Evaluation of Hydrocyclone/UV Ballast Treatmentaboard the cruise ship SEA PRINCESS

• Moss Landing Marine Laboratories:Rusty FaireyNick WelschmeyerEli LandrauJudah GoldbergJason FeltonTam Voss

• Romberg-Tiburon Center, SanFrancisco State University

Steve BollensJena BillsSean AventTansy ClayRian Hoof

So…

Let’s test the system

Conclusions (at the moment):

• We find no significant effect from thehydrocyclone/UV treatment

• The largest measurable ‘effect’ derives fromthe ballast tank itself

Flow DiagramM/S Sea Princess

AZ

052

Micro

Kill S

ep M

od

. HR

N 200

UVControlPanel

MicroKill UV Mod:LP350-14- XF

AZ

056

BF

V 1

M1

CV

BFV 2

BFV 10

Pump C

AZ

054

Ovb. disc

M2

DN

50

mm

pur

ge li

ne

Main ballast line 200 mm CuNi 90/10

AZ

053

Pump B

AZ

055

AZ

051

AZ

034

AZ

????

AZ

????

Main bilge line 200 mm CuNi 90/10

BF

V 7

PowerSlave

BFV 8

BFV 9

DN50DN50

Red 150 x 200

BFV 5

Tee DN 200

Measurement parameters:

• Virus-like particles (VLP) – 0.02 um aluminum Anodisc filter, Syber-GoldStain, Epifluorescence enumeration

• Bacteria – same as VLP,• Phytoplankton – epifluorescence enumeration, bulk chl a (acetone extraction),

HPLC determined chlorophylls and carotenoids• Zooplankton – taxonomic abundance, survivorship• Pulse Amplitude Modulated Fluorescence (PAM) – Walz PAM 2000 (diode

detection); Walz Water PAM (photomultipier detection)• ATP – luciferin/luciferase, Turner 20/20 luminometer

• Others:Carbon/Nitrogen (POC, PON)Electron Transport System (ETS) respiratory activityBacteria - culture plates, nucleic acid ‘Live/Dead’ stainsGrow out experiments – phytoplankton, zooplankton

Diatom (red)

Bacteria(white)

DAPI stained Live/Dead stained

25 um

Live Bacteria(SYTO 9)

Dead Bacteria(Dead-Red)

Hydrocyclone/UV‘Microkill’ Treatment

ControlBallast Tank

TreatmentBallast Tank

Sea-to-Sea

Ocean

Ballast/De-Ballast

0.0E+00

2.0E+06

4.0E+06

6.0E+06

8.0E+06

1.0E+07

1.2E+07

1.4E+07

1.6E+07

1.8E+07

1

'Sea-to-Sea' Trials

Vir

use

s (V

LP

/mL

)

C T C T C T C T

0.0E+00

2.0E+05

4.0E+05

6.0E+05

8.0E+05

1.0E+06

1.2E+06

1.4E+06

1.6E+06

1.8E+06

1

'Sea-to-Sea' Trials

Bac

teri

a (c

ells

/mL

)

C T C TC T C T

Sea-to-Sea

0.000

0.050

0.100

0.150

0.200

0.250

0.300

0.350

0.400

0.450

1

Ch

loro

ph

yl a

(u

g/L

)

C T C TC T C T

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1

AT

P (

ug

/L)

C T C T C T C T

Sea-to-Sea

0.000

0.050

0.100

0.150

0.200

0.250

0.300

0.350

0.400

1

PA

M F

luo

resc

ence

(F

v/F

m)

Sea-to-Sea

C T

OptiMar Ballast System Efficacy (M/V Sea Princess Cruises 1 and 2): "Pre-Treatment" vs. "Post-Treatment"

0

10

20

30

40

50

60

70

80

90

Nauplii Calanoids Cyclopoids Harpacticoids Other Total

Taxa

% S

urv

ivo

rsh

ip (

Mea

n +

/- S

E)

Pre Treatment

Post Treatment

N = 10 experimentsn= # specimens countedn.s. = not significant

n.s . n.s . n.s . n.s . n.s . n.s .

0.0E+00

5.0E+06

1.0E+07

1.5E+07

2.0E+07

2.5E+07

3.0E+07

3.5E+07

1

Bac

teri

a (c

ells

/mL

)

B D B D B D B D

C

C

T

T

Ballast/De-Ballast Experiments

0.0E+00

2.0E+07

4.0E+07

6.0E+07

8.0E+07

1.0E+08

1.2E+08

1.4E+08

1

Vir

use

s (V

LP

/mL

)

B D B D B D B D

C

T

TC

Ballast/De-ballast

0.000

20.000

40.000

60.000

80.000

100.000

120.000

0.00

Ch

loro

ph

yll a

(u

g/L

)

B D B D B D B D

C

C

T

T

Ballast/De-ballast

0.000

0.050

0.100

0.150

0.200

0.250

0.300

0.350

0.400

0.450

0.500

1

PA

M F

luo

resc

ence

Yie

ld (

Fv/

Fm

)

C T

B D B D B D B D

C

TC

TBallast/De-Ballast

0.020

0.021

0.022

0.023

0.024

0.025

0.026

0.027

0.028

0.029

0.030

3 5 7 9 11 13 15 17 19 21 23

Elution Time (min)

VIS

Ab

sorb

ance

(re

lati

ve r

esp

on

se)

Chl c3

Peridinol

Chl c1,2

Peridinin

19' butanoyloxyfucoxanthin

Fucoxanthin

Diadinoxanthin

Zeaxanthin

beta-carotene

Chl a

0

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0 5 10 15 20 25 30

Elution Time (min)

Flu

ore

scen

ce (

rela

tive

)

Ballasting

De-Ballasting

Pheophorbide aPheophytin a

Conclusions (at the moment):

• We find no significant effect from thehydrocyclone/UV treatment

• The largest measurable ‘effect’ derives fromthe ballast tank itself

• We better do another test!!

Problems… More work…

Diatom (red)

Bacteria(white)

DAPI stained Live/Dead stained

25 um

Live Bacteria(SYTO 9)

Dead Bacteria(Dead-Red)