Water quality - Determination of acute toxicity of marine or estuarine sediment to amphipods (ISO 16712:2005)

This International Standard specifies a method for the determination of acute toxicity to amphipods exposed over a period of 10 d to a) samples of contaminated marine or estuarine sediment, b) chemical, industrial or municipal sludge, or other solid wastes that may combine with marine or estuarine sediments, or c) chemicals or preparations spiked into clean sediment.

Wasserbeschaffenheit - Bestimmung der akuten Toxizität mariner Sedimente oder von Sedimenten aus Flussmündungsgebieten gegenüber Amphipoden (ISO 16712:2005)

Diese Internationale Norm legt ein Verfahren zur Bestimmung der akuten Toxizität gegenüber Amphipoden fest, die über einen Zeitraum von 10 Tagen
a)   Proben von belasteten marinen Sedimenten oder Sedimenten aus Flussmündungsgebieten,
b)   chemischem, industriellem oder kommunalem Klärschlamm oder anderen festen Abfällen, die in marinen Sedimenten oder Sedimenten aus Flussmündungsgebieten vorhanden sein können, oder
c)   Chemikalien oder Ansätzen, mit denen unbelastete Sedimenten aufgestockt sind,
ausgesetzt wurden.

Qualité de l'eau - Détermination de la toxicité aiguë des sédiments marins et estuariens vis-a-vis des amphipodes (ISO 16712:2005)

L'ISO 16712:2005 spécifie une méthode de détermination de la toxicité aiguë vis-à-vis des amphipodes exposés pendant une période de 10 jours à des échantillons de sédiment marin ou estuarien pollué, à des boues chimiques, industrielles ou urbaines, ou à d'autres déchets solides susceptibles de s'associer aux sédiments marins ou estuariens, ou à du sédiment propre dopé avec des produits chimiques ou des préparations.

Kakovost vode - Določanje akutne toksičnosti morskih sedimentov ali sedimentov iz rečnih ustij na rake (Amphipoda) (ISO 16712:2005)

General Information

Status
Published
Publication Date
31-Jan-2007
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
01-Feb-2007
Due Date
01-Feb-2007
Completion Date
01-Feb-2007

Buy Standard

Standard
EN ISO 16712:2007
English language
24 pages
sale 10% off
Preview
sale 10% off
Preview
e-Library read for
1 day

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Water quality - Determination of acute toxicity of marine or estuarine sediment to amphipods (ISO 16712:2005)Qualité de l'eau - Détermination de la toxicité aiguë des sédiments marins et estuariens vis-a-vis des amphipodes (ISO 16712:2005)Wasserbeschaffenheit - Bestimmung der akuten Toxizität mariner Sedimente oder von Sedimenten aus Flussmündungsgebieten gegenüber Amphipoden (ISO 16712:2005)13.080.30Biološke lastnosti talBiological properties of soils13.060.10Voda iz naravnih virovWater of natural resourcesICS:SIST EN ISO 16712:2007en,fr,deTa slovenski standard je istoveten z:EN ISO 16712:200601-februar-2007SIST EN ISO 16712:2007SLOVENSKI
STANDARD







EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN ISO 16712October 2006ICS 13.080.30 English VersionWater quality - Determination of acute toxicity of marine orestuarine sediment to amphipods (ISO 16712:2005)Qualité de l'eau - Détermination de la toxicité aiguë dessédiments marins et estuariens vis-à-vis des amphipodes(ISO 16712:2005)Wasserbeschaffenheit - Bestimmung der akuten Toxizitätmariner Sedimente oder von Sedimenten ausFlussmündungsgebieten gegenüber Amphipoden (ISO16712:2005)This European Standard was approved by CEN on 11 September 2006.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the Central Secretariat or to any CEN member.This European Standard exists in three official versions (English, French, German). A version in any other language made by translationunder the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania,Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2006 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 16712:2006: E



EN ISO 16712:2006 (E)
2
Foreword
The text of ISO 16712:2005 has been prepared by Technical Committee ISO/TC 147 "Water quality” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 16712:2006 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 April 2007, and conflicting national standards shall be withdrawn at the latest by April 2007.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
Endorsement notice
The text of ISO 16712:2005 has been approved by CEN as EN ISO 16712:2006 without any modifications.



