Recent molecular techniques to strengthen ecological studies in echinoderms / Técnicas moleculares recientes que fortalecen los estudios ecológicos de equinodermos

Abstract Introduction: Echinoderms, an integral component of marine ecosystems worldwide, have captivated scientific interest for centuries. Despite this longstanding attention, comprehending key facets such as trophic relationships, diet composition, and host-microbiota relationships still represents a challenge using traditional techniques. Recent years, however, have witnessed a transformative shift, thanks to the emergence of advanced molecular techniques, offering new approaches to strengthen ecological studies in echinoderms. Objective: Explore how recent advancements in molecular tools have impacted ecological research on echinoderms. Specifically, we aim to investigate the potential of these tools to shed light on trophic interactions, diet composition, and the characterization of gut microbial communities in these organisms. Methods: Available literature was used to clarify how novel molecular techniques can improve ecological studies. The focus is diet, trophic relationships, and gut microbiota. Results: Traditionally, studies of stomach contents using compound microscopy have provided an idea of ingested material; nevertheless, sometimes a simple magnified visualization of dietary content does not allow exhaustive identification of the entire food spectrum, as it is limited due to the rapid digestion and maceration of food items within the echinoderm's digestive tract. The use of DNA-metabarcoding, targeting specific DNA regions, such as the mitochondrial COI gene, has allowed us to enhance the accuracy and precision of diet characterization by enabling the identification of prey items down to the species or even genetic variant level, providing valuable insights into specific dietary preferences. Another approach is the use of stable isotopes, particularly carbon and nitrogen, which provide a powerful tool to trace the origin and flow of nutrients through food webs. By analyzing the isotopic signatures in muscular tissues and food items, we can discern the sources of their primary food items and gain insights into their trophic position within the ecosystem. Lastly, a third new technique used to elucidate the characterization of the prokaryotic community is 16S rRNA sequencing. This method allows us to explore the composition and dynamics of the digestive tract microbial communities. Conclusions: This is a promising era for ecological research on echinoderms, where advances of molecular tools have enabled an unprecedented level of detail, resolving longstanding challenges in comprehending their trophic interactions, diet composition, and host-microbiota relationships, and opening new avenues of investigation in ecological studies.

Resumen Introducción: Los equinodermos, un componente integral de los ecosistemas marinos en todo el mundo, han captado el interés científico durante siglos. A pesar de esta prolongada atención, el comprender facetas clave como las relaciones tróficas, la composición de la dieta y las relaciones huésped-microbiota todavía representa un desafío utilizando técnicas tradicionales. Sin embargo, los últimos años han sido testigos de un cambio transformador, gracias a la aparición de técnicas moleculares avanzadas, que ofrecen nuevos enfoques para fortalecer los estudios ecológicos en equinodermos. Objetivo: Explorar cómo los avances recientes en herramientas moleculares han impactado la investigación ecológica sobre equinodermos. Específicamente, nuestro objetivo es investigar el potencial de estas herramientas para arrojar luz sobre las interacciones tróficas, la composición de la dieta y la caracterización de las comunidades microbianas intestinales en estos organismos. Métodos: Se utilizó la literatura disponible para aclarar cómo las nuevas técnicas moleculares pueden mejorar los estudios ecológicos. La atención se centra en la dieta, las relaciones tróficas y la microbiota intestinal. Resultados: Tradicionalmente, los estudios del contenido estomacal mediante microscopía compuesta han proporcionado una idea del material ingerido; Sin embargo, a veces una simple visualización ampliada del contenido dietético no permite una identificación exhaustiva de todo el espectro alimentario, ya que está limitado debido a la rápida digestión y maceración de los alimentos dentro del tracto digestivo del equinodermo. El uso de metabarcoding de ADN, dirigidos a regiones específicas del ADN, como el gen COI mitocondrial, nos ha permitido mejorar la exactitud y precisión de la caracterización de la dieta al permitir la identificación de presas hasta el nivel de especie o incluso de variante genética, lo que proporciona valiosos resultados sobre preferencias dietéticas específicas. Otro enfoque es el uso de isótopos estables, en particular carbono y nitrógeno, que proporcionan una poderosa herramienta para rastrear el origen y el flujo de nutrientes a través de las redes alimentarias. Al analizar las firmas isotópicas en los tejidos musculares y los alimentos, podemos discernir las fuentes de sus alimentos primarios y obtener información sobre su posición trófica dentro del ecosistema. Por último, una tercera técnica nueva utilizada para dilucidar la caracterización de la comunidad procariótica es la secuenciación del ARNr 16S. Este método nos permite explorar la composición y dinámica de las comunidades microbianas del tracto digestivo. Conclusiones: Esta es una era prometedora para la investigación ecológica sobre equinodermos, donde los avances de las herramientas moleculares han permitido un nivel de detalle sin precedentes, resolviendo desafíos de larga data en la comprensión de sus interacciones tróficas, composición de la dieta y relaciones huésped-microbiota, y abriendo nuevas vías de investigación. en estudios ecológicos.

