Effects of the insecticide imidacloprid on aquatic invertebrate communities of the Ecuadorian Amazon.

Imidacloprid is a neonicotinoid insecticide that has received particular attention due to its widespread use and potential adverse effects for aquatic and terrestrial ecosystems. Its toxicity to aquatic organisms has been evaluated in central and southern Europe as well as in (sub-)tropical regions of Africa and Asia, showing high toxic potential for some aquatic insects and zooplankton taxa. However, its toxicity to aquatic organisms representative of tropical regions of Latin America has never been evaluated. To fill this knowledge gap, we carried out a mesocosm experiment to assess the short- and long-term effects of imidacloprid on freshwater invertebrate communities representative of the Ecuadorian Amazon. A mesocosm experiment was conducted with five weekly applications of imidacloprid at four nominal concentrations (0.01 µg/L, 0.1 µg/L, 1 µg/L and 10 µg/L). Toxic effects were evaluated on zooplankton and macroinvertebrate populations and communities, as well as on water quality parameters for 70 days. Given the climatic conditions prevailing in the study area, characterized by a high solar radiation and abundant rainfall that resulted in mesocosm overflow, there was a rapid dissipation of the test compound from the water column (half-life: 4 days). The macroinvertebrate taxa Callibaetis pictus (Ephemeroptera), Chironomus sp. (Diptera), and the zooplankton taxon Macrocyclops sp., showed population declines caused by the imidacloprid treatment, with a 21-d Time Weighted Average No Observed Effect Concentrations (21-d TWA NOEC) of 0.46 µg/L, except for C. pictus which presented a 21-d TWA NOEC of 0.05 µg/L. In general terms, the sensitivity of these taxa to imidacloprid was greater than that reported for surrogate taxa in temperate zones and similar to that reported in other (sub-)tropical regions. These results confirm the high sensitivity of tropical aquatic invertebrates to this compound and suggest the need to establish regulations for the control of imidacloprid contamination in Amazonian freshwater ecosystems.

Field-based molecular detection of Batrachochytrium dendrobatidis in critically endangered Atelopus toads and aquatic habitats in Ecuador.

Batrachochytrium dendrobatidis (Bd) is a lethal fungal species that parasitizes vertebrates and is associated with the worldwide decline of amphibian populations. The development of sensitive, rapid detection methods, particularly DNA-based techniques, is critical for effective management strategies. This study evaluates the efficacy of DNA extraction and a portable PCR device in a mountable field laboratory setup for detecting Bd near the habitats of three critically endangered Atelopus toad species in Ecuador. We collected skin swabs from Atelopus balios, A. nanay, and A. bomolochos, and environmental DNA (eDNA) samples from streams in Andean and coastal regions of Ecuador. For eDNA, a comparison was made with duplicates of the samples that were processed in the field and in a standard university laboratory. Our findings revealed Bd detection in eDNA and swabs from 6 of 12 water samples and 10 of 12 amphibian swab samples. The eDNA results obtained in the field laboratory were concordant with those obtained under campus laboratory conditions. These findings highlight the potential of field DNA-based monitoring techniques for detecting Bd in amphibian populations and their aquatic habitats, particularly in remote areas. Furthermore, this research aligns with the National Action Plan for the Conservation of Ecuadorian Amphibians and contributes to the global effort to control this invasive and deadly fungus.

Environmental DNA and visual encounter surveys for amphibian biomonitoring in aquatic environments of the Ecuadorian Amazon.

