Golden Alga Workshop Summary Report - Texas Parks & Wildlife ...

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How to design and test a probe * Amass data bases from rRNA sequences * use ARB program to design probe * check probe for possible matches in RDP and Genbank * test specificity in dot blot (DIG-labelled probe) and in situ (FITC or CY3 labelled probe) tests PWD RP T3200-1203 * final check with flow cytometer
EUK 1209R CHLO 02 PRYM 02 PELA 01 PELA 02 PWD RP T3200-1203
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Golden Alga (Prymnesium parvum) Wor
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Golden Alga Workshop Agenda October
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Golden Alga Workshop Table of Conte
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Golden Alga Workshop Background Inf
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The TPWD “GOLD” Team Technical
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While There Is Much We Do Not Know
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Detection Early Simple Basic Resear
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Historical Review of Golden Alga (P
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Prymnesium parvum PWD RP T3200-1203
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Prymnesium parvum in Texas: 28 eve
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Five Texas River Basins Impacted C
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Number and Value of Fish Killed 198
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1 1 5 5 6 6 14 13 15 12 14 13 15 12
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Month Golden Alga Fish Kills Began
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Number of Fish Killed Brazos Basin
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We Want to Solve the Problem Factor
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What We Have Done - continued Repor
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Special Thanks To The Following: We
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Overview of Texas Hatchery Manageme
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Dundee State Fish Hatchery • Febr
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Dundee State Fish Hatchery • Caus
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TPWD Inland Fisheries P. parvum Tas
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Monitoring P. parvum densities •
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Monitoring Toxin Levels • Bioassa
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Monitoring of P. parvum • Needs -
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Physical Treatments Χ Sonication (
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Χ X Chemical Treatments Hydrogen p
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Treatments - Chemical • Ammonium
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Treatments - Chemical • Cutrine
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Achievements • Development of hat
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Phylogeny, life history, autecology
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Overview • morphology - what it l
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Morphology of P. parvum haptonema f
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Prymnesium species Species Habitat
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Haplo-diploid life cycle P. parvum
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Life cycle: mating experiment 2n +
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Possible life cycle for P. parvum H
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Optimum and tolerance for growth Pa
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Mixotrophy • P. parvum can ingest
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Toxins • proteolipids (Ulizur & S
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Occurrence of harmful Typical habit
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What can be done? • Establish a m
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Prymnesium parvum: An overview and
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Presently recognized Prymnesium spe
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Bloom History • Within a decade o
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PWD RP T3200-1203
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Dying Shad - Texas - courtesy of C.
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PWD RP T3200-1203
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Dead fish at a dam site in Texas -
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Fish showing hemmoragic areas from
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Toxins: (Lethal Cocktail) There is
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How easily can they be identified f
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PWD RP T3200-1203 Body Scales of P.
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Fluorescent labeled P. parvum cell
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Priorities: Cells: • accurate and
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Analysis of Prymnesium parvum bloom
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Overview • Questions Identified b
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Approach - proposed studies • Syn
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Lake Whitney Stations Site P11 Site
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80000 Lake Whitney 2003 P. parvum a
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Feb-April 2003 Lake Whitney: surfac
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Lake Bioassay Methods • Acclimate
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Lake Whitney Bioassay Sites Site P1
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Lake Whitney Nutrient Bioassay 4/29
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Original Questions • What factors
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Next steps? • Continue paired-res
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DY III Media Stock Addtions Nutrien
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Kill your enemies and eat them: the
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5 µm Killed salmons in aquaculture
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When will the first dead bathing gu
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PWD RP T3200-1203
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Toxicity in Chrysochromulina polyle
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HE 50 (10 6 cells ml -1 ) Prymnesiu
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Effect of increasing P-deficent P.
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Ichthyotoxic species Prymnesin-2 H
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☺ Before PWD RP T3200-1203
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Effect of Prymnesium parvum filtrat
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Skovgaard and Hansen, 2003, L & O,
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Ciliates Flagellates Diatoms cell l
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Low Prymnesium parvum cell numbers
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6 Bacterivory in P. parvum Ingested
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P. parvum phagotrophy (prey ingesti
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% P. parvum feeding cells and lysed
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Skovgaard and Hansen, 2003, L & O,
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Oxyrrhis marina cell lysis in the p
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Ingestion of Oxyrrhis after 2 and 6
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P-deficient P. parvum ingesting blo
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Conclusions •Toxicity is the key
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C. helgolandicus pellets •Picture
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Laboratory Studies on Prymnesium -
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News Release • May 28, 2002 - mas
Page 187 and 188: Fish Kill at Prewitt Reservoir PWD
Page 189 and 190: Pelicans putative of transport agen
Page 191 and 192: Objectives of this Presentation •
