1、third edition Basic Microbiology for Drinking Water Dennis R. Hill Basic Microbiology-AWWA.indb 1 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.Basic Microbiology for Drinking Water, Third Edition Copyright 2014 American Water Works Association All rights reser
2、ved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information or retrieval system, except in the form of brief excerpts or quotations for review purposes, without the written permission of t
3、he publisher. Disclaimer The authors, contributors, editors, and publisher do not assume responsibility for the valid- ity of the content or any consequences of its use. In no event will AWWA be liable for direct, indirect, special, incidental, or consequential damages arising out of the use of info
4、rmation presented in this book. In particular, AWWA will not be responsible for any costs, includ- ing, but not limited to, those incurred as a result of lost revenue. In no event shall AWWAs liability exceed the amount paid for the purchase of this book. The use of trade names or discussion of pate
5、nted products and processes in this text does not in any way constitute an endorsement or otherwise by the author, publisher, or editor.AWWA Senior Manager of Editorial Development and Production: Gay Porter De Nileon AWWA Senior Technical Editor/Project Manager: Martha Ripley Gray Cover Art: Melani
6、e Yamamoto Production: Janice Benight Design Studio Original line drawings, photographs, and other artwork in this volume have been composed by Dennis R. Hill, in his capacity as microbiologist for the Des Moines Water Works. Library of Congress Cataloging-in-Publication Data Hill, Dennis, 1951-Basi
7、c microbiology for drinking water / Dennis R. Hill. - Third edition.pages cmIncludes bibliographical references and index.ISBN 978-1-58321-981-2 - ISBN 978-1-61300-258-2 (electronic) 1. Drinking water-Microbi- ology. 2. Drinking water-Purification. I. American Water Works Association. II. Title. TD3
8、84.H56 2014579.0246281-dc23 ISBN 978-1-58321-981-2; eISBN 978-1-61300-258-2 Printed in the United States of America American Water Works Association 6666 West Quincy Avenue Denver, CO 80235-3098 awwa.org Printed on Recycled Paper Basic Microbiology-AWWA.indb 2 11/4/13 4:15 PM Copyright 2014 American
9、 Water Works Association. All Rights Reserved.Basic Microbiology 3 rded. iii Figures, vii Tables, xi Preface, xiii Acknowledgments, xiii 1 Introduction 1 Drinking Water Microbiology, 1 Disease Transmission, 2 Immunology, 5 Microbial Environmental Diversity, 6 Organization of the Book, 8 2 Bacteria 9
10、 Bacterial Metabolism, 9 Bacterial Toxins, 10 Bacteria of Interest to the Water Industry, 12 Gram-Negative, 12 Escherichia coli, 12 Shigella, 13 Salmonella, 13 Aeromonas and Plesiomonas, 15 Vibrio cholerae, 15 Campylobacter, 15 Helicobacter pylori, 16 Yersinia entercolitica, 16 Legionella pneumophil
11、a, 17 Pseudomonas aeruginosa, 17 Mycobacterium avium-intracellulare, 18 Gram-Positive, 19 Staphylococcus Species, 19 Streptococcus Species, 20 Enterococcus faecalis, 20 Clostridium perfringens, 20 Bacillus Species, 21 B. cereus and Other Bacillus Species, 22 Contents Basic Microbiology-AWWA.indb 3 1
12、1/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.iv Basic Micro Biol ogy for drinking water Basic Microbiology 3 rded. Nuisance Bacteria, 22 Actinomycetes, 22 Iron Bacteria, 22 Sulfur Bacteria, 23 Nitrifying Bacteria, 24 Denitrifying Bacteria, 24 Cyanobacteria (Blu
13、e-Green Algae), 26 3 Viruses, Protists, and Other Organisms .31 Viruses, 31 Hepatitis A Virus, 32 Enteroviruses (Poliovirus, Coxsackievirus, Echovirus, and Others), 32 Gastroenteritis Viruses, 33 Protists, 33 Protozoan-like Protists, 33 Paramecium, 34 Giardia lamblia, 35 Cryptosporidium parvum, 36 C
14、yclospora cayetanensis, 37 Microsporidia Group, 37 Amoebae, 38 Algal-Like Protists, 40 Multicellular Organisms, 46 4 Laboratory Methods for Pathogen Isolation and Detection .49 Sampling, 50 Building a Progressive Microbiology Laboratory, 50 Progressive Microbiology Laboratory Battery, 51 USEPA Total
15、 Coliform Rule, 52 Conventional Testing Methods, 53 Heterotrophic Plate Count, 56 MMO-MUG Media, 57 Membrane Filtration, 58 Media Summary, 61 Proper Incubation Time, 65 Specific Coliform Detection Media, 67 Membrane Filter Integrity Study, 68 Basic Microbiology-AWWA.indb 4 11/4/13 4:15 PM Copyright
16、2014 American Water Works Association. All Rights Reserved.contents v Basic Microbiology 3 rded. Cost Study: Membrane Filtration and MMO-MUG Media, 68 Testing for Protozoa, 70 Testing for Viruses, 72 Testing for Coliphages, 73 Methods of Testing for Other Organisms, 74 Advances in Molecular Biology,
17、 74 Light Microscopy, 74 Kohler Illumination, 77 Aerobic Endospore Method, 77 Well Slide and Glycerol Phytoplankton Method, 82 Chlorine Contact Time Value, 87 Effective Data Communication, 87 Measurements and Data Accuracy, 89 5 Chemistry of Microbiology and Water Treatment .91 Inorganic Chemistry,
