**2. Methods**

in some places due the enhanced agricultural practices [2]. As a result of this growing phenomenon, the previously natural landscapes have been turned into mosaics of natural and seminatural lands embedded into human-modified landscapes [3]. Actually, these mosaic conditions of non-used land represent, almost 90% of the world's tropical forests which are inserted in reserves and parks within agricultural lands [4, 5]. Understanding the factors that affect the community assemblies has been the focus of many ecological studies [6–9]. Since the conversion of the previously undisturbed places is usually allied to unsustainable activities, which drives the ecosystem degradation throughout the loss of ecosystem services and the related cascade events [10] causing biodiversity loss across the globe and across spatial,

To understand these factors, some researchers have made use of a classical measure, the taxonomic unit (e.g., species) [10, 14], but it does not take into account ecological and evolutionary attributes of species. However, some modern approaches are combining functional attributes of species (individual characteristics that can be measured and that affect the fitness) with phylogenetic relationships. This approach, in addition to the taxonomic diversity, can bring different answers of a species community in relation to its habitat conditions, being these a combination of ecological and evolutionary answers [15, 16]. Environmental degradation process can be observed by studying diversity measures that are affected by disturbance conditions [17], and for this, the usage of functional, phylogenetic and taxonomic diversity is a growing tool that has been changing the focus of researchers from the use of species diversity or species composition that take no account of differences in species' life-history traits and ecological niches [17–19].

Although plenty of studies have shown strong relationships between community structure and environmental predictors and how the functional traits of species can match up with the environmental conditions [20], some adaptive processes remain unclear. This may be due the large number of traits presented by each species and/or the high species number existing in many habitats which generates an incomplete knowledge of which species traits can actually be an influence to the ecosystem processes [21]. Among all vertebrates, the amphibians are the group with highest proportion of species threatened with extinction [22], due to habitat loss, fragmentation [23], and other related environmental stressors like enhanced UV radiation incidence [24, 25] and canopy coverage loss [26]. Furthermore, the complexity on amphibian life cycle and the differences in life-history strategies between species and also their habitat associations generate a need for studies aiming to understand the true relationship between anthropogenic disturbance and the structure and organization of amphibian communities [7, 8].

In the present study, we aim to answer the following question: In relation to the anthropogenic disturbance in an agricultural-forest preserved gradient, would ponds in more preserved environment harbor higher taxonomic, functional and phylogenetic diversity patterns? So we tested the hypothesis that ponds located at most preserved and more heterogeneous environments would be taxonomically richest and would allow the coexistence of more functionally distinct species [27], expecting then a higher functional diversity and lower functional redundancy. We also expect an increase of the phylogenetic diversity (and thus a decrease in phylogenetic redundancy) at these sites, since more heterogeneous habitats can provide a wide range of microhabitat usage, diminishing the interspecific competition and allowing the

coexistence of taxa with higher phylogenetic similarity [28, 29].

temporal and organizational scales [11–13].

22 Tropical Forests - New Edition

#### **2.1. Description of the study site**

The sampling areas are located at the Parque Estadual do Turvo (PET) and its adjacencies, and both belong to the Atlantic Rainforest biome. The PET is located at the Rio Grande do Sul State (27° 07′–27° 16′ S, 53° 48′–54° 04′ W; 100–400 a.s.l), at the municipality of Derrubadas, covering an area of 17,491 ha with about 90 km of perimeter of semi-deciduous forest. The study site differs from the wet evergreen forests since it is dryer and presents more open areas, sharing this same vegetational classification with the Republic of Argentina by the Moconá Provincial Park (about 1000 ha) and the Yabotí International Biosphere Reserve (236,613 ha), as well the Brazilian state of Santa Catarina by the Uruguay River [30] (**Figure 1**).

The vast majority of the surrounding areas of PET were converted into intensively agricultural landscapes dominated by crops of soybeans (~22.000 t/year), maize (7.560 t/year), wheat (6.840 t/year) and cattle (~8700 animals) pasture and where the legal buffer zones are not implemented or respected [31]. The climate is characterized as subtropical highly humid with average rainfall between 1.700 and 1.900 mm with reduction of precipitation at the winter season, and the average of temperature ranges from 20 to 23°C [32].