Page 193 and 194: Prymnesium Strains in Culture • U
Page 195 and 196: Structure of Prymnesium PWD RP T320
Page 197 and 198: PWD RP T3200-1203
Page 199 and 200: Scales Haptonema PWD RP T3200-1203
Page 201 and 202: Cyst Formation • A variety of con
Page 203 and 204: Growth Observations 0n Prymnesium s
Page 205 and 206: Prymnesium Twin Buttes Lake, Wyomin
Page 207 and 208: Hemolytic Bioassay Procedures • 5
Page 209 and 210: Hemolysis Bioassay • Alga Medium
Page 211 and 212: Mixotrophy Studies • Several phyt
Page 213 and 214: Chloroplast Endoplasmic Reticulum N
Page 215 and 216: Adaptive Advantage of Chloroplast E
Page 217 and 218: 10 µm Kathablepharis sp. Courtesy
Page 219 and 220: Chrysochromulina parva Long, uncoil
Page 221 and 222: Ingested Chrysochromulina & some ba
Page 223 and 224: Return to Prewitt • Prymnesium pa
Page 225 and 226: Prewitt Reservoir Fact • Five day
Page 227 and 228: Prewitt Reservoir Explanation • P
Page 229 and 230: Prewitt Mystery Solved? • Campylo
Page 231 and 232: Future Prymnesium Problems in Wyomi
Page 233 and 234: Rapid Tests for the Detection of Pr
Page 235 and 236: Why rRNA probes? * universally foun
Page 237: PWD RP T3200-1203
Page 241 and 242: Change from PFA to ETOH Saline Chan
Page 243 and 244: Analysis of Alexandrium strains fro
Page 245 and 246: PWD RP T3200-1203
Page 247 and 248: Harmful Algal Blooms Noctiluca Kare
Page 249 and 250: PWD RP T3200-1203 Coccolithus pelag
Page 251 and 252: Application & detection methods for
Page 253 and 254: Conclusions • Specific probes cou
Page 255 and 256: 1. ChemScanRDI, a laser based syste
Page 257 and 258: ChemScanRDI combines fluorescent ce
Page 259 and 260: Electrochemical detection of toxic
Page 261 and 262: Electrochemical Detection of rRNA f
Page 263 and 264: Comparison of cell counts with elec
Page 265 and 266: Long term stability of treated sens
Page 267 and 268: Development of DNA-microarrays for
Page 269 and 270: Scheme of a DNA-Chip Experiment Flu
Page 271 and 272: Monitoring Phytoplankton compositio
Page 273 and 274: EUK 1209 CHLO01 Localization of the
Page 275 and 276: Identification of Phytoplankton on
Page 277 and 278: DNA CHIP Clone librairy Dec 2000 1
Page 279 and 280: Research Needs • Monitoring of to
Page 281 and 282: Rapid Tests for the Detection of Ha
Page 283 and 284: abs. units 1.0 0.8 0.6 0.4 0.2 y =
Page 285 and 286: Prymnesium parvumRL10 % lysis 100 8
Page 287 and 288: PWD RP T3200-1203 Allelochemical ef
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Controlling Harmful Algal Blooms Do
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Management of harmful algae • Pre
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Chemical control of freshwater alga
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•This (small) reservoir had a lon
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Chemical flocculants - Phosphorus C
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PWD RP T3200-1203 from: River Scien
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Biological Control - Viruses Aureoc
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Viruses for HAB species Target spec
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Biological Control - Bacterial path
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Biological Control - Grazers Target
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Clay control of HAB species clay mi
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Variable removal ability of domesti
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PWD RP T3200-1203 Source: Sengco et
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Flume studies, WHOI PWD RP T3200-12
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Brevetoxin analysis cell concentrat
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Impacts - Benthic fauna Archambault
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Future directions: Cell removal, se
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When the cells reached exponential
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Conclusions Kalmar Experiments Phos
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Experiments at Woods Hole (in colla
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Recent work in Kalmar (data from J.
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Conclusions - control of Prymnesium
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A review of fish-killing microalgae
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Toxic/harmful microalgae • dinofl
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PWD RP T3200-1203 Fish kills
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Ichthyotoxic species • Karenia br
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FMRI,FWC Exposure routes • gills
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Gymnodinium pulchellum (brevetoxins
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Reef fish disease - Caribbean, Flor
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Diatoms • physical damage to gill
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Aquaculture in Israel and Prymnesiu
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Prymnesium parvum from Israel PWD R
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Prymnesium parvum blooms in Israel
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From Sarig 1971 Prymnesium parvum t
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From Sarig 1971 Mode of toxin actio
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Fish bioassay • dependence of tox
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Relationship # toxin concentration
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From Sarig 1971 Management of Prymn
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Management strategies • prevent b
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How to Use the Past to Plan for the
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How can studies on other harmful al
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Task Force report identified 7 HAB
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A 5-year federal, state, academic,
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ECONOMIC IMPACT In the 1970s, two r
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What is Needed Based on What is Kno
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Golden Alga Workshop Notes: Present
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facilities was a problem in North C
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Korean fisheries are a $1 billion i
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Friday October 24, 2003 Panel Discu
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• Design a sampling program with
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What are your recommendations for i
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• Rapid progress is made where th
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• Maintain a Golden Alga web site
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ChemScanRDI (a laser based system t
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• DNA-microarrays for monitoring
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• Reliable economic statistics fo
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• Be proactive; get messages out
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Golden Alga Workshop Attendees Firs
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Golden Alga Workshop Attendees, con
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Literature Review of the Microalga
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Oued Mellah Reservoir in Morocco oc
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the metal ions Fe, Zn, Mo, Cu or Co
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with the ability to utilize organic
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Figure 1. Prymnesin-1 (1) and Prymn
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Temperature Shilo and Ashner (1953)
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Glycerol Glycerol was found to incr
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since it seems that nitrogen and ph
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(Haptophyta) in semi-continuous cul
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Ulitzer, S. and M. Shilo. 1966. Mod
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