### APPENDIX IV: MEETING AGENDA AND WORKING GROUP ASSIGNMENTS

#### November 13, Tuesday

Afternoon/Evening

Out-of-town participants arrive at MIA, take cab or shuttle from baggage claim area (lower concourse) to Sheraton Royal Biscayne (20-30 min drive and cost will be about $25 for a single cab). Enclosed is an annotated list of restaurants on Key Biscayne near the hotel if the hotel restaurants don't appeal to your tastes. We'll leave a message at the desk if there is any social "event" planned; most likely we'll just congregate in the hotel bar.

#### November 14, Wednesday

0800 Depart Hotel Lobby (We will have a rented multi-passenger van and personal cars to take you the 5 minute drive to RSMAS)

0830 RSMAS Library Chart Room
Opening comments by:

- Bruce Rosendahl, RSMAS Dean
- Don Olson, Member GLOBEC Steering Committee
- Dan Morse, Editor, Biotech/Ocean Sci. Initiative
- Lew Incze and Pat Walsh, Workshop Organizers

0930 Break into Working Groups I and II (see attached lists): Group I to Library Conference Room, Group II in Library Chart Room.

COFFEE AND DOUGHNUTS WILL BE BROUGHT IN AT MID-MORNING

1130 Break for Lunch at RSMAS Cafeteria (you'll need to pay your own way and keep receipt)

1300 Reconvene Working Groups I and II

1445 Break for Coffee at RSMAS Cafeteria (closes at 1515)

1515 Reassemble in Chart Room for a) plenary discussion of how groups are progressing; what problems they are encountering; what they see as things which will need addressing; discussion of group objectives and assignments for the following day; b) any further discussion that individual groups wish to have.

1730 Adjourn to RSMAS Bar for R and R. We will arrange for a group dinner at a local eatery (Cuban, Thai, etc.)-we hope all of you can make it. We'll coordinate transportation back to the hotel and then to dinner. We'll try to leave time for those who want to run or swim before dinner.

#### November 15, Thursday

0800 Depart Hotel Lobby

0830 General remarks, objectives and agenda for the day.

0900 Split into Working Groups III and IV (see lists). Note: Day 2 working groups are tentative, with changes subject to Day 1 discussions.

COFFEE AND DOUGHNUTS WILL BE BROUGHT IN AT MID-MORNING

1130 Break for Lunch at RSMAS Cafeteria

1245 Reconvene Working Groups III and IV

1445 Break for Coffee at RSMAS Cafeteria (closes at 1515)

1515 Working Group Reports (ca. 15 mins ea.); plenary discussions; any further discussions that individual groups wish to have.

1700 Closing remarks; wrap-up details; writing/reviewing deadlines for workshop report; where do we go from here?

1730 Adjourn for socializing. For the time being no specific plans are made regarding a trip to the bar, dinner, etc. If people want, we can organize something; we'll bring this up on Thursday morning.

#### Overview and Objectives

Before we proceed to working group discussions, it is useful to consider the framework in which the deliberations of each group should take place.

GLOBEC wants, ultimately, to understand how marine populations respond to physical variability of the environment. Thus, we must have the ability to assess certain critical physiological parameters (rates and condition) with sufficient detail to resolve differences among animals on spatial and temporal scales typical of the ocean's variability. Concurrently, this will require a much better ability to sort animals, involving speed and accuracy and no change in critical proteins or molecules. Ideally, we would be able to sort samples and make physiological assessments rapidly enough and efficiently enough (in terms of space and personnel) that they both can be done at sea with reasonable turn-around times. Sorting probably would be based on specific genetic, biochemical or molecular markers and should be adaptable to increased levels of automation. In this way, the biotechnology eventually could become an interactive part of research cruises. This should provide many spin-off benefits to shore-based research efforts as well.

We do not expect to design final solutions to these goals as an outcome of this workshop; this would be a tall order under the best of circumstances. Rather, the workshop objective is to summarize results by drafting a "call for proposals" with supporting comments for each of the two goals, that is, development of techniques for 1) physiological measurements and 2) rapid sample sorting (and genetic identification). These drafts will reflect the expertise of this group of scientists, your deliberations here at this workshop, and your consensus of what is feasible, what is desirable, and what is the most progressive way to proceed in terms of research. GLOBEC is strongly committed to supporting the necessary biotechnology development and biotech/oceanographic collaborations and seeks the guidance of this group in developing its strategies. If our recommendations are accepted, biotechnology RFP's will be issued through the proper agencies in the near future, and we expect that many individuals from this group will be applying for portions of the available funds.

We have asked two individuals from each group, one each from the oceanography and the biochemistry/molecular biology "sides", to lead the discussions and prepare a brief report of results from each working group. The results should include highlights from the discussions and a draft of how you would want to see an RFP on the topic focused. Leaders are free to solicit additional help as they see fit for note-taking, report-writing, etc. The end product will be a report to the GLOBEC Steering Committee for their action.

#### Working Group I-Physiological Condition and Rates

José Torres & George Somero Co-Discussion Leaders
Charlie Miller | Gary Kleppel | Elizabeth Clarke
Roger Mann | Michelle Wood | Gail Theilacker
Steve Hand | Bruce Sidell |

The assessment of physiological condition, age and growth and developmental rates of individual zooplankters, and the variability of these parameters, is an important part of understanding the influences of physical processes on population dynamics. This working group is charged with identifying techniques, which are currently available or which show promise for rapid development, to assess the age, physiological rates and condition of zooplankters. In order to guide the discussion somewhat, we've posed several questions below that should be answered regarding the identified methods.

