1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN ISO 23611-2:2011Soil quality Sampling of soilinvertebratesPart 2: Sampling and extraction of micro-arthropods (Collembola and Acarina) (ISO23611-2:2006)BS EN ISO 23611-2:20
2、11 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO23611-2:2011. It is identical to ISO 23611-2:2006.The UK participation in its preparation was entrusted to TechnicalCommittee EH/4, Soil quality.A list of organizations represented on this committee can beobt
3、ained on request to 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 73163 1ICS 13.080.05; 13.080.30Compliance with a British Standard cannot confer immunity fromlegal obliga
4、tions.This British 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 23611-2 July 2011 ICS 13.080.30; 13.080.05 English Version Soil qu
5、ality - Sampling of soil invertebrates - Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina) (ISO 23611-2:2006) Qualit du sol - Prlvement des invertbrs du sol - Partie 2 : Prlvement et extraction des micro-arthropodes (Collembola et Acarina) (ISO 23611-2:2006) Bodenbeschaffe
6、nheit - Probenahme von Wirbellosen im Boden - Teil 2: Probenahme und Extraktion von Mikroarthropoden (Collembolen und Milben) (ISO 23611-2:2006) 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 c
7、onditions 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 ex
8、ists 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 s
9、tandards bodies of 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, Switzerland a
10、nd United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE 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 2
11、3611-2:2011: EBS EN ISO 23611-2:2011EN ISO 23611-2:2011 (E) 3 Foreword The text of ISO 23611-2:2006 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 23611-2:2011 by Technical Committee CEN
12、/TC 345 “Characterization of soils” the secretariat of which is held by NEN. 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
13、the latest by January 2012. Attention is drawn to the possibility that some 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 nat
14、ional standards organizations of the following countries are bound to implement 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
15、, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 23611-2:2006 has been approved by CEN as a EN ISO 23611-2:2011 without any modification. BS EN ISO 23611-2:2011ISO 23611-2:2006(E) ISO 2006 All rights reserv
16、ed iiiContents Page Foreword iv Introduction v 1 Scope . 1 2 Terms and definitions. 1 3 Principle. 1 4 Test materials 2 4.1 Biological material 2 4.2 Reagents 2 5 Apparatus 3 6 Procedure 4 6.1 Collecting the soil samples . 4 6.2 Extracting Collembola and Acarina from soil samples 4 6.3 Sorting, pres
17、erving and identifying Collembola and Acarina 5 7 Assessment of results 6 8 Study report. 6 Annex A (informative) Species determination in collembolans and mites 7 Annex B (informative) Alternative methods for sampling of micro-arthropods 9 Bibliography . 10 BS EN ISO 23611-2:2011ISO 23611-2:2006(E)
18、 iv ISO 2006 All rights reservedForeword ISO (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 ISO technical committees. Each member body inter
19、ested 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-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical
20、 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. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the te
21、chnical 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 vote. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. IS
22、O shall not be held responsible for identifying any or all such patent rights. ISO 23611-2 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods. ISO 23611 consists of the following parts, under the general title Soil quality Sampling of soil invertebrat
23、es: 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 of enchytraeids Part 4: Sampling, extraction and identification of free-living stages of terrestrial nematodes BS EN ISO 23
24、611-2:2011ISO 23611-2:2006(E) ISO 2006 All rights reserved vIntroduction This part of ISO 23611 has been drawn up since there is a growing need for the standardization of sampling and extraction methods of soil micro-arthropods. These methods are needed for the following purposes: biological classif
25、ication of soils including soil quality assessment (e.g. References 31, 32, 35, 41, 45, 46); terrestrial bioindication and long-term monitoring (e.g. References 1, 7, 17, 40, 42). Data collected by standardized methods can be more accurately evaluated allowing more reliable comparisons between sites
26、 (e.g. polluted versus non-polluted sites, changes in land-use practices). From the several micro-arthropod groups, Collembola and Acarina are the most studied in soil ecology. Their relevance for the soil system comes from their high abundance and diversity, and also from their role in key biologic
27、al processes. Collembola and Oribatid mites act mainly as catalysts in organic matter decomposition 4, 20, whereas predacious mites may act as webmasters in soil food webs 9. These characteristics, allied to a widespread taxonomic knowledge, allowed their use as study organisms in several research p
28、rogrammes dealing with the impacts of forest practices (e.g. References 12, 13, 14, 15, 18, 19, 21, 22, 23, 25, 26, 27, 28, 29, 30, 31, 33, 34, 37, 38, 39) or crop management practices (e.g. 6, 11, 16, 24). These features make them suitable organisms to be used as bio-indicators of changes in soil q
29、uality, especially due to land-use practices and pollution 43. BS EN ISO 23611-2:2011BS EN ISO 23611-2:2011INTERNATIONAL STANDARD ISO 23611-2:2006(E) ISO 2006 All rights reserved 1Soil quality Sampling of soil invertebrates Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina)
