Ballast Water Sampling and Analysis Workshop - Balticcomplete

Ballast Water Sampling and Analysis Workshop

COMPLETE Workshop

08-10 May 2019, Hamburg, Germany

Katja Broeg, German Federal Maritime and Hydrographic Agency, BSH

CONTENT

Summary

Invitation

Agenda

Workshop Material

Questionnaire: Challenges for the participating countries

List of Participants

1

SUMMARY Participants from administrations and scientific sectors of seven Baltic Sea countries who are going

to perform ballast water sampling and analysis met in a COMPLETE workshop organised in

Hamburg at the beginning of May. For the first time, knowledge and practical hands-on exercises

for all stages from planning and sampling to analysis were included in one seminar, which was

supervised by the internationally recognised scientists Dr. Matej David and Dr. Stephan Gollasch.

Starting with background knowledge on sampling and analysis tools and methods, the participants

got the opportunity to carry out practical work in a laboratory and on board the TUI cruise liner

„Mein Schiff 5“. All sections of the workshop took place right at the Elbe River, opposite to the

Port of Hamburg that celebrated its 830th birthday in parallel. The gathered expertise will be

further distributed by the participants, who also brought in their experience and questions

concerning the further development of all ballast water issues. The materials of the workshop are

provided by the COMPLETE project. In addition, a documentary on the practical exercises

combined with interviews with the experts will be prepared.

Figure 1: Dr. Stephan Gollasch demonstrates the refractometer for measuring the salinity. This device is used for monitoring the D-1 standard (ballast water exchange).

2

Figure 2: Big interest for the investigation of the pankton sample. For many prticipants the first visual contact with the small water organisms which are in the centre of the BWMC.

Figure 3: Viable of dead? This is the question which must be answered for the evaluation of the D-2 standard. All which is glowing here due to staining with a flourescent dye, is living.

3

Figure 4: For others a place to relax, for many of the participants of the workshop the place of their first active ballast water sampling practice.

Figure 5: Participants from seven Baltic Sea countries will spread their gathered knowledge further #BalticComplete

08/05/2019 1 www.balticcomplete.com

Workshop on sampling and analysis of ballast water

Dr. Matej David and Dr. Stephan Gollasch

Programme

Wednesday, 8th

May 2019 morning: arrival of participants

Theme: BW Sampling

13:30-14:00 Arrival of participants and set-up in the meeting room

Introduction of participants

14:00-14:45 Ballast water sampling under the BWM Convention

14:45-15:30

Ballast water sampling (BWS) recommendations for compliance monitoring and

enforcement (CME) under the BWM Convention (ballast water exchange standard (D-1),

ballast water performance standard (D-2), indicative and detailed sampling, in-tank and at

discharge sampling, introduction of sampling gear, representativeness, sample volumes,

timing, number of samples

15:30-16:00 Coffee Break

16:00-17:00 Ballast water sampling recommendations for compliance control under the BWM

Convention - continues

Thursday, 9th

May 2019 Day 2

Theme: BW Sampling - Summary

09:00-09:15 Questions and answers from previous day

09:15-10:30 Selecting vessels and tanks for BWS for CME

10:30-11:00 Coffee break

Theme: BW Sample Analysis - Theory

11:00-11:15 Ballast water sample handling and transfer to the laboratory

11:15-11:30 Sample analysis procedures or techniques used to test for compliance with D-1

11:30-12:00 Practical handling of D-1 compliance tools

12:00-14:00 Lunch Break

14:00-15:30 Sample analysis procedures or techniques used to test for compliance with D-2, indicative

and detailed sample processing, introduction of sample analysis methods

15:30-16:00 Coffee break

16:00-16:45 Discussion

16:45-17:00 Planning of next day.

08/05/2019 2 www.balticcomplete.com

Friday, 10th

May 2019 Day 3

BW Sampling - Practical Part The test vessel is in walking distance to the meeting venue

09:00-09:15 Questions and answers from previous day

09:15 Split in two groups

Group 1 (ca. 10 participants)

Group 2 (ca. 10 participants)

09:30-12:00

Leave meeting venue for vessel visit Sampling for D-1 and D-2 compliance on a

ship

in tank sampling,

in-line sampling,

using D-1 analysis tools on board Dr. Matej David

Practical sample analysis for D-2 in the

laboratory Familiarization with microscopes Dr. Stephan Gollasch

12:00-13:30 Lunch break

13:30-16:00

Practical sample analysis for D-2 in the

laboratory Familiarization with microscopes Dr. Stephan Gollasch

Leave meeting venue for vessel visit Sampling for D-1 and D-2 compliance on a

ship

in tank sampling,

in-line sampling,

using D-1 analysis tools on board Dr. Matej David

16:00-17:00 Debrief of the vessel visit and the sampling on board Discussion Closing of meeting

Departure of participants

© 2002 By Default!

A Free sample background from www.powerpointbackgrounds.com

Slide 1

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Slide 2

➢BWM Convention articles relevant for Ballast Water Sampling (BWS)

➢BWS requirements under BWM Convention

➢Consequence of non-compliance

➢BWS under G2 Guidelines

➢BWS Guidance

➢BWS under PSC Gudelines

➢Conclusions

OUTLINE

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Slide 3

BWM Convention➢Article 9 Inspection of Ships

➢Article 10 Detection of Violations and Control of Ships

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Slide 4

➢Tier 1:

➢ valid certificate

➢ check BW record book

➢ BWS, conducted according to G2

BWM Convention & BWS Compliance control (Article 9)

➢ no undue delay because of

samples analysis

➢ no prevention of discharge

before having results of BWS

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Slide 5

➢ no valid certificate; or

➢ clear grounds…

➢ Paragraph 2

(Article 9.2)

➢ BWMS not correspond to certificate

➢ crew not familiar with BWM

procedures or not implemented

➢ Consequences?

➢ Tier 2: detailed inspection

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Slide 6

Consequence?

Article 9.3

➢ In the circumstances given in paragraph 2 of this Article, the Party carrying out the inspection shall take such steps as will ensure that the ship shall not discharge Ballast Water until it can do so without presenting a threat of harm to the environment, human health, property or resources.

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Slide 7

➢ indicative test shown “high”

organism load – clear indication or

grounds

➢ detailed sampling inspection

(in a previous port) – failed compliance

BWS was conducted

➢ Consequences?

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Slide 8

Article 10.3

➢ If the sampling described in Article 9.1(c) leads to a result, or supports information received from another port or offshore terminal, indicating that the ship poses a threat to the environment, human health, property or resources, the Party in whose waters the ship is operating shall prohibit such ship from discharging Ballast Water until the threat is removed.

Consequence?

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Slide 9

…but what to do with such vessel?

➢ no discharge

➢ port reception facility

➢ designated discharge/contingency area

➢ if none of these?

…how such vessel can prove again to be

compliant?

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Slide 10

BWS UNDER THE

G2 GUIDELINES

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Slide 11

Indicative test (G2, 6.3)

➢ Prior to testing for compliance with the D-2

standard, it is recommended that, as a first step, an

indicative analysis of ballast water discharge may be

undertaken to establish whether a ship is potentially

compliant or non-compliant.

➢ Such a test could help the Party identify immediate

mitigation measures, within their existing powers, to

avoid any additional impact from a possible non-

compliant ballast water discharge from the ship.

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Slide 12

➢ the sampling protocol should result in samples that are representative of the whole discharge of ballast water from any single tank or any combination of tanks being discharged;

➢ the sampling protocol should take account of the potential for a suspended sediment load in the discharge to affect sample results;

➢ the sampling protocol should provide for samples to be taken at appropriate discharge points;

➢ the quantity and quality of samples taken should be sufficient to demonstrate whether the ballast water being discharged meets with the relevant standard;

BWS for compliance (G2, 6.2) -

Detailed test

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Slide 13

G2 continues…➢ sampling should be undertaken in a safe and

practical manner;

➢ samples should be concentrated to a manageable size;

➢ samples should be taken, sealed and stored to ensure that they can be used to test for compliance with the Convention;

➢ samples should be fully analysed within test method holding time limit using an accredited laboratory; and

➢ samples should be transported, handled and stored with the consideration of the chain of custody.

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Slide 14

WHERE CAN WE SAMPLE?

➢ In-tank ➢ In-line (at discharge)

➢ Which sampling point is appropriate?

➢ Sampling pointsavailability?

