Molecular cloning, functional characterization and differential expression of two novel GABAAR-like subunits from red swamp crayfish Procambarus clarkii.

In this work, we cloned and functionally expressed two novel GABAA receptor subunits from Procambarus clarkii crayfish. These two new subunits, PcGABAA-α and PcGABAA-ß2, revealed significant sequence homology with the PcGABAA-ß subunit, previously identified in our laboratory. In addition, PcGABAA-α subunit also shared a significant degree of identity with the Drosophila melanogaster genes DmGRD (GABA and glycine-like receptor subunits of Drosophila) as well as PcGABAA-ß2 subunit with DmLCCH3 (ligand-gated chloride channel homolog 3). Electrophysiological recordings showed that the expression in HEK cells of the novel subunits, either alone or in combination, failed to form functional homo- or heteromeric receptors. However, the co-expression of PcGABAA-α with PcGABAA-ß evoked sodium- or chloride-dependent currents that accurately reproduced the time course of the GABA-evoked currents in the X-organ neurons from crayfish, suggesting that these GABA subunits combine to form two types of GABA receptors, one with cationic selectivity filter and the other preferentially permeates anions. On the other hand, PcGABAA-ß2 and PcGABAA-ß co-expression generated a chloride current that does not show desensitization. Muscimol reproduced the time course of GABA-evoked currents in all functional receptors, and picrotoxin blocked these currents; bicuculline did not block any of the recorded currents. Reverse transcription polymerae chain reaction (RT-PCR) amplifications and FISH revealed that PcGABAA-α and PcGABAA-ß2 are predominantly expressed in the crayfish nervous system. Altogether, these findings provide the first evidence of a neural GABA-gated cationic channel in the crayfish, increasing our understanding of the role of these new GABAA receptor subunits in native heteromeric receptors.

Epibiotic ciliate communities on the crayfish Procambarus (Austrocambarus) sp. cultivated in a rustic aquaculture in southern Mexico.

Rustic aquaculture systems represent a sustainable production alternative in the world, providing appropriate conditions for the establishment of epibiotic ciliates on crayfish. However, the impact of epibiotic ciliates on the aquaculture production has been insufficiently studied hitherto. The objective of this work was to identify the epibiotic ciliates on the surface of the crayfish Procambarus (Austrocambarus) sp. and analyze whether there is any correlation between seasonality and environmental conditions in an artificial pond in Chiapas, Mexico. The crayfish were collected during dry and rainy seasons to investigate their ciliate epibionts. Some physical and chemical parameters of the sampled water were measured and correlation matrix analyses were applied to detect possible relationships between the presence of ciliates and environmental parameters. Thirteen species belonging to the subclasses Peritrichia and Suctoria were identified. Significant differences were found for some environmental parameters and species recorded in both seasons. The present data could help to better understand the relationship bewteen the epibiotic ciliate community on crayfish and some environmental parameters in aquaculture systems.

The impact of Roundup® Original DI on the hemolymph metabolism and gill and hepatopancreas oxidative balance of Parastacus promatensis (Crustacea, Decapoda, Parastacidae).

In Brazil, glyphosate is present in more than 130 commercial formulations, and its toxic effects have already been tested in different species to understand its impact on biota Decapod crustaceans are widely used as experimental models due to their biology, sensitivity to pollutants, ease of collection, and maintenance under laboratory conditions. We evaluated the changes in metabolism (hemolymph) and oxidative balance markers (gill and hepatopancreas) of a crayfish (Parastacus promatensis) after exposure to Roundup® (active ingredient: glyphosate). The crayfish were captured in the Garapiá stream within the Center for Research and Conservation of Nature Pró-Mata, Brazil. We collected adult animals outside (fall) and during (spring) the breeding season. The animals were transported in buckets with cooled and aerated water from the collection site to the aquatic animal maintenance room at the university. After acclimatization, the animals were exposed to different concentrations of glyphosate (0, 65, 260, 520, and 780 µg/L). The results showed a significant variation in the hemolymph glucose, lactate, and protein levels. We observed variations in the tissue antioxidant enzymatic activity after exposure to glyphosate. Finally, the increase in oxidative damage required a high energy demand from the animals to maintain their fitness, which makes them more vulnerable to stress factors added to the habitat.

Fatty acid composition and metabolic pathways in Arbacia dufresnii (Arbaciidae: Arbacioida) gametes: implications of shrimp byproducts in aquaculture feeds / Composición de ácidos grasos y vías metabólicas en los gametos de Arbacia dufresnii (Arbaciidae: Arbacioida) implicaciones de los subproductos de langostinos en los alimentos de acuicultura

