Basic Practical Microbiology A MANUAL Microbiology Society Basic Pract Cover 2009 16/1/09 12:01 Page 4 About this resource Microbiology is a popular option for practical work in schools. This manual, which explains the basic Abouttechniques necessarythis resource to carry out microbiology experiments safely and effectively, is intended as a guide for teachers and technicians. It forms the notes for the one-day basic practical microbiology Microbiologytraining course is whicha popular is run option and foraccredited practical by work the Societyin schools. for ThisGeneral manual, Microbiology which explains (SGM), the but basic can techniquesbe used as a necessary standalone to document. carry out microbiologyFurther information experiments about safelythe training and effectively, courses is isavailable intended on as the a guideSGM’s foreducation teachers website and technicians. (see below), which also includes a wealth of other material relating to both Thepractical Microbiology and theoretical Society, microbiologyin association teaching. with the Microbiology in Schools Advisory Committee (MiSAC) hasThe alsoSGM, produced in association a book withof 21 the practical Microbiology investigations in Schools which Advisory complements Committee the manual (MISAC) – Practicalhas also Microbiologyproduced a forbook Secondary of 21 practicalSchools isinvestigations suitable for use which with complementsKey Stages 3, 4the and manual: post-16 Practical and the equivalentMicrobiology Scottish for Secondary qualifications. Schools Thisis suitableresource for can use be withdownloaded Key Stages from 3, the 4 andMicrobiology post-16 andSociety’s the dedicatedequivalent education Scottish qualifications. website: www.microbiologyonline.org.uk Ordering details are available on the website noted below. The MicrobiologySGM offers schoolsSociety membershipoffers schools and membership runs an advice and runs service an adviceon microbiology service on teaching.microbiology For teaching.further details, For further email details,education [email protected], [email protected], telephone +44 (0)118 988 telephone1835, fax +44 (0)20(0)118 7685 988 26825656 oror writewrite to to the the address address at at the the bottom bottom of theof thepage. page. Much information can be found on the Otherwebsite: useful www.microbiologyonline.org.uk sources of advice: xOther CLEAPSS useful –sources www.cleapss.org.uk of advice: ᭟ The Gardiner www.cleapss.org.uk Building, Brunel Science Park, Uxbridge UB8 3PQ x BrunelMicrobiology University, in Schools Uxbridge Advisory UB8 3PH Committee (MiSAC) – www.misac.org.uk ᭟ icrobiologyc/o NCBE (see inaddress cool below) iory ommiee www.microbiologyonline.org.uk/misac x MarlboroughNational Centre House, for Biotechnology Basingstoke Road, Education Spencers (NCBE) Wood, – www.ncbe.reading.ac.uk Reading RG7 1AG ᭟ ionlScience and enre Technology or ioecnology Centre, The ucion University of Reading, www.ncbe.reading.ac.uk 2 Earley Gate, Whiteknights, ScienceReading and RG6 Technology 6AU Centre, The University of Reading, Earley Gate, PO Box 247, x ReadingScottish RG6Schools 6BZ Equipment Research Centre (SSERC) – www.sserc.org.uk ᭟ coi2 Pitreavie cool Court, uimenSouth Pitreavie eerc Business enre Park, Dunfermline www.sserc.org.uk KY11 8UB 2 Pitreavie Court, South Pitreavie Business Park, Dunfermline KY11 8UB Credits && AcknowledgementsAcknowledgements The MicrobiologySGM gratefully Society acknowledges gratefully the acknowledges support from the the support following from sources: the following sources: ᭟x Kath Crawford (SAPS Scotland) ᭟x John Richardson (SSERC) ᭟x Members of MiSACMISAC ᭟x John Schollar (NCBE) ᭟x John Tranter (CLEAPSS) Editors: Dariel Burdass, John Grainger & Janet Hurst Design and Production Editor: Ian Atherton Photographs: Dariel Burdass & Faye Jones Front cover illustration: TEKWladimir Image Bulgar/Science / Science Photo Photo Library Library ISBN 0 95368 383 4 Copyright Basic Practical Microbiology – A Manual is copyright. The Society for General Microbiology asserts itsBasic moral Practical right toMicrobiology be identified – asA Manual copyright is holdercopyright. under The Section Microbiology 77 of the Society Designs, asserts Patents its moraland Copyrightright to be Act identified (UK) 1988. as copyright holder under Section 77 of the Designs, Patents and Copyright Act (UK) 1988. Educational use: Electronic or paper copies of the resource or individual pages from it may beEducational made for classroomuse: Electronic and bona or paper fide educationalcopies of the uses, resource provided or individualthat the copies pages are from distributed it may be freemade of for charge classroom or at and the bona cost fideof reproduction educational anduses, thatprovided