DOCSLIB.ORG

Basic Practical Microbiology a MANUAL

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Basic Practical Microbiology a MANUAL 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
Load more

Recommended publications
  • 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 ................................................................................
    [Show full text]
  • Exploring Small-Scale Chemostats to Scale up Microbial Processes: 3-Hydroxypropionic Acid Production in S
    Lawrence Berkeley National Laboratory Recent Work Title Exploring small-scale chemostats to scale up microbial processes: 3-hydroxypropionic acid production in S. cerevisiae. Permalink https://escholarship.org/uc/item/43h8866j Journal Microbial cell factories, 18(1) ISSN 1475-2859 Authors Lis, Alicia V Schneider, Konstantin Weber, Jost et al. Publication Date 2019-03-11 DOI 10.1186/s12934-019-1101-5 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Lis et al. Microb Cell Fact (2019) 18:50 https://doi.org/10.1186/s12934-019-1101-5 Microbial Cell Factories RESEARCH Open Access Exploring small-scale chemostats to scale up microbial processes: 3-hydroxypropionic acid production in S. cerevisiae Alicia V. Lis1, Konstantin Schneider1,2, Jost Weber1,3, Jay D. Keasling1,4,5,6,7, Michael Krogh Jensen1 and Tobias Klein1,2* Abstract Background: The physiological characterization of microorganisms provides valuable information for bioprocess development. Chemostat cultivations are a powerful tool for this purpose, as they allow defned changes to one single parameter at a time, which is most commonly the growth rate. The subsequent establishment of a steady state then permits constant variables enabling the acquisition of reproducible data sets for comparing microbial perfor- mance under diferent conditions. We performed physiological characterizations of a 3-hydroxypropionic acid (3-HP) producing Saccharomyces cerevisiae strain in a miniaturized and parallelized chemostat cultivation system. The physi- ological conditions under investigation were various growth rates controlled by diferent nutrient limitations (C, N, P). Based on the cultivation parameters obtained subsequent fed-batch cultivations were designed. Results: We report technical advancements of a small-scale chemostat cultivation system and its applicability for reliable strain screening under diferent physiological conditions, i.e.
    [Show full text]
  • Microscope Compatible Cell-Culture Incubator
    Microscope Compatible Cell-Culture Incubator BME 400 December 14th, 2016 Client: Dr. John Puccinelli, UW-Madison Dept. of Biomedical Engineering Advisor: Dr. Mitchell Tyler, UW-Madison Dept. of Biomedical Engineering Team Members: Trevor Zarecki - Team Leader Jenny Westlund - Communicator Peter Hartig - BPAG Jack McGinnity - BWIG Steve Gock - BSAC Abstract Live cell imaging experiments are difficult to perform over long periods of time on normal lab microscopes without incubation capabilities. Current commercial microscope-stage incubators are expensive, not compatible with multiple microscopes and ineffective at evenly controlling the environment. The client desires an inexpensive incubation chamber compatible with cell microscopy that is capable of maintaining desired temperature, CO2, and humidity evenly throughout the chamber. The device should not alter image quality, and should be accessible for changing media or cell culture dishes. An initial prototype has been developed that involves a small, cohesive system to regulate temperature, CO2, and humidity. Testing of the current prototype has demonstrated regulation of these three systems, through an automated feedback system capable of maintaining the system near a physiological set point. Further development of the design will ensure that it performs efficiently according to all specifications, and ultimately help bridge the gap in the market between high-cost, functional incubation systems and cheaper, less effective designs. 1 Table of Contents Introduction 3 Motivation 3 Existing
    [Show full text]
  • Use of Laboratory Equipement
    USE OF LABORATORY EQUIPEMENT C. Laboratory Thermometers Most thermometers are based upon the principle that liquids expand when heated. Most common thermometers use mercury or colored alcohol as the liquid. These thermometers are constructed as that a uniform-diameter capillary tube surmounts a liquid reservoir. To calibrate a thermometer, one defines two reference points, normally the freezing point of water (0°C, 32°F) and the boiling point of water (100°C, 212°F) at 1 tam of pressure (1 tam = 760 mm Hg). Once these points are marked on the capillary, its length is then subdivided into uniform divisions called degrees. There are 100° between these two points on the Celsius (°C, or centigrade) scale and 180° between those two points on the Fahrenheit (°F) scale. °F = 1.8 °C + 32 The Thermometer and Its Calibration This section describes the proper technique for checking the accuracy of your thermometer. These measurements will show how measured temperatures (read from thermometer) compare with true temperatures (the boiling and freezing points of water). The freezing point of water is 0°C; the boiling point depends upon atmospheric pressure but at sea level it is 100°C. Option 1: Place approximately 50 mL of ice in a 250-mL beaker and cover the ice with distilled water. Allow about 15 min for the mixture to come to equilibrium and then measure and record the temperature of the mixture. Theoretically, this temperature is 0°C. Option 2: Set up a 250-mL beaker on a wire gauze and iron ring. Fill the beaker about half full with distilled water.
