SIST EN 17218:2019
(Main)Water quality - Guidance on sampling of mesozooplankton from marine and brackish water using mesh
Water quality - Guidance on sampling of mesozooplankton from marine and brackish water using mesh
This document specifies a method for the sampling of mesozooplankton from marine and brackish waters using mesh.
Wasserbeschaffenheit - Anleitung zur Probenahme von Mesozooplankton aus marinen und Übergangsgewässern mittels Netzen
Dieses Dokument legt Verfahren zur Probenahme von Mesozooplankton mithilfe von Netzen und kontinuierlichen Probenahmegeräten in marinen und brackigen Gewässern zum Zwecke der Bewertung der Wasserqualität und der Bestimmung des ökologischen Zustands von Ökosystemen fest.
Es wird eine Anleitung zu Probenahmeverfahren und den anschließenden Schritten der Konservierung und Lagerung gegeben. Die Probenahmeverfahren ermöglichen eine Abschätzung des Vorkommens von Arten und deren Abundanz (relativ oder absolut), einschließlich der räumlichen Verteilung und saisonaler sowie Langzeittrends für einen bestimmten Wasserkörper.
Die beschriebenen Verfahren gelten nur für die Probenahme von Mesozooplankton, das marine und brackige Gewässer bewohnt, und berücksichtigen nicht die flachen Uferzonen, die eine andere Art der Probenahme erfordern (z. B. Zooplankton in Salzmarschen).
Qualité de l'eau - Document d'orientation pour l'échantillonnage du mésozooplancton dans les eaux de mer ou saumâtres à l'aide de filets
Le présent document fournit des procédures pour l'échantillonnage du mésozooplancton dans les eaux marines et saumâtres, à l'aide de filets et de dispositifs d'échantillonnage en continu sur bandes collectrices, dans le but d'évaluer la qualité de l'eau et de déterminer l'état écologique des écosystèmes.
Des recommandations relatives aux procédures d'échantillonnage et aux étapes suivantes de conservation et de stockage sont fournies. Les procédures d'échantillonnage donnent une estimation de l'occurrence des espèces et de leur abondance (relative ou absolue), y compris la distribution spatiale et les tendances temporelles saisonnières et à long terme pour une masse d'eau donnée.
Les méthodes décrites sont limitées à l'échantillonnage du mésozooplancton qui habite les eaux marines et saumâtres et excluent les zones littorales peu profondes qui requièrent un autre type d'échantillonnage (par exemple, le zooplancton des marais salants).
Kakovost vode - Navodilo za vzorčenje mezozooplanktona v morskih in brakičnih vodah s pomočjo mrež
Ta dokument določa metodo za vzorčenje mezozooplanktona iz morskih in brakičnih vod s pomočjo mrež.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 17218:2019
01-julij-2019
Kakovost vode - Navodilo za vzorčenje mezozooplanktona v morskih in brakičnih
vodah s pomočjo mrež
Water quality - Guidance on sampling of mesozooplankton from marine and brackish
water using mesh
Wasserbeschaffenheit - Anleitung zur Probenahme von Mesozooplankton aus marinen
und Übergangsgewässern mittels Netzen
Qualité de l'eau - Document d'orientation pour l'échantillonnage du mésozooplancton
dans les eaux de mer ou saumâtres à l'aide de filets
Ta slovenski standard je istoveten z: EN 17218:2019
ICS:
13.060.10 Voda iz naravnih virov Water of natural resources
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
SIST EN 17218:2019 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 17218:2019
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SIST EN 17218:2019
EN 17218
EUROPEAN STANDARD
NORME EUROPÉENNE
May 2019
EUROPÄISCHE NORM
ICS 13.060.70
English Version
Water quality - Guidance on sampling of mesozooplankton
from marine and brackish water using mesh
Qualité de l'eau - Document d'orientation pour Wasserbeschaffenheit - Anleitung zur Probenahme von
l'échantillonnage du mésozooplancton dans les eaux de Mesozooplankton aus marinen und
mer ou saumâtres à l'aide de filets Übergangsgewässern mittels Netzen
This European Standard was approved by CEN on 15 March 2019.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17218:2019 E
worldwide for CEN national Members.
