Aquaculture 465 (2016) 5359

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Aquaculture

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Use of dry hydrolysate from squid and scallop product supplement in

plant based practical diets for Pacic white shrimp Litopenaeus vannamei
Yangen Zhou a, Ramasamy Thirumurugan b, Qingkui Wang c, Chong M. Lee d, D. Allen Davis e,
a
Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, 5 Yushan Road, Qingdao 266003, China
b
Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, India
c
Tianjin Key Laboratory of Aqua-ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, 22 Jinjing Rd, Tianjin 300384, China
d
Food Science and Nutrition Research Center, Department of Nutrition and Food Sciences, University of Rhode Island, 530 Liberty Lane, West Kingston, RI 02892, USA
e
School of Fisheries, Aquaculture, and Aquatic Sciences, Auburn University, Auburn, AL 36849-54119, USA

a r t i c l e i n f o a b s t r a c t

Article history: A series of trials were designed to evaluate hydrolysates from waste streams of squid and scallop processing cen-
Received 28 April 2016 ters for the use in practical shrimp feeds. Towards this goal, three growth trials and a consumption trial were con-
Received in revised form 16 August 2016 ducted to evaluate the use of dry hydrolysate from squid and scallop waste as specialty ingredients in commercial
Accepted 19 August 2016
type feed formulations for Pacic white shrimp Litopenaeus vannamei. In the three six-week growth trials, the
Available online 25 August 2016
basal diet (35% crude protein, 8% lipid) consisted primarily of soybean meal (SBM), corn protein concentrate,
Keywords:
corn starch, and whole wheat. In Trial 1, the basal diet was supplemented with 3%, 6%, and 9% from squid hydro-
Dry scallop hydrolysate lysate (Squ) or scallop hydrolysate (Sca) as well as two treatments using SBM which was impregnated with the
Dry squid hydrolysate hydrolysate prior to use. In Trial 2, the basal diet was supplemented with 3% Squ or Sca, shrimp but the shrimp
Growth were offered diets in slight excess to allow for possible increases in consumption of feeds. In Trial 3, Squ and
Litopenaeus vannamei Squid meal were compared at 6%. At the end of the three growth trials, no signicant differences were found
Plant based diet in nal biomass, nal mean weight, percent weight gain, feed conversion ratio, and survival. Additionally, a
group of shrimp (3 g initial mean weight) were used to estimate consumption of the feeds over three 1-hour
feedings over three days. Interestingly, the estimated feed intake of the basal diet was signicantly higher than
that of diets supplemented with 3% Squ or Sca. Results of the present study indicate that Squ and Sca as well
as the SBM impregnated with Squ or Sca are good marine ingredients and can be used as feed ingredients. How-
ever, no evidence of improved feed intake or enhancement of growth were observed even though the diets were
plant based.
Statement of relevance: In this study, a series of trials were used to assess the efcacy of dry hydrolysate from
squid and scallop as well as traditional squid meal use in plant based practical diets designed for the Pacic
white shrimp. These ndings will be useful because the feed producers are interested in how specic produce
may improve performance either as a nutrient source or attractant, particularly in diets that do not contain
shmeal. The conclusion provide valuable information that indicates feed manufacturer and researchers can
use these hydrolysate products as feed ingredients in diets of shrimp, but no evidence for use of these products
to improve the shrimp growth performance was observed.
2016 Elsevier B.V. All rights reserved.

1. Introduction ingredients that are viewed as attractants or feeding stimulants has

As commercial feeds transition from sh meal based diets to those of
alternative ingredients, the use of attractants/palatability enhancers
may become more crucial (Nunes et al., 2006). It has been suggested
that high dietary levels of alternative ingredients may reduce palatabil-
ity resulting in a reduction of feed intake and negative impacts on the
animals' performance (Watanabe, 2002; Watanabe et al., 1998). To
tackle the challenge of palatability using plant-based diets, inclusion of
Corresponding author.
E-mail address: davisda@auburn.edu (D.A. Davis).
demonstrated to improve the palatability of low sh meal diets in
some species of shrimp and sh (Chotikachinda et al., 2013; Deng
et al., 2013).
Either freeze-dried hydrolysate meals or extracts of natural organ-
isms like squid, scallop, krill, worms, or mussel could be viewed as at-
tractants to enhance dietary palatability (Akiyama et al., 1984; Hartati
and Briggs, 1993; Hernndez et al., 2011; Kader et al., 2012; Nagel
et al., 2014; Smith et al., 2005). A considerable number of studies have
demonstrated that sh hydrolysate can be used as protein supplements,
attractants and palatability enhancers in aquatic animal feeds (Aksnes
et al., 2006; Anggawati et al., 1990; Bui et al., 2014; Hernndez et al.,

