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RESEARCH STUDY
Ballast water sampling for compliance monitoring -
Ratification of the Ballast Water Management
Convention
(Project number 10000675 - PO1368)
FINAL REPORT
I
Title:
Ratification of the Ballast Water Management Convention
(project number 10000675 - PO1368)
WWF International
Funding organisation:
Avenue du Mont-Blanc
1196 Gland
Switzerland
WWF International by WWF Sweden
Funds provided by:
Prof. Dr. Matej David,
Lead Researcher:
Dr. Matej David Consult s.p.
Korte 13 e
6310 Izola
Slovenia
Korte, Slovenia, March, 2013
II
III
Citation and disclaimer
This report should be quoted as follows:
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.
The contents and views contained in this report are those of the author, and do not
necessarily represent those of the WWF International.
IV
ABBREVIATIONS
BLG Sub-Committee on Bulk Liquids and Gases (of IMO)
BWM Ballast Water Management
BWM Convention International Convention for the Control and Management of Ship’s
Ballast Water and Sediments
BWMS Ballast Water Management System
BWS Ballast Water Sampling
D-1 Ballast Water Exchange Standard, Regulation D-1 of the BWM
Convention
D-2 Ballast Water Performance Standard, Regulation D-2 of the BWM
Convention
G2 IMO Guidelines for Ballast Water Sampling
G8 IMO Guidelines for the Approval of Ballast Water Management
Systems
ICES International Council for the Exploration of the Sea
IMO International Maritime Organization
IOC Intergovernmental Oceanographic Commission (of UNESCO)
MEPC Marine Environment Protection Committee (of IMO)
PSC Port State Control
David M., Ballast water sampling for compliance monitoring, Final report. WWF Project
V
TABLE OF CONTENTS
ABBREVIATIONS ........................................................................................................... 4
EXECUTIVE SUMMARY ................................................................................................ 1
1 INTRODUCTION ....................................................................................................... 5
2 OBJECTIVES ............................................................................................................. 7
3 MATERIALS AND METHODS ............................................................................... 8
4 BALLAST WATER SAMPLING GUIDANCE .................................................... 10
4.1 COMPLIANCE CONTROL SAMPLING .................................................................... 10
4.2 SAMPLING METHODS ACCORDING TO THE ACCESS POINT ................................. 11
4.3 TANK SELECTION – WHICH TANK TO SAMPLE (FIRST)? ..................................... 12
4.4 SAMPLING FOR COMPLIANCE WITH THE D-1 STANDARD ................................... 13
4.4.1 In-tank D-1 standard sampling ................................................................. 16
4.4.2 In- line D-1 standard sampling ................................................................. 18
4.5 SAMPLING FOR COMPLIANCE WITH THE D-2 STANDARD ................................... 19
4.6 INDICATIVE SAMPLING ....................................................................................... 20
4.6.1 In-tank indicative sampling ...................................................................... 21
4.6.2 In- line indicative sampling ...................................................................... 24
4.7 DETAILED SAMPLING FOR COMPLIANCE WITH THE D-2 STANDARD ................. 26
4.7.1 In-tank D-2 sampling ................................................................................ 26
4.7.2 In- line D-2 standard sampling ................................................................. 26
4.7.3 Recommendations for a ballast water sampling protocol that is
representative of the whole discharge ................................................................. 28
4.7.4 Sampling logistics feasibility ................................................................... 30
4.8 SAMPLING EQUIPMENT AND SAMPLING POINT ARRANGEMENTS ....................... 31
4.8.1 Sampling equipment ................................................................................. 31
4.8.2 In-tank sampling arrangements on vessels ............................................... 41
4.8.3 In- line sampling arrangements on vessels ............................................... 43
4.8.4 Sampling point .......................................................................................... 46
4.9 SAMPLES HANDLING ........................................................................................... 48
4.9.1 Samples labelling ...................................................................................... 48
4.9.2 Samples transport ..................................................................................... 49
4.9.3 Chain of custody ....................................................................................... 49
5 DISCUSSION ............................................................................................................ 50
David M., Ballast water sampling for compliance monitoring, Final report. WWF Project
VI
6 CONCLUSIONS AND RECOMMENDATIONS .................................................. 52
7 ACKNOWLEDGEMENTS ..................................................................................... 55
LIST OF FIGURES ........................................................................................................ 56
LIST OF TABLES ......................................................................................................... 57
REFERENCES .............................................................................................................. 58
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EXECUTIVE SUMMARY
To proof compliance with ballast water management standards the IMO Ballast Water
Working Group developed a sampling guideline, i.e., Guidelines for Ballast Water Sampling
(G2 Guidelines), adopted in 2008. It was noted that G2 lacks technical details regarding
compliance control sampling with the standards as outlined in Regulations D-1 and D-2 of the
Ballast Water Management Convention. Consequently additional documents were prepared
by BLG to address this:
• an aide-memoire on the analysis of samples developed at BLG13 (March 2009). BLG
agreed that additional guidance on sampling procedures could not yet be developed
because of the lack of relevant study results (BLG13/WP.5, Annex 1). This aide-
memoire is entitled “Initiate Development of an IMO Circular to provide Ballast
Water Sampling Analysis Protocols and to give Advice on Uniform Application of the
Protocols” and is limited to types of sample analysis, including recommendations for
indicative and detailed sample processing.
• preparational documents of the Ballast Water Sampling Circular (e.g.
BLG15/5/1, BLG15/5/4-6). This IMO Circular was aimed to give ballast water
sampling and analysis guidance, including sample representativeness and sample
analysis protocols.
• the report of the Ballast Water and Biofouling Working Group from the last BLG
meeting (BLG17, February 2013) contains the draft Ballast Water Sampling
Circular (i.e., Guidance to Ballast Water Sampling and Analysis for Trial Use in
Accordance with the BWM Convention and Guidelines G2). The draft BWS circular
was prepared for consideration at MEPC65 and is made up of two parts: (a) a
discussion of the principles of sampling, accompanied by a list of recommended
methods and approaches for analysis and sampling protocols available for compliance
testing to the D-1 and D-2 standards; and (b) background information on sampling and
analysis methodologies and approaches. However, it is stated in the document that
methods for representative sampling are still developing (BLG17/WP.4) so that no
details are included regarding the number of samples to take or on their volume.
In view of this, WWF International noted the need to provide additional information
which may be considered during the further development of the BWS circular and supported
this study with the aim of overcoming sampling uncertainties in order to avoid any further
delay in the ratification of the Convention. Consequently, this detailed report was prepared to
address ballast water sampling recommendations for sampling for compliance with the D-
1 and D-2 standards, indicative and detailed sampling, in-tank and in-line sampling. In
addition, recommendations are given on sampling equipment, sampling arrangements on
the vessel, and sample handling and transport.
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For sampling for compliance with the D-1 standard in-tank or in-line samples may be
taken to either proof the presence of coastal biota or check the water parameters. Salinity
values verify if the water has been exchanged according to the BWM Convention
requirements, i.e. in cases where the salinity is low (e.g., below 30 psu) it can be assumed that
the ballast water originates from coastal areas with freshwater influence, i.e. it was not
exchanged as ocean water (i.e., outside 50 or 200 nautical miles from nearest land and at
water depths higher than 200 metres) as it would have a higher salinity. For this purpose,
small quantities of ballast water may directly be sampled from the tank via sounding pipes or
manholes prior to its discharge. However, this is a feasible option only when the inspected
vessel has loaded ballast in a low salinity or freshwater port. A problem with the in-line
sampling is that a discharge to sea may occur during the sampling and in case of non-
compliance the non-complaint water would enter the recipient environment during the
sampling event.
The G2 Guidelines do not address explicitly how an indicative sampling event would need to
be undertaken. Implicitly, an indicative analyses could be performed on a sample, or part of it,
taken during the detailed D-2 standard compliance control sampling process, or just on any
stand-alone sample.
For indicative in-tank sampling it was concluded that sampling for zooplankton via the
sounding pipes does not result in a representative sample of species in the tank as
comparisons of sounding pipe and manholes samples from the same tank found that net
sample delivery was more diverse biologically. Further, pumps used via open manholes
delivered more diverse samples than net samples, therefore pumps may also be considered
when sampling via manholes. A problem is that frequently manholes cannot be opened due to,
e.g., overlaying cargo or ongoing cargo operations in the area where the manhole is located,
and in these instances sounding pipe sampling might be the only solution to sample the ballast
water at all.
When applying indicative in-line (at discharge) sampling, it is recommended that one
sequential sample is taken using the same sampling methodology as for a detailed D-2
standard compliance test. However, having in mind that indicative sampling may also be used
to have an early indication of potentially non compliant ballast water, and consequently to
apply appropriate management measures, one key problem remains, and this is that
compliance or non-compliance can in this scenario only be proven while the ballast water is
being pumped overboard. Should, e.g., a risk assessment result in the identification of high
risk ballast water to be discharged the in-line sampling during discharge should be avoided,
but in-tank sampling should be undertaken to assess compliance.
To proof compliance with the D-2 standard, i.e., detailed sampling, being a numerical and
biological discharge standard, the samples should be taken from the ballast water discharge
line as near to the point of discharge as practicable, during ballast water discharge whenever
possible. Although in-line sampling seems most appropriate to assess compliance with a
discharge standard, in case the ballast water originates from a high risk area, i.e. an area with
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a known occurrence of target species, samples may be preferably taken from the ballast tank
prior discharge. This enables non-compliance actions before the water is pumped into the
recipient environment. The in-tank method may also be the only way to proof compliance
with the D-2 standard for tanks with direct discharges to sea.
In previous studies, which compared sequential samples with samples taken over the entire
discharge time of a ballast tank, the sequential trials, i.e., sampling over shorter sequences of
ca. 10 minutes each, showed different organism numbers in each sequence of one test
indicating the patchy organism distribution inside the ballast tank. This was observed during
all sampling events and for both organism groups studied. Hence, sampling during ballast
water discharge is biased by tank patchiness of organisms.
For the group of organisms greater than or equal to 50 micrometres in minimum
dimension the previously undertaken studies showed that samples taken over the entire
discharge time of a tank contained much lower living organism concentrations compared to
the living organism count in the sequences so that sequential sampling may deliver more
representative results.
It was further observed that sequential samples taken in the very beginning and end during a
ballast tank is emptied are unlikely to give representative results of the living organism
concentration because in these samples the organism count showed high variations, which
may result in under- or oversampling the organism concentration. Even when avoiding these
time windows the concentration of living organism still seems to be patchy so that it is
recommended to take at least two sequential samples in this time window avoiding the very
beginning and end during a ballast tank is emptied. The mean value of the living organism
concentration in these two samples may be taken to assume the real organism concentration.
Comparative studies have shown that sequential samples of approximately 10 minutes
duration which delivers a sample volume of 300 – 500 litres are suitable for detailed in-line
D-2 standard compliance tests. It is recommended that the water is concentrated during the
sampling process so that the water volume a PSC may need to transport to a laboratory for
subsequent analysis becomes a handy size. For this water concentration a plankton net should
be used which is placed in a bucket to the maximum level to ensure organism save sampling.
For the group of organisms less than 50 micrometres in minimum dimension and greater
than or equal to 10 micrometres in minimum dimension cell counts of sequences in
comparison to the samples taken over the entire time of a tank discharge showed similar or
lower living organism concentrations in sequential samples, which is in contrast to the larger
organism results.
The comparison of the phytoplankton cell concentration between the different sequences of
all tests showed that no clear trend can be identified during which time window a more
representative sample will be taken. Therefore it is recommended to take at least two
sequential samples of 5 - 6 litres volume during the discharge of a ballast water tank with
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avoiding taking a sample during the very beginning and end of the discharge time of a tank or
tanks. These samples should be taken over a 10 minute or less time interval, i.e., during
sampling for organisms greater than or equal to 50 micrometres in minimum dimension, as
continuous drip sample. The mean organism count in two or more samples may be seen as the
real living organism concentration in the ballast water.
