The complete mitochondrial genome of Octopus vulgaris.

BACKGROUND: The Octopus vulgaris species complex consists of numerous morphologically similar but genetically distinct species. The current publicly available mitogenome of this species has been generated from a specimen collected from Tsukiji Fish Market, Tokyo, Japan. Octopus from the northwestern Pacific Ocean are now considered to be a separate species, Octopus sinensis. For this reason, we hypothesised that the current record of O. vulgaris was sequenced from a specimen of O. sinensis. Here, we sequenced the first complete mitogenome of a specimen of Octopus vulgaris sensu stricto that was collected from the species' confirmed distribution areas in northeastern Atlantic. METHODS AND RESULTS: The complete mitogenome was assembled de novo and annotated using 250 bp paired-end sequences. A single circular contig 15,655 bp in length with a mean read coverage of 1089 reads was reconstructed. The annotation pipeline identified 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNA) and two ribosomal RNAs. A maximum likelihood phylogenetic tree recovered the assembled mitogenome as the sister taxon of a monophyletic group comprising O. sinensis and the previously published mitogenome of "O. vulgaris" from Japan. This confirms that the latter was a Japanese specimen of O. sinensis. CONCLUSION: The mitogenome sequenced here is the first to be published for Octopus vulgaris sensu stricto. It represents an important first step in genetics-informed research on the evolution, conservation, and management of this commercially important species.

Trophic ecology of Octopus vulgaris paralarvae along the Iberian Canary current eastern boundary upwelling system.

Our knowledge of the diet of wild octopus paralarvae, Octopus vulgaris, is restricted to the first 2 weeks of its planktonic phase when they are selective hunters found near the coastline. These small paralarvae, bearing only three suckers per arm, are transported by oceanic currents from the coast towards offshore waters, where they complete the planktonic phase over 2 months. Here, we have investigated the trophic ecology of O. vulgaris paralarvae in two contrasting upwelling sub-regions of the Iberian Canary current (ICC) eastern boundary upwelling system and have evaluated dietary change as paralarvae develop (inferred by counting the number of suckers per arm, ranging from three to 15) along the coastal-oceanic gradient during their planktonic phase. Using high-throughput amplicon sequencing, we have characterised the diet of 100 paralarvae collected along the Northwest Iberian Peninsula (n = 65, three to five suckers per arm) and off the west coast of Morocco (n = 35, three to 15 suckers per arm), identifying up to 87 different prey species. The diet of paralarvae varied along the ICC, with crabs (53.4%), siphonophores (12.2%), copepods (12.3%), cnidarians (8.4%) and pteropods (3.7%) accounting for 90% of the variability detected off NW Iberian Peninsula, whereas off W Morocco, crabs (46.2%), copepods (23.1%), cnidarians (12.9%), krill (9.3%) and fishes (4.2%) explained 95.6% of the variability observed using frequency of observance (FOO%) data. Ontogenetic changes in the diet based on groups of paralarvae with similar numbers per arm were evidenced by the decreasing contribution of coastal meroplankton and an increase in oceanic holoplankton, including siphonophores, copepods, pteropods and krill. Trophic niche breadth values ranged from 0.06 to 0.67, with averaged values ranging from 0.23 to 0.33 (generalist = 1 and specialist = 0), suggesting that O. vulgaris paralarvae are selective predators through their ontogenetic transition between coastal and oceanic environments.

Small copepods could channel missing carbon through metazoan predation.

