Science of the Total Environment 563564 (2016) 160168

Contents lists available at ScienceDirect

Science of the Total Environment

journal homepage: www.elsevier.com/locate/scitotenv

Loss of soil (macro)fauna due to the expansion of Brazilian

sugarcane acreage
Andr L.C. Franco a,e,, Marie L.C. Bartz b, Maurcio R. Cherubin a, Dilmar Baretta c, Carlos E.P. Cerri d,
Brigitte J. Feigl a, Diana H. Wall e, Christian A. Davies f, Carlos C. Cerri a
a

Center for Nuclear Energy in Agriculture, University of So Paulo, Av. Centenrio 303, 13416000, Piracicaba, SP, Brazil
Universidade Positivo, Rua Prof. Pedro de Souza 5300, 81280330, Curitiba, PR, Brazil
Santa Catarina State University, Rua Beloni Zanin 680E, 89815630, Chapec, SC, Brazil
d
Department of Soil Science, Luiz de Queiroz College of Agriculture, University of So Paulo, Av. Pdua Dias 11, 13418900, Piracicaba, SP, Brazil
e
School of Global Environmental Sustainability & Department of Biology, Colorado State University, 80523, Fort Collins, CO, USA
f
Shell Technology Centre Houston, 3333 Highway 6 South, Houston, TX 77082, USA
b
c

H I G H L I G H T S

G R A P H I C A L

A B S T R A C T

We examined the soil macrofaunal

community responses to sugarcane expansion from pasturelands.
Macrofaunal abundance decreases 89%
and diversity of macrofauna groups deceases 39% when sugarcane replaces
pasture.
Termites and earthworms largely dominate the macrofaunal community in PA
soils.
Ants and coleopteran insects become
the most abundant groups when land
use is changed to SC.
This study draws attention for a signicant biodiversity loss belowground due
to bioenergy crop expansion in the tropics.

a r t i c l e

i n f o

Article history:
Received 17 November 2015
Received in revised form 5 April 2016
Accepted 17 April 2016
Available online 30 April 2016
Editor: D. Barcelo
Keywords:
Bioenergy crops
Land use change
Soil biodiversity

a b s t r a c t
Land use changes (LUC) from pasture to sugarcane (Saccharum spp.) crop are expected to add 6.4 Mha of new
sugarcane land by 2021 in the Brazilian Cerrado and Atlantic Forest biomes. We assessed the effects of these
LUC on the abundance and community structure of animals that inhabit soils belowground through a eld survey
using chronosequences of land uses comprising native vegetation, pasture, and sugarcane along a 1000-km-long
transect across these two major tropical biomes in Brazil. Macrofauna community composition differed among
land uses. While most groups were associated with samples taken in native vegetation, high abundance of
termites and earthworms appeared associated with pasture soils. Linear mixed effects analysis showed that
LUC affected total abundance (X2(1) = 6.79, p = 0.03) and taxa richness (X2(1) = 6.08, p = 0.04) of soil macrofauna. Abundance increased from 411 70 individuals m2 in native vegetation to 1111 202 individuals m2
in pasture, but decreased sharply to 106 24 individuals m2 in sugarcane soils. Diversity decreased 24% from
native vegetation to pasture, and 39% from pasture to sugarcane. Thus, a reduction of ~90% in soil macrofauna
abundance, besides a loss of ~ 40% in the diversity of macrofauna groups, can be expected when sugarcane

Corresponding author at: Department of Biology, Colorado State University, 80523, Fort Collins, CO, USA.
E-mail address: andre.franco@colostate.edu (A.L.C. Franco).

http://dx.doi.org/10.1016/j.scitotenv.2016.04.116
0048-9697/ 2016 Elsevier B.V. All rights reserved.

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

161

crops replace pasture in Brazilian tropical soils. In general, higher abundances of major macrofauna groups (ants,
coleopterans, earthworms, and termites) were associated with higher acidity and low contents of macronutrients
and organic matter in soil. This study draws attention for a signicant biodiversity loss belowground due to tropical LUC in sugarcane expansion areas. Given that many groups of soil macrofauna are recognized as key mediators of ecosystem processes such as soil aggregation, nutrients cycling and soil carbon storage, our results warrant
further efforts to understand the impacts of altering belowground biodiversity and composition on soil functioning and agriculture performance across LUC in the tropics.
2016 Elsevier B.V. All rights reserved.

1. Introduction
One of the central sustainability issues related to the rapid growth of
bioenergy industry refers to the implications of large-scale biofuel feedstock cultivation on biodiversity (Dauber and Bolte, 2014; Wiens et al.,
2011). Land use changes (LUC) associated with bioenergy feedstock
production appear as a key driver of above and belowground biodiversity changes due to the demand for biofuels, and the negative impacts
are primarily reported from tropical regions (Desiree et al., 2014;
McCormack et al., 2013). In Brazil, sugarcane (Saccharum spp.) is the
single plant used for ethanol fuel production and the additional amount
of land under sugarcane necessary to meet the domestic demand for
ethanol for 2021 is projected to be 6.4 Mha (Goldemberg et al., 2014),
which is larger than the total area of countries such as Netherlands
or Switzerland. Typical LUC in Brazilian sugarcane expansion areas
are the conversion of cultivated pastures into sugarcane elds
(Goldemberg et al., 2014). Roughly 70% of the new sugarcane elds
established between 2000 and 2010 in Brazil were previously cultivated
pastures used for cattle ranching (Adami et al., 2012). Those pastures
mostly replaced native vegetation areas between 1970 and 1980 by a
conversion that involves clearing native vegetation through traditional
slash and burn practices, and planting pasture grass in the same year.
Conversion of native vegetation directly into sugarcane have historically
accounted for less than 1% of the sugarcane expansion in Brazil (Adami
et al., 2012).
The existing literature does not show a clear position about the
effects of direct LUC for sugarcane expansion on biodiversity
(Goldemberg et al., 2008; Verdade et al., 2012). Particularly the responses of soil biodiversity to these LUC remain unexplored. Soil animals are estimated to represent as much as 23% of the total described
global diversity (Decaens et al., 2006). The larger soil invertebrates are
the soil macrofauna, with a body length longer than 2 mm (Swift
et al., 1979). Soil macrofauna are key mediators of ecosystem functions
for a variety of ecosystem engineering processes (Lavelle et al., 2006).
Soil macroinvertebrates actively inuence pedological processes
through structural and mineralogical transformations (de Oliveira
et al., 2014), and improve soil hydraulic properties such as aeration
and drainage (Leonard and Rajot, 2001). These animals also have an acknowledged role in building and stabilizing soil aggregates with direct
implications on ood and erosion control (Brussaard et al., 2007; Six
et al., 2004), soil carbon storage and net soil greenhouse-gas emissions
(Lubbers et al., 2013; Majeed et al., 2014). Moreover, these animals signicantly inuence nutrient cycling processes (de Vries et al., 2013;
Lubbers et al., 2013; Wagg et al., 2014).
We followed the progress of modications in soil macrofauna abundance and its community structure across LUC through a eld survey
using chronosequences of land use over two major tropical biomes in
Brazil. We predicted that sugarcane establishment over pasturelands
would reduce the soil macrofauna abundance as well as the diversity
of macrofauna groups. Rationale for this hypothesis is that the intensied land use in sugarcane crops compared to pastures would substantially reduce the quality and quantity of inputs available as well as the
soil micro-habitats, nally leading to loss of soil animals and simplication of the soil community, as it has been observed in other
agroecosystems (Callaham et al., 2006; Castellanos-Navarrete et al.,
2012; Hernandez-Ruiz and Castano-Meneses, 2006). Every ve years a

