Waste Management 26 (2006) 12531258 www.elsevier.

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Eects of composted sewage sludge on microbial biomass, activity and pine seedlings in nursery forest
S.Yu. Selivanovskaya *, V.Z. Latypova
Department of Applied Ecology, Ecological Faculty, Kazan State University, Kremlyevskaya, 18, Kazan 420008, Russia Accepted 6 September 2005 Available online 22 November 2005

Abstract The investigation was carried out in a 2 year experiment to evaluate the benets and hazards of the use of composted sewage sludge as a restoration agent for the soil of the nursery forest intended for growing Pinus sylvestris seedlings. The grey forest soil (Haplic Greyzem) was amended with compost at the 25, 50, 75, 100, 150 and 175 t ha1 application rates on a dry matter basis. The organic matter content increased with the increase in sludge amendment as well as the metal content. However, the concentrations of individual metals were below the current limits established for Russia and European countries. Sludge amendments enhanced the germination and decreased the mortality of the seedlings. The eects were more obvious for the soil with the highest sludge treatment. The benecial eects on the biomass of seedlings and the height of the shoots as well as on the length of the roots of the pine seedlings were greater in plots with the highest rates of composted sludge. The application of composted sludge to soil was followed by an increase in microbial biomass and to a lesser extent in basal respiration. In the absence of any detrimental eect on microorganisms, this study lends support to using composted sewage sludge as the organo-mineral fertilizer for the soil of nursery forest. 2005 Elsevier Ltd. All rights reserved.

1. Introduction Large amounts of sewage sludge have been accumulated and expected to be generated in Russia due to the increasing volume of wastewater treated. Current trends in European waste policy aim at reducing biodegradable wastes in landll sites (Debosz et al., 2002). Another disposal option, i.e. land application, is commonly used in most municipalities. Land application of sewage sludge provides not only a means for sludge disposal but can also improve soil fertility and the physical properties of the soils, causing increase in crop yield (Peles et al., 1996; Ramachandran and DSoura, 1998; Gardiner et al., 1995; Jorba and Andres, 2000). The agricultural use of sewage sludge is mandated by Russian law (SanPiN No 2.1.7.573-96, 1997). However, only 1% of the sewage sludge generated is used in agriculture. Unfortunately, the eects of sewage sludge on soil quality
Corresponding author. Tel.: +7 8432 930782; fax: +7 8432 920554. E-mail address: Svetlana.Selivanovskaya@ksu.ru (S.Yu. Selivanovskaya). 0956-053X/$ - see front matter 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.wasman.2005.09.018
*

are dual. On the one hand, sludge applications to soil increases the content of organic matter and plant nutrients; on the other hand, the presence of pathogens and heavy metals limits the general use of sewage sludge in agriculture, especially that derived from both industrial and domestic sources. The safe utilization of the sewage sludge has to be ensured by its preliminary treatment. Composting might be a possible and practical means for reducing metal contents and the amount of pathogens in the sewage sludge (Page, 1996; Selivanovskaya et al., 2001; Sanchez-Monedero et al., 2004). Furthermore, as previously shown the composted sewage sludge causes the minimal alteration of the pre-existing equilibrium of soil microbial communities and a prolonged positive eect on soil properties in comparison with the anaerobically digested or untreated sludge (Selivanovskaya et al., 2001; Ros et al., 2003). Whereas most of the works have focused on the eect of the sewage sludge on soil fertility and crop quality in agriculture, the eect of the composted sewage sludge used as a fertilizer in forestry, namely in the nursery forest is less known. Meanwhile, the experience of the sewage sludge

