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The Existing Literature in Connection with the Bacteriology

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The Existing Literature in Connection with the Bacteriology ‘The existing literature in connection with the Bacteriology of Water is so extensive, that we venture to think an urgent necessity has arisen for a work in which this subject is specially discussed.’ Percy Frankland, 1894 Micro-organisms in Water – Their significance, identification and removal Promoter Prof. dr. ir. Nico Boon Department of Biochemical and Microbial Technology, Faculty of Bioscience Engineering, Ghent University, Belgium Co-promoter Dr. Frederik Hammes Department of Environmental Microbiology, Eawag: Swiss Federal Institute of Aquatic Science and Technology, Switzerland Members of the examination committee Dr. Ameet Pinto School of Engineering, University of Glasgow, United Kingdom Ing. Sabine Kreps VITO: Flemish Institute for Technological Research, Belgium Prof. dr. ir. Mieke Uyttendaele Department of Food Safety and Food Quality, Faculty of Bioscience Engineering, Ghent University, Belgium Prof. dr. ir. Arne Verliefde Department of Applied Analytical and Physical Chemistry, Faculty of Bioscience Engineering, Ghent University, Belgium Dean Faculty of Bioscience Engineering Prof. dr. ir. Guido Van Huylenbroeck Rector Ghent University Prof. dr. Anne De Paepe GROWTH AND FLOW CYTOMETRIC MONITORING OF BACTERIA IN DRINKING WATER IR. SAM VAN NEVEL Thesis submitted in fulfillment of the requirements for the degree of Doctor (PhD) in Applied Biological Sciences: Environmental Technology Dutch translation of the title Groei en flowcytometrische monitoring van bacteriën in drinkwater Funding This work was supported by the Research Foundation – Flanders (Fonds voor Wetenschappelijk Onderzoek (FWO) Vlaanderen), project grant no. G.0808.10N. Cover illustration Cover model: Sanne Van Nevel Flow cytometry scheme Network map FARYS © Timternet Infiltration pond St-André IWVA, Oostduinkerke Van Nevel S. (2014) Growth and flow cytometric monitoring of bacteria in drinking water. PhD thesis, Ghent University, Belgium. ISBN: 978-90-5989-759-5 The author and the promoters give the authorisation to consult and to copy parts of this work for personal use only. Every other use is subject to the copyright laws. Permission to reproduce any material contained in this work should be obtained from the author. NOTATION INDEX AAS Atomic absorbance spectrophotometry AOC Assimilable organic carbon API Analytical profile index ATP Adenosine triphospate AWWA American Water Works Association BDOC Biodegradable dissolved organic carbon BOD Biological oxygen demand CARD-FISH Catalysed reporter deposition fluorescent in situ hybridisation CFU Colony forming units CODt Total chemical oxygen demand cTEP Colloidal transparent exopolymer particles DALY Disability-adjusted life years DGGE Denaturing gradient gel electrophoresis DMSO Dimethylsulfoxide DOC Dissolved organic carbon DW Drinking water DWC Drinking water community DWDS Drinking water distribution system EFM Epifluorescence microscopy EPA Environmental Protection Agency EPS Extracellular polymeric substance FCM Flow cytometry FDA Ficher discriminant analysis FISH Fluorescent in situ hybridisation FL1, FL2,… Fluorescence detectors of a flow cytometer, numbered starting from the lowest wavelength FSC Forward scatter detector i Notation index fx Calibration factor for expressing alcian blue absorbance as xanthan gum HPC Heterotrophic plate count IC Indigenous community ICC Intact cell counts LOD Limit of detection LOQ Limit of quantification MALDI-TOF Matrix-assisted laser desorption ionization time-of-flight MAP Microbially available phosphorus MBR Membrane bioreactor MF Membrane filtration MPN Most probable number MTF Multiple tube fermentation NF Nanofiltration NGS Next-generation sequencing PACl Poly aluminium chloride PC Principal components PCA Principal component analysis PCR Polymerase chain reaction PI Propidium iodide PMA Propidium monazide pTEP Particular transparent exopolymer particles qPCR Quantitative real-time polymerase chain reaction R2a Reasoner’s 2a (agar) RO Reverse osmosis RSD Relative standard deviation SCADA Supervisory control and data acquisition SG SYBR Green I SPR Surface plasmon resonance SSC Side scatter detector STP Sewage treatment plant ii Notation index SW Surface water SWC Surface water community TCC Total cell counts TEP Transparent exopolymer particles TGGE Temperature gradient gel electrophoresis TOC Total organic carbon T-RFLP Terminal restriction fragment length polymorphism UF Ultrafiltration VBNC Viable but non-culturable WHO World Health Organisation WPC Water plate count (agar) iii TABLE OF CONTENTS CHAPTER 1 GENERAL INTRODUCTION ON BACTERIOLOGY IN DRINKING WATER DISTRIBUTION SYSTEMS (DWDS) ......................................................................................................................................... 