1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationSoil quality Sampling of soil invertebratesPart 5: Sampling and extraction of soil macro-invertebratesBS EN ISO 23611-5:2013BS EN ISO 23611-5:2013National forewordThis British St
2、andard is the UK implementation of EN ISO 23611-5:2013. It is identical to ISO 23611-5:2011. It supersedes BS ISO 23611-5:2011, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee EH/4, Soil quality.A list of organizations represented on this committee can
3、 be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2013. Published by BSI Standards Limited 2013ISBN 978 0 580 79335 6 ICS 13.080.05; 13
4、.080.30 Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 December 2011.Amendments/corrigenda issued since publicationDate Text affected30 June 2013 This corrig
5、endum renumbers BS ISO 23611-5:2011 as BS EN ISO 23611-5:2013BRITISH STANDARDEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 23611-5 February 2013 ICS 13.080.30; 13.080.05 English Version Soil quality - Sampling of soil invertebrates - Part 5: Sampling and extraction of soil macro-invertebr
6、ates (ISO 23611-5:2011) Qualit du sol - Prlvement des invertbrs du sol - Partie 5: Prlvement et extraction des macro-invertbrs du sol (ISO 23611-5:2011) Bodenbeschaffenheit - Probenahme von Wirbellosen im Boden - Teil 5: Probenahme und Extraktion von Makroinvertebraten (Growirbellosen) im Boden (ISO
7、 23611-5:2011) This European Standard was approved by CEN on 5 February 2013. 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 bibl
8、iographical 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 respon
9、sibility 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, Former Yugoslav Repu
10、blic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMA
11、LISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 23611-5:2013: EForeword The text of ISO 23611-5:2011 has been prepared by Technic
12、al Committee ISO/TC 190 “Soil quality” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 23611-5:2013 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
13、 a national standard, either by publication of an identical text or by endorsement, at the latest by August 2013, and conflicting national standards shall be withdrawn at the latest by August 2013. Attention is drawn to the possibility that some of the elements of this document may be the subject of
14、 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 European Standard: Austria, Belgium, Bulgaria
15、, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey
16、 and the United Kingdom. Endorsement notice The text of ISO 23611-5:2011 has been approved by CEN as EN ISO 23611-5:2013 without any modification. ii BS EN ISO 23611-5:2013 ISO 23611-5:2013 (E) ISO 2013 All rights reservedEN iiiIntroductionThis part of ISO 23611 was prepared in response to a need to
17、 standardize sampling and extraction methods for soil macro-invertebrates in several European (temperate) and tropical countries. These methods are needed for the following purposes: biological classification of soils, including soil quality assessment (e.g. References 21, 32 and 41); terrestrial bi
18、o-indication and long-term monitoring (e.g. References 71, 79, 80 and 81).Data collected using standardized methods can be evaluated more accurately as it allows more reliable comparison between sites (e.g. polluted vs non-polluted sites, changes in land-use practices).Soils of the world host an abu
19、ndance of highly diverse macro-invertebrate communities. Their biology and ecology have been widely studied. Soil invertebrates are irreplaceable actors of soil formation and conservation in natural ecosystems. Their relevance to the soil system comes from their abundance and diversity, and also fro
20、m their role in key biological processes. They are sensitive indicators of soil quality and recognized agents of its fertility (e.g. References 63 and 56). Among the wide diversity of species, adaptive strategies and size ranges represented, one specific group, also called “soil ecosystem engineers”
21、, includes large invertebrates that actually determine the activities of other smaller organisms through the mechanical activities they produce in soil (e.g. References 24 and 49).Soil macro-invertebrates span a wide range of ecological functions in soil: decomposition of organic matter, through the