the SGMthat theis creditedcopies are and distributed identified free as of copyrightcharge or holder.at the cost of reproduction and that the Microbiology Society is credited and identified as copyright holder. Published by the Society for General Microbiology, Marlborough House, Basingstoke Road, SpencersPublished Wood, by the Reading Microbiology RG7 1AG, Society, UK Charles Darwin House, 12 Roger Street, London WC1N 2JU, UK © 2006 Society for General Microbiology © 2016 Microbiology Society Basic Pract Book 2006 2/11/06 11:17 am Page 1 Contents About this resource Inside front cover Part 1: The Basics An introduction to microbiology, aseptic technique and safety Preparation Safety guidelines 1 Risk assessment 2 Good microbiological laboratory practice (GMLP) 3 Spillage management 3 Aerosols 3 Resources Equipment 4 Apparatus 5 Materials 5 Media, sterilisation and disinfection Preparation of culture media 6 Pouring a plate 6 Storage of media 6 Sterilisation vs disinfection 6 Sterilisation using the autoclave/pressure cooker 7 Sterilisation of equipment and materials 7 Choice, preparation and use of disinfectants 7 Inoculation and other aseptic procedures Essential points 8 Using a wire loop 8 Using a pipette 9 Flaming the neck of bottles and test tubes 10 Working with bacteria and yeast Streak plate 11 Pour plate 12 Using a spreader 13 Spread plate 14 Working with moulds 15 Incubation 16 In conclusion: clearing up 17 Basic Pract Book 2006 2/11/06 11:17 am Page 2 Essential methods for maintaining, preparing and using cultures Obtaining suitable cultures 18 Pure cultures 18 Maintaining stock cultures 18 Checking cultures for contamination 18 Preventing contamination of cultures and the environment 19 Aseptic transfer of cultures and sterile solutions 19 Preparing cultures for class use 19 Part 2: Microbiology in Action Practical activities 1. Testing sensitivity to antimicrobial substances 21 2. Microscopy Using the microscope 22 Stained preparations 23 Making a smear 23 A simple stain 24 A differential stain: Gram’s staining method 24 Appendices 1. Safety guidelines 26 2. Safe micro-organisms 31 3. Safety resources 37 4. Suppliers of cultures and equipment 38 5. Use of the autoclave/pressure cooker 39 6. Preparing serial dilutions 40 Basic Pract Book 2006 2/11/06 11:17 am Page 1 Basic Pract PartBook 2006 1: 2/11/06 The 11:17 Basics am Page 1 An introduction to microbiology, aseptic technique and safety As well as causing a familiar range of diseases in animals and plants and problems in food spoilage and deterioration Partof other materials,1: The microbes Basics are also our ‘invisible allies’. Indeed, life on Earth would not be sustainable without the Anbenefits introduction that many of them to provide. microbiology, The teaching of suchaseptic an important subject as microbiology cannot be achieved effectively without enhancing the theory with ‘hands on’ experience in the laboratory. The purpose of this manual is to techniqueprovide teachers andand technicians safety with good techniques in practical microbiology to ensure that investigations proceed safely and achieve the required educational aims successfully. This manual has been written for a right-handed person. As well as causing a familiar range of diseases in animals and plants and problems in food spoilage and deterioration of other materials, microbes are also our ‘invisible allies’. Indeed, life on Earth would not be sustainable without the benefits that many of them provide. The teaching of such an important subject as microbiology cannot be achieved effectively without enhancing the theory with ‘hands on’ experience in the laboratory. The purpose of this manual is to Preparationprovide teachers and technicians with good techniques in practical microbiology to ensure that investigations proceed safely and achieve the required educational aims successfully. This manual has been written for a right-handed person. PreparationSafety guidelines The small size of microbes and the consequent need to deal with cultures that contain many millions of microbial cells require special procedures for their safe use. Activities involving micro-organisms are controlled by thethe ControlControl ofof Substances Hazardous to Health (COSHH) Regulations and teachers and technicians have a duty under the Health and Safety at Work Act to comply with any safety instructions given by their employers. These include using model risk assessments.assessments forThe which publications it is necessary normally to referused toin appropriatesecondary schools publications are: such as CLEAPSS Laboratory Handbook (2006), section 15.2, Topics in Safety, 3rd edition (ASE, 2001), Microbiology: an HMI Guide (DES, 1990) and Safety in Science xSafety Topics guidelines