Soil Biology & Biochemistry

Utilization of Vermicomposts in Soil Restoration:

Effects on Soil Biological Properties
Manuel Tejada*
Isidoro Gmez
Dep. de Cristalografa, Mineraloga y
Qumica Agrcola
E.U.I.T.A. Univ. de Sevilla
Crta de Utrera km. 1
41013 Sevilla, Spain

Teresa Hernndez
Carlos Garca
Dep. de Conservacin de Suelos y Agua
y Manejo de Residuos Orgnicos
Centro de Edafologa y Biologa
Aplicada del Segura
CEBAS-CSIC
P.O. Box 4195
30080 Murcia, Spain

Increasingly, organic soil amendments are being examined for their potential use in soil restoration and for
preventing soil erosion. Two vermicomposts, differing in their chemical nature (obtained from cow dung, CD,
and green forage, GF), were applied annually for a period of 3 yr to a Xerollic Calciorthid soil located near
Seville (Guadalquivir Valley, Andalusia, Spain) to evaluate the efficiency of these organic amendments in soil
restoration. Their effects on the plant cover and biological properties (microbial biomass, soil respiration, and
enzymatic activities such as dehydrogenase, urease, -glucosidase, phosphatase, and arylsulfatase) of the soil were
determined. The organic wastes were applied at 3 and 6 Mg C ha1, respectively. After 3 yr of successive soil
amendment, the application of CD vermicompost to the soil had a greater effect on the soil biological properties
than GF vermicompost. The final soil microbial biomass C, dehydrogenase, urease, -glucosidase, phosphatase,
and arylsulfatase values were 28.3, 25.9, 12.6, 26, 12, and 14.2% higher in CD-amended soils than in the GFamended soils. This may have been due to a greater labile fraction of organic matter in the CD than the GF
vermicompost; however, the results obtained for the enzymatic activities stabilized in humic matrix mitigated that
the highest values occurred in the soils amended with GF vermicompost with respect to those amended with CD
vermicompost. This increased formation of enzymes immobilized in soils humic matrix may prolong any increase
in soil enzymatic activities and plant cover produced by the amendment.
Abbreviations: CD, cow dung; GF, green forage.

he soils of Mediterranean regions commonly have low organic matter content and are exposed to degradation, erosion, and desertification (Tejada and
Gonzlez, 2007). Recent years have seen an increase in the application of organic
wastes with a high organic matter content, usually composted, such as urban wastes
(Ros et al., 2003), plant materials derived from the municipal landscape (Walker,
2003), cotton gin compost (Tejada et al., 2006a), and beet vinasse composted with
a crushed cotton gin compost (Tejada et al., 2007), to semiarid soils for soil restoration purposes. Traditional methods of composting, however, can result in up to
55% of the organic matter and 30 to 50% of the N content being lost (Ketkar, 1993).
Vermicomposting, using earthworms, is recognized as an eco-biotechnological process that can transform complex organic substances into stabilized, humuslike products (Bentez et al., 2000). Earthworms accelerate the mineralization rate
and convert manures into casts with a higher nutritional value and degree of humification than the composts resulting from traditional methods of composting
(Albanell et al., 1988). The increased mineralization and conservation of nutrients
is due to the biocatalytic effect of earthworms in the decomposition and conservation mechanism (Suthar, 2007).
The extent to which vermicomposts influence soil properties and therefore
soil restoration or soil losses, however, depends on the amount, type, and structure (functional groups, molecular weight, humification, etc.) of the added organic
Soil Sci. Soc. Am. J. 74:525532
Published online 21 Jan. 2010
doi:10.2136/sssaj2009.0260
Received 8 July 2009.
*Corresponding author (mtmoral@us.es).
Soil Science Society of America, 677 S. Segoe Rd., Madison WI 53711 USA
All rights reserved. No part of this periodical may be reproduced or transmitted in any form or by
any means, electronic or mechanical, including photocopying, recording, or any information storage
and retrieval system, without permission in writing from the publisher. Permission for printing and for
reprinting the material contained herein has been obtained by the publisher.

