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

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References Edvardsen, B., Eikrem, W., Green, J.C., Andersen, R.A., Moon-van der Staay, S.Y. & Medlin, L.K. 2000. Phylogenetic reconstructions of the Haptophyta inferred from 18S ribosomal DNA sequences and available morphological data. Phycologia 39 (1), 19-35. Igarashi, T. Satake, M.& Yasumoto, T. 1996. Prymnesin-2: a potent ichthyotoxic and hemolytic glycoside isolated from the red tide algae Prymnesium parvum. J. Am. Chem. Soc. 118: 479-480. Kozakai, H., Oshima, Y.& Yasumoto, T. 1982. Isolation and structural elucidation of hemolysin from the phytoflagellate Prymnesium parvum. Agric. Biol. Chem. 46: 233-236. Larsen, A. 1998. Autecology, toxicity, genetics and life history of Prymnesium parvum and Prymnesium patelliferum (Haptophyta): is a species separation warranted? Dr. Scient. thesis, University of Bergen, Norway. Larsen, A. 1999. Prymnesium parvum and P. patelliferum (Haptophyta) - one species. Phycologia 38:541-543. Larsen, A. & Edvardsen, B. 1998. A study of relative ploidy levels in Prymnesium parvum and P. patelliferum (Haptophyta) analysed by flow cytometry. Phycologia 37(6), 412-424 Larsen, A. Eikrem, W. & Paasche, E. 1993. Growth and toxicity in Prymnesium patelliferum (Haptophyta) isolated from Norwegian waters. Can. J. Bot. 71: 1357-1362. Larsen, A. & Medlin, L.K. 1997. Inter- and intraspecific genetic variation in twelve Prymnesium (Haptophyta) clones. J. Phycol. 33:1007-1015. Tillmann, U. 1998. Phagotrophy by a plastidic haptophyte, Prymnesium patelliferum. Aquat. Microb. Ecol. 14: 155-160. Wang, Y.& Wang, Y. 1992. Biology and classification of Prymnesium saltans. Acta Hydrobiol. Sin. 16: 193-199. PWD RP T3200-1203
Prymnesium parvum: An overview and challenges to our understanding. Major Questions left after 55 years of problems Carmelo R. Tomas, Center for Marine Science, University of North Carolina at Wilmington, 5600 Marvin K. Moss Lane, Wilmington, NC 28409 USA The ichthyotoxic flagellate Prymnesium parvum has historically been a persistent problem in lakes, ponds, rivers and estuaries. In 1937, N. Carter described this species from a brackish pool on the Isle of Wight, England. Within the first decade of its description, this species was identified as causing extensive fish kills in Lake Kinneret (Sea of Galilee), Israel, and in aquaculture ponds. From 1947 through the 1960’s, Israeli scientists performed extensive studies to determine methods of controlling the blooms. The hemolytic nature of the toxin, a general understanding of how the toxin was formed, and the reaction to the fish were described, a standard bioassay was established, and mitigation efforts involving liquid ammonia were tested and offered as a means to control the blooms. This effort, notwithstanding, the blooms of P. parvum persisted and spread elsewhere, and now this species is identified from 14 different countries, spanning Scandinavia, Europe, Africa, Asia, New Zealand, and North and South America. One of the first challenges in dealing with P. parvum is its identification. As a small (> 10 µm) highly motile cell, detection of its flagella and haptonema can be accomplished with brightfield microscopy, but the diagnostic body scales and other features require the use of electron microscopy (TEM and SEM). This is time consuming and requires specialized equipment not practical for routine observations. The blooms of this organism are episodic, appearing overnight and either persisting or disappearing rapidly. Maximum densities often exceed 10 8 cells/liter and once established, P. parvum becomes a persistent feature of the phytoplankton. This last feature suggests a survival stage that needs to be described and detected. Conflicting evidence for conditions accompanying blooms are noted. They occur at low (35 PSU) salinities. This species is photosynthetic but extremely tolerant to low light intensities and can survive in the dark with organic carbon sources. It requires a dark cycle for the production of toxin and has been reported to produce toxins at low (30 °C) temperatures. Nutrients, discussed by others, give an equally confusing picture. Great headway has been accomplished as to the toxins, but it is clear that they are complex, difficult to characterize and detect, and consist of multiple toxins having hemolytic, ichthyotoxic, neurotoxic, reactive oxygen, and other components. Linkages between environmental variables and toxin production are very complicated to discern. Our present knowledge of P. parvum blooms requires a series of efforts to be able to help in understanding their dynamics. Some of the major problems to be confronted are: • Easy, rapid and accurate identification of the P. parvum in natural waters particularly at low cell concentrations. • Detection and distributional mapping of P. parvum resting stages (cysts) for identifying potential bloom initiation points. • Accurate and easy detection of its toxins (kinds and levels) to diagnose problems and to evaluate remediation efforts to relieve bloom effects. • Development of a series of bioassays to guide efforts in defining the fate and effects of the P. parvum toxins (bioassay guided fractionation and mitigation to determine toxin ½ life and potential beneficial uses). • Mitigation and control of the blooms in defined situations via particle removal, cell lysis or other effective agents causing minimal damage to the environment. View the presentation 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
Page 37 and 38: Overview of Texas Hatchery Manageme
Page 39 and 40: Dundee State Fish Hatchery • Febr
Page 41 and 42: Dundee State Fish Hatchery • Caus
Page 43 and 44: TPWD Inland Fisheries P. parvum Tas
Page 47 and 48: Monitoring P. parvum densities •
Page 49 and 50: Monitoring Toxin Levels • Bioassa
Page 51 and 52: Monitoring of P. parvum • Needs -
Page 53 and 54: Physical Treatments Χ Sonication (
Page 55 and 56: Χ X Chemical Treatments Hydrogen p
Page 57 and 58: Treatments - Chemical • Ammonium
Page 59 and 60: Treatments - Chemical • Cutrine
Page 61 and 62: Achievements • Development of hat
Page 65 and 66: Phylogeny, life history, autecology
Page 67 and 68: Overview • morphology - what it l