Background: The development of anthropogenic activities has generated a decline in aquatic fauna populations, and amphibians have been the most affected. The decline of batrachofauna is concerning, as 41% of all species worldwide are endangered. For this reason, rapid, efficient, and non-invasive biodiversity monitoring techniques are needed, and environmental DNA (eDNA) is one such tool that has been sparsely applied in Ecuador. This technique has allowed scientists generates information on species diversity and amphibian community composition from a water sample. This study applied eDNA-based biomonitoring analyses and visual encounter surveys (VES) as inventory techniques to identify the diversity of aquatic amphibians in the Tena River micro-basin (TRMB). Methods: The experimental design was divided into three components: (1) fieldwork: all amphibians were recorded by the VES technique and water samples were collected; (2) laboratory work: DNA isolation from amphibian tissue samples and eDNA-containing filters, amplification, electrophoresis, and sequencing were performed; (3) Data analysis: a local DNA reference database was constructed, and eDNA sequence data were processed for classification, taxonomic assignment, and ecological interpretation. Results: Using both eDNA and VES, we detected 33 amphibian species (13 with eDNA only, five with VES only, and 15 with both methods). These species belonged to six amphibian families: Hylidae being the richest with 14 species (three eDNA, one VES, and 10 with both methods), followed by Strabomantidae with nine species (six eDNA, one VES, and two with both methods). All families were detected with both methods, except for the Aromobatidae, having one single record (Allobates aff. insperatus) by VES. Individually, eDNA detected 28 species and had a detection probability (DP) of 0.42 CI [0.40-0.45], while VES recorded 20 species with a DP of 0.17 CI [0.14-0.20]. Similarly, using VES, Cochranella resplendens was detected for the first time in TRMB, while with eDNA, four mountain frogs Pristimantis acerus, Pristimantis eriphus, Pristimantis mallii, and Pristimantis sp. (INABIO 15591) previously recorded at 1,518 m.a.s.l. at altitudes below 600 m.a.s.l. were detected. Conclusions: Results obtained in this study showed that eDNA-based detection had a greater capacity to detect amphibians in aquatic environments compared to VES. The combination of VES and eDNA improves the sensitivity of species detection and provides more reliable, robust, and detailed information. The latter is essential for developing conservation strategies in the Ecuadorian Amazon.

Respuesta ecológica multitaxón a la pérdida de hábitat y fragmentación del paisaje en la Amazonía occidental reveladas por evaluaciones de biodiversidad RAPELD / Multi-taxa ecological responses to habitat loss and fragmentation in western Amazonia as revealed by RAPELD biodiversity surveys

La pérdida y fragmentación del hábitat causada por la deforestación es un importante impulsor antropogénico de cambios sobre la biodiversidad en la selva amazónica. Sin embargo, la magnitud y dirección de los efectos sobre la composición y distribución de las especies aún es incomprendida. Evaluamos las respuestas de cuatro grupos taxonómicos - aves, anfibios, abejas de orquídeas y escarabajos peloteros - a la pérdida y fragmentación del hábitat, tanto a nivel de especies como de ensamblaje, en la Amazonía norte ecuatoriana. Tomamos muestras de quince parcelas de 250 m de largo en remanentes de bosque de tierra firme. Calculamos un índice de fragmentación del paisaje (fragindex), que considera la cobertura forestal continua, densidad del borde y el aislamiento en el paisaje, y nueve métricas de configuración del paisaje para analizar las respuestas de especies y ensamblajes. Más del 80% de las especies de aves, anfibios o abejas de orquídeas y el 60% de los escarabajos peloteros se vieron afectados negativamente por la pérdida y fragmentación del hábitat. La composición por especies se vio significativamente afectada por las diferencias en la cobertura forestal y la conectividad, mientras que la forma y el área de los parches de bosque determinaron la magnitud y la dirección del efecto en las respuestas de las especies. Por lo tanto, los cambios en la configuración del paisaje de la Amazonía ecuatoriana deben minimizarse para disminuir los efectos de la pérdida y fragmentación del hábitat sobre la presencia de especies y la composición de los ensambles. (AU)

Red List assessment of amphibian species of Ecuador: A multidimensional approach for their conservation.

Ecuador is one of the most biodiverse countries in the world, but faces severe pressures and threats to its natural ecosystems. Numerous species have declined and require to be objectively evaluated and quantified, as a step towards the development of conservation strategies. Herein, we present an updated National Red List Assessment for amphibian species of Ecuador, with one of the most detailed and complete coverages for any Ecuadorian taxonomic group to date. Based on standardized methodologies that integrate taxonomic work, spatial analyses, and ecological niche modeling, we assessed the extinction risk and identified the main threats for all Ecuadorian native amphibians (635 species), using the IUCN Red List Categories and Criteria. Our evaluation reveals that 57% (363 species) are categorized as Threatened, 12% (78 species) as Near Threatened, 4% (26 species) as Data Deficient, and 27% (168 species) as Least Concern. Our assessment almost doubles the number of threatened species in comparison with previous evaluations. In addition to habitat loss, the expansion of the agricultural/cattle raising frontier and other anthropogenic threats (roads, human settlements, and mining/oil activities) amplify the incidence of other pressures as relevant predictors of ecological integrity. Potential synergic effects with climate change and emergent diseases (apparently responsible for the sudden declines), had particular importance amongst the threats sustained by Ecuadorian amphibians. Most threatened species are distributed in montane forests and paramo habitats of the Andes, with nearly 10% of them occurring outside the National System of Protected Areas of the Ecuadorian government. Based on our results, we recommend the following actions: (i) An increase of the National System of Protected Areas to include threatened species. (ii) Supporting the ex/in-situ conservation programs to protect species considered like Critically Endangered and Endangered. (iii) Focalizing research efforts towards the description of new species, as well as species currently categorized as Data Deficient (DD) that may turn out to be threatened. The implementation of the described actions is challenging, but urgent, given the current conservation crisis faced by amphibians.