For indicator microbes, it is recommended to follow the same sampling approach as for the
organisms less than 50 micrometres in minimum dimension and greater than or equal to 10
micrometres in minimum dimension, but that the water volume to be sampled should be
approximately 1 litre. The sample can be also subsampled from the water sampled for the
organisms less than 50 micrometres in minimum dimension and greater than or equal to 10
micrometres in minimum dimension collected in a bucket.
Sampling equipment for D-1 and D-2 standards sampling is available and includes plankton
nets, water samplers, pumps, buckets, sampling bottles etc, which can be employed as per the
sampling point appropriateness and availability on board.
In conclusion the ballast water sampling guidance presented in this report is based on results
and experience from earlier ballast water sampling studies and G8 type approval tests of
different ballast water management systems on ships voyages, also considering vessels
ballasting practices, practicability for PSC compliance monitoring, and the reliability of
results. These methods were developed and tested on board of commercial vessels and also
used for type approval of different BWMS, and were further also scientifically tested,
improved and validated during three studies on representative BWS for compliance
monitoring, conducted for the Federal Maritime and Hydrographic Agency, Hamburg,
Germany in 2009 and 2012, and for the European Maritime Safety Agency, Lisbon, Portugal
in 2010.
During BLG17 a trial period was suggested to test, further develop and scientifically validate
ballast water sampling methods contained in the report of the Working Group on the
Development of Guidelines and other Documents for Uniform Implementation of the 2004
BWM Convention (IMO 2013) for the purpose of compliance control. It is hoped that more
studies will be initiated in the near future to meet this objective. Results from this trial period
will likely be used to update the BWS circular.
This study has shown that there are BWS methods which were extensively used on board
vessels to test BWMS to proof compliance especially with the D-2 standard, and were
scientifically validated by additional tests and studies. These BWS methods have also shown
to be relatively simple, i.e., no special background education is needed for their application,
they are cost effective, i.e., there is no need for very expensive equipment, and there are no
high running costs, and they are generally applicable on all vessel types and in all geographic
regions. The sampling recommendations suggested in this study therefore may result in a
workable, equitable and pragmatic solution to support the entry into force of the BWM
Convention.
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1 INTRODUCTION
The International Convention for the Control and Management of Ship’s Ballast Water and
Sediments (BWM Convention) will enter into force 12 months after ratification by 30 States,
representing 35 per cent of the world merchant shipping tonnage. As of 06 March 2013, 36
States have ratified the BWM Convention, representing 29.07% of the world merchant
shipping tonnage (IMO, Status of conventions). However, despite being so close to
ratification, the economic situation and other issues arising from perceived uncertainties has
had a huge delay effect on flag states being willing to ratify the BWM Convention.
The BWM Convention includes Regulation D-2 Ballast Water Performance Standard (D-2
standard) which specifies water quality requirements, in the form of upper concentration
limits of living organisms in ballast water discharges.
The lack of ballast water sampling/testing standardisation to document compliance with the
D-2 standard is perceived as substantial uncertainty. The delay in the BWM Convention
ratification and consequently its entry into force may also be attributed to the need for the
finalisation of the ballast water sampling (BWS) guidance documents at IMO. These
documents are prepared as additional guidance documents to the G2 guidelines.
A conclusion of the discussions at recent BLG 17 and MEPC 64 meetings was that, even
when a certified BWMS is operated correctly, it still may fail the D-2 standard compliance
testing during a port State control (PSC) inspection as the G8 Guidelines are considered not
robust enough to ensure that a ballast water management system (BWMS) will perform well
enough to meet the D-2 standard in all water conditions. It was therefore agreed at MEPC 64
that BWS Guidance being developed for PSC compliance inspections should not use more
stringent testing parameters than those required in the G8 Guidelines. This means, that
compliance control may be workable, but the question remains if it is conducted to an
acceptable level of accuracy to proof concentrations of living organisms in ballast water
discharges?
To improve its robustness, the IMO BWS Guidance text was just updated and finalised at
BLG 17 in a form of a BWS draft circular, i.e., Guidance to Ballast Water Sampling and
Analysis for Trial Use in Accordance with the BWM Convention and Guidelines G2.
The IMO BWS Guidance also notes that improvements may be necessary when additional
knowledge and experience becomes available. In view of this, this report by WWF
International contributes to the further development of the IMO BWS Guidance with the aim
of triggering more BWM Convention ratifications in the near future, hopefully in 2013.
The BWS guidance presented in this report is based on results from earlier ballast water
sampling studies on ships voyages, also considering vessels ballasting patterns, and feasibility
of methods for PSC application. BWS methods for in-line (at discharge) sampling suggested
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in this BWS Guidance are based on the BWS methods developed and used for G8 type
approval testing of different BWMS around the world. In addition this report provides
background information for indicative versus detailed sampling. The sampling
recommendations given here may result in a workable, equitable and pragmatic solution.
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2 OBJECTIVES
The main objective of this study was to contribute to the ongoing discussions at IMO
regarding the procedures and methods to take samples resulting in representative numbers of
organisms in ballast water discharged. For representative sampling all organism groups
mentioned in the D-2 standard need to be addressed.
The D-2 standard states that ships meeting the requirements of the BWM Convention shall
discharge:
• less than 10 viable organisms per cubic meter greater than or equal to 50 micrometers in
minimum dimension, and
• less than 10 viable organisms per millilitre less than 50 micrometers in minimum
dimension and greater than or equal to 10 micrometers in minimum dimension, and
• less than the following concentrations of indicator microbes, as a human health standard:
• Toxigenic Vibrio cholerae (serotypes O1 and O139) with less than 1 Colony Forming
Unit (cfu) per 100 millilitres or less than 1 cfu per 1 gramme (wet weight) of
zooplankton samples,
• Escherichia coli less than 250 cfu per 100 millilitres, and
• Intestinal Enterococci less than 100 cfu per 100 millilitres.
Due to the lack of knowledge regarding representative ballast water sampling methods and
approaches, uncertainties exist how to assess compliance with the D-2 standard. The need for
a method resulting in a representative sample, and the difficulties in obtaining such a sample,
cannot be overstressed.
The main aim of this study is to provide additional BWS Guidance for sampling according to
the G2 Guidelines. This document covers sampling processes:
- “detailed sampling” to obtain a representative sample of the whole discharge (testing
for compliance),
- “indicative sampling”,
- “in-tank sampling” recommendations, and
- practical issues, such as possible sample concentration, sample transport etc.
Sample analyses processes and methods were not subject of this report and were therefore
excluded.
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3 MATERIALS AND METHODS
The BWS methods used in the research projects considered here were applied during practical
studies in which ballast water was sampled on vessels (e.g. Gollasch et al. 2002). During these
studies (see references for those studies the author was involved in) more than 1200 ballast
water samples were taken on more than 550 vessels of various types ranging from small cargo
vessels with deadweight of <1,000 tons to Very Large Crude Carriers (VLCCs) of >300,000
tons. The prime goal of these studies was to document the numbers and taxonomic variety of
organisms arriving in ports.
The sampling methods developed during these studies were consulted with an emphasis of
how the ballast water can be accessed. During these studies the ballast water samples were
taken using a variety of nets, hoses and pumps operated via tank openings (manholes), pumps
operated via sounding pipes, air vents (in-tank sampling), and by sampling water at the
sampling points which were installed for testing of BWMS (in-line sampling).
Very importantly, findings and experiences from three studies on representative BWS for
compliance monitoring, conducted for the Federal Maritime and Hydrographic Agency,
Hamburg, Germany in 2009 and 2012, and for the European Maritime Safety Agency, Lisbon,
Portugal in 2010, were in the focus of this study (Gollasch & David 2009, 2010, 2013). This
was complimented by the sampling experience gained on more than 60 shipboard tests for
type approval of 15 different BWMS which were conducted over the past 8 years.
For this contribution it was of prime importance to consider the appropriate sampling
approach for compliance control according to the BWM Convention, rather than to compare
different methods and sampling equipment, which was already published elsewhere (e.g.,
Gollasch et al. 2003).
The ballast water working group of IMO developed a sampling guideline, i.e., Guidelines for
Ballast Water Sampling (G2 Guidelines), adopted in 2008 (IMO, 2008a), which was also
considered when preparing this report. Furthermore, all developments at IMO regarding BWS
issues after the adoption of G2 were reviewed (IMO, 2008b-c, 2009a-g, 2010a-o, 2011a-h,
2012a-o, 2013) and taken into account. Special focus was given to:
• the aide-memoire on the analysis of samples developed at BLG13 (March 2009). BLG
agreed that guidance on sampling procedures could not yet be developed because of
the lack of relevant study results (BLG13/WP.5, Annex 1). This aide-memoire is
entitled “Initiate Development of an IMO Circular to provide Ballast Water Sampling
Analysis Protocols and to give Advice on Uniform Application of the Protocols” and
is limited to types of sample analysis, including recommendations for indicative and
detailed sample processing.
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• the preparational documents of the BWS circular (e.g., BLG15/5/1, BLG15/5/4-6).
This IMO Circular was aimed to give ballast water sampling and analysis guidance,
including sample representativeness and sample analysis protocols.
• the report of the Ballast Water and Biofouling Working Group from the last BLG
meeting (BLG17, February 2013), which contains the draft BWS circular (i.e.,
Guidance to Ballast Water Sampling and Analysis for Trial Use in Accordance with
the BWM Convention and Guidelines G2). The draft BWS circular was prepared for
consideration at MEPC65 (May 2013) and includes a collated list of sample analysis
protocols, methodologies and approaches for the D-1 and D-2 standards compliance
tests and recommendations for a trial period during which experience gained during
sampling will be used to update the BWS circular. However, it is stated in the
document that methods for representative sampling are still developing
(BLG17/WP.4) so that no details are included regarding the number of samples to take
or on their volume.
Additional sampling insights which contributed to the finding of this report were gained from
the author´s involvement in different national and international research studies, expert,
scientific and/or governmental working groups or organisations (i.e., ICES/IOC/IMO
Working Group on Ballast and Other Ship Vectors, Ballast Water Management Sub
Commission for the Adriatic Sea (BWMS), IMO/MEPC/Ballast Water Working Group,
Global Ballast Water Management Programme of GEF-UNDP-IMO, the European Maritime
Safety Agency and the relevant national authorities) where different aspects (i.e., biological,
technical, logistical) of ballast water sampling were addressed.
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4 BALLAST WATER SAMPLING GUIDANCE
4.1 COMPLIANCE CONTROL SAMPLING
After entry into force of the BWM Convention IMO Member States will be required to check
vessels for compliance with the standards of the BWM Convention, and this will be
undertaken by sampling ballast water on vessels. In accordance with Article 9.1, ships to
which the Convention applies may be subject to inspections for the purpose of revealing
violations of the provisions of the Convention. These inspections shall:
• Verify that the ship is carrying a valid Ballast Water Management Certificate;
• Verify that a Ballast Water Management Plan specific to the ship and approved by the
Flag State is onboard;
• Undertake an inspection of the Ballast Water Record Book.
As a part of Port State Control efforts and to demonstrate compliance with the BWM
Convention standards, port authorities may consider sampling ballast water for subsequent
analyses. The sampling guidance provided by IMO as in the Guidelines for Ballast Water
Sampling (G2) is based mainly on general information. Here the focus was laid on the
selection of appropriate sampling methodologies to assess compliance with the IMO ballast
water standards. Two standards are of particular relevance, i.e., the ballast water exchange
(Regulation D-1) and performance (Regulation D-2) standard.
