Evidence of the Existence of Site-Specific Female Contact Pheromones Involved in the Sexual Interaction Behavior of the Pacific Whiteleg Shrimp Penaeus vannamei.

Although the presence of female contact sex pheromones in P. vannamei has been hypothesized, to date its existence has not been proven. To gather more evidence of their existence, cuticular liposoluble extracts were obtained from the following samples of adult females to be used as the experimental treatments: (1) ventral exoskeleton of immature female (VI), (2) dorsolateral exoskeleton of immature female (DI), (3) ventral exoskeleton of mature female (VM), and (4) dorsolateral exoskeleton of mature female (DM). Polyvinyl chloride tubes (artificial females; AF) were coated with each extract and the behavior displayed by sexually mature males in contact with the AF was recorded and classified as follows: 0 = no response; 1 = contact; 2 = pushing; and 3 = prolonged contact (≥10 s). To test the hypothesis that the extracts collected from the ventral portion of the abdomen exoskeleton have a higher effect on the behavior of males than the extracts collected from the dorsolateral portion of the abdomen exoskeleton, the experiment was divided into two bioassays: Bioassay I (VI vs. DI) and Bioassay II (VM vs. DM). In each bioassay, all experimental treatments were significantly different (p > 0.05) from the CTL group (AF coated with hexane). Notably, the pushing behavior was significantly higher (p < 0.05) in the VI treatment compared to the CTL and DI treatment. These results provide evidence of the existence of contact female sex pheromones with sexual recognition function located primarily in the ventral portion of the abdomen exoskeleton of P. vannamei.

Regulation of early spermatogenesis in the giant prawn Macrobrachium rosenbergii by a GCL homolog†.

The germ cell-less gene is crucial for gonad development in various organisms. Early interventions in its expression suggested a regulatory role at the mitotic stages of spermatogenesis, and its early knockout resulted in complete sterility in Drosophila. Genomic and transcriptomic data available for the catadromous giant prawn Macrobrachium rosenbergii enabled the identification of a germ cell-less homolog for this species, which we termed MroGCL (mRNA accession number OQ533056). An open reading frame containing 494 amino acids and a typical evolutionarily conserved BTB/POZ domain suggests possible protein-protein interaction functions in keeping with the Drosophila germ cell-less protein. Genomic mapping of MroGCL showed a full length of 120 896 bases. Analysis of the temporal expression of MroGCL showed constant expression in early prawn embryonic and larval stages, but a significant increase 10 days after metamorphosis when crucial sexual differentiation processes occur in prawns. In adult animals, high expression was detected in the gonads compared to the somatic tissues. RNAi-based knock-down experiments showed that both the silenced and control groups reached advanced spermatogenic stages, but that there was a significant decrease in the yield of spermatozoa in about half of the silenced animals. This finding supports our hypothesis that MroGCL is crucial for mitosis during early stage spermatogenesis. In conclusion, this study contributes to the understanding of crustacean gonad development and provides a stepping stone in the development of environmentally valuable sterile crustacean populations.

IDENTIFICACIÓN MOLECULAR Y DISTRIBUCIÓN POTENCIAL DEL ANFÍPODO TERRESTRE Talitroides topitotum (Crustacea: Amphipoda: Talitridae) EN COSTA RICA / Molecular Identification and Potential Distribution of Terrestrial Amphipod Talitroides topitotum (Crustacea: Amphipoda: Talitridae) in Costa Rica

RESUMEN El anfípodo terrestre, Talitroides topitotum, es un talítrido distribuido mundialmente en regiones subtropicales y templadas, con un amplio rango de distribución altitudinal, temperatura y humedad. Se colectaron y procesaron especímenes desde el año 2012 al 2016, mediante remoción-filtración de sustratos húmedos. Se identificaron taxonómicamente por características fenotípicas diagnósticas, se determinó su estado de desarrollo y se separaron por sexo. Se extrajo ADN de anfípodos completos, seguido de una PCR de los genes citocromo oxidasa subunidad 1 y del ARN ribosomal de la subunidad 16S. Se obtuvo un árbol filogenético por máxima verosimilitud con un modelo GTR-GAMMA. El análisis de la distribución potencial de T. topitotum se estimó utilizando 19 variables bioclimáticas. En este estudio, se amplía la distribución previamente reportada y en altitudes entre los 1900 a 595 m s.n.m. Se analizaron 39 localidades, en las cuales: 1) Hay presencia de T. topitotum, 2) no hubo presencia de anfípodos terrestres, 3) no hubo presencia de Talitroides sp., pero sí de un anfípodo nativo. La abundancia proporcional de T. topitotum se inclina hacia las hembras adultas, una proporción alta de juveniles y no se detectaron individuos machos. El análisis bioinformático determinó el posicionamiento taxonómico de la especie T. topitotum dentro del agrupamiento de anfípodos terrestres, además, la especie exógena diverge de Cerrorchestia hyloraina demostrando una separación filogenética entre especies, las cuales pueden estar compartiendo hábitats. T. topitotum, según el modelo de máxima entropía, posee una alta capacidad de dispersión y estaría siendo favorecida, en cuanto a su asentamiento y propagación, por elementos climáticos como temperatura, precipitación y humedad, y factores como la altitud. Nuestros hallazgos son relevantes para la toma de decisiones de manejo y monitoreo del desplazamiento de especies nativas de anfípodos terrestres en la región.

ABSTRACT The land-hopper, Talitroides topitotum, is a talitrid amphipod distributed worldwide in subtropical and template regions, with a wide range of altitudinal distribution, temperature and humidity. Specimens were collected and processed since 2012 until 2016, by collection-filtration of wet substrates. Specimens were taxonomically identified using diagnostic phenotypic characteristics, and the developmental stage and sex were recorded. DNA was extracted from whole amphipods, followed by PCR of cytochrome oxidase subunit 1 and ribosomal RNA subunit 16S genes. Partial genetic sequences were obtained and a maximum-likelihood phylogenetic tree was calculated based on a GTR-GAMMA model. The analysis of potential distribution of T. topitotum was estimated using 19 bioclimatic variables. This study extends the previously reported distribution and elevations between 1900 and 595 m a.s.l. Thirty-nine localities were analyzed, where the following categories were registered: 1) T. topitotum is present, 2) terrestrial amphipods are not present, 3) T. topitotum is not present, but the native amphipod is present. The relative abundance of T. topitotum corresponds to adult females, a high proportion ofjuveniles and no males were collected. The bioinformatic analysis established the taxonomic position of T. topitotum within a group of terrestrial amphipods; moreover, the invasive species diverges of Cerrorchestia hyloraina, demonstrating the phylogenetic separation between these species that could be sharing habitats. Based on the model of maximum entropy, T. topitotum shows a high dispersion capacity and its establishment and propagation are been improved by climatic elements such as temperature, precipitation, humidity, and elevation. Our findings are relevant for management policies and monitoring the distribution of native species of terrestrial amphipods in the region.