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Slide 15

BALLAST WATER SAMPLING

THAT IS

REPRESENTATIVE

OF THE

WHOLE DISCHARGE

➢ probably the biggest challenge…

➢many questions and issuess… how to do it?

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Slide 16

➢How to sample for D-1?

➢How to sample for D-2?

➢What is a representative sample?

➢What should be the sample volume?

➢How many samples to take?

➢Where to take samples?

➢When to take samples?

➢Which sampling gear to use?

➢ …

Clear guidance needed how

to do BWS for CME!

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Slide 17

BWS GUIDANCE

© 2002 By Default!


Slide 18

BWM.2/Circ.42 Guidance on ballast watersampling and analysis for trial use inaccordance with the BWM Convention andGuidelines (G2) (2013, rev.1 2015)

➢ list of sample analysis protocols,methodologies and approaches for D-1 and D-2 standards compliance tests

➢ 2 - 3 year trial period for reviewing, improvingand standardizing the BWM Circular

➢ recommendations for a trial period duringwhich sampling experience is gained

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Slide 19

CME UNDER THE

PSC GUIDELINES

(“G15”)

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Slide 20

➢ PSC Guidelinesadopted at MEPC67(Oct 2014)

➢ Provide basicguidance for PSCinspection to verifycompliance with therequirements of theBWM Convention

Guidelines for Port State Control

under the BWM Convention

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Slide 21

1 - "initial inspection" documentation check (IBWMC,

BWMP, BWRB), visual check of equipment,

ensuring that an officer has been nominated for

BWM on the ship, crew familiar to operate BWMS

if clear grounds… then…

2 - "more detailed inspection" where the operation of

the BWMS is checked and the PSCO clarifies

whether the BWMS has been operated adequately,

may result in sampling

3 - "indicative analysis", sampling for compliance with

D-2, no undue delay because of analysis

4 - "detailed analysis", sampling for compliance with

D-2, no undue delay while waiting for sample

analysis results

Four-stage inspection

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Slide 22

Four-stage inspection

order?➢ BWS under the BWM Convention as Tier 1

➢ BWS under G15 as third step as part of/or triggeredby a (more) detailed inspection

➢ conflict ?... BW discharge not allowed when Tier 2 =detailed inspection (Art. 9.2)! Art9.3 …ship shall notdischarge when Art. 9.2 conditions = detailed BWScan’t be conducted.

“Compromise”?:

1.1.1 These Guidelines… …are not intended to limit the

rights the port State has in verifying compliance with

the BWM Convention.

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Slide 23

As an alternative to warning, detention or exclusion of the

ship:

➢ retention of all ballast water on board;

➢ require the ship to undertake any repairs required tothe BWMS;

➢ permit the ship to proceed to exchange ballast water ina location acceptable to the port State;

➢ allow the ship to discharge ballast to another ship or toan appropriate shipboard or land-based receptionfacility; or

➢ allow the ship to manage the ballast water or a portionof it in accordance with a method acceptable to the portState.

➢ notify the ship and the flag State of the violation

Control actions

when non-compliant

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Slide 24

➢ BWM Convention introduces BWS as one ofthe methods for CME

➢ G2 Guidelines do not describe BWS in detail

➢ Correct BWS for CME is very important alsobecause of possible consequences for vesselsand environment

➢ BWS Guidance introduces a transitional periodimportant also for PSC to test and introducereliable BWS process for CME

➢ PSC Guidance introduces four step process,but there may be some implementationissues/conflicts

CONCLUSIONS

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Slide 25

MORE INFO IN THE NEW BOOK:

David, M., Gollasch, S. (eds.) 2015. Global

Maritime Transport and Ballast Water

Management – Issues and Solutions.

Springer Science and Business media

For more in information please visit:

http://www.springer.com/environment/environme

ntal+management/book/978-94-017-9366-7

The second edition of the book is in preparation with different new

subjects and lessons learnt important for the implementation of the

BWM Convention. Publication is expected in 2019.

http://www.springer.com/environment/environmental+management/book/978-94-017-9366-7

© 2002 By Default!


Slide 26

Dr. Stephan Gollasch

www.gollaschconsulting.de/

Prof. Dr. Matej David

www.davidconsult.eu/

http://www.gollaschconsulting.de/

http://www.davidconsult.eu/

THANK YOU

FOR THE

ATTENTION!

QUESTIONS

?

© 2002 By Default!


Slide 28

© 2002 By Default!


Slide 1

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Slide 2

➢D-1 sampling background and recommendations

➢D-2 sampling background and recommendations

➢Sampling gear and sampling point arrangements

➢Personal protective equipment

➢Conclusions

OUTLINE

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Slide 3

Recommendations based on:➢Test voyage 2009

➢Test voyages 2010

➢Test voyage 2012

➢ BALMAS 2013-2016

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Slide 4

➢ More than 1500 samples taken during more then 150 shipboard tests for type approval of >20different BWMS which were conducted by S. Gollasch and M. David over the past 12 years

Experience summarised in the study for WWF (March 2013):➢ Ballast water sampling for

compliance monitoring -Ratification of the Ballast Water Management Convention (MEPC 65/2/17)

© 2002 By Default!


Slide 5

SAMPLING FOR D-1

© 2002 By Default!


Slide 6

SAMPLING PURPOSECompliance with D-1 standard - Ballast Water Exchange

Standard

CHECK IF REQUIRED MANAGEMENT WAS CONDUCTED!

Regulation D-1

➢ BWE efficiency of at least 95 % volumetric exchange

Regulation B-4.1

➢ BWE >200 NM from the nearest land & water depth of at

least 200 m; …IF NOT POSSIBLE THEN

➢ BWE > 50 NM from the nearest land & water depth 200 m;

© 2002 By Default!


Slide 7

Samping Guidance

© 2002 By Default!


Slide 8

What to do in terms of

sampling?➢ Salinity check

➢ Small quantity needed

Where to sample?➢ In-tank

➢ In-line (at discharge) – sampling point needed…

© 2002 By Default!


Slide 9

SALINITY CHECK

Basic principle:

➢ Salinity at “open sea” ~ and >35 psu

➢ If vessel conducts BWE according tho the BWM Convention (95% exchange >50NM & 200m), ballast water should always have salinity >30 psu

➢ Method mainly limited to vessels that ballasted in low salinity areas.

© 2002 By Default!


Slide 10

16,1

17,2

19,2

15

16

17

18

19

20

10 14 16

Salin

ity [

psu

]

Depth [m]

In-tank sampling

Ballast water sampled from 3 different depths - different salinity measured

➢ Salinity stratification identified!

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Slide 11

In-line sampling

Ballast water sampled in 5 sequences during the discharge -different salinity measured

➢ Salinity stratification identified!

31,6

28,5

17,1

6,4 6,0

0

5

10

15

20

25

30

35

10 20 50 65 70

Sa

lin

ity [

ps

u]

Time [min.]

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Slide 12

IN-TANK

Recommended sampling approach for D-1

Sampling point Equipment Water

volume

Number of samples

Sounding pipe,

manhole or air

vent

Column sampler

or pump

> 50 ml 1 integrated sample

from possibly whole

water column

Sounding pipe,

manhole or air

vent

Column spot

sampler or

pump


from 3 different

depths

© 2002 By Default!


Slide 13

IN-LINE to be conducted if in-tank sampling not possible!

Sampling point Equipment Water volume Number of

samples

In-line Sampling jar or

bottle

> 50 ml 1 sample as soon

as possible* during

the discharge

* note:

➢ important to take it as soon as possible to prevent possible

discharge non-compliant water

➢ more samples may be taken with ~10 min delay to identify if

salinity is droping throught the time of discharge

© 2002 By Default!


Slide 14

D-2 SAMPLING

© 2002 By Default!


Slide 15

SAMPLING PURPOSECompliance with D-2 standard - Ballast Water Performance

Standard

➢ <10 organisms > 50 µm/m³

➢ <10 organisms <50 & > 10µm/ml

➢ Indicator microbes➢ Escherichia coli <250 cfu in 100 ml

➢ Enterococci <100 cfu in 100 ml

➢ Vibrio cholerae <1 in 100 ml

CHECK FOR THE CONCENTRATION OF VIABLE ORGANISMS AND BACTERIA (CFU) BEING DISCHARGED!

© 2002 By Default!


Slide 16

INDICATIVE SAMPLING

D-2

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Slide 17

WHERE TO SAMPLE?

