POLISH
JOURNAL
VOL. XXXVIII/2
OF
2005
SOIL
SCIENCE
PL ISSN 0079-2985
Soil Biology
STEFANIA JEZIERSKA-TYS, MAGDALENA FR¥C*
THE EFFECT OF FERTILIZATION WITH SEWAGE SLUDGE
FROM A DAIRY PLANT AND WITH RAPE STRAW ON THE
POPULATION NUMBERS OF SELECTED MICROORGANISMS
AND RESPIRATION ACTIVITY OF BROWN SOIL
Received February 22, 2006
Abstract. The research was carried out in a pot experiment. The effect of dairy sewage sludge and
rape straw on the numbers of bacteria and fungi and on the respiration activity was examined. The pot
experiment included three treatments: 1 – control soil, 2 – soil + dairy sewage sludge, 3 – soil + dairy
sewage sludge + rape straw. As a result of laboratory investigations, the addition of sewage sludge
from a dairy plant to soil was found to stimulate the total number of bacteria, actinomycetes, and
fungi and the number of cellulolytic bacteria, as well as to contribute to the increase in the respiration
activity of soil.
The microbiological activity and fertility of soil are closely interrelated, as it is
the biomass that directly participates in the mineralization of major organic
compounds [22]. Fertilizers, residues of crop and other organic matter are highly
significant to the maintenance of soil quality in respect of yield increase [2].
Sewage sludge contains a considerable amounts of nutritive value especially due to
a high nitrogen content [3, 6, 12, 29]. Sewage sludge from dairy plant has a low
C:N ratio [5, 6], therefore it can be applied for the fertilization of soil together with
straw [5] which exhibits a high C:N ratio. Straw affects the improvement of soil
properties, especially organic carbon accumulation in the soil [17]. Due to the high
C:N ratio in straw, its administration into the soil enables immobilization of
mineral nitrogen by microorganisms which become a competitor to field crops [7,
19]. In order to reduce this phenomenon, additional fertilization with nitrogen
originating from mineral or organic fertilizers is applied [7-9].
*Asst. Prof. S. Jezierska-Tys, DSc.; M. Fr¹c, MSc.; Department of Agricultural Microbiology,
University of Agriculture, St. Leszczyñskiego 7, 20-069 Lublin, Poland.
146
S. JEZIERSKA-TYS, M. FR¥C
The experiment aimed to determine the effect of sewage sludge from a dairy
plant and straw on the respiration activity and numbers of bacteria and fungi in
brown soil.
MATERIALS AND METHODS
A pot experiment was established with brown soil formed from silt-loam with
8% sand fraction, 47% slit fraction and 45% clay and slit fraction. The soil had
slightly acid reaction (pH 6.4) and contained 0.16% of nitrogen and 1.8%
C-organic. Sewage sludge used in the experiment originated from the District
Dairy Cooperative in Krasnystaw. The dairy sewage sludge was hygenised with
ash of lignite and contained 300 g kg-1 of dry matter and 33.2 g kg-1 of nitrogen in
dry matter, as well as reaction approximatly neutral.
The pot experiment design included three treatments: 1 – control soil, without
fertilization, 2 – soil + sludge 7.3 g kg-1, 3 – soil + sludge 6.2 g kg-1 + rape straw 1.7
g kg-1. Soil was watered to 60% of total water capacity and incubated at a
temperature of 20°C, with moisture content being kept constant over the entire
experimental period. Analyses were carried out in three replications, in: day 7 - I
term, day 14 – II term, day 30 – III term, day 60 – IV term, and day 90 of the
experiment – V term, and involved counts of: the number of cellulolytic bacteria on
liquid culture medium that consists of (g dm-3) KNO3 1, K2HPO4 0.5, MgSO4 0.2,
NaCl 0.2, MnSO4 trace, FeSO4 trace, CaCO3 5, distilled water 1000 cm3, the
numbers of bacteria and actinomycetes on culture medium which consists of (g
dm-3) soil extract 350 cm3, K2HPO4 0.5, glucose 1, agar 15 and distilled water 650
cm3, the number of fungi on Martin medium [18], as well as determinations of the
respiration activity with the Rühling and Tyler’s method [26], and pH – with
potentiometric method. The number of cellulolytic bacteria was determined with
the test-tube method, whereas the most probable number (MPN) of bacteria was
read out from statistical tables of Mc Crady [25] for three parallel repetitions. Total
numbers of bacteria, actinomycetes and fungi were assayed with the method of
peur plates. The results obtained were elaborated statistically with the analysis of
variance using Statistica 6.0 software.
