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1、 Reference number ISO 23611-2:2006(E) ISO 2006 INTERNATIONAL STANDARD ISO 23611-2 First edition 2006-02-01 Soil quality Sampling of soil invertebrates Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina) Qualit du sol Prlvement des invertbrs du sol Partie 2: Prlvement et extr
2、action des micro-arthropodes (Collembola et Acarina) Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 19:36:41 MDTNo reproduction or networking permitted without license from IHS
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6、ddress given below. ISO 2006 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or I
7、SOs member body in the country of the requester. ISO copyright office Case postale 56 CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2006 All rights reserved Copyright International Organization for Standardizati
8、on Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 19:36:41 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 23611-2:2006(E) ISO 2006 All rights reserved iii Contents Page Foreword iv Introduction v 1 Scope
9、 . 1 2 Terms and definitions. 1 3 Principle. 1 4 Test materials 2 4.1 Biological material 2 4.2 Reagents 2 5 Apparatus 3 6 Procedure 4 6.1 Collecting the soil samples . 4 6.2 Extracting Collembola and Acarina from soil samples 4 6.3 Sorting, preserving and identifying Collembola and Acarina 5 7 Asse
10、ssment of results 6 8 Study report. 6 Annex A (informative) Species determination in collembolans and mites 7 Annex B (informative) Alternative methods for sampling of micro-arthropods 9 Bibliography. 10 Copyright International Organization for Standardization Provided by IHS under license with ISO
11、Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 19:36:41 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 23611-2:2006(E) iv ISO 2006 All rights reserved Foreword ISO (the International Organization for Standardization) is a worldwide federation
12、 of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee.
13、International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance wit
14、h the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approv
15、al 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 all such patent rights. ISO 23611-2 was prepared by Technical Commit
16、tee 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 earthworms Part 2: Sampling and extraction of micro-arthropods (Collemb
17、ola and Acarina) Part 3: Sampling and soil extraction of enchytraeids Part 4: Sampling, extraction and identification of free-living stages of terrestrial nematodes Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/997
18、2545001 Not for Resale, 04/19/2007 19:36:41 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 23611-2:2006(E) ISO 2006 All rights reserved v Introduction This part of ISO 23611 has been drawn up since there is a growing need for the standardization of sampling and extract
19、ion methods of soil micro-arthropods. These methods are needed for the following purposes: biological classification of soils including soil quality assessment (e.g. References 31, 32, 35, 41, 45, 46); terrestrial bioindication and long-term monitoring (e.g. References 1, 7, 17, 40, 42). Data collec
20、ted 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 studied in soil ecology. Their relevanc
21、e 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 soil food webs 9. These characteri
22、stics, 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) or crop management practices (e.g.
23、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. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standard
24、s 1/9972545001 Not for Resale, 04/19/2007 19:36:41 MDTNo reproduction or networking permitted without license from IHS -,-,- Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 19:36
25、:41 MDTNo reproduction or networking permitted without license from IHS -,-,- INTERNATIONAL STANDARD ISO 23611-2:2006(E) ISO 2006 All rights reserved 1 Soil quality Sampling of soil invertebrates Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina) 1 Scope This part of ISO 23
26、611 specifies 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
27、 be found in the 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,
28、plants or lichens. 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 recommen
29、dable 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 water-holding capacity. 2 Terms and definitions For the purposes of this docume
30、nt, the following 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), sp
31、ringtails (Collembola), 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
32、tubes (or plastic 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
33、, abundance values can be recalculated related to area (usually 1 m2), volume or weight (usually 1 kg). Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 19:36:41 MDTNo reproductio
34、n or networking permitted without license from IHS -,-,- ISO 23611-2:2006(E) 2 ISO 2006 All rights reserved 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 loamy soils and a Kempson extracto
35、r 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 abdominal segments and three pre-geni
36、tal 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 tenaculum, that holds the jumping
37、 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 into 18 families 17. Soil mites a
38、re 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 (Oribatida), Mesostigmata (Gamasi
39、da), 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). 4.2.2 Formalin formaldehyde solu
40、tion 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 Trne fixative, used to preserve th
41、e 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 hydrogen chloride (5 ml). 4.2.10 L
42、actophenol 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 the microscope. 4.2.12 Ethanol, 70 %
43、 to 75% (volume fraction), used for fixation and preservation (in this case, also in combination with glycerine, 10:1). Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 19:36:41 M
44、DTNo reproduction or networking permitted without license from IHS -,-,- ISO 23611-2:2006(E) ISO 2006 All rights reserved 3 4.2.13 Hoyers medium, used to mount Collembola specimens composed of distilled water (50 ml), gum-arabic (30 g), chloral hydrate (200 g) and glycerine (20 ml). 5 Apparatus Use
45、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, wit
46、h excavated area in the centre, and lamellae. 5.9 Electrical heating plate. 5.10 Plastic vials. 5.11 Ceramic heating elements. 5.12 Pencil, notebook, water resistant marker, labels. 5.13 Split corer Sampling device made of stainless steel or aluminium (40 cm long and e.g. 5,6 cm diameter may be used
47、; the size and diameter should not differ considerably from these numbers in order to maintain comparable conditions), used to collect soil cores (samples). It can be composed of two independent parts that fit together along the corer main axis or it can consist of one tube. On the top, it has a han
48、dle and on the bottom, a cutting edge. 5.14 MacFadyen apparatus High-gradient (multiple) device used to extract micro-arthropods from soil samples. The principle is to create an artificial temperature gradient between the canister where the sample is placed (hot) and the collecting device below (cold), inducing a negative thermotactic (at the same time a positive hygrotactic, ne