Southwest Missouri State University An Overview of Harmful Algal Blooms (HABs); Diversity and Dynamics of Toxic Algae Russell G. Rhodes Department of Biology Southwest Missouri State University

My Interests in HABs 1. Resource for MDC 1989- Notified by Missouri Department of Conservation of a Fish Kill by “Bluegreen algae” Identified presumptive agent as a dinoflagellate 1990- A graduate student worked out the life history 1991- Published a paper about the organism Fields, S. D. and R. G. Rhodes. 1991. Ingestion and retention of Chroomonas spp. (Cryptophyceae) by Gymnodinium acidotum (Dinophyceae). J. Phycology 27:525-529.

2. Resource for Department of Natural Resources: Public Health Problem from Drinking Water Small town west of Springfield had 98 cases of gastrointestinal illness in June, 1990 Analysis of water reservoir showed possible blue-green algal bloom; call for survey came late in bloom Analysis of drinking water in hospital showed numerous algae including possible toxin formers.

3. My Professional Involvement as Teacher                                                                      BIO 530- Phycology Spring Semester, 2001 Thanks for joining this site at 02/22/2002 8:55:55 AM! (CST) Southwest Missouri State University Department of Biology Instructor: Russell G. Rhodes Office: Room 306, Kings Street Annex Phone: 836-6887 E-mail: rgr592f@smsu.edu The illustration to the right is a colony of Volvox, a member of the order Volvocales, Class Chlorophyceae.

Number of Occurrences of a Harmful Algal Blooms (Red and Brown Tides) Over the Past Decade http://www.redtide.whoi.edu/hab/HABdistribution/HABmap.html#U.S.%20Decadal%20Maps

Changes in Incidence of Marine Algal Toxin Occurrences http://www

Changes in Incidence of Marine Algal Toxin Occurrences 25 Years Later http://www.redtide.whoi.edu/hab/HABdistribution/HABmap.html#U.S.%20Decadal%20Maps

Headlines on HABs from Marine Habitats “Sea lion deaths linked to toxic algae bloom” (AP) –” The deaths of more than 400 California sea lions in 1998 have been traced to toxic algae that literally caused their brain cells to explode.” Source: http://www.canoe.ca/AllAboutCanoesNewsJan00/000105_alg.html “UC SANTA CRUZ RESEARCHERS TRACE TOXINS FROM ALGAL BLOOMS THROUGH THE MARINE FOOD WEB IN MONTEREY BAY” Source: http://www.ucsc.edu/news_events/press_releases/archive/00-01/01-01/toxins.html “CONSUMERS ADVISED NOT TO EAT SPORT-HARVESTED SHELLFISH, CRAB, SARDINES AND ANCHOVIES FROM MONTEREY BAY “ Source: http://www.wa.gov/wdfw/fish/shelfish/razorclm/domacid.htm

Moving to Freshwater Habitats and Things that Grab Your Attention “Thousands of Fish Are Dying in Texas Lakes” (WFAA.com, Dallas/Forth Worth) “Ingestion of the toxic material was implicated in the death of a pet dog and the illness of children who swam in the lake.” (Bernadette Rae Kenworthy, 2000. Master’s Thesis, U. Washington) Alligator killer- Cylindrospermopsis, a blue-green alga (Visual Contrast Sensitivity Test Center)

Other Things that Grab Your Attention in Freshwater Habitats Lesser Flamingoes Deaths in Lake Bogoria (Kenya Wildlife Service) Liver Failure with Death after Exposure to Microcystins at a Hemodialysis Center in Brazil. Jochimsen et al. (Brazil and CDC) NEJM 338:873-878, 1998. Water with toxic bluegreen algae suspected in high rate of liver cancer at locations in China Ueno, et al. 1996. Detection of microcystins ….inChina ….Carcinogenesis 17:1317-1321 http://biology.wright.edu/faculty/carmichael/research-r.htm

No Swimming due to Toxic Algal Bloom of Blue-Green Algae With permission, Chris Foxall, U. East Angilia, UK http://www.jei.uea.ac.uk/projects/waste_4.htm

Appearance of Blue-green Algal Bloom in Minnesota lake (with permission of Will Munson and the Minnesota Pollution Control Agency http://www.pca.state.mn.us/water/clmp-toxicalgae.html

And Closer to Home The algal blooms (Aphanizomenon) were responsible for early morning oxygen sags, which in turn led to partial kills of the largemouth bass and bluegill populations. http://www.epa.gov/OWOW/lakes/clnlake.html “539 (80 percent of water treatment facility samples that were submitted for testing) were positive for microcystins when tested by immunoassay.” http://www.awwarf.com/exsums/256.htm” “Microcystis, a blue-green algae that is harmful to humans and deadly to plants and fish, has returned to a small area of western Lake Erie after a 10-year absence. http://www.enn.com/enn-news-archive/1999/06/060399/microcystis_3534.asp

