Kaag, N.H.B.M.

Sneekes, A.C.

English

Name not available

The Road from Approval to Compliance

Wageningen Yield 

 P012-1 
The Road from Approval to Compliance 
 
A.C. Sneekes & N.H.B.M. Kaag 
 
IMARES Wageningen UR, Dept. Environment. P.O.Box 57, 1780 AB  
Den Helder, The Netherlands. Email: andrea.sneekes@wur.nl 
 
Abstract 
 
Ballast water treatment systems are faced with two contrasting challenges; the 
treatment needs to be very efficient in removing or killing organisms on the one hand, but 
should not pose an environmental risk at discharge on the other. At this moment a suite of 
techniques is used to assess the efficacy and the environmental risk of ballast water 
treatment systems and improvements are still going on. No single technique is sufficient 
on its own, but using a combination of techniques facilitates a “weight-of-evidence” 
approach. 
 
The next challenge in controlling ballast water is compliance monitoring that should 
allow port authorities to determine the efficacy of a treatment and judge the 
environmental risk at discharge. Using the full battery of testing techniques for each 
system and at each discharge will be practically impossible. Therefore we propose to 
select a deliberate set of testing techniques on the basis of the information compiled 
during land-based and toxicity testing. Examples of such an approach will be presented 
based on our ballast water experiences in the last five years, but also many years of field 
monitoring.  
 
Key words: Ballast Water, Efficacy, Environmental risk, Compliance Monitoring, 
Bioassays, Biological Early Warning Systems, Rapid Screening Assays. 
 
1 Introduction 
In 2004, the International Convention for the Control and Management of Ship‟s 
Ballast Water and Sediment was adopted. The goal of the convention was to reduce the 
number and rate of invasions of species outside their native range. This boosted the 
research and development of Ballast Water Management Systems (BWMS). The 
convention will not enter into force until ratification by 30 states representing 35% of 
merchant shipping tonnage. At 30 June 2010, already 26 States representing over 24% of 
world tonnage have signed the convention which makes the ratification getting closer and 
closer. Signed parties are, however, already obligated to prevent, minimize and ultimately 
eliminate the transfer of harmful aquatic organisms and pathogens through the control 
and management of ships‟ ballast water and sediments.  
 
For the registration procedure, IMO has set out standards for development of BWMS, 
and new systems must be approved by the Administration in accordance with IMO 
Guidelines. Only looking at the environmental part, BWMS must be rigorously tested 
proving the efficacy of the system without causing greater harm than they prevent to their 
environment. Although both types of analyses have the same biological background, the 
endpoints are markedly different: efficacy aims 100% effect, environmental risk 
prevention aims no effects. 
 
The development of Ballast Water Management Systems plus monitoring of the 
efficacy of the treatment without causing negative effects to the environment is a great 
challenge for technicians, chemists, biologists, but also policy makers. As the date of 
 P012-2 
ratification is getting closer, the pressure on port state authorities is rising. They pose the 
challenge in monitoring all BWMS that passed the IMO guidelines and enter their 
harbour for ballasting and/or de-ballasting on bases of efficacy, but also determining the 
lack of residual environmental risk.  
 
This paper points out some challenges on bases of our ballast water experiences for the 
approval procedures in the past five years with regards to efficacy testing and (residual) 
environmental risks. And an ecotoxicological view on compliance monitoring built up 
from many years of field monitoring, specifically for the determination of the residual 
environmental risk.  
 
2 Approval procedure 
The approval procedure focuses on the main goal: preventing, minimizing and 
ultimately eliminating the transfer of harmful aquatic organisms and pathogens. For the 
Administration, guidelines for approval of a BWMS are described in G8. Testing the 
efficacy of the system by means of ship-board and land-based testing is a main part of 
these guidelines. Systems that make use of an active substance must also research the 
sustainable use of the substances concerning ship safety, human health and the aquatic 
environment as described in procedure G9.  
 
2.1 Efficacy Testing 
The Ballast Water Performance Standard (Regulation D-2) stipulates that discharged 
ballast water shall contain less than 10 viable organisms per cubic metre greater than or 
equal to 50 micrometres in minimum dimension and less than 10 viable organisms per 
millilitre less than 50 micrometres in minimum dimension and greater than or equal to 10 
micrometres in minimum dimension; and discharge of the indicator microbes shall not 
exceed the specified concentrations.  
 
