MERC ER02

- 10

ii 

# Land

- Based Evaluations of the

Siemens Water Technologies SiCURE TM



# Ballast Water Management System

Maritime(Environmental(Resource(Center



## September 2010

c/o Chesapeake Biological Laboratory

University of Maryland Center for Environm ental Science

PO Box 38, Solomons, Maryland 20688, USA

Ref. No. [UMCES]CBL 10

- 038MERC ER02

- 10

ii 

## Notic e

The objective of this Maritime Environmental Resource Center ( MERC ) evaluation wa s to provide shipping lines, classification societies, regulators

, and flag states with an independent

and credible assessment of treatment performance under realistic conditions. Therefore, the

ballast water treatment system was tested in accordance with the International Maritime

Organization (IMO) International Conven tion for the Control and Management of Ships’ Ballast

Water and Sediment (2004), Resolution MEPC.174(58) Guidelines for Approval of Ballast

Water Management Systems (G8) and Resolution MEPC.169(57) Procedure for Approval of

Ballast Water Management Systems T hat M ake U se of Active Substances (G9)

. The evaluation

was conducted under specific, predetermined, agreed

- upon protocols, criteria, and quality

assurance procedures to assess the treatment system's performance.

MERC does not label or list technologies a s acceptable or unacceptable but will present

the results in a n objective way that can be used to determine regulatory compliance by

appropriate administrations, agencies o r certification societies. Subsequent data on the

technology's performance characte ristics is presented to allow for compar ison with the IMO

Convention discharge standards, Regulation D

- 2

, Ballast Water Performance Standard

. MERC and the MERC Advisory Board do not provide certification for technologies, or

certify that a technology wil l always operate as demonstrated . Additionally, no expressed or

implied guarantee is provided as to the performance of the technology

, or that a technology will

always operate at the levels verified. MERC does guarantee the levels verified during the

eva luation under the conditions, circumstances , and operations encountered as fully independent

and credible.

This report has be en reviewed by members of the MERC Advisory Board and provided

to Siemens and MERC funding agencies prior to public release. Ment ion of trade names or

commercial products does not constitute endorsement or recommendation by MERC.

Questions and comments should be directed to Dr. Mario Tamburri, tamburri@umces.edu.MERC ER02

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iii 

### Table of Contents

Page No.

1.0. MERC Background and Objectives

................................ ................................ ............... 1

2

. 0. Description of the Siemens Ballast Water Management System

................................ ... 1

3.0. Summary of IMO Standards

................................ ................................ .......................... 2

4.0. Summary of Test Protocol s

................................ ................................ ............................ 2

5

. 0. Summary of Results

................................ ................................ ................................ ....... 4

6

. 0 Results

................................ ................................ ................................ ............................ 6

6

. 1. Trial 1 (SIE

- 03)

................................ ................................ ................................ .. 6

6

. 2. Trial 2 (SIE

- 05)

................................ ................................ ................................ .. 9

6.3 Trial 3 (SIE

- 06)

................................ ................................ ................................ 11

6.4. T rial 4 (SIE

- 07)

................................ ................................ ................................ 14

6.5. Trial 5 (SIE

- 08)

................................ ................................ ................................ 17

6.6. Twenty

- four Hour Dechlorination Trial

................................ .......................... 19

7

. Acknowledgements

................................ ................................ ................................ ...... 23

Appendix A. Vendor Interpretation

................................ ................................ ......................... 24MERC ER02

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### 1

. 0 MERC Background and Objectives

The Maritime Environmental Resource Center (MERC) is a State o f Maryland initiative

that provides test facilities, information, and decision tools to address key environmental issues

facing the international maritime industry. The Center’s primary focus is to evaluate the

mechanical and biological efficacy, associate d costs, and logistical aspects of ballast water

treatment systems and the economic impacts of ballast water regulations and management

approaches. A full description of MERC ’s structure, products, and services can be found at

www.maritime

- enviro.org.

To address the need for effective, safe, and reliable ballast water treatment systems to

prevent the introduction of non

- native species, MERC has developed as a partnership between

the Maryland Port Administration (MPA), Chesapeake Biological Laboratory/ Univ ersity of

Maryland Center for Environmental Science (CBL/UMCES), U.S. Maritime Administration

(MARAD), Smithsonian Environmental Research Center (SERC), and University of Maryland

(UM D ) to provide independent performance testing and to help facilitate t he transition of new

treatment technologies to shipboard implementation and operations.

This report describe s the MERC evaluation of the Siemens SiCURE TM 

### Ballast Water

Management System through objective and quality assured land

- based testing ( dockside at a

flow rate of 200m 3 

### /hr). The goal of this evaluation was to provide shipping lines, regulators,

classification societies, and flag states with an independent and credible assessment of treatment

performance under realistic conditions. Therefore, the data and information on performance

characteristics cover s legitimate information to meet the evaluation's objective, and performanc e is presented in a way to allow for comparison against the International Maritime Organization

(IMO ) International Convention f or the Control and Management of Ships' Bal last Water and

Sediments (2004 ) , Regulation D

- 2 Ballast Water Performance Standard

.

2. 0 Description of the Siemens Ballast Water Management System

The Siemens Water Technologies SiCURE TM 

### Ballast Water Managemen t System

(BWMS) utilizes a combination of treatment methods including physical separation and a

proprietary, on

- demand treatment with biocides produced in

- situ from seawater without the

addition of chemicals. This proprietary technology is based on the ma ritime industry

- proven

Chloropac® biofouling control technology that was first developed in the early 1970s and

installed on over 2,500 vessels.

The SiCURE TM 

### system employs electrolysis of seawater to produce a dilute solution of

sodium hypochlorite as an Active Substance that is injected into the ballast piping. The

proprietary controls regulate the system's parameters to provide only as much Active Substance

as required to achieve the necessary level of disinfection. This approach is aimed at eliminatin g

over

- chlorination

, associated risks of corrosion

, and generation of disinfection by

- products.

The specific SiCURE TM 

### unit tested as part of this MERC land

- based evaluation was a

prototype designed to land

- based evaluation conditions with shipboard impleme ntation

considerations. For this land based test, the water is first filtered through a 40 micron

BallastSafeTM BSFc Automatic Electric Filter, Model BSFc

- H

- 1.6 prior to treatment. The

sintered stainless steel screen technology enables it to remove zooplan kton. BallastSafe’s filter

features continuous cleaning of large volumes of dirt during ballasting without interruption, and

a reversible screw system for cleaning of the entire screen surface. This system requires theMERC ER02

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### discharge/backflus h of accumulated s olid residue (or retentate)

. To account for brackish water conditions

, the addition of a brine ( sodium chloride)

injection system was included for proper operation during periods of relatively low salinity. The

sodium hypochlorite solution was produce d fr om a small side stream of ballast water from the

main ballast piping. An advanced electrolyzer treated the side stream to create the required

Active Substance concentration on

- demand. The sodium hypochlorite / Active Substance solution

was then injected bo th before and after the filtration unit. The pre filltration injection accounted

for 10% or less of the total treatment dose for the purpose of filter biofouling prevention. An

injection quill specifically designed for this application was implemented fo r the primary dose

after the filter unit. The SiCURE TM 

### system doses at a maximum of 6 mg/l total chlorine. Upon

discharge of treated water, total chlorine levels are monitored and if the levels are greater than

the discharge limit of 0.1 ppm, an automate d system injects sodium sulfite as neutralizer.