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EN 17218:2019 (E)
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 7
5 Sampling device . 8
5.1 General . 8
5.2 Nets . 8
5.3 Other field equipment . 10
5.4 Preserving solutions and other chemicals . 11
6 Prearrangements of sampling . 12
6.1 Documentation of strategies and methods . 12
6.2 Preparation of sampling equipment . 12
6.3 Safety instructions . 12
7 Sampling procedure . 12
7.1 Investigation programme . 12
7.2 Number and location of sampling sites . 13
7.2.1 General . 13
7.3 Diurnal sampling period . 14
7.3.1 General . 14
7.3.2 Sample size . 14
7.3.3 Geographical localization of sampling sites . 14
7.4 Operating the sampling device . 15
7.4.1 Vertical net hauls . 15
7.4.2 Horizontal tows/hauls . 15
7.4.3 Filling and labelling of sample bottles . 16
7.4.4 Preservation and storage of samples . 17
7.5 Field data recording . 18
8 Quality assurance . 18
Annex A (informative) Examples of sampling devices . 19
A.1 Bongo nets . 19
A.2 Continuous plankton recorder . 19
A.3 WP2 net . 20
A.4 Multinets . 20
A.5 Gulf VII sampler . 21
Annex B (informative) Preservation . 22
B.1 Preservation. 22
B.2 Formaldehyde (formalin) . 22
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B.2.1 General . 22
B.2.2 Advantages of formaldehyde . 22
B.2.3 Disadvantages of formaldehyde . 23
B.3 Lugol’s Iodine . 23
B.3.1 General . 23
B.3.2 Advantages of Lugol’s Iodine (over formaldehyde) . 23
B.3.3 Disadvantages of Lugol’s Iodine . 23
B.4 Ethanol . 24
B.4.1 Advantages of ethanol . 24
B.4.2 Disadvantages of ethanol . 24
Annex C (informative) Corrections of depth from wire angle [1] . 25
Annex D (informative) Example of a field data sheet . 26
Annex E (informative) Ribbon-sampling devices . 27
E.1 Continuous plankton recorder (CPR) . 27
E.2 Longhurst Hardy plankton recorder (LHPR) . 27
Bibliography . 28
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European foreword
This document (EN 17218:2019) has been prepared by Technical Committee CEN/TC 230 “Water
analysis”, the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by November 2019, and conflicting national standards
shall be withdrawn at the latest by November 2019.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organisations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
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Introduction
The Zooplankton community is an important part of the pelagic food web, since it forms the link
between primary producers and higher trophic levels. Changes in phytoplankton biomass and
species/size composition change mesozooplankton community structure and productivity. Such
changes potentially influence fish stock recruitment and sedimentation (i.e. indirectly affecting oxygen
concentration in the bottom water) [1].
Surveys of zooplankton have provided valuable information for the environmental monitoring of
marine and brackish waters, because this group includes species which:
— occur in a wide range of marine and brackish waters over a large geographical area and at the same
time have specific environmental requirements,
— are relatively well known with regard to their geographical distribution and environmental
requirements, and
— have a generally high capacity for dispersal enabling them to respond rapidly to remedial actions,
while sampling requires only a modest expenditure of time and equipment.
A procedure for analysing zooplankton (identification, counting and biomass determination) in marine
and brackish waters is given in EN 17204 [2]. This procedure comprises how to identify and enumerate
zooplankton collected in nets which is utilized to estimate quantitative information on diversity,
abundance and biomass with regard to spatial distribution and long-term temporal trends for a given
body of water.
WARNING — Persons using this document should be familiar with normal laboratory practice. This
document does not purport to address all of the safety problems, if any, associated with its use. It is the
responsibility of the user to establish appropriate safety and health practices.
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1 Scope
This document specifies procedures for sampling of mesozooplankton using nets and continuous
ribbon-sampling devices in marine and brackish waters for the purpose of water quality assessment
and determination of ecological status of ecosystems.
Guidance on sampling procedures and the subsequent steps of preservation and storage are given. The
sampling procedures allow estimates of species occurrence and their abundance (relative or absolute),
including spatial distribution and seasonal and long-term temporal trends, for a given body of water.
The described methods are restricted to the sampling of mesozooplankton that inhabit marine and
brackish waters and exclude the shallow littoral zones which require a different type of sampling (e.g.
zooplankton in salt marshes).
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— IEC Electropedia: available at http://www.electropedia.org/
— ISO Online browsing platform: available at http://www.iso.org/obp
3.1
pelagic zone
free body of water beyond the bottom
3.2
thermocline
layer in a thermally stratified body of water in which the temperature gradient is at a maximum
[SOURCE: ISO 6107-1:2004, 75]
3.3
habitat
area of the environment in which a particular organism lives, including its characteristic assemblages of
plants and animals
Note 1 to entry: It can be either the geographical area over which it extends, or the particular station in which a
specimen is found.
[SOURCE: EN ISO 10870:2012, 2.6, modified – Note 1 to entry has been added]
3.4
biomass concentration
total mass of living organic matter, measured as wet weight, dry weight or ash free dry weight
−3 −3 −3
Note 1 to entry: Unit: g l , g ml , or g m of carbon.
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3.5
plankton
organisms drifting or suspended in water, consisting chiefly of minute plants or animals, but including
larger forms having only weak powers of locomotion
[SOURCE: ISO 6107-5:2004, 41]
3.6
zooplankton
animals present in plankton
[SOURCE: ISO 6107-5:2004, 49]
3.7
mesozooplankton
zooplankton of 0,2 mm to 20 mm size
3.8
sampling site
general area within a body of water from which samples are taken
Note 1 to entry: A site is defined in terms of its location (geographical position, depth) and invariant conditions
(e.g. type of bottom in shallow-water areas) and is delimited on the basis of the accuracy with which these are
given. In cases of doubt when sampling sites have to be re-identified, most weight should be placed on depth and
type of bottom.