INTERNATIONALSTANDARDISO16712First edition2005-02-15Reference numberISO16712:2005(E)© ISO2005Water quality— Determination of acute toxicity of marine or estuarine sediment to amphipodsQualité de l'eau— Détermination de la toxicité aiguë des sédiments marins et estuariens vis-à-vis des amphipodes



ISO16712:2005(E)ii© ISO2005–All rights reservedPDF disclaimerThis PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but shallnot be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. Indownloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariataccepts no liability in this area.Adobe is a trademark of Adobe Systems Incorporated.Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creationparameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In theunlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.©ISO2005All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below orISO's member body in the country of the requester.ISO copyright officeCase postale 56  CH-1211 Geneva 20Tel.+ 41 22 749 01 11Fax+ 41 22 749 09 47E-mailcopyright@iso.orgWebwww.iso.orgPublished in Switzerland



ISO16712:2005(E)© ISO2005–All rights reservediiiContents Page1Scope . 12Principle . 13Test environment . 13.1Facilities . 13.2Lighting . 14Reagents and materials . 14.1Test organism . 14.2Overlying water . 35Apparatus . 46Treatment and preparation of samples . 56.1General . 56.2Control or reference sediment . 56.3Test sediment . 56.4Preparation of sediment samples . 57Test procedures . 67.1Preparing the exposure vessels . 67.2Introducing the organisms . 67.3Test conditions . 77.4Test observations and measurements . 77.5Expression of results . 87.6Reburrowing capability . 87.7Validity of the test . 88Analysis and interpretation of results . 88.1Data analysis . 88.2Non-contaminant factors . 99Reference toxicant . 910Precision . 911Test report . 10AnnexA(informative)Reconstitued salt water . 11AnnexB(normative)Maximum length of amphipod species, optimal salinity and temperatureranges . 12AnnexC(informative)Precision data . 13Bibliography . 14



ISO16712:2005(E)iv© ISO2005–All rights reservedForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies(ISO member bodies). The work of preparing International Standards is normally carried out through ISOtechnical committees. Each member body interested in a subject for which a technical committee has beenestablished has the right to be represented on that committee. International organizations, governmental andnon-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the InternationalElectrotechnical Commission (IEC) on all matters of electrotechnical standardization.International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part2.The main task of technical committees is to prepare International Standards. Draft International Standardsadopted by the technical committees are circulated to the member bodies for voting. Publication as anInternational Standard requires approval by at least 75% of the member bodies casting a vote.Attention is drawn to the possibility that some of the elements of this document may be the subject of patentrights. ISO shall not be held responsible for identifying any or all such patent rights.ISO16712 was prepared by Technical Committee ISO/TC147, Water quality, Subcommittee SC5, Biologicalmethods.



ISO16712:2005(E)© ISO2005–All rights reservedvIntroductionThis International Standard outlines procedures for conducting acute tests for sediment toxicity, using one ormore amphipod species that are found primarily below the sediment surface in coastal marine and estuarinewaters. The biological endpoint for the test is percent mortality at day 10.



.vi



INTERNATIONAL STANDARDISO16712:2005(E)© ISO2005–All rights reserved1Water quality— Determination of acute toxicity of marine or estuarine sediment to amphipods1ScopeThis International Standard specifies a method for the determination of acute toxicity to amphipods exposedover a period of
toa)samples of contaminated marine or estuarine sediment,b)chemical, industrial or municipal sludge, or other solid wastes that may combine with marine or estuarinesediments, orc)chemicals or preparations spiked into clean sediment.2PrincipleMarine or estuarine amphipods which typically live below the sediment surface are exposed for
tocontaminated sediment or to sediment spiked with a test chemical. The endpoint for the test is percent mortality.The test is performed in 1-litre vessels with
of solid-phase sediment and overlying water. Salinity andtemperature are dependent on the species of amphipod used in testing.3Test environment3.1FacilitiesThe test facility shall be well ventilated, isolated from physical disturbances and free from dust and fumes.3.2LightingAll test vessels shall receive direct, overhead illumination that provides normal laboratory lighting (i.e.
to) at the water surface. Illumination should be uniform and shall be continuous throughout the test periodto inhibit the nocturnal migration of amphipods into the water column[39].4Reagents and materials4.1Test organism4.1.1GeneralOne of the marine or estuarine sediment-dwelling amphipod species listed in AnnexB should be used as testorganism for the method in this International Standard. The species identification should be conducted usingtaxonomic keys[18] and confirmed by a qualified taxonomist familiar with identifying marine or estuarineamphipods.10d10d175ml500lx1000lx



