As per the latest CBME Guidelines | Competency Based Undergraduate Curriculum for the Indian Medical Graduate 9 789354 660689 Under the new Competency Based Undergraduate Curriculum for the Indian Medical Graduate, the MBBS students in Phase II require training different from the traditional and conventional methods to a system based teaching module. The primary aim of this practical manual is to assist MBBS students to spend quality time in performing relevant experiments under supervision and use this manual as an easy guide to learn experimental procedures. The content of the book minimizes the burden on the students and gives them the opportunity to have ample time in learning. Under the current curriculum the student is expected to maker relevant observations and record them too. It also contains depiction of important related images and figures which gives the student a comprehensive understanding of the subject and is useful in preparing for the viva voce examination. The practical manual will not only guide the students on relevant topics but their competency will also be simultaneously assessed by the teachers on daily basis. Rumpa Saha MBBS, MD (Microbiology) is currently Professor, Department of Microbiology, University College of Medical Sciences and GTB Hospital, Delhi. She has been associated with this hospital since 2002. After completing her MBBS from Calcutta National Medical College, Calcutta University, she did her MD, Microbiology from PGIMER, Chandigarh. She has more than 16 years of teaching experience to PGs, UGs, nursing and MLT students. Her area of interest includes enteric diseases, parasitology and STIs. Besides conducting and supervising research in microbiology, she has to her credit 73 papers published in indexed national and international scientific journals of repute and has contributed chapters for Osteoarticular Tuberculosis, CBS Publishers & Distributors; Rational Use of Antimicrobials: Indian Perspectives, IMA Publications; Diploma in MLT Microbiology, IMA Publications, and has also edited the last two books. She has been examiner for MBBS, MD (Microbiology) and BDS of a number of universities. She is an active member of several scientific societies and has also delivered guest lectures and chaired sessions at a number of conferences / symposia in India. Shukla Das MD, DNB, MNANS is currently Director-Professor, Department of Microbiology, University College of Medical Sciences and Guru Teg Bahadur Hospital, Delhi; former Assistant Director, National Centre for Disease Control, has to her credit more than 120 publications and 25 years of experience of teaching microbiology to medical undergraduates, postgraduates, nursing and dental students. She has guided several PhD students registered under Faculty Medical Sciences (FMS), Delhi University. Besides conducting teaching programs, she has completed 10 government funded projects in autoimmunity, diarrhoeal diseases and fungal infections. She has published 120 peer-reviewed and indexed papers and written chapters in 6 textbooks. She has been examiner for BDS, MBBS, MD and PhD examinations at several universities. She has been invited to deliver scientific talks at various national and international conferences. She is also the Joint Secretary, Indian Society for Human and Animal Mycology (SIHAM). She has been awarded several best paper poster/oral presentations of which the Satish Chandra best research paper award by IAMM, Delhi Chapter, in 2016, and Dr AM Saoji and Dr SR Sengupta memorial award at national conference MICROCON 2014 were the most coveted. MICROBIOLOGY MICROBIOLOGY Practical Manual Practical Manual Second Edition MICROBIOLOGY MICROBIOLOGY Practical Manual Practical Manual MICROBIOLOGY MICROBIOLOGY Practical Manual Practical Manual Second Edition Second Edition ISBN: 978-93-54660-68-9 4819/XI, Prahlad Street, 24 Ansari Road, Daryaganj, New Delhi 110 002, India E-mail: [email protected], [email protected]; Website: www.cbspd.com New Delhi Bengaluru Chennai Kochi Kolkata Lucknow Mumbai | | | | | | Hyderabad Jharkhand Nagpur Patna Pune Uttarakhand | | | | | |
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As per the latest CBME Guidelines |Competency Based Undergraduate Curriculum for the Indian Medical Graduate
9 789354 660689
SahaDas
Under the new Competency Based Undergraduate Curriculum for the Indian Medical Graduate, the MBBS students in Phase II require training different from the traditional and conventional methods to a system based teaching module. The primary aim of this practical manual is to assist MBBS students to spend quality time in performing relevant experiments under supervision and use this manual as an easy guide to learn experimental procedures. The content of the book minimizes the burden on the students and gives them the opportunity to have ample time in learning. Under the current curriculum the student is expected to maker relevant observations and record them too. It also contains depiction of important related images and figures which gives the student a comprehensive understanding of the subject and is useful in preparing for the viva voce examination. The practical manual will not only guide the students on relevant topics but their competency will also be simultaneously assessed by the teachers on daily basis.
