1、September 2011 Translation by DIN-Sprachendienst.English price group 11No 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!$taS“1816248www.din.deDDIN EN ISO 23611-3Soil quality Sampling of soil invertebrates Part 3: Sampling and soil extraction of enchytraeids (ISO 23611-3:2007)English translation of DIN EN ISO 23611-3:2011-09Bodenbeschaffenheit Probenahme von Wirbellosen im Boden Teil 3: Probenahme und Bo
3、denextraktion von Enchytraeen (ISO 23611-3:2007)Englische bersetzung von DIN EN ISO 23611-3:2011-09Qualit du sol Prlvement des invertbrs du sol Partie 3: Prlvement et extraction des enchytrides (ISO 23611-3:2007)Traduction anglaise de DIN EN ISO 23611-3:2011-09SupersedesDIN ISO 23611-3:2007-12www.be
4、uth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 20 pages0 .118 DIN EN ISO 23611-3:2011-09 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee ISO/TC 190 “Soil quality” in collaborati
5、on 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 Committee), Working Committee NA 119-01-02-04 UA Biologische Verfahren of NA 119-0
6、1-02 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: Sampling and extraction of micro-arthropods (Collembola and Acarina) Part 3: Sampl
7、ing 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 programmes with soil invertebrates1) Expert assistance and specialized laborato
8、ries 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 10381-6 DIN ISO 10381-6 ISO 10390 DIN ISO 10390 ISO 10694 DIN ISO 10694 ISO
9、 11272 DIN ISO 11272 ISO 11274 DIN ISO 11274 ISO 11277 DIN ISO 11277 ISO 11461 DIN ISO 11461 ISO 11465 DIN ISO 11465 ISO 14240-1 DIN EN ISO 14240-1 ISO 14240-2 DIN EN ISO 14240-2 ISO 23611-1 DIN EN ISO 23611-1 ISO 23611-2 DIN EN ISO 23611-2 Amendments This standard differs from DIN ISO 23611-3:2007-
10、12 as follows: a) the text of DIN ISO 23611-3:2007-12 has been adopted as European Standard DIN EN ISO 23611-3:2011-09. Previous editions DIN ISO 23611-3: 2007-12 1) In preparation 2 1) DIN EN ISO 23611-3:2011-09 National Annex NA (informative) Bibliography DIN EN ISO 14240-1, Soil quality Determina
11、tion of soil microbial biomass Part 1: Substrate-induced respiration method DIN EN ISO 14240-2, Soil quality Determination of soil microbial biomass Part 2: Fumigation-extraction method DIN EN ISO 23611-1, Soil quality Sampling of soil invertebrates Part 1: Hand-sorting and formalin extraction of ea
12、rthworms DIN EN ISO 23611-2, Soil quality Sampling of soil invertebrates Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina) DIN ISO 10381-1, Soil quality Sampling Part 1: Guidance on the design of sampling programmes DIN ISO 10381-6, Soil quality Sampling Part 6: Guidance o
13、n the collection, handling and storage of soil under aerobic conditions for the assessment of microbiological processes, biomass and diversity in the laboratory DIN ISO 10390, Soil quality Determination of pH DIN ISO 10694, Soil quality Determination of organic and total carbon after dry combustion
14、(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 DIN ISO 11277, Soil quality Determination of particle size distribution in mineral soil material Method by sieving a
15、nd 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 of dry matter and water content on a mass basis Gravimetric method 3 DIN EN ISO 23611-3:2011-09 4 This page is inten
16、tionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 23611-3 July 2011 ICS 13.080.30; 13.080.05 English Version Soil quality - Sampling of soil invertebrates - Part 3: Sampling and soil extraction of enchytraeids (ISO 23611-3:2007) Qualit du sol - Prlvement des invertbrs du sol -
17、Bodenbeschaffenheit - Probenahme von Wirbellosen im Boden - Teil 3: Probenahme und Bodenextraktion von Enchytraeen (ISO 23611-3:2007) 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 f
18、or 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 exists in thr
19、ee 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 standards bo
20、dies 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 and United K
21、ingdom. 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 23611-3:2011
22、: E(ISO 23611-3:2007) Partie 3: Prlvement et extraction des enchytrides Contents Page Foreword. 3 Introduction 4 1 Scope . 5 2 Terms and definitions. 5 3 Principle. 5 4 Reagents 6 5 Apparatus 6 6 Procedure 7 6.1 Soil sampling. 7 6.2 Extraction of the enchytraeids 7 6.3 Microscopic identification . 8
23、 6.4 Preservation of Enchytraeidae 8 6.5 Validity of the extraction process . 8 6.6 Determination of biomass 9 7 Data assessment. 9 8 Test report . 9 Annex A (informative) Species identification in enchytraeids. 11 Annex B (informative) Quick extraction of enchytraeids . 12 Bibliography . 1 2DIN EN
24、ISO 23611-3:2011-09 EN ISO 23611-3:2011 (E) 43DIN EN ISO 23611-3:2011-09 EN ISO 23611-3:2011 (E) Foreword The text of ISO 23611-3:2007 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 2361
25、1-3:2011 by Technical Committee CEN/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 nation
26、al standards shall be withdrawn at 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/CE
27、NELEC Internal Regulations, the national 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, Lithua
28、nia, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 23611-3:2007 has been approved by CEN as a EN ISO 23611-3:2011 without any modification. Introduction This part of ISO 236
29、11 has been drawn up since there is a growing need for the standardization of terrestrial zoological field methods. Such methods, mainly covering the sampling, extraction and handling of soil invertebrates, are needed for the following purposes: g127 biological classification of soils including soil
