Perspectives on label-free microscopy of heterogeneous and dynamic biological systems.

Significance: Advancements in label-free microscopy could provide real-time, non-invasive imaging with unique sources of contrast and automated standardized analysis to characterize heterogeneous and dynamic biological processes. These tools would overcome challenges with widely used methods that are destructive (e.g., histology, flow cytometry) or lack cellular resolution (e.g., plate-based assays, whole animal bioluminescence imaging). Aim: This perspective aims to (1) justify the need for label-free microscopy to track heterogeneous cellular functions over time and space within unperturbed systems and (2) recommend improvements regarding instrumentation, image analysis, and image interpretation to address these needs. Approach: Three key research areas (cancer research, autoimmune disease, and tissue and cell engineering) are considered to support the need for label-free microscopy to characterize heterogeneity and dynamics within biological systems. Based on the strengths (e.g., multiple sources of molecular contrast, non-invasive monitoring) and weaknesses (e.g., imaging depth, image interpretation) of several label-free microscopy modalities, improvements for future imaging systems are recommended. Conclusion: Improvements in instrumentation including strategies that increase resolution and imaging speed, standardization and centralization of image analysis tools, and robust data validation and interpretation will expand the applications of label-free microscopy to study heterogeneous and dynamic biological systems.

4-Nitrophenol at environmentally relevant concentrations mediates reproductive toxicity in Caenorhabditis elegans via metabolic disorders-induced estrogen signaling pathway.

4-Nitrophenol (4-NP), as a toxic and refractory pollutant, has generated significant concern due to its adverse effects. However, the potential toxic effects and mechanism remained unclear. In this study, the reproduction, development, locomotion and reactive oxygen species (ROS) production of Caenorhabditis elegans were investigated to evaluate the 4-NP toxicity. We used metabolomics to assess the potential damage mechanisms. The role of metabolites in mediating the relationship between 4-NP and phenotypes was examined by correlation and mediation analysis. 4-NP (8 ng/L and 8 µg/L) caused significant reduction of brood size, ovulation rate, total germ cells numbers, head thrashes and body bends, and an increase in ROS. However, the oosperm numbers in uterus, body length and body width were decreased in 8 µg/L. Moreover, 36 differential metabolites were enriched in the significant metabolic pathways, including lysine biosynthesis, ß-alanine metabolism, tryptophan metabolism, pentose phosphate pathway, pentose and glucuronate interconversions, amino sugar and nucleotide sugar metabolism, starch and sucrose metabolism, galactose metabolism, propanoate metabolism, glycerolipid metabolism, and estrogen signaling pathway. The mechanism of 4-NP toxicity was that oxidative stress caused by the perturbation of amino acid, which had effects on energy metabolism through disturbing carbohydrate and lipid metabolism, and finally affected the estrogen signaling pathway to exert toxic effects. Moreover, correlation and mediation analysis showed glycerol-3P, glucosamine-6P, glucosamine-1P, UDP-galactose, L-aspartic acid, and uracil were potential markers for the reproduction and glucose-1,6P2 for developmental toxicity. The results provided insight into the pathways involved in the toxic effects caused by 4-NP and developed potential biomarkers to evaluate 4-NP toxicity.

Impact of residual antibiotics on microbial decomposition of livestock manures in Eutric Regosol: Implications for sustainable nutrient recycling and soil carbon sequestration.

The land application of livestock manure has been widely acknowledged as a beneficial approach for nutrient recycling and environmental protection. However, the impact of residual antibiotics, a common contaminant of manure, on the degradation of organic compounds and nutrient release in Eutric Regosol is not well understood. Here, we studied, how oxytetracycline (OTC) and ciprofloxacin (CIP) affect the decomposition, microbial community structure, extracellular enzyme activities and nutrient release from cattle and pig manure using litterbag incubation experiments. Results showed that OTC and CIP greatly inhibited livestock manure decomposition, causing a decreased rate of carbon (28%-87%), nitrogen (15%-44%) and phosphorus (26%-43%) release. The relative abundance of gram-negative (G-) bacteria was reduced by 4.0%-13% while fungi increased by 7.0%-71% during a 28-day incubation period. Co-occurrence network analysis showed that antibiotic exposure disrupted microbial interactions, particularly among G- bacteria, G+ bacteria, and actinomycetes. These changes in microbial community structure and function resulted in decreased activity of urease, ß-1,4-N-acetyl-glucosaminidase, alkaline protease, chitinase, and catalase, causing reduced decomposition and nutrient release in cattle and pig manures. These findings advance our understanding of decomposition and nutrient recycling from manure-contaminated antibiotics, which will help facilitate sustainable agricultural production and soil carbon sequestration.

Variación espacio-temporal de la diversidad de escarabajos Geadephaga (Coleoptera: Carabidae, Cicindelidae) en el bosque seco tropical del Caribe colombiano / Spatio-temporal variation in the diversity of Geadephaga beetles (Coleoptera: Carabidae, Cicindelidae), in the tropical dry forest of the Colombian Caribbean

