Range extension and first record of Euryzygomatomys spinosus ( Rodentia , Echimyidae ) in the Brazilian Cerrado

We present herein the first record of Euryzygomatomys spinosus (G. Fischer, 1814) in the Brazilian Cerrado, based on two adult male specimens collected in a “campo limpo” (dry grassland) area at Sempre Vivas National Park, state of Minas Gerais, southeastern Brazil, in June and August 2014. This taxon was previously known only from the Brazilian Pampas, Paraguayan Chaco Humedo and Atlantic Forest of southeastern and southern Brazil, northeastern Argentina and southern Paraguay. Our records increased the species geographic range 250 km northward, into the Cerrado ecoregion.


INTRODUCTION
Spiny rats from the family Echimyidae are represented by 19 genera and around 90 recognized species (Fabre et al. 2013) distributed throughout the Neotropics (Emmons and Feer 1997).They are the most diverse group of hystricognath rodents (Leite and Patton 2002) and display a wide range of life histories and ecomorphological adaptations, ranging from semifossorial to arboreal life styles (Emmons and Feer 1997).In Brazil, semifossorial echimyid genera are found mainly in grasslands of the Cerrado and Pantanal (Carterodon and Clyomys) or in the Atlantic Forest ecoregion (Euryzygomatomys) (Paglia et al. 2012;Fabre et al. 2013).Euryzygomatomys spinosus (G.Fischer, 1814) is a medium sized, semifossorial rodent and the only living species currently recognized in its genus (Bonvicino and Bezerra 2015).It is characterized by a fusiform body covered with brown to black spiny dorsal pelage, and short ears and tail (Eisenberg and Redford 1999;Bonvicino et al. 2008), typical of subterranean rodents.The dentition of E. spinosus is unique, characterized by very deep reentrant flexi/flexids, forming elongate fossettes/fossettids, and by the anterior fossette and metafossette subequal in size (Verzi et al. 1995).The karyotype of E. spinosus shows a diploid number (2n) of 46 and fundamental number (FN) of 82 chromosomes (Yonenaga 1975).
This taxon is rare in small mammal surveys (Bonvicino et al. 2002;Scheibler and Christoff 2007;Pinto et al. 2009), leading to relatively few localities with voucher specimens, and therefore resulting in imprecise range boundaries and many distribution gaps.In the present paper, we review published and museum records of E. spinosus and present a distribution map with georeferenced collecting points.We also report the first confirmed occurrence of E. spinosus in the Brazilian Cerrado.

Notes oN GeoGraphic DistributioN
Loss et al. | First record of Euryzygomatomys spinosus in the Cerrado confirm the taxonomic identification, because this molecular marker is an effective tool in species level identification of mammals (Bradley and Baker 2001).Nucleic acids were isolated following Bruford et al. (1992), and the cyt b was amplified by polymerase chain reaction with primers MVZ05 and MVZ16 (Smith and Patton 1993).For a final volume of 25 µl, we added 2.5 µl of buffer (10×), 1.0 µl of MgCl 2 (50 mM), 0.5 µl of deoxynucleoside triphosphate mix (10 mM for each nucleotide), 0.3 ml of each primer (10 mM each), 3 units of Taq Platinum (Invitrogen Corporation, Carlsbad, California), and 1 ml of DNA template.The polymerase chain reaction profiles included an initial denaturation at 94°C for 5 min, followed by 39 cycles and a final extension at 72°C for 7 min.Cycles began with a denaturation at 94°C for 30 s, annealing at 48°C for 45 s, and extension at 72°C for 45 s.The sample was sequenced in both directions with the same primers listed above.The cyt b sequence was aligned using Geneious 5.6 (Biomatters, Auckland, New Zealand), deposited in GenBank (http://www.ncbi.nlm.nih.gov/genbank)under the accession number KM514670, and compared with other sequences from GenBank (accession numbers EU544667, JF297804, U34858) using pairwise p-distance calculated in Geneious.
In order to map the range of E. spinosus, we used records from published papers and museum vouchers.Euryzygomatomys spinosus is the only semifossorial Echimyidae occupying the Atlantic Forest and can be easily identified by its external morphology, so we considered published records within Atlantic Forest domain as correct.Nonetheless, Clyomys or Carterodon altitude varies from 650-1,520 m (Leal et al. 2008).All specimens were collected in a "campo limpo" area, which is part of a mosaic that includes upland mesophytic semideciduous forests (Figures 1 and 2).
The specimens were trapped in Tomahawk traps (300 × 160 × 160 mm) set on the ground and baited with a mixture of banana, sardine oil, corn meal and oat grains.Two voucher specimens (both adult males) were preserved as study skin, skull, post-cranial skeleton, and tissue samples (liver fixed in 96% ethanol), and have been deposited in the small mammal collection at Departamento de Ciências Biológicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Minas Gerais, Brazil, under the catalog numbers MDIA078 and MDIA086.Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio) issued the collecting permit (number 42892-1).
The first 801 base pairs of the mitochondrial cytochrome b gene (cyt b) were sequenced in order to  We examined external and dental morphology of the two voucher specimens of E. spinosus (Figures 3  and 4) and confirmed the taxonomic identification by sequencing the cyt b from one specimen (MDIA078).The sequence from the specimen collected at Sempre Vivas National Park (Figure 5, locality 1) is 1.1% divergent from that of a specimen collected at Biritiba Mirim (Figure 5, locality 16; Genbank EU544667), state of São Paulo, and 0.6% divergent from that of another specimen collected at Sumidouro (Figure 5, locality 11; Genbank U34858), state of Rio de Janeiro.These low genetic divergence values are congruent with the expectations for intraspecific variation within mammal species (Bradley and Baker 2001), and therefore endorse the identification of this specimen as E. spinosus.We took 25 craniodental measurements with a digital caliper and determine age category based on upper cheekteeth worn, following Bezerra and Oliveira (2010) (Table 1).
The literature suggests that E. spinosus occurs mainly in the Atlantic Forest (Bonvicino et al. 1997;Pinto et al. 2009;Bonvicino and Bezerra 2015) and it is found in the southern portions of this ecoregion (Moojen 1952;Bonvicino et al. 2008) in Paraguay, northeastern Argentina (Eisenberg and Redford 1999;Chu et al. 2003;Cirignoli et al. 2011), and from sea level to high-altitude areas in southeastern and southern Brazil (Table 2; Bonvicino and Bezerra 2015).Indeed, the range of E. spinosus nearly matches the geographic distribution of the Atlantic Forest to the south of the Rio Doce (see maps in Bonvicino et al. 2008;Catzeflis et al. 2008;Bonvicino and Bezerra 2015).The type locality of the species is Atyra, southern Paraguay (Bonvicino and Bezerra 2015), within the Chaco Humedo ecoregion (Olson et al. 2001).Although Paglia et al. (2012) reported E. spinosus in the Atlantic Forest, Pampas and Cerrado, no specimen has ever been recorded in a typical Cerrado habitat, but in transitional areas between Cerrado and Atlantic Forest (Figure 5; localities 2, 8, 17 and 19).Some authors recently indicated the occurrence of E. spinosus   The paucity of reliable geographic information on E. spinosus also reflects how little is known about the biology of this species, including behavior and ecology.For example, information on feeding habits is still very scarce.Gonçalves et al. (2007)    habitats and can be considered a habitat generalist (Mares and Ojeda 1982;Bonvicino and Bezerra 2015).This species occurs in Pinus plantations (Gonçalves et al. 2007), montane forests (Bonvicino et al. 1997;Geise et al. 2004), secondary vegetation (Lessa et al. 1999;Scheibler and Christoff 2007), dry grasslands (this study), wet meadows (Stallings 1989) and nearby agriculture fields (D'Andrea et al. 2007;Pinto et al. 2009), from sea level to high altitude areas (above 2,300 m; Table 2).

