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DETECTION & PREVALENCE OF EXTENDED SPECTRUM βETA - LACTAMASES AMONG ENTEROBACTERIACEAE SPECIES FROM VARIOUS CLINICAL SAMPLES AT KIMS, AMALAPURAM. *N.Padmaja,Anand Acharya, P.N.Rao. Abstract: The present study was conducted in the Department of Microbiology, KIMS, Amalapuram, East Godavari from January 2012 to July 2012. Out of 100 different clinical samples, 50 were culture positive. Of the 100 samples collected, more were from post operative wound sepsis - 44 (44%), followed by cellulites - 20 (20%), Ulcers - 17 (17%), Injuries 15 (15%). Least number of cases are from burns - 4 (4%). Among 50 culture positive cases, 38 (76%) isolates belonged to Enterobacteriaceae family, followed by Pseudomonas aeruginosa - 8 (16%), followed by Staphylococcus aureus - 4 (8%). Among 38 Enterobactriaceae family isolates. 15 were ESBL producers. Among ESBL positive strains, more drug resistance was seen to ceftazidime and Ampicillin (93.33%), followed by Ceftrioxone (86.66%), Aztreonam & Cefotaxime (80%). All ESBL producers showed full sensitivity to Imipenem. Introduction: Hospital outbreaks of multi-drug resistant Enterobacteriaceae are now frequently caused by extended spectrum beta-lactamase (ESBL) producers. The incidence of ESBL producing strains among clinical isolates has been steadily increasing over past years resulting in limitation of therapeutic options. β - lactamase antibiotics are the most varied and widely used agents accounting for over 50% of all systemic antibiotics in use. The most common mode of acquisition of bacterial resistance to β - lactam antibiotics is the production of β – lactamase. However, new β - lactamases emerged against each of the new classes of β – lactamase that were introduced & caused resistance. The latest in the arsenal of these enzymes has been evolution of extended spectrum β - lactamases (ESBLs). These enzymes are commonly produced by many members of Enterobacteriaceae and efficiently hydrolyze oxyimino cephalosporins conferring resistance to 3rd generation cephalosporins like Cefotaxime, Ceftazidime & Ceftriaxone. ESBLs pose a major problem for clinicians. Hence, this study is taken to identify the prevalence of these strains in hospital & to determine suitable preventive measures & treatment policies. Materials & Methods: Different clinical samples collected from both inpatients & outpatients of KIMS Hospital, Amalapuram, during January 2012 - July 2012, constitute the material for this study. A total of 38 Enterobacteriaceae species were isolated from 100 pus samples of different diseases including both sexes and all age groups. Materials Required: Specimens were inoculated on to the media – Mac Conkey, Blood agar and incubated at 37 C for 18 - 24 hr and identified by Gram’s stain, culture, biochemical tests and Antibiotic sensitivity test. For detection of ESBL, ESBL E Strip test is used. Method : E-TEST ESBL SCREEN : (CORMICAN etal 1996) 0 The strip carries two antibiotic gradients Ceftazidime (0.5-35 µg/ml) and Ceftazidime with Clavulanic acid (0.064 - 4 µg/ml). The E test method was carried out according to manufactures instructions. Colonies from 18-24 hrs. culture were emulsified in sterile normal saline, incubated for 4 to 6 hrs. to an inoculum turbidity equivalent to 0.5 Mc Farland standard. This suspension was swabbed over on to Muller hinton agar plate and it is allowed to dry for 10 – 15 mins at room temperature. An ESBL E strip was then applied to plate with sterile forceps and the plate was incubated at 370 C for 18 hrs. After incubation, the MICs were read according to the manufacturers guidelines and the ratio determined. (The MIC ratio ≥ 8 indicates ESBL production. It is considered Gold standard.) INTERPRETATION OF TEST RESULT: The MIC value was read where the elliptical zone of growth inhibition intersected the E test strip. The MIC of Ceftazidime alone on one end of strip