Abstract Introduction: Care towards nutrition is essential for the quality of a sustainable aquaculture product. Since the balance in food affects the growth and production of gametes. The circular economy is made possible through the use of discarded materials. Objective: The aim of this research was to study the fatty acid composition and metabolic pathways in the gametes of Arbacia dufresnii, with a focus on the implications of incorporating shrimp byproducts into aquaculture feeds. Methods: Four different treatments were designed to maintain optimal nutritional quality, particularly in lipids and proteins, based on previous studies. The fatty acid profiles of the feeds and gametes were analyzed by using gas-chromatography, and statistical analyses were conducted to determine significant differences. Results: Significant differences were observed in the abundance (%) of omega-3 (ω-3) and omega-6 (ω-6) fatty acids. The (ω-3) metabolic pathway was more pronounced in the gametes of wild animals and those fed with the experimental feeds. In contrast, the (ω-6) metabolic pathway was less relevant in these groups. The (ω-3) /(ω-6) ratio was highest in the gametes of wild animals. Feeds enriched in fatty acids enhanced their bioaccumulation in the gametes reaching higher concentrations than wild animals. The availability of fatty acids in foods allowed their bioaccumulation in gametes, with concentrations equal to or higher than those observed in animals in their natural environment for certain fatty acids. Conclusions: Incorporating shrimp byproducts in aquaculture feeds demonstrated a promising strategy for resource utilization and organic input generation. The fatty acid composition in the gametes of A. dufresnii was influenced by the diet, highlighting the potential of balanced feeds to enhance the bioaccumulation of essential fatty acids. These findings provide valuable insights for the development of sustainable aquaculture practices and the production of nutritionally enriched seafood products.

Resumen Introducción: El cuidado hacia la nutrición es fundamental para la calidad de un producto acuícola sostenible. Ya que el balance en los alimentos afecta el crecimiento y producción de los gametos. A partir del aprovechamiento de materias de descarte se posibilita la economía circular. Objetivo: El objetivo de este estudio fue investigar la composición de ácidos grasos y las vías metabólicas en los gametos de Arbacia dufresnii, centrándose en las implicaciones de la incorporación de subproductos de camarones en los alimentos de acuicultura. Métodos: Se diseñaron cuatro tratamientos diferentes para mantener una calidad nutricional óptima, especialmente en lípidos y proteínas, basándose en estudios previos. Se analizaron los perfiles de ácidos grasos de los alimentos y los gametos mediante cromatografía de gases, y se realizaron análisis estadísticos para determinar diferencias significativas. Resultados: Se observaron diferencias significativas en la abundancia (%) de ácidos grasos omega-3 (ω-3) y omega-6 (ω-6). La vía metabólica de (ω-3) fue más pronunciada en los gametos de los animales en su entorno natural y aquellos alimentados con los piensos experimentales. Por el contrario, la vía metabólica de (ω-6) tuvo menos relevancia en estos grupos. La relación (ω-3) /(ω-6) fue más alta en los gametos de los animales en su entorno natural. La disponibilidad de ácidos grasos en los alimentos permitió su bioacumulación en los gametos, con concentraciones iguales o superiores a las observadas en los animales en su entorno natural para ciertos ácidos grasos. Conclusiones: La incorporación de subproductos de camarones en los alimentos de acuicultura demostró ser una estrategia prometedora para la utilización de recursos y la generación de insumos orgánicos. La composición de ácidos grasos en los gametos de A. dufresnii fue influenciada por la dieta, destacando el potencial de los alimentos balanceados para mejorar la bioacumulación de ácidos grasos esenciales. Estos hallazgos brindan información valiosa para el desarrollo de prácticas sostenibles en acuicultura y la producción de productos marinos enriquecidos nutricionalmente.

Aphanomyces astaci in Mexico: A new haplotype from dwarf crayfish Cambarellus montezumae.

The crayfish plague is an emerging infectious disease caused by the pathogen Aphanomyces astaci (Oomycota), which is responsible for the decimation of Eurasian freshwater crayfish. This pathogen can coexist with the North American crayfish. These are chronic carriers of the disease as consequence of an immune response that can contain the growth of the pathogen without killing it. The origin of A. astaci locates in the southeastern United States and coincides with the origin of the family Cambaridae. This diverse family of decapods is distributed in North America from southern Canada to Honduras. However, only the native crayfish species from Canada and the USA have been examined for the presence of A. astaci. In this study, we describe for the first time the presence of A. astaci in Mexico in a population of the native species Cambarellus montezumae. By analyzing the small (rrnS) and large (rrnL) mitochondrial ribosomal regions, we showed the presence of two haplotypes of A. astaci within the same population (d1-haplotype and, a novel haplotype that was named, mex1-haplotype). The finding of A. astaci in Mexico confirms the occurrence of this pathogen within the range of the family Cambaridae. The individuals of C. montezumae appear to be chronic carriers of A. astaci, indicated by the lack of documented crayfish plague outbreaks in this population, similar to the pattern observed in other North American species. Thus, the results are of special concern to susceptible species of southern regions of America, i.e., Parastacidae. Therefore, this work emphasizes the need to better understand the distribution and genetic diversity of A. astaci within the distribution range of the natural carriers, i.e., North American species, especially the unexplored area of the family Cambaridae.

Oxidative stress and histopathological effects by microplastic beads, in the crayfish Procambarus clarkii, and fiddler crab Leptuca pugilator.

