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.

Recruitment and mortality of Rhizophora mangle L. seedlings in the Tropical Southwestern Atlantic mangrove.

Studies in the long-term recruitment and mortality of mangrove seedlings can help to understand mangrove demography and its relationship with climatic variables, environmental restoration and advances in the ecology of this ecosystem. A seven-year population dynamics study of seedling recruitment and mortality in cohorts of Rhizophora mangle L. was carried out to identify expansion processes and patterns of survival in the understory of mangrove forests on the Atlantic coast of Brazil. The present study aimed to evaluate the relationship between recruitment and mortality R. mangle seedlings at the population level, salinity, and climatic variables (precipitation, temperature and humidity). On an annual scale, seedling recruitment was positively correlated with mean temperature. Seedling density was negatively correlated with the number of recruits and positively with the number of deaths. The number of recruits was associated with dead seedlings, temperature and precipitation considering a population scale, without grouping the data. The seedling density in the stands increased with the number of dead seedlings. Our findings described the relationship between climate variability (durability and magnitude of the dry/rainy season) and the long-term population dynamics of R. mangle seedlings in a poorly studied region and from what moment, on a monthly and annual time scale, did this relationship become significant and changes occur. The findings of this study provide information on the population dynamics of the species that will help in understanding mangrove demography. These results have important implications for projections about the recruitment and survival of the species thinking about to long-term climate change that will modify current weather patterns and mangrove conservation efforts.

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.

Evaluation of the antinociceptive effect generated by citronellal monoterpene isomers.

Due to the complex nature of pain and the participation of physical, cognitive, psychological and behavioral aspects, pain management has several approaches. The use of medicinal plants in developing countries is quite expressive. Seeking new options for the treatment of emerging or debilitating diseases. Therefore, the present study seeks to elucidate the effects of the monoterpene, citronellal, differentiating its activity by isomers (R)-(+) and (S)-(-) citronellal. The study used several methods to evaluate the effects of citronellal isomers on motor coordination, nociceptive response, and the involvement of opioid, glutamatergic, and transient receptor pathways. The methods included rota-rod, hot-plate, and formalin tests, as well as the use of specific inhibitors and agonists. Data were analyzed using inferential statistics with a 95% confidence level. Both isomers did not significantly affect the motor coordination of the studied animals. The isomer (S)-(-) citronellal showed better results in relation to its structural counterpart, managing to have an antinociceptive effect in the formalin and hot plate tests with a lower concentration (100 mg/kg) and presenting fewer side effects, however, the this study was not able to elucidate the mechanism of action of this isomer despite having activity in studies with substances that act on specific targets such as glutamate and capsaicin, its activity was not reversed with the use of antagonists for pathways related to nociception. While the (R)-(+) citronellal isomer, despite showing total activity only at a concentration of 150 mg/kg, was able to determine its mechanism of action related to the opioid pathway by reversing its activity by the antagonist naloxone, being this is a pathway already correlated with nociception control treatments, however, it is also related to some unwanted side effects. In this way, new studies are sought to elucidate the mechanism related to the isomer (S)-(-) citronellal and a possibility of use in other areas related to the treatment of pain or inflammation.

Características clínicas y resultados de pacientes con fibrosis quística con enfermedad pulmonar avanzada: experiencia en 10 años del Instituto Nacional del Tórax / Clinical features and outcomes of cystic fibrosis patients with advanced lung disease: experience of 10 years at the Instituto Nacional del Tórax

La enfermedad pulmonar avanzada (EPAV) es la principal causa de morbimortalidad en pacientes con Fibrosis Quística (FQ). Objetivo: describir características clínicas de pacientes con FQ con EPAV y mortalidad en el seguimiento. Método: Estudio descriptivo, retrospectivo de pacientes con FQ y EPAV: VEF1 4 años de vida. Un 75% era portador de infección crónica por Pseudomonas. Un 68% era dependiente de oxígeno y un 18% de ventilación mecánica no invasiva. El 70 % tuvo 2 o más hospitalizaciones el último año de seguimiento. De 27 pacientes derivados a trasplante, 7 se trasplantaron, 3 fallecieron en lista para trasplante, 9 presentaron alguna contraindicación: 4 de ellos por desnutrición y 5 por mala adherencia y escasa red de apoyo. En el seguimiento un 32% (n = 14) falleció, 93% de causa respiratoria. Conclusión: Un 39% de los pacientes tenían EPAV cuyo diagnóstico de FQ en promedio fue a los 11,2 años (SD ± 13 a). Las barreras de ingreso a lista para trasplante fueron: desnutrición, mala adherencia y falta de red de apoyo. Esta es una población con una elevada mortalidad.

