Microbiology Examination
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Meningitis Manual Text
Laboratory Methods for the Diagnosis of MENINGITIS Caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae Centers for Disease Control and Prevention August, 1998 Laboratory Methods for the Diagnosis of Meningitis Caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae Table of Contents Introduction………………………………………………………………………………… 1 Acknowledgments ……………………………………………………………………….. 2 I. Epidemiology of Meningitis Caused by Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae,…………………………………………… 3 II. General Considerations ......................................................................................................... 5 A. Record Keeping ................................................................................................................... 5 III. Collection and Transport of Clinical Specimens ................................................................... 6 A. Collection of Cerebrospinal Fluid (CSF)............................................................................... 6 A1. Lumbar Puncture ................................................................................................... 6 B. Collection of Blood .............................................................................................................. 7 B1. Precautions ............................................................................................................ 7 B2. Sensitivity of Blood Cultures ................................................................................ -
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. -
Multicellular Oxidant Defense in Unicellular Organisms MUCHOU MA and JOHN W
Proc. Natl. Acad. Sci. USA Vol. 89, pp. 7924-7928, September 1992 Microbiology Multicellular oxidant defense in unicellular organisms MUCHOU MA AND JOHN W. EATON* Division of Experimental Pathology, Department of Pathology and Laboratory Medicine, Albany Medical College, A-81, 47 New Scotland Avenue, Albany, NY 12208 Communicated by David W. Talmage, May 8, 1992 ABSTRACT Although catalase is thought to be a major MATERIALS AND METHODS defense against hydrogen peroxide (H202), the catalase activity Reagents. Brain heart infusion broth, Todd-Hewitt broth, within individual Escherichia coil fails to protect against ex- Lennox L agar (LB agar), and Bactoagar were obtained from ogenous H202. Contrary to earlier reports, we find that dilute GIBCO/BRL. The bicinchoninic acid protein microassay suspensions, of wild-type and catalase-deficient E. colt are was from Pierce. All other enzymes and chemicals were identical in their sensitivity to H202, perhaps because even purchased from Sigma. wild-type, catalase-positive E. colU cannot maintain an inter- Bacterial Strains and Culture Conditions. A catalase- nal/externail concentration gradient of this highly diffusible deficient mutant strain of E. coli K-12 [UM1, hereafter, oxidant. However, concentrated suspensions or colonies of cat(-)] and its parent wild-type [CSH7, hereafter cat(+)] (17) catalase-positive E. colt do preferentially survive H202 chal- were provided by P. C. Loewen (University ofManitoba). E. lenge and can even cross-protect adjacent catalase-deficient coli were grown statically in brain heart infusion broth or M9 organisms. Furthermore, high-density catalase-positive-but minimal salts medium supplemented with 10 mM glucose not catalase-negative-E. colt can survive and multiply in the (M9/glucose) (25) at 370C in room air overnight (18-20 hr) to presence of competitive, peroxide-generating streptococci. -
Growth Characteristics of Escherichia Coli and Staphylococcus Aureus Bacteria on Alternative Medium Leaves of Lamtoro (Leucaena Leucocephala)