http://dx.doi.org/10.1016/j.aquaculture.2016.08.028
0044-8486/ 2016 Elsevier B.V. All rights reserved.
54 Y. Zhou et al. / Aquaculture 465 (2016) 5359
2011; Murray et al., 2003; Nguyen et al., 2012; Oliva-Teles et al., 1999;
Refstie et al., 2004; Zheng et al., 2012). Some previous studies reported
that squid hydrolysate can be used as a protein source or feed attrac-
tant/stimulant in sh (Lian and Lee, 2003a, 2003b). Crdova-Murueta
and Garca-Carreo (2002) stated that the hydrolyzed protein from
sh or krill is a good supplement in diets of Pacic white shrimp.
As marine products are viewed as good nutrient sources as well as
possible attractants/palatability enhancers, there is interest in recover-
ing waste by products from the processing of marine products. In the
northeast coastal regional of United States, approximate
4500 metric tons of squid byproducts are annually generated by the cur-
rent practice of squid processing, which can reach about 50% of the total
squid weight (Lian et al., 2005). Lian et al. (2005) documented that
squid hydrolysate contained 10.76% protein, 1.78 lipid (11.16%
eicosapentaenoic acid [EPA] and 24.45% docosahexaenoic acid [DHA]
in oil), 1.03% ash, and 87.05% moisture. Recently, Lee and Lian (2010)
and Karayannakidis et al. (2014) reported a commercially feasible bio-
conversion process for making squid hydrolysate has been developed
and improved. Thus, the squid hydrolysate has potential application in-
clude 1) feed ingredient for larval and juvenile sh in aquaculture (Lee,
2004; Lian et al., 2008) 2) pet feed stimulant 3) organic fertilizer (Fetter
et al., 2012).
Albeit there are a number of studies demonstrating potential of pro-
tein sources as attractant and palatability enhancers in puried diets,
there is limited data in practical diets. In particular, the use of marine
byproducts as protein sources and possible attractants and feeding
stimulants for marine shrimp using plant-based diets is limited.
Hence, the objective of this study was to evaluate the effect on growth
performance and estimated feed intake of squid and scallop by-
product meals in plant-based diets for the Pacic white shrimp
Litopenaeus vannamei.
2. Materials and methods
2.1. Preparation of dry hydrolysates from squid and scallop products
Squid processing byproducts (SPB) (heads, viscera, skin, ns and
small tubes) were provided by a local squid processor (Sea Fresh,
North Kingstown, Rhode Island) and scallop viscera (SV) (the remain-
der after collecting adductor muscle) obtained from a scallop harvester
(F/V Captain Robert Fisheries, Narragansett, Rhode Island). The raw ma-
terial handling and hydrolysis were carried out at the Food Science and
Nutrition Research Center-Seafood Lab at the University of Rhode Island
(West Kingston, Rhode Island). SPB was used as is after draining off ex-
cess water, while SV was briey rinsed to remove sand. Each sample
(22.69 kg) was chopped at a high speed for 2 min in a Hobart VCM
mixer (30 L working capacity) and transferred into a 49.4 L stainless
steel vessel. The vessel was placed in a large industrial steam-jacketed
kettle (114 L) lled with water which was heated to bring the temper-
ature of homogenate of SPB or SV to 55 C (Karayannakidis et al., 2014).
Due to the relatively small sample size, direct steam injection (DSI) was
not directly applied to homogenate. For the larger production, we plan
to use DSI for hydrolysis in the future. Hydrolysis was brought about
by endogenous proteases present in the viscera and continued for
1.5 h, followed by pasteurization for 30 min at 75 C. According to vis-
cosity change and SDS-PAGE prole reported by Lian et al. (2005), the
viscosity of squid hydrolysate reached the minimum and most protein
molecules were hydrolyzed into peptides and free amino acids after
1.5 h. Resulting hydrolysates were ltered using a 1.18 mm sieve (No.
16). Filtered hydrolysates (squid and scallop) (about 90% yield) were
shipped to Custom Drying Solutions (Valparaiso, Indiana) and dried
