1、September 2011 Translation by DIN-Sprachendienst.English price group 12No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).
2、ICS 13.080.30!$taR“1816247www.din.deDDIN EN ISO 23611-2Soil quality Sampling of soil invertebrates Part 2: Sampling and extraction of micro-arthropods (Collembola andAcarina) (ISO 23611-2:2006)English translation of DIN EN ISO 23611-2:2011-09Bodenbeschaffenheit Probenahme von Wirbellosen im Boden Te
3、il 2: Probenahme und Extraktion von Mikroarthropoden (Collembolen und Milben)(ISO 23611-2:2006)Englische bersetzung von DIN EN ISO 23611-2:2011-09Qualit du sol Prlvement des invertbrs du sol Partie 2: Prlvement et extraction des micro-arthropodes (Collembola et Acarina)(ISO 23611-2:2006)Traduction a
4、nglaise de DIN EN ISO 23611-2:2011-09SupersedesDIN ISO 23611-2:2007-02www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 0 pages0 .1182DIN EN ISO 23611-2:2011-09 A comma is used as the decimal marker. National foreword This standard has be
5、en prepared by Technical Committee ISO/TC 190 “Soil quality” in collaboration with Technical Committee CEN/TC 345 “Characterization of soils” (Secretariat: NEN, Netherlands). The responsible German body involved in its preparation was the Normenausschuss Wasserwesen (Water Practice Standards Committ
6、ee), Working Committee NA 119-01-02-04 UA Biologische Verfahren of NA 119-01-02 AA Abfall- und Bodenuntersuchung. 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: Samplin
7、g and extraction of micro-arthropods (Collembola and Acarina) Part 3: Sampling and soil extraction of enchytraeids Part 4: Sampling, extraction and identification of soil-inhabiting nematodes Part 5: Sampling and extraction of soil macro-invertebrates Part 6: Guidance for the design of sampling prog
8、rammes with soil invertebrates1) Expert assistance and specialized laboratories will be required to perform the sampling procedures described in this standard. The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 10381-1 DIN ISO 10381-1 ISO
9、10390 DIN ISO 10390 ISO 10694 DIN ISO 10694 ISO 11272 DIN ISO 11272 ISO 11274 DIN ISO 11274 ISO 11277 DIN ISO 11277 ISO 11461 DIN ISO 11461 ISO 11465 DIN ISO 11465 Amendments This standard differs from DIN ISO 23611-2:2007-02 as follows: a) the text of DIN ISO 23611-2:2007-02 has been adopted as Eur
10、opean Standard DIN EN ISO 23611-2:2011-09. Previous editions DIN ISO 23611-2: 2007-02 1) In preparation 2 1) DIN EN ISO 23611-2:2011-09 3 National Annex NA (informative) Bibliography DIN ISO 10381-1, Soil quality Sampling Part 1: Guidance on the design of sampling programmes DIN ISO 10390, Soil qual
11、ity Determination of pH DIN ISO 10694, Soil quality Determination of organic and total carbon after dry combustion (elementary analysis) DIN ISO 11272, Soil quality Determination of dry bulk density DIN ISO 11274, Soil quality Determination of the water retention characteristics Laboratory methods D
12、IN ISO 11277, Soil quality Determination of particle size distribution in mineral soil material Method by sieving and sedimentation DIN ISO 11461, Soil quality Determination of soil water content as a volume fraction using coring sleeves Gravimetric method DIN ISO 11465, Soil quality Determination o
13、f dry matter and water content on a mass basis Gravimetric method This page is intentionally blank DIN EN ISO 23611-2:2011-09 4EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 23611-2 July 2011 ICS 13.080.30; 13.080.05 English Version Soil quality - Sampling of soil invertebrates - Part 2: S
14、ampling and extraction of micro-arthropods (Collembola and Acarina) (ISO 23611-2:2006) Qualit du sol - Prlvement des invertbrs du sol - (Collembola et Acarina) (ISO 23611-2:2006) Bodenbeschaffenheit - Probenahme von Wirbellosen im Boden - Teil 2: Probenahme und Extraktion von Mikroarthropoden (Colle
15、mbolen und Milben) 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 standard without any alteration. Up-to-date lists and bib
16、liographical 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 respo
17、nsibility 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 Republic, Denmark, Estonia, Finland, France, Germany, Gr
18、eece, 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 NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Manage
19、ment 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 23611-2:2011: EPartie 2: Prlvement et extraction des micro-arthropodes (ISO 23611-2:2006) Contents Page Foreword. 3 Introductio
20、n 4 1 Scope . 5 2 Terms and definitions. 5 3 Principle. 5 4 Test materials 6 4.1 Biological material 6 4.2 Reagents 6 5 Apparatus 7 6 Procedure 8 6.1 Collecting the soil samples . 8 6.2 Extracting Collembola and Acarina from soil samples 8 6.3 Sorting, preserving and identifying Collembola and Acari