SSSAJ: Volume 74: Number 2 MarchApril 2010

525

materials. Also, their effect on the maintenance of environmental

quality will depend on the substrates used during the vermicomposting process.
Tejada and Gonzlez (2009) found differences in soil biological properties and rice (Oryza sativa L.) quality and yield after the application of two different vermicomposts on an Aquic
Xerofluvent, observing that the application of animal-origin vermicompost resulted in a greater increase in soil biological properties and also in rice yield than did the application of vegetalorigin vermicompost.
The application of organic matter to degraded soils is a good
environmental practice. Therefore, the application of vermicompost to these soils has become a common environmental practice
for soil restoration, maintaining soil organic matter, reclaiming
degraded soils, and supplying plant nutrients. Also, the application of vermicomposts increases the vegetal cover and stimulates
soil microbial growth and activity (Arancon et al., 2006). The
stimulation of soil microbial activity, however, depends on the
organic matter chemical composition and, therefore, on the
vermicompost chemical composition. Biochemical and microbiological properties have traditionally been used to monitor
changes in soil quality and to quantify the effects of restoration
activities in degraded, semiarid soils (Prez de Mora et al., 2005;
Tejada et al., 2006b).
Microorganisms are largely responsible for the cycling of elements within a soil and are involved in the decomposition of organic matter at the ecosystem level. Parameters such as microbial
biomass C provide information on the size of these communities,
while others are related to their general activity, such as respiration or dehydrogenase activity, or are related specifically to the
cycling of elements in the soil, as is the case with urease, phosphatase,
-glucosidase, and arylsulphatase hydrolases (Garca et al., 2002).
The objective of this study was to evaluate the effects of adding two vermicomposts (one of animal and one of plant origin),
at different rates, on the soil biological activity and soil restoration in a semiarid Mediterranean ecosystem.

MATERIALS AND METHODS

Site Description and Properties of Vermicomposts
The study was conducted from October 2003 to October 2006
near Seville (Guadalquivir Valley, Andalusia, Spain), on a Xerollic
Table 1. Initial soil characteristics, with standard errors in
parentheses (data are the means of four samples).
Characteristic

Value

pH
7.6 (0.1)
Electrical conductivity, dS m1 0.22 (0.07)
Clay, g kg1
316 (14)
Silt, g kg1
256 (11)
Sand, g kg1
428 (13)
Texture
clay loam
Dominant clay type
illite, illitemontmorillonite (interstratified)
Bulk density, Mg m3
1.45 (0.04)
CaCO3, g kg1
341 (12)
Total N, g kg1
0.8 (0.03)
3.1 (0.08)
Total C, g kg1
526

Calciorthid soil (Soil Survey Staff, 1987) with an 8% slope. The general
properties of the soil (025 cm) are shown in Table 1.
The climate is semiarid, with an average annual precipitation of
400 mm for the three experimental years, concentrated in the spring
and autumn months. The mean annual temperature of the three experimental years was 17.3C and the mean potential evapotranspiration
was 700 mm yr1. Thus, the long-term water deficit, calculated by the
Thorntwaite method, is 436 mm. July and August are the driest months.
The area is a fragile environment, strongly marked by erosion.
Harsh physical conditions and inadequate soil use by humans have resulted in a dissected landscape where furrows, rills, and gullies scour
both the hillslopes and the weak deposits that fill the low-lying regions.
The substrates used to obtain the vermicomposts were cow dung
(CD) and green forage (GF), the latter basically consisting of grasses,
the green leaves of vegetables, herbs, and other plant materials. The general properties of both organic wastes, before initiating the vermicomposting process, are shown in Table 2. Organic C was determined by oxidation of C with potassium dichromate (Yeomans and Bremner, 1988),
while total N was determined by the Kjeldahl method (Ministerio de
Agricultura, Pesca y Alimentacin, 1986, p. 221285). After HNO3
HClO4 (1:1) acid digestion of both organic wastes, total P was determined by the method of Willians and Stewart, as described by Guitian
and Carballas (1976), and total K by atomic emission spectrometry.
Each substrate was vermicomposted on a bed consisting of a wooden frame (2 m high by 2 m wide by 3 m long) under aerobic conditions
(Bentez et al., 2002). Each bed was filled with 80 kg of substrate and inoculated with 800 adult epigeic earthworms (Eisenia fetida). The moisture level of both organic wastes was maintained at about 65 to 70%
during the vermicomposting period by periodically sprinkling them
with water. The vermicomposting process lasted 72 d for the CD and 85 d
for the GF, when the C/N ratio and the temperature had become constant.
The chemical composition of the vermicomposts at the end of the
vermicomposting process is shown in Table 3. To determine the humic
and fulvic acid C, the vermicomposts were extracted with a mixture of
0.1 mol L1 sodium pyrophosphate and 0.1 mol L1 NaOH at pH 13
(Kononova, 1966). The supernatant was acidified to pH 2 with HCl
and allowed to stand for 24 h at room temperature. To separate humic
acids from fulvic acids, the solution was centrifuged and the precipitate containing humic acids was dissolved with NaOH (Yeomans and
Bremner, 1988). After the removal of the humic acids, the acidic filtrate
containing the dissolved fulvic acid fraction was passed through a column of XAD-8 resin [(poly)methylmethacrylate]. The adsorbed fulvic
acid fraction was then recovered by elution with 0.1 mol L1 NaOH,
desalted using Amberlyst 15 cation-exchange resin (GFS Chemicals,
Table 2. Characteristics of the organic wastes before vermicomposting, with standard errors in parentheses (data are the
means of seven samples).
Cow dung
pH (1:10)
Total C, g kg1
Total N, g kg1
C/N ratio
Total P, g kg1
Total K, g kg1

6.7 (0.2)
388 (22)
6.8 (0.1)
57.1 (3.5)
6.4 (0.3)
7.2 (0.5)

Green forage
7.6 (0.1)
435 (17)
5.3 (0.2)
82.1 (2.8)
4.1 (0.2)
5.4 (0.3)

SSSAJ: Volume 74: Number 2 MarchApril 2010

Powell, OH), and finally freeze-dried. The C content of the humic acid and
fulvic acids was determined by the method of Yeomans and Bremner (1988).

