ISO 17155:2002

Soil quality — Determination of abundance and activity of soil microflora using respiration curves

ISO 17155:2002 specifies a test method for determining the activity of the active aerobic, heterotrophic microbial biomass in soils. This method is applicable to the monitoring of soil quality and to the evaluation of the ecotoxic potential of soils and soil materials. It is also applicable to soils that are contaminated experimentally in the field or in the laboratory (chemical testing) and for soils sampled along contamination gradients in the field.

Qualité du sol — Détermination de l'abondance et de l'activité de la microflore du sol à l'aide de courbes de respiration

General Information

• Status: Withdrawn
• Publication Date: 07-Nov-2002
• Withdrawal Date: 07-Nov-2002
• ICS: 13.080.30 - Biological properties of soils
• Technical Committee: ISO/TC 190/SC 4 - Biological characterization
• Drafting Committee: ISO/TC 190/SC 4/WG 4 - Effects on soil micro-organisms
• Current Stage: 9599 - Withdrawal of International Standard
• Completion Date: 24-Sep-2012

Relations

• Revised: ISO 17155:2012 - Soil quality — Determination of abundance and activity of soil microflora using respiration curves
• Effective Date: 10-Jan-2009

Standards Content (Sample)

INTERNATIONAL ISO
STANDARD 17155
First edition
2002-11-01
Soil quality — Determination of abundance
and activity of soil microflora using
respiration curves
Qualité du sol — Détermination de l'abondance et de l'activité de la
microflore du sol à l'aide de courbes de respiration

