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.

Taxonomic and Feeding Trait-Based Analysis of Macroinvertebrates in the Antisana River Basin (Ecuadorian Andean Region).

High-elevation tropical streams are under increasing threat from human activities and climate change. Specifically, Ecuadorian Andean streams require priority actions such as bioassessment (e.g., biodiversity and functional ecology of macroinvertebrates) in order to generate adequate environmental management policies. Therefore, we investigated the distribution and composition of the macroinvertebrate taxa and their functional feeding groups in relation to the environmental variables in the Antisana river basin (Andean-Ecuadorian Region). We sampled macroinvertebrates from 15 locations to assess ecological conditions (ECs), expressed as the Biological Monitoring Working Party Colombia (BMWP-Col) classes, the Andean Biotic Index (ABI) and the Andean-Amazon Biotic Index (AAMBI). Results indicate that dissolved oxygen saturation, elevation, nutrient concentration and conductivity contributed significantly to the composition of the taxa and functional feeding groups (FFGs). Taxa diversity and FFGs were more abundant in the best EC sites. Shredders (SH) were, overall, dominant and abundant at sites with medium-high ECs. Scrapers constituted the second most prevalent assemblage, exerting dominance at moderate ecological conditions (high altitude and high oxygen saturation). Collector-gathers (CGs) are less sensitive to contamination than the previous two groups but were equally abundant at medium-high EC sites. Collector-filterers (CFs) and parasites (PAs) were less abundant, although the presence of the former was slightly related to better environmental conditions. Predators (PRs) were almost absent throughout the study, but they were collected from poor EC sites. CGs, PAs and PRs showed more tolerance to the presence of human disturbances (e.g., hydraulic constructions or slope erosion). The BMWP-Col index seems to be the best fit for this ecosystem, showing a significant difference in FFG between the index classes, compared to the other indices evaluated. The results of this investigation may be regarded as a fundamental starting point and used in future bioassessment work in other similar ecosystems, particularly high-altitude tropical Ecuadorian streams.

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.

Polluted paradise: Occurrence of pesticide residues within the urban coastal zones of Santa Cruz and Isabela (Galapagos, Ecuador).

Although pesticides are frequently used for agriculture in the Galapagos Islands (Ecuador), there are, to date, no investigations of pesticide occurrences in its coastal waters. We examined the presence of pesticide residues in the coastal waters of urban areas in two islands of the Galapagos archipelago using a repeated sampling design. Quantification was performed by solid-phase extraction, followed by chemical analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-electron capture detector (GC-ECD). The diversity and concentration of pesticide residues in Santa Cruz island were higher compared to Isabela island. In total, sixteen pesticides were detected, including three persistent organic pollutants. Carbendazim (23.93 µg·L-1), cadusafos (4.74 µg·L-1), DDT (2.99 µg·L-1), diuron (1.61 µg·L-1) and aldrin (1.55 µg·L-1) were detected with the highest concentrations between samples. Repetitions in locations show that concentrations of pesticide residues varied considerably in space and time. Comparison with local products indicated agricultural activities on the islands as a possible source. Furthermore, evaluation through ecological risk quotients showed that the observed concentration levels of seven pesticides pose a relatively high risk for three biotic groups (i.e. algae, invertebrates and fishes). Taken together, this study provides insights into the need to regulate, monitor and assess the presence of pesticides in the islands. At a global scale, this study is moreover valuable for the many islands that are facing the same challenges.

First-generation teeth in nonmammalian lineages: evidence for a conserved ancestral character?

The present study focuses on the main characteristics of first-generation teeth (i.e., the first teeth of the dentition to develop in a given position and to become functional) in representatives of the major lineages of nonmammalian vertebrates (chondrichthyans, actinopterygians, and sarcopterygians: dipnoans, urodeles, squamates, and crocodiles). Comparative investigations on the LM and TEM level reveal the existence of two major types of first-generation teeth. One type (generalized Type 1) is characterized by its small size, conical shape, atubular dentine, and small pulp cavity without capillaries and blood vessels. This type is found in actinopterygians, dipnoans, and urodeles and coincides with the occurrence of short embryonic periods in these species. The other type assembles a variety of first-generation teeth, which have in common that they represent miniature versions of adult teeth. They are generally larger than the first type, have more complex shapes, tubular dentine, and a large pulp cavity containing blood vessels. These teeth are found in chondrichtyans, squamates, and crocodiles, taxa which all share an extended embryonic period. The presence in certain taxa of a particular type of first-generation teeth is neither linked to their phylogenetic relationships nor to adult body size or tooth structure, but relates to the duration of embryonic development. Given that the plesiomorphic state in vertebrates is a short embryonic development, we consider the generalized Type 1 first-generation tooth to represent an ancestral character for gnathostomes. We hypothesize that an extended embryonic development leads to the suppression of tooth generations in the development of dentition. These may still be present in the form of rudimentary germs in the embryonic period. In our view, this generalized Type 1 first-generation teeth has been conserved through evolution because it represents a very economic and efficient way of building small and simple teeth adapted to larval life. The highly adapted adult dentition characteristic for each lineage has been possible only through polyphyodonty.