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The European sardine (Sardina pilchardus, Walbaum 1792) is indisputably a commercially important species. Previous studies using uneven sampling or a limited number of makers have presented sometimes conflicting evidence of the genetic structure of S. pilchardus populations. Here, we show that whole genome data from 108 individuals from 16 sampling areas across 5000 km of the species’ distribution range (from the Eastern Mediterranean to the archipelago of Azores) support at least three genetic clusters. One includes individuals from Azores and Madeira, with evidence of substructure separating these two archipelagos in the Atlantic. Another cluster broadly corresponds to the center of the distribution, including the sampling sites around Iberia, separated by the Almeria–Oran front from the third cluster that includes all of the Mediterranean samples, except those from the Alboran Sea. Individuals from the Canary Islands appear to belong to the Mediterranean cluster. This suggests at least two important geographical barriers to gene flow, even though these do not seem complete, with many individuals from around Iberia and the Mediterranean showing some patterns compatible with admixture with other genetic clusters. Genomic regions corresponding to the top outliers of genetic differentiation are located in areas of low recombination indicative that genetic architecture also has a role in shaping population structure. These regions include genes related to otolith formation, a calcium carbonate structure in the inner ear previously used to distinguish S. pilchardus populations. Our results provide a baseline for further characterization of physical and genetic barriers that divide European sardine populations, and information for transnational stock management of this highly exploited species towards sustainable fisheries.

Egg specific gravity is of relevance for fish recruitment since the ability to float influences egg and larvae development, dispersal and connectivity between fishing grounds. Using zootechnics, histological approaches, optical and electronic transmission microscopy, this study describes the morphogenetic mechanism of adhesion of the oil-drop covering layer (OCL) to the oil droplet (OD) in embryos of Merluccius merluccius under physical conditions reflecting the marine environment. The herein described primordial (p)OCL is a substructure of the inner yolk syncytial layer which contains egg organella aimed to mobilize lipidic reserves from the oil drop (OD) towards the embryo blood. It is shown that the timely OD-OCL assembly is a critical morphogenetic process for embryo and larvae survival. Such assembly depends on egg buoyance because of its influence on the embryo capacity to rotate within the perivitelline space. Therefore, oil droplet adhesion (ODA) eggs are capable to complete their development while oil droplet non-adhesion eggs (ODNA) dye soon after hatching. We show that gravity-dependent egg buoyance categories exhibit different ODA/ODNA ratios (0–77%) and that relationship diminishes under incubation systems such as sprayers, that do not assure a dynamic seawater surface mixing to avoid egg desiccation. As an adaptive trait, egg gravity strongly depends on oceanic properties such as current dynamics, turbulence, oxygen, rainfall, and salinity, whose rapid changes would likely challenge the sustainability of fisheries recruitment.

The European sardine (Sardina pilchardus, Walbaum 1792) is indisputably a commercially important species. Previous studies using uneven sampling or a limited number of makers have presented sometimes conflicting evidence of the genetic structure of S. pilchardus populations. Here, we show that whole genome data from 108 individuals from 16 sampling areas across 5000 km of the species' distribution range (from the Eastern Mediterranean to the archipelago of Azores) support at least three genetic clusters. One includes individuals from Azores and Madeira, with evidence of substructure separating these two archipelagos in the Atlantic. Another cluster broadly corresponds to the center of the distribution, including the sampling sites around Iberia, separated by the Almeria-Oran front from the third cluster that includes all of the Mediterranean samples, except those from the Alboran Sea. Individuals from the Canary Islands appear to belong to the Mediterranean cluster. This suggests at least two important geographical barriers to gene flow, even though these do not seem complete, with many individuals from around Iberia and the Mediterranean showing some patterns compatible with admixture with other genetic clusters. Genomic regions corresponding to the top outliers of genetic differentiation are located in areas of low recombination indicative that genetic architecture also has a role in shaping population structure. These regions include genes related to otolith formation, a calcium carbonate structure in the inner ear previously used to distinguish S. pilchardus populations. Our results provide a baseline for further characterization of physical and genetic barriers that divide European sardine populations, and information for transnational stock management of this highly exploited species towards sustainable fisheries.