cultivation cycle is carried in sugarcane elds with plowing and fertilization for planting of new stem cuttings, reducing soil carbon stocks over
time (Franco et al., 2015; Mello et al., 2014) and potentially altering soil
structure and bulk density. Sugarcane farming is also characterized by
signicant inputs of persistent herbicides and controlled release pesticides (Bell et al., 2007). On the other hand, LUC from pasture to sugarcane promote signicant improvements on soil chemical quality by
increasing nutrient levels and reducing soil acidity due to inputs of fertilizers and lime (Cherubin et al., 2015), what gives rise to the hypothesis that detrimental LUC effects on soil macrofauna community would
be inversely related to soil chemical quality.
Our goal was to evaluate the effects of LUC to the expansion of
Brazilian sugarcane acreage on the soil macrofaunal community. Specifically, we wanted: (1) to compare soil macrofauna abundance, taxonomic richness, diversity and equability among different land uses;
(2) to investigate the vertical distribution of soil macrofauna abundance
and richness within the soil pedon as a function of LUC; (3) to explore
the relationships between macrofauna biodiversity and soil chemical attributes (soil acidity, and contents of soil organic carbon (SOC) and
macronutrients).
2. Materials and methods
2.1. Description of the study sites
The study was carried out in the main sugarcane production region
in the world, Central-Southern Brazil. Three study sites were identied
representing the Northern, Central, and Southern parts of the Brazilian
sugarcane growing region, including the areas where sugarcane expansion is occurring from pastures: Northern, Lat_17S, located in the city of
Jata, Southwestern region of Gois state (175616S, 513831W);
Central, Lat_21S, located in the city of Valparaso, West region of So
Paulo state (211448S, 504704W); and Southern, Lat_23S, located
in the city of Ipaussu, South region of So Paulo state (230508S,
493752W) (Fig. 1).
In each study site we identied a chronosequence of land use for: native vegetation, pasture, and sugarcane crop, and to minimize the effects
of climatic, topographic and edaphic variations, the three land uses were
always located in adjacent areas within sites. Climate and soil characteristics can be found in Table 1. A complete soil chemical characterization
can be found in Cherubin et al. (2015), and detailed information on the
past history of land use and soil management in the study sites is provided in Table A.1. Briey, native vegetation at Lat_17S comprises the
Cerrado forest formation, while at Lat_21S and Lat_23S it comprises a
transition between the Atlantic forest and Cerrado vegetation. LUC
from native vegetation to pasture happened in 1980 at both Lat_17S
and Lat_21S, and in 1979 at Lat_23S. Pasture areas differed from each
other in the stocking rate: 1.5 animal unit (AU, or a 454 kg cow with
or without an unweaned calf) ha1 at Lat_17S; around 2 AU ha1 at
Lat_21S; and around 1 AU ha1 at Lat_23S along the year. Sugarcane
was established over part of the pasture area in 2009 at Lat_17S, in
2010 at Lat_21S, and in 1990 at Lat_23S. The nutrient inputs in sugarcane differed among the sites, with annual inputs of mineral P at
Lat_17S and Lat_21S, and high amounts of organic fertilizers at
Lat_23S. The temporal differences in LUC time added to the contrastive
soil types among sites enlarged the strength of the data set taken.

162

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

Fig. 1. Location of the study sites in Brazil.

2.2. Sampling
The eld sampling for soil macrofauna was carried out in the rainy
season, in January 2013. High soil fauna richness and reduced variation
of abundance are expected during the rainy season, which is therefore
more indicated for the assessment of soil fauna (Neto et al., 2012). All
samples from a single site were taken on the same day. At each site macrofauna samples were extracted from a 1-ha grid with 9 sampling points
spaced 50 m apart. Soil blocks of 25 25 cm to 10 cm depth were collected from 0 to 10 cm, 1020 cm, and 2030 cm soil layers at each sampling
point, and sorted according to the standard Tropical Soil Biology and Fertility Institute (TSBF) soil monolith method (Anderson and Ingram, 1993;
Moreira et al., 2008). In total 81 monoliths were excavated. The animals
were carefully hand-sorted from the soil blocks in a large tray, immediately after the sampling procedure. Operationally macrofauna have been dened here as the groups in which its individuals are visible to the naked
eye (Kevan, 1968). Organisms from the litter were added with the
010 cm soil macrofauna. The earthworms were preserved in 92.8% ethanol and all the others individuals in 70% ethanol for subsequent laboratory identication and counting. The invertebrates were sorted into the
taxonomic groups: Aranae, Blattodea, Chilopoda, Coleoptera, Dermaptera,
Diplopoda, Diptera, Formicidae, others Hymenoptera, Gastropoda,
Hemiptera, Isopoda, Isoptera, Oligochaeta, and Scorpiones.
Samples for soil chemical characterization were taken at the same
points and time of the macrofauna data, and these data were used to

verify correlations between soil chemical attributes and soil macrofauna. Within a radius of 6 m around each central monolith used
for macrofauna extraction, 12 subsamples were collected from 0 to
10 cm, 1020 cm, and 2030 cm soil layers using a soil Dutch
auger, and composited for subsequent analyses. Soil chemical attributes were measured by analytical methods described in van Raij
et al. (2001): active acidity (pHCaCl2 0.01 mol L 1), potential acidity
(H + Al) by SMP solution, available calcium (Ca) and magnesium
(Mg) by ion-exchange resin/atomic absorption, potassium (K) by
ion-exchange resin/atomic emission, phosphorus (P) by ionexchange resin/colorimetry, and sulfur (Ssulphate) by calcium
phosphate/turbidimetry. SOC and nitrogen (N) were determined by
dry combustion on elemental analyzer LECO CN-2000 (furnace
at 1350 C in pure oxygen).
2.3. Data analysis
The faunal abundance was calculated as the number of individuals
per surface unit (m2). Faunal diversity was assessed by richness (number of groups), and the Shannon's diversity index (H), and evenness
(E). H is calculated by the formula:

H  pi log2 pi:
i1

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

163

Table 1
Climate, soil classication according to Soil Survey Staff (2014), and soil chemical characterization for the study sites.
Site

MAPa Land useb Soil classication

Soil layer pH CaCl2 H + Al

mm

(cm)

Lat_17S 1600

NV

PA

SC

Lat_21S 1240

NV

PA

SC

Lat_23S 1470

NV

PA

SC

a
b
c

Anionic Acrudox

010
1020
2030
Typic Hapludox
010
1020
2030
Anionic Acrudox
010
1020
2030
Typic Rhodudalf
010
1020
2030
Typic Kandiudult
010
1020
2030
Typic Hapludalf
010
1020
2030
Rhodic Hapludox 010
1020
2030
Rhodic Kandiudox 010
1020
2030
Rhodic Hapludox 010
1020
2030

Ca

Mg

SOCc

(mmolc dm3) (mmolc dm3) (mmolc dm3) (g kg1) (mmolc dm3) (mg dm3) (g kg1)
3.8
3.8
3.9
3.7
3.8
3.8
5.1
5.1
4.8
6.5
6.3
6.3
4.0
3.9
4.0
5.3
5.1
4.7
3.7
3.7
3.7
4.7
4.5
4.5
5.5
5.3
5.4

92.8
78.1
64.8
55.1
46.8
39.1
30.0
30.1
33.1
14.7
14.2
13.6
48.3
44.5
39.0
25.2
26.8
31.7
152.8
156.6
152.7
52.5
55.8
53.7
29.7
35.9
33.6

4.2
2.6
2.1
3.0
2.2
2.3
21.7
20.2
15.7
101.6
82.5
69.2
8.3
6.3
8.0
39.0
29.8
24.2
17.1
15.3
12.9
32.6
29.6
24.6
47.5
48.8
47.1

3.4
2.3
1.8
1.6
1.1
1.2
9.3
8.9
6.9
18.6
16.7
14.2
5.2
3.5
3.5
15.6
12.4
9.5
9.1
8.1
7.4
19.9
16.2
12.3
20.1
19.3
18.3

1.2
1.0
0.9
0.7
0.5
0.5
0.8
0.7
0.6
2.2
1.7
1.6
1.1
0.8
0.6
1.1
1.0
0.8
3.1
3.0
2.6
2.6
2.1
1.6
1.5
1.4
1.5

1.0
0.8
0.7
0.6
0.5
0.5
0.6
0.6
0.5
2.8
2.9
2.4
4.2
3.4
3.1
3.1
2.9
2.6
2.8
2.4
2.3
4.4
4.5
4.1
3.1
2.3
2.0

5.6
4.5
3.5
3.0
2.6
2.5
7.3
7.0
4.7
17.3
12.5
9.9
7.1
3.9
3.2
19.6
13.2
7.8
14.3
12.4
11.0
11.5
9.8
7.6
8.9
8.6
7.2

15.6
12.9
10.7
9.5
8.4
6.4
10.8
10.4
9.7
21.8
16.0
14.9
13.3
9.5
7.5
11.0
9.9
8.0
36.7
33.7
30.3
36.4
27.6
20.6
18.9
18.4
17.1

MAP = mean annual precipitation Obtained from CIIAGRO and CEPAGRI climate data from Jata, GO, Valparaso, SP, and Ipaussu, SP.
NV = native vegetation; PA = pasture; SC = sugarcane crop.
SOC = soil organic carbon.

where pi = probability of meeting a taxon I on a plot and s = total

number of taxa encountered on the plot. H is at a maximum when all
taxa are of equal abundance and is 0 when there is only one taxon.
The evenness index is a ratio between the calculated diversity and the
theoretical maximum diversity:

H0
log2 s

E represents the distribution of taxa and assesses the balance of populations. E is equal to 1 when all taxa are of equal abundance and tends
to 0 when one or a few taxa largely dominate the community.
We performed a linear mixed effects analysis of the relationship between individual macrofauna variables (abundance, groups richness,
Shannon index, and evenness) and land use. As xed effects, we entered
land use and soil layer depth (without interaction term) into the model.
As random effects, we had intercepts for sites, as well as by-sites
random slopes for the effect of the macrofauna variable analyzed. Pvalues were obtained by likelihood ratio tests of the full model with
the effect in question against the model without the effect in question.
Macrofauna communities were ordinated with two methods: rst,
land uses were ordinated by their dissimilarity in groups composition
with non-metric multidimensional scaling (NMDS) on Bray-Curtis dissimilarity matrix of macrofauna groups abundance data to reduce dimensionality and allow the visualization of major patterns structuring
the community, followed by non-parametric multivariate analysis of
variance (npMANOVA) to test the effect of LUC and site; secondly, a
principal component analysis (PCA) was performed to visualize the correlations among the abundance of dominant macrofauna groups
(Isoptera, Oligochaeta, Coleoptera and Formicidae) and soil acidity, contents of SOC and contents of macronutrients. All analyses were conducted using the software R, version 3.2.2 (Team RDC, 2014), and we ran the
packages vegan (Dixon, 2003), lme4 (Bates et al., 2015), and ggplot2
(Wickham, 2009).

3. Results
Samples corresponding to native vegetation, pasture, and sugarcane
appeared separated on the NMDS plot (Fig. 2), suggesting that the abundance of the macrofauna groups varied among land uses. While most
groups were associated with samples taken in native vegetation, highly
abundant Isoptera and Oligochaeta appeared associated with pasture
soils (Fig. 2). Differences in community composition among land uses
were conrmed by npMANOVA (df = 2, F = 6.11, r2 = 0.12, p =
0.01) which also demonstrated a signicant effect of site (df = 1, F =
4.35, r2 = 0.04, p = 0.01), and land use site (df = 2, F = 2.91, r2 =
0.06, p = 0.01) on the abundances of the macrofauna groups.
Linear mixed effects analysis showed that land use affected total
abundance of macroinvertebrates (X2(1) = 6.79, p = 0.03), increasing

Fig. 2. Non-parametric multidimensional scaling (NMDS) performed abundances of the

macrofauna groups sampled, with samples grouped by land use. Abbreviations: Aran =
Aranae, Blat = Blattodea, Chilo = Chilopoda, Col = Coleoptera, Derm = Dermaptera,
Diplo = Diplopoda, Dipt = Diptera, Form = Formicidae, Hym = Hymenoptera, Gast =
Gastropoda, Hemp = Hemiptera, Isopo = Isopoda, Isopt = Isoptera, Olig = Oligochaeta,
and Scor = Scorpiones.