[SOURCE: EN ISO 5667-6:2016, 3.10, modified – “or location” is replaced by “within a body of water”
and note 1 to entry has been added]
3.9
sampling station
precise location where samples are collected
Note 1 to entry: A sampling station is defined by its geographical position (latitude, longitude), its depth
(relative to chart datum and normalized to mean low water as given in tide tables) and any other invariant or
physical conditions. The station is delineated using the given level of precision. In cases of doubt, when revisiting
sampling stations, emphasis should be placed on landmarks and water depth.
[SOURCE: EN ISO 16665:2013, 2.2.5]
3.10
trend monitoring
study intended to reveal any changes in variables such as diversity and in the ecological status of a body
of water over time
3.11
preservation
protection from (bio)chemical degradation of organic matter
4 Principle
The sampling strategy determines which information on the current status of the zooplankton
community can be achieved. The selection of sampling sites (numbers and location), sampling depth,
time and frequency of sampling, number of replicates and type of sampling gear is of great importance
for the evaluation of the data collected. As a general guidance EN ISO 5667-1 should be consulted.
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5 Sampling device
5.1 General
The choice of the sampling devices to be used depends on the aims of the investigation. This document
provides some general recommendations and then focuses on standard requirements for net sampling.
Table 1 describes advantages and disadvantages of different common zooplankton sampling devices.
Table 1 — Examples of zooplankton sampling devices
Sampling device Advantages Disadvantages
Simple nets Medium amounts of water can be Can be subject to clogging of mesh.
sampled, can operate easily as
vertical hauls or in restricted
areas.
Multiple nets Large amounts of water can be Difficult to operate in restricted areas.
sampled. Sample can be
separated by different filter sizes
to reduce damage and improve
identification. Allows adjustment
of sampling to physical/biological
conditions (e.g. any
stratification).
High speed samplers Can be towed at higher speeds Difficult to operate in restricted areas.
typically around 9 km/h. Increased risk of damage to delicate
e.g. Gulf VII
organisms.
Continuous recorders Provides spatial information Can Semiquantitative, damage to delicate
operate over very large areas and organisms, e.g. gelatinous
— using ribbons of tape
using vessels of opportunity. mesozooplankton. Limited sampling
e.g. continuous
Used for both phytoplankton and depth.
plankton recorder
zooplankton investigations.
NOTE 1 Several overviews exist on the most widely used zooplankton sampling techniques and their
advantages and drawbacks (e.g. [9, 10, 11]).
NOTE 2 Ribbon-samplers have a fixed method which is largely determined by the internal mechanism and
design of ribbon. Continuous plankton recorder (CPR) devices are designed for us on “vessels of opportunity” so
are also restricted in their range and depth, see Annex E.
5.2 Nets
Polyamide plankton nets with a cod-end and a drain cock of various dimensions and mesh sizes may be
used for sampling (Figure A.1). The purpose of the investigation determines the selection of net types
and its mesh sizes. Examples of commonly-used nets are:
a) Bongo net (Figure A.1);
b) MOCNESS (Multiple Opening and Closing Net with an Environmental Sensing System) [12, 13];
c) WP2 net (Figure A.3);
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d) Multinet (Figures A.4 and A.5);
e) Gulf VII sampler (Figure A.6).
For details, see Annex A.
It is important that nets should have a large filtering surface relative to their opening in order to ensure
that filtering is as efficient as possible. A net with an opening diameter of 30 cm, for example, should
have a length of about one metre as a minimum. A cylindrical net section above the conical part increase
the filtering area compared with a conical plankton net with the same opening diameter and length.
The size of opening itself can determine what is obtained on the mesh. Smaller openings will limit the
capture of faster moving zooplankton and some larger mesozooplankton can evade 1 m ring net. A flow
meter mounted in the net mouth should be used whenever possible.
Closing nets, as opposed to simple open mouthed nets should be used for sampling along transect such
as at discrete depth layers.
NOTE Closing nets remain open until the haul is complete and the mouth or the entrance to the cod-end is
closed. The design and mechanism vary depending on the sampling device being used [13].
Common mesh sizes are e.g. 100 µm in the Baltic Sea or 200 µm up to 500 µm in the North Sea. If early
developmental stages are to be included, in order to provide information on the population dynamics of
zooplankton, nets with a mesh size of up to 50 µm at a maximum are recommended. Mesh sizes above
200 µm miss a large proportion of the smaller zooplankton. Table 2 gives a summary of mesh
requirements for different zooplankton.
Table 2 — Summary of mesh requirements for different zooplankton
Zooplanktonic group Suitable mesh sizes Mesh arrangement
Rotifers, nauplii of crustacea Approx. 50 µm, but > 40 µm Nets with meshes smaller than
(which mostly belong to the 40 µm will readily become
microzooplankton size fraction) clogged and their use should
normally be avoided, although
they may be useful in
oligotrophic waters.
