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Research Project
Towards cephalopod aquaculture: cuttlefish as the spearhead species
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Control of zootechnology leads to improved Cuttlefish (Sepia officinalis, L.) reproduction performance up to pre-industrial levels
Publication . Capaz, Juan Carlos; Hernández-Brooke, Daniel; Balvet, Safia; Couto, Ana T.; Alves, Alexandra; Baptista Gonçalves, Rui; Frias, Paulo A.; Andrade, José Pedro; Sykes, António
Cephalopods are gaining momentum as an alternate group for aquaculture species diversification, not only because they are a good food source (highly appreciated in some worldwide markets) but they also have the potential to quickly reach a market size. However, there are some bottlenecks impeding the transition of culture technology from the laboratory to industry. One is related to control over reproduction in captivity. The objective of the present experiment was to verify the effects of tanks with different bottom areas/volumes on the reproduction performance of S. officinalis breeding stocks, when sex ratios were controlled a priori; and the food cost associated with such performance when individuals are fed a natural frozen diet. One hundred and ninety two juvenile cuttlefish were used to compare three different round-shaped tanks: one type with 3000L volume and two types with 9000L volume (with differences in bottom areas and water column). Individuals had their sex and maturity stage determined to establish a sexual ratio of 2 female:1 male per tank and assure that cuttlefish were still immature. Biological data was collected during both growth and reproduction stages and until the death of all females in each tank. The experiment lasted nearly 300 days. Temperature differences between tank types were registered during both stages. The optimizing of rearing conditions has allowed for higher growth and a higher amount of cuttlefish available for breeding purposes. A total of 123,751 eggs (in 85 batches) was obtained during this experiment, which is a number that may meet a small scale cuttlefish commercial hatchery facility requirements. The present conditions contributed to a better and predictable reproduction performance in specific 9000L tanks, with values reaching pre-industrial numbers (approximate to 24,000 eggs/tank). Moreover, both the amount of eggs per batch and the overall quality of eggs has increased. Three of these 9000L tanks have an overall consumption of approximate to 38.64 Kg tank(-1), which translates in an investment in feed of approximate to 193 (sic) tank(-1), 8.40 (sic) per cuttlefish and an overall daily tank expense of 1.76 (sic) d(-1).
In vivo metabolism of unsaturated fatty acids in Sepia officinalis hatchlings
Publication . Reis, Diana; Rodriguez, C.; Acosta, N. G.; Almansa, E.; Tocher, D. R.; Andrade, José Pedro; Sykes, António V.
The transition of Sepia officinalis culture to industrial large scale has been hampered due to bottlenecks related to the limited knowledge on nutritional physiology of the species. Determination of the endogenous ability of S. officinalis hatchlings to metabolise unsaturated fatty acids (FA) may provide new insight on the capability of hatchlings to biosynthesise different FA, as well as lipid classes containing essential fatty acids (EFA). In the present study, cuttlefish hatchlings were incubated with [1-C-14] FA including C18 FA (18:1n-9, 18:2n-6, 18:3n-3) and long-chain polyunsaturated fatty acids (LC-PUFA) (20: 4n-6 (ARA), 20:5n-3 (EPA) or 22:6n-3 (DHA)), which were added individually as potassium salts bound to bovine serumalbumin. As a result, it was possible to investigate the in vivo FAmetabolismof S. officinalis hatchlings by following the incorporation of specific [1-C-14] FA, which points to the suitability of this methodology to study lipid metabolism of newly hatched cephalopods. The majority of radioactivity incorporated was recovered esterified into polar lipids (PL). A pattern was detected, where [1-C-14] DHA, [1-C-14] C18 FA and their metabolic products were preferentially esterified into phosphatidylcholine, whereas [1-C-14] ARA and [1-C-14] EPA were mainly esterified into phosphatidylethanolamine. [1-C-14] C18 FA were the most transformed FA with several metabolites produced by elongation and possible desaturation being obtained. As a contrary the radioactivity incorporated into hatchling total lipid (TL) from supplemented [1-C-14] LC-PUFA only one elongation product was recovered from the three substrates, except for [1-C-14] ARA, where an unidentified product was also detected. The present in vivo results indicated that S. officinalis hatchlings may have capability for the first steps in the biosynthesis of ARA and EPA from 18: 2n-6 and 18: 3n-3, respectively, including the existence of a desaturase potentially involved. Nonetheless, considering the low desaturation rates detected, this process may not be sufficient to cover EFA demands during development of this species. Therefore, dietary ARA and EPA, as well as DHA, should be supplied during the hatchling stage of Sepia. While designing an inert diet, which ensures normal growth and development of this species during the hatchling stage, the C18 FA and LC-PUFA levels and ratios should be considered, since the esterification pattern detected in the present study suggested competition between these FA for esterification into specific lipid classes. Moreover, considering the observed esterification pattern of LC-PUFA into different lipid classes, it is likely that the DHA/EPA/ARA ratio, rather than DHA/EPA or EPA/ARA ratios, would be of great importance for S. officinalis hatchling development. (C) 2015 Elsevier B.V. All rights reserved.
