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Poster.-- XVIII Congreso Nacional de Acuicultura “Acuicultura: mares y ríos de oportunidades”, Cádiz, del 21 al 24 de noviembre de 2022, [EN] The common octopus, Octopus vulgaris, is a species with a high potential for aquaculture diversification. Recently, for the first time, it has been possible to obtain specimens up to the pre-settlement stage, with high levels of growth and survival, allowing the investigation of different biological processes. DNA methylation is an epigenetic process capable of controlling gene transcription, facilitating differential expression depending on environmental conditions. In the present work, RRBS sequencing and comparative study of the methylome of O. vulgaris paralarvae at different stages of development and subjected to different diet and temperature conditions has been carried out in order to assess how environmental conditions can vary the methylation profile of these organisms in culture conditions. The results obtained after the comparative analyses show the necessity of demethylation for the progress of their correct development. In addition, certain environmental conditions, such as low temperature or optimal diet, seem to facilitate demethylation. The transcriptional silencing or activation by methylation, together with increased annotation of the genes involved, will allow the identification of marker genes. The results of the present work will help to better understand the requirements for optimal development of this species, contributing to improve the transfer of the culture to industrial scale, [ES] El pulpo común, Octopus vulgaris, es una especie con un elevado potencial para la diversificación de la acuicultura. Recientemente se han obtenido por primera vez ejemplares hasta la fase de pre-asentamiento, con altos niveles de crecimiento y supervivencia, permitiendo investigar distintos procesos biológicos. La metilación del ADN es un proceso epigenético capaz de controlar la transcripción de los genes, facilitando una expresión diferencial en función de las condiciones del medio. En el presente trabajo se ha llevado a cabo la secuenciación por el método RRBS y estudio comparado del metiloma de paralarvas de O. vulgaris en diferentes estadios de desarrollo y sometidas a diferentes condiciones de dieta y temperatura con objeto de valorar cómo las condiciones ambientales pueden hacer variar su perfil de metilación en el cultivo. Los resultados obtenidos muestranla necesidad de la desmetilación para el avance de su correcto desarrollo. Además, determinadas condiciones ambientales, como la baja temperatura o una dieta óptima parecen facilitar dicha desmetilación. El silenciamiento o activación transcripcional mediante metilación, junto con el incremento de la anotación de los genes implicados, permitirán la identificación de genes marcadores. Los resultados obtenidos ayudarán a comprender mejor los requerimientos para un desarrollo óptimo de esta especie, contribuyendo a mejorar la transferencia del cultivo a escala industrial, Proyecto OCTOMICS (AGL-2017-89475-C2-1-R) financiado por el Ministerio de Ciencia e Innovación y por FEDER “Una manera de hacer Europa”

Poster.-- XVIII Congreso Nacional de Acuicultura “Acuicultura: mares y ríos de oportunidades”, Cádiz, del 21 al 24 de noviembre de 2022, [EN] The common octopus (Octopus vulgaris) is a species of great commercial interest and an excellent candidate for aquaculture diversification. Besides the administration of an adequate diet, culture conditions influence the maintenance of their health and welfare. The use of food supplements such as natural bioactive supplements administered with the diet has multiple beneficial effects. In this study we evaluated the effect of olive oil (HYT) and cumin oil (BCO) supplements on the expression of genes related to oxidative stress, immune function and pathogen recognition in cultured octopus paralarvae. The results show an increase in the expression of genes related to antioxidant function (cat, sod and gpx7) in paralarvae fed on HYT-supplemented Artemia. Among the genes related to immune function and pathogen recognition (gilt, c3, myd88, lys, tlr13 and pgrp), generally higher expression of these genes was detected after HYT administration, although lys, tlr13 and myd88 were also overexpressed in paralarvae fed on Artemia supplemented with BCO. In conclusion, the transcription of the genes analyzed was activated to a greater extent in the early life stages and after HYT administration and to a lesser extent with BCO. These bioactives could confer some kind of protection to octopus paralarvae in their early stages of development in culture, [ES] El pulpo común (Octopus vulgaris) es una especie de gran interés comercial y un excelente candidato para la diversificación de la acuicultura. Además de la administración de una dieta adecuada, las condiciones de cultivo influyen en el mantenimiento de su salud y bienestar. El uso de suplementos alimenticios como los complementos bioactivos naturales administrados con la dieta presenta múltiples efectos beneficiosos. En este trabajo se evaluó el efecto de suplementos derivados del aceite de oliva (HYT) y aceite de comino (BCO) en la expresión de genes relacionados con el estrés oxidativo, función inmune y reconocimiento de patógenos en paralarvas de pulpo en cultivo. Los resultados muestran un incremento en la expresión de genes relacionados con función antioxidante (cat, sod y gpx7) en paralarvas alimentadas con Artemia suplementada con HYT. Entre los genes relacionados con función inmune y reconocimiento de patógenos (gilt, c3, myd88, lys, tlr13 y pgrp), en general se detectó mayor expresión de estos genes tras la administración de HYT, aunque lys, tlr13 y MyD88 presentaron también sobreexpresión en paralarvas alimentadas con Artemia suplementada con BCO. En conclusión, la transcripción de los genes analizados se activó en mayor medida en los primeros estadios de vida y tras la administración de HYT y en menor medida con BCO. Dichos bioactivos podrían conferir algún tipo de protección a las paralarvas de pulpo en sus primeras etapas de desarrollo en cultivo, Proyecto OCTOMICS (AGL-2017-89475-C2-1-R) y proyecto IMMUNOCTOPUS (PID2020-119906GB-I00) financiados por el Ministerio de Ciencia e Innovación y por FEDER “Una manera de hacer Europa”.