1. What biochemical/molecular techniques are available to assess physiological condition and rates in the following (or additional) categories?
   - general performance capabilities (e.g., locomotion);
   - morbidity/low physiological capability (e.g., point of no return);
   - egg production rates;
   - "diapause" state;
   - growth rates;
   - developmental stage;
   - age.
2. Are there particular taxa/species which should be targeted, either because the information gathered is most important to the underlying scientific question(s), or because the methods will be particularly adaptable?
3. Are the methods adaptable to individual zooplankters, or must individuals be pooled to obtain enough tissue? How many?
4. Are the methods adaptable to ship-board use? If so, how much time is required to obtain results? If not, what degree of sample sorting and mode of preservation (e.g., freezing, ethanol, etc.) are suitable or unsuitable?
5. What equipment needed? Specialized or off the shelf? How expensive? How time consuming is the process? Can the methods be mechanized or automated?
6. Is laboratory development of the method necessary, and if so, how much spin-up time is anticipated before field samples can be meaningfully analyzed?
7. How easy/difficult will it be to train technical personnel to use the method routinely?

#### Working Group II-Population Genetics and Genetic Markers

Michael Lynch & Dennis Hedgecock- Co-Discussion Leaders
Mark Huntley | Peter Ortner | Dan Morse
Jason Hofman | Diane Stoecker | Doug Crawford
Dave Hillis | Steven Fain | Kelly Thomas

An important part of understanding the influences of physical processes on population dynamics is knowing what species and what relative abundances of species are present at various points in time and space, and what physical factors influence gene flow. Traditional (e.g., morphological) methods have shown to be inadequate in resolving some particular species identification problems. Also, traditional batch preservation of plankton tows, with subsequent sorting and enumeration are tedious and time consuming. Typically many months transpire before biological oceanographers can match their data to the corresponding physical data that were collected and examined in real time. One ultimate, perhaps presently unrealistic, goal is to enable biologists to collect real time data on distribution, abundances, and species/genetic composition of zooplankters. The charge of this working group is to assess the applicability of recent developments in molecular biology and genetics to these problems and to determine the feasibility of achieving the above stated goals. In order to guide the discussion somewhat, we've posed several questions below that should be answered regarding the identified methods.

1. What biochemical/molecular techniques are available to define genetic structure in zooplankton populations, and to quantitatively assess species composition.

Questions 2-7 as for Group I

#### Working Group III-Feeding Rates and Dietary Components

Gary Kleppel & Steve Hand- Co-Discussion Leaders
Elizabeth Clarke | Gail Theilacker | Mark Huntley
Diane Stoecker | José Torres | Steven Fain
Dave Hillis | Kelly Thomas |

Feeding rates and composition of the diets of organisms are two variables that might change over moderate spatial scales or might be quickly altered by physical forcing. These, in turn, probably affect condition, growth, egg production and other important aspects of individual physiology. To understand linkages between physical forcing and population dynamics requires advances in our ability to assess feeding and diet. In order to guide the discussion somewhat, we've posed several questions below that should be answered regarding the identified methods. They follow the same themes used on Day 1.

1. What molecular/biochemical techniques would enable rapid and sensitive assessments of feeding rates? What about quantifying dietary composition?

Questions 2-7 as before.

#### Working Group IV-Sampling

Peter Ortner & Bruce Sidell- Co-Discussion Leaders |
Roger Mann | Michelle Wood | Michael Lynch
Dan Morse | Jason Hofman | George Somero
Doug Crawford | Dennis Hedgecock | Charlie Miller

It is imperative that the molecular/biochemical techniques that we have discussed in other groups be combined effectively with sampling methods. Sampling must be of a scale (e.g., depth resolution) and speed to satisfy field questions, must handle organisms in acceptable ways, and must deliver these organisms in a manner that can be handled by the shipboard (biotech) procedures. To resolve the necessary spatial and temporal scales of interest to GLOBEC, all of this will have to be done often, probably several days running at a time. New procedures may have to be invented to make the linkage feasible. There are individuals in this group who can make reasonable guesses about the demands of sampling to answer typical questions of interest in oceanic ecology; the charge to the group is to consider what it would take to put the biotechnology and field sampling needs together effectively. There are at least two distinct but related needs. First, rapid identification of the plankton community (or selected parts of it: Working Group II). Second, delivery of desired individuals (e.g., species, stage) to the apparatus of physiological assessment (Working Groups I and III). Questions that might be asked are:

1. How many depths might be sampled at a single station?
2. How quickly will samples be delivered to the surface; by what methods and in what quantities should they be collected?
3. How many replicates should be run from each depth/sample for a single species for a single parameter (e.g., enzyme system)? You should consider here, and elsewhere when appropriate, the different demands of a small meroplanktonic larva, such as a bivalve; a copepod or copepodid; and a larval fish.
4. What would the transfer time be like once the sample is on deck until it is being processed by the appropriate method? Is this a problem?
5. How much space and how many people would the necessary biotechnology set-ups require to keep pace with the desired sample delivery rates, especially if 3 different taxa were being investigated? Is this doable on a ship? Can this be improved?
6. Is significant automation feasible within the next several years? Measure its potential impact in terms of a) speed and b) space and personnel requirements (questions 4 and 5).
7. What taxa and techniques would you recommend be attempted first? Why? What would it take?