using an industrial scale drum dryer which works on the principle of
plate-coat drying used on a pilot scale in our laboratory. Drum-dried hy-
drolysates were ground and sieved through 1.18 mm sieve for particle
uniformity. These products are referred to as dry squid hydrolysate
(Squ) and dry scallop hydrolysate (Sca).
Solvent extracted soybean meal (SBM) was impregnated with either
wet squid or scallop hydrolysate by mixing and allowing equilibration
for adequate impregnation for 1 h and drying in a cabinet dryer at
70 C to approximately 10% moisture. Resulting impregnated SBM was
ground to ner particle size and sieved through 2 mm screen (No. 10).
Proximate analyses of these ingredients were done and are presented
in Table 1.
2.2. Experimental design and diets
Three trials were designed to evaluate the biological response of
shrimp to squid or scallop hydrolysate in soybean meal diets in terms
of growth and feed intake (Table 2). In three trials, the basal diet
(350 gkg1 crude protein and 80 gkg1 lipid) was primarily composed
of soybean meal (SBM), corn protein concentrate, corn starch, and
whole wheat (Table 3). In trial 1, the basal diet SBM was replaced
with 3%, 6%, or 9% Squ or Sca on isonitrogenous and isolipidic basis
(Table 3). Additionally, the last two diets contained high level (54.5%) of
SBM impregnated with 6% squid hydrolysate (ISq6) or 6% scallop hydro-
lysate (ISc6). In Trial 2, equivalent diets as those used in Trial 1 with 3%
Squ or Sca were developed. In Trial 3, the basal diet SBM was replaced
with 6% Squ or 6% Squid meal. The ingredient composition of test diets
in Trials 1, 2, and 3 are presented in Tables 3, 4, and 5, respectively.
Test diets were produced in the Aquatic Animal Nutrition Laboratory
at the School of Fisheries, Aquaculture and Aquatic Sciences, Auburn
University (Auburn, Alabama) utilizing standard protocol for the labora-
tory production of shrimp feeds described by Zhou et al. (2014). Briey,
all dry ingredients were pre-ground, weighed and mixed, followed by
the addition of sh oil and then boiling water. The diets were pelleted
using a meat grinder with a 2.5-mm die, and dried to about 58% mois-
ture. Dry pellets were then crumbled to appropriate size using #43 Disc
Mill (Glen Mills Inc., Clifton, New Jersey), packed in sealed bags, and
stored in a freezer until used. The experimental diets were analyzed at
Agricultural Experiment Station Chemical Laboratories, University of
Missouri (Columbia, Missouri) for proximate composition.
2.3. Growth trials
Three trials were conducted at E. W. Shell Fisheries Research Station,
Auburn University (Auburn, Alabama). Pacic white shrimp post larvae
(PL), were obtained from Shrimp Improvement Systems (Islamorada,
Florida) and nursed in an indoor recirculating system. PL were fed a
commercial PL feed (40% protein and 9% lipid; Zeigler Bros., Inc.,
Gardners, Pennsylvania) using an automatic feeder for about 1 week,
and then switched to crumbled commercial shrimp feed for about 1 or
2 week (36% protein and 8% lipid; Rangen Inc., Buhl, Idaho).
In Trial 1, the recirculating system consisted of 36 aquaria (80 L,
0.50 0.45 0.35 m) connected to a common reservoir, biological lter,
bead lter, uidized biological lter and circulation pump. Four repli-
cate groups of shrimp (0.66 g initial mean weight, 10 shrimp/tank)
were offered diets using our standard feeding protocol over 6 weeks.
Based on the historic results in our laboratory, daily feed rations were
Table 1
Proximate composition (g/100 g as is) of test ingredients. Soybean meal from the same
batch was used in the diets and for impregnation.
Ingredient Moisture Protein Lipid Ash
SBMa 11.02 46.98 1.44 5.93
Squb 8.16 72.22 10.53 6.03
Scac 7.03 59.18 14.41 8.09
ISqd 7.41 49.02 1.58 6.52
ISce 6.45 47.27 3.24 7.50
a
SBM: Solvent extracted soybean meal, Bunge, Alabama.
b
Squ: Dry squid hydrolysate.
c
Sca: Dry scallop hydrolysate.
d
Isq: Impregnated soybean meal with squid hydrolysate.
e
Isc: Impregnated soybean meal with scallop hydrolysate.
 Y. Zhou et al. / Aquaculture 465 (2016) 5359 55