Common octopus, Octopus vulgaris, is an economically important cephalopod species. However, its rearing under captivity is currently challenged by massive mortalities previous to their juvenile stage due to nutritional and environmental factors. Dissecting the genetic basis and regulatory mechanism behind this mortality requires genomic background knowledge. A transcriptomic sequencing of 10 dph octopus paralarvae from different experimental conditions was constructed via RNA-seq. A total of 613,767,530 raw reads were filtered and de novo assembled into 363,527 contigs of which 82,513 were annotated in UniProt carrying also their GO and KEGG information. Differential gene expression analysis was carried out on paralarvae reared under different diet regimes and temperatures, also including wild paralarvae. Genes related to lipid metabolism exhibited higher transcriptional levels in individuals whose diet includes crustacean zoeas, which had an impact over their development and immune response capability. High temperature induces acclimation processes at the time that increase metabolic demands and oxidative stress. Wild individuals show an expression profile unexpectedly similar to Artemia fed individuals. Proteomic results support the hypothesis revealed by transcriptional analysis. The comparative study of the O. vulgaris transcriptomic profiles allowed the identification of genes that deserve to be further studied as candidates for biomarkers of development and health. The results obtained here on the transcriptional variations of genes caused by diet and temperature will provide new perspectives in understanding the molecular mechanisms behind nutritional and temperature requirements of common octopus that will open new opportunities to deepen in paralarvae rearing requirements

Common octopus, Octopus vulgaris, is an economically important cephalopod species. However, its rearing under captivity is currently challenged by massive mortalities previous to their juvenile stage due to nutritional and environmental factors. Dissecting the genetic basis and regulatory mechanism behind this mortality requires genomic background knowledge. A transcriptomic sequencing of 10 dph octopus paralarvae from different experimental conditions was constructed via RNA-seq. A total of 613,767,530 raw reads were filtered and de novo assembled into 363,527 contigs of which 82,513 were annotated in UniProt carrying also their GO and KEGG information. Differential gene expression analysis was carried out on paralarvae reared under different diet regimes and temperatures, also including wild paralarvae. Genes related to lipid metabolism exhibited higher transcriptional levels in individuals whose diet includes crustacean zoeas, which had an impact over their development and immune response capability. High temperature induces acclimation processes at the time that increase metabolic demands and oxidative stress. Wild individuals show an expression profile unexpectedly similar to Artemia fed individuals. Proteomic results support the hypothesis revealed by transcriptional analysis. The comparative study of the O. vulgaris transcriptomic profiles allowed the identification of genes that deserve to be further studied as candidates for biomarkers of development and health. The results obtained here on the transcriptional variations of genes caused by diet and temperature will provide new perspectives in understanding the molecular mechanisms behind nutritional and temperature requirements of common octopus that will open new opportunities to deepen in paralarvae rearing requirements.

The common octopus Octopus vulgaris is a highly valuable species worldwide, but to understand its population dynamics and requirements under culture conditions, it is crucial to improve our knowledge about its planktonic stages. Previous studies validating daily beak growth increments in these stages allowed age estimation and comparison of wild and cultured paralarvae. We aimed to improve age estimations in captivity, addressing the effect of temperature and diet, to obtain an accurate estimation of age in wild specimens collected from the coast to the open ocean off NW Spain and Morocco. We analysed the beak growth increments of reared paralarvae at 14 and 21°C with 2 different crustacean prey taxa (Artemia and spider crab Maja brachydactyla zoeae) over 30 d. Daily increment deposition at 21°C was confirmed, whereas <1 increment d-1 was recorded at 14°C. The width of the reading area grew accordingly with age; therefore, this beak region may be suitable for age estimation. A general linear model (GLM) analysis showed that temperature and the interaction of age × temperature significantly influenced increment deposition, whereas diet did not. The number of growth rings recorded in wild paralarvae beaks ranged from 0-8 on the coast, 7-11 on the continental shelf and 2-28 in the open ocean. Corrected age estimates of wild paralarvae were obtained with the GLM using the mean temperatures recorded in the wild, supporting the hypothesis that O. vulgaris leave the coastal area and develop in the open ocean transported by upwelling filaments.