was compared with the MIC of Cetazidime + Clavulanic acid combination at other end. A greater than four fold reduction in the Cetazidime MIC in the presence of Clavulanic acid was taken as positive for ESBL production. For ESBL screening, the antibiotic discs used are Ceftazidime, Cefotaxime, Ceftrioxone, Aztreonam, Amoxycillin+Clavulanic acid, Ceftazidime+Clavulanic acid, Cefotaxime+Clavulanic acid and Imipenem. The incubated plates were read for zone of inhibition and results are interpreted comparing with the standard zone size given by Kirby-Bauer chart Quality control: Escherichia coli ATCC 25922 - Used as negative control. In house ESBL producers are used as positive control. Results: Out of 100 samples collected, high number of cases are from males 70 (70%) and low number of cases are from males 30 (30%). S.No Sex No. Of Cases % 1 Males 70 70 % 2 Females 30 30 % Among 100 sample collected, more were from post-operative wound sepsis - 44(44%), followed by Cellulitis - 20 (20%), Ulcer - 17 (17%), Injuries 15 (15%), Burns 4 (4%). S.No Disease No. of Samples 1 Post operative wounds 44 2 Cellulitis 20 3 Ulcer 17 4 Injuries 15 5 Burns 4 Of the 100 samples processed, 50 were culture positive. Of these, 38 (76%), isolates belonged to Enterobacteriaceae family, followed by Pseudomonas aeruginosa 8 (16%), followed by Staphylococcus aureus - 4 (8%). Table: 3 Different types of Bacterial species isolated S.No. Bacterial Species Culture Positive cases Percentage 1 Escherichia coli 21 42 % 2 Klebsiella pneumonia 9 18 % 3 Proteus mirabilis 4 8% 4 Citrobacter 2 4% 5 Proteus vulgaris 2 4% 6 Pseudomonas aeruginosa 8 16 % 7 Stahylococcus aureus 4 8% Of the 38 Enterobacteriaceae isolates, 15 are ESBL producers. Table 4: Distribution of Bacteria producing ESBLs S.No Bacterial Species Total isolates ESBL Producers No. % 1 Escherichia coli 21 8 38 % 2 Klebsiella pneumoniae 9 5 55.5 % 3 Proteus mirabilis 4 1 25 % 4 Citrobacter 2 1 5% 5 Proteus vulgaris 2 0 - (Note: Percentage of ESBL producers calculated as per the respective total isolates) Among ESBL positive strains, more drug resistance was seen to Ceftazidime and Ampicillin (93.33%), followed Ceftriaxone (86.66%) Aztreonam & Cefotaxime (80 %). All ESBL producers showed full sensitivity to Imipenem. Table 5: Comparison of Antimicrobial Resistance pattern of ESBL producers & Non ESBL producers. S.No Drugs ESBL Positive (15) No. % ESBL Negative (23) No. % 1 Ampicillin 14 93.33 % 10 43.47 % 2 Gentamicin 3 20 % 9 39.13 % 3 Amikacin 4 26.66 % 4 17.39 % 4 Ciprofloxacin 4 26.66 % 3 13.04 % 5 Chloramphenicol 5 33.33 % 4 17.39 % 6 Co - trimoxazole 7 46.66 % 12 52.17 % 7 Aztreonam 12 80 % 8 34.78 % 8 Cefotaxime 12 80 % 2 8.69 % 9 Ceftazidime 14 93.33 % 3 13.04 % 10 Ceftriaxone 13 86.66 % 3 13.04 % 11 Imipenem 0 0 0 0 (Note: Percentage of positive and negative ESBL drug resistance pattern calculated as per respective antibiotics tested). Discussion: The present study was undertaken to evaluate the pattern of drug resistance produced by isolates of Enterobacteriaceae family causing various diseases especially in hospitalized patients and to known the prevalence of ESBL production. In the present study, out of 100 pus samples, rate of recovery of isolates belonging to Enterobacteriaceae family is 38. i.e. 76 %. In this study, males (70%) are more commonly affected than females (30%) due to the fact that males are more involved in out door activities & hence chances of infection or injury are more likely. An outbreak of nosocomial infection due to ESBL isolates of Enterobacteriaceae family occur world wide in intensive care units as shown by different workers, Yagi et al 2000, Silver et al 2001. Risk factors associated with ESBL production include – prolonged hospital or intensive care stay, use of multiple courses of antimicrobial therapy, particularly extended spectrum cephalosporins. In the present study, ESBL production is more in Cellulitis cases due to prolonged hospital stay & indiscriminate use of multiple courses of antibiotics. Hence indiscriminate use of cephalosporins and broad spectrum antibiotics should be avoided. References: 1. A. Varaiya, J. Dogra, M. Kulkarni, P. Bhalekar - " Extended spectrum Beta lactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae in Diabetic foot infection Department of Microbiology ”, S.L. Raheja Hospital, Mahim West, Mumbai - 400016, IJMM, India - 7th August 2008. 