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application to the soil of the nursery forest is of special importance because the Council Directive 86/278/EEC on the use of sewage sludge in agriculture is currently under revision when regulating the use of sewage sludge not only in agriculture but also elsewhere, e.g., in forestry, green areas and land reclamation (Sanchez-Monedero et al., 2004). The main objective of this study was to quantify the eects of the dierent rates of composted sewage sludge on the microbial activities and microbial biomass of degraded soil of the nursery forest in the course of two years. The evolution of edaphic conditions as well as some properties of the pine seedlings cultivated on this soil are also presented. 2. Materials and methods The study was carried out at the forest nursery Matyushenski, Tatarstan, Russia. Quadruplicate eld plots were established on the grey forest soils (Haplic Greyzem) in a randomized block design. The plots of 2.5 3.5 m were sawed with Pinus sylvestris. The composted sewage sludge was prepared from untreated sludge, peat and the mixture of the wood shaving and sawdust taken in the ratio 1:1:1 on a dry solid basis. The materials were fermented for 6 weeks followed by 7 months maturation. The compost from sewage sludge contained 22.1% organic C, 1.7% total N, 546 74 mg Zn kg1, 12 2 mg Cd kg1, 77 14 mg Ni kg1, 167 42 mg Cr kg1, 312 45 mg Cu kg1, 166 37 mg Pb kg1, 669 133 mg Mn kg1. The composted sewage sludge was added at the rates of 25, 50, 75, 100, 150 and 175 t ha1 on a dry matter basis. There was no sludge addition to the control plots. All the treatments were applied before the pine seeds were sown. Five soil samples were collected from each of four replicates at depth of 00.1 m. The samples were pooled. The mass of pooled samples of each replicate was about 0.5 kg. All visible plant debris and soil fauna were removed manually. The soils were then sieved (2 mm), mixed to ensure homogeneity and stored at 4 C prior to analysis. The rst soil sampling was carried out on 7 May 2001. The plants

(50 in each plot) were harvested by hand after two years of growth and then weighed. The growth and biomass parameters were calculated for a single plant. The plant samples were oven dried at 80 C for 48 h, ground with an agate mill prior to analysis. All the samples were used for the heavy metal analysis within 24 h after sampling. Organic C was determined by wet oxidation with 0.167 M K2Cr2O7 and by subsequent titration with 0.1M (NH4)2Fe(SO4)2 6 H2O. The total content of acid extractable heavy metals was determined in 1 g samples of soil, sewage sludge and ashes of the plants using wet oxidation with the concentrated HNO3 and 3% H2O2. The metals were determined by atomic absorption spectroscopy with the Aanalyst-300 (PerkinElmer). Soil microbial biomass (Cmicr) was determined by fumigation of the samples with C2H5OH-free CHCl3 and extraction with 0.5 M K2SO4. The extracted C content was determined by bichromate oxidation in accordance with ISO 14240-2 (1997). Basal respiration (CO2 evolution with no substrate addition) was determined in 10 g of soil adjusted to 60% water holding capacity and incubated at 20 C. CO2 was trapped in 1 M NaOH and measured by titration with 0.1 M HCl after the addition of BaCl2 and indicator solution (Schinner, 1995). All the results were expressed on an air-dry soil basis and based on ve replicates. Statistical analyses were performed using the analysis of variance (ANOVA) for a randomized complete block design with a = 0.05. The means were compared using Fishers Protected Least Signicant Dierence at a = 0.05. All the tables presented include standard errors of the data. 3. Results and discussion 3.1. Eects on Corg and total metal content in grey forest soil The Corg levels obtained at the end of the second year of investigation are compiled in Table 1. The addition of increasing amounts of composted sewage sludge increased

Table 1 Organic matter (%) and metal contents (mg kg1) of grey forest soil amended with compost of sewage sludge at the rates of 25175 t ha1 and unamended (control) soil Variant Control 25 50 75 100 150 175 CEC limitsa Limits for Russia
a b b

Corg (%) 0.7 0.1 1.0 0.2 1.2 0.2 1.4 0.3 1.5 0.3 1.9 0.4 2.1 0.4

Zn 15.1 3.1 10.3 1.9 19.6 1.5 17.5 2.1 18.7 2.4 26.2 7.5 28.1 5.3 150300 55220

Cu 31.4 19.6 73.8 11.2 53.7 8.1 77.3 22.4 54.6 7.4 58.5 9.2 62.3 14.7 50140 33132

Cd 0.4 0.2 1.4 0.3 1.1 0.2 1.1 0.2 1.7 0.4 1.6 0.1 1.9 0.4 13 0.52

Ni 24.8 5.4 24.0 4.8 27.5 6.2 28.2 6.9 32.1 7.6 27.1 6.1 38.0 8.7 3075 2080

Mn 760.8 344.1 1198.0 4558.5 1346.9 289.4 1707.6 389.6 1360.4 255.7 1039.4 207.3 1493.5 288.2

Cr 47.1 14.8 47.6 11.2 56.1 14.2 104.3 19.8 113.9 22.3 69.8 11.8 96.3 15.5 100150 150

Pb 9.9 2.1 16.5 3.4 8.9 1.4 14.4 3.2 15.0 3.8 8.1 1.4 17.8 2.9 50300 32130

(McGrath et al., 1995). SanPiN No 2.1.7.573-96, 1997.

