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Waste Management 28 (2008) 1246–1253

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Changes in soil chemical and microbiological properties during

4 years of application of various organic residues
a,*
M. Odlare , M. Pell b, K. Svensson b

a
Department of Public Technology, Mälardalen University, Box 883, SE-721 23 Västerås, Sweden
b
Department of Microbiology, Swedish University of Agricultural Sciences, Box 7025, SE-750 07 Uppsala, Sweden

Accepted 19 June 2007

Available online 13 August 2007

Abstract

A 4-year field trial was established in eastern Sweden to evaluate the effects of organic waste on soil chemical and microbiological vari-
ables. A simple crop rotation with barley and oats was treated with either compost from household waste, biogas residue from household
waste, anaerobically treated sewage sludge, pig manure, cow manure or mineral fertilizer. All fertilizers were amended in rates corresponding
to 100 kg N ha1 year1. The effects of the different types of organic waste were evaluated by subjecting soil samples, taken each autumn 4
weeks after harvest, to an extensive set of soil chemical (pH, Org-C, Tot-N, Tot-P, Tot-S, P-AL, P-Olsen, K-AL, and some metals) and
microbiological (B-resp, SIR, lSIR active and dormant microorganisms, PDA, lPDA, PAO, Alk-P and N-min) analyses. Results show that
compost increased pH, and that compost as well as sewage sludge increased plant available phosphorus; however, the chemical analysis
showed few clear trends over the 4 years and few clear relations to plant yield or soil quality. Biogas residues increased substrate induced
respiration (SIR) and, compared to the untreated control amendment of biogas residues as well as compost, led to a higher proportion
of active microorganisms. In addition, biogas residues increased potential ammonia oxidation rate (PAO), nitrogen mineralization capacity
(N-min) as well as the specific growth rate constant of denitrifiers (lPDA). Despite rather large concentrations of heavy metals in some of the
waste products, no negative effects could be seen on either chemical or microbiological soil properties. Changes in soil microbial properties
appeared to occur more rapidly than most chemical properties. This suggests that soil microbial processes can function as more sensitive
indicators of short-term changes in soil properties due to amendment of organic wastes.
Ó 2007 Elsevier Ltd. All rights reserved.

1. Introduction Compost from organic waste is frequently mixed with

Organic waste can be a valuable and inexpensive soil
conditioner and source of plant nutrients, and positive
effects of organic waste on soil have been reported in sev-
eral studies. For example, organic waste application can
increase soil nitrogen and phosphorous contents (Jakob-
sen, 1995), improve soil structure and water holding capac-
ity (Joshua et al., 1998), increase microbial biomass
(Leifeld et al., 2002; Jedidi et al., 2004), reduce the need
for chemical weed control (Pinamonti, 1998) and suppress
plant diseases (Hoitink and Boehm, 1999).
*
Corresponding author. Tel.: +46 21 101611; fax: +46 21 101370.
E-mail address: monica.odlare@mdh.se (M. Odlare).
urban garden waste in order to improve its structure and
to increase the C:N ratio. Due to anthropogenic activities,
such as leaded fuel in the past, park and garden waste often
contain heavy metals. Sewage sludge is known to some-
times contain heavy metals and the question of whether
agricultural soils should be amended with sewage sludge
or not has been debated since the late 1960s. Biogas can
be produced from a wide variety of organic wastes, which
may result in both different and variable qualities of the
resulting residue. Anaerobic digestate from source sepa-
rated household waste may contain several types of organic
contaminants (Nilsson et al., 2000). Hence, there is an
urgent need for long-term field experiments to evaluate
the effects of repeated amendment of organic waste on soil
properties.