The IMO criteria do not distinguish between survival and mortality, but between viable 
and non-viable. It seems that makes life easier, as organisms do not have to be killed. The 
only objective is that reproduction is made impossible. Determination of viability is, 
however, a complicated assessment. Mortality -how difficult it may sometimes be to 
assess unequivocally- is rather instantaneous. Viability needs long-term analyses, if ever 
it can be assessed beyond doubt. 
 
An example of this was seen during the land-based tests of the ErmaFirst BWMS at the 
NIOZ test facilities. It appeared that barnacle cyprid larvae were not killed by the 
chlorine that was used as active substance. A laboratory test with different doses of 
hypochlorite showed that cyprid larvae of Balanus amphitrite were not killed below 10 
mg/l free chlorine and even at 100 mg/l still surviving larvae were observed (Fig. 1). 
However, settlement of the cyprid larvae did not occur at 6 mg/l and higher. This means 
they are not viable, as the cyprid larva itself is a non-feeding stage which will eventually 
die of starvation when not settled. A 3-day test like this is of course no definitive proof, 
but also in the land-based test, no settlement was observed in the treated tanks (Kaag & 
Sneekes, 2010). 
 
 P012-3 
0.1 1 10 100
0
25
50
75
100 Survival
Settlement
concentration free chlorine (mg/l Cl2)
p
e
rc
e
n
ta
g
e
 
Fig. 1 Graphical view of the results from the laboratory barnacle settlement test 
 
Similarly, organisms can be temporarily disabled by a treatment. This may give the 
impression that they are dead, but after recovery they may appear to be very viable. For 
instance, when phytoplankton is treated with low doses chlorine, staining with neutral red 
reveals no viable cells during the first 24h. This is supported by measurements of 
chlorophyll-a and photosynthetic activity. However, within a few days the phytoplankton 
starts growing again vigorously in re-growth experiments. Extrapolation of the growth 
curves back to the treatment day indicate that the number of viable cells must have been 
much higher than is acceptable (1000-2000 cells/ml). 
 
2.2 Environmental Risk 
IMO obligates Parties not to cause greater harm than they prevent to their environment. 
In an ideal situation, all species available in the environment are screened for their 
sensitivity to the discharge water from a BWMS. As this is not possible, risk assessment 
relies on the representativeness of a group of test-species. BWMS that make use of an 
active ingredient must prove with means of biological toxicity tests, the so-called 
bioassays that there is no significant negative effect of ballast water discharge to the 
receiving environment. We have a suit of standardized bioassays available as shown in 
Table 1 that covers, but is not limited to, the three main groups used in risk assessment: 
Algae, Crustacean and Fish. 
 
Table 1 Typical standardized bioassays used in Ballast Water Research by IMARES 
Wageningen UR, the Netherlands. 
Group Species name Application 
Bacteria Vibrio fischeri  fresh to saltwater 
Algae Phaeodactylum tricornutum brackish-saltwater 
 Pseudokirchneriella subcapitata freshwater 
Crustacea Artemia franciscana brackish-saltwater 
 Acartia tonsa saltwater 
 Daphnia magna freshwater 
Mollusc Crassostrea gigas saltwater 
Rotifer Brachionus plicatilis brackish-saltwater 
 Brachionus calyciflorus freshwater 
Fish Solea solea  saltwater 
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A wide range of bio-assays and species should be evaluated for the assessment of 
environmental risk. Our own experience not only with ballast water tests, but also with 
marine and freshwater effluents, single substances, concentrated extracts and sediments, 
as well as with numerous multi-species mesocosm experiments, shows that no single 
species or species group is the „most sensitive‟ for all substances. For that reason an 
„intelligent‟ choice of bioassays is necessary.  
 
Some indication regarding the most sensitive species may be extracted from literature 
and specific database (e.g. Aquire, EUCLID). It has to be considered, however, that most 
bioassays were developed for use in standard laboratory conditions in order to derive the 
dose-response for single substances. Treated ballast water, however, should be 
considered an effluent and needs another method of approach. The composition of the 
water to be tested is sometimes far from standard and specific water characteristics often 
are out of the specifications for the specific test. Although these are a result of the 
treatment, they may obscure effects of the active substance itself and manipulation of the 
water characteristics may be needed, as well as selection of other test species.  
 