Table 3. Characteristics of the vermicomposts, with standard

errors in parentheses (data are the means of seven samples).

Experimental Layout

pH (1:10)
6.4 (0.2)
364 (17)
Total C, g kg1
Humic acid C, g kg1
10.1 (0.3)
Fulvic acid C, g kg1
72.6 (3.5)
15 (2)
Total N, g kg1
C/N ratio
24.3 (1.2)
Total P, g kg1
10.4 (1.1)
11.3 (0.8)
Total K, g kg1

Cow dung

The experimental layout was a randomized, complete block design with five treatments and four replicates per treatment. The plot
size was 9 by 5 m, and the treatments were the following: (i) C, control
soil (no organic amendment); (ii) CD1, fertilized with 10.64 Mg ha1
of CD (3 Mg C ha1); (iii) CD2, fertilized with 21.28 Mg ha1 of
CD (6 Mg C ha1); (iv) GF1, fertilized with 7.71 Mg ha1 of GF
(3 Mg C ha1); and (v) GF2, fertilized with 15.42 Mg ha1 of GF
(6 Mg C ha1). The vermicomposts were applied to the soil surface on
16 Oct. 2003, 20 Oct. 2004, and 23 Oct. 2005 and incorporated to a
depth of 25 cm by chisel plowing and disking the day after application.
The chemical compositions of the vermicomposts remained unchanged
throughout the 3-yr experimental period. The original batch of vermicompost from CD and GF was kept refrigerated at 0C and thawed and
applied each year.

Soil Sampling and Analytical Determinations

Soil samples (025 cm) were collected from each plot with a gauge
auger (30-mm diameter) on 14 Oct. 2003, 19 Oct. 2004, 21 Oct. 2005,
and 20 Oct. 2006. Four subsamples were collected from each plot and
ground to pass through a 2-mm sieve before being stored in sealed polyethylene bags at 4C until analysis.
The soil pH was determined in distilled water with a glass electrode (1:2.5 soil/H2O ratio), as was the soil electrical conductivity (1:5
soil/H2O ratio). The soil texture was determined by Robinsons pipette
method (Soil Survey of England and Wales, 1982) and the dominant
clay types were determined by x-ray diffraction. The total CaCO3 was
measured by quantifying the CO2 produced by HCl addition to the soil
(Ministerio de Agricultura, Pesca y Alimentacin, 1986, p. 221285).
Soil organic C was determined by oxidizing the organic matter in the
soil samples with K2Cr2O7 in H2SO4 (96%) for 30 min and measuring the concentration of Cr3+ formed (Yeomans and Bremner, 1988).
The soil total N was determined by the Kjeldahl method (Ministerio
de Agricultura, Pesca y Alimentacin, 1986, p. 221285). The soil bulk
density was determined using the core method, weighing and drying
the soil at 105C for 48 h before determining the bulk density as the
ratio between the soil dry weight and the ring volume, according to the
official methods of the Spanish Ministry of Agriculture (Ministerio de
Agricultura, Pesca y Alimentacin, 1986, p. 221285) (Table 1).
The soil microbial biomass (SMB) was determined using the
CHCl3 fumigationextraction method (Vance et al., 1987). The C
content in the extract was measured using a C analyzer (TOC-5050A,
Shimadzu Corp., Kyoto, Japan), and SMB was calculated as SMB =
2.66(C extracted from fumigated samples C extracted from unfumigated samples) (Vance et al., 1987). The activity levels of five soil
enzymes were measured. Dehydrogenase activity was measured by reduction of 2-p-iodo-3-nitrophenyl 5-phenyl tetrazolium chloride to
iodonitrophenyl formazan (Garca et al., 1993). The urease activity was
determined by the buffered method of Kandeler and Gerber (1988), using urea as a substrate. Alkaline phosphatase activity was measured using
p-nitrophenyl phosphate as a substrate (Tabatabai and Bremner, 1969).

SSSAJ: Volume 74: Number 2 MarchApril 2010

Green forage
7.0 (0.1)
289 (14)
65.3 (2.1)
24.9 (2.8)
12 (1)
32.4 (1.9)
8.7 (0.6)
10.1 (0.9)