2. Subha, S. Ananthan – “ Extended spectrum Betalactamase (ESBL) Mediated Resistance to Third Generation Cephalosporins among Klebsiella pneumoniae in Chennai “ - Indian Journal of Medical Microbiology, (2002) 20 (2) ; 92-95. 3. A.A. Dashit, P. West, R. Paton and S.G.B. Amyes – Characterization of extendedspectrum Beta-Lactamase (ESBL) - Producing Kuwait and UK strains identified by the Vitek system, and sub sequent comparison of the Vitek system, with other commercial ESBL - Testing systems using these strains” – Molecular Chemotherapy, Centre for Infectious Disease – January 2006. 4. Amit Jain & Rajesh Mondal – "Prevalence and antimicrobial resistance of extended spectrum Beta-Lactamase producing Klebsiella spp. isolated from cases of neonatal septicemia" – Indian J Med. Res, January 2007, PP 89-94. 5. Ankur Goyal, Amit Prasd, Ujjala Ghosal & K.N. Prasad - "Comparison of disk diffusion, disk potentation & double disk synergy methods for detection of extended spectrum beta lactamase in Enterobacteriaceae" – Indian Journal Med Res. 128, August 2008, PP 209-211. 6. Ashwin N Ananthakrishnan, Reba Kanungo, A. Kumar and S. Badrinath - "Detection of extended spectrum Beta-lactamase producers among surgical, wound infections and Burns patients in jipmer" - Indian Journal of Medical Microbiology (2000) 18 (4) : 160165. 7. Bisson, G.Fishman, N.O. Patel, J.B.; Edel Stein, P.H ; Lautenbanch,E. '' Extendedspectrum beta-lactamase-producing Escherichia coli and Klebseilla species; risk factors for colonization and impact of antimicrobial formulary interventions on colonization", Prevalence, Infection and control, Thorofare, v.23, p.254-260,2002. 8. C. Rodrigues, P Joshi, SH Jain, M Alphonse, R. Radhakrishnan, ''A Metal-detection of Beta-lactamases in Nosocomial Gram Negative Clinical Isolates''- Indian Journal of Medical Microbiology, (2004) 22 (4) : 247-250. 9. Colonder, R.; Raz, R.; Chazan, B.; akran, W. ''Susceptibility pattern of extended-spectrum Beta-lactamase producing bacteria isolated from inpatients to five antimicrobial drugs in community hospital in Northern Israel'', International Journal of Antimicrobial Agents, Shannon, v. 24, p. 409-410,2004. 10. Cormican MG, Marshall SA, Jones RN. ''Detection of spectrum β-lactamases (ESBL) producing strains by E-test ESBL screen'' J. Clin Microbiol 1996: 1880-84. 11. MS Kumar, V. Lakshmi, R. Rajagopalam ''Occurance of extended spectrum BetaLactamases among Enterobacteriaceae SPP isolated at a Tertiary care institute'' - Indian Journal of Medical Microbiology (2006) 24 (3) : 208-11. 12. Yagi, T.; Kurokawa, H.; Shibata, N.; Shibayama, K.; Arakawa, Y. ''A Preliminary survey of extended-spectrum Beta-lactamases (ESBLs) in clinical isolates of Klebsiella pneumoniae and Escherichia coli''. FEMS Microbiology Letters, Amsterdan, v.184, p.53,2000. 13. Silva, J.; Gatica, R.; Aguilar, C.; Becerra, Z.; Ramos, U.G.; Velazquez, M.; Miranda, G.; Leanos, B.; Solorzano, F.. Echaniz, G. ''Outbreak of infection with extended-spectrum Beta-lactamase producing Klebsiella pneumoniae in a Mexican hospital''. Journal of Clinical Microbiology, Washington, V.39, p. 3193-3193, 2001. A study on Bacterial contamination of Cell phones of Health Care Workers (HCW) at KIMS Hospital, Amalapuram. *N.Padmaja, P N. Rao. Abstract: A random 100 samples of cell phones of HCWs workers and Doctors working in various wards, Opds and laboratories, Blood Bank, Causality, ICU of KIMS Hospital were subjected to