Journal of Xi'an University of Architecture & Technology ISSN No : 1006-7930 Growth Characteristics of Escherichia coli and Staphylococcus aureus Bacteria on Alternative Medium Leaves of Lamtoro (Leucaena leucocephala) Meidawati Suswandari*, Department of Primary School, Faculty of Teacher Training and Education, Universitas Veteran Bangun Nusantara, Sukoharjo, Indonesia Lamtoro leaf has a high protein content. The protein content is very suitable for bacterial growth. Because of the high cost of bacterial growth media for educational and research institutions, lamtoro leaves can be used as an alternative medium for bacterial growth in general. The purpose of this study was to determine the potential of lamtoro leaf as an alternative medium for bacterial growth in general. This research is descriptive. Alternative mediums of lamtoro leaf were tested for the growth of Escherichia coli and Staphylococcus aureus. Escherichia coli bacteria grow on three alternative medium plates. After final identification, there are Escherichia coli bacteria. Whereas the Staphylococcus aureus bacterium did not grow on seven plates of alternative medium despite being incubated for 48 hours. Lamtoro leaf has less potential as an alternative medium for bacterial growth in general. The lamtoro leaf medium can only be used as a growth medium for gram-negative bacteria. While the growth of gram-positive bacteria there is no growth due to the presence of active substances in lamtoro leaves. Key words: Leaves of Lamtoro, Alternative Media, Escherichia coli, Staphylococcus aureus Introduction Bacteria are single-celled creatures that are very small or microscopic. Hans Christian Gram divides bacteria based on the characteristics of cell walls through the Gram staining system, namely Gram Positive and Gram Negative bacteria (Elferia, et al, 1996; Elliot, 2013; Harvey, 2001; Clausen, Gildberg, and Raa, 1985). -
“Isolation of Antibiotic Producing Actinomycetes from Soil”
© 2020 JETIR May 2020, Volume 7, Issue 5 www.jetir.org (ISSN-2349-5162) “ISOLATION OF ANTIBIOTIC PRODUCING ACTINOMYCETES FROM SOIL” *Sneha khadse1Priyanka Titirmare2 1) Project assistant at CSIR NEERI Nagpur, 2) Kamla Nehru Mahavidyalaya, Nagpur. ABSTRACT The present study deals with the isolation, production, purification and optimization of antibiotics from soil. The soil samples were collected from various places. Actinomycetes strains were isolated in specific medium using starch casein agar medium. The actinomycetes were screened with regard to potential against gram positive and gram negative bacteria. The purified actinomycetes strains were performed in biochemical tests. Gram staining, sugars fermentation, HS production, urease production, MR-VP test, indole production, citrate 2 utilization. Fermentation was carried out with starch casein broth and identified strain of actinomyces as inoculums. Antimicrobial activity of antibiotics was detected on Mueller Hinton Agar Media against E.coli. The purification of antibiotics was done with the help of activated charcoal. During the experiment a modified fermentation medium was formulated and activity of antibiotic produced in this medium was detected against E.coli. The isolated bacteria such as actinomycetesobtained from the soil. INTRODUCTION Antibiotics are complex chemical secondary metabolites, which are produced by microorganisms & act against other microorganisms. The term antibiotics had its origin from Greek word anti-against+biotikos =”fit for life”. Antibiotics are the best known products of actinomycetes. The best known genus of actinomycetes; Streptomyces (Order Actinomycetales, family Streptomycetaceae are gram-positive, filamentous bacteria that are ubiquitous in soil and produce the majority (>70%) of known antibiotics (Tanaka and Omura, 1990). The isolation & characterization of Actinomycetes were performed in different biochemical methods. -
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 ......................................................................................................... -
Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation
City University of New York (CUNY) CUNY Academic Works Open Educational Resources Queensborough Community College 2016 Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation Joan Petersen CUNY Queensborough Community College Susan McLaughlin CUNY Queensborough Community College How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/qb_oers/16 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation By Dr. Susan McLaughlin & Dr. Joan Petersen Queensborough Community College Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation Table of Contents Preface………………………………………………………………………………………i Acknowledgments…………………………………………………………………………..ii Microbiology Lab Safety Instructions…………………………………………………...... iii Lab 1. Introduction to Microscopy and Diversity of Cell Types……………………......... 