Resumen Introducción: El uso de artrópodos en estudios ecológicos en el bosque seco tropical (Bs-T) de Colombia se centra mayormente en arañas, mariposas, hormigas y escarabajos coprófagos; por lo tanto, es necesario comprender como es la dinámica estacional de otros grupos como Geadephaga en este ecosistema. Objetivo: Evaluar la variación espacio-temporal y el efecto de las condiciones ambientales sobre la diversidad de las comunidades de Geadephaga en dos fragmentos de Bs-T en el Caribe colombiano. Métodos: El estudio se llevó a cabo en cuatro eventos de muestreo entre febrero y junio de 2018; cada uno con una duración de cuatro días y cuatro noches. En cada localidad (Reserva Campesina La Montaña = RCM y Reserva La Flecha = RLF), se marcaron cuatro estaciones distanciadas 350 m, con parcelas de 50×50 m. En el centro de cada parcela se instaló una trampa de luz, mientras que en los cuatro vértices se instalaron trampas de caída, y se realizó captura manual, cernido de hojarasca y perturbación de follaje. Resultados: Se capturó un total de 348 ejemplares de Carabidae y 114 de Cicindelidae. La riqueza y la abundancia presentaron los valores más bajos durante el período seco, mientras que los mayores valores se observaron durante el período de lluvias, en ambos fragmentos. Tetracha affinis (Dejean, 1825) fue la especie más abundante en la RCM durante el período lluvioso. Conclusiones: La variación de la riqueza y abundancia de Geadephaga responde a un patrón estacional en ambos fragmentos. Se encontró una amplia disimilitud entre las comunidades de Geadephaga de ambos fragmentos, pese a tener condiciones ambientales parecidas.

Abstract Introduction: The use of arthropods in ecological studies in the tropical dry forest (TDF) of Colombia focused mainly on spiders, butterflies, ants, and dung beetles; therefore, it is necessary to understand the seasonal dynamics of the other groups such as Geadephaga in this ecosystem. Objective: To evaluate the spatio-temporal variations and the effect of environmental conditions on the diversity of the Geadephaga communities in two TDF fragments in the Colombian Caribbean. Methods: This study was conducted during four sampling events between February and June 2018, each lasting four days, and four nights. Four sampling stations were selected in each locality (Reserva Campesina La Montaña = RCM and Reserva La Flecha = RLF), separated 350 m one from another, with square plots of 50 × 50 m. A light trap was installed in the center of each square, while pitfall traps were installed in the four vertices, and manual capture, beating sheets and leaf litter sieve were carried out. Results: A total of 348 Carabidae and 114 of Cicindelidae were captured. In both fragments, the richness and abundance presented the lowest values in dry season, while the highest values were observed during the rainy season. Tetracha affinis (Dejean, 1825) was the most abundant species in RCM during rainy season. Conclusions: The richness variation and abundance of Geadephaga corresponds to a seasonal pattern at both locations. A high dissimilarity between the communities of Geadephaga from the two fragments was found, despite having similar environmental conditions.

Spatio-temporal composition of aquatic birds communityin Juluapan Lagoon, Colima, Mexican Central Pacific / Composición espacio-temporal de las aves acuáticas en la laguna Juluapan, Colima, en el Pacífico Central Mexicano

Abstract Introduction: Aquatic birds (AB) are usually associated with wetlands, which provide refuge, food, and/or nesting sites for resident and migratory species. Despite their ecological importance, there is little knowledge on AB in some tropical environments, such as those found on the Colima coast. Objective: To investigate the spatial and temporal composition of the AB community in Juluapan Lagoon, Colima, Central Mexican Pacific. Methods: Monthly counts were conducted between June 2017 and May 2018 during low-tide conditions to record habitat use by AB. Species richness and bird counts were obtained to compare sampling areas; mean richness and number of individuals were compared between seasons. Results: We detected 53 species and 5 750 individuals. The highest species richness and relative abundance values were obtained in winter at the lagoon area farthest from the connection with the marine system, where anthropogenic activity is lower. Diversity was greater in zones 2 and 3 in spring, summer, and fall. Muddy flats were the most used environment, and the most frequent activity was resting. Nesting activity was only recorded in the middle of the lagoon at the mangrove during spring. "Shorebirds" and "waders" were the most dominant groups in the bird community of the Juluapan lagoon. Conclusions: This coastal wetland is a site of great biological importance for aquatic birds; thus, conservation measures should be implemented, and there should be a continuous study of the effects of anthropogenic pressure.

Resumen Introducción: Las aves acuáticas (AA) son usualmente relacionadas a los humedales debido a que éstos funcionan como sitios de refugio, alimentación y anidación de diferentes especies residentes y migratorias. Sin embargo, el conocimiento sobre las aves acuáticas en algunos humedales es nulo. Objetivo: Investigar la composición espacio-temporal de la comunidad de AA en la laguna Juluapan, Colima, en el Pacífico Central Mexicano. Métodos: Entre junio de 2017 y mayo de 2018 se llevaron a cabo conteos mensuales en condiciones de marea baja para registrar el uso de hábitat de las AA. Se obtuvieron valores de riqueza de especies y número de individuos para realizar comparaciones entre zonas de muestreo, así como el promedio del número de especies y número de individuos para comparaciones entre temporadas. Resultados: Se registraron un total de 53 especies y 5 750 individuos. Los valores de riqueza de especies y densidad de individuos fueron más altos durante invierno, en la zona más alejada al ambiente marino, donde la actividad antropogénica es menor. La diversidad tuvo valores más altos en la zona 2 y 3, durante primavera, verano y otoño. El ambiente más explotado por las aves fueron las planicies lodosas; y el descanso fue la actividad más frecuente. Asimismo, la actividad de anidación sólo se registró en el manglar de la zona media durante primavera. Las "aves playeras" y "aves zancudas" fueron los grupos más predominantes en la comunidad de aves de la laguna Juluapan. Conclusiones: Este humedal costero es un sitio de gran importancia biológica para aves acuáticas, por lo que resulta necesario la implementación de medidas de conservación, así como el estudio de los efectos por la presión antropogénica.

