1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN ISO 23611-1:2011Soil quality Sampling of soilinvertebratesPart 1: Hand-sorting and formalinextraction of earthworms (ISO23611-1:2006)BS EN ISO 23611-1:2011 BRITISH STANDARD
2、National forewordThis British Standard is the UK implementation of EN ISO23611-1:2011. It is identical to ISO 23611-1:2006.The UK participation in its preparation was entrusted to TechnicalCommittee EH/4, Soil quality.A list of organizations represented on this committee can beobtained on request to
3、 its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. BSI 2011ISBN 978 0 580 73162 4ICS 13.080.05; 13.080.30Compliance with a British Standard cannot confer immunity fromlegal obligations.This British
4、Standard was published under the authority of theStandards Policy and Strategy Committee on 31 July 2011.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 23611-1 July 2011 ICS 13.080.30; 13.080.05 English Version Soil quality - Sampling of
5、 soil invertebrates - Part 1: Hand-sorting and formalin extraction of earthworms (ISO 23611-1:2006) Qualit du sol - Prlvement des invertbrs du sol - Partie 1 : Tri manuel et extraction au formol des vers de terre (ISO 23611-1:2006) Bodenbeschaffenheit - Probenahme von Wirbellosen im Boden - Teil 1:
6、Handauslese und Formalinextraktion von Regenwrmern (ISO 23611-1:2006) This European Standard was approved by CEN on 17 June 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 standa
7、rd 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
8、 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 Repub
9、lic, 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 STANDARDIZATION COMIT EUROPEN DE
10、NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2011 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 23611-1:2011: EBS EN ISO 23611-1:2011EN ISO 23611-1:2011 (E) 3 Foreword The
11、text of ISO 23611-1:2006 has been prepared by Technical Committee ISO/TC 190 “Soil quality” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 23611-1:2011 by Technical Committee CEN/TC 345 “Characterization of soils” the secretariat of which is held by NEN
12、. 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 January 2012, and conflicting national standards shall be withdrawn at the latest by January 2012. Attention is drawn to the possibility that some
13、 of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to imple
14、ment this European Standard: Austria, Belgium, Bulgaria, Croatia, 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, Swi
15、tzerland and the United Kingdom. Endorsement notice The text of ISO 23611-1:2006 has been approved by CEN as a EN ISO 23611-1:2011 without any modification. BS EN ISO 23611-1:2011ISO 23611-1:2006(E) ISO 2006 All rights reserved iiiContents Page Foreword iv Introduction v 1 Scope . 1 2 Terms and defi
16、nitions. 1 3 Principle. 2 4 Reagents 2 5 Apparatus 2 6 Procedure 3 6.1 Sampling of the earthworms . 3 6.2 Preservation 5 6.3 Determination of biomass 5 7 Data assessment. 5 8 Study report. 6 Annex A (informative) Other methods for sampling. 7 Annex B (informative) Species determination in peregrine
17、earthworms . 8 Annex C (informative) The modified TSBF Method 9 Bibliography . 10 BS EN ISO 23611-1:2011ISO 23611-1:2006(E) iv ISO 2006 All rights reservedForeword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The
18、 work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-gove
19、rnmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. T
20、he main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vot
21、e. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. ISO 23611-1 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biolo
22、gical methods. ISO 23611 consists of the following parts, under the general title Soil quality Sampling of soil invertebrates: Part 1: Hand-sorting and formalin extraction of earthworms Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina) Part 3: Sampling and soil extraction
23、of enchytraeids Part 4: Sampling, extraction and identification of free-living stages of terrestrial nematodes BS EN ISO 23611-1:2011ISO 23611-1:2006(E) ISO 2006 All rights reserved vIntroduction This part of ISO 23611 has been drawn up since there is a growing need for the standardisation of terres
24、trial zoological field methods. Such methods, mainly covering the sampling, extraction and handling of soil invertebrates, are necessary for the following purposes: biological classification of soils including soil quality assessment 21, 26, 34; terrestrial bioindication and long-term monitoring9, 1
25、2, 28; evaluation of the effects of chemicals on soil animals (ISO 11268-3). Data for these purposes are gained by standardized methods since they can form the basis for far-reaching decisions (e.g. whether a given site should be remediated or not). In fact, the lack of such standardised methods is
26、one of the most important reasons why bio-classification and bio-assessment in terrestrial (i.e. soil) habitats has so far relatively rarely been used in comparison to aquatic sites. Originally, the methods described here were developed for taxonomical and ecological studies, investigating the role
27、of earthworms in various soil ecosystems. These animals are without doubt the most important soil invertebrates in temperate regions and, to a lesser extent, in tropical soils 25, 13, 15. Since Darwin (1881), their influence on soil structure (e.g. aeration, water holding capacity) and soil function
28、s like litter decomposition and nutrient cycling is well-known 8. Due to their often very high biomass they are also important in many terrestrial food-webs. Since it is neither possible nor useful to standardize methods for all soil organisms, the most important ones have been selected. BS EN ISO 2
29、3611-1:2011BS EN ISO 23611-1:2011INTERNATIONAL STANDARD ISO 23611-1:2006(E) ISO 2006 All rights reserved 1Soil quality Sampling of soil invertebrates Part 1: Hand-sorting and formalin extraction of earthworms 1 Scope This part of ISO 23611 specifies a method for sampling and handling earthworms from
