Distributed under Creative Commons Cc-by 4.0 a New Species of Brachycephalus (anura: Brachycephalidae) from the Quiriri Mountain Range of Southern Brazil

A new miniaturized toadled of the genus Brachycephalus (Anura: Brachycephalidae) is described from Serra do Quiriri in the municipality of Campo Alegre, Santa Catarina, southern Brazil. Specimens were collected from the leaf litter between from 1,263 and 1,318 m above sea level. The new species is distinguished from all its congeners by the combination of the following characters: snout–vent length 9.9–13.1 mm; skin on head and dorsum without dermal co-ossification; snout mucronate in dorsal view; dorsum rugose; general color brown, with a narrow orange vertebral stripe. The region where the new species is located is also shared with other endemic anuran species and has experienced strong anthropogenic impacts,suggesting that immediate actions should be taken to ensure their long-term preservation.


ghout the BAF
(e.g., Da Silva, Cardoso de Sousa & Castelletti, 2004;Sigrist & Carvalho, 2008), possibly as a result of the complex evolutionary history of the biome as it responded to climatic changes over the course of the Quaternary (Carnaval et al., 2009;Carnaval et al., 2014).Some taxa endemic to the BAF show extremely reduced geographical ranges, most commonly in association with montane regions, such as many species of the miniaturized toadlet Brachycephalus Fitzinger, 1826 (Anura: Brachycephalidae) (Pombal, Wistuba & Bornschein, 1998;Ribeiro et al., 2005;Alves et al., 2006;Napoli et al., 2011;Pie et al., 2013).Despite the intriguing evolutionary mechanisms underlying the diversification of Brachycephalus, only recently have their phylogenetic relationships begun to be investigated (e.g., Clemente-Carvalho et al., 2011;Padial, Grant & Frost, 2014).

The species of Brachycephalus have been recently divided into three species groups (Ribeiro et al., 2015): ephippium (Spix, 1824), didactylus (Izecksohn, 1971), andpernix Pombal, Wistuba &Bornschein, 1998.The latter included the species of Brachycephalus that shared th ir bufoniform body shape with the ephippium group, but that lacked their characteristic dermal ossification, namely B. pernix Pombal, Wistuba & Bornschein, 1998, B. brunneus Ribeiro, Alves, Haddad & Reis, 2005, B. izecksohni Ribeiro, Alves, Haddad & Reis, 2005, B. ferruginus Alves, Ribeiro, Haddad & Reis, 2006, B. pombali Alves, Ribeiro, Haddad & Reis, 2006, and B. tridactylus Garey, Lima, Hartmann & Haddad, 2012.However, a single additional study more than doubled the number of species in the pernix group (Ribeiro et al., 2015), suggesting that the actual diversity in the group could be greatly underestimated.In the present study, we describe a new species of the pernix group of Brachycephalus from the Quiriri mountain range of the state of Santa Catarina, southern Brazil.In addition to clear morphological diagnostic traits, the new species tends to be isolated from one another by valleys of unsuitable habitats, essentially forming sky islands (Howell, 1947;see McCormack, Huang & Knowles, 2009 for a recent review).


MATERIALS AND METHODS

Adult specimens were anaesthetized and euthanized using 2% lidocaine, fixed in 10% formalin, and preserved in 70% ethanol.Measurements were made with a micrometric eyepiece attached to a stereomicroscope.Measurements abbreviations were as follows:

nout-vent length (SVL)
head length, from tip of snout to angle of jaw (HL); head width-greatest width of head located between angles of jaw (HW); eye diameter (ED); nostril diameter (ND); interorbital distance between anterior corners of the eyes (IOD); internostril distance between inner margins of nostrils (IND); eye-nostril distance from anterior corner of the eye to posterior margin of nostril (END); thigh length (THL); and tibia length (TBL).