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Table 2 Changes in microbial biomass C (mg Cmicr g1) in samples of unamended grey forest soil (control) and in samples after application of composted sludge Time, day after compost application Soils amended with compost at the rates 25 1 32 66 124 360 391 452 0.57 0.09 0.38 0.06 0.36 0.06 0.19 0.03 0.65 0.11 0.25 0.04 0.26 0.04 50 0.86 0.12 0.80 0.11 0.29 0.04 0.15 0.02 0.31 0.04 0.60 0.08 0.28 0.03 75 0.97 0.20 0.82 0.17 0.31 0.06 0.181 0.04 0.32 0.06 0.5 0.10 0.55 0.15 100 0.74 0.12 0.56 0.09 0.62 0.10 0.36 0.06 0.55 0.09 1.26 0.21 0.6 0.10 150 0.83 0.19 0.62 0.14 0.67 0.16 0.46 0.11 0.80 0.19 0.81 0.19 0.95 0.22 175 1.84 0.46 0.98 0.24 0.88 0.22 0.48 0.12 0.70 0.17 0.85 0.21 0.85 0.22 0.44 0.09 0.11 0.02 0.20 0.04 0.12 0.02 0.49 0.10 0.35 0.07 0.31 0.06 Control soil

Corg from 0.7% for the control (unamended) soil to 2.1% for the soil amended with the highest rate (175 t ha1) of compost due to the high organic matter in the compost. The metal content of the soil samples was determined seven times during the two years of investigation. The mean metal concentrations for the entire study period are presented in Table 1. The results indicate an increasing trend of soil metal content for all the metals with the increasing rate of the compost applied. Signicant dierences (P < 0.05) were observed between the means of metal content in the treated and the control soil. However, even in the compost amended plots, the concentrations of individual metals were found to be below the current limits established for Russia and European countries (McGrath et al., 1995; SanPiN No 2.1.7.573-96, 1997). 3.2. Eects on soil microbial biomass and activity Soil microorganisms play an essential role in the decomposition of soil organic matter. Any disturbance to the soil ecosystem can disrupt the activity and hence the nutrient cycling. Microbial parameters are considered to be sensitive indicators of an ecology stress suered by the soil and its recovery, because microbial activity has a direct inuence on the stability and fertility of ecosystems (Chander and Brookes, 1991, 1993; Ros et al., 2003). Microbial parameters are very responsive to small changes in the soil. Microbial biomass is one of the most commonly used parameters. As shown in Table 2, compost application generally increased the soil microbial biomass carbon (Cmicr). This increase was greater for the higher rates of amendment. On the rst day after the compost application, the maximum values of this parameter were found to be 0.57, 0.86, 0.97, 0.74, 0.83, and 1.8 mg Cmicr g1 for the soils amended with the composted sewage sludge at the rates of 25, 50, 75, 100, 150 and 175 Mg ha1, respectively. The appropriate value for the control soil was found to be 0.44 mg Cmicr g1. This can be attributed either to the incorporation of the easily biodegradable organic matter, which stimulates authochtonous microbial activities in the soil or to a new microbial biomass already grown in the composted material added to the soil. A similar inuence of sewage sludge on the biomass has been reported by other authors (Pascual et al., 1997; Moreno et al., 1999; Garcia-Gil et al., 2000; SanchezMonedero et al., 2004). With time, Cmicr decreased in all the