Table 2
Supplemental sources and levels of squid and scallop products in plant based diets used in three trials to evaluate the growth performance of Pacic white shrimp Litopenaeus vannamei.

Trial Trial type Basal diet Supplemental sources Supplemental levels Period

1 Growth trial 56.80% SBM Dry squid hydrolysate 3%, 6%, 9% 6 weeks
Dry scallop hydrolysate 3%, 6%, 9%
Impregnated SBM with 6% squid hydrolysate 56.8%
Impregnated SBM with 6% scallop hydrolysate 56.8%
2a Growth trial 54.10% SBM Dry squid hydrolysate 3% 6 weeks
Dry scallop hydrolysate 3%
2b Estimated feed intake 54.10% SBM Dry squid hydrolysate 3% 3 days
Dry scallop hydrolysate 3%
3 Growth trial 54.10% SBM Dry Squid hydrolysate 6% 6 weeks
Squid meal 6%

Note: SBM: Solvent extracted soybean meal.

initially calculated assuming a 1.8 feed conversion ratio (FCR) and a
doubling in size the rst week and 0.81.0 g/week thereafter. Daily al-
lowances of feed were adjusted based on observed mortality, weekly
counts of the shrimp and observed feed consumption. The feed was of-
fered in slight excess. Shrimp were fed four times daily and the time in-
terval of feeding is at least 2 h.
In Trial 2, the recirculating system was composed of twenty-eight
square tanks (135 L, 0.52 0.52 0.48 m) water pump, supplemental
aeration unitizing a central line, air diffusers and regenerative blower
as well as mechanical and biological ltration. Four replicate groups of
shrimp (0.80 g initial mean weight, 10 shrimp/tank) were offered
diets over 6 weeks. To produce a more aggressive feeding strategy to
support potentially higher consumption, the typical ration as described
above was increased by 10%. Feed consumption was observed daily, and
when feed was observed to be limiting, the feed inputs were increased.
Shrimp were fed four times daily.
In Trial 3, juvenile shrimp (initial mean weight 1.51 g) were stocked
at a density of 8 shrimp per tank. The recirculating system was com-
posed of twenty circular plastic tanks (160 L, 0.45 m height 0.56 m
upper diameter, 0.80 m lower diameter) water pump, supplemental
aeration unitizing a central line, air diffusers and regenerative blower
as well as mechanical and biological ltration. Shrimp were fed
four times daily for a 6-week period. Based on historic results, feed
inputs were pre-programmed assuming the shrimp would double
their weight weekly up to one gram then gain 0.81.3 g weekly
with a FCR of 1.8. Every tank was provided with two air stones and
covered with netting to prevent losses caused by jumping. Shrimp
were counted weekly and feed inputs were adjusted based on ob-
served survival.
At the end of the growth trials, the shrimp were group weighed and
enumerated. Mean nal weight, biomass, percent weight gain, feed con-
version ratio (FCR), and survival were determined.

Table 3
Composition (g/100 g as is) of plant-based test diet designed to contain 35% protein and 8% lipids in Trial 1.

Basal Squ3 Squ6 Squ9 Sca3 Sca6 Sca9 ISq6 ISc6

Soybean meala 56.80 52.50 48.20 44.00 53.40 50.10 46.80

Corn protein concentrateb 5.80 5.80 5.80 5.80 5.80 5.80 5.80 5.80 5.80
Dry squid hydrolysatec 3.00 6.00 9.00
Dry scallop hydrolysatec 3.00 6.00 9.00
Impregnated SBM with squid hydrolysatec 56.80
Impregnated SBM with scallop hydrolysatec 56.80
Corn starchd 0.14 1.69 3.25 4.70 1.05 1.86 2.67 0.22 1.16
Whole wheatd 25.00 25.00 25.00 25.00 25.00 25.00 25.00 25.00 25.00
Fish oile 6.11 5.86 5.60 5.35 5.60 5.09 4.58 6.03 5.09
Lecithinf 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00
Cholesterolg 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05
Trace mineral premixh 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.50
Dicalcium phosphatei 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50
Vitamin premixj 1.80 1.80 1.80 1.80 1.80 1.80 1.80 1.80 1.80
Choline chlorided 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20
Stay Ck 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10

Proximate composition (as is)

Crude protein 36.14 36.48 36.29 36.38 35.43 35.62 36.25 37.68 37.65
Lipid 7.97 8.60 8.64 9.08 8.48 9.06 8.85 9.19 8.47
Crude ber 3.96 3.82 3.39 2.96 3.24 3.33 3.06 3.34 3.42
Ash 6.48 6.57 6.42 6.31 6.53 6.55 6.55 6.57 7.08
Moisture 7.23 6.36 6.34 6.84 7.37 8.25 7.32 6.53 4.99

Note: Squ: Dry squid hydrolysate; Sca: Dry scallop hydrolysate; ISq: Impregnated soybean meal with squid hydrolysate; ISc: Impregnated soybean meal with scallop hydrolysate.
a
Solvent extracted, Soybean meal Bunge, Decatur, Alabama.
b
Empyreal 75, Cargill Corn Milling, Blair, Nebraska.
c
Produced for this research.
d
MP Biochemicals Inc., Solon, Ohio.
e
Omega Protein Inc., Reedville, Virginia.
f
Solae Company, St. Louis, Missouri.
g
USB Biochemicals, Cleveland, Ohio.
h
Trace mineral premix(g/100 g premix): Cobalt chloride, 0.004; Cupric sulfate pentahydrate, 0.550; Ferrous sulfate, 2.000; Magnesium sulfate anhydrous, 13.862; Manganese sulfate
monohydrate, 0.650; Potassium iodide, 0.067; Sodium selenite, 0.010; Zinc sulfate heptahydrate, 13.193; Alpha-cellulose, 69.664.
i
J. T. Baker, Mallinckrodt Baker, Inc., Phillipsburg, New Jersey.
j
Vitamin premix (g/kg premix): Thiamin.HCL, 4.95; Riboavin, 3.83; Pyridoxine.HCL, 4.00; Ca-Pantothenate, 10.00; Nicotinic acid, 10.00; Biotin, 0.50; folic acid, 4.00; Cyanocobalamin,
0.05; Inositol, 25.00; Vitamin A acetate (500,000 IU/g), 0.32; Vitamin D3 (1,000,000 IU/g), 0.05; Vitamin E Acetate (250 IU/g), 80.00; Menadione, 0.50; Alpha-cellulose, 756.81.
k
Stay C, (L-ascorbyl-2-polyphosphate 25% Active C), DSM Nutritional Products., Parsippany, New Jersey.
56 Y. Zhou et al. / Aquaculture 465 (2016) 5359

