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Rapid Identification of Clinical Isolates of Klebsiella Pneumoniae Using MALDI-TOF MS from North India
Bulletin of Pure and Applied Sciences Print version ISSN 0970 0765 Vol.39A (Zoology), No.1, Online version ISSN 2320 3188 January-June 2020: P.194-199 DOI 10.5958/2320-3188.2020.00022.4 Original Research Article Available online at www.bpasjournals.com Rapid Identification of Clinical Isolates of Klebsiella pneumoniae using MALDI-TOF MS from North India 1Sunil Kumar Abstract: 2Zainab Saifi Infections caused by multi-drug resistant (MDR) 3Anil Kumar Sharma Klebsiella pneumoniae are increasing day by day. K. pneumoniae poses a severe public health concern 4 * Sushil Kumar Upadhyay and causes wide array of healthcare associated infections with limited treatment options. The Author’s Affiliation: quick detection of these isolates is of prime 1,2,3,4 Department of Biotechnology, Maharishi importance for the adoption of proper antibiotic Markandeshwar (Deemed to be University), treatment and control measures. Also the mis- Mullana-Ambala, Haryana 133207, India identification using standard lab-based methods is quite common and hence, the clinical *Corresponding author: significance of K. pneumoniae complex members is Dr. Sushil Kumar Upadhyay, inaccurately defined. Here, authors evaluated the Assistant Professor, Department of potential of MALDI-TOF (Matrix-Assisted Laser Biotechnology, Maharishi Markandeshwar Desorption Ionization-Time of Flight) MS (Mass (Deemed to be University), Mullana-Ambala, Spectrometry) to discriminate and quick Haryana 133207, India identification K. pneumoniae complex members. Thus, this study aimed at full MALDI-based E-mail: approach to rapidly detect the K. [email protected] pneumoniae isolates up to species level. We also ORCID: https://orcid.org/0000-0002-1229-4275 explored the antibiotic resistance profile of K. -
Characterization and Antibiotic Sensitivity Profile of Bacteria Isolated from Patients with Respiratory Tract Infections in Bangladesh
Characterization and Antibiotic Sensitivity Profile of Bacteria Isolated from Patients with Respiratory Tract Infections in Bangladesh Shukla Promite1, Sajal K. Saha2, Sunjukta Ahsan1 and Marufa Zerin Akhter1 1Department of Microbiology, University of Dhaka, Dhaka, Bangladesh 2Department of General Practice, Monash University, Building 1, 270 Ferntree Gully Road, Notting Hill VIC 3168, Australia (Received: October 08, 2017; Accepted: December 15, 2017; Published (web): December 23, 2017) ABSTRACT: The study was aimed to characterize bacterial isolates from respiratory tract infections (RTI) and investigate their antibiotic sensitivity profile. Selective media and biochemical tests were used to characterize 40 bacterial isolates. Antibiotic sensitivity testing was conducted using Kirby-Bauer disc diffusion method. About 42.5% (17) RTI patients were infected by Klebsiella pneumoniae, 30% (12) by Escherichia coli and 27.5% (11) by Pseudomonas aeruginosa with no significant gender variation (p-value <0.578). Overall, 47% (out of 20) antibiotics were sensitive, whereas 48% were resistant. Surprisingly, 18% P. aeruginosa and 20% K. pneumoniae were carbapenem-resistant and 4 out of 7 cephalosporin antibiotics were highly resistant irrespective of pathogens. E. coli showed better sensitivity to nitrofurantoin (78%) and levofloxacin (89%), while K. pneumoniae was insensitive to cotrimoxazole (88%), gentamycin (77%) and piperacillin/tazobactam (66%). On the other hand, P. aeruginosa did not respond to P. aeruginosa to nalidixic acid (60%) and ciprofloxacin (60%). This study concludes that nitrofurantoin, levofloxacin, cotrimoxazole, gentamycin and piperacillin/tazobactam antibiotics could be better alternative in treating bacterial RTIs. Key words: Antibiotic sensitivity, bacterial pathogens, RTIs, Bangladesh. INTRODUCTION Antibiotic resistance (AR) is a global public The rise of AR in Bangladesh is probably due to 1 health concern. -