Rumpa Saha MBBS, MD (Microbiology) is currently Professor, Department of Microbiology, University College of Medical Sciences and GTB Hospital, Delhi. She has been associated with this hospital since 2002. After completing her MBBS from Calcutta National Medical College, Calcutta University, she did her MD, Microbiology from PGIMER, Chandigarh. She has more than 16 years of teaching experience to PGs, UGs, nursing and MLT students. Her area of interest includes enteric diseases, parasitology and STIs. Besides conducting and supervising research in microbiology, she has to her credit 73 papers published in indexed national and international scientific journals of repute and has contributed chapters for Osteoarticular Tuberculosis, CBS Publishers & Distributors; Rational Use of Antimicrobials: Indian Perspectives, IMA Publications; Diploma in MLT Microbiology, IMA Publications, and has also edited the last two books. She has been examiner for MBBS, MD (Microbiology) and BDS of a number of universities. She is an active member of several scientific societies and has also delivered guest lectures and chaired sessions at a number of conferences / symposia in India.
Shukla Das MD, DNB, MNANS is currently Director-Professor, Department of Microbiology, University College of Medical Sciences and Guru Teg Bahadur Hospital, Delhi; former Assistant Director, National Centre for Disease Control, has to her credit more than 120 publications and 25 years of experience of teaching microbiology to medical undergraduates, postgraduates, nursing and dental students. She has guided several PhD students registered under Faculty Medical Sciences (FMS), Delhi University. Besides conducting teaching programs, she has completed 10 government funded projects in autoimmunity, diarrhoeal diseases and fungal infections. She has published 120 peer-reviewed and indexed papers and written chapters in 6 textbooks. She has been examiner for BDS, MBBS, MD and PhD examinations at several universities. She has been invited to deliver scientific talks at various national and international conferences. She is also the Joint Secretary, Indian Society for Human and Animal Mycology (SIHAM). She has been awarded several best paper poster/oral presentations of which the Satish Chandra best research paper award by IAMM, Delhi Chapter, in 2016, and Dr AM Saoji and Dr SR Sengupta memorial award at national conference MICROCON 2014 were the most coveted.
Science and technology are constantly changing fields. Newresearch and experience broaden the scope of information andknowledge. The authors have tried their best in giving informationavailable to them while preparing the material for this book.Although all efforts have been made to ensure optimum accuracyof the material, yet it is quite possible some errors might havebeen left uncorrected. The publisher, the printer and the authorswill not be held responsible for any inadvertent errors, orinaccuracies.
All rights reserved. No part of this book may be reproduced or transmitted in any form or by any means, electronic or mechanical,including photocopying, recording, or any information storage and retrieval system without permission, in writing, from the authorsand the publisher.
Published by Satish Kumar Jain and produced by Varun Jain for
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MicrobiologyPractical Manual
The existence of microorganism was predicted many centuries ago in Jain’s literature and by MarcusTerentous Varro in ancient Rome. Louis Pasteur and Robert Koch are the founders of medical
microbiology. The life expectancy has increased manyfolds over last 100 years after the development ofmicrobiological techniques. The understanding of microorganisms and microbiological techniques playsan important role for MBBS students in learning the biological basis of disease and its remedies. Thelearning process has to be gradual, stepwise and crystal clear.
It gives me great privilege to write the Foreword to the book Microbiology Practical Manual.The authors are well known to me for more than a decade and are very motivated and ambitious.
They are very experienced in teaching microbiology and are well versed in conducting undergraduateexaminations for MBBS students. I have observed keenness and steady craving in them to learn moreand endeavour towards perfection. Their eagerness to impart the sound education and create a clinicianwith clear understanding of the subject is being reflected in developing this manual.
This manual is structured to cover all the practical techniques of microbiology as is being taught andshown to them to achieve uniformity in learning as per the latest CBME guidelines. Hence, it would beof great benefit for microbiology students. I would recommend this manual for all MBBS students.
I congratulate the authors and am delighted to encourage and support them in pursuit of their objective.I wish the authors a grand success.
Dr Anil K JainMS FAMS FRCS (Eng.)
PrincipalDirector-Professor and HeadDepartment of Orthopaedics
University College of Medical Sciences and GTB HospitalDelhi
Foreword
I am indeed honored to write the Foreword to Microbiology Practical Manual.I really appreciate the efforts made by my colleagues Dr Shukla Das and Dr Rumpa Saha for
understanding the need of our students and in this endeavor enthusiastically preparing this practicalmanual as per the latest CBME guidelines.