1 Lab 2. Introduction to Aseptic Techniques and Growth Media………………………...... 19 Lab 3. Preparation of Bacterial Smears and Introduction to Staining…………………...... 37 Lab 4. Acid fast and Endospore Staining……………………………………………......... 49 Lab 5. Metabolic Activities of Bacteria…………………………………………….…....... 59 Lab 6. Dichotomous Keys……………………………………………………………......... 77 Lab 7. The Effect of Physical Factors on Microbial Growth……………………………... 85 Lab 8. Chemical Control of Microbial Growth—Disinfectants and Antibiotics…………. 99 Lab 9. The Microbiology of Milk and Food………………………………………………. 111 Lab 10. The Eukaryotes………………………………………………………………........ 123 Lab 11. Clinical Microbiology I; Anaerobic pathogens; Vectors of Infectious Disease….. 141 Lab 12. Clinical Microbiology II—Immunology and the Biolog System………………… 153 Lab 13. Putting it all Together: Case Studies in Microbiology…………………………… 163 Appendix I. -
Biochemical Media Casitone Broth-White Cap with Black Circle. Loop Transfer
Biochemical Media Casitone Broth-white cap with black circle. Loop transfer. (Use care tops may be very loose) (Do not put caps on the wrong tubes during the inoculation procedures). Can the organism split tryptophan into indole, pyruvic acid and ammonia? Test for the presence of indole (unique to this process) by adding 5 drops of Kovacs Test Reagent. If indole is present, a ruby red color will form at the top of the test tube. The media name for casitone comes from the word casein. Casein is a milk protein rich in the amino acid tryptophan. Result sheet recorded with a “pos” or “neg” Phenol Red Carbohydrate Fermentation Broths (Phenol Reds) Red capped broth with durham tube-Phenol Red with Lactose Clear capped broth with durham tube-Phenol Red with Glucose Yellow capped broth with durham tube-Phenol Red with Sucrose Blue capped broth with durham tube-Phenol Red with Maltose Green capped broth with durham tube-Phenol Red with Mannitol All tubes are inoculated with loop transfers. Can the organism ferment the carbohydrates listed above? Each comes with a separate sugar, a durham tube for gas collection and the acid-base indicator called phenolphthalein. Fermentation will produce acidity and sometimes gas. Gas alone is not produced by the fermentation process. Phenolphthalein will turn yellow in the presence of acid, red in an alkaline environment. Gas production can be observed by the presence of air bubbles captured in the durham tube. For each result both an acid and gas response should be indicated. For a large amount of acid (yellow) use “A”. -
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. -
Canada 57-2 Fisheries Peches L+I and Oceans Et Oceans STANDARD PROCEDURES PROCEDURES POUR for BACTERIOLOGICAL ANALYSES ANALYSIS BACTERIOLOGIQUES
/0.58:29 Fisheries Peches l+I and Oceans et Oceans Inspection Services de Services )'inspection Standard Procedures pour Procedures for analyses Bacteriological bacteriologiques Analysis ( /v/fJ,rJ Ul.LS IX 54.3 Canada 57-2 Fisheries Peches l+I and Oceans et Oceans STANDARD PROCEDURES PROCEDURES POUR FOR BACTERIOLOGICAL ANALYSES ANALYSIS BACTERIOLOGIQUES Record of Amendments amend. no chapter date of amend. inserted by insertion date 1 5 +Qn1"' E 31 - ns- X'S C? c l~ Le.Lle.J/ gs - Dtc C; ··7 ~ />f» c ~ e:. ~.J J-~ \ s .. o sfft8 /vf:-l~ fb. ~-o 1~1r . Government Gouvernement of Canada du Canada IDate ••• Canadian Food Agence canadienne 1 15£05£98 Inspection Agency d'inspection des aliments STANDARD PROCEDURES PROCEDURES POUR FOR BACTERIOLOGICAL ANALYSES ANALYSIS BACTERIOLOGIQUES Bulletin TO: All Holders of the Standard Procedures for Bacteriological Analysis Manual SUBJECT: MODIFICATIONS TO THE PROCEDURES Attached are modifications to various Chapters and Appendices of the Bacteriological Analysis manual. The changes have been incorporated in this bulletin format in order to provide the updates to manual holders as quickly as possible and also due to the uncertainty as to whether the manual will continue to be used as a bacteriological procedures reference by the Agency. The changes to the respective sections have been incorporated on separate pages so the pertinent page(s) may be inserted with the relevant chapter/appendix. If required, these modifications will be incorporated in the manual at a later date. Cam Prince Director Fish,