Advances in Environmental Science and Energy Planning

Effects of first feeding regimes on growth and survival of

( spp.)

L. MOLINA-DOMINGUEZ, A. LOZA, M. FERRAN, A. VILAR, and A. SEGADE IU-Ecoaqua University of Las Palmas de Gran Canaria Muelle de Taliarte s/n 35214 Telde (Las Palmas) SPAIN [email protected]

Abstract: Syngnathid populations are submitted to uncontrolled exploitation and inadequate management of their natural environment. Thus, culture of these species could contribute to satisfy their exhibition demand on public aquaria while supports the recovery of wild stocks. One of the main goals in modern aquaria is to complete the biological cycles of shown in their exhibitions, especially interesting in rare or threatened species showing high visitors’ interest. The present study was conducted to test the effect on growth and survival of different Artemia enrichments used as first feeding for the newborn pipefish (Syngnathus spp.). The four tested diets were: non-enriched Artemia, and three diets using enriched Artemia metanauplii using different products, two based on microalgae, Isochrysis galvana and Tetraselmis suecica, respectively, and the last one enriched on a commercial product, DHA protein SELCO (INVE, Belgium). Survival and growth parameters (size and weight) were studied along the trial, as well as data on water quality (temperature, dissolved oxygen, pH and salinity) were recorded.The results showed excellent survival (98% on average) in all the used diets, however differences on the growth rate has been observed. According to the results, juveniles fed on DHA protein-SELCO enriched Artemia metanauplii showed the highest size and weight at the end of the experiment.

Key-Words: First feeding, survival, growth, breeding, aquaria, pipefish.

1 Introduction Some investigations into the international trade in syngnathids revealed a large and growing trade in Pipefish, along with seahorses, are members of the seahorses, pipehorses, and for use in family . These species presents a large traditional medicines, aquarium , and distribution all over the Eastern Atlantic from curiosities [6],[7]. Norway to Senegal and from Namibia to Cape of Good Hope (South Africa); their presence has been The demand for various syngnathid has confirmed in Canary Islands [1]. It has been also been met traditionally by the collection of animals cited on Mediterranean coasts as Aegean and Black from the wild but the long term survival of natural seas [2]. Despite literature records, occurrence in the populations is likely to be jeopardised by an Indo-Pacific outside South African waters lacks increase in demand [8]. Nowadays, the conclusive evidence [3]. Knowledge on this family overexploitation of their stocks, it is now widely is generally scarce but has been increasing in recent acknowledged. Besides, species’ limited dispersal years due to their conservation status. Their status ability may make it more vulnerable to extinction has been the focus of increasing concern since the [9] especially if their are fragmented [10]. mid 1990s, both in their own right and as flagship In addition, several of the factors pointed to as species for broader marine conservation issues [4]. sources of endangerment for marine species – e.g., They are a charismatic group of fish that attracts a narrow distribution, vulnerability of its to high level of public support and sympathy [5]. destruction, and rarity – apply to the pipefish. For