Rediscovery of Laura's glassfrog Nymphargus laurae (Anura: Centrolenidae) with new data on its morphology, colouration, phylogenetic position and conservation in Ecuador.

We report the rediscovery of Laura's Glassfrog, Nymphargus laurae Cisneros-Heredia & McDiarmid, 2007, based on two specimens collected at the Colonso-Chalupas Biological Reserve, province of Napo, Ecuador. The species was described and known from a single male specimen collected in 1955 at Loreto, north-eastern Andean foothills of Ecuador. Limited information was available about the colouration, systematics, ecology, and biogeography of N. laurae. We provide new data on the external morphology, colouration, distribution and comment on its conservation status and extinction risk. We discuss the phylogenetic relationships of N. laurae, which forms a clade together with N. siren and N. humboldti. The importance of research in unexplored areas must be a national priority to document the biodiversity associated, especially in protected areas.

A new species of Leucostethus (Anura: Dendrobatidae) from the Cordillera Mache-Chindul in northwestern Ecuador, with comments on similar Colostethus and Hyloxalus.

We describe a new species of dendrobatid frog, Leucostethus bilsa sp. nov., using molecular, morphological, and acoustic evidence. We also comment on the taxonomic status of four similar Hyloxalus and Colostethus. We provide an updated phylogeny of Leucostethus that corroborates previous hypotheses of relationships of nine species. Phylogenetic analysis using mitochondrial (i.e., 7095 bp of combined data from NADH1, NAHD2, cytochrome c oxidase I, cytochrome b and 12S-16S rRNA) and seven nuclear genes (i.e., 4739 bp) indicate a close relationship of L. bilsa to an undescribed species from Gorgona Island, Colombia, both of which apparently diverged in the Pliocene about 3 million years ago with about 6.25% (i.e., 146/2335 bp) differences for the section of 12S-16S mitochondrial fragment. Leucostethus bilsa is diurnal and riparian, characterized by distinctive bright mustard-yellow flash marks in the axillar and groin regions, posterior belly, and in the hindlimbs, the presence of dark gray lower labial stripe or marks, sexual dimorphism in ventral pattern, and by having male uniparental care. We describe its osteology and the male advertisement call, which is a series of peep notes. Osteological microCT images of representatives of each of the Colostethinae genera reveal a number of intriguing characters that may prove to be useful in phylogenetic studies. In terms of its distribution, Leucostethus bilsa is currently known only from a very small area within the Reserva Biológica Bilsa, located within the Cordillera Mache-Chindul in the Chocoan region of northwestern Ecuador, which was a Pliocene-Pleistocene refugium. This region is highly threatened with habitat degradation and remains as the last surviving refuge for a forest community known for a high proportion of endemic species of both flora and fauna.

Systematics of Ecnomiohyla tuberculosa with the description of a new species and comments on the taxonomy of Trachycephalus typhonius (Anura, Hylidae).

