Corresponding author: Thais Giovannini Pellegrini (
Academic editor: O. Moldovan
Patterns of biodiversity respond to habitat disturbances and different land-uses. Those patterns possibly vary according to the spatial scale under analysis. Although other studies have shown such responses for different systems, no study has ever demonstrated spatial-scale influences in subterranean terrestrial communities. Therefore, the objective of this paper was to analyze how land use and cave physical structure could influence the terrestrial cave invertebrate species composition. We also determined the influence of different spatial scale on the structure of invertebrate cave composition. We collected environmental data at local scale (e.g. cave size, substrate and environmental stability). For spatial scale we determined land uses at three different landscape scales; we gathered these data into circular areas of different sizes (50, 100 and 250 meters) with centroids in the cave entrances. We finally performed three Distance Based Linear Modeling analyses to test for differences among the predictability of environmental variables when comparing different spatial scales. The best explanatory variable for cave invertebrate similarities was the percentage of covering of the external environment by limestone outcrops. We confirm the scale-dependence hypothesis through the different patterns showed among distinct buffer areas. Models become more precise when larger scales were analyzed to explain cave invertebrate composition. This suggests that larger scales capture important environmental features that explain the cave fauna similarities more precisely. Additionally, we found a strong influence of limestone outcrops at all landscape scale structuring cave communities.
Pellegrini TG, Sales LP, Aguiar P, Ferreira RL (2016) Linking spatial scale dependence of land-use descriptors and invertebrate cave community composition. Subterranean Biology 18: 17–38. doi:
Environmental heterogeneity in natural landscapes has been historically replaced by anthropogenic mosaics around the world. As a consequence, several hypothesis describing how landscape characteristics affect biodiversity patterns have been proposed (
In the context of landscape influences on biodiversity distribution patterns, caves are good models since they represent simplified and fragile ecosystems (
The main goal of this paper is to explain cave invertebrate composition through environmental variables from within the cave and also from the landscape surrounding the caves at different spatial scales. To that end, we tested the hypothesis that the spatial scale affects the predictability of environmental variables. We also checked for grouping patterns among cave fauna according to the most explanatory variables.
The present study was carried out at the conservation unit “
Spatial characterization of landscape at “Parque Estadual do Sumidouro”. Different colors represent distinct vegetation cover or land-use types. The numbers indicate the sampled caves, indicated by name. Legend:
We only used terrestrial invertebrates for our analysis because they account for most of cave richness and abundance in Brazilian caves (
We considered environmental variables at different spatial scales, thus encompassing traits inside the cave (local scale), and those belonging to the landscape scale. At the local scale, we measured the linear extension of the cave and number and size of the entrances. We used those variables to estimate the cave Environmental Stability Index is the linear extension (total length) of the cave is the sum of all entrances extension is the number of entrances is the average distance from all entrances to a reference cave entrance
In order to determine the habitat heterogeneity inside each cave, we classified and quantified the different types of habitat. We divided each cave main conduit into at least 11 transects, equally distanced. Bigger caves were divided with more transects with a maximum distance between them corresponding to 15 meters. Each transect was then subdivided at five points, and at these points we visually examined substrate type (guano, water, trash, organic matter and the size of inorganic grains), along the five equidistant points, encompassing a minimal number of 55 measurements for each cave. The size of inorganic grains was classified into eight classes (bedrock, large boulders, boulders, cobbles, coarse gravel, fine gravel, sand, silt and hardpan). This methodology was modified from
In order to obtain environmental variables at a landscape scale we quantified the main land-use types at different buffers through image classification and matrix characterization. These buffers were circular areas centered at each cave entrance, with a radius of 50 m, 100 m, and 250 m. For those caves with multiple entrances, the biggest entrance was used as reference in this analysis. Therefore, we delimited three circular areas, named respectively Buffer 50 m, Buffer 100 m, and Buffer 250 m (Figure
Spatial characterization of
Study area location, sampling design used in sampled caves at “Parque Estadual do Sumidouro”, and the Buffers of 50m, 100m, and 250m for analyzing the effect of spatial scale on the explanatory power of environmental variables in the cave invertebrate communities.
