MY -07 RHINOENTOMOPHTHOROMYCOSIS DUE TO CONIDIOBOLUS CORONATUS.

Anil Kumar,Vineeth Vishwam, Sukhmani Regi, Aswathy N, Kavitha Dinesh, Shamsul Karim, Department of Microbiology, Pathology & ENT, Amrita Institute of Medical Sciences, Ponekkara, Kochi, Kerala, India-682041. Background: Entomophthoromycosis is a rare subcutaneous infection manifesting either as a painless nodule on the trunk or as rhinofacial zygomycosis involving the nasal cavity, paranasal sinuses and the soft tissue of the face. The disease is largely restricted to the tropical regions of Africa, Asia and South America. Generally, infections due to Zygomycete fungi of the Mucorales order (ie, Rhizopus, Mucor, Absidia, Cunninghamella, etc) occur in immunocompromised individuals or patients with poorly controlled diabetes mellitus. These characteristically are angioinvasive, frequently disseminate, and have a rapid and often fatal clinical course. Conversely, diseases caused by Zygomycetes of the Entomophthorales order (Conidiobolus and Basidiobolus) most commonly present in immunocompetent hosts, predominantly involve subcutaneous tissue, have a protracted and chronic clinical evolution and respond very well to treatment with itraconazole and potassium iodide. Case report: We present a case of a 43-year-old male who was referred to us with the diagnosis of rhinofacial zygomycosis to start treatment with amphotericin B. Functional endoscopic sinus surgery revealed a polypoidal growth in the right middle meatus and maxillary antrum. Multiple histological sections of the tissue revealed eosinophilic granuloma with broad aseptate hyphae. Cultures grew a Zygomycete which was identified as Conidiobolus coronatus. Amphotericin B was discontinued and the patient was put on itraconazole and potassium iodide. The patient was completely cured after four months of therapy .A repeat endoscopy revealed clear sinuses. Conclusion: Identification of the fungal species helped us to successfully treat the patient with potassium iodide and avoid Amphotericin B, which is toxic and expensive. MY-08 PHOSPHOLIPASE ACTIVITY OF CANDIDA ALBICANS ISOLATED FROM VAGINA AND URINE SAMPLES Mrs.M.Sangeetha and Ms.N.Madhumitha, Department of microbiology, A.V.C. College (Autonomous), Mannampandal, Mayiladuthurai. Candida albicans is the most virulent among the Candida species, and can cause several forms of candidiasis in human. Several factors were described as virulent for pathogenesis of C. albicans. Some of them include the secreted aspartyl proteinases, germ tube formation, adherence to host tissues and phenotypic switching. Extracellular phospholipases in C. albicans is discussed as one of the virulence factors. Four types of phospholipases have been reported in C. albicans including phospholipase A, B, C, and D. The extracellular phospholipases of C. albicans have a significant role in the pathogenesis of infections and invasion to mucosal epithelia. And so the clinical isolates of C. albicans have higher levels of extracellular phospholipase activity. The main objective of the study is focused on extracellular phospholipase activities in different isolates of C. albicans isolated from vagina and urine samples. In addition, phospholipase activities were compared in C. albicans isolated from two different sources. In this study, phospholipase activity of isolates of C.albicans from urine and vaginitis origin was demonstrated using Sabouraud's dextrose agar supplemented with egg yolk. The phospholipase activity was detected in all tested isolates with a high level in Pz <0.70. In this study phospholipase activity with higher Pz (Zone of phospholipase activity) values was more common in vaginal isolates compared with urine isolates. Through this study, 100% clinical isolates of C. albicans from vaginitis and urine samples demonstrated phospholipase activity.