To conclude, no single assessment is sufficient to determine either the efficacy of a 
BWMS, or the safe concentration of ballast water at discharge. It is the combination of 
chemical analysis of active substances, different methods for viability analysis, a suit of 
bioassays and literature/database data that together give a picture of performance of a 
BWMS and potential environmental risks. In ecotoxicology this is called a „weight-of-
evidence‟ approach. 
 
3 Compliance Monitoring 
Compliance monitoring aims at two questions: has the treatment been sufficient and -if 
active substances are used- does the discharged water pose a risk to the receiving 
environment.  
 
Most BWMS are tested under controlled circumstances and experts have tested the 
water quality rigorously according to IMO criteria. Some discrepancies in the guidelines 
must be acknowledged and raise new challenges for monitoring. For example, the 
guideline asks to test a BWMS at two different salinities. When chosen for two high 
salinities (>20 psu), this might not reveal any problems, but when tested at low salinity 
(freshwater conditions) problems with the efficacy or residual toxicity of the system 
might occur. Some systems need a certain composition of the water in order to produce 
the active substance. A minimum salinity is for instance needed to produce sufficient 
chloride by electrolysis. This may be achieved in freshwater by adding brine, although 
this changes the basic characteristics of the water.  
 
Another challenge can be found in temperature. Most often a system is tested in one 
continent under favourable conditions with respect to temperature and biology, the so-
called biological season, in order to meet the specified criteria for inlet water. 
Physical/chemical reactions differ with different temperatures and can result in for 
instance insufficient degradation of the compound for a short trip, showing toxicity at 
discharge. But also, organisms found in winter time compared to summer time can react 
differently to the BWMS. Often, resting stages of organisms are more robust as they need 
to survive less favourable conditions.  
 
3.1 Efficacy Testing 
Testing the efficacy of the system faces the same problems as we have seen in the 
registration procedure: how to sample and recognize viable organisms. We will not go 
into detail on this subject, except for the following observation. Research on board of 
 P012-5 
ships, as has been presented on several occasions by Mr. Gollasch of GoConsult (e.g. 
Gollasch & David, 2010), has shown that the number of organisms may vary heavily 
between successive samples taken. Statistical analysis suggests that numerous samples 
have to be taken in order to obtain statistically sound results. On the other hand, Mr. Fuhr 
of NIOZ (Fuhr et al., 2010) suggested that the three samples should be taken immediately 
when discharge starts, as the analyses are very time consuming. Our own results of viable 
organism counts in three successive samples taken in-line during intake and discharge of 
treated ballast water give some practical nuance to this. At discharge, the treated water 
had been stored for 5 days in two 100 m3 tanks in a ship. For organisms <50 micron, as 
well as organisms >50 micron, the numbers varied less than one order of magnitude 
between replicates. Also, in our data, either all three samples were in compliance, or none 
of them was. This might suggest that one sample taken when discharge starts could give 
indication to judge whether to further investigate by sampling or not. 
 
3.2 Environmental Risk 
The question is whether active substances in the discharged ballast water still pose a 
risk to the receiving environment. Chemical analysis may be used to assess the 
concentration of an active substance in the ballast water before discharge. Reliable 
chemical analyses, however, usually take some time and not every chemical laboratory is 
able to analyze all potential active substances. For some substances in-line monitoring of 
the concentration is used as part of the BWMS and can be used as first indication. 
Laboratory analyses may be used as a way to check on these systems. An alternative may 
be the use of rapid test-kits which are available for many substances. These kits do have 
their limitations with regard to precision, but are very quick. The main problem with 
many of these kits is that their detection limits are often much higher than stated in the 
specifications due to the composition of the water. It may, therefore, not always be 
possible to measure as low as the safe concentration determined for the BWMS. 
 
Bioassays are useful alternative for the assessment of residual toxicity in ballast water. 
Unlike the situation described for land-based testing, there is no need to apply a whole 
battery of tests. Based upon the information taken up in the applications for Final 
Approval for each active substance cq. BWMS, the most sensitive species can be 
selected. It is, therefore, recommended to make this information accessible to port 
authorities. 
 
Algae quite often belong to the most sensitive species, which would make the algae 
growth inhibition test a suitable generic bioassay for compliance monitoring. This 
bioassay, however, lasts 3 days which is not very practical when a direct decision on the 
acceptability of the ballast water is needed. The same is true for most other bioassays. 
The typical test duration for ecotoxicological tests in 24h to 96h. The Microtox® Basic 
Test, using the bacteria Vibrio fisheri and comparable systems, may give results within 
one hour. Unfortunately, this test is often not sufficiently sensitive.  
 