Crustacean plankton only 50 µm (max. 100 µm)
Rotifers and crustaceans, 45 µm for rotifers, 90 µm for 3 nets with 3 different mesh
including predatory most of the crustaceans, and sizes
species
≥ 150 µm for predatory species
Hydromedusae Non-filtering cod-ends should be
used to reduce damage to these
delicate organisms.
All the mesh sizes mentioned in this document should be regarded as for guidance only. Mesh sizes will
also vary somewhat from manufacturer to manufacturer.
It is recommended that, in the case of vertically stratified habitats, the nets are equipped with a closing
mechanism with case weight and a flow counter with backflow stop to allow stratified sampling.
The ribbon-based samplers such as the continuous plankton sampler (Figure A.2) use a band of gauze
rather than a net. In the case of the CPR this is 300 µm mesh. For more on ribbon-based samplers, see
Annex E.
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5.3 Other field equipment
If available, nets should be equipped and deployed with the help of pressure meters so that the actual
vertical position of the net is known.
Field equipment in addition to sampling devices may comprise:
5.3.1 Winch with line-length counter or for coastal areas a line with length markings fitted with a
shackle or similar device to enable the line to be joined to the net.
5.3.2 Flowmeter, either real time or self-logging.
5.3.3 Draining cup with nylon netting, which is capable of being attached to the net either by means
of a tightening strip or tape sewn into the net. The netting of the draining cup should have the same
mesh size as the net. A draining cup with hose and hose clamp can also be utilized.
5.3.4 Weight, e.g. a standard sounding lead weight, in order to minimize wire angles.
5.3.5 Closing device for depth-stratified hauls.
5.3.6 Wire angle blade.
5.3.7 Echosounder or depth finder.
5.3.8 Global Positioning System (GPS).
5.3.9 Sea water connecting tube to flush the net upon retrieval.
5.3.10 Sieves of a mesh size smaller than the net mesh size to concentrate the sample.
5.3.11 Wash bottle with filtrated sea water for rinsing out sieves and draining cups. The sea water
from the sea water hose should be filtrated through a plankton bucket filter, with a small mesh (e.g.
45 µm and always less than the mesh of the sampling devices being used) before filling in the spray
bottle.
5.3.12 Small plastic funnel, may be needed to transfer the sampled material to the sample bottle.
5.3.13 Mixing vessel, e.g. plastic bucket or similar, to combine a number of individual samples into a
single sample in the field. Combining samples may be necessary to reducing analysis times and costs.
5.3.14 Plastic or glass bottles with screw tops for storing samples (e.g. 100 ml, 200 ml or 250 ml,
depending on sampling volume).
5.3.15 Labels or tape to attach to the outside of the sample bottles. Waterproof paper for labels to put
inside the sample bottles.
5.3.16 Marker pen. If ethanol is being used, an alcohol-proof pen or pencil is recommended for both
internal and external marking.
If a volume sampler is being used (with the exception of a Schindler-Patalas trap) filtration equipment
is also required to concentrate the samples. This may take the form of either a plankton net or a large
funnel with draining cup fitted with a netting.
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5.4 Preserving solutions and other chemicals
A number of different preserving solutions for different types of applications are available. The
advantages and disadvantages of each of these solutions are defined in Annex B. Preserving solutions
for field use should be kept in small stoppered bottles and should be accompanied by a pipette or safety
dispenser for transferring the solution to the plankton samples. The bottles should be kept in a plastic
box or container with lid during transportation.
5.4.1 Formaldehyde (CH O), 40 % (v/v).
2
40 % (v/v) formaldehyde is diluted to 4 % formaldehyde (CH O) solution, by mixing 1 part ∽400 ml
2
−1
l formaldehyde solution and 9 parts water. Before diluting the strong formaldehyde it should be
buffered. Disodium tetraborate (borax) (Na B 0 · 10 H 0) or hexamethly tetramine (C H N ) can
2 4 3 2 6 12 4
be used.
In the case of borax buffering, add 2 g of borax to every 98 ml of 40 % formaldehyde. Borax will be in
excess and raise the pH to 8 to 8.2.
In the case of hexamethyl tetramine buffering, dilute the formaldehyde with demineralized water to
20 % (v/v) to avoid precipitation, and then add 100 g of hexamethyl tetramine and 40 g to 80 g sucrose
per litre of 20 % formaldehyde [11].
WARNING — Formaldehyde may trigger allergies or cancers and should therefore be handled with
care.
5.4.2 Ethanol, (C H OH), 96 % (v/v) or 99 % (v/v).
2 5
5.4.3 Lugol’s Iodine.
Acidified Lugol’s Iodine: Dissolve 100 g KI (potassium iodide) in 1 l of distilled or demineralized water;
then add 50 g iodine (crystalline), shake until it is dissolved and add 100 ml of glacial (anhydrous)
acetic acid. As this solution is close to saturation, any precipitate should be removed by decanting the
solution before use.