0956-053X/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.wasman.2007.06.005
 M. Odlare et al. / Waste Management 28 (2008) 1246–1253
Soil quality is a complex characteristic determined by
physical, chemical and biological components of the soil.
Doran and Parkin (1994) define soil quality as ‘‘the capac-
ity of a soil to sustain biological productivity, maintain
environmental quality, and promote plant and animal
health’’. Changes in the soil microbial community may
occur more rapidly than changes in other soil characteris-
tics and therefore, soil microbial processes are thought to
be sensitive indicators of changes in soil quality (Kennedy
and Papendick, 1995). In a well-designed field experiment,
the accumulated effect of organic waste on soil quality can
be monitored and evaluated during a long period under
realistic conditions.
The objective of this study was to evaluate the effects of
application of organic waste on soil chemical and microbi-
ological properties in an agricultural soil. A 4-year field
experiment was conducted to compare the effects of com-
post from municipal household waste, biogas residues
and anaerobically treated sewage sludge with the tradi-
tional fertilizers cow manure, liquid pig manure and min-
eral fertilizer (NPS). Soil samples were collected each
year and the changes in different chemical and microbio-
logical soil properties were monitored in order to evaluate
the effect on soil quality. Another objective was to compare
the sensitivity of soil chemical and microbiological param-
eters to function as indicators of soil quality in terms of
reflecting changes in soil properties.
2. Materials and methods
2.1. Site characteristics and experimental design
The field experiment was established in autumn 1998 at
a site located in central Sweden (59°37 0 N, 16°33 0 E). The
soil had not been fertilized with farmyard manure since
1975, and in some 20 years before the start of the experi-
ment the field had been cropped mainly with cereals, oil
seed rape and legumes. Soil management was conventional
with annual plowing. The soil contained 37–49% clay, 36–
44% silt and 14–20% sand, and was classified as a Eutric
Cambisol (FAO, 1998). The experiment was set up in a
random block design with four replications and a plot size
of 90 m2. The crop rotation in the experiment comprised
oats (Avena sativa cv. Freja) and spring barley (Hordeum
vulgare cv. Baronessa). The site for the field experiment
was selected using a specific selection technique where near
infrared reflection (NIR) spectroscopy is combined with
principal component analysis (Odlare et al., 2005). The
NIR strategy made it possible to find the area within the
field that had the smallest spatial variation of soil
properties.
2.2. The fertilizers
All organic and inorganic fertilizers were applied at a
rate corresponding to 100 kg N ha1 year1. The compost
(C) and the liquid biogas residues (BR) were applied at
1247
two rates: C100 and C50, and BR100 and BR50, respec-
tively. In C100 and BR100, all N originated from the
organic waste, whereas in C50 and BR50 half of the N
was supplied as mineral N. Anaerobically treated sewage
sludge (SS), pig manure (PM) and cow manure (CM) were
all complemented with 50 kg mineral N ha1 year1. Other
plots had solely mineral fertilizer (NPS) applied at
100 kg N ha1 year1. The reason for using NPS instead
of NPK, was that the soil in the field experiment is rich
in potassium but poor in sulphur. In addition, there is a
general trend towards lower atmospheric sulphur deposi-
tion, which leads to a need for complementary sulphur fer-
tilization. The nitrogen in the mineral fertilizer was 50% in
the NH4 form and 50% in the NO3 form. Unfertilized plots
were used as control. The treatments are summarized in
Table 1, and the actual applied yearly amounts of dry mat-
ter and various plant nutrients are given in Table 2. In
order to be able to apply the results to real agricultural sys-
tems, the different types of manure were applied according
to general farming practice in Sweden. Therefore, compost,
sewage sludge and cow manure were spread a few days
before ploughing in late autumn. Biogas residues and pig
manure were spread on the seedlings immediately before
stem elongation. Mineral fertilizers were applied in spring
at sowing. Compost, sludge, cow manure and mineral fer-
tilizer were applied by hand from a bucket as evenly as pos-
sible. Biogas residue and pig manure were spread by hand
with a watering can.
The compost was produced at the municipal composting
plant at Västerås, close to the study site. Source separated
household waste (70%) was mixed with chopped park and
garden litter (30%) before composting. The biogas residue
was obtained from a biogas plant in Stockholm where
source-separated household waste is co-digested with food
residues from restaurants and kitchens. The sewage sludge
was taken from the municipal wastewater treatment plant
in the city of Västerås. The plant treats sewage from
118,000 p.e. yielding 12,000 tonnes dewatered (25% d.m.)
sludge per year. Influent raw wastewater is pre-precipitated
with iron sulphate, biologically treated by an activated
sludge process with pre-denitrification supported with gly-
col as external carbon and energy source. Excess sludge is
anaerobically digested to reduce its volume, make the
product hygienic and produce biogas. The pig and cow
manure was collected from local farmers.
2.3. Soil sampling
Soil samples were collected separately in each plot prior
to the experiment and then every year in autumn 4 weeks
after harvest. Each experimental plot was sampled about
25 times from the topsoil at a 0–20 cm depth with soil cor-
ers (diameter 22–25 mm) to obtain 4 kg of moist soil per
plot. The soil samples were put in polythene bags and
transported to the laboratory the same day where
they were stored at +2 °C. In order to be manageable,
1248 M. Odlare et al. / Waste Management 28 (2008) 1246–1253
Table 1
The different treatments used in the ORC field experiment
Treatment Residue/fertilizer N in organic
fertilizer
(kg ha1 year1)
C100 Compost 100
C50 Compost + mineral N 50
BR100 Biogas residues 100
BR50 Biogas residues + mineral N 50
SS Sewage sludge + mineral N 50
PM Pig manure + mineral N 50
CM Cow manure + mineral N 50
NPS NPS 0 100
Control No fertilizer 0 0
All treatments correspond to a total of 100 kg N ha1 year1.
a
The time for application differed between the 4 years. These dates represent the earliest and the latest date.
the samples were dried slowly at +2 °C to a dry matter con-
tent of 83%. For biological analyses each sample was sieved
(5 mm), thoroughly mixed and portioned in polythene bags
for the different analyses. The samples were stored at
20 °C and all analyses were performed within 13 months
of the sampling date (Stenberg et al., 1998). All biological
tests were run in triplicate. For ammonium and nitrate
analyses, the samples were frozen moist at arrival to the
laboratory whereas for the remaining chemical analyses,
the samples were dried (35 °C) and sieved (2 mm).
2.4. Chemical and physical analyses
Total soil carbon (Tot-C), nitrogen (Tot-N) and sulphur
(Tot-S) were determined after combustion at 1250 °C with
a carbon, nitrogen and sulphur analyzer (CNS-2000,
LECO Equipment Corp., St. Joseph, MI, USA). Tot-C
was corrected for carbonate to give organic C (Org-C).
Mineral N (NH4-N and NO3-N) was analyzed with dialysis
on an AutoAnalyzer TRAACS 800 (Kontram, Stockholm,
Sweden). NH4-N was analyzed according to the method
ST9002-NH4D and NO3-N according to the method
ST9002-NO3D. Organic N was measured by the Kjeldahl
method (Bremner and Mulvaney, 1982). Available P
(P-AL) and K (K-AL) were extracted with the ammonium
lactate method (AL) and analyzed as described by Egnér
et al. (1960). Available P (P-Ols) was also analyzed accord-
ing to the method described by Olsen et al. (1954). Total
cation-exchange capacity (CEC) and total exchangeable
base cations (TEB) were determined according to Nömmik
(1974). The pH was determined in 0.01 M CaCl2 with the
vol/vol ratio 1:2 (pHCaCl2). Total sulphur (Tot-S) and the
metals Cu, Zn, Cd, Ni, Pb and Cr were analyzed by ICP-
AES (Perkin-Elmer ICP Optima 3000) according to SS
02 83 11 (SIS, 1997). The particle size distribution was
determined according to Jung (1987).
2.5. Microbiological analysis
Microbial biomass measured as substrate induced respi-
ration (SIR), specific growth rate (lSIR) and the basal
Applied N in mineral Time for applicationa
fertilizer (kg ha1 year1)
0 6 November to 3 December
50 6 November to 3 December
0 16 June to 29 June
50 16 June to 29 June
50 6 November to 3 December
50 16 June to 29 June
50 6 November to 3 December
25 April to 20 May
respiration (B-resp), were determined in 25 g soil adjusted
to 50% of the water holding capacity and incubated at
20°C. CO2 evolution was determined hourly with a respi-
rometer (Respicond III, Nordgren Innovations AB, Umeå,
Sweden). The CO2 was determined from conductivity
changes measured during accumulation in a 250 mM
KOH solution (Nordgren, 1992). B-resp was calculated
as the average respiration rate measured for incubation
times between 200 and 240 h. SIR measurement was then
initiated by mixing a substrate consisting of 75 mg glucose,
2.5 mg (NH4)2SO4-N, 0.5 mg KH2PO4-P and 1000 mg of
powdered talcum into each soil sample. Empty vessels were
used as control. The respiration data were fitted by non-
linear regression to an exponential growth equation as
described by Stenström et al. (2001):
dp=dt ¼ relt þ K ð1Þ
where dp/dt is the rate of CO2 formation, r is the initial res-
piration rate of growing microorganisms, l is the specific
growth rate, and t is the time after addition of the SIR sub-
strate. Hence, relt is the CO2 production rate of exponen-
tially growing microorganisms and K is the production
rate of non-growing microorganisms. The SIR rate is de-
fined as the sum of r and K, i.e., the total respiration rate
obtained instantaneously on the addition of the SIR sub-
strate. Thus, by use of the equation, the SIR response after
addition of glucose can be divided into the initial respira-
tion rate of growing (active) and non-growing (dormant)
microorganisms. Altogether, the method yields a quantita-
tive estimation of the active and dormant microorganisms,
as well as the parameters SIR and lSIR.
The potential ammonia oxidation rate (PAO) in 25 g of
soil was assayed as accumulated nitrite according to the
short-incubation chlorate-inhibition technique described
by Belser and Mays (1980) and modified by Torstensson
(1993); ISO 15685 (2004). The rate of NO 2 formation
was determined by linear regression.
The potential denitrification activity (PDA) was assayed
in 25 g of soil according to the modified short incubation
C2H2-inhibition method described by Pell et al. (1996).
The assay substrate contained 2 mM glucose and 2 mM
KNO3. During the assay, seven gas samples were
 M. Odlare et al. / Waste Management 28 (2008) 1246–1253 1249