Table 4 Table 5
Composition (g/100 g as is) of plant based diets (35% protein and 8% lipid) in Trial 2.a, b, c, Ingredient composition of three experimental diets used in Trial 3 with different style of
d, e, f, g, h, i, j, k attractant in plant-based diet for Litopenaeus vannamei.

Basal-2 Squ32 Sca32 Ingredient (As is) Basal-3 Squ63 Squid63

Soybean meala 54.10 49.80 50.75 Soybean meala 54.10 45.60 44.70
Corn protein concentrateb 8.00 8.00 8.00 Corn protein concentrateb 8.00 8.00 8.00
Dry squid hydrolysatec 3.00 Menhaden sh oilc 6.05 5.55 6.06
Dry scallop hydrolysatec 3.00 Corn starchd 0.70 3.70 4.09
Corn starchd 0.70 2.25 1.56 Whole wheatd 25.00 25.00 25.00
Whole wheatd 25.00 25.00 25.00 Trace mineral premixe 0.50 0.50 0.50
Menhaden sh oile 6.05 5.80 5.54 Vitamin premixf 1.80 1.80 1.80
Lecithinf 1.00 1.00 1.00 Choline chlorided 0.20 0.20 0.20
Cholesterolg 0.05 0.05 0.05 Stay Cg 0.10 0.10 0.10
Trace mineral premixh 0.50 0.50 0.50 Mono-basic calcium phosphateh 2.50 2.50 2.50
Dibasic calcium phosphatei 2.50 2.50 2.50 Soy lecithini 1.00 1.00 1.00
Vitamin premixj 1.80 1.80 1.80 Cholesterolj 0.05 0.05 0.05
Choline chlorided 0.20 0.20 0.20 Dry squid hydrolysatek 6.00
Stay Ck 0.10 0.10 0.10 Squid meall 6.00

Proximate composition (as is) Proximate composition (as is)

Crude protein 37.15 37.98 37.44 Crude protein 37.67 37.14 35.88
Lipid 8.07 9.99 8.05 Lipid 9.54 9.14 8.74
Crude ber 4.35 4.03 3.91 Crude ber 4.05 3.36 3.46
Ash 6.24 6.24 6.34 Ash 6.06 5.68 5.47
Moisture 6.63 5.39 6.43 Moisture 5.41 8.26 10.82