The way to move on here is to explore the possibilities of Biological Early Warning 
Systems (BEWS) and other rapid on-line screening assays. Several BEWS are routinely 
used for monitoring the quality of surface water that is used as source for drinking water. 
Others BEWS, marine as well as freshwater, are or have been used for the on-line 
monitoring of water quality near calamities (spills), or sewage/waste water discharge 
points (Sneekes et al., 2005; Butterworth et al. 2001). Most BEWS are based on changes 
in behaviour in response of organisms to a chemical cue. This change is registered and an 
alarm triggered if a pre-set threshold (No. of animals responding, duration of response) is 
exceeded. Some experience of the operator is needed, as an alarm may also be triggered 
by the change of water characteristics (from holding water to ballast water for instance), 
 P012-6 
sudden noises, etc. Usually, these alarms can readily be distinguished from chemical cues 
by the duration of the response and the pattern of recovery. Some additional research is 
needed to explore the applicability of BEWS, especially with regards to ease of use, 
sensitivity to different active substances used, detection limits and whether different 
BEWS can be interchanged. 
 
Other potentially useful methods are rapid screening assays that are based upon 
standard bioassays. An example is the algae PAM-test. This test does not measure growth 
(inhibition) after several generations, but the direct inhibition of the photosynthetic 
system of the algal cells. In ecotoxicological testing, often an exposure period of 4h is 
used. In our tests with ballast water we have seen that a sensitive species may show a 
very pronounced response (to a high concentration) within 30 min. 
 
In the end, it is not up to us to decide how to perform compliance monitoring. This is a 
policy issue. We hope that this contribution gives some information that may help to 
make such decisions and we are always willing to discuss and clarify specific issues 
addressed. If, however, we have succeeded in clarifying some obscure issues, we will 
now blow some new smoke: 
 
The whole discussion on environmental risk and residual toxicity focuses on 
discharged ballast water. But what about toxicity of the water taken in? The water in most 
industrial harbour areas is not known for its ecological quality. This issue surfaced this 
autumn when we encountered severe toxic effects of the untreated ballast water control. 
This made an assessment of the success of the neutralisation impossible. In compliance 
monitoring there is no untreated control and the effect may be unduly attributed to the 
BWMS used. And even if it is clear that it originates from intake, would you allow 
discharge anyway? 
 
 
References 
 
Butterworth F.M., A. Gunatilaka & M.E. Gonsebatt (eds.) (2001): Biomonitors and 
biomarkers as indicators of environmental change 2. A handbook. Kluwer Academic/ 
Plenum Publishers, New York. 
 
Fuhr F., J. Finke, P.P. Stehouwer, S. Oosterhuis & M. Veldhuis (2010): Factors influencing 
Organisms Counts in Ballast Water Samples and Their Implications. In: Bellefontaine N., 
F. Haag, O. Linden, J. Matheickal (eds.) (2010): proceedings of the IMO-WMU Research 
and Development Forum. 26-29 January 2010. Malmö, Sweden. 
 
Gollash S., M. David (2010): Recommendations how to Take a Representative Ballast 
Water Sample. In: Bellefontaine N., F. Haag, O. Linden, J. Matheickal (eds.) (2010): 
proceedings of the IMO-WMU Research and Development Forum. 26-29 January 2010. 
Malmö, Sweden. 
 
Kaag N.B.H.M., A.C. Sneekes (2010): The sensitivity of barnacle cyprid larvae to chlorine. 
IMARES report C103/10. 
 
Sneekes A.C., W. Koops, N.H.B.M. Kaag, V.G. Blankendaal (2005): Response to harmful 
substances other than oil spilled at sea (HASREP). Task 3a: Investigation of the use of a 
flow-through musselmonitor® for monitoring incidental chemical releases at sea. TNO-
report R 2005/342. 


Biomonitors and biomarkers as indicators of environmental change 2. A handbook.

Blankendaal (2005): Response to harmful substances other than oil spilled at sea (HASREP). Task 3a: Investigation of the use of a flow-through musselmonitor® for monitoring incidental chemical releases at sea. TNOreport R

Recommendations how to Take a Representative Ballast Water Sample. In:

Sneekes (2010): The sensitivity of barnacle cyprid larvae to chlorine. IMARES report C103/10.

Veldhuis (2010): Factors influencing Organisms Counts in Ballast Water Samples and Their Implications. In:

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