Table 2
Application rates of dry matter, total carbon, plant nutrients and heavy metals in the field experiment
kg ha1 year1 C50 C100 BR50 BR100 SS PM CM NPS
Dry matter 2520 5040 287 575 1335 559 2252 366
Tot-C 565 1130 103 207 324 203 743 0
Org-N 47 94 20 39 42 14 43 0
NH4-N 0 0 30 61 8 36 7 50
NO3-N 3 6 0 0 0 0 0 50
Supplemental mineral N 50 0 50 0 50 50 50 100
P-Olsen 0.5 0.9 2.1 4.3 0.4 12 5 14.8
P-AL 5.9 11.8 2.3 4.7 6 13 11 15.4
K-AL 20 39 21 42 1 34 68 0.8
Tot-P 11 22 3.5 6.9 37 22 17 20
Tot-S 6.7 13.4 2.2 4.3 12 8 10 13.7
Cua 170 340 17 35 433 140 76 1
Zna 520 1030 57 110 620 631 415 9
Cda 1 2.1 0.1 0.2 0.9 0.3 0.4 0.04
Nia 22 44 3 6 15 4 7 3
Pba 58 120 3 5 21 1 4 0.2
Cra 40 81 7 13 18 17 6 12
Total carbon (Tot-C), organic nitrogen (Org-N), ammonium (NH4-N) and nitrate (NO3-N), available P (P-Olsen), available P (P-AL), available K
(K-AL), total P (Tot-P), total S (Tot-S). The values represent mean values for 4 years (n = 4). For treatment abbreviations, see Table 1.
a
Expressed as g ha1 year1.

