Dung beetles ( Coleoptera , Scarabaeinae ) from high-altitude grasslands in São Joaquim National Park , Santa Catarina , southern Brazil

São Joaquim National Park (SJNP), in southern Brazil, covers large areas of high-altitude grasslands (HAG), which are a conspicuous ecosystem that belongs to the Atlantic Forest domain. Previous studies recorded 6 species of dung beetles (Coleoptera, Scarabaeinae) in SJNP. Dung beetles were sampled using a standardized protocol (baited pitfall traps) at 6 HAG sites in January 2016. In this paper, an annotated list of the dung beetle species sampled in SJNP is presented. This list includes previous literature records and ecological and distributional information of the species. Nine species of Scarabaeinae were collected, including 8 of them newly recorded from SJNP. The dung beetle assemblages were composed of species having diurnal habits and colorful bodies, and showing a similar proportion of roller–tunneller, and coprophagous–trophic generalist species. New studies could substantially increase the number of dung beetle species in the SJNP.


Introduction
The Atlantic Forest is one of the richest Brazilian biomes.However, it also is one of the most threatened regarding conservation of its biodiversity (Myers et al. 2000, Tabarelli et al. 2010).The high levels of biodiversity and endemic species found in the Atlantic Forest have been attributed to the high environmental and climatic heterogeneity found in this biome (Ribeiro et al. 2009, Tabarelli et al. 2010), which comprises areas from Rio Grande do Norte to Rio Grande do Sul along the east coast of Brazil (ca 29° range in latitude), and also northern Argentina and southern Paraguay (Tabarelli et al. 2010).
The grasslands found at high altitudes in southern Brazil (campos de altitude or high-altitude grasslands) are a very conspicuous environment in the Atlantic Forest domain.High-altitude grasslands (HAG) are typical of montane and high montane areas with shrub and/or herbaceous structure, and usually occur on lithic mountaintops at high elevations, predominantly in subtropical ANNOTATED LIST OF SPECIES and temperate climate (Brasil 1993).In southern Brazil, these grasslands are distributed from northern Rio Grande do Sul to Paraná states and form mosaics with forest formations.Despite increased research on the composition and ecology of these grasslands in recent decades, few studies have focused on the importance of biodiversity and conservation (Iganci et al. 2011, Barros et al. 2015, Overbeck et al. 2015).
In this region, fires occur naturally and the use of fire to clear grasslands in late winter was a traditional practice.These grasslands have tussock grasses that are more tolerant to fire, such as species of Andropogon L., Schizachyrium Ness, Aristida L., and Baccharis L. (Overbeck et al. 2015).Another important component in the evolution of old-growth grasslands is herbivory, especially cattle grazing (Veldman et al. 2015).Horses and cattle were introduced in the region by Jesuit missionaries in the 17th century (Pillar and Quadros 1997), and livestock became an important land use in southern Brazil and remains so today (Overbeck et al. 2007).However, species of deer, rhea, and rodents are natural herbivores in these environments, and there also is fossil evidence that large herbivorous Equidae, Camelidae, and Cervidae existed in the region until 8000 years before present (Pillar and Quadros 1997, and references therein).Due to the high altitudes and their geographic position at around 30° S latitude in southern South America, HAG faced severe climate regimes during their evolution (Overbeck et al. 2007).Nowadays, this region is in a tropical-temperate transition zone, with hot summers, cool winters, and no dry season (Overbeck et al. 2007).Thus, the biodiversity found in HAG is due to historical and biogeographical processes, and current land use adopted by landowners for centuries.Features such as fire, grazing, and soil characteristics limit tree growth in old-growth grasslands (Rehm andFeeley 2015, Veldman et al. 2015).
