european journal of soil biology 44 (2008) 231–237

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Original article

Soil microbiological and biochemical properties

for assessing the effect of agricultural management
practices in Estonian cultivated soils

Marika Truua,*, Jaak Truua, Mari Ivaskb

a
Faculty of Biology and Geography, University of Tartu, 23 Riia Street, 51010 Tartu, Estonia
b
Tartu College of Tallinn University of Technology, 78 Puiestee Street, 51008 Tartu, Estonia

article info abstract

Article history: A set of soil microbiological and biochemical properties was used to assess the influence of
Received 3 September 2007 agricultural practices such as rotation, usage of pesticides, and fertilizers on the three most
Accepted 19 December 2007 widespread soil types (Calcaric Regosols, Calcaric Cambisols and Stagnic Luvisols) in the
Published online 18 June 2008 fields of horticultural farms throughout Estonia. Microbial biomass, dehydrogenase and
alkaline phosphatase activity were significantly higher in Calcaric Regosols, whereas mea-
Keywords: sured soil chemical parameters showed practically no difference among soil types. Multi-
Agricultural management practice variate exploratory analysis of soil biochemical and microbiological parameters clearly
Microbial activity distinguished soils with different management practices when the effect of soil type was
Microbial biomass taken into account in data analysis. Activity of dehydrogenase, potential nitrification, N-
Soil type mineralisation, and microbial biomass contributed most strongly to the differentiation of
soils from differently managed fields. Soils managed according to organic farming princi-
ples were generally characterized by elevated microbiological parameter values, but at
the same time the variation of those parameters among soils from these fields was also
highest. The application of organic manure positively affected microbial biomass, N-
mineralisation, potential nitrification, dehydrogenase and acidic phosphatase activity.
Data analysis indicated that the amount of mineral nitrogen fertilizers added over time
has a stronger effect on microbial biomass than the amount added in a given year.
Legume-based crop rotation increased soil respiration and microbial biomass.
ª 2008 Elsevier Masson SAS. All rights reserved.

1. Introduction fertilizers and agrochemicals followed. At the beginning of

There have been two major changes in Estonian agricultural
management practice during the last century. At the begin-
ning of the 1940s land was taken from private owners and
large collective farms were established. Fifty years of very
intensive agricultural practice with high inputs of mineral
the 1990s, the land was returned to private owners, but during
the last decade the economic situation and agricultural policy
have been unfavorable for agricultural land users. Since 1989
cropping area has steadily decreased by about half (from
975,000 ha in 1989 to 461,000 ha in 2003). The first organically
managed farms were established at the very end of the

* Corresponding author. Tel.: þ372 7375 014; fax: þ372 7420 286.
E-mail address: marika.truu@ut.ee (M. Truu).
1164-5563/$ – see front matter ª 2008 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejsobi.2007.12.003
232 european journal of soil biology 44 (2008) 231–237

1990s. Nowadays ecologically managed land comprises about
5% of all agricultural land and only 14% of this land is used for
field crops. However, there are also still a number of farmers
who do not strictly follow any particular (organic or conven-
tional) management practice.
Soil is a complex environment, where microorganisms
play a crucial role in nutrient cycling and the degradation of
different pollutants (herbicides, pesticides, PAH-s, phenols,
etc.) contributing in this way to the maintenance of soil qual-
ity [8,14,39]. On the other hand, microbial activities are
strongly dependent on nutritional and other chemical and
physical conditions of the soil and respond rapidly to changes
in soil properties. Microorganisms are considered sensible
indicators when monitoring changes in soil status affected
by agricultural management, but the meaningful set of micro-
biological indicators still remains an object of debate
[3,11,31,35].
Microbial biomass is considered to be an integrative, bio-
logically meaningful, management sensitive, and measurable
signal in the soil [30]. Its turnover rate is much faster than that
of total soil organic matter, and based on the dynamics of soil
microbial biomass content, longer-term trends in soil total
organic matter content can be predicted [34]. The influence
of soil management on the organic matter C and N turnover
capacity of microbial biomass has been pointed out in many
studies focussing on microbial biomass and activity measure-
ments in arable soils [4,10,28]. Soil respiration is considered to
be one of the well-established parameters for monitoring
decomposition, but it is highly variable and can fluctuate
widely depending on substrate availability, moisture content
and temperature [34]. N-mineralisation reflects the quality
and quantity of soil organic nitrogen and links the substrate
with the functioning and activity of a range of soil organisms.
In order to estimate the effects of soil management, land use
and specific conditions on soil microbial activity, short-term
laboratory measurements, including enzymatic activities are
used [3,12]. The advantage of standardizing environmental
factors is that it allows the comparison of soils of different or-
igins, but the results obtained represent the potential activity
only and must be interpreted with reservations [31].
The objective of this work was to use a set of soil microbi-
ological and biochemical indicators to evaluate the effect of
different management practices in three Estonian soil types.
2. Materials and methods
2.1. Soil type and management description
Twenty-three study areas with three most widespread soil
types (Calcaric Regosols, Calcaric Cambisols and Stagnic Luvi-
sols) throughout Estonia were selected (Table 1). On Calcaric
Cambisols and Stagnic Luvisols eight fields, and on Calcaric
Regosols seven fields with different management practices
were selected. Three years history of agricultural practices
(cover crops, amounts of mineral and organic fertilizers and
different kinds of pesticides used) was recorded. Mineral
nitrogen was applied on 17 fields and the amounts ranged

