United States Environmental Protection Agency Office of Water Office of Environmental Information Washington, DC EPA-843-R10-002 National Wetland Condition Assessment Laboratory Operations Manual National Wetlands Condition Assessment Laboratory Operations Manual Page ii National Wetlands Condition Assessment Laboratory Operations Manual Page iii TABLE OF CONTENTS 1. INTRODUCTION ................................................................................................................. 1 1.1 Form Logistics ............................................................................................................ 2 1.2 Tracking Samples ....................................................................................................... 2 1.3 Sending Resultant Data Forms ................................................................................... 3 1.4 References ................................................................................................................. 3 2. WATER CHEMISTRY .......................................................................................................... 4 2.1 Introduction to Indicator .............................................................................................. 4 2.2 Parameters for the NWCA .......................................................................................... 4 2.3 Performance-based Methods ...................................................................................... 5 2.4 Sample Processing and Preservation ......................................................................... 5 2.5 Quality Assurance/Quality Control (QA/QC) Procedures ............................................ 7 2.5.1 Internal QA/QC Procedures .................................................................................... 8 2.6 References ................................................................................................................. 8 3. ALGAE ................................................................................................................................. 9 3.1 Introduction to Indicator .............................................................................................. 9 3.1.1 References ............................................................................................................11 3.2 Subsampling ..............................................................................................................11 3.2.1 Safety Precautions .................................................................................................11 3.2.2 Recommended Apparatus/Equipment ...................................................................12 3.2.3 Methods .................................................................................................................13 3.2.4 Quality Assurance/Quality Control .........................................................................13 3.2.5 References ............................................................................................................14 3.3 Diatom Cleaning ........................................................................................................14 3.3.1 Definitions ..............................................................................................................14 3.3.2 Safety Precautions .................................................................................................15 3.3.3 Recommended Apparatus/Equipment ...................................................................15 3.3.4 Methods .................................................................................................................15 3.3.5 Quality Assurance/ Quality Control ........................................................................17 3.3.6 References ............................................................................................................17 3.4 Preparation of Diatom Slides using Naphrax™ Mounting Medium .............................17 3.4.1 Safety Precautions .................................................................................................18 3.4.2 Recommended Apparatus/ Equipment ..................................................................18 3.4.3 Methods .................................................................................................................19 3.4.4 Quality Assurance/ Quality Control ........................................................................23 3.4.5 References ............................................................................................................23 3.5 Analysis of Diatoms ...................................................................................................24 3.5.1 Recommended Apparatus/ Equipment ..................................................................24 National Wetlands Condition Assessment Laboratory Operations Manual Page iv 3.5.2 Methods .................................................................................................................24 3.5.3 Quality Assurance/ Quality Control ........................................................................26 3.5.4 References ............................................................................................................26 3.6 Analysis of All Algae ..................................................................................................28 3.6.1 Definitions ..............................................................................................................28 3.6.2 Recommended Apparatus/ Equipment ..................................................................29 3.6.3 Methods .................................................................................................................29 3.6.4 Calculations and Reporting ....................................................................................32 3.6.5 Quality Assurance/ Quality Control ........................................................................33 3.6.6 References ............................................................................................................33 3.7 Phytoplankton Chlorophyll a ......................................................................................33 3.7.1 Summary of Method ..............................................................................................35 3.7.2 Definitions ..............................................................................................................35 3.7.3 Interferences ..........................................................................................................35 3.7.4 Safety Precautions .................................................................................................36 3.7.5 Equipment and Supplies ........................................................................................36 3.7.6 Reagents and Standards .......................................................................................37 3.7.7 Sample Collection, Preservation and Storage ........................................................38 3.7.8 Calibration and Standardization .............................................................................39 3.7.9 Procedure ..............................................................................................................40 3.7.10 Data Analysis and Calculations ..........................................................................41 3.7.11 Method Performance .........................................................................................42 3.7.12 Pollution Prevention ...........................................................................................42 3.7.13 Waste Management ...........................................................................................42 3.7.14 Quality Control ...................................................................................................43 3.7.15 References ........................................................................................................43 3.8 Microtiter Plate Enzyme-Linked Immuno-Sorbent Assay (ALISA) for Microcystin .....44 3.8.1 Definitions ..............................................................................................................44 3.8.2 Safety Precautions .................................................................................................45 3.8.3 Equipment and Supplies ........................................................................................45 3.8.4 Procedures ............................................................................................................46 3.8.5 Quality Control .......................................................................................................51 3.8.6 References ............................................................................................................52 3.9 Taxonomic Standards Protocols ................................................................................52 3.9.1 Images ...................................................................................................................54 3.9.2 Table of References ..............................................................................................55 3.9.3 Taxonomic Harmonization .....................................................................................55 3.9.4 Final Products ........................................................................................................56