In order to detect if the geographic distance is responsible for the highest similarities between the studied caves, we performed a Mantel test with PAST 3.11 software ( Distance Based Linear Modeling
We found 186 invertebrate species, distributed along at least 78 families and 23 orders (Table
Taxon and families collected at Parque Estadual do Sumidouro Caves. : Not Identified
TAXON | FAMILIES | Gruta do Grilão | Gruta do Lixo | Gruta do Sumidouro | Gruta Feneme | Gruta Pacas | Helictites | Lagoa Seca | Lapa da Varzea | Macacos da Caverna | Ninho de pérolas | |
---|---|---|---|---|---|---|---|---|---|---|---|---|
|
|
|
0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 |
|
|
|
1 | 0 | 1 | 0 | 2 | 1 | 0 | 1 | 0 | 0 |
|
|
|
0 | 0 | 0 | 0 | 2 | 1 | 0 | 2 | 1 | 1 |
|
|
|
1 | 0 | 1 | 1 | 0 | 1 | 0 | 0 | 1 | 1 |
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
Scolopendromorha |
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
|
|
0 | 1 | 0 | 0 | 1 | 0 | 0 | 1 | 0 | 0 | |
|
|
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 3 | 0 | 0 |
|
0 | 1 | 0 | 1 | 0 | 0 | 1 | 0 | 0 | 0 | ||
|
1 | 1 | 0 | 0 | 0 | 1 | 0 | 1 | 1 | 1 | ||
|
0 | 0 | 0 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | ||
|
0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
|
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | |
|
0 | 0 | 0 | 1 | 0 | 0 | 1 | 0 | 1 | 0 | ||
|
1 | 1 | 2 | 2 | 0 | 1 | 1 | 1 | 2 | 1 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 2 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | ||
|
0 | 0 | 0 | 0 | 1 | 0 | 0 | 1 | 0 | 0 | ||
|
1 | 1 | 2 | 1 | 1 | 2 | 2 | 1 | 1 | 1 | ||
|
0 | 1 | 0 | 0 | 0 | 3 | 2 | 0 | 2 | 1 | ||
|
1 | 1 | 1 | 0 | 0 | 1 | 1 | 1 | 1 | 1 | ||
|
1 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | ||
|
0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | ||
|
0 | 0 | 2 | 0 | 2 | 0 | 0 | 3 | 2 | 0 | ||
|
0 | 1 | 0 | 1 | 0 | 0 | 0 | 1 | 1 | 0 | ||
|
0 | 0 | 0 | 0 | 1 | 1 | 0 | 0 | 0 | 0 | ||
|
|
1 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | 1 | 0 | |
|
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
0 | 0 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | 0 | ||
|
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
|
|
0 | 0 | 1 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | |
|
|
|
0 | 1 | 1 | 0 | 1 | 0 | 0 | 0 | 1 | 0 |
|
|
0 | 0 | 0 | 1 | 0 | 0 | 0 | 2 | 0 | 0 | |
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
|
0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
0 | 0 | 0 | 0 | 1 | 1 | 0 | 0 | 0 | 1 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
|
1 | 2 | 1 | 0 | 0 | 0 | 1 | 2 | 0 | 1 | ||
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
1 | 0 | 0 | 0 | 0 | 1 | 1 | 1 | 0 | 1 | ||
|
|
3 | 0 | 2 | 1 | 2 | 1 | 0 | 4 | 3 | 1 | |
|
|
0 | 1 | 2 | 0 | 1 | 0 | 0 | 2 | 0 | 0 | |
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
0 | 1 | 0 | 1 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
0 | 1 | 0 | 0 | 1 | 1 | 1 | 0 | 1 | 0 | ||
|
1 | 1 | 0 | 0 | 0 | 0 | 1 | 0 | 1 | 0 | ||
|
0 | 1 | 0 | 1 | 0 | 0 | 0 | 3 | 2 | 1 | ||
|
0 | 0 | 0 | 0 | 0 | 1 | 1 | 0 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 1 | 0 | ||