NOTE Acidic Lugol’s can dissolve calcareous skeletons of zooplankton.
5.4.4 Saccharose, 40 g to 80 g per litre formaldehyde.
5.4.5 Anti-fungal agents, such as Steedman’s Observation fluid. This fluid combines 0,5 % propylene
phenoxetol and 5 % propane-1-2-diol in distilled water. Which is then added to samples originally
preserved with formaldehyde [14].
5.4.6 Mastail and Battaglia solution. Prepare separate solutions by dissolving 8 g
buthylhydroxyanisol (BHA, C H O) in 500 ml propane-1-2-diol (C H O) and 20 g
22 32 4 3 8 2
ethylenediaminetetraacetic acid (EDTA, C H N O Na · 2 H O) in 500 ml demineralized water. Add
10 14 2 8 2 2
−1
both solutions to 2 L of ∽400 ml l of formaldehyde solution while stirring and buffer to pH 8 with
sodium glycerophosphate (C H Na O P · H O). After buffering add 2 g ascorbic acid (C H O ) and
3 7 2 6 2 6 8 6
demineralized water up to 5 L. Samples are preserved by adding 6 ml of the stock solution per 100 ml of
sample in sea water.
NOTE This solution i
...
SLOVENSKI STANDARD
oSIST prEN 17218:2018
01-marec-2018
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Water quality - Guidance for the sampling of mesozooplankton from marine and brackish
waters using mesh
Wasserbeschaffenheit - Anleitung für die Probenahme von Mesozooplankton aus
marinen und Übergangsgewässern mittels Netzen
Qualité de l'eau - Document d'orientation pour l'échantillonnage du mésozooplancton
dans les eaux de mer ou saumâtres à l'aide de filets
Ta slovenski standard je istoveten z: prEN 17218
ICS:
13.060.10 Voda iz naravnih virov Water of natural resources
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
oSIST prEN 17218:2018 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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oSIST prEN 17218:2018
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oSIST prEN 17218:2018
DRAFT
EUROPEAN STANDARD
prEN 17218
NORME EUROPÉENNE
EUROPÄISCHE NORM
January 2018
ICS 13.060.70
English Version
Water quality - Guidance on sampling of mesozooplankton
from marine and brackish water using mesh
Qualité de l'eau - Document d'orientation pour Wasserbeschaffenheit - Anleitung zur Probenahme von
l'échantillonnage du mésozooplancton dans les eaux de Mesozooplankton aus marinen und
mer ou saumâtres à l'aide de filets Übergangsgewässern mittels Netzen
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 230.
If this draft becomes a European Standard, CEN members are bound to comply with the CEN/CENELEC Internal Regulations
which stipulate the conditions for giving this European Standard the status of a national standard without any alteration.
This draft European Standard was established by CEN in three official versions (English, French, German). A version in any other
language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC
Management Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and United Kingdom.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.
Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2018 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 17218:2018 E
worldwide for CEN national Members.
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oSIST prEN 17218:2018
prEN 17218:2018 (E)
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 7
5 Sampling device . 8
5.1 General . 8
5.2 Nets . 9
5.3 Other field equipment . 10
5.4 Preserving solutions and other chemicals . 11
6 Prearrangements of sampling . 12
6.1 Documentation of strategies and methods . 12
6.2 Preparation of sampling equipment . 12
6.3 Safety instructions . 13
7 Sampling procedure . 13
7.1 Investigation programme . 13
7.2 Number and location of sampling sites . 13
7.2.1 General . 13
7.3 Diurnal sampling period . 14
7.3.1 General . 14
7.3.2 Sample size . 14
7.3.3 Geographical localization of sampling sites . 15
7.4 Operating the sampling device . 15
7.4.1 Vertical Net hauls . 15
7.4.2 Horizontal Tows/Hauls . 16
7.4.3 Sampling jellyfish . 17
7.4.4 Filling and labelling of sample bottles . 17
7.4.5 Preservation and storage of samples . 17
7.5 Field data recording . 18
8 Quality assurance . 18
Annex A (informative) Examples of sampling devices . 20
A.1 Bongo Net . 20
A.2 Continuous Plankton Recorder . 20
A.3 WP2 Net . 21
A.4 Multi nets . 21
A.5 Gulf VII Sampler . 22
Annex B (informative) Preservation . 24
B.1 Preservation. 24
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B.2 Formaldehyde (formalin) . 24
B.2.1 General . 24
B.2.2 Advantages of formaldehyde . 25
B.2.3 Disadvantages of formaldehyde . 25
B.3 Lugol’s Iodine . 25
B.3.1 General . 25
B.3.2 Advantages of Lugol’s Iodine (over formaldehyde) . 25
B.3.3 Disadvantages of Lugol’s Iodine . 25
B.4 Ethanol . 26
B.4.1 Advantages of ethanol . 26
B.4.2 Disadvantages of ethanol . 26
Annex C (informative) Corrections of depth from wire angle [4]. 27
Annex D (informative) Example of a field data sheet . 29
Annex E (informative) Ribbon-sampling devices . 30
E.1 Continuous Plankton Recorder (CPR) . 30
E.2 Longhurst Hardy Plankton Recorder (LHPR). 30
Bibliography . 31
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European foreword
This document (prEN 17218:2018) has been prepared by Technical Committee CEN/TC 230 “Water
analysis”, the secretariat of which is held by DIN.