ACKNOWLEDGEMENTS
We thank Kamila Freitas and Sara Angelica Santos for fieldwork assistance.Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio), regional bureau of Diamantina, Daniel Rios and Jeronimo Martins for their logistical support at Sempre Vivas National Park.Molecular laboratory work was conducted with grant support from Fundação de Amparo à Pesquisa e Inovação do Estado do Espírito Santo (FAPES, grant # 52274322) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, grant # 305008/2013-7) to YLRL.ACL had a predoctoral scholarship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES; Programa de Demanda Social) and has a postdoctoral scholarship from FAPES (grant # 68854315/14).

Figure 1 .
Figure 1.Map showing Sempre Vivas National Park and its vegetation, based on Embrapa Cerrados (2006) and Sano et al. (2007).Red square inside Park area represents collecting locality of the two specimens of Euryzygomatomys spinosus.Names are from municipalities and shaded area in the Brazil inset map corresponds to the Cerrado range.

Figure 4 .
Figure 4. Dorsal and ventral views of the skull of Euryzygomatomys spinosus (MDIA 078) collected at Sempre Vivas National Park, Minas Gerais, Brazil.Photo by Pacheco MAC.Scale bar = 20 mm.
observed the consumption of Pinus taeda (Pinaceae) in southern Brazil andPaglia et al. (2012) andBonvicino and Bezerra (2015) classified E. spinosus as herbivore, whileStallings (1989) considered it as most probably herbivore-grazer.One of us (YLRL) observed E. spinosus consuming leaves of the invasive exotic grass Melinis minutiflora P. Beauv., which dominated the grassland habitat where these rodents lived at Poço das Antas Biological Reserve, state of Rio de Janeiro.The fecal analysis of the specimens collected in this study indicates the consumption of vegetative parts of plants (leaves and steams), but also insects (Coleoptera, Hymenoptera, and Hemiptera).These findings suggest an omnivorous diet in E. spinosus, but this is based on only two adult males.Euryzygomatomys spinosus occupies different

Figure 5 .
Figure 5. Collecting localities of Euryzygomatomys spinosus in Brazil, Paraguay and Argentina.Blue dots represent published records; red dots show new records based on specimens examined from museum collections and published data; red square (locality 1) shows our new record from Sempre Vivas National Park, in the Brazilian Cerrado.Numbers correspond to localities listed in Table2.Green area corresponds to the Atlantic Forest, yellow area to the Cerrado, brown area to the Pampas and orange area to the Chaco Humedo.Map based onOlson et al. (2001) and IBGE (2004).

Table 1 .
Selected measurements (in mm) of the two specimens of Euryzygomatomys spinosus from Sempre Vivas National Park, Minas Gerais, Brazil.

Table 2 .
Confirmed records of Euryzygomatomys spinosus sorted according to numbered localities mapped in Figure5, and corresponding reference and/or voucher number (when examined).References in bold indicate species identity confirmed also by DNA analysis and corresponding Genbank accession code for cyt b sequence in parenthesis.
Loss et al. | First record of Euryzygomatomys spinosus in the Cerrado