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      First record of Vitreorana ritae (Anura, Centrolenidae) for southern Amazonia inferred from molecular, reproductive and acoustic evidence Translated title: Primer registro de Vitreorana ritae (Anura, Centrolenidae) en el sur de la Amazonia inferida a partir de evidencia molecular, reproductiva y acústica

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          Abstract

          ABSTRACT Frogs of the family Centrolenidae are adapted to live at high altitudes, and so it is not surprising that their greatest diversity is in the Andes mountain range. Here we extend the known geographic distribution of Vitreorana ritae by more than 550 km towards southern Amazonia. The species was identified by morphological, acoustic and molecular characteristics (mitochondrial marker 16S rRNA). The records reported here are the first for the genus Vitreorana in the state of Mato Grosso, Brazil, as well as for the transition zone between the Amazonia and Cerrado biomes. These records are consistent with the hypothesis that the low number of species of centrolenids reported in the lowlands of the Amazon region may be the result of limited sampling. Even though V. ritae is distributed throughout the Amazon, most of its diagnostic morphological characteristics are conserved. Thus, the great rivers of the Amazon Basin do not seem to act as geographical barriers for this species; however, due to the limited sample size, further acoustic and molecular studies are needed to confirm this hypothesis.

          Translated abstract

          RESUMEN Las especies de anfibios de la familia Centrolenidae están adaptadas a vivir en regiones de elevada altitud, por lo que no sorprende que su mayor diversidad ocurra en la Cordillera de los Andes. Aquí expandimos el ámbito de distribución de Vitreorana ritae por más de 550 km al sur del río Amazonas. La identificación de la especie se realizó con base en la comparación de las características morfológicas, acústicas y moleculares (marcador mitocondrial 16S rRNA). Nuestro registro es el primero de este género para el Estado de Mato Grosso, Brasil, así como para la zona de transición entre los biomas Amazonía y Cerrado. Este registro es concordante con la hipótesis de que el bajo número de especies registradas en la planicie amazónica puede ser el resultado de un menor esfuerzo de muestreo. Observamos que, a pesar de la amplia distribución de la especie en la Amazonía, la mayoría de las características morfológicas diagnósticas de V. ritae se conservan. Sugerimos que los grandes ríos de la cuenca amazónica parecen no actuar como barrera geográfica para esta especie. Sin embargo, y como resultado de nuestro pequeño tamaño de muestra, se requerirán estudios acústicos y moleculares adicionales para confirmar esta hipótesis.

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          Most cited references42

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          MEGA6: Molecular Evolutionary Genetics Analysis version 6.0.

          We announce the release of an advanced version of the Molecular Evolutionary Genetics Analysis (MEGA) software, which currently contains facilities for building sequence alignments, inferring phylogenetic histories, and conducting molecular evolutionary analysis. In version 6.0, MEGA now enables the inference of timetrees, as it implements the RelTime method for estimating divergence times for all branching points in a phylogeny. A new Timetree Wizard in MEGA6 facilitates this timetree inference by providing a graphical user interface (GUI) to specify the phylogeny and calibration constraints step-by-step. This version also contains enhanced algorithms to search for the optimal trees under evolutionary criteria and implements a more advanced memory management that can double the size of sequence data sets to which MEGA can be applied. Both GUI and command-line versions of MEGA6 can be downloaded from www.megasoftware.net free of charge.
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            The use of bioacoustics in anuran taxonomy: theory, terminology, methods and recommendations for best practice.