22、ir own activity and by stimulating the soils microbiological activity (e.g. References 8, 10 and 40), predation that plays an important part in food webs (e.g. References 16, 55, 61, 64 and 68), soil aggregation by the production of organo-mineral structures (e.g. nests, galleries, casts) that can l
23、ast for days, months or years, and soil bioturbation (e.g. Reference 32), etc. These characteristics, coupled with in-depth taxonomic knowledge, has enabled their use as study organisms in several research programmes dealing with the impacts of forest practices (e.g. References 18, 40, 50, 62, 65 an
24、d 75) or crop management practices (e.g. References 15, 25, 31, 33, 34, 37, 42, 60 and 66). These features make them suitable organisms for use as bio-indicators of changes in soil quality, especially with respect to land-use practices and pollution (e.g. References 26, 39, 48, 52, 53, 59, 65 and 79
25、).The method proposed in this part of ISO 23611 covers the sampling of all soil macro-invertebrates. However, the sampling of earthworms is already covered in ISO 23611-1. This method is described in ISO 23611-1:2006, Annex C, as an alternative sampling method for earthworms.BS EN ISO 23611-5:2013 I
26、SO 23611-5:2011 (E)iv INTERNATIONAL STANDARD Soil quality Sampling of soil invertebrates Part 5:Sampling and extraction of soil macro-invertebrates1 ScopeThis part of ISO 23611 specifies a method for sampling, extracting and preserving macro-invertebrates from soils, including the litter zone. The p
27、roposed method is a prerequisite for using these animals as bio-indicators (e.g. to assess the quality of a soil as a habitat for organisms). The main premise of this method is rapid assessment (completing the sampling of a plot in one or two days with only basic equipment and a small number of fiel
28、d assistants) in order to be able to address all the taxonomic groups of soil macro-invertebrates at the same time and in the same place. The Tropical Soil Biology and Fertility (TSBF) method has evolved and some modifications have been introduced in order to use it in temperate regions.The sampling
29、 and extraction methods in this part of ISO 23611 are applicable to almost all types of soil, with the exception of soils in extreme climatic conditions (hard, frozen or flooded soils) and matrices other than soil, e.g. tree trunks, plants or lichens.A sampling design is specified in ISO 23611-6.NOT
30、E 1 The method specified in this part of ISO 23611 is based on guidelines developed under the Tropical Soil Biology and Fertility Program (TSBF method)7.NOTE 2 Basic information on the ecology of macro-invertebrates and their use can be found in the references listed in the Bibliography.2 Terms and
31、definitionsFor the purposes of this document, the following terms and definitions apply.2.1macro-invertebratessoil organism whose longest dimension is greater than 10 mmNOTE See Annex A for further details.EXAMPLE These include especially the following groups: Oligochaeta, Gastropoda, Chilopoda, Dip
32、lopoda, Isopoda, Arachnida, plus various insects: Coleoptera, Orthoptera, Hymenoptera, Hemiptera, Dermaptera, Lepidoptera (larvae) and Diptera (larvae).2.2blotted massmass of individuals after preservation in formalin or ethanol (when the substance used for preservation has been absorbed by the tiss
33、ues)3 PrincipleSoil macro-invertebrates are collected in the field using a metallic frame to delimit the soil surface of the sampling point. Macro-invertebrates present in litter and soil are picked up separately. In temperate regions, a reagent is used to extract macro-invertebrates from soil. The
34、sampling is completed by hand-sorting. Animals are preserved and transported to the laboratory for further identifications (e.g. References 11, 12, 13, 14, 17, 19, 20, 22, 23, 27, 28, 29, 30, 35 ,36, ,38, 45, 46, 47, 54, 57, 70, 72, 73, 76, 77, 78 and 84). Abundance values are usually recalculated r
35、elative to area (1 m2).1BS EN ISO 23611-5:2013 ISO 23611-5:201 (E)14 Reagents4.1 Ethanol, (70 % volume fraction).4.2 Formalin formaldehyde solution, 4 % (volume fraction).Both 70 % ethanol and 4 % formalin should be available for the preservation of specimens (4 % formalin is more suitable for taxa
