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!$tTZ“1814955www.din.deDDIN EN ISO 16072Soil quality Laboratory methods for determination of microbial soil respiration(ISO 16072:2002)English translation of DIN EN ISO 16072:2011-09Bodenbeschaffenheit Laborverfahren zur Bestimmung der mikrobiellen Bodenatmung (ISO 16072:2002)Englische
3、bersetzung von DIN EN ISO 16072:2011-09Qualit du sol Mthodes de laboratoire pour la dtermination de la respiration microbienne du sol(ISO 16072:2002)Traduction anglaise de DIN EN ISO 16072:2011-09SupersedesDIN ISO 16072:2005-06www.beuth.deDocument comprises pagesIn case of doubt, the German-language
4、 original shall be considered authoritative.2708.11 DIN EN ISO 16072:2011-09 2 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee ISO/TC 190 “Soil quality” in collaboration with Technical Committee CEN/TC 345 “Characterization of soils” (S
5、ecretariat: 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 119-01-02 AA Abfall- und Bodenuntersuchung. The DIN Standards correspondin
6、g to the International Standards referred to in this document are as follows: ISO 4796-1 DIN EN ISO 4796-1 ISO 4796-2 DIN EN ISO 4796-2 ISO 4796-3 DIN EN ISO 4796-3 ISO 10381-6:1993 DIN ISO 10381-6:2009-09 ISO 11274:1998 E DIN ISO 11274:2011-01 ISO 11465:1993 DIN ISO 11465:1996-12 ISO 14240-1:1997 D
7、IN EN ISO 14240-1:2011-09 ISO 17155 DIN ISO 17155 Amendments No amendments have been made to DIN ISO 16072:2005-06. Previous editions DIN 19737: 2001-04 DIN ISO 16072: 2005-06 DIN EN ISO 16072:2011-09 3 National Annex NA (informative) Bibliography DIN EN ISO 4796-1, Laboratory glassware Bottles Part
8、 1: Screw neck bottles DIN EN ISO 4796-2, Laboratory glassware Bottles Part 2: Conical neck bottles DIN EN ISO 4796-3, Laboratory glassware Bottles Part 3: Aspirator bottles DIN EN ISO 14240-1:2011-09, Soil quality Determination of soil microbial biomass Part 1: Substrate-induced respiration method
9、DIN ISO 10381-6:2009-09, 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 E DIN ISO 11274:2011-01, Soil quality Determination of the water retention
10、 characteristics Laboratory methods DIN ISO 11465:1996-12, Soil quality Determination of dry matter and water content on a mass basis Gravimetric method DIN ISO 17155, Soil quality Determination of abundance and activity of the soil microflora using respiration curves DIN EN ISO 16072:2011-09 4 This
11、 page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 16072 June 2011 ICS 13.080.30 English Version Soil quality - Laboratory methods for determination of microbial soil respiration (ISO 16072:2002) Qualit du sol - Mthodes de laboratoire pour la dtermination de la res
12、piration microbienne du sol (ISO 16072:2002) Bodenbeschaffenheit - Laborverfahren zur Bestimmung der mikrobiellen Bodenatmung (ISO 16072:2002) This European Standard was approved by CEN on 10 June 2011. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the con
13、ditions for 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 exis
14、ts in three 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 sta
15、ndards bodies 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
16、 United Kingdom. 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 160
17、72:2011: EContents EN ISO 16072:2011 (E) DIN EN ISO 16072:2011-09 2 Page Foreword3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Procedure .6 4.1 General conditions 6 4.2 Choice of the measuring system .7 5 Measuring systems7 5.1 Determination of O2consumption by st
18、atic incubation in a pressure-compensation system.7 5.2 Determination of CO2release by titration in a static system8 5.3 Coulometric determination of CO2release in a static system10 5.4 Determination of CO2release using an infrared gas analyser in a flow-through system11 5.5 Determination of CO2rele
19、ase using gas chromatography in a flow-through system and a static system 14 5.6 Determination of soil respiration by pressure measurement in a static system 19 Bibliography 23 Foreword The text of ISO 16072:2002 has been prepared by Technical Committee ISO/TC 190 “Soil quality” of the International
20、 Organization for Standardization (ISO) and has been taken over as EN ISO 16072:2011 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 t
21、ext or by endorsement, at the latest by December 2011, and conflicting national standards shall be withdrawn at the latest by December 2011. 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 resp
22、onsible 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 Republic, Denmark, Estonia, Finl
23、and, 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 16072:2002 has been approved by CEN as a EN
24、ISO 16072:2011 without any modification. EN ISO 16072:2011 (E) DIN EN ISO 16072:2011-09 3 Introduction This International Standard is derived from the German standard DIN 19737 (see 1). It describes methods for the determination of microbial soil respiration in the laboratory. Microbial soil respira
25、tion results from the mineralization of organic substances. In this process, organic substances are oxidized to the end products carbon dioxide and water, with concurrent uptake of O2for aerobic microorganisms. The soil respiration is measured by the determination of O2consumption and/or by CO2relea
26、se. Respiration is a measure of the overall activity of soil microorganisms. EN ISO 16072:2011 (E) DIN EN ISO 16072:2011-09 4 1 Scope This International Standard describes methods for the determination of soil microbial respiration of aerobic, unsaturated soils. The methods are suitable for the dete
27、rmination of O2uptake or CO2release, either after addition of a substrate (substrate-induced respiration), or without substrate addition (basal respiration). This International Standard is applicable to the measurement of soil respiration in order to: determine the microbial activity in soil (see 3)
28、; establish the effect of additives (nutrients, pollutants, soil improvers, etc.) on the metabolic performance of microorganisms; determine the microbial biomass (see 4); determine the metabolic quotient qCO2. 2 Normative references The following referenced documents are indispensable for the applic
29、ation of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 10381-6:1993, Soil quality Sampling Guidance on the collection, handling and storage of soil for the assessment
