flow cytometric analyses and rtcm of water quality around hawaii island jason e. adolf and judy k....
TRANSCRIPT
![Page 1: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/1.jpg)
Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island
Jason E. Adolf and Judy K. WalkerUHH Marine Science
![Page 2: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/2.jpg)
Study sites
Pauoa Bay(W. Hawaii)
Hilo Bay(E. Hawaii)
1 km100 m
Leeward sideLow rainfallGround water plume Windward side
High rainfallGroundwater plume(s)
SGD
SGD
Rivers
Pauoa Bay
Hilo Bay
Hawai‘i Island
![Page 3: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/3.jpg)
EPSCoR MARINE sites, including Pauoa BayMarine ecosystem response to environmental variation, such as climate change (atmospheric-oceanic) and submarine groundwater discharge
Kīholo Plot
Kaloko Plot
Hawai‘i Island
Kīholo Bay
Pauoa Bay
![Page 4: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/4.jpg)
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
![Page 5: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/5.jpg)
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 cells3. 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
![Page 6: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/6.jpg)
PMT
PMT
PMT
PMT
DichroicFilters
BandpassFilters
Flow Cytometry Optics
Laser
1
2
3
4
Flow cell
J. Paul Robinson, Purdue University
Light scatterdetectors
![Page 7: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/7.jpg)
Calibration with size standards(latex beads)
FSC-H and FSC-A poorly resolved 1 & 2 micron beadsSSC-H and SSC-A didn’t have this problem
The machine needs to be calibrated against phytoplankton for absolute sizing
![Page 8: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/8.jpg)
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 = (F1*ESD1)+ (F2*ESD2)+ (F3*ESD3)+ (F4*ESD4)
F = fractional count for the regionESD = geo. mean ESD for that region
Red
Flu
ores
cenc
e
Cell Size
Bacteria are analyzed similarly in each sample by applying a fluorescent stain
![Page 9: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/9.jpg)
Phytoplankton in SGD plumes ofWest Hawaii Island
• Counting and optical characterization– Abundance, size and pigmentation
Size
Chl
orop
hyll
fluor
esce
nce
Typical off-shore(surface)
Kaloko: near-shore(surface)
Kiholo: near-shore(surface)
Groundwater-influenced waters
![Page 10: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/10.jpg)
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
33 34 35 36 37
Salinity (ppt)
Dep
th (
m)
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)
![Page 11: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/11.jpg)
Euglena-likecells
Synechococcus
Euglenoid blooms of West Hawaii (green / black water)
~70 mm
1. A Euglenoid causes the dark water-Not toxic
2. It’s 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
![Page 12: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/12.jpg)
Phytoplankton Characteristics atDifferent Sites of West Hawaii
Euglenoid bloom
Kīholo Plot
Kaloko Plot
Pauoa Bay
![Page 13: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/13.jpg)
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
![Page 14: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/14.jpg)
Real-time Continuous Water Quality Buoy Deployed in Hilo Bay
Western portion ofHilo Harbor
Moored with >100 lbs steel
Want to capture storm / base flow conditions
April 7, 2010Still there!
Tides
FWSGD
Buoy for Kiholo Bay is ready to go!
![Page 15: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/15.jpg)
Solar powered
Data logger / cell phone modemUSCG beacon
bumper
floatwaterline
probes
ballast /battery
Mooringattachment
~6 feet
YSI EMM 68
Hilo BayWQ Buoy
![Page 16: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/16.jpg)
Hilo Bay Buoy Data:Salinity and Wailuku Flow
![Page 17: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/17.jpg)
Apx one day lag betweenbuoy and stream gauge
Peak dischargeMinimum salinity
![Page 18: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/18.jpg)
Storm Surges: Turbidity in Hilo Bay
![Page 19: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/19.jpg)
Turbidity subsides before salinity ‘recovers’
Nov 7, 2010 – Hilo Bay From Rt 19 overlook
Hamakua coast
![Page 20: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/20.jpg)
Hilo Bay Chl a and Salinity
![Page 21: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/21.jpg)
Storms flush Chl a from Hilo Bay
Recovery of Chl a?
Vertical migrations?
![Page 22: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/22.jpg)
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
![Page 23: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/23.jpg)
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)
![Page 24: Flow Cytometric Analyses and RTCM of Water Quality around Hawaii Island Jason E. Adolf and Judy K. Walker UHH Marine Science](https://reader037.vdocuments.us/reader037/viewer/2022102923/55182f09550346ac318b4f60/html5/thumbnails/24.jpg)
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