The -glucosidase activity was determined using p-nitrophenyl--dglucopyranoside as a substrate (Masciandaro et al., 1994). Arylsulfatase
activity was determined using p-nitrophenylsulfate as a substrate
(Tabatabai and Bremner, 1970).
Soil respiration was measured in the laboratory on 14 Oct. 2003,
19 Oct. 2004, 21 Oct. 2005, and 20 Oct. 2006 for each treatment. The
soil samples were incubated at 28C for a 90-d period in sealed flasks
containing a vial of NaOH solution. The NaOH vials containing the
CO2C evolved from the soil samples were replaced for analysis after
3, 7, 15, 30, 60, and 90 d of incubation. The CO2C collected in the
NaOH was determined by the addition of an excess of 1.5 mol L1 BaCl2,
followed by titration with standardized HCl using a phenolphthalein
indicator (Zibilske, 1994).
The enzymatic activities of urease, phosphatase, -glucosidase, and
arylsulfatase were measured in the 0.1 mol L1 sodium pyrophosphate
extract to estimate the enzymatic activity immobilized in the humic
matrix of the soil and the effect of organic amendment on this fraction
of soil enzymatic activity. The extraction was performed in the cold for
5 h, following sonication of a 1:10 soil/pyrophosphate mixture. The
extract was separated by centrifugation at 10,000 rpm for 30 min and
filtered through Millipore 0.22-m membrane filters (microbiological
filtration). Finally, an extract of an aliquot was dialyzed against distilled
water in cut molecular weight membranes of 14,000 units (Visking dialysis tube, Serva, Germany). The extraction was performed in triplicate.
Urease, phosphatase, -glucosidase, and arylsulfatase activities were determined in the extracts using the methods described above.
Plant cover, or the percentage of the soil covered by the octagonal
projection of the aerial part of each plant, was determined by the linear
intercept method (Canfield, 1941).

Statistical Analysis
Analysis of variance was performed using the Statgraphics 5.0 software package (Statistical Graphics Corporation, 1991). The means were
separated by Tukeys test, considering a significance level of P < 0.05
throughout the study.
The data were subjected to a correlation matrix to obtain the parameters that most influenced plant cover. Lastly, linear regression analysis
was performed with the parameters identified in the correlation matrix.

RESULTS
Soil Properties
The soil respiration values (CO2C evolved during the 90-d
of incubation) for each experimental year are shown in Table 4.

527

Table 4. Cumulative CO2C during 90-d incubation of soils amended with vermicom- The CO C emissions were higher
2
posts, at the end of the first, second, and third experimental year.

in CD2, followed by the GF2, CD1,

GF1, and C treatments, indicating that
3d
7d
15 d
30 d
60 d
90 d
the effect of the vermicomposts on the
mg kg1 soil
soil depended on the rate and type of
2004
organic amendment. The CO2C emisC
138a 17 311ab 22 417b 35
579b 42
725bc 55
793c 41
sions
were higher at the end of the exCD1
272ab 24 569b 32 883c 44
1107c 51 1311cd 44 1706d 27
perimental
period than during the first
CD2
355ab 26 669bc 41 1022c 52 1321cd 38
1553d 39 1963d 46
year, possibly due to the residual effect of
GF1
247a 22 504b 19 803c 28
1032c 79 1209cd 48 1582d 55
GF2
301ab 17 608b 23 969c 47 1211cd 82
1432d 66 1844d 91
the organic matter on the soil. At the end
2005
of the incubation period, the statistical
C
130a 14 301ab 34 402b 44
560b 42
710bc 57 779bc 32
analyses pinpointed significant differences
CD1
304ab 22 601b 47 943c 51
1207c 60 1422cd 39 1813d 51
between all organic treatments and the
CD2
411b 34 722bc 49 1133c 35 1422cd 29
1664d 38 2073d 49
control soil.
GF1
275ab 28 539b 55 849c 24
1122c 29 1307cd 24 1673d 74
Soil microbial biomass increased
GF2
341ab 20 667bc 41 1067c 32 1322cd 53
1528d 40 1962d 93
during the experimental period in the
2006
vermicompost-amended soils, the greatC
122a 13 290ab 29 390b 19
549b 40
695bc 62 762bc 29
est increase being recorded in the soil
CD1
339ab 27 629b 38 1006c 27 1350cd 37
1556d 55 1924d 46
amended with the higher dose of CD
CD2
462b 38 799c 45 1244c 88
1511d 44
1749d 69 2193d 93
vermicompost (Fig. 1). At the end of
GF1
310ab 40 569b 34 893c 79
1249c 59 1399cd 34 1755d 88
the experimental period and for the
GF2
395b 33 721bc 53 1149c 68
1438d 40
1627d 67 2011d 105
C, control soil; CD1, fertilized with 10.64 Mg cow dung ha1; CD2, fertilized with
higher rates of organic matter applied
21.28 Mg cow dung ha1; GF1, fertilized with 7.71 Mg green forage ha1; GF2, fertilized with
to the soil, the soil microbial biomass in
1
15.42 Mg green forage ha .
soils amended with CD vermicompost
Means SE.
was (significantly) 21.8% higher than
in soils amended with GF vermicompost. The control soil showed the lowest soil
microbial biomass values of all the treatments
assayed throughout the experimental period.
The highest enzyme activities at the
end of the experiment were observed in soils
amended with CD vermicompost, followed
by the soils amended with GF vermicompost
and the control soil (Fig. 1 and 2). In this respect, dehydrogenase, urease, -glucosidase,
phosphatase, and arylsulfatase activities increased 25.9, 12.6, 26, 12, and 14.2%, respectively, in the soils amended with CD vermicompost, with respect to the soils amended
with GF vermicompost. The statistical analysis pointed out significant differences in the
soil enzymatic activities between the two organic treatments for the dehydrogenase, urease, and -glucosidase activities.
The enzymatic activities stabilized in the
humic matrix evolved differently from the soil
enzymes not adsorbed to humus (Fig. 3). At
the end of the experimental period, the urease immobilized had increased by 69.6, 63.8,
61.3, and 54.5% over the control soil in GF2,
CD2, GF1, and CD1, respectively. The statisFig. 1. Microbial biomass C, dehydrogenase activity, and urease activity in soils to which organic
1
wastes were applied. Treatments were: C, control soil; CD1, fertilized with 10.64 Mg cow dung ha ; tical analysis again showed significant differCD2, fertilized with 21.28 Mg cow dung ha1; GF1, fertilized with 7.71 Mg green forage ha1; ences between GF2 and CD2 treatments at
and GF2, fertilized with 15.42 Mg green forage ha1. Columns (mean SE) followed by the same the end of the experiment. Also, at the end of
Treatment