Uneven demographic consequences of the 2022 disease outbreak for the sea urchin Diadema antillarum in Puerto Rico.

Pervasive epizootic events have had a significant impact on marine invertebrates throughout the Caribbean, leading to severe population declines and consequential ecological implications. One such event was the regional collapse of herbivory, partly caused by the Diadema antillarum mortality event in 1983-84, resulting in a trophic cascade and altering the structure of reef communities. Consequently, there was a notable decrease in coral recruitment and an increase in the coverage of macroalgae. Nearly four decades later, in early 2022, the Caribbean basin experienced another widespread mass mortality event, further reducing the populations of D. antillarum. To assess the effects of this recent mortality event on the current demographics of D. antillarum, we surveyed eight populations along the eastern, northeastern, northern, and northwestern coast of Puerto Rico from May to July 2022, estimating their population density, size distribution, and disease prevalence. Additionally, the study compared these population parameters with data from four sites previously surveyed in 2012 and 2017 to understand the impact of the recent mortality event. The survey conducted in 2022 showed varying population densities at the surveyed reefs. Some populations exhibited mean densities of nearly one individual per square meter, while others had extremely low or no living individuals per square meter. The four populations with the highest density showed no evidence of disease, whereas the four populations with the lowest D. antillarum densities exhibited moderate to high disease prevalence. However, when considering all sites, the estimated disease prevalence remained below 5%. Nevertheless, the comparison with data from 2012 and 2017 indicated that the recent mortality event had a negative impact on D. antillarum demographics at multiple sites, as the densities in 2022 were reduced by 60.19% compared to those from the previous years. However, it is still too early to determine the severity of this new mortality event compared to the 1983-84 mortality event. Therefore, it is imperative to continue monitoring these populations.

Aprendizaje creativo en estudiantes de Medicina desde la Genética Médica

Fundamento: En la carrera de Medicina es necesario formar profesionales con pensamiento creativo que permita la solución de problemas vinculados con el ejercicio de su profesión. La Genética Médica aporta la posibilidad de reflexionar y crear soluciones creativas. Objetivo: Proponer actividades que posibiliten el aprendizaje creativo en estudiantes de Medicina desde la Genética Médica. Metodología: Se emplearon métodos teóricos, empíricos y matemático-estadístico, con una muestra de 34 estudiantes y 3 profesores que imparten la asignatura. Resultados: Los docentes reconocen que, a pesar de hacer intentos para alcanzar el aprendizaje creativo, desconocen algunas vías y manifiestan no contar con el tiempo suficiente para su logro. El accionar didáctico no favorece el conocimiento, aplicación y disposición de estudiantes para un aprendizaje creativo. Se propusieron actividades sustentadas en procederes en Genética Médica, como parte de la estrategia didáctica de los docentes, asimismo contribuir al perfeccionamiento del aprendizaje creativo en los estudiantes. Las mismas centradas en métodos problémicos y el uso de recursos tecnológicos. Conclusiones: La formación de los médicos necesita expresar un aprendizaje creativo vinculado a un desempeño eficiente en la solución de problemas de la comunidad, lo cual se puede facilitar mediante la utilización de actividades en la asignatura Genética Médica.