MY-09 PRELIMINARY IDENTIFICATION OF CANDIDA SPECIES FROM CLINICAL SAMPLES Mrs. J.Myla M.sc.,M.Phil(Ph.d), Miss.R.Aishwarya, Department of Microbiology, A.V.C. College (Autonomous), Mannampandal, Mayiladuthurai. Candida species are important etiological agents for hospital acquired infections especially in immunocompromised patients. Presumptive identification of non-albicans species of candida from various clinical samples was done in this study. Identify the different species of candida other than C.albicans by simple biochemical tests.Identification of different species of Candida is useful to analyse the prevalence of infection by the various species in clinical settings.Among the fungi,Candida species are the common causative agents for various hospital acquired infection,in immunocompromised patients especially patients with AIDS, Diabetes mellitus and following transplantation etc.,prolonged antibiotic therapy is also a predisposing factor for candidiasis. Candida is seen on skin and muscosal surfaces as commensal and it can cause endogenous infections. The sample were plated on Sabourauds Dextrose Agar(SDA) by streak plate technique and incubated for 72 hours.The cream coloured colonies were noted.Later,primary and other tests were performed to identify the Candida sp. In this study Candida tropicalis was the most common isolate and followed by Candida parapsilosis,Candida gulliermondii and Candida krusei. Amphotericin-B was found to be sensitive for most of the isolates. Hospitals may have more number of drug resistant microorganisms.Mostly candida species were identified, important causative agents for various nosocomial infections. Hence, successful treatment of such infection is important in reducing morbidity and mortality in hospitalized patients.

MY- 10 COMPARATIVE STUDY ON THE INCIDENCE AND OUTCOMES OF PIGMENTED VERSUS NON PIGMENTED KERATOMYCOSIS Manivasagam, C.Vaitilingam, Sabyasachi Sengupta, Krishnan Thiruvengadakrishnan, Department of Microbiology, Aravind Eye Hospital, Pondicherry, India. Aim: To determine the incidence, outcomes and establish factors determining visual prognosis of keratomycosis due to pigmented fungi in comparison with non-pigmented fungi. Methods: All culture proven cases of fungal keratitis from January 2006 to August 2008 were drawn from a computerized data base and those cases with adequate documentation were analyzed for predisposing factors, clinical characteristics, and microbiology and treatment methods. Outcomes of keratitis due to pigmented and nonpigmented fungi were compared using t-test and chi square test. Results: Of 373 cases of keratomycosis during the study period, pigmented fungi were etiologic agents in 117 eyes (31.3%) and nonpigmented fungi in 256 eyes (68.7%). Eyes with nonpigmented keratitis had significantly larger ulcers (14.96mm ) and poorer vision (1.42logMAR) at presentation compared to those with keratomycosis due to pigmented fungi (p=0.01). The characteristic macroscopic pigmentation was seen in only 14.5% in the pigmented keratitis group. Both groups responded favourably to medical therapy (78.1% v/s 69.1 %) with scar formation (p=0.32) and showed a significant improvement in mean visual acuity compared to that at presentation (p<0.01).Visual improvement in terms of line gainers and loosers in the subgroup of eyes that experienced healing was also similar. Location of the ulcer was the only factor that had significant predictive value for visual outcome (p=0.021). Conclusion: Incidence of keratomycosis due to pigmented fungi may be increasing as compared to previous data. These eyes have similar response to medical therapy and similar visual outcome compared to non pigmented keratitis. Central ulcers have a poor visual outcome.