withdrawn from the headspace and analyzed for N2O on a
gas chromatograph provided with an electron capture
detector. The rate of N2O formation increased with time
and the data were fitted to a product formation equation
that takes exponential growth into consideration (Sten-
ström et al., 1991). Two parameters, the initial product for-
mation rate (PDA) and the specific growth rate (lPDA),
were derived by non-linear regression.
Alkaline phosphatase activity (Alk-P) was assayed
according to the p-nitrophenol (PNP) method (Sjöqvist,
1993). Prior to the analysis, the soils were preincubated
for 4 weeks at 15 °C and 50% WHC. After this incubation
period, 2 g of soil was mixed in a flask with 2 ml of water,
8 ml 0.1 M TRIS (pH 9.0) and a substrate of 2 ml 0.115 M
PNP-P. The rate of product formation was determined as
the difference in concentration of PNP between the start
and end of a 2-h incubation period.
The nitrogen mineralization capacity (N-min) was mea-
sured with the anaerobic incubation method described by
Waring and Bremner (1964) and modified by Stenberg
et al. (1998). A slurry with 10 g of soil was anaerobically
incubated for 10 days at 37 °C before extraction and anal-
ysis of NHþ þ
4 . The rate of NH4 formation was determined
as the difference in product between the start and end of
a 10-day incubation period.
2.6. Data treatment
Descriptive statistics are presented as maximum, mini-
mum, mean, median and coefficient of variance (CV). Sig-
nificant effects between the different treatments were
evaluated by one-way ANOVA followed by Tukey’s
HSD multiple comparison test. Statistical interactions
between treatment and year were evaluated using a general
linear model. All statistical tests were performed by
the computer software package SPSS 11.5.1, SPSS Inc.,
Chicago, IL, USA.
3. Results
3.1. Organic fertilizers
Selected characteristics of the fertilizers used in this
study are presented in Table 2. More than half of the nitro-
gen content of the biogas residue occurred as ammonium,
whereas only small amounts of mineral nitrogen, mainly
nitrate, were found in compost. In sewage sludge, pig man-
ure and cow manure, the mineral nitrogen was present as
ammonium. Biogas residues contained small amounts of
both total phosphorous (Tot-P) and plant available phos-
phorous (P-AL), whereas sewage sludge was high in Tot-
P but low in P-AL. High concentrations of plant available
potassium (K-AL) were found in all organic fertilizers
except sewage sludge. The compost contained large con-
centrations of all heavy metals analyzed, especially Zn
and Pb. Sewage sludge contained large amounts of Cu.
Plots amended with compost as the sole fertilizer exceeded
the Swedish Environmental Protection Agency’s threshold
values for all heavy metals, whereas plots amended with
sewage sludge exceeded the threshold value for Cu. Since
the application rate was based on nitrogen supply, the
dry matter applied in the compost-amended plots was
nearly nine times higher than the biogas residue-amended
plots and twice as high as the sewage sludge-amended
plots.
1250 M. Odlare et al. / Waste Management 28 (2008) 1246–1253
3.2. Statistics
The soil nutrient, metal and microbial properties prior
to the start of the experiment are shown in Table 3. The
values represent mean values for the entire experiment
area, i.e., mean values of all plots (n = 36). The CV-values
indicate only a small spatial variation at the site, and hence
treatment-effects should not be obscured by intrinsic
variation.
Analyses with ANOVA showed no significant differ-
ences between the years and no interactions between treat-
ment and year. Also, there was no significant difference
between years 1 and 4. Therefore, the accumulated effects
(Table 4) are presented as mean values for all 4 years
(n = 16). This has the advantage that effects of extreme
meteorological conditions during specific years are
avoided. Data interpretation was supported by Tukey
HSD (p = 0.10). A relatively high probability value was
chosen because a field experiment always involves a high
risk of environmental disturbances where rainfall, drought,
temperature, etc. may obscure treatment effects. Therefore,
in order to be able to more accurately detect changes due to
the different treatments, the significance level was set to
p = 0.10.
3.3. The crop
Crop yields and grain quality in the ORC-experiment
are discussed in detail in Svensson et al. (2004). Crop yield
data is summarized in Table 4.
3.4. Soil chemical properties
The results of all chemical analyses are presented in
Table 4. Significant differences between the treatments
could only be detected for three of the soil chemical analy-
ses made. C100 and PM resulted in a small, though signif-
icant, increase in pH by approximately 0.2 units compared
to NPS and the untreated control. C100 and SS increased
the level of P-AL in the soil by about 20% compared to
BR50, NPS and the control treatment. Although not signif-
icant, a trend of increasing P-AL concentrations was
observed in C50 and BR100. K-AL was about 9% larger
in plots applied with C50, C100, PM, CM and BR100 com-
pared to the NPS and untreated control.
3.5. Soil microbiological properties
The results of all microbiological analyses are presented
in Table 4. None of the fertilizers had any clear effects on
B-resp, but BR50 and BR100 increased substrate-induced
respiration (SIR) by about 11% compared to the untreated
control. Generally, the SIR response consisted of a biomass
with about 90% dormant and 10% active organisms. Com-
pared with the untreated control, the proportion of active
biomass was about 25% higher in C50, BR50, PM and
NPS, but was only slightly larger in C100 and SS. The dor-
mant microbial biomass was significantly larger in BR100
than NPS and the untreated control. None of the fertilizers
had any significant effect on the potential denitrification
rate (PDA). However, the specific growth rate constant
lPDA was about 19% larger in BR100 compared to
NPS. The potential nitrification rate (PAO) showed a sim-
ilar pattern as lPDA, being approximately 19% higher in
B100 than in NPS. CM and BR100 increased nitrogen min-
eralization (N-min) compared to NPS and the untreated
plot by about 20%.
4. Discussion
4.1. Soil chemical properties
Of all of the chemical soil properties analyzed, only
three of them showed significant differences between the
treatments: pH, K-AL and P-AL.
Compost increased the pH in the soil, which could prob-
ably be explained by the production of ammonia formed
as the organically bound nitrogen is mineralized and then
transformed into ammonia in the aerated soil. Similar
results have been reported by Jakobsen (2005) and Lee
et al. (2004). The increased pH in the plots fertilized with
pig manure can be explained by the relatively high concen-
trations of ammonium nitrogen in this fertilizer.
Plant available phosphorous (P-AL) increased in the
plots amended with compost and sewage sludge. Both these
fertilizers were rich in phosphorous but poor in nitrogen,
and since the application rate was based on nitrogen con-
tent, the amount of phosphorous in the soil is likely to
increase.
The same reasoning applies for plant available potas-
sium (K-AL). Application of fertilizers rich in potassium
(compost, biogas residues, pig manure and cow manure)
increased the content in the soil.
4.2. Soil microbiological properties
The majority of the microbial populations in the soil are
heterotrophs and therefore dependent on the availability of
organic carbon and energy, such as sugar and starch.
Hence, amending the soil with organic carbon should
support a larger biomass and thereby affect respiration
parameters. SIR responded well to the application of bio-
gas residues, which was expected, because it is likely to con-
tain a higher amount of easy-degradable carbon than, for
example, compost. Degradation of organic material is less
effective in the anaerobic biogas process and proceeds for a
shorter period of time than aerobic degradation through
composting. In addition, the ammonium content in biogas
residue could have contributed to the increased plant
growth with higher yields (Svensson et al., 2004), which
in turn would supply more carbon to the soil through litter
and root exudates.
The proportion of active microorganisms in the soil was
generally small, only about 10%. This is in agreement with
 M. Odlare et al. / Waste Management 28 (2008) 1246–1253 1251