The BWM Convention in the Regulation D-1, the Ballast Water Exchange Standard, states:
1 Ships performing Ballast Water exchange in accordance with this regulation shall do so
with an efficiency of at least 95 percent volumetric exchange of Ballast Water.
2 For ships exchanging Ballast Water by the pumping-through method, pumping through
three times the volume of each Ballast Water tank shall be considered to meet the standard
described in paragraph 1. Pumping through less than three times the volume may be accepted
provided the ship can demonstrate that at least 95 percent volumetric exchange is met.
Further requirements regarding ballast water exchange are given in Regulation B-4.
1 A ship conducting Ballast Water exchange to meet the standard in regulation D-1 shall:
.1 whenever possible, conduct such Ballast Water exchange at least 200 nautical miles from
the nearest land and in water at least 200 metres in depth, taking into account the Guidelines
developed by the Organization;
.2 in cases where the ship is unable to conduct Ballast Water exchange in accordance with
paragraph 1.1, such Ballast Water exchange shall be conducted taking into account the
Guidelines described in paragraph 1.1 and as far from the nearest land as possible, and in all
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cases at least 50 nautical miles from the nearest land and in water at least 200 metres in
depth.
2 In sea areas where the distance from the nearest land or the depth does not meet the
parameters described in paragraph 1.1 or 1.2, the port State may designate areas, in
consultation with adjacent or other States, as appropriate, where a ship may conduct Ballast
Water exchange, taking into account the Guidelines described in paragraph 1.1.
Unlike sampling for approval of ballast water treatment systems, which need to meet the
above mentioned D-2 standard, sampling for compliance control is required to identify any
possible non-compliance with the D-2 standard.
If the sampling must demonstrate compliance with the D-2 standard, then documenting the
number of organisms greater than or equal to 50 micrometres in minimum dimension is
especially challenging since less than 10 viable organisms per m³of water are acceptable.
Various difficulties can be identified such as more than 1,000 l of water may need to be
collected to proof compliance, and several replicates need to be sampled to meet general
scientific standards and accuracy. The accuracy of the sampling technique must be
determined; inefficient sampling techniques may result in false negatives as a result of
missing organisms.
Different vessel specifics, such as types, sizes and cargo profiles result in very different
ballast water discharge profiles and times. The ballast water discharge of a vessel may be
conducted “at once” or “in sequences” which may last from approximately one hour, e.g.,
discharge of two tanks in parallel on a container vessel, up to several days. The tank discharge
duration is depending on the length of the cargo operation, e.g., tankers, bulk carriers and
sometimes also general cargo vessels, load cargo over several days duration. Hence, the
ballast water operation is frequently conducted in sequences over the time of the cargo
operation. This factor is important to be taken into account as it is difficult to assume that the
PSC officer and/or sampling team will stay on-board a vessel for several days.
4.2 SAMPLING METHODS ACCORDING TO THE ACCESS POINT
Sampling points may be divided into in-tank and in-line (at discharge) sampling points. In-
tank sampling points enable ballast water access directly from a tank either via ballast tank
manholes, sounding or air pipes. In-line sampling points include the ship´s pipe work after the
ships ballast water pumps.
For compliance monitoring with the D-1 standard in-tank or in-line samples may be taken to
either proof the presence of coastal biota or check the water salinity. This may be done by
using all possible access points including sounding pipe, manhole and the vessels main ballast
water line. However, the use of the latter is not recommended because a discharge to sea may
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occur during sampling and in case of non-compliance the non-complaint water would enter
the recipient environment during the sampling event. As the D-1 standard is not a numerical
standard no quantitative biological sampling is needed, but a qualitative approach to
document the possible occurrence of coastal biota.
In contrast, the D-2 standard, being a numerical and biological discharge standard the samples
should be taken from the ballast water discharge line (but see below). Here, a quantitative
biological approach is needed as the numerical standard applied to living organisms above 10
micrometres in minimum dimension no matter what type they are. For the indicator microbes
as stated in the D-2 standard a qualitative and quantitative approach is needed so that the
concentration of colony forming units of certain indicator microbes becomes known.
Although in-line sampling seems most appropriate to assess the D-2 standard compliance as
discharge standard, in case the ballast water originates from a high risk area, i.e., an area with
a known occurrence of target species, samples may be taken from the ballast tank prior
discharge. This enables non-compliance actions before the water was discharged into the
recipient environment. The in-tank approach is also advisable to proof compliance with the D-
2 standard for tanks which have direct discharge to sea, e.g., top-side tanks on some bulk
carriers.
4.3 TANK SELECTION – WHICH TANK TO SAMPLE (FIRST)?
Vessel may have ballast water to be discharged from different sources and also with different
uptake dates (holding time on board). If possible, ballast water from all different sources
might need to be tested. If this is impossible or in terms of interest to have a result as soon as
possible, possibly prior any discharge, tank(s) to be sampled first should be selected based on
a risk assessment. This risk assessment will focus on the indication which ballast water may
contain potentially harmful species for the recipient port.
Such a risk assessment may consider the following elements, but may not be limited to:
• the environmental compatibility of the ballast water source area with the ballast water
recipient area;
• the presence of harmful aquatic organisms and pathogens in the ballast water origin
area;
• if appropriate, the presence of target species in the ballast water origin area; and
• the in-tank holding time.
The tank(s) with higher environmental compatibility of the discharge area with the ballast
water origin, tanks with ballast water origin area where Harmful Aquatic Organisms and
Pathogens (HAOP) or target species are present, and tanks with shorter in-tank holding time
should be given priority for ballast water sampling as these would likely pose the highest risk
to introduce HAOP.
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In the case of in-tank sampling, 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 (David and Perkovič, 2004).
For in-line sampling the tank(s) which is(are) currently discharged when PSC comes on board
may be sampled first, not to make PSC wait many hours until the »targeted tank« is ready to
be discharged.
In general, tanks being discharged or those to be discharged first may be given priority as this
would give the opportunity to PSC officers to decide for appropriate management measures in
case non-compliance, or an indication of possible non-compliance, is identified.
4.4 SAMPLING FOR COMPLIANCE WITH THE D-1 STANDARD
To prove that the ballast water was exchanged, salinity measurements of the ballast water may
verify if the water was exchanged according to the BWM Convention requirements. It is
suggested that in cases where the salinity is low, e.g., below 30 psu, it can be assumed that the
ballast water originates from coastal areas with freshwater influence so that it was not
exchanged with ocean water, i.e., outside 50 or 200 nautical miles from nearest land and at
water depths higher than 200 metres, because this water would have a higher salinity.
Consequently, this D-1 standard compliance control option would only deliver results when
the inspected vessel has loaded ballast in a lower salinity or freshwater port.
The D-1 standard requires that at least 95 percent of the water needs to be exchanged
therefore up to 5 % of water may be unexchanged. When a vessel has taken up ballast water
in a freshwater port (100% tank volume) and 95% are exchanged in mid ocean, 5% freshwater
may have remained in the tank during the exchange process and will dilute the exchanged
ocean water salinity. As a consequence a false non-compliant indication may occur because
the remaining freshwater from the previous tank filling would be ignored.
The lowest seasonal averaged ocean salinity is ca. 30 psu (see Figure 1). Consequently,
should during a D-1 standard compliance control test the measured salinity of a mid-ocean
exchanged ballast tank be less than 30 psu, it is likely that the ballast water in this tank
originates from coastal areas with freshwater influence. Ballast water exchange resulting in
salinities less than 30 psu likely occurred less than 50 nautical miles from the nearest shore
with freshwater influence from nearby rivers or estuaries, as otherwise the salinity should be
higher. Such a low salinity measurement indicates that the ballast water was exchanged closer
to land than required.
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Figure 1 - Annual world ocean´s surface salinity (Source:
http://www.ccpo.odu.edu/
~arnoldo/ocean405/s00anw01.gif).
Sampling for the presence of coastal biota as D-1 standard compliance control is also of a
limited value because very few organisms are restricted in their occurrence to coastal waters
alone. Candidate species to consider for such an approach include harpacticoid copepods and
barnacles. Most barnacles are fouling species and the majority of them are found along the
tidal zone of hard-bottom shorelines. Therefore their presence in a ballast water sample
indicates coastal origin of the water. Barnacles are also frequently found in the biofouling of
vessels. In the unlikely event that two vessels follow each other closely the barnacles in the
biofouling of the first vessel may release their larvae into the water which may be pumped
into a ballast tank of the second vessel during the ballast water exchange operation. In this
case the exchanged ballast water, even when the exchange was carried out in mid-ocean, may
contain coastal organisms from the biofouling of the first vessel. Barnacles also occur inside
ballast tanks fouling on the tank walls. In theory they may also reproduce inside the tank and
release their larvae into the ballast water. The presence of these larvae from in-tank
reproduction may then be wrongly identified as originating from a ballast water exchange in
coastal areas. This scenario is of low probability, but it cannot be excluded completely. The
other organism group, harpacticoid copepods, are benthic species so that their presence in
ballast water indicates coastal origin of the water. However, these species were also
frequently found in the ballast tank sediment. In addition, a study showed that the numbers of
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the harpacticoid copepods in a ballast tank increased during a voyage which indicated that
reproduction may have occurred (Gollasch et al. 2000).
The presence of human faecal bacteria, such as Escherichia coli, Enterococci or Vibrio
cholerae may also be used to document compliance with the D-1 standard. Their presence
indicates improper waste water management in/close to urban areas. As most of these
indicator microbes are not surviving for longer times as free living in marine waters their
occurrence in a ballast tank sample indicates near shore ballast water exchange, i.e., the depth
and distance requirement of the D-1 standard was not meet when undertaking the water
exchange. As these indicator microbes are unlikely to survive a (longer) vessel voyage in a
ballast tank without their human (or other) “hosts” this method seems less reliable for longer
voyages.
As shown above, an analysis of the biota in a tank for compliance control with the D-1
standard only delivers results with a limited level of certainty. Therefore, in cases non-
compliance is indicated by these methods, it is assumed that these data are not robust enough
to enable a non-compliance action.
Another D-1 standard compliance control option may be to document the human influence on
the sea. It seems to be logical that this is greater in near shore regions compared to the high
seas. Candidate methods include to document tracers of human activity, such as high Nitrogen
or Phosphorous levels which may result from river run-offs in areas with human settlements.
However, this method may deliver regionally very different results. In several oceanic regions,
especially in island states or in costal environments with lower human populations, the
Nitrogen or Phosphorous levels may be very low even in areas close to shore. However, it
was concluded that several trace elements may be used to identify the oceanic origin of ballast
water which can be used to evaluate whether or not ballast water was exchanged at sea
(Murphy et al. 2004, Hunt et al. 2007). Consequently a tracer detection tool was developed for
compliance control (Murphy et al. 2008).
Murphy et al. (2006) further showed that fluorescence may be used to verify ballast water
exchange for most samples of high salinity ballast water. However, water contamination with,
e.g., fuel oil, may impair the fluorometric measurements.
Optical instruments may also be used to analyse the characteristics of chromophoric dissolved
organic matter (CDOM) in water (Murphy et al. 2008). CDOM is a result of decayed
(terrestrial) plants and it was assumed that higher CDOM concentration indicate near shore
ballast water exchanges.
Because of all listed limitation, biological sampling is not recommended for D-1 standard
compliance control. The other candidate methods also seem to generate results with lower
confidence levels so that the measurement of ballast water salinity seems to be the most
reliable option.
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4.4.1 IN-TANK D-1 STANDARD SAMPLING
4.4.1.1 Selection of ballast water sampling methods for in-tank D-1 standard sampling
As suggested here, the in-tank D-1 standard compliance control sampling is limited to non-
biological analysis of the water. It is explained above that salinity seems to be a good
parameter for this analysis. Already a very small volume of water enables this measurement.