Table 1 – Soil and site characteristics of studied fields

Site no. Location Soil typea Mineral nitrogen, Organic fertilizers, Pesticidesc Legumes/rape
sum of three sum of three
years (kg ha1) yearsb (t ha1)

1 58 160 0800 N 22 030 0200 E CR No/no

2 58 190 0800 N 22 000 4500 E CR 120 Yes/no
3 58 370 1300 N 26 310 1500 E CR 60 65 H Yes/no
4 58 580 4500 N 24 430 0100 E CR 70 GM H Yes/no
5 58 580 4700 N 24 420 2500 E CR 100 H, F No/yes
6 58 580 5400 N 24 420 4200 E CR 50 H Yes/no
7 58 390 2700 N 26 370 4000 E CC 70 GM Yes/no
8 58 430 0300 N 26 390 2800 E CC 315 H, I Yes/no
9 59 090 2700 N 25 450 1900 E CC 248 H, I, F No/yes
10 59 100 5100 N 25 460 2100 E CC 249 H, I, F No/yes
11 59 100 4200 N 25 460 0900 E CC 175 40 H No/no
12 58 580 5800 N 24 420 5300 E CC 70 GM H, F Yes/no
13 58 580 5700 N 24 430 0000 E CC 140 H Yes/no
14 58 580 5200 N 24 430 1000 E CC 146 H Yes/no
15 58 410 3500 N 26 340 0300 E SL GM Yes/no
16 58 410 4800 N 26 330 4900 E SL GM Yes/no
17 58 370 0300 N 26 300 5900 E SL 25 Yes/no
18 58 330 1000 N 25 330 4400 E SL 285 H, I, F No/yes
19 58 330 0300 N 25 340 0200 E SL 309 H, I, F No/yes
20 58 330 4300 N 25 330 2300 E SL 245 H, I No/no
21 58 420 0300 N 26 350 3800 E SL 100 40 H, I No/yes
22 58 260 2500 N 22 010 0800 E CR 80 No/yes
23 58 420 3000 N 26 390 2200 E SL 182 H No/yes

a CR – Calcaric Regosols, CC – Calcaric Cambisols, SL – Stagnic Luvisols.