ISBN: 978-1-61804-280-4 51 Advances in Environmental Science and Energy Planning

all these reasons, aquaculture must be considered as eicosapentaenoico (EPA; 20:5n-3) and an alternative to wild collection, especially for some docosahexaenoic acids (DHA; 22:6n-3) and these species of syngnathids [11], [12], [13]. must be supplied in the diet to ensure optimal growth and survival [19]. Because of that, the The presence of pipefish and other nutritional enrichment of live prey organisms before syngnathids in public aquaria is very common. The feeding is a common practice in aquaculture. popularity of these species is related on its look, attractiveness and its unusual reproductive Little is known about the nutritional behaviour, males have a brood pouch found beneath requirements of pipefish. In most species, the the anal fin, where females deposits their eggs and newborn fish are released from paternal pouch at a where are incubated until birth. The public aquaria later stage of development than occurs for other function is not only to provide entertainment and teleosts fish. It has not been established yet if relax to visitors and to increase their knowledge of newborn pipefish have the same requirements as environmental problems. Syngnathids have the other fish larvae. In the other hand, anesthetic potential to act as powerful icons for sustainable methods have generally been used in aquaculture to uses in public aquaria. These institutions can be reduce fish stress and improve handling [20], adopting environmental friendly and sustainable however its use for ornamental species is unusual. procedures, among them it can be stressed the There are scarce references relative of its use in development of technology for the rearing of Syngnathidae family in general and pipefish in ornamental species [14]. This statement is useful for particular. any ornamental specie but it can be stressed the case of the pipefish, focused species in this work because This work is focused in the , these species present natural spawning in aquaria. Syngnathus acus, however some data of Synghathus thyple were also presented. These species are very Pipefish are primary zooplanktivous; small common in public aquaria because the high level of crustaceans are the main food and in terms of public interest due its aspect and behavior. numerical and frequency of occurrence, as copepods, amphipods and crustacean larvae [15]. The main aim of this work is to test the These fish are not locked-in to the consumption of effects on survival and growth of newborn pipefish, specific prey but correlated by the seasonal of different enrichment (differing on n-3 HUFA abundance of different prey [16], [15]. The pipefish contents) of Artemia salina, the most commonly are a small species with a small mouth adapted for used live preys in aquaculture preying small animals using a strong inhalant current to capture them. With such small prey, 2 Material and Methods pipefish must consume large numbers of food organisms [8]. Because these reasons, the provision This study was carried out in the in the Aquarium on abundant and adequate live prey organisms is an Finisterrae in La Coruña (Spain). When pregnant essential tool for breeding these animals in captivity pipefish males were observed at exhibition aquaria, conditions. Artemia salina is one of the most clearly distinguished by the distended brood pouch, commonly used prey in aquaculture, as well as, in it was gently removed and transferred to a 30 l glass public aquaria, due to the existence of standardized aquarium and maintained at 20ºC on a diet base on cost-effective protocols for their mass production. Artemia until the offspring release. Batches released These live prey have been also used in syngnathid from a single male were divided into equal groups at breeding with different degrees of success, a stocking density of 5 pipefish L-1. For S. acus, depending upon size and species [17]. three batches were treated in the same way resulting three replicates for each diet. The experiment was Moreover, the success of fish larval rearing non-concurrent as batches were released at different is greatly influenced by first feeding regimes and the times [8]. In the case of S. thyple only one spawning nutritional quality of starter diets [18]. In this sense, episode occurred during the experimental period. the dietary lipids are recognized as one of the most The batch released was equally divided into three important nutritional factors that affect larval groups at a stocking density of 5 pipefish L-1 and growth and survival and in particular highly only T4 was tested. unsaturated fatty acids (HUFA). Numerous studies have shown that marine fish larvae have a dietary Feeding trials were conducted in 12x10 l requirement for arachidonic (ARA; 20:4n-6), polycarbonate containers supplied with flow-

ISBN: 978-1-61804-280-4 52 Advances in Environmental Science and Energy Planning

through ambient seawater, previously purified using essential oil (Guinama), containing 87% eugenol mechanical filtration and UV light (Philips, with seawater, from a diluted solution 0,5 % (v/v) in 25W/G25T8UV-C, Holland) at a flow rate of 10 96% ethanol (Panreacs, Barcelona, Spain) were used L/hour. Light was provided by fluorescent tubes as anesthetic during handling. Fish was measured (Sera blue sky, 36W, 12000º K, Germany) and the from the tip of the snout to the tip of the tail, dried photoperiod was 14 h light: 10 h dark. Temperature on blotting paper and weighed on an electric balance (ºC) and oxygen level (mg L-1) were measured daily (Mettler Toledo, AG204, Greifensee, Switzerland) using a multiparameter instrument (YSI before being returned to the tank. In each sampling Incorporated, Model 55, USA). Measurements of were controlled 10% fish of each tank, except in the other physical parameters as salinity (ppt, initial and final sampling where all individuals were refractometer, Shibuya, Model S-10, Japan), and pH measured and weighted. (Cyberscan PCD-5500, Eutech, USA), were carried out also daily. Nitrogen compounds were measured Concerning statistical analysis, normality twice a week (Seachem Multitest, USA). Plastic and homogeneity of the variable were tested using meshes were added to the tanks in order to provide Kolmogorov Smirnov and Levene’s test holdfast elements for juveniles. respectively [21]. One way ANOVA [21] was used to determine the significance between data and Four experimental diets were tested by triplicate Student-Newman-Keuls test for comparisons for the greater pipefish (S. acus) first feeding: between groups. All results were processed using SPSS statistical software system version 16.0.1. T1: Non-enriched Artemia metanauplii (SPSS, Chicago, IL, USA, 1999)

