The Risinq and Fallins of Chaetoceros

Nyles Toquehi

September 3,1999 MarBec Summer

Vnderqraduate Research Proqram

Table Of Contents

Abstract

Introduction

Methods and Mater:a!

Continue

F2 / Herinq Solution

Nusalts So!ution

Results

Fiqure #I

Figure #2

Continue results

~:scuss!on

Sooclone fractionator

Evaluation of leorninq

Continue

Four Staqes of Chaetoceros

Acknow!edqments

PP.GE 2

PAGE 3

PAGE 4

PAGF 5-

PAGE 6

PAGE 7

PAGE 8

PAGE 9

PAGE 10

PAGE I!

PAGE 12

PAGE 13

PAGE 14

PAGE 15

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PASE 2

Abstract

With my pro.iect, I want t o develop ways t o improve the way of harvestinq

Ghaetoceros (Phytoplankton). If harvesting of Chaetoceros can be improved, i t can be

applied in the feedinq of different types of zooplankton (artemia, rotifers, and copepods).

Zooplankton can be fed t o different types cf larval finfish and some types o f mol!uscs.

The conventional foam fractionator has been found t o be rouqhlv 23% efficient.

For people who use these cells for feedinq and other purposes, th is means more than four

times the work.

Unfortunately, the fractionator tha t 'I desiqned - failed, but I am sti i l workinq - on

making this desiqn work. Hopefully, a few modifications t o the desiqn will allow the water

t o circulate faster with smaller bubble size. This is currently beinq done with a new

venture system. The results of this revision will be reported t o MarBec's internship

proqram . - leaders.

Irrfradlactian:

The harvestins - of Choe?oce.ros can be useful in several different applications. . .

Chae?oceroscan be used for rearins shrimp larva, feeding zooplankton (co~epads, artemea,

and ratifers). and for ~harmaceutical purposes. Beinq able to concentrate ce!!s into a

sma!!er container can improve water quality by reducinq the chances of contamination and

feed e f f iciencv. With my nro.lect. 1 hope to be able to improve the way of harvestinq -

Gaetocerm and perhaps other . phytoplankton. .

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Heri nq Fert i l i zer

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Na,EDrA FeCI.6H-0 CuSo4SH20 ZnSOa7H70 CoCl,hU20 Ma Mo042H-O

Biotin Cyanocobaiamin Thiamin HCI inerts

One qram Nutrients

Nuselt Fertilizer

Items

NaNO. NaH PO..H-0

Na,EDTA FeCI-6 H70

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One qram Nutrients

?AGE 5

The f i rs t par t of my pro.iect dealt determininq with the efficiency of a

conventional foam fractionator and comparina tha t t o nonfractionated contro!. This was

done by culturinq - two tanks t o equal cell densities. Then, one tank is centrifuqed and the

other tank is foam fractionated and then centrifuqed. - The paste is taken t o the Bio-

system Enqineerinq - - lab a t the Universiiv of Hawaii t o be freeze dried.

The second par t of my pro-ject was t o desiqn and build a fractionator of my own. I

desiqned - a fractionator out of acrylic usinq - a Seaclone protein skimmer as my model.

l%e last part of my . . pro-iect - was t o t es t the efficiency of my fractionator. First,

two tanks were cultured t o equal cell densities. Then, one tank was centrifuaed and the

other tank was foam fractionated and then centrifuqed. The paste was individually

weiqhed and then taken t o the Bio-system Enqineering lab a t the University of Hawaii t o

be freeze dried.

Results and Discussiar.

The first experiment of my pro.iect was to measure the efficiency of the

conventional foam fractionators (Figure #lf. Two tanks were mixed, using a samp pump, t o

equal cell densities of 850,000 Lhaetocerosper liter. Then one tank was centrifuged

while the other tank was foam fractionated and then centrifuqed. The tank that was

centrifuqed - . produced, 84.4 qrams - of paste and 14.3 grams in dry weiqht. The tank that

was foam fractionated produced 2.5 qallons - of foam (concentrated fiaetoceros) to be

centrifuqed. The 2.5 qallons of foam produced 19.55 qrams of paste and 1.85 qrams in dry

weiqht. -

The second experiment was a duplication of the f i rs t experiment. The only

difference was the cell density which was 2,690,000 fiaetacerosper liter (Fiqure - #2).

The centrifuqed tank produced 169 grams of paste and 27.05 grams in dry weiqht. The

fractionated tank produced 5 qallons of foam [concentrated fiuefocet-0s) to be

centrifuged. P i s produced 92.85 qrams of paste and 9.35 qrams in dry weiqht. Between

the two experiments, I t showed that the foam fractionators were only 23% efficient in

removing cells from the water.

The second part of my project, was to build a fractionator based on a protein

skimmer called the Seaclone. The Seacione uses a Venturi system to get a cyclone effect

in a cylinder. The cyclone ef fect is able to concentrate cells into the center allowing the

air from the Venturi system to push the cells up to be collected and / or harvested.