This document is currently submitted to the CEN Enquiry.
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Introduction
Zooplankton abundance and community structure constitute an important part of the pelagic food web,
since it forms the link between primary producers and higher trophic levels. Changes in phytoplankton
biomass and species/size composition change mesozooplankton community structure and productivity.
Such changes potentially influence fish stock recruitment and sedimentation (i.e. indirectly affecting
oxygen concentration in the bottom water) [4].
Surveys of zooplankton have provided valuable information for the environmental monitoring of
marine and brackish waters, because this group includes species which:
— occur in a wide range of marine and brackish waters over a large geographical area and at the same
time have specific environmental requirements;
— are well known with regard to their geographical distribution and environmental requirements;
— have a generally high capacity for dispersal enabling them to respond rapidly to remedial actions;
while
— sampling requires only a modest expenditure of time and equipment.
A procedure for analysing zooplankton (identification, counting and biomass determination) in marine
and brackish waters is given in prEN 17204. This procedure comprises how to identify and enumerate
zooplankton collected in nets which is utilized to estimate quantitative information on diversity,
abundance and biomass with regard to spatial distribution and long-term temporal trends for a given
body of water.
WARNING — Persons using this document should be familiar with normal laboratory practice.
This document does not purport to address all of the safety problems, if any, associated with its
use. It is the responsibility of the user to establish appropriate safety and health practices.
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1 Scope
This document specifies procedures for sampling of mesozooplankton using nets and ribbon-sampling
devices in marine and brackish waters for the purpose of water quality assessment and determination
of ecological status of ecosystems.
Guidance on sampling procedures and the subsequent steps of preservation and storage are given. The
sampling procedures allow estimates of species occurrence and their abundance (relative or absolute),
including spatial distribution and seasonal and long-term temporal trends, for a given body of water.
The described methods are restricted to the sampling of mesozooplankton that inhabit marine and
brackish waters and exclude the shallow littoral zones which require a different type of sampling (e.g.
zooplankton in salt marshes).
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN ISO 5667-3, Water quality — Sampling — Part 3: Preservation and handling of water samples (ISO
5667-3)
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— IEC Electropedia: available at http://www.electropedia.org/
— ISO Online browsing platform: available at http://www.iso.org/obp
3.1
pelagic zone
free body of water beyond the bottom
3.2
thermocline
layer in a thermally stratified body of water in which the temperature gradient is at a maximum
3.3
habitat
area of the environment in which a particular organism lives, including its characteristic assemblages of
plants and animals
Note 1 to entry: It can be either the geographical area over which it extends, or the particular station in which a
specimen is found.
[SOURCE: EN ISO 10870:2012, 2.6, modified – Note 1 to entry has been added]
3.4
biomass concentration
−3 −3 −3
total mass (unit: g l , g ml , or g carbon m ) of living organic matter within a taxon. Measured as
wet weight, dry weight or ash free dry weight
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3.5
plankton
organisms drifting or suspended in water, consisting chiefly of minute plants or animals, but including
larger forms having only weak powers of locomotion
[SOURCE: ISO 6107-5:2004, definition 41]
3.6
zooplankton
animals present in plankton
[SOURCE: ISO 6107-5:2004, definition 49]
3.7
mesozooplankton
zooplankton of 0,2 mm to 20 mm size
3.8
sampling site
general area within a body of water from which samples are taken
Note 1 to entry: A station is defined in terms of its location (geographical position, depth) and invariant
conditions (e.g. type of bottom in shallow-water areas) and is delimited on the basis of the accuracy with which
these are given. In cases of doubt when sampling stations have to be re-identified, most weight should be placed
on depth and type of bottom.
3.9
sampling station
precise location where samples are collected
Note 1 to entry: A sampling station is defined by its geographical position (latitude, longitude), its depth
(relative to chart datum and normalized to mean low water as given in tide tables) and any other invariant or
physical conditions. The station is delineated using the given level of precision possible when revisiting sampling
stations. Emphasis should be placed on landmarks and water depth.
[SOURCE: EN ISO 16665:2013, definition 2.2.5]
3.10
trend monitoring
study intended to reveal any changes in diversity and in the ecological status of a body of water over
time
3.11
preservation
protection from (bio) chemical degradation of organic matter
4 Principle
The sampling strategy determines which information on the current status of the zooplankton
community can be achieved. The selection of sampling sites (numbers and location), sampling depth,
time and frequency of sampling, number of replicates and type of sampling gear is of great importance
for the evaluation of the data collected. As a general guidance EN ISO 5667-1 should be consulted.