18、92 Periodic Table of the Elements, 92 Chemical Compounds, 94 Chemical Formulas and Notation, 95 Acids and Bases, 96 Organic Chemistry, 97 The Nitrogen Cycle, 98 Microbiology in Water Treatment, 100 Softening, 101 Disinfection, 102 Appendix A Scientific Nomenclature, Notation, and Numerical Prefixes
19、and Abbreviations, 105 Appendix B Preparation of New Water Mains for Service, 107 Bibliography, 113 Additional Resources, 115 Index, 117 About the Author, 125 Basic Microbiology-AWWA.indb 5 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.Basic Microbiology-AWWA.i
20、ndb 6 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.Basic Microbiology 3 rded. vii 1-1 Microorganisms size comparison, 8 2-1 Gram-negative rods, 11 2-2 Gram-positive cocci, 11 2-3 Mycobacterial stain, 19 2-4 Streptococcus colonies on sheep blood agar plate, 20
21、2-5 Beggiatoa, 23 2-6 Pipe tubercle, 23 2-7 Pond used for nitrate removal, 25 2-8 Experimental ponds showing cyanobacterial growth when stagnant and minimal growth when mixed, 25 2-9 Cylindrospermopsis, 27 2-10 Microcystis, 27 2-11 Oscillatoria, 28 2-12 Anabaena with reproductive cells, 28 2-13 Plan
22、ktothrix, 29 2-14 Aphanizomenon, 29 2-15 Dactylococcopsis cyanobacterium, 30 3-1 Paramecia, 34 3-2 Giardia cysts, 35 3-3 Cryptosporidium oocysts, 37 3-4 Amoeba on filamentous algae strands, 39 3-5 Unicellular alga, 41 3-6 Unicellular algae, 41 3-7 Dinoflagellate, 41 3-8 Pennate diatom, 43 3-9 Coloni
23、al algae Pediastrum, 43 3-10 Colonial algae Astrionella, 43 3-11 Cyclotella diatomaceous alga, 44 3-12 Filamentous algae, 44 3-13 Filamentous alga Spirogyra, 45 Figures Basic Microbiology-AWWA.indb 7 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.viii Basic Micr
24、o Biol ogy for drinking water Basic Microbiology 3 rded. 3-14 Planktonic algae, 45 3-15 Euglena, 46 3-16 Nematode worm, 47 3-17 Rotifer, 48 3-18 Water flea with eggs, 48 4-1 Microbiology laboratory, 51 4-2 Agar plate with colonies: MacConkey agar with pink colonies that express lactose fermentation,
25、 54 4-3 Agar plate with colonies: MacConkey agar with clear colonies that express no lactose fermentation, 55 4-4 API 20E biochemical strip, 56 4-5 Broth tubes with growth: (A) Lactose tryptose broth; (B) Brilliant green lactose bile broth; and (C) EC+MUG broth with UV light, 56 4-6 HPC plate with c
26、olonies, 57 4-7 Colilert TMmedium packet, 59 4-8 Colilert TMbottles: (A) No growth; (B) Growth of total coliforms; and (C) Growth of E. coli (with UV light), 59 4-9 Water bath with presence/absence Colilert TMvessels, 59 4-10 Quanti-tray 2000 with no growth, 60 4-11 Quanti-tray 2000 with total colif
27、orm bacteria, 60 4-12 Quanti-tray 2000 with E. coli (fluorescence upon UV light exposure), 60 4-13 mENDO medium for membrane filtration, 63 4-14 mENDO plate with sheen colonies, 63 4-15 Vacuum manifold for membrane filtration, 64 4-16 Bacterial growth curve over 24 hours and Colilert TMmedium color
28、(yellow shading indicates color of the medium), 66 4-17 Colilert TMmedium at 20, 22, and 24 hours, 66 4-18 Media and method cost comparison, 70 4-19 Coliphage-MS2 plaques on a lawn of E. coli, 73 4-20 Leeuwenhoek microscope, 75 4-21 Modern microscope, 76 4-22 Membrane filtration apparatus, 79 4-23 C
29、ookie sheet with filters, 79 Basic Microbiology-AWWA.indb 8 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.figures ix Basic Microbiology 3 rded. 4-24 Colonies derived from endospores, 80 4-25 Endospore counting cover, 81 4-26 Well slide with samples, 83 4-27 Cen
30、trifuge tube, 84 4-28 Collage of microbes on well slide, 200, 86 4-29 Spreadsheet of microbiological data, 88 5-1 The periodic table of the elements, 93 5-2 Acids and bases on the pH scale, 97 5-3 Structures and names of sample organic molecules, 99 5-4 Nitrogen cycle, 99 B-1 A 2-in. flush nozzle wi
31、ll move only of an 8-in. mains capacity, 108 B-2 Main flushing, 109 Basic Microbiology-AWWA.indb 9 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.Basic Microbiology-AWWA.indb 10 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved
32、.Basic Microbiology 3 rded. xi 4-1 Media comparison, 69 5-1 Valences of various elements, 95 B-1 Flow in gallons per minute from a 2.5-in. diameter nozzle to accomplish 3 ft/sec for the designated pipe diameter, 109 Tables Basic Microbiology-AWWA.indb 11 11/4/13 4:15 PM Copyright 2014 American Water
33、 Works Association. All Rights Reserved.Basic Microbiology-AWWA.indb 12 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.Basic Microbiology 3 rded. xiii This book was written to help those with little or no microbiology educa- tion understand the basic concepts of
34、 the science. Customers and regula- tors expect drinking water plant managers and operators to understand the nature of bacteria, protists, and viruses, which were seldom discussed in the water purification field in the past. For this reason, it is important that water industry personnel become acqu