30、 quality assessment (e.g. References 21, 25, 27); g127 terrestrial bioindication and long-term monitoring (e.g. References 13, 26); g127 evaluation of the effects of chemicals on soil animals (References 15, 22). Data for these purposes are gained by standardized methods since they can form the basi
31、s for far-reaching decisions (e.g. whether a given site should be remediated or not). In fact, the lack of such standardized methods is one of the most important reasons why biological classification concepts in terrestrial (i.e. soil) habitats have so far been relatively rarely used in comparison t
32、o aquatic sites. Originally, the methods described here were developed for taxonomical and ecological studies, investigating the role of enchytraeids in various soil ecosystems. These animals without doubt belong to the most important soil invertebrates in temperate regions (mainly in acidic soils 5
33、). Their influence on soil functions like litter decomposition and nutrient cycling is well-known 14, 19. Due to their number which is often very high (and to their population biomass), they are also important in many terrestrial food-webs 4. Some species have unintentionally been distributed by man
34、 in many soils of the world. Since it is neither possible nor useful to standardize methods for all soil organisms, the most important ones have been selected. Microbiological parameters are already covered by existing ISO guidelines (e.g. ISO 10381-6 29, ISO 14240-1 37and ISO 14240-2 38). 4DIN EN I
35、SO 23611-3:2011-09 EN ISO 23611-3:2011 (E) 1 Scope This part of ISO 23611 specifies a method for sampling, handling and extracting enchytraeids from terrestrial 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). B
36、asic information on the ecology of enchytraeids and their use as bioindicators in the terrestrial environment are included in the Bibliography. This part of ISO 23611 applies to all terrestrial biotopes in which enchytraeids occur. The sampling design of field studies in general is specified in ISO
37、10381-1. These details can vary according to the climatic/regional conditions of the site to be sampled and an overview on the determination of effects of pollutants on enchytraeids in field situations is given in Reference 6. Methods for some other soil organism groups such as earthworms or micro-a
38、rthropods are specified in ISO 23611-1 and ISO 23611-2. This part of ISO 23611 is not applicable for semi-terrestrial (i.e. living in or close to the pure water) soils and might be difficult to use under extreme climatic or geographical conditions (e.g. in high mountains). When sampling soil inverte
39、brates, it is highly recommendable to characterize the site (e.g. concerning climate and land use). However, such a characterization is not covered by this part of ISO 23611. ISO 10390, ISO 10694, ISO 11272, ISO 11274, ISO 11277, ISO 11461 and ISO 11465 are more suitable for measuring pH, particle s
40、ize 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 enchytraeids small soil-inhabiting worms (a few millimetres to several centimetres in length) belonging to the fam
41、ily Enchytraeidae, order Oligochaeta (class Clitellata, phylum Annelida) EXAMPLE Species of the genera Enchytraeus, Fridericia or Cognettia. 3 Principle Enchytraeids at a certain site are sampled from the soil by using a split corer (diameter usually 3 cm to 6 cm). After sampling, the soil samples c
42、ontaining the enchytraeids are transported to the laboratory. Then the enchytraeids are extracted from soil by means of a wet extraction method. (This approach has been well-known for a long time 11, 17, 20.) After extraction, the enchytraeids are identified alive and, if required, preserved in such
43、 a way that they can be stored in a collection indefinitely (e.g. for taxonomical purposes). 5DIN EN ISO 23611-3:2011-09 EN ISO 23611-3:2011 (E) The determination of the biomass of enchytraeids is also described in this part of ISO 23611. The abundance and biomass values can be recalculated to the a
44、rea of the soil corer or, more rarely, volume parameters. NOTE 1 The sampling of enchytraeids is often included in much broader monitoring programmes which try to cover the whole soil fauna or parts of it (e.g. the mesofauna). The design of such programmes is not included in this part of ISO 23611 (
45、but see e.g. Reference 3). NOTE 2 Some hints for the taxonomy of enchytraeids are given in Annex A. 4 Reagents 4.1 Tap water (without toxic properties, e.g. due to copper contamination). 4.2 Ethanol, 70 % (volume fraction). 4.3 Bengalred, 4,5,6,7-Tetrachloro-2,4,5,7-tetraiodofluorescein formulated a
46、s a staining agent. 4.4 Bouins fixative, buffered solution of formaldehyde, acetic acid and picric acid. 4.5 Paracarmin, staining agent, prepared as a mixture of carmine acid, aluminium chloride and calcium chloride solved in ethanol. 4.6 Canada-balm, natural yellowish viscous fluid containing 13 %
47、to 14 % (volume fraction) Canadin acid (C20H38O2), 48 % to 50 % (volume fraction) g68- and g69-Canadinol acid (C19H30O2) and 5 % (volume fraction) Canadoesen (C21H40O). 5 Apparatus 5.1 Split soil corer (e.g. diameter 3 cm to 6 cm; extracted core length 10 cm to 30 cm); length in total variable (depe
48、nding whether or not a handle is used). 5.2 Plastic bags (e.g. 1-l freezer bags); general store. 5.3 Temperature recorder or a minimum/maximum-thermometer. 5.4 Plastic bowls, diameter approximately 20 cm, height approximately 10 cm; general store. 5.5 Plastic sieves, diameter approximately 15 cm, mesh width approximately 0,5 mm; general store. 5.6 60-W bulbs as a heating device; general store. 5.7 Glassware, e.g. petri