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5 Sampling device
5.1 General
The choice of the sampling devices to be used depends on the aims of the investigation. This document
provides some general recommendations and then focuses on standard requirements for net sampling.
Table 1 describes advantages and disadvantages of different common zooplankton sampling devices.
Table 1 — Examples of zooplankton sampling devices
Sampling device Advantages Disadvantages
Smaller amounts of water may be
Simple operation. Can operate in
Volume samplers sampled. Required secondary
shallow waters
processing to concentrate sample
Can be damaging to gelatinous
Medium amounts of water can be
mesozooplankton particularly if using
Pumps sampled. Can operate in all
a narrow bore hose. Can be restricted
shallow waters
to shallow depths
Medium amounts of water can be
Can be more subject to clogging of
sampled, can operate easily as
Simple nets
mesh
vertically hauls or in restricted
areas
Large amounts of water can be
sampled. Sample can be
separated by different filter sizes
Multiple nets to reduce damage and improve Difficult to operate in restricted areas
identification. Allows adjustment
of sampling to physical/biological
conditions (e.g. any stratification)
Difficult to operate in restricted
High speed samplers
Can be towed at higher speeds
areas. Increased risk of damage to
typically 5 kn
e.g. Gulf VII
delicate organisms
Provides spatial information Can
Continuous recorders
Semiquantitative, damage to delicate
operate over very large areas and
organisms, e.g. gelatinous
— using ribbons of
using vessels of opportunity.
mesozooplankton. Limited sampling
tape e.g. continuous
Used for both phyto- and
depth
plankton recorder
zooplankton investigations
NOTE 1 Several overviews exist on the most widely used zooplankton sampling techniques and their
advantages and drawbacks (e.g. [5, 6, 7]).
NOTE 2 Ribbon-samplers have a fixed method which is largely determined by the internal mechanism and
design of ribbon. Continuous plankton recorder (CPR) devices are designed for us on “vessels of opportunity” so
are also restricted in their range and depth, see Annex E.
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5.2 Nets
Polyamide plankton nets with a cod-end and a drain cock of various dimensions and mesh sizes may be
used for sampling (Figure A.1). The purpose of the investigation determines the selection of net types
and its mesh sizes.
Examples of commonly-used nets are:
a) Bongo-net (Figure A.1);
b) MOCNESS (Multiple Opening and Closing Net with an Environmental Sensing System) [8, 9];
c) WP2 net (Figure A.3);
d) Multinet (Figure A.4);
e) Gulf VII sampler (Figure A.7).
For details, see Annex A.
It is important that nets should have a large filtering surface relative to their opening in order to ensure
that filtering is as efficient as possible. A net with an opening diameter of 30 cm, for example, should
have a length of about one metre as a minimum. A cylindrical net section above the conical part increase
the filtering area compared with a conical plankton net with the same opening diameter and length.
The size of opening itself can determine what is obtained on the mesh. Smaller openings will limit the
capture of faster moving zooplankton and some larger mesozooplankton can evade 1 m ring net. A flow
meter mounted in the net mouth should be used whenever possible.
Closing nets, as opposed to simple open mouthed nets should be used for sampling along transect such
as at discrete depth layers.
NOTE Closing nets remain open until the haul is complete and the mouth or the entrance to the cod-end is
closed. The design and mechanism varies depending on the sampling device being used [9].
Common mesh sizes are e.g. 100 µm in the Baltic Sea or 200 µm up to 500 µm in the North Sea. If early
developmental stages are to be included, in order to provide information on the population dynamics of
zooplankton, nets with a mesh size of up to 50 µm at a maximum are recommended. Mesh sizes above
200 µm miss a large proportion of the smaller zooplankton. Table 2 gives a summary of mesh
requirements for different zooplankton.
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Table 2 — Summary of mesh requirements for different zooplankton
Zooplanktonic group Suitable mesh sizes Mesh arrangement
Nets with meshes smaller than
40 µm will readily become
Rotifers, nauplii of crustacea
clogged and their use should
(which mostly belong to the Approx. 50 µm, but > 40 µm
normally be avoided, although
microzooplankton size fraction)
they may be useful in oligotrophic
waters.
Crustacean plankton only 50 µm (max. 100 µm)
45 µm for rotifers, 90 µm for most
Rotifers and crustaceans,
of the crustaceans, and
3 nets with 3 different mesh sizes
including predatory species
≥ 150 µm for predatory species
Non-filtering cod-ends should be
Hydromedusae used to reduce damage to these
delicate organisms.
All the mesh sizes mentioned in this document should be regarded as for guidance only. Mesh sizes will
also vary somewhat from manufacturer to manufacturer.
It is recommended that, in the case of vertically stratified habitats, the nets are equipped with a closing
mechanism with case weight and a flow counter with backflow stop to allow stratified sampling.