|
0 | 0 | 1 | 1 | 0 | 0 | 0 | 2 | 1 | 0 | ||
|
0 | 0 | 0 | 0 | 1 | 0 | 0 | 1 | 1 | 1 | ||
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 | 1 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | ||
|
|
0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | |
Phahlangopsidae | 1 | 0 | 1 | 2 | 1 | 1 | 0 | 1 | 2 | 1 | ||
|
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 2 | 0 | 0 | |
|
1 | 1 | 0 | 1 | 0 | 1 | 0 | 1 | 1 | 0 | ||
|
0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
Ploiaridae | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | ||
|
1 | 1 | 2 | 0 | 1 | 1 | 1 | 0 | 2 | 1 | ||
|
|
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | |
|
1 | 1 | 1 | 3 | 3 | 3 | 1 | 0 | 2 | 2 | ||
Eulopidae | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
|
0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | |
|
0 | 0 | 0 | 0 | 1 | 0 | 1 | 0 | 0 | 0 | ||
|
|
0 | 1 | 0 | 1 | 0 | 0 | 0 | 1 | 0 | 0 | |
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
1 | 4 | 1 | 3 | 0 | 2 | 0 | 2 | 1 | 2 | ||
|
1 | 2 | 2 | 2 | 1 | 2 | 0 | 2 | 2 | 1 | ||
|
|
0 | 0 | 1 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | |
|
|
2 | 0 | 0 | 1 | 1 | 0 | 0 | 2 | 2 | 1 | |
|
|
2 | 0 | 0 | 1 | 1 | 0 | 0 | 1 | 2 | 1 | |
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
0 | 0 | 1 | 1 | 1 | 0 | 0 | 0 | 1 | 0 | ||
|
0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 1 | 1 | 0 | 0 | 0 | ||
|
0 | 0 | 0 | 0 | 0 | 1 | 1 | 0 | 0 | 0 | ||
|
0 | 1 | 2 | 0 | 1 | 0 | 0 | 0 | 1 | 0 | ||
|
|
0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
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|
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|
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We found no correlation between geographical distance and caves invertebrate similarity (Correlation R MANTEL TEST = -0.3366; p = 0.9111).
Lapa das Pacas Cave had the highest Environmental Stability Index (
Values of physical variables found at each cave at : Environmental Stability Index : Linear Extension : Sum of Entrances Area : Number of Entrances
CAVE |
|
|
|
|
---|---|---|---|---|
|
0.5983 | 42.82 | 2.74 | 3 |
|
0.0829 | 16.85 | 8.28 | 1 |
|
3.3859 | 137.68 | 6.47 | 1 |
|
3.2335 | 26.13 | 0.8 | 1 |
|
1.8935 | 69.48 | 3.45 | 1 |
|
1.7884 | 28.39 | 7.59 | 3 |
|
2.806 | 134.35 | 2.53 | 1 |
|
3.5262 | 319.56 | 1.41 | 1 |
|
1.5085 | 42.7 | 8.19 | 2 |
|
-0.5618 | 27.22 | 11.42 | 1 |
Environmental variables at local scale. Kinds of substrates found at each cave at
CAVE | Gruta do Grilão | Gruta do Lixo | Gruta do Sumidouro | Gruta Feneme | Gruta Helictites | Lagoa Seca | Lapa da Varzea | Lapa das Pacas | Macacos das Cavernas | Ninho de pérolas |
---|---|---|---|---|---|---|---|---|---|---|
|
0 | 0 | 2 | 0 | 0 | 0 | 3.5 | 0 | 0 | 0 |
|
0 | 0 | 1 | 0 | 0 | 0 | 0 | 28 | 0 | 0 |
|
0 | 3 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
|
3.5 | 3 | 10.5 | 11.5 | 0 | 5.5 | 12.5 | 0 | 2 | 1.5 |
|
17 | 5 | 3 | 6 | 17 | 37 | 0 | 13 | 7 | 0 |
|
1 | 5 | 2 | 0 | 2 | 2 | 0 | 0 | 8 | 3 |
|
0 | 9 | 9 | 12 | 2 | 2 | 0 | 2 | 16 | 6 |
|
8 | 3 | 2 | 5.5 | 2 | 0 | 0 | 3 | 12 | 9 |