Advanced cystic fibrosis lung disease (ACFLD) is the leading cause of morbidity and mortality in patients with Cystic Fibrosis (CF). Objective: to describe clinical characteristics of patients with CF with ACFLD and mortality during follow-up. Method: Descriptive, retrospective study of patients with CF and ACFLD: FEVi < 40%, oxygen dependent, and/or referred to a lung transplantprogram. Clinical, microbiological, functional, genetic and mortality characteristics were collected. Results: Of 111 controlled patients, 39% met criteria for ACFLD. 52% were men and the mean age was 29,8 years- old. The average BMI was 19.9 kg/m2, 72% had pancreatic insufficiency and 87% had a genetic study, being the DF508 mutation the most frequent (67%). The average age of diagnosis was 11.2 years (SD ± 13 years), being in 54,5% over the age of 4 years. 75% had chronic Pseudomonas infection. 68% were oxygen dependent and 18% on noninvasive mechanical ventilation. In the last year of follow-up 70% had 2 or more hospitalizations. Of 27 patients who have been referred for transplantation, 7 underwent lung transplantation, 3 died waiting on the transplant list, 9 had contraindications: 4 due to malnutrition and 5 to poor adherence and poor support network. 32% (n = 14) of the ACFLD patients died, 93% due to respiratory causes. Conclusion: 39% of the patients had ACFLD. The average age for CF diagnosis was 11.2 years (SD ± 13 years) Barriers to entering the transplant list are: malnutrition, poor adherence, and lack of a support network. This is a population with a high mortality.

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.

Effects of Microabrasion Prior to In-office Bleaching on Hydrogen Peroxide Permeability, Color Change, and Enamel Morphology.

PURPOSE: This study evaluated hydrogen peroxide (HP) diffusion within the pulp chamber, as well as color change and the surface morphology of teeth subjected to various microabrasion (MA) protocols associated or not with in-office (IO) bleaching. METHODS: Forty sound premolars were randomly divided into the following four groups (n=10): no treatment (NC); IO bleaching only; IO immediately after MA (IMA), and IO seven days after MA (7MA). After treatments, the HP concentration (µg/mL) within the pulp chamber was determined using ultraviolet-visible (UV-Vis) spectrophotometry. The color change (ΔE*) was evaluated using the digital spectrophotometer before and 1 week after bleaching. The surface morphology was evaluated by scanning electron microscope (SEM). Data from each test were submitted to one-way ANOVA and Tukey tests (α=0.05). RESULTS: All experimental groups exhibited higher HP concentrations compared to the NC group (p<0.00001). However, higher amounts of HP were observed for the IMA group compared to the IO and 7MA groups (p<0.00001). No significant difference in color change was observed among the groups (p<0.001). Pronounced grooves in enamel were found in the IMA and 7MA groups. However, enamel erosion areas were observed only in the 7MA group. CONCLUSIONS: The association between MA and IO bleaching could significantly affect the amount of HP inside the pulp chamber. Therefore, it is highly recommended to wait for 1 week after MA procedures before performing IO bleaching.

Review of global neurosurgery education: Horizon of Neurosurgery in the Developing Countries.

Globally, the discipline of neurosurgery has evolved remarkably fast. Despite being one of the latest medical specialties, which appeared only around hundred years ago, it has witnessed innovations in the aspects of diagnostics methods, macro and micro surgical techniques, and treatment modalities. Unfortunately, this development is not evenly distributed between developed and developing countries. The same is the case with neurosurgical education and training, which developed from only traditional apprentice programs in the past to more structured, competence-based programs with various teaching methods being utilized, in recent times. A similar gap can be observed between developed and developing counties when it comes to neurosurgical education. Fortunately, most of the scholars working in this field do understand the coherent relationship between neurosurgical education and neurosurgical practice. In context to this understanding, a symposium was organized during the World Federation of Neurological Surgeons (WFNS) Special World Congress Beijing 2019. This symposium was the brain child of Prof. Yoko Kato-one of the eminent leaders in neurosurgery and an inspiration for female neurosurgeons. Invited speakers from different continents presented the stages of development of neurosurgical education in their respective countries. This paper summarizes the outcome of these presentations, with particular emphasis on and the challenges faced by developing countries in terms of neurosurgical education and strategies to cope with these challenges.

