SIST EN 15972:2012

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Kakovost vode - Navodilo za kvantitativne in kvalitativne preiskave morskega fitoplanktonaWasserbeschaffenheit - Anleitung für die quantitative und qualitative Untersuchung von marinem PhytoplanktonQualité de l'eau - Guide pour l'étude quantitative et qualitative du phytoplancton marinWater quality - Guidance on quantitative and qualitative investigations of marine phytoplankton13.060.70Preiskava bioloških lastnosti vodeExamination of biological properties of water13.060.10Voda iz naravnih virovWater of natural resourcesICS:Ta slovenski standard je istoveten z:EN 15972:2011SIST EN 15972:2012en,fr,de01-februar-2012SIST EN 15972:2012SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 15972
September 2011 ICS 13.060.70 English Version
Water quality - Guidance on quantitative and qualitative investigations of marine phytoplankton
Qualité de l'eau - Guide pour l'étude quantitative et qualitative du phytoplancton marin
Wasserbeschaffenheit - Anleitung für die quantitative und qualitative Untersuchung von marinem Phytoplankton This European Standard was approved by CEN on 29 July 2011.
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.
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Sampling and sample depth . 19A.1General . 19A.2Sampling depth . 19A.3Integrated samples . 19A.4Combined samples . 20Annex B (informative)
Water samplers . 21Annex C (informative)
Plankton net samples . 22Annex D (informative)
Preservatives . 23SIST EN 15972:2012

Counting chambers for quantification without concentration of the phytoplankton cells . 25E.1Sedgwick – Rafter . 25E.2Palmer – Maloney . 25E.3Haemacytometre . 25Annex F (informative)
Utermöhl method . 26Bibliography . 27 SIST EN 15972:2012

phytoplankton NOTE The phytoplankton biomass can be expressed in terms of chlorophyll content, the carbon content of the algae (algal carbon) or the volume of the phytoplankton (biovolume). 3.4 qualitative sample phytoplankton sample collected by plankton net, i.e. a concentrated sample where the size of the phytoplankton is mainly larger than the mesh diameter NOTE Allows for recordings of taxon/taxa occurring in low concentrations. 3.5 quantitative sample phytoplankton sample where a part of a body of water is enclosed so that the density of each taxon and their relative abundance per unit volume may be recorded 3.6 receiving water body water body that receives an input of material, of either natural or anthropogenic origin NOTE The term often appears in the context of anthropogenic input, for example, effluent from municipal waste water outlets or industrial processed water. [EN ISO 16665:2005, 2.4] SIST EN 15972:2012

When using this means of sampling, the samples can become contaminated by micro-algae associated with hard substrates such as fouling on the quay/pier, macroalgae etc. The samples shall therefore be taken upstream and as far from the edge of the quay/pier as possible. For baseline surveys in receiving waters from industry, municipal discharges and aquaculture activities, the samples shall, as far as possible, be taken upstream of the plant(s). Sampling at a single station is usually sufficient. However, in particular or acute situations, sampling shall be carried out according to continual assessment of the most beneficial station positioning. 4.4 Sampling depth In the case of a monitoring network operated in an international framework, sampling strategy shall be elaborated by the expert group of the member states. Samples for quantitative analysis may be collected either individually from fixed depths, as combined samples where samples from different depths are mixed together to form a single sample or as integrated samples, where sampling encompasses the entire water column. Combined samples may be used in cases when an overview of the numerical dominating species is required and where it is not necessary to link the occurrence of phytoplankton to hydrographical conditions or chemical parameters. In marine areas, where large fluctuations in the physical parameters within the water column occur, such as fjords or brackish waters, the samples should be taken both above and below the thermocline, because they can often form a boundary between different phytoplankton stocks. In marine areas where different water bodies meet, hydrographic measurements shall be carried out to identify stratified water layers of various origins down the water column, in order to ensure that the different water masses are adequately sampled. For baseline surveys or trend monitoring, where the phytoplankton occurrences are related to chemical parameters such as nutrient levels, samples usually shall be taken from fixed depths. Standard sampling depths within the upper 100 m of open sea (recommended by ICES1) are 0 m, 5 m, 10 m, 20 m, 30 m, 50 m, 75 m and 100 m. This should be the case in areas where no vertical stratification is found. In fjord areas, additional sampling shall be carried out at 2 m. The number of sampling depths may be increased if there is information from, for example, hydrographic measurements on increased frequency of phytoplankton occurrence other than the recommended depth. If in situ fluorescence is measured, an additional sample may be collected from the depth at which the fluorescence maximum occurs. See A.2.2 for a description of fluorescence maximum. The sampling depth from which samples are to be processed should be decided on an individual basis, based on the aim of the investigation. As a minimum, at least one water sample from every sampling date shall be processed. Samples from the same depths shall be processed throughout the entire duration of the programme, to ensure comparable data. Additional samples can be processed if necessary to get information about the phytoplankton composition at the thermocline or other interesting levels in the water column. Integrated samples are often used in connection with specialised surveys and monitoring of particular taxon/taxa. For example, for monitoring of toxic or harmful phytoplankton in connection with fish or shellfish production, integrated samples shall be taken within the depth intervals at which the fish or shellfish are located if vertical stratification occur in the height of water concerned by the aquaculture livestock. The net samples are used to complete the total species spectrum and for the determination of difficult taxa. The precise depth, or depth interval, at which the haul shall be carried out, shall be decided based on the aims of the sampling programme and/or the hydrographic conditions at the sampling location.
1 International Council for the Exploration of the Sea SIST EN 15972:2012