Because the treatment system was a prototype, it was operated at all times by members of

the Siemens staff. All evaluation test equipment and instrumentation was operated by MERC

personnel.

3. 0 Summary of IMO Standards

This evalua tion was designed to determine i f the SiCURE TM 

### treatment system c ould meet

IMO D2 standards in accordance with both the IMO Guidelines for Approval of Ballast Water

Management Systems (G8) and the Procedure for Approval of Ballast Water Management

Systems that make use of Active Substances (G9) . The IMO Convention performance standard

states that ships must discharge:

1) Less than 10 viable organisms per m 3



### , greater than or equal to 50 μ m in minimum

dimension;

2) Less than 10 viabl e organisms per ml, less than 50 μ m in minimum dimension and greater

than or equal to 10 μ m in minimum dimension and

3) Less than the following concentrations of indicator microbes, as a human health standard:

1. Toxi genic Vibrio chol erae (serogroups O1 an d O139), less than 1 colony forming unit

(cfu) per 100 ml

2. Escherichia coli

, less than 250 cfu per 100 ml ;

3. I ntestinal Enterococci, less than 100 cfu per 100 ml

.

4

. 0 Summary of Test Protocols

The following is a brief summary of the testing approac h and methods. For complete

details on protocols, data management, and quality control / quality assurance procedures for this

MERC evaluation, please refer to the Test Plan for the Performance Evaluation of the S iemens

Ballast Water Management System ( Aug ust 2009 ), available for download at www.maritime

enviro.org

. The protocols described below are based upon the IMO G8/G9 guidelines and the U.S.

Coast Guard supported ETV protocols under development. Any deviation from IMO G8 /G9

guidelines or draft ETV p rotocols were explained and justified in the Test Plan.

MERC evaluated the biological efficacy of the SiCURE TM 

### ballast water treatment system

onboard the U.S. Maritime Administration ( MARAD ) Ro

- Ro vessel MV Cape Washington while

docked in the Port of Ba ltimore . The ballast system o n MV Cape Washington was modified toMERC ER02

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### allow for water at a flow rate of 4 00m 3 

### /hr to be split equally at flow rates of 200 m 3 

### /hr. Just

before this split, challenge condition concentrations of total suspended solids (TSS) and

par ticulate carbon (POC) were augmented by injecting a concentrated slurry of Arizona test dust

and humic acid (developed and validated by the Naval Research Laboratory, Key West, Florida).

The water was then delivered simultaneously to either a “control” (u ntreated) ballast tank or a

“treated” (passing first through the SiCURE TM 

### system) ballast tank. These two tanks were used

for the required holding time of five days and were essentially identical in size and structure.

Each tank was filled to approximately 250 m 3 

### for each test trial.

Physical Parameters

- Temperature, salinity, dissolved oxygen, chlorophyll fluorescence,

turbidity and pH were measured every 15 minutes during the test trials by two identical multi

parameter probes placed, one each, into the control and test tanks. Initial inline samples of ballast

water during the filling of the control and test tanks were collected, filtered, and analyzed (using

USEPA methods) for the water quality parameters of particulate organic carbon (POC),

dissolved o rganic carbon (DOC) a nd total suspended solids (TSS) by the CBL/UMCES Nutrient

A nalytical Services Laboratory (NASL).

Sampling

- A total of 10 identical 1.1 m 3 

### conical bottom mesocosms were also used for

controlled sampling during each trial. Using the me socosms, five sequential samples were taken

during: (A) initial filling of tanks, just prior to the split of control and treated water ( T0 Control ),

(B) initial filling of test tank, just downstream of the SiCURE TM 

### system during filling of test tank

( T0 Tr eated ), (C) during discharge of control water after a five

- day holding time ( TF Control ),

and (D) during discharge of treated water after a five

- day holding time ( TF Treated ).

Live Organisms > 50 μ m

- Immediately after filling, 1.0 m 3 

### of water in each me socosm

was filtered through a 35 μ m plankton net to concentrate the zooplankton for qualitative and

quantitative analyses under a dissecting microscope . T he proportion and total concentration of

live versus dead organisms was determined using standard mov ement and response

- to

- stimuli

techniques within one hour of collecting the individual samples. Zooplankton samples were also

fixed and returned to the laboratory for additional taxonomic evaluations.

Live Organisms 10

- 50 μ m

- Fifteen to twenty liters of well

- mixed, but unfiltered water

from each mesocosm were also collected immediately after filling, to determine concentrations

of organisms in the 10 to 50 micron size class using three distinct methods: (A) A sub

- sample

was stained using a combination of CMFDA (5

- chloromethylfluorescein diacetate) and FDA

(fluorescein diacetate) as selective live/viable indicators. Stained sub

- samples were incubated

and observed on a Sedgewick Rafter slide using a Leitz Laborlux S modified for epifluorescence

. (B) A s ec ond sub

- sample was fixed with standard Lugol's solution for use in taxonomic

identifications and to serve as a backup sample. (C) A third sub

- sample wa s filtered and frozen

until analysis of total and active chlorophyll

- a by the NASL

. (D) Finally , a fort h sub

- sample was

used to determine chlorophyll

- a levels after allowed to regrow under favorable conditions. An

increase in chlorophyll

, or positive regrowth

, indicates that viable phytoplankton were in the

samples, whereas chlorophyll levels at or below d etection limits of the laboratory analytical

method suggests that there was no viable phytoplankton.

Live Microbes

- Additional subsamples of unfiltered water were also collected from each

mesocosm to determine concentrations of total heterotrophic bacter ia and three specific indicator

pathogens, E. coli , intestinal Enterococci , and toxigenic Vibrio cholerae . Total heterotrophic

bacteria were enumerated by spread plate method using NWRI agar. The presence and

abundance of intestinal Enterococci was deter mined using a commercially availabl e chromogenic substrate method. C ulturable E. coli concentrations were determined using aMERC ER02

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### standard USEPA method: membrane filt ration on modified mTEC agar . Abundance s of total

and toxigenic V. cholerae were calculated by filtration and selection on TCBS agar and

enumerated using a sp ecies

- specific RNA colony blot and ctxA DNA colony blot hybridization

. Viable toxigenic V. cholerae was assayed with a commercial DFA kit specific for serogro up O1

using monoclonal antibodie s tagged w ith fluorescein isothiocyanate

. Toxicity

- To evaluate the toxicity of treated water at the completion of each trial, samples

from each mesocosm were collected and tested for chronic toxicity and for total residual

chlorine. Filter “backflush” ( retenate/filtrate during initial treatment of water) was also tested for

total residual chlorine. The toxicity protocols and species used were consistent with the US EPA

methods for Whole Effluent Toxicity (WET). The algal species tested was Isochrysis gal bana,

the fish species was the S heepshead minnow (Cyprinodon variegatus) while th e invertebrate

species was the M ysid shrimp (Americamysis bahia).