MY-11 A SIMPLE INEXPENSIVE DIAGNOSTIC METHOD TO DIFFERENTIATE CANDIDA DUBLINIENSIS FROM CANDIDA ALBICANS FROM CLINICAL ISOLATES. Ambujavalli.B.T, Purushothaman.K, Anita.S, Anupma Jyoti Kindo m, Dept of Microbiology, Mycology section, Sri Ramachandra University, Porur, Chennai. Introduction Candida dubliniensis is a pathogenic species of Candida which shares any phenotypic features with Candida albicans including the ability to form germ tubes and chlamydospores. These similarities have caused significant problems in the identification of C.dubliniensis by the average clinical mycology laboratory. The increase in C.dubliniensis requires proper laboratory identification because it may have epidemiological and therapeutic implications.Aims and Objectives: In the present study, an attempt was made to look for Candida dubliniensis among our isolates which were identified as C. albicans. A simple inexpensive method as described by Anuradha et al was followed - to grow the germ tube positive Candida in SDA incorporated with 6.5% NaCl. Materials and Methods: The isolates of germ tube positive Candida identified as C. albicans were subcultured in SDA with 6.5% NaCl and incubated at 28 degrees overnight. Other methods to morphologically differentiate C. albicans and C. dubliniensis were also put up. Results: The germ tube positive Candida grew well in SDA with 6.5%NaCl at 28 degrees unlike the control strain of C.dubliniensis ( also germ tube positive) which did not grow in SDA with 6.5%NaCl. All our 90 isolates were identified as Candida albicans by this method. Conclusion: A simple method as described above can be used to differentiate C. albicans from C. dubliniensis in a low resource laboratory. The isolation rate of Candida dubliniensis was nil in our present pilot study.Further study with a greater sample size is necessary to know the true prevalence.

V- 12 CRIMEAN CONGO HAEMORRHAGIC FEVER IN INDIA ARE WE PREPARED AC Sharmila, V Sriandaal,E Susanblossia,B Vallab Ganesh, AUma,Tirumalaikolundusubramanian.P, Department of Microbiology, Chennai Medical College Hospital and Research Centre, Irungalur, Trichy 621 105 Back Ground: Crimean-Congo haemorrhagic fever (CCHF) virus was found to be the cause for an outbreak of a mysterious fever which killed three people at Ahmedabed in January 2011. It was reported by National Institute of Virology, Pune. The present report discusses the challenges faced by the Indian Health Care System at different levels. Transmission: Ticks are required to transmit CCHF Clinical challenges: The heamorrhagic manifestations overlap with Dengue heamorrhagic fever. Diagnostic Challenges: Diagnostic facilities required to identify the illness and its consequences are lacking in primary, secondary and tertiary care level hospitals. Therapeutics: Supportive measures are available at tertiary care centers. Antiviral drugs (Ribavarin) have to be used carefully in view of adverse effects. Risk of Infections to Healthcare workers: As Health Care Workers (HCW) are susceptible to CCHF virus through body fluids, re enforcement of Universal infection control programme has to be implemented. Reporting system: Doctors working at different levels are not aware their responsibility to report such cases to Health authorities as enforcement system is not strengthened.Remedial Measures: They have to be highlighted as Indian Health community has been facing the challenges of SARS and Bird flu.

V-13 OPPORTUNISTIC INFECTIONS IN RELATION TO ANTIRETROVIRAL STATUS AMONG AIDS PATIENTS FROM SOUTH INDIA Srirangaraj.s, Venkatesha.d, Department of microbiology, Mysore medical college and research institute, Mysore-570001. Background: There is a need to generate data from India on relative frequencies of specific opportunistic infections (OIS) in different regions and their relation to the choice of commonly used generic highly active anti-retroviral therapy (HAART) regimens. Objectives: To document the prevailing prevalence pattern of ois ,both before and after haart, to assess the risk of developing an OI after 6 months of HAART initiation and to document the risk of developing an oi in the first six months after HAART initiation for those on efavirenz (efv)- based regimens when compared with those on nevirapine (nvp) based regimens. Materials and Methods: in a prospective observational cohort study conducted in south india involving 108 haart-naive aids patients, different pathogens were isolated and identified using standard laboratory techniques. data analysis was done using spss software (version 16.0). risk of developing an oi after haart initiation was assessed using the likelihood ratio test from cox regression models. Results: Tuberculosis (53.4%), oral Candidiasis (27.2%) and Herpes zoster (14.7%) were the common infections seen. the risk of developing an oi within 6 months of haart initiation was 6.48%. time to development of an oi in the first 6 months of haart was shorter for the nvp-based regimens than with efv-based regimens, but this difference was not statistically significant (hazard ratio = 0.891, 95% ci: 0.179 to 4.429; p = 0.888). Conclusion: tuberculosis is the most important oi before initiation of HAART. both efv and nvp-based regimens are equally efficacious in controlling OIS.