b GM – green manure, numbers are shown for brown manure.
c H – Herbicides, I – Insecticidas, F – Fungicides.
 european journal of soil biology 44 (2008) 231–237
from 15 kg ha1 per year (in three cases) to 142.5 kg ha1 per
year (in one case during the last three years). Animal manure
was applied on seven fields and six fields have received green
manure. The amount of brown manure applied per year
ranged from 25 t ha1 to 40 t ha1. Three of the studied fields
had received both organic and mineral fertilizers simulta-
neously. Seven fields had oilseed rape in rotation (one or
two years during the last three years) and were also treated
with different pesticides – herbicides on all seven fields, insec-
ticides and fungicides on six fields. All together 16 fields were
treated with herbicides during the last three years. In 11 fields
at least once during the three years leguminous crops (mainly
clover) were grown. One field had not been treated with any
fertilizers or pesticides during the last three years, nor did it
have any leguminous crops in rotation.
2.2. Soil sampling
The samples were taken from the fields with a soil corer
(B2 cm) from 0 to 20 cm layer at the end of October 2003 and
composite samples were made for each field. For the compos-
ite samples (one to three composite samples depending on the
size of the field, one composite sample per 2 ha) 25 subsam-
ples were randomly collected from each field. The fresh soil
samples were sieved (<2 mm) and stored at 4  C until the anal-
yses were carried out. From the sieved samples, soil organic
matter content (the loss on ignition method), dry matter
content, pHKCl, total nitrogen, total potassium, and available
phosphorus content were determined and microbiological
analyses were performed.
2.3. Microbiological biomass and activities
Substrate-induced respiration (SIR) by Isermeyer technique
was applied to measure metabolically active microbial
biomass carbon. Glucose (0.4 g 100 g1 soil) was added to 20 g
of field moist soil and then incubated in a closed vessel for
4 h at 22  C in the dark. The CO2 produced was absorbed in
0.1 M sodium hydroxide and quantified by titration. The mi-
crobial biomass C was calculated according to Beck et al. [2].
Soil microbial respiration rate (basal respiration) was mea-
sured by titration according to Öhrlinger [41]. Twenty gram
of soil was incubated in a closed vessel for 24 h at 25  C. The
CO2 produced was absorbed in 0.05 M sodium hydroxide,
quantified by titration, and the respiration rate was calcu-
lated. The microbial metabolic quotient qCO2 was calculated
as the ratio between basal respiration and SIR-derived micro-
bial carbon. Dehydrogenase activity was measured using
triphenyltetrazolium chloride as a substrate. Samples were
incubated for 16 h at 25  C and the triphenyl formazan pro-
duced was extracted with acetone and measured photometri-
cally at 546 nm [40]. N-mineralisation was determined under
waterlogged conditions according to Kandeler [16]. Water-
logged soils were incubated for seven days at 40  C. The
ammonium released from organic nitrogen compounds was
extracted with 2 M potassium chloride solution, determined
by a modified Berthelot reaction, and measured with a spectro-
photometer at 660 nm [17]. In order to evaluate the nitrifica-
tion capacity of the microbial communities, the potential
nitrification method was used [18]. Soil samples with
233
ammonium sulfate as a substrate and sodium chlorate as an
inhibitor preventing nitrite oxidation were incubated for 5 h
at 25  C and the released nitrite was extracted with 2 M potas-
sium chloride solution and determined colorimetrically at
520 nm. Acid and alkaline phosphomonoesterase activities
were measured using p-nitrophenyl phosphate as a substrate
according to Margesin [24]. Samples were incubated for 1 h at
37  C and the released p-nitrophenol was extracted with 0.5 M
sodium hydroxide in the presence of calcium chloride. The
amounts of extracted products derived from all activity mea-
surements were determined photometrically at 400 nm. All
measured microbiological parameters were calculated on
dry matter bases.
2.4. Statistical analyses
Spearman rank correlation coefficient was used to relate soil
microbiological variables to soil chemical parameters, and to
the amount of mineral fertilizers applied annually and cumu-
latively during the last three years. The Kruskal–Wallis one-
way analysis of variance by ranks’ test was used to assess
the impact of soil type on soil microbiological and chemical
parameters. The data set of soil microbiological variables
was analyzed using principal component analysis (PCA) based
on a correlation matrix, and the effect of binary coded charac-
teristics on the grouping of samples was assessed using
a multivariate randomization test [23] with the computer
program ADE-4 [37]. Prior PCA values of microbiological
variables were log-transformed. The following management
options were binary (presence/absence) coded: use of organic
fertilizers, and pesticides (herbicides, fungicides, and insecti-
cides). The Mann–Whitney test was used to verify the impact
of coded factors on individual soil microbiological variables.
The results of PCA are interpreted using scatters of the sample
scores and a correlation plot showing the relationship of vari-
ables with PCA axes. Grouping of samples due to binary coded
variables is visualised using scatters of the sample scores
connected with group centroids (star plot). Group centroid
coordinates are calculated as the average of the coordinates
of all the group members.
3. Results
Comparison of soil microbiological parameters based on coef-
ficient of variation (CV) showed that the highest variation
among all samples appeared in values of dehydrogenase
activity and potential nitrification (CV 148.2% and 143.0%, re-
spectively), followed by alkaline phosphatase (CV 92.4%), mi-
crobial biomass (CV 89.3%), basal respiration rate (CV 68.8%).
The least variable was acid phosphatase activity (CV 57.9%).
The multivariate randomization test showed that soil type
has a strong effect on soil microbiological variables (P < 0.01,
10,000 permutations). According to the Kruskal–Wallis test,
microbial biomass, activities of dehydrogenase, and alkaline