The ribbon-based samplers such as the Continuous plankton sampler (Figure A.7) use a band of gauze
rather than a net. In the case of the CPR this is 300 micron mesh. For more on ribbon-based samplers,
see Annex E.
5.3 Other field equipment
If available, nets should be equipped and deployed with the help of pressure meters so that the actual
vertical position of the net is known.
Field equipment in addition to sampling devices may comprise:
5.3.1 Winch with line-length counter or for coastal areas a line with length markings fitted with a
shackle or similar device to enable the line to be joined to the net.
5.3.2 Flowmeter – either real time or self-logging.
5.3.3 Draining cup with nylon netting, which is capable of being attached to the net either by means
of a tightening strip or tape sewn into the net. The netting of the draining cup should have the same
mesh size as the net. A draining cup with hose and hose clamp can also be utilized.
5.3.4 Weight, e.g. a standard sounding lead weight, in order to minimize wire angles.
5.3.5 Closing device for depth-stratified hauls.
5.3.6 Wire angle blade.
5.3.7 Echosounder or depth finder.
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5.3.8 Global Positioning System (GPS).
5.3.9 Sea water connecting tube to flush the net upon retrieval.
5.3.10 Sieves of a mesh size smaller than the net mesh size to concentrate the sample.
5.3.11 Wash bottle with filtrated sea water for rinsing out sieves and draining cups. The sea water
from the sea water hose should be filtrated through a plankton bucket filter, with a small mesh e.g.
45 µm and always less than the mesh of the sampling devices being used) before filling in the spray
bottle.
5.3.12 Small plastic funnel, may be needed to transfer the sampled material to the sample bottle.
5.3.13 Mixing vessel, e.g. plastic bucket or similar, to combine a number of individual samples into a
single sample in the field. Conducting mixed samples may be necessary to reducing analysis times and
costs.
5.3.14 Plastic or glass bottles with screw tops for storing samples (e.g. 100 ml, 200 ml or 250 ml,
depending on sampling volume).
5.3.15 Labels or tape to attach to the outside of the sample bottles. Waterproof paper for labels to put
inside the sample bottles.
5.3.16 Marker pen. If ethanol is being used, an alcohol-proof pen or pencil is recommended for both
internal and external marking.
If a volume sampler is being used (with the exception of a Schindler-Patalas trap) filtration equipment
is also required to concentrate the samples. This may take the form of either a plankton net or a large
funnel with draining cup fitted with a netting.
5.4 Preserving solutions and other chemicals
A number of different preserving solutions for different types of applications are available. The
advantages and disadvantages of each of these solutions are defined in Annex B. Preserving solutions
for field use should be kept in small stoppered bottles and should be accompanied by a pipette or safety
dispenser for transferring the solution to the plankton samples. The bottles should be kept in a plastic
box or container with lid during transportation.
5.4.1 Formaldehyde (HCHO), 40 % by volume.
This is neutralized, e.g. with hexamethyl tetramine (C H N ) or buffered to pH 8 to 8,2 with disodium
6 12 4
tetraborate (borax) (Na B 0 · 10 H 0). Before diluting the strong formaldehyde, add 2 g of borax to
2 4 3 2
every 98 ml of 40 % formaldehyde. Borax will be in excess and raise the pH to 8 to 8,2.
Then dilute the formaldehyde with water to 20 % (v/v) to avoid precipitation, and then add 100 g of
hexamethyl tetramine and 40 g to 80 g sucrose per litre of 20 % formaldehyde [7].
WARNING — Formaldehyde may trigger allergies or cancers, and should therefore be handled with
care.
5.4.2 Ethanol, 96 % or 99 % C H OH.
2 5
5.4.3 Lugol’s Iodine.
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Acidified Lugol’s Iodine: Dissolve 100 g KI (potassium iodide) in 1 l of distilled or demineralized water;
then add 50 g iodine (crystalline), shake until it is dissolved and add 100 ml of glacial acetic acid. As this
solution is close to saturation, any precipitate should be removed by decanting the solution before use.
NOTE Acidic Lugol’s can dissolve calcareous skeletons of zooplankton.
5.4.4 Saccharose, 40 g to 80 g per litre formaldehyde.
5.4.5 Anti-fungal agents, such as Steedman’s Observation fluid. This fluid uses propylene phenoxetol
and propylene glycol in combination to samples originally preserved with formaldehyde. [10]
6 Prearrangements of sampling
6.1 Documentation of strategies and methods
The following documentation should be available before the start of field work:
a) description of objectives and strategy;
b) description of methods;
c) safety instructions;
d) personnel plan;
e) overview of equipment and instruments;
f) registration forms;
g) procedures for the maintenance of records and samples;
h) quality assurance requirements according to EN 14996 and EN ISO IEC 17025;
i) chemical safety data sheets.
6.2 Preparation of sampling equipment
Nets should be clean to avoid contamination of samples between sites,
...
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