The common octopus Octopus vulgaris is a highly valuable species worldwide, but to understand its population dynamics and requirements under culture conditions, it is crucial to improve our knowledge about its planktonic stages. Previous studies validating daily beak growth increments in these stages allowed age estimation and comparison of wild and cultured paralarvae. We aimed to improve age estimations in captivity, addressing the effect of temperature and diet, to obtain an accurate estimation of age in wild specimens collected from the coast to the open ocean off NW Spain and Morocco. We analysed the beak growth increments of reared paralarvae at 14 and 21°C with 2 different crustacean prey taxa (Artemia and spider crab Maja brachydactyla zoeae) over 30 d. Daily increment deposition at 21°C was confirmed, whereas <1 increment d−1 was recorded at 14°C. The width of the reading area grew accordingly with age; therefore, this beak region may be suitable for age estimation. A general linear model (GLM) analysis showed that temperature and the interaction of age × temperature significantly influenced increment deposition, whereas diet did not. The number of growth rings recorded in wild paralarvae beaks ranged from 0−8 on the coast, 7−11 on the continental shelf and 2−28 in the open ocean. Corrected age estimates of wild paralarvae were obtained with the GLM using the mean temperatures recorded in the wild, supporting the hypothesis that O. vulgaris leave the coastal area and develop in the open ocean transported by upwelling filaments.

This study describes the effect of seasonal average temperatures (14 and 18°C) in the Ría of Vigo, on the utilization of external yolk over the last five Naef stages of development (XV–XX) for Octopus vulgaris embryos. Also, the transference of the outer yolk to the inner yolk sac, and its use during embryonic development and early life by O. vulgaris paralarvae. Temperature had a marked effect on embryonic development, except during stages XV–XIX (until the second inversion) where development time was the same (14 days), regardless of temperature. There were no significant differences in outer yolk decrease between consecutive Naef stages at 14°C and 18°C. Contrary, significant differences at all Naef stages from XV to XIX (both, with or without outer yolk) were observed for inner yolk between temperatures. A higher accumulation of inner yolk in embryos at 14°C was observed, due to lower yolk consumption. Paralarvae incubated at both temperatures were maintained independently at starvation during 4 days. At 18°C, a reduced accumulation of inner yolk, especially during Naef stage XIX, was observed. In 24 h old paralarvae, there was already significant higher inner yolk content at 14°C than at 18°C. Unfed paralarvae at 18°C lost weight faster than those at 14°C, due to higher energetic requirements. Finally, from these results, we propose a paralarvae rearing protocol during the first days after hatching and during the last five Naef stage (XV–XX) at lower temperatures, since the energy requirements are lower during the initial maturation stage.

Octopus vulgaris is a species of great interest in research areas such as neurobiology, ethology, and ecology but also a candidate species for aquaculture as a food resource and for alleviating the fishing pressure on its wild populations. This study aimed to characterize the predatory behavior of O. vulgaris paralarvae and to quantify their digestive activity. Those processes were affordable using the video-recording analysis of 3 days post-hatching (dph), mantle-transparent paralarvae feeding on 18 types of live zooplanktonic prey. We show for the first time in a live cephalopod that octopus paralarvae attack, immobilize, drill, and ingest live cladocerans and copepods with 100% efficiency, which decreases dramatically to 60% on decapod prey (Pisidia longicornis). The majority (85%) of successful attacks targeted the prey cephalothorax while unsuccessful attacks either targeted the dorsal cephalothorax or involved prey defensive strategies (e.g., juvenile crab megalopae) or prey protected by thick carapaces (e.g., gammaridae amphipods). After immobilization, the beak, the buccal mass and the radula were involved in exoskeleton penetration and content ingestion. Ingestion time of prey content was rapid for copepods and cladocerans (73.13 ± 23.34 s) but much slower for decapod zoeae and euphausiids (152.49 ± 29.40 s). Total contact time with prey was always <5 min. Contrary to the conventional view of crop filling dynamics observed in adult O. vulgaris, food accumulated first in the stomach of paralarvae and the crop filled after the stomach volume plateaued. Peristaltic crop contractions (~18/min) moved food into the stomach (contractions ~30/min) from where it passed to the caecum. Pigmented food particles were seen to enter the digestive gland, 312 ± 32 s after the crop reached its maximum volume. Digestive tract contents passed into the terminal intestine by peristalsis (contraction frequency ~50/min) and defaecation was accompanied by an increased frequency of