Note: Squ: Dry squid hydrolysate; Sca: Dry scallop hydrolysate. Note: Squ: Dry squid hydrolysate; Squid: Squid meal.
a a
Solvent Extracted, Soybean meal Bunge, Decatur, Alabama. De-hulled solvent extract soybean meal, Bunge Limited, Decatur, Alabama.
b b
Empyreal 75, Cargill Corn Milling, Blair, Nebraska. Empyreal 75, Cargill Corn Milling, Blair, Nebraska.
c c
Produced for this research. Omega Protein Inc., Houston, Texas.
d d
MP Biochemicals Inc., Solon, Ohio. MP Biomedicals Inc., Solon, Ohio.
e e
Omega Protein Inc., Reedville, Virginia. Trace mineral premix (g/100 g premix): Cobalt chloride, 0.004; Cupric sulfate
f
Solae Company, St. Louis, Missouri. pentahydrate, 0.550; Ferrous sulfate, 2.000; Magnesium sulfate anhydrous, 13.862; Man-
g
USB Biochemicals, Cleveland, Ohio. ganese sulfate monohydrate, 0.650; Potassium iodide, 0.067; Sodium selenite, 0.010; Zinc
h
Trace mineral premix (g/100 g premix): Cobalt chloride, 0.004; Cupric sulfate sulfate heptahydrate, 13.193; Alpha-cellulose, 69.664.
f
pentahydrate, 0.550; Ferrous sulfate, 2.000; Magnesium sulfate anhydrous, 13.862; Man- Vitamin premix (g/kg premix): Thiamin.HCL, 4.95; Riboavin, 3.83; Pyridoxine.HCL,
ganese sulfate monohydrate, 0.650; Potassium iodide, 0.067; Sodium selenite, 0.010; Zinc 4.00; Ca-Pantothenate, 10.00; Nicotinic acid, 10.00; Biotin, 0.50; folic acid, 4.00; Cyanoco-
sulfate heptahydrate, 13.193; Alpha-cellulose, 69.664. balamin, 0.05; Inositol, 25.00; Vitamin A acetate (500,000 IU/g), 0.32; Vitamin D3
i
J. T. Baker, Mallinckrodt Baker, Inc., Phillipsburg, New Jersey. (1,000,000 IU/g), 80.00; Menadione, 0.50; Alpha-cellulose, 856.81.
j g
Vitamin premix (g/kg premix): Thiamin.HCL, 4.95; Riboavin, 3.83; Pyridoxine.HCL, Stay C, (L-ascorbyl-2-polyphosphate 35% Active C), DSM Nutritional Products.,
4.00; Ca-Pantothenate, 10.00; Nicotinic acid, 10.00; Biotin, 0.50; folic acid, 4.00; Cyanoco- Parsippany, New Jersey.
h
balamin, 0.05; Inositol, 25.00; Vitamin A acetate (500,000 IU/g), 0.32; Vitamin D3 Calcium dihydrogen phosphate dydrate - alpha aesar.
i
(1,000,000 IU/g), 0.05; Vitamin E Acetate (250 IU/g), 80.00; Menadione, 0.50; Alpha-cellu- The Solae Company, St. Louis, Missouri.
j
lose, 756.81. USB Biochemicals, Cleveland, Ohio.
k k
Stay C, (L-ascorbyl-2-polyphosphate 25% Active C), DSM Nutritional Products., Produced by University of Rhode Island, Kingston, Rhode Island.
l
Parsippany, New Jersey. Foodcorp S.A., Chile.

2.4. Feed intake
To evaluate possible increases in consumption, the test diets in Trial
2 were used in a consumption trial. Shrimp were pre-conditioned to the
test diets then four replicate groups of 10 shrimp (2.99 g mean weight)
were stocked into the same recirculating system as in Trial 2. Consump-
tion was determined by offering an excess of feed for three 1 h feedings
each day over a three-day period. After one hour of access to the feed,
uneaten feed was collected by siphoning onto a 500 m mesh screen,
quantied and feed intake estimated on a dry weight basis.
2.5. Water quality monitoring
In the three trials, temperature, dissolved oxygen (DO), pH, and sa-
linity were monitored twice daily using YSI 556 m (Yellow Spring In-
strument Co., Yellow Spring, Ohio). Total ammonia nitrogen and
nitrite were measured weekly described by Solorzano (1969) and
Spotte (1979), respectively.
2.6. Statistical analysis
Data were analyzed using a one-way analysis of variance to deter-
mine if signicant P b 0.05 differences existed among treatment
means in each trial. The StudentNeumanKeuls multiple comparison
test was used to determine where signicant differences existed
among treatment means in each trial (Steel and Torrie, 1980). The
pooled standard errors were used across all the trials, as the variance
of each treatment is the same. All statistical analyses were conducted
using the SAS 9.4 (SAS Institute, Cary, North Carolina).
3. Results
3.1. Water quality
In Trial 1, water temperature (mean SD) were recorded at
30.12 0.56 C, pH at 7.23 0.14, salinity at 5.18 0.22, and DO
at 6.44 0.17 mg/L. Water quality parameters for Trial 2 were as fol-
lows: temperature at 26.47 2.52 C, pH at 7.40 0.28, salinity at
5.36 0.25, DO at 6.15 0.49 mg/L, total ammonia nitrogen at
0.17 0.05 mg/L, and nitrite at 0.16 0.07 mg/L. Water quality param-
eters for Trial 3 were as follows: temperature at 28.14 1.64 C, pH at
7.75 0.20, salinity at 9.58 0.67, DO at 5.98 0.41 mg/L, total am-
monia nitrogen at 0.08 0.09 mg/L, and nitrite at 0.03 0.02 mg/L.
Water quality conditions in the trials were suitable for the normal
growth and survival of this species.
3.2. Growth performance
In Trial 1, nal biomass ranged from 55.02 to 64.99 g, nal mean
weight of the shrimp was between 6.49 and 7.27 g, percent weight
gain varied from 896.0 to 1027.0%, FCR was between 1.41 and 1.60,
 Y. Zhou et al. / Aquaculture 465 (2016) 5359 57