treatments. However, in the soil amended with high rates of the composted sludge (100, 150 and 175 Mg ha1), the biomass was found signicantly (P < 0.05) greater by a factor from 2 to 3.1 than that determined in the control soil throughout the measured period of two years. The metal content in the soil under dierent treatments showed an increase in their levels with the addition of compost (Table 1). However, the minimal metal concentrations that decrease the microbial biomass in the soils with a similar organic matter content and pH are higher than the metal contents determined in this study (McGrath et al., 1995). This is probably the reason why no decrease in the soil microbial biomass was found. Besides, the positive eects of the organic matter added serve to counterbalance the negative eect caused by the metals present. Soil microbial respiration is expressed as the CO2C released per day and gram of soil and is considered a useful indicator for measuring soil microbial activity (Anderson and Domsch, 1990). The initial basal respiration values in all the amended soils were lower than those expected from the Cmicr levels (Table 3). In the course of 3060 days, the CO2C evolution was increased in all the soils. Compost application had no clear-cut eect on basal respiration. Subsequently, the basal respiration decreased during the rst year probably due to the substrate depletion and decrease in Cmicr. In relation to microbial respiration, it has been argued that high metal concentration decreases the CO2 evolution whereas moderate contamination leads to higher respiration rates (Khan and Scullion, 2000). The negligible eects of metals on soil respiration observed in this study were not surprising. In fact, the negative eects of metals have been reported, but only at a much higher concentration (McGrath et al., 1995). The inhibition in the initial basal respiration can be most likely attributed to phytotoxic organic metabolites present in compost (Pascual et al., 1997; Moreno et al., 1999). However, after a month toxic compounds disappeared which led to an increase in the basal respiration. The relationship between soil respiration and microbial biomass can be expressed using the parameter of the metabolic quotients qCO2 calculated as mg CO2C 24 h1 mg biomass C. Organic amendment of the soil resulted in an increase in qCO2 by the 120 days of investigation which appeared particularly high in the case of the low rates of

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Table 3 Changes in basal respiration (mg CO2C g1 24 h1) in samples of unamended grey forest soil (control) and in samples after application of composted sludge Time, day after compost application Soils amended with compost at the rates 25 1 32 66 124 360 391 452 0.16 0.03 0.17 0.03 0.14 0.02 0.20 0.04 0.13 0.02 0.03 0.007 0.09 0.02 50 0.08 0.03 0.16 0.05 0.26 0.08 0.20 0.06 0.16 0.05 0.12 0.04 0.17 0.06 75 0.05 0.02 0.32 0.09 0.27 0.07 0.31 0.09 0.17 0.04 0.07 0.02 0.16 0.04 100 0.19 0.08 0.45 0.18 0.27 0.11 0.27 0.11 0.04 0.02 0.09 0.03 0.12 0.05 150 0.20 0.07 0.23 0.08 0.21 0.08 0.30 0.11 0.05 0.02 0.11 0.04 0.13 0.05 175 0.22 0.06 0.29 0.08 0.15 0.04 0.32 0.09 0.02 0.007 0.12 0.03 0.12 0.03 0.18 0.04 0.15 0.04 0.08 0.02 0.15 0.03 0.02 0.005 0.07 0.02 0.12 0.02 Control soil

Table 4 Changes in metabolic quotient (qCO2) (mg CO2C mg1 24 h1) in samples of unamended grey forest soil (control) and in samples after application of untreated, anaerobically digested and composted sludge Time, day after compost application Soils amended with compost at the rates 25 1 32 66 124 360 391 452 0.28 0.44 0.39 1.04 0.20 0.11 0.36 50 0.09 0.20 0.86 1.35 0.52 0.20 0.59 75 0.05 0.39 0.87 1.70 0.53 0.14 0.28 100 0.26 0.81 0.43 0.76 0.08 0.07 0.20 150 0.24 0.36 0.31 0.64 0.06 0.14 0.14 175 0.12 0.30 0.17 0.66 0.03 0.14 0.14 0.42 1.40 0.41 1.26 0.05 0.20 0.41 Control soil