The common octopus (Octopus vulgaris) is an attractive species for aquaculture, however, several challenges inhibit sustainable commercial production. Little is known about the early paralarval stages in the wild, including diet and intestinal microbiota, which likely play a significant role in development and vitality of this important life stage. High throughput sequencing was used to characterize the gastrointestinal microbiome of wild O. vulgaris paralarvae collected from two different upwelling regions off the coast of North West Spain (n = 41) and Morocco (n = 35). These were compared to that of paralarvae reared with Artemia for up to 25 days in captivity (n = 29). In addition, the gastrointestinal microbiome of zooplankton prey (crabs, copepod and krill) was also analyzed to determine if the microbial communities present in wild paralarvae are derived from their diet. Paralarvae reared in captivity with Artemia showed a depletion of bacterial diversity, particularly after day 5, when almost half the bacterial species present on day 0 were lost and two bacterial families (Mycoplasmataceae and Vibrionaceae) dominated the microbial community. In contrast, bacterial diversity increased in wild paralarvae as they developed in the oceanic realm of both upwelling systems, likely due to the exposure of new bacterial communities via ingestion of a wide diversity of prey. Remarkably, the bacterial diversity of recently hatched paralarvae in captivity was similar to that of wild paralarvae and zooplankton, thus suggesting a marked effect of the diet in both the microbial community species diversity and evenness. This study provides a comprehensive overview of the bacterial communities inhabiting the gastrointestinal tract of O. vulgaris paralarvae, and reveals new research lines to challenge the current bottlenecks preventing sustainable octopus aquaculture.

The fatty acid (FA) profile of wild Octopus vulgaris paralarvae of estimated age was individually analyzed for the very first time in order to establish a reference for comparison in rearing and nutritional studies. Age of each paralarvae was estimated by analysing daily increments on lateral hood surface of beaks. Wild paralarvae age ranged between 6 and 8 days and their FA composition resembled that from hatchlings produced under culture conditions. However, when compared with the FA composition of up to 20 days old cultured paralarvae described in the bibliography, some striking differences were found. Results showed higher levels of docosahexaenoic acid (22:6n − 3, DHA), lower contents of 18:1n − 9, 18:1n − 7 and 18:2n − 6 and negligible levels of 18:3n − 3 in wild paralarvae, when collated to reared one. These results seem to indicate that preys/diets supplied to cultured paralarvae fail to resemble paralarval natural composition and as a result do not fulfil their FA requirement. The individual applied technique developed in this study will allow to refine the study of wild paralarvae along its development, as well as to compare wild and cultured paralarvae of similar age.

This study describes the effect of seasonal average temperatures (14 and 18°C) in the Ría of Vigo, on the utilization of external yolk over the last five Naef stages of development (XV–XX) for Octopus vulgaris embryos. Also, the transference of the outer yolk to the inner yolk sac, and its use during embryonic development and early life by O. vulgaris paralarvae. Temperature had a marked effect on embryonic development, except during stages XV–XIX (until the second inversion) where development time was the same (14 days), regardless of temperature. There were no significant differences in outer yolk decrease between consecutive Naef stages at 14°C and 18°C. Contrary, significant differences at all Naef stages from XV to XIX (both, with or without outer yolk) were observed for inner yolk between temperatures. A higher accumulation of inner yolk in embryos at 14°C was observed, due to lower yolk consumption. Paralarvae incubated at both temperatures were maintained independently at starvation during 4 days. At 18°C, a reduced accumulation of inner yolk, especially during Naef stage XIX, was observed. In 24 h old paralarvae, there was already significant higher inner yolk content at 14°C than at 18°C. Unfed paralarvae at 18°C lost weight faster than those at 14°C, due to higher energetic requirements. Finally, from these results, we propose a paralarvae rearing protocol during the first days after hatching and during the last five Naef stage (XV–XX) at lower temperatures, since the energy requirements are lower during the initial maturation stage.