C hlorine concentration in samples was analyzed immediately upon collection to avoid

potential loss of oxida nt with time

. 

### The Standard Methods for the Examination of Water and

Wastewater Low

- Level Amperometric Titration method 4500

- Cl D and DPD Colorimetric

method 4500

- Cl G were used to measure Total Residual Oxidants (TRO ). 

### A Fischer and Porter

amperometric ti trator was also used for amperometric measurements.

To assess disinfection by

- products, a nine

- liter carboy of water was drawn at the

following 4 time

- points: T

- 0 Control inflow, T

- 0 Treated inflow, T

- 0 backflush discharge and T

F Treated discharge. The c arboy of water was immediately sampled for analyses of the

following compounds: Trihalomethanes (TTHM) , Haloa cetic Acids (HAA5) , Tribromoacetic

Acid (TBAA) , Bromate (Br) , Chlorate (ClO3) , Sodium ( Na, as a metal), and various Nitriles (

- CN group

– triple bo nd). Samples were iced and delivered same

- day via courier to the

Analytical Laboratory Services, Inc. (ALSI) for analysis.

5

. 0 Summary of Results

Live Organisms

- For essentially all biological categories, the Siemens SiCURE TM



### BWMS reduced the number s of live organisms in ballast water to levels below IMO D2

discharge standards. The only anomaly was found in one trial where Enterococci abundances

increased slightly in both treated and control water during the 5

- day hold time, with a T F value

just ove r ( 107.4 + 33.5 ) the IMO D2 discharge standard of less than 100 cfu/100 ml. However, it

is important to note the relatively large standard deviation, that the most probably number

( MPN ) method used (Idexx Enterolert kit) to quantify Enterococci commonly o ver estimate

abundance when compared to the a traditional membrane filtration method (G. Ditcher , personal

communication )

, and in all other test trials (over two different years) Enterococci appeared to be

successfully treated by the SiCURE TM 

### system.

Chr onic Toxicity

- Toxicity testing was conducted on five treated discharge samples

between 4/27/10 and 6/15/10. Results indicate that treated ballast water discharge (TF) was not

chronically toxic to mysid shrimp or Sheepshead Minnows. However the treated w ater upon

discharge did significantly inhibit the growth of the algae species

, Isochrysis galbana (T

- Iso. ) for

three of the test trials ( T rial 1 22

- 27 April

, Trial 2 13

- 18 May, and Trial 5 3

- 8 June ), which is

reported as “toxic” for this algal assay. Thes e samples had LOECs of 100% and NOECs of 56%.

All three algae tests with a toxicity response in the undiluted sample (100%) had similar EC50s

of 70%, 71% and 78% for SIE

- 03,

- 05 and

- 08, respectively.MERC ER02

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5 

### Residual Chlorine Analysis

- L evels of residual chlori ne (or Total R esidual O xidants

- TRO) were below the permissible discharge limit of 0.10 ppm in all TF treated samples. Values

ranged from 0.06 ppm to below the method detection limit (MDL) of 0.02 ppm. A summary

table from the completed trial runs and t he 24

- hour dechlorination run is provided below. TRO

was immediately measured shipboard using the DPD cholorimetric analysis method.

Note that Total Residual Chlorine (TRC) is not directly measured using the DPD

cholorimetric analysis method. The analy sis does not distinguish between various oxidants

(chlorine, bromine, ozone, etc). It reports TRO in chlorine equivalents (in other words, as if all

oxidant measured were chlorine). In freshwater, after introducing chlorine, we would be

comfortable calli ng the residual TRC. However, it is likely that in saltwater, while the majority

are chlorines, some portion may well be bromines. Thus, we only report TRO.

TRO as measured in the control and treated tanks.

Report ID Run ID Date TRO Treated

Tank TRO Con trol

Tank Comment

Trial 1 SIE

- 03

- 10 4/27 0.06 ppm

Trial 2 SIE

- 05

- 10 5/18 0.08 ppm 0.01 ppm

Trial 3 SIE

- 06

- 10 5/25 0.09 ppm 0.03 ppm

Trial 4 SIE

- 07

- 10 6/1 ND ND

Trial 5 SIE

- 08

- 10 6/8 0.06 ppm 0.04 ppm

24

- Hour

Trial SIE

- 09

- 10 6/10 0.59 ppm 0.06 ppm Initial

0.20 ppm Thiosulfate

0.21 ppm Thiosulfate

0.22 ppm Thiosulfate

0.04 ppm S ulfite

0.04 ppm S ulfite

0.02 ppm S ulfite

0.03 ppm S ulfite

0.06 ppm S ulfite

0.03 ppm S ulfite

Notes from dechlorination run.

1. Thiosulfate treatment did not result in adequate dechlorination at point of discharge.

2. Measured TRO levels in sodium sulfite treated samples were consistently below measure d free (active)

chlorine levels, suggestin g interference of sodium sulfite with DPD cholorimetric analysis.



### Disinfection By

- Products

- A summaries of the results for DBP analyses for each trial are

in the results section below. Reporting detection limits are provided.

Twenty four

- hour dechlorin tation trial

- On June 9

- 10, 2010, MERC conducted one

additional trial that required the use of the Siemens dechlorination system. While TF water in all

5

- day hold time trials had total chlorine levels below 0.1 ppm, a short 24

- hour hold time resulted

in measurable levels of chlorine and the subsequent automate dechlorination of the treated water

upon discharge using Sodium Sulfite (Na 2 

### SO 3 

### ,) . For this added trial, total chlorine levels in

discharged treated water declined in from 0.59 ppm prior to dechlor ination, to below 0.1 ppmMERC ER02

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### after the addition sodium bisulfate. Note an attempt at using sodium thiosulfate on discharge

was not effective, resulting in a final chlorine level of 0.21 ppm.

Mechanical Failures

- SIE

- 10

- 02

- T0 (15 April 2010) The filtration unit on the

SiCURE TM 

### system failed prior to the first sampling mesocosm being completely filled. The unit

clogged, causing a complete stoppage of flow. Trial SIE

- 10

- 02

- T0 was therefore canceled.

SIE

- 10

- 06

- T0 (20 May 2010) The SiCURE TM 

### system experienced a malfunction during startup

but prior to the filling of the treated tank and sampling mesocosms. Once the issue was resolved,

testing was re

- initiated. In all trials, 10

- 15 minutes of water flow through the system was

provided before electrochlorination was initiated . This flow was diverted into the a separated

ballast tanks and not into the treated test tank.

* Complete datasets and further performance information is available upon request.

6. 0 Results

6.1. Trial 1 (SIE

- 03

- 10): 22

- 27 April 2010

P resented as means and standard deviations.

Physical Parameters

TSS mg/l DOC mg/l PC mg/l

T0 Ambient 23.9 (3.7) 2.82 (0.01) 3.44 (0.03)

T0 Enhanced 51.3 (2.4) NA 5.61 (0.61)

Enhanced data is the average of 3 time points (2 reps per time point) duri ng the fill time of the ballast tanks.