Modelos de distribución de especies y estado de conservación de murciélagos amenazados en la región tumbesina de Ecuador y Perú / Species distribution models and conservation status of threatened bats in the Tumbesian region of Ecuador and Perú

Resumen Introducción: La biodiversidad se está perdiendo a un ritmo acelerado como resultado del cambio global. Herramientas como los modelos de distribución de especies (MDEs) han sido ampliamente usados para mejorar el conocimiento sobre el estado de conservación de las especies y ayudar a desarrollar estrategias de gestión para mitigar la pérdida de biodiversidad. Objetivo: Determinar cómo la distribución potencial predicha por los MDEs para ocho especies de murciélagos amenazados difiere de los mapas de distribución reportados por la UICN. También, inferir el área de distribución y estado de endemismo de cada especie, y evaluar la importancia de la región tumbesina para su conservación. Métodos: Basados en registros de presencia del rango global de las especies, usamos MDEs para evaluar el estado de conservación de estas ocho especies en la región tumbesina de Ecuador y Perú. Resultados: Las áreas estimadas por los MDEs eran 35-78 % más pequeñas para cuatro especies (Eptesicus innoxius, Lophostoma occidentale, Platalina genovensium y Lonchophylla hesperia) y 26-1 600 % más grandes para tres especies (Amorphochilus schnablii, Promops davisoni y Rhogeessa velilla) que aquellas reportadas por la UICN. Para Tomopeas ravus, el área estimada por el MDE y la UICN fue similar, pero difirió en la distribución espacial. Los MDEs coincidieron con áreas de endemismo informadas por autores previos para E. innoxius, R. velilla y T. ravus, pero fueron diferentes para A. schnablii, P. genovensium, P. davisoni y L. hesperia, debido en parte a las distribuciones proyectadas para estas últimas especies en valles secos interandinos según los MDEs. Conclusiones: La región tumbesina representa una porción significativa (40-96 %) de la distribución predicha de siete de las ocho especies estudiadas, subrayando la importancia de esta región para la conservación de murciélagos. Nuestros resultados muestran las probables distribuciones para estas especies y proporcionan una base importante para identificar vacíos de investigación y desarrollar medidas de conservación para murciélagos amenazados en el punto caliente de biodiversidad de Tumbes.

Abstract Introduction: Biodiversity is being lost at an accelerating rate because of global change. Tools such as species distribution models (SDMs) have been widely used to improve knowledge about species' conservation status and help develop management strategies to mitigate biodiversity loss. SDMs are especially important for species with restricted distributions, such as endemic species. Objective: To determine how potential distribution predicted by SDMs for eight threatened bat species differed from the distribution maps reported by the IUCN. Also, to infer the area of distribution and state of endemism of each specie, and to evaluate the importance of the Tumbesian region for their conservation. Methods: Based on presence records across the species' entire ranges, we used SDMs to assess the conservation status of these eight species in the Tumbesian region of Ecuador and Peru. Results: The areas estimated by SDMs were 35-78 % smaller for four species (Eptesicus innoxius, Lophostoma occidentale, Platalina genovensium and Lonchophylla hesperia) and 26-1 600 % larger for three species (Amorphochilus schnablii, Promops davisoni and Rhogeessa velilla) than those reported by the IUCN. For Tomopeas ravus, the area estimated by the SDM and IUCN was similar but differed in spatial distribution. SDMs coincided with areas of endemism reported by previous authors for E. innoxius, R. velilla, and T. ravus, but were different for A. schnablii, P. genovensium, P. davisoni, and L. hesperia, due in part to projected distributions for these latter species in dry inter-Andean valleys according to the SDMs. Conclusions: The Tumbesian region represents a significant portion (40-96 %) of the predicted distribution of seven of the eight species studied, underscoring the importance of this region for bat conservation. Our results show likely distributions for these species and provide an important basis for identifying research gaps and developing conservation measures for threatened bats in the Tumbes biodiversity hotspot.

The role of neophobia in a pioneer olfactory enrichment for amazons, macaws, and toucans / El papel de la neofobia en un enriquecimiento olfativo pionero para guacamayos, loros y tucanes

Abstract Introduction: Despite sensory enrichment being critical for ensuring the well-being of captive wild animals, smells are not being included in enrichment protocols for birds. For this group, neophobia can be a problem when it comes to implementing new enrichment devices. Objective: To explore how participation in an olfactory enrichment and latency times varies between bird taxonomic groups (Amazona spp. / Ara spp. / Ramphastos spp.). Methods: We exposed 257 birds to a scent enrichment, and we recorded which individuals engaged with it and the time they took to interact with it. Results: We discovered that participation by toucans in the enrichment was higher compared to amazons and macaws. Furthermore, latency time to interact with the enrichment was higher in amazons that in the other species. Our findings could suggest that toucans are neophilic species which could benefit from higher exploration rates. Amazons on the contrary seem to be particularly neophobic, possibly because of their less opportunistic feeding habits compared to toucans and their higher vulnerability to predation compared to macaws. Conclusion: These results point out that toucans would be more inclined to engage in environmental enrichments, while a more natural design using smells inside familiar objects could be a more successful enrichment for psittacids.