They are our very dedicated teachers with a deep interest in the subject. As examiners, they are wellappreciated by the students.
This manual is very well planned and prepared with the objectives to envelop all the psychomotordomains of undergraduate microbiology and to educate the students to accomplish these tasks withease. It also aims to attain homogeneous learning. Thus, it would be of definite benefit to microbiologystudents. I would recommend this manual to all MBBS students.
I applaud the authors and am pleased to encourage them in quest of their objectives. I wish theauthors a grand success.
Dr NP SinghDirector-Professor and HeadDepartment of Microbiology
University College of Medical Sciences and GTB HospitalDelhi
Foreword
Microbiology Practical Manual 2nd edition has been prepared keeping in mind the latest guidelines of Competency Based Undergraduate Curriculum for the Indian Medical Graduate. It provides
the Phase II MBBS students relevant, comprehensive, reliable and informative approach to all practicalaspects of undergraduate microbiology as per CBME.
This manual contains all images of demonstration slides/specimens and other related items withcharacteristic features of each, relevant for undergraduate formative training.
For maintaining uniformity, all practical procedures have been clearly written and objectives aredefined to enable students to follow and recall.
We express our sincere thanks to our Principal, Director-Professor Dr A K Jain for his encouragementand for writing the Foreword for this manual. Our heartfelt thanks are also for our Head of the Department,Director-Professor Dr NP Singh for his constant support while preparing this manual and writing theForeword. Thanks are also due to M/s CBS Publishers & Distributors Pvt Ltd for their cooperation andkeen interest in the publication of this manual.
Rumpa SahaShukla Das
Preface
Section 1: General Microbiology Practicals
1. Rules and Safety Precautions to be Observed in the Microbiology Laboratory 32. GLP, Biohazard, Biosafety levels and Biosafety Cabinets 53. Universal Presence of Microbes 114. The Microscope: General Instruction about Use and Care 135. Microscopy and Micrometry 156. Methods of Sterilization and Disinfection 197. Biomedical Waste Disposal 238. Table of Commonly used Culture Media in Microbiology and their Uses 259. Demonstration of Common Culture Media and Biochemical Reactions 29
10. Culture Methods 4111. Description of the Appearance of Growth on Solid or in Liquid Media 4512. Demonstration of Methods of Antimicrobial Susceptibility Testing 4713. Miscellaneous Arthropods of Medical Importance (Including Tables) 5314. Demonstration of Methods for Diagnosis of Viral Infections 5915. Antigen–Antibody Reactions 63
Section 2: Perform and Identify the Different CausativeAgents of Infectious Diseases by Microscopy
16. Demonstration of Formol-ether Concentration Method for Stool Examination 7517. Demonstration of Normal Stool Findings 7718. Practical: Preparation of Saline and Iodine Wet Mount of Stool Sample for Diagnosis of Ova/Cysts 8119. Practical: To Stain Smear by Ziehl-Neelsen Technique for Diagnosis of Acid-fast Bacilli 8320. Practical: To Stain Smear by Albert Staining for Diagnosis of C. diphtheriae 8721. Practical: To Stain a Smear by Gram Stain for the Diagnosis of Bacterial Infections 8922. Practical: To Perform Motility Preparation of Bacteria for the Diagnosis of Bacterial Infections 9523. Demonstrations of other Staining Methods for Microscopy 97
i. Demonstration of Spore Staining and Anaerobic Methods for Diagnosis of Anaerobic Bacteria 97ii. Demonstration of Silver Impregnation Staining Method for Diagnosis of Spirochetes 103iii. Demonstration of LPCB Stain and Characteristic Features of the Common Fungi and their Diagnostic
Methods 105
Foreword by Dr Anil K Jain vForeword by Dr NP Singh viiPreface ix
Contents
xii Microbiology Practical Manual
Section 3: Identify the Causative Agent of Malaria and Filaria
24. Demonstration of Preparation of Blood Film 11125. Practical: To Study Peripheral Blood Smear Examination for the Characteristic Features for Malarial
Parasites 11726. Practical: To Study Peripheral Blood Smear Examination for the Characteristic Features for Microfilaria
for Diagnosis of Filariasis 12327. Demonstration of Morphological Forms of Leishmania donovani 125
Section 4: Identify the Common Etiologic Agents of Diarrhea and Dysentery
28. Table of Common Etiologic Agents Causing Diarrhea/Dysentery and their Method of Detection 13129. Practical: To Study the Characteristic Features of the Cysts of Entamoeba histolytica, Entamoeba coli
