Journal of the World Aquaculture Society Demonstr ation That Feeds Containing <1% Fish Meal Can Support Growout Of Large Juvenile Red Drum, Sciaenops ocellatus, And Reduce Nutrient Waste For Peer Review Journal: Journal of the World Aquaculture Society Manuscript ID JWAS-16-114.R2 Manuscript Type: Applied Studies Date Submitted by the Author: n/a Complete List of Authors: Denson, Michael; Marine Resources Research Institute, Department of Natural Resources Sandifer, Paul; College of Charleston, School of Sciences and Mathematics; Hollings Marine Laboratory, College of Charleston Leffler, John; South Carolina Department of Natural Resources, Marine Resources Research Institute Yost, Justin; Marine Resources Research Institute, Department of Natural Resources Bearden, Daniel; National Institute of Standards and Technology Zeigler, Tom ; Zeigler Bros. Inc. fish meal replacement, alternative protein sources, reduced pollution, Keywords: enzyme additives, red drum Journal of the World Aquaculture Society Page 1 of 38 Journal of the World Aquaculture Society 1 1 2 3 1 Demonstration That Feeds Containing <1% Fish Meal Can Support Growout Of Large Juvenile 4 5 6 2 Red Drum, Sciaenops ocellatus, And Reduce Nutrient Waste 7 8 9 3 10 11 12 4 Michael R. Denson, Marine Resources Research Institute, South Carolina Department of Natural 13 14 15 5 Resources, 217 Fort Johnson Road, Charleston, SC 29412 USA 16 17 18 6 Paul A. Sandifer, CollegeFor of Charleston, Peer Hollings Review Marine Laboratory, 331 Fort Johnson Road, 19 20 7 Charleston, SC 29412 USA 21 22 23 8 John W. Leffler, Marine Resources Research Institute, South Carolina Department of Natural 24 25 26 9 Resources, 217 Fort Johnson Road, Charleston, SC 29412 USA 27 28 29 10 Justin Yost, Marine Resources Research Institute, South Carolina Department of Natural 30 31 11 Resources, 217 Fort Johnson Road, Charleston, SC 29412 USA 32 33 34 35 12 Daniel W. Bearden, National Institute of Standards and Technology, Hollings Marine 36 37 13 Laboratory, 331 Fort Johnson Road, Charleston, SC 29412 USA 38 39 40 14 Thomas R. Zeigler, Zeigler Bros., Inc., 400 Gardners Station Road, Gardners, PA 17324. 41 42 43 15 Corresponding Author: Paul A. Sandifer, College of Charleston, Hollings Marine Laboratory, 44 45 46 16 331 Fort Johnson Road, Charleston, SC 29412 USA. [email protected] 47 48 17 49 50 51 18 52 53 54 55 56 57 58 59 60 Journal of the World Aquaculture Society Journal of the World Aquaculture Society Page 2 of 38 2 1 2 3 19 Abstract 4 5 6 7 20 We conducted a 16-week feeding trial with large juvenile red drum, Sciaenops 8 9 21 ocellatus. Four diets were randomly assigned to six replicate tanks per treatment. Three 10 11 22 isonitrogenous (~44.5% protein) and isolipidic (~14.1% lipid) extruded diets were 12 13 14 23 formulated to compare a fish meal-based diet with diets using alternative protein sources. 15 16 24 Diet 1 contained 19.60% fish meal and 21.42% poultry meals as primary protein sources. 17 18 Two alternative dietsFor were formulated Peer reducing Review the fish meal to 0.61% by substituting 19 25 20 21 26 poultry meals (33.85%) and soybean protein concentrate (11.55% in Diet 2 and 11.70% in 22 23 27 Diet 3. Diet 3 also included Allzyme Vegpro ® and Allzyme ® SSF at 0.04%. Diet 4, a 24 25 26 28 “natural” diet consisting of chopped cigar minnows, squid, and shrimp, was used as a 27 28 29 positive control to compare growth rates of formulated feeds to near maximum growth 29 30 30 under these culture conditions. We found that reducing the amount of fish meal to <1% by 31 32 33 31 using alternative protein sources did not affect the growth rate, survival, or health of red 34 35 32 drum but improved assimilation of Phosphorus, reduced potential release of P to the 36 37 33 environment, and significantly lowered the amount of feeder fish needed in feed. The 38 39 40 34 control diet identified performance benchmarks for future feeds development work . 