164

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

it by about 233.3 individual m2 86.4 (standard error) in 10 cm deep

soil layers from native vegetation to pasture, and lowering it by about
101.5 61.9 from native vegetation to sugarcane. The overall soil macrofauna abundance across all eld sites for 030 cm soil layer ranged
from 411 70 individuals m 2 in native vegetation to 1111
202 individuals m2 in pasture and 106 24 individuals m2 in sugarcane soils. The reduction in the size of the total community from pasture
to sugarcane amounted to 98% at Lat_17S, 78% at Lat_21S, and 90% at
Lat_23S (Fig. 3a).
A total of 14 soil macrofauna taxa were identied. All the 14 taxa occurred in native vegetation, while 11 occurred in pasture, and 9 in sugarcane soils. The native vegetation at Lat_21S was the individual site
with the largest mean number of taxa, 6.4 0.7, and sugarcane at
Lat_17S had the lowest mean, 1.7 0.2 (Table 2). Land use affected
taxa richness in 10 cm soil layers (X2(1) = 6.08, p = 0.04), decreasing
it by 0.13 individual m2 0.38 in from native vegetation to pasture,
and by 1.16 0.40 from native vegetation to sugarcane. The overall reduction in the taxonomic richness for 030 cm soil layer amounted to
24% from native vegetation to pasture, and 39% from pasture to sugarcane, i.e. a mean loss of 53% of the taxonomic richness with the complete land use sequence (Fig. 3b).
Diptera, Blattodea, Hemiptera and Gastropoda were groups exclusively found in native vegetation soils at all sites. More than 90% of the
termites occurred in pasture soils, and the groups Oligocheata,
Dermaptera and Coleoptera were preferably found under pasture as
well. Diplopoda was the single group with the majority of its individuals
found in sugarcane soils (Table 2).
Termites and ants were the most abundant taxa, comprising 43% and
21% of individuals identied, respectively. Termites comprised an average of 23% of the individuals in native vegetation, 58% of the individuals
in pasture, and only 1% in sugarcane soils. Ants accounted for an average
of 26% of the individuals under native vegetation, 5% under pasture, and
18% under sugarcane. Coleoptera comprised 14% of the individuals
identied, and its proportion on the communities increased with the

complete land use sequence (13% of the individuals under native vegetation, 17% under pasture, and 21% in sugarcane soils). Earthworms
accounted for 10% of the total individuals, and were proportionally
more abundant under anthropogenic land uses (17% of the individuals
under pasture and 12% of the individuals under sugarcane) than under
native vegetation (3%) (Table 2).
LUC did not affect Shannon index (H) (X2(1) = 3.55, p = 0.17)
(Fig. 3c). The overall H dropped from 1.6 0.2 in native vegetation to
1.0 0.2 in pasture, and to 0.7 0.2 in sugarcane soils. Although
there were no statistically signicant effects of LUC on evenness (E)
(X2(1) = 3.89, p = 0.14), there was a tendency for one or a few taxa
to dominate the community after LUC (Fig. 3d). The smallest value for
E (0.3 0.1) was found under pasture at Lat_17S.
The top layer 010 cm of the soil pedon harbored on average 69% of
the organisms. Under sugarcane an average of 87% of the organisms
were found in the 010 cm layer, while this value was 80% in native vegetation and 63% in pasture. Overall, macrofauna abundance decreased
by about 245.3 individual m2 59.74 from 0 to 10 to 1020 cm, and
by 275.3 59.74 from 0 to 10 to 2030 cm soil layer. Richness lowered
by about 1.68 groups 0.20 from 0 to 10 to 1020 cm soil layer, and by
1.95 groups 0.20 from 0 to 10 to 2030 cm soil layer. Despite substantially altering the abundance and taxonomic richness of the macrofauna
community, LUC did not change the vertical distribution based on the
proportion of taxa found in the different soil layers; there was a rate
of 2:1 between the number of taxa in the 010 cm layer and the number
of taxa in both 1020 cm and 2030 cm layers at all land uses (Fig. 4).
Fig. 5 shows the distribution of the sampling points colored according to the land use, with soil variables as explanatory variables. Together, the two axes explained 52.7% of the variance. In general, pasture soils
(green) were grouped near the macrofauna variables, while sugarcane
soils (blue) were mainly associated with higher pH and higher contents
of Ca and Mg. Native vegetation soils (red) were widely distributed.
Isoptera and Oligochaeta showed a weak negative correlation with pH.
The abundance of Isoptera, Oligochaeta and Coleoptera was negatively

Fig. 3. Soil macrofauna abundance (a), taxonomic richness (b), diversity (c), and taxa distribution (d) in the 030 cm soil layer as a function of the land use change. n = 9, error bars denote
standard error of the mean.

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

165

Table 2
Abundance (individuals m2) and standard error of soil macrofuna groups in the 030 cm layer. n = 9.
Taxa

Common name

Taxonomic level

Lat_17S

Lat_21S

Lat_23S

NV

PA

SC

NV

PA

SC

NV

PA

SC

Earthworms
Oligochaeta

Earthworms

Subclass

4 (2.4)

9 (5.4)

4 (2.4)

21 (9.2)

373 (102.9)

25 (7.1)

9 (3.9)

60 (15.5)

11 (7.1)

Insects
Isoptera
Hymenoptera
Formicidae
Coleoptera
Blattodea
Dermaptera
Diptera
Hemiptera

Termites
Oth. than ants
Ants
Beetles
Cockroaches
Earwigs
Mosquitoes
Bugs

Order
Order
Family
Order
Order
Order
Order
Order

66 (26.7)
9 (3.9)
4 (2.4)
25 (10.7)
0
0
0
0

1093 (309.2)
0
55 (21.5)
308 (277.7)
0
0
0
0

0
4 (2.4)
0
11 (4.6)
0
2 (1.8)
0
0

11 (7.1)
7 (3.9)
144 (70.0)
82 (30.5)
0
11 (6.0)
12 (7.4)
2 (1.8)