and Giardia lamblia 13330. Practical: To Study the Characteristic Features of the Eggs of Helminthic Cestodes 13731. Practical: To Study the Characteristic Features of the Eggs of Helminthic Nematodes 14132. Practical: To Study the Characteristic Features of the Oocysts of Coccidian Parasites 147
• Table of Common Ova/Cysts Found in Stool Microscopy 14833. Practical: To Study the Characteristic Features of Escherichia coli 15334. Practical: To Study the Characteristic Features of Shigella spp. 15535. Demonstration of the Characteristic Features of Vibrio cholerae 159
Section 5: Identify the Different Modalities for Diagnosis of Enteric Fever
36. Practical: To Study the Characteristic Features of Salmonella typhi 165
Section 6: Identify the Microbial Agents CausingRheumatic Heart Disease and Infective Endocarditis
37. Agents Causing Infective Endocarditis 17338. Practical: To Study the Characteristic features of Streptococcus pyogenes 175
Section 7: Identify the Common Etiological Agentsof Upper Respiratory Tract Infections (Gram Stain)
39. Table of Agents Causing Upper and Lower Respiratory Tract Infections 18140. Practical: To Study the Characteristic Features of C. diphtheriae 18541. Practical: To Study the Characteristic Features of Staphylococcus aureus 18742. Other Agents Causing Upper Respiratory Tract Infections, e.g. Staphylococcus aureus
(Revision Practical) 193
xiiiContents
Section 8: Identify the Common Etiological Agents of LowerRespiratory Tract Infections (Gram Stain and Acid-fast Stain)
43. To Study the Characteristic Features of Klebsiella spp. 19744. To Study the Characteristic Features of Mycobacterium tuberculosis 19945. To Study the Characteristic Features of Pseudomonas spp. 20346. Other Agents Causing Lower Respiratory Tract Infections, e.g. Staphylococcus aureus,
Section 9: Identify the Microbial Agents Causing Meningitis
47. Table of Agents Causing Meningitis 21348. Practical: To Study the Characteristic Features Gram-negative cocci Neissseria meningitidis 21949. Other agents causing meningitis, e.g. Staphylococcus aureus, Streptococcus spp, E. coli, Klebsiella spp.,
M. tuberculosis (Revision Practical) 221
Section 10: Demonstrate Infection Control Practices andUse of Personal Protective Equipment (PPE)
50. Practical: Demonstrate Hand Hygiene as an Infection Control Practice 22951. Practical: Demonstrate Types and Use of PPE 233
Section 11: Demonstrate the Appropriate Method of Collection of Samples in thePerformance of Laboratory Tests in the Detection of Microbial Agents
Causing Infectious Diseases
52. Specimens and Collection 239• Demonstrate Method of Collection of Blood Samples from Children and Adults 239• Demonstrate Method of Inoculation of Blood for Blood Culture in the Detection of Microbial Agents
Causing Septicemia 239• Amount, Number and Type of Blood Cultures 239• Transport and Incubation of Blood Culture Bottles 239• How to Minimize Contamination in Blood Culture 239• Other Types of Microbiological Specimens Used for Detection of Microorganisms Causing Infectious
Diseases 239• Criteria for Rejection of a Microbiological Sample 239• Transport and Storage of Microbiological Samples when Delay in Transport and Testing 239• Various Microbiology Specimens with Respect to Type of Infection 240
Section 12: Demonstrate Respect for Patient SamplesSent to the Laboratory for Performance of Laboratory Tests in the
Detection of Microbial Agents Causing Infectious Diseases
53. Practical: To be able to Demonstrate Need for Confidentiality Pertaining to Patient Identity with Regardsto Laboratory Results 255
1. Wash your hands with soap and water after handling the infectious material.
2. Wear laboratory coats in the laboratory.
3. Keep the nails clean and short.
4. Tie up long hair, while working in the laboratory.
5. Avoid eating, drinking and smoking in the laboratory.
6. Decontaminate the working area with appropriate disinfectant after spillage ofpotentially infected material.
7. Carry-out the laboratory procedure following standard precautions.
8. Avoid mouth pipetting as far as possible.
9. Perform all the laboratory procedures in a way that minimizes the aerosol formation.
10. Bring practical manual in all the practical classes.
11. Draw properly labeled diagrams neatly, get it corrected from your teachers.
12. Leave your microscope and work seat clean before going out of practical lab.
13. Keep your bag away from working tables.
14. Come exactly at 2.00 pm for practical and tutorials.
Rules and Safety Precautions to beObserved in the Microbiology Laboratory