Ambient water is sampled at the beginning of the trial and before the water is enhanced.

Temp o C Salinity psu Dis. Oxygen mg/l Turbidity NTU pH

T

- 0 Control 14.28 (0.03) 5.97 (0.00) 11.89 (0.03) 17.39 (1.10) 8.5 (0.1 )

T

- 0 Treated 14.36 (0.02) 6.64 (0.00) 16.24 (0.09) 18.46 (1.21) 8.6 (0.1 )

T

- F Control 14.14 (0.01) 5.95 (0.01) 8.37 (0.02) 3.67 (0.19) 7.8 (0.1 )

T

- F Treated 14.18 (0.01) 6.61 (0.01) 10.93 (0.01) 6.56 (0.09) 8.0 (0.1 )



### Live Organisms > 50 μ m

T0 #/m 3 T F #/m 3

Control 124,000 (5,000) *223,667 (29,501)

Treated 4.5 (7) 5 (4.5)

*Note large increase between T0 and TF in Control tank (reproduction and hatching of eggs).

*TF Control *TF Treated

Copepod nauplii Copepoda

Calanoida ( Acartia sp. ) Rot ifera

Cirripedia nauplii

Harpacticoida

* Up to four top t axa listed in order of abundance.MERC ER02

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### Live Organisms 10

- 50 μ m

Vital Stain Results

- It is important to note that the specific approach used underestimates

the abundances of live organisms in th is size class because it has been found that not all live

organisms reliably take up these stains.

T0 #/ml TF #/ml

Control 5

, 297 (776) 1

, 113 (225)

Treated 29 (19) 0 (0)

Dominant species Type Other

Heterocapsa rotundatum Dinoflagellate Small but a bundant

Gymnodinium sp. Dinoflagellate

Navicula sp. Diatom



### Active Chlorophyll

- a

- Chlorophyll is used as ancillary data and as a general

presence/absence indicator of viable photosynthetic organisms.

T0 μ g/l TF μ g/l RG μ g/l

Control 39.84 (11.7 7) 5.63 (0.50) 1.19 (0.13)

Treated 0.32 (0.13) 0.21 (0.02) 0.00 (0.01)

RG = after re

- growth assay ; MDL = 0.56 μ g/l



### Live Microbes

cfu = colony forming units



### E. coli T0 cfu/100ml TF cfu/100ml

Control 3.4 (1.52) 0.6 (1.34)

Treated 0 (0) 0 (0)



### Enteroc occi T0 NPM/ 100ml TF MPN/100ml

Control 3.92 (1.38) 9.06 (2.96)

Treated 1 (0.71) 1.2 (0.45)

MPN = most probably number estimate determined by the Idexx Enterolert kit.



### V. cholerae

– No detectable culturable toxigenic Vibrio cholerae were detected in an y

samples during any of the trials.

Heterotrophic

Bacteria T0 (sd)

cfu/1ml TF (sd)

cfu/1ml

Control 1,518 (373.6) 808 (190.66)

Treated 2.2 (3.58) 29,120 (2,184.49)MERC ER02

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8 

### Toxicity Summaries

Residual Chlorine Analysis

- Levels of residual chlorine (o r total residual oxidants

- TRO) were below the permissible discharge limit of 0.10 ppm in all TF treated samples. Values

ranged from 0.06 ppm to below the method detection limit (MDL) of 0.02 ppm.

Chronic Toxicity

- Results showed that ballast water dis charge was not chronically toxic

to either mysids or sheepshead minnows. All samples, however, were chronically toxic to the

marine algal species Isochrysis galbana with a statistically significant inhibition in growth in

treated ballast discharge compare d to controls.

Disinfection By

- Products ( μ g/l )

Control Treated BW Treated RDL/MRL

T0 T0 T0 TF

Trihalomethanes 0 91.2 ND 293 0.5

Trichloromethane < 0.5 < 0.5 ND < 0.5 0.5

Bromodichloromethane < 0.5 2.7 ND 3.8 0.5

Dibromochloromethane < 0.5 20.7 ND 28.2 0.5

Tribromomethane < 0.5 67.8 ND 261 0.5

Haloacetic Acids 0 87.4 ND 197 1.0

Monochloroacetic acid < 2.0 < 2.0 ND < 2.0 2.0

Dichloroacetic acid < 1.0 1.2 ND < 1.0 1.0

Trichloroacetic acid < 1.0 < 1.0 ND < 1.0 1.0

Bromochloroacetic acid < 1.0 5 ND < 1.0 1.0

Monobromoacetic acid < 1.0 4.5 ND < 1.0 1.0

Dibromoacetic acid < 1.0 72.2 ND < 1.0 1.0

Tribromoacetic acid < 1.0 4.5 ND 197 1.0

Other ND

Sodium chlorate < 10 340 ND 340 10

Sodium bromate < 50.0 < 50.0 ND < 50.0 50.0

Monochloroaceton itrile <0.5 <0.5 ND <0.5

Dichloroacetonitrile <0.5 <0.5 ND <0.5 0.5

Monobromoacetonitrile <0.5 <0.5 ND <0.5

Dibromoacetonitrile <0.5 18 ND <0.5 0.5

BW = backwash; RDL = reporting detection limit ; MRL = method reporting l i mitMERC ER02

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9 

### 6.2. Trial 2 (SIE

- 05

- 1 0): 13

- 18 May 2010

Presented as means and standard deviations.

Physical Parameters

TSS mg/l DOC mg/l POC mg/l

T0 Ambient 11.5 (1.0) 3.41 (0.13) 1.11 (0.03)

T0 Enhanced 65.2 (8.5) NA 5.88 (0.79)

Enhanced data is the average of 3 time points (2 reps per time point) during the fill time of the ballast tanks.

Ambient water is sampled at the beginning of the trial and before the water is enhanced.

Temp o C Salinity psu Dis. Oxygen mg/l Turbidity NTU pH

T

- 0 Control 15.15 (0.01) 8.19 (0.02) 8.54 (0. 06) 15.60 (0.87) 7.3 (0.1 )

T

- 0 Treated 15.20 (0.01) 8.84 (0.01) 9.02 (0.02) 15.11 (1.24) 7.3 (0.1)

T

- F Control 16.53 (0.06) 8.15 (0.00) 6.92 (0.06) 3.49 (0.23) 7. 2 (0. 2 )

T

- F Treated 16.52 (0.05) 8.76 (0.00) 8.74 (0.03) 5.27 (0.42) 7.1 (0.1)



### Live Organ isms > 50 μ m

T0 #/m 3 * T F #/m 3

Control 60,000 (10,500) 90,500 (10,000) ŧ

Treated 4.2 (7) 0.2 (0.4)

*Note moderate increase between T0 and TF in Control tank (reproduction and hatching of eggs).

*TF Control *TF Treated

Copepod nauplii Polycheata (Spionidae)

Calanoida (Acartia sp.)

Polychaeta (Spionidae)

Harpacticoida

* Up to four top t axa listed in order of abundance.