Resumen Introducción: A pesar de que el enriquecimiento sensorial es fundamental para garantizar el bienestar de los animales silvestres en cautiverio, los olores no son incluidos de forma rutinaria en los protocolos de enriquecimiento para aves. Además, en el caso de estos animales, la neofobia puede ser un problema a la hora de implementar nuevos dispositivos de enriquecimiento. Objetivo: Explorar cómo varía la participación y la latencia en la interacción con un enriquecimiento olfativo entre grupos taxonómicos de aves (Amazona spp. / Ara spp. / Ramphastos spp.). Métodos: Expusimos a 257 aves a un enriquecimiento olfativo y registramos qué individuos participaron y el tiempo que tardaron en interactuar con él. Resultados: La participación en el enriquecimiento fue mayor en los tucanes en comparación con las amazonas y los guacamayos. Además, el tiempo de latencia para interactuar con el enriquecimiento fue mayor en las amazonas que en las otras especies. Nuestros hallazgos sugieren que los tucanes son especies neofílicas que podrían beneficiarse de tasas de exploración más altas. Por otro lado, las amazonas parecen ser particularmente neofóbicas, posiblemente debido a sus hábitos alimenticios menos oportunistas en comparación con los tucanes y a su mayor vulnerabilidad a la depredación en comparación con los guacamayos. Conclusión: Estos resultados señalan que los tucanes serían más proclives a participar en enriquecimientos ambientales, mientras que un diseño que utilice olores dentro de objetos más naturales o familiares podría ser más exitoso para las psitácidas.

Environmental sympatry through time: spatio-temporal distribution and conservation status of two sympatric anuran species (Leptodactylidae) in South America / Simpatría ambiental a través del tiempo: distribución espacio-temporal y estado de conservación de dos especies de anuros simpátricos (Leptodactylidae) en América del Sur

Abstract Introduction: Leptodactylus latinasus and Physalaemus cuqui are sympatric anuran species with similar environmental requirements and contrasting reproductive modes. Climatic configuration determines distribution patterns and promotes sympatry of environmental niches, but specificity/selectivity determines the success of reproductive modes. Species distribution models (SDM) are a valuable tool to predict spatio-temporal distributions based on the extrapolation of environmental predictors. Objectives: To determine the spatio-temporal distribution of environmental niches and assess whether the protected areas of the World Database of Protected Areas (WDPA) allow the conservation of these species in the current scenario and future. Methods: We applied different algorithms to predict the distribution and spatio-temporal overlap of environmental niches of L. latinasus and P. cuqui within South America in the last glacial maximum (LGM), middle-Holocene, current and future scenarios. We assess the conservation status of both species with the WDPA conservation units. Results: All applied algorithms showed high performance for both species (TSS = 0.87, AUC = 0.95). The L. latinasus predictions showed wide environmental niches from LGM to the current scenario (49 % stable niches, 37 % gained niches, and 13 % lost niches), suggesting historical fidelity to stable climatic-environmental regions. In the current-future transition, L. latinasus would increase the number of stable (70 %) and lost (20 %) niches, suggesting fidelity to lowland regions and a possible trend toward microendemism. P. cuqui loses environmental niches from the LGM to the current scenario (25 %) and in the current-future transition (63 %), increasing the environmental sympathy between both species; 31 % spatial overlap in the current scenario and 70 % in the future. Conclusion: Extreme drought events and rainfall variations, derived from climate change, suggest the loss of environmental niches for these species that are not currently threatened but are not adequately protected by conservation units. The loss of environmental niches increases spatial sympatry which represents a new challenge for anurans and the conservation of their populations.

Resumen Introducción: Leptodactylus latinasus y Physalaemus cuqui son especies de anuros simpátricos con requerimientos ambientales similares y modos reproductivos contrastantes. La configuración climática determina los patrones de distribución y promueve la simpatría de los nichos ambientales, pero la especificidad/selectividad determina el éxito de los modos reproductivos. Los modelos de distribución de especies (MDE) son una herramienta valiosa para predecir distribuciones espacio-temporales basadas en la extrapolación de predictores ambientales. Objetivos: Determinar la distribución espacio-temporal de los nichos ambientales y evaluar si las áreas protegidas de la base de Datos Mundial de Áreas Protegidas (DMAP) permiten la conservación de estas especies en el escenario actual y futuro. Métodos: Aplicamos diferentes algoritmos para predecir la distribución y superposición espacio-temporal de nichos ambientales de L. latinasus y P. cuqui dentro de América del Sur en el último máximo glacial (UGM), Holoceno medio, actual y futuro. Evaluamos el estado de conservación de ambas especies con las unidades de conservación de la DMAP. Resultados: Todos los algoritmos aplicados mostraron un alto rendimiento para ambas especies (TSS = 0.87, AUC = 0.95). Las predicciones de L. latinasus mostraron amplios nichos ambientales desde LGM hasta el escenario actual (49 % de nichos estables, 37 % de nichos ganados y 13 % de nichos perdidos), sugiriendo fidelidad histórica por regiones climático-ambientales estables. En la transición actual-futura L. latinasus incrementaría la cantidad de nichos estables (70 %) y perdidos (20 %), sugiriendo fidelidad por regiones de tierras bajas y la posible tendencia hacia el microendemismo. P. cuqui pierde nichos ambientales desde el LGM al escenario actual (25 %) y en la transición actual-futura (63 %), incrementando la simpatría ambiental entre ambas especies; 31 % de superposición espacial en el escenario actual y 70 % en el futuro. Conclusión: Los eventos de sequía extrema y las variaciones de precipitaciones, derivados del cambio climático, sugieren la pérdida de nichos ambientales para estas especies, actualmente no se encuentran amenazadas, pero no están adecuadamente protegidas por las unidades de conservación. La pérdida de nichos ambientales aumenta la simpatría espacial que representa un nuevo desafío para estos anuros y la conservación de sus poblaciones.