Póster presentado en CephRes2020 Virtual Event Biology and Life History of Cephalopods, celebrado de manera virtual del 16 al 21 de septiembre de 2021., The common octopus (Octopus vulgaris) is an excellent candidate for aquaculture diversification. It is known that temperature has a great effect on early life stages of cephalopods, drivng the development of embryos, yolk absorption rate and growth rate of paralarvae, which increases with the temperature water. In this study, the impact of temperature incubation on octopus¿ embryos EMB (stage XV) and paralarvae after hatching (PL0) were investigated using the proteomic SWATH¿MS approach to identify proteins or protein profiles as biomarkers to evaluate the welfare in early stages of O. vulgaris. Two temperatures of incubation were compared: high (21 ºC, commonly used in paralarvae aquaculture) and low (16 ºC, as representative of natural conditions). The proteomic analysis quantified a total of 2352 proteins. For EMB, 24 proteins showed significant differential expression between high (H) and low (L) temperature, whereas in PL0 a total of 127 proteins presented significant differences. PCA analysis of all quantified proteins showed a clear separation between development stages (EMB and PL0) and temperature. The up-regulated proteins by high temperature in EMB (11 up) were mainly related with signaling pathways (signal transduction) scavenging and binding activity (metal, ions, tubulin and chitin). Whereas in PL0, the up-regulated proteins (64 up) by high temperature were involved mainly in proteolysis (hydrolysis activity related with protein metabolism), carbohydrate metabolism, nervous system development and neurotransmitter transport, lipid metabolic process, detoxification, extracellular organization and binding (chitin, metals and ATP). These results allow defining characteristic proteomes for EMB and PL0 that are involved in different biological processes associated to temperature., This research was supported by “Ministerio de Ciencia e Innovación through the “OCTOMICS Project” (AGL-2017-89475-C2-1-R).

Comunicación presentada en CephRes2020 Virtual Event Biology and Life History of Cephalopods, celebrado de manera virtual del 16 al 21 de septiembre de 2020., The common octopus Octopus vulgaris has been proposed as a good candidate for aquaculture operations due to its fast growth and short life cycle. Settlement of animals has been the main bottleneck to reach the adult stage under captivity. However, recent achievements have allowed the obtaining of samples to decipher the molecular basis that regulates the intricate early stages in this species. In this study, O. vulgaris samples at different stages were sequenced by RNA-seq for a massive de-novo transcriptomic analysis following the TRINITY protocol. Four main comparisons were carried out considering a mix of different embryonic stages and paralarvae at 0, 10, 20 and 40 days post-hatching (dph). A total of 2300 up-regulated transcripts were found in newly hatched paralarvae (0 dph) compared to embryo whereas the number of overexpressed transcripts reached 2336, 3013 and 2580 in 10, 20 and 40 dph paralarvae, respectively, compared to newly hatched animals. Gene ontology enrichment analysis revealed common terms at all ages such as chitin metabolism and peptidase related activities. Terms related to proteolysis and catabolism were significantly more represented in 0, 10 and 20 dph paralarvae whereas mobilization of proteins into the nucleus was more represented at 40 dph. Neural development and response to stimulus and activation of immune response were also differently represented attending to the age of the animals. Altogether these results represent an overview of the different biological functions at paralarvae stage and will help to decipher at what age relevant functions are activated.