Global ecosystem models are essential tools for predicting climate change impacts on marine systems. Modeled biogenic carbon fluxes in the ocean often match measured data poorly and part of this could be because small copepods (<2 mm) are modeled as unicellular feeders grazing on phytoplankton and microzooplankton. The most abundant copepods from a seasonal upwelling region of the Eastern North Atlantic were sorted, and a molecular method was applied to copepod gut contents to evaluate the extent of metazoan predation under two oceanographic conditions, a trophic pathway not accounted for in global models. Scaling up the results obtained herein, based on published field and laboratory estimates, suggests that small copepods could ingest 1.79-27.20 gigatons C/year globally. This ignored metazoan-copepod link could increase current estimates of biogeochemical fluxes (remineralization, respiration, and the biological pump) and export to higher trophic levels by 15.6%-24.4%. It could also account for global discrepancies between measured daily ingestion and copepod metabolic demand/growth. The inclusion of metazoan predation into global models could provide a more realistic role of the copepods in the ocean and if these preliminary data hold true at larger sample sizes and scales, the implications would be substantial at the global scale.

New Data on the Systematics of Comb-fin Squids Chtenopteryx spp. (Cephalopoda: Chtenopterygidae) from the Canary Islands.

Alejandro Escánez, Álvaro Roura, Rodrigo Riera, Ángel Francisco González, and Ángel Guerra (2018) The systematics of the comb-fin squid species is problematic and poorly resolved. In total, 53 specimens of comb-fin squids (Chtenopteryx spp.) were caught at depths ranging from 30 to 800 m off the Canary Islands (NE Atlantic Ocean). Mantle lengths of the individuals ranged from 18 to 43 mm and the sample included immature, mature male and mature female specimens. Two species of comb-fin squids, Chtenopteryx canariensis and C. sicula, were identified by combining traditional morphological characters with a molecular analysis of a fragment of the cytochrome c oxidase subunit I (COI) gene. Intra- and interspecific genetic distances and maximum likelihood tree analyses based on COI sequences available from GenBank suggest the existence of at least four species, two from the Pacific and two from the Atlantic Ocean. Our data expand the current geographic range of C. canariensis from the NE to NW Atlantic. In the GenBank database, several sequences of comb-fin squid in different species-specific clades have been attributed only to C. sicula, indicating the possible existence of cryptic species and the need to re-analyse these data.

Prey Capture, Ingestion, and Digestion Dynamics of Octopus vulgaris Paralarvae Fed Live Zooplankton.

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 mantle contractions. Current results provide novel insights into both, O. vulgaris paralarvae-live prey capture strategies and the physiological mechanisms following ingestion, providing key information required to develop an effective rearing protocol for O. vulgaris paralarvae.

Feeding Relationship between Octopus vulgaris (Cuvier, 1797) Early Life-Cycle Stages and Their Prey in the Western Iberian Upwelling System: Correlation of Reciprocal Lipid and Fatty Acid Contents.

Under the influence of the Western Iberian upwelling system, the Iberian Atlantic coast holds important hatcheries and recruitment areas for Octopus vulgaris. Recently identified as an octopus hatchery, the Ría de Vigo harbors an important mesozooplankton community that supports O. vulgaris paralarvae during the first days of their planktonic stage. This study represents a preliminary approach to determine the nutritional link between wild O. vulgaris hatchlings, paralarvae and their zooplankton prey in the Ría de Vigo, by analyzing their lipid class content and fatty acid profiles. The results show that octopus hatchlings are richer in structural lipids as phospholipids and cholesterol, while the zooplankton is richer in reserve lipids like triacylglycerol and waxes. Zooplankton samples are also particularly rich in C18:1n9 and 22:6n3 (DHA), that seem to be successfully incorporated by O. vulgaris paralarvae thus resulting in a distinct fatty acid profile to that of the hatchlings. On the other hand, content in C20:4n6 (ARA) is maintained high through development, even though the zooplankton is apparently poorer in this essential fatty acid, confirming its importance for the development of O. vulgaris paralarvae. The content in monounsaturated fatty acids, particularly C18:1n7, and the DHA: EPA ratio are suggested as trophic markers of the diet of O. vulgaris paralarvae.

You Are What You Eat: A Genomic Analysis of the Gut Microbiome of Captive and Wild Octopus vulgaris Paralarvae and Their Zooplankton Prey.

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.