    [Show full text]
  • Laboratory Glassware N Edition No
    Laboratory Glassware n Edition No. 2 n Index Introduction 3 Ground joint glassware 13 Volumetric glassware 53 General laboratory glassware 65 Alphabetical index 76 Índice alfabético 77 Index Reference index 78 [email protected] Scharlau has been in the scientific glassware business for over 15 years Until now Scharlab S.L. had limited its sales to the Spanish market. However, now, coinciding with the inauguration of the new workshop next to our warehouse in Sentmenat, we are ready to export our scientific glassware to other countries. Standard and made to order Products for which there is regular demand are produced in larger Scharlau glassware quantities and then stocked for almost immediate supply. Other products are either manufactured directly from glass tubing or are constructed from a number of semi-finished products. Quality Even today, scientific glassblowing remains a highly skilled hand craft and the quality of glassware depends on the skill of each blower. Careful selection of the raw glass ensures that our final products are free from imperfections such as air lines, scratches and stones. You will be able to judge for yourself the workmanship of our glassware products. Safety All our glassware is annealed and made stress free to avoid breakage. Fax: +34 93 715 67 25 Scharlab The Lab Sourcing Group 3 www.scharlab.com Glassware Scharlau glassware is made from borosilicate glass that meets the specifications of the following standards: BS ISO 3585, DIN 12217 Type 3.3 Borosilicate glass ASTM E-438 Type 1 Class A Borosilicate glass US Pharmacopoeia Type 1 Borosilicate glass European Pharmacopoeia Type 1 Glass The typical chemical composition of our borosilicate glass is as follows: O Si 2 81% B2O3 13% Na2O 4% Al2O3 2% Glass is an inorganic substance that on cooling becomes rigid without crystallising and therefore it has no melting point as such.
    [Show full text]
  • Detection of Acid-Producing Bacteria Nachweis Von Säureproduzierenden Bakterien Détection De Bactéries Produisant Des Acides
    (19) TZZ ¥ _T (11) EP 2 443 249 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12Q 1/04 (2006.01) G01N 33/84 (2006.01) 19.11.2014 Bulletin 2014/47 (86) International application number: (21) Application number: 10790013.6 PCT/US2010/038569 (22) Date of filing: 15.06.2010 (87) International publication number: WO 2010/147918 (23.12.2010 Gazette 2010/51) (54) DETECTION OF ACID-PRODUCING BACTERIA NACHWEIS VON SÄUREPRODUZIERENDEN BAKTERIEN DÉTECTION DE BACTÉRIES PRODUISANT DES ACIDES (84) Designated Contracting States: (74) Representative: Isarpatent AL AT BE BG CH CY CZ DE DK EE ES FI FR GB Patent- und Rechtsanwälte GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO Friedrichstrasse 31 PL PT RO SE SI SK SM TR 80801 München (DE) (30) Priority: 15.06.2009 US 187107 P (56) References cited: 15.03.2010 US 314140 P US-A- 4 528 269 US-A- 5 098 832 US-A- 5 164 301 US-A- 5 601 998 (43) Date of publication of application: US-A- 5 601 998 US-A- 5 786 167 25.04.2012 Bulletin 2012/17 US-B2- 6 756 225 US-B2- 7 150 977 (73) Proprietor: 3M Innovative Properties Company • DARUKARADHYA J ET AL: "Selective Saint Paul, MN 55133-3427 (US) enumeration of Lactobacillus acidophilus, Bifidobacterium spp., starter lactic acid bacteria (72) Inventors: and non-starter lactic acid bacteria from Cheddar • YOUNG, Robert, F. cheese", INTERNATIONAL DAIRY JOURNAL, Saint Paul, Minnesota 55133-3427 (US) ELSEVIER APPLIED SCIENCE, BARKING, GB, • MACH, Patrick, A.