6 Survey procedures 6.1 Description of procedures Before the start of field work, descriptions of methodology, safety instructions, personnel plan, overview of equipment and instruments, recording form, instructions for recording, procedures for securing records and samples as well as information on quality assurance requirements should be compiled.
6.2 Preparation The following shall be carried out during equipment preparation:  check that the plankton net is clean and in good condition, without holes or tears;  check that the attachment of the rope/wire to the plankton net is sufficiently secure;  rope shall be marked with metre marks in advance. The rope used shall not stretch when wet;  check that the water sampler’s closing mechanism is functioning well and that any seals are in order. 6.3 Defining the position of sampling stations The position of sampling stations shall be defined unambiguously, such that they can easily be relocated by other investigators. Positions shall be defined using geographic co-ordinates and in accordance with a reference system (e.g. European Datum: ED-50, World Geodetic System: WGS-84), and also in relation to the UTM-system if appropriate. Positions shall be defined according to relevant guidelines. When sampling around an aquaculture installation, the concession number shall be provided. When sampling from a boat the sampling strategy should specify if drift during sampling has to be prevented or not. 6.4 Operating the sampling device
6.4.1 Water samplers and devices for collecting integrated samples When deploying a water sampler, the closing mechanism shall be checked to ensure it is properly open to allow an unhindered water throughflow. The water sampler shall be lowered vertically to the required depth at an even, moderate speed and then closed. Hose samplers and other samplers for integrated sampling shall be lowered slowly to the required depth such that the water column from which the sample shall be taken is disturbed as little as possible by the sampling device. 6.4.2 Plankton net hauls The plankton net shall be lowered to the required depth at which the sampling should either be carried out at or start from, and hauled vertically, at an angle or towed horizontally. For vertical hauls and those at an angle, SIST EN 15972:2012

Prior to sampling at a new station, the plankton net shall be rinsed well, preferably with fresh water. If fresh water is not available, the equipment shall be rinsed with water from the new sampling location before it is deployed. The collecting cup shall be removed before rinsing or alternatively, if the net has a fixed collecting cup with a tap, this shall be opened to ensure a good throughflow of water. 6.5 Sample fixation Fixing of phytoplankton shall be carried out immediately after the sample is transferred to the sample bottle in order to avoid loss of phytoplankton cells. For quantitative analysis, 0,2 ml to 1,0 ml of Lugol’s solution is added per 100 ml of water sample [13]. During phytoplankton blooming time, during which there is a very high concentration of phytoplankton, the quantity of fixative may be increased up to 2 ml per 100 ml sample. The sample should be golden brown in colour after fixation. For qualitative analysis, plankton net samples should be fixed using a 20 % formaldehyde solution buffered with hexamethylentetramin (C6H12N4) in a 3:1 sample to formaldehyde solution proportion. The quantity of fixative can be reduced considerably if there is only sparse material in the sample. 6.6 Samples for analysis of living material If living phytoplankton shall be identified and quantified, the samples shall be kept dark and at a constant temperature equivalent to that at the sampling depth. If the water temperature at the sampling location is higher than 10 °C, the sample shall gradually be cooled down to 10 °C. NOTE Warming of the sample, for example by exposure to sunlight, can result in destruction of phytoplankton cells. Samples for analysis of living phytoplankton shall not be stored longer than 36 h. If both living and preserved material is to be examined, a double set of samples shall be collected; one for analysis of living material and one sample for preserved material. 6.7 Sample labelling and recording of additional information Sample bottles shall be labelled. The label should contain the following information:  name of site or sampling locality;  UTM coordinates (or coordinates given in another specified geographic reference system if appropriate); SIST EN 15972:2012

7.1.2 Specialised surveys - monitoring of selected taxa For monitoring of harmful and/or toxic phytoplankton, selected taxa shall be identified according to a pre-defined list. The taxa listed will depend on the situation and aim of the monitoring investigation. For issuing guidelines on shellfish consumption, the emphasis shall be placed on species toxic to humans, as highlighted by the relevant authorities. For monitoring around fish farms, the emphasis shall be placed on potentially ichthyotoxic species. 7.2 Methods for quantification 7.2.1 Methods and detection limits Various light microscopy methods exist for quantification of plankton algae. Phase contrast is needed to quantify many almost transparent cells. For surveys requiring a description of species composition, the sample material should usually be concentrated because many species naturally occur in low concentrations. The sample material may be concentrated by use of different types of filtration methods or by sedimentation in a sedimentation chamber, see EN 15204 for more detailed descriptions of quantification methods. Taxon/taxa that occur in relatively high concentrations may be quantified directly from the sample, without prior concentration. NOTE Such methods have higher detection limits because very small sample volumes are analysed and are therefore suitable for quantification of taxon/taxa that occur in high concentrations. Before taking sub-samples, the samples shall be adapted to room temperature. The samples shall be thoroughly mixed by gently inverting and righting the bottle for 1 min to 3 min. Table 1 gives a selection of methods with their detection limits. Table 1 — Selected quantification met
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