V- 14 SCREENING FOR HBsAg MARKER IN CUDDALORE, TAMILNADU J. Poorani, P. Pushpavalli, T.H. Rashithatur Rahman, Department of Microbiology, St. Josephs College of Arts and Science (Autonomous), Cuddalore 607 001 The study was conducted to find out the prevalence of HBsAg marker among the population of Cuddalore. Blood samples (n=101) were collected from the people of three areas in Cuddalore and the serum was separated. The serum samples were screened for the presence of HBsAg marker by ELISA using a commercial kit (Diasorin, Italy). The study was approved by the Institutional Ethical Committee. The results were analysed with the demographic data obtained from the subjects. Out of 101 samples tested 8 (7.92 %) were found to be reactive for HBsAg. 8.86% of the tested males and 4.54% of the females were found to harbour the antigen. 29.70% (n=30) of the people tested were blood donors and 6.66% of them were tested reactive, as against 8.45% among the non-donors. The prevalence of the marker was found to be 9.43% among the unmarried where as 6.25% was found among the married. Reports by various research groups have suggested a high prevalence of Hepatitis B in India, 4.7% , according to Indian Association for Study of the Liver . The present study also correlates with this finding. The high prevalence among the study population necessitates the need for immunization against Hepatitis B virus infection.

V-15 LAY PRESS AND SWINE FLU P.Thirumalai kolundu Subramanian, M.Ismail, A.Uma, Chennai Medical College Hospital & Research Centre, Irungalur, Trichy Lay press disseminates wide variety of information to public. Newspaper reading is prevalent even in remote parts of India, despite the advances in Audio visual media. The present report discusses the matters on swine flu published in lay press, as no such studies were made from India. Objectives: 1] To analyze the pattern of published matters on Swine flu in lay press. 2] To identify the areas focused and 3] To elicit the response from the policy makers on the news published in the lay. Materials and Methods: Any information containing the word swine flu and/or H1N1 published in two widely circulated News papers [The Hindu [English] and Dinamalar [Tamil]] were considered for the study. They were searched daily in all pages from April 2009 to October 2010, and analyzed and classified by the first two and confirmed by senior author. Simple statistics was used. Results: A total of 430 matters were published in The Hindu and 207 in Dinamalar. Pictorial presentation was published more in English than Tamil news paper than Tamil. The matters were related to epidemiology [23.7/29%], diagnostic aspects [8.2/5.7%], and preventive aspects including treatment [30.9/31%], consequences [4.6/2.8%] International news [8.8/4.8%], alternative medicine [1.3/3.8%] and others [22.5/29.9%] in the Hindu and Dinamalar respectively. Comments: Lay press educated public, alerted travelers, sensitized the policymakers and health officials, provided information on preventive aspects and arranged to implement containment measures. Thus lay press was found to be a valuable way of disseminating the news to various populations living in different geographical areas of India.