    [Show full text]
  • Chemostat Culture for Yeast Experimental Evolution
    Downloaded from http://cshprotocols.cshlp.org/ at Cold Spring Harbor Laboratory Library on August 9, 2017 - Published by Cold Spring Harbor Laboratory Press Protocol Chemostat Culture for Yeast Experimental Evolution Celia Payen and Maitreya J. Dunham1 Department of Genome Sciences, University of Washington, Seattle, Washington 98195 Experimental evolution is one approach used to address a broad range of questions related to evolution and adaptation to strong selection pressures. Experimental evolution of diverse microbial and viral systems has routinely been used to study new traits and behaviors and also to dissect mechanisms of rapid evolution. This protocol describes the practical aspects of experimental evolution with yeast grown in chemostats, including the setup of the experiment and sampling methods as well as best laboratory and record-keeping practices. MATERIALS It is essential that you consult the appropriate Material Safety Data Sheets and your institution’s Environmental Health and Safety Office for proper handling of equipment and hazardous material used in this protocol. Reagents Defined minimal medium appropriate for the experiment For examples, see Protocol: Assembly of a Mini-Chemostat Array (Miller et al. 2015). Ethanol (95%) Glycerol (20% and 50%; sterile) Yeast strain of interest Equipment Agar plates (appropriate for chosen strain) Chemostat array Assemble the apparatus as described in Miller et al. (2013) and Protocol: Assembly of a Mini-Chemostat Array (Miller et al. 2015). Cryo deep-freeze labels Cryogenic vials Culture tubes Cytometer (BD Accuri C6) Glass beads, 4 mm (sterile; for plating yeast cells) Glass cylinder Kimwipes 1Correspondence: [email protected] © 2017 Cold Spring Harbor Laboratory Press Cite this protocol as Cold Spring Harb Protoc; doi:10.1101/pdb.prot089011 559 Downloaded from http://cshprotocols.cshlp.org/ at Cold Spring Harbor Laboratory Library on August 9, 2017 - Published by Cold Spring Harbor Laboratory Press C.
    [Show full text]
  • Annual Conference Abstracts
    ANNUAL CONFERENCE 14-17 April 2014 Arena and Convention Centre, Liverpool ABSTRACTS SGM ANNUAL CONFERENCE APRIL 2014 ABSTRACTS (LI00Mo1210) – SGM Prize Medal Lecture (LI00Tu1210) – Marjory Stephenson Climate Change, Oceans, and Infectious Disease Prize Lecture Dr. Rita R. Colwell Understanding the basis of antibiotic resistance University of Maryland, College Park, MD, USA as a platform for early drug discovery During the mid-1980s, satellite sensors were developed to monitor Laura JV Piddock land and oceans for purposes of understanding climate, weather, School of Immunity & Infection and Institute of Microbiology and and vegetation distribution and seasonal variations. Subsequently Infection, University of Birmingham, UK inter-relationships of the environment and infectious diseases Antibiotic resistant bacteria are one of the greatest threats to human were investigated, both qualitatively and quantitatively, with health. Resistance can be mediated by numerous mechanisms documentation of the seasonality of diseases, notably malaria including mutations conferring changes to the genes encoding the and cholera by epidemiologists. The new research revealed a very target proteins as well as RND efflux pumps, which confer innate close interaction of the environment and many other infectious multi-drug resistance (MDR) to bacteria. The production of efflux diseases. With satellite sensors, these relationships were pumps can be increased, usually due to mutations in regulatory quantified and comparatively analyzed. More recent studies of genes, and this confers resistance to antibiotics that are often used epidemic diseases have provided models, both retrospective and to treat infections by Gram negative bacteria. RND MDR efflux prospective, for understanding and predicting disease epidemics, systems not only confer antibiotic resistance, but altered expression notably vector borne diseases.
    [Show full text]
  • Thank You for Purchasing This Tesla Immersion Heater. This Unit Is
    Specialist Immersion Heaters by Tesla UK Thank you for purchasing this Tesla Immersion Heater. This unit is guaranteed for a period of 12 months from the date of receipted purchase provided that it has been installed correctly by a suitably qualified person. The installer must ensure: • The immersion heater is installed in a system where the heating element is always below water level. • There is water in the tank before the immersion heater is first switched on. • It is only subjected to normal operating conditions in a domestic hot water system which conforms to BS699, BS1566 or BS3198 and in which the system temperature is no more than 75°C. No warranty is hereby given or implied in other uses except domestic. • It must be fitted in accordance with current IEE wiring regulations and must be wired through a double pole isolator or suitable controller which must have a contact separation of at least 3mm in all poles. This appliance can be used by children aged from 8 years and above and persons with reduced physical, sensory or mental capabilities or lack of experience and knowledge if they have been given supervision or instruction concerning us of the appliance in a safe way and understand the hazards involved. Children shall not play with the appliance. Cleaning and user maintenance shall not be made by children without supervision. This unit must not be modified in any way. BEAB approval for all models is dependent upon the fitting of the appropriate Dual Safety Thermostat that is listed in these instructions. Tesla UK Ltd, Unit 3b First Avenue, Minworth, Sutton Coldfield, B76 1BA Tel: +44 (0) 121 686 8711 Technical: +44 (0) 121 686 8733 [email protected] www.teslauk.com Specialist Immersion Heaters by Tesla UK General Fitting Information The Aquatherm range of immersion heaters are direct equivalents to the immersion heaters fitted to Heatrae Sadia Megaflo cylinders.