78 (66.4)
2 (1.8)
78 (37.4)
247 (64.2)
0
16 (7.1)
0
0

0
2 (1.8)
2 (1.8)
21 (13.6)
0
2 (1.8)
0
0

151 (72.4)
0
272 (102.9)
25 (10.4)
2 (1.8)
0
5 (5.3)
0

924 (488.5)
0
14 (5.6)
5 (2.7)
0
0
0
0

2 (1.8)
0
55 (30.4)
20 (10.6)
0
0
0
0

Myriapods
Diplopoda
Chilopoda

Millipedes
Centipedes

Class
Class

4 (2.4)
2 (1.8)

0
7 (2.8)

14 (4.2)
0

44 (25.8)
16 (8.4)

0
9 (6.0)

92 (38)
36 (35.6)

32 (6.0)
82 (24.2)

30 (12.9)
5 (2.7)

7 (4.7)
9 (4.7)

Arachnids
Araneae
Scorpiones

Spiders
Scorpions

Order
Order

0
0

2 (1.8)
2 (1.8)

0
0

25 (7.6)
0

2 (1.8)
0

0
0

30 (11.8)
5 (5.3)

2 (1.8)
0

2 (1.8)
0

Molluscs
Gastropoda

Snails

Class

2 (1.8)

4 (2.4)

5 (3.8)

Crustaceans
Isopoda

Woodlouse

Order

37 (37.3)

84 (64.4)

9 (7.1)

correlated to the contents of P and K. Isoptera was closed correlated to

H + Al, and Fomicidae was correlated to higher contents of SOC, N
and S (Fig. 5).
4. Discussion
Experimental and observational evidence is emerging that LUC associated with bioenergy production in the tropics aggravate biodiversity
loss (Desiree et al., 2014; Koh and Wilcove, 2008). However, most of
these studies assessing bioenergy-cropping impacts on biodiversity examined plants and aboveground animals as biodiversity indicators.
Much less is known about the responses of the belowground communities to bioenergy crop expansion. Thus, the present study extends this
body of research through the assessment of soil fauna responses to the
sugarcane expansion over pasturelands in Brazil. We conducted identical observational studies in three chronosequences along two major
tropical biomes to assess the impact of replacing pastures with sugarcane crops on soil macrofauna. We also assessed whether the abundance of soil macrofauna is related to soil chemical status.
4.1. Land use change and macrofauna abundance and diversity
Native vegetation forests were sampled at the three sites because
typical LUC sequences in the tropics start with the conversion of native forest into pasture for cattle ranching and later the conversion of
pasture into cropland, but also to provide a reference point for the
macrofauna community at the native ecosystems. As expected, native vegetation soils were highly diverse in macrofauna groups, suggesting higher complement of groups and its ecological functions
compared to pasture and sugarcane soils. In line with this is the
fact that higher trophic level groups were more abundant in native
forest soils; greater abundance of predatory groups such as Araneae
and Scorpions under native vegetation than in pasture and sugarcane soils. Ants and termites were the most abundant groups at native forests, same pattern described in previous studies in Cerrado
(Silva et al., 2007; Batista et al., 2014). Both ants and termites play
a key role in the function of tropical ecosystems (Lavelle et al.,
2006), for instance by forming and stabilizing the structure of

heavily weathered soils (Ayuke et al., 2011). Field observations

showed a strong soil microgranular structure under native forests
at all three sites, which in tropical soils is mainly related to soil bioturbation by termites (Jungerius et al., 1999; Reatto et al., 2009).
Contrary to annual crops, pastures cultivated from native forests in
tropical ecosystems generally provide suitable soil habitats for a high
abundance of specic soil macrofauna groups (Decaens et al., 2004).
Our study conrms other studies that show the size of the macrofaunal
community in tropical soils tends to increase over time after conversion
of native vegetation to pasture (Benito et al., 2004; Black and Okwakol,
1997; Decaens et al., 2004; Lavelle et al., 2014). Increases in the
abundance of only two groups drove this pattern: average termite
abundance increased ninefold and earthworm densities increased
thirteenfold from native vegetation to pasture, and these two groups
largely dominated the community under pasture (Table 2). The standard error for termite abundance in native forest soils was higher than
in the other land uses, which suggests a more heterogeneous spatial distribution of these animals. Increased C inputs from cattle dung return in
pasture systems make a large contribution to support greater densities
of earthworms (Schon et al., 2015) and termites (Freymann et al.,
2008). While not examined here, this dominance of earthworms and
termites in pasture soils may have positive impacts in the structural stability of tropical soils (Jouquet et al., 2006), as these animals have the
ability to move through the soil building stable biogenic structures
with specic physical, chemical and microbiological properties
(Jouquet et al., 2006). Yet, the recycling of cattle dung by termites and
earthworms in pasturelands enhances C incorporation and stabilization
into soil (Freymann et al., 2008; Schon et al., 2015). Although the macrofauna community was more abundant in pasture compared to native
forest, the diversity of macrofauna groups was lower, including losses of
predatory groups (Araneae and Scorpiones). This result is certainly inuenced by the lower plant diversity in pasture systems, as the positive
relationship between plant diversity and soil biodiversity is well described (Hooper et al., 2005; van der Heijden et al., 2008). Yet, the conversion of native forests to pastures in our study sites involved clearing
native vegetation through the slash-and-burn technique, which potentially impacted all the living forms in soil. Thus pasture introduction in
tropical ecosystems favors a few groups that can consume the new

166

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

Fig. 4. Vertical distribution and taxonomic group composition of soil macrofauna within soil pedon under native vegetation, pasture, and sugarcane at three eld sites. Numbers in italics
refer to total density in each layer. n = 9. Abbreviations: Aran = Aranae, Chi = Chilopoda, Col = Coleoptera, Dip = Diplopoda, Form = Formicidae, Isop = Isopoda, Isopt = Isoptera, and
Olig = Oligochaeta.