Pasteurella Multocida Isolated from Cattle
Journal of Applied Pharmaceutical Science Vol. 3 (04), pp. 106-110, April, 2013 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2013.3419 ISSN 2231-3354 Antibiotic Susceptibility and Molecular Analysis of Bacterial Pathogen Pasteurella Multocida Isolated from Cattle Azmat Jabeen, Mahrukh Khattak, Shahzad Munir*, Qaiser Jamal, Mubashir Hussain Department of Microbiology, Kohat University of Science and Technology, Kohat, Khyber Pakhtunkhwa, Pakistan. ARTICLE INFO ABSTRACT Article history: Pasteurella multocida is a Gram negative, non motile and coccobacillus bacterium. It has 5 strains i.e. A, B, D, Received on: 01/02/2013 E and F and 16 serotypes (1-16). In present study, we analyzed Pasteurella multocida B: 2 strains, responsible Revised on: 19/02/2013 for Hemorrhagic Septicemia (HS) in cattle, on morphological/microbial, biochemical, molecular level and to Accepted on: 15/03/2013 check the antibiotic sensitivity of the Pasteurella multocida. Microbial analysis showed that while grown on Available online: 27/04/2013 Brain Heart Infusion agar plates and Blood Agar Base Medium, grayish lustrous colonies of Pasteurella multocida were observed. Gram staining showed that Pasteurella multocida are gram negative. Microscopic Key words: observations revealed it to be coccobacillus and it was non- motile. Identification was conducted by Pasteurella multocida, conventional biochemical tests and percentage identification of Analytical Profile Index was 96 %. Antibiotic Hemorrhagic Septicemia, sensitivity with different antibiotics was checked by disk diffusion method and was found resistant to Analytical Profile Index, Augmentin, Amoxicillin and Aztreonam and was more susceptible to Ceftiofur. On molecular level its DNA Antibiotic sensitivity. was extracted and was run with marker having range from 0.5 – 10 kb. -
Identification of Pasteurella Species and Morphologically Similar Organisms
UK Standards for Microbiology Investigations Identification of Pasteurella species and Morphologically Similar Organisms Issued by the Standards Unit, Microbiology Services, PHE Bacteriology – Identification | ID 13 | Issue no: 3 | Issue date: 04.02.15 | Page: 1 of 28 © Crown copyright 2015 Identification of Pasteurella species and Morphologically Similar Organisms Acknowledgments UK Standards for Microbiology Investigations (SMIs) are developed under the auspices of Public Health England (PHE) working in partnership with the National Health Service (NHS), Public Health Wales and with the professional organisations whose logos are displayed below and listed on the website https://www.gov.uk/uk- standards-for-microbiology-investigations-smi-quality-and-consistency-in-clinical- laboratories. SMIs are developed, reviewed and revised by various working groups which are overseen by a steering committee (see https://www.gov.uk/government/groups/standards-for-microbiology-investigations- steering-committee). The contributions of many individuals in clinical, specialist and reference laboratories who have provided information and comments during the development of this document are acknowledged. We are grateful to the Medical Editors for editing the medical content. For further information please contact us at: Standards Unit Microbiology Services Public Health England 61 Colindale Avenue London NW9 5EQ E-mail: [email protected] Website: https://www.gov.uk/uk-standards-for-microbiology-investigations-smi-quality- and-consistency-in-clinical-laboratories UK Standards for Microbiology Investigations are produced in association with: Logos correct at time of publishing. Bacteriology – Identification | ID 13 | Issue no: 3 | Issue date: 04.02.15 | Page: 2 of 28 UK Standards for Microbiology Investigations | Issued by the Standards Unit, Public Health England Identification of Pasteurella species and Morphologically Similar Organisms Contents ACKNOWLEDGMENTS ......................................................................................................... -