bacterial analysis by conventional methods to know about the Bacterial contamination of the cell phones from July 2011 to December 2011. The commonest organism isolated from the contaminated cell phone is MRSA - 20%, followed by MSSA - 5%, CONS - 10 %, Micrococcus - 15 % . The obvious observation is none of the doctors’ cell phones are contaminated. In the present study, the contamination rate is compared to the controlled group consisting patients, relatives attending in OPD’s and not working in health care setting. Introduction: Hospital infections pose problem of increased mortality and morbidity in patients. The hands of the health care workers act as an important role in transmission of hospital infections. For more than one decade cell phones have become an accessory in day to day life and they have become a part of life. The cell phones of health care workers act as a source of hospital infections. In this connection, it is imperative to study about the contamination of the cell phones which act as a vehicle of transmission of hospital infections. The causative agents of hospital infections have found a significant way to spread infection in the hospital environment. The present study is aimed at investigating the rate of bacterial contamination of the cell phones of the health care workers in KIMS, Amalapuram. In the present study the contamination rate is compared to the controlled group consisting patients, relatives attending in OPD’s and not working in health care setting. Materials and Methods: Random sampling of 100 cell phones from HCWs working in KIMS hospital from July 2011 to December 2011 was carried out. Samples were collected from OPD’s, in patients wards, ICU, Burns ward and Laboratories. A sterile wet swab moistened with sterile demineralised water is used for collecting the sample from the sides of the cell phone and key pad of the cell phone. The swabs were inoculated and streaked on 5 % Sheep blood agar and MacConkey agar (Hi Media, India). The culture plates were aerobically incubated at 370 C for 24 hrs. Isolated organisms were processed according to the colony morphology and Grams stain. The isolates were identified according to the Standard protocol. Tests for identification of Gram positive cocci included catalase, Oxidative/ Fermentative test, aerobic mannitol fermentation and coagulase production, Oxacillin sensitivity of Staphylococcus aureus was carried out by using Oxacillin disk diffusion test. Out of 100 HCWs, Doctors - 25, Nurses - 25, Laboratory Technicians - 25, House Keeping - 25. Area wise distribution of sample is as follows OPD’s : Pediatrics-2, Obg & Gyn-3, Medicine-2, Dermatology-2, Pulmonary medicine-2, Surgery3, ENT-2, Dental-2, Orthopedics-3, Ophthalmology-1, Causality-2, ICU-1. Wards : Pediatrics-2, Labor room-2, Medicine-3, Gynaecology-4, Blood bank-2, Surgery-2, ENT-2, Dental-2, Orthopedics-2, Ophthalmology-1, Burns ward-2, ICU-1. Laboratories: 25 from laboratories (Microbiology, Pathology, Biochemistry, Radiology, Blood Bank) Biochemistry-5, Microbiology-6, Pathology-9, Radiology-2, Blood Bank-3. House Keeping - 25 In the present study, the contamination rate is compared to the controlled group consisting patients, relatives attending in OPD’s and not working in health care setting. Results: Bacteriological analysis: Out of 100 samples 50 (50%) were contaminated with bacteria. Out of 50 isolates from cell phones the following bacteria were isolated Methicillin resistant Staphylococci (MRSA) - 20%. Methicillin Sensitive Staphylococci (MSSA) - 5%. Coagulase Negative staphylococci (CONS) 10% Micrococcus - 15 %. From the above observations, it is clear that 25 % of the cell phones are contaminated with hospital associated infections that is Staphylococcus aureus. . The pathogen staphylococcus aureus isolated from cell phones of Doctors is nil. Nurses18, Laboratory workers-7, House keeping-10. Area wise distribution of MRSA