ISO16712:2005(E)2© ISO2005–All rights reserved4.1.2Life stage and sizeAmphipods of uniform age and size shall be used for testing and shall not be larger than the maximum allowablespecies size listed in AnnexB. Do not use mature females bearing embryos, nor individuals longer than themaximum length (including antennae) identified in AnnexB, as they might be senescent.4.1.3SourceAll amphipods used in a test shall be derived from the same population and source. Test organisms can beeither recently collected from an area in which contaminants are at or below background levels, or organismscan be cultured in a laboratory [11], [12], [48].4.1.4Collection, handling and transportDepending on species and/or collection site conditions, collect amphipods using a benthic grab1), a smallbiological dredge or, in inter-tidal zones, a shovel. If a dredge is used, a short haul () minimizes damageto the animals[39]. Collect at least one-third more individuals than are required for the test. Choose a collectionsite for which the presence of abundant organisms of the correct size and age has been demonstratedpreviously, or by pre-collection sieving of the sediment at the site[31]. The organisms to be used as the testspecies should be confirmed taxonomically (e.g. references [4], [5], [32]).Measure and record the salinity, temperature and dissolved oxygen content of water near the sediment at thecollection site. Sieve sediment samples at the time of collection through a sieve of mesh
to . Thechoice of sieve size depends on the size of the species to be collected and is important for determining thenumber of amphipods recovered. The sieve shall be made of non-toxic materials.Use collection-site water for sieving sediment in the field, and to cover the sediment in the container(s) duringspecimen collection and transport. Discard detritus and predators recovered by sieving. Transport the collectedamphipods either at a cool temperature, with only moist inert material such as seaweed in the transportcontainer, or with overlying water and the amphipods returned to the sieved sediment in the transportcontainer(s). Aerate overlying water during transport. Deliver an additional portion of the sieved sediment to thelaboratory to use for holding amphipods and as control sediment. Reserve a portion of sediment for physicalanalysis (e.g. grain-size) and chemical analyses. Alternatively, collect and transport amphipods in bulk sedimentwithout sieving at the field site. However, predatory organisms shall be removed by hand-picking fromcontainers before shipment.Efforts should be made to maintain site-water collection temperature and salinity during transport. Temperaturein the transport container shall not rise above the optimum range for specific amphipod species, as outlined inAnnexB. Overlying seawater shall be aerated during transit.4.1.5Holding and acclimationIf necessary, field-collected specimens may be re-sieved upon return to the laboratory ( to screen, depending on size of amphipods to be used in the test), if one wishes to assess their survival andcondition, to confirm species, and to select and count numbers of amphipods of a size suitable for testing.However, it shall be noted that re-sieving of field-collected amphipods in the laboratory puts an additional stresson the organisms. Use seawater from the collection site, another field site or reconstituted seawater, asoverlying water in the transport container, maintaining the original salinity (within ) and temperature(within
) of the collection-site seawater during transport.In the laboratory, slowly agitate a sieve immersed in seawater to isolate organisms and move them using awide-bore pipette, spoon, or fine net. Ensure that sieved organisms are submersed in seawater at all times. Tominimize stress, handle organisms carefully and quickly. Amphipods that are dropped, injured, or in contact withdry surfaces shall be discarded. Only active and apparently healthy amphipods shall be transferred into the1)Smith-McIntyre and van Veen are examples of suitable products available commercially. This information is given for theconvenience of users of this International Standard and does not constitute an endorsement by ISO of these products.<10m0,5mm1,0mm0,5mm1,0mm±2g/kg±2◦C