Background: In the Medicine career it is necessary to form professionals with creative thoughts that allow the solution of problems linked to their professional practice. Medical Genetics provides the possibility to reflect and create creative solutions. Objective: To propose activities that allow creative learning in Medical students from the Medical Genetics perspective. Methodology: Theoretical, empirical and mathematical-statistical methods were applied, with a sample of 34 students and 3 teachers who teach the subject. Results: Teachers recognize that, despite making attempts to achieve creative learning, they are unaware of some ways and declare that they do not have enough time to achieve it. The didactic action does not favor the knowledge, application and disposition of students for creative learning. Activities supported by procedures in Medical Genetics were proposed as part of the didactic strategy of the teachers, likewise contribute to the improvement of creative learning in students. The same ones are focused on problem-solving methods and the use of technological resources. Conclusions: Physician education needs to express creative learning coupled with efficient performance in solving community problems, which may be facilitated through the use of activities in Medical Genetics Medical subject.

The Epibiotic Microbiota of Wild Caribbean Sea Urchin Spines Is Species Specific.

Caribbean sea urchins are marine invertebrates that have experienced a decline over the years. Studies on sea urchins have focused primarily on the microbiome of the coelomic fluid or the gut microbiota. In this study, the epibiota community associated with four wild Caribbean sea urchin species, Lytechinus variegatus, Echinometra lucunter, Tripneustes ventricosus, and Diadema antillarum, was characterized for the first time. Using 57 sea urchin animal samples, we evaluated the influence of animal species, trophic niches, and geographical location on the composition of the epibiotic microbiota. We found significant differences in the bacterial biota among species and trophic niches, but not among geographical locations. L. variegatus exhibited the highest alpha diversity with high dominance of Fusobacteria, Planctomycetes, and Cyanobacteria, whereas T. ventricosus and D. antillarum were dominated by Firmicutes. T. ventricosus inhabiting the seagrass biotope dominated by Thalassia testudinum meadows had mostly Endozoicomonas. In contrast, samples located in the reef (dominated by corals and other reef builders) had a higher abundance of Kistimonas and Photobacterium. Our findings confirm that the epibiotic microbiota is species-specific, but also niche-dependent, revealing the trophic networks emerging from the organic matter being recycled in the seagrass and reef niches. As echinoids are important grazers of benthic communities, their microbiota will likely influence ecosystem processes.

Trophic niches reflect compositional differences in microbiota among Caribbean sea urchins.

Sea urchins play a critical role in marine ecosystems, as they actively participate in maintaining the balance between coral and algae. We performed the first in-depth survey of the microbiota associated with four free-living populations of Caribbean sea urchins: Lytechinus variegatus, Echinometra lucunter, Tripneustes ventricosus, and Diadema antillarum. We compared the influence of the collection site, echinoid species and trophic niche to the composition of the microbiota. This dataset provides a comprehensive overview to date, of the bacterial communities and their ecological relevance associated with sea urchins in their natural environments. A total of sixty-samples, including surrounding reef water and seagrass leaves underwent 16S rRNA gene sequencing (V4 region) and high-quality reads were analyzed with standard bioinformatic approaches. While water and seagrass were dominated by Cyanobacteria such as Prochlorococcus and Rivularia respectively, echinoid gut samples had dominant Bacteroidetes, Proteobacteria and Fusobacteria. Propionigenium was dominant across all species' guts, revealing a host-associated composition likely responsive to the digestive process of the animals. Beta-diversity analyses showed significant differences in community composition among the three collection sites, animal species, and trophic niches. Alpha diversity was significantly higher among L. variegatus samples compared to the other species. L. variegatus also displayed an increased abundance of Planctomycetes and Cyanobacterial OTUs. The bacterial community of this herbivorous echinoid reflected similarities to the microfilm community found on Thalassia testudinum leaves; a very abundant seagrass and its main food resource. The results of this study elaborate on the microbial ecology of four important Caribbean echinoids, confirming that selection on the microbial community is trophic-niche dependent.