the compost (2575 t ha1) (Table 4). However, even in these cases the qCO2 values did not dier signicantly (P < 0.05) from those of the control soil. This increase can be attributed to the decrease in Cmicr (Table 2) which was observed in this period in all the soils. During the second year, the qCO2 values of all the amended soils were similar to those of the control soil. As far as qCO2 is concerned, it is suggested to be used as a marker of the environmental disturbance or stress of the microbial population (Anderson and Domsch, 1993). The absence of signicant eects reected in the qCO2 values indicates the least alteration of the ecosystem. Moreover, the dynamics of the qCO2 suggests the microorganisms undergo the disturbance caused by the addition of organic matter rather than stress due to the presence of metals. In fact, minimum metal concentrations that decrease microbial biomass (one of the parameters used for qCO2 calculation) cited by McGrath et al. (1995) are higher than the metal contents determined in this study for the soils with a similar organic matter and pH. 3.3. Eects of composted sewage sludge on germination of pine seedlings, their behavior and metal content During the rst year, the germination of the P. sylvestris seeds and the seedling mortality were estimated. The germination of the seeds was aected by treatments. Table 5 indicates the increasing seed germinations with the increasing rates of compost application. Generally, all the treatments demonstrated that pine seeds germination is signicantly (P < 0.05) greater by a factor from 1.3 to 1.8 than that determined in the control soil. The dierence in the germi-

Table 5 Percentage of germination of the Pinus sylvestris seeds depending on the treatments Variant Control 25 50 75 100 150 175 Germination (%) 45 57 61 64 66 71 82

nation levels was particularly noticeable in all the treatments from the seventh day of the experiment. Maximal percentage of germination was attained towards the seventeenth day after sowing. For all the compost rates, seedling mortality during the rst two months was smaller in the compost treated than in the control soil. The present ndings are in agreement with the reports of Jorba and Andres (2000) who found that sewage sludge could serve as a good organic fertilizer to maximize plant germination. Two years after planting, the mass of the plants, the height of the shoots and the length of the roots of the pine seedlings were measured. Compost treatment eects were uniformly signicant for the height of the shoots and the length of the roots that had markedly increased with the compost loading (Fig. 1). The addition of the compost in the rates from 25 to 175 t ha1 increased the height of the shoots from 14.9 to 19.8 cm and the length of the roots from 18.8 to 20.6 cm. The appropriate value for the seedlings of the control soil was found to be 8.7 and 15.1 cm for the shoots and roots, respectively.

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shoots roots

1257
shoots roots

5 4

25

biomass, g dry matter

20

3 1

size,cm

15

10

0 1 2 3 4 variant 5 6 7

0 1 2 3 4 5 6 7

Variant
Fig. 1. Height of the shoots and length of the roots of the pine seedlings depending on the treatments. 1 control, 2 25 t ha1, 3 50 t ha1, 4 75 t ha1, 5 100 t ha1, 6 150 t ha1, 7 175 t ha1.

Fig. 2. Biomass of shoots and roots per seedlings depending on the treatments. 1 control, 2 25 t ha1, 3 50 t ha1, 4 75 t ha1, 5 100 t ha1, 6 150 t ha1, 7 175 t ha1.

The total dry weight yields of Pinus sylvestris in soils amended with various rates of the compost are shown in Fig. 2. All the compost additions produced a signicantly higher yield of seedling shoots than the control soil. The yield of the seedling roots was not signicantly aected by the compost treatment. Table 6 shows the content of various metals in the shoots and roots of the plants. The metal contents did not correlate with the rates of the compost applied. The concentrations of each metal were similar in all the roots and shoots collected from the treated and untreated plots. Within all the plots amended with the compost, the highest metal concentrations were found in the roots of pine seedlings. In general, the Zn and Mn concentrations were higher in comparison with that of Cd and Cr. Probably, this is due to the specic properties of these elements.

Besides, high Zn and Mn concentrations suggest greater capacity of the adsorption of these elements by this particular species. The metal contents in tissues for all the treatments were below their toxicity levels. The Cu, Zn, Cr, Ni, Mn, Cd, Pb contents did not exceed the toxicity levels of 100, 400, 80, 100, 500, 80, 60 mg kg1, respectively (Smith, 1994; Miller et al., 1995; Wong et al., 2001). 4. Conclusion The study showed that the amendment of the sewage sludge compost aected several chemical and biological parameters. The inuence became mainly evident in the rst weeks after amendment. The organic matter content increased with the increase in sludge amendment as well as in metal content. However, the concentrations of individual metals were below the current limits established for Russia and European countries. No severe negative

