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Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science.

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Presentation on theme: "Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science."— Presentation transcript:

1 Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science

2 Study sites Pauoa Bay (W. Hawaii) Hilo Bay (E. Hawaii) 1 km 100 m Leeward side Low rainfall Ground water plume Windward side High rainfall Groundwater plume(s) SGD Rivers Pauoa Bay Hilo Bay Hawaii Island

3 EPSCoR MARINE sites, including Pauoa Bay Marine ecosystem response to environmental variation, such as climate change (atmospheric-oceanic) and submarine groundwater discharge Kīholo Plot Kaloko Plot Hawaii Island Kīholo Bay Pauoa Bay

4 Why examine phytoplankton responses to coastal hydrology? A complex, fast-responding microbial assemblage – many types, different needs, different fates –A good indicator Biomass serves as a nutrient vector (down); productivity; HABs Size structure responds to environmental forcing Size structure can determine the fate of phytoplankton -sinking rate -grazing losses Environment Phytoplankton Ecosystem

5 Flow Cytometry What can a flow cytometer do? 1.Count cells (up to 10,000 per second) 2.Optically classify each cell, one at a time, 10,000 per second Can distinguish phytoplankton from other cells 3.Quantify optical properties of cells and generate statistics Side and forward scatter – indicator of cell size and shape Fluorescence (green, orange, red) – indicator of pigmentation Flow Cytometry is an important tool for our research

6 PMT Dichroic Filters Bandpass Filters Flow Cytometry Optics Laser 1 2 3 4 Flow cell J. Paul Robinson, Purdue University Light scatter detectors

7 Calibration with size standards (latex beads) FSC-H and FSC-A poorly resolved 1 & 2 micron beads SSC-H and SSC-A didnt have this problem The machine needs to be calibrated against phytoplankton for absolute sizing

8 Flow Cytometry: a monitoring tool for Hawaiian waters Exploring two parameters from each sample –Total phytoplankton (cells / mL) Sum events within regions –Avg ESD Weighted average of cell diameter Avg ESD = (F 1 *ESD 1 )+ (F 2 *ESD 2 )+ (F 3 *ESD 3 )+ (F 4 *ESD 4 ) F = fractional count for the region ESD = geo. mean ESD for that region Red Fluorescence Cell Size Bacteria are analyzed similarly in each sample by applying a fluorescent stain

9 Phytoplankton in SGD plumes of West Hawaii Island Counting and optical characterization –Abundance, size and pigmentation Size Chlorophyll fluorescence Typical off-shore (surface) Kaloko: near-shore (surface) Kiholo: near-shore (surface) Groundwater-influenced waters

10 Depth profile of salinity and phytoplankton (determined by flow cytometry) from Station 14 on July 27, 2010. Surface Bottom FSC-A = forward scatter area (size) FL3A = levels of red fluorescence Middle Larger phytoplankton are constrained to surface plume (Kaloko Bay)

11 Euglena-like cells Synechococcus Euglenoid blooms of West Hawaii (green / black water) ~70 m 1.A Euglenoid causes the dark water -Not toxic 2.Its a large cell with potential to sediment, but the blooms are localized 3.Reduced circulation set favorable conditions for the bloom to occur We now have a tool for monitoring this Euglenoid to learn better how it fits in to the ecosystem

12 Phytoplankton Characteristics at Different Sites of West Hawaii Euglenoid bloom Kīholo Plot Kaloko Plot Pauoa Bay

13 Conclusion Research – Do phytoplankton respond to nutrients in SGD? Approach: Flow cytometry –A tool to address this question (numbers and size of phytoplankton) –Developing a broadly applicable monitoring tool

14 Real-time Continuous Water Quality Buoy Deployed in Hilo Bay Western portion of Hilo Harbor Moored with >100 lbs steel Want to capture storm / base flow conditions April 7, 2010 Still there! Tides FW SGD Buoy for Kiholo Bay is ready to go!

15 Solar powered Data logger / cell phone modem USCG beacon bumper float waterline probes ballast / battery Mooring attachment ~6 feet YSI EMM 68 Hilo Bay WQ Buoy

16 Hilo Bay Buoy Data: Salinity and Wailuku Flow

17 Apx one day lag between buoy and stream gauge Peak discharge Minimum salinity

18 Storm Surges: Turbidity in Hilo Bay

19 Turbidity subsides before salinity recovers Nov 7, 2010 – Hilo Bay From Rt 19 overlook Hamakua coast

20 Hilo Bay Chl a and Salinity

21 Storms flush Chl a from Hilo Bay Recovery of Chl a? Vertical migrations?

22 Conclusions Many tools in place at UH Hilo to examine hydrologic influences on coastal phytoplankton / microbes –Developing indicator tools Understanding the influence of groundwater and surface water is a key research drive Linkages to terrestrial biomes is key

23 Acknowledgements NSF EPSCoR III (UH System) –Kiholo / Kaloko Fairmont Orchid Green Committee –Pauoa Bay NSF EPSCoR II (UH System) –Hilo Bay Tracy Wiegner (UHH Marine Science) Students: Judy Walker, Erick Johnson, Rebecca Most, Gillian Wysock, Ambyr Mokiao-Lee, Javez Mooteb, John Burns (Corals)

24 Conclusions East Hawaii – Ground water nutrients likely fuel productivity; Storm flows flush Hilo Bay, reduce productivity. –How has the biology adapted to these conditions? West Hawaii – Phytoplankton differ between SGD / non- SGD plume areas Physical conditions limit use of nutrients

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