Coastal protection, nutrient cycling, erosion control, water purification, and carbon sequestration are ecosystem services provided by salt marshes. Additionally, the salt ponds offers coastal breeding and nursery habitat to fishes and invertebrates providing abundant amphipods potentially useful as a resource in aquaculture. Fishmeal and fish oil are necessary food resources to support aquaculture of carnivorous species due to their omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA). Their dependence on limited fisheries constrains growth of aquaculture and the need for n-3 LC-PUFA recommends the development of more sustainable food sources. Due to their extensive feeding on detritus and primary producers, amphipods appear as potential and high quality alternative feed resource for aquaculture. Hence, a nutritional study was carried out for main amphipods species -Microdeutopus gryllotalpa, Monocorophium acherusicum, Gammarus insensibilis, Melita palmata and Cymadusa filosa- at South Spain aquaculture terrestrial ponds . These species showed a high protein content (up to 40%), high n-3 PUFA and phospholipid levels, and high levels of phophatidylcholine (PC), phosphatidylethanolamine (PE) and triacylglycerols(TAG), the latter significantly high for M. acherusicum . M. gryllotalpa and M. acherusicum showed the highest proportion of lipids (19.15% and 18.35%, respectively). Isoleucine, glycine and alanine were the dominant amino acids in all species. In addition, amphipods collected from ponds showed low levels of heavy metals. Furthermore, the biochemical profile of the five amphipods have been compared with other studied alternatives preys. Therefore, amphipods of ponds are good candidates to be used as feed, and are proposed as a new sustainable economic resource to use in aquaculture. G. insensibilis may be the best to be intensively cultured as an alternative feed resource because shows: 1) adequate PUFAS n-3 and PL composition; 2) high levels of glycine, alanine, tyrosine, isoleucine and lysine; 3) high natural densities; 4) large body size ( ≥ 1 cm) and 5) high concentration of Calcium. Moreover, a combined culture of amphipods and fishes in these marsh ponds seems a promising and environmentally sustainable way to develop Integrate Multi-Trophic Aquaculture (IMTA) in these ecosystems.

The physiological responses of Senegalese sole to a sudden salinity change were investigated. The fish were first acclimated to an initial salinity of 37.5 ppt for 4 h. Then, one group was subjected to increased salinity (55 ppt) while another group was subjected to decreased salinity (5 ppt). The third group (control group) remained at 37.5 ppt. We measured the oxygen consumption rate, osmoregulatory (plasma osmolality, gill and kidney Na+,K+-ATPase activities) and stress (plasma cortisol and metabolites) parameters 0.5 and 3 h after transfer. Oxygen consumption at both salinities was higher than for the control at both sampling times. Gill Na+,K+-ATPase activity was significantly higher for the 55 ppt salinity at 0.5 h. Plasma osmolality decreased in the fish exposed to 5 ppt at the two sampling times but no changes were detected for high salinities. Plasma cortisol levels significantly increased at both salinities, although these values declined in the low-salinity group 3 h after transfer. Plasma glucose at 5 ppt salinity did not vary significantly at 0.5 h but decreased at 3 h, while lactate increased for both treatments at the first sampling time and returned to the control levels at 3 h. Overall, the physiological response of S. senegalensis was immediate and involved a rise in oxygen consumption and plasma cortisol values as well as greater metabolite mobilization at both salinities.

36 p., 7 tables, 5 figures and bibliography, Effects of two binders (gelatine and alginate), were tested on growth, survival, partial energy balance and lipid composition of mantle and digestive gland of Octopus vulgaris. Octopus average weight was of 713.0±127.9 g at the start of the experiment. The three diets tested (10 octopuses per diet) were: 1) CON, (Loligo gahi) as a control diet, 2) GEL, composed of squid paste (L. gahi) (30%), fish hydrolyse CPSP® (10%), and high quality fish meal (50%), agglutinated with 10% of gelatine and 3) ALG, composed of squid paste (L. gahi) (30%), fish hydrolyse CPSP® (10%), and high quality fish meal (50%), all agglutinated with 10% of alginate. Growth rates were 1.4±0.2, 02±0.2 and -0.2±0.2 % bw d-1, for octopuses fed CON, GEL and ALG diets, respectively, and were higher when fed CON, compared to the two artificial diets. The GEL diet promoted higher growth and none of the artificial diets tested promoted significant growth. The digestive gland of octopuses had higher concentrations of fatty acids than the mantle. Fatty acid composition of the mantle was similar between CON and the two artificial diets. Digestive gland of animals fed CON had higher concentrations of fatty acids, than those fed the artificial diets., The authors wish to thank the “Plan Nacional de pulpo” – JACUMAR – Project “Engorde de pulpo, Octopus vulgaris.” 2007/2009, and the project CONACYT –Basico 50118 of C.R. from the Mexican government, for the funding for this research. Sandra García whishes to thank the “Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria” (INIA) for the Pre-doctoral grant nº 47 (BOE nº 308 26/12/2006).