Page 69 and 70: Morphology of P. parvum haptonema f
Page 71 and 72: Prymnesium species Species Habitat
Page 73 and 74: Haplo-diploid life cycle P. parvum
Page 75 and 76: Life cycle: mating experiment 2n +
Page 77 and 78: Possible life cycle for P. parvum H
Page 79 and 80: Optimum and tolerance for growth Pa
Page 81 and 82: Mixotrophy • P. parvum can ingest
Page 83 and 84: Toxins • proteolipids (Ulizur & S
Page 85 and 86: Occurrence of harmful Typical habit
Page 87: What can be done? • Establish a m
Page 91 and 92: Presently recognized Prymnesium spe
Page 93 and 94: Bloom History • Within a decade o
Page 95 and 96: PWD RP T3200-1203
Page 97 and 98: Dying Shad - Texas - courtesy of C.
Page 99 and 100: PWD RP T3200-1203
Page 101 and 102: Dead fish at a dam site in Texas -
Page 103 and 104: Fish showing hemmoragic areas from
Page 105 and 106: Toxins: (Lethal Cocktail) There is
Page 107 and 108: How easily can they be identified f
Page 109 and 110: PWD RP T3200-1203 Body Scales of P.
Page 111 and 112: Fluorescent labeled P. parvum cell
Page 113 and 114: Priorities: Cells: • accurate and
Page 115 and 116: Analysis of Prymnesium parvum bloom
Page 117 and 118: Overview • Questions Identified b
Page 119 and 120: Approach - proposed studies • Syn
Page 121 and 122: Lake Whitney Stations Site P11 Site
Page 123 and 124: 80000 Lake Whitney 2003 P. parvum a
Page 125 and 126: Feb-April 2003 Lake Whitney: surfac
Page 127 and 128: Lake Bioassay Methods • Acclimate
Page 129 and 130: Lake Whitney Bioassay Sites Site P1
Page 131 and 132: Lake Whitney Nutrient Bioassay 4/29
Page 133 and 134: Original Questions • What factors
Page 135 and 136: Next steps? • Continue paired-res
Page 137 and 138: 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
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Fish Kill at Prewitt Reservoir PWD
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Pelicans putative of transport agen
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Objectives of this Presentation •
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Prymnesium Strains in Culture • U
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Structure of Prymnesium PWD RP T320
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PWD RP T3200-1203
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Scales Haptonema PWD RP T3200-1203
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Cyst Formation • A variety of con
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Growth Observations 0n Prymnesium s
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Prymnesium Twin Buttes Lake, Wyomin
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Hemolytic Bioassay Procedures • 5
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Hemolysis Bioassay • Alga Medium
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Mixotrophy Studies • Several phyt
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Chloroplast Endoplasmic Reticulum N
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Adaptive Advantage of Chloroplast E
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10 µm Kathablepharis sp. Courtesy
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Chrysochromulina parva Long, uncoil
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Ingested Chrysochromulina & some ba
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Return to Prewitt • Prymnesium pa
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Prewitt Reservoir Fact • Five day
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Prewitt Reservoir Explanation • P
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Prewitt Mystery Solved? • Campylo
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Future Prymnesium Problems in Wyomi
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Rapid Tests for the Detection of Pr
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Why rRNA probes? * universally foun
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PWD RP T3200-1203
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EUK 1209R CHLO 02 PRYM 02 PELA 01 P
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Change from PFA to ETOH Saline Chan
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Analysis of Alexandrium strains fro
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PWD RP T3200-1203
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Harmful Algal Blooms Noctiluca Kare
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PWD RP T3200-1203 Coccolithus pelag
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Application & detection methods for
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Conclusions • Specific probes cou
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1. ChemScanRDI, a laser based syste
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ChemScanRDI combines fluorescent ce
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Electrochemical detection of toxic
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Electrochemical Detection of rRNA f
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Comparison of cell counts with elec
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Long term stability of treated sens
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Development of DNA-microarrays for
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Scheme of a DNA-Chip Experiment Flu
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Monitoring Phytoplankton compositio
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EUK 1209 CHLO01 Localization of the
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Identification of Phytoplankton on
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DNA CHIP Clone librairy Dec 2000 1
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Research Needs • Monitoring of to
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Rapid Tests for the Detection of Ha
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abs. units 1.0 0.8 0.6 0.4 0.2 y =
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Prymnesium parvumRL10 % lysis 100 8
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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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