AM-16 A SURVEY ON USAGE OF ANTIBIOTICS AMONG COLLEGE STUDENTS Bharathi Prabhakaran, R.S.Vigneshwari, Alwin Surya, Abinaya, Anto Venetia, Abirami, Asish, Anitha, Balaji, A.Uma, Dept of Microbiology, Chennai Medical College Hospital & Research Centre, Irungalur, Trichy-621105 Introduction: Antimicrobial resistance is a serious public health threat and concern worldwide. As inappropriate self medication and incorrect dosage/duration are considered for the emergence of antimicrobial resistance, present study was undertaken to know the status among students of higher education. Objectives: a) To identify the factors responsible for self medication among college students. b) To determine the awareness of antimicrobial resistance among them. Design and setting: The survey was carried out with a self administered, anonymous predated questionnaire to elicit their attitude and awareness on antibiotic usage. The participants included Medical, Nursing and Engineering college students of the campus of Chennai Medical College Hospital and Research Centre, Irungalur, Trichy. Results: Of the 200 Engineering, 150 medical, 44 Nursing students surveyed .the response rate among engineering, medical and nursing students were 65%, 86.6%, and 68.1 % respectively. The percentage of students who took self medication over the counter (OTC) were 70% (Nursing students), 51.5% (Engineering students) and 13% (Medical).Likewise the percentage of awareness to antimicrobial resistance among medical, nursing and engineering students was 22.3,10 and 4.6 % respectively. The difference among these groups was significant statistically. Conclusions: The teenagers and young Constitute a large section of population and since they can easily be trained and motivated for a change in them and for society, audio visual and educational program has to be designed by Ministry of Health and introduced among them through educational system for the future of India.

AM - 17
DETECTION AND CHARACTERISATION OF METALLO BETA LACTAMASES (MBL) PRODUCING PSEUDOMONAS AND ACINETOBACTER IN A TERTIARY CARE CENTRE FROM SOUTH INDIA

M.Shanthi, Uma Sekar, K.Arunagiri*, B.Sekar*, Department of Microbiology, Sri Ramachandra Medical College and Research Institute, Porur, Chennai-600116 and *Central Leprosy Teaching and Research Institute, Chengalpattu, TamilNadu. Objective: To detect the presence of MBLs by phenotypic and genotypic methods in nosocomial isolates of Pseudomonas and Acinetobacter species.Materials and methods: One hundred and seventy nine nonreplicate, consecutive, carbapenem resistant Pseudomonas aeruginosa (PA)-61, Acinetobacter baumannii (AB)-115, Acinetobacter lowfii (AL)-2 and Pseudomonas stutzeri (PS)-1 ,isolated from patients hospitalized for >48 hours were included in the study.They were obtained from blood(34), exudates (37), respiratory secretions(83) and urine(25). The Minimum Inhibitory Concentration [MIC] to Imipenem [IPM] and Meropenem [MER] were determined and interpreted according to CLSI guidelines. Double Disk Synergy Test (DDST) with Ethylene Diamine Tetra Acetic acid (EDTA) and Modified Hodge Test (MHT) were used for phenotypic screening of MBL /Carbapenamases. blaVIM and blaIMP MBLs were detected by PCR. Results: MIC to IPM and MER ranged from 8-256g/ml. DDST detected the presence of MBLs in 104(58%) isolates. PCR detected blaVIM and blaIMP in 92(51%), suggesting the presence of other MBL genes. Of the 115 AB isolates 52 had bla VIM and one isolate had both blaIMP and blaVIM genes. Of the two AL isolates, one carried the blaVIM gene. Among the PA blaVIM was detected in 34 isolates while 2 of them carried blaIMP gene. The lone PS isolate carried the bla VIM gene. MHT was positive in 158 (88%) isolates suggesting the presence of other carbapenamases. Conclusion: MBL production is an important mechanism of carbapenem resistance.Detecting their presence in clinical isolates has important implications for clinical decision making and implementing infection control measures. AM- 18 BIOFILM FORMATION AND ANTIBIOTIC RESISTANCE IN CLINICAL ISOLATES OF PSEUDOMONAS AERUGINOSA Kranthi Kosaraju*, Chiranjay Mukhopadhyay, Vandana KE, Kiran Chawla, Indira Bairy, Department of Microbiology, Kasturba Medical College, Manipal University, Manipal, INDIA. Upto 60% of all human infections are caused by biofilm producing organisms which are difficult to eradicate. Bacterial biofilms are dense communities of bacteria encased in a exopolysaccharide that adheres to biological or prosthetic surfaces. Pseudomonas aeruginosa is a known biofilm producer especially in the lung of patients suffering from Cystic fibrosis. We undertook this study to check for biofilm producing capability of Pseudomonas aeruginosa isolated from various clinical specimens and also to check their suspectibility pattern against anti-pseudomonal antibiotics. This study was conducted on 100 consecutive clinically significant isolates of Pseudomonas aeruginosa from pus, sputum, blood and urine. Isolates were identified and their antibiogram was performed following standard methods. All these isolates were tested for biofilm production by the method described by Stepanovic et al. Minimum inhibitory concentration against Ceftazidime, Ciprofloxacin, Rifampicin and Colistin was determined by Agar dilution methods. Among the isolates tested, 30% were identified as strong biofilm producers, 40% were moderate biofilm producers and the rest were detected as weak biofilm producers. 20% of the isolates were resistant to Ceftazidime, 33.3% resistant to Ciprofloxacin and 15.56% demonstrated resistance to Colistin when tested in single. Only one-third of strong biofilm producing isolates demonstrated resistance to Ceftazidime, Ciprofloxacin but remained sensitive to Colistin. Biofilm formation is commonly associated with Pseudomonas aeruginosa isolated from various clinical samples. Biofilm formation was not significantly associated with antibiotic resistance in our study, contrary to that seen in other studies.