phosphatase had significantly higher values in Calcaric
Regosols (Table 2). The measured chemical parameters did
not differ between soil types, except for pH (Kruskal–Wallis
test, P < 0.05) the values of which were lowest in Stagnic Luvi-
sols (6.03  0.75), and did not differ between Calcaric Regosols
234 european journal of soil biology 44 (2008) 231–237
Table 2 – Mean values and standard deviations of soil microbiological parameters in different soil types
Soil type (WRB) SIR (mgC g1 dw)
Calcaric Regosols 0.779  0.219**
Calcaric Cambisols 0.559  0.121
Stagnic Luvisols 0.439  0.139
Only parameters that were different among soil types according to Kruskal–Wallis one-way analysis of variance by Ranks are shown. Asterisks
designate group means that are statistically different according to multiple comparisons of mean ranks. *P < 0.05, **P < 0.01.
and Calcaric Cambisols (6.82  0.27 and 6.38  0.35, respec-
tively). Total nitrogen content showed highest variation (CV
106.69%) among samples and was related to soil microbial bio-
mass, dehydrogenase activity, potential nitrification, N-min-
eralisation, and alkaline phosphatase activity (Table 3). Soil
organic matter content correlated significantly with all mea-
sured microbiological parameters, but the relationship was
strongest in the case of potential nitrification, dehydrogenase
activity, N-mineralisation, alkaline and acid phosphatase
activities. The soil pH was related to microbial biomass, basal
respiration, and alkaline phosphatase, but was only slightly
correlated with potential nitrification.
A partial form of principal component analysis was used to
estimate the importance of microbiological variables in the
grouping of the studied fields. In the case of partial PCA, the
analysis was performed on the residual matrix of microbiolog-
ical variables after regression on soil type, i.e. the effect of soil
type was removed from the analysis. The first two principal
components accounted for 78.9% of the total variation in the
microbiological data set. The first principal component was
positively related to dehydrogenase, potential nitrification
and N-mineralisation activity, and microbial biomass, while
the second principal component was negatively correlated
with metabolic quotient and soil respiration (Fig. 1a). Accord-
ing to PCA ordination, the fields of organic farms are located to
the right of the origin and are characterized by higher micro-
bial activity values and biomass (Fig. 1b, Table 4). The second
group of fields is scattered around the origin and consists
mainly of fields that have received both organic and inorganic
fertilizers. The third group of fields, which is situated to the
left of the origin, comprises mostly of fields that in most cases
have received higher amounts of mineral nitrogen as well as
other agrochemicals (herbicides, pesticides and fungicides).
The multivariate randomization test indicated that
Table 3 – Relationships between soil microbiological and
chemical parameters based on Spearman rank
correlation coefficient
Variable Ntot TC
SIR 0.51** 0.45*
Dehydrogenase activity 0.82*** 0.75***
Potential nitrification 0.83*** 0.76***
N-mineralisation 0.62*** 0.60***
Alkaline phosphatase 0.88*** 0.82***
Acid phosphatase 0.50**
Respiration 0.45* 0.39*
*P < 0.05, **P < 0.01, ***P < 0.001.
Dehydrogenase activity Alkaline phospatase
(mgTFP g1 dw 16 h1) (mgNP g1 dw h1)
6.397  4.022* 406.7  185.1*
4.104  0.975 297.5  123.3
2.988  0.705 167.4  157.4
application of organic fertilizers had a significant impact on
the grouping of fields (P < 0.01, 10,000 permutations) (Fig. 1c).
According to the Mann–Whitney test, fields treated with
organic fertilizer are characterized by two times higher micro-
bial biomass (P < 0.01) and N-mineralisation values (P < 0.001).
Data analysis indicated a tendency for increased respiration
and microbial biomass in the case of fields with legume-based
crop rotation (Mann–Whitney test, P < 0.01 and P < 0.05,
respectively). At the same time, these two parameters were
negatively affected by the use of pesticides; respiration
activity (mean values 0.21 and 0.10 mg CO2 24 h1 g1 dw, re-
spectively) was repressed in particular.
Values of studied microbiological variables were regressed
on the amount of mineral fertilizers applied annually and
cumulatively during the last three years. From the set of mea-
sured microbiological parameters, only microbial biomass
was correlated negatively with the amount of mineral nitro-
gen applied annually (R ¼ 0.53, . 0.57, P < 0.01), but the
strongest relationship (R ¼ 0.68, P < 0.001) was found with
the cumulative amount of mineral nitrogen applied during
the last three years.
4. Discussion
Soil type is one of the primary determinants of soil microbial
structure, as demonstrated by polyphasic studies of soil bacte-
rial community composition [13,32]. We found that microbial
biomass, and activities of dehydrogenase and alkaline phos-
phatase are dependent on soil type, whereas measured soil
chemical parameters showed practically no variation among
the three studied soil types. These differences in soil microbial
parameters due to soil type may be related to the qualitative
structure of soil organic carbon as well as to soil texture [33].
In the study of conventional and organic farms Van Diepenin-
gen with co-workers [38] found that soil type in general has
a much stronger effect on the soil characteristics than man-
agement type. Our results support such a conclusion and
stress the importance of considering soil type in data analysis
of soil microbiological variables. In our study we took into
pH
account the effect of soil type in the further statistical analysis
0.69*** of microbiological variables. The partial form of PCA based on
soil biochemical and microbiological parameters separated
0.48**
soils with different management practices, especially soils
0.83***
from organically managed fields. From measured soil bio-
chemical and microbiological parameters, dehydrogenase,
0.80*** potential nitrification, N-mineralisation activity, and micro-
bial biomass contributed most strongly to the separation of
soils in PCA. These microbiological variables are considered
 european journal of soil biology 44 (2008) 231–237 235