Phylogenetic relationships of the Mangrove Hummingbird, "Amazilia" boucardi (Apodiformes: Trochilidae) of Costa Rica / Relaciones filogenéticas del Colibrí manglero "Amazilia" boucardi (Apodiformes: Trochilidae) de Costa Rica

Abstract Introduction: A recent revision of the generic classification of the Trochilidae based on DNA sequences revealed many inconsistencies with the current generic classification, largely based on plumage characters subject to homoplasy, especially in the Trochilini, the largest tribe. A thorough generic reorganization brought the classification into accord with the phylogeny, but due to lack of genetic data, two species remained unclassified. One of these was the Mangrove Hummingbird, "Amazilia" boucardi, endemic to Costa Rica and included in the IUCN red list of threatened species. Objective: To obtain molecular evidence to clarify the generic relationships of "A." boucardi. Methods: We isolated DNA from tissues of this species and amplified 4 nuclear and 4 mitochondrial fragments and compared these with homologous fragments from 56 species in the Trochilini, constructing phylogenetic trees with maximum likelihood and Bayesian methods. Results: Our phylogenetic analyses confirmed the placement of boucardi in the Trochilini and definitely excluded it from Amazilia but placed it with high confidence in the genus Chrysuronia Bonaparte, 1850, within which its closest relative is C. coeruleogularis, which also inhabits mangroves. Conclusions: Our genetic data based on nuclear and mitochondrial regions clearly indicate the relationship of A. boucardi and L. coeruleogularis. Moreover, it is also supported by their habitat distribution in the mangroves of the Pacific coast of Costa Rica and Western Panama. Therefore, we suggested to exclude A. boucardi as "incertae sedis".

Resumen Introducción: Una revisión reciente de la clasificación de la familia Trochilidae con base en secuencias de ADN demostró muchas incongruencias con la clasificación genérica previa, que había sido hecho con base en caracteres del plumaje muy sujetos a homoplasia, especialmente en la tribu más grande, Trochillini. Una reorganización de los géneros logró llevar su clasificación genérica a la concordancia con la filogenia, pero debido a la ausencia de datos genéticos, dos especies permanecieron sin clasificar. Una de estas fue el colibrí de manglar Amazilia boucardi, una especie endémica de Costa Rica, considerada como amenazada en la lista roja de la UICN. Objetivo: Obtener evidencia molecular para esclarecer las relaciones genéricas de A. boucardi. Métodos: Se aisló ADN de tejidos de esta especie y se amplificaron 4 fragmentos de ADN del núcleo y 5 de la mitocondria, y se compararon con fragmentos homólogos de 56 especies en la tribu Trochillini, generando árboles filogenéticos con métodos de máxima verosimilitud y bayesiano. Resultados: Los análisis filogénticos obtenidos confirmaron la ubicación de boucardi en Trochilini y definitivamente la excluyó del género Amazilia, pero la ubicó con un alto grado de confianza en el género Chrysuronia Bonaparte, 1850, dentro los cuales su pariente más cercano es C. coeruleogularis, que también habita manglares. Conclusiones: Nuestros datos genéticos basados en regiones nucleares y mitocondriales indican claramente la relación entre A. boucardi and L. coeruleogularis. Es más, lo anterior se sustenta por su distribución en los manglares de la costa Pacífica de Costa Rica y oeste de Panamá. Por lo tanto, sugerimos excluir a A. boucardi como "incertae sedis".

Impact of Collared Peccaries Dycotiles tajacu (Artiodactyla: Tayassuidae) on understory vegetation in the tropical rainforest of the Nogal-La Selva Biological Corridor, Costa Rica / Impacto del pecarí de collar, Dycotiles tajacu (Artiodactyla: Tayassuidae) en la vegetación del sotobosque del bosque tropical húmedo del Corredor Biológico Local Nogal-La Selva, Costa Rica

Abstract Introduction: Evidence suggests that herbivores, such as peccaries, shape vegetation structure and diversity through predation, trampling, dispersal, and rooting behavior. Objective: To evaluate the impact of peccaries (Dycotiles tajacu) on the understory vegetation of the tropical rainforest in the Nogal-La Selva Local Biological Corridor, Costa Rica, comparing a site with the absence of peccaries to another with the presence of these animals. Methodology: From June to November 2021, 20 experimental exclusions and 20 free access plots, each measuring 2 m2 were used to quantify herbivory, the number of leaf blades, damaged leaves, healthy leaves, sapling height, and fallen biomass at both sites. Results: A higher sapling density was found in the Nogal Reserve, but a lower sapling diversity, while in La Selva there was a higher sapling diversity, but a lower density of seedlings. Herbivory and sapling height in La Selva exceeded those in Nogal. The exclusion of peccaries reduced seedling damage but did not affect the dynamics of fallen biomass. Conclusion: For the design, implementation, and evaluation of the effectiveness of biological corridors, it is crucial to consider plant-animal interactions to enhance the flow of ecological processes through functional and structural connectivity, analyzed from interactions such as those presented in this paper.

