20 IJSR - INTERNATIONAL JOURNAL OF SCIENTIFIC RESEARCH Volume : 5 | Issue : 12 | December 2016 • ISSN No 2277 - 8179 | IF : 3.508 | IC Value : 78.46 Original Research Paper Microbiology * Pradnya Atmaram Jadhav Department of Microbiology, Bharati Vidyapeeth Deemed University Medical College and Hospital, Sangli- 416414, Maharashtra, India, * Corresponding author Shahriar B. Roushani Department of Microbiology, Pravara Medical College and Tertiary Care Hospital- Loni, Ahmednagar, Maharashtra, India HiCrome UTI Agar : An Alternative To Conventional Media For Presumptive Identification And Isolation Of Uropathogens KEYWORDS : Uropathogens, Hi- Crome UTI agar, CLED agar, BA and MA agar ABSTRACT The increase in resistance of uropathogens to antimicrobial agents, especially in hospitalized patients, demands rapid identification of the pathogen. Current detection methods including culture and identification are time-consuming. The chromogenic media offers simultaneous presumptive identification of gram-positive and gram-negative bacteria and yeasts on a single medium by means of distinct colony colours produced by reactions of genus- or species-specific enzymes with a suitable chromogenic substrate. The present study was undertaken to validate the usefulness of HiCrome UTI agar as a primary urine culture medium for its rate of isolation and presumptive identification of uropathogens in comparison to CLED agar, BA and MA agar. This study included 1500 midstream and/or catheter-catch urine samples from clinically suspected UTI patients of different age and sex groups. HiCrome UTI agar, MacConkey agar , Blood agar and CLED agar media were obtained as a dehydrated powder from HiMedia laboratories. Urine culture was done on above said media and presumptive identification of bacterial growth was done Out of total 1500 urine samples,552 urine samples showed significant growth of a single isolate and 13 urine samples showed mixed growth of two organisms and 50% samples were sterile. Among all the bacterial isolates E.coli were maximum in number (n=195), followed by Klebsiella spp. (n=81).Staphylococcus aureus and Candida spp. constitute 12.97 % and 7.61% of the total urinary isolates, respectively. Maximum number of urinary isolates was presumptively identified on Hicrome UTI agar (88.06 %) and there is no significant difference in the rate of presumptively identification from CLED agar and Blood agar and MacConkey agar in combination (62.97% and 61.24, respectively). Presumptively identification of mixed growth of organisms on Hicrome UTI agar was 100%, but CLED agar, Blood agar and MacConkey agar in combination failed to presumptively identify 100% mixed growth of organisms. The overall findings of this study suggests that though expensive, chromogenic media like HiCrome UTI Agar media, is an acceptable alternative to traditional media for the isolation of urinary pathogens. Introduction: Urinary tract infection (UTI) is the most common type of infection and continues to be a major health problem. The increase in resistance of microorganisms to antimicrobial agents, especially in hospitalized patients, demands rapid identification of the pathogen. Early information enables the selection of the appropriate antibiotics prior to the re- sults of standard susceptibility tests and may thereby pre- vent outbreaks. (1) For many years Blood, Cystine lactose electrolyte-deficient, and MacConkey agars have been used for the detection of urinary tract pathogens, as well as for the differentiation of a few of them. (2) These current detection methods are time-consuming, most typically consisting of 2 to 3 days of culture. (1) Therefore any new method or medium with the ability to streamline urine culture processing in a meaning- ful way should be welcomed. The aim of the microbiology laboratory is to reduce mor- bidity through accurate identification of pathogens with appropriate antimicrobial sensitivity testing in short turna- round time. (3) In the last few years several chromogenic media have been developed and commercialized, allowing for more specific direct differentiation of microorganisms on primary plates. The chromogenic agar offers simultaneous presumptive identification of gram-positive and gram-negative bacteria and yeasts on a single medium by means of distinct colony colours produced by reactions of genus- or species-specific enzymes with a suitable chromogenic substrate. (2) The present study was undertaken to validate the useful- ness of HiCrome UTI agar as a primary urine culture me- dium for its rate of isolation and presumptive identification of uropathogens in comparison to CLED agar, Blood and Mac Conkey agar in a Pravara Tertiary Care Hospital, Loni, Ahamednagar. Materials and Methods: The protocol was approved by Institutional Ethical Com- miee, Pravara Medical College and Hospital, Loni and the study included 1500 midstream and/or catheter-catch urine samples from clinically suspected UTI patients of dif- ferent age and sex groups aended either at the outpatient department or admied in Pravara Tertiary Care Hospital, Loni, from March 2011 to August 2012. A detail medical history of the patient was taken and the data was recorded. Preparation of media: HiCrome UTI agar, MacConkey agar , Blood agar and CLED agar media were obtained as a dehydrated powder from HiMedia laboratories (HiMedia Laboratories Pvt. Ltd. Mumbai-400086, India). All culture petri plates were prepared in laboratory by following man- ufacturer’s instructions and recommendations. Quality control: Each batch of medium used in this trial was tested for sterility, biochemicals and chromogenic reactions with American Type Culture Collection (ATCC) strains. ATCC Control strains: Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Proteus mirabilis ATCC 12453, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae 13883, and Candida albi- cans ATCC 10231 were used for quality control. (2) Urine culture was performed for samples that showed pus cells ≥ 5/ HPF on microscopy of a centrifuged deposit of urine (hicrome 2). All the urine samples were inoculated aseptically on HiCrome UTI agar, CLED agar, Blood agar and MacConkey agar media with a calibrated loop and were incubated aerobically at 37 0 C for 18-20 hours. Criteria for significant bacteriuria (4) : 1. Presence of >10 5 cfu/ml of non-coliforms or >10 2 cfu/ml of coliforms in a symptomatic woman. 2. Presence of >10 3 cfu/ml of bacteria in a symptomatic man. 3. Growth of two different organisms from possible uropathogens at a con- centration of 10 4 cfu/ml.