Table 3
Descriptive statistics (maximum, minimum, mean, median and CV) for various soil properties for the experimental site prior to the experiment (n = 36)
Parameter Range Mean Median CV
Minimum Maximum
Chemical analysis
pH 5.4 5.7 5.6 5.6 1.4
Org-C (%) 1.2 1.6 1.3 1.3 6.3
Tot-C (%) 1.2 1.6 1.3 1.3 6.1
Tot-N (%) 0.1 0.2 0.1 0.1 6.3
Tot-P (g kg1 dw) 0.6 0.7 0.7 0.7 6.3
Tot-S (g kg1 dw) 0.2 0.2 0.2 0.2 4.3
P-AL (mg 100 g1 dw) 3.4 5.4 4.6 4.8 13.0
P-Olsen (mg 100 g1 dw) 1.7 2.9 2.2 2.2 10.6
K-AL (mg 100 g1 dw) 16.5 21.0 18.5 18.5 6.1
TEB (mekv 100 g1) 10.0 12.6 11.2 11.2 6.0
CEC (mekv 100 g1) 15.8 20.2 17.1 16.8 5.3
Cu (mg kg1 dw) 22.1 31.3 25.7 26.0 9.5
Zn (mg kg1 dw) 81.6 98.4 90.3 91.6 5.9
Cd (mg kg1 dw) 0.2 0.3 0.3 0.3 6.0
Ni (mg kg1 dw) 20.0 28.5 24.0 23.8 11.2
Pb (mg kg1 dw) 19.1 23.3 20.4 20.4 3.9
Cr (mg kg1 dw) 34.2 42.2 37.2 36.6 51.2
Soil analysis (% of mineral fraction)
>0.002 mm 37.0 49.0 42.1 41.5 7.4
0.02–0.002 mm 24.0 29.0 26.5 27.0 4.9
0.06–0.02 mm 11.0 18.0 14.6 15.0 12.9
0.2–0.06 mm 13.0 19.0 15.5 15.0 10.6
Microbial analysis
B-resp (lg CO2-C g1 dw h1) 1.2 0.7 0.9 0.9 12.9
SIR (lg CO2-C g1 dw h1) 11.4 15.4 13.4 13.5 6.4
lSIR (min1) 0.12 0.16 0.15 0.15
PDA (ng N2O-N g1 dw min1) 7.1 11.5 9.1 8.9 11.6
lPDA (min1) 0.002 0.005 0.004 0.004 18.2
PAO (ng NO2-N g1 dw min1) 1.6 3.9 2.9 2.8 17.2
Alk-P (lmol PNP g1 dw h1) 1.1 1.8 1.5 1.5 9.3
1
N-min (lgNHþ 4 -N g dw 10 d1) 19.1 24.0 24.4 21.2 7.2
Total carbon (Tot-C), total N (Tot-N), total exchangeable base cations (TEB), cation exchange capacity (CEC), basal respiration (B-resp), substrate-
induced respiration (SIR), growth rate constant (lSIR), potential denitrification activity (PDA), growth rate constant (lPDA), potential ammonia
oxidation (PAO), alkaline phosphatase activity (Alk-P), N-mineralization capacity (N-min). For additional abbreviations, see Table 2.
dw, dry weight.