Fig. 1 – Partial principal component analysis based on the correlation matrix of soil microbiological parameters. (a)
Ordination of soil samples along first two PCA axes (F1 3 F2). (b) Correlation of soil microbiological parameters with PCA
axes. Abbreviations: AcP – acidic phosphatase, AlP – alkaline phosphatase, D – dehydrogenase, N – N-mineralisation,
PN – potential nitrification, Q – metabolic quantient, R – respiration, SIR – microbial biomass. (c) Star plot showing
impact of manure application on grouping of soil samples. The results of PCA are interpreted using a scatter of the
sample scores connected with group centroids. Group centroids are designated by letters a (no manure used) and
b (fields with manure use).

as potential indicators of soil quality and management impact
in many papers [3,11,12,27,35]. In our case the soils originating
from organically managed farms were generally characterized
by elevated microbiological parameter values, but at the same
time the within-group variation of soil microbiological param-
eters was also highest. The reasons for such large deviations
among organically managed soils may be the different dura-
tions of organic management practice as well as differences
in management history among fields, such as different
amounts and types of organic fertilizers (green or brown
manure) applied and differences in crop rotation.
The application of manure affected the functioning of
the microbial community and two groups were clearly
distinguished: soils which received manure in one or two years
or all the three last years, and those that have not received ma-
nure during the three year period. This treatment had a positive
effect on microbial biomass and N-mineralisation, potential
nitrification, dehydrogenase activity and acidic phosphatase
activity. Surprisingly, there was no significant positive impact
of manure on soil microbial biomass, which has been shown
by several authors [19,21,25]. The consecutive application of
manure over three years enhanced respiration rate, probably
due to the addition of easily degradable organic fractions in
the last year, when soil samples were taken. According to
Sparling [34] and Dilly [9] the quality of organic matter greatly
determines the amount of CO2 efflux from the soil. The

Table 4 – Minimum and maximum values of soil microbiological parameters in soils of studied fields with different
fertilizer use
Variable Only organic Mineral and organic Only mineral
fertilizers (n ¼ 5) fertilizers (n ¼ 6) fertilizers (n ¼ 11)