6 T2: Tetraselmis suecica (0,5x10 cellules/ml) 3 Results enriched Artemia metanauplii. The average temperature during the experiment was T3: Isochrysis galvana (10 6 cellules/ml) enriched 20.69± 0.54 °C, oxygen levels were 6.90± 0.4 ppm Artemia metanauplii and pH 6.90± 0.4, along the experiment. Concerning chemical parameters, ammonia and T4: DHA protein SELCO (0.5 g SELCO/l INVE) nitrites levels were always under 0.2 and 0.02 ppm enriched Artemia metanauplii. during the trials. Culture of microalgae I. galvana and T. Spawning episodes occurred during August suecica were provided by the aquarium laboratory and September. The number of individuals per batch and maintained at 20 ºC in 6 L Pyrex® flasks. was between 245 and 429 for S. acus; for S. thyple Culture evolution was daily checked by a 153 individuals were collected in the only spawning photometric evaluation testing turbidity with a episode during the experimental period. Since the portable photometer (Mod.PF-11; Macherey-Nagel; moment that pipefish are expelled from the paternal Durew, Germany) and individual counts using pouch hunting behaviour has been observed and it haemocytometer (Mod. Neubauer, Germany). can be described as active and voracious. Prey ingestion in S. acus for all treatments is shown in Pipefish were fed on Artemia once a day at Figure 1. 10:00 a.m., at a concentration of 10 metanauplii/ml, according to the aquarium practices. Before daily feed administration, faeces were removed from the bottom of the tanks and remaining Artemia carefully counted in order to determine prey ingestion. Each day, mortality for each treatment was also controlled.

During the experimental period, pipefish were sampled for length and weight at 1, 14, 22 and 28 days after birth (DAB). Pipefish were removed from the tanks before feeding and placed in a tray containing the anaesthetic solution in order to be sampled. Concentrations of 10 ppm of natural clove

ISBN: 978-1-61804-280-4 53 Advances in Environmental Science and Energy Planning

Figure 2. Ingestion of S. acus and S. typhle for DHA protein SELCO (0.5 g SELCO/l INVE) enriched Artemia during the first month of life.

Regarding the use of clove oil as anesthetics in these species no references has been found. All animals reached numb stage with movement (phase II, [22]) during the first 5 min after anesthetic administration at the selected dose. Finally, animals recovered normal activity within 10 min after introduction into the tank. No side-effect after this handling has been observed during the trial.

In reference of survival, no differences were Figure 1. Artemia ingestion of S. acus for the found between treatments (P≤0.01), survival was up different treatments during the first month of life. to 98 % for both species. On average S. acus measured 3.31 cm ±0.261 and weighted 31.03 mg T1: Non-enriched Artemia metanauplii ±0.002 at birth (Table 1), no differences were 6 observed in initial lengths and weights from T2: Tetraselmis suecica (0.5x10 cellules/ml) different batches (P≤0.01). On average S. thyple enriched Artemia metanauplii. measured 3.40 cm ±0.187 and weighted 17 mg ±0.002 at birth (Table 1). T3: Isochrysis galvana (10 6 cellules/ml) enriched Artemia metanauplii Table 1. Growth parameters (Weight and Length, mg and cm, respectively) of S. acus (T1,T2, T3, and T4 : DHA protein SELCO (0.5 g SELCO/l INVE) T4) and S. typhle (T4) fed on different diets. enriched Artemia metanauplii.

It can be observed an increasing ingestion until 20 DAB, when it seems to stabilize around 10 metanauplii/ml. No differences in prey ingestion Day 1 14 22 28 were observed between diets (P˂ 0.05, Fig.1). When comparing prey ingestion between both species for W 31±2.0 46±5.1 75±3.5 95±16.3 T4 diet no differences were observed (P˂ 0.05, T1 Fig.2). L 3.48±0.08 4.60±0.23 5.30±0.39 5.50±0.30

W 31 ±2.0 45±4 75±5 92±18 T2 L 3.48±0.08 4.38±0.24 5.28±0.35 5.52±0.36

W 31.2±1.5 48±6.2 74±3.5 79±16.2b T3 L 3.31±0.22 4.74±0.32 5.48±0.22a 5.42±0.50b

W 30.9±1 57±4a 89±1a 104±12a T4 L 3.14±0.38 4.88±0.08 6.02±0.08a 6.15±0.22a

W 17±3 55±3 117±3 146±5 T4 L 3.40±0.19 5.29±0.07 6.48±0.22 7.24±0.13

In the same column, means with letter denote significant differences.