The present study was aimed at evaluating the in vivo effects of microplastics (MP), in terms of oxidative stress and histopathological effects, in two crustacean species: Procambarus clarkii and Leptuca pugilator. In addition, MP accumulation in the hepatopancreas (HP) of both species was also determined. Adults of both crayfish and crabs were exposed for one month to fluorescent polystyrene beads (size: 1 µm) at nominal concentrations of 1000 or 5000 particles/mL. During the exposure, animals were maintained under controlled feeding, aeration, temperature, and photoperiod conditions. At the end of the exposure, HP and hemolymph (HL) samples were harvested for analysis of oxidative damage and total antioxidant levels. Additionally, the presence of MPs in both tissues was confirmed. Significant differences with the control groups were observed in lipid peroxidation levels in HP in animals exposed to the lowest concentration in P. clarkii and to the highest concentration in L. pugilator. A marked increase in antioxidant levels was also observed in the HL at both concentrations in P. clarkii, and at the highest MPs concentration in L. pugilator. Moreover, several histopathological changes were detected in both gills and HP, including hypertrophied lamellae, lifting or collapse of gill epithelia, loss of normal shape of hepatopancreatic tubules, and epithelial atrophy in the HP tissue. We conclude that exposure to MP beads at selected concentrations results in oxidative damage, induces histopathological changes in gills and HP, and triggers an antioxidant response in two crustacean species.

Estudio de la calidad del agua en estanques e implementación de un protocolo de buenas prácticas acuícolas en la producción de camarón marino en Camaronera Eben Ezer, San Alejo, La Unión / Study of water quality in ponds and implementation of a Good Aquaculture Practices Protocol in the production of marine shrimp at the Camaronera Eben-Ezer, San Alejo, La Unión

La Escuela Especializada en Ingeniería ITCA FEPADE, MEGATEC La Unión, a través de la carrera Técnico en Manejo Integrado de Recursos Costero Marino con especialidad en Acuicultura y Pesquería, realizó esta investigación en asocio con Camaronera Eben Ezer. El proyecto tuvo como objetivo la caracterización de la calidad física, química y biológica del agua del Golfo de Fonseca y el Estero El Chapernal, previo al desarrollo de dos ciclos de cultivo de camarón marino Litopenaeus vannamei, durante el cultivo y antes de las descargas de agua a los efluentes receptores. La metodología se desarrolló en tres fases, de junio a diciembre de 2021. Fase de campo: toma de parámetros físicos, químicos y biológicos en 8 puntos de muestreos, Estero El Chapernal, Golfo de Fonseca y estanques de producción. Fase de laboratorio: se realizó sembrando muestras de agua en medios de cultivo para aislamiento, crecimiento e identificación de bacterias. Tercera fase: procesamiento de datos y análisis de resultados. Se identificaron tres tipos de bacterias: Psudomonas, Vibrios y Heterótrofas. En el primer ciclo de junio a agosto, éstas no reportaron crecimiento, esto pudo deberse a que la transición de la época seca a época lluviosa influyó en los factores físicos y químicos, como la temperatura y el pH. Durante el primer ciclo, la calidad del agua del Estero El Chapernal mantuvo estándares idóneos para el crecimiento del camarón. El segundo ciclo presentó un crecimiento exponencial de las bacterias Heterótrofas y Pseudomonas sobrepasando los límites de referencia establecidos en la normativa vigente. Como parte de la proyección social se efectuó transferencia de conocimientos y tecnología durante la producción, se establecieron mejoras para la operatividad y se les dotó de un protocolo de producción acuícola.

The Escuela Especializada en Ingeniería ITCA FEPADE, MEGATEC La Unión, through the major of Técnico en Manejo Integrado de Recursos Costero Marino con especialidad en Acuicultura y Pesquería, carried out this research in association with Camaronera Eben Ezer. The objective of the project was to characterize the physical, chemical and biological quality of water in Golfo de Fonseca and Estero El Chapernal, prior to the development of two culture cycles of the marine shrimp Litopenaeus vannamei, during the culture and before discharges of water to receiving effluents. The methodology was developed in three phases, from June to December 2021. Field phase: collection of physical, chemical and biological parameters at 8 demonstration points, Estero El Chapernal, Golfo de Fonseca and production ponds. Laboratory phase: it was carried out by planting water samples in culture media for isolation, growth and identification of bacteria. Third phase: data processing and analysis of results. Three types of bacteria were identified: Psudomonas, Vibrios and Heterotrophs. In the first cycle from June to August, growth was not reported, due to the fact that the transition from the dry season to the rainy season influenced in physical and chemical factors, such as temperature and pH. During the first cycle, the water quality of Estero El Chapernal maintained optimal standards for shrimp growth. The second cycle presented an exponential growth of Heterotrophic and Pseudomonas bacteria, exceeding the reference limits established in current regulations. As part of the social projection, knowledge and technology were transferred during production, they were profoundly improved for operability and they were provided with an aquaculture production protocol.

cDNA Characterization and Expression of Selenium-Dependent CqGPx3 Isoforms in the Crayfish Cherax quadricarinatus under High Temperature and Hypoxia.