Table 6 Metal contents (mg kg1 on dry matter) in the shoots and roots of pine seedlings after two years of the growth Variant Shoots j 25 50 75 100 150 175 Roots j 25 50 75 100 150 175 Cr 5.9 1.3 1.7 0.4 26.2 5.2 3.8 0.7 1.1 0.2 1.1 0.2 2.3 0.5 Cu 35.6 6.2 64.0 16.1 93.0 22.4 34.2 7.8 32.4 7.1 45.0 9.5 31.3 6.4 Mn 20.4 4.2 22.8 4.9 40.0 7.8 104.4 18.3 13.1 3.5 5.3 1.1 5.7 1.2 Ni 4.5 0.9 1.3 0.4 27.8 6.2 1.6 0.3 4.9 1.1 0.9 0.2 1.7 0.4 Zn 95.0 22.3 254.3 49.1 116.6 22.4 74.6 11.6 88.9 18.4 128.5 24.4 81.3 16.3 Pb 16.8 3.4 24.2 5.9 38.8 8.1 15.2 3.9 15.7 3.1 17.4 4.2 15.6 3.1 As 0.15 0.03 0.09 0.02 0.52 0.15 0.08 0.02 0.06 0.01 0.06 0.01 0.10 0.02 Cd 0.06 0.01 0.01 0.002 0.01 0.002 0.04 0.003 0.01 0.002 0.01 0.002 0.03 0.007