Understanding trophic relationships among Caribbean sea urchins

ResumenLas especies Echinometra lucunter, Echinometra viridis, Lytechinus variegatus, Tripneustes ventricosus, and Diadema antillarum son los erizos de mar más comunes en los hábitat litorales del Caribe. Los erizos de mar T. ventricosus y L. variegatus habitan generalmente los pastos marinos mientras que las otras tres especies se encuentran asociadas a sustratos rocosos. Los hábitos alimentarios de estas especies han sido bien documentados y son reconocidas como herbívoros - omnívoros; sin embargo, pocas de estas especies han sido caracterizadas isotópicamente. Utilizamos los isótopos estables para caracterizar estas cinco especies de erizos y establecer las posiciones tróficas para las especies que cohabitan los mismos ecosistemas. También cuantificamos la contribución de los recursos alimentarios para E. lucunter. Los erizos T. ventricosus y D. antillarum mostraron los mayores valores de δ15N y valores similares de δ13C que variaron desde -11.6 ± 0.63 a -10.4 ± 0.99 %; donde el erizo E. lucunter mostró los valores más negativos con -15.40 ± 0.76 %. Las comunidades de algas no mostraron diferencias en valores promedio de δ15N (F= 1.300, df= 3, p= 0.301), pero sí mostraron variaciones en los valores de δ13C (F= 7.410, df= 3, p= 0.001). Los estudios de amplitud de elipses de nicho determinaron que las especies de los biotopos rocosos (D. antillarum, E. lucunter y E. viridis) no mostraron solapamiento de nicho. Similar resultado también se encontró en las especies de erizos que habitan en los pastos marinos. Sin embargo, la distancia entre estas dos especies fue menor respecto a la distancia entre las especies de erizos que habitan en los sustratos rocosos. Nuestros resultados muestran que las especies que habitan en los pastos marinos mostraron valores más elevados de δ13C en comparación con las especies de los sustratos rocosos. No se encontraron diferencias espaciales para E. lucunter en δ15N, pero sí en δ13C. Los modelos de mezcla bayesianos demuestran la plasticidad alimentaria de E. lucunter, especie capaz de utilizar múltiples recursos algales dependiendo de la disponibilidad por sitio. Semejanzas en δ15N entre D. antillarum y T. ventricosus parecen indicar similitudes tróficas entre ambas especies. Si bien T. ventricosus es reconocido como omnívoro, D. antillarum siempre ha sido considerado un herbívoro generalista. Finalmente, la falta de solapamiento entre las especies en los dos biotopos parece indicar una estrategia de partición de recursos para evitar la competencia de nicho entre especies concurrentes.

AbstractThe species Echinometra lucunter, Echinometra viridis, Lytechinus variegatus, Tripneustes ventricosus, and Diadema antillarum are the most common sea urchins of littoral habitats in the Caribbean. T. ventricosus and L. variegatus are associated with seagrass beds, while the other three species usually inhabit hardground substrates. Food preferences of these species are well documented and they are commonly accepted as being primarily herbivorous-omnivorous; nevertheless, few of them have previously been characterized isotopically. We used this approach for assessing the isotopic characterization of five echinoids. We established the trophic position of two groups of co-occurring species and quantified the contribution of food resources in the diet of Echinometra lucunter, considered the most common sea urchin in the Caribbean region. The species T. ventricosus and D. antillarum showed the highest values of δ15N. Sea urchins exhibited similar values of δ13C varying from -11.6 ± 0.63 to -10.4 ± 0.99%. The echinoid E. lucunter displayed the lowest values of carbon, from -15.40 ± 0.76%. Significant differences among species were found for δ15N and δ13C. Seaweed communities exhibited no differences among sites for overall δ15N (F= 1.300, df= 3, p= 0.301), but we found spatial differences for δ13C (F= 7.410, df= 3, p= 0.001). The ellipse-based metrics of niche width analysis found that the hardground biotope species (D. antillarum, E. lucunter, and E. viridis) did not overlap each other. Similar results were obtained for the co-occurring species of the seagrass biotope; however, the distance between these species was closer than that of the hardground biotope species. The Bayesian mixing models run for E. lucunter at all four localities found differences in food resources contribution. The algae D. menstrualis, C. crassa and B. triquetrum dominated in CGD; whereas C. nitens, Gracilaria spp., and D. caribaea represented the main contributor algae to the diet of E. lucunter at LQY. In Culebra Island, no dominance of any particular algae was detected in TMD, where six of the eight species exhibited a similar contribution. Similarities in δ15N between D. antillarum and T. ventricosus may hint towards a similar trophic level for these species, although T. ventricosus is widely accepted as an omnivore, while D. antillarum is considered a generalist herbivore. The lack of overlap among species in the two biotopes seems to indicate a resource partitioning strategy to avoid niche competition among co-occurring species. Rev. Biol. Trop. 64 (2): 837-848. Epub 2016 June 01.