Cumulative CO2C

letter(s) are not significantly different (P < 0.05); INTF is 2-p-iodo-3-nitrophenyl.

528

SSSAJ: Volume 74: Number 2 MarchApril 2010

the experimental period, the -glucosidase immobilized in the humic matrix had increased
by 25.1, 39.7, and 62.3% in the GF2 treatment
compared with the CD2, GF1, and CD1 treatments, respectively. The phosphatase immobilized showed very similar behavior, being 13.3,
22.2, and 44.2% higher in GF2 than in CD2,
GF1, and CD1, respectively. Lastly, also at the
end of the experiment, the arylsulfatase stabilized was 17.8, 35.8, 48.7, and 97.4% greater in
GF2 than in CD2, GF1, CD1, and the control
soil, respectively. The ANOVA analysis indicated no significant differences between the two
organic treatments at the end of the experimental period.

Plant Cover
One year after the first organic amendment, the treated plots were covered with spontaneous vegetation, with the most abundant
being Borago officinalis L., Chrysanthemum
coronarium L., Diplotaxis muralis (L.) DC.,
Paronychia argentea Lam., and Silene colorata Poiret. After 3 yr, the percentage of plant
cover decreased in the following order: CD2
plot (88% plant cover) > GF2 plot (71% plant
cover) > CD1 plot (60% plant cover) > GF1
plot (45% plant cover) > control soil (9% plant
cover) (Fig. 4). The ANOVA indicated signifi-

Fig. 2. The -glucosidase, phosphatase, and arylsulfatase activities in soils to which

organic wastes were applied. Treatments were: C, control soil; CD1, fertilized with
10.64 Mg cow dung ha1; CD2, fertilized with 21.28 Mg cow dung ha1; GF1, fertilized with
7.71 Mg green forage ha1; and GF2, fertilized with 15.42 Mg green forage ha1. Columns
(mean SE) followed by the same letter(s) are not significantly different (P < 0.05); PNP is
p-nitrophenol and PNF is p-nitrophenyl.

Fig. 3. The activity of urease, -glucosidase, phosphatase, and arylsulfatase immobilized in the humic matrix of soils to which vermicomposts
were applied. Treatments were: C, control soil; CD1, fertilized with 10.64 Mg cow dung ha1; CD2, fertilized with 21.28 Mg cow dung ha1;
GF1, fertilized with 7.71 Mg green forage ha1; and GF2, fertilized with 15.42 Mg green forage ha1. Columns (mean SE) followed by the same
letter(s) are not significantly different (P < 0.05); PNP is p-nitrophenol and PNF is p-nitrophenyl.
SSSAJ: Volume 74: Number 2 MarchApril 2010