21、na 9 7 Assessment of results 10 8 Study report. 10 Annex A (informative) Species determination in collembolans and mites 11 Annex B (informative) Alternative methods for sampling of micro-arthropods 13 Bibliography . 1 2DIN EN ISO 23611-2:2011-09 EN ISO 23611-2:2011 (E) 4Foreword The text of ISO 236
22、11-2:2006 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/TC 345 “Characterization of soils” the secretariat of which is held by NEN. This European
23、 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 of the element
24、s 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 implement this Europ
25、ean 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, Switzerland and th
26、e 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. 3DIN EN ISO 23611-2:2011-09 EN ISO 23611-2:2011 (E) Introduction This part of ISO 23611 has been drawn up since there is a growing need for the standardizatio
27、n of sampling and extraction methods of soil micro-arthropods. These methods are needed for the following purposes: g127 biological classification of soils including soil quality assessment (e.g. References 31, 32, 35, 41, 45, 46); g127 terrestrial bioindication and long-term monitoring (e.g. Refere
28、nces 1, 7, 17, 40, 42). Data collected by standardized methods can be more accurately evaluated allowing more reliable comparisons between sites (e.g. polluted versus non-polluted sites, changes in land-use practices). From the several micro-arthropod groups, Collembola and Acarina are the most stud
29、ied in soil ecology. Their relevance for the soil system comes from their high abundance and diversity, and also from their role in key biological processes. Collembola and Oribatid mites act mainly as catalysts in organic matter decomposition 4, 20, whereas predacious mites may act as webmasters in
30、 soil food webs 9. These characteristics, allied to a widespread taxonomic knowledge, allowed their use as study organisms in several research programmes 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)
31、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 quality, especially due to land-use practices and pollution 43. 4DIN EN ISO 23611-2:2011-09 EN ISO 23611-2:2011 (E) 1 Scope This part of ISO 23611 specifies
32、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-arthropods and their use can be found in t
33、he 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 than soil, e.g. tree trunks, plants or lich
34、ens. 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 site which is highly recommendable when sam
35、pling 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 the purposes of this document, the follow
36、ing 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 composed by mites (Acarina), springtails (Col
37、lembola), 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 cores are placed in plastic tubes (or plas
38、tic 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 are also described. Finally, abundance va
39、lues can be recalculated related to area (usually 1 m2), volume or weight (usually 1 kg). 5DIN EN ISO 23611-2:2011-09 EN ISO 23611-2:2011 (E) NOTE Alternative methods for extraction can be used under special circumstances. Flotation methods (e.g. the heptane flotation method) can be used in clay or
40、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 with a pair of antennae, without true compound eyes, with six abd
41、ominal 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 = glue and embolon = bar). In the third segment, there is the
42、 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 forms. They belong to the class Collembola, and can be separated
43、 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 Acarida, and can be separated into four groups: Cryptostigmata
44、 (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 and distilled water. 4.2.1 Propan-2-ol, 80 % (volume fraction)
45、. 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 hydrate). 4.2.7 1,2,3-Trihydroxypropane (glycerine). 4.2.8 von
46、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 chloral hydrate (80 g), distilled water (50 ml) and concentrated
47、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 micro-arthropod specimens, especially oribatid mites under th
48、e 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). 6DIN EN ISO 23611-2:2011-09 EN ISO 23611-2:2011 (E) 4.2.13 Hoyers medium, used to mount Collembola specimens composed of distilled water (50 ml),
49、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 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 l