➢ Sample in-tank (avoid discharge of BW)

➢ No in-tank sampling points installed on vessel

➢ Sampling possible also in-line (discharge started, indication still faster then detailed test)

SAMPLING PURPOSE➢ Recommended prior to detailed sampling to avoid any

additional impact from a possible non-compliant BW➢ In case tanks have direct discharge to the environment

© 2002 By Default!


Slide 18

TESTS OF BWS & A

➢ BALMAS testing of different sampling approaches for the D1 and indicative D-2 CME test using BBE 10 cells

➢ Sampling over time for salinity and organisms concentrations from three different depths + microscop counts

© 2002 By Default!


Slide 19

© 2002 By Default!


Slide 20

BBE

mean

Salinity Tempera

ture

day 1 tank 1 BW top 42.7

BW middle 84.3

BW bottom 91.7

intank mixed 86.3

tank 2 BW top 226.3

BW middle 255.3

BW bottom 552.3

intank mixed 345.3

day 2 tank 1 BW top 24.7 32.1 15.7

BW middle 51.3 32.2 14.1

BW bottom 80.7 32.6 13.9

intank mixed 56.0

tank 2 BW top 173.0 32.4 13.8

BW middle 250.0 32.4 13.0

BW bottom 558.0 33.6 12.9

intank mixed 342.3

day 3 tank 1 BW top 35.7 32.3 14.7

BW middle 54.7 32.3 14.1

BW bottom 71.7 32.2 13.6

intank mixed 56.0

tank 2 BW top 237.7 32.4 14.3

BW middle 301.0 32.4 14.3

BW bottom 570.7 32.3 14.0

intank mixed 385.3

day 4 tank 1 BW top 44.0 31.9 8.5

BW middle 57.0 32.0 7.4

BW bottom 63.0 32.0 6.7

intank mixed 46.3

tank 2 BW top 444.3 32.2 8.7

BW middle 498.0 32.2 7.5

BW bottom 682.7 32.2 6.7

intank mixed 521.7

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Slide 21

INDICATIVE SAMPLINGAPPROACHES

Principle 1: Prevent possible non-compliant discharge

➢ Only one group of organisms is enough to indicate/identify non-compliance!

➢ Phytoplankton indicative analysis tools (PAM based) are practical for onboard use and need low water quantity

➢ Zooplankton sample may be concentrated onboard and brought to a laboratory for fast analysis

➢ no appropriate indicative analysis tool for bacteria?

Principle 2: Indicative test may be followed/expanded in the detailed test

➢ Possibly sample as it would be the first part of the detailed test (include all groups of organisms)

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Slide 22

PRINCIPLE 1Organism

group

Sampling point Equipment Water volume

[litre]

Number of samples

< 50 and > 10

micrometres

Sounding pipe,

manhole or air

vent

Column

sampler or

pump


from possibly the

whole water column or

from 3 different depths

Sounding pipe,

manhole or air

vent

Column spot

sampler or

pump



IN

SUMMARY

-

IN-TANK

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Slide 23

IN

SUMMARY

-

IN-TANK

Organism

group

Sampling point Equipment Water volume

[litre]

Number of samples

> 50

micrometres

Manhole Plankton net 300 - 500 1 integrated sample

from possibly the

whole water column

Manhole, sounding pipe

or air vent

Pump 100 1 integrated sample

from possibly the



< 50 and > 10

micrometres

Manhole, sounding pipe,

manhole or air vent

Pump or water

column sampler

5 – 6

(1 enough)

1 integrated sample

from possibly the




manhole or air vent

Pump or point-

source sampler

5 – 6

(1 enough)

1 integrated sample


Indicator

microbes


manhole or air vent

Pump or water

column sampler

1 1 integrated sample

from possibly the

whole water column


manhole or air vent

Pump or point-

source sampler

1 1 integrated sample


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Slide 24

PRINCIPLE 2

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Slide 25

IN

SUMMARY

-

IN-LINE

Organism

group

Sampling

point

Equipment Water

volume

[litre]

Number of samples

> 50

micrometres

In-line Plankton net 300 – 500 1 sequential sample of ca.

10 minute duration,

avoiding the very

beginning and very end of

the tank discharge event

< 50 and >

10

micrometres

In-line Sampling jar and

bucket

5 – 6 1 continuous drip sample

may be simultaneously

collected during sampling

of organism group > 50

micrometres

Indicator

microbes

In-line Bucket, sampling

jar and bottle

1 1 continuous drip sample

may be sub-sampled from

the bucket

© 2002 By Default!


Slide 26

DETAILED SAMPLING-

COMPLIANCE CONTROL SAMPLING

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Slide 27

SAMPLE REPRESENTATIVENESS IS KEY

➢ Where to sample?

➢ When should sampling start and end?

➢ How long to sample?

➢ How many samples to take?

➢ How much volume to sample?

SAMPLING PURPOSE➢ Identify / accertain if the vessel is in compliance with the

requirements of the BWM Convention D-2 standard – i.e., viable organisms concentrations in the discharged water

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Slide 28

Compliance Control Sampling

➢ Voyages on commercial vessels

➢ Water flow split equally (untreated water)

– split 1 sample taken over entire pumping time

(OET), i.e. the whole discharge

– split 2 sequential samples in ca.

beginning, middle and end of the

pumping

event

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Slide 29

TEST VOYAGE 2009 (BSH)5 min. 10 min. 15 min. 5 min. 75 min.

TEST VOYAGES 2010 (EMSA)

Blue = sampling time

White = no sampling

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Slide 30

TEST VOYAGE 2012 (BSH)

➢Continuous sequences (no time gap

between sequences)

➢OET

➢ 4 tests

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Slide 31

➢ Organisms patchiness occurs

➢ No trend identified

➢ Similar results OET and sequences


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Slide 32



➢ Much higher concentrations in the sequencial samples


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Slide 33



➢ Similar concentration of organisms in sequances and OET


70 70

51

169

26

79

144

246

122

95

112

56

23

114

186

0

50

100

150

200

250

300

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

1 2 4 5

Sequences number organisms

10 - 50 µm / ml

Over entire timenumber

organisms10 - 50 µm / ml

© 2002 By Default!


Slide 34


➢ Rise of organism concentration from beginning to the end

➢ Higher organisms concentration in sequences vs. OET


18671956

2459

982

1368

1679

1190

1524

970

1259

2111

2667

1153

268

10681218

0

500

1000

1500

2000

2500

3000

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

DISCH, S1

DISCH, S2

DISCH, S3

DISCH, OET

1 2 4 5


50 µm and above / m3

Over entire time number

organisms 50 µm and above / m3

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Slide 35

➢ Higher concentrations in the OET samples

➢ Highest concentration varied


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Slide 36

➢ Higher concentrations in the sequencial samples vs. OET

➢ Highest concentration in end sequence


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Slide 37

STUDY COMPARISONOrganisms Study 2009 Study 2010 Study 2012

< 50 and > 10 µm

- Distribution

- Organism numbers

- Comparison

sequences/OET

Patchy

No trend per

sequence number

Similar numbers

Patchy

No trend per

sequence number

Similar numbers

Patchy

No trend per

sequence number

In OET higher

numbers

> 50 µm

- Distribution

- Organism numbers

- Comparison

sequences/OET

Patchy

No trend per

sequence number

In sequences much

higher numbers

Patchy

Increase towards

last sequence

In sequences much

higher numbers

Patchy

Highest

concentration in

last sequence

In sequences much

higher numbers

© 2002 By Default!


Slide 38

WHERE TO SAMPLE?

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Slide 39

➢ Sample in-line (at discharge) as this representsdischarge of organisms to the environment

➢ In-tank sampling represents potential to be discharged (organisms may remain in tank with remaining water) therefore it is difficult to prove discharge standard (need to reveal very high org. numbers – e.g. 1000 org. sampled from a tank of 100 m3)

➢ Sampling point needs to be installed on vessel

➢ Sampling bin and discharge provided on vessel (more about under “sampling arrangements”)

WHERE TO SAMPLE?

© 2002 By Default!


Slide 40

WHEN SHOULD SAMPLING

START AND END?