Diet Composition and Variability of Wild Octopus vulgaris and Alloteuthis media (Cephalopoda) Paralarvae: a Metagenomic Approach.

The high mortality of cephalopod early stages is the main bottleneck to grow them from paralarvae to adults in culture conditions, probably because the inadequacy of the diet that results in malnutrition. Since visual analysis of digestive tract contents of paralarvae provides little evidence of diet composition, the use of molecular tools, particularly next generation sequencing (NGS) platforms, offers an alternative to understand prey preferences and nutrient requirements of wild paralarvae. In this work, we aimed to determine the diet of paralarvae of the loliginid squid Alloteuthis media and to enhance the knowledge of the diet of recently hatched Octopus vulgaris paralarvae collected in different areas and seasons in an upwelling area (NW Spain). DNA from the dissected digestive glands of 32 A. media and 64 O. vulgaris paralarvae was amplified with universal primers for the mitochondrial gene COI, and specific primers targeting the mitochondrial gene 16S gene of arthropods and the mitochondrial gene 16S of Chordata. Following high-throughput DNA sequencing with the MiSeq run (Illumina), up to 4,124,464 reads were obtained and 234,090 reads of prey were successfully identified in 96.87 and 81.25% of octopus and squid paralarvae, respectively. Overall, we identified 122 Molecular Taxonomic Units (MOTUs) belonging to several taxa of decapods, copepods, euphausiids, amphipods, echinoderms, molluscs, and hydroids. Redundancy analysis (RDA) showed seasonal and spatial variability in the diet of O. vulgaris and spatial variability in A. media diet. General Additive Models (GAM) of the most frequently detected prey families of O. vulgaris revealed seasonal variability of the presence of copepods (family Paracalanidae) and ophiuroids (family Euryalidae), spatial variability in presence of crabs (family Pilumnidae) and preference in small individual octopus paralarvae for cladocerans (family Sididae) and ophiuroids. No statistically significant variation in the occurrences of the most frequently identified families was revealed in A. media. Overall, these results provide new clues about dietary preferences of wild cephalopod paralarvae, thus opening up new scenarios for research on trophic ecology and digestive physiology under controlled conditions.

Cannibalistic behavior of octopus (Octopus vulgaris) in the wild.

The first description of cannibalism in wild adult Octopus vulgaris is presented from 3 observations made in the Ría de Vigo (NW Spain), which were filmed by scuba divers. These records document common traits in cannibalistic behavior: (a) it was intercohort cannibalism; (b) attacks were made by both males and females; (c) in 2 of the records, the prey were transported to the den, which was covered with stones of different sizes; (d) the predator started to eat the tip of the arms of its prey; (e) predation on conspecifics occurred even if there were other abundant prey available (i.e., mussels); and (f) the prey/predator weight ratio in the 3 cases ranged from 20% to 25% body weight. The relationships between this behavior and sex, defense of territory, energy balance, food shortage, competition and predation, as well as how the attacker kills its victim are discussed.

Sperm ultrastructure in Bathypolypus bairdii and B. sponsalis (Cephalopoda: Octopoda).

A morphological comparison of the spermatozoa of the octopods Bathypolypus bairdii and B. sponsalis has been carried out by electron microscopy. Although the mature spermatozoon of B. bairdii is longer and thinner than that of B. sponsalis, its general ultrastructure is similar except for some minor differences. Their characteristic acrosomes, described here for the first time, consist of a periodically banded cone surrounded by a double helix whose arrangement has been defined by a numeric expression. The plasma membrane of sponsalis that surrounds the acrosome has many projections filled with abundant granular cytoplasm, which appears as a star-shaped acrosome in cross-section. The mitochondrial sheath contains 9-11 mitochondria in bairdii but only 9 in sponsalis. A comparison with other Octopodidae shows that the nucleus of both species is the largest ever seen. Our results support the capability of sperm morphology to discriminate between species and could even discern at higher taxonomic levels.