The water for salinity measurement may be collected by a water column sampler or pump.
Alternatively the sensor of a salinity meter may be lowered into the tank or sounding pipe to
enable a direct measurement.
However, measuring the salinity from ballast tanks is not as trivial as thought. Salinity
measurements via sounding pipes with separate readings in different heights documented that
the salinity was not homogenous over the length of the sounding pipe. The deeper the salinity
meter was lowered in the sounding pipe the higher the salinity was. This is a clear indication
that more than one sample may need to be taken to cover potentially varying salinities of
ballast water in the same tank. This may either be done by lowering the sensor of a salinity
meter to different depths in a ballast tank or by taking multiple salinity meter readings from
different depths in a sounding pipe.
The suggested sampling methods and equipment are outlined in Table 1. A detailed
description of the sampling equipment and sampling arrangements is in chapter 4.8.
Table 1 - Possible sampling access points, equipment and other details recommended for
compliance control sampling with the D-1 standard.
Sampling point
Equipment
Water volume
Number of samples
Sounding pipe,
manhole or air vent
Water column
sampler or pump
> 50 ml
1 integrated sample
from possibly whole
water column
Sounding pipe,
manhole or air vent
Point-source
sampler or pump
> 50 ml
1 integrated sample
from 3 different
depths
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4.4.1.2 Description of sampling methods for in-tank D-1 standard sampling
In-tank D-1 standard sampling requires relatively small quantities of water to be sampled,
hence the easiest way may be to use simple water samplers, e.g., water column sampler or
pump. Alternatively a salinity meter may be lowered to the desired water depth for measuring
the salinity.
4.4.1.2.1 Water column sampler
To obtain the integrated sample from the whole water column, the water column sampler is to
be lowered possibly to the bottom of the tank or to the deepest point accessible. When
lowering the water column sampler, water will start entering the sampler from the opening at
the top of the sampler. The water will be proportionally sampled from the entire water column
provided the sampler is lowered through the water column with a constant speed, and the time
used to lower the sampler from the surface water to the deepest point accessible of the tank is
the same as the time the sampler is filled with water. As a relatively low water volume is to be
sampled, i.e., >50ml, the water column sampler may need to be lowered down only once.
4.4.1.2.2 Point-source sampler
To obtain three samples from different depths, the point-source sampler is to be lowered three
times to different depths, i.e., the surface, somewhere in the middle of the water column, and
possibly close to the tank bottom or to the deepest point accessible. Each time the valve is
opened by pulling the rope connected to the valve at the bottom of the sampler to make the
water enter the sampler. The valve is closed again when the rope connected to the valve is
relaxed, and then the sampler is pulled up. The three samples from different depths are
integrated, i.e., mixed together, and one salinity value is measured. As relatively low water
volume is to be sampled, i.e., >50ml, point-source sampler may need to be lowered only once
per desired depth, all together three times.
4.4.1.2.3 Pump
A pump can be used to obtain an integrated sample from three different depths or from the
whole water column. The pump, or the suction side of the hose connected to the pump, is to
be lowered down to three desired depths, i.e., the surface, somewhere in the middle of the
water column, and possibly close to the tank bottom or to the deepest point accessible, and
from each depth water is to be pumped up. Alternatively, when lowering the pump or the
suction side of the hose connected to the pump, water is to be pumped up constantly from the
surface water to the deepest point accessible. The limiting factor to be considered is the
pumping head of the pump. As relatively low water volume is to be sampled, i.e., >50ml, only
short pumping time is needed from each of the three desired depths or constantly during
lowering the pump to the deepest point accessible in the tank.
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4.4.2 IN- LINE D-1 STANDARD SAMPLING
4.4.2.1 Selection of ballast water sampling methods for in-line D-1 standard sampling
In-line ballast water sampling for the D-1 standard would be probably of very low probability
as vessels that do not have BWMS installed also do not have in-line sampling points.
Nevertheless, in case there would be a sampling point somewhere in the line, maybe also a tap
at the ballast pump, this could be the chosen approach, especially in case the discharge of
ballast water was already started and is ongoing.
However, having in mind that D-1 standard sampling may also be used to have an early
indication of potentially non compliant ballast water, and consequently to apply appropriate
management measures, one key problem remains, and this is that compliance or non-
compliance can in this scenario only be proven while the ballast water is being pumped
overboard. Should, e.g., a risk assessment result in the identification of high risk ballast water
to be discharged the in-line sampling during discharge should be avoided, but in-tank
sampling should be undertaken to assess compliance.
The suggested sampling method and equipment is outlined in Table 2. A detailed description
of the sampling equipment and sampling arrangements is in chapter 4.8.
Table 2 - In-line sampling equipment and other details recommended for compliance control
sampling with the D-1 standard.
Sampling point
Equipment
Water volume
Number of samples
In-line
Sampling bottle
> 50 ml
1 sample as soon as
possible during the
discharge
Although it is known that some salinity stratification may occur in the tank, and hence
multiple measurements, e.g., one at the beginning, one in the middle, and one in the end of
discharge, could identify such differences, such measurement could be very impractical
especially when ballast water is discharged over longer time. Secondly, this would also not be
appropriate in the interest of having the result as soon as possible before all possibly non
compliant ballast water is discharged.
4.4.2.2 Description of sampling methods for in-line D-1 standard sampling
A small sample bottle is sufficient. Should a conductivity meter be used, it is recommended to
use bottles with a wider opening that the conductivity sensor can be inserted to the sample
right away.
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4.5 SAMPLING FOR COMPLIANCE WITH THE D-2 STANDARD
Compliance control with the D-2.1 standard is solely quantitative, thereby ignoring the type of
organisms with the exception of selected indicator microbes (D-2.2). It is the numbers of
living organisms per size class to document (non-)compliance. As the BWM Convention in
Regulation D-2 reads "…Ships conducting Ballast Water Management in accordance with
this regulation shall discharge less than 10 viable organisms per m³ …" the D-2 standard is
understood as discharge standard, what indicates that the most suitable sampling point to
proof D-2 standard compliance is the discharge line of the vessels ballast water system as also
recommended in the Guidelines G2, i.e. samples should be taken from the discharge line, as
near to the point of discharge as practicable, during ballast water discharge whenever
possible.
However, in-tank sampling should also be considered as a valid option because some vessels,
e.g., some bulk carriers and tankers, may have upper side wing tanks that are emptied via
direct overboard discharge valves rather than by using ballast pumps (Figure 2). In such cases,
the Guidelines G2 indicate that in-tank sampling may be an appropriate approach.
Figure 2 - Ballast water discharge above pier level from the upper wing tanks of a bulk carrier
(Jure Barovic, with courtesy of the Port of Koper, services for protection of the sea).
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Compliance tests for the D-2 standard can only be made by sampling for the biological
content of the ballast water.
4.6 INDICATIVE SAMPLING
The indicative sample analysis is described in the Ballast Water Sampling Guideline G2. The
paragraph 6.3 reads: 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.
For ballast water sample analyses, certainly, as a very first step, a sampling event needs to be
conducted. However, the G2 does not address explicitly how an indicative sampling event
would need to be undertaken. Implicitly, an indicative analyses could be performed on a
sample, or part of it, taken during the detailed D-2 standard compliance control sampling
process, or just on any stand-alone sample.
One important point is that an indicative sampling event may be targeted towards only one
group of organisms, i.e., organisms less than 50 micrometres in minimum dimension and
greater than or equal to 10 micrometres in minimum dimension or organisms greater than or
equal to 50 micrometres in minimum dimension or indicator microbes included in the D-2
standard. The results from each of these organism groups alone may already give an
indication that a BWTS is not performing properly. From the author’s experience of on-board
sampling for type approval of BWMS, it is likely that, indicator microbes and organisms less
than 50 micrometres in minimum dimension and greater than or equal to 10 micrometres in
minimum dimension meet the D-2 standard, however organisms greater than or equal to 50
micrometres in minimum dimension may be found in too high concentrations and exceed the
acceptable organism numbers in the D-2 standard.
Different groups of organisms in general require different sampling approaches. In general
organisms greater than or equal to 50 micrometres in minimum dimension require larger
water quantities to be sampled to collect them compared to when focussing on organisms less
than 50 micrometres in minimum dimension and greater than or equal to 10 micrometres in
minimum dimension. This is because there are relatively lower concentrations of organisms
greater than or equal to 50 micrometres in minimum dimension in the water as for organisms
less than 50 micrometres in minimum dimension and greater than or equal to 10 micrometres
in minimum dimension. Consequently, indicative sampling methods may be very different for
each organism group, and may differ in, e.g., sampling duration, timing, volume, and at which
sampling point it should be taken.
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Without a known history of the performance of a certain BWMS it is very difficult to predict
in advance which group of organisms to sample for to identify possible non-compliance with
the D-2 standard. This would require a risk assessment to be conducted in advance. Therefore,
it would be most effective to use a sampling method enabling analyses on all organism groups.
This would also enable a step-by-step process, where one analysis method for one organism
group may be applied first, and in cases this shows some indication or even does not give an
indication of non-compliance, the second organism group may be tested for with another
sample analysis method.
Another issue to consider is the consequences which may arise from an indicative analysis.
Does an indicative result trigger the need for further tests, i.e. a detailed D-2 standard
compliance test? or Should a vessel be banned from discharging ballast water solely based on
the indicative result? Based upon paragraph 6.3 of the G2 guideline, it is understood that an
indicative analysis was included in G2 to give a Party an opportunity to identify potential non-
compliant ballast water in an early stage, i.e., as the detailed compliance test is expected to
show results only after all ballast water was already discharged, to avoid any additional
impact from a possibly non-compliant discharge from a ship.
4.6.1 IN-TANK INDICATIVE SAMPLING
It is expected that sampling will be conducted on a number of various types of ships in a port.
After having selected a ship according to the sampling program, i.e., targeting of vessels
based on PSC, the ballast tanks are to be selected for sampling. Sampling access plays a
crucial role and actually determines if the ballast water is available for sampling. Therefore,
sampling equipment, which allows access to various sampling points, is in most cases crucial
to obtain a sample.
Another aspect is the need for in-tank sampling. On occasion it may be appropriate not to take
a sample from the ballast water discharge line as G2 recommends during the discharge
overboard, if it is known that a vessel carries ballast water from areas with outbreaks,
infestations, or populations of HAOP, e.g., toxic algal blooms, the sampling during the
discharge should be avoided. Risk assessment should be used to identify high risk ballast
water.
Such ballast water discharges overboard, pose a risk to the environment, human health,
property or resources. Instead it is recommended that in such cases it is preferred to take an
indicative ballast water sample directly from the ballast water tank prior any discharge
of ballast water. Although such sampling events may not be representative of the whole
discharge, an indicative compliance control test is enabled without taking the risk to discharge
the ballast water into the environment.
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4.6.1.1 Selection of ballast water sampling methods for indicative in-tank sampling
It was concluded that sampling for zooplankton via the sounding pipes does not result in a
representative sample of species in the tank as comparisons of sounding pipe and manholes
samples from the same tank found that net samples deliver more diverse biological results.
Sounding pipe samples contained 0-60% of the organisms of a net sample indicating the need
to sample ballast tanks via opened manholes. Further, pumps used via open manholes
delivered more diverse samples than net samples, therefore pumps may also be considered
when sampling via manholes. Future indicative compliance control in-tank studies should
take into account that sampling via sounding pipes is inferior when selecting appropriate
sampling techniques. However, frequently manholes cannot be opened due to, e.g., overlaying
cargo or on-going cargo operations in the area where the manhole is located, and in these
instances sounding pipe sampling might be the only solution to sampling.
The suggested sampling methods and equipment are outlined in Table 3. A detailed
description of the sampling equipment and sampling arrangements is given in chapter 4.8.