41 42 43 35 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Journal of the World Aquaculture Society Page 3 of 38 Journal of the World Aquaculture Society 3 1 2 3 36 4 5 37 The necessity for aquaculture to expand to provide animal protein and other products for 6 7 38 humans and to help spare fishery stocks from overexploitation has been emphasized many times 8 9 39 (e.g., Bardach et al. 1974; Pillay 1978; Sandifer 1979). Aquaculture has been growing at 10 11 12 40 sustained rapid rates for many years and now surpasses capture fisheries in the amount of 13 14 41 seafood produced for human consumption (FAO 2016). Concomitant with rapid growth has 15 16 42 come recognition of the need to eliminate or at least substantially reduce aquaculture’s reliance 17 18 For Peer Review 19 43 on fish meal and oil as feed ingredients and to decrease aquaculture’s contribution to nutrient 20 21 44 pollution of receiving waters (Naylor et al. 2009; Tacon and Metian 2008; Browdy and 22 23 24 45 Hargreaves 2009; Price and Morris 2013) 25 26 27 46 Red drum, Sciaenops ocellatus, is an important marine aquaculture species because of its 28 29 47 fast growth rate, wide environmental tolerances and market appeal (Sandifer et al. 1993; Faulk 30 31 48 2005). The cost of feed remains the single greatest cost for production of finfish, and fish meal is 32 33 34 49 often the most expensive component of fish feeds (Rana et al. 2009). Over the last decade, 35 36 50 numerous studies have been conducted to identify and evaluate alternative protein sources for 37 38 39 51 aquafeeds for a variety of freshwater and marine fish and crustaceans, including soybean and 40 41 52 other plant meals, blood meal, poultry meal and other animal processing by-products (Hardy 42 43 53 2000; Rawles and Gatlin 2000; Emre et al. 2003; Erturk and Sevgili 2003; Hernandez et al. 2004; 44 45 46 54 Davis et al. 2005; Gaylord and Rawles 2005; Brinker and Reiter 2011). While a number of 47 48 55 studies have examined the potential to utilize soybean meal and oil and other feed ingredients to 49 50 56 partially replace fish meal in practical feeds for red drum (e.g., Reigh and Ellis 1992, Davis et al. 51 52 53 57 1995; Gaylord and Gatlin 1996; Meilahn et al. 1996; McGoogan and Gatlin 1997, Kureshy et al. 54 55 58 2000, Rossi et al, 2013, 2015, Minjarez-Osorio et al. 2016), complete elimination of fish meal in 56 57 58 59 commercial diets has not yet been achieved. 59 60 Journal of the World Aquaculture Society Journal of the World Aquaculture Society Page 4 of 38 4 1 2 3 60 Two main end-products of intensive modern finfish aquaculture are nitrogen (N) wastes 4 5 6 61 from deamination of proteins and indigestible phytate-bound phosphorus (P) (USEPA 2005; 7 8 62 Debnath et al. 2008). These by-products can have negative effects on fish health and growth 9 10 11 63 within production facilities as well as ecosystem impacts such as eutrophication (Goldburg and 12 13 64 Naylor 2005; Hall et al. 2011) that may lead to hypoxia and/or harmful algal blooms (Anderson 14 15 65 et al. 2008; Heisler et al. 2008). Improvements in prepared diets to mitigate these negative 16 17 18 66 impacts will be a leadingFor factor in Peer the long-term sReviewustainability of aquaculture (USCOP 2004; 19 20 67 Rust et al. 2011). 21 22 23 68 Numerous products have been tested to improve P utilization, feed conversion ratios 24 25 26 69 (FCR), and protein uptake and retention efficiency (Jackson et al. 1996; Li and Robinson 1997; 27 28 70 Van Weerd et al. 1999; Hardy 2000; Cao et al. 2007; Buentello et al. 2010, Refstie et al. 2010). 29 30 71 Compounds that may reduce the pollution stream include new forms of phytase (i.e. enzymes 31 32 33 72 that release P from phytates in plants and make it available to consuming animals), acidifiers, 34 35 73 pre- and probiotics, and essential oils (Debnath et al. 2005; Fox et al. 2006). While some of these 36 37 74 components have been adopted by the aquaculture feed industry, additional research is needed, 38 39 40 75 especially for carnivorous marine species (Buentello et al. 2010). 41 42 43 76 The present study was undertaken to: (1) determine the efficacy of using soybean and 44 45 77 poultry by-product meals to replace nearly all of the fish meal in extruded prepared diets for red 46 47 48 78 drum under grow-out conditions (~250 g to ~500 g); (2) evaluate the potential for reducing N 49 50 79 and P waste loading from red drum aquaculture via such substitutions, with and without enzyme 51 52 80 additives; and (3) to use Nuclear Magnetic Resonance (NMR)-based metabolomics to measure 53 54 55 81 metabolic changes in various fish tissues in response to the diets presented. This paper focuses 56 57 82 on results from objectives 1 and 2; results from the NMR studies will be the subject of a separate 58 59 60 Journal of the World Aquaculture Society Page 5 of 38 Journal of the World Aquaculture Society 5 1 2 3 83 manuscript. Although most nutritional studies of this type use very small fish (2-10 g in weight) 4 5 6 84 and trials of short duration (30-60 days), we chose to use larger red drum that more accurately 7 8 85 reflect dietary needs and fish metabolism during grow-out to harvestable size.