organic resource available and interact within an unbalanced food web

where predation becomes less inuential.
In general, the conversion of pasture to sugarcane negatively impacted the abundance, taxonomic richness and H of the soil macrofaunal community (Fig. 3a,b & c). Sugarcane soils as well as the majority of
croplands are managed at moderate to strong intensity, with deep tillage and plowing, and systematic use of chemical inputs, such as pesticides and fertilizers (Bell et al., 2007). Despite the proportionally low
number of agricultural pests in these soils, pesticides are commonly
used, with negative impacts on entire belowground community. In the
last decade a shift from manual to mechanical harvesting in Brazil has
caused the rapid proliferation and spread of certain sugarcane pests
(e.g. root spittlebug, curculionid, sugarcane borer) (Dinardo-Miranda
and Fracasso, 2013). Consequently, the application of chemicals is increasing, as it is still the more efcient alternative of control against
these pests. The intensive use of mineral fertilizers as observed in sugarcane soils negatively affects the abundance, diversity and equability of
soil macrofaunal community (Alves et al., 2008; Giller et al., 1997;
Sileshi and Mafongoya, 2006). At the sampling time the sugarcane
areas were in different phases of its 5-year rotation period: sugarcane
was in the third ratoon cropping at Lat_17S, fourth ratoon cropping at
Lat_21S, and fth ratoon cropping at Lat_23S (Table A.1). This makes a
good t with ants abundance data: no ants were found at Lat_17S,
2 1.8 individuals m 2 were found at Lat_21S, and 55

30.4 individuals m2 were found at Lat_23S (Table 2). These ndings

may indicate that: (i) ants are completely lost after the cultivation operations and elds need to be recolonized by founding-queens; and (ii)
the 5-years interval between cultivation cycles in sugarcane elds
may be long enough to provide conditions for ants recolonization in
the nal years of the rotation cycle. The reduced number of individuals
and groups of macrofauna in sugarcane soils are expected to affect different levels of the soil food web. For instance, soil fauna make litter
and the products of its physical and chemical degradation more available to soil microbes (Petersen and Luxton, 1982; Chamberlain et al.,
2006), and the selective removal of larger invertebrates from soil is
demonstrated to affect microbes reducing the size of the micro-food
web, the ultimate actors in the decomposition process, with implications for nutrient availability and temporal dynamics (Carrillo et al.,
2011).
The concentration of macroinvertebrates in the upper 10 cm of soil in
this study (Fig. 4) was higher than in other studies (Barros et al., 2003;
Pauli et al., 2011), and unlike these studies, it seems disconnected with
the vertical variations in the contents of SOC (Table 1). Sugarcane soils,
for example, appeared to have SOC homogenization in the upper 30 cm
soil (Table 1) as a result of tillage operations: nevertheless 87% of the organisms were found in the upper 10 cm of soil (Fig. 4). The vertical distribution is likely a function of the litter concentration at surface and the
lower bulk density in the 010 cm soil layer (Cherubin et al., 2015).

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

167

Fig. 5. Principal component analysis of the abundance of major macrofauna groups (Oligochaeta, Isoptera, Coleoptera, and Formicidae) and soil chemical attributes (pH CaCl2 (pH),
exchangeable aluminum (H + Al), calcium (Ca), magnesium (Mg), potassium (K), phosphorus (P), nitrogen (N), soil organic carbon (SOC), of 243 sampling points across three land
uses: native vegetation (red), pasture (green), and sugarcane (blue).

4.2. Relationships between macrofauna and soil chemical attributes

PCA indicated the preferential occurrence of the four most abundant
macrofauna groups in pasture soils rather than sugarcane soils (Fig. 5).
As discussed above, conversion of pasture to sugarcane as well as sugarcane crop management involves tillage that destroys the habitats of soil
invertebrates. In this study, higher densities of macrofauna animals
were specially related to the wide dominance of termites in the communities of pasture soils at Lat_17S and Lat_23S (Table 2).
The positive relationship between soil acidity and macrofauna abundance is illustrated in the PCA by the negative correlations between pH
and the abundance of the major macrofauna groups, and the consequent positive correlations between higher contents of H + Al and macrofauna abundance (Fig. 5). But instead of ecological reasons, the
association of more acid soils and higher abundance of macrofauna
can be explained by the soil management: the absence of lime application in pasture soils (Cherubin et al., 2015), where macrofauna were
more abundant, in contrast with the high inputs of lime to neutralize
soil acidity in soils under sugarcane. The successive applications of
lime and fertilizers in sugarcane soils also explain the association of sugarcane soils with higher contents of Ca, Mg, P and K, and the general
negative correlations between these macronutrients and macrofauna
abundance.
Although the expansion of sugarcane ethanol in Brazil delivers substantial greenhouse-gas emissions savings when displacing fossil based
fuels (Egeskog et al., 2014; Mello et al., 2014), our results draw attention
for a signicant biodiversity loss due to sugarcane expansion from
pasturelands and warrant efforts to understand the impacts of altering
belowground biodiversity and composition on the performance
of agroecosystems. Further research is needed to link these soil

macrofauna responses to sugarcane expansion and changes in others

soil quality indicators, and to assess management practices that potentially can minimize the detrimental effects of sugarcane cropping on
soil macrofauna. How these soil macrofauna responses to LUC in sugarcane expansion areas alter the complex food web structure in which soil
organisms interact? Yet how detrimental are these modications in soil
macrofaunal community for the delivery of ecosystem services provided by soil biota and benecial purposes to agriculture? These are key issues to be claried in future researches.
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.scitotenv.2016.04.116.
Acknowledgements
A.L.C.F. and M.R.C. thank So Paulo Research Foundation FAPESP
(2012/22510-8 and 2013/17581-6) for the scholarships granted while
this research was carried out. We thank Bruno Castigioni, Evandro
Schonell, Eduardo Lucianer, Edpool Rocha, Manuela Testa, and Lucas
Canisares for lab and eld assistance. We are grateful for two anonymous referees for many helpful comments.
References
Adami, M., Rudorff, B.F.T., Freitas, R.M., Aguiar, D.A., Sugawara, L.M., Mello, M.P., 2012.
Remote sensing time series to evaluate direct land use change of recent expanded
sugarcane crop in Brazil. Sustainability 4, 574585.
Alves, M.V., Santos, J.C.P., de Gois, D.T., Alberton, J.V., Baretta, D., 2008. Soil macrofauna as
inuenced by chemical fertilizers and swine manure use in western Santa Catarina
state, Brazil. Rev. Bras. Cienc. Solo 32, 589598.
Anderson, J.M., Ingram, J.S.I., 1993. Tropical Soil Biology and Fertility: A Handbook of
Methods. CAB International, Wallingford.