In southern Brazil, São Joaquim National Park (SJNP) is a protected area on the South Brazilian Plateau that covers large areas of HAG and other vegetation formations, such as nebular forest, dense and mixed ombrophilous forest (also called Araucaria forest) (Brasil 1961, Castilho et al. 2014, ICMBio 2016).The altitude varies between 350 and 1822 m above sea level and several grassland areas within the park that have not been expropriated continue to be grazed by livestock (ICMBio 2016).In southern Brazil, HAG show high levels of plant species richness and endemic plant species (ca 25% of their taxa) (Boldrini et al. 2009, Iganci et al. 2011), which are important for conservation and for studies on patterns of diversification in a subtropical transitional environment (Iganci et al. 2011, Barros et al. 2015).Despite being important for conservation, the park lacks a management plan and continuous biodiversity surveys (Vaz-de-Mello et al. 2014b).
The first inventory of scarabaeoidean beetles in the park was made by Vaz- de-Mello et al. (2014b), who found 14 species belonging to 4 families: Lucanidae (5 species), Melolonthidae (2 species), Scarabaeidae (6 spe-cies), and Trogidae (1 species).The low species richness of Scarabaeoidea was attributed to the low temperatures, and the data were considered insufficient for a robust assessment of the fauna of this group in the park (Vazde-Mello et al. 2014b).Thus, there is a need to better know the scarabaeoidean fauna (and coleopteran fauna as a whole) occurring in the park, with the aim of providing new information on biodiversity and the distribution of species.
Regarding the family Scarabaeidae, all species sampled by Vaz- de-Mello et al. (2014b) belonged to the subfamily Scarabaeinae, a taxonomic group that is popularly known as dung beetles.The term "dung beetle" is generally used to describe species of Scarabaeoidea with essentially coprophagous habits, such as Scarabaeinae, Aphodiinae, and Geotrupidae (considered Geotrupinae by some authors) (Lobo and Halffter 2000).In this study, "dung beetle" is used to refer only to the subfamily Scarabaeinae, which is the predominant group sampled using baited pitfall traps (Halffter and Favila 1993).
Dung beetles feed primarily on mammal feces, animal carcasses, and rotting plant matter.They have an important role in the decomposition of organic and nutrient cycling (Halffter and Matthews 1966, Halffter and Edmonds 1982, Hanski and Cambefort 1991).These beetles bury portions of food resources in the soil, making the nutrients in these materials available to the ecosystem once again, and also increasing aeration and water infiltration; they also can bury eggs of other organisms (such as parasites of cattle) and secondarily disperse fruit seeds consumed by mammals (Nichols et al. 2008, and references therein).
Vaz- de-Mello (2000) compiled a list of the Scarabaeinae in Brazil and found 618 species cited in publications, but this number is greatly an underestimate due to the lack of surveys in several regions of Brazil and the need for taxonomic revisions of some groups.Nowadays, there are 726 valid species occurring in Brazil, with 62 endemic species (Vaz-de-Mello 2017).The Scarabaeinae present species with a wide range of sizes, colors, and body shapes.There are genera with species just a few millimeters long, such as in Besourenga Vaz- de-Mello, 2008, Canthonella Chapin, 1930, and Degallieridium Vaz-de-Mello, 2008(Ratcliffe and Smith 1999, Vazde-Mello 2008) and species with about 5 cm, such as some species belonging to the genus Coprophanaeus Olsoufieff, 1924 (Edmonds andZídek 2010).The body color in Scarabaeinae varies from opaque black to a range of metallic colors (Halffter and Matthews 1966), which can help prevent predation, allow intraspecific communication, or be related to activity period (Young 1984, Vulinec 1997, Hernández 2002).The body shape also varies, from oval (most species) to rectangular (such as Eurysternus Dalman, 1824 and Dendropaemon Perty, 1830) and represent body adaptations related to the foodhandling and nesting behavior and to reduce competition (Hernández et al. 2011).The diversity and combinations of body traits found in Scarabaeinae beetles contribute to Methods Study area.SJNP (Fig. 1) is a 49,300 ha protected area located on the South Brazilian Plateau in Santa Catarina state; the region is called the Serra Geral.The park is contained within 4 municipalities: Bom Jardim da Serra, Grão Pará, Orleans, and Urubici (Brasil 1961, ICMBio 2016).SJNP, which is located in the Atlantic Forest biome, covers extensive areas of high-altitude grasslands (in Portuguese, Campos de Altitude), Araucaria forest (Araucaria angustifolia (Bertol.)Kuntze), and also nebular forest and dense ombrophilous forest.The park as a range of altitude from 350 to 1822 m above sea level (ICMBio 2016).The highest point in the park is Morro da Igreja (MI) in the park's northeast region.At the center of the park, there are also areas above 1650 m that are named Campos de Santa Bárbara (SB).The climate in the high grasslands is humid subtropical, with hot summers, cool winters (with frequent frosts and snow), and without a pronounced dry season (Overbeck et al. 2007(Overbeck et al. , 2015)).Summers are cooler in the South Brazilian Plateau than summers at low altitudes, with the annual average temperature around 16-22 °C.The annual averages of precipitation and air relative humidity are 1400 mm and 85%, respectively.The geological formation of the park the high diversity of the group.