AM- 19 DETECTION OF METALLO-BETALACTAMASES IN PSEUDOMONAS SPECIES FROM VARIOUS CLINICAL SAMPLES Syed Ali. A, Sageera Banoo, Latha. R, Rajeshwar Lall, Department of Microbioloy, Aarupadai Veedu Medical College & Hospital, Kirumampakam, Puducherry. Aims & Objectives: The emergence of carbapenem resistant Pseudomonas species due to the production of Metallo-beta-lactamases (MBLs) has become a major threat to the control of infections caused by them. Hence a prospective study was done to determine the occurrence of MBLs among Pseudomonas species in our hospital. Materials & Methods: 88 clinical isolates of Pseudomonas spp. were collected within a period of six months. MIC for Imipenem was tested using agar dilution method. Testing for MBL production was done by Imipenem EDTA double disc synergy test (DDST), Imipenem - EDTA combined disk test (CDT) & modified Hodge test (MHT). Detection of Amp C betalactamase was done by modified three dimensional tests (MTDT). Results: MIC of 12 out of 88 isolates(13.63%) showed resistance to Imipenem. 5 isolates(5.68%) which were positive by MHT were also found positive by the other two methods . 8 isolates(9.09%) were detected positive for both DDST & CDT. One isolate was found additionally positive by CDT. 3 out of 12 isolates were not found positive by any of these methods. AmpC betalactamases were also seen in 5 out of 9 MBL producing Pseudomonas. Conclusion: MBL producing Pseudomonas is prevalent in our hospital. CDT was found more sensitive & simple method when compared with the other two tests. However combinations of at least two methods are needed for the detection of MBL.

AM-20 EVALUATION OF VARIOUS PHENOTYPIC METHODS FOR THE DETECTION OF METALLO-BETA-LACTAMASE PRODUCTION IN ACINETOBACTER SPECIES. Anusheela Howlader, Sageera Banoo, Latha.R, Rajeshwar Lall, Department of Microbiology, Aarupadai Veedu Medical College &Hospital, Kirumampakam, Puducherry. Aims & Objectives: The emergence of carbapenem resistant Acinetobacter species due to the production of metallo-beta-lactamases (MBLs) has become a major threat in the control of infections caused by them. We conducted a study to evaluate the efficacy of the various phenotypic methods used for the detection of MBL production among Acinetobacter species. Materials & Methods: 22 clinical isolates of Acinetobacter were collected within a period of three months. MIC for Imepenem was tested using agar dilution method. Testing for MBL production was done by Imepenem EDTA double disc synergy test (DDST) , Imepenem - EDTA combined disk test(CDT) & modified Hodge test (MHT). Results: MIC of 8 out of 22 isolates showed resistance to Imepenem. 3 isolates which were positive by MHT were also found positive by the other two methods. 5 isolates were detected positive for both DDST & CDT. One isolate was found additionally positive by CDT. 2 out of 8 resistant isolates were not found positive by any of these methods. Conclusion: CDT was found more sensitive & simple method when compared with the other two tests. However combinations of at least two methods are needed for the detection of MBLs. Two isolates which were not detected by any of these methods indicates that genotypic assays are further needed for better detection of MBL.