ISO16712:2005(E)© ISO2005–All rights reserved3holding/acclimation containers. Depending on the species, any individuals that fail to burrow or that emergefrom the sediment at any time during the holding/acclimation period and appear dead or inactive when gentlyprodded shall be discarded. On the day of a test, select amphipods that are active and apparently healthy, andwhich have an appearance and behaviour typical of that species. Discard any animals that appear or behaveatypically.Count the amphipods selected for use in tests as they are transferred into holding containers (e.g. plastic traysor glass finger bowls). Place at least
to
of previously re-sieved control sediment (free of smallamphipods and other organisms) and at least
to
of overlying seawater in these containers. Thedensity of macroorganisms in the sediment should not exceed either that observed in the field or one amphipodper cm2 to avoid crowding.Place holding containers with organisms in one of the following:a)a tank or trough with flowing seawater;b)a large aquarium (e.g.
to ) containing reconstituted seawater or natural, clean seawater held understatic conditions;c)a smaller aquarium (e.g.
to ) containing seawater held under semi-static conditions (e.g. with dailyrenewal of
of the seawater), unless a recycled water system with proper water treatment is used, inwhich case daily renewal of seawater is not required; ord)a separate room with the appropriate temperature and lighting conditions.A photoperiod of
light and
dark is recommended during amphipod holding/acclimation. The seawater inwhich holding containers are submersed should be aerated.Field-collected or cultured amphipods shall be acclimated to test temperature and salinity conditions for aminimum of . Upon their arrival in the laboratory, acclimate amphipods from the field salinity conditions to thetest salinity conditions by changing the salinity in the holding container at a rate of
(or slowerdepending on the species to be used). Acclimation of amphipods to test temperature conditions, within theholding/acclimation container, should not occur at a rate of temperature increase greater than
per day.Once test salinity conditions have been reached, hold organisms at that salinity for at least
before testing.Temperature, salinity, pH and dissolved oxygen content shall be monitored and recorded daily during the initialacclimation period, when the amphipods are acclimating to the test conditions. Thereafter temperature, salinity,pH and dissolved oxygen content should be measured during the remaining acclimation period, and shall bemeasured and recorded at the end of the acclimation period. Replace the overlying water continuously orperiodically (i.e. daily or every second day) with air-saturated, fresh seawater adjusted to the requiredtemperature and salinity. While the minimum duration of the holding/acclimation period for amphipods is , theholding period for field-collected test organisms shall not exceed
before use in a test. The maximumholding time does not apply to laboratory-cultured test organisms. Amphipods shall not be fed during theirperiod of acclimation or under test conditions.4.2Overlying water4.2.1GeneralAmphipods are to be held and acclimated using either an uncontaminated supply of natural seawater, orreconstituted seawater. The seawater supply used should be monitored and assessed as frequently as requiredto document its quality. Measure the salinity, pH, dissolved oxygen content, ammonia nitrogen, nitrite, relevantpesticides and metals of the seawater used.De-ionized or distilled water is preferred for preparing reconstituted seawater. Dechlorinated municipal drinkingwater, natural surface water or groundwater may also be used.Seawater used for holding, acclimating and testing amphipods shall be free of suspended matter. It isrecommended that seawater be filtered () before use to ensure the removal of suspended particles andorganisms. If stored, hold natural seawater within the range of temperature appropriate for the test species (see2cm4cm2cm5cm60l100l20l40l50%16h8h3d5g/(kg×d)3◦C24h3d14d<5µm



ISO16712:2005(E)4© ISO2005–All rights reservedAnnexB) and use within a few days. For laboratories that have water treatment systems such as sand-bedfilters, seawater may be held for longer periods as long as the quality of the water is closely monitored.Prepare reconstituted seawater by adding hypersaline brine (HSB) or by direct addition of dry salts to a suitablefresh water, in quantities sufficient to provide the desired salinity[22]. HSB may also be prepared usingcommercially available dry ocean salts or reagent-grade salts[47], or by using reagent-grade chemicals toproduce reconstituted salt water (AnnexA). Reconstituted water should be homogeneous and aged for 1weekto 2weeks [1],[2], then filtered () shortly before use to remove suspended particles, and should be usedwithin
of filtration[46].Reconstituted seawater is prepared by adding specified amounts of a suitable salt reagent to high-puritydistilled or de-ionized water[47]. Suitable salt reagents can be reagent grade chemicals or commercial sea salts.Pre-formulated brine (e.g.
to ) prepared with dry ocean salts or heat-concentrated natural seawatercan also be used.4.2.2SalinityThe choice of the appropriate test salinity conditions depends on the salinity of the pore water of the testsediment, the range of salinity tolerance for the test species[10] and the test objectives. For evaluations ofmarine or estuarine sediments, the acclimation and test salinity can range between
and ,depending on the test species chosen (see AnnexB). Salinity can be adjusted by the addition of dry ocean saltsor brine (if too brackish), or distilled water (if too saline).4.2.3Dissolved oxygen contentThe dissolved oxygen content of the seawater overlying the sediment shall be
of the air-saturation valueor higher during the test organism holding or acclimation period, at test initiation and throughout the 10-d test.Maintain this level of dissolved oxygen by gentle aeration of the seawater, using filtered, oil-free compressed air,but the rate of aeration should not suspend the sediment.5ApparatusUse ordinary laboratory apparatus and the following for organism holding or culturing and testing[3], [18], [31], [46].Before initiating a test, ensure all test vessels and associated labware are clean and free of all contaminantsfrom previous use[1], [28].5.1Environmental controls, apparatus to control temperature and light intensity.5.2Measuring apparatus and/or instruments for measuring dissolved oxygen content, pH, salinity, totalorganic carbon, ammonia, nitrate, light intensity and temperature.5.3ContainersContainers and accessories, such as sieves, that might contact the organisms, control or test sediment, andseawater during sorting, handling, holding and acclimation shall be made of non-toxic materials (e.g. glass,stainless steel, polyolefin, nylon, porcelain, polyethylene, polypropylene, fibreglass) cleaned and rinsed withdistilled water, de-ionized water, dechlorinated laboratory water, reconstituted seawater or natural seawaterfrom the collection site or an uncontaminated source.Materials such as copper, zinc, brass, galvanized metal, lead and natural rubber shall not come in contact withthis apparatus and equipment, or with samples of control, reference or test sediment, seawater, or test vessels.1-litre glass containers (beakers or wide-mouthed jars) with internal diameter of approximately
arerecommended for use as test vessels. Cover each vessel with a glass or a plastic lid to reduce the possibility ofcontamination of the contents and to reduce evaporation.5µm24h60%90%1g/kg35g/kg85%10cm