30、of aerobic microbial processes in the laboratory ISO 11274:1998, Soil quality Determination of the water-retention characteristic Laboratory methods ISO 11465:1993, Soil quality Determination of dry matter and water content on a mass basis Gravimetric method 3 Terms and definitions For the purposes
31、of this document, the following terms and definitions apply. 3.1 basal respiration microbial soil respiration without addition of nutrients 3.2 substrate-induced respiration SIR microbial soil respiration after addition of nutrients NOTE Glucose is an example of an added nutrient. EN ISO 16072:2011
32、(E) DIN EN ISO 16072:2011-09 5 3.3 microbial activity metabolic performance of microorganisms NOTE It can be measured, for example, as O2uptake or CO2release. 3.4 metabolic quotient qCO2specific metabolic activity of soil microorganisms, which can be calculated as the quotient basal respiration: mic
33、robial biomass NOTE Metabolic quotient is usually expressed as milligrams of CO2carbon released per hour per gram of microbial biomass carbon. 3.5 rate of CO2formation O2consumption RCO2 RO2 amount of CO2released O2consumed per time unit from a mass unit of soil NOTE 1 Soil respiration is usually me
34、asured as the rate of CO2formation or O2consumption. NOTE 2 It is usually expressed as milligrams CO2or O2 per gram per hour (mg CO2or O2g1h1). 3.6 microbial biomass mass of intact microbial cells in a given soil NOTE This is usually estimated from the measurement of carbon or nitrogen content of th
35、ese cells. 4 Procedure 4.1 General conditions 4.1.1 Soil sampling and storage Sample, store and pre-incubate soils in accordance with ISO 10381-6, independently of the choice of the procedure and the respiration parameter to be measured (basal respiration, SIR). 4.1.2 Measuring and incubation condit
36、ions Soil respiration is strongly influenced by water content and temperature. Therefore these parameters should be recorded in the final report. At suction pressures 0,03 MPa, the soil respiration will decrease considerably. The water content of the test soil is optimal when it corresponds with a p
37、ore water pressure of 0,01 MPa to 0,03 MPa (measured with an accuracy of 5 %, in accordance with ISO 11274) or 40 % to 60 % of the maximum water-holding capacity, respectively. A stable temperature should be used. Incubation temperatures between 20 C and 30 C are generally recommended, but other tem
38、peratures may be used if required. In the description of the methods, examples of incubation temperatures are given as well as the accuracy of temperature maintenance and measurement. If a method is used for the determination of soil microbial biomass, a temperature of 22 C is recommended because bi
39、omass calculations have been calibrated to this temperature. When soil samples are compared with respect to soil respiration, they should have the same moisture status (pore water pressure or percentage of maximum water-holding capacity). EN ISO 16072:2011 (E) DIN EN ISO 16072:2011-09 6 4.2 Choice o
40、f the measuring system Each measurement method has its own advantages and disadvantages. Care is needed, because the results obtained by O2uptake and by CO2release are not strictly compatible. It is the responsibility of the investigator to decide which of these methods is to be used. One of the sys
41、tems described in Clause 5 should be used. Systems for measuring CO2do not distinguish between CO2released from microbial activities and CO2resulting from abiotic processes. For alkaline soils and soils with a high organic matter content, which can release considerable amounts of abiotically release
42、d CO2, methods using O2uptake are recommended. NOTE The advantages and disadvantages are described in the individual descriptions of the methods. 5 Measuring systems 5.1 Determination of O2consumption by static incubation in a pressure-compensation system 5.1.1 Principle The determination is based o
43、n the measurement of O2consumption during incubation of a soil sample in a closed system. The O2in the system is replenished electrochemically. The CO2released is absorbed by calcium hydroxide Ca(OH)2. Key A reaction vessel 1 soil sample 4 electrolyte B oxygen generator 2 CO2absorbent 5 electrodes C
44、 pressure indicator 3 pressure cell 6 recorder with display Figure 1 Determination of O2consumption (showing connection of a measuring unit) 5.1.2 Apparatus A detailed description of the apparatus can be found in 4; the essential features are as follows. EN ISO 16072:2011 (E) DIN EN ISO 16072:2011-0
45、9 7 The measuring system (see Figure 1) consists of a water bath with temperature control containing measuring units each comprising a reaction vessel (A) in which a CO2absorption device (2) is suspended from the stopper, an O2generator (B) and a pressure indicator (C). The vessels (A, B, C) of the
46、measuring unit form together a closed system, connected to each other by tubing. In this way fluctuations in atmospheric pressure will not influence the results. The CO2released is absorbed by the calcium hydroxide (2). The consumption of O2due to respiration results in a negative pressure which act
47、ivates the pressure indicator (C). This drives the electrolytic O2formation as well as the display and graphical registration of measuring values on a recorder (6). The O2consumption is shown directly, in milligrams of oxygen (mg O2) on a digital display. The system can be obtained commercially1)and
48、 detailed instructions should be given in the suppliers manual. 5.1.3 Procedure Use 50 g to 100 g of field-moist, sieved (2 mm) soil for the measurements. The O2consumption should not be measured during the first 2 h, the time needed to reach equilibrium in the system. 5.2 Determination of CO2releas
49、e by titration in a static system 5.2.1 Principle The soil is incubated in a closed vessel and the released CO2is absorbed in a solution of sodium hydroxide. After back-titration of the non-consumed sodium hydroxide, the amount of CO2released is calculated. The method is suitable for large numbers of samples, and up to 80 respiration measurements per working day are possible. 5.2.2 Reagents 5.2.2.1 CO2-free water Boil distilled water and after cooling store it in flasks closed