ISBN: 978-1-61804-280-4 54 Advances in Environmental Science and Energy Planning

W (Weight); L (Length). ppm has been successfully used, denoting the special sensitivity of these species to this anesthetic. T1: Non-enriched Artemia metanauplii The use of this anesthetic for these species showed a high efficacy at low doses, cost-efficiency, non- 6 T2: Tetraselmis suecica (0.5x10 cellules/ml) toxicity, handler safety and non negative side enriched Artemia metanauplii. effects, highlighting its suitability for aquaria use.

6 T3: Isochrysis galvana (10 cellules/ml) enriched No significant difference was recorded in Artemia metanauplii initial lengths and weights of animals from different batches (P ≤ 0.001), differing from results obtained T4: DHA protein SELCO (0.5 g SELCO/l INVE) by Payne et al. [8], surely due to differences in enriched Artemia metanauplii. genitors performance.

At the end of the experiment, S. acus The newborn showed benthic behavior, juveniles fed on Selco are significantly bigger remaining close to the bottom, as well was observed comparing with juveniles fed on other treatments, by other authors [28]. The observed feeding action which didn’t show any differences between them. (dorsal rotation of head and quickly mouth This difference was observed from 14 DAB, until movement through the water) was consistent with the end of experimental period (Table 1). Besides, other authors’ observations [8]. This feeding juveniles fed on Selco were heavier comparing with behaviour observed immediately after emerging juveniles fed on other treatments from day 14 (Table from the brood pouch has been also referred in S. 1). No differences were observed in juveniles fed argus [8], as well as in other syngnathids [17]. This with non-enriched and T. suecica - enriched fact also explained the gut colour of animals after Artemia, however juveniles fed on metanauplii feeding episodes in all treatments. However, it is enriched on I. galvana showed a lower weight from possible that live preys are eaten but their nutrients 22 DAB (Table 1). not completely assimilated by animals in the first days [29], [30]. Regarding the observed prey 4 Discussion ingestion, the common practices at aquaria seems to cover the newborn needs. In the first week it would The physical-chemical parameters showed be considered a reduction on the daily ration in stability during the trial and their values may be order to reduce costs; however, the food density considerate in a normal range. In wild, spawning decrease will produce an increasing effort of periods occur mainly between May and July [23], in catching preys, affecting survival rate. aquaria usually occur between August and September. The success of larval rearing is greatly influenced by first feeding regimes and the On average, number of newborn for batches nutritional quality of starter diets [18]. The high obtained in this study were lower than reported from results obtained in survival rates (up to 98%), wild conditions [24] surely due to captivity showed that Artemia metanauplii administrated in conditions, mainly broodstock diets has been these conditions is appropriated as first feeding for reported as a factor that can have great influence on Syngnathus, as well as has been demonstrated for brood quantity [25] . other sygnathids species as Hippocampus hippocampus [17] and H. reidi [26]. This efficient Concerning the use of clove oil, the results showed first feeding protocol using Artemia simplify for the first time their effectiveness for these species pipefish production methods in aquaria. at a concentration of 10 ppm without negative side effects for animals at this temperature. Clove oil Besides, no significantly differences has been also found useful in other species of between treatments were observed until 14 DAB marine fish [22] even syngnathid species [17], [26]. (Table 1). It must be taking into account the stage of This anesthetic has been used at higher development reached by newborn of these species concentrations (40 ppm) for other marine fish (e.g. when they are released from paternal pouch and the amberjack, sea-bream, European sea-bass, and other first feeding take place. This is not the case for most species [22] even for other syngnathid species (20 marine fish which are at an earlier stage of ppm, H. reidi, [27]; 25 ppm, H. hippocampus, [17]). development when first feeding occurs. These However, for S. acus and S. thyple, a dose of 10 results suggested the possibility to use unenriched