            Vocalizations of anuran amphibians have received much attention in studies of behavioral ecology and physiology, but also provide informative characters for identifying and delimiting species. We here review the terminology and variation of frog calls from a perspective of integrative taxonomy, and provide hands-on protocols for recording, analyzing, comparing, interpreting and describing these sounds. Our focus is on advertisement calls, which serve as premating isolation mechanisms and, therefore, convey important taxonomic information. We provide recommendations for terminology of frog vocalizations, with call, note and pulse being the fundamental subunits to be used in descriptions and comparisons. However, due to the complexity and diversity of these signals, an unequivocal application of the terms call and note can be challenging. We therefore provide two coherent concepts that either follow a note-centered approach (defining uninterrupted units of sound as notes, and their entirety as call) or a call-centered approach (defining uninterrupted units as call whenever they are separated by long silent intervals) in terminology. Based on surveys of literature, we show that numerous call traits can be highly variable within and between individuals of one species. Despite idiosyncrasies of species and higher taxa, the duration of calls or notes, pulse rate within notes, and number of pulses per note appear to be more static within individuals and somewhat less affected by temperature. Therefore, these variables might often be preferable as taxonomic characters over call rate or note rate, which are heavily influenced by various factors. Dominant frequency is also comparatively static and only weakly affected by temperature, but depends strongly on body size. As with other taxonomic characters, strong call divergence is typically indicative of species-level differences, whereas call similarities of two populations are no evidence for them being conspecific. Taxonomic conclusions can especially be drawn when the general advertisement call structure of two candidate species is radically different and qualitative call differences are thus observed. On the other hand, quantitative differences in call traits might substantially vary within and among conspecific populations, and require careful evaluation and analysis. We provide guidelines for the taxonomic interpretation of advertisement call differences in sympatric and allopatric situations, and emphasize the need for an integrative use of multiple datasets (bio-acoustics, morphology, genetics), particularly for allopatric scenarios. We show that small-sized frogs often emit calls with frequency components in the ultrasound spectrum, although it is unlikely that these high frequencies are of biological relevance for the majority of them, and we illustrate that detection of upper harmonics depends also on recording distance because higher frequencies are attenuated more strongly. Bioacoustics remains a prime approach in integrative taxonomy of anurans if uncertainty due to possible intraspecific variation and technical artifacts is adequately considered and acknowledged.
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              Underestimation of Species Richness in Neotropical Frogs Revealed by mtDNA Analyses

              Background Amphibians are rapidly vanishing. At the same time, it is most likely that the number of amphibian species is highly underestimated. Recent DNA barcoding work has attempted to define a threshold between intra- and inter-specific genetic distances to help identify candidate species. In groups with high extinction rates and poorly known species boundaries, like amphibians, such tools may provide a way to rapidly evaluate species richness. Methodology Here we analyse published and new 16S rDNA sequences from 60 frog species of Amazonia-Guianas to obtain a minimum estimate of the number of undescribed species in this region. We combined isolation by distance, phylogenetic analyses, and comparison of molecular distances to evaluate threshold values for the identification of candidate species among these frogs. Principal Findings In most cases, geographically distant populations belong to genetically highly distinct lineages that could be considered as candidate new species. This was not universal among the taxa studied and thus widespread species of Neotropical frogs really do exist, contrary to previous assumptions. Moreover, the many instances of paraphyly and the wide overlap between distributions of inter- and intra-specific distances reinforce the hypothesis that many cryptic species remain to be described. In our data set, pairwise genetic distances below 0.02 are strongly correlated with geographical distances. This correlation remains statistically significant until genetic distance is 0.05, with no such relation thereafter. This suggests that for higher distances allopatric and sympatric cryptic species prevail. Based on our analyses, we propose a more inclusive pairwise genetic distance of 0.03 between taxa to target lineages that could correspond to candidate species. Conclusions Using this approach, we identify 129 candidate species, two-fold greater than the 60 species included in the current study. This leads to estimates of around 170 to 460 frog taxa unrecognized in Amazonia-Guianas. Significance As a consequence the global amphibian decline detected especially in the Neotropics may be worse than realised.
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                Author and article information

                Journal
                cal
                Caldasia
                Caldasia
                Instituto de Ciencias Naturales, Facultad de Ciencias-Universidad Nacional de Colombia (Bogotá, Cundinamarca, Colombia )
                0366-5232
                2357-3759
                December 2020
                : 42
                : 2
                : 171-180
                Affiliations
                [2] orgnameUniversidade Federal de Mato Grosso Brazil
                [6] Goiás orgnameUniversidade Federal de Goiás Brazil
                [5] Pará orgnameUniversidade Federal do Oeste do Pará Brazil
                [3] Amazonas orgnameUniversidade Federal do Amazonas Brazil
                [4] Pará orgnameUniversidade Federal do Pará Brazil
                [1] orgnameUniversidade Federal de Mato Grosso Brazil penhacek@ 123456yahoo.com.br
                Article
                S0366-52322020000200171 S0366-5232(20)04200200171
                10.15446/caldasia.v42n2.79487
                c9e1fed1-7abc-4471-a454-8fd6d87b6a97

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 24 May 2020
                : 09 July 2019
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 58, Pages: 10
                Product

                SciELO Colombia


                Amazon-Cerrado ecotone,intraspecific genetic variation,variación genética intraespecífica,Acoustic parameters,rana de cristal,glass frog,parámetros acústicos,Ecótono Amazonía-Cerrado

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