and survival ranged from 80.0 to 95.0% (Table 6). In Trial 1, there were Table 7
no signicant differences with regard to any treatment on the shrimp's In Trial 2, response of juvenile shrimp (0.8 g mean initial weight) to three soybean meal
based offered over a 6-week period. Values are means of four replicates. Estimated feed in-
nal biomass, nal mean weight, percent weight gain, FCR, and survival. take was determined using a second group of shrimp (3 g mean weight) over a three-day
In Trial 2, nal biomass ranged from 74.85 to 83.85 g, nal mean period. As there was no signicant effect of day, the presented data is averaged across all
weight of the shrimp was between 8.09 and 8.58 g, percent weight three days n = 12 (4 replicate tanks for 3 days). Means within columns with the same let-
gain varied from 922.69 to 1006.2%, FCR was between 1.45 and 1.59, ter are not signicant different (P N 0.05) based on analysis of variance followed by Stu-
dent Newman-Keuls multiple range test.
and survival ranged from 90.0 to 97.5% (Table 6). In Trial 2, no signi-
cant differences were found on the shrimp's nal biomass, nal mean Final
weight, percent weight gain, FCR, and survival. In the feed consumption biomass Final mean Weight Survival Estimated
Diet (g) weight (g) gain (%) FCRa1 (%) intake (g)
trial, the value of estimated intake in basal diet was signicantly higher
than the values of other diets (Table 7). Basal-2 74.85 8.09 922.69 1.59 92.5 16.35a
In Trial 3, nal mean weight of the shrimp was between 8.26 and Squ32 79.20 8.83 1006.08 1.45 90.0 14.31b
Sca32 83.58 8.58 980.93 1.50 97.5 13.53b
9.67 g, percent weight gain varied from 440.04 to 537.37%, FCR was be- PSE2 2.92 0.21 30.91 0.04 3.91 0.56
tween 1.34 and 1.65, and survival ranged from 75.00 to 87.50% P-value 0.1628 0.0821 0.2029 0.056 0.4208 0.0003
(Table 8). Analysis of variance indicated a signicant effect on nal bio-
Note: Squ: dry squid hydrolysate; Sca: dry scallop hydrolysate.
mass (P = 0.0498) with a ranged from 49.25 to 67.53 g, However, mean 1
FCR: Feed conversion ratio.
separation could not detect differences. In Trial 3, no signicant differ- 2
PSE: Pool standard error.
ences were found on the shrimp's nal biomass, nal mean weight, per-
cent weight gain, FCR, and survival.
found as feeding stimulants for Pacic white shrimp over a 4-day peri-
4. Discussion od. However, so far there is limited information for the squid/scallop hy-
drolysate in the diets for testing the growth performance of Pacic
As the level of plant-based feedstuffs increases in feed formulations, white shrimp.
there is a perception of reduced consumption and consequently growth. Feeding attractants have been generally supplemented ranging from
Hence, it has been recommended that research into the area of attrac- 0.5% to 5.0% in diet of shrimp dependent on the attractant and diet for-
tants and palatability enhancers or supplements to enhance growth mulations (Derby et al., 2016; Hartati and Briggs, 1993). In the present
performance in plant-based diets should receive more attention in fu- study, no signicant differences with regard to growth performance
ture years (Browdy et al., 2007; Gatlin and Li, 2008). Marine feedstuffs are observed among any treatment in Trial 1 (Table 6), when the diets
including sh meal, sh oil, squid meal, krill meal, sh soluble, and var- were supplemented at 3%, 6%, and 9% Squ or Sca. Results of Trial 2
ious protein hydrolysates are well known to be highly palatable to var- (Table 7) also conrmed that there are no signicant differences regard-
ious aquatic animals (Barrows and Hardy, 2000; Carr and Derby, 1986; ing growth performance with the similar basal diet supplemented with
Derby et al., 2016; Harpaz, 1997; Holland and Borski, 1993; Sanchez 3% Squ or Sca, it should be noted that hydrolysate-added diets brought
et al., 2005; Smith et al., 2005; Suresh et al., 2011). In sh, several previ- about some increases in nal mean weight and weight gain, and lower
ous studies have demonstrated the protein hydrolysates from by- FCR, especially squid hydrolysate. Likewise, results of Trial 3 (Table 8)
products of marine sources would provide the benecial effects in Gold- conrmed that there are no signicant differences regarding growth
sh Carassius auratus L. (Szlaminska et al., 1991), Common Carp performance with the similar basal diet supplemented with 6% Squ or