Trophic ecology of sea urchins in coral-rocky reef systems, Ecuador.

Sea urchins are important grazers and influence reef development in the Eastern Tropical Pacific (ETP). Diadema mexicanum and Eucidaris thouarsii are the most important sea urchins on the Ecuadorian coastal reefs. This study provided a trophic scenario for these two species of echinoids in the coral-rocky reef bottoms of the Ecuadorian coast, using stable isotopes. We evaluated the relative proportion of algal resources assimilated, and trophic niche of the two sea urchins in the most southern coral-rocky reefs of the ETP in two sites with different disturbance level. Bayesian models were used to estimate the contribution of algal sources, niche breadth, and trophic overlap between the two species. The sea urchins behaved as opportunistic feeders, although they showed differential resource assimilation. Eucidaris thouarsii is the dominant species in disturbed environments; likewise, their niche amplitude was broader than that of D. mexicanum when conditions were not optimal. However, there was no niche overlap between the species. The Stable Isotope Analysis in R (SIAR) indicated that both sea urchins shared limiting resources in the disturbed area, mainly Dictyota spp. (contributions of up to 85% for D. mexicanum and up to 75% for E. thouarsii). The Stable Isotope Bayesian Ellipses in R (SIBER) analysis results indicated less interspecific competition in the undisturbed site. Our results suggested a trophic niche partitioning between sympatric sea urchin species in coastal areas of the ETP, but the limitation of resources could lead to trophic overlap and stronger habitat degradation.

Understanding trophic relationships among Caribbean sea urchins.

The species Echinometra lucunter, Echinometra viridis, Lytechinus variegatus, Tripneustes ventricosus, and Diadema antillarum are the most common sea urchins of littoral habitats in the Caribbean. T. ventricosus and L. variegatus are associated with seagrass beds, while the other three species usually inhabit hardground substrates. Food preferences of these species are well documented and they are commonly accepted as being primarily herbivorous-omnivorous; nevertheless, few of them have previously been characterized isotopically. We used this approach for assessing the isotopic characterization of five echinoids. We established the trophic position of two groups of co-occurring species and quantified the contribution of food resources in the diet of Echinometra lucunter, considered the most common sea urchin in the Caribbean region. The species T. ventricosus and D. antillarum showed the highest values of δ15N. Sea urchins exhibited similar values of δ13C varying from -11.6 ± 0.63 to -10.4 ± 0.99%. The echinoid E. lucunter displayed the lowest values of carbon, from -15.40 ± 0.76%. Significant differences among species were found for δ15N and δ13C. Seaweed communities exhibited no differences among sites for overall δ15N (F= 1.300, df= 3, p= 0.301), but we found spatial differences for δ13C (F= 7.410, df= 3, p= 0.001). The ellipse-based metrics of niche width analysis found that the hardground biotope species (D. antillarum, E. lucunter, and E. viridis) did not overlap each other. Similar results were obtained for the co-occurring species of the seagrass biotope; however, the distance between these species was closer than that of the hardground biotope species. The Bayesian mixing models run for E. lucunter at all four localities found differences in food resources contribution. The algae D. menstrualis, C. crassa and B. triquetrum dominated in CGD; whereas C. nitens, Gracilaria spp., and D. caribaea represented the main contributor algae to the diet of E. lucunter at LQY. In Culebra Island, no dominance of any particular algae was detected in TMD, where six of the eight species exhibited a similar contribution. Similarities in δ15N between D. antillarum and T. ventricosus may hint towards a similar trophic level for these species, although T. ventricosus is widely accepted as an omnivore, while D. antillarum is considered a generalist herbivore. The lack of overlap among species in the two biotopes seems to indicate a resource partitioning strategy to avoid niche competition among co-occurring species.