SIR (mgC g1 dw) 0.46–1.23 0.54–0.85 0.26–0.80

Dehydrogenase activity (mgTFP g1 dw 16 h1) 3.32–14.6 2.96–4.49 2.00–5.41
Potential nitrification (mgN g1 dw 24 h1) 0.85–6.64 0.51–2.91 0.56–4.2
N-mineralisation (mgN g1 dw d1) 0.94–2.28 0.81–1.80 0.40–2.23
Alkaline phosphatase (mgNP g1 dw h1) 92.5–774 90.0–419 20.7–565
Acid phosphatase (mgNP g1 dw h1) 120–336 90–419 118–259
Respiration (mgCO2 24 h1 g1 dw) 0.07–0.24 0.07–0.15 0.05–0.09
236 european journal of soil biology 44 (2008) 231–237
increase of qCO2 due to organic amendments was reported
also by Leita and co-workers [21]. Higher microbial biomass
and activities in the case of organic farming and manure
amendment have been described by many authors
[1,5,6,15,29]. Bittman et al. [4] have found significantly greater
bacterial abundance in the soils where manure has been
used compared to minerally fertilized and untreated soils,
but fungal biomass responded negatively to both types of fertil-
ization. Long-term organic and mineral fertilizer application is
reflected also in altered soil bacterial community structure
[36]. Another management option frequently used for the in-
put of organic matter, legume-based crop rotation, has been
reported to having a positive effect on the microbial commu-
nity [22]. Legume-based crop rotation was also proved to
increase soil respiration and microbial biomass, although the
use of manure had a much stronger overall effect on soil micro-
bial activity.
Strong cumulative negative effect of mineral nitrogen ap-
plication on microbial biomass was revealed in data analysis.
This means that the amount of nitrogen fertilizers added over
time has a stronger effect on microbial biomass than the
amount added in a given year. Such a relationship between
microbial biomass and mineral nitrogen application could be
partly explained by reduced input of readily available organic
matter for soil microbes. Chantigny et al. [7] found that water-
soluble organic carbon contents decreased with increasing
mineral nitrogen fertilizer application due to elevated C-min-
eralisation. Same authors also suggest that increased mineral
nitrogen application may indirectly affect soil respiration by
promoting plant growth and water uptake, which leads to re-
duced soil water content. In our case, simultaneous applica-
tion of manure and mineral fertilizers had the same
negative effect on soil microbial biomass as the use of mineral
fertilizers alone. The amount of mineral nitrogen added over
time was also negatively related to soil acidity, which was
on average 0.4 units lower in soils of minerally fertilized fields.
There was no relationship between the amount of mineral ni-
trogen added and soil organic matter content among the stud-
ied fields. This may be due to the fact that we do not have any
baseline data for the studied fields and, in such a situation,
comparison between fields may not reveal a decrease of soil
organic matter as a result of synthetic N fertilization [20].
Our data did not reveal a negative relationship between the
amount of mineral fertilizer used and dehydrogenase activity,
although Masciandaro et al. [26] have found that mineral and
organic fertilizers affect the kinetic parameters of dehydroge-
nase, substrate affinity and the maximal rate of the enzyme
activity. It could be suggested that the effect on the kinetic
parameters of dehydrogenase might be mediated through
microbial biomass, which decreases with the amendment of
mineral fertilizers; meanwhile the change in substrate affinity
reflects an alteration in soil microbial community structure.
Our results show that changes in Estonian agricultural
management practice during the last decade have already
been reflected in soil microbiological properties. In most
cases, organic farming practice has led to increased microbial
biomass and activities in the soil, but high variation of
microbiological parameters in those soils may, at the same
time, indicate the ongoing transitions caused by changes in
agricultural management practice.
Acknowledgements
This study was part of the research project ‘‘Impact of agricul-
tural management practices to the diversity of soil biota in