shows: 8 from HCW working in OPDs, 6 from Laboratories, 5 from wards, 1 from causality. Controls of 50 samples were taken. Out of 50 control samples, 1 swab from control group showed isolation of Coagulase negative staphylococci. Remaining were sterile. From the above observations it is obvious that Gram negative bacteria and Enterococcus bacteria were not isolated in any of the cell phones and it is found that the hands of HCWs at KIMS hospital setting and the cell phones used by HCWs are colonized with Staphylococci. Khivsara et al. reported 40% contamination of mobile phones by Staphylococcus and MRSA from HCWs working in a Mangalore Hospital. In a similar study from Turkey hospital, only 9% of mobile phones sampled showed contamination by bacteria associated with nosocomial infections. Similarly, Brady et al. said 14 % of mobile phones showed growth of bacteria known to cause nosocomial infection. Comparing these studies with our study, a higher percentage (50%) of Cell phones sampled was contaminated and 20% HCWs had MRSA growing on their Cell phones. DISCUSSION The above study reveals cell phones as potential threat in infection control practices and causes health care associated infections. Cell phones carried these bacteria because count of these bacteria increase in high temperatures and our cell phones are ideal breeding sites for these microbes as they are kept in warm in our pockets and hand bags. There are no guidelines for the care, cleaning and restriction of cell phones in our health care system. So in a country like India, cell phones of HCWs play an important role in transmission of infections to patients which can increase the burden of health care. CONCLUSION In conclusion, it can be said that hand hygiene is overlooked and is under emphasized in health care settings. Simple measures like increasing hand hygiene and regular decontamination of cell phones with alcohols, disinfectant wipes may reduce the risk of cross contamination by these devices. References 1. Brady RR, Fraser SF, Dunlop MG, Paterson - Brown S, Gibb AP. Bacterial contamination of mobile communication devices in the operative environment. J Hosp Infect 2007; 66:397-8. 2. Clinical Laboratory Standards Institute, Performance standard for antimicrobial disk susceptibility tests; Approved standards, 2005, vol.25, 8th edn, M02-A8. 3. Brady RR, Wasson A, Stirling I, McAllister C, Damani NN. Is your phone bugged? The incidence of bacteria known to cause nosocomial infection on healthcare worker’s mobile phones. J Hosp Infect 2006;62:123-5 4. Karabay O, Kocoglu E, Tahtaci M. The role of mobile phones in the spread of bacteria associated with nosocomial infections. J infect Developing countries 2007; 1:72-3. 5. Khivsara A, Sushma T, Dhanashree B. Typing of Staphylococcus aureus from mobile phones and clinical samples. Curr Sci 2006;90:910-2. 6. Gupta V, Datta P, Singla N. Skin and soft tissue infection : Frequency of aerobic bacterial isolates and their antimicrobial susceptibility pattern. J Assoc Physicians Indians India 2008;56:389-90. 7. Jeske HC, Tiefenthaler W, Hohlrieder M, Hinterberger G, Benzwer A. Bacterial contamination of anesthetist’s hands by personal mobile phone use in the operation theater. Anesthesia 2007;62:904-6. IDENTIFICATION OF UROVIRULENT MARKERS IN UROPATHOGENIC E.COLI * N. Padmaja, Anandacharya, P.N.Rao. Abstract: The present study was conducted in the Department of Microbiology, KIMS, Amalapuram, East Godavari District from August 2011 to January 2012. 