ISBN: 978-1-61804-280-4 55 Advances in Environmental Science and Energy Planning

Artemia during the first 2 weeks and enriched prey, [4] Martin-Smith KM, Vincent ACJ. 2006. after that point. This breeding protocol could be a Exploitation and trade in Australian useful option to decrease production costs in this seahorses, pipehorses, sea dragons and species culture, as well as been recommended for pipefishes (Family Syngnathidae). Oryx 40: other species [26]. 141–151. [5] Shokri, M.R., Gladstonea, W., Jelbar, J. However, after 14 DAB the observed 2009.The effectiveness of seahorses and differences between the treatments showed the pipefish (Pisces: Syngnathidae) as a major role play by Artemia enrichment in this flagship group to evaluate the conservation period. It has been largely recognized that Artemia value of estuarine beds. Aquatic have nutritional deficiencies for marine species, Conserv: Mar. Freshw. Ecosyst. 19, 588– particularly in HUFA (e.g., docosahexaenoic acid 595. and eicosapentaenoico acid). Enrichment of this live [6] Vincent, A.C.J. 1996. The International food with HUFA-rich lipid emulsions, or several Trade in Seahorses. Cambridge: TRAFFIC species or microalgae can provide the dietary International. requirement for fish larvae. In this case, SELCO– [7] Vincent, A.C.J., Foster, S.J., Koldewey, enrichment provides the essential nutrients to obtain H.J. 2011. Conservation and management of the better growth in this period for this species. seahorses and other Syngnathidae. Journal Despite the high concentrations of DHA found in I. of Fish Biology 78, 1681–1724. galbana in comparison with other microalgae [8] Payne, M.F., Rippingale, R.J., Longmoreet, species [31], the worst ongrowing was observed for R.B. 1998. Growth and survival of juvenile this treatment. The high development of pipefish pipefish (Stigmatopora argus) fed live after birth showed by functional eyes and predation copepods with high and low HUFA content. activity suggest that the development of the nervous Aquaculture 167, 237–245. system during which DHA must be provided ([32] [9] Roberts, C.M., Hawkins, J.P. 1999. [33]) passes in an early stage when the fish are Extinction risk in the sea. Trends Ecol. protected within the paternal brood pouch [8]. No Evol., 14(6) 241-246. differences were observed between T1 and T2 (non- [10] Browne, R.K., Baker, J.L., Connolly, R.M. enriched and Tetraselmis enriched Artemia, 2008. Syngnathids: sea dragons, seahorses, respectively) during the experimental period, both and pipefish of Gulf St. Vincent, in: length and weight. These results suggest the Shepherd, S., Bryars, S., Kirkegaard, I., possibility to remove the use of microalgae for Harbison, P., and J.T. Jennings, (Eds.) Artemia enrichment of these species with an Natural History of Gulf St. Vincent. Royal important reduction in operational costs. Society of South Australia, Inc., South Nevertheless, other experiments must be performed Australia, pp.281-305. to test different enrichment products at different . dose or enrichment time in order to optimize its use [11] Gongzhao, X.1985. Fish mariculture and the pipefish ongrowing during the first month of studies in China. ICLARM Newsletter 8, 5– life. 6 [12] Prein, M. 1995. Aquaculture potential of References: seahorses and pipefishes. Naga, 18(1), 20- 21. [1] Espino, F., F. Tuya, I. Blanch y R. J. [13] Molina L., Segade, A. 2011. Aquaculture Haroun, 2008. Los sebadales en Canarias. as a potential support of marine aquarium Praderas de fanerógamas marinas. BIOGES, fish trade sustainability, in: Proceedings of Universidad de Las Palmas de Gran the Third International conference on Canaria, 68 pp... Management of Natural Resources, [2] Bauchot, M.L., Pras, A. 1987. Guía de los Sustainable Development and Ecological peces de mar de España y Europa. Hazards. WitpressTransactions on Ecology [3] Dawson, C.E. 1986. Syngnathidae, in P.J.P. and the Environment (Eds.). Southampton, Whitehead, M.-L Bauchot, J.-C. Hureau, J. pp:15-25. Nielsen and E. Tortonese (eds.). Fishes of [14] Calado, R. Narciso, L. Araújo, R., Lin, J. the north-eastern Atlantic and the 2003. Overview of marine ornamental Mediterranean. UNESCO, Paris. Vol. 2. pp. Aquaculture, in: Marine Ornamental 628-639. species: Collection, Culture &