Dung beetles have been used as biological indicators because they are sensitive to natural or anthropogenic environmental changes (Halffter andFavila 1993, Nichols et al. 2007).Several studies have shown that assemblages from open and forest environments may be quite different (Klein 1989, Costa et al. 2013), and these differences may be greater or lesser depending on the predominant type of matrix.Open environments are expected to be occupied by specialized species that require more sunlight, a warmer temperature, and less humidity (Gossner 2009).However, species of the HAG face climatic conditions quite different from those living in open habitats near or at sea level.Due to historical and biogeographical processes, communities in HAG evolved to support extreme environmental and climatic conditions.
The aim of this study was to know the species composition of scarabaeine assemblages from HAG in the SJNP by using a standardized sampling protocol.I also aim to characterize the assemblages and their species using body traits and ecological characters.In the HAG of southern Brazilian, one would expect that scarabaeine assemblages would be composed of open-habitat specialists, with behavior, daily activity, and trophic preference adapted to survive in environments with extreme climatic conditions. is composed of volcanic rocks (basalt), which together with sandstone formations, is permits the discharge and recharge of the Guarani Aquifer.Studies date the age of these formations as approximately 133 million years (ICMBio 2016).
The 6 HAG sites sampled are open environments, formed by herbaceous and/or shrubby vegetation structure, which is mainly represented by species of Poaceae, Asteraceae, and Cyperaceae (Pillar andQuadros 1997, Overbeck et al. 2007).These are humid or semi-humid grasslands (Zanin et al. 2009, Overbeck et al. 2015); the soil of all sites was always wet during the sampling period.This is due to the constant influence of fog and/ or the high concentration of rainwater in shallow soils (Overbeck et al. 2015), which occur at top of mountains and plateaus.
Dung beetle sampling.Dung beetles were sampled in 6 HAG sites, 3 of them in MI and another 3 in SB (Table 1, Fig. 1).These 2 areas are 12 km from each other.In each area, individual sampling sites were on average 900 m from one other (range 300-1300 m).To sample dung beetles, baited pitfall traps, which consisted of 750 ml plastic containers (15 cm diameter, 20 cm depth) buried with their edge level with the ground.Above the traps, a rain guard was placed to prevent trap overflow and to support the bait.In each trap, a solution of water and neutral detergent (300 ml) was added to catch beetles.Human feces and rotting flesh (20 g) were used as bait to attract coprophagous and necrophagous species, respectively.Baits were wrapped in thin cloth and tied in the central portion of the rain guard.
In each site, 10 baited pitfall traps were distributed in pairs, with each pair spaced 100 m apart (da Silva and Hernández 2015b).Paired traps were spaced 5-10 m apart, and each one was baited with human feces or rotten flesh.Each pair of traps was considered a sampling unit.All traps remained in the field for 48 h.Similar sampling designs were previously in dung beetle studies in southern Brazil (da Silva and Hernández 2014, 2015a, 2016).