Estonian arable soils’’ funded by Estonian Science Foundation
grant No. 5571. The authors are grateful to the head of the
Laboratory of Soil Biology at Hohenheim University, Prof. Ellen
Kandeler, for the opportunity provided for M. Truu to study
soil biochemical methods in their laboratory. We also thank
Dr. Dagmar Tscherco and Dr. Kerstin Mölter for their help
and advice.
references
[1] R. Albiach, R. Canet, F. Pomares, F. Ingelmo, Microbial
biomass content and enzymatic activities after the
application of organic amendments to a horticultural soil,
Bioresource Technol. 75 (2000) 43–48.
[2] T. Beck, R. Öhlinger, A. Baumgarten, Substrate-induced
respiration, in: F. Schinner, R. Öhlinger, E. Kandeler,
R. Margesin (Eds.), Methods in Soil Biology, Springer-Verlag,
Berlin, 1996, pp. 64–68.
[3] G.D. Bending, M.K. Turner, F. Rayns, M.-C. Max, M. Wood,
Microbial and biochemical soil indicators and their
potential for differentiating areas under contrasting
agricultural management regimes, Soil Biol. Biochem. 36
(2004) 1785–1792.
[4] S. Bittman, T.A. Forge, C.G. Kowalenko, Response of the
bacterial and fungal biomass in a grassland soil to multi-year
applications of dairy manure slurry and fertilizer, Soil Biol.
Biochem. 37 (2005) 613–623.
[5] L.R. Bulluck, M. Brosius, G.K. Evanylo, J.B. Ristaino, Organic
and synthetic fertility amendments influence soil microbial,
physical and chemical properties on organic and
conventional farms, Appl. Soil Ecol. 19 (2002) 147–160.
[6] L. Böhme, U. Langer, F. Böhme, Microbial biomass, enzyme
activities and microbial community structure in two
European long-term field experiments, Agric. Ecosyst.
Environ. 109 (2005) 141–152.
[7] M.H. Chantigny, D.A. Angers, D. Prevost, R.R. Simard, F.-P.
Chalifour, Dynamics of soluble organic C and C
mineralisation in cultivated soils with varying N fertilization,
Soil Biol. Biochem. 31 (1999) 543–550.
[8] R.P. Dick, Soil activities as integrative indicators of soil
health, in: C. Pankhurst, B.M. Doube, V.V.S.R. Gupta (Eds.),
Biological Indicators of Soil Health, CABI Publishing, 1997, pp.
121–156.
[9] O. Dilly, Microbial respiratory quotient during basal
metabolism and after glucose amendment in soils and litter,
Soil Biol. Biochem. 33 (2001) 117–127.
[10] O. Dilly, H.-P. Blume, J.C. Munch, Soil microbial activities in
Luvisols and Anthrosols during 9 years of region-typical
tillage and fertilisation practices in northern Germany,
Biogeochemistry 65 (2003) 319–339.
[11] Z. Filip, International approach to assessing soil quality by
ecologically-related biological parameters, Agric. Ecosyst.
Environ. 88 (2002) 169–174.
[12] F. Gil-Sotres, C. Trasar-Cepeda, M.C. Leirós, S. Seoane,
Different approaches to evaluating soil quality using
biochemical properties, Agric. Ecosyst. Environ. 37 (2005)
877–887.
[13] M.S. Girvan, J. Bullimore, J.N. Pretty, A.M. Osborn, A.S. Ball,
Soil type is the primary determinant of the composition of
 european journal of soil biology 44 (2008) 231–237
the total and active bacterial communities in arable soils,
Appl. Environ. Microbiol. 69 (2003) 1800–1809.
[14] R.L. Haney, S.A. Senseman, L.J. Krutz, F.M. Hons, Soil carbon
and nitrogen mineralisation as affected by atrazine and
glyphosate, Biol. Fertil. Soils 35 (2002) 35–40.
[15] D.G. Hole, A.J. Perkins, J.D. Wilson, I.H. Alexander, P.V. Grice,
A.D. Evans, Does organic farming benefits biodiversity? Biol.
Conserv. 122 (2005) 113–130.
[16] E. Kandeler, N-mineralisation under waterlogged conditions,
in: F. Schinner, R. Öhlinger, E. Kandeler, R. Margesin (Eds.),
Methods in Soil Biology, Springer-Verlag, Berlin, 1996, pp.
141–143.
[17] E. Kandeler, Ammonium, in: F. Schinner, R. Öhlinger, E.
Kandeler, R. Margesin (Eds.), Methods in Soil Biology,
Springer-Verlag, Berlin, 1996, pp. 406–408.
[18] E. Kandeler, Potential nitrification, in: F. Schinner, R.
Öhlinger, E. Kandeler, R. Margesin (Eds.), Methods in Soil
Biology, Springer-Verlag, Berlin, 1996, pp. 147–149.
[19] E. Kandeler, M. Stemmer, M.-M. Klimanek, Response of soil
microbial biomass, urease and xylanase within particle size
fractions to long-term soil management, Soil Biol. Biochem.
31 (1999) 261–273.