Countries- 42 (Worldwide) Canada- 6 provinces United States- 32 states Incidences of Toxic Cyanobacteria and Microcystin in Freshwater (compiled by Kotak, 1991; Yoo et al., 1995; Johnson, unpublished, Carmichael, AWWA Research News, Project No. 256) Countries- 42 (Worldwide) Canada- 6 provinces United States- 32 states 80% of samples from utility waters in US contained microcystin, a hepatotoxin

Understanding the Algae Basic Colors (Natural Groups) Golden Browns Blue-greens Diatoms Dinoflagellates

Brief Synopsis of Blue-green Algae Blue green due to phycocyanin and chlorophyll Forms can be unicellular, colonial, or filamentous Cells without nucleus, but can serve as spores and/or fix nitrogen and have gas vesicles

Known and Suspected Toxic Blue-Green Algae Anacystis Anabaena circinalis Anabaena spp. Anabaenopsis Aphanizomenon sp. Nodularia spumigena Cylindrospermopsis raciborskii Hapalosiphon Lyngbya Microcystis spp. Nodularia Nostoc Phormidium Planktothrix Schizothrix Trichdesmium Umezakia

Brief Synopsis of Golden Brown Algae Most harmful forms are flagellates Forms may be unicellular or colonial Many are covered with calcareous scales Golden brown due to xanthophylls and chlorophyll Some are very small, 2-5 microns in diameter

Known and Suspected Toxic Golden Brown Algae Aureococcus anophagefferens Pyrmnesium parvum Aureoumbra lagunensis

Brief Synopsis of Dinoflagellates Most are unicellular, few colonial About half are heterotrophic, others are photosynthetic Pigments are peridinin, other xanthophylls and chlorophyll Most have lateral “groove” Most are flagellate Most have eyespot                         http://www.bigelow.org/hab/cause.html                        http://redtide.net/cp.htm http://www.mote.org/%7Epederson/gbreve3.jpg and Phytoplankton Ecology Program at North Carolina State University. We borrowed the image from them courtesy of Dr.Dan Kamykowski.

Known and Suspected Toxic Dinoflagellates Pyrodinium bahamense Pfiesteria piscicida P. shumwayae Prorocentrum minimum Alexandrium (Gonyaulax) tamarense A. catenella Amphidinium carterae Dinophysis acuta D. acuminata D. norvegica Cochlodinium polydridoides Gambierdiscus toxicus Gyrodinium estuariale Gymnodinium Karenia brevisulcata Karenia brevis (Gymnodinium)

Brief Synopsis of Diatoms House of two pieces of overlapping glass Pores, slots, openings through frustule allow gases, nutrients, exudates to move Pigments are fucoxanthin and chlorophyll Forms are unicellular, colonial, or filamentous Some are motile by use of material movement in slot(s)

Known and Suspected Harmful and/or Toxic Diatoms Chaetoceros convolutus C. concavicornis C. danica Pseudo-nitzschia australis P. multiseries P. pungens

Brief Synopsis of the Raphiophytes Unicellular with no walls Biflagellate with apical insertion Large, 50-100 u, flattened Photosynthetic with chlorophyll, b-carotene and fucoxanthin Trichocysts Both freshwater and marine Chatonella sp. With permission of Mats Kuylenstierna http://www.marbot.gu.se/sss/others/Chattonella_sp.htm

Known and Suspected Toxic Raphidiophyte Chattonella veruculosa http://www.dnr.state.md.us/bay/hab/chattonella.html Chattonella sp. “Great amount of human made nitrate from the south part of the North Sea has probably stimulated the algae bloom.” Bloom responsible for fish kills in fish farms. http://www.utas.edu.au/docs/plant_science/HAB2000/poster_abstracts/docs/Aure_Jan.html Heterosigma akashiwo Fibrocapsa japonica http://www.utas.edu.au/docs/plant_science/HAB2000/abstracts/docs/Tyrrell_John_V.html

Acryonyms Used in Conjunction with Algal Toxins HABs- Harmful Algal Blooms NSP- Neurotoxic Shellfish Poisoning PSP- Paralytic Shellfish Poisoning ASP- Amnesic Shellfish Poisoning CTX- Ciguatera Toxins HWP- Hepatotoxic Water Poisoning PWP- Paralytic Water Poisoning

Understanding the Main Toxins of Algae Involved in HABs Neurotoxins: anatoxin-a, saxitoxins Hepatotoxins: cylindrospermopsin, microcystins, nodularins