All specimens collected for this study were deposited in the Colec ¸ão Herpetol ógica do Departamento de Zoologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil (DZUP).In addition, we examined specimens from the following collections: Célio F. B. Haddad collecti n, deposited in the Departamento de Zoologia, Universidade Estadual Paulista, Campus de Rio Claro, São Paulo, Brazil (CFBH), DZUP, Museu de Hist ória Natural Capão da Imbuia, Curitiba, Paraná, Brazil (MHNCI), Museu Nacional, Rio de Janeiro, Rio de Janeiro, Brazil (MNRJ), Museu de Zoologia da Universidade de São Paulo, São Paulo, São Paulo, Brazil (MZUSP), and Museu de Hist ória Natural, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil (ZUEC).A list of the examined specimens is provided in Appendix S1.In this study we adopted the evolutionary species concept, which equates species with "separately evolving (segments of) metapopulation lineages" (De Queiroz, 2005), which is particularly suitable for the evolutionary scenario of population isolation in sky islands followed by divergent evolution.

The electronic version of this article in Portable Document Format (PDF) will represent a published work according to the International Commission on Zoological Nomenclature (ICZN), and hence the new name contained in the electronic version is effectively published under that Code from the electronic edition alone.This published work and the nomenclatural act it contains has been registered in ZooBank, the online registration system for the ICZN.The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix "http://zoobank.org/."The LSID for this publication is: 5267B6A8-373D-4DD2-9F7F-9D41FB6933E0.The online version of this work is archived and available from the following digital repositories: PeerJ, PubMed Central and CLOCKSS.

Collection permits for this study were issued by ICMBIO (SISBIO 1911426).We classified vegetation in collection sites according to the classification of Veloso, Rangel-Filho & Lima (1991).All coordinates in this study were based on datum WGS84.


RESULTS

Brachycephalus quir riensis sp.nov.Urn:lsid:zoobank.org:act:A58C8AC5-6C38-447E-B2B2-3BED619FEFC8 (Figs. 1, 2 and 3) Brachycephalus sp. 5 (Pie et al., 2013) Holotype.DZUP 172 (Fig. 1), adult female, collected at the Serra do Quiriri, ( 26• 01 ′ 17 ′′ S, 48 Diagnosis.B

chycepha
us quiririensis is distinguished from all of the species in the genus by the following combination of characters: body robust, bufoniform, adult size SVL 9.9-13.1 mm, rough dorsum (Fig. 1); general background color greenish-brown, with an orange stripe that extends from the dorsum of the head and along the vertebral column; skin on dorsum of head and central body dorsum with no dermal co-ossification (Fig. 2).A representative of the pernix group, B. quiririensis is unique among other members of its species group by its brownish dorsal coloration shared B. brunneus, but different from that species due to the orange stripe along the vertebral column and by the greenish hue associated with the brown coloration.vocal sac not expanded externally; vocal slits present; tongue longer than wide, with posterior half not adherent to floor of mouth; choanae relatively small and round; vomerine odonthophores absent; arm and forearm relatively slender; arm approximately as long as forearm; tip of finger I and II slightly rounded, tip of finger III pointed; finger I and IV very small, vestigial; relative lengths of fingers IV <I < II < III; subarticular tubercles absent; inner and outer metacarpal tubercles absent; legs short, moderately robust; thigh length 34% of snout-vent length, tibia length 79 % of thigh length; toes II-III short, relatively distinct; toe I externally absent, to V vestigial; relative length of toes V < II < III < IV; subarticular tubercles and inner metatarsal tubercles absent; outer metatarsal tubercle distinct, large, ovoid; rough dorsum, without co-ossifications; glandular warts large, circular to oval, present and spaced in dorsum and legs; head and arms smooth; sides of the body granular; large glandular warts circular and spaced on the sides of the body; belly granular; circular glandular warts spaced and distributed from belly to legs and arms, chin smooth.Coloration of Holotype.In life, dorsum, head, sides of the body, dorsal region of arms, thighs, and legs brown; dorsal region of snout and head orange, continuing into an orange stripe along the vertebral column and forming two spots near the tips of the sacral vertebra; chin, ventral portion of arms, thighs, legs, and ventral regions of hand and feet orange; belly with orange spots against a the brownish coloration that extends from the sides of the body; iris black (Fig. 2).In preservative, the orange coloration becomes pale cream, while maintaining the brownish coloration.Variation in Type Series.Measurements and proportions of 10 adults in millimeters are (mean ± SD, with range in parentheses): SVL 11.12 ± 1.03 (9.9-13.1);HL 3.73 ± 0.31 (3.3-4.4);HW 4.23 ± 0.42 (3.7-5.0);ED 1.18 ± 0.10 (1.1-1.4);ND 0.21 ± 0.04 (0.2-0.3); IOD 2.25 ± 0.17 (2.0-2.6);IND 1.26 ± 0.11 (1.1-1.4);END 0.64 ± 0.07 (0.6-0.8); THL 3.83 ± 0.37 (3.1-4.7);TBL 3.23 ± 0.30 (2.9-3.8);THL/SVL 35 ± 2% (32-37%); TBL/THL 85 ± 6% (70-93%); HL/SVL 34 ± 1% (31-36%); ED/HL 32 ± 2% (28-34%); HL/HW 88 ± 5% (79-93%).There is variation am

