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      Three New Species of Phytotelm-Breeding Melanophryniscus from the Atlantic Rainforest of Southern Brazil (Anura: Bufonidae)

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          Abstract

          Three new species of Melanophryniscus are described from the Serra do Mar mountain range of the state of Santa Catarina, southern Brazil. All species are found at intermediate to high altitudes and share phytotelm-breeding as their reproductive strategy. The new species are distinguished from other phytotelm-breeding Melanophryniscus based on different combinations of the following traits: snout-vent length, presence of white and/or yellow spots on forearms, mouth, belly and cloaca, pattern and arrangement of warts, and presence and number of corneous spines. The discovery of these species in a rather restricted geographical area suggests that the diversity of phytotelm-breeding species of Melanophryniscus might be severely underestimated. The conservation status of these species is of particular concern, given that one of them is at risk of extinction not only due to its restricted habitat, but also because of anthropogenic disturbances.

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          Seven new microendemic species of Brachycephalus (Anura: Brachycephalidae) from southern Brazil

          Brachycephalus (Anura: Brachycephalidae) is a remarkable genus of miniaturized frogs of the Brazilian Atlantic Rainforest. Many of its species are highly endemic to cloud forests, being found only on one or a few mountaintops. Such level of microendemism might be caused by their climatic tolerance to a narrow set of environmental conditions found only in montane regions. This restriction severely limits the chance of discovery of new species, given the difficulty of exploring these inaccessible habitats. Following extensive fieldwork in montane areas of the southern portion of the Atlantic Rainforest, in this study we describe seven new species of Brachycephalus from the states of Paraná and Santa Catarina, southern Brazil. These species can be distinguished from one another based on coloration and the level of rugosity of the skin in different parts of their body. These discoveries increase considerably the number of described species of Brachycephalus in southern Brazil.
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            Dietary alkaloid sequestration in a poison frog: an experimental test of alkaloid uptake in Melanophryniscus stelzneri (Bufonidae).

            Several lineages of brightly colored anurans independently evolved the ability to secrete alkaloid-containing defensive chemicals from granular glands in the skin. These species, collectively referred to as 'poison frogs,' form a polyphyletic assemblage that includes some species of Dendrobatidae, Mantellidae, Myobatrachidae, Bufonidae, and Eleutherodactylidae. The ability to sequester alkaloids from dietary arthropods has been demonstrated experimentally in most poison frog lineages but not in bufonid or eleutherodactylid poison frogs. As with other poison frogs, species of the genus Melanophryniscus (Bufonidae) consume large numbers of mites and ants, suggesting they might also sequester defensive alkaloids from dietary sources. To test this hypothesis, fruit flies dusted with alkaloid/nutritional supplement powder were fed to individual Melanophryniscus stelzneri in two experiments. In the first experiment, the alkaloids 5,8-disubstituted indolizidine 235B' and decahydroquinoline were administered to three individuals for 104 days. In the second experiment, the alkaloids 3,5-disubstituted indolizidine 239Q and decahydroquinoline were given to three frogs for 153 days. Control frogs were fed fruit flies dusted only with nutritional supplement. Gas chromatography/mass spectrometry analyses revealed that skin secretions of all experimental frogs contained alkaloids, whereas those of all control frogs lacked alkaloids. Uptake of decahydroquinoline was greater than uptake of 5,8-disubstituted indolizidine, and uptake of 3,5-disubstituted indolizidine was greater than uptake of decahydroquinoline, suggesting greater uptake efficiency of certain alkaloids. Frogs in the second experiment accumulated a greater amount of alkaloid, which corresponds to the longer duration and greater number of alkaloid-dusted fruit flies that were consumed. These findings provide the first experimental evidence that bufonid poison frogs sequester alkaloid-based defenses from dietary sources.
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              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.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2 December 2015
                2015
                : 10
                : 12
                : e0142791
                Affiliations
                [1 ]Programa de Pós-Graduação em Ecologia, Conservação e Manejo da Vida Silvestre, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil
                [2 ]Departamento de Zoologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil
                [3 ]Mater Natura—Instituto de Estudos Ambientais, Curitiba, Paraná, Brazil
                [4 ]Laboratorio de Genética Evolutiva, Instituto de Biología Subtropical, Facultad de Ciencias Exactas Químicas y Naturales, Universidad Nacional de Misiones, Misiones, Argentina
                [5 ]Escola de Saúde, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, Brazil
                [6 ]Programa de Pós-Graduação em Zoologia, Departamento de Zoologia, Universidade Federal do Paraná, Curitiba, Paraná, Brazil
                [7 ]STCP Engenharia de Projetos Ltda, Curitiba, Paraná, Brazil
                Trier University, GERMANY
                Author notes

                Competing Interests: SAAM is employed by STCP Engenharia de Projetos Ltda. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

                Conceived and designed the experiments: MRB LFR MRP. Performed the experiments: MRB CRF DB LFR MRP. Analyzed the data: MRB CRF DB MRP. Contributed reagents/materials/analysis tools: MRB CRF LFR RB-L LC MRP. Wrote the paper: MRB DB. Collected material: MRB LFR RB-L LC SAAM MRP.

                Article
                PONE-D-15-11161
                10.1371/journal.pone.0142791
                4667932
                26630281
                8c2941f2-3816-4ee7-87c4-df78077008ac
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 18 March 2015
                : 27 October 2015
                Page count
                Figures: 16, Tables: 1, Pages: 35
                Funding
                MRB received a PRODOC grant from CAPES (project 2599/2010) and currently receives a PNPD grant from CAPES. RBL is supported by fellowships from CNPq/MCT (141823/2011–9). MRP is supported by a grant from CNPq/MCT (571334/2008–3). DB receives financial support for Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) and ANPCyT (PIP 112201101– 00875, PICT 2007–2202, 2011-1524, 2011–1895, 2012–2687, and 404–2013). Fieldwork during 2011 and 2012 was partially funded by Fundação Grupo O Boticário de Proteção à Natureza (through the project 0895_20111 conducted by Mater Natura - Instituto de Estudos Ambientais). The additional funds were obtained in the form of the other fellowships indicated to some of the authors. STCP Engenharia de Projetos Ltda provided support in the form of salaries for one of the authors (SAAM), but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the “author contributions” section.
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