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Method for spiking soil samples with organic compounds.

Ulla C Brinch¹, Flemming Ekelund, Carsten S Jacobsen

¹Department of Geochemistry, Geological Survey of Denmark and Greenland, Copenhagen.

Applied and Environmental Microbiology

|March 28, 2002

Summary

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Acetone and dichloromethane harm soil microbes when applied directly. A new protocol adding solvents to 25% of soil, followed by mixing, minimizes these adverse effects on soil microorganisms.

Area of Science:

Environmental Science
Microbiology
Soil Science

Background:

Organic solvents like acetone and dichloromethane are used to introduce polycyclic aromatic hydrocarbons (PAHs) into soil for experiments.
These solvents can negatively impact indigenous soil microorganisms, affecting experimental outcomes.
Understanding these solvent effects is crucial for accurate soil contamination studies.

Purpose of the Study:

To investigate the harmful side effects of acetone and dichloromethane on indigenous soil microorganisms.
To compare different contamination protocols for their impact on soil microbial communities.
To propose an optimized protocol for spiking soil samples that minimizes solvent-induced damage.

Main Methods:

Two organic solvents (acetone, dichloromethane) were applied to soil samples using three distinct protocols.

Related Experiment Videos

Protocols involved applying solvents to the whole soil sample, 25% of the soil volume, or 80% of the soil volume.

Indigenous protozoa and bacteria populations were monitored, along with the introduction of a tagged bacterium (Pseudomonas fluorescens VKI171 SJ132).

Main Results:

Direct application of both solvents to the entire soil sample caused severe damage to protozoa and bacteria.
Dichloromethane application to whole soil reduced protozoa to undetectable levels, with limited recovery in diversity.
Introduced P. fluorescens showed a 1,000-fold increase in growth, likely due to reduced predation by indigenous protozoa.

Conclusions:

Direct application of organic solvents to soil samples significantly harms indigenous microbial communities.
A proposed protocol involving solvent application to 25% of the soil, followed by evaporation and mixing, minimizes these detrimental effects.
This optimized method ensures more reliable experimental results when studying soil contamination with low-water-solubility compounds.