The study was carried out between 11-14 January 2016.In the first day, 10 baited pitfall traps were mounted in each of the 3 sites from SB.In the second day, the same for the 3 sites from MI.In the last 2 days, after 48 h sampling period for each site, dung beetles were collected.The accumulated rainfall was 14 mm (6 mm and 8 mm for the first and second days).The relative humidity ranged 60-82% during the sampling period.The minimum temperature varied between 14 °C and 16 °C, while maximum temperature was 25-27 °C (Climatempo 2016).Dung beetles were sampled in January (summer in the southern hemisphere) because this month has high temperatures and a high species richness and abundance of Scarabaeinae in southern Brazil (Hernández andVazde-Mello 2009, da Silva et al. 2013).
All collected beetles were sorted, mounted on entomological pins, and dried in an oven (60 °C for 72 h).They were identified using dichotomous keys and species diagnoses (Harold 1867, 1868, 1869b, Schmidt 1922, Luederwaldt 1929, Boucomont 1932, Balthasar 1939, Pessôa and Lane 1941, Edmonds 2000, Edmonds and Zídek 2010, Vaz-de-Mello et al. 2011, Maldaner et al. 2015).A leading specialist in scarabaeine taxonomy, Dr Fernando Vaz de Mello (Universidade Federal de Mato Grosso, Cuiabá, Brazil) confirmed all identifications.In recent years, Dr Vaz de Mello had performed a broad study of the types of species of Neotropical Scarabaeinae deposited in European museums and, therefore, can assure the identity of the species in this study.The study did not involve protected species and all permits to collect dung beetles were granted prior to the study by the Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio, permit #49486-1).The voucher specimens are deposited in the Entomological Collection of the Centro de Ciências Biológicas in the Universidade Federal de Santa Catarina and in the Entomology Section of the Zoological Collection of the Universidade Federal de Mato Grosso, Cuiabá, Brazil.Voucher specimens sampled in this study were labeled with a number unique to each individual specimen.Data analysis.Dung beetles were grouped in categories of food relocation behavior, trophic preference, and body color following the specialized literature (Halffter and Matthews 1966, Halffter and Edmonds 1982, Cambefort 1991, Hanski and Cambefort 1991, Hernández 2002, Campos et al. 2011, Lopes et al. 2011, Medina and Lopes 2014, da Silva and Hernández 2015a) and field observations.Species accumulation curves were used to verify the sampling sufficiency (Colwell et al. 2004) between the 2 sampling areas (MI and SB) and for the entire dataset, representing the assemblage of dung beetles found in SJNP.The curves were calculated using the 'specaccum' function and the method 'exact' of the Vegan package (Oksanen et al. 2016) in the program R 3.1.2(R Core Team 2017).The Chao 1 species richness estimator (and its confidence interval), which is based on species abundance, was calculated using EstimateS 9.1 program (Colwell 2013).The Chi-square test was used to test for differences between the proportions of rollers and tunnellers found in the SJNP.Citations to the literature are not exhaustive because this work is neither a taxonomic study nor catalog of species.Only the main references to published synonymies and recent ecological and taxonomic data are included.
The number of species was similar between the types of baited pitfall traps: 6 species found in traps baited with human feces and 5 in traps with rotten flesh.However, the number of individuals found in traps with human feces was almost double than that found in traps baited with rotten flesh.Among localities, the highest number of species (8) was found in SB (MI = 5 species).However, MI showed a greater number of individuals (N = 105) than SB (N = 47).Four species were sampled only in SB, with all being singletons.
None of the species accumulation curves, for each locality and for the whole park, tended to stability, demonstrating a high probability of encountering other dung beetle species in the SJNP (Fig. 3).The Chao 1 estimator also corroborated these results, especially for SB and the whole Park, where only 44.5% and 60% of the estimated numbers of species were sampled, respectively (Fig. 3).