[20] S.A. Khan, R.L. Mulvaney, T.R. Ellsworth, C.W. Boast, The
myth of nitrogen fertilization for soil carbon sequestration, J.
Environ. Qual. 36 (2007) 1821–1832.
[21] L. Leita, M. De Nobili, C. Mondini, G. Muhlbachova,
L. Marchiol, G. Bragato, M. Cotin, Influence of inorganic and
organic fertilization on soil microbial biomass, metabolic
quotient and heavy metal bioavailability, Biol. Fertil. Soils 28
(1999) 371–376.
[22] N.Z. Lupwayi, W.A. Rice, G.W. Clayton, Soil microbial
diversity and community structure under wheat as
influenced by tillage and crop rotation, Soil Biol. Biochem. 30
(1998) 1733–1741.
[23] B.F.J. Manly, Randomization, Bootstrap and Monte Carlo
Methods in Biology, Chapman and Hall, London, 1997, pp.
172–204.
[24] R. Margesin, Acidic and alkaline phosphomonoesterase
activity with the substrate p-nitrophenyl phosphate, in:
F. Schinner, R. Öhlinger, E. Kandeler, R. Margesin (Eds.),
Methods in Soil Biology, Springer-Verlag, Berlin, 1996, pp.
213–217.
[25] P. Marschner, E. Kandeler, B. Marschner, Structure and
function of soil microbial community in a long-term
fertilizer experiment, Soil Biol. Biochem. 35 (2003) 453–461.
[26] G. Masciandaro, B. Ceccanti, V. Ronchi, C. Bauer, Kinetic
parameters of dehydrogenase in the assessment of the
response of soil to vermicompost and inorganic fertilisers,
Biol. Fertil. Soils 32 (2000) 479–483.
[27] P. Mäder, A. Fließbach, D. Dubois, L. Gunst, P. Fried, U. Niggli,
Soil fertility and biodiversity in organic farming, Science 296
(2002) 1694–1697.
237
[28] R.L. Parfitt, G.W. Yeates, D.J. Ross, A.D. Mackay, P.J.
Budding, Relationships between soil biota, nitrogen and
phosphorus availability, and pasture growth under organic
and conventional management, Appl. Soil Ecol. 28 (2005)
1–13.
[29] J.A. Parham, S.P. Deng, W.R. Raun, G.V. Johnson, Long-term
cattle manure application in soil. I. Effect on soil
phosphorus level, microbial biomass C, and dehydrogenase
and phosphatase activities, Biol. Fertil. Soils 35 (2002)
328–337.
[30] D.S. Powlson, The soil microbial biomass: before, beyond and
back, in: K. Ritz, J. Dighton, G.E. Giller (Eds.), Beyond the
Biomass, Wiley, Chichester, UK, 1994, pp. 3–20.
[31] M. Schloter, O. Dilly, J.C. Munch, Indicators for evaluating soil
quality, Agric. Ecosyst. Environ. 98 (2003) 255–262.
[32] D. Seghers, S.D. Siciliano, E.M. Tot, W. Verstraete, Combined
effect of fertilizer and herbicide applications and the
abundance, community structure and performance of the
soil methanothrophic community, Soil Biol. Biochem. 37
(2005) 187–193.
[33] A.V. Spain, R.F. Isbell, M.E. Propert, Soil organic matter, in:
Soils: Australian Viewpoint, CSIRO/Academic Press,
Melbourne/London, 1983, pp. 551–563.
[34] G.P. Sparling, Soil microbial biomass, activity and nutrient
cycling as indicators of soil health, in: C. Pankhurst, B.M.
Doube, V.V.S.R. Gupta (Eds.), Biological Indicators of Soil
Health, CABI Publishing, 1997, pp. 97–120.
[35] G.P. Sparling, L.A. Schipper, W. Bettjeman, R. Hill, Soil quality
monitoring in New Zealand: practical lessons from a 6-year
trial, Agric. Ecosyst. Environ. 104 (2004) 523–534.
[36] H.Y. Sun, S.P. Deng, W.R. Raun, Bacterial community
structure and diversity in a century-old manure-treated
agroecosystem, Appl. Environ. Microbiol. 70 (2004) 5868–
5874.
[37] J. Thioulouse, D. Chessel, S. Doledec, J.-M. Olivier, ADE-4:
a multivariate analysis and graphical display software, Stat.
Comput. 7 (1997) 75–83.
[38] A.D. Van Diepeningen, O.J. de Vos, G.W. Korthals, A.H.C. van
Bruggen, Effects of organic versus conventional
management on chemical and biological parameters in
agricultural soils, Appl. Soil Ecol. 31 (2006) 120–135.
[39] K. Watanabe, N. Hamamura, Molecular and physiological
approaches to understanding the ecology of pollutant
degradation, Curr. Opin. Biotechnol. 14 (3) (2003)
289–295.
[40] R. Öhlinger, Dehydrogenase activity with the substrate TTC,
in: F. Schinner, R. Öhlinger, E. Kandeler, R. Margesin (Eds.),
Methods in Soil Biology, Springer-Verlag, Berlin, 1996, pp.
241–243.
[41] R. Öhlinger, Soil respiration by titration, in: F. Schinner,
R. Öhlinger, E. Kandeler, R. Margesin (Eds.), Methods in Soil
Biology, Springer-Verlag, Berlin, 1996, pp. 95–98.