168

A.L.C. Franco et al. / Science of the Total Environment 563564 (2016) 160168

Ayuke, F.O., Brussaard, L., Vanlauwe, B., Six, J., Lelei, D.K., Kibunja, C.N., Pulleman, M.M.,
2011. Soil fertility management: impacts on soil macrofauna, soil aggregation and
soil organic matter allocation. Appl. Soil Ecol. 48, 5362.
Barros, E., Neves, A., Blanchart, E., Fernandes, E.C.M., Wandelli, E., Lavelle, P., 2003. Development of the soil macrofauna community under silvopastoral and agrosilvicultural
systems in Amazonia. Pedobiologia 47, 273280.
Bates, D., Mchler, M., Bolker, B.M., Walker, S.C., 2015. Fitting linear mixed-effects models
using lme4. J. Stat. Softw. 67, 148.
Batista, I., Correia, M.E.F., Pereira, M.G., Bieluczyk, W., Schiavo, J.A., Rouws, J.R.C., 2014. Oxidizable fractions of total organic carbon and soil macrofauna in a crop-livestock integration system. Rev. Bras. Cinc. Solo 38, 797809.
Bell, M.J., Stirling, G.R., Pankhurst, C.E., 2007. Management impacts on health of soils
supporting Australian grain and sugarcane industries. Soil Tillage Res. 97, 256271.
Benito, N.P., Brossard, M., Pasini, A., Guimaraes, M.D., Bobillier, B., 2004. Transformations
of soil macroinvertebrate populations after native vegetation conversion to pasture
cultivation (Brazilian Cerrado). Eur. J. Soil Biol. 40, 147154.
Black, H.I.J., Okwakol, M.J.N., 1997. Agricultural intensication, soil biodiversity and
agroecosystem function in the tropics: the role of termites. Appl. Soil Ecol. 6, 3753.
Brussaard, L., de Ruiter, P.C., Brown, G.G., 2007. Soil biodiversity for agricultural sustainability. Agric. Ecosyst. Environ. 121, 233244.
Callaham, M.A., Richter, D.D., Coleman, D.C., Hofmockel, M., 2006. Long-term land-use effects on soil invertebrate communities in southern piedmont soils, USA. Eur. J. Soil
Biol. 42, S150S156.
Carrillo, Y., Ball, B.A., Bradford, M.A., Jordan, C.F., Molina, M., 2011. Soil fauna alter the effects of litter composition on nitrogen cycling in a mineral soil. Soil Biol. Biochem. 43,
14401449.
Castellanos-Navarrete, A., Rodriguez-Aragones, C., de Goede, R.G.M., Kooistra, M.J., Sayre,
K.D., Brussaard, L., et al., 2012. Earthworm activity and soil structural changes under
conservation agriculture in Central Mexico. Soil Tillage Res. 123, 6170.
Chamberlain, P.M., McNamara, N.P., Chaplow, J., Stott, A.W., Black, H.I.J., 2006. Translocation of surface litter carbon into soil by Collembola. Soil Biol. Biochem. 38, 26552664.
Cherubin, M.R., Franco, A.L.C., Cerri, C.E.P., Oliveira, D.M.D., Davies, C.A., Cerri, C.C., 2015.
Sugarcane expansion in Brazilian tropical soils-effects of land use change on soil
chemical attributes. Agric. Ecosyst. Environ. 211, 173184.
Dauber, J., Bolte, A., 2014. Bioenergy: challenge or support for the conservation of biodiversity? Glob. Chang. Biol. Bioenergy 6, 180182.
de Oliveira, F.S., Varajao, A.F.D.C., Varajao, C.A.C., Schaefer, C.E.G.R., Boulange, B., 2014. The
role of biological agents in the microstructural and mineralogical transformations in
aluminium lateritic deposit in Central Brazil. Geoderma 226, 250259.
de Vries, F.T., Thebault, E., Liiri, M., Birkhofer, K., Tsiafouli, M.A., Bjornlund, L., et al., 2013.
Soil food web properties explain ecosystem services across European land use systems. Proc. Natl. Acad. Sci. U. S. A. 110, 1429614301.
Decaens, T., Jimenez, J.J., Barros, E., Chauvel, A., Blanchart, E., Fragoso, C., et al., 2004. Soil
macrofaunal communities in permanent pastures derived from tropical forest or savanna. Agric. Ecosyst. Environ. 103, 301312.
Decaens, T., Jimenez, J.J., Gioia, C., Measey, G.J., Lavelle, P., 2006. The values of soil animals
for conservation biology. Eur. J. Soil Biol. 42, S23S38.
Desiree, J.I., Pita, A.V., Floor, V.D.H., Andre, P.C.F., 2014. Biodiversity impacts of bioenergy
crop production: a state-of-the-art review. Glob. Chang. Biol. Bioenergy 6, 183209.
Dinardo-Miranda, L.L., Fracasso, J.V., 2013. Sugarcane straw and the populations of pests
and nematodes. Sci. Agric. 70, 369374.
Dixon, P., 2003. VEGAN, a package of R functions for community ecology. J. Veg. Sci. 14,
927930.
Egeskog, A., Freitas, F., Berndes, G., Sparouek, G., Wirsenius, S., 2014. Greenhouse gas balances and land use changes associated with the planned expansion (to 2020) of the
sugarcane ethanol industry in Sao Paulo, Brazil. Biomass Bioenergy 63, 280290.
Franco, A.L.C., Cherubin, M.R., Pavinato, P.S., Cerri, C.E.P., Six, J., Davies, C.A., et al., 2015.
Soil carbon, nitrogen and phosphorus changes under sugarcane expansion in Brazil.
Sci. Total Environ. 515, 3038.
Freymann, B.P., Buitenwerf, R., de Souza, O., Olff, H., 2008. The importance of termites
(Isoptera) for the recycling of herbivore dung in tropical ecosystems. Eur.
J. Entomol. 105, 165173.
Giller, K.E., Beare, M.H., Lavelle, P., Izac, A.M.N., Swift, M.J., 1997. Agricultural intensication, soil biodiversity and agroecosystem function. Appl. Soil Ecol. 6, 316.
Goldemberg, J., Coelho, S.T., Guardabassi, P., 2008. The sustainability of ethanol production from sugarcane. Energ Policy 36, 20862097.
Goldemberg, J., Mello, F.F.C., Cerri, C.E.P., Davies, C.A., Cerri, C.C., 2014. Meeting the global
demand for biofuels in 2021 through sustainable land use change policy. Energ Policy
69, 1418.
Hernandez-Ruiz, P., Castano-Meneses, G., 2006. Ants (hymenoptera: Formicidae) diversity in agricultural ecosystems at Mezquital Valley, hidalgo, Mexico. Eur. J. Soil Biol. 42,
S208S212.
Hooper, D.U., FS III, Chapin, Ewel, J.J., Hector, A., Inchausti, P., Lavorel, S., Lawton, J.H.,
Lodge, D.M., Loreau, M., Naeem, S., Schmid, B., 2005. Effects of biodiversity on ecosystem functioning: a consensus of current knowledge. Ecol. Monogr. 75, 335.