Asymptomatic Bacteriuria Amongst the Inhabitants of Okigwe, Imo State Nigeria
Nigerian Journal of Microbiology, Vol. 22(1): 16 30 – 1633 2008 Asymptomatic Bacteriuria Amongst the Inhabitants of Okigwe, Imo State Nigeria *Ugbogu, O.C and Enya, V. N Department of Microbiology, Abia State University, Uturu, Nigeria. Abstract The prevalence of asymptomatic bacteriuria amongst the inhabitants of Okigwe was investigated using culture techniques. The predominant bacteria isolated were Escherichia coli , Staphylococcus aureus , Klebsiella species, Pseudomonas aeruginosa and Proteus species. Out of the 120 urine samples examined 20.8% had asymptomatic bacteriuria. The percentage prevalence was 17.7% and 22.5% for males and females examined respectively. Escherichia coli was the most prevalent occurring in 18.2% of the samples while Klebsiella species and Proteus species that both occurred in 5% of the positive samples were the least. Traders were more affected than students and civil servants. There is need to encourage people to screen for asymptomatic bacteriuria in other to avert the consequences of the subsequent complications. Keywords: bacteriuria, occupation, prevalence, symptom. *Corresponding author; E-mail; [email protected] phone 07084159395 Introduction Materials and methods Normally urine and the urinary tract Population studied : above the entrance to the bladder are essentially The population for this study was a free of microorganisms (Nester et al ., 2004). randomly selected group of 120 aparently Bacteriuria is a condition in which bacteria are healthy individuals that were either students, present in urine. Asymptomatic bacteriuria is traders or civil servants in Okigwe. The study defined as significant bacteriuria when growth of population were of various age groups ranging ≥ 10 5 cfu/ml of freshly voided urine (Umeh et from 16 to 45 years. -
Antibiotic Susceptibility Pattern of Bacterial Uropathogens Isolated
ANTIBIOTIC SUSCEPTIBILITY PATTERN OF BACTERIAL UROPATHOGENS ISOLATED FROM PATIENTS IN NAKURU LEVEL 5 HOSPITAL, KENYA GEORGE TIBI GACHUHI (B.Sc.) I56/CE/20963/2010 A Thesis Submitted in Partial Fulfillment of the Requirements for the Award of the Degree of Master of Science (Microbiology) in the School of Pure and Applied Sciences of Kenyatta University JULY, 2017 ii DECLARATION This thesis is my own original work and has not been presented for the award of a degree in any other university. George Tibi Gachuhi Signature…………………………. Date………………….. Department of Microbiology APPROVAL BY SUPERVISORS This thesis has been submitted for examination with our approval as University supervisors. Dr John Maingi Signature………………………………… Date…………………………………. Department of Microbiology Kenyatta University Dr. Anthony Kebira Signature…………………………………………Date…………………………. Department of Microbiology Kenyatta University iii DEDICATION This research project is dedicated to my beloved wife Salome Tibi and our beloved children Victor, Grace, Martha, Daisy, Caleb and Ann for their support, encouragement and prayer during my time of my study. iv ACKNOWLEDGEMENTS I thank God for His love, guidance and wisdom to be able to successfully complete my research project. I would like to appreciate a number of people who helped me during the preparation and writing of this research project especially Dr. John Maingi and Dr. Anthony Kebira of Kenyatta University, Department of Microbiology. Thank you for your advice and guidance. They have not only been outstanding supervisors, but credible role models. Their guidance, support and encouragement right from the proposal development to the final report would not have been successfully completed without them. I am greatly indebted to the staff at Nakuru Level 5 Hospital especially Dr. -