Reference number
ISO 17155:2002(E)
© ISO 2002

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ISO 17155:2002(E)
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ISO 17155:2002(E)
Contents Page
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 3
5 Reagents . 3
6 Apparatus . 3
7 Sampling . 4
8 Procedure . 4
9 Calculation . 5
10 Test report . 7
Annex
A Results of an interlaboratory test carried out in Germany . 8
Bibliography. 11
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ISO 17155:2002(E)
Foreword
ISO (the International 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 body interested in a subject for which a technical committee has been established 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 closely with the International Electrotechnical
Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 3.
Draft International Standards adopted by the technical committees are circulated to the member bodies 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 International Standard may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights.
International Standard ISO 17155 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee
SC 4, Biological methods.
Annex A of this International Standard is for information only.
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INTERNATIONAL STANDARD ISO 17155:2002(E)
Soil quality — Determination of abundance and activity of soil
microflora using respiration curves
1 Scope
This International Standard specifies a test method for determining the activity of the active aerobic, heterotrophic
microbial biomass in soils. This method is applicable to the monitoring of soil quality and to the evaluation of the
ecotoxic potential of soils and soil materials. It is also applicable to soils that are contaminated experimentally in the
field or in the laboratory (chemical testing) and for soils sampled along contamination gradients in the field.
2 Normative references
The following normative documents contain provisions which, through reference in this text, constitute provisions of
this International Standard. For dated references, subsequent amendments to, or revisions of, any of these
publications do not apply. However, parties to agreements based on this International Standard are encouraged to
investigate the possibility of applying the most recent editions of the normative documents indicated below. For
undated references, the latest edition of the normative document referred to applies. Members of ISO and IEC
maintain registers of currently valid International Standards.
ISO 10381-6, Soil quality — Sampling — Part 6: Guidance on the collection, handling and storage of soil for the
assessment of aerobic microbial processes in the laboratory
ISO 10390, Soil quality — Determination of pH
ISO 10694, Soil quality — Determination of organic and total carbon after dry combustion (elementary analysis)
ISO 11277, Soil quality — Determination of particle size distribution in mineral soil material — Method by sieving and
sedimentation
ISO 11465, Soil quality — Determination of dry matter and water content on a mass basis — Gravimetric method
ISO 14238:1997, Soil quality — Biological methods — Determination of nitrogen mineralization and nitrification in
soils and the influence of chemicals on these processes
3 Terms and definitions
For the purposes of this International Standard, the following terms and definitions apply.
3.1
basal respiration rate
R
B
constant mass of CO released or mass of O consumed per unit mass of soil per unit time without substrate addition
2 2
See Figure 1.
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ISO 17155:2002(E)
3.2
substrate-induced respiration
R
S
constant mass of CO released or mass of O consumed per unit mass of soil per unit time shortly after addition of a
2 2
carbon substrate
See Figure 1.
NOTE If glucose is used as a carbon substrate, microbial biomass can be determined from the substrate-induced respiration rate
(see Reference [1] in the Bibliography).
3.3
lag time
t
lag
time from the addition of a growth substrate until exponential growth starts
See Figure 1.
NOTE It reflects the vitality of the growing organisms (see Reference [2] in the Bibliography).
3.4
growth rate
µ
rate constant during exponential increase of the respiration rate
See Figure 1.
3.5
respiratory activation quotient
Q
R
basal respiration rate divided by substrate-induced respiration rate
Q =R /R
R B S
3.6
time to the peak maximum
t
peakmax
time from addition of growth substrate to the maximum respiration rate
See Figure 1.
NOTE It reflects also the viability of the growing organisms.
3.7
cumulative CO evolution or O consumption
2 2
C
R
total area bounded by the soil respiration rate curve to the time axis from time of the addition of substrate to the time
See Figure 1.
3.8
soil material
material composed of excavated soil, dredged materials, manufactured soils, treated soils and fill materials
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ISO 17155:2002(E)
a
Addition of substrate
Figure 1 — Soil respiration rate before and after addition of an easily degraded substrate
4 Principle
The CO production or O consumption (respiration rate) from unamended soils as well as the decomposition of an
2 2
easily degraded substrate (glucose + ammonium + phosphate) is monitored regularly (at least every hour). Using the
CO production or O consumption data, the different microbial parameters (basal respiration, substrate-induced
2 2
respiration, lag time, respiratory activation quotient, t , C ) can be calculated.
peakmax R
5Reagents
5.1 Glucose, C H O .
6 12 6
5.2 Potassium dihydrogen phosphate, KH PO .
2 4
5.3 Diammonium sulfate, (NH ) SO .
4 2 4
5.4 Substrate, consisting of a mixture of 80 g of glucose (5.1), 13 g of diammonium sulfate (5.3) and 2 g of
KH PO (5.2) which is thoroughly ground and mixed in a mortar.
2 4
6Apparatus
Ordinary laboratory equipment and
6.1 Respirometer for continuous measurement of CO evolution or O consumption, maintained at a
2 2
◦ [3]
constant temperature (preferably 20 C). Suitable examples of equipment are given in ISO 16072 .
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ISO 17155:2002(E)
7 Sampling
7.1 Sample quantities
Choose the size of the soil samples size with respect to the apparatus (6.1) used and on the organic matter content
of the sample (7.1). Samples of organic horizons (e.g. mor layers) should not exceed 1g of organic matter (see
References [4] and [5]) in order to provide an optimal substrate/soil ratio (see 8.1). It is recommended to measure at
least three replicates per sample.
7.2 Sampling and storage
The recommendations in ISO 10381-6 for the collection, handling and storage of soil samples shall be followed.
7.3 Soil sample characteristics
Soil samples generating soil respiration curves can be obtained from mineral soils, organic soils, polluted and
unpolluted soils. Determine the following characteristics for each soil sample:
— particle size distribution in accordance with ISO 11277;
— water content in accordance with ISO 11465;
— water-holding capacity in accordance with annex A of ISO 14238:1997;
— pH in accordance with ISO 10390;
— organic matter content in accordance with ISO 10694.
8 Procedure
8.1 Test
Pre-incubate moist soil samples (preferably 40 % to 60 % of maximum water holding capacity or 0,01 MPa to
◦
0,03 MPa suction pressure) at 20C3 for d to 4d before the beginning of the measurement. Measure the basal
respiration of the sub-samples first. Measure the respiration rates until constant rates are obtained.
After measuring the basal respiration, add 0,2 g of the substrate (5.4) per gram of organic matter and mix
homogeneously with a spatula into the soil samples. If the organic matter content is less than 5% the substrate-to-
soil ratio is 1% [i.e. 1g of substrate added per 100g of soil (dry mass)].
Continue to measure the CO evolution or O consumption until the respiration rates decline (see Figure 1).
2 2
8.2 Influence of chemicals
The effect of chemicals on the soil microbial activity can be determined as follows. Using a range-finding test,
determine the concentration range that chemicals would likely have an effect on this activity. Test a single,
microbiologically active soil at five concentrations in a logarithmic series including a blank control in triplicate (for
example 0, 1, 3,2, 10, 32, and 100 times the lowest concentration). Use the test procedure specified in 8.1. Using this
simple test design, dose-response relationships can be established.
Before the start of the test, the test chemical may be added to the soil in one of the following manners:
— in an aqueous solution (depending on its solubility in water);
— in an organic solution using a water-miscible solvent (depending on the solubility in the solvent);
— mixed with a solid, e.g. coated on quartz sand (prior to mixing with the soil).
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ISO 17155:2002(E)
If the test chemical is added in the form of an organic solution, keep the amount of water-miscible solvent to the
minimum (< 1%) necessary for the application of the compound. Furthermore, take into account the possible
toxicity and biodegradability of the solvent used.
NOTE Long-term effects of chemicals can be detected by using different incubation times (weeks or months).
9Calculation
9.1 Microbial parameters
9.1.1 Basal respiration
Calculate the basal respiration (R ) as the average of the hourly respiration rate during a stable period.
B
9.1.2 Substrate-induced respiration
Calculate the substrate-induced respiration (R ) as the average of the values shortly after the substrate addition
S
when the respiration is fairly constant after substrate addition. A minimum o
...