1、September 2011 Translation by DIN-Sprachendienst.English price group 13No 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!$ta=“1816226www.din.deDDIN EN ISO 23611-4Soil quality Sampling of soil invertebrates Part 4: Sampling, extraction and identification of soil-inhabitingnematodes (ISO 23611-4:2007)English translation of DIN EN ISO 23611-4:2011-09Bodenbeschaffenheit Probenahme von Wirbellosen im Boden Te
3、il 4: Probenahme, Extraktion und Bestimmung von Boden bewohnenden Nematoden(ISO 23611-4:2007)Englische bersetzung von DIN EN ISO 23611-4:2011-09Qualit du sol Prlvement des invertbrs du sol Partie 4: Prlvement, extraction et identification des nmatodes du sol(ISO 23611-4:2007)Traduction anglaise de D
4、IN EN ISO 23611-4:2011-09SupersedesDIN ISO 23611-4:2008-11www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises pages0 .11827DIN EN ISO 23611-4:2011-09 A comma is used as the decimal marker. National foreword This standard has been prepared b
5、y 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 Committee), Working
6、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: Sampling and extract
7、ion 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 programmes with s
8、oil 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 10381-6 DIN I
9、SO 10381-6 ISO 10390 DIN ISO 10390 ISO 10694 DIN ISO 10694 ISO 11268-3 DIN ISO 11268-3 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 23611-4:2008-11 as follows: a) the text of DIN
10、ISO 23611-4:2008-11 has been adopted as European Standard DIN EN ISO 23611-4:2011-09. Previous editions DIN ISO 23611-4: 2008-11 1) In preparation 2 1) DIN EN ISO 23611-4:2011-09 National Annex NA (informative) Bibliography DIN ISO 10381-1, Soil quality Sampling Part 1: Guidance on the design of sam
11、pling programmes DIN ISO 10381-6, Soil quality Sampling Part 6: Guidance on 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, So
12、il quality Determination of organic and total carbon after dry combustion (elementary analysis) DIN ISO 11268-3, Soil quality Effects of pollutants on earthworms Part 3: Guidance of the determination of effects in field situations DIN ISO 11272, Soil quality Determination of dry bulk density DIN ISO
13、 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 and sedimentation DIN ISO 11461, Soil quality Determination of soil water content as a volum
14、e 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 23611-4:2011-09 4 This page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 23611-4 July 2011 I
15、CS 13.080.30; 13.080.05 English Version Soil quality - Sampling of soil invertebrates - Part 4: Sampling, extraction and identification of soil-inhabiting nematodes Qualit du sol - Prlvement des invertbrs du sol - Bodenbeschaffenheit - Probenahme von Wirbellosen im Boden - Teil 4: Probenahme, Extrak
16、tion und Bestimmung von Boden bewohnenden Nematoden (ISO 23611-4:2007)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 for giving this European Standard the status of a national standa
17、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
18、 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
19、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
20、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 23611-4:2011: E(ISO 23611-4:2007) Partie 4: Prlvement, extraction et identif
21、ication des nmatodes du sol (ISO 23611-4:2007) Contents Page Foreword. 3 Introduction 4 1 Scope . 5 2 Terms and definitions. 5 3 Principle. 6 4 Reagents 7 5 Apparatus 7 5.1 Sampling 8 5.2 Extraction 8 5.3 Counting 8 5.4 Fixation and preparation of mass slides 9 5.5 Identification 9 6 Procedure 9 6.1
22、 General. 9 6.2 Sampling 9 6.3 Extraction 10 6.4 Counting 11 6.5 Fixation and preparation of mass slides 11 6.6 Identification 12 7 Data assessment. 12 8 Study report. 13 Annex A (informative) Figures of equipment and methods for nematological research 14 Annex B (informative) Information about the
23、availability of the Oostenbrink elutriator. 17 Annex C (informative) Information about the Baermann funnel/tray extraction method . 19 Bibliography . 2DIN EN ISO 23611-4:2011-09 EN ISO 23611-4:2011 (E) 21Foreword The text of ISO 23611-4:2007 has been prepared by Technical Committee ISO/TC 190 “Soil
24、quality” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 23611-4:2011 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 b
25、y 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 elements of this document may be the subject of patent rights. CEN and/or C
26、ENELEC 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 European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Rep
27、ublic, 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 the United Kingdom. Endorsement notice The text of ISO 23611-4:2007 ha
28、s been approved by CEN as a EN ISO 23611-4:2011 without any modification. 3DIN EN ISO 23611-4:2011-09 EN ISO 23611-4:2011 (E) Introduction This part of ISO 23611 has been drawn up since there is a growing need for the standardization of terrestrial zoological field methods. Such methods, mainly cove
29、ring the sampling, extraction and handling of soil invertebrates, are necessary for the following purposes: g127 biological classification of soils including soil quality assessment 15,17,28; g127 terrestrial bio-indication and long-term monitoring 9,10,13,24; g127 evaluation of the effects of chemi
30、cals 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 standardized methods is one of the most important reasons why bio
