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

Status
Published
Public Enquiry End Date
01-Apr-2018
Publication Date
04-Jun-2019
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
14-May-2019
Due Date
19-Jul-2019
Completion Date
05-Jun-2019

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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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SIST EN 17218:2019
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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EN 17218:2019 (E)
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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EN 17218:2019 (E)
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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EN 17218:2019 (E)
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).

40 % (v/v) formaldehyde is diluted to 4 % formaldehyde (CH O) solution, by mixing 1 part ∽400 ml

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

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
...

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