MALDI-TOF MS IDENTIFICATION of CITROBACTER YOUNGAE ISOLATED from FOOD Dubravka S
DOI: 10.5937/FFR1802107M UDK 579.84:616.98]:637.3 Short communication MALDI-TOF MS IDENTIFICATION OF CITROBACTER YOUNGAE ISOLATED FROM FOOD Dubravka S. Milanov*1, Milan D. Đilas2, Jelena М. Babić1, Bojana Z. Prunić1, Maja Ј. Velhner1 1 Scientific Veterinary Institute ‘Novi Sad’, Rumenački put 20, 21000 Novi Sad, Serbia 2 Institute of Public Health of Vojvodina, Futoška 121, Novi Sad, Serbia *Corresponding author Tel.: +381 21 4895 346 E-mail: [email protected] ABSTRACT: Salmonella-like bacteria, such as certain species of the Citrobacter genus are sometimes isolated from food and feed. By testing their biochemical and, partially, serological characteristics (according to the regulations given in the ISO 6579-1:2017 Standard) it is impossible to completely reject their affiliation to the Salmonella genus. In addition to this, the food microbiology laboratories usually lack diagnostic tools and procedures for precise identification of these isolates to the species level. Thus, it is considered useful to describe the identification of Citrobacter youngae species using MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry). Besides differential diagnostic criteria for distinguishing between Salmonella spp. and Citrobacter youngae, in this communication the relevant data on Citrobacter spp., the emerging opportunistic pathogenic member of Enterobacteriaceae family were presented. Key words: Salmonella-like bacteria, cheese A REPORT ON A LABORATORY CASE A sample of soft, medium-fat cheese accordance with the ISO 6579-1:2017 sprinkled with spices, produced in a small Standard (Table 1), plus using the methyl- scale plant, was examined for the pre- red and citrate utilization tests, which were sence of Salmonella spp. -
Subsurface Endospore-Forming Bacteria Possess Bio-Sealant
www.nature.com/scientificreports OPEN Subsurface Endospore-Forming Bacteria Possess Bio-Sealant Properties Received: 14 July 2017 Sreenivasulu Basha1, Lakshman Kumar Lingamgunta2, Jayakumar Kannali1, Swarna Kumari Accepted: 5 April 2018 Gajula1, Ramesh Bandikari2,3, Sreenivasulu Dasari2, Veena Dalavai1, Paramageetham Published: xx xx xxxx Chinthala1, Prasada Babu Gundala1, Peera Kutagolla4 & Vinodh Kumar Balaji5 Concrete is a strong and fairly inexpensive building substance, but has several disadvantages like cracking that allows corrosion, thus reducing its lifespan. To mitigate these complications, long-lasting microbial self-healing cement is an alternative that is eco-friendly and also actively repairs cracks. The present paper describes the detailed experimental investigation on compressive strength of cement mortars, mixed with six alkaliphilic bacteria, isolated from subsurface mica mines of high alkalinity. The experiments showed that the addition of alkaliphilic isolates at diferent cell concentrations (104 and 106 cells/ml) enhanced the compressive strength of cement mortar, because the rapid growth of bacteria at high alkalinity precipitates calcite crystals that lead to flling of pores and densifying the concrete mix. Thus, Bacillus subtilis (SVUNM4) showed the highest compressive strength (28.61%) of cement mortar at 104 cells/ml compared to those of other fve alkaliphilic isolates (Brevibacillus sp., SVUNM15-22.1%; P. dendritiformis, SVUNM11-19.9%; B. methylotrophicus, SVUNM9-16%; B. licheniformis, SVUNM14- 12.7% and S. maltophilia, SVUNM13-9.6%) and controlled cement mortar as well. This method resulted in the flling of cracks in concrete with calcite (CaCO3), which was observed by scanning electron microscopy (SEM). Our results showed that the alkaliphilic bacterial isolates used in the study are efective in self-healing and repair of concrete cracks. -