30、 field soils as a prerequisite for using these animals as bioindicators (e.g. to assess the quality of a soil as a habitat for organisms). Basic information on the ecology of earthworms and their use as bioindicators in the terrestrial environment can be found in the references listed in the bibliog
31、raphy. This part of ISO 23611 applies to all terrestrial biotopes in which earthworms occur. The sampling design of field studies in general is specified in ISO 10381-1 (see also Reference 38) and guidance on the determination of effects of pollutants on earthworms in field situations is given in IS
32、O 11268-3. These details can vary according to the national requirements or the climatic/regional conditions of the site to be sampled (see also Annex C). This part of ISO 23611 is not applicable for semi-terrestrial soils and it can be difficult to use under extreme climatic or geographical conditi
33、ons (e.g. in high mountains). Methods for some other soil organism groups, such as collembolans, are covered in other parts of ISO 23611. This part of ISO 23611 does not cover the pedological characterization of the site which is highly recommendable when sampling soil invertebrates. ISO 10390, ISO
34、10694, ISO 11272, ISO 11274, ISO 11277, ISO 11461 and ISO 11465 are more suitable for measuring pH, particle size distribution, C/N ratio, organic carbon content and water-holding capacity. 2 Terms and definitions For the purposes of this document, the following terms and definitions apply. 2.1 eart
35、hworm megadrile soil-inhabiting earthworms (length of adult individuals: few centimetres to more than 1 m) belonging to the order Oligochaeta (class Clitellata, phylum Annelida) EXAMPLE Species of the families Lumbricidae (Holarctic), Glossoscolecidae (Latin America), Eudrilidae (Africa) or Megascol
36、ecidae Asia, North America (Pacific Coast). 2.2 peregrine species earthworms occurring in many regions world-wide today, usually introduced by man NOTE 1 Well-known examples of peregrine species are several lumbricid species like Aporrectodea caliginosa (originally coming from Eurasia, but now livin
37、g also in the Americas and Australia) or the pan-tropical species Pontoscolex corethrurus (probably coming from Northern Brazil and/or the Guyanas). NOTE 2 See Reference 15. BS EN ISO 23611-1:2011ISO 23611-1:2006(E) 2 ISO 2006 All rights reserved2.3 clitellum ring or saddle of epidermal thickening o
38、nly in mature worms which is near the anterior and eventually forms the cocoon 3 Principle Earthworms at a certain site are sampled from the soil by using a combination of two different methods: hand-sorting animals from a certain area (e.g. 0,25 m2) or volume (e.g. 50 cm 50 cm 20 cm); extraction of
39、 worms from the soil by applying formalin. Both methods are well-known for a long time 5,20. After extraction, the earthworms are fixed and transported to the laboratory. There they are preserved in a way that they can be stored in a collection indefinitely (e.g. for taxonomical purposes). In additi
40、on, the determination of the biomass of earthworms is described. Finally, abundance and biomass values can be recalculated to area (usually 1 m2) or, more rarely, volume parameters. NOTE 1 Alternative methods can be useful under special circumstances (e.g. electrical extraction or the use of mustard
41、 as a chemical expellent), but cannot be recommended as a general procedure (see Annex A). In addition, the use of other extraction chemicals than formalin seems to have no advantages concerning extraction efficiency 11; 33. NOTE 2 The sampling of earthworms is often included in much broader monitor
42、ing programs, trying to cover the whole soil fauna or parts of it (e.g. the macrofauna). The design of such programs is not included in this part of ISO 236111. NOTE 3 Some hints for the taxonomy of peregrine (occurring in many regions world-wide) earthworms, mainly belonging to the family Lumbricid
43、ae, are given in Annex B. 4 Reagents 4.1 Formalin formaldehyde solution 4 % (volume fraction). 4.2 Formalin formaldehyde solution 37 % (volume fraction). 4.3 Ethanol, 70 % (volume fraction). 5 Apparatus Use standard laboratory equipment and the following. 5.1 Plastic vessels, capacities 250 ml and 5
44、00 ml, for storing the worms. 5.2 Plastic gloves. 5.3 Forceps. 5.4 Piece of thick plastic sheeting, 1 m2to 2 m2. 5.5 Spade or shovel. 5.6 Dissecting microscope, with low magnification (10 to 40 times). 5.7 Balance, able to weigh from 0,01 g to 200 g. 5.8 Water-can, preferably 20 l, with water (20 l
45、per sampling plot). BS EN ISO 23611-1:2011ISO 23611-1:2006(E) ISO 2006 All rights reserved 35.9 Watering can. 5.10 Pencil, notebook, water resistant marker, labels that go in the vessel. 5.11 Thermometer, e.g. for measuring air temperature. 5.12 Drying cabinet, for soil moisture determination. 6 Pro
46、cedure 6.1 Sampling of the earthworms 6.1.1 General Sampling of earthworms is done by a combination of two different methods: hand-sorting and formalin extraction. Based on several comparative studies, this combination is clearly recommended in the various reviews on earthworm ecology (e.g. Referenc
47、es 7, 8, 15). Sampling should be done at times of the year where the animals are not forced by the environmental conditions (i.e. low soil moisture and/or high temperatures) into diapause (i.e. are not reacting to formalin). In temperate regions, such unfavourable sampling times are winter and, in p
48、articular, midsummer periods 15. Earthworms sampled from the same plot, but sampled under the two different methods, should be stored in individual plastic vessels. After the end of the sampling process, the excavated and examined soil is returned to the original sampling plot. In some cases, it is
49、appropriate to use only one of the two methods; e.g. when no deep-burrowing animals are occurring at a given site, formalin extraction is not necessary. On the other hand, at sites where giant earthworms are living (parts of South America, South East Asia and Australia), hand-sorting is not useful 22. A very similar method, known as modified TSBF method, is particularly suitable for tropical regions (see Annex C). NOTE Usually the earthworms are determined after preservation