Discussion
The number of species found in HAG was very low when compared with studies performed in similar open formations in southern Brazil.Using baited pitfall traps several authors found between 16-30 species of dung beetles in natural grasslands in the Brazilian Pampa (da Silva et al. 2008aSilva et al. , 2009bSilva et al. , 2012a)).In the same region, even in a eucalyptus area in a matrix originally dominated by natural grasslands, Audino et al. (2011) found 28 species of dung beetles.However, the altitude in those areas was about 220 m above sea level.The low species richness of dung beetles is attributed to the severe climatic and associated environmental conditions found in SJNP, as previously stated by Vaz- de-Mello et al. (2014b).Based on sampling sufficiency curves, there is a great potential to find other species in the park, especially using longer sampling periods and/or other sampling methods.
From those species sampled in our study, only Coprophanaeus (Metallophanaeus) saphirinus and Di cho tomius (Luederwaldtinia) opalescens were not previously recorded in the southern Brazilian Pampa (see the discussion on species distribution below).The high similarity found between open grasslands in Brazilian Pampa and HAG in southern Brazil is due to the biogeographic history of these formations.Between 42,000 and 10,000 years before the present, cold and dry weather prevailed in the region, and the grassland formations dominated the region (Boldrini 2009).The forests were restricted to small patches in bottom valleys.Between 10,000 and 4000 years ago, temperatures increased, but the climate remained dry, thereby limiting the expansion of forest areas on the grasslands (Boldrini 2009).In addition, at the beginning of the Holocene (12,000-10,000 years ago), there is evidence of more frequent fires, which also slowed the advance of tree species (Behling et al. 2004).Four thousand years ago, when the climate became wetter, the Araucaria Forest began a gradual process of expansion over the grasslands, which became more significant by about 1000 years ago (Behling 2002, Behling et al. 2004).Thus, open formations, now belonging to the Pampas and HAG, were connected by a long period allowing the widespread distribution of dung beetle species (and of other organisms) inhabiting those vegetation formations, despite the difference of altitude since both regions shared severe climatic and environmental conditions.
Coprophanaeus d'Olsoufieff, 1924.This genus was revised by Edmonds and Zídek (2010) and comprises at least 43 valid species distributed among 3 subgenera (Coprophanaeus s. str. Olsoufieff, 1924, Metallophanaeus Olsoufieff, 1924, and Megaphanaeus Olsoufieff, 1924) in the New World (Cupello and Vaz-de-Mello 2014).The subgenus Metallophanaeus comprises 9 colorful species distributed from eastern Brazil (east of Amazonia) southward to northeastern Argentina (Edmonds and Zídek 2010).All species of this subgenus are diurnal and copronecrophagous.Coprophanaeus (Metallophanaeus) saphirinus is a very common species in forest habitats along the Paranaian subregion of the Neotropical region (Edmonds and Zídek 2010).However, it is also found in open areas such as Chaco formations in northeastern Argentina (Edmonds and Zídek 2010).The only individual sampled in SJNP attracted to rotten meat is a red individual of C. (M.) saphirinus.In the southern portion of the species range, the red individuals occur more frequently in higher elevations than does blue or green ones, but they are also found at the sea level and in the northern portion of the species' range in sympatry with blue and green individuals (Cupello and Vaz-de-Mello 2014).Coprophanaeus (M.) saphirinus is attracted to herbivorous feces (Martínez 1959), humans feces, rotten meat, rotten fruit (Almeida and Louzada 2009, da Silva et al. 2011, 2012b, Lima et al. 2015), rotten fish (da  2016).That is the case of Dichotomius (Luederwaldtinia) opalescens (Felsche, 1910), the only species of this genus sampled in the SJNP so far.This species belongs to the D. (L.) speciosus (Waterhouse, 1891) species group, which comprises four species restricted to areas above 1,000 m altitude in the Brazilian Atlantic Forest (Maldaner et al. 2015).Dichotomius (L.) opalescens is restricted to the Serra Geral mountain range (the northern Rio Grande do Sul and southern Santa Catarina states), and thus it is considered as Endangered (EN), according to the conservation status based on the extent of occurrence applied by the International Union for Conservation of Nature (IUCN) (Maldaner et al. 2015).However, its distribution covers at least three national conservation units in southern Brazil: São Joaquim National Park, Aparados da Serra National Park, and São Francisco de Paula National Forest (Maldaner et al. 2015).The biology of this species is poorly known.All individuals were sampled in traps baited with human feces, suggesting a coprophagous habit.This species has also been sampled in Araucaria forest and eucalyptus plantations (Maldaner et al. 2015), suggesting a generalist environmental distribution in the Serra Geral mountain range.