Table 3 - Possible sampling access points, equipment and other details recommended for
indicative compliance control in-tank sampling with the D-2 standard.
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
whole water
column or from 3
different depths
< 50 and > 10
micrometres
Manhole,
sounding pipe,
manhole or air
vent
Pump or water
column sampler
5 – 6
1 integrated
sample from
possibly the
whole water
column or from 3
different depths
Manhole,
sounding pipe,
manhole or air
Pump or point-
source sampler
5 - 6
1 integrated
sample from 3
different depths
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vent
Indicator
microbes
Manhole,
sounding pipe,
manhole or air
vent
Pump or water
column sampler
1
1 integrated
sample from
possibly the
whole water
column
Manhole,
sounding pipe,
manhole or air
vent
Pump or point-
source sampler
1
1 integrated
sample from 3
different depths
4.6.1.2 Description of sampling methods for in-tank indicative sampling
For organisms greater than or equal to 50 micrometres in minimum dimension a plankton net
is suggested to be used because of the ease to filter the suggested 300-500 litres. As a second
choice a pump can be used to pump up and filter 100 litres. However, if a tank would be
accessible only through the sounding or air pipe, a pump would probably be the only
appropriate choice.
For the organisms less than 50 micrometres in minimum dimension and greater than or equal
to 10 micrometres in minimum dimension, and for indicator microbes only small quantities
need to be sampled, hence a pump or different water samplers may be used. In case the pump
is already used for sampling for organisms greater than or equal to 50 micrometres in
minimum dimension, 100 litres may be filtered, and at the same time additional 5 – 6 litres of
a continuous drip sample may be collected in a bucket for organisms less than 50 micrometres
in minimum dimension and greater than or equal to 10 micrometres in minimum dimension
and for indicator microbes. The indicator microbes sample of 1 litre may be subsampled from
the bucket.
4.6.1.2.1 Plankton net for in-tank sampling?
Lower the net to the maximum possible level in the tank, wait a minute and retrieve the net
with an approximate speed of 0.5 m per second. Empty cod-end into sample bottle and repeat
the sampling procedure to meet the desired water volume.
4.6.1.2.2 Pumps
Pump can be used to obtain an integrated sample from 3 different depths or from the whole
water column. Pump is to be lowered down to 3 desired depths, i.e., the surface, somewhere
in the middle of the water column, and possibly close to the tank bottom or to the deepest
point accessible, and from each depth water is to be pumped out. Alternatively, when
lowering the pump, water is to be pumped out constantly from the surface water to the deepest
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point accessible. The limiting factor to be considered is pumping head of the pump. As
relatively low water volume is to be sampled, i.e., >50ml, only short time pumping is needed
from each of the 3 desired depths or constantly during lowering the pump to the deepest point
accessible in the tank.
4.6.1.2.3 Water-Column Sampler
To obtain the integrated sample from whole water column, the water column sampler is to be
lowered possibly to the bottom of the tank or to the deepest point accessible. When lowering
the water column sampler, water will start entering the sampler from the opening at the top of
the sampler. The water will be proportionally sampled from the entire water column provided
the sampler is lowered through the water column with a constant speed, and the time used to
lower the sampler from the surface water to the deepest point accessible of the tank is the
same as the time the sampler is filled with water. As relatively low water volume is to be
sampled, i.e., >50ml, the water column sampler may need to be lowered down only once.
4.6.1.2.4 Point-Source Sampler
To obtain 3 samples from different depths, the point-source sampler is to be lowered 3 times
to desired depths, i.e., the surface, somewhere in the middle of the column, and possibly close
to the tank bottom or to the deepest point accessible. Each time the valve is opened by pulling
the rope connected to the valve at the bottom of the sampler to make the water enter the
sampler. The valve is closed again when the rope connected to the valve is relaxed, and then
the sampler is pulled up. The 3 samples are integrated (mixed together) and one salinity value
is measured. As relatively low water volume is to be sampled, i.e., >50ml, point-source
sampler may need to be lowered only once per desired depth, all together 3 times.
4.6.2 IN- LINE INDICATIVE SAMPLING
4.6.2.1 Selection of ballast water sampling methods for indicative in-line sampling
Considering all the above and especially that the in-line sampling test may develop also in
detailed D-2 standard compliance test, it is recommended that for an in-line indicative
ballast water sampling event, one sequential sample is taken using the same sampling
methodology as for a detailed D-2 standard compliance test (see 4.7.2 and 4.7.3).
To take one sequential sample a relatively short sampling time is sufficient and the sample
analysis may be conducted with a variety of different methods. The results obtained in this
way can also represent very solid grounds for different actions which PSC may have available
in case of non-compliance is indicated. These include:
• the requirement that more tests are needed and to proceed to a detailed compliance D-
2 standard test,
• sending the vessel to a designated ballast water discharge area,
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• to require ballast water discharge in a port reception facility, or even
• to ban the vessel from further ballast water discharge.
The decision to take would be depending on the sampling result obtained. For instance, if the
organism concentration identified is just above the D-2 standard, this would indicate possibly
that further tests are required. In contrast, should the organism concentration be much higher
than the D-2 standard, i.e., gross exceedence, a vessel may be banned from continuing the
ballast water discharge, may be sent to a designated ballast water discharge area, or required
to discharge ballast water into a port reception facility.
The suggested methods and equipment are outlined in Table 4. A detailed description of the
sampling equipment and sampling arrangements is given in chapter 4.8.
Table 4 - Possible sampling access points, equipment and other details recommended for
indicative in-line compliance control sampling with the D-2 standard.
Organism
group
Sampling point
Equipment
Water volume
[litre]
Number of
samples
> 50
micrometres
In-line
Plankton net
300 – 500 in
each sequence
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
Bucket
5 – 6
1 continuous drip
sequential
sample, may be
simultaneously
collected during
sampling of
organism group
> 50
micrometres
Indicator
microbes
In-line
Bucket
1
1 continuous drip
sequential
samples, may be
sub-sampled
from the bucket
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4.6.2.2 Description of sampling methods for indicative in-line sampling
It is recommended that for an in-line indicative ballast water sampling event, one sequential
sample is taken using the same sampling methodology as for a detailed D-2 standard
compliance test (see 4.7.2 and 4.7.3).
4.7 DETAILED SAMPLING FOR COMPLIANCE WITH THE D-2
STANDARD
4.7.1 IN-TANK D-2 SAMPLING
As D-2 is a discharge standard, in-tank sampling for this purpose is of limited value. However,
should an in-tank sample reveal very high organism numbers, non-compliance may be
assumed also when the water is discharged. To illustrate this, should a sample from the tank
contain 1000 organisms and the tank capacity is 100 tons, the organism concentration would
exceed the D-2 standard when the water is discharged.
4.7.2 IN- LINE D-2 STANDARD SAMPLING
4.7.2.1 Selection of sampling equipment and methods for in-line D-2 standard sampling
Previous studies have shown that different approaches, i.e., short/long sampling times, in the
sampling process result in different living organism concentrations (Gollasch & David 2009,
2010, 2013). Therefore the selection of an inappropriate sampling approach will influence the
compliance control result. As a consequence the living organism concentrations in the ballast
water discharge may therefore be underestimated, so that an inefficient BWMS could be
recognised as compliant. In contrast, living organism concentrations may be also
overestimated, and a BWMS complying with the D-2 standard may fail in compliance tests.
The sequential trials showed different organism numbers in each sequence of one test
indicating the patchy organism distribution inside the ballast tank. This was observed during
all sampling events and for both organism groups studied. Hence, sampling during ballast
water discharge is biased by tank patchiness of organisms.
In the group of organisms greater than or equal to 50 micrometres in minimum dimension the
previously undertaken studies showed that samples taken over the entire discharge time of a
tank contained much lower living organism concentrations compared to the living organism
count in the sequences so that sequential sampling may deliver more representative results.
Comparative studies have shown that sequential samples of approximately 10 minutes
duration are suitable for in-line D-2 standard compliance tests. In most tests the highest
zooplankton count occurred in the last sequence so that sampling at this time may “over
sample” the organism concentration. It was further observed that sequential samples taken in
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the very beginning and end during a ballast tank is emptied are unlikely to give representative
results of the living organism concentration because in these samples the organism count
showed high variations, which may result in under- or oversampling the organism
concentration. Even when avoiding these time windows the concentration of living organism
still seems to be patchy so that it is recommended to take at least two sequential samples in
this time window. The mean value of the living organism concentration in these two samples
may be taken to assume the real organism concentration.
In the group of organisms less than 50 micrometres in minimum dimension and greater than
or equal to 10 micrometres in minimum dimension cell counts of sequences in comparison to
the samples taken over the entire time of a tank discharge showed lower living organism
concentrations in sequential samples, which is in contrast to the zooplankton results. The
comparison of the phytoplankton cell concentration between the different sequences of all
tests showed that no clear trend can be identified during which time window a more
representative sample will be taken. Therefore it is recommended to take at least two
sequential samples during the discharge of a ballast water tank with avoiding taking a sample
during the very beginning and end of the discharge time of a tank or tanks. The mean
organism count in these two or more samples may be seen as the real living organism
concentration in the ballast water.
The suggested methods and equipment are outlined in Table 5. A detailed description of the
sampling equipment and sampling arrangements is given in chapter 4.8.
Table 5 - Possible sampling access points, equipment and other details recommended for
compliance control sampling with the D-2 standard.
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
Bucket
5 – 6 in each
sequence
2 (or more)
continuous drip
sequential
samples
collected at the
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same time as for
organism group
> 50
micrometres
Indicator
microbes
In-line
Bucket,
sampling bottle
1 in each
sequence
2 (or more)
continuous drip
sequential
samples sub-
sampled from the
bucket
4.7.3 RECOMMENDATIONS FOR A BALLAST WATER SAMPLING PROTOCOL THAT
IS REPRESENTATIVE OF THE WHOLE DISCHARGE
4.7.3.1 Samples representativeness
The results from previous ballast water sampling studies have shown that different approaches
in the sampling process have an influence on the results regarding organism concentrations
(Gollasch & David 2009, 2010, 2013). The organisms are potentially affected, so that the
selection of the “wrong” sampling approach may influence the compliance control sampling
result. As a consequence, the organism concentrations in the ballast water discharge may be
underestimated, and a not well performing BWTS could falsely become recognised as
compliant. In contrast organism concentrations may be overestimated so that and a BWTS
which complies with the D-2 standard may fail the compliance test.
A certain level of pragmatism is required during on-board ballast water compliance control
sampling as the work is not undertaken under controlled laboratory conditions. This is
especially relevant when sampling for organisms greater than or equal to 50 micrometres in
minimum dimension. All attempts should be made to avoid negative impacts of organism
survival during the sampling process. PSC are unlikely to have larger water collecting tanks,
e.g., >500 litres, available during the sampling event and will therefore probably need to work
with nets to concentrate the sample during the sampling procedure. The Ballast Water
Sampling Guidelines (G2) also includes these aspects: sampling should be undertaken in a
safe and practical manner; and samples should be concentrated to a manageable size.
It was previously observed that the sampling duration (i.e., length of the sampling process),
the timing (i.e., the point in time during the ballast water discharge the sampling is conducted),
the number of samples and the water quantity sampled are the main factors to influence
organism concentrations results (Gollasch & David 2009, 2010, 2013).
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4.7.3.1.1 Recommended sampling duration
It was shown that the organisms greater than or equal to 50 micrometres in minimum
dimension are negatively affected by longer sampling times. Considering that the results also
showed that a shorter sampling time results in a representative sample, the recommended
sampling time of a sequential sample is approximately 10 minutes. Longer sampling times
are likely to result in an underestimation of the viable organism concentration in the ballast
water discharged, especially for organisms greater than or equal to 50 micrometres in
minimum dimension.