35、ainted with basic microbiolog- ical concepts to better understand their jobs. Along with its updated core information, this third edition presents the concept of building a progressive microbiology laboratory by includ- ing stream-lined tests for the assessment of aerobic endospore/Cryptospo- ridium
36、 plant performance, and for enumerations of phytoplankton (algae and cyanobacteria), paired with traditional coliform tests, chlorine contact time value considerations, and a method of effective data communication between laboratory personnel and plant operators. Thirty additional photographs have b
37、een added to enhance an under- standing of the text, and colored illustrations accompany many of the sections that discuss individual genera of microorganisms. Also find herein a new appendix that discusses the methods to suc- cessfully decontaminate new mains. I hope that this book successfully ser
38、ves the drinking water managers, operators, laboratory personnel, and others who read it. acknowledgments I thank Des Moines Water Works CEO and General Manager Bill Stowe and other administrative personnel for being supportive of my efforts in developing the microbiology laboratory and in writing t
39、his book. I believe that their support of my educational efforts will also aid other water industry personnel in their water quality endeavors. I also thank the AWW A publishing staff, including Gay Porter De Nileon, Martha Ripley Gray, David Plank, and Melanie Yamamoto; along with proofreader Debor
40、ah J. Lynes and compositor Janice Benight, all of whom helped make this publication possible. D R. H Preface Basic Microbiology-AWWA.indb 13 11/4/13 4:15 PM Copyright 2014 American Water Works Association. All Rights Reserved.Basic Microbiology-AWWA.indb 14 11/4/13 4:15 PM Copyright 2014 American Wa
41、ter Works Association. All Rights Reserved.Chapter 1 1 Introduction Drinking Water Microbiology One of the primary purposes of drinking water treatment is the removal of pathogenic microorganisms. A utility accomplishes this through the application of chemicals, physical filtration, and, occasionall
42、y, biological processes. A utility can most accurately determine its effectiveness of treatment by performing microbiological tests. Chemical and physical methods used to assess basic drinking water quality, such as softening titrations, turbidity testing, particle counting, and chlorine contact tim
43、e (CT) calculations are valuable, but they are not definitive substitutes for direct microbiological determinations. The cor- relation of the data attained from these methods with microbiological con- ditions is empirically derived. In other words, the chemical and physical tests are used to indirec
44、tly assess the microbiological quality of water by relating the results that they generate with data derived from previously conducted studies by research scientists. The Total Coliform Rule developed by US Environmental Protection Agency (USEPA) has been a successful workhorse for several decades,
45、but coliform bacteria are not always ideal surrogates for determining effec- tive viral, protist, or phytoplankton removal. Furthermore, each group of microorganisms presents itself to treatment chemicals and physical filters in a different manner, requiring different assessment methods. Tests such
46、as those for direct Cryptosporidium oocyst detection have been too cumbersome and expensive for practical application in drink- ing water laboratories. Traditional phytoplankton identification and enu- meration methods using Sedgwick-Rafter chambered slides or Utermhl settling chambers require lengt
47、hy processing and microscopic analyses. However, as science has advanced, better opportunities for the determi- nation of microbiological treatment effectiveness have been developed for drinking water utilities. Basic Microbiology-AWWA.indb 1 11/4/13 4:15 PM Copyright 2014 American Water Works Assoc
48、iation. All Rights Reserved.Basic Microbiology 3 rded. 2 Basic Micro Biol ogy for drinking water This third edition of Basic Microbiology for Drinking Water presents a battery of microbiological tests that could be easily adapted for daily use by larger utility laboratories, and for partial or perio
49、dical use by smaller utilities. This battery is especially applicable for utilities that use surface water for their source water. Surface water utilities represent only 22 percent of US community water systems, but these systems serve 70 percent of the people. Application of the streamlined methods presented in this book could potentially advance treatment effectiveness assessments for utilities that serve the majority of drinking water consumers. The microbiological test battery for a progressive laboratory (see chapter 4) would include