4 páginas, 2 figuras. XI Congreso Nacional de Acuicultura (Vigo, 24-28 septiembre 2007). Ed. Antonio Cerviño Eiroa, Alejandro Guerra Díaz y Carmen Pérez Acosta. [EN] The ontogeny of the digestive tract during the larval development of cophthalmus rhombus was studied using histological and histochemical techniques. At exogenous feeding (4 days after hatching –dah-), both the mouth and the anus are opened and the digestive tract was differentiated in bucopharyngeal cavity, oesophagus, incipient stomach, foregut, midgut and hindgut. The gastric glands differentiated around 23 dah. Digestive mucous cells were rich in neutral and/or acidic mucosubstances. Intestinal lipid absorption was evident from 7-8 dah and protein supranuclear vesicles (protein micropynocitosis) of the hindgut were detected at 4 dah. The exocrine pancreas shows, around 2-3 dah, acidophilic zymogen granules (enzymatic precursors) and at the same time the hepatic glycogen granules and endocrine pancreas were identified from 2 dah. [ES] Se estudia la ontogenia del tracto digestivo durante el desarrollo larvario de parracho, Scophthalmus rhombus desde la eclosión hasta el día 38, utilizando técnicas histológicas e histoquímicas. Durante la fase de alimentación exógena (4 dde –días después de la eclosión-), tanto la boca como el ano están abiertos y el tracto digestivo está diferenciado en cavidad bucofaríngea, esófago, estómago incipiente e intestinos anterior, medio y posterior. Las glándulas gástricas se diferencian alrededor de los 23 dde e incrementan en número y desarrollo progresivamente. Las células mucosas digestivas son ricas en mucosustancias neutras y/o ácidas. Además, la absorción intestinal lipídica es evidente a los 7-8 dde y las vacuolas supranucleares (micropinocitosis proteica) del intestino posterior se detectaron a los 4 dde. El páncreas exocrino presenta gránulos de zimógeno acidófilos (precursores enzimáticos) a los 2-3 dde y al mismo tiempo se identifican los gránulos de glucógeno hepáticos y el páncreas endocrino. Este trabajo ha sido financiado por los proyectos DIVERAQUA y REDAQUAb del IFAPA (Interreg IIIA: SP5.E36 y SP5.E27/02) y parcialmente por el CSIC (AGL2005-02478 y AGL2006-13777-CO3-O2/ACU).

4 páginas, 2 figuras. XI Congreso Nacional de Acuicultura (Vigo, 24-28 septiembre 2007). Ed. Antonio Cerviño Eiroa, Alejandro Guerra Díaz y Carmen Pérez Acosta., [EN] The ontogeny of the digestive tract during the larval development of cophthalmus rhombus was studied using histological and histochemical techniques. At exogenous feeding (4 days after hatching –dah-), both the mouth and the anus are opened and the digestive tract was differentiated in bucopharyngeal cavity, oesophagus, incipient stomach, foregut, midgut and hindgut. The gastric glands differentiated around 23 dah. Digestive mucous cells were rich in neutral and/or acidic mucosubstances. Intestinal lipid absorption was evident from 7-8 dah and protein supranuclear vesicles (protein micropynocitosis) of the hindgut were detected at 4 dah. The exocrine pancreas shows, around 2-3 dah, acidophilic zymogen granules (enzymatic precursors) and at the same time the hepatic glycogen granules and endocrine pancreas were identified from 2 dah., [ES] Se estudia la ontogenia del tracto digestivo durante el desarrollo larvario de parracho, Scophthalmus rhombus desde la eclosión hasta el día 38, utilizando técnicas histológicas e histoquímicas. Durante la fase de alimentación exógena (4 dde –días después de la eclosión-), tanto la boca como el ano están abiertos y el tracto digestivo está diferenciado en cavidad bucofaríngea, esófago, estómago incipiente e intestinos anterior, medio y posterior. Las glándulas gástricas se diferencian alrededor de los 23 dde e incrementan en número y desarrollo progresivamente. Las células mucosas digestivas son ricas en mucosustancias neutras y/o ácidas. Además, la absorción intestinal lipídica es evidente a los 7-8 dde y las vacuolas supranucleares (micropinocitosis proteica) del intestino posterior se detectaron a los 4 dde. El páncreas exocrino presenta gránulos de zimógeno acidófilos (precursores enzimáticos) a los 2-3 dde y al mismo tiempo se identifican los gránulos de glucógeno hepáticos y el páncreas endocrino., Este trabajo ha sido financiado por los proyectos DIVERAQUA y REDAQUAb del IFAPA (Interreg IIIA: SP5.E36 y SP5.E27/02) y parcialmente por el CSIC (AGL2005-02478 y AGL2006-13777-CO3-O2/ACU).