Glutathione peroxidase 3 (GPx3) is the only extracellular selenoprotein (Sel) that enzymatically reduces H2O2 to H2O and O2. Two GPx3 (CqGPx3) cDNAs were characterized from crayfish Cherax quadricarinatus. The nerve cord CqGPx3a isoform encodes for a preprotein containing an N-terminal signal peptide of 32 amino acid residues, with the mature Sel region of 192 residues and a dispensable phosphorylation domain of 36 residues. In contrast, the pereiopods CqGPx3b codes for a precursor protein with 19 residues in the N-terminal signal peptide, then the mature 184 amino acid residues protein and finally a Pro-rich peptide of 42 residues. CqGPx3 are expressed in cerebral ganglia, pereiopods and nerve cord. CqGPx3a is expressed mainly in cerebral ganglia, antennulae and nerve cord, while CqGPx3b was detected mainly in pereiopods. CqGPx3a expression increases with high temperature and hypoxia; meanwhile, CqGPx3b is not affected. We report the presence and differential expression of GPx3 isoforms in crustacean tissues in normal conditions and under stress for high temperature and hypoxia. The two isoforms are tissue specific and condition specific, which could indicate an important role of CqGPx3a in the central nervous system and CqGPx3b in exposed tissues, both involved in different responses to environmental stressors.

Genomic scale data shows that Parastacus nicoleti encompasses more than one species of burrowing continental crayfishes and that lineage divergence occurred with and without gene flow.

Delimiting species is a challenge, especially in scenarios of diversification with gene flow and when species are now allopatric where reproductive isolation cannot be directly tested. Continental burrowing crayfishes of the genus Parastacus present a disjoint distribution in southern South America. One of the species is P. nicoleti, which lives in underground waters in swampy and wooded areas of southern Chile. A previous assessment based on mitochondrial DNA sequences suggest that the taxon may represent a species complex. Here, using thousands of nuclear genomic single-nucleotide polymorphisms obtained via RADSeq from 81 specimens collected at 27 localities throughout the distributional range of the species, we apply an integrative species delimitation approach to test species boundaries and to investigate some aspects of the speciation process. Our analyses corroborate previous results; a scenario that we favor suggests that the P. nicoleti encompasses seven distinct species. Additionally, demographic analyses show that the distinct species have followed distinct trajectories in size change during the last 17.5 million years and that speciation in this group occurred both in strict isolation as well as in the presence of gene flow.

The invasive crayfish Cherax quadricarinatus facing chlordecone in Martinique: Bioaccumulation and depuration study.

The redclaw crayfish, Cherax quadricarinatus, was introduced to Martinique Island for aquaculture purposes at the beginning of the 21st century, in an attempt to revitalize the freshwater crustacean aquaculture sector. Mainly due to its high economical value, it was intentionally released in the wild and was caught and sold by fishermen. Martinican rivers are polluted by chlordecone, considered as one of the worst Persistant Organic Pollutants (POP). Despite its dangerousness, it was used until 1993 in the French West Indies against a banana pest and was always found in the ecosystems. This study aimed to investigate the level of contamination in the muscle of crayfish caught in the wild, as well as the potential of bioconcentration and depuration in the C. quadricarinatus muscle. This study could allow us to quantify the risk for consumers but also, to evaluate a depuration process to reduce the risk related to its consumption. Using both in-vitro and in-situ experiments, results highlighted the importance of the chlordecone concentration in the water and the time of exposure to the pollutant. The bioconcentration seems to be very quick and continuous in crayfish muscle, as chlordecone can be detectable as early as 6 h of exposure, whatever the concentration tested. Finally, it appears that, even after 20 days of depuration in chlordecone-free water, chlordecone concentrations remained higher to the residual maximum limit (i.e. 20 ng/g wet weight), concluding that the decontamination of the muscle seems not very efficient, and the risk for the Martinican people could be serious.

Size distribution and sex ratio between populations of the artisanal harvested land crab Cardisoma guanhumi (Decapoda: Gecarcinidae), with the estimation of relative growth and size at sexual maturity in Puerto Rico / Distribución de tallas y proporción de sexos entre poblaciones del cangrejo de tierra cosechado artesanalmente Cardisoma guanhumi (Decapoda: Gecarcinidae), con la estimación del crecimiento relativo y tamaño en la madurez sexual en Puerto Rico

Abstract Introduction: Knowledge of growth patterns, sex ratio, and sexual maturity are of importance to exploited populations. The land crab Cardisoma guanhumi is an artisanal and subsistence exploited species in Puerto Rico. However, the growth patterns and sexual maturity of the local populations are not known. Objectives: This study has a double objective: (1) to compare the size and sex structure between populations and (2) to model the relative growth of structures related to reproduction to estimate the average size of morphometric sexual maturity (MSM) for both males and females. Methods: A total of 2 849 specimens were captured from nine dispersed populations on the island between 2001 and 2020. Carapace width (CW) was measured as an estimator of the absolute size of all individuals, together with the propodus length (PL) in males and the abdomen width in females (AW). Differences in length structure between sexes and populations were tested by applying goodness-of-fit tests based on Kernel Density Estimators (KDE). The relative growth pattern was modeled adjusting a spline from which the maximum of its second derivative was calculated as an estimator of the MSM, and bootstrapping was used to generate confidence intervals. Results: Differences were found in size structures, between sexes, and between sites. Our estimates of morphometric sexual maturity resulted in a sexual maturity size for males between 57.9 and 79.0 mm CW, while in females, morphological maturity occurs between 43.8 to 51.5 mm CW. Conclusions: We found inter-population differences in body size that can be attributed to differences in the history of changes of land use and the exploitation biased towards larger individuals, though differences in recruitment should also be considered. Current regulations in Puerto Rico protect female crabs but not to larger male crabs. The regulation establishes that crabs smaller than 64 mm carapace width cannot be captured, leaving immature male crabs over 64 mm CW unprotected. We suggest considering different size limits depending on the sex of the crab. This will allow the full range of sizes where sexual maturity is reached to be protected, increasing the likelihood of the population's size to increase.