#### **2.2. Data collection**

The fieldwork was conducted during two anuran breeding seasons at southern Brazil, the first one from September 2013 to March 2014 and the second from September 2015 to March 2016, which comprises the spring and summer seasons at the southern hemisphere. The field

**Figure 1.** Map representing the sampling area on the extreme north-western of the Rio Grande do Sul State, Brazil. The white points on the map show location of the sampled ponds along the agricultural-preserved forest gradient at Parque Estadual do Turvo and surrounds. The study was performed at two consecutive breeding seasons of anurans between 2014 and 2016.

campaigns were made monthly, for approximately 10 days, when we sampled 38 ponds following an environmental gradient. The gradient ranged from the agricultural landscape where 19 ponds were located (outside of the park borders) to a preserved undisturbed forest, the inner portion of the gradient, with 19 ponds too (**Figure 1**). We collected adult anurans by using the method of "survey at breeding sites" [33], recording the number of calling males along all perimeter of ponds. Then, the maximum abundance data from each species in each pond was used to construct the composition matrix to be used at the subsequent analysis.

species (*Elachistocleis bicolor*, *Ololygon aromothyella*, *Physalaemus* aff. *gracilis*, *Rhinella ornata*, *Scinax granulatus* and *S. perereca*) not present at Pyron and Wiens' work [34]. The position of these insertions (missing species) was defined according to the position of the closest species

Enhanced Phylogenetic Diversity of Anuran Communities: A Result of Species Loss in…

http://dx.doi.org/10.5772/intechopen.72256

25

We calculated the functional and phylogenetic patterns by using Rao's quadratic entropy and the taxonomic diversity by using the Gini-Simpson's index [39, 40]. These analyses were based on [15, 16] by constructing a dataset composed by four matrices. The first one (matrix B) contains the species functional traits, the second one (matrix W) contains the abundance of species in each sampled pond, the third one (matrix E) with the environmental filter (distance from the nearest border of PET, negative values for outside and positive values for inside) and the fourth (matrix F) with the phylogenetic information (transformed then into a matrix of phylogenetic distance) of the recorded species. To perform these analyses, we used the software Phylocom [41] and SYNCSA (available at http://ecoqua.ecologia.ufrgs.br/SYNCSA.html).

As a way to explore our database and better understand the effects of richness and equability of the species distributed along the measured gradient of distance, we constructed a Whittaker diagram (or dominance diagram). After these procedures, we tested the relation of the obtained values of functional diversity and redundancy, taxonomic diversity and redundancy and phylogenetic diversity and redundancy of each pond with its distance from PET's nearest border (positive values represented the ponds inside PET's area and negative values represented ponds outside PET's boundaries). In addition, we also tested the relation of the components of the community weighted means matrix (CWM matrix containing the weighted functional traits) with the distance from the PET's nearest borders (*Vegan* Package,

We found 15 anuran species from five families: Hylidae (four species), Leptodactylidae (four species), Bufonidae (two species), Phyllomedusidae, Microhylidae and Ranidae, both with one species each. We registered all the 15 species in the inner portion of gradient (the portion inside the PET) and only 10 species in the outside portion. The most conspicuous species were *Dendropsophus minutus* and *Scinax granulatus* both occurring at 31 of the 38 sampled ponds, respectively (**Table 2**). We found, based on the abundance distribution curve, that the ponds located at inner portion of the gradient have the species abundance more equally distributed

Regarding the taxonomic, functional and phylogenetic patterns of diversity that we analysed, only the phylogenetic diversity and phylogenetic redundancy were related to the studied gradient (r2 = 0.14, p > 0.05 and r2 = 0.20, p < 0.05, respectively). The phylogenetic diversity (opposed to what we assumed) decreased at the inner portion of the gradient, while the phy-

(equability) than the ponds located at the outside portion (**Figure 2**).

logenetic redundancy increased (see **Figure 3A** and **B**).

or closest species group [35–38].

**2.3. Statistical analysis**

*lm* function, [42]).

**3. Results**

We undertook a series of 14 measures (averages from the continuous values, chosen given their environmental and/or reproductive values) on morphological and ecological traits from eight individuals of each recorded species, to access data on functional diversity and redundancy (**Table 1**). Since the sampling method is based on the calling males, the morphometric measures were taken only from adult males. The data acquisition was performed from anurans collected during the field campaigns and also on specimens already deposited at the Universidade Federal de Santa Maria collection (ZUFSM Appendix A) between the years of 2010 and 2012 from the same area, to enlarge the database.