529

The higher microbial biomass

and respiration measured in the soils
amended with the vermicompost
with a higher fulvic acid content
may have been due to the greater
labile fraction of organic matter in
the this product. The labile fraction of organic matter is the most
degradable and, therefore, the most
susceptible to mineralization (Cook
and Allan, 1992), acting as an immediate energy source for microorFig. 4. Plant cover on soils to which organic wastes were applied. Columns (mean SE) followed by the same ganisms. Curves representing cumuletter(s) are not significantly different (P < 0.05).
lative CO2C evolution with time
cant differences between the CD2 and GF2 treatments at the
show that the slope at the outset was
end of the experiment.
higher in the soil amended with the CD vermicompost, which
To predict the parameters influencing plant cover most decisuggests that the C substrates were mineralized more rapidly in
sively, the variables were subjected to a correlation matrix (Table
this treatment, while the greater microbial biomass derived from
5). Since plant cover was related to the soil biological properties
this treatment would have been able to degrade a greater quantity
studied, a linear regression analysis was performed, considerof substrate (Table 4).
ing plant cover as the dependent variable. The resulting regresSoil microorganisms degrade organic matter through the
sion equation was: Plant cover = 7.9712 + 0.081 biomass C +
production of a variety of extracellular enzymes, which explains
0.2506 dehydrogenase + 0.6725 urease 0.4491 -glucosidase
the increase in enzymatic activities observed after the application
0.3905 phosphatase + 1.2556 arylsulfatase + 13.275 humus
of vermicomposts to the soil. These results are in agreement with
urease + 8.21565 humus-glucosidase 2.7086 humusphosthose of Arancon et al. (2006) and Ferreras et al. (2006), who
2
phatase + 1.1353 humusarylsulfatase; R = 97.34%. The high
also found an increase in enzymatic activities after the addition
correlation coefficient indicates a strong correlation between
of different vermicomposts to soil. Soil enzymes act as biological
plant cover and these variables.
catalysts of specific reactions that depend on a variety of factors,
such as the presence or absence of inhibitors, amendment type,
DISCUSSION
and crop type, and can be considered as early indicators of bioOur results indicate that amendment with the vermicomlogical changes (Bandick and Dick, 1999). The incorporation of
post that had a higher fulvic acid content in its chemical comorganic amendments into the soil influences the soil enzymatic
position was more likely to favor soil biological properties and
activities because the added material may contain intra- and explant cover. Obviously, the supply of readily metabolizable C in
tracellular enzymes and may also stimulate microbial activity in
the organic waste products will be the most influential factor in
the soil (Goyal et al., 1999; Pascual et al., 1998).
the measured increase in biomass C. According to Blagodatsky
Any increase in soil microbial biomass C, soil respiration, or
et al. (2000), Tejada et al. (2006a), and Tejada et al. (2007), the
soil enzymatic activities will vary considerably, however, dependsoil microbial biomass responds rapidly to additions of readily
ing on the type of vermicompost applied to the soil. Because the
available C.
C substrates in CD are mineralized in the soil more rapidly than those

Table 5. Correlation matrix of the soil biological properties studied during the experimental period. All correlations are significant
at the 0.001 level.
Property
MBC
DHA
URA
GLUA
PHOA
ARA
HURA
HGLUA
HPHOA
HARA
PC
MBC
1.000
DHA
0.962
1.000
URA
0.967
0.993
1.000
GLUA
0.965
0.988
0.992
1.000
PHOA
0.977
0.986
0.994
0.988
1.000
ARA
0.922
0.956
0.961
0.951
0.963
1.000
HURA
0.916
0.938
0.944
0.947
0.953
0.947
1.000
HGLUA
0.889
0.937
0.941
0.934
0.931
0.916
0.939
1.000
HPHOA
0.871
0.905
0.911
0.909
0.903
0.884
0.914
0.988
1.000
HARA
0.908
0.907
0.913
0.916
0.903
0.875
0.918
0.969
0.975
1.000
PC
0.978
0.980
0.982
0.973
0.985
0.967
0.949
0.910
0.874
0.896
1.000
MBC, microbial biomass C; DHA, dehydrogenase activity; URA, urease activity; GLUA, -glucosidase activity; PHOA, phosphatase activity;
ARA, arylsulfatase activity; HURA, humusurease complex; HGLUA, humus-glucosidase complex; HPHOA, humusphosphatase complex;
HARA, humusarylsulfatase complex; PC, plant cover.
530

SSSAJ: Volume 74: Number 2 MarchApril 2010

in GF, the enzymatic activities in soils amended with CD vermicompost were higher than in soils amended with GF vermicompost.
The stabilization of enzyme activities in the humic matrix
also may contribute to the higher hydrolase enzyme activity
measured in soils amended with vermicomposts having a higher
fulvic acid content. According to Nannipieri et al. (1996), the
formation of enzymes in the humic matrix occurs mainly for
the humic acid fraction. Humic and fulvic acids differ in their
structures. Fulvic acids are macromolecules with a lower polymerization index than humic acids and are more highly charged,
more polar, and have a lower molecular weight. Hayes (1991)
suggested that humic substances with higher molecular weight
fractions contain more strong acid groups than lower molecular
weight materials. Humic-like acids have greater aromaticity than
fulvic-like acids, which is also in keeping with the concept of
greater numbers of aromatic carboxylic acids in the humic acids.
For this reason, the immobilization of enzymes in the humic matrix may be more common for humic-like than for fulvic-like acids.
Burns (1982) suggested that enzymes immobilized on humus molecules play an important role in soil microbial ecology.
Immobilized enzymes may act as stable catalysts for the detection
of potential substrates, making the continuous synthesis and secretion of extracellular enzymes by microorganisms unnecessary.
Microorganisms can synthesize and release the extracellular enzyme only if the substrate is present and the environmental conditions are suitable for catalysis. Therefore, these results suggest
that vermicomposts rich in humic-like acids are preferable if the
aim is to prolong any increase in soil enzymatic activities. Free
enzymes normally show short-lived activity because they are rapidly denatured, degraded, or inhibited irreversibly. Extracellular
enzymes are usually associated with, and protected by, soil colloids, such as clays or humic substances (Nannipieri et al., 1996)
and act as a stable nucleus of soil activity. These humus enzymes
exhibit great resistance against thermal denaturation, dehydration, and proteolysis (Nannipieri et al., 1996).
Because soil enzymatic activities are responsible for important cycles such as those of C, N, P, and S, the plant cover increased significantly with the higher dose of CD vermicompost.
The values were similar to those reported by Tejada et al. (2006a)
after the addition of crushed cotton (Gossypium hirsutum L.)
gin compost and poultry manure under similar pedoclimatic
conditions. The plant cover in the third year was higher than in
the previous 2 yr due to the residual effect of the organic matter of each vermicompost waste after their application in the first
and second experimental seasons. It is necessary to emphasize,
however, that an increase in the formation of humusenzyme
complexes in the soils amended with vermicomposts with a higher humic acid concentration may act by prolonging any increase in the
soil enzymatic activities and plant cover for a longer time.
These results are very important, principally in the arid
zones, where the risk of desertification and loss of soil is a real
problem (Tejada et al., 2006a, 2007).

