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Manual for Phytoplankton Sampling and Analysis in the Black Sea

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Manual for Phytoplankton Sampling and Analysis in the Black Sea Manual for Phytoplankton Sampling and Analysis in the Black Sea Dr. Snejana Moncheva Dr. Bill Parr Institute of Oceanology, Bulgarian UNDP-GEF Black Sea Ecosystem Academy of Sciences, Recovery Project Varna, 9000, Dolmabahce Sarayi, II. Hareket P.O.Box 152 Kosku 80680 Besiktas, Bulgaria Istanbul - TURKEY Updated June 2010 2 Table of Contents 1. INTRODUCTION........................................................................................................ 5 1.1 Basic documents used............................................................................................... 1.2 Phytoplankton – definition and rationale .............................................................. 1.3 The main objectives of phytoplankton community analysis ........................... 1.4 Phytoplankton communities in the Black Sea ..................................................... 2. SAMPLING ................................................................................................................. 9 2.1 Site selection................................................................................................................. 2.2 Depth ............................................................................................................................... 2.3 Frequency and seasonality ....................................................................................... 2.4 Algal Blooms................................................................................................................. 2.4.1 Phytoplankton bloom detection in the field.................................................. 2.4.2 Phytoplankton bloom measures...................................................................... 2.4.3 Evaluation forms .................................................................................................. 2.5 Equipment.................................................................................................................. 11 2.6 Methodology.................................................................................................................. 2.6.1 Qualitative sampling ........................................................................................... 2.6.2 Quantitative sampling......................................................................................... 3. SAMPLE PRESERVATION AND STORAGE..................................................... 14 3.1 Fixatives/preservatives .............................................................................................. 3.2 Advantages and disadvantages of formaldehyde............................................... 3.2.1 Advantages............................................................................................................ 3.2.2 Disadvantages ...................................................................................................... 3.3 Advantages and disadvantages of Lugol’s........................................................... 3.3.1 Advantages............................................................................................................ 3.3.2 Disadvantages ...................................................................................................... 3.4 Sample Storage ............................................................................................................ 3.4.1 Living samples..................................................................................................... 3.4.2 Preserved samples .............................................................................................. 4. PHYTOPLANKTON ENUMERATION AND DENSITIES.................................. 18 4.1 Equipment...................................................................................................................... 4.2 Sub-sample preparation............................................................................................. 4.2.1 Sample homogenisation .................................................................................... 4.3 Cell counts using an inverted microscope ........................................................... 4.3.1 Surface water removal........................................................................................ 4.3.2 Filtration ................................................................................................................. 4.3.3 Settling chambers................................................................................................ 4.2.3 Counting procedure ............................................................................................ 4.4 Cell counts using a conventional light compound microscope ..................... 4.4.1 Surface water removal........................................................................................ 4.4.2 Filter-transfer-freeze methodology ................................................................. 4.4.3 Centrifugation ....................................................................................................... 4.5 Recording the number of algal cells....................................................................... 4.6 Density calculations.................................................................................................... 5. SEMI-QUANTITATIVE ANALYSIS OF PHYTOPIANKTON SAMPLES........ 32 5.1 Inverted micoscope..................................................................................................... 5.2 Conventional light compound microscope........................................................... 6. TAXONOMIC IDENTIFICATION........................................................................... 33 6.1 Introduction ................................................................................................................... 6.2 Identification books in use within Black sea countries..................................... 6.3 Additional references of recent taxonomic developments .............................. 7. BIOMASS DETERMINATION ............................................................................... 39 7.1 Diatoms........................................................................................................................... 7.2 Calculation of a cell’s reference biovolume ......................................................... 8. RECORDING DATA............................................................................................ 4141 8.1 Bench Sheets ................................................................................................................ 9. PHYTOPLANKTON-RELATED ENVIRONMENTAL STATUS INDICATORS............................................................................................................ 43 9.1 Introduction ................................................................................................................... 9.2 Black Sea-specific phytoplankton-related environmental status indicators ........................................................................................................................... 10. QUALITY CONTROL AND QUALITY ASSURANCE (QC/QA) ...................... 45 10.1 Introduction ................................................................................................................... 10.2 Equipment...................................................................................................................... 10.3 Sampling protocol ....................................................................................................... 10.4 Sample preservation ................................................................................................... 10.5 Sub-sampling – validation of homogenization .................................................... 10.6 Sample analysis ........................................................................................................... 10.6.1 Biovolume/biomass estimation........................................................................ 10.6.2 Phytoplankton enumeration.............................................................................. 10.6.3 Taxonomy............................................................................................................... 10.6.4 Re-analysis ............................................................................................................ 10.6.5 Training and inter-laboratory comparability testing .................................. 10.6.6 Repeatability and reproducibility..................................................................... 10.6.7 Uncertainty............................................................................................................. 10.7 Reporting/data storage........................................................................................... Annex A. STANDARDISED LIST OF BLACK SEA PHYTOPLANKTON TAXA AND REFERENCE BIOVOLUMES ...................................................................... 49 Annex B. CONTROL CHARTS FOR BIOLOGICAL MEASUREMENTS........................ 56 Annex C. MEASUREMENT UNCERTAINTY....................................................................... 58 Annex D. LIST OFPARTICIPANTS…………………………………………………………62 11. REFERENCES CITED IN THE MANUAL ....................................................................... 64 4 This document is intended for use as a guide to harmonize the procedures among the different labs in the Black Sea countries for phytoplankton monitoring. The methods for phytoplankton microscope
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