2.7 0.6 1.2 0.3 1.7 0.4 2.8 0.6 1.5 0.4 3.8 0.6 1.3 0.3

27.6 5.9 17.4 3.7 29.7 3.8 21.3 5.1 41.7 7.9 37.9 6.4 24.2 4.2

8.0 1.4 2.4 0.5 9.0 1.6 5.2 1.1 8.7 1.2 9.9 1.1 4.1 0.8

4.7 1.1 3.8 0.7 16.1 3.2 5.3 1.1 8.4 1.9 30.2 6.7 6.0 1.2

45.2 11.2 44.3 10.4 50.1 12.8 41.8 6.9 85.5 14.8 60.3 12.5 42.7 8.0

15.7 2.8 9.3 1.5 15.8 3.2 10.0 1.7 20.3 4.1 19.8 3.5 12.2 3.1

0.34 0.07 0.05 0.01 0.25 0.47 0.10 0.02 0.24 0.06 0.69 0.12 0.15 0.03

0.01 0.002 0.01 0.002 0.01 0.002 0.01 0.002 0.01 0.002 0.01 0.002 0.01 0.002

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S.Yu. Selivanovskaya, V.Z. Latypova / Waste Management 26 (2006) 12531258 Jorba, M., Andres, P., 2000. Eects of sewage sludge on the establishment of the herbaceous ground cover after soil restoration. J. Soil Water Conserv. 3, 322326. Khan, M., Scullion, J., 2000. Eect of soil on microbial responses to metal contamination. Environ. Pollut. 110, 115125. McGrath, S.P., Chaudri, A.M., Giller, K.E., 1995. Long-term eects of metals in sewage sludge on soils. microorganisms and plants. J. Ind. Microbiol. 14, 94104. Miller, R.W., Azzari, A.S., Gardiner, D.T., 1995. Heavy metals in crops as aected by soil types and sewage sludge rates. Commun. Soil Sci. Plant Anal. 29, 703711. Moreno, J.L., Hernandez, T., Garcia, C., 1999. Eects of cadmiumcontaminated sewage sludge compost on dynamics of organic matter and microbial activity in an arid soil. Biol. Fert. Soils 28, 230237. Page, A.L., 1996. Use of Reclaimed Water and Sludge in Food Crop Production. National Academy Press, Washington, DC. Pascual, J.A., Garcia, C., Hernandes, T., Ayuso, M., 1997. Changes in the microbial activity of the arid soil amended with urban organic wastes. Biol. Fert. Soils 24, 429434. Peles, J.D., Brewer, S.R., Barrett, G.W., 1996. Metal uptake by agricultural plant species Grown in sludge-amended soil following ecosystem restoration practices. Bull. Environ. Contam. Toxicol. 57, 917923. Ramachandran, V., DSoura, T.J., 1998. Plant uptake of cadmium, zinc, and manganese in soils amended with sewage sludge and city compost. Bull. Environ. Contam. Toxicol. 61, 347354. Ros, M., Hernandez, N.T., Garcia, C., 2003. Soil microbial activity after restoration of a semiarid soil by organic amendments. Soil Biol. Biochem. 35, 463469. SanPiN No 2.1.7.573-96, 1997. Gigienicheskie trebovaniya k ispolzovaniyu stichnich vod i ich osadkov dlya orosheniya i udobreniya M.: Informacionno-izdatelskii centr Minzdrava Rossii, Moskva (in Russian). Sanchez-Monedero, M.A., Mondini, C., Nobili, M., Leita, L., Roig, A., 2004. Land application of biosolids. Soil response to dierent stabilization degree of treated organic matter. Waste Manage. 24, 325332. Schinner, F. (Ed.), 1995. Methods in Soil Biology. Springer-Verlag, Berlin, Heidelberg. Selivanovskaya, S.Yu., Latypova, V.Z., Kiyamova, S.N., Alimova, F.K., 2001. Use of microbial parameters to assess treatment methods of municipal sewage sludge applied to grey forest soils of Tatarstan. Agri. Ecosyst. Environ. 86, 145153. Smith, S.R., 1994. Eect of soil pH on availability to crops of metals in sewage sludge treated soils. 1. Nickel, copper and zink uptake and toxicity to ryegrass. Environ. Pollut. 85, 321327. Wong, J.W.C., Lai, K.M., Su, D.S., Fang, M., 2001. Availability of heavy metals for Brassica chinensis grown in an acidic loamy soil amended with a domestic and industrial sewage sludge. Water Air Soil Pollut. 128, 339353.

eects on soil microorganisms were detected. In the soil amended with composted sewage sludge, Cmicr levels were always higher than those of the control soil. Some of the eects observed, particularly those on the pine seedlings indicate improvement of the soil quality. Even at very high application rates of 150 and 175 t ha1, no toxicity symptoms were exhibited in the pine seedling yields and plant behavior. The rates of composted sewage sludge from 75 to 100 t ha1 can be recommended to guarantee the best conditions for pine seedlings and the least eect on soil microbial population. In general, organic waste materials, e.g., composted sewage sludge appear stimulating for increasing the microbial biomass and activity of the grey forest soil of the forest nursery as well as for the germination and size of pine seedlings. Consequently, mineral fertilization can be reduced and partially substituted by organic waste materials. References
Anderson, T.H., Domsch, K.H., 1990. Application of eco-physiological quotients (qCO2 and qD) on microbial biomasses from soils of dierent cropping histories. Soil Biol. Biochem. 22, 251255. Anderson, T.H., Domsch, K.H., 1993. The metabolic quotient for CO2 (qCO2) as a specic activity parameter to assess the eects of environmental conditions. such as pH. on the microbial biomass of forest soils. Soil Biol. Biochem. 25, 393395. Chander, K., Brookes, P.C., 1991. Eects of heavy metals from past application of sewage sludge on microbial biomass and organic matter accumulation in a sandy loam and silty loam UK soil. Soil Biol. Biochem. 23, 927932. Chander, K., Brookes, P.C., 1993. Residual eects of zinc. copper and nikel in sewage sludge on microbial biomass in a sandy loam. Soil Biol. Biochem. 25, 12311239. Debosz, K., Petersen, S.O., Kure, L.K., Ambus, P., 2002. Evaluating eects of sewage sludge and household compost on soil physical. chemical and microbiological properties. Appl. Soil Ecol. 19, 237248. Gardiner, D.T., Miller, R.W., Badamchian, B., Azzari, A.S., Sisson, D.R., 1995. Eects of repeated sewage sludge application on plant accumulation of heavy metals. Agri. Ecosyst. Environ. 55, 16. Garcia-Gil, J.C., Plaza, C., Soler-Rovira, P., Polo, A., 2000. Long-term eects of municipal solid waste compost application on soil enzyme activities and microbial biomass. Soil Biol. Biochem. 32, 19071913. ISO 14240-2, 1997. International Standard. Soil quality determination of soil microbial biomass. Part 2: Fumigation-extraction method.