Resumen Introducción: El conocimiento de los patrones de crecimiento, la proporción de sexos y la madurez sexual son de suma importancia para las poblaciones explotadas. El cangrejo terrestre Cardisoma guanhumi es una especie explotada artesanalmente y para subsistencia en Puerto Rico. Sin embargo, se desconocen los patrones de crecimiento y la madurez sexual para la isla. Objetivos: Este estudio tiene un doble objetivo: (1) comparar el tamaño y la estructura sexual entre las poblaciones y (2) modelar el crecimiento relativo de estructuras relacionadas con la reproducción (propodo en machos y abdomen en hembras) para estimar el tamaño promedio de madurez sexual morfométrica (MSM). Métodos: Se capturaron un total de 2 849 especímenes de nueve poblaciones dispersas en la isla, entre 2001 y 2020. Se midió el ancho del caparazón (AC) como un estimador del tamaño absoluto de todos los individuos, junto con el largo de propodo en los machos y el ancho del abdomen en las hembras. Las diferencias en la estructura de tallas entre sexos y poblaciones se probaron aplicando la prueba de bondad de ajuste basado en estimaciones de densidad kernel (KDE). El patrón de crecimiento relativo se modeló ajustando un spline a partir del cual se calculó el máximo de su segunda derivada como estimador del MSM y se utilizó bootstrapping para generar intervalos de confianza. Resultados: Se encontraron diferencias en las estructuras de tallas, entre sexos y poblaciones. Nuestras estimaciones de madurez sexual morfométrica dieron como resultado un tamaño de madurez sexual para los machos entre 57.9 y 79.0 mm AC, mientras que, en las hembras, la madurez morfométrica se produce entre 43.8 y 51.5 mm AC. Conclusiones: La diferencia en el tamaño de la población se puede atribuir a las diferencias entre poblaciones en el cambio de uso de la tierra y la presión de explotación asociada con el sesgo hacia la captura de individuos más grandes o el reclutamiento. Las regulaciones actuales en Puerto Rico brindan protección a las hembras, pero no a los machos más grandes. El reglamento establece que no se pueden capturar cangrejos de menos de 64 mm de AC dejando desprotegidos los cangrejos machos inmaduros de más de 64 mm AC. Sugerimos considerar diferentes límites de tamaño según el sexo del cangrejo. Esto permitirá proteger un rango más amplio de tamaños donde se alcanza la madurez sexual, aumentando la probabilidad de un incremento en el tamaño de la población.

Aerobic and anaerobic metabolic scaling in the burrowing freshwater crayfish Parastacus pugnax.

Metabolic scaling is a well-known biological pattern. Theoretical scaling exponents near 0.67 and 0.75 are the most widely accepted for aerobic metabolism, but little is known about the scaling of anaerobic metabolism. Furthermore, metabolic scaling has been mainly evaluated in organisms primarily relying on aerobic pathways. Here we evaluate both aerobic and anaerobic metabolic scaling in Parastacus pugnax, a burrowing freshwater crayfish endemic to Chile, which inhabits waters with low pO2 (~ 1 mg O2 L-1, measured in this study). We determined the metabolic rate, total oxidative capacity (Electron Transport System: ETS), critical oxygen tension (Pcrit) and muscular Lactate dehydrogenase (LHD) and Malate dehydrogenase (MDH) enzymatic activities (proxies of anaerobic metabolism) over a wide range of P. pugnax sizes (0.24-42.93 g wet mass). Aerobic metabolism scaled with crayfish size with an exponent of 0.78, remarkably similar to the 0.73 which scaled the ETS, the enzymatic complex behind respiration. Critical partial pressure of oxygen (Pcrit) was calculated as 15.6 ± 2.9 mmHg, showing that aerobic metabolism was efficiently maintained until ~ 10% air saturation. Below this threshold, P. pugnax switched to anaerobic metabolism, evidenced by a reduction in aerobic metabolism and ETS activity under chronic low oxygen conditions. None of the activities of MDH, LDH, their ratio (MDH/LDH), nor Pcrit scaled with crayfish size, indicating that these animals are equally adapted to hypoxic environments throughout their whole ontogeny. Given the particularities of its habitat, the information presented here is valuable for a proper management and successful conservation.

Evolving in islands of mud: old and structured hidden diversity in an endemic freshwater crayfish from the Chilean hotspot.