Onthophagus Latreille, 1802.This genus is globally distributed and includes about 2000 species (Tarasov and Kabakov 2010), most considered coprophagous, with 160 species in the Americas (  (França et al. 2016).Individuals of this species were sampled only in pitfall traps baited with human feces in the SJNP.However, this species-complex is distributed for a wide range of environments in the South America, being attracted by several kinds of baits (França et al. 2016, and references therein).
In addition to the previously mentioned species, Vaz- de-Mello et al. (2014b) found Canthon (Canthon) angularis Harold, 1868, Canthon (Canthon) quadri punctatus Redtenbacher, 1868, an unidentified species of Canthon, Ateuchus hypocrita Balthasar, 1939, an unidentified species of Homocopris, and also D. (L.) opalescens (Felsche, 1910)  The assemblages of dung beetles found in high altitude grasslands in the São Joaquim National Park were composed of diurnal species with colorful bodies (both small and large), with similar proportions of rollers and tunnellers, and coprophagous and/or generalist trophic habits.Due to the severe climatic conditions found in the Park, longer sampling periods should be performed aiming to achieve a more robust sampling sufficiency for dung beetle species (da Silva and Hernández 2015b).The importance of using Coleoptera in the development of public environmental strategies such as management plans and creation of protected areas is highlighted (Vazde-Mello et al. 2014b) since this order is the most diverse group in number of animal species.

Table 1 .
Details of the sample sites in high-altitude grasslands of São Joaquim National Park, Santa Catarina, southern Brazil.
(Nunes and Vaz-de-Mello 2013;Maldaner et al. 2015;Nunes et al. 2016)a has 65 valid species assigned to 13 species-groups(Nunes and Vaz-de-Mello 2013;Maldaner et al. 2015;Nunes et al. 2016).Some Dichotomius species show extremely restricted distributions(Nunes and Vaz-de-Mello 2013;Maldaner et al. 2015; Nunes et al. Latreille, 1807 and  Palaeonthophagus Zunino, 1979)and an incertae sedis species, including species introduced from African and Europe (Vaz-de-Mello et al. 2011).Onthophagus (Onthophagus) aff.hirculus Mannerheim, 1829 belongs to a group of species taxonomically close to O. (O.) hirculus and needs urgent taxonomic revision Rossini and Vaz-de-Mello 2016).In the Americas, the genus comprises species of two subgenera (Onthophagus s. str. (Génier 2015) et al. 1998)iPereira, 1953).Those authors sampled Scarabaeoidea beetles using different methods in five sites, both in open and forest sites, in the MI, from 1,370 to 1,750 m a.s.l.Ateuchus Weber, 1801 has copro-necrophagous species inhabiting forest and grasslands in the Neotropical region(Vaz-de-Mello 1999).Some Ateuchus species are associated with ant nests(Vaz-de-Mello et al. 1998)or mammalian borrows(Génier 2015).Homocopris Burmeister, 1846 was recently reassigned as a valid genus, with few species under a reviewing process.Its species are generally found associated with altitude forests or open areas from Argentina,Chile, Colombia, Ecuador,  and southern Brazil (Vaz-de-Mello et al. 2010).In Brazil, Homocopris species are associated with well-preserved Atlantic Forest sites with high altitudes (in general, above 1000 m).