|
3 | 2 | 2 | 8 | 3 | 2 | 0 | 2 | 12 | 12 |
|
15 | 12 | 7.5 | 5 | 18 | 4.5 | 2 | 1 | 15 | 1 |
|
17 | 4 | 5 | 2 | 4 | 6 | 3 | 0 | 8.5 | 0.5 |
|
0.5 | 0 | 5.5 | 0 | 6 | 0 | 44 | 68 | 3 | 58 |
|
0 | 9 | 10.5 | 15 | 1 | 11 | 25 | 8 | 11.5 | 4 |
|
8 | 10 | 12 | 8 | 9 | 8 | 6 | 8 | 10 | 9 |
|
0.2105 | 0.1332 | 0.1219 | 0.1563 | 0.2271 | 0.3242 | 0.3386 | 0.3622 | 0.1218 | 0.4048 |
|
1.709 | 2.148 | 2.295 | 1.963 | 1.754 | 1.527 | 1.307 | 1.338 | 2.189 | 1.347 |
At landscape scale we found four main types of land cover: limestone outcrop, cattle pasture, native vegetation and others. At all buffers the two principal land covers were pasture and native vegetation. The only buffer with water was the 250 m buffer of the Gruta do Sumidouro Cave (Table
Environmental variables at landscape scale. Percentage of each land-use type at all Buffer sizes.
Limestone Outcrop | Others | Cattle Pasture | Native Vegetation | Water | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
BUFFER AREA SIZE | 50 | 100 | 250 | 50 | 100 | 250 | 50 | 100 | 250 | 50 | 100 | 250 | 250 |
|
0.0 | 0.0 | 0.0 | 0.0 | 2.2 | 5.8 | 9.1 | 7.1 | 7.0 | 90.9 | 90.6 | 87.2 | 0.0 |
|
0.0 | 0.0 | 0.0 | 17.8 | 22.0 | 11.9 | 76.0 | 56.2 | 57.7 | 6.2 | 21.8 | 30.4 | 0.0 |
|
37.6 | 29.1 | 2.6 | 10.1 | 8.8 | 1.5 | 16.9 | 16.2 | 3.5 | 35.4 | 45.9 | 15.8 | 76.7 |
|
0.0 | 0.0 | 0.0 | 3.5 | 3.2 | 2.4 | 47.7 | 28.6 | 30.8 | 48.8 | 68.2 | 66.8 | 0.0 |
|
33.0 | 35.0 | 20.7 | 14.9 | 20.1 | 18.3 | 1.9 | 5.3 | 20.4 | 50.2 | 39.6 | 40.6 | 0.0 |
|
2.8 | 12.9 | 9.4 | 35.9 | 19.2 | 9.9 | 19.2 | 20.2 | 18.3 | 42.2 | 47.7 | 62.4 | 0.0 |
|
0.0 | 0.0 | 0.0 | 22.1 | 16.5 | 13.4 | 59.5 | 69.7 | 63.4 | 18.4 | 13.7 | 23.2 | 0.0 |
|
0.0 | 0.0 | 0.0 | 4.9 | 14.7 | 12.1 | 43.8 | 47.1 | 70.7 | 51.3 | 38.2 | 17.2 | 0.0 |
|
0.0 | 1.9 | 9.1 | 3.0 | 2.9 | 6.9 | 52.0 | 53.7 | 35.0 | 45.0 | 41.5 | 49.1 | 0.0 |
|
0.0 | 0.0 | 0.0 | 0.1 | 9.0 | 8.5 | 1.3 | 6.1 | 26.8 | 98.6 | 84.9 | 64.8 | 0.0 |
Limestone outcrop was the most important predictor variable of community composition (
Predictor Variables | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
Buffer 50 m | Buffer 100 m | Buffer 250 m | |||||||||
Selections | Selections | Selections | |||||||||
0.40 | Limest. Out., Others. | 0.45 | Limest. Out., Cat. Past., Nat. Vegetation. | 0.73 | Limest. Out., Water, |
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|
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p-value | Proportion | p-value | Proportion | p-value | Proportion | ||||||
|
0.008 | 0.3381 |
|
0.039 | 0.42614 |
|
0.012 | 0.53295 | |||
|
0.114 | 0.2406 |
|
0.84 | 1.9433E-2 |
|
0.935 | 8.3714E-3 | |||
|
0.996 | -4.5387E-2 |
|
0.792 | 2.3529E-2 |
|
0.263 | 0.13365 | |||
|
0.51 | 7.9518E-2 |
|
0.261 | 0.14627 |
|
0.895 | 1.3282E-2 | |||
|
0.413 | 0.12744 |
|
0.226 | 0.18016 |
|
0.203 | 0.35578 | |||
|
0.843 | -4.9594E-3 |
|
0.962 | 7.2133E-3 |
|
0.186 | 0.20154 | |||
|
0.595 | 6.742E-2 |
|
0.134 | 0.23179 |
|
0.936 | 6.5205E-3 | |||
|
0.099 | 0.29352 |
There are few studies on spatial patterns of cave communities although some studies evaluate differences of spatial scale sampling on species patterns (e.g.
