Presentation to ABO 7th Annual Algae Biomass Summit, Orlando, October 2, 2013.

Willie Wilson, Ph.D.

Director The Provasoli-Guillard National Center for

Marine Algae and Microbiota (NCMA)

Bigelow Laboratory for Ocean Sciences

Culturing Microalgae at the Small Scale

Culturing Microalgae at the Small Scale

A Case of Perpetual Care. Why cultivate (domesticate) algae?

What are your objectives?

Does the need justify the effort?

Are there other means of doing what you need?

What facilities are needed?

A defined area of controlled temperature to grow cultures.

Facilities for making up media.

Means of sterilizing the media?

What type of media do you need?

Simplest to the most complex!

Temperature/light regulated growth chamber

Laboratory North Facing Window

The major control is temparature.

Light intensity and duration is next.

Alarms or warning controls.

Maintenance in tube culture –

Small batch mode

30-40 mL of culture

Maintained with frequent transfers – ideal is monthly intervals

Needs:

Sterile glass culture tubes

Sterile medium

Appropriate growing space

Handling materials – glass pipettes heat or autoclaved.

Laminar flow hood

Bacteriological sterile technique needed!

New curators if the collection becomes of any size.

Media making do’s and dont’s

Defined media - can be made in specified limits the same each time

many species will not grow as well as in other media

costly since base water is usually highly purified

excellent for physiology or where conditions need to be defined within narrow limits

Examples: Wilson’s NH15 and Aquil

Enriched sea water media (ESW)

requires good quality, off shore seawater. Great if you live near the Gulf of Mexico, Sargasso Sea or Eastern Mediterranean

Besides having a mode of acquiring the sea water (ships, pumping systems, cold dark storage) you must be aware of seasonal variations.

Example of successful sea water enriched media.

Oldest and a stabile stand by – Erdschribers Media

Problem – soil extract plus N and P

Advantage – grows nearly everything (Ask Richard and Maria)

The Provasoli variations –

Sea water enrich with trace metals tailored to the species

Precipitation often a problem

The Guillard Series – beginning with f/2, K medium, L1 medium et al.

With understanding the need for selenium, many of the problem species have been cultivated. Open ocean species are no longer a problem.

Work stations.

Think about your culture space, walk-in incubators are convenient, and flexible.

Reach-in incubators work well. Note: light level variability. Different culture vessels. Ways to label growth medium.

Accurate labeling is paramount.

Seaweeds are algae too! ..and they are easy to culture.

Cryopreservation save collection managers a small fortune…..

A role for bioresource centers (BRCs) in the 21st century.

• A Bioresource Center • Mission and Vision • Microbial (Algal) Commodities • An NCMA Snapshot • Leading an Algal Industry

Cluster in Maine Practicing the ART OF POSSIBILITY

Time to move???

NCMA’s status is a service center of Bigelow Laboratory for Ocean Sciences, a non-profit 501(c)(3) organization. Income derived from federal grants, culture fees and services support the functions of our mission. NCMA’s mission is to serve as a central bioresource center whose core activity is to receive, maintain and distribute living cultures of marine and freshwater algae, bacteria and viruses; to provide technical expertise and services; and to provide educational resources for culture isolation and curation to scientists, educators, biomedical researchers, and businesses worldwide. NCMA’s vision is to become the global leader in the provision of algae, protozoa, bacteria and viruses, together with related products for academic research and commercial ventures. By 2020 the NCMA brand will be widely recognized for first class customer service and intellectual capability.

Microbial ‘Commodities’ Oceanic microbiota

(protists, algae,bacteria, archeae, viruses) are the

largest untapped biotechnological resource

on the planet.

Glynn Gorick

The NCMA holds representatives from 39 Classes of Algae (all the major photosynthetic groups)

Apicomplexa Aurearenophyceae Bacillariophyceae Bicosoecophyceae

Ciliatea Charophyceae Chlorarachniophyceae

Chlorophyceae Chromerida

Chromophyte

Chrysomerophyceae Chrysophyceae

Bolidophyceae

Coscinodiscophyceae

Cryptophyceae

Cyanophyceae

Dictyochophyceae

Dinophyceae

Euglenophyceae

Eustigmatophyceae Fragilariophyceae

Glaucophyceae Pelagophyceae

Phaeophyceae

Phaeothamniophyceae Pinguiophyceae

Porphyridiophyceae

Prasinophyceae

Prymnesiophyceae

Raphidophyceae

Rhodophyceae

Schizocladiophyceae Synchromophyceae

Synurophyceae Trebouxiophyceae

Ulvophyceae undetermined

Apicomplexa

Aurearenophyceae

Bacillariophyceae

Bicosoecophyceae

Bolidophyceae

Charophyceae

Chlorarachniophyceae

Chlorophyceae

Chromerida

Chromophyte

Chrysomerophyceae

Chrysophyceae

Ciliatea

Coscinodiscophyceae

Cryptophyceae

Cyanophyceae

Dictyochophyceae

Dinophyceae

Euglenophyceae

Eustigmatophyceae

Geographical Location of Strains

NCMA: A Snapshot

• The world’s largest and most diverse living archive of marine microalgae.

• 35 years in the algae business. • Started as a algal seed stock for the aquaculture industry. • A repository for public and private collections of algae. • 2,720 strains of marine, brackish, hypersaline and freshwater

algae; including cyanobacteria and macroalgae. • 359 genera and 723 species. • 6 products available for every strain. • 5 growth temperatures (polar to tropical). • On-site & off-site back-up and cryopreservation. • Expansion to include new Bacteria and Virus collections. • New web site to enhance customer experience (Oct 2013).

Products and Services

• Starter cultures • Nucleic acids from algae • Culturing Techniques Courses • Private collections • International Depository Authority (patent depository) • got algae? T-shirts! • Research services • Isolations/clean up/taxonomic ID • Services through our partners (e.g. powdered algae) • Other service centers at Bigelow

– Single Cell Genomics – Flow Cytometry – Analytical Services – Seawater Facility

VISION To create a complete algal supply chain; from seed cultures to biomass infrastructure, processing, testing, and development of market opportunities; that will allow us to lay a foundation to stimulate algal economic development, and make Maine one of the top states in the U.S. to run an algal-based business. It will be a process driven by innovation, education, collaboration, and branding.

Proposal for an Algal Industry Cluster in Maine

NCMA Entrepreneurship The Future is Bright!

Because the oceans of the world represent a vast frontier of undiscovered compounds and organisms, the prospect exists for the NCMA to work with numerous private industries to explore commercial opportunities for new products and technologies in a variety of commercial markets.

Innovation Gets You Recognized

The NCMA Team

Questions

Dr. Willie Wilson

wwilson@bigelow.org https//ncma.bigelow.org

207-315-2567 (x310)