Behavioural aspects of the spotty bobtail squid Euprymna parva (Cephalopoda: Sepiolidae)
Publication . Drerup, Christian; Sykes, António; Cooke, Gavan M.
Bobtail squids (Sepiolidae, Cephalopoda) have recently been growing in popularity in scientific studies due to their symbiotic relationship with light producing bacteria and their corresponding light emitting organs. However, the overall knowledge on the behaviour of sepiolids is based on observations on just a few of the roughly 70 extant species and must still be considered as sparsely. Understanding their behavioural ecology is not only beneficial to further grasp the complex behavioural patterns of cephalopods, it is also vital for establishing a good welfare practice when holding sepiolids in captivity. Hence, the present study characterised several behavioural aspects of the spotty bobtail squid Euprymna parva. Although the burying, hunting and mating behaviour as well as most escape responses of this less investigated sepiolid species greatly resembled those of other observed bobtail squids, differences to sepiolids from other genera or even from the same genus could be identified in the present study. Additionally, the first observation of an up to now undescribed inking behaviour of sepiolids is reported. E. parva was observed to eject a stretch of ink ('ink rope'), approximately 4-5 times the length of the animal, and hold on to it motionless, potentially as a masquerade to resemble a floating seagrass leave. The present study further provides detailed information on daily time and activity budgets as well as the tentacular strike speed during hunting, two up to now barely investigated behavioural aspects of the sepiolid ecology.
Preliminary results on the daily and seasonal rhythms of cuttlefish Sepia officinalis (Linnaeus, 1758) locomotor activity in captivity
Publication . Oliveira, Catarina; Grano-Maldonado, Mayra; Gonçalves, Rui A.; Frias, Paulo; Sykes, António V.
Cephalopods are considered as an alternate group for aquaculture species diversification [1,2], not only because they are a good food source (highly appreciated in some worldwide markets, e.g., Portugal, Spain, Italy and Asia) and have the potential to quickly reach a market size, but also because they are considered animal models in several fields of research [3–5], and it is expected that such models are progressively obtained from bioteriums instead of being captured from nature.
Despite the recent advances in culture protocols of several cephalopod species [2], there is still little information available about the biological rhythms of cephalopods in captivity. In fact, to the best of our knowledge, there are only 4 published papers concerning this theme, and these are focused on octopus species [6–9]. This issue assumes increasing importance with the enforcement of EU welfare legislation (Directive 2010/63/EU), and this information is critical for the future update of Annex III (housing conditions) for cephalopods of that Directive, and for a future update of the Guidelines for the Care and Welfare of Cephalopods in Research [10]. If it is considered that the European cuttlefish Sepia officinalis (Linnaeus, 1758) may have, at the least, two different sleep‐like states [11], the importance of this study becomes even more evident.
Felasa working group report: capture and transport of live cephalopods – recommendations for scientific purposes
Publication . Sykes, António; Galligioni, Viola; Estefanell, Juan; Hetherington, Stuart; Brocca, Marco; Correia, Joao; Ferreira, André; Pieroni, Eleonora M.; Fiorito, Graziano
On 1 January 2013, research using cephalopod molluscs, from hatchlings to adults, became regulated within
Directive 2010/63/EU. There are significant difficulties in captive breeding in the great majority of currently
utilised species. Thus, scientific research relies upon the use of wild-caught animals. Furthermore, live
cephalopods are shared and transported between different stakeholders and laboratories across Europe
and other continents. Despite existing European and national legislation, codes, guidelines and reports
from independent organisations, a set of recommendations specifically addressing the requirements for
the capture and transport of animals belonging to this taxon are missing. In addition, although training
and development of competence for all people involved in the supply chain are essential and aim to
ensure that animals do not suffer from pain, distress or lasting harm, the requirements for those capturing
and transporting wild cephalopods have not been considered. This Working Group reviewed the current
literature to recognise scientific evidence and the best practice, and compiled a set of recommendations
to provide guidance on the ‘techniques’ to be used for the capture and transport of live cephalopods for their
use in scientific procedures. In addition, we propose to (a) develop standardised approaches able to assess
recommended methods and objectively quantify the impact of these processes on animals’ health, welfare
and stress response, and (b) design a training programme for people attaining the necessary competence for
capture and transportation of live cephalopods, as required by Directive 2010/63/EU.
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Funding agency
Fundação para a Ciência e a Tecnologia
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Investigador FCT
Funding Award Number
IF/00576/2014/CP1217/CT0002