    [Show full text]
  • Autoclave Quick Guide
    Office of Biological Safety Autoclave Operation Quick Reference Guide Training: ALL users MUST undergo documented training for operation of the autoclave. Record each use of the autoclave in a log: Date, User ID, cycle type and nature of material in load Personal Protective Equipment (PPE) needed: ¾ Wear eye protection, lab coat, gloves along with heat resistant gloves. Rubberized apron, sleeve guards and face shield are recommended when autoclave is hot or splash risk is present. Hints and Precautions ¾ Become familiar with the manufacturer’s operations manual of your autoclave model(s). ¾ Plastics used for autoclaving MUST be labeled as autoclavable – otherwise the plastic will melt. ¾ Waste bags to be autoclaved must be loosely packed and not more than 2/3 filled. Steam must be able to penetrate to all contents of the bag. ¾ Sharps or pointed hard objects should not be placed directly into an autoclave bag; a thicker or rigid container must be used (such as a sharps container). ¾ Avoid overfilling an autoclave with loads or allowing a load to contact the chamber walls. ¾ Transferring waste contents from an overfilled bag to another bag should be avoided! This practice can lead to injury and/or exposure to contaminants. ¾ Do not leave an autoclave operating unattended for long periods of time; operation should be monitored periodically during a cycle in case of failure. ¾ Never autoclave solvents, combustible, volatile, flammable, radioactive or corrosive materials (e.g. ethanol, methanol, acids, bases, phenol) ¾ Remove extraneous items and combustible materials from around the autoclave exterior. Loading ¾ Follow manufacturer’s loading instructions for your autoclave model ¾ Transport loads on a cart and in secondary containers to reduce spills ¾ Clean item/container sterilization Loosen caps or lids to avoid dangerous pressure build-up during cycle Place containers in a tray and load the tray into the autoclave – this is easier to load and unload and will catch spills.
    [Show full text]
  • Performance Validation of the Microbiologique Microfilm Test
    MAI ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 101, NO. X, 2018 1 FOOD BIOLOGICAL CONTAMINANTS Performance Validation of the Microbiologique MicrofilmTM Test System for AOAC Research Institute Performance Tested Method SMCertification AOAC Performance Tested Method SM 051702 Abstract ANNA SHAPOVALOVA and HARISH K. JANAGAMA Molecular Epidemiology, Inc., 15300 Bothell Way NE, The Microfilm™ Test System is intended for quantitative Lake Forest Park, WA 98155 microbiology and consists of three types of Microfilms for aerobic LONG VUONG and ALEX FRIEDRICH plate count (Microfilm APC), total coliform and Escherichia coli IEH Laboratories and Consulting Group, Inc., 15300 Bothell count (Microfilm TCEc), and yeast and mold count (Microfilm Way NE, Lake Forest Park, WA 98155 YMC). This study evaluated the performance of the Microfilm DYLAN JOHNSON Test System against International Organization for Standardization Molecular Epidemiology, Inc., 15300 Bothell Way NE, (ISO) methods on 20 food matrixes and 2 environmental surfaces. Lake Forest Park, WA 98155 Ruggedness, robustness, and stability were also determined, LOURDES M. NADALA while inclusivity and exclusivity studies were performed on Molecular Epidemiology, Inc., 15300 Bothell Way N.E., Microfilm TCEc and YMC. An independent laboratory evaluated Lake Forest Park, WA 98155; Microbiologique, the performance on four food matrixes and one environmental Inc., 8215 Lake City Way NE, Seattle, WA 98115 surface. No significant differences and high correlation coefficients VAN NGUYEN were observed between the Microfilm Test System and the Microbiologique, Inc., 8215 Lake City Way NE, Seattle, corresponding ISO methods (ISO 4833-1:2013 for APC, ISO WA 98115 4832:2006 for total coliform count, ISO 16649-2: 2001 for E.
    [Show full text]
  • 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.
    [Show full text]