Some of the Blue-green Algal Genera and Their Toxins (Not all species in a genus produce toxins.) http://lurac.latrobe.edu.au/~botbml/cyanotox.html http://lurac.latrobe.edu.au/~botbml/cyanos.html Algal Genus Toxins Anabaena Anabaenopsis Aphanizomenon Cylindrospermopsis Lyngbya Microsystis Nodularia Phormidium Planktothrix Anatoxins, Microcystins Microsystins Saxitoxins Cylindrospermopsins Aplysiatoxins Microcystins Nodularin Anatoxin Microcystins, Saxitoxins

Cyanobacterial HABs and Human Health Blue-green Microcystis and human health http://www.science.org.au/nova/017/017box03.htm Toxic Cyanoprokaryotes in resource waters: monitoring their occurrence and toxin detection http://www.eawag.ch/publications_e/proceedings/oecd/proceedings/Bouaich.pdf

Some Dinoflagellates and Their Toxins Gymnodinium catenatum- PSP (Mackenzie and Beauchamp, http://www.cawthron.org.nz/Assets/Gcat.pdf Alexandrium- PSP Pyrodinium- PSP Dinophysis- DSP Prorocentrum- DSP Gambierdiscus- CTX

Extent of Most Widespread Algal Toxin, PSP * With permission of Dr. F. M. Van Dolah http://www.chbr.noaa.gov/CoastalResearch/DiversityEssay.htm

Incidence of ASP in North America* *With permission of Dr. F. M. Van Dolah http://www.chbr.noaa.gov/CoastalResearch/DiversityEssay.htm

Limited Occurrence of Dinoflagellate Neurotoxin (NSP) due to Karenia (Gymnodinium) breve* *With permission of Dr. F. M. Van Dolah http://www.chbr.noaa.gov/CoastalResearch/DiversityEssay.htm

Incidence of DSP* *With permission of Dr. F. M. Van Dolah http://www.chbr.noaa.gov/CoastalResearch/DiversityEssay.htm

Incidence of Ciguatera toxin* *With permission of Dr. F. M. Van Dolah http://www.chbr.noaa.gov/CoastalResearch/DiversityEssay.htm

HABs and Bird Health Source: Field Manual of Wildlife Diseases, General Field Procedures- http://www.nwhc.usgs.gov Domoic acid- death in brown pelicans, Brandt’s cormorants Brevetoxin- suspected in death of lesser scaup Saxatoxin- suspected in death of common terns, herring gulls, Pacific loons, and others “Toxicosis”- suspected in free ranging ducks, geese, eared grebes, gulls, songbirds

Some Algal Toxins and Their Effects Nervous system- anatoxins brevetoxin, domoic acid, saxatoxin Liver damage- nodularin, microcystins, aphanotoxins, cylindrospermopsin Necroses- unnamed “bioactive substance” from Pfiesteria Suffication from gill clogging Cell surface interaction preventing egg development

Coincidence and Correlation “Over the past three decades, the frequency and global distribution of toxic algal incidents appear to have increased, and human intoxications from novel algal sources have occurred. This increase is of particular concern, since it parallels recent evidence of large-scale ecologic disturbances that coincide with trends in global warming.” Marine Algal Toxins: Origins, Health Effects, and Their Increased Occurrence by Frances M. Van Dolah Marine Biotoxins Program, NOAA National Ocean Service, Center for Coastal Environmental Health and Biomolecular Research, Charleston, South Carolina USA

Concentrations of N and P in Rural Runoff in the Midwestern U. S Concentrations of N and P in Rural Runoff in the Midwestern U.S. from Novotny and Olem (1994) Total N (mg/l) Total P (mg/l) Ratio Background levels 0.05-0.5 0.01-0.2 5:1 Cropland 9 1.2 7.5:1 Feedlots 920-2100 290-380 4-6:1 Grazed pasture 4.5 7 0.6:1

Trends in Eutrophication of Aquatic Systems Novotny, V. and H. Olem Trends in Eutrophication of Aquatic Systems Novotny, V. and H. Olem. 1994. Water Quality Prevention, Identification, and Management of Diffuse Pollution. VanNostrand Reinhold

Trends in Eutrophication of Aquatic Systems Novotny, V. and H. Olem Trends in Eutrophication of Aquatic Systems Novotny, V. and H. Olem. 1994. Water Quality Prevention, Identification, and Management of Diffuse Pollution. VanNostrand Reinhold

Causative Factors in Bloom Formations: Elser (1999) in “Freshwater Biology” Excess phosphorus (P) General eutrophic conditions Low N:P ratio Favorable light (bright), water conditions (calm) Daphnia (a zooplankter) dominance (there are exceptions) reducing competitors