Parastacus is a genus of South American freshwater crayfishes disjunctively distributed in southern Chile, Northern Argentina, Uruguay and Southeastern Brazil. Parastacus pugnax is a Chilean endemic distributed along 700 km of latitude in central-southern Chile from the Pacific coast to the Andean piedmont, which is intensively captured for consumption for local communities. Considering the habitat (wet meadows) and natural history (primary burrower, non-migrant) of the species, we tested a hypothesis of highly structured genetic diversity using mtDNA of 465 specimens gathered at 56 localities across the species range. The crown age of P. pugnax was estimated at 38 Ma, predating the main Andean uplift. The genetic variation of P. pugnax is large and geographically structured. In some cases, genetic groups do not match basin limits, suggesting a previous to current dynamic of basin evolution. The uncovered intraspecific main lineages have different demographic histories. A latitudinal cline in past effective population size reduction suggests environmental singularities with a glacial effect in the southern populations. We suggest adding morphologic and more genetic data in order to assess species limits. Our results contribute to improve future conservation actions for this taxon, providing basic information to delimit conservation units.

Áreas en riesgo de invasión por Procambarus clarkii (Decapoda: Cambaridae) un cangrejo de río introducido en Colombia / Areas at risk of invasion by Procambarus clarkii (Decapoda: Cambaridae) a crayfish introduced in Colombia

Introducción: El proceso de invasión biológica es una de las mayores amenazas a la biodiversidad y ecosistemas, con actuales y potenciales impactos en la salud pública y conservación. Procambarus clarkii es un crustáceo decápodo, originario del sur de Estados Unidos y noreste de México, que puede adaptarse a diferentes condiciones ambientales debido a su plasticidad ecológica. Objetivo: Se caracterizó el nicho ecológico fundamental existente de Procambarus clarkii con el fin de predecir las áreas con idoneidad ambiental para el potencial establecimiento de la especie en Suramérica y Colombia. Métodos: Usamos modelos de nichos ecológicos calibrados en el área nativa, elaborados con el algoritmo Maxent, basados en datos de presencia extraídos de GBIF y variables hidroclimáticas de ecosistemas acuáticos a una resolución de 1 km2. Resultados: En Suramérica el modelo indicó un potencial geográfico amplio de invasión mostrando áreas ambientalmente idóneas para la presencia y expansión hacia Colombia, Venezuela, Perú, Ecuador, Brasil, Guyana, Surinam, Bolivia, Uruguay, Argentina, Paraguay y Chile. En Colombia, el modelo predijo que las áreas idóneas se ubican principalmente en el norte y oriente, incluidos diversos ecosistemas, como: bosques tropicales, bosques basales, bosques riparios y sabanas. La especie tiene una alta posibilidad de expandirse hacia áreas de distribución de latitudes bajas, ocupando zonas hacia la parte norte de la región Caribe colombiana, en departamentos de Magdalena, Cesar, Córdoba y Atlántico. También se predijo áreas con idoneidad ambiental en el oriente de Colombia, hacia la extensión de la planicie oriental de la Orinoquia, una región de baja altitud en: Arauca, Casanare, Meta y Vichada. Conclusión: En este estudio se aplican modelos de nichos ecológicos, que puede ser de interés en la planeación de estrategias o la creación de planes de manejo, como sistemas de alerta temprana para evitar el establecimiento de esta especie.

Introduction: Biological invasions are a major threat to biodiversity and ecosystems, with current and potential impacts on public health and conservation. Procambarus clarkii is a decapod crustacean, native to the Southern United States and Northeastern Mexico, which may adapt to different environmental conditions due to its ecological plasticity. Objective: The existing fundamental ecological niche of Procambarus clarkii was characterized in order to predict areas with environmental suitability for the potential establishment of the species in South America and Colombia. Methods: We used models of calibrated ecological niches in the native area, elaborated with the Maxent algorithm, based on occurrence data extracted from GBIF and hydroclimatic variables of aquatic ecosystems at a resolution of 1 km2. Results: The model indicated a wide geographic area for invasion potential in South America, predicting environmentally suitable areas for the presence and expansion towards Colombia, Venezuela, Peru, Ecuador, Brazil, Guyana, Surinam, Bolivia, Uruguay, Argentina, Paraguay and Chile. In Colombia, suitable areas predicted by the model are located mainly in the North and East of the country, spanning a diversity of ecosystems, such as tropical forests, basal forests, riparian forests and savannas. The species has a high possibility of expanding into low latitude distribution areas, occupying areas towards the Northern part of the Colombian Caribbean region, in the departments of Magdalena, Cesar, Cordoba and Atlántico. Suitability areas environmental were also predicted in Eastern Colombia, towards the extension of the Eastern plain of the Orinoquia, a low altitude region in Arauca, Casanare, Meta and Vichada. Conclusion: This study applies ecological niche models, which may be of interest in the planning of strategies or the creation of management plans, such as early warning systems that prevent the establishment of this species.

