India, the reasons for development of antimicrobial resistance could be due to irrational use of antibiotics, over the counter availability of higher or broader antimicrobial agents, higher prevalence of infection and poor monitoring of antibiotic susceptibility in hospitals. [3] Drug resistance among gram negative bacilli is of clinically importance and pose serious threat to public health. Numerous studies have been performed to identify susceptibility patterns of gram negative bacterial isolates. The present study was done to isolate,identify and determine the sensitivity of the isolates fro different clinical samples of Civil Hospital,Vyara. MATERIALS and METHODS Sample collection A total number of 155 Pus, Sputum,Urine and other body fluid samples were included in the study.The above samples were taken from patients who came for treatment to Civil Hospital Vyara. All clinical samples were collected and processed according to standard operating procedures. Isolation and Identification of pathogenic isolates Clinical isolates were identified by their morphological characteristics on MacConkey agar, Blood agar medium and Nutrient agar. The colonies obtained on the respective media were processed for Gram Staining. The biochemical tests performed were Catalase test, Coagulase test, Indole test, Oxidase test, Citrate Test and Urease test Volume : 6 | Issue : 6 | June 2016 | ISSN 2249-555X | IF : 3.919 | IC Value : 74.50 ReseaRch PaPeR 762 X INDIAN JOURNAL OF APPLIED RESEARCH Antibiotic susceptibility tests The identified isolates were then subjected to sensitivity test on Muller Hinton Agar The using the Kirby-Bauer disk diffusion method. [4] The commercial available antibiotic discs used for the study were Amikacin, Amoxycillin, Ampicillin, Azithromycin,Amoxicillin + Clav, Ampicillin + Sulbactam, bacitracin, Ciprofloxacin, Cefixime,Cefadoxyl, Cefotoxamine, Cefalaxine, Cetriaxone, Cefepime, Cefazoline, Cefuroxzime, Cefdinir, Cefalaxine, Ceftazidime, Cloxacillin / Oxacillin, Erythromycin, Gentamycin, Imipenem, Levofloxin, Linezoid, Moxifloxacin, Norfloxacin, Nitrofurantoin, Optochin, Ofloxacin, Penicillin G, Piperacillin+Tazobactam,Trim ethoprim+Sulfa,Teicoplanin,Tetracycline,Vancomycin. Result and Discussion Collection of sample, isolation and identification of pathogenic bacteria and yeasts from different clinical samples and specimens:Out of the 155 specimens, 69 (44.51%) specimens were found to have microbial pathogens. A total of 8 different microbial organisms (7 bacteria and 1Candida species) were isolated in 69 specimens. (Table No:1 and Table No:2) Table No1: Number and Frequency of positive and negative cases for microbial growth from different clinical samples and specimens Sr.No Source of samples and specimens Total No of positive Cases % No of Negative Cases 1 Body Fluid Urine Samples 20 9 13.04 11 2 Pus (wound) swarb 40 31 44.93 9 3 Respiratory tract: Sputum 48 22 31.88 26 4 Other body fluid 47 7 10.14 40