Leslie Sturmer & JUNE 5-6, 2019 Dr. Huiping Yang FWC Sen. Kirkpatrick Marine Lab UF/IFAS Molluscan Shellfish Program 11350 SW 153rd Court, Cedar Key, FL 32625 NOAA National Sea Grant (Presentations & Demonstrations) • Function of Microalgae • Mass-Culture Strategies • Nutrition of Bivalves • Chemical Ecology of Cultures • Phytoplankton as Bivalve Foods • Media preparation & Aseptic transfers • Quantification Methods www.nefsc.noaa.gov/nefsc/Milford MICROALGAL CULTURE WORSHOP June 5-6, 2019 FWC Senator Kirkpatrick Marine Lab 11350 SW 153rd Court, Cedar Key, FL Guest Speaker: Dr. Gary Wikfors Director, NOAA Fisheries Milford Laboratory, Milford, CT AGENDA Wednesday, June 5, 2019 10:30 - 11:00 AM Registration 11:00 - 11:30 AM Welcome and Introductions 11:30 - 12:30 PM Introduction to Microalgae 12:30 - 1:15 PM Lunch (on site) 1:15 - 2:15 PM Function of Microalgae 2:15 - 3:15 PM Mass-Culture Strategies 3:15 - 3:30 PM Break 3:30 - 4:30 PM Nutrition of Bivalves 4:30 - 5:00 PM Discussion Thursday, June 6, 2019 8:00 - 10:00 AM Hands-on Demonstrations: Quantification methods, Aseptic transfers, Media preparation Meet at UF/IFAS Nature Coast Biological Station, 552 1st St, Cedar Key, FL 10:00 - 10:30 AM Break (head back to FWC Marine Lab) 10:30 - 11:30 PM Chemical Ecology of Microalgal Cultures 11:30 - 12:30 PM Natural Phytoplankton as Bivalve Foods 12:30 - 2:00 PM Box lunch, Q&As, Discussion and Wrap-up Dr. Gary H. Wikfors Chief, Aquaculture Sustainability Branch / Lab Director Email: [email protected] Education •Ph.D. Phycology - University of Connecticut •M.S. Biology - University of Bridgeport CT •B.S. Biology - University of Maine at Orono Gary’s terminal degree is in Phycology – the study of algae – but he always has worked at the intersection of phytoplankton and the bivalve mollusks -- such as oysters, clams, scallops, and mussels -- that derive their nutrition from phytoplankton. Gary has studied trophic transfer of pollutants from phytoplankton to bivalves, biochemical nutrition of shellfish, and harmful- algal effects upon bivalves. Much of his research has employed a laboratory- based, experimental approach, but he also has been involved in large, multidisciplinary field studies. Gary was an early adopter of flow-cytometry for microalgal applications, and use of this technology has sparked a subsequent interest in cellular immunity in bivalves and other invertebrates. As Chief of the Biotechnology Branch, Gary has a hands-on role in several current team initiatives: 1) Nutrient bioextraction using shellfish aquaculture, 2) Probiotic bacteria for use in shellfish hatcheries, and 3) Shellfish cellular immune response to environmental variation. ENCYCLOPEDIA OF AQUACULTURE 520 MICROALGAL CULTURE MICROALGAL CULTURE Table 1. List of Currently Recognized ‘‘Microalgal’’ Classes and Representative Genera Used in Aquaculture GARY H. WIKFORS Representative Northeast Fisheries Science Center Genera Used in Milford, Connecticut Class Common Name Aquaculture Cyanophyceae Blue-green algae Spirulina OUTLINE Prochlorophyceae Prochlorophytes None Rhodophyceae Red algae Porphyridium What are Microalgae? Prasinophyceae Scaled green algae Tetraselmis Pyramimonas Why Culture Microalgae? Chlorophyceae Green algae Chlorella Microalgae for Extractable Chemicals Dunaliella Microalgae as Aquaculture Feeds Haematococcus Cryptophyceae None Cryptomonas How do Microalgae Work? Rhodomonas Energy Chlorarachniophyceae None None Materials, or Nutrients Euglenophyceae None None Microalgal Growth D Population Growth Dinophyceae Dinoflagellates Crypthecodinium (Pyrrophyceae) How are Microalgae Cultured? Chrysophyceae Golden-brown algae None Containers Raphidophyceae None None Energy Eustigmatophyceae None Nannochloropsis Xanthophyceae None None Materials (Tribophyceae) Culture Management Bacillariophyceae Diatoms Thalassiosira What Innovations are Expected? Chaetoceros Nitzschia Bibliography Dictyophyceae None None Pelagophyceae None None Haptophyceae None Isochrysis WHAT ARE MICROALGAE? (Prymnesiophyceae) Pavlova By direct translation from Latin, microalgae are ‘‘little seaweeds.’’ However, defining microalgae further is not simple, because the microalgae represent a the category is necessary (see Table 1). Higher level sys- taxonomically diverse group of organisms, rather than tematics of these groups are under revision; hence, only a single, phylogenetic category. A functional definition the class level is specified. The diversity of the group is of microalgae might be ‘‘photosynthetic single-celled or underscored by the presence of both prokaryotic (not con- colonial microorganisms’’; however, most of these microbes taining a nucleus, Cyanophyceae, or cyanobacteria) and are able to grow without light if dissolved sugars eukaryotic (nucleated) taxa. The eukaryotic groups are are provided. Microalgal cells range in size from one thought to have arisen from the incorporation of photo- micrometer — roughly the size of a bacterium — to several synthetic prokaryotes (or, subsequently, photosynthetic hundred micrometers (1 µm) — barely visible to the naked eukaryotes) within protozoan-like host organisms. This eye. Colonies and chains of some microalgal cells can hypothesized process is referred to as the endosymbiosis attain a length of several centimeters 2.5cmD 1in..This theory. Evidence from both electron microscope studies of group of organisms contains remarkable morphological microalgal cells (usually focused upon numbers and types diversity, with shapes ranging from simple spheres to of membranes within the cell) and more recent molecular the ornate, silica shells of one group, the diatoms. sequencing work indicates that endosymbiotic creation of Many microalgae are motile, propelling themselves with ‘‘new’’ organisms may have occurred a number of times in flagella, by amoeboid motion, or by gliding on extruded evolutionary history, leading to the diversity in morphol- mucilage. Microalgae are found in an astonishing range of ogy and physiology seen today. This diversity, especially in habitats, such as in fresh, saline and hypersaline waters, terms of physiology, provides opportunities for the current in polar ice, in soil, attached to plants and animals, and potential use of these organisms in the aquaculture and even in symbiotic relationships with fungi (e.g., industry. lichens) and animals (e.g., corals). Historically, many of these organisms have been claimed and named by both zoologists and botanists; therefore, taxonomy has WHY CULTURE MICROALGAE? been, and remains, problematic. From the perspective of aquaculture, there are common characteristics that There are two main reasons for which microalgae are warrant their consideration as a functional group; the grown: (1) for extractable chemicals and (2) as feeds for foregoing definition will suffice for this discussion. aquacultured animals. Efforts to produce foods for direct As ‘‘microalgae’’ is a functional rather than phyloge- human consumption have met with limited success, and netic group, a list of taxa that would reasonably fit within crops such as the cyanobacterium Spirulina remain in the MICROALGAL CULTURE 521 realm of ‘‘dietary supplement,’’ rather than food crop; these such as temperature and salinity. An alga’s ability to cases are considered along with extractable chemicals. coexist with or exclude microbial contaminants, ranging from bacteria to fungi and protozoans, also is critically Microalgae for Extractable Chemicals important in most commercial applications. A number of specific, clonal strains of microalgae have Chemical analyses of microalgae have led to the discovery been found, empirically, to possess desired characteristics of many novel chemical compounds, some of which are and are in wide use; these can be obtained from useful as food additives, pharmaceuticals, cosmetics, or aquaculture supply companies, academic institutions, and in other high-value applications. Examples of microalgae government-funded institutions that maintain microalgal currently commercially cultured for extractable chemicals culture collections. The strain level of identity is include (1) Dunaliella for ˇ-carotene, a human nutritional appropriate, because microalgal taxonomy remains in a supplement; (2) Haematococcus for the pigments astax- state of continuing development and because different anthin and canthaxanthin, which are used as coloring isolates of the same taxonomic species may differ widely agents in salmon feeds; (3) Crypthecodinium for docosa- in growth or nutritional characteristics relevant to their hexaenoic acid (DHA), which is incorporated into infant use as aquaculture feeds. formula; and (4) the aforementioned Spirulina,whichis used as a dietary supplement or additive to cosmetic prod- ucts. In most cases, the chemical compound of commercial HOW DO MICROALGAE WORK? interest is present as a small fraction of the total algal biomass; therefore, the scale of cultures for commercial Energy production generally is in the range of many cubic meters 3 Microalgae are referred to as autotrophs, a word that 1m3 D 35.3ft . When a selected portion of the algal translates literally as self-feeding. This term is wholly biomass is extracted and refined, the presence of chemical appropriate because the process of photosynthesis, by and biological contaminants within production cultures is which light energy is converted to chemical energy, not relevant, unless overall production is constrained or an provides sugars that are subsequently eaten, or burned, undesired chemical is coextracted with