Assessment of interspecific interactions between the invasive red-claw crayfish (Cherax quadricarinatus) and the Mozambique tilapia (Oreochromis mossambicus) / Avaliação das interações interespecíficas entre o lagostim invasivo de garra vermelha (Cherax quadricarinatus) e a tilapia de Moçambique (Oreochromis mossambicus)

The Australian red-claw crayfish, Cherax quadricarinatus, has been introduced for aquaculture purposes worldwide and consequently colonized natural environments, where it might cause ecosystem services losses or adversely affect native species and the local environment. This species was first found in Pequenos Libombos Reservoir in Maputo Province, Southern Mozambique in the late 2009 and is linked to reduction in tilapia fisheries. This study, conducted in 2015 under controlled conditions, aimed to assess the interspecific relationships between the Mozambique tilapia (Oreochromis mossambicus) and the alien crayfish. For both species, no significant differences in growth and survival rates were found between animals reared in the presence versus the absence of heterospecifics, indicating no direct deleterious interspecific effects. Behavioural observations revealed that fish and crayfish competed for shelter and food. Both species reduced the foraging in the presence of heterospecifics during feeding period, in the daytime. Crayfish seemed to have an advantage in competition for shelter, suggesting that they may interfere with tilapia sheltering activity and make tilapia vulnerable to predators in natural habitats.(AU)

O lagostim australiano, Cherax quadricarinatus, tem sido introduzido para fins de aquicultura em todo o mundo e, consequentemente, colonizado ambientes naturais, onde pode coexistir com a biota local sem causar perdas nos serviços dos ecossistemas ou afetar negativamente as espécies nativas e o meio ambiente local. Esta espécie foi encontrada pela primeira vez na Albufeira dos Pequenos Libombos, na Província de Maputo, sul de Moçambique, no final de 2009 e foi relacionada à redução da pesca de tilápias registrada na mesma localidade. Este estudo, conduzido em 2015 sob condições controladas, teve como objetivo avaliar as relações interespecíficas entre a tilápia de Moçambique (Oreochromis mossambicus) e o lagostim invasor (C. quadricarinatus). Para ambas espécies, não foram encontradas diferenças significativas nas taxas de crescimento e sobrevivência entre animais criados na presença versus ausência de heteroespecíficos, indicando ausência de efeitos deletérios interespecíficos diretos. Observações comportamentais revelaram que peixes e lagostins competiam por abrigo e comida. Ambas espécies reduziram o forrageamento na presença de heteroespecíficos durante o período de alimentação, no período diurno. O lagostim parece ter uma vantagem na competição por abrigos, sugerindo que, em habitats naturais eles podem provocar o deslocamento de tilápias de seus abrigos, e deste modo tornar as tilápias vulneráveis aos predadores.(AU)

Assessment of interspecific interactions between the invasive red-claw crayfish (Cherax quadricarinatus) and the Mozambique tilapia (Oreochromis mossambicus) / Avaliação das interações interespecíficas entre o lagostim invasivo de garra vermelha (Cherax quadricarinatus) e a tilapia de Moçambique (Oreochromis mossambicus)

Abstract The Australian red-claw crayfish, Cherax quadricarinatus, has been introduced for aquaculture purposes worldwide and consequently colonized natural environments, where it might cause ecosystem services losses or adversely affect native species and the local environment. This species was first found in Pequenos Libombos Reservoir in Maputo Province, Southern Mozambique in the late 2009 and is linked to reduction in tilapia fisheries. This study, conducted in 2015 under controlled conditions, aimed to assess the interspecific relationships between the Mozambique tilapia (Oreochromis mossambicus) and the alien crayfish. For both species, no significant differences in growth and survival rates were found between animals reared in the presence versus the absence of heterospecifics, indicating no direct deleterious interspecific effects. Behavioural observations revealed that fish and crayfish competed for shelter and food. Both species reduced the foraging in the presence of heterospecifics during feeding period, in the daytime. Crayfish seemed to have an advantage in competition for shelter, suggesting that they may interfere with tilapia sheltering activity and make tilapia vulnerable to predators in natural habitats.

Resumo O lagostim australiano, Cherax quadricarinatus, tem sido introduzido para fins de aquicultura em todo o mundo e, consequentemente, colonizado ambientes naturais, onde pode coexistir com a biota local sem causar perdas nos serviços dos ecossistemas ou afetar negativamente as espécies nativas e o meio ambiente local. Esta espécie foi encontrada pela primeira vez na Albufeira dos Pequenos Libombos, na Província de Maputo, sul de Moçambique, no final de 2009 e foi relacionada à redução da pesca de tilápias registrada na mesma localidade. Este estudo, conduzido em 2015 sob condições controladas, teve como objetivo avaliar as relações interespecíficas entre a tilápia de Moçambique (Oreochromis mossambicus) e o lagostim invasor (C. quadricarinatus). Para ambas espécies, não foram encontradas diferenças significativas nas taxas de crescimento e sobrevivência entre animais criados na presença versus ausência de heteroespecíficos, indicando ausência de efeitos deletérios interespecíficos diretos. Observações comportamentais revelaram que peixes e lagostins competiam por abrigo e comida. Ambas espécies reduziram o forrageamento na presença de heteroespecíficos durante o período de alimentação, no período diurno. O lagostim parece ter uma vantagem na competição por abrigos, sugerindo que, em habitats naturais eles podem provocar o deslocamento de tilápias de seus abrigos, e deste modo tornar as tilápias vulneráveis aos predadores.

