1、BS 6068-5.37:2005, as BS EN ISO 16712:2006)g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58estuarine sediment to amphipodsThe European Standard EN I
2、SO 16712:2006 has the status of a British Standard ICS 13.080.30Water quality Determination of acute toxicity of marine or BRITISH STANDARDBS EN ISO 16712:2006Incorporating amendment no. 1 (renumbers dual numbered with BS ISO 16712:2005, BS EN ISO 16712:2006This British Standard was published under
3、the authority of the Standards Policy and Strategy Committee on 10 March 2005 BSI 2006ISBN 0 580 45592 0Amendments issued since publicationAmd. No. Date Comments16801 29 December 2006 Renumbers BS ISO 16712:2005 as BS EN ISO 16712:2006contract. Users are responsible for its correct application.Compl
4、iance with a British Standard cannot of itself confer immunity from legal obligations.National forewordThis British Standard was published by BSI. It is the UK implementation of EN ISO 16712:2006. It is identical with ISO 16712:2005.The UK participation in its preparation was entrusted by Technical
5、Committee EH/3, Water quality, to Subcommittee EH/3/5, Biological methods.A list of organizations represented on this subcommittee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NOR
6、MEN ISO 16712October 2006ICS 13.080.30English VersionWater quality - Determination of acute toxicity of marine orestuarine sediment to amphipods (ISO 16712:2005)Qualit de leau - Dtermination de la toxicit aigu dessdiments marins et estuariens vis-vis des amphipodes(ISO 16712:2005)Wasserbeschaffenhei
7、t - Bestimmung der akuten Toxizittmariner Sedimente oder von Sedimenten ausFlussmndungsgebieten gegenber 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
8、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
9、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,
10、 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 STANDARDIZATIONC
11、OMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2006 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 16712:2006: EForeword The text of ISO 16712:2005 has been prep
12、ared 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 statu
13、s 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 follo
14、wing 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, Spai
15、n, 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. EN ISO 16712:2006INTERNATIONALSTANDARDISO16712First edition2005-02-15Reference numberISO 16712:2005(E) OSI 2005Water quality Determination
16、 of acute toxicity of marine or estuarine sediment to amphipodsQualit de leau Dtermination de la toxicit aigu des sdiments marins et estuariens vis-vis des amphipodesEN ISO 16712:2006ii ISO 5002 r llAithgs reservde iiiContents Page1 Scope 12 Principle 13 Test environment . 13.1 Facilities . 13.2 Lig
17、hting 14 Reagents and materials 14.1 Test organism 14.2 Overlying water . 35 Apparatus . 46 Treatment and preparation of samples 56.1 General . 56.2 Control or reference sediment . 56.3 Test sediment 56.4 Preparation of sediment samples 57 Test procedures . 67.1 Preparing the exposure vessels 67.2 I
18、ntroducing the organisms . 67.3 Test conditions 77.4 Test observations and measurements 77.5 Expression of results 87.6 Reburrowing capability . 87.7 Validity of the test 88 Analysis and interpretation of results . 88.1 Data analysis . 88.2 Non-contaminant factors 99 Reference toxicant . 910 Precisi
19、on . 911 Test report 10Annex A (informative) Reconstitued salt water . 11Annex B (normative) Maximum length of amphipod species, optimal salinity and temperatureranges . 12Annex C (informative) Precision data . 13Bibliography . 14EN ISO 16712:2006iv ISO 5002 All rithgs reresvdeForewordISO (the Inter
20、national 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 be
21、enestablished 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 standar
22、dization.International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.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 voti
23、ng. 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 pa
24、tent rights.ISO 16712 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biologicalmethods.EN ISO 16712:2006 ISO 5002 r llAithgs reservde vIntroductionThis International Standard outlines procedures for conducting acute tests for sediment toxicity, using one ormore amp
25、hipod 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.EN ISO 16712:2006blankANRETNIITOTS LANDNADRA ISO 5002 r llAithgs reservde 1Water quality Determination of acute toxicity of mar
26、ine 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 ma
27、y combine with marine or estuarinesediments, orc) chemicals or preparations spiked into clean sediment.2 PrincipleMarine 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 tes
28、t 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.3 Test environment3.1 FacilitiesThe test facility shall be well ventilated, isolated from physical distur
29、bances and free from dust and fumes.3.2 LightingAll 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
30、 amphipods into the water column39.4 Reagents and materials4.1 Test organism4.1.1 GeneralOne of the marine or estuarine sediment-dwelling amphipod species listed in Annex B should be used as testorganism for the method in this International Standard. The species identification should be conducted us
31、ingtaxonomic keys18and confirmed by a qualified taxonomist familiar with identifying marine or estuarineamphipods.10 d10 d175 ml500 lx1 000 lxEN ISO 16712:20062 ISO 5002 All rithgs reresvde4.1.2 Life stage and sizeAmphipods of uniform age and size shall be used for testing and shall not be larger th
32、an the maximum allowablespecies size listed in Annex B. Do not use mature females bearing embryos, nor individuals longer than themaximum length (including antennae) identified in Annex B, as they might be senescent.4.1.3 SourceAll amphipods used in a test shall be derived from the same population a
33、nd 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 11,12,48.4.1.4 Collection, handling and transportDepending on species and/or collection site conditions, collect amphipods using
34、 a benthic grab1), a smallbiological dredge or, in inter-tidal zones, a shovel. If a dredge is used, a short haul ( ) minimizes damageto the animals39. Collect at least one-third more individuals than are required for the test. Choose a collectionsite for which the presence of abundant organisms of
35、the correct size and age has been demonstratedpreviously, or by pre-collection sieving of the sediment at the site31. 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
36、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-to
37、xic 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
38、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
39、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 shipm
40、ent.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 inAnnex B. Overlying seawater shall be aerated during transit.4.1.5 Holding
41、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 s
42、uitable 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
43、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
44、 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
45、 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.1cm85 %4hEN ISO 16712:20066 ISO 5002 All rithgs reresvdedemand, total metals, total chlorinated organic content, chlorinated o
46、rganic compounds, polycyclic aromatichydrocarbons, and oil and grease3,18. Pore water can also be sampled and analysed for pH, total ammonia,hydrogen sulfide and dissolved contaminants. Samples of pore water can be obtained from control or referencesediment by in situ methods (e.g. peepers) or ex si
47、tu methods (e.g. centrifugation or squeezing)1, 19, 35.Mix each sediment sample thoroughly the day preceding the test and add a aliquot of sediment to a testvessel (see 6.3). Distribute test sediment in a uniform layer, which allows amphipods to burrow (minimum in depth). Prepare a minimum of three
48、laboratory replicates for each treatment or test sample, plus one or morereplicate treatments for monitoring chemical characteristics of the sediment and the test water during the test.Replicates set up for monitoring of chemistry shall be seeded with amphipods, as in test replicates.Smooth the sedi
49、ment surface flat in each treatment and ensure there is minimal disturbance of the test orcontrol sediment during the addition of overlying water. One technique for minimizing the disruption of sedimentis to add the seawater by pouring over a disk (polyethylene, nylon or polytetrafluoroethylene sheeting of to thickness) lying on the sediment surface that fits the inside diameter of the test vessel3, 18. Addseawater to each test vessel to make up a total (water plus sediment) volume of .