the 5–10% active organisms as reported by Stenström et al.
(2001) for soils with no recent addition of organic sub-
strate. Moreover, Stenström et al. (2001) observed that
up to 100% of the microbial biomass can be active in soils
with high substrate availability, followed by periods of sev-
eral weeks of back transition into a dormant state when
substrate ceased. One reason for the low proportion of
active organisms found in our field experiment most likely
was that the soil sampling took place 4 weeks after harvest,
a stage where most easy available carbon remaining after
harvest had been consumed. The largest proportion of
active microorganisms was found in BR50 and NPS, which
were also the treatments that gave the largest crop yields.
The results suggest that proportion of active microorgan-
isms can be an indicator of the amount of easy degradable
carbon in the soil. It should also be noted that when long-
term changes in soil quality is to be assessed, sampling
should preferably be performed at occasions when the soil
microorganisms and plant roots are not in an active grow-
ing state. Our strategy is to collect samples 1 month after
harvest seemed to meet such a criterion.
Denitrifiers are heterotrophs and although PDA showed
no significant treatment effects, the enzymes reflected in
growth, lPDA, increased as a result of application of bio-
gas residues. The bacteria involved in denitrification usu-
ally respond quickly to the presence of easy degradable
carbon (deCantanzaro and Beuchamp, 1985). Although
denitrifiers are facultative anaerobes respiring both under
aerobic and anaerobic conditions using carbon as an
energy source, they grow faster in the presence of oxygen.
Intense aerobic respiration will successively lower the par-
tial pressure of oxygen thus stimulating anaerobic denitrifi-
cation activity. Hence, sources of readily degradable
carbon are important for denitrifying bacteria and their
activities. Altogether, the carbon-dependent microbial
parameters SIR, dormant and active microorganisms and
lPDA seem to be efficient indicators of available carbon
in the soil.
1252 M. Odlare et al. / Waste Management 28 (2008) 1246–1253