Analysis of the population structure and genetic diversity of the red swamp crayfish (Procambarus clarkii) in China using SSR markers

BACKGROUND: Procambarus clarkii produces high-quality, delicious meat that is high in protein, low in fat, and rich in calcium and phosphorus. It has become an important aquatic resource in China. Our objectives are (i) to analyze the level of genetic diversity of P. clarkii populations; (ii) to explore the genetic differentiation (Gst); and (iii) to propose appropriate strategies for the conservation. RESULTS: In this study, Shannon's index (I) and Nei's gene diversity index (H) for P. clarkii were high (I = 0.3462 and H = 0.2325 on average and I = 0.6264, H = 0.4377 at the species level) based on the SSR markers. The expected heterozygosity value of 17 microsatellite loci in 25 crayfish populations was 0.9317, the observed heterozygosity value was 0.9121, and the observed number of alleles per locus was 2.000; and the effective number of alleles per locus was 1.8075. Among the P. clarkii populations, the inbreeding coefficient within populations (Fis) was 0.2315, overall inbreeding coefficient (Fit) was 0.4438, genetic differentiation coefficient among populations (Fst) was 0.3145 and gene differentiation (Gst) was 0.4785 based on SSR analyses. The cluster analysis results obtained by unweighted pair-group method with arithmetic mean (UPGMA) analysis, principal coordinate analysis (PCoA) and STRUCTURE analysis were similar. A mantel test showed that the isolation-by-distance pattern was not significant. CONCLUSIONS: The high Gst among P. clarkii populations is attributed to genetic drift and geographic isolation. The results indicated that more P. clarkii populations should be collected when formulating conservation and aquaculture strategies.

Correlation profile of the heavy metal distribution in the Pontastacus leptodactylus tissues.

Arsenic (As), copper (Cu), nickel (Ni), zinc (Zn), manganese (Mn), lead (Pb), iron (Fe), aluminum (Al), cadmium (Cd) and chromium (Cr) accumulations were evaluated in the gills, hepatopancreas, exoskeleton, and muscles of Pontastacus leptodactylus (crayfish) (Eschscholtz, 1823). The highest metal accumulation was observed in the gills and hepatopancreas. It was detected a negative correlation between Cr-Pb, As-Cr in the muscle tissue. All other metals were displayed positive correlations with each other in the gills, hepatopancreas, and exoskeleton. Strong positive correlations were determined between Ni-Cd and As-Pb in the gills, Ni-Cd, As-Pb and Al-Zn in the hepatopancreas, Zn-Mn and Fe-Cu in the exoskeleton, Ni-Cd, As-Pb, Zn-Mn, Al-Mn, Fe-Cu and Al-Zn in the muscle (r= 1.000). PCA and cluster analysis generally were supported the correlations observed. The correlations between divalent metals may be expressed by the role of divalent metal transporter 1 (DMT1) in the gathering of these metals.

A Balancing Act: The Immune System Supports Neurodegeneration and Neurogenesis.

Decapod crustaceans, like mammals, retain the ability to make new neurons throughout life. In mammals, immune cells are closely associated with stem cells that generate adult-born neurons. In crayfish, evidence suggests that immune cells (hemocytes) originating in the immune system travel to neurogenic regions and transform into neural progenitor cells. This nontraditional immune activity takes place continuously under normal physiological conditions, but little is known under pathological conditions (neurodegeneration). In this study, the immune system and its relationship with neurogenesis were investigated during neurodegeneration (unilateral antennular ablation) in adult crayfish. Our experiments show that after ablation (1) Proliferating cells decrease in neurogenic areas of the adult crayfish brain; (2) The immune response, but not neurogenesis, is ablation-side dependent; (3) Inducible nitric oxide synthase (iNOS) plays a crucial role in the neurogenic niche containing neural progenitors during the immune response; (4) Brain areas targeted by antennular projections respond acutely (15 min) to the lesion, increasing the number of local immune cells; (5) Immune cells are recruited to the area surrounding the ipsilateral neurogenic niche; and (6) The vasculature in the niche responds acutely by dilation and possibly also neovascularization. We conclude that immune cells are important in both neurodegeneration and neurogenesis by contributing in physiological conditions to the maintenance of the number of neural precursor cells in the neurogenic niche (neurogenesis), and in pathological conditions (neurodegeneration) by coordinating NO release and vascular responses associated with the neurogenic niche. Our data suggest that neural damage and recovery participate in a balance between these competing immune cell roles.

Assessment of interspecific interactions between the invasive red-claw crayfish (Cherax quadricarinatus) and the Mozambique tilapia (Oreochromis mossambicus).

The Australian red-claw crayfish, Cherax quadricarinatus, has been introduced for aquaculture purposes worldwide and consequently colonized natural environments, where it might cause ecosystem services losses or adversely affect native species and the local environment. This species was first found in Pequenos Libombos Reservoir in Maputo Province, Southern Mozambique in the late 2009 and is linked to reduction in tilapia fisheries. This study, conducted in 2015 under controlled conditions, aimed to assess the interspecific relationships between the Mozambique tilapia (Oreochromis mossambicus) and the alien crayfish. For both species, no significant differences in growth and survival rates were found between animals reared in the presence versus the absence of heterospecifics, indicating no direct deleterious interspecific effects. Behavioural observations revealed that fish and crayfish competed for shelter and food. Both species reduced the foraging in the presence of heterospecifics during feeding period, in the daytime. Crayfish seemed to have an advantage in competition for shelter, suggesting that they may interfere with tilapia sheltering activity and make tilapia vulnerable to predators in natural habitats.