This study analyzes the effect of low salinity on survival and daily food ingestion of octopus subjected to three different water salinities (20, 25, and 30 psu) for 3 days. During the 3-day experiment, octopuses at 20 psu immediately stopped eating, and all died during the first day; at 25 and 30 psu, the octopuses gradually reduced their feed intake and stopped eating after 2 days, but all survived, and when normal salinity conditions were reestablished (34–35 psu), they ingested 30–40 (~27.5% of the initial food intake) and 70–90 g of food/day (~63% of the initial food intake) respectively over the 5-day recovery period. Water salinity of 30 psu could be considered as the recommended lower limit below which actions, such as transferring the commercial floating cages to another location or submerging them, should be taken in order to prevent the negative effects of low salinities on farmed octopuses.

Bajo condiciones de cultivo, la composición lipídica y, específicamente, el perfil de ácidos grasos de las paralarvas cultivadas, es significativamente diferente al de las paralarvas recién eclosionadas. Por tanto, la comparación entre paralarvas salvajes y cultivadas de edad equivalente, permitiría dilucidar si estos cambios detectados en el perfil de ácidos grasos están relacionados con una dieta inadecuada o son causados por el propio desarrollo de la paralarva. En el presente estudio, se analizó por primera vez de forma individual el perfil de ácidos grasos de paralarvas salvajes de Octopus vulgaris y se determinó su edad a través de la deposición diaria de los incrementos en los picos, con el objetivo de profundizar en los requerimientos lipídicos de las paralarvas a lo largo del desarrollo y de esta forma intentar optimizar la dieta de los ejemplares en cultivo.

Under culture conditions, lipid composition and specifically, the fatty acid profile of reared paralarvae is significantly different from hatchlings. Therefore, comparing wild paralarvae and reared paralarvae of similar age would allow us to elucidate if the changes in fatty acid profile are related with a non-optimal prey composition or are caused by the normal development. The aim of this study was analyze, for the very first time, the fatty acid profile of wild Octopus vulgaris paralarvae individually, determining for each paralarvae their age through deposition of daily increments on lateral hood surface of the beaks. The main goal is to clear up lipid requirements of paralarvae and optimise the diet of reared paralarvae.

In order to determine the number of individuals involved in breeding in a stock of hake (Merluccius merluccius), tissue samples were analyzed by genotyping of 5 microsatellites obtained from breeding individuals and larvae from three different spawns in 2011, 2012 and 2013. The results show a high polymorphism rates for the selected markers, as well as good scores of parental assignment. For each spawn, there is a large proportion of siblings and half-siblings individuals. These data suggest spawning events in which most of females and few males are involved, however more analysis are needed to determine in detail the reproductive dynamics of the stock., Con el fin de determinar el número de ejemplares que intervienen en la reproducción en un stock de merluza europea (Merluccius merluccius), se analizaron mediante genotipado de 5 microsatélites muestras obtenidas de los individuos reproductores, así como de las larvas de tres puestas diferentes en los años 2011, 2012 y 2013. Los resultados muestran un elevado polimorfismo para los marcadores seleccionados, así como unas buenas tasas de asignación de parentales. Para cada puesta, existe una gran proporción de individuos hermanos y medio-hermanos entre sí. Estos datos sugieren eventos de desove en los que participan la mayoría de las hembras y pocos machos, sin embargo se requieren más análisis para determinar en detalle la dinámica reproductiva del stock.