4.7.3.1.2 Recommended sampling timing
Organism concentrations are likely to vary considerably if the sampling event is conducted at
the very beginning or at the very end of the ballast water discharge process (or tank) because
at these times the patchy organism distribution inside ballast water tanks was the greatest.
Therefore, it is not recommended to take a sample at the first 5 minutes or at the last 5
minutes of the ballast water discharge. An underestimation or an overestimation of organism
concentrations may have to be expected. It is therefore recommended that the sampling is
conducted random sequence of approximately 10 minutes duration anytime in the
middle of the ballast water discharge from a tank or tanks, starting not before 5 minutes
from the start of discharge and ending not after 5 minutes before the end of the
discharge.
4.7.3.1.3 Recommended number of samples
It was previously documented that the organism concentration of all organism groups in the
D-2 standard varies due to the patchy distribution inside the ballast water tanks. Due to this
variation a single 10 minute sequential sample may underestimate or overestimate the
organism concentration discharged. It was also observed that the average organism
concentrations of 2 random sequential samples provide very similar results to the average of 3
random samples. In consequence it is recommended that sampling is carried out by
undertaking at least 2 random samples, and that the samples are analysed immediately after
each sampling event has ended. For the final result the organism concentration results of
the sequences sampled are averaged.
4.7.3.1.4 Recommended sampled quantity
In earlier studies (Gollasch & David 2009, 2010, 2013) sequential sampling was conducted
over different time periods of the sequences, i.e., 5, 10 and 15 minutes, with the water flow
rate averages ranging from 30 to 50 litres per minute. To obtain most representative results it
is recommended that:
• for the organisms greater than or equal to 50 micrometres in minimum
dimension between 300 and 500 litres should be filtered and concentrated;
• for the organisms less than 50 micrometres in minimum dimension and greater
than or equal to 10 micrometres in minimum dimension a "continuous drip"
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sample with a total volume of not less than 5 litres should be taken, e.g., collection of
about 0.5 litres of sample water every minute during the entire sequential sampling
time duration; alternatively 0.5 litres of sample water may be collected every 30 to 45
litres of the ballast water sampled, depending on the flow rate. The resulting 5 litres of
collected sample water should be mixed and sub-sampled in two sets, one set of
samples alive and another preserved. As sub-sample volume approximately 60 to 100
ml are recommended;
• for indicator microbes samples, a sample of approximately 1 litre should be
collected as a sub-sample after mixing from the 5 litre continuous drip sample.
4.7.3.1.5 Other recommendations
It was further assumed that the sampling flow rates may have an additional influence on the
concentration results of viable organisms. In case lower flow rates are obtained by partially
closing valves of the sampling line this may result in sheer forces which likely will damage
organisms during the sampling process. A similar negative effect may be caused by very
strong flow rates, which is affecting mainly the organisms greater than or equal to 50
micrometres in minimum dimension. Hence, the flow rate or “valve” effect, may cause an
underestimation of viable organisms as organisms may have died during the sampling process.
To avoid this negative effect it is recommended that the valve at the sampling point is opened
as much as possible. However it should not exceed the flow rate of 50 litres/min so that the
water pressure in the sampling net is not too high during sample concentration as this may
negatively affect organism survival.
4.7.4 SAMPLING LOGISTICS FEASIBILITY
It is well known that vessels of different types, sizes and cargo profiles trigger very different
ballast water discharge patterns. The ballast water discharge may be carried out as a one time
event “at once” or over longer time durations “in sequences”. These may last from
approximately one hour, e.g., a fast discharge of two tanks in parallel on a container vessels.
Longer discharge durations may stretch up to several days depending on the length of the
cargo operation. This may be the case on tankers, bulk carriers and sometimes general cargo
vessels which load cargo during several days. Hence the ballast water operation is frequently
conducted in sequence over the time of cargo operation.
This factor is important to be taken into account as it is difficult to assume that a PSC officer
and/or sampling team member would stay on-board the vessel for longer time periods or
several days.
In conclusion, for representative sampling, it seems realistic that at least 2 random sequential
samples are taken. In contrast, sampling over the entire time of the ballast water discharge
would be very difficult especially in cases where a long sampling time is required, e.g., more
than 2 hours, up to several days. Another aspect is night time sampling, i.e., cargo operations
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are regularly conducted also in night shifts, but PSC officers and/or sampling team and/or
laboratory for analyses may only be available at day shifts.
Another challenge may become the need to obtain a representative sample of the whole
discharge of a vessel, when the vessel will be discharging ballast water from more than one
ballast water origin. For such cases it is recommended, if possible, that at least one
sequential sample per ballast water uptake source is taken. If a single tank was filled from
multiple sources this does not trigger necessity for two or more samples.
4.8 SAMPLING EQUIPMENT AND SAMPLING POINT
ARRANGEMENTS
Sampling equipment is meant to include all the equipment a PSC officer or sampling team
would need to bring onboard a vessel to conduct sampling for compliance. Sampling
arrangements include all the arrangements which would need to be setup on vessels to enable
sampling for compliance monitoring.
4.8.1 SAMPLING EQUIPMENT
It is clearly advisable to use light-weight and robust equipment of compact design to ease the
transport and use onboard a vessel. Sampling equipment presented here was tested and used in
onboard sampling studies and BWMS type approval tests, and is presented here only as an
example.
4.8.1.1 Plankton net for in-line sampling
For net tows through an opened ballast water tank, cone-shaped net have shown to sample the
water efficiently. The net diameter should be smaller than 30 cm and it should be shorter than
100 cm in length. The shape and dimensions of such a net reduces the risk that the net
becomes stuck inside the tank during sampling.
As a net mesh size it is recommended to use a mesh of 36 micrometres in the square
dimension, which results in a diagonal diameter of 50 micrometres. This is in line with G8, i.e.
If samples are concentrated for enumeration the samples should be concentrated using a sieve
no greater than 50 micrometres mesh in diagonal dimension.
The plankton net should be equipped with a removable cod-end, preferably with filtering
panels so that the sample can be concentrated effectively. A valve at the bottom of the cod-
end eases the concentrated sample to be extracted. Should multiple samples be taken, it is
beneficial that the filtering sieve of the cod-end can be replaced so that no organisms become
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stuck from one sample and could be erroneously added to another sample. An example is
given as Figure 3.
Figure 3 - The plankton net for in-line sampling with a removable cod-end (photo taken
during shipboard test of the Severn Trent de Nora BWMS).
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4.8.1.2 Wash bottle
To clean the net and to drain all organisms caught in the cod-end an unbreakable wash bottle
may be used. This wash bottle may also be used when emptying the cod-end content, i.e.
concentrated sample, into an unbreakable sample bottle to ensure that all organisms caught
are transferred into the sample bottle. An example is given as Figure 4.
Figure 4 - Wash bottle used to drain all organisms caught in the cod-end (photo taken during
shipboard test of the Severn Trent de Nora BWMS).
4.8.1.3 Plankton net for in-tank sampling
For in-tank sampling a short plankton net with a small diameter is beneficial that it can easily
be lowered through a manhole. Studies have shown that the conical top increases the sampling
performance and at the same time it reduces the risk that the net becomes stuck in the ballast
water tank. A removable cod-end helps to clean the net between sampling events of different
tanks so that an “organism contamination” between sampling events can be avoided. An
example is given as Figure 5.
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Figure 5 – Plankton net for in-tank sampling.
4.8.1.4 Flow meter
A calibrated flow meter should be used to enable an accurate reading of the water volume
filtered through the plankton net. A flow meter should show also the sampling flow rate,
which is important for appropriate sampling planning and setup Two examples are given as
Figure 6.
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Figure 6 - Flow meters are battery powered, the one on the left is intrinsically safe for use on
tankers (photo on the left taken during shipboard test of the Severn Trent de Nora BWMS).
4.8.1.5 Water Column Sampler
The water column sampler is of dimensions that allow entering the ballast tanks through
sounding pipes, but can be used also via manholes. While the water column sampler is being
lowered down the ballast water enters through the hole on the upper side.
The water column sampler was designed to sample a water column. The sampler is lowered
down into tank until the bottom is reached and than pulled back up. The water enters the
sampler through a 6 mm opening. The time to fill this sampler is approximately 10 seconds.
When lowering the water column sampler, water will be proportionally sampled from the
entire water column provided the sampler is lowered through the water column with a
constant speed. This may be achieved in 10 s. A maximum of 0.2 l of ballast water may be
sampled per one pull with this water column sampler. To increase the volume of water
sampled multiple replicates may be applied. A water column sampler example is given in
Figure 7.
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Figure 7 - Water column sampler.
4.8.1.6 Point-Source Sampler
The point-source sampler is of dimensions that allow entering the ballast tanks through the
sounding pipes, but can be used also via manholes. The point-source sampler is lowered down
the sounding pipe to the desired point of sampling and than the valve is opened by pulling the
second rope which needs to connected to the valve. Point-source sampler can be used also to
sample the ballast water and sediments at the bottom of the ballast tank, when this is simply
lowered to the bottom and the valve when touching the bottom opens automatically.
Ballast water (and sediment) enters this sampler through the valves’ 3 mm opening. The time
needed to fill-up the sampler is approximately 1 minute and 0.225 l of ballast water may be
sampled per one pull. Multiple replicates will be used to increase the volume of sampled
water. An example is given as Figure 8.
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Figure 8 - Point-source sampler used for sampling ballast water through the sounding pipe.
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4.8.1.7 Pumps
• Air-driven Well Pump
o The Air-driven Well Pump is a compressed air driven pump. It may be lowered
down the sounding pipe or through a manhole to the desired depth or to the
bottom of the ballast tank to pump up ballast water, including sediment sludge.
The use of the Air-driven Well Pump is depth independent and it can to pump
up ballast water from greater depths e.g., >30 meters.
4.8.1.7.1 Hand pump
The hand pump used in previous studies light-weight and of compact design. It is
approximately 30 cm long with a diameter of 5 cm. The hand pump may be used, without
priming with water, until a maximum pumping head of ca. 9 m. A hose should be used which
is prepared to resist under pressure (extra strong hose walls). Lower hose through manhole or
sounding pipe to the desired depth and pump the water up into a bucket or directly through a
filtering device. The water should be filtered to enable the transport of smaller volumes of
water. An example is given as Figure 9.
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Figure 9 - Hand pump used for ballast water sampling.
4.8.1.7.2 Air-driven Well Pump
The air-driven well pump was designed to be used ballast water sampling at a desired water
depth, to sample the water column and/or for sampling of ballast water and related sediments
at the bottom of a tank. Pumping may start when pressurized air as supplied on a vessel (5–7
bar) is connected to the pump. The pump is lowered to the desired water depth. The flow rate
of this pump is between 1.3 and 2.0 l/min. An example is given as Figure 10.
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Figure 10 - Air-driven well pump used for sampling via sounding pipe and manhole/tank
hatch.
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4.8.1.8 Bucket
A bucket of 10 litre capacity seems suitable for the sampling events. To avoid objects and
dust etc to be blown in during the sampling event and also to ease transport of the sample a
bucket with a lid is preferred. Further, a volume scale on the bucket is helpful to allow
readings of the water volume collected. An example is given as Figure 11.
Figure 11 - A 10 litre bucket with a volume scale (photo taken during shipboard test of the
Severn Trent de Nora BWMS).
4.8.2 IN-TANK SAMPLING ARRANGEMENTS ON VESSELS
There are no in-tank sampling points arranged on vessels. Ballast water may under certain
conditions be accessed via manholes, sounding pipes and air vents. The availability and
accessibility of these in-tank “sampling points” has shown to be critical.
Sampling point availability and accessibility are specific and dependent on ship type, design,
age, dimensions and current ship operations, e.g., cargo operations, restoration or repair works.
