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Front Physiol
2017 May 24;8:321. doi: 10.3389/fphys.2017.00321.
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Diet Composition and Variability of Wild Octopus vulgaris and Alloteuthis media (Cephalopoda) Paralarvae: a Metagenomic Approach.
Olmos-Pérez L
,
Roura Á
,
Pierce GJ
,
Boyer S
,
González ÁF
.
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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.
Figure 1. (A) Map of the study area showing the four transects performed in 2012 and 2014. (B) Depth layers sampled for each different transects.
Figure 2. Proportion of Molecular Operational Taxonomic Units (MOTU) prey reads (PR) detected with the primer COI (A) and the primer 16Sa (B) in O. vulgaris and A. media. MOTUs were clustered in orders (A) or families (B).
Figure 3. Frequency of the occurrence of the families (FF) detected in O. vulgaris with primers COI (A) and primers 16Sa (B) with primer COI. Colors represent different orders. The corresponding phylum is indicated on the left. Vertical dashed line indicates the families detected in more than 10% of paralarvae.
Figure 4. Frequency of the occurrence of families (FF) detected in A. media with primers COI (A) and with primer 16Sa (B). Colors represent different orders. The corresponding phylum is indicated on the left. Vertical dashed line indicates the families detected in more than 10% of paralarvae.
Figure 5. (A) RDA triplot for the O. vulgaris families identified with COI gene. The correlation matrix was used. The first axis explains 29.57% and the second axis explains 21.96% of the total sum of all canonical eigenvalues (0.192). All the explanatory variables were used. (B) RDA triplot for the A. media families identified with COI gene. The correlation matrix was used. The first axis explains 27.26% and the second axis explains 21.47% of the total sum of all canonical eigenvalues (0.39). All the explanatory variables were used.
Figure 6. Families detected in O. vulgaris paralarvae (A) and A. media paralarvae (B) in different transects (2, 3 4, 5) and different seasons (summer, autumn). The vertical axis represents the number of paralarvae that present a given family.
Figure 7. Smooth curves for partial effects obtained by the Generalized Additive Modeling (GAM) of the occurrence of the families Pilumnidae (A), Euryalidae (B), Sididae (C,D), and Paracalanidae (E) as prey of O. vulgaris paralarvae. Explanatory variables used were dorsal mantle length (DML) and depth (Z1, Z2, Z3, Z4). Dotted lines are 95% confidence bands. The vertical axis represents the effect on the response variable.
Figure 8. Summary from 10 randomized sample sets of species identified in O. vulgaris with primers COI (A), primers 16Sa (B), and both pair of primers (C). Average values are presented as continues lines and Confidence Intervals (C.I.) as dashed lines.
Figure 9. Summary from 10 randomized sample sets of species identified in A. media with primers COI (A), primers 16Sa (B), and both pair of primers (C). Average values are presented as continues lines and Confidence Intervals (C.I.) as dashed lines.
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