50 E.coli strains isolated from urine samples of different clinical entities and 25 feacal isolates were studied for the detection of virulence markers of E.coli. There are 27 UPEC isolates from 50 E.coli & 5 UPEC from 25 controls. Among isolates tested the most common virulent marker is haemolysin 21 (42%), followed by Mannose resistant haemagglutination 16 (32%), cell surface hydrophobicity 13 (26%). In this, there are 14 cases with only one virulence marker, 8 with 2 marker combinations and 15 cases with combination of 3 markers. Introduction: UTI is one of the most important causes of morbidity and mortality. E.coli is the most frequent urinary pathogen isolated from 50 % - 90 % of all uncomplicated urinary tract infections. It is now recognized that there are subsets of faecal E.coli, which can colonize periurethral area, enter urinary tract and cause symptomatic diseases. These are currently defined as Uropathogenic E.coli. It has been traditionally described that certain serotypes of E.coli were consistently associated with uropathogenicity and were designated as uropathogenic E.coli. These isolates express chromosomally encoded virulence markers. The virulence factors include different adhesins, haemolysin production, haemagglutination, cell surface hydrophobicity. The present study was designated to determine the urovirulence factors of E.coli isolated from the patients of UTI and to study their antimicrobial susceptibility pattern. Materials and Methods: The study was conducted in the Department of Microbiology, KIMS, Amalapuram, East Godawari from August 2011 to January 2012. 50 E.coli strains isolated from urine samples & 25 faecal isolates were studied for 1. α Haemolysin on 5% sheep blood agar. 2. Mannose Resistant Haemagglutination. 3. Cell surface hydrophobicity. 4. Antibiotic susceptibility testing by Kirby Bauyer disc diffusion method. Inclusion criteria: Adult patients with UTI attending various clinical departments of KIMS and adult healthy individuals for stool samples are taken. Samples: 1. Clean catch midstream urine samples from the patients (50). 2. Stool samples of adult healthy individuals (25) Collection and Culture of sample: Sample is transported to Microbiology lab with in half an hour and processed by 1. Wet film preparation. 2.Culturing on Mac Conkey, Blood agar & CLED agar medium & incubated aerobically at 370 C for 24 hrs. 3. Growth on plates and significant bacterial count. i.e. 105 col / ml and tested for further identification of E.coli by various biochemical reactions. Such E.coli were screened for virulence markers. Haemolysin: For detecting haemolysin, 5% sheep B.A is used and a zone of lysis around each colony is observed after overnight incubation at 370C, if haemolysin is produced. Haemagglutination: HA is detected by clumping of erythrocytes by bacterial fimbriae in the presence of D-Mannose. The test was carried out as per the direct bacterial HA test slide method and mannose sensitive and resistant HA tests. HA was considered when it occurred in the presence of 2 % D- Mannose and Mannose sensitive, when it was inhibited by D-Mannose. Cell surface hydrophobicity: This was done by Salt Aggregation test. Strains are considered hydrophobic, if they are aggregated in concentration of < 1.4 M. Ammonium sulphate. Antibiotic susceptibility testing: This was performed on all isolates of E.coli by Kirby-bauyer`s disc diffusion method on Muller hinton agar. Results: Of the 50 patients, 0 - 15 age group comprised of 11 persons, 16 - 40 age group comprised of 18 persons and > 40 years group comprised of 21 persons. Females (30) are more than male (20) patients. Among various clinical entities, 30 cases presented with lower UTI (60%), 14 cases with asymptomatic bacteriuria (28%) and 6 with pyelonephrities (12%). Among 50 cases tested 27 (54%) were positive for virulence markers and out of 25 controls, 5 (20%) were positive for virulence markers. Among the isolates tested, the most common virulent marker is haemolysin 21 (42%) followed by Mannose resistant Haemagglutination 16 (32%) and Cell surface hydrophobicity 13 (26%). In control group, the occurrence of haemolysin was 1 (20%), MRHA 3 (60%), CSH 1 (20%). (Note: Percentage calculated as per total 50 isolates). There are 14 cases positive for 1 marker, 8 cases positive for 2 markers and 5 cases positive