### Live Organisms 10

- 50 μ m

Vital Stain Results

- It is important to note that the specific approach used underestimates

the abundances of live organisms in this size class because it has been found t hat not all live

organisms reliably take up these stains.

T0 #/ml TF #/ml

Control 1,915 (511) 3,182 (725)

Treatment 1.7 (1.2) 6 (3.8)

Dominant species Type Other

Heterocapsa rotundatum Dinoflagellate

Gymnodinium sp. Dinoflagellate

Thalassiosir a gravida Diatom chain

- forming

Navicula sp. DiatomMERC ER02

- 10

10 

### Active Chlorophyll

- a

- Chlorophyll is used as ancillary data and as a general

presence/absence indicator of viable photosynthetic organisms.

T0 μ g/l TF μ g/l RG μ g/l

Control 8.79 (0.48) 1.89 (0.06) 5.95 (3.11)

Treated 0.08 (0.01) 0.03 (0.01) 0.08 (0.09)

RG = after re

- growth assay

MDL = 0.56 μ g/l



### Live Microbes

cfu = colony forming units



### E. coli T0 cfu/100ml TF cfu/100ml

Control 4.6 (1.82) 0 (0)

Treated 0 (0) 1 (1.41)



### Enterococci T0 MPN/ 100ml TF MPN/100ml

Control 8.78 (21.2) 49.34 (26.54)

Treated 2.02 (1.45) 107.4 (33.5)

MPN = most probable number estimate s determined by the Idexx Enterolert kit. Note that this MPN is not a direct

cfu count and Enterolert MPN values are com monly found to be higher than the traditional membrane filtration

method used to estimate Enterococci abundances (G. Ditcher, Idexx).



### V. cholerae

– No detectable culturable toxigenic Vibrio cholerae were detected in any

samples during any of the trials.

Heterotrophic

Bac teria T0 cfu/1ml TF cfu/1ml

Control 146 (80) 503.33 (187.75)

Treated 0 (0) 10,520 (4,447.13)



### Toxicity Summaries

Residual Chlorine Analysis

- Levels of residual chlorine (or total residual oxidants

- TRO) were below the permissible discharge limit of 0.10 ppm in all TF treated samples. Values

ranged from 0.06 ppm to below the method detection limit (MDL) of 0.02 ppm.

Chronic Toxicity

- Results showed that ballast water discharge was not chronically toxic

to either mysids or sheepshead minnows. Al l samples, however, were chronically toxic to the

marine algal species Isochrysis galbana with a statistically significant inhibition in growth in

treated ballast discharge compared to controls.MERC ER02

- 10

11 

### Disinfection By

- Products ( μ g/l)

Control Treated BW Treated RDL/MRL

T0 T0 T0 TF

Trihalomethanes 0 106.6 ND 227.8 0.5

Trichloromethane < 0.5 < 0.5 ND < 0.5 0.5

Bromodichloromethane < 0.5 1.6 ND 2.5 0.5

Dibromochloromethane < 0.5 13.5 ND 24.3 0.5

Tribromomethane < 0.5 91.5 ND 201 0.5

Haloacetic Acids 1.4 6 2.9 ND 385.6 1.0

Monochloroacetic acid < 2.0 < 2.0 ND < 2.0 2.0

Dichloroacetic acid < 1.0 < 1.0 ND < 1.0 1.0

Trichloroacetic acid < 1.0 < 1.0 ND < 1.0 1.0

Bromochloroacetic acid < 1.0 3.3 ND < 1.0 1.0

Monobromoacetic acid < 1.0 3.1 ND 1 1.0

Dibromoac etic acid < 1.0 53 ND 10.6 1.0

Tribromoacetic acid 1.4 3.5 ND 374 1.0

Other ND

Sodium chlorate < 10 250 ND 250 10

Sodium bromate < 50.0 < 50.0 ND < 50.0 50.0

Monochloroacetonitrile <0.5 <0.5 ND <0.5

Dichloroacetonitrile <0.5 0.86 ND <0.5 0.5

Mo nobromoacetonitrile <0.5 <0.5 ND <0.5

Dibromoacetonitrile <0.5 24 ND 4.4 0.5

BW = backwash

RDL = reporting detection limit

MRL = method reporting l i mit



### 6

. 3. Trial 3 (SIE

- 06

- 10): 20

- 25 May 2010

Presented as means and standard deviations.

Physical Pa rameters

TSS mg/l DOC mg/l POC mg/l

T0 Ambient 19.7 (3.6) 2.96 (0.00) 1.28 (0.01)

T0 Enhanced 56.2 (13.5) NA 4.76 (0.92)

Enhanced data is the average of 3 time points (2 reps per time point) during the fill time of the ballast tanks.

Ambient water is sampled at the beginning of the trial and before the water is enhanced.

Temp o C Salinity psu Dis. Oxygen mg/l Turbidity NTU pH

T

- 0 Control 15.14 (0.02) 9.80 (0.02) 6.47 (0.39) 15.68 (1.30) 7.1 (0.1 )

T

- 0 Treated 15.18 (0.00) 10.35 (0.00) 7.91 (0. 01) 14.95 (1.01) 7.0 (0. 1 )

T

- F Control 19.83 (0.01) 9.75 (0.00) 4.82 (0.02) 3.06 (0.10) 7.1 (0. 1 )

T

- F Treated 19.97 (0.01) 10.37 (0.00) 7.55 (0.01) 6.61 (0.09) 7.0 (0. 1 )MERC ER02

- 10

12 

### Live Organisms > 50 μ m

T0 #/m 3 T F #/m 3

Control 50,500 (10,000) 35,666 (2,35 0)

Treated 9 (7) 9 (3.5)

*TF Control *TF Treated

Copepod nauplii Bivalve larvae

Polychaeta (Spionidae) Copepoda

Calanoida ( Acartia sp, Eurytemora sp .) Turbellaria

Harpacticoida

* Up to four top t axa listed in order of abundance.



### Live Organisms 10

- 50 μ m

Vital Stain Results

- It is important to note that the specific approach used underestimates

the abundances of live organisms in this size class because it has been found that not all live

organisms reliably take up these stains.

T0 #/ml TF #/ml

Control 1,088 (194) 137 (8)

Treatment 1.7 (1.2) 0 (0)

Dominant species Type Other

Heterocapsa rotundatum Dinoflagellate

Gymnodinium sp. Dinoflagellate

Thalassiosira gravida Diatom chain

- forming

Navicula sp. Diatom



### Active Chlorophyll

- a

- C hlorophyll is used as ancillary data and as a general

presence/absence indicator of viable photosynthetic organisms.

T0 μ g/l TF μ g/l RG μ g/l

Control 3.07 (0.09) 0.91 (0.02) 0.12 (0.09)

Treated 0.24 (0.17) 0.13 (0.02) 0.00 (0.00)

RG = after re

- growt h assay ; MDL = 0.56 μ g/l



### Live Microbes

cfu = colony forming units



### E. coli T0 cfu/100ml TF cfu/100ml

Control 0 (0) 2.2 (1.5)

Treated 0 (0) 0 (0)



### Enterococci T0 MPN/ 100ml TF MPN/100ml

Control 16.64 (4.08) 70.42 (29.77)

Treated 5.8 (1.9) 34.68 (8.5 9)

MPN = most probably number estimate determined by the Idexx Enterolert kit.MERC ER02

- 10

13 

### V. cholerae

– No detectable culturable toxigenic Vibrio cholerae were detected in any

samples during any of the trials.