Bio201lab12.Exp.15.1
Professor Diane Hilker I. Exp. 15: Physiology of Bacteria Purpose: To examine specific enzymatic activities of microbes that are frequently used to identify bacterial species. Inoculated last lab: ◦ E.coli, Enterobacter, Proteus: Gram Neg rods ◦ Bacillus sp.: Gram Pos. rod Phenol Red Dextrose Broth (PRDB): does the microbe ferment glucose or dextrose? ◦ Yellow with gas: + ◦ Yellow without gas: - ◦ Red with or without gas: - + - - ◦ Yellow/red with or without gas: +/- Phenol Red Lactose Broth (PRLB): does the microbe ferment lactose? ◦ Yellow with gas: + ◦ Yellow without gas: - ◦ Red with or without gas: - + - - ◦ Yellow/red with or without gas: +/- Nitrate Broth: Does the microbe produce an enzyme called nitratase? Nitrate Nitrite Nitratase ADD: 2-3 drops Nitrate A Mix; look for color 2-3 drops Nitrate B development in 30sec. Nitrate Broth RESULTS: Peach/pink: + Not peach/pink: - - + Tryptone Broth: Does the microbe produce an enzyme called tryptophanase? Tryptophan Indole Tryptophanase ADD: 10-12 drops of Kovacs Reagent Mix; look immediately for the reaction Tryptone Broth RESULTS: Maroon top layer: + No maroon top layer: - Methyl Red- Voges Proskauer Broth (MRVP) First divide the tube in half using a Pasteur pipette. Transfer ½ of broth to a 2nd empty glass tube. Cap both tubes. One tube you will perform the Methyl Red Test and the 2nd the Voges Proskauer Test Methyl Red Test: Does the microbe produce a large amount of acid end product from glucose fermentation? Glucose pH below 4.4 ADD: 4 drops of Methyl Red Reagent Mix; look immediately for the reaction Methyl Red Test RESULTS: Pink: + Not Pink: - Voges Proskauer Test: Does the microbe produce a compound called acetoin during glucose fermentation? Glucose Acetoin ADD: 18 drops of Barritts A Reagent 18 drops of Barritts B Reagent Mix tube well & let stand for 10 minutes. -
Asian Journal of Medical and Biological Research Detection Of
Asian J. Med. Biol. Res. 2016, 2 (4), 656-663; doi: 10.3329/ajmbr.v2i4.31011 Asian Journal of Medical and Biological Research ISSN 2411-4472 (Print) 2412-5571 (Online) www.ebupress.com/journal/ajmbr Article Detection of bacterial species from clinical mastitis in dairy cows at Nilphamari district and their antibiogram studies Md. Titon mia 1, Md. Khaled hossain 2, Nazmi Ara Rumi3*, Md. Shajedur Rahman 4, Md. Shahin Mahmud 5 and Monoranjan Das 6 1Department of Microbiology, Hajee Mohammad Danesh Science &Technology University, Dinajpur, Bangladesh 2Department of Microbiology, Hajee Mohammad Danesh Science &Technology University, Dinajpur, Bangladesh 3Department of Microbiology, Hajee Mohammad Danesh Science &Technology University, Dinajpur, Bangladesh 4Department of Medicine, Surgery and Obstetrics, Hajee Mohammad Danesh Science& Technology University, Dinajpur, Bangladesh 5Department of Microbiology, Hajee Mohammad Danesh Science &Technology University, Dinajpur, Bangladesh 6Department of Microbiology, Hajee Mohammad Danesh Science &Technology University, Dinajpur, Bangladesh *Corresponding author: Nazmi Ara Rumi, Lecturer, Department of Microbiology, Hajee Mohammad Danesh Science & Technology University, Dinajpur, Bangladesh. Phone: +8801774410044; E-mail: [email protected] Received: 17 November 2016/Accepted: 20 December 2016/ Published: 29 December 2016 Abstract: The present study was conducted on the rural dairy cows to detect the bacterial species from clinical mastitis in dairy cows with their antibiogram studies during the period from January 2015 to June 2015. For this purpose two upazilla were selected under the Nilphamari district. On the basis of morphology, staining, cultural and biochemical characteristics, the isolated organisms were classified as, Staphylococcus spp., Streptococcus spp., E. coli, and Bacillus spp. .For this study, a total of 48 samples were collected from affected mastitis cows. -