36、with soft body parts, which can be transferred to ethanol after about 4 d fixation).4.3 Formalin, 0,2 % (volume fraction), prepared by diluting 25 ml of formalin (39 %) in 5 l of water, for soil macro-invertebrate extraction.5 ApparatusUse standard laboratory equipment and the following.5.1 Petri di
37、shes.5.2 Stereo-microscope.5.3 Plastic vials.5.4 Entomological forceps.5.5 Pencil, notebook, water-resistant marker, labels.5.6 Tape measures.5.7 Knife (cut glass).5.8 Spade.5.9 Plastic-weave produce sacks, for spreading on the ground.5.10 Precision balance.5.11 Large flat plastic trays (500 mm 400
38、mm 100 mm), for sorting the soil and litter.5.12 Trowel.5.13 Small plastic trays.5.14 Fine forceps (or entomological forceps), pipette, fine paint brushes.5.15 Sample vials, in various sizes with secure alcohol-tight caps.5.16 Indian-ink pen (waterproof).5.17 Stiff card for labels, ranging compass.5
39、.18 Large strong plastic bags (sealable).2 BS EN ISO 23611-5:2013 ISO 23611-5:2011 (E)5.19 Table and plastic chairs, for sorting.5.20 Cover, for protection from heavy rain.5.21 Polyvinyl gloves, to protect hands from formalin.5.22 Metallic frame, preferably 250 mm 250 mm.Sample frame (250 mm 250 mm
40、50 mm) made of stainless steel and with sharpened edges to delimit the sampling point where animals are sampled from the litter layer and soil.5.23 Watering can.5.24 Pair of scissors, to cut vegetation inside the frame.5.25 Field balances.6 Field procedure6.1 GeneralSampling should take place when a
41、ccessible biodiversity is thought to be largest. In temperate regions, it corresponds to spring or autumn, and in the tropics, it should take place towards the end of the rainy season.When sampling soil invertebrates, it is strongly recommended that the site be physico-chemically characterized. In p
42、articular, pH, particle size distribution, C/N ratio, organic carbon content and water-holding capacity should be measured using ISO 10390, ISO 10694, ISO 11274, ISO 11277, ISO 11461, ISO 11465.6.2 Collecting macro-invertebrates from the litter zoneAt each sampling point (= monolith) (previously def
43、ined according to sampling design rules), a litter sample is collected using a metallic frame (5.22). The metallic frame is pressed into the litter by hand. The litter inside the frame is removed and checked manually in the field using a large tray (5.11). Litter invertebrates are preserved in 4 % f
44、ormalin (4.2).6.3 Collecting macro-invertebrates from soil6.3.1 GeneralIn temperate countries, the extraction of soil macro-invertebrates is carried out in two steps (see 6.3.2.1 and 6.3.2.2), while in tropical countries only the second step shall be performed (see 6.3.3).6.3.2 Temperate regions6.3.
45、2.1 Formalin extractionThe soil surface delimited by the metallic frame (5.22) is sprayed with 0,2 % formalin (4.3) using a watering can (5.23). Two applications of 1,5 l of formalin are performed at intervals of about 10 min. Soil invertebrates coming up to the surface are collected and preserved i
46、n vials (5.3) containing formalin (4.2).6.3.2.2 Hand-sorting of “passive” macro-invertebratesAt the end of the formalin extraction, the metallic frame (5.22) is removed and the upper 150 mm of soil is excavated within the frame area (250 mm 250 mm). The excavated soil is placed in a plastic bag (5.1
47、8) that can be closed with a cover to prevent animals from escaping from the soil sample.3BS EN ISO 23611-5:2013 ISO 23611-5:2011 (E)Appropriate sub-samples of soil are taken from the container and spread on a large tray (5.11). Macro-invertebrates are collected and preserved in vials (5.3) with for
48、malin (4.2). When hand-sorting is finished, the excavated soil is replaced to avoid creating holes on the sampling site.6.3.3 Tropical regionsIn tropical countries, soil macro-invertebrates are sampled using a 250 mm 250 mm 300 mm deep soil monolith. The monolith is isolated by cutting with a spade
49、(5.8) a few centimetres outside the quadrate (metallic frame) and then digging a 20 mm wide by 300 mm deep trench around it. This facilitates cutting of the sample into horizontal strata and collecting animals escaping from the block.The delimited block is divided into three layers, 0 mm to 100 mm, 100 mm to 200 mm and 200 mm to 300 mm, and the soil and litter material is hand-sorted in trays (5.11). S
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