Heterotrophic

Bacteria T0 cfu/1ml TF cfu/1ml

Control 950 (300) 55 (31)

Treated 0 (0) 13,780 (2,560)



### Toxicity Summaries

Residual Chlorine Analysis

- Levels of residual chlorine (or total residual oxidants

- TRO) were below the permissible discharge limit of 0.10 ppm in all TF treated samples. Values

ra nged from 0.06 ppm to below the method detection limit (MDL) of 0.02 ppm.

Chronic Toxicity

- Results showed that ballast water discharge was not chronically toxic

to any of the three tested species.

Disinfection By

- Products ( μ g/l)

Control Treated BW Tr eated RDL/MRL

T0 T0 T0 TF

Trihalomethanes 0 27 0.63 168.2 0.5

Trichloromethane < 0.5 < 0.5 < 0.5 < 0.5 0.5

Bromodichloromethane < 0.5 1.2 < 0.5 2.3 0.5

Dibromochloromethane < 0.5 10.6 < 0.5 17.9 0.5

Tribromomethane < 0.5 15.2 0.63 148 0.5

Haloace tic Acids 0 71.1 1.9 6.9 1.0

Monochloroacetic acid < 2.0 < 2.0 < 2.0 < 2.0 2.0

Dichloroacetic acid < 1.0 < 1.0 < 1.0 < 1.0 1.0

Trichloroacetic acid < 1.0 < 1.0 < 1.0 1.4 1.0

Bromochloroacetic acid < 1.0 2.9 < 1.0 < 1.0 1.0

Monobromoacetic acid < 1.0 1 .4 < 1.0 < 1.0 1.0

Dibromoacetic acid < 1.0 28.4 < 1.0 5.5 1.0

Tribromoacetic acid < 4.0 38.4 1.9 0 1.0

Other

Sodium chlorate < 10 170 < 10 < 10 10

Sodium bromate < 50.0 < 50.0 < 50.0 < 50.0 50.0

Monochloroacetonitrile <0.5 <0.5 <0.5 <0.5

Dich loroacetonitrile <0.5 <0.5 <0.5 <0.5 0.5

Monobromoacetonitrile <0.5 <0.5 <0.5 <0.5

Dibromoacetonitrile <0.5 15 <0.5 7.4 0.5

BW = backwash

RDL = reporting detection limit

MRL = method reporting l i mitMERC ER02

- 10

14 

### 6

. 4. Trial 4 (SIE

- 07

- 10): 27 May

– 1 June 2010

Pr esented as means and standard deviations.

Physical Parameters

TSS mg/l DOC mg/l POC mg/l

T0 Ambient 12.7 (0.4) 3.16 (0.02) 1.29 (0.04)

T0 Enhanced 57.9 (1.7) NA 5.36 (0.19)

Enhanced data is the average of 3 time points (2 reps per time point) duri ng the fill time of the ballast tanks.

Ambient water is sampled at the beginning of the trial and before the water is enhanced.

Temp o C Salinity psu Dis. Oxygen mg/l Turbidity NTU pH

T

- 0 Control 17.40 (0.03) 8.81 (0.03) 7.09 (0.10) 15.84 (1.11) 6.9 (0.1)

T

- 0 Treated 17.48 (0.02) 9.42 (0.00) 5.47 (0.04) 16.09 (1.21) 7 .0 (0.1)

T

- F Control 19.53 (0.01) 8.87 (0.00) 5.18 (0.10) 2.22 (0.08) 7.2 (0.1)

T

- F Treated 19.69 (0.01) 9.48 (0.00) 5.57 (0.02) 6.07 (0.13) 7.2 (0.1)



### Live Organisms > 50 μ m

T0 #/ m 3 T F #/m 3

Control 113,333 (14,154) 63,500 (9,750)

Treated 13 (12) 1.8 (0.8)

*TF Control *TF Treated

Calanoida ( Eurytemora sp. ) Copepoda

Copepod nauplii Bivalve larva

Polychaeta (Spionidae)

Bivalve larvae

* Up to four top t axa listed in order o f abundance.



### Live Organisms 10

- 50 μ m

Vital Stain Results

- It is important to note that the specific approach used underestimates

the abundances of live organisms in this size class because it has been found that not all live

organisms reliably take u p these stains.

T0 #/ml TF #/ml

Control 1,544 (103) 1,026 (133)

Treatment 1 (1) 1.2 (0.8)

Dominant species Type Other

Prorocentrum minimum Dinoflagellate

Heterocapsa rotundatum Dinoflagellate

Gymnodinium estuarale Dinoflagellate first time det ected in 2010

Thalassiosira sp. Diatom chain

- forming

Navicula sp. DiatomMERC ER02

- 10

15 

### Active Chlorophyll

- a

- Chlorophyll is used as ancillary data and as a general

presence/absence indicator of viable photosynthetic organisms.

T0 μ g/l TF μ g/l RG μ g/l

Control 6.42 (0.78) 1.08 (0.04) 1.41 (0.47)

Treated 0.05 (0.01) 0.02 (0.01) 0.00 (0.00)

RG = after re

- growth assay

MDL = 0.56 μ g/l



### Live Microbes

cfu = colony forming units



### E. coli T0 cfu/100ml TF cfu/100ml

Control 2.2 (2.49) 0 (0)

Treated 0 (0) 0 (0)



### En terococci T0 MPN/ 100ml TF MPN/100ml

Control 10 (4) 11.62 (3.75)

Treated 2.46 (2.16) 57.96 (30.54)

MPN = most probably number estimate determined by the Idexx Enterolert kit.



### V. cholerae

– No detectable culturable toxigenic Vibrio cholerae were detecte d in any

samples during any of the trials.

Heterotrophic

Bacteria T0 cfu/1ml TF cfu/1ml

Control 470 (176) 190 (63.07)

Treated 1.4 (1.43) 7,630 (4,181)



### Toxicity Summaries

Residual Chlorine Analysis

- Levels of residual chlorine (or total residual oxidants

- TRO) were below the permissible discharge limit of 0.10 ppm in all TF treated samples. Values

ranged from 0.06 ppm to below the method detection limit (MDL) of 0.02 ppm.