Ecnomiohyla tuberculosa is an Amazonian hylid of uncertain phylogenetic position. Herein DNA sequences of mitochondrial and nuclear genes are used to determine its phylogenetic relationships. New sequences and external morphology of Trachycephalus typhonius are also analyzed to assess the status of Ecuadorian and Peruvian populations. The phylogeny shows unequivocally that Ecnomiohyla tuberculosa is nested within the genus Tepuihyla, tribe Lophiohylini. This position was unexpected because the remaining species of Ecnomiohyla belong to the tribe Hylini. To solve the paraphyly of the genus Ecnomiohyla, Ecnomiohyla tuberculosa is transferred to the genus Tepuihyla. Comparisons of DNA sequences, external morphology, and advertisement calls between populations of Ecnomiohyla tuberculosa from Ecuador and Peru indicate that the Peruvian population represents an undescribed species. The new species is described and a species account is provided for Ecnomiohyla tuberculosa. Trachycephalus typhonius is paraphyletic relative to Trachycephalus cunauaru, Trachycephalus hadroceps, and Trachycephalus resinifictrix. The phylogenetic position of populations from western Ecuador indicates that they represent a species separate from Trachycephalus typhoniussensu stricto. We resurrect the name Hyla quadrangulum (Trachycephalus quadrangulumcomb. n.) for those populations. Amazonian populations of "Trachycephalus typhonius" from Ecuador and Peru are genetically and morphologically distinct from Trachycephalus typhoniussensu stricto and are conspecific with the holotype of Hyla macrotis. Therefore, we also resurrect Hyla macrotis, a decision that results in Trachycephalus macrotiscomb. n.

Insights from Integrative Systematics Reveal Cryptic Diversity in Pristimantis Frogs (Anura: Craugastoridae) from the Upper Amazon Basin.

Pluralistic approaches to taxonomy facilitate a more complete appraisal of biodiversity, especially the diversification of cryptic species. Although species delimitation has traditionally been based primarily on morphological differences, the integration of new methods allows diverse lines of evidence to solve the problem. Robber frogs (Pristimantis) are exemplary, as many of the species show high morphological variation within populations, but few traits that are diagnostic of species. We used a combination of DNA sequences from three mitochondrial genes, morphometric data, and comparisons of ecological niche models (ENMs) to infer a phylogenetic hypothesis for the Pristimantis acuminatus complex. Molecular phylogenetic analyses revealed a close relationship between three new species-Pristimantis enigmaticus sp. nov., P. limoncochensis sp. nov. and P. omeviridis sp. nov.-originally confused with Pristimantis acuminatus. In combination with morphometric data and geographic distributions, several morphological characters such as degree of tympanum exposure, skin texture, ulnar/tarsal tubercles and sexual secondary characters (vocal slits and nuptial pads in males) were found to be useful for diagnosing species in the complex. Multivariate discriminant analyses provided a successful classification rate for 83-100% of specimens. Discriminant analysis of localities in environmental niche space showed a successful classification rate of 75-98%. Identity tests of ENMs rejected hypotheses of niche equivalency, although not strongly because the high values on niche overlap. Pristimantis acuminatus and P. enigmaticus sp. nov. are distributed along the lowlands of central-southern Ecuador and northern Peru, in contrast with P. limoncochensis sp. nov. and P. omeviridis sp. nov., which are found in northern Ecuador and southern Colombia, up to 1200 m in the upper Amazon Basin. The methods used herein provide an integrated framework for inventorying the greatly underestimated biodiversity in Amazonia.

Ecological and geographical analysis of the distribution of the mountain tapir (Tapirus pinchaque) in Ecuador: importance of protected areas in future scenarios of global warming.

In Ecuador, Tapirus pinchaque is considered to be critically endangered. Although the species has been registered in several localities, its geographic distribution remains unclear, and the effects of climate change and current land uses on this species are largely unknown. We modeled the ecological niche of T. pinchaque using MaxEnt, in order to assess its potential adaptation to present and future climate change scenarios. We evaluated the effects of habitat loss due by current land use, the ecosystem availability and importance of Ecuadorian System of Protected Areas into the models. The model of environmental suitability estimated an extent of occurrence for species of 21,729 km2 in all of Ecuador, mainly occurring along the corridor of the eastern Ecuadorian Andes. A total of 10 Andean ecosystems encompassed ~98% of the area defined by the model, with herbaceous paramo, northeastern Andean montane evergreen forest and northeastern Andes upper montane evergreen forest being the most representative. When considering the effect of habitat loss, a significant reduction in model area (~17%) occurred, and the effect of climate change represented a net reduction up to 37.86%. However, the synergistic effect of both climate change and habitat loss, given current land use practices, could represent a greater risk in the short-term, leading to a net reduction of 19.90 to 44.65% in T. pinchaque's potential distribution. Even under such a scenarios, several Protected Areas harbor a portion (~36 to 48%) of the potential distribution defined by the models. However, the central and southern populations are highly threatened by habitat loss and climate change. Based on these results and due to the restricted home range of T. pinchaque, its preference for upland forests and paramos, and its small estimated population size in the Andes, we suggest to maintaining its current status as Critically Endangered in Ecuador.