31、-classification and bio-assessment in terrestrial (i.e. soil) habitats has so far been relatively rarely used in comparison to aquatic sites. Nematodes are an important and major part of the soil fauna. Some authors estimate that this group is probably the most dominant one of the multicellular orga
32、nisms (Metazoa) on earth. Nematodes occur from the Antarctic to the tropics and from deep sea sediments to mountain regions. They are active in every place with sufficient water and organic material. The species diversity and functional variety are impressive. Nematodes are commonly known as parasit
33、es of animals and plants, but the major part of the nematode fauna participates in decomposition processes by feeding on bacteria and fungi. Nematodes occur in high numbers (5 000 to 100 000)/kg fresh soil and with a high (20 to 100) species diversity in almost every soil sample. Moreover, there is
34、a broad ecological spectrum of feeding types and food web relations among the nematodes such as bacterivores, fungivores, herbivores, predators and omnivores 27,28. These factors make the group highly suitable as indicators for ecological soil quality, but standardization of methods is urgently need
35、ed for comparison and combination of results. In the past 100 years, nematology has developed strongly from the viewpoint of agriculture, advisory sampling and phytosanitary regulations because some terrestrial nematodes cause a lot of damage in crops. With respect to methods, there are several “sch
36、ools” in different parts of the world with their own history, practical advantages and disadvantages. A comprehensive overview is given by Oostenbrink 14and Southey 22,23. The more recently described methods (or variants) are often developed with special interest to certain plant-parasitic species.
37、Since Bongers 4introduced the Maturity Index, the use of nematodes in bio-indication for soil quality has increased rapidly. Nematodes are now used for ecological soil research and monitoring in several countries all over the world. Monitoring activities make special demands on methodology, for inst
38、ance, that a large number of soil samples is processed on a routine basis against reasonable costs. Some of the methods originally developed for advisory sampling in agriculture are very suitable for ecological research. They form the basis for specific variants described in this document. 4DIN EN I
39、SO 23611-4:2011-09 EN ISO 23611-4:2011 (E) 1 Scope This part of ISO 23611 specifies a method for sampling and handling free-living nematodes from terrestrial field soils as a prerequisite for using them as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms). This part of
40、 ISO 23611 applies to all terrestrial biotopes in which nematodes occur. The sampling design of field studies in general is specified in ISO 10381-1. This part of ISO 23611 is not applicable to aquatic nematodes because these nematodes do not pass through the filter. Methods for some other soil orga
41、nism groups such as earthworms, enchytraeids or collembolans are covered in other parts of ISO 23611. The nematodes that are characterized by the proposed procedure are all the free-living forms of nematodes found in soil. They include non-plant-feeding nematodes as well as ectoparasitic plant-feedi
42、ng nematodes and free-living stage of endoparasitic nematodes. The quantification of obligate plant-feeding nematodes in roots requires specific methods. NOTE Basic information on the ecology of nematodes and their use as bio-indicators can be found in the bibliography. This part of ISO 23611 does n
43、ot cover the pedological characterization of the site 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 w
44、ater-holding capacity. 2 Terms and definitions For the purposes of this document, the following terms and definitions apply. 2.1 nematode small, non-segmented free-living worm (up to a few millimetres in length) belonging to the class Nematoda NOTE Nematodes without a soil-inhabiting stage are not i
45、ncluded in this context. 2.2 location study area or plot that is characterized based on the composition of (among others) the nematode fauna 5DIN EN ISO 23611-4:2011-09 EN ISO 23611-4:2011 (E) 2.3 bulk-sample composite soil sample made out of many small soil cores to get an impression of the average
46、 nematode composition 2.4 soil sampler tool to collect soil material in a quick and standardized way 2.5 Oostenbrink elutriator 1)metal funnel with an upward water flow to separate nematodes from larger soil particles See Figure A.3. 2.6 mass slide microscopic slide on which 300 to 400 nematodes are
47、 mounted for species identification 2.7 identification determination of the species, genus or family of an individual based on morphological characteristics (mouth parts, sexual organs, body ratios) with an identification key 2.8 colonizer persister (cp) scale ecological classification of nematodes,
48、 proposed by Bongers 4,5NOTE The principle is analogous to the r-K life strategies during succession, distinguished in fundamental ecology. Non-plant-feeding nematode families are classified to one of the five cp-groups. This is also the basis for the calculation of the Maturity Index. 3 Principle N
49、ematodes are collected in soil samples with a small cylindrical core (diameter: circa 2 cm; length: 10 cm) or an auger (see Figure A.2). For monitoring purposes, the soil samples are combined in a bulk-sample from a homogeneous area. The total number of samples to be taken depends on the investigated surface area and its homogeneity (e.g. pedology, culture). The individual samples can be gathered in the field in a standard plastic bag or plastic bucket. The combin