Chronic Toxicity

- Results showed that ballast water discharge was not ch ronically toxic

to any of the three tested species.MERC ER02

- 10

16 

### Disinfection By

- Products ( μ g/l)

Control Treated BW Treated RDL/MRL

T0 T0 T0 TF

Trihalomethanes 0 98.2 0 281.2 0.5

Trichloromethane < 0.5 < 0.5 < 0.5 < 0.5 0.5

Bromodichloromethane < 0.5 1.6 < 0 .5 2.6 0.5

Dibromochloromethane < 0.5 14.9 < 0.5 25.6 0.5

Tribromomethane < 0.5 81.7 < 0.5 253 0.5

Haloacetic Acids 0 56.7 0 117.6 1.0

Monochloroacetic acid < 2.0 < 2.0 < 2.0 < 2.0 2.0

Dichloroacetic acid < 1.0 < 1.0 < 1.0 < 1.0 1.0

Trichloroacetic a cid < 1.0 < 1.0 < 1.0 < 1.0 1.0

Bromochloroacetic acid < 1.0 4.1 < 1.0 < 1.0 1.0

Monobromoacetic acid < 1.0 3.4 < 1.0 < 1.0 1.0

Dibromoacetic acid < 1.0 49.2 < 1.0 10.6 1.0

Tribromoacetic acid < 4.0 0 < 4.0 107 1.0

Other

Sodium chlorate < 10 180 < 10 240/200 10

Sodium bromate < 50.0 < 50.0 < 50.0 < 50.0 50.0

Monochloroacetonitrile <0.5 <0.5 <0.5 <0.5

Dichloroacetonitrile <0.5 <0.5 <0.5 <0.5 0.5

Monobromoacetonitrile <0.5 <0.5 <0.5 <0.5

Dibromoacetonitrile <0.5 26 <0.5 12 0.5

BW = backwash

RDL = reporting detection limit

MRL = method reporting l i mitMERC ER02

- 10

17 

### 6

. 5. Trial 5 (SIE

- 08

- 10): 3

- 8 June 2010

Presented as means and standard deviations.

Physical Parameters

TSS mg/l DOC mg/l POC mg/l

T0 Ambient 17.1 (5.1) 3.76 (0.11) 1.56 (0.18)

T0 Enhanced 55.0 (5.6) NA 5.39 (0.52)

Enhanced data is the average of 3 time points (2 reps per time point) during the fill time of the ballast tanks.

Ambient water is sampled at the beginning of the trial and before the water is enhanced.

Temp o C Sal inity psu Dis. Oxygen mg/l Turbidity NTU pH

T

- 0 Control 20.48 (0.02) 7.41 (0.00) 4.82 (0.15) 15.96 (1.13) 7.1 (0.1)

T

- 0 Treated 20.52 (0.01) 6.99 (0.01) 7.89 (0.01) 16.05 (1.14) 7.1 (0.1)

T

- F Control 20.13 (0.02) 7.50 (0.00) 3.13 (0.04) 1.32 (0.07) 7.0 (0.1)

T

- F Treated 20.27 (0.03) 6.34 (0.01) 7.94 (0.00) 7.22 (0.18) 7.0 (0.1)



### Live Organisms > 50 μ m

T0 #/m 3 T F #/m 3

Control 175,000 (11,000) 94,500 (8,250)

Treated 221 (59) 8 (3)

*TF Control *TF Treated

Copepod nauplii Copepoda

Bivalve larvae Polychaeta (Spionidae)

Spionidae

Calanoida ( Eurytemora sp .)

* Up to four top t axa listed in order of abundance.



### Live Organisms 10

- 50 μ m

Vital Stain Results

- It is important to note that the specific approach used underestimates

the abundances o f live organisms in this size class because it has been found that not all live

organisms reliably take up these stains.

T0 #/ml TF #/ml

Control 1,511 (271) 757 (145)

Treatment 1.7 (1.2) 2.2 (1.5)

Dominant species Type Other

Prorocentrum minimum D inoflagellate

Thalassiosira sp. Diatom c hain

- forming

Gyrodinium estuarale Dinoflagellate

Navicula sp. Diatom

Amphora sp. DiatomMERC ER02

- 10

18 

### Active Chlorophyll

- a

- Chlorophyll is used as ancillary data and as a general

presence/absence indicator of viable pho tosynthetic organisms.

T0 μ g/l TF μ g/l RG μ g/l

Control 3.42 (0.12) 0.44 (0.14) 0.97 (0.36)

Treated 0.04 (0.01) 0.01 (0.00) 0.00 (0.00)

RG = after re

- growth assay

MDL = 0.56 μ g/l



### Live Microbes

cfu = colony forming units



### E. coli T0 cfu/100ml TF cfu /100ml

Control 13.4 (5.5) 0 (0)

Treated 0 (0) 0 (0)



### Enterococci T0 MPN/ 100ml TF MPN/100ml

Control 113.94 (38.9) 2.42 (0.58)

Treated 1.02 (1.45) 1 (1)

MPN = most probably number estimate determined by the Idexx Enterolert kit.



### V. cholerae

– No det ectable culturable toxigenic Vibrio cholerae were detected in any

samples during any of the trials.

Heterotrophic

Bacteria T0 cfu/1ml TF cfu/1ml

Control 577 (149) 466 (671)

Treated 2.3 (2.5) 3,950 (884)



### Toxicity Summaries

Residual Chlorine Analys is

- Levels of residual chlorine (or total residual oxidants

- TRO) were below the permissible discharge limit of 0.10 ppm in all TF treated samples. Values

ranged from 0.06 ppm to below the method detection limit (MDL) of 0.02 ppm.

Chronic Toxicity

- Re sults showed that ballast water discharge was not chronically toxic

to either mysids or sheepshead minnows.

All samples, however, were chronically toxic to the marine algal species Isochrysis

galbana with a statistically significant inhibition in growth in treated ballast discharge compared

to controls.MERC ER02

- 10

19 

### Disinfection By

- Products ( μ g/l)

Control Treated BW Treated RDL/MRL

T0 T0 T0 TF

Trihalomethanes 0 241.4 0 89.6 0.5

Trichloromethane < 0.5 < 0.5 < 0.5 < 0.5 0.5

Bromodichloromethane < 0.5 2.6 < 0.5 3.3 0.5

Dibromochloromethane < 0.5 119.8 < 0.5 24.5 0.5

Tribromomethane < 0.5 119 < 0.5 61.8 0.5

Haloacetic Acids 0 143.8 0 95.6 1.0

Monochloroacetic acid < 2.0 < 2.0 < 2.0 < 2.0 2.0

Dichloroacetic acid < 1.0 < 1.0 < 1.0 < 1.0 1.0

Trichloroacetic aci d < 1.0 < 1.0 < 1.0 < 1.0 1.0

Bromochloroacetic acid < 1.0 3.5 < 1.0 < 1.0 1.0

Monobromoacetic acid < 1.0 3.2 < 1.0 < 1.0 1.0

Dibromoacetic acid < 1.0 37.1 < 1.0 8.9 1.0

Tribromoacetic acid < 4.0 100 < 4.0 86.7 1.0

Other

Sodium chlorate < 10 295 /240 < 10 < 10/297 10

Sodium bromate < 50.0 < 50.0 < 50.0 < 50.0 50.0

Monochloroacetonitrile <0.5 <0.5 <0.5 <0.5

Dichloroacetonitrile <0.5 <0.5 <0.5 <0.5 0.5

Monobromoacetonitrile <0.5 <0.5 <0.5 <0.5

Dibromoacetonitrile <0.5 24 <0.5 8.5 0.5

BW = ba ckwash

RDL = reporting detection limit

MRL = method reporting l i mit



### 6

. 6. Twenty four

- hour Dechlorination Trial: 9

- 10 June 2010

Presented as means and standard deviations.