ISBN: 978-1-61804-280-4 56 Advances in Environmental Science and Energy Planning

Conservation. J.C. Cato and C.L. Brown reproduction. J. Mar. Biol. Assoc. U.K (Eds.) Iowa State Press, pp.219-230. 85:1235-1241. [15] Taskavac, E., Gürkan, S. Sever, T.M., [25] Lin Q, Lu J, Gao Y, Shen L, Cai J, Luo J. Akalin, S., Özaydin, O. 2010. Gut contents 2006. The effect of temperature on gonad, and feeding habits of the great pipefish, embryonic development and survival rate of Syngnathus acus Linnaeus, 1758, in Izmir juvenile seahorses, Hippocampus kuda Bay (Aegean Sea, Turkey). Zoology in the Bleeker. Aquaculture 254:701-713. Middle East 50, 75-82. [26] García-Manchón, J. 2012. Aportaciones al [16] Ryer, C.H., Orth, R.J. 1987. Feeding conocimiento de la reproducción y cultivo Ecology of the Northern Pipefish, larvario de Hippocampus reidi en Syngnathus fuscus, in a Seagrass condiciones de cautividad. MSc Thesis Community of the Lower Chesapeake Bay. Universidad de Las Palmas de Gran Estuaries 10 (4), 330-336. Canaria. [17] Otero-Ferrer F., Molina L., Socorro J., [27] García-Manchón, J., Otero-Ferrer, F., Herrera R., Fernández-Palacios H., Segade, A., Novelli, B., Socorro, J., Molina, Izquierdo, MS. 2010. Effects of first feeding L. 2011 Anaesthetic protocol for biometric on survival and growth of short-snouted study in Hippocampus reidi (Ginsburg, seahorse juveniles, Hippocampus 1933) juveniles, in: Proceedings of the hippocampus, (Linnaeus, 1758). Aquac. Aquaculture Europe 2011 electronic Res. 41:8-19. publication. Rhodes. [28] Silva, K., Monteiro, N.M., Almada, V.C., [18] Izquierdo MS, Tandler A, Salhi M, Vieira, M.N. 2006. Development and early Kolkovsky S. 2001. Influence of dietary life history behaviour of aquarium reared polar lipids’ quantity and quality on Syngnathus acus (Pisces: Syngnathidae). J. ingestion and assimilation of labelled fatty Mar. Biol. Assoc. U.K. 86:1469-1472. acids by larval gilthead seabream. Aquacult. [29] Warland, T. 2003. Seahorses. How to care Nutr.7:153-160. for your Seahorses in the Marine Aquarium. Port Lincoln: South Australian seahorse [19] Sargent J.R., Bell G, McEvoy. marine services. L.A.Tocher, D. and Esteve, A. 1999 [30] Novelli, B. 2012. Aplicación de técnicas Recent developments in the essential fatty histológicas y osteológicas al estudio de la acid nutrition of fish. Aquaculture ontogenia en Hippocampus sp. MSc thesis 177:191-199. Universidad de Las Palmas de Gran [20] Ross,G. and Ross, B. Anaesthetic and Canaria. sedative techniques for aquatic animals.2nd [31] Conceiçao, L.E.C., Yúfera, M., Makridis, edition Blackwell Science, Oxford, 159 pp. P., Morais, S., Dinis, M. T. 2009 Live feeds [21] Zar, J.H. 1996. Biostatistical analysis, 3rd for early stages of fish rearing. Aquaculture ed., Upper Saddle River, NJ: Prentice–578 Research 2009, 1-28. Hall [32] Mourente, G., Tocher, D.R., Sargent, J.R., [22] García-Gómez A, Gándara F., Raja, T. 1991. Specific accumulation of 2002. Utilización del aceite de clavo, docosahexaenoic acid 22:6n-3 in brain Syzygium aromaticum L. (Merr. & Perry), lipids during development of juvenile turbot como anestésico eficaz y económico para Scophthalmus maximus L. Lipids 26, 871– labores rutinarias de manipulación de peces 877. marinos cultivados. Bol. Inst. Esp. [33] Tocher, D.R., Mourente, G., Sargent, J.R., Oceanogr. 18 (1-4):21-23. 1992. Metabolism of (1-14C) [23] 14 Vincent, A.C.J., Berglund, A., docosahexaenoate (22:6n-3), (1- C) Ahnesjö, I. 1995. Reproductive ecology eicosapentaenoate (20:5n-3) and (1-14C) of five pipefish species in one eelgrass lionlenate (18:3n-3) in brain cells from meadow. Environ. Biol. Fish. 44:347- juvenile turbot Scophthalmus maximus. 361 Lipids 27, 494–499. [24] Monteiro, N.M., V.C. Almada and M.N. Vieira, 2005. Implications of different brood pouch structures in Syngnathid

ISBN: 978-1-61804-280-4 57