Short- and medium-term prognosis of HIV-infected patients receiving intensive care: a Brazilian multicentre prospective cohort study.

OBJECTIVES: The characteristics of critically ill HIV-positive patients and the causes of their admission to intensive care units (ICUs) are only known through retrospective and unicentric studies. This study aims to fill this knowledge gap. METHODS: This is a prospective, multicentre cohort study of short- and medium-term prognostic factors. The setting consisted of ICUs of three tertiary referral hospitals from the three largest metropolitan areas in Brazil in the period January 2014 to November 2015. In all, 161 HIV patients over 18 years old were included. RESULTS: The clinical data of the outcomes (ICU mortality, hospital mortality and 90-day survival) were extracted from medical records using the REDCap®ï¸ web-based form and analysed with the MedCalc®ï¸ application. Median age was 41.7 [interquartile range (IQR): 34-50] years, the Simplified Acute Physiologic Score 3 (SAPS 3) was 64 (IQR: 56-74), and the Sequential Organ Failure Assessment Score (SOFA) was 6 (IQR: 4-9) points. The main causes of admission were sepsis (54.5%) and acute respiratory failure (13.7%). ICU and hospital mortality rates were 32.3% and 40.4%, respectively. In a multivariate analysis, time until ICU admission ≥ 3 days (P = 0.0013), performance status (Eastern Cooperative Oncology Group score, P = 0.0344), coma (Glasgow Coma Scale ≤ 8 points, P = 0.0213) and sepsis (P = 0.0003) were associated with increased hospital mortality. Coma (P = 0.0002) and sepsis (P = 0.0008) were independently associated with 90-day survival. CONCLUSIONS: Delayed ICU admission and the severity of critical illness determine the short- and medium-term mortality rates of HIV-infected patients admitted to the ICU, rather than factors associated with HIV infection. These results suggest that prognostic factors of HIV-infected patients in the ICU are similar to those of non-HIV-infected populations.

Correction to: Review of global neurosurgery education: Horizon of Neurosurgery in the Developing Countries.

[This corrects the article DOI: 10.1186/s41016-020-00194-1.].

A large familial cluster and sporadic cases of frontal fibrosing alopecia in Brazil reinforce known human leucocyte antigen (HLA) associations and indicate new HLA susceptibility haplotypes.

BACKGROUND: Frontal fibrosing alopecia (FFA) is a lymphocytic scarring alopecia whose worldwide incidence is rising. Environmental triggers combined with genetic predisposition represent one of the current hypotheses in FFA aetiology. Familial clusters are opportunities to investigate the genetic basis of diseases. OBJECTIVES: Assess human leucocyte antigen (HLA) genetic variability in a Brazilian sample of a large familial cluster (six sisters and one daughter) with FFA, unnafected familiar members and sporadic cases of FFA. METHODS: We addressed the HLA-A, HLA-B, HLA-C, HLA-G and HLA-E genetic variability in this family and in seven sporadic FFA cases, comparing allele frequencies with those reported for the São Paulo State from Brazil. RESULTS: Two susceptibility haplotypes, C*17:01:01:02/B*42:01:01:01 and C*07:02:01:03/B*07:02:01:01, were identified among familial cases and also in sporadic cases. The first haplotype is rare among Brazilians, and it was not previously reported as being associated with FFA. Both alleles were found in some different unaffected familiars, what emphasizes the role of environmental triggers in disease development. HLA-A, HLA-G and HLA-E genes were not associated to familiar nor FFA sporadic cases. CONCLUSION: The identification of susceptibility haplotypes in FFA reinforces the genetic predisposition to the disease.

Comparison of antibody repertories against Staphylococcus aureus in healthy and infected dairy cows with a distinct mastitis history and vaccinated with a polyvalent mastitis vaccine.