Identification of Bacteria Associated with Lower Respiratory Tract Among Patients Attending in General Hospital Dr
Online - 2455-3891 Vol 11, Issue 9, 2018 Print - 0974-2441 Research Article IDENTIFICATION OF BACTERIA ASSOCIATED WITH LOWER RESPIRATORY TRACT AMONG PATIENTS ATTENDING IN GENERAL HOSPITAL DR. PIRNGADI MEDAN JEFRI NALDI1*, ROSIDAH ROSIDAH1, RICKE LOESNIHARI2 1Department of Pharmacology, Faculty of Pharmacy, University of Sumatera Utara, Medan, Indonesia. 2Department of Clinical Pathology, Faculty of Medicine, University of Sumatera Utara, Medan, Indonesia. Email: [email protected] Received: 16 April 2018, Revised and Accepted: 18 May 2018 ABSTRACT Objective: The aim of this study was to establish cases of bacteria associated with lower respiratory tract infections (LRTIs) in General Hospital Dr. Pirngadi Medan with the view to identify the bacteria well as to assess their antibiotics susceptibility. Methods: An observation and prospective study were analyzed to patients with lower respiratory tract infection in respiratory and internal wards Dr. Pirngadi Hospital Medan. For the identification method of bacteria was using optochin test, catalase test, Mannitol salt agar, coagulase, indole, methyl red, Voges–Proskauer test, citrate, urease, and Triple Sugar Iron test. Results: The bacteria that cause lower respiratory tract infection were Pseudomonas aeruginosa (25.53%), Klebsiella pneumoniae (17.02%), Proteus sp. (14.89%), Staphylococcus aureus (12.77%), Escherichia coli (10.64%), Streptococcus pneumoniae (10.64%), and Haemophilus influenzae (8.51%). The result of sensitivity testing showed that gentamicin is more sensitive and the resistant antibiotic is chloramphenicol among all the tested antibiotics. Conclusion: P. aeruginosa was the most common isolated from LRTIs patients with gentamicin is the most sensitive antibiotics while chloramphenicol is the most resistant antibiotics among all the tested antibiotics. Keywords: Bacteria, Antibiotic, Lower respiratory tract infection, Susceptibility. -
Characterization of Klebsiella Isolates by Matrix-Assisted Laser Desorption
Turkish Journal of Medical Sciences Turk J Med Sci (2015) 45: 1335-1344 http://journals.tubitak.gov.tr/medical/ © TÜBİTAK Research Article doi:10.3906/sag-1401-32 Characterization of Klebsiella isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and determination of antimicrobial resistance with VITEK 2 advanced expert system (AES) 1 1, 2 Alper KARAGÖZ , Sümeyra ACAR *, Hanifi KÖRKOCA 1 Public Health Institution of Turkey, Molecular Microbiology Research and Application Laboratory, Ankara, Turkey 2 Department of Nursing, School of Health, Muş Alparslan University, Muş, Turkey Received: 08.01.2014 Accepted/Published Online: 09.07.2014 Printed: 31.12.2015 Background/aim: The purpose of the study was to evaluate the performance of the VITEK mass spectrometry (MS) (bioMérieux, France) system for the identification of Klebsiella spp. isolated from different sources. Moreover, while assessing the ability of the VITEK 2 automated expert system (AES) to recognize antimicrobial resistance patterns, the researchers have extended the study to compare VITEK 2 with the routine antimicrobial susceptibility testing method. Materials and methods: This study tested 51 Klebsiella spp. isolates that were isolated from environmental examples and clinical examples. Results of conventional methods and the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS were compared. Then, any differing results were compared against a reference 16S rRNA gene sequence, and when indicated, a recA sequencing analysis was done. Results: VITEK MS correctly identified 100% of the Klebsiella spp. isolates. There were two K. oxytoca isolates incorrectly identified to the species level with conventional methods according to the 16S rRNA gene sequencing analysis.