RESULTS AND DISCUSSION
Due to a lack of reference data on the effect of dairy plant-originated sewage
sludge on the microbiological activity of soil, an attempt was made to discuss the
results obtained for that sludge with those reported for sludge originating from
municipal and industrial sewage treatment plants.
Changes in the respiration activity of soil are presented in Fig. 1. An analysis of
the results obtained indicates that at the beginning of the experiment (terms I and
EFFECT OF FERTILIZATION WITH SEWAGE SLUDGE FROM A DAIRY PLANT
147
180
mg CO 2 · kg
-1
·d
-1
160
140
120
100
80
60
40
20
0
I
1
II
2
3
III
IV
V
Dates of analyses
Fig. 1. Soil respiration activity as influenced by sewage sludge and straw application LSD0.05 for: D =
25.07, T =16.52, DT = 55.17. D - dates of analyses, T - treatments. Explanations: 1 - control soil, no
sludge; 2 - soil + dairy sewage sludge; 3 - soil + dairy sewage sludge + rape straw.
II) the sewage sludge evoked a significant increase in the respiration activity of soil
both in the treatment with sludge alone and in the soil fertilized with sludge and
straw. It should be pointed out that in the first term the activity was significantly
higher in the treatment with the sludge and straw, whereas in the second term it was
insignificantly higher in the soil fertilized with the sludge alone. It is also worth
emphasizing that beginning from the third term till the end of the experiment the
respiration activity was decreasing, and was lower than in the control soil. Similar
results were obtained by Pascual et al. [23]. Their investigations also demonstrated
a decrease in the respiration activity over the experimental period. As in our
research, the above-mentioned authors observed that sewage sludge exerted a
stimulating effect on soil respiration in the initial phase of the experiment. Then
they observed the decrease of the respiration activity below values in the control
soil. Investigations of Moreno et al. [20], Tasatar and Haktanir [27] as well as
Atman and Arcak [1] have demonstrated that a sewage sludge dose similar to that
applied in our study had a beneficial effect on the respiration activity of soil. In
addition, Tasatar and Haktanir [27] reported the greatest stimulating effect of the
sludge in the initial phase of the experiment and a decrease in the activity in the
course of the study. On the contrary, Emmerling et al. [4] showed that the
respiration activity of soil was stimulated to the greatest extent by sludge
supplemented with coal fly ash. In a field experiment, Wielgosz [28] also reported
on the highest amount of CO2 released at the initial stage of analyses. Quemada and
Menacho [24] showed that one year after sewage sludge application the respiration
activity of soil was higher in the soil fertilized with sewage sludge than in the
control one. Research carried out by Johansson et al. [10] demonstrated that the
148
S. JEZIERSKA-TYS, M. FR¥C
addition of sewage sludge did not affect soil respiration, still it should be
emphasized that in this study the sludge was applied at a dose of 12 t ha-1, i.e. twice
as low as in our experiment.
Figure 2 presents the number of cellulolytic bacteria in soil fertilized with dairy
plant sewage sludge and rape straw. In the initial phase of the experiment, the
number of those bacteria was observed to decrease and become lower than that
reported for the control soil. It is worth emphasizing that after 14 and 30 days of soil
incubation, the number of cellulolytic bacteria was stimulated only in the soil
applied with the sludge and straw, whereas in the treatment with sludge alone – it
was lower than in the control. After 60 and 90 days of incubation, a significantly
higher number of those bacteria was observed only in the soil fertilized with the
sewage sludge alone.
160
140
100
MPN · 10
4
120
80
60
40
20
0
I
1
2
3
II
III
IV
V
Dates of analyses
Fig. 2. Number of cellulolityc bacteria in soil as influenced by sewage sludge and straw application.
Explanations on Fig. 1.
Kucharski et al. [15] observed a favourable effect of straw addition to soil on
the proliferation of cellulolytic bacteria, whereas investigations of Kobus et al.
[13] demonstrated that the number of those bacteria did not increase upon soil
fertilization with sewage sludge. Research carried out by Nowak et al. [21] did not
demonstrate any changes in the numbers of cellulolytic bacteria neither in the soil
fertilized with composted sewage sludge nor in the soil enriched with composted
sewage sludge and straw. In contrast, they reported that the number of those
bacteria in the soil was decreasing along with the proceeding time of incubation,
which was also observed in our experiment in the soil fertilized with sewage sludge
and straw. This can be explained by gradual depletion of nutrients for that group of
bacteria.