A new synonym for Pristimantis luscombei (Duellman and Mendelson 1995) and the description of a new species of Pristimantis from the upper Amazon basin (Amphibia: Craugastoridae).

We consider Pristimantis achuar as junior synonym of P. luscombei, based on morphological and genetic evidence. Paratype specimens of P. luscombei are part of a new species, which lead to taxonomic confusion regarding the identity of P. luscombei. We describe and name this new species as Pristimantis miktos sp. nov. from Juyuintza, Pastaza province, eastern lowlands of Ecuador. Morphological diagnostic characters used to distinguish the new species from other brownish Amazonian Pristimantis are: (1) skin of dorsum shagreen with scattered tubercles or pustules; (2) tympanum prominent; (3) a thick X-shaped scapular dermal ridge in males; and (4) an orange iris in life. Pristimantis miktos is an inhabitant of the lowlands forests of the Pastaza and Napo drainages in eastern Ecuador and northern Loreto in Peru, reaching elevations of up to 350 m; P. luscombei is widely distributed in the upper Amazon Basin of Ecuador, northern Peru and extreme western Brazil, up to 1000 m. Phylogenetic analyses reveals that P. luscombei and the new species are not closest relatives, as also deduced from morphological evidence.

Novel data on the ecology of Cochranella mache (Anura: Centrolenidae) and the importance of protected areas for this critically endangered glassfrog in the neotropics.

We studied a population of the endangered glassfrog, Cochranella mache, at Bilsa Biological Station, northwestern Ecuador, from 2008 and 2009. We present information on annual abundance patterns, behavioral ecology, habitat use and a species distribution model performed with MaxEnt. We evaluate the importance of the National System of Protected Areas (SNAP) in Colombia and Ecuador, under scenarios of climate change and habitat loss. We predicted a restricted environmental suitability area from 48,509 Km(2) to 65,147 Km(2) along western Ecuador and adjacent Colombia; ∼ 8% of the potential distribution occurs within SNAP. We examined four aspects of C. mache ecology: (1) ecological data suggests a strong correlation between relative abundance and rainfall, with a high probability to observe frogs through rainy months (February-May); (2) habitat use and the species distribution model suggest that this canopy dweller is restricted to small streams and rivulets in primary and old secondary forest in evergreen lowland and piedmont forest of western Ecuador, with predictions of suitability areas in adjacent southern Colombia; (3) the SNAP of Colombia and Ecuador harbor a minimum portion of the predicted model of distribution (<10%); and (4) synergetic effects of habitat loss and climate change reduces in about 95% the suitability areas for this endangered frog along its distributional range in Protected Areas. The resulting model allows the recognition of areas to undertake conservation efforts and plan future field surveys, as well as forecasting regions with high probability of C. mache occurrence in western Ecuador and southern Colombia. Further research is required to assess population tendencies, habitat fragmentation and target survey zones to accelerate the discovery of unknown populations in unexplored areas with high probability of suitability. We recommend that Cochranella mache must be re-categorized as "Critically Endangered" species in national and global status, according with criteria and sub-criteria A4, B1ab(i,ii,iii,iv),E.

A new species of small tree frog, genus Dendropsophus (Anura: Hylidae) from the eastern Amazon lowlands of Ecuador.

We describe a new species of the hylid frog genus Dendropsophus from Amazonian Ecuador. The new species, Dendropsophus shiwiarum sp. nov., appears to be part of the D. microcephalus species group. Dendropsophus shiwiarum differs from its congeners by a combination of: (1) tympanic membrane non-differentiated and annulus evident only ventrally, (2) disc of Finger III and Toe IV with pointed tip, (3) dorsal surfaces of digital discs with a conical tubercle as result of projection of distal phalanx, and (4) small size (SVL < 19 mm). The new species is morphologically most similar to Dendropsophus riveroi. Examination of the type material of the latter suggests that some Ecuadorian populations reported as D. riveroi were misidentified and in fact belong to D. shiwiarum sp. nov.