for 3 markers. Table showing No. of positive virulence markers - Table I Composition No. of Tested Positive virulence markers % Cases 50 27 54% Control 25 5 20% Antibiotic Sensitivity Test: Table II S.No. 1 Drug Amikacin Sensitivity % Resistant % 41 82% 6 12% 2 Nitrofurantoin 39 78% 11 22% 3 Cefotaxime 36 72% 14 28% 4 Cephalexin 26 52% 21 48% 5 Co-trimoxazole 23 46% 17 54% 6 Cefuroxime 16 16% 34 68% 7 Ampicillin 13 26% 37 74% 8 Norfloxacin 10 20% 40 80% 41% isolates were sensitive to Amikacin, 78% to Nitrofurantion, 72% sensitive to Cefotaxime, 52% sensitive to Cephalexin, 46% sensitive to Co-trimoxazole. High resistance is seen for Norfloxacin 80% followed by Ampicillin 73%, Cefuroxime 68%. (Note : Percentage drug sensitivity and resistance pattern calculated for total number of 50 strains isolated). Discussion: The present study was undertaken to study the virulence factors namely haemolysin production, MRHA and CSH. 50 E.coli isolates from urine of cases with clinically diagnosed UTI are taken as study group and 25 E.coli from faeces of healthy persons as control group. Most of the cases of UTI are from above 40 years age group (21), followed by 16-40 years(18) and 0-15 years (11). The high incidence of UTI was observed above 40 years age group followed by 16-40 years age group. This is because, the elderly patients are likely to be predisposed to conditions like 1. Urinary tract obstruction. 2.Poor bladder emptying 3. Diabetes 4. Prostate enlargement. These factors favour colonization of bacteria and play an important role in UTI. High incidence of UTI is observed in females (30) than males (20), which is due to 1. short urethra. 2. close proximity of urethra to perianal region. Regarding AST, most of the isolates are sensitive to Amikacin (82%) and Nitrofurantoin (78%). Where as most of the isolates are resistant to Norflox, Ampicillin, Cefuroxime and Cephalexin. This shows that Aminoglycosides and Nitrofurantoin are useful for most of UTI and high level of resistance indicates the lack of rational drug use. References: 1. Orskov, F.1984, Genus I.Escherichia castellani and Chalmers 1919. In: Krieg, N.and Holt, J.G. (eds), Bergey`s Manual of systemic bacteriology. London: Williams & Wilkins, 420-3. 2. Smith HW. The haemolysins of Eschericha coli. J. Pathol Bacteriology 1963; 85: 197-211. 3. Kaijser B.Immunology of Escherichia coli. K. Antigen and its relations to UTI. J. Infectious diseases 1973;127:672-677. 4. Quackenbush RL, Falkow S. Relationship between colicin V activity and virulence in E.coli. Infect Immune 1979;24:562-564. 5. Hageberg L, Jodal U, Korhonen Tk, Lidin – Janson G, Lindberg U, Svanborg Eden C. Adhension, haemagglutination and virulence of E.coli causing Urinary track infections. Infect Immune 1981;31:564-570. 6. Lindahl M, Faris A, Wadstrom T, Hjerten S. A new test based on salting out to measure relative surface hydrophobicity of bacterial cells. Biochim Biophys Acta 1981;677:471-6 7. Orskov I, Orskov F; Birch-Andersen A, Kananmori M, Svanborg-Eden C, O, K, H and fimbrial antigens in E.coli serotypes associated with pyelonephritis and cystitis. Scand J Infectious diseases 1982; Suppl 33:18-25. 8. Cavalieri SJ, Bohach GA, Snyder IS. E.coli alpha haemolysin: Characteristics and probable role in pathogenicity. Microbiol Rev 1984;48:326-343. 9. Holmes, B and Gross, R.J. 1990. Coliform bacteria: various other members of the Enterobacteriaceae. In: Parker, M.T. and Collier, L.H. (eds), Topley & Willson`s Principles of bacteriology, Virology and Immunity, 8th edn. London: Edward Arnold, 415-41. 10. L.Siegfried, Marta kmetova, Hana puzova, Maria Molokacova and J.Filika. Virulenceassociated factors in E.coli strains isolated from children with urinary tract infections. Journal of Medical Microbiology – Vol.41(1994); 127-132. 11. Johnson J: Microbial virulence determinats and pathogenesis of Urinary tract infection. Infect Dis Clin North Am 2003, 17:261-278.