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A Microalga Is Better Than a Commercial Lipid Emulsion at Enhancing Live Feeds for an Ornamental Marine Fish Larva

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A Microalga Is Better Than a Commercial Lipid Emulsion at Enhancing Live Feeds for an Ornamental Marine Fish Larva Please do not remove this page A microalga is better than a commercial lipid emulsion at enhancing live feeds for an ornamental marine fish larva Basford, Alexander J; Mos, Benjamin; Francis, David S; et al. https://researchportal.scu.edu.au/discovery/delivery/61SCU_INST:ResearchRepository/1268876610002368?l#1368876590002368 Basford, A. J., Mos, B., Francis, D. S., Turchini, G. M., White, C. A., & Dworjanyn, S. (2020). A microalga is better than a commercial lipid emulsion at enhancing live feeds for an ornamental marine fish larva. Aquaculture, 523. https://doi.org/10.1016/j.aquaculture.2020.735203 Document Version: Accepted Published Version: https://doi.org/10.1016/j.aquaculture.2020.735203 Southern Cross University Research Portal: https://researchportal.scu.edu.au/discovery/search?vid=61SCU_INST:ResearchRepository [email protected] CC BY-NC-ND V4.0 Embargo Downloaded On 2021/09/27 15:03:21 +1000 Please do not remove this page 1 A microalga is better than a commercial lipid emulsion at enhancing live feeds for an 2 ornamental marine fish larva 3 4 Alexander J. Basford ab*, Benjamin Mos a, David S. Francis c, Giovanni M. Turchini c, 5 Camille A. White d, Symon Dworjanyn a 6 7 a National Marine Science Centre, Southern Cross University, Coffs Harbour 2450, Australia 8 b Darwin Aquaculture Centre, Department of Primary Industry and Resources, Northern 9 Territory Government, Channel Island 0822 10 c Deakin University, School of Life and Environmental Sciences, Geelong 3220, Australia 11 d Oceans and Atmosphere, Food Nutrition and Bio-based Products, Commonwealth Scientific 12 and Industrial Research Organization, Hobart 7000, Australia 13 * Corresponding author: [email protected] (A. J. Basford) 14 15 Highlights 16 • Larval Amphiprion latezonatus grew faster and had a higher survival rate when fed 17 live feeds enhanced with the microalga Proteomonas sulcata compared to live feeds 18 enriched using a commercial lipid emulsion. 19 • A. latezonatus fed live feeds enriched with a commercial lipid emulsion had the 20 highest DHA content, but this did not correlate with survival or growth. 21 • DPAn-6 and the ratio of DPAn-6 to DHA in A. latezonatus tissues was positively 22 correlated with larval survival and growth. 23 • High levels of dietary DHA may be less important for some marine fish larvae than 24 previously thought. 25 26 Key words: Amphiprion, anemonefish, cryptomonad, larval rearing, phospholipids, fatty 27 acids, 22:6n-3, 22:5n-6 28 Abstract 29 High mortality during larval rearing is a persistent bottleneck in finfish aquaculture 30 and is often caused by inadequate nutrition. Nutritional deficiencies in live feeds are usually 31 overcome by enrichment with commercial lipid emulsions; however, enriching live feeds 32 using microalgae may be more effective. This study investigated the efficacy of the microalga 33 Proteomonas sulcata as a live feed enhancement for rearing larvae of the wide-band 34 anemonefish, Amphiprion latezonatus. We found A. latezonatus larvae fed live feeds 35 enhanced with P. sulcata had higher survival and better growth at 7 days post hatch (dph) 36 compared to fish fed live feeds enriched with a commercial lipid emulsion or left unenriched. 37 At 14 dph, A. latezonatus initially fed rotifers cultured on P. sulcata for several generations 38 had the highest survival overall. These results may be due to the high phospholipid content in 39 rotifers enhanced on P. sulcata compared to other diets. Survival, length, depth, and eye 40 diameter of 7 dph A. latezonatus was also positively correlated with omega-6 41 docosapentaenoic acid (DPAn-6, 22:5n-6) and high ratios of DPAn-6 to docosahexaenoic 42 acid (DHA, 22:6n-3) in their tissues, highlighting the need to better understand the role of 43 DPAn-6 in larval fish ontogeny. Surprisingly, given their high survival and growth, A. 44 latezonatus larvae fed P. sulcata enhanced live feeds had lower levels of DHA compared to 45 the more poorly performing fish fed the commercial lipid emulsion enriched live feeds; this 46 challenges the paradigm of a positive correlation between DHA and growth and survival that 47 is documented in many other marine fish larvae. This study demonstrates the benefits of 48 using P. sulcata to enhance the nutritional quality of live feeds and highlights the need for a 49 better understanding of the role of n-6 long-chain polyunsaturated fatty acids in the early 50 development of larval fish. 51 1. Introduction 52 The majority of marine fish sold in the ornamental aquarium trade are wild caught 53 from coral reefs (Rhyne et al., 2014). This harvest is an additional stressor for one of the most 54 threatened ecotypes on the planet, but could be alleviated if more marine ornamental fish 55 were produced in closed cycle aquaculture (Ban et al., 2014). The small proportion of 56 cultured finfish in the ornamental trade is largely due to the difficulty in rearing larvae 57 (Calado et al., 2017), similar to the persistent bottleneck caused by high mortality of fish 58 larvae grown for food (Hamre et al., 2013; Rønnestad et al., 2013). 