PACF 11

The last part of my project was to find and compare the efficiency of the Seaclone

fractionator and the foam fractionator. The experiments were to culture two tanks to

equal cell densities and fractionate one tank and centrifuge the other.

The f i rst experiment had 530,000 Chaetocerosper liter. The centrifuged tank

produced 91.2 prams of paste while the fractionated tank was not able to produce more

than five liters (Too l i t t le t o centrifuge).

The second experiment had 3,150,000 Chaefoceros per liter. The centrif uqed tank

produced 183 qrams of paste while the fractionated tank was not able to produce more

than five liters !Too l i t t le t o centrifuge).

Biscussion

I%rouqh - my ~ experiments, I have found tha t the foam fractionators are not very

efficient, but through some other experiments done by Andrew Csorsa (Graduate Student

a t the University of Hawaii), micro-bubble air stones wiil make foam fractionation much

more efficient. For the future, some experiments should be conducted t o f ind out the

efficiency of different air stones for foam fractionators.

The fractionator tha t I designed may have failed it 's initial experiments but I st i l l

feel with a l i t t le more time and thought, I can modify and prove tha t this fractionator is

much more efficient in harvesting Chaefoceros.

The seaclone fractionator tha t I designed, did not work as well as expected! but

more than Iikeiv it was due t o my design. I have three theories on why the fractionator

did not work. There might not have been enough rotation of the water due t o create a

cyclone e f f ec t due t o not enough flow ra te of air and water pressure. Another possible

theory would he tha t the bubble size may have been too large t o collect any cells. The last

theory is tha t the fractionator might not have been wide enough t o cause the proper

cyclone e f f ec t (Figure #4)

Evaluation of learninq

Throughout this summer, I found that I was able to accomplish a lot of different

side experiments that might be useful to someone using an open system that is culturing

Ch~e ~ Q ~ ~ ~ P Q S .

One experiment that I was able to accomplish was to implement a schedule of

providing different nutrients for Chaetocerosfor optimal cellular growth. I found that

Chaetoceros has four different cycles. The f i rst is the beginninq - stage. This stage is

when the cu!ture needs to establish itself in the tank. The second cycle is the growth

staqe. This is when the cells begin to become well established in the tank by asexual

reproduction. The Third staqe is the stabilized stage. This stage is when the cells level

out in numbers. -!%is staqe is the most important because this is when you have the most

cells in the tank. The last staqe is called the crashing stage. This is when the tank is over

run by a different diatom or when the tank decreases in cell densi-fy. Each staqe can

achieve optimal qrowth with different fertilizers. I was able to implement a fertilizinq -

schedule for achievinq - . optimal arowth -, (Figure #3).

** The MarBec Summer Research Pro.iect allowed me to have enouqh - time to iearn

about more than research, it allowed me to realize that this is a field that I would like t o

pursue in the future. Biotechnoloqy - . seems to be a fast qrowing field and my proJect gave

me a chance to learn more about it.

2 ' ' 5 7.- ."

I n particular, rnv project seemed to be like a puzzle. When you finally fiqure out

where one piece fits, there are still a iot o f other parts t o the puzzle. Even though

harvesting is a large part of culturing Chaetoceros there is stil l the raising, fertilizing and

the different stages the cells qo - through. Just like a puzzle, when I found where one

piece came together, I pushed on to find out where the rest of the pieces fit together.

PAGE 17

Acknowledqrnents

I would like to sav a special Thank You to several different people.

1) Sherwood Maynard and the Marine Opt~on Program: Thank you for all of the wonderful

help and support.

2) Sara Peck and the MarBec Summer Research Program: Thank you for allowing me to be able to accomplish all that T did this summer.

3) 01.. Jaw Kai Wanq: Thank you for the use of your facility at Anuenue Fisheries and for all the advise you were able to share with me,

4) Sally Koba: Thank you for all your heip with the equipment and for helping the proJect alonq with all the details that T missed.

5) Andrew Csorsa: Thank you for helpma - me piece toqether - all of my mixed thouqhts - and making sense out of them and for heipinq - me with ail of the problems that I had with mi: cro iect

8 6 ) J im Bish: Thank You for sharinq with me all of your expertise with the culturinq of the

dhaetoceros tanks and the use of all of your material for all of my crazy experiments.

6) Tom Ey' Thank You for the use of your facility at Anuenue Fisheries and for teaching me more of what aauaculture is al! about.

7 ) Vernon Sato: Thank You for giving me insight of the importance of my work and the experiments that I did.

8) Charley: Thank you for all of the technical advise and for the help with the building of the fractionator.

9) Joey: Thank You for helpinq . - me with all of the lab work.

Sheet4 Chart 1

Summer Chaetoceros Growth Chart - - - --

c u l t u r e #1 .Culture #2 Culture #3

lture

#2

Page 1

Cell Density

A A M N W W P m s n g g g g g

Weight in Grams

Weight in Grams