We also constructed a phylogenetic matrix based on the phylogenetic information of the species to access data on phylogenetic diversity and redundancy. We manually inserted six

**Table 1.** Description of the ecomorphological traits of the anurans recorded in ponds monitored at Parque Estadual do Turvo and adjacencies between 2014 and 2016.

species (*Elachistocleis bicolor*, *Ololygon aromothyella*, *Physalaemus* aff. *gracilis*, *Rhinella ornata*, *Scinax granulatus* and *S. perereca*) not present at Pyron and Wiens' work [34]. The position of these insertions (missing species) was defined according to the position of the closest species or closest species group [35–38].

#### **2.3. Statistical analysis**

campaigns were made monthly, for approximately 10 days, when we sampled 38 ponds following an environmental gradient. The gradient ranged from the agricultural landscape where 19 ponds were located (outside of the park borders) to a preserved undisturbed forest, the inner portion of the gradient, with 19 ponds too (**Figure 1**). We collected adult anurans by using the method of "survey at breeding sites" [33], recording the number of calling males along all perimeter of ponds. Then, the maximum abundance data from each species in each pond was used to construct the composition matrix to be used at the subsequent analysis.

We undertook a series of 14 measures (averages from the continuous values, chosen given their environmental and/or reproductive values) on morphological and ecological traits from eight individuals of each recorded species, to access data on functional diversity and redundancy (**Table 1**). Since the sampling method is based on the calling males, the morphometric measures were taken only from adult males. The data acquisition was performed from anurans collected during the field campaigns and also on specimens already deposited at the Universidade Federal de Santa Maria collection (ZUFSM Appendix A) between the years of

We also constructed a phylogenetic matrix based on the phylogenetic information of the species to access data on phylogenetic diversity and redundancy. We manually inserted six

Snout vent length Continuous Total size of the body, from the tip of the nose to the cloaca Mouth ratio Continuous Distance between the rear edge of the jaw joint and the tip

Forelimb ratio Continuous Greater distance from the "shoulder" to the tip of the

Hind limb ratio Continuous Distance between the cloaca and the tip of the "foot,"

**Table 1.** Description of the ecomorphological traits of the anurans recorded in ponds monitored at Parque Estadual do

Perched Binary Place of activity/or vocalization Ground Binary Place of activity/or vocalization Water Binary Place of activity/or vocalization Prolonged breeding Binary Time of breeding season Explosive breeding Binary Time of breeding season Reproductive Mode 1\* Binary Reproductive mode Reproductive Mode 2\* Binary Reproductive mode Reproductive Mode 11\* Binary Reproductive mode Reproductive Mode 24\* Binary Reproductive mode Reproductive Mode 30\* Binary Reproductive mode

of the snout, divided by the snout vent length

"hand," divided by the snout vent length

divided by the snout vent length

2010 and 2012 from the same area, to enlarge the database.

**Trait type Variable Trait**

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\*

Reproductive modes based on [43].

Turvo and adjacencies between 2014 and 2016.

We calculated the functional and phylogenetic patterns by using Rao's quadratic entropy and the taxonomic diversity by using the Gini-Simpson's index [39, 40]. These analyses were based on [15, 16] by constructing a dataset composed by four matrices. The first one (matrix B) contains the species functional traits, the second one (matrix W) contains the abundance of species in each sampled pond, the third one (matrix E) with the environmental filter (distance from the nearest border of PET, negative values for outside and positive values for inside) and the fourth (matrix F) with the phylogenetic information (transformed then into a matrix of phylogenetic distance) of the recorded species. To perform these analyses, we used the software Phylocom [41] and SYNCSA (available at http://ecoqua.ecologia.ufrgs.br/SYNCSA.html).

As a way to explore our database and better understand the effects of richness and equability of the species distributed along the measured gradient of distance, we constructed a Whittaker diagram (or dominance diagram). After these procedures, we tested the relation of the obtained values of functional diversity and redundancy, taxonomic diversity and redundancy and phylogenetic diversity and redundancy of each pond with its distance from PET's nearest border (positive values represented the ponds inside PET's area and negative values represented ponds outside PET's boundaries). In addition, we also tested the relation of the components of the community weighted means matrix (CWM matrix containing the weighted functional traits) with the distance from the PET's nearest borders (*Vegan* Package, *lm* function, [42]).