Physical Parameters

TSS mg/l DOC mg/l POC mg/l

T0 Ambient 33.4 ( 9.6 ) 3.11 (0.12) 2.06 (0.09)

T0 Enhanced NA NA NA

Enhanced data is the average of 3 time points (2 reps per time point) during the fill time of the ballast tanks.

Ambient water is sampled at the beginning of the trial and before the water is enhanced.

Temp o C Salinity psu Dis. Oxygen mg/l Turbidity NTU pH

T

- 0 Control 21.03 (0.08) 8.39 (0.02) 6.38 (0.22) 3.22 (0.12) 7.5 (0.1)

T

- 0 Treated 21.15 (0.04) 9.01 (0.01) 7.35 (0.02) 7.53 (0.20) 7.0 (0.1)

T

- F Control 20.51 (0.04) 8.41 (0.00) 5.76 (0.03) 2.96 (0.14) 7 .0 (0.1)

T

- F Treated 20.74 (0.03) 9.04 (0.00) 7.32 (0.01) 7.29 (0.10) 7.0 (0.1)MERC ER02

- 10

20 

### Live Organisms > 50 μ m

T0 Av g(sd) #/m 3 T F Av g(sd) #/m 3

Control 160,500 (9,750) 112,500 (5,000)

Treated 17 (7) 1.2 (2)

* Up to four top t axa listed in order of abunda nce.



### Live Organisms 10

- 50 μ m

Vital Stain Results

- It is important to note that the specific approach used undere stimates

the abundances of live organisms in this size class because it has been found that not all live

organisms reliably take up these stains.

T0 #/ml TF #/ml

Control 4,593 (823) 3,355 (149)

Treatment 0 (0) 0 (0)

Dominant species Type Other

Pro rocentrum minimum Dinoflagellate

Thalassiosira sp. Diatom chain

- forming

Gyrodinium estuarale Dinoflagellate

Cyclotella glomerata Diatom chain

- forming

Skeletonema costatum Diatom chain

- forming

Heterocapsa triquerta Dinoflagellate

Navicula sp. Diat om

Nitzschia sp. Diatom



### Active Chlorophyll

- a

- Chlorophyll is used as ancillary data and as a general

presence/absence indicator of viable photosynthetic organisms.

T0 μ g/l TF μ g/l RG μ g/l

Control 12.33 (0.77) 9.66 (0.16) NA

Treated 0.04 (0.01) 0.01 (0.00) NA

RG = after re

- growth assay

MDL = 0.56 μ g/l * TF Control * TF Treated

Copepod nauplii Copepoda

Bivalve larvae Polychaeta (Spionidae)

Spionidae

Calanoida ( Eurytemora sp .)MERC ER02

- 10

21 

### Live Microbes

cfu = colony forming units



### E. coli T0 cfu/100ml TF cfu/100ml

Control 10.33 (9.29) 2.67 (2.52)

Treated 0 (0) 0 (0)



### Enterococci T0 MPN/ 100ml TF MPN/100ml

Control 5.13 (2.05) 8.9 (2.88)

Treated 0.33 (0.58) 0.33 (0.58)

MPN = most probably number estimate determined by the Idexx Enterolert kit.



### V. cholerae

– No detectable culturable toxigenic Vibrio cholerae were detected in any

samples during any of the trials.

Heterotrophi c

Bacteria T0 cfu/1ml TF cfu/1ml

Control 2,180 (383) 631.67 (218.68)

Treated 3.5 (6.06) 0 (0)



### Toxicity Summaries

Residual Chlorine Analysis

– On June 9

- 10, 2010, MERC assisted Siemens with the

testing of their dechlorination system. A 24

- hour tri al resulted in successful dechlorination of

the treated tank using sodium bisulfate (a decline in total chlorine levels from 0.59 ppm to below

0.1 ppm

– the EPA discharge standard). Note that using a sodium thiosulfate treatment resulted

in a final total chlorine level of 0.22 ppm.

Chronic Toxicity

– B allast water was tested after treatment with a higher concentration of

chlorine followed by an in

- line de

- chlorination. Ballast water was collect in the same manner

and location as previous tests. All thre e species (mysid, fish and algae) were tested for toxicity.

There was no statistically significant toxicological effect seen in any of these three test species.MERC ER02

- 10

22 

### Disinfection By

- Products ( μ g/l)

Control Treated BW Treated RDL/M

RL

T0 T0 T0 TF

Trihal omethanes 0 88.4 0 238.9 0.5

Trichloromethane < 0.5 < 0.5 < 0.5 < 0.5 0.5

Bromodichloromethane < 0.5 1.4 < 0.5 2.1 0.5

Dibromochloromethane < 0.5 15 < 0.5 22.8 0.5

Tribromomethane < 0.5 72 < 0.5 214 0.5

Haloacetic Acids 0 118.7 0 193.3 1.0

Monochloro acetic acid < 2.0 < 2.0 < 2.0 < 2.0 2.0

Dichloroacetic acid < 1.0 < 1.0 < 1.0 < 1.0 1.0

Trichloroacetic acid < 1.0 < 1.0 < 1.0 < 1.0 1.0

Bromochloroacetic acid < 1.0 2.5 < 1.0 3.1 1.0

Monobromoacetic acid < 1.0 2.5 < 1.0 3.8 1.0

Dibromoacetic acid < 1 .0 25 < 1.0 39.4 1.0

Tribromoacetic acid < 4.0 88.7 < 4.0 147 1.0

Other

Sodium chlorate < 10 276/340 < 10/390 233/330 10

Sodium bromate < 50.0 < 50.0 < 50.0 < 50.0 50.0

Monochloroacetonitrile <0.5 <0.5 <0.5 <0.5

Dichloroacetonitrile <0.5 <0.5 < 0.5 <0.5 0.5

Monobromoacetonitrile <0.5 <0.5 <0.5 <0.5

Dibromoacetonitrile <0.5 28 <0.5 29 0.5

BW = backwash

RDL = reporting detection limit

MRL = method reporting l i mitMERC ER02

- 10

23 

### 7

. 0 Acknowledgments

The MERC Testing Team included J. Barn es, M. Carroll, D. Fisher, B. Haley, A. Huq, R.

Kanzleiter, T. Mullady, G. Ruiz, G. Smith, D. Spar ks, M. Tamburri. E. Taviani, M. Wilkinson,

L. Yonkos, and G. Ziegler. Test protocols were developed in conjunction with A. Cangelosi, the

Great Ships Initiative, Bundesamt fuer Seeschifffahrt und Hydrographie (BSH) and M. Veldhuis

. We wish thank the crew of the MV Cape Washington who provided critical input and support o n all aspects of these trials, and the Maryland Port Administration and U.S. Maritime

Administration for fund ing and supporting this ballast water treatment evaluation.

16 September 2010

16 September 2010

Date Approved By: Dr. Mario Tamburri

MERC Executive Director

Date Approved By: Ross Kanzleiter

MERC Program Coordinator and

Chief Engineer