Short Communication DOI: 10.18231/2394-5478.2018.0060 Indian Journal of Microbiology Research, April-June, 2018;5(2):288-289 288 Identification of emerging Chryseobacterium indologenes sepsis using automated identification system, Matrix-assisted Laser Desorption Ionization-time-of-flight Mass Spectrometry (MALDI-TOF) and Pulsed-field Gel Electrophoresis (PFGE) typing Awadhesh Kumar 1 , Chinmoy Sahu 2,* , Avinash Singh 3 , Kashi Nath Prasad 4 , Tapan K. N. Dhole 5 1 Lecturer, 2 Assistant Professor, 3 SRF, 4,5 Professor, Dept. of Microbiology, 1 Mahamaya Rajkiya Allopathic Medical College, Ambedkar Nagar, Uttar Pradesh, 2-5 Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, India *Corresponding Author: Email: sahu.chinmoy@gmail.com Abstract Chryseobacterium indologenes is a non-motile, catalase-positive, oxidase-positive, indole-positive, non-glucose-fermenting Gram- negative bacillus that produces yellow coloured colonies on routine culture media. As common environmental saprobe, it is a known colonizer of inanimate objects in the hospital environment. Chryseobacterium indologenes is an emerging .pathogen of nosocomial origin in background of increased use of higher antibiotics. In the present study, positive blood cultures bottled from critically ill patients suspected of nosocomial sepsis were inoculated on routine bacteriological media for a period of six months. The identity and antibiotic sensitivity of Chryseobacterium indologenes were confirmed by Phoenix automated system® followed by automated MALDI-TOF system and Pulsed Field Gel Electrophoresis (PFGE) typing. All the patients had underlying morbidities (7 out of 7 patients). Phoenix automated system and MALDI-TOF system correctly identified the bacterial isolates. We reiterate the fact that levofloxacin should be the first choice of antibiotic in Chyrseobacterium indologens infections followed by cefoperazone + sulbactam. All the isolates were of different clonality as determined by PFGE. To our knowledge, this is the first report on the identification of Chrysobacterium indologenes isolates using automated system, MALDI-TOF and PFGE in India. So, in ICUs of tertiary care centers, where higher antibiotics are used, this bacteria can be an important cause of nosocomial bacteremia. Keywords: Chryseobacterium indologenes, sepsis, MALDI-TOF, PFGE, Phoenix automated system. Introduction Chryseobacterium indologenes is a rare cause of bacterial infection in immunosuppressed patients and belongs to group Chryseobacteria. 1 Chryseobacteria are a group of nonmotile, catalase-positive, oxidase- positive, indole-positive, non-glucose-fermenting, gram negative bacilli. 2 The genus Chryseobacterium includes six species that were previously designated the genus Flavobacterium. Chryseobacterium gleum and Chryseobacterium indologenes, previously known as Flavobacterium CDC group IIb, have been clearly differentiated by DNA-DNA homology and eight phenotypic characteristics. 2 Infections caused by C. indologenes are generally associated with indwelling devices and because it is an environmental bacterium and hence, able to cause nosocomial infections. 3 It has been implicated in nosocomial bacteremia, Urinary Tract Infections, pneumonia etc. 3-6 Nearly half of the published research reports refer to nosocomial infections, and a vast majority of patients had underlying immuno- compromised conditions. 4-10 Studies indicated that the infections caused by C. indologenes increased with the increasing use of higher antibiotics like colistin especially in critically ill patients in ICU. 9 However, the treatment against these pathogens is difficult because they are resistant to many antibiotics especially colistin and carbapenems. 6 They are also reported to be sensitive to Levofloxacin. 6,11,12 To date, the clinical significance of C. indologenes has not been fully defined. Few cases have been reported about the bacterium in world. Therefore, the present study was undertaken to identify the C. indologenes isolates using automated system, MALDI-TOF and PFGE. Materials and Methods The present study was conducted in the Microbiology Department, Sanjay Gandhi Post Graduate Institute of Medical Sciences (SGPGIMS), Lucknow, a tertiary care institute in the Northern India. Blood samples were collected from ill patients suspected of clinical sepsis within a period of six months (July 2015 to December 2015). Around 10 ml of blood were collected aseptically and put in BD Bactec plus aerobic and anaerobic vial. Afterwards, the bottles were incubated at 37ºC aerobically and anaerobically, respectively for maximum five days. Positive blood culture bottles were inoculated on routine bacteriological media like blood agar and MacConkey agar and incubated overnight. Gram negative, oxidase-positive, non-fermenting, yellow pigment producing colonies were appeared and suspected of Chryseobacteria. A characteristic of Chryseobacteria was the development of red color upon addition of 10% Potassium Hydroxide solution to the bacterial culture and was due to the production of flexirubin pigment. The identity and