Table 4
Soil chemical and microbiological characteristics and crop yield after fertilizer amendment
C50 BR50 SS PM CM NPS C100 BR100 Control
bc a c
pH 5.7 5.7 5.7 5.7 5.7 5.6 5.7 5.7 5.6ab
Org-C (%) 1.3 1.3 1.3 1.3 1.3 1.3 1.3 1.3 1.2
Tot-N (%) 0.13 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1
Tot-P (g kg1 dw) 0.74 0.7 0.8 0.7 0.7 0.7 0.8 0.8 0.7
Tot-S (g kg1 dw) 0.20 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2
P-AL (mg 100 g1 dw) 4.5 4.0a 4.8bc 4.4ab 4.5 4.0a 5.0c 4.4 4.0a
P-Olsen (mg 100 g1 dw) 2.1 2.0 2.2 2.1 2.1 2.0 2.2 2.1 2.0
K-AL (mg 100 g1 dw) 19.8b 19.3 19.5 19.8b 20.6b 18.4a 20.3b 20.1b 18.4a
Cu (mg kg1 dw) 29.0 28.2 28.5 28.6 28.6 26.4 28.1 29.2 26.6
Zn (mg kg1 dw) 101.1 98.2 99.2 99.4 98.8 93.4 97.9 101.3 93.8
Cd (mg kg1 dw) 0.26 0.3 0.3 0.3 0.3 0.2 0.3 0.3 0.2
Cr (mg kg1 dw) 37.8 36.5 37.6 37.2 36.7 35.2 36.5 37.7 35.8
Ni (mg kg1 dw) 23.1 22.6 23.4 23.1 23.0 20.8 22.5 23.6 21.5
Pb (mg kg1 dw) 20.5 20.2 20.3 20.3 20.4 19.8 19.7 21.0 19.7
B-resp (lg CO2-C g1 dw h1) 0.23 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2
SIR (lg CO2-C g1 dw h1) 3.4 3.5b 3.4 3.4 3.4 3.2 3.3 3.6b 3.1a
lSIR (min1) 0.16 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2
Non-growers (lg CO2-C g1 dw h1) 2.9 3.0 3.0 3.0 3.0 2.8a 3.0 3.2b 2.8a
Growers (lg CO2-C g1 dw h1) 0.45bc 0.5c 0.4 0.5bc 0.4 0.5bc 0.3ab 0.4 0.3a
PDA (ng N2O-N g1 dw min1) 9.9 9.3 9.6 9.8 8.7 9.0 9.5 9.2 8.7
lPDA (min1) 0.25 0.3 0.3 0.3 0.3 0.2a 0.3 0.3b 0.2
PAO (ng NO2-N g1 dw min1) 4.2 4.1 4.0 4.1 3.9 3.6a 4.3 4.4b 4.0
Alk-P (lmol PNP g1 dw h1) 1.6 1.6 1.6 1.6 1.5 1.5 1.6 1.7 1.5
1
N-min (lg NHþ 4 -N g dw 10 d1) 18.9 18.6 19.0 18.7 20.5c 18.2ab 19.1 19.3bc 17.2a
1 1
Yield (kg ha year ) 2532.5bc 2666.7bc 2699.4bc 2704.7bc 2597.0bc 2798.9c 1946.8ab 2272.5 1593.4a
Means calculated as mean values over 4 years (n = 16). For abbreviations, see Tables 1–3. Statistically significant differences (Tukey p = 0.10) are indicated
by different letters.