30、 1 Scope This part of ISO 23611 specifies a method for sampling, extracting and preserving collembolans and mites from field soils as a prerequisite for using these animals as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms). Basic information on the ecology of micro-
31、arthropods and their use can be found in the references listed in the Bibliography. The sampling and extraction methods of this part of ISO 23611 are applicable to almost all types of soils. Exceptions may be soils from extreme climatic conditions (hard, frozen or flooded soils) and other matrices t
32、han soil, e.g. tree trunks, plants or lichens. For the sampling design of field studies in general, see ISO 10381-1. Methods for some other soil organism groups such as earthworms are covered in other parts of ISO 23611. This part of ISO 23611 does not cover the pedological characterization of the s
33、ite which is highly recommendable when sampling soil invertebrates. ISO 10390, ISO 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
34、 the purposes of this document, the following terms and definitions apply. 2.1 micro-arthropods group which is defined by its small size (range size from 100 m to a few millimetres) making up a significant part of the below-ground food web in many terrestrial ecosystems NOTE This group is mainly com
35、posed by mites (Acarina), springtails (Collembola), Protura, Diplura, garden centipedes (Symphyla), Pauropoda, small centipedes and millipedes, and insects and their larvae from several orders (Diptera, Coleoptera, etc.). 3 Principle Soil samples are collected in the field using a split corer. Soil
36、cores are placed in plastic tubes (or plastic bags) and transported to the laboratory. Afterwards, Collembola and Acarida are rapidly (within a few days) extracted by behavioural methods, using a MacFadyen apparatus, and preserved for future identifications 7, 40. In addition, preparation techniques
37、 are also described. Finally, abundance values can be recalculated related to area (usually 1 m2), volume or weight (usually 1 kg). BS EN ISO 23611-2:2011ISO 23611-2:2006(E) 2 ISO 2006 All rights reservedNOTE Alternative methods for extraction can be used under special circumstances. Flotation metho
38、ds (e.g. the heptane flotation method) can be used in clay or loamy soils and a Kempson extractor is advisable in the case litter is sampled 40. 4 Test materials 4.1 Biological material Collembola (springtails) are small wingless hexapods (from 150 m up to 9 mm length), having a distinctive head wit
39、h a pair of antennae, without true compound eyes, with six abdominal segments and three pre-genital appendages in the abdomen. In the first segment, there is the ventral tube (or collophore) that is used for adhering to smooth surfaces. The name Collembola comes from this structure (from Greek colla
40、 = glue and embolon = bar). In the third segment, there is the tenaculum, that holds the jumping apparatus on its normal position. This jumping appendage, the furcula (or spring), when it exists, is located in the fourth segment. Springtails live in litter and soil, and have very distinctive life fo
41、rms. They belong to the class Collembola, and can be separated into 18 families 17. Soil mites are small chelicerate arthropods related to spiders (length from 150 m up to 5 mm), living in soil and litter, and also presenting very distinctive life forms. They belong to the class Arachnida, subclass
42、Acarida, and can be separated into four groups: Cryptostigmata (Oribatida), Mesostigmata (Gamasida), Prostigmata (Trombidiformes) and Astigmata. NOTE Some hints for the taxonomy of springtails and mites are given in Annex A. 4.2 Reagents Unless otherwise specified, use only reagents of good quality
43、and distilled water. 4.2.1 Propan-2-ol, 80 % (volume fraction). 4.2.2 Formalin formaldehyde solution 40 % (volume fraction). 4.2.3 Acetic acid. 4.2.4 Phenol, C6H5OH, crystalline (carbolic acid). 4.2.5 Hydrogen chloride, c(HCl) from 8 mol/l to 10 mol/l. 4.2.6 2,2,2-Trichloro-1,1-ethanediol (chloral h
44、ydrate). 4.2.7 1,2,3-Trihydroxypropane (glycerine). 4.2.8 von Trne fixative, used to preserve the extracted animals and composed by Propan-2-ol (80 %), formalin (40 %) and glacial acetic acid (a volume fraction 10:0,3:0,03). 4.2.9 Nesbitt clearing medium, used to clear mite specimens composed of chl
45、oral hydrate (80 g), distilled water (50 ml) and concentrated hydrogen chloride (5 ml). 4.2.10 Lactophenol solution, used to clear mite specimens composed of lactic acid (10 ml), crystals of phenol (3,6 g) and distilled water (5 ml). 4.2.11 2-Hydroxypropanoic acid (lactic acid), to clear and observe
46、 micro-arthropod specimens, especially oribatid mites under the microscope. 4.2.12 Ethanol, 70 % to 75% (volume fraction), used for fixation and preservation (in this case, also in combination with glycerine, 10:1). BS EN ISO 23611-2:2011ISO 23611-2:2006(E) ISO 2006 All rights reserved 34.2.13 Hoyer
47、s medium, used to mount Collembola specimens composed of distilled water (50 ml), gum-arabic (30 g), chloral hydrate (200 g) and glycerine (20 ml). 5 Apparatus Use standard laboratory equipment and the following. 5.1 Measuring tape. 5.2 Collecting flasks. 5.3 Wash bottle. 5.4 Forceps, pipette, fine
48、painting brush, fine needles. 5.5 Petri dishes. 5.6 Stereomicroscope. 5.7 Microscope, with phase or interference contrast is preferable. 5.8 Microscopic slides, with excavated area in the centre, and lamellae. 5.9 Electrical heating plate. 5.10 Plastic vials. 5.11 Ceramic heating elements. 5.12 Penc
49、il, notebook, water resistant marker, labels. 5.13 Split corer Sampling device made of stainless steel or aluminium (40 cm long and e.g. 5,6 cm diameter may be used; the size and diameter should not differ considerably from these numbers in order to maintain comparable conditions), used to collect soil cores (samples). It can be composed of two independent parts that fit together along the corer main axis or it can consist of one tube. On