Staphylococcus aureus is one of the pathogens most frequently isolated from cases of mastitis worldwide. To decrease the effect of S. aureus mastitis in dairy farming, alternative strategies for controlling mastitis are needed that depend on a better knowledge of cow-to-cow variations in S. aureus antibody production. The present study sought to explore the diversity of S. aureus antibodies produced by dairy cows with a distinct mastitis history and vaccinated with a polyvalent mastitis vaccine. We obtained protein extracts from S. aureus isolates derived from persistent subclinical mastitis. Proteins were fractionated using 2-dimensional gel electrophoresis and Western blotting. Then, Western blotting membranes were exposed to sera from 24 dairy cows that had been divided into the following groups: vaccinated dairy cows that were infected with S. aureus, further subdivided according to whether they (a) remained infected by S. aureus or (b) recovered from the intramammary infection; unvaccinated dairy cows infected with S. aureus; and vaccinated healthy dairy cows with no history of S. aureus mastitis. Proteins found to be reactive by Western blot were identified by mass spectrometry (MALDI/TOF-TOF). Our most important finding was that F0F1 ATP synthase subunit α, succinyl-diaminopimelate desuccinylase, and cysteinyl-tRNA synthetase were potential candidate proteins for the prevention of S. aureus mastitis. This study strengthens the notion that variations among animals should not be ignored and shows that the heterogeneity of antibody production against anti-staphylococcal antigens in animals may enable the identification of new immunotherapy targets.

Records of breeding in Wilson's Plover Charadrius wilsonia with new localities for Brazil / Registros da reprodução de Charadrius wilsonia, com novas ocorrências para o Brasil

Abstract Wilson's Plover, Charadrius wilsonia, is widely distributed in coastal areas of the Americas. This report presents the first record of breeding in this species on Coroa do Avião Island, on the coast of Pernambuco, and in the estuary of the Cardoso and Camurupim rivers, on the coast of Piauí, in northeastern Brazil, extending the known area of reproduction of this species in this region. One breeding pair was observed on October 29th 2014 on Coroa do Avião Island, and a second pair was recorded on April 6th 2016 in the Cardoso/Camurupim estuary on the Piauí coast. Both the male and the female contributed to the incubation of the eggs. The nest on Coroa do Avião island was camouflaged by the local vegetation, but despite this, the eggs were attacked by a predator. Possible predators observed on the island included Caracara plancus and domestic cats and dogs.

Resumo Charadrius wilsonia (Wilson's Plover) está amplamente distribuído pela costa das Américas. Este é o primeiro registro de reprodução no litoral de Pernambuco, Coroa do Avião, e no estuário dos rios Cardoso e Camurupim, litoral do Piauí, Brasil, ampliando a área de reprodução no Nordeste do Brasil. Foi encontrado um casal em período reprodutivo em 23 de outubro de 2014 na ilha Coroa do Avião, litoral de Pernambuco e outro registro em 06 de abril de 2016 nos estuários Cardoso e Camurupim, litoral do Piauí. Foi observado que o macho e a fêmea contribuem na incubação dos ovos. A vegetação na ilha contribui para a camuflagem do ninho, bem como na proteção dos ovos pelos predadores. Apesar da proteção ocorreu a predação do ninho na ilha Coroa do Avião. Alguns possíveis predadores foram registrados na ilha, como Caracara plancus que diariamente frequentam a área e animais domésticos como cães e gatos.

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.

Records of breeding in Wilson's Plover Charadrius wilsonia with new localities for Brazil.

Wilson's Plover, Charadrius wilsonia, is widely distributed in coastal areas of the Americas. This report presents the first record of breeding in this species on Coroa do Avião Island, on the coast of Pernambuco, and in the estuary of the Cardoso and Camurupim rivers, on the coast of Piauí, in northeastern Brazil, extending the known area of reproduction of this species in this region. One breeding pair was observed on October 29th 2014 on Coroa do Avião Island, and a second pair was recorded on April 6th 2016 in the Cardoso/Camurupim estuary on the Piauí coast. Both the male and the female contributed to the incubation of the eggs. The nest on Coroa do Avião island was camouflaged by the local vegetation, but despite this, the eggs were attacked by a predator. Possible predators observed on the island included Caracara plancus and domestic cats and dogs.

Chemiluminescent ELISA with multi-epitope proteins to improve the diagnosis of canine visceral leishmaniasis.