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Slide 41

Homogeneity of organism concentrations

in the dischargeOrganisms < 50 and > 10 – sequences compared

70 70

51

20 16

8879

144

246

122

95

112

0

50

100

150

200

250

300

S1 S2 S3 S1 S2 S3 S1 S2 S3 S1 S2 S3

1 2 4 5

Discharge sequences number

organisms 10 - 50 µm / ml

➢Organisms patchiness occurs

➢ No pattern identified

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Slide 42

Organisms > 50 – sequences compared


in the discharge


➢ Rise of organism concentration from beginning to the end

18671956

2459

982

1368

1679

1190

1524

970

1259

2111

2667

0

500

1000

1500

2000

2500

3000

S1 S2 S3 S1 S2 S3 S1 S2 S3 S1 S2 S3

1 2 4 5



© 2002 By Default!


Slide 43

Organisms > 50 & < 50 and > 10 – sequences compared

➢ Clear tank organisms patchiness

➢ BW sampling results clearly impacted by tank patchiness!


in the discharge

18671956

2459

982

1368

1679

1190

1524

970

1259

2111

2667

700 700

510

200 160

880790

1444

2457

1222

951

1123

0

500

1000

1500

2000

2500

3000

S1 S2 S3 S1 S2 S3 S1 S2 S3 S1 S2 S3

1 2 4 5



Discharge sequences 10 x number


© 2002 By Default!


Slide 44

Sampling timing

➢Do not start sampling during first 5 minutes after start of BW discharge

➢Do not sample in the last 5 minutes before end of discharge

Reasoning:

➢High patchiness of organisms during that sampling times; and

➢More sediment present in that sampling periods what may negatively affect organisms survival in the sample and sample processing.

© 2002 By Default!


Slide 45

HOW LONG TO SAMPLE?

© 2002 By Default!


Slide 46

BW sampling Over Entire Time vs.

Sequential ➢Organisms < 50 and > 10 – sequences compared

➢Similar results OET and Sequences

75

60 6370 70

51

9 717 16

9

2620 16

88

193

212217

79

144

246

122

157163

122

95

112

77

56

3123

47

230

114

223

186

0

50

100

150

200

250

300

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

1 1 2 2 3 4 4 5 5


10 - 50 µm / ml

Over entire timenumber

organisms10 - 50 µm / ml

© 2002 By Default!


Slide 47

Organisms > 50 – sequences compared

➢Much higher concentrations in the sequence samples

➢ Longer time & higher volume sampling negatively impacts

5096

45044237

18671956

2459

16891911

1067

386737

982 9821368

1679

24292762

2143

11901524

970

8370

4593

7444

1259

2111

2667

2004

1153373 268 749

1701

1068

2688

1218

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

UP

T, S

1

UP

T, S

2

UP

T, S

3

UP

T, O

ET

DIS

CH

, S1

DIS

CH

, S2

DIS

CH

, S3

DIS

CH

, OET

1 1 2 2 3 4 4 5 5


50 µm and above / m3

Over entire time number


BW sampling Over Entire Time vs.

Sequential

© 2002 By Default!


Slide 48

Representativeness (…of the whole discharge of ballast water from any single tank or any combination of tanks = ?

…representative sample in diversity, concentration and viability of organisms)

“STATISTICAL”

representativeness

“BIOLOGICAL”

representativeness

?

© 2002 By Default!


Slide 49

Sampled time

STATISTICAL

representativeness

BIOLOGICAL

representativeness

lower higher

➢missing diversity

➢ higher patchiness

representative

lower higher

➢ low confidence

empty e.g. 10.000 m3

➢ high confidence

➢ more stress

➢ organisms die

representative

© 2002 By Default!


Slide 50

➢ “BIOLOGICAL” representativeness is crucial!

➢ Statistical representativeness also to be considered

© 2002 By Default!


Slide 51

Sampling time duration

➢Recommended sampling time is ~10 minutes

Reasoning:

➢ Longer sampling time negatively affects survival of organisms > 50, hence sample is underestimating “real” organisms concentration; and

➢Shorter sampling times are still representativein organisms group < 50 and > 10.

© 2002 By Default!


Slide 52

HOW MANY SAMPLES TO TAKE?

© 2002 By Default!


Slide 53

Organisms > 50 & < 50 and > 10 – sequences compared

➢ Clear tank organisms patchiness

➢ BW sampling results clearly impacted by tank patchiness!


in the discharge

18671956

2459

982

1368

1679

1190

1524

970

1259

2111

2667

700 700

510

200 160

880790

1444

2457

1222

951

1123

0

500

1000

1500

2000

2500

3000

S1 S2 S3 S1 S2 S3 S1 S2 S3 S1 S2 S3

1 2 4 5



Discharge sequences 10 x number


© 2002 By Default!


Slide 54

“Instantaneous” sampling vs. “Average”Definition of instantaneous sampling:

➢ “one-point-in-time” sampling

➢Shorter sampling time than OET

Definition of average sampling:

➢Average of two or more instantaneous samples, or

➢OET sampling

© 2002 By Default!


Slide 55

Organisms < 50 and > 10 – 3 sequences vs. OET compared

“Instantaneous” sampling vs. “Average”

Test No. Sample type Total

number

organisms

10 - 50

µm / ml

Difference

between

sequence

and OET

(%)

DISCH, 3 SEQ AV 64 14

DISCH, OET 56

DISCH, 3 SEQ AV 41 -12

DISCH, OET 47


DISCH, OET 114


DISCH, OET 186

4

5

1

2

Result:

➢ Similar probability to sample higher or lower concentration

➢ Similar difference in organism concentrations sampled

© 2002 By Default!


Slide 56


“Instantaneous” sampling vs.

“Average”


number

organisms

10 - 50

µm / ml

Difference

between

sequence

and OET

(%)

DISCH, S1 70 25

DISCH, S2 70 25

DISCH, S3 51 -9


DISCH, OET 56

DISCH, S1 20 -57

DISCH, S2 16 -66

DISCH, S3 88 87


DISCH, OET 47

DISCH, S1 79 -30

DISCH, S2 144 27

DISCH, S3 246 116


DISCH, OET 114

DISCH, S1 122 -34

DISCH, S2 95 -49

DISCH, S3 112 -40


DISCH, OET 186

2

4

5

1

Method – Middle concentration sequence excluded

Result:

➢ Similar probability to sample higher or lower concentration

➢ Similar difference in concentrations sampled

© 2002 By Default!


Slide 57



“Average”


number

organisms

10 - 50

µm / ml

Difference

between

sequence

and OET

(%)

DISCH, S1 70 25

DISCH, S2 70 25

DISCH, S3 51 -9


DISCH, OET 56

DISCH, S1 20 -57

DISCH, S2 16 -66

DISCH, S3 88 87


DISCH, OET 47

DISCH, S1 79 -30

DISCH, S2 144 27

DISCH, S3 246 116


DISCH, OET 114

DISCH, S1 122 -34

DISCH, S2 95 -49

DISCH, S3 112 -40


DISCH, OET 186

1

2

4

5

Method – beginning or end

sequence excluded

Result:

➢ Similar probability to

sample higher or lower

concentration

➢ Similar difference in

concentrations sampled

© 2002 By Default!


Slide 58

Organisms > 50 – 3 sequences vs. OET compared

Result:

➢Always much higher organism concentration in sequences average


“Average”

Test No. Sample type Discharge

sequences

number

organisms

50 µm and

above / m3

Difference

between

sequence

and OET

(%)


DISCH, OET 1153


DISCH, OET 749


DISCH, OET 1068


DISCH, OET 1218

4

5

1

2

© 2002 By Default!


Slide 59



“Average”

Method - Second highest

sequence excluded

Result:

➢Always much higher

organism concentration

in sequences average


number

organisms

10 - 50

µm / ml

Difference

between

sequence

and OET

(%)

DISCH, S1 1867 62

DISCH, S2 1956 70

DISCH, S3 2459 113


DISCH, OET 1153

DISCH, S1 982 31

DISCH, S2 1368 83

DISCH, S3 1679 124


DISCH, OET 749

DISCH, S1 1190 11

DISCH, S2 1524 43

DISCH, S3 970 -9


DISCH, OET 1068

DISCH, S1 1259 3

DISCH, S2 2111 73

DISCH, S3 2667 119


DISCH, OET 12185

1

2

4

© 2002 By Default!