Karst areas have different historical land uses and human impacts vary according to landscape characteristics (
The second factor that explained the cave similarity in the 50 m buffer was “
Detailed figure of Gruta Helictites showing different land uses within the 50, 100 and 250 m buffers.
The 100 m buffer indicated, in addition to limestone outcrops, cattle pasture and native vegetation as important variables. The landscape cover determines food availability inside caves, which possibly affects cave communities. Although one would expect to have richer communities in caves surrounded by forests (as a rich source of organic matter that can be brought inside caves), guano also constitutes an important resource for many cave invertebrates. Some species are highly dependent on guano, and cave communities associated to this resource can be relatively complex (
Considering the 250 m buffers, the model incorporated three different explanatory variables aside from the limestone outcrops: water bodies, environmental stability index and habitat heterogeneity. The importance of the allochthonous nutrient input through water transport is well known (e.g.
The higher community similarity among caves with similar values of
The habitat diversity hypothesis proposes that species diversity in a landscape will increase as the greater structural complexity increases, because of the higher resource abundance and the potential addition to the number of partitionable niche dimensions (
Although the landscape scale explains better species composition, the local scale model suggests an influence of the habitat heterogeneity and stability on cave community. The variables
Landscape use can even make terrestrial troglophile populations more isolated, severely reducing their dispersal possibilities, by conversion from forest to pasture (
Considering the current Brazilian legislation (Brazil. Decree no 6.640/2008), there is an obligation to protect the area corresponding to the cave linear projection on the surface and also a radius of 250 meters around this projection (Portaria IBAMA no 887/1990). Unfortunately, there are no studies showing an eventual efficiency of such radius to preserve cave communities. Our study could be the first step to improve Brazilian legislation, since it provides a new methodology to evaluate three aspects: cave invertebrate communities, cave physical traits and surface land use.
We sincerely thank Rogério Tavares, who is manager of “Parque Estadual do Sumidouro” and the Instituto Estadual de Florestas de Minas Gerais (IEF) for logistic support. We also thank Dr. Marcelo Passamani, Msc. Marcus Paulo de Oliveira, Msc. Maricélio Medeiros, Msc. Victor Hugo Oliveira for support and valuable suggestions in the field and specially Dra. Carla Ribas, Dr. Paulo Pompeu, Dr. Júlio Louzada and Rafaela Bastos Pereira for assistance in writing this paper. This research paper was partly produced during the discipline PEC 527 – Scientific Publication and also PEC 813/814 - Field Curse, from the Post-Graduate School in Applied Ecology, at Federal University of Lavras. The authors were financed by CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior). Funding was provided by the Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) and Conselho Nacional de Pesquisa to R.L.F. (CNPq grantnr. 304682/2014–4). Finally we would like to thank Dr. Claudio Di Russo, Dr. Leonardo Latella and Dra. Oana Moldovan, for their suggestions, which certainly improved the work. Authors declare that all experiments comply with the current Brazilian laws, in which the work was performed.