The sewage sludge and straw were observed to insignificantly stimulate the
number of fungi (Fig. 3) after 14, 30, and 60 days of incubation, whereas after 7 and
EFFECT OF FERTILIZATION WITH SEWAGE SLUDGE FROM A DAIRY PLANT
149
25
CFU · 10 7
20
15
10
5
0
I
1
2
II
III
3
IV
V
Dates of analyses
Fig. 3. Total number of fungi in soil as influenced by sewage sludge and straw application. LSD0.05
for: D = 2.93 107, T = not significant, DT = 6.46 107. D – dates of analyses, T - treatments.
Explanations on Fig. 1.
90 days of incubation (at the beginning and at the end of the experiment) the
number of those microorganisms was lower than in the control soil. It was likely to
result from depletion of substrate for that group of bacteria. In addition, significant
changes in fungi number were observed in particular periods of analyses.
Investigations of Kacprzak and Stañczyk-Mazanek [11] demonstrated that the
administration of sewage sludge caused the fungi number to increase from the
beginning of the experiment up to 6 months of incubation, and then to approximate
the number of fungi reported for the control soil.
Results of the effect of sewage sludge from a dairy plant on the numbers of
bacteria and actinomycetes is presented in Fig. 4. The results of this research
indicated a significant stimulation of the numbers of bacteria and actinomycetes
was observed in both treatments with the sludge. Except for the object fertilized
45
40
CFU · 10
9
35
30
25
20
15
10
5
0
I
1
2
3
II
III
IV
V
Dates of analyses
Fig. 4. Total number of bacteria and actinomycetes in soil as influenced by sewage sludge and straw
applicationLSD0.05 for: D = 4.10 109, T = 3.49 109, DT = 9.36 109. D – dates of analyses, T - treatments.
Explanations on Fig. 1.
150
S. JEZIERSKA-TYS, M. FR¥C
with the sludge alone where after 60 days of incubation the numbers of bacteria and
actinomycetes dropped to the values reported for the control soil. It should be
pointed out that the numbers of those bacteria and actinomycetes were most
significantly stimulated in the soil fertilized with straw, which was likely to result
from the change in the C:N ratio.
Nowak et al. [21] observed a similar tendency when fertilizing soil with
composted sewage sludge and composted sewage sludge with the addition of
straw, still the numbers of both bacteria and fungi were more strongly stimulated in
the soil enriched with compost and straw than in the soil with the sludge application
alone. Kobus et al. [13] also reported that the addition of sewage sludge to soil
considerably increased the numbers of bacteria, actinomycetes and fungi.
Kucharski and Niklewska [14] proved that fertilization with straw had a beneficial
effect on the proliferation of the bacterial groups examined. Those results have also
been confirmed in our experiment. A number of authors [4, 10, 16, 23] have
demonstrated the stimulating effect of sewage sludge on soil microflora.
CONCLUSIONS
1. Dairy sewage sludge as well as dairy sewage sludge with the addition of
straw were found to significantly stimulate the respiration activity of soil.
2. Introduction of sewage sludge from a dairy plant to the soil caused a
significant increase in the numbers of bacteria and actinomycetes.
3. Straw administered to the soil together with the dairy sewage sludge
significantly stimulated the proliferation of bacteria and actinomycetes as well as
cellulolytic bacteria, still it exerted the most beneficial effect on bacteria and
actinomycetes.
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WP£YW NAWO¯ENIA OSADEM ŒCIEKOWYM Z MLECZARNI I S£OM¥
RZEPAKOW¥ NA LICZEBNOŒÆ WYBRANYCH MIKROORGANIZMÓW
ORAZ AKTYWNOŒÆ RESPIRACYJN¥ GLEBY BRUNATNEJ
W doœwiadczeniu wazonowym badano wp³yw osadu œciekowego z mleczarni i s³omy
rzepakowej na liczebnoœæ bakterii i grzybów oraz aktywnoœæ respiracyjn¹ gleby brunatnej. Schemat
doœwiadczenia obejmowa³ trzy obiekty doœwiadczalne: 1 – gleba kontrolna, 2 – gleba + osad œcieków
mleczarskich, 3 – gleba + osad œcieków mleczarskich + s³oma rzepakowa. W wyniku badañ
laboratoryjnych stwierdzono, ¿e wprowadzenie do gleby osadu œciekowego z mleczarni wp³ywa³o
dodatnio na liczebnoœæ bakterii i promieniowców, grzybów oraz bakterii celulolitycznych, a tak¿e
przyczynia³o siê do zwiêkszenia aktywnoœci respiracyjnej gleby.