Slide 60


Method – beginning or end

sequence excluded

Result:

➢Always much higher

organism concentration

in sequences average


“Average”


number

organisms

10 - 50

µm / ml

Difference

between

sequence

and OET

(%)

DISCH, S1 1867 62

DISCH, S2 1956 70

DISCH, S3 2459 113


DISCH, OET 1153

DISCH, S1 982 31

DISCH, S2 1368 83

DISCH, S3 1679 124


DISCH, OET 749

DISCH, S1 1190 11

DISCH, S2 1524 43

DISCH, S3 970 -9


DISCH, OET 1068

DISCH, S1 1259 3

DISCH, S2 2111 73

DISCH, S3 2667 119


DISCH, OET 12185

1

2

4

© 2002 By Default!


Slide 61

Number of samples

➢Two or more samples to be taken

➢ If more than one BW source, than at least one sample taken from each BW source

Reasoning:

➢Average of two random samples shown to be representative, but more samples may be taken; and

➢As sample needs to be representative of whole discharge, if BW is loaded in different areas, BW water from each area needs to be sampled.

© 2002 By Default!


Slide 62

HOW MUCH VOLUME TO SAMPLE?

© 2002 By Default!


Slide 63

Sampled volume

STATISTICAL

representativeness

BIOLOGICAL

representativeness

lower higher

➢ missing diversity

➢ higher patchiness

representative

lower higher

➢ low confidence

e.g., 3x300-

500 litres

e.g., ~5.000 m3

empty e.g. 10.000 m3

➢ high confidence

➢ more stress

➢ organisms die

representative

© 2002 By Default!


Slide 64

➢ “BIOLOGICAL” representativeness is crucial!

➢ Statistical representativeness also to be considered

© 2002 By Default!


Slide 65

Sampled quantity

➢For organisms > 50; 300 – 500 litres of sample filtered and concentrated to ca. 5 litres fortransport / 100 ml for analysis

➢For organisms < 50 and > 10; 5 – 6 litres of continuous drip sample during sampling, subsample of ca. 100 ml for transport

➢For bacteria; 1 litre separated from thecontinuous drip sample

Reasoning:

➢Suggested methods and quantities showedbest results, concentrated sample easy tocarry

© 2002 By Default!


Slide 66

IN

SUMMARY

-

IN-LINE

Organism

group

Sampling

point

Equipment Water volume

[litre]

Number of samples

> 50

micrometres

In-line Plankton

net

300 – 500 in each

sequence

2 (or more) sequential

samples of ca. 10 minute

duration each, avoiding the

very beginning and very end

of the tank discharge event

< 50 and >

10

micrometres

In-line Sampling jar

and bucket

5 – 6 in each

sequence

2 (or more) continuous drip

sequential samples collected

at the same time as for

organism group > 50

micrometres

Indicator

microbes

In-line Bucket,

sampling jar

and bottle

1 in each

sequence

2 (or more) continuous drip

sequential samples sub-

sampled from the bucket

© 2002 By Default!


Slide 67

IN-TANK

SAMPLING GEAR

AND

(NO) SAMPLING POINT

ARRANGEMENTS

© 2002 By Default!


Slide 68

© 2002 By Default!


Slide 69

© 2002 By Default!


Slide 70

© 2002 By Default!


Slide 71

AVAILABILITY AND ACCESS OF SAMPLING POINTS

➢ sounding pipes should be on all tanks, but this is not the case

➢ air-vents are on all tanks, but is difficult to remove thecover

➢ manholes are on all tanks, but very frequently are not accessible because of cargo operation, cargo loadedon top, rusty bolts…

➢ tank hatches on few vessels, mainly tankers

© 2002 By Default!


Slide 72

IN-LINE

SAMPLING GEAR

AND

SAMPLING POINT ARRANGEMENTS

© 2002 By Default!


Slide 73

© 2002 By Default!


Slide 74

© 2002 By Default!


Slide 75

FLOW

METER

INFLOW

OUTFLOW

~ 2-2.5 m

~ 1m

© 2002 By Default!


Slide 76

© 2002 By Default!


Slide 77

PERSONAL PROTECTIVE

EQUIPMENT (PPE) IS

VERY IMPORTANT!

© 2002 By Default!


Slide 78

➢ chosen according to the sampling point

and local regulations

➢ avoid affecting the sample

➢ avoid infection

© 2002 By Default!


Slide 79

➢ BWS for CME is a complex process

➢ Different approaches and tools are available

➢ Organisms are patchy distributed in tanks what influences BWS results

➢ Very important is to chose the right approach and tools for the purpose of sampling (D-1, D-2, indicative, detailed)

➢ Methods and tools for BWS CME exist, Type Approval methods may be used

➢ New methods and tools may be developing

➢ New methods and tools need to be tested and compared first before use for CME

CONCLUSIONS

© 2002 By Default!


Slide 80

LITERATURE

© 2002 By Default!


Slide 81

LITERATURE

➢ Gollasch, S. & David, M. 2009. Results of an onboard

ballast water sampling study and initial considerations how to

take representative samples for compliance control with the D-

2 Standard of the Ballast Water Management Convention.

Report of research study for BSH, Hamburg, Germany, 11 pp.

➢ Gollasch, S. & David, M. 2010. Testing Sample

Representativeness of a Ballast Water Discharge and

developing methods for Indicative Analysis. Final report of

research study undertaken for the European Maritime Safety

Agency (EMSA), Lisbon, Portugal, 124 pp.

➢ Gollasch, S. & David, M. 2013. Recommendations for

Representative Ballast Water Sampling. Final report of

research study for BSH, Hamburg, Germany. 28 pp.

© 2002 By Default!


Slide 82

SUMMARY OF STUDY TEST VOYAGES + SHIPBOARD BWMS TESTS EXPERIENCE:

Report containing recommendations based on all studies andshipboard tests experience:

➢David, M. 2013. Ballast water sampling for compliance monitoring - Ratification of the Ballast Water Management Convention. Final report of research study for WWF International. Project number 10000675 - PO1368. 66 pp.

http://www.wwf.de/fileadmin/fm-wwf/Publikationen-PDF/Study_Silent_Invasion.pdf

http://www.wwf.de/fileadmin/fm-wwf/Publikationen-PDF/Study_Silent_Invasion.pdf

© 2002 By Default!


Slide 83













© 2002 By Default!


Slide 84

ACKNOWLEDGEMENTS

© 2002 By Default!


Slide 85

Ballast Water Management System for

Adriatic Sea Protection – BALMAS

http://www.balmas.eu/

ACKNOWLEDGEMENTS


© 2002 By Default!


Slide 86







© 2002 By Default!


Slide 87

THANK YOU

FOR THE

ATTENTION!

QUESTIONS

?

© 2002 By Default!


Slide 89

© 2002 By Default!


Slide 1

© 2002 By Default!


Slide 2

➢Background for targeting vessels and decision support system(DSS)

➢Risk assessment for targeting vessels for CME

➢CME DSS for targeting vessels

➢Tank selection criteria

➢ Conclusions

OUTLINE

© 2002 By Default!


Slide 3

➢ if there is no system to select vessels for BWM purpose inspection, the “critical” / “high risk” vessels would not be checked and may not be compliant, and instead “low risk” vessels couldbe checked

➢ level of risk posed by ballast water to bedischarged is a very important triggering element

➢ BWM Convention has some triggering elements in Article 9

➢ other triggering elements, e.g., information about non-compliance from previous port

➢ DSS may support PSC decision-making in this complex process

BACKGROUND

© 2002 By Default!


Slide 4

DSS to support CME

RA DSS BWM

Required

data input

CME

feedback

-

corrections

© 2002 By Default!


Slide 5

RA for TARGETING

VESSELS FOR CME

LEVEL OF RISK POSED BY BALLAST

WATER TO BE DISCHARGED?

© 2002 By Default!


Slide 6

MAIN GOALS:

➢ RA for each vessel arrival

➢ identify high risk vessels to enable CMEmeasures

➢ identify low risk vessels may lessen CMEmeasures

RA for BWM DSS

NEED FOR INFORMATION:

➢ ballast water to be discharged (donorport/area?), ballast water reporting needed

➢ salinity and presence of harmful aquatic organisms and pathogens (HAOP) in thedonor port

© 2002 By Default!


Slide 7

➢ data reliability (need for monitoring in ports)

➢ compare environmental compatibility (salinity)of the ballast water source area with the ballast water recipient area

➢ check the presence of HAOP in the ballast water donor area

➢ low risk assessed when very high environmental incompatibility and when no HAOP are present

➢ when no high environmental incompatibility and HAOP present, different levels of risk are triggered by different HAOP profiles, e.g., human pathogens, target species, harmful algae

RA principles

© 2002 By Default!