AM -21 EVALUATION OF DRUG RESISTANCE IN ACINETOBACTER Abhisek Routary, Dr. Radha Madhavan, Dr. S. Gomathi,SRM Medical College Hospital & Research Centre, Department of Microbiology, Kattankulathur-603 203. Objective: To assess drug resistance in Acinetobacter isolated from different clinical samples (respiratory exudates, urine, pus & blood) and hospital inanimate environment at SRM Hospital. Method: In this study 75 isolates of Acinetobacter (55clinical and 20 environmental samples) were collected and phenotypically characterized by biochemical reaction (12 carbon assimilation test), susceptibility testing using 10 antimicrobial agent. MIC was determined for carbapenem. Result: 70% of total isolates were found to be A. baumanii, most were resistant to first line drug, 60% of total isolates found to be carbapenemase producers mainly due to metallobetalactamese production as confirmed by Imepenem EDTA method.40% of total resistant isolates showed MIC at (16 mg/L) for imiepenem, 16% of total resistant isolates showed MIC at (16 mg/L) for meropenem. Conclusion: Most of the Acinetobacter baumanii isolates were multidrug resistant in our set up.

AM -22 EXTENDED SPECTRUM LACTAMASE PRODUCING KLEBSIELLA PNEUMONIAE AND ESCHERICHIA COLI IN NEONATAL INTENSIVE CARE UNIT. Dr.N.Girish, Dr.K.Saileela, Dr.S.K.Mohanty, Dept. of Microbiology, Kamineni Institute of Medical Sciences, Narketpally, Nalgonda dist. Andhra Pradesh. Introduction: Neonatal Septicemia is an important cause of morbidity and mortality. As infections due to ESBL producing K.pneumoniae & E.coli are on the rise, the present study was carried out in the NICU of KIMS, Narketpally, with an aim to identify any environmental sources & the mode of transmission over a period of 3 years from August 2006 to July 2009. Materials & Methods: A total of 264 neonates admitted with clinical features suggestive of septicemia in the NICU were studied by blood culture and CRP estimation. Antibiotic susceptibility pattern was determined. ESBL detection was done by double disc synergy test. Environmental samples from various sites (Incubators, phototherapy units, suction apparatus, trolley, door, floor, work surfaces) were collected using sterile swabs every month and processed simultaneously. Results: Of the 264 blood cultures, 197(75%) showed bacterial growth. K.pneumoniae, 64(32.7%) was the commonest organism followed by E.coli 55(28%), S.aureus 31(16%), Pseudomonas aeruginosa 28(14%), Acinetobacter 13(7%), and Coagulase negative Staphylococci 6(2.8%) respectively. K.pneumoniae & E.coli were isolated from various environmental sites at least on one occasion and consistently from phototherapy units, door & floor of the NICU. The antibiogram of majority of ESBL producing strains of K.pneumoniae & E.coli isolated from clinical samples were similar to that of isolates from environmental sites. Conclusion: Wide spread use of third generation cephalosporins as a preemptive antibiotic for suspected cases of septicemia have contributed to the emergence of ESBL producing K.pneumoniae & E.coli in addition to other risk factors, both of which have extensively colonized the environment of the NICU. Repeated isolation of these two organisms from the NICU environment proves that some of the neonatal infections may be from the environment itself. Transmission can be stopped by maintaining the sterility of the NICU & hand hygiene among the mothers and health care workers.