Resumen Introducción: Existe evidencia que herbívoros, como los saínos, dan forma a la estructura y diversidad de la vegetación a través del comportamiento de depredación, pisoteo, dispersión y enraizamiento. Objetivo: Evaluar el impacto de los saínos (Dycotiles tajacu) en la vegetación del sotobosque del bosque tropical húmedo en el Corredor Biológico Local Nogal-La Selva, Costa Rica, en un sitio con ausencia y en otro con presencia de saínos. Métodos: De junio a noviembre de 2021 se utilizaron 20 exclusiones experimentales y 20 parcelas de acceso libre de 2 m2, se cuantifico la herbivoría, número de láminas foliares, hojas dañadas, hojas sanas, altura de brinzales y biomasa caída en ambos sitios. Resultados: Se encontró una mayor densidad de brinzales en Reserva Nogal pero una menor diversidad, contrario en La Selva donde se encontró una mayor diversidad de brinzales, pero una menor densidad de plántulas. La herbivoría y la altura de brinzales en La Selva fue mayor que en Nogal. La exclusión de los saínos disminuyó el daño a las plántulas, pero no afectó la dinámica de la biomasa caída. Conclusión: Es necesario contemplar para el diseño, implementación y evaluación de la efectividad de corredores biológicos, las interacciones planta-animal, para potencializar el flujo de procesos ecológicos mediante la conectividad funcional y estructural, analizada a partir de interacciones como las presentadas en este trabajo.

Diversidad de ninfas de Leptohyphidae (Ephemeroptera) en el río Quenane-Quenanito, piedemonte llanero colombiano / Diversity of Leptohyphidae (Ephemeroptera) nymphs in the Quenane-Quenanito river, Colombian plain foothills

Resumen Introducción: Varias presiones antrópicas sufren los ecosistemas acuáticos del piedemonte llanero en Colombia. La respuesta a estresores ambientales aún se desconoce en organismos bioindicadores como Leptohyphidae. Objetivo: Determinar la diversidad de ninfas de Leptohyphidae del río Quenane-Quenanito, en dos periodos hidrológicos contrastantes y su relación con algunas variables fisicoquímicas. Métodos: En diciembre (2014) y febrero (2015) se recolectaron organismos con red Surber en seis estaciones a lo largo del río. Se analizó la diversidad alfa y beta y se aplicó análisis de redundancia y modelos lineales generalizados con el fin de establecer la relación entre los taxones y las variables ambientales. Resultados: Se identificaron 369 organismos pertenecientes a cuatro géneros (Amanahyphes, Traverhyphes, Tricorythopsis y Tricorythodes), dos especies y ocho morfoespecies. Se reporta por primera vez para el departamento del Meta Amanahyphes saguassu. Se registró la mayor diversidad de ninfas en la transición a la sequía y la mayor abundancia en sequía. La diversidad beta señaló que la configuración del ensamblaje cambia a nivel espacial y temporal. Conclusiones: Los organismos de Leptohyphidae prefieren hábitats de corrientes, particularmente en el periodo de sequía, donde hallan alimento (hojarasca, detritos) y refugio para establecerse exitosamente; actividades antrópicas como la urbanización afectan notablemente la diversidad. La alta diversidad registrada en este pequeño río de piedemonte llanero refleja la necesidad de incrementar este tipo de trabajos y esfuerzos de recolección de material de estudio en la región.

Abstract Introduction: Various anthropic pressures affect the aquatic ecosystems of the foothills of Colombia. The response to environmental stressors is still unknown in bioindicator organisms such as Leptohyphidae. Objective: To determine the diversity of Leptohyphidae nymphs of the Quenane-Quenanito river, in two contrasting hydrological periods and its relationship with some physicochemical variables. Methods: In December (2014) and February (2015), organisms were collected with a Surber net at six stations along the current. Alpha and beta diversity was analyzed and redundancy analysis and generalized linear model were applied to establish the relationship between taxa and environmental variables. Results: Were identified 369 organisms belonging to four genera (Amanahyphes, Traverhyphes, Tricorythopsis, and Tricorythodes), two species, and eight morphospecies. Amanahyphes saguassu is reported for the first time for the Meta department. High diversity of Leptohyphidae nymphs was recorded in the transition to drought season and greater abundance in drought. Beta diversity indicated that the configuration of the assemblage changes spatially and temporally. Conclusions: Leptohyphidae organisms prefer fast habitats, particularly in the dry period where they find food (leaf litter, detritus) and shelter to establish themselves successfully; anthropic activities such as urbanization notably affect diversity. The high diversity recorded in this small river in the foothills of the plains reflects the need to increase this type of works and collection efforts of study material in the region.

Precipitation explains Wood Thrush (Hylocichla mustelina) occupancy patterns in Northern Costa Rica / Patrones de ocurrencia del Zorzal del Bosque (Hylocichla mustelina) explicados por la precipitación en el Norte de Costa Rica

Abstract Introduction: The Wood Thrush is a migratory bird that has experienced dramatic declines in its populations in recent decades. This species overwinters in forest fragments with intermediate levels of habitat modification in Central America. However, more studies detailing the use of remnant forests through time are needed to elucidate the threats this species faces in the wintering grounds. Objective: To understand the effects of environmental and forest structure variables on the occupancy of Wood Thrush in Northern Costa Rica. Methods: The study area was the Área de Conservación Guanacaste (ACG), located in Northern Costa Rica, in December 2016, and during the 2018-2019 migration season. We estimated Wood Thrush occupancy and detection probability in four locations of ACG (dry forest, cloud forest, and two locations in the wet forest) using single-season occupancy models. We also estimated Wood Thrush occupancy and probability of persistence in different months in three vegetation types (open area, secondary forest, and old-growth forest) in the wet forest of ACG using a multi-season occupancy model approach. Results: Wood Thrush occupancy was best described by precipitation in the four locations of the ACG; the probability of occupancy increased with precipitation. The average occupancy of Wood Thrushes varied with vegetation type: open area with shrubs and forest edge (0.69 ± 0.09), secondary forest (0.46 ± 0.1), and old-growth forest (0.61 ± 0.1). Wood Thrush probability of persistence responded partially to changes in precipitation, with an unexpected increase in persistence when the rainfall continued decreasing in the season. Conclusion: Wood Thrush occupancy was best predicted by changes in precipitation considering a larger spatial scale. Its probability of persistence partially varied with precipitation. An increase in persistence closer to Spring migration might be explained by the start of the breeding season of resident birds, potentially reducing territorial conflicts and conserving energy before migration. The long-term protection of wet forests in Northern Costa Rica is of paramount importance for the conservation of Wood Thrushes in their wintering grounds.