Slide 8

RA model

David, M., Gollasch, S. (eds.) 2015. Global Maritime

Transport and Ballast Water Management – Issues and

Solutions. Springer Science and Business media

© 2002 By Default!


Slide 9

DSS for TARGETING

VESSELS FOR CME

© 2002 By Default!


Slide 10

➢ vessel randomly selected under regular BWM verification process

➢ vessel selected for PSC inspection under separate process

➢ vessel not trustworthy

➢ ballast water discharge assessment(BWDA) dissagrees substanially withballast water reporting form (BWRF)

Triggering elements for

BWMC Tier1

(includes BWS)

© 2002 By Default!


Slide 11

➢ BWM Convention triggering elements (in port):

• no valid certificate

• condition of vessel or equipment don't

correspond with BWMS certificate

• master and that master and crew are not

familiar with BWM procedures/ not

implemented these

➢ PSA received notification that vessel is foundnon-compliant with D-2 standard in a previous port?

➢ Was any BW intended for discharge identified as extreme risk?

Triggering

elements for BWMC Tier2

Is it appropriate to conduct BWS for CME

under/after Tier2?

© 2002 By Default!


Slide 12

➢ OK as part of Tier 1

Discharge of ballast water not allowed when:

➢ no valid certificate

➢ condition of vessel or equipment don't correspond with BWMS certificate

➢ master and that master and crew are not familiar with BWM procedures/ not implemented these

➢ not when high or extreme risk is posed by theballast water to be discharged

When it is appropriate to

conduct BWS for CME

(detailed D2 test)?

© 2002 By Default!


Slide 13

CME model

David, M., Gollasch, S. (eds.) 2015. Global Maritime

Transport and Ballast Water Management – Issues and

Solutions. Springer Science and Business media

Was vessel

randomly selected under

regular BWM verification

process?

YES

Was vessel selected

for PSC inspection under

separate process?

IF

SUSPICIOUS

THEN

YES

Was vessel in

trustworthiness procedure

selected as "not

trustworthy"?

YES

Did BWDA model

disagree with BWRF? NO NO NO NO

YES

Vessel is

CLEAR TO PROCEED

until results from BWS analyses

are obtained.

PSC inscpection process as appropriate:

- interview master and BWM responsible

crew

- verify validity of BWMS certificate

- verify BWRB

- BWS for D-1 or D-2 (indicative and/or

detailed)

Proceed to

YES PSA

received notification

that vessel is found

non-compliant with D-2

standard in a

previous port?

NO

Was any BW

intended for discharge

identified as "extreme

risk"?

Proceed to

NO

YES

Clear grounds

that condition of vessel or

equipment don't

correspond with BWMS

certificate?

REGULAR

INSPECTION

Clear grounds

that master and crew

are not familiar with BWM

procedures and have not

implemented

these?

NO

Is vessel carrying

valid BWMS certificate?

YES

YES

NO

DETAILED

INSPECTION

PSC inscpection process as appropriate:

- interview master and BWM responsible

crew

- inspection of all needed documents and

log books

- inspection of vessel (BWMS)

- indicative BWS (without BW discharge)

YES

YESProceed to

YES

BWS was

conducted and results

confirmed

non-compliance?

Vessel

SHALL NOT DISCHARGE BW

until compliance

is confirmed.

NO

YES

NO

PSA DECISION 6:

Is vessel selected for

CME process?

NO

© 2002 By Default!


Slide 14

TANK SELECTION

WHICH TANK TO SAMPLE (FIRST)?

© 2002 By Default!


Slide 15

Such an assessment may consider

the following elements, but may not

be limited to:

➢the environmental compatibility of the ballast water donor area with the ballast water recipient area

➢the potential presence of HAOP in the ballast water donor area

➢if appropriate, the presence of target species in the ballast water donor area, and

➢the in-tank holding time

Tank selection criteria

© 2002 By Default!


Slide 16

➢ the higher the risk identified, the higher the priority

➢ give priority to tank(s) from anotherbiogeografical area with with higher environmental compatibility with the discharge area

➢ give priority to the tanks with ballast water origin area where HAOP or target species are present

➢ give priority to tanks with shorter in-tank holding time

➢ give priority to tanks to be dischargedfirst

Some priority principles

© 2002 By Default!


Slide 17

➢ for in-tank sampling accesibility, the ease of the sampling access point may be used as an additional criterion to identify the tank to be sampled, considering also that ballast water in some tanks may not be accessible at all for in-tank sampling

➢ for in-line sampling the tank(s) which is(are) currently being discharged when PSC comes on board may be sampled first

• to enable stop of discharge if needed

• not to wait until the »targeted tank« is ready

to be discharged

Some additional

considerations

© 2002 By Default!


Slide 18

➢A system is needed for selection of vessels and tanks for CME for the purpose of the BWM Convention

➢Important to have a regional PSC approach implemented (MoU)

➢Risk assessment is an important tool to support the CME process

➢DSS for targeting vessels eases the PSC decision-making process

CONCLUSIONS

© 2002 By Default!


Slide 19

Vectors of Change in Oceans and

Seas Marine Life, Impact on

Economic Sectors - VECTORS

http://www.marine-vectors.eu/

ACKNOWLEDGEMENT

http://www.marine-vectors.eu/

© 2002 By Default!


Slide 20

ACKNOWLEDGEMENT

Ballast Water Management System

for Adriatic Sea Protection –

BALMAS



© 2002 By Default!


Slide 21













© 2002 By Default!


Slide 22







THANK YOU

FOR THE ATTENTION!

QUESTIONS

?

© 2002 By Default!


Slide 24

• Recommendations for

– Sample labelling

– Sample transport to the laboratory

– Chain of custody

– Sample handling in the laboratory

Outline

• Document sampling event details

• Label bottle (not lid)

– Sample ID code

• Date, start/end time

– D-2 organism group

– Sampling point

– Water volume sampled

– Preservative (if used)

• Secure label with transparent

tape or put samples separately

in bags

Sample Labelling

• Transport sample to lab as soon as possible

• Shipment may also be needed when

analysis requires special lab (Cholera)

– Styrofoam box

– Cooling / heating elements if needed

• Ship sample in controlled conditions

• Document temperature with

temperature loggers

• Sample shipment temperature

– Plankton - slightly cooler than sampling temperature

– Abiotic – according to standard method (ISO etc.)

Sample Transfer to Laboratory

• Documentation of sample shipment

• Record of

– Sampling event details

– Purpose of shipment

(which analysis is needed)

– Sample types

– Number of samples

• All involved to sign

• Cholera = C samples

Chain of Custody

• After sample arrived, if not yet done on board

– Fill 1 L bottle for bacteria analysis

– Fill 2 x 100 ml bottles with phytoplankton

sample and add preservative to one

– Empty cod-end into filter

(30 µm mesh)

– Wash filter mesh content into jar

– Fill wash bottle with filtered

water

– Bring wash bottle and jar

to microscopist

Sample Handling

• Label samples that they can clearly be identified

and are not mixed

• Transport samples to laboratory as soon as

possible

• Document sample transport conditions and

involved persons (chain of custody)

• Handle sample for all D-2 organism groups

• Very well clean all gear to avoid organism

„contamination“ with the next samples

Summary

• Sampling purpose

• D-1 standard

• Possible methods for compliance checks

Outline

• Compliance with D-1

Ballast Water Exchange (BWE)

– 200 m depth, 200 nm from nearest land

– 200 m depth, 50 nm from nearest land

– Designated areas

Sampling Purpose

• Tracers of human activity

– Presence of e.g. Nitrogen or Phosphorous may

indicate nearshore BWE (river run-off in urban areas),

colored dissolved organic matter (CDOM)

• Coastal species

– Harpacticoid copepods, barnacles

• Sediment

• High sediment load may indicate near-shore BWE, but

re-suspension from tank bottom occurs

• Salinity

If salinity is below 30 psu it is unlikely that it was

exchanged at sea

D-1 (Ballast Water Exchange)

D-1 (Ballast Water Exchange)

• Conductivity sensor

– easy to calibrate (with freshwater)

– portable

– dip-stick method

• Refractometer

– easy to calibrate

(with freshwater)

– portable (pocket-size)

– a few drops of water are sufficient

Tools for D-1 Compliance Checks

• Methods exist to proof D-1compliance

• We recommend to measure water salinity, but

– BWE in coastal/port areas in high-salinity

environments (above 30 psu) will results in

compliance but water does not originate from mid-

ocean (non-compliance with D-1); or

– BWE in designated ballast water exchange areas

(within 50 nm from nearest land) may result in less

salinity than the mid-ocean water. Here the BWE

would be compliant, but salinity measurements alone

would show non-compliance

• Origin of BWE must be known

Conclusions

Stephan Gollasch

GoConsult, Hamburg, Germany

[email protected]

www.gollaschconsulting.de

Matej David

Dr. Matej David Consult d.o.o.