SSSAJ: Volume 74: Number 2 MarchApril 2010

CONCLUSIONS
The application of vermicomposts at the doses studied,
under dryland conditions, caused an improvement in the soil
biological properties and plant cover. Consequently, the addition of these organic wastes may be considered a good strategy
for recovering semiarid areas. The soil biological properties and
plant cover were better in soils amended with the CD vermicompost, rich in fulvic-like acids, possibly due to the greater labile
fraction of organic matter than in the GF vermicompost, making it more degradable and susceptible to rapid mineralization.
Nevertheless, the increased enzymatic activities immobilized in
the humic matrix in soils amended with GF vermicompost, with
respect to soils amended with CD vermicompost, suggests that
vermicomposts rich in humic-like acids are more suitable for prolonging any increase in plant cover. Because plant cover is related
directly to the soil biological properties, a study of the soil enzymatic activities might be a good indicator of the possible evolution of the plant cover.

REFERENCES
Albanell, E., J. Plaixats, and T. Cabreo. 1988. Chemical changes during
vermicomposting (Eisenia fetida) of sheep manure mixed with cotton
industrial wastes. Biol. Fertil. Soils 6:266269.
Arancon, N.Q., C.A. Edwars, and P. Bierman. 2006. Influences of vermicomposts
on field strawberries: 2. Effects on soil microbiological and chemical
properties. Bioresour. Technol. 97:831840.
Bandick, A.K., and R.P. Dick. 1999. Field management effects on soil enzymes
activities. Soil Biol. Biochem. 31:14711479.
Bentez, E., R. Nogales, G. Masciandaro, and B. Ceccanti. 2000. Isolation by
isoelectric focusing of humicurease complexes from earthworm (Eisenia
fetida)-processed sewage sludges. Biol. Fertil. Soils 31:489493.
Bentez, E., H. Sainz, R. Melgar, and R. Nogales. 2002. Vermicomposting of a
lignocellulosic waste from olive oil industry: A pilot scale study. Waste
Manage. Res. 20:134142.
Blagodatsky, S., O. Heinemeyer, and J. Richter. 2000. Estimating the active and
total soil microbial biomass by kinetic respiration analysis. Biol. Fertil. Soils
32:7381.
Burns, R.G. 1982. Enzyme activity in soil: Location and a possible role in
microbial ecology. Soil Biol. Biochem. 14:423427.
Canfield, R.H. 1941. Application of the line intercept methods in sampling
range vegetation. J. For. 39:384388.
Cook, B.D., and D.L. Allan. 1992. Dissolved organic matter in old field soils:
Total amounts as a measure of available resources for soil mineralization.
Soil Biol. Biochem. 24:585594.
Ferreras, L., E. Gmez, S. Toresani, L. Firpo, and R. Rotondo. 2006. Effect of
organic amendments on some physical, chemical and biological properties
in a horticultural soil. Bioresour. Technol. 97:635640.
Garca, C., T. Hernndez, F. Costa, B. Ceccanti, and G. Masciandaro. 1993.
The dehydrogenase activity of soils and ecological marker in processes
of perturbed system regeneration. p. 89100. In Int. Symp. Environ.
Biogeochem., 11th, Salamanca, Spain. 27 Sept.1 Oct. 1993. Centro de
Informacion y Documentacion Cientifica, Madrid.
Garca, C., T. Hernndez, A. Roldn, and A. Martn. 2002. Effect of plant cover
decline on chemical and microbiological parameters under Mediterranean
climate. Soil Biol. Biochem. 34:635642.
Goyal, S., K. Chander, M.C. Mundra, and K.K. Kapoor. 1999. Influence of inorganic
fertilizers and organic amendments on soil organic matter and soil microbial
properties under tropical conditions. Biol. Fertil. Soils 29:196200.
Guitian, F., and T. Carballas. 1976. Tcnicas de anlisis de suelos. Picro Sacro,
Santiago de Compostela, Spain.
Hayes, H.B.H. 1991. Concepts of the origins, composition, and structures of
humic substances. p. 322. In W.S. Wilson (ed.) Advances in soil organic
matter research: The impact on agriculture and the environment. R. Soc.
Chem., Cambridge, UK.

