Calomys tener is a widely distributed species in Brazil. Herein we report the second record of C. tener in state of Rio Grande do Sul about 80 km northwestwards from the previous southernmost known limit in Brazil. A fragment of 890 bp of Cytochrome b gene (cyt b) from MCN -MAM 42 was sequenced and the sequence identity was first investigated by using BLAST (Basic Local Alignment Tool) (Altschul et al. 1990) in order to observe its similarity levels with other sequences deposited in the GenBank. We also performed a Neighbor-Joining (NJ) analysis in MEGA5 (Tamura et al. 2011), which included the cyt b sequence from one specimen here surveyed and specimens from other 10 recognized species of Calomys generated in the studies of Salazar-Bravo et al. (2001;2002) and Almeida et al. (2007), obtained from GenBank ( Figure 4; Appendix 1). The specimen here surveyed grouped with the specimen of C. tener from Goiás, Central Brazil (Almeida et al. 2007) with a high level of bootstrap support (100%). The new cyt b sequence was deposited in GenBank (access number: JX975467). Kimura 2-parameter (K2P) evolutionary distances between Banhado Grande specimen and sequences from the specimens utilized in NJ analysis are shown in Table 1. Cranial measurements (Table 2) were within the range presented by Bonvicino et al. (2003) for C. tener, except by the length of diastema (LD) and breadth of rostrum (BRO). The specimen presented tawny dorsal pelage, lighter on the face; the venter was lighter, but hairs gray-based and with color limits with dorsum less conspicuous when compared to sympatric C. laucha. White hairs behind ears also distinguished these two species in the region, being a conspicuous characteristic in C. laucha and less conspicuous in C. tener. This is the second record of Calomys tener for the state of Rio Grande do Sul. Although it not extends its distribution, we confirm the presence of this species in a region approximately 80 km northwestwards from the locality of Quintão, the previous southernmost known limit in Rio Grande do Sul years of small mammal surveys conducted in different physiognomies of the state (e.g. Cademartori et al. 2002; 2004; Dalmagro and Vieira 2005;Iob and Vieira 2008;Quintela et al. 2012;Sponchiado et al. 2012) and the record in only two localities until now (Haag et al. 2007 and this study). Calomys laucha, on the other hand, seems to be a common species in southern coastal dune systems (e.g. Camargo et al. 2006; Haag et al. 2007;Quintela et al. 2013), with one record that may represents its northernmost limit of distribution in a shrubby area in the north region of the state (Badzinski et al. 2012). Since this is the first record of sympatry between these two morphologically similar species in the genus, the real distribution limits MCN-MAM 42
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