Resumen Introducción: El Zorzal del Bosque es un ave migratoria que ha experimentado caídas dramáticas en sus poblaciones en las últimas décadas. Esta especie pasa el invierno en fragmentos de bosque con niveles intermedios de modificación de hábitat en Centroamérica. Sin embargo, se necesitan más estudios que detallen el uso de los bosques remanentes a lo largo del tiempo para dilucidar las amenazas que enfrenta esta especie en las zonas de invernada. Objetivo: Comprender los efectos de variables ambientales y de estructura del bosque en la ocurrencia del Zorzal del Bosque en el Norte de Costa Rica. Métodos: El área de estudio fue el Área de Conservación Guanacaste (ACG), ubicada en el Norte de Costa Rica, en diciembre de 2016, y en la temporada migratoria 2018-2019. Estimamos la ocurrencia y la probabilidad de detección del Zorzal del Bosque en cuatro ubicaciones de ACG (bosque seco, bosque nuboso y dos ubicaciones en el bosque húmedo) utilizando modelos de ocurrencia de una sola temporada. También estimamos la ocurrencia del Zorzal del Bosque y la probabilidad de persistencia en diferentes meses en tres tipos de vegetación (área abierta, bosque secundario y bosque primario) en el bosque húmedo de ACG utilizando un enfoque de modelo de ocurrencia multi-estacional. Resultados: La ocurrencia del Zorzal del Bosque estuvo mejor descrita por la precipitación en las cuatro localidades del ACG; la probabilidad de ocurrencia aumentó con las precipitaciones. La ocurrencia media de zorzales varió con el tipo de vegetación: área abierta con arbustos y borde de bosque (0.69 ± 0.09), bosque secundario (0.46 ± 0.1) y bosque primario (0.61 ± 0.1). La probabilidad de persistencia del zorzal respondió parcialmente a cambios en la precipitación, con un aumento inesperado en la persistencia cuando las precipitaciones continuaron disminuyendo en la temporada. Conclusión: La ocurrecia del Zorzal del Bosque varió con la precipitación considerando una escala espacial mayor. Su probabilidad de persistencia varió parcialmente con la precipitación. Un aumento en la persistencia más cerca de la migración de primavera podría explicarse por el inicio de la temporada de reproducción de las aves residentes, lo que podría reducir los conflictos territoriales y conservar energía antes de la migración. La protección a largo plazo de los bosques húmedos en el norte de Costa Rica es de suma importancia para la conservación de los Zorzales del Bosque en sus zonas de invernada.

Assessment of heavy metal levels in polyculture fish farms and their aquatic ecosystems: an integrative study addressing environmental and human health risks associated with dam water usage.

This study examines the levels of heavy metals in polyculture fish (Labeo rohita, Cyprinus carpio, and Catla catla), water, and sediment in Tanda Dam, Kohat, Pakistan, aiming to understand environmental and health risks. Samples of fish, water, and sediment were collected from 3 fish farms, and heavy metal concentrations were measured using a Flame Atomic Absorption Spectrophotometer (AAS). Results reveal that C. catla exhibited significantly higher (p < 0.05) levels of Zn than other fish species. Conversely, C. carpio showed significantly higher (p < 0.05) concentrations of Pb, Cd, Cr, Mn, Cu, As, and Ni than other species. The heavy metal hierarchy in C. carpio was found to be Zn > Cu > Pb > Cr > Cd > Mn > As > Ni. While heavy metal levels in L. rohita and C. catla generally fell within reference ranges, exceptions were noted for Zn, Pb, and Cd. Conversely, in C. carpio, all metals exceeded reference ranges except for Cu and Ni. Principal Component Analysis (PCA) indicated a close relationship between water and sediment. Additionally, cluster analysis suggested that C. catla formed a distinct cluster from L. rohita and C. carpio, implying different responses to the environment. Despite concerns raised by the Geoaccumulation Index (Igeo) and Contamination Factor (CF), particularly for Cd, which exhibited a high CF. Furthermore, Hazard Index (HI) values for all three fish species were below 1, suggesting low health risks. However, elevated Igeo and CF values for Cd suggest significant pollution originating from anthropogenic sources. This study underscores the importance of monitoring heavy metals in water for both environmental preservation and human health protection. Future research efforts should prioritize pollution control measures to ensure ecosystem and public health safety.

The GE296_RS03820 and GE296_RS03830 genes are involved in capsular polysaccharide biosynthesis in Riemerella anatipestifer.

