Traceability of

Ammonia-Oxidizing Bacteria
in Compost-Treated Soils
Traceability of
Ammonia-Oxidizing Bacteria
in Compost-Treated Soils
Gerd Innerebner
1
, Brigitte Knapp
1
, Martin Romantschuk
2
,
Tuija Vasara
2
, Heribert Insam
1
1
Dept. of Microbiology, University of Innsbruck, Innsbruck, Austria
2
Dept. of Ecological and Environmental Sciences, University of Helsinki, Lahti, Finland
AIMS:
Composts are increasingly used as environmentally safe biofertilizers in
sustainable agriculture all over the world. In this study, we investigated
if the continued application of different composts would, apart from
effects on the ecosystem level, leave a microbial imprint at the population
level of ammonia-oxidizing bacteria (AOB).
We applied molecular techniques such us PCR-DGGE, cloning, sequencing
and real-time PCR.
MATERIALS & METHODS
A long-term fertilization experiment was started in 1991 near
Linz, Austria, where the effect of different composts on soil
quality and crop yield is studied (Aichberger et al. 2000). The
field consists of randomized plots with the following 8
treatments (four replicates each):
RESULTS
GELCOMPAR cluster analysis of DGGE profiles differentiated
between composts (A), mineral-fertilized soils (B) and compost-
treated soils (C):
DISCUSSION
Results similar to our cluster analysis have been reported by
Ros et al. (2005), who used a general bacterial primer pair for
PCR-DGGE in the same set of soils. Our data suggest that the
different origin of the organic matter does not have a
significant effect on the AOB community and that the addition
of a certain microbial community inherent to the different
composts is irrelevant for the ammonia-oxidizer community in
soils. Since the AOB composition of the composts is not
reflected in the AOB community of the soils, the hypothesis of
a direct traceability of the compost community in soils must be
rejected. Quantitative data from real-time PCR confirmed
these findings.
REFERENCES
Ros M, Klammer S, Knapp B, Aichberger K and Insam H (2005) Long-term
effects of soil compost amendment on functional and structural diversity and
microbial activity. Submitted for publication.
Aichberger K, Wimmer J and Mayr R (2000) Auswirkung der
Kompostanwendung auf Ertrag und Bodeneigenschaften. In: 6. Alpenlndisches
Expertenforum, 16. - 17. Mrz pp. 81-83.
ACKNOWLEDGEMENTS
Thanks to AGES for maintaining the research plots and to the
staff members of the Dept. of Environmental and Ecological
Sciences in Lahti (Finland) for support with real-time PCR. This
study has been funded by the Austrian Science Fund (FWF).
0E+00
1E+07
2E+07
3E+07
4E+07
5E+07
6E+07
SOILS + N(min) SOILS + N(org)
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SOIL GRE BIO SEW MAN
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cluster 7
cluster 6
cluster 4
cluster 3
SAMPLING
DNA EXTRACTION
DGGE CLONING
CLUSTER ANALYSIS
PCR
MULTIPLE
ALIGNMENT
SEQUENCING REAL-TIME PCR
quantitative
approach
qualitative
approach
SAMPLING
DNA EXTRACTION
DGGE CLONING
CLUSTER ANALYSIS
PCR
MULTIPLE
ALIGNMENT
SEQUENCING REAL-TIME PCR
quantitative
approach
qualitative
approach
After soil sampling in October 2003, DNA was extracted and
nested PCR was applied to amplify 16S rDNA fragments
specific for ammonia-oxidizing bacteria (AOB). The community
structure of AOB in soils and composts was analyzed by DGGE.
To carry out a phylogenetic analysis, DNA fragments were
cloned and sequenced.
Quantitative real-time PCR was performed to determine the
abundance of AOB.
Sequencing of screened clones revealed a characteristic AOB
community structure for each investigated soil and compost
(Fig. 1). All AOB-like sequences grouped within the Nitrosospira
cluster 3 and 4 and within the Nitrosomonas cluster 6 and 7.
The average AOB abundance in compost-treated soils was two
times higher than in mineral-fertilized soils (4.310
7
and
1.910
7
, respectively; one-way ANOVA , p0.05, Fig. 2). The
AOB population size in the composts varied considerably,
however, it was directly related to their total N content (data
not shown).
Fig. 1 Fig. 2
Gradient gel
Research plots
Sewage
sludge
(SEW)
Manure
(MAN)
Green
waste
(GRE)
Organic
waste
(BIO)
Composts
120 kg 80 kg 40 kg 0 (control) Mineral
fertilizer
Treatment
Sewage
sludge
(SEW)
Manure
(MAN)
Green
waste
(GRE)
Organic
waste
(BIO)
Composts
120 kg 80 kg 40 kg 0 (control) Mineral
fertilizer
Treatment
CONCLUSIONS:
Compared to control and mineral fertilization,
long-term compost amendment left an imprint on
the level of ammonia oxidizers. Differences
among composts were not detectable, thus an
indirect effect is suggested, rather than a direct
effect of the AOB community in the composts.
BAGECO 8 - Lyon, 26 - 29 June 2005