Three different general patterns were identified:
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• ships which do not carry cargo on the weather deck have easier access to sampling
points located on that deck. This is critical especially for the access to ballast tank
manholes;
• larger vessels have generally more suitable and accessible sampling points, e.g., due to
more space to place and operate the sampling equipment, wider sounding pipes;
• newer ships and those better maintained show easier sampling points access to the
ballast water because no or less rusty screws and nuts on manholes or venting pipes
need to be opened, no or less rust occurs inside sounding pipes etc.
Manholes are available on all ballast tanks. However, the experience from David & Perkovič
(2004) has shown that only 20% could be opened for sampling. Limitations observed included
very rusty screws and nuts (which would need to be cut-off to open the manhole cover), some
covers were cemented, or the manhole was cowered by cargo. In other cases the access was
limited because of ongoing cargo operations (80%). Venting pipes are also available on all
ballast tanks and were mostly accessible, but most of their covers are fixed with rusty nuts
thereby limiting the access for sampling. The most and easiest accessible sampling points on
all inspected vessels were sounding pipes.
As a next step during this study the requirements (rules) of some members of the International
Association of Classification Societies, London (IACS) regarding the construction of
sounding pipes have been analysed. The minimum sounding pipe requirements include:
• all ballast water tanks should have sounding pipes, which need to be as straight as
practicable,
• sounding pipes should not be less than 32 mm of internal diameter, and
• they must always be accessible (Lloyd’s Register of Shipping, 1996; Det Norske
Veritas, 2000; American Bureau of Shipping, 2001; Bureau Veritas, 2002;
Germanischer Lloyd, 2002).
When analysing the sampling accessibility of ballast water through this sampling point, some
technical limitations were also identified. These are the pipe diameter and the water depth
level inside the tank from the sampling point on deck, i.e., head. David & Perkovič (2004)
also noticed that most sounding pipes have a welding under the cover, which narrows the
access into the pipe by a reduced pipe diameter. Sampling equipment such as plankton nets,
traps, buckets etc. cannot be used for sounding pipe ballast water sampling. Further, suction
pumps are practically excluded if the pumping head is more than 9 m. Therefore, sounding
pipe sampling at greater depths to double bottom tanks, e.g., some ships may have only
double bottom tanks, or ballast water may just be carried in double bottom tanks, will require
a well pump of smaller diameter to be lowered down the pipe.
Should a pump be used, this should have a capacity to pump up water from greater depths and
at the same time to cause no damage to organisms. Several pumps are available, but were not
specifically designed for shipboard ballast water sampling. Pumps which require power
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supply, impose a limiting factor especially for their use on board vessels that transport oil and
oil products or different dangerous cargoes. To overcome this limitation pumps driven by
compressed air may be used, which is already available on almost all ships. The current
sounding pipes design allows the water and associated sediment only to enter from the bottom
end of the pipe, which was also recognized as a possible factor impacting representativeness
of sounding pipe samples.
4.8.3 IN- LINE SAMPLING ARRANGEMENTS ON VESSELS
To be able to sample from a ballast water discharge line, appropriate permanent sampling
arrangements need to be setup on the tested vessel in an area with enough space to safely
conduct a sampling event. The permanently installed sampling arrangements would include:
• sampling point installed in ballast water discharge line (see below);
• isokinetic sampling facility (see below),
• discharge point for the discharge of the sampled water installed in ballast water
discharge line, and
• hook or other installation that the plankton net can be hang ca. 100 cm directly over
the middle of the sampling bin, and
• discharge pump to empty sampling bin during sampling. The pump to pump out the
exciding water from the sampling bin after being filtered through the sampling net
should be of capacity to withstand head pressure in the ballast discharge line. It is also
important to have a valve which allows for regulation of discharge flow from the bin
to be able to provide for adequate level of water in the bin during sampling, i.e., after
achieving adequate level of water to have sampling net seating in the water
simultaneous discharge with sampling flow needs to be established.
Equipment for the temporary use at the sampling point, but which should be stored on the
vessel include:
• sampling bin used to place the net in water during sampling,
• valve at the discharge point or pump to manipulate the water level in the sampling bin,
• hoses to connect from the sampling point to the flow meter and plankton net as well as
from the sampling bin to the discharge point.
The sampling bin should be of big enough capacity that the plankton net can be placed into
the bin at best completely. There a bin of ca. 100 cm height and approximately 50 cm
diameter is needed. A bin of this size allows enough space to have the sampling net placed in
the water in the bin when filtering ballast water so that it is permanently submerged. This is
very important to avoid die-off of filtered organisms because of being exposed to different
stresses during filtration.
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Pump used for the discharge of sampled water should be of adequate capacity to enable
simultaneous sampling and discharge of sampled water from the bin.
Figure 12 - Elements of a sampling arrangement for in-line sampling on vessel.
FLOW
METER
INFLOW
OUTFLOW
~ 2-2.5 m
~ 1m
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Figure 13 - Sampling arrangement for in-line sampling on a vessel (photo taken during
shipboard test of the Severn Trent de Nora BWMS).
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It may be considered to permanently install a sampling arrangement so that the PSC only
needs to bring the “consumables” on board, such as a new plankton net, sample bottles,
buckets etc. Such sampling skids are currently being developed and tested with first tests on
commercial vessels (IMO 2008, 2012, Lemieux et al., 2010, Schillack 2013, Wier 2013).
However, rigorous on board validation tests are lacking.
4.8.4 SAMPLING POINT
Vessels are of very different sizes, design and arrangements, mainly depending on their
purpose and age. Consequently, also ballast water systems are of very different capacities and
designs. The G2 Guidelines suggest the installation of an isokinetic sampling point/facility,
which diameter is related to the diameter of the ballast discharge line where this is installed.
With this, a range of different sampling points would be expected on different vessels, posing
a real challenge to PSC to have adequate sampling equipment for all different sampling points.
Actually, sampled water goes from the sampling point through a sampling flow meter to
measure sampled quantity, and this would then also need to be of the same size. This looks to
be impractical.
Sampling experience for type approval shows that most vessels have installed 1 inch sampling
points. A sampling point of 1 inch has shown to deliver enough flow (between 45 and 90
litres/minute) for sampling purpose.
To enable sampling for compliance conducted by PSC, a vessel should have a sampling point
installed. The sampling point should have:
• enough space to safely conduct a sampling event,
• a sampling bin where water is collected and further pumped out,
• a pump to pump out the exciding water after being filtered through the sampling net.
Guideline G2 defines the “Sampling Facilities” as the equipment installed to take the sample
and the “Sampling Point” as that place in the ballast water piping where the sample is taken.
This means that the sampling point is the part of the vessel’s main pipe where the sampling
facility is installed. Guideline G2 provides further details for isokinetic sampling facilities:
In order to undertake an accurate measurement on the organism concentration in the ballast
water, it is recommended to install an “isokinetic” sampling facility. Isokinetic sampling is
intended for the sampling of water mixtures with secondary immiscible phases (i.e. sand or
oil) in which there are substantial density differentials. In such conditions, convergence and
divergence from sampling ports is of significant concern. Since most organisms are relatively
neutrally buoyant, true isokinetic sampling is unnecessary. However, the mathematics related
to isokinetic sampling are deemed to be useful as a basis for describing and specifying
sampling geometries. Isokinetic sampling is necessary to ensure that a sample contains the
same proportions of the various flowing constituents as the flow stream being sampled.
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During isokinetic sampling the sampling device does not alter the profile or velocity of the
flowing stream at the moment or point at which the sample is separated from the main flow
stream. Under isokinetic conditions, the velocities of both the sample and the main flow are
equal at the point at which the sample is separated from the main flow. To achieve isokinetic
sampling conditions, a sampler is designed to separate a subsection of the total flow-stream
in a manner that does not encourage or discourage water entry other than that which is
otherwise in the cross-section of the sampler opening. In other words, flow streams in the
main flow of the pipe should not diverge or converge as they approach the opening of the
sampler.
The sampling pipe should be elbow-shaped with the opening in the middle of the ships´ main
ballast water pipe directed towards the water flow in the line (see Figure 14).
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Figure 14 - Panel A, sampling port within a straight stretch of ballast line. Panel B, sampling
port at a 90 degree elbow in the ballast line. Arrows indicate direction of ballast flow.
4.9 SAMPLES HANDLING
As stated in the G2 Guidelines, samples should be transported, handled and stored with the
consideration of a chain of custody procedure.
Tape should be used to seal the sample lid to the sample bottle to avoid water leakage.
4.9.1 SAMPLES LABELLING
Each sample needs to be clearly labelled and the label secured that it cannot fell off. G2
recommends that each sample container should be labelled by, e.g., using a waterproof
permanent marker and additional paper which may be deposited inside the sample container.
The information recorded should include but not be limited to the date, ship name, sample
identification code, tank numbers and preservative(s) if used. Should the sample be
concentrated the original volume should be added to the label. This seems to be
comprehensive information to be written on a sample bottle and it is recommended that each
sample will clearly be numbered and essential information be written on the sample bottle.
Thereafter, additional information should be recorded on paper as a sample collection data
form (see G2) and the form and sample bottle be stored together in a sealed plastic bag.
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4.9.2 SAMPLES TRANSPORT
All samples should be transported in (Styrofoam) cooling boxes with several cooling elements.
Once the samples reached the lab, they should be stored at best at a comparable temperature
as the ballast water showed during sampling. The samples should be analysed as soon as
possible, but no later than 6 hours after sampling to avoid a die off of organisms.
Organisms greater than or equal to 50 micrometres in minimum dimension:
• The organisms sampled should not be concentrated below 1 l of water to enable
appropriate sample transport.
• After sampling the sample needs to be transferred into the cooling box as soon as
possible and the lid placed back on the cooling box to avoid light penetration or
heating up.
Organisms less than 50 micrometres in minimum dimension and greater than or equal to 10
micrometres in minimum dimension:
• The sample should not be concentrated. Three subsamples should be taken of not less
than 100 ml to enable appropriate sample transport.
• After sampling the sample needs to be transferred into the cooling box as soon as
possible and the lid placed back on the cooling box to avoid light penetration or
heating up.
Indicator microbes
• The water should not be concentrated. Three subsamples should be taken of not less
than 1 l to enable appropriate sample transport.
• The 1 l sample taken should after sampling be transferred into the cooling box as soon
as possible and the lid placed back on the cooling box to avoid light penetration or
heating up.
4.9.3 CHAIN OF CUSTODY
A sample transfer protocol should be completed and signed as a chain-of-custody procedure.
As stated in G2, the sample collection data form and chain of custody record should be kept
with each individual sample.
G2 further recommends that the following should be recorded in the chain of custody
- Information to contain a complete record of the persons handling the sample from the time of
the sampling onwards.
- Date, ship identification, sample identification code, and a list of people who have handled the
sample, including the person who takes the sample, dates and time, and the reason for sample
transfer and the integrity of the sample on transfer.
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5 DISCUSSION
Ballast water sampling may be conducted for different aims: to assess the biology and
chemistry of ballast water (scientific research); to identify potentially harmful or other
organisms carried in ballast water (risk assessment); and, to assess compliance with ballast
water management requirements (monitoring and enforcement) which is the focus of this
contribution. Ballast water sampling, is complex due to differences in organisms' dimensions
and behaviour, as well as to differences in ship construction including the availability of
sampling points. These issues as well as the aims of the ballast water sampling study impact
the sampling method selection.
The sampling point is clearly related to the sampling goal, e.g., indicative, D-1 or D-2
standards compliance sampling. The in-line sampling point will need to be installed on
vessels according to the G8 Guidelines, but there are no provisions for in-tank sampling
points. Therefore, ballast water to be sampled from a tank needs to be accessed via existing
openings, i.e., manholes, sounding or air pipes. The availability of these has proven to be a
critical issue. Employment of sampling equipment, modified for on board use, and a flexible
approach is needed to allow sampling via the different access points.