N:P Ratios and their Significance N:P ratios less than 10:1 have been shown to favor dominance of heterocystous blue-green algae Significance of heterocyst in blue-greens as site of Nitrogen fixation

Biological Conditions that may Increase HABs Invasion of exotic animals Zebra mussels in Lake Erie Daphnia lumholtzi Shift in zooplankton grazers Algal products may reduce feeding by grazers http://aquaticpath.umd.edu/toxalg/btb.html Ability to survive enhanced UVBR by Alexandrium and Aureoumbra http://safari.gu.se/pages/925.html

Other Causative Factors Fertilizer runoff Livestock wastes Bright sun Calm waters Change in zooplanktonic grazers (Buskey, et al. 1997)

Preventative Measures for Toxins and Toxic Forming Algae Toxin- granulate activated charcoal (GAC) Toxin- powered activated charcoal (PAC) Algae- CuSO4, KMnO4 Above from Karner, et al (2001) in JAWWA Algae- Virus application http://www.sb-roscoff.fr/Phyto/Presse/Nature_Feb02.pdf

With permission of http://museum.gov.ns.ca/poison/redtide.htm Dinophysis acuta http://www4.fimr.fi/algaline/sheets/sheetman/dinoaut.htm With permission of http://museum.gov.ns.ca/poison/redtide.htm Prorocentrum minimum http://www.dnr.state.md.us/bay /monitoring/phyto/prorocentrum.html

                                                                                                   Gymnodinum catenatum, colony and resting cyst With permission of F. Hoe Chang http://www.niwa.cri.nz/pubs/bu/02/blooms.htm

Problems in Missouri Lack of equipment to routinely do algal assessment in water treatment facilties Insufficient time allocated for water monitoring by treatment plant operators In some locations failure to recognize the interests and abilities of treatment plant operators to provide proper monitoring as they see the need

Solutions in Missouri and Elsewhere: Enhanced Monitoring at a Basic Level Distribution of surplus microscopes to water treatment facilities lacking such equipment Provision of Secchi disks or other means to monitor turbidity of water prior to entry into plant Periodic algal assessment of domestic surface water supplies throughout Missouri on a priority basis Early treatment of reservoir to reduce potential for algal blooms

Water Collection- Biomass Device in center is a Kemmerer bottle. It is lowered to a specific depth and a “messenger” is sent down the cord to trip the traps.

Counting Algae: Bright Field and Phase Compound Microscope with TV and monitor

Counting Chambers Top view is a haemocytometer counting chamber (small algae) Bottom view is Sedgwick-Rafter counting chamber (big algae)

Inverted Microscope and Identification Manual: Utermohl Technique using Settling Chambers.

Utermohl Technique for Counting Algae (Quantitative)

Chemical Assessment Techniques for Monitoring Toxic Algae and Algal Toxins Environmental Sampling Processor (ESP) for identifying toxic eukaryotic algae: http://www.mbari.org/news/news_releases/2001/jun05_ESP.html Enzyme linked immunosorbent assay (ELISA) for microcystin recognition: http://www.nwhc.usgs.gov/ Neuroreceptor assays for saxatoxin http://www.nwhc.usgs.gov/ High Throughput Receptor Binding Assays For Phycotoxins

Enzymy Linked Immunosorbent Assay (ELISA) Technique An excellent overview of the ELISA Technique: http://www.hhmi.org/grants/lectures/1996/vlab/

Links http://courses.smsu.edu/rgr592f http://courses.smsu.edu/rgr592f/courses/bio530.htm http://biology.smsu.edu/phycology/toxic-algae.htm http://biology.smsu.edu/phycology\Algal Identification and Survey Class\class\Default.htm http://courses.smsu.edu/rgr592f/algae_in_domestic_water_supplies.htm Definitive Blue-green algae toxin information http://www.who.int/water_sanitation_health/Recreational_water/Recreawat-II.pdf http://www-cyanosite.bio.purdue.edu/cyanotox/cyanotox.html The IOC Harmful Algal Bloom Programme

Resources Novotny, V. and H. Olem. 1994. Water Quality Prevention, Identification, and Management of Diffuse Pollution. Van Nostrand and Reibold. 1054 pp. Smith, V. H. 2001. Blue-green Algae in Eutrophic Fresh Waters. Lakeline- 2001:34-37. Anderson, D. M. and D. J. Garrison, eds. 1997. The ecology and oceanography of harmful algal blooms. Limn. Ocean. 42 (5 part 2): 1009-1305. Harmful Algal Blooms, Ninth Conference, Tasmania, 2000.