Korte, Slovenia

[email protected]

www.davidconsult.eu

Contacts

><((((°> GoConsult

• Sampling purpose

• D-2 standard

• Ballast water biology

• Indicative vs detailed analysis

• Organisms detection methods

• Viability

• Minimum dimension

Outline

• Compliance with D-2 (Ballast Water Performance Standard)

– <10 viable organisms/m³ >50 µm

– <10 viable organisms/ml <50 & > 10µm

– Indicator microbes

• Escherichia coli <250 cfu in 100 ml

• Enterococci <100 cfu in 100 ml

• Vibrio cholerae <1 cfu in 100 ml or in 1 gr ww

zooplankton

(Cfu=Colony forming units)

Sampling Purpose

Ballast Water Biology

• Almost all types

of organisms

• Dominant are

crustaceans,

mollusks, worms

and phytoplankton

• 15 cm long fishes

found in tanks

• Harmful algae

• Pathogens

Ballast Water Biology

• Indicative analysis

– A „quick and dirty“ check

for gross exceedance;

e.g., 100s organisms =

non-compliance

• Detailed analysis

– A detailed analysis;

e.g., 10 organisms =

non-compliance

Two Different Approaches

• EMSA Study

2010

• Interreg IV B

Ballast Water

Opportunity

2012

• BALMAS

2014

• BSH ReBaT

Project 2016

www.bsh.de

Summaries of Organism Detection

Methods

• An evaluation of the following

was done:

– Accuracy

– Reliability

– Time to a result

– Expertise

– Portability

– Costs

– Use on board

or in lab

Possible analysis methods

Indicative Analysis Methods

D-2 organism groups

-Viable organisms less than 50 and greater than or equal to 10 micrometers in minimum dimension

12 methods considered

-Viable organisms greater than or equal to 50 micrometers in minimum dimension


-Methods for bacteria analysis


Organisms <50 and ≥10 µm in minimum dimension

Presence/absence (biomass, no counts)

-e.g. DNA, ATP, Chlorophyll a methods deliver results in less than 60 minutes

Viability and estimated counts

-Pulse-amplitude modulated (PAM) fluorometry, portable

-Flow cameras or flow cytometry (< 60 minutes, not portable, viability stain needed)


Organisms <50 and ≥10 µm in minimum dimension

Best compromise:

Pulse-Amplitude Modulated (PAM) fluorometry

-portable, easy to use, low expertise needed

-viability in less than 10 minutes onboard (by PSC)


-PAM measures phytoplankton biomass and viability

-No direct counts („estimated numbers“)

-Clear relation of biomass and viability measurements with organism numbers

-Suitable tool to show that D-2 was not met

-Detection limit is 1 org / ml (calculated) 0

50

100

150

200

250

300

0 0,1 0,2 0,3 0,4 0,5 0,6 0,7 0,8

Organism viability

Nu

mb

er

of

sm

all

er

org

an

ism

s /

ml Total number of smaller

organisms

Pulse-Amplitude Modulated Fluorometry (PAM)

Indicative Analysis Methods Organisms ≥50 µm in minimum dimension

Presence/absence methods (no counts)

-e.g. DNA, ATP methods deliver results in less than 60 minutes

Counts (no viability)

-Hand-held flow cameras results in less than 30 minutes

Best compromise: Stereomicroscope (counts & viability) in laboratory

-results in less than 20 minutes

-portable, easy to use, but high expertise needed


D-2 Bacteria

Presence/absence methods (no cfu and/or counts)

-e.g. DNA, ATP methods deliver results in less than 60 minutes

All methods to determine cfu require incubation time of >6 hours

-Start with one method to evaluate one organism group in D-2

-Should this show presence or high numbers, take result as indication of a failure to meet D-2

-Should this show absence or low numbers, continue with second (and third) D-2 organism group to confirm results

-The easiest to start with may be the analysis for phytoplankton (Pulse-Amplitude Modulated fluorometry) on board and if needed followed by zooplankton (stereomicroscope) in laboratory


Suggestions

Indicative Analysis Approach

Organisms <50 - ≥10 µm

(PAM) on board

Organisms ≥50 µm

(stereomicroscope) lab

D-2 met

D-2 met

D-2 compliance

action

D-2 non-compliance

action

D-2 not met

D-2 not met

-Ballast Water Monitoring A/S, Denmark. Fluorescence

-Ballast Water Checker, China. PAM fluorescence

-BallastWISE, Denmark. Movements and fluorescence

-P. Counter, Republic of Korea. Fluorescence and image

processing

-ChemChrome V6 (CV6), France. Stain

-Ballast Eye, Japan. FDA

-Integrated Ballast Testing,

Spain. Image cytometry

-B-QUA, Canada. ATP

-Aquatools, France. ATP

Indicative Analysis Methods Recent

Developments

• Work according to good laboratory practice standards

• Most accurate analysis technologies should be used

• At best consider living samples

• Keep time between sample taking and sample analysis

as short as possible

• Unlikely to have one detailed sample processing

standard world-wide

Detailed Analysis

• IMO Code G8 - Definition: “Viable organisms means

organisms that have the ability to successfully generate

new individuals in order to reproduce the species.”

• How to proof this?

• Imagine

– You only have one individual and no breeding mate

– You only have females in your sample

– You have species which reproduce once a year

Viability Problem

• IMO Code G8 – Sample Analysis: “Viability may be

established by assessing the presence of one or more

essential characteristics of life, such as structural

integrity, metabolism, reproduction, motility or response

to stimuli”

– structural integrity = complete organisms plus

– metabolism = organ activity, heartbeats or

– reproduction = new generation or

– motility = movements or

– response to stimuli = poking

Viability Solution

• Phytoplankton

– viability stain Fluorescein DiAcetate (FDA)

– 5-ChloroMethylFluorescein DiAcetate (CMFDA)

• Zooplankton

– initiate movement with light under stereomicroscope

– gentle poking with a needle

• Bacteria

– colony forming means viable

Viability

• Stains living phytoplankton

– Cell-permeant esterase substrate documents

enzymatic activity (needed to activate its

fluorescence) and cell-membrane integrity (needed

for retention of fluorescent product in cell)

– Problem with UV treated

water

Viability Stain Fluorescein

DiAcetate (FDA)

• Look at the organisms from all axes

• Select the smallest axes

• Measure the smallest dimension

• Not always possible!

• Use smallest visible axis

Minimum Dimension

• IMO G2: The "Minimum Dimension" of an organism

based upon the dimensions of that organism's body,

ignoring, e.g., spines, flagellae or antenna.

• The minimum dimension

should be the smallest part of

the "body", i.e. the smallest

dimension between main

body surfaces of an individual

when looked at from all

perspectives.

Minimum Dimension Issues

• Never fully accurate when using filters to separate D-2

organism size classes

• Problems “organism damage” and “minimum dimension”

– Organisms with long spines are

caught in filter although their

minimum dimension was smaller

– Colonies caught although individual

may be smaller than filter mesh

– Spaghetti in pasta drainer never

slip through although the would fit

through the holes

Problems with Filtration

www.kochwiki.org

• Several algae form colonies or chains

• What if a colony or chain is above 50 µm in minimum

dimension and the single individual below 50 µm?