59 An important contributor to mortality during larval rearing is inadequate nutrition in 60 live feeds (Hamre et al., 2013). Rotifers, Brachionus spp., and brine shrimp, Artemia spp., are 61 the most common live feeds used in aquaculture, but contain insufficient amounts of essential 62 nutrients required by fish larvae such as long chain (≥20 chain length) polyunsaturated fatty 63 acids (LC-PUFA) and phospholipids (Rainuzzo et al., 1997; Sorgeloos et al., 2001; Hamre et 64 al., 2008; Hamre et al., 2013). Lipids are the most energy dense nutrients available to marine 65 fish, and larvae require high levels of specific LC-PUFA such as arachidonic acid (ARA, 66 20:4n-6), omega-6 docosapentaenoic acid (DPAn-6, 22:5n-6), eicosapentaenoic acid (EPA, 67 20:5n-3), and docosahexaenoic acid (DHA, 22:6n-3), due to their inability to synthesise these 68 fatty acids from more commonly available, shorter-chain and less unsaturated, precursors 69 (Parrish et al., 2007; Glencross, 2009; Tocher 2010; Izquierdo and Koven, 2011). Marine fish 70 larvae also require dietary LC-PUFA in specific ratios that vary among species (Glencross 71 2009). It is generally accepted that more dietary DHA should be provided than EPA and 72 ARA, although there is less clarity about what DHA to DPAn-6 ratios are ideal (Sargent et 73 al., 1997). DHA is preferentially conserved over other LC-PUFA and plays an essential role 74 in larval fish survival and development (Watanabe, 1993; Sargent et al., 1999; Hamre et al., 75 2013). 76 In aquaculture, DHA and other essential fatty acids are typically delivered to marine 77 fish larvae by enriching live feeds with lipid emulsions (Dhert et al., 2001; Ma and Qin, 78 2014). Commercially available lipid emulsions are typically rich in n-3 LC-PUFA, 79 particularly DHA, and are used to increase the levels of these nutrients in rotifers and 80 Artemia. However, this is primarily in the form of neutral lipids and triacylglycerols 81 (Coutteau and Mourente, 1997; Li et al., 2015). While live feeds with high neutral lipid 82 content can be nutritious (Estévez and Giménez, 2017), larval fish should ideally receive 83 dietary lipids in the form of polar lipids, such as phospholipids, which are more efficiently 84 assimilated (Olsen et al., 2014). In addition, lipid emulsions are expensive, and live feeds 85 egest gut-loaded lipid emulsions, adversely affecting water quality (Høj et al., 2009; 86 Yamamoto et al., 2009). 87 An alternative to lipid emulsions for enhancing the nutritional content of live feeds is 88 microalgae (Ma and Qin, 2014; Rehberg-Haas et al., 2015a; Ferreira et al., 2018). Live feeds 89 raised on microalgae can incorporate desirable nutrients such as LC-PUFA in the form of 90 biologically available phospholipids (Rainuzzo et al., 1994a; Li and Olsen, 2015; Li et al., 91 2018) and pass these nutrients to their offspring. This can amplify the content of these 92 nutrients in live feeds over generations (Koiso et al., 2009; Li et al., 2015; Sperfeld and 93 Wacker, 2015). Attempts to grow rotifers on microalgae have been hindered by the use of 94 relatively non-motile genera such as Pavlova and Isochrysis, which settle out of culture and 95 degrade water quality (Ferreira et al., 2008; Rehberg-Hass et al., 2015b). Motile microalgae 96 such as cryptomonads remain suspended in the water column and are already used as food for 97 invertebrate larvae (Knuckey et al., 2005; Mos et al., 2011), raising the possibility that these 98 microalgae may be suitable for enhancing the nutritional content of live feeds. 99 This study compared the nutritional quality of live feeds enhanced with the 100 cryptomonad microalga, Proteomonas sulcata, to live feeds enriched with a commercial lipid 101 emulsion. Most studies on improving the nutritional value of live feeds for larval fish have 102 used commercially important food finfish, while similar studies on ornamental species are 103 scarce (Calado et al., 2017). In this study, larvae of the wide-band anemonefish Amphiprion 104 latezonatus were used as a model ornamental fish species. A. latezonatus is a rare and 105 valuable species in the aquarium trade due to restricted wild collection from its small home 106 range (Scott et al., 2011). To gauge the efficacy of mutigenerational and short term 107 enhancement of live feeds with microalgae, we compared the survival and growth of A. 108 latezonatus larvae fed rotifers that were: 1. Cultured on P. sulcata for several generations; 2. 109 Enriched with P. sulcata overnight; 3. Enriched with a commercial lipid emulsion (Selco 110 S.presso); or 4.
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