Diagnosing canine visceral leishmaniasis (CVL) is difficult because clinical signs of the disease are non-specific and a many infected animals in endemic areas, as in Brazil, are asymptomatic. Serological tests are the most common diagnostic methods employed, but most have limitations. For this reason, the implementation of a rapid, sensitive, and specific diagnostic test for CVL has become increasingly important. In this study, we adapted a chemiluminescent enzyme-linked immunosorbent assay (CL ELISA), using two multi-epitope recombinant proteins (PQ10 and PQ20) and a crude Leishmania antigen produced using promastigotes of L. infantum, as antigens to detect CVL infection in animals from Belo Horizonte. To investigate cross-reactions, samples from dogs with other infections (babesiosis, ehrlichiosis and Trypanosoma cruzi) were tested. Assay performance validations were conducted to analyse parameters such as variability, reproducibility, and stability. CL ELISA sensitivity/specificity with PQ10 antigen was 93.1%/80.0%; with the PQ20 protein 93.1%/96.6%; and with the crude antigen 75%/73.3%. Inter-assay variability and inter-operator coefficient of variation were <7% and <15%, with PQ10 and PQ20, respectively. The accuracy of the CL ELISA was classified as excellent for PQ10 (AUC = 0.95) and PQ20 (AUC = 0.98) and moderate for the crude antigen (AUC = 0.77). The kappa score for qualitative agreement between two plate lots was excellent for PQ10 (0.89) and good for PQ20 (0.65). PQ20 remained more stable than PQ10. The CL ELISA with recombinant proteins is a promising tool to diagnose CVL.

Cardiac protein expression patterns are associated with distinct inborn exercise capacity in non-selectively bred rats.

In the present study, we successfully demonstrated for the first time the existence of cardiac proteomic differences between non-selectively bred rats with distinct intrinsic exercise capacities. A proteomic approach based on two-dimensional gel electrophoresis coupled to mass spectrometry was used to study the left ventricle (LV) tissue proteome of rats with distinct intrinsic exercise capacity. Low running performance (LRP) and high running performance (HRP) rats were categorized by a treadmill exercise test, according to distance run to exhaustion. The running capacity of HRPs was 3.5-fold greater than LRPs. Protein profiling revealed 29 differences between HRP and LRP rats (15 proteins were identified). We detected alterations in components involved in metabolism, antioxidant and stress response, microfibrillar and cytoskeletal proteins. Contractile proteins were upregulated in the LVs of HRP rats (α-myosin heavy chain-6, myosin light chain-1 and creatine kinase), whereas the LVs of LRP rats exhibited upregulation in proteins associated with stress response (aldehyde dehydrogenase 2, α-crystallin B chain and HSPß-2). In addition, the cytoskeletal proteins desmin and α-actin were upregulated in LRPs. Taken together, our results suggest that the increased contractile protein levels in HRP rats partly accounted for their improved exercise capacity, and that proteins considered risk factors to the development of cardiovascular disease were expressed in higher amounts in LRP animals.

A proteomic road to acquire an accurate serological diagnosis for human tegumentary leishmaniasis.

Diagnostic tools are important for clinical management and epidemiological evaluation of Tegumentary (TL) and Visceral (VL) Leishmaniasis. Serology is not frequently used for the diagnosis of the TL form because low antibody titers and cross-reaction with VL. Therefore, it is crucial to identify specific and immunogenic antigens from species associated with the TL form. Here we employed a proteomic approach coupled to an in silico analysis and identified the most abundant and immunogenic proteins from Leishmania amazonensis, Leishmania braziliensis and Leishmania infantum. Of 16 species specific proteins, nine were from the species causative of the TL form (L. amazonensis and L. braziliensis). In silico analysis revealed 18 B-cell epitopes with 0% similarity to Trypanosoma cruzi orthologs and, therefore, less likely to crossreact with sera of patients with Chagas disease. Two proteins reacted exclusively with serum from TL patients and presented several B-cell epitopes without similarity to T. cruzi orthologs: the hypothetical protein GI 134063939 and the metallo-peptidase Clan MA(E)-Family M3. The immunoassay revealed nine peptides with strong reactivity to sera from TL patients. These proteins and peptides may be good candidates to improve the specificity and sensibility of serological tests aiming to diagnose the TL of this neglected human disease. BIOLOGICAL SIGNIFICANCE: As no gold-standard test for tegumentary leishmaniasis (TL) exists, a combination of different diagnostic techniques is often necessary to obtain precise results. Thus, the identification of species-specific, highly immunogenic and abundant proteins that stimulate the humoral immune response in the host should help in the development of serological tests for human TL. Herein we searched for these potential antigens in Leishmania species related to American Leishmaniasis (L. amazonensis, L. braziliensis and L. infantum). To this end, we employed an immunoproteomic approach using proteins from these Leishmania species and sera from TL and Visceral Leishmaniasis (VL) patients. Our study unveils specific proteins and peptides that may represent antigens that will help the efforts to improve the accuracy of serological tests to diagnose the TL form.