531

Kandeler, E., and H. Gerber. 1988. Short-term assay of soil urease activity using
colorimetric determination of ammonium. Biol. Fertil. Soils 6:6872.
Ketkar, C.M. 1993. Use of biogas slurry in agriculture. p. 2426. In Biogas slurry
utilization. Consortium on Rural Technology, New Delhi.
Kononova, M.M. 1966. Soil organic matter. 2nd ed. Pergamon Press, Oxford, UK.
Masciandaro, G., B. Ceccanti, and C. Garca. 1994. Anaerobic digestion of straw and
piggery waste waters: II. Optimization of the process. Agrochimica 38:195203.
Ministerio de Agricultura, Pesca y Alimentacin. 1986. Mtodos oficiales de
anlisis. Vol. 1. Ministerio de Agricultura, Pesca y Alimentacin, Madrid.
Nannipieri, P., P. Sequi, and P. Fusi. 1996. Humus and enzyme activity. p.
293328. In A. Piccolo (ed.) Humic substances in terrestrial ecosystems.
Elsevier, Amsterdam.
Pascual, J.A., T. Hernndez, C. Garca, and M. Ayuso. 1998. Enzymatic activities
in an arid soil amended with urban organic wastes: Laboratory experiment.
Bioresour. Technol. 64:131138.
Prez de Mora, A., J.J. Ortega-Calvo, F. Cabrera, and E. Madejn. 2005. Changes
in enzyme activities and microbial biomass alter in situ remediation of a
heavy-metal contaminated soil. Appl. Soil Ecol. 28:125137.
Ros, M., M.T. Hernndez, and C. Garca. 2003. Soil microbial activity after restoration
of a semiarid soil by organic amendments. Soil Biol. Biochem. 35:463469.
Soil Survey of England and Wales. 1982. Soil survey laboratory methods. Tech.
Monogr. 6. Soil Survey of England and Wales, Harpenden, UK.
Soil Survey Staff. 1987. Keys to Soil Taxonomy. Soil Manage. Support Serv. Tech.
Monogr. 6. 3rd ed. Cornell Univ., Ithaca, NY.
Statistical Graphics Corporation. 1991. Statgraphics 5.0 statistical graphics
system. Statistical Graphics Corp., Warrenton, VA.
Suthar, S. 2007. Vermicomposting potential of Perionyx sansibaricus (Perrier) in
different waste materials. Bioresour. Technol. 98:12311237.

532

Tabatabai, M.A., and J.M. Bremner. 1969. Use of p-nitrophenol phosphate for
assay of soil phosphatase activity. Soil Biol. Biochem. 1:301307.
Tabatabai, M.A., and J.M. Bremner. 1970. Arylsulfatase activity of soils. Soil Sci.
Soc. Am. Proc. 34:225229.
Tejada, M., C. Garca, J.L. Gonzlez, and M.T. Hernndez. 2006a. Organic
amendment based on fresh and composted beet vinasse: Influence on soil
properties and wheat yield. Soil Sci. Soc. Am. J. 70:900908.
Tejada, M., and J.L. Gonzlez. 2007. Influence of organic amendments on soil
structure and soil loss under simulated rain. Soil Tillage Res. 93:197205.
Tejada, M., and J.L. Gonzlez. 2009. Application of two vermicomposts on a rice
crop: Effects on soil biological properties and rice quality and yield. Agron.
J. 101:336344.
Tejada, M., M.T. Hernndez, and C. Garca. 2006b. Application of two organic
amendments on soil restoration: Effects on the soil biological properties. J.
Environ. Qual. 35:10101017.
Tejada, M., J.L. Moreno, M.T. Hernndez, and C. Garca. 2007. Application of
two beet vinasse forms in soil restoration: Effects on soil properties in an arid
environment in southern Spain. Agric. Ecosyst. Environ. 119:289298.
Vance, E.D., P.C. Brookes, and D.S. Jenkinson. 1987. An extraction method for
measuring soil microbial biomass C. Soil Biol. Biochem. 19:703707.
Walker, R.F. 2003. Comparison of organic and chemical soil amendments used in
the reforestation of a harsh Sierra Nevada site. Restor. Ecol. 11:446474.
Yeomans, J.C., and J.M. Bremner. 1988. A rapid and precise method for routine
determination of organic carbon in soil. Commun. Soil Sci. Plant Anal.
19:14671476.
Zibilske, L.M. 1994. Carbon mineralization. p. 835863. In R.W. Weaver et al.
(ed.) Methods of soil analysis. Part 2. Microbiological and biochemical
properties. SSSA Book Ser. 5. SSSA, Madison, WI.

SSSAJ: Volume 74: Number 2 MarchApril 2010