New methods were developed to ease ballast water sampling on board ships including
especially designed equipment for in-tank sampling through sounding pipes. Sounding pipe
sampling was achieved by the use of an air-driven well pump, hand pump, a water-column
sampler, and a bottom and sediment sampler. On board tests have shown that this sampling
equipment may be used to sample most target organisms despite some size limitations may
occur according to the opening dimension of the sampling tool. The tests also confirmed that
all three water samplers can be safely used on almost all ships, while not disturbing standard
ship operations conducted in the port. In addition to their deployment via sounding pipes these
especially designed samplers may also be employed via manholes or tank hatches.
In-tank sampling may be more appropriate for scientific research and risk assessment with the
aim to assess ballast biota, while at discharge sampling is more appropriate for the monitoring
of compliance with ballast water management requirements. However, certain tanks are not
discharged through pipework on board, but may use gravity to empty them. In those cases in-
tank sampling is the only way to prove compliance with ballast water management standards.
Further, in-tank sampling may also be used to document risk assessment results, e.g., to proof
the presence or absence of target organisms before the ballast water is being discharged.
Sampling for zooplankton via the sounding pipes does not result in a representative sample of
species in the tank as comparisons of sounding pipe and manholes samples from the same
tank found that net samples were more diverse. Sounding pipe samples contained 0-60% of
the organisms of a net sample indicating the need to sample ballast tanks via opened man
holes. Further, pumps used via open manholes delivered more diverse samples than net
samples, therefore pumps may be considered when the sampling via manholes. Future ballast
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water studies should take into account that sampling via sounding pipes is inferior when
selecting appropriate sampling techniques. However, in many cases manholes cannot be
opened due to, e.g., overlaying cargo or cargo operations in the area where the manhole is
located, and in these instances sounding pipe sampling might be the only solution to sample
the ballast water at all.
If the sampling is to document non-compliance, i.e., violation of the ballast water discharge
standard, much less onerous sampling requirements are posed to the port state as a
demonstration that an explicit value is exceeded. For example, should a sample from the tank
contain 1000 living organisms greater than 50 micrometres in minimum dimension and the
tank capacity is 100 cubic metres, the organism concentration would exceed the D-2 standard
when the water is discharged.
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6 CONCLUSIONS AND RECOMMENDATIONS
Many different ballast water sampling methods and equipment have been used for different
purposes. Shipboard sampling is also conducted for BWMS testing for type approval. Hence,
shipboard sampling methods for testing BWMS actually exist, and these have been approved
by different national responsible authorities. However, studies have shown that sampling
results may be biased by different sampling processes because of, e.g., patchy distribution of
organisms in tanks, die-off of organisms during sampling etc. As there is still no commonly
agreed ballast water sampling methodology or approach, this may impact representative
ballast water sampling, and certain vessels may be found in compliance in one port, but not in
another.
Ballast water sampling studies have shown that different methods and sampling equipment
may be used for different sampling goals, e.g., D-1 or D-2 standards, indicative or detailed
sampling. Sampling methods and equipment also depend on ballast water access points, i.e.
in-tank via manholes, sounding pipes or air vents, or in-line installed sampling points, and on
the target groups of organisms, i.e., organisms greater than or equal to 50 micrometres in
minimum dimension, organisms less than 50 micrometres and greater than or equal to 10
micrometres in minimum dimension, and indicator microbes.
For this contribution it was of prime importance to consider the appropriate sampling
approach for compliance control according to the BWM Convention. Ballast water sampling
methods suggested here for compliance with the D-2 standard (detailed sampling) and the
indicative test (indicative sampling) are based on the sampling experience gained on more
than 60 shipboard tests for type approval of 15 different BWMS. Very importantly, these
sampling methods were also scientifically tested, improved and validated during three studies
on representative BWS for compliance monitoring, conducted for the Federal Maritime and
Hydrographic Agency, Hamburg, Germany in 2009 and 2012, and for the European Maritime
Safety Agency, Lisbon, Portugal in 2010.
For compliance with the D-2 standard it is suggested that samples should be taken during
discharge, i.e., from the ballast water discharge line after the pump prior to the discharge. This
approach delivers the most accurate results of organisms being discharged in ballast water
from a vessel as a side stream of the discharge. Samples should be taken in two or more 10
minute sampling sequences, sampling 300-500 litres in each sequence. With this, organisms
are less exposed to die-off during sampling, such sampling is logistically more appropriate
than sampling over the entire discharge time, and also scientific studies have shown this
sampling method to be representative of the whole discharge. In reality, this sampling method
is not exactly the same as for the type approval testing of BWMS, but has all components of it,
e.g., the same sampling equipment may be used, and in two or three sequences PSC may
sample more than 1000 litres. This method is also in-line with the agreement at the IMO that
sampling methods applied by PSC for compliance monitoring, i.e., G2 sampling, should be no
more stringent than the methods applied for BWMS type approval, i.e., G8 sampling.
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However, one key problem remains with in-line (at discharge) sampling approaches and this
is that compliance or non-compliance can only be proven while the ballast water is being
pumped overboard. Consequently, the potentially non-compliant ballast water may already
have been released before it is clear whether or not it is in compliance with the BWM
Convention standards. Should high risk organisms be suspected in the ballast water to be
discharged, in-tank compliance control sampling may be more appropriate compared to the
in-line approach to possibly avoid discharge of not properly treated ballast water. In-tank
sampling may also be the only approach possible if the ballast tanks to be discharged have
only direct discharge to the sea, hence in-tank sampling remains important to be dealt with.
This study presents also different in-tank sampling methods using different sampling
equipment, however their efficiency would need to be studied more in detail, scientifically
validated, and cross compared. Nevertheless, if the in-tank sampling is to document non-
compliance, i.e., violation of the ballast water discharge standard, much less onerous sampling
requirements may be posed to the port state as a demonstration that an explicit value is
exceeded. For example, should a sample from the tank contain 1000 living organisms greater
than 50 micrometres in minimum dimension and the tank capacity is 100 cubic metres, the
organism concentration would exceed the D-2 standard when the water is discharged.
At the last BLG 17, the BWS Guidance in a form of Circular was finalised. This IMO BWS
guidance provides the current state of knowledge of methods and approaches for ballast water
sampling and analysis, which were presented at IMO. It was recognized that many of the
sampling and test methods in the BWS Guidance have not yet been adequately validated, and
have not yet been fully integrated in port State control procedures in order to validate their
practical utility for determining compliance with the BWM Convention. Given that these
methods are rapidly improving, members and observers were encouraged to further develop
sampling and analysis protocols, including but not limited to, the range of options outlined in
the BWS Guidance.
Further it was suggested that once the BWM Convention enters into force, a trial period of 2
to 3 years would be initiated where PSC can further trial the approaches in the BWS Guidance
to ensure they are practical and fit for purpose. During the trial period it is anticipated that
port States share the results of sampling and analysis. After the first trial year results and
findings from sampling efforts should be communicated to IMO. This will likely result in an
update of the BWS Guidance. After the second trial year this will be repeated to determine if
changes are needed to standardize the options available. Lastly, after the third year
recommendations are provided to MEPC on standardized sampling and analysis protocols by
advances in scientific knowledge being communicated to IMO which may be considered to
update the BWS circular accordingly.
This study has shown that there are BWS methods which were extensively used on board
vessels to test BWMS to proof compliance especially with the D-2 standard, and were
scientifically validated by additional tests and studies. These BWS methods have also shown
to be relatively simple, i.e., no special background education is needed for their application,
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they are cost effective, i.e., there is no need for very expensive equipment, and there are no
high running costs, and they are generally applicable on all vessel types and in all geographic
regions. The sampling recommendations suggested in this study therefore may result in a
workable, equitable and pragmatic solution to support the entry into force of the BWM
Convention.
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7 ACKNOWLEDGEMENTS
The author thanks several funding agencies for having enabled the ship sampling studies.
Ballast water sampling experience referred to in this report was generated during the
following research projects:
(a) "Harmful Introductions and Ballast Water Management in the Slovenian Sea", financially
supported by the Ministry of Education, Science and Sports of the Republic of Slovenia and
the Port of Koper (Luka Koper d.d.),
(b) onboard ballast water sampling voyage "Ship-board ballast water sampling trials to take
representative samples for compliance control with the D-2 Standard of the Ballast Water
Management Convention" funded by Federal Maritime and Hydrographic Agency, Hamburg,
Germany,
(c) onboard ballast water sampling study “Testing Sample Representativeness of a Ballast
Water Discharge and developing methods for Indicative Analysis” funded by the European
Maritime Safety Agency (EMSA), Lisbon, Portugal, and
(d) onboard ballast water sampling study “Recommendations for Representative Ballast
Water Sampling” funded by Federal Maritime and Hydrographic Agency, Hamburg,
Germany.
Grateful thanks for all support and help provided during shipboard tests are also expressed to
all vessel crews, shipping agents, port State control personnel and ship operators, which are
too numerous to be mentioned here individually.
For analysis of the phytoplankton samples during these shipping studies I thank the expert
team of the Royal Netherlands Institute for Sea Research (NIOZ), Texel, the Netherlands, and
especially Alex Blin and Louis Peperzak.
I further like to thank Stephan Gollasch, GoConsult, Germany for his critical review of this
document.
Last, but not least, WWF International is thanked for having provided funds to generate this
report.
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LIST OF FIGURES
Figure 1 - Annual world ocean´s surface salinity (Source: http://www.ccpo.odu.edu/
~arnoldo/ocean405/s00anw01.gif). .......................................................................................... 14
Figure 2 - Ballast water discharge above pier level from the upper wing tanks of a bulk carrier
(Jure Barovic, with courtesy of the Port of Koper, services for protection of the sea). ........... 19
Figure 3 - The plankton net for in-line sampling with a removable cod-end. .......................... 32
Figure 4 - Wash bottle used to drain all organisms caught in the cod-end. ............................. 33
Figure 5 – Plankton net for in-tank sampling. .......................................................................... 34
Figure 6 - Flow meters are battery powered, the one on the left is intrinsically safe for use on
tankers. ..................................................................................................................................... 35
Figure 7 - Water column sampler. ............................................................................................ 36
Figure 8 - Point-source sampler used for sampling ballast water through the sounding pipe. 37
Figure 9 - Hand pump used for ballast water sampling. .......................................................... 39
Figure 10 - Air-driven well pump used for sampling via sounding pipe and manhole/tank
hatch. ........................................................................................................................................ 40
Figure 11 - A 10 litre bucket with a volume scale. .................................................................. 41
Figure 12 - Elements of a sampling arrangement for in-line sampling on vessel. .................. 44
Figure 13 - Sampling arrangement for in-line sampling on a vessel. ...................................... 45
Figure 14 - Panel A, sampling port within a straight stretch of ballast line. Panel B, sampling
port at a 90 degree elbow in the ballast line. Arrows indicate direction of ballast flow. ........ 48
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LIST OF TABLES
Table 1 - Possible sampling access points, equipment and other details recommended for
compliance control sampling with the D-1 standard. ............................................................... 16
Table 2 - In-line sampling equipment and other details recommended for compliance control
sampling with the D-1 standard. .............................................................................................. 18
Table 3 - Possible sampling access points, equipment and other details recommended for
indicative compliance control in-tank sampling with the D-2 standard. .................................. 22
Table 4 - Possible sampling access points, equipment and other details recommended for
indicative in-line compliance control sampling with the D-2 standard. ................................... 25
Table 5 - Possible sampling access points, equipment and other details recommended for
compliance control sampling with the D-2 standard. ............................................................... 27
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