g individuals in the bre
dth of the orange stripe along their backs, as well as different degrees of darkness in the brown regions (Fig. 3).Some specimens show a slight greenish hue in the brownish regions of the body.


Measurements of

Etymology.The epithet "quiririensis" is derived from the Tupi-Guarani language word "quiriri" (= silence, peace) and refers to type locality.

Distribution.Brachycephalus quiririensis is known from the type locality, as well as an additional nearby site (Fig. 4) in the municipality of Garuva, state of Santa Catarina (26 • 01 ′ 42 ′′ S, 48 • 57 ′′ 11 ′′ W) (Pie et al., 2013).

Remarks.Individuals of B. quiririensis were found on leaf litter of a cloud forest ("Floresta Ombr ófila Densa Altomontana" sensu Veloso, Rangel-Filho & Lima, 1991) from 1,263 m at 1,318 m a.s.l.During two visits to the type locality, we found individuals of B. quiririensis hidden in the leaf litter, some of which vocalizing.The vegetation in the area has experienced severe disturbances in recent years, particularly fire and extensive cattle ranching.Moreover, two highly endemic bufonid species of the genus Melanophryniscus are found in the region (Pie et al., 2013;Bornschein et al., 2015, unpublished data), one of which occurring syntopically with B. quiririensis.Strong measures should be taken by the environmental protect

n agency of the
tate of Santa Catarina to ensure the long-term preservation of these species.In addition, B. quiririensis should be integrated into the "Plano

Ac ¸ão Nacion
l para a Conservac ¸ão dos Anfíbios e Répteis Ameac ¸ados da Região Sul do Brasil," an initiative organized by the Instituto Ch co Mendes/Ministry of the Environment that seeks to organize conservation initiatives for the conservation of the herpetofauna of southern Brazil.

Figure 1
1
Figure 1 Holotype of Brachycephalus quiririensis (DZUP 172).(A) Dorsal view of the body, (B) ventral view of the body, (C) lateral view of the head, (D) ventral view of right hand, (E) ventral view of right foot.Horizontal scale bar = 5 mm; vertical scale bars = 2 mm.


Figure 2
2
Figure 2 Brachycephalus quiririensis in life.




Holotype.SVL = 12.3 mm, HL = 3.9 mm, HW = 4.7 mm, ED = 1.3 mm, ND = 0.3 mm, IOD = 2.4 mm, IND = 1.3 mm, END = 0.7 mm, THL = 4.1 mm, TBL = 3.3 mm.


Figure 3 Figure 4
34
Figure 3 Variation in dorsal coloration in live (anesthetized) specimens of Brachycephalus quiririensis.




The new species is also distinguished from B. ferruginus,B.izecksohni,B. pombali, and B.tridactylus by having rough dorsum, with granular aspect, instead of a smooth dorsum.The new species lacks of dermal co-ossification characteristic of species of the ephippium group, and the bufoniform shape and larger body size of the new species distinguish it from those in the didactylus group, which are smaller (SVL = 8-10 m