1、BRITISH STANDARD 23611-4:2011Soil quality Sampling of soil invertebrates Part 4: Sampling, extraction and identification of soil-inhabiting nematodes ICS 13.080.05; 13.080.30nullnull nullnullnullnullnullnullnull nullnullnullnullnullnullnull nullnullnull nullnullnullnullnullnullnullnullnullnull nulln
2、ullnullnullnullnull nullnull nullnullnullnullnullnullnullnullnull nullnull nullnullnullnullnullnullnullnullnull nullnullnullBS EN ISONational forewordThis British Standard is the UK implementation of EN ISO 23611-4:2011. It is identical to EN ISO 23611-4:2011. It supersedes BS ISO 23611-4:2007, whic
3、h is withdrawn.The UK participation in its preparation was entrusted to Technical Committee EH/4, Soil quality.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Us
4、ers are responsible for its correct application.Compliance with a British Standard cannot confer immunity from legal obligations.BS EN ISO 23611-4:2011This British Standard was published under the authority of the Standards Policy and Strategy Committee on 29 February 2008 BSI 2011Amendments/corrige
5、nda issued since publicationDate Comments 31 August 2011 This corrigendum renumbers BS ISO 23611-4:2007 as BS EN ISO 23611-4:2011ISBN 978 0 580 73165 5EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 23611-4 July 2011 ICS 13.080.30; 13.080.05 English Version Soil quality - Sampling of soil i
6、nvertebrates - Part 4: Sampling, extraction and identification of soil-inhabiting nematodes (ISO 23611-4:2007) Qualit du sol - Prlvement des invertbrs du sol - Partie 4: Prlvement, extraction et identification des nmatodes du sol (ISO 23611-4:2007) Bodenbeschaffenheit - Probenahme von Wirbellosen im
7、 Boden - Teil 4: Probenahme, Extraktion 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 Stand
8、ard 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 three official versions (English
9、, 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 bodies of Austria, Belgium, Bul
10、garia, 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 Kingdom. EUROPEAN COMMITTEE FO
11、R 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 23611-4:2011: EForeword The text of ISO 2
12、3611-4:2007 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 23611-4:2011 by Technical Committee CEN/TC 345 “Characterization of soils” the secretariat of which is held by NEN. This Europe
13、an 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 eleme
14、nts 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 Eur
15、opean 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
16、the United Kingdom. Endorsement notice The text of ISO 23611-4:2007 has been approved by CEN as a EN ISO 23611-4:2011 without any modification. ii BS EN ISO 23611-4:2011EN ISO 23611-4:2011 (E)iiiContents Page1Scope . 12Terms and definitions. 13Principle. 24Reagents35Apparatus 35.1 Sampling 45.2 Extr
17、action 45.3 Counting 45.4 Fixation and preparation of mass slides 55.5 Identification 56Procedure 56.1 General. 56.2 Sampling 56.3 Extraction 66.4 Counting 76.5 Fixation and preparation of mass slides 76.6 Identification 87Data assessment. 88Study report. 9Annex A (informative) Figures of equipment
18、and methods for nematological research 10Annex B (informative) Information about the availabilityof the Oostenbrink elutriator. 13Annex C (informative) Information about the Baermann funnel/tray extraction method . 15Bibliography. 17BS EN ISO 23611-4:2011EN ISO 23611-4:2011 (E)iv Foreword ISO (the I
19、nternational 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 bodyinterested in a subject for which a technical committee h
20、as beenestablished 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 closel ywith the International Electrotechnical Commission (IEC) on all matters of electrotechnical
21、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 Standardsadopted by the technical committees are circulated to the member bodies
22、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. ISO shall not be held responsible for identifying any or
23、all such patent rights. ISO 23611-4 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 invertebrates: Part 1: Hand-sorting and formalin extraction of ear
24、thworms 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 soil-inhabiting nematodesBS EN ISO 23611-4:2011EN ISO 23611-4:2011 (E)vIntroduction This part of ISO 23611 has
25、been drawn up since there is a growing need for the standardization of terrestrial zoological field methods. Such m ethods, mainly covering the sampling, extraction and handling of soilinvertebrates, are necessary for the following purposes: biological classification of soils including soil quality
26、assessment 15,17,28; terrestrial bio-indication and long-termmonitoring 9,10,13,24; 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 shou
27、ld be remediated or not). In fact, the lack of such standardized methods is one of the most important reasonswhy bio-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
28、 the soil fauna. Some authors estimate that this group is probably the most dominant one of the multicellular organisms (Metazoa) on earth. Nematodes occur from theAntarctic to the tropics and from deep sea sediments to mountain regions. They ar e active in every place with sufficient water and orga
29、nic material. The species diversity and functional variety are impressive. Nematodes are commonly known as parasites of animals and plants, but the major part of the nematode faunaparticipates in decomposition processes by feeding on bacteria and fungi. Nematodes occur in high numbers (5 000 to 100