Riemerella anatipestifer is a pathogenic bacterium that causes duck serositis and meningitis, leading to significant harm to the duck industry. To escape from the host immune system, the meningitis-causing bacteria must survive and multiply in the bloodstream, relying on specific virulence factors such as capsules. Therefore, it is essential to study the genes involved in capsule biosynthesis in R. anatipestifer. In this study, we successfully constructed gene deletion mutants Δ3820 and Δ3830, targeting the GE296_RS03820 and GE296_RS03830 genes, respectively, using the RA-LZ01 strain as the parental strain. The growth kinetics analysis revealed that these two genes contribute to bacterial growth. Transmission and scanning electron microscopy (TEM and SEM) and silver staining showed that Δ3820 and Δ3830 produced the altered capsules and compounds of capsular polysaccharides (CPSs). Serum resistance test showed the mutants also exhibited reduced C3b deposition and decreased resistance serum killing. In vivo, Δ3820 and Δ3830 exhibited markedly declining capacity to cross the blood-brain barrier, compared to RA-LZ01. These findings indicate that the GE296_RS03820 and GE296_RS03830 genes are involved in CPSs biosynthesis and play a key role in the pathogenicity of R. anatipestifer. Furthermore, Δ3820 and Δ3830 mutants presented a tendency toward higher survival rates from RA-LZ01 challenge in vivo. Additionally, sera from ducklings immunized with the mutants showed cross-immunoreactivity with different serotypes of R. anatipestifer, including 1, 2, 7 and 10. Western blot and SDS-PAGE assays revealed that the altered CPSs of Δ3820 and Δ3830 resulted in the exposure of some conserved proteins playing the key role in the cross-immunoreactivity. Our study clearly demonstrated that the GE296_RS03820 and GE296_RS03830 genes are involved in CPS biosynthesis in R. anatipestifer and the capsule is a target for attenuation in vaccine development.

Transitions in development - an interview with Sonia Sen.

Sonia Sen is Group Leader at the Tata Institute for Genetics and Society in Bangalore, India. Her group is interested in understanding neurodevelopment and behaviour in Drosophila and Anopheles. We spoke to Sonia over a video call to talk about her research career, her transition to becoming a group leader, and the work that the Indian Society of Developmental Biologists is doing.

Engineered mini-G proteins block the internalization of cognate GPCRs and disrupt downstream intracellular signaling.

Mini-G proteins are engineered, thermostable variants of Gα subunits designed to stabilize G protein-coupled receptors (GPCRs) in their active conformations. Because of their small size and ease of use, they are popular tools for assessing GPCR behaviors in cells, both as reporters of receptor coupling to Gα subtypes and for cellular assays to quantify compartmentalized signaling at various subcellular locations. Here, we report that overexpression of mini-G proteins with their cognate GPCRs disrupted GPCR endocytic trafficking and associated intracellular signaling. In cells expressing the Gαs-coupled GPCR glucagon-like peptide 1 receptor (GLP-1R), coexpression of mini-Gs, a mini-G protein derived from Gαs, blocked ß-arrestin 2 recruitment and receptor internalization and disrupted endosomal GLP-1R signaling. These effects did not involve changes in receptor phosphorylation or lipid nanodomain segregation. Moreover, we found that mini-G proteins derived from Gαi and Gαq also inhibited the internalization of GPCRs that couple to them. Finally, we developed an alternative intracellular signaling assay for GLP-1R using a nanobody specific for active Gαs:GPCR complexes (Nb37) that did not affect GLP-1R internalization. Our results have important implications for designing methods to assess intracellular GPCR signaling.

Insulated by opioids.

Opioids trigger myelin insulation of reward circuit axons in a feedforward loop of addiction.

Revisiting Perdeck's massive avian migration experiments debunks alternative social interpretations.

Whether avian migrants can adapt to their changing world depends on the relative importance of genetic and environmental variation for the timing and direction of migration. In the classic series of field experiments on avian migration, A. C. Perdeck discovered that translocated juveniles failed to reach goal areas, whereas translocated adults performed 'true-goal navigation'. His translocations of > 14 000 common starlings (Sturnus vulgaris) suggested that genetic mechanisms guide juveniles into a population-specific direction, i.e. 'vector navigation'. However, alternative explanations involving social learning after release in juveniles could not be excluded. By adding historical data from translocation sites, data that was unavailable in Perdeck's days, and by integrated analyses including the original data, we could not explain juvenile migrations from possible social information upon release. Despite their highly social behaviour, our findings are consistent with the idea that juvenile starlings follow inherited information and independently reach their winter quarters. Similar to more solitarily migrating songbirds, starlings would require genetic change to adjust the migration route in response to global change.

Earliest evidence of avian primary feather moult.

Feather moulting is a crucial process in the avian life cycle, which evolved to maintain plumage functionality. However, moulting involves both energetic and functional costs. During moulting, plumage function temporarily decreases between the shedding of old feathers and the full growth of new ones. In flying taxa, a gradual and sequential replacement of flight feathers evolved to maintain aerodynamic capabilities during the moulting period. Little is known about the moult strategies of non-avian pennaraptoran dinosaurs and stem birds, before the emergence of crown lineage. Here, we report on two Early Cretaceous pygostylian birds from the Yixian Formation (125 mya), probably referable to Confuciusornithiformes, exhibiting morphological characteristics that suggest a gradual and sequential moult of wing flight feathers. Short primary feathers interpreted as immature are symmetrically present on both wings, as is typical among extant flying birds. Our survey of the enormous collection of the Tianyu Museum confirms previous findings that evidence of active moult in non-neornithine pennaraptorans is rare and likely indicates a moult cycle greater than one year. Documenting moult in Mesozoic feathered dinosaurs is critical for understanding their ecology, locomotor ability and the evolution of this important life-history process in birds.