1 1. DESCRIPTION OF BACTERIA IN DWDS .............................................................................................. 1 1.1 BACTERIAL PRESENCE IN FOUR PHASES IN DRINKING WATER ............................................................................... 1 1.2 COMMONLY FOUND BACTERIA IN DRINKING WATER ......................................................................................... 2 1.3 CONSEQUENCES ........................................................................................................................................ 4 1.3.1 Health-based implications ..................................................................................................... 4 1.3.2 Network integrity and hydrology ........................................................................................... 6 1.3.3 Dissatisfaction and customer complaints .............................................................................. 6 2. WHAT AFFECTS BACTERIAL GROWTH IN DWDS? ............................................................................... 7 2.1 PRESENCE OF NUTRIENTS ............................................................................................................................ 7 2.2 BACTERIAL COMPETITION ............................................................................................................................ 8 2.3 OTHER CONDITIONS ................................................................................................................................... 8 BOX 1-1: THE POSSIBLE ROLE OF TRANSPARENT EXOPOLYMER PARTICLES (TEP) AS BIOFILM INITIATOR. ............................. 10 2.4 MEASURES TO LIMIT BACTERIAL GROWTH .................................................................................................... 11 3. NON-SELECTIVE DETECTION OF BACTERIA IN DWDS ......................................................................... 14 3.1 ENUMERATION OF BACTERIA BY HPC: STILL THE GOLD STANDARD ..................................................................... 14 3.1.1 130 years of development.................................................................................................... 14 3.1.2 What is HPC showing? ......................................................................................................... 15 3.1.3 What is HPC missing? .......................................................................................................... 16 3.2 FLOW CYTOMETRIC CELL COUNTS ............................................................................................................... 19 3.3 ATP-QUANTIFICATION ............................................................................................................................. 20 BOX 1-2: FLOW CYTOMETRY (FCM) – UNDERSTANDING CELL COUNTING ................................................................... 21 BOX 1-3: FLOW CYTOMETRIC (FCM) FINGERPRINTING ............................................................................................ 29 4. SELECTIVE DETECTION.................................................................................................................. 34 4.1 CULTIVATION-DEPENDENT METHODS .......................................................................................................... 34 BOX 1-4: QUANTITATIVE MICROBIAL RISK ASSESSMENT (QMRA) .............................................................................. 39 4.2 CULTIVATION-INDEPENDENT METHODS ....................................................................................................... 42 5. OBJECTIVES AND OVERVIEW OF THE RESEARCH ................................................................................ 44 CHAPTER 2 TRANSPARENT EXOPOLYMER PARTICLE REMOVAL IN DIFFERENT DRINKING WATER PRODUCTION CENTERS ............................................................................................................................ 51 ABSTRACT ............................................................................................................................................ 51 1. INTRODUCTION .......................................................................................................................... 52 v Table of contents 2. MATERIALS AND METHODS .......................................................................................................... 55 2.1 SAMPLING ............................................................................................................................................. 55 2.2 TEP MEASUREMENT ................................................................................................................................ 55 2.3 ANALYTICAL METHODS ............................................................................................................................
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