• Measure and count the single individual

– Because D-2 refers to viable organisms and it is the

individual being viable and not the colony

Colony vs. Single Organisms

200 µm

40 µm

• Suggested methods

– Organisms <50 and ≥10 µm

• Phytoplankton - Epifluorescence microscope

• Zooplankton – Microscope

– Organisms ≥50 µm

• Phytoplankton - Microscope

• Zooplankton – Stereomicroscope

– Bacteria

• Selective media

• Consider routine bacteria analysis methods

(drinking / bathing water quality standards)

Detailed Analysis

D-2 organism <50 and ≥10 µm

-Epifluorescence microscopy

-Fluorescein diacetate (FDA)

-1 ml Sedgewick-Rafter chamber with grid of 1000 squares

-Grid lines = 10-15 µm

-Count vertical transects

Sedgwick-Rafter

chamber

Detailed Analysis Methods

D-2 organisms ≥ 50 µm

-Microscopic analysis

-Stains don´t work 100%

-Living dead judgement by a scientist

-50 µm beads or mesh for size

Stephan

counting

chamber


D-2 methods for bacteria

-colony forming units to identify

-not all bacteria form colonies

-incubation needed

-selective media


-Consider to equip a van with organism detection technology

-Drive from vessel to vessel in a port

-Send sampling team onboard and deliver the samples as soon as possible to van for analysis

-In this scenario the organism detection tools would not need to be carried onboard

-Sampling team “only” to board the vessel, no need to bring organism detection team on board as well

Analysis Methods

Suggestions

Analysis Methods, Summary Organism Indicative

≥ 50 µm Stereomicroscope

(in laboratory)

< 50 and ≥ 10 µm PAM

(on board method)

Bacteria Developing

Organism Detailed

≥ 50 µm Stereomicroscope

(needle for poking)

< 50 and ≥ 10 µm Epifluorescence

microscope with

CM/FDA stain

Bacteria Selective media

• Methods exist to proof D-2 compliance

• New methods need to be validated

• Harmonized approach needed, not that one vessel is

compliant in one port, but not in another because of

different sample processing methods used

• An indicative sample analysis may be followed by a

detailed analysis of the same sample

Conclusions

Stephan Gollasch

GoConsult, Hamburg, Germany

[email protected]

www.gollaschconsulting.de

Matej David

Dr. Matej David Consult d.o.o.

Korte, Slovenia

[email protected]

www.davidconsult.eu

Contacts

><((((°> GoConsult

1

QUESTIONNAIRE

1. Are you planning future Ballast Water testing in your country?

Latvia: Latvian Institute of Aquatic Ecology will be responsible for ballast water sampling, analysis

and environmental risk assessment for the granting of exemptions.

Estonia: Ballast Water testing will be conducted for the first time in 2019.

Finland: Not at the moment. Finland has conducted a sampling study, where four different

indicative sampling devices have been tested. The results were compared to detailed analysis. The

report will be published soon.

Denmark: Denmark is currently planning how to implement ballast water sampling and analysis

according to IMO regulations.

Germany: Yes, Germany is implementing a project for ballast water sampling and analysis. The aim

of this project is to find devices that deliver representative results and are easy to use during ship

surveys conducted by PSCOs or Water Police officers.

2. Are there already concrete plans?

Latvia: Based on the Latvian Law of “International convention for the control and management of Ships' ballast water and sediments, 2004”, entry into force on 04.07.2018, there are responsible institutions nominated, but there is no concrete plan for coordinated actions. Estonia: Planned is taking samples for microbes (incubation + genetics) and check the water in tank for compliance with D-1. Finland: No concrete plans yet. However, Finland plans to purchase indicative analysis devices in the near future. Denmark: So far only desktop draft plans – but planning is in progress. Germany: Yes, the first training on indicative analysis for Water Police officers will start in July. In August there will be tests on different sampling methods. After August tests of different methods for sampling and analysis on board during compliance control will be performed.

3. Which institutions/administrations are involved?

Latvia: Since the Law of “International convention for the control and management of Ships' ballast water and sediments, 2004” is in force, the Ministry of Transport and the Ministry of Environmental Protection and Regional Development coordinate the fulfillment of obligations under the Convention. The other responsible institutions are: Ministry of Education and Science,

2

The State Environmental Service of the Republic of Latvia, Maritime administration of Latvia, Latvian Institute of Aquatic Ecology and coast Guard Service of Latvian Naval Forces. Estonia: The Ministry of the Environment is funding the applied research project on testing the microbiology in ballast water. The Marine Ecology Lab of the Department of Marine Systems (TalTech) and microbiologists from the University of Tartu will be involved. Finland: Finnish Transport and Communications Agency is responsible for PSC inspection in Finland. Consultative help can be asked from Finnish Environment Institute (SYKE). Denmark: The Danish Maritime Authority in relation to PSC (Paris MOU: certificate, journal and plan) and The Danish Environmental Protection Agency in relation to BW sampling and analysis and the subsequent verification of the results for compliance or non-compliance. It is expected that PSC will notify EPA if there is suspicion of non-compliance. Germany: The Federal Maritime and Hydrographic Agency, the Water Police departments of different German Federal States as well as the German Social Accident Insurance Institution for Commercial Transport, Postal Logistics and Telecommunication (Ship Safety Division, BG Verkehr).

4. Do you need additional information/consulting to do so?

Latvia: At the moment further co-operation within the national institutions in the country is necessary, as there are no clear schemes of coordinated actions. Estonia: Might need the informational support e.g. on sampling or analyzing the samples/results. Finland: An International Standard for representative sampling is necessary to conduct future inspections. Denmark: For now no, but perhaps in a year or so from now a follow-up-workshop or a just a simpler questionnaire could be beneficial to do for all involved parties to share experiences and knowledge on the logistical and practical challenges involved in CME. Especially regarding the costs involved for sampling and analysis as well as man hours involved. Germany: No, not for now.

5. Are there co-operations with other countries concerning testing?

Latvia: At the moment there are no co-operations with other countries concerning testing. Estonia: At the moment there is no foreign partner concerning testing, but Estonia is open for collaboration and exchange of the experience. Finland: No Denmark: So far not. However, Denmark is of course willing to share experiences and to collaborate with other countries if relevant to align testing procedures.

3

Germany: No, there are no co-operations but an exchange of information.

6. From your experience and perspective: What are the most important aspects which have to be clarified, problems which have to be solved regarding BWMC compliance monitoring?

Latvia: Co-operation within the responsible and involved authorities regarding vessel inspection, sample collection, reporting. Granting of exemptions becomes more and more urgent issue within the next few months. Estonia: The problem and its solving have to be internationally accepted and monitoring of the ballast water have to be developed, in collaboration with different stakeholders and research institutions, approved methods and standardized techniques (e.g. instruments for testing the compliance) for routinely held monitoring in the different regional seas (e.g. Baltic Sea). Finland: - Taking representative sample for detecting the ≥ 50 µm size fraction is difficult in practice. - Availability of laboratories for detailed analysis is unclear. - At the moment we do not have laboratories in Finland, which could analyze toxicogenic Vibrio cholera (01 and 0139) from sea water sample. Denmark: Equipment and methods for BW sampling and the required resource competency for BW sampling and indicative analysis are our main bottlenecks. Subsequently, what resources are available (need to be available) to carry out the whole testing procedure? In more detail: How it is possible to obtain an effective and sound testing procedure including BW sampling and analysis which must be done by experienced and competent authority/technicians. Furthermore, they need to be able to move out to different parts of the country with short notice (also outside normal office working hours) with the relevant and required equipment for sampling and analysis of the samples taken (keeping in mind that only two of the three size groups can currently be verified with existing methods). And finally, how to go through all this without delaying the vessel. Germany: For the German project on the experience building phase the equipment for sampling and analysis needs to be light in weight and easy to use while providing representative and reliable data within a short additional time during the regular surveys by water way police officers or PSCOs.

1

PARTICIPANTS Estonia Toomas Mironov Estonian Environmental

Research Centre

Ivan Kuprijanov Marine Systems Institute at

Tallinn University of Technology

Finland Jukka Huhtanen Finnish Transport and

Communications Agency

Juha Suomalainen Finnish Transport and

Communications Agency

Germany Mike Meklenburg BG Verkehr, Ship Safety Division

Sebastian Seibel Water Police Mecklenburg-

Vorpommern

Octavio Marin Maritime and Hydrographic

Agency

Lithuania Greta Sriebalienė Marine Research Institute,

Klaipėda University

Paulius Petrošius Environmental Protection

Agency, marine monitoring

department

Eglė Šupinienė Environmental Protection

Agency, marine monitoring

department

Poland Grzegorz Siemiński Maritime Office in Gdynia

Piotr Prokop Maritime Office in Szczecin

Denmark Kim Lundgreen Danish EPA

Latvia Ieva Barda Latvian Institute of Aquatic

Ecology

Ieva Putna-Nimane Latvian Institute of Aquatic

Ecology

Ballast Water Sampling and Analysis Workshop - Balticcomplete

Documents