Nitrogen mineralization was increased in BR100 and
cow manure treatments. These results are in agreement
with Tiwari et al. (2000) who found that application of bio-
gas slurry resulted in a proliferation of the microbial
population and increased nitrogen mineralization.
Together with soil respiration, potentially mineralizable
N is the most commonly suggested biological soil indicator
for assessing soil quality (Wienhold et al., 2004). The rea-
son for its usefulness is two-fold: (1) it may serve as a gen-
eral probe of the soil ecosystem status since most
microorganisms have at least some capacity to mineralize
N, and (2) it gives an estimate of the N deliverability to
the plants. The observed increase in nitrogen mineralization
indicates that biogas residues are beneficial to soil quality.
In contrast to organic-carbon-dependent respiration and
denitrification, autotrophic nitrification bacteria, also
being lithotrophic, are more dependent on pH and ammo-
nia availability since they use ammonia as their source of
energy. The nitrification process is vulnerable and is per-
formed only by highly specialized organisms (Johansson
et al., 1999), whose activity is enhanced by ammonia. It
was therefore expected that the ammonium-rich biogas res-
idue should increase PAO, compared to the nitrate-rich
NPS. The data indicated such an increase in PAO, but
the difference was not statistically significant.
4.3. Indicators of soil quality
Many of the existing methods used to evaluate treat-
ment effects on soil quality focus mostly on soil chemical
properties that change slowly with time, such as C, N
and P content, which limits their usefulness for detecting
short-term changes in soil quality. Measurements of the
status and activity of specific microbial communities con-
tributing to soil processes have the potential to provide
rapid and sensitive means of characterizing soil quality.
In this experiment the simple crop rotation and the homog-
enous experimental site chosen for the study made it possi-
ble to detect early responses of soil microorganisms to the
differences in composition of the organic wastes. Microbial
biomass (Doran and Parkin, 1994) and enzyme activity
(Badiane et al., 2001) have been proposed to indicate soil
quality. In this experiment the increase in microbial bio-
mass measured as SIR and the increase in several enzyme
activities suggests that the microorganisms responded rela-
tively fast to the content of ammonium and easy degrad-
able carbon.
5. Conclusions
Although the experimental period included 4 years of
nitrogen and organic matter supply of very different quali-
ties, surprisingly few significant differences in chemical and
microbiological properties could be observed. The most
prominent effect was that biogas residues increased the
substrate induced respiration, the proportion of active
microorganism, and the nitrogen mineralization, as well
as potential ammonia oxidation. In all, studying microbial
parameters seems to be a better choice than chemical
 M. Odlare et al. / Waste Management 28 (2008) 1246–1253
parameters when short-term effects of organic wastes are to
be evaluated.
In addition, the results can help to assess the relative
benefits of the various organic residues tested. No negative
effects on the soil microbial properties studied could be
observed by any of the applied organic wastes products
and they all seemed to be equally good or better as fertiliz-
ers as compared to cow manure, pig slurry and NPS. Bio-
gas residue contained larger concentrations of mineral
nitrogen and easy degradable carbon, and was therefore
more efficient than the other organic wastes in terms of
supplying plant-available nitrogen and promoting the soil
biological activity. Sewage sludge supplied the soil with
potassium and phosphorous, but the mineral nitrogen con-
tent did not match that found in the biogas residue. Com-
post contained low concentrations of mineral nitrogen and
the microbial mineralization rate seemed to be too slow to
meet the nitrogen requirements of agricultural plants.
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