30、000)/kg fresh soil and with a high (20 to 100) speciesdiversity in almost every soil sample. Moreover, there is a broad ecological spectrum of feeding types and food web relations among the nematodes such as bacterivores, fungivores, herbivores,predators andomnivores 27,28. These factors make the gr
31、oup highly suitable as indicators for ecological soil quality, but standardization of methods is urgently needed 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 becau
32、se some terrestrial nematodes cause a lot of damage in c rops. Withrespect to methods, there are several “schools” in different parts of the world with their own history, practical advantagesand disadvantages.Acomprehensive overview is given by Oostenbrink 14 and Southey 22,23. The more recentlydesc
33、ribed methods (or variants) are often developed with special interest to certain plant-parasitic species.Since Bongers 4 introduced the MaturityIndex, the use of nematodes in bio-indication for soil quality has increased rapidly. Nematodes are now used for ecological soil research and monitoring in
34、several countries all over the world. Monitoring activities make special demands on methodology, for instance, that a large numberof soil samples is processed on a routine basis against reasonable costs. Some of the methods originallydeveloped for advisory sampling in agriculture are very suitable f
35、or ecological research. They form the basisfor specific variants described in this document. BS EN ISO 23611-4:2011EN ISO 23611-4:2011 (E)blank1Soil quality Sampling of soil invertebrates Part 4: Sampling, extraction and identification of soil-inhabiting nematodes 1 Scope This part of ISO 23611 spec
36、ifies a method for sampling and handling free-living nematodes from terrestrialfield soils as a prerequisite for using them as bio-indicators (e.g. to assess the quality of a soil as a habitat fororganisms).This part of ISO 23611 applies to all terrestrial biotopes in which nematodes occur. The samp
37、ling design offield 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 organism groups such as earthworms, enchyt raeids or collembolans arecovered in other
38、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-feeding nematodes and free-living stage of endoparasitic nematodes. The quantification
39、of obligate plant-feeding nematodes in roots requires specific methods. NOTE Basic information on the ecology of nematodes and their use asbio-indicators can be found in thebibliography. This part of ISO 23611 does not cover the pedological characterizationof the site which is highlyrecommendable wh
40、en sampling soil invertebrates. ISO10390, ISO10694, ISO11272, 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-holdingcapacity.2 Terms and definitions For the purposes of this document, the follow
41、ing terms and definitions apply.2.1 nematodesmall, non-segmented free-living worm (up to a fewmillimetres in length) belonging to the classNematoda NOTE Nematodes without a soil-inhabiting stage are not included in this context. 2.2 locationstudy area or plot that is characterized based on the compo
42、sition of (among others) the nematode fauna BS EN ISO 23611-4:2011EN ISO 23611-4:2011 (E)2 2.3 bulk-sample composite soil sample made out of many small soil cores to get an impression of the average nematodecomposition 2.4 soil sampler tool to collect soil material in a quick and standardized way2.5
43、 Oostenbrink elutriator 1)metal funnel with an upward water flow to s eparate nematodes from larger soil particlesSee Figure A.3. 2.6 mass slide microscopic slide on which300 to 400 nematodes are mounted for species identification 2.7 identificationdetermination of the species, genus or familyof an
44、individual based on mor phological characteristics (mouthparts, sexual organs, body ratios)with an identification key2.8 colonizer persister (cp)scale ecological classification of nematodes, proposed by Bongers 4,5NOTE The principle is analogous to the r-K life strategies during succession, distingu
45、ished 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 ofthe Maturity Index. 3 Principle Nematodes are collected in soil samples with a small cylindrical core (diameter: circa 2 cm; length: 10 cm)oran a
46、uger (see Figure A.2).For monitoringpurposes, 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 f ield in
47、 a standard plastic bag or plastic bucket. The combined bulk-sample is too large for direct examination and therefore it ismixed and subsampled. In the field and during transport to the laboratory, the soil samples shall be protectedagainst strong fluctuations in temperature, water-lossand heavy mec
48、hanical disturbance. They can be stored for at most four weeks at 4 C. NOTE 1The sampling method described above is derived from “the Dutch Method” 23 for determining the infestation of a field with potato-cyst nematodes, and has been used for many years in several European countries. 1) Oostenbrink
49、 elutriator is the trade name of a product supplied by firm Eijkelkamp, Giesbeek, NL( http:/www.eijkelkamp.nl ). This information is given for the convenience of users of this International Standard and doesnot constitute an endorsement by ISO of the product named. Equivalent products may be used if they can be shown tolead to the same results. BS EN ISO 23611-4:2011EN ISO 23611-4:2011 (E)3The Oostenbrink funnel method is recommended for r outine extractions of soil samples, for instance in amonitoring network. The Oost
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