Jouquet, P., Dauber, J., Lagerlof, J., Lavelle, P., Lepage, M., 2006. Soil invertebrates as ecosystem engineers: intended and accidental effects on soil and feedback loops. Appl. Soil
Ecol. 32, 153164.
Jungerius, P.D., van den Ancker, J.A.M., Mucher, H.J., 1999. The contribution of termites to
the microgranular structure of soils on the Uasin Gishu plateau, Kenya. Catena 34,
349363.
Kevan, DKMcE, 1968. Soil animals. Gateshead on Tyne. Northumberland Press, UK.
Koh, L.P., Wilcove, D.S., 2008. Is oil palm agriculture really destroying tropical biodiversity? Conserv. Lett. 1, 6064.
Lavelle, P., Decaens, T., Aubert, M., Barot, S., Blouin, M., Bureau, F., et al., 2006. Soil invertebrates and ecosystem services. Eur. J. Soil Biol. 42, S3S15.
Lavelle, P., Rodriguez, N., Arguello, O., Bernal, J., Botero, C., Chaparro, P., et al., 2014. Soil
ecosystem services and land use in the rapidly changing Orinoco River basin of
Colombia. Agric. Ecosyst. Environ. 185, 106117.
Leonard, J., Rajot, J.L., 2001. Inuence of termites on runoff and inltration: quantication
and analysis. Geoderma 104, 1740.
Lubbers, I.M., van Groenigen, K.J., Fonte, S.J., Six, J., Brussaard, L., van Groenigen, J.W., 2013.
Greenhouse-gas emissions from soils increased by earthworms. Nat. Clim. Chang. 3,
187194.
Majeed, M.Z., Miambi, E., Barois, I., Randriamanantsoa, R., Blanchart, E., Brauman, A., 2014.
Contribution of white grubs (Scarabaeidae: Coleoptera) to N2O emissions from tropical soils. Soil Biol. Biochem. 75, 3744.
McCormack, S.A., Ostle, N., Bardgett, R.D., Hopkins, D.W., Vanbergen, A.J., 2013. Biochar in
bioenergy cropping systems: impacts on soil faunal communities and linked ecosystem processes. Glob. Chang. Biol. Bioenergy 5, 8195.
Mello, F.F.C., Cerri, C.E.P., Davies, C.A., Holbrook, N.M., Paustian, K., Maia, S.M.F., et al.,
2014. Payback time for soil carbon and sugar-cane ethanol. Nat. Clim. Chang. 4,
605609.
Moreira, F.M.S., Huising, E.J., Bignell, D., 2008. A Handbook of Tropical Soil Biology: Sampling and Characterization of Below-ground Biodiversity. Earthscan, London.
Neto, F.V.D., Correia, M.E.F., Pereira, G.H.A., Pereira, M.G., Leles, P.S.D., 2012. Soil Fauna as
an indicator of soil quality in Forest stands, pasture and secondary Forest. Rev. Bras.
Cienc. Solo 36, 14071417.
Pauli, N., Barrios, E., Conacher, A.J., Oberthur, T., 2011. Soil macrofauna in agricultural
landscapes dominated by the Quesungual slash-and-mulch agroforestry system,
Western Honduras. Appl. Soil Ecol. 47, 119132.
Petersen, H., Luxton, M., 1982. A comparative analysis of soil fauna populations and their
role in decomposition processes. Oikos 39, 287388.
Reatto, A., Bruand, A., Martins, E.D., Muller, F., da Silva, E.M., de Carvalho, O.A., Brossard,
M., Richard, G., 2009. Development and origin of the microgranular structure in latosols of the Brazilian central plateau: signicance of texture, mineralogy, and biological activity. Catena 76, 122134.
Schon, N.L., Mackay, A.D., Gray, R.A., Dodd, M.B., van Koten, C., 2015. Quantifying dung
carbon incorporation by earthworms in pasture soils. Eur. J. Soil Sci. 66, 348358.
Sileshi, G., Mafongoya, P.L., 2006. Long-term effects of improved legume fallows on soil invertebrate macrofauna and maize yield in Eastern Zambia. Agric. Ecosyst. Environ.
115, 6978.
Silva, R., Tomazi, M., Pezarico, C.R., Aquino, A.M., Mercante, F.M., 2007. Macrofauna
invertebrada edca em cultivo de mandioca sob sistemas de cobertura do solo.
Pesq. Agrop. Brasileira 42, 865871.
Six, J., Bossuyt, H., Degryze, S., Denef, K., 2004. A history of research on the link between
(micro)aggregates, soil biota, and soil organic matter dynamics. Soil Tillage Res. 79,
731.
Staff SS, 2014. Keys to Soil Taxonomy. USDA - Natural Resources Conservation Service,
Washington, DC.
Swift, M.J., Heal, O.W., Anderson, J.M., 1979. Decomposition in Terrestrial Ecosystems.
University of California Press, Berkely.
Team RDC, 2014. R: A Language and Environment for Statistical Computing (Vienna,
Austria).
Van Der Heijden, M.G.A., Bardgett, R.D., Van Straalen, N.M., 2008. The unseen majority:
soil microbes as drivers of plant diversity and productivity in terrestrial ecosystems.
Ecol. Lett. 11, 296310.
van Raij, B., Andrade, J.C., Cantarella, H., Quaggio, J.A., 2001. Anlise Qumica para
Avaliao da Fertilidade de solos Tropicais. Campinas, Instituto Agronmico.
Verdade, L.M., Gheler-Costa, C., Penteado, M., Dotta, G., 2012. The impacts of sugarcane
expansion on wildlife in the state of So Paulo, Brazil. J. Sustain. Bioenergy Syst. 2,
138144.
Wagg, C., Bender, S.F., Widmer, F., van der Heijden, M.G.A., 2014. Soil biodiversity and soil
community composition determine ecosystem multifunctionality. Proc. Natl. Acad.
Sci. U. S. A. 111, 52665270.
Wickham, H., 2009. ggplot2: Elegant graphics for data analysis. Springer-Verlag, New
York.
Wiens, J., Fargione, J., Hill, J., 2011. Biofuels and biodiversity. Ecol. Appl. 21, 10851095.