ISO16712:2005(E)© ISO2005–All rights reserved56Treatment and preparation of samples6.1GeneralCollect sediment from reference, control and test sites following established practices[1], [3], [18], [19], [20] or, ifrequired, add a test chemical or preparation to a sample of control sediment[1], [18], [20]. Similar sedimentcollection and handling procedures for both test and reference sediments should be used within the sametesting programme. Store collected sediment in a sealed container in darkness at
until required forthe toxicity test. Drying, freezing and cold storage all affect toxicity and bioavailability of chemicals in sediment.Initiate sediment tests as soon as possible to maintain chemical integrity, but preferably within
and not after unless chemical stability can be assured. Analysis of known chemical contaminants may be conducted onsediment samples from the field, and results may be compared to analysis of sediment at the beginning andend of the test to quantify any changes in chemical concentration or form.6.2Control or reference sedimentControl sediment obtained from the amphipod-collection site can be used as the negative-control sediment fora test, as a clean material for spiking a test chemical, or for organism culture. Clean reference sediment can beused as an additional experimental control.6.3Test sedimentCollect test sediment from the site to be evaluated using apparatus such as coring2) or grab3) devices.Sediment is taken from the middle of the sampler that has not been in contact with the apparatus. Typically, thetop
to
of sediment representing the oxic zone is collected and composited from sufficient samples ofthe site to meet the needs of the test. A deeper depth of sediment may be collected for testing depending on theobjective of the study. Transfer the sediment with a non-reactive, pre-cleaned scoop to an inert vessel, and mixthe composited sample until colour and texture are uniform. Store the composited sample in a clean brownglass container (if organics are suspected contaminants), or in a clean high-density polyethylene orpolycarbonate container (if metals are suspected contaminants). Fill containers to capacity and transport to thelaboratory at .Apparatus should be cleaned between sites to prevent cross-contamination (see Clause5). Retain any solventcleaning wastes and return to the laboratory for disposal.6.4Preparation of sediment samplesRemove large particles () and indigenous organisms by hand sorting using tweezers or a similarinstrument. Sieving the sediment for this purpose is not recommended, because water-soluble contaminantsand fine non-settling clay particles could be lost. Adjust the water to the test temperature and test salinityappropriate for the test species (see AnnexB) and aerate to a dissolved oxygen content of
saturation orhigher. If control/reference sediment is to be used at the completion of the test for determining the ability ofsurviving amphipods to rebury, then re-seal and refrigerate the required sediment.Chemical and physical characterization of the sediment sample is helpful in the interpretation of results. Allowthe sieved sediment to settle for at least
to recover fines, before submitting it for particle size and chemicalanalyses. Analyse a sub-sample of the sediment for the following: total organic carbon and particle sizedistribution (percentage gravel, coarse and fine sands, and silt and clay), and pore water salinity (before sievingin the laboratory). Further characterization can include total volatile residue, acid-volatile sulfides(AVS)/simultaneously extracted metals (SEM), percent water content, biochemical and/or sediment oxygen2)A Phleger box is an example of a suitable product available commercially. This information is given for the convenienceof users of this International Standard and does not constitute an endorsement by ISO of this product.3)Ekman, Ponar, van Veen, Petersen, Shipek and Kajak-Brinkhurst are examples of suitable products availablecommercially. This information is given for the convenience of users of this International Standard and does not constitutean endorsement by IS
...

Questions, Comments and Discussion

Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.