Cyprinus carpio L. (Carvalho et al., 1997), Sea Bass Dicentrarchus labrax squid meal. Protein hydrolysates as an attractant in crustacean have
(Infante et al., 1997). Lian et al. (2005) and Lian et al. (2008) reported been assessed with ambiguous or marginal results: improvements in
that squid hydrolysate can be a good source of aquaculture feed ingredi- growth in Pacic white shrimp (Anggawati et al., 1990;
ent, particularly for the diet of larval sh. In blue shrimp Litopenaeus Crdova-Murueta and Garca-Carreo, 2002; Floreto et al., 2001;
stylirostris, Suresh et al. (2011) reported that 3% sh hydrolysate can im- Hernndez et al., 2011) or giant tiger prawn P. monodon, (Anggawati
prove the attractability of the feeds. Grey et al. (2009) documented that et al., 1990), while no signicant effect of growth were observed in
four different kinds of salmon hydrolysate tested (liquid acidied, acid- giant tiger prawn (Hartati and Briggs, 1993; Smith et al., 2005). This
ied dried, nonacidied dried, neutralized dried, included at 5%) were phenomenon is attributed to many factors such as the supplemental
level, variation of quality for protein hydrolysates (different amino
acids prole etc.), the different diet formulation (re-pelleted commer-
Table 6
In Trial 1, response of juvenile shrimp (Mean initial weight 0.66 0.03 g) offered plant
cial feed, practical diet, or semi-puried diets), and different rearing pe-
based test diets with squid or scallop hydrolysate at varying levels over 6 weeks. Values riod and conditions as well as different species. For example, Crdova-
represent the mean of four replicates. Means within columns with the same letter are Murueta and Garca-Carreo (2002) reported that they used a re-
not signicant different (P N 0.05) based on analysis of variance followed by Student New- pelleted commercial feed supplemented with either krill hydrolysate
man-Keuls multiple range test.
or sh hydrolysate, and indicated the nal weight of shrimp with 3%,
Final biomass Final mean weight Weight gain Survival
Diet (g) (g) (%) FCRa (%) Table 8
Basal 59.50 7.21 990.4 1.45 82.5 In Trial 3, effect of different attractant on growth performance of Litopenaeus vannamei
Squ3 55.07 6.49 896.0 1.60 85.0 over 6 weeks. Values are means of four replicates. Means within columns with the same
Squ6 64.55 6.81 923.8 1.50 95.0 letter are not signicant different (P N 0.05) based on analysis of variance followed by Stu-
Squ9 59.32 7.25 1004.3 1.42 82.5 dent Newman-Keuls multiple range test.
Sca3 55.02 6.70 936.7 1.54 82.5
Final mean Final biomass Weight gain Survival
Sca6 58.17 7.06 958.8 1.46 82.5
Trt weight (g) (g) (%) FCRa (%)
Sca9 63.29 7.04 956.4 1.45 90.0
ISq6 64.99 7.27 1027.0 1.41 90.0 Basal-3 8.26 49.25z 440.04 1.65 75.00
ISc6 57.24 7.18 981.9 1.43 80.0 Squ63 9.67 67.53z 537.37 1.34 87.50
PSEb 3.12 0.22 42.05 0.05 5.35 Squid63 8.40 53.18z 465.03 1.60 78.13
P-value 0.2047 0.1729 0.4812 0.1565 0.5617 PSEb 0.4027 4.6585 35.2957 0.1085 5.4127
P-value 0.0677 0.0498 0.1846 0.1476 0.2857
Note: Squ: Dry squid hydrolysate; Sca: Dry scallop hydrolysate, ISq: Impregnated soybean
meal with squid hydrolysate; ISc: Impregnated soybean meal with scallop hydrolysate. Note: Squ: dry squid hydrolysate; Squid: squid meal.
a a
FCR = Feed conversion ratio. FCR: Feed conversion ratio.
b b
PSE = Pooled standard error. PSE: Pool standard error.
58 Y. Zhou et al. / Aquaculture 465 (2016) 5359
9%, 15% krill hydrolysate or 3%, 9% sh hydrolysate were signicantly
higher than the control feed. This may be caused by inadequacies of
the diet formulation of the commercial feed with lower protein or
other reasons, as no detailed composition information of the commer-
cial feed is reported. Another factor for the inconsistent results with
supplemental protein hydrolysates is different styles of protein hydroly-
sates with different processing procedure in these studies such as the
tuna by-product protein hydrolysates (Hernndez et al., 2011), krill hy-
drolysate (Crdova-Murueta and Garca-Carreo, 2002; Floreto et al.,
2001; Smith et al., 2005), and sh hydrolysates (Anggawati et al.,
1990; Crdova-Murueta and Garca-Carreo, 2002; Smith et al., 2005).
Thus, it is not surprising that protein hydrolysate supplementation
with ambiguous results are affected by many factors. In a series of stud-
ies conducted with different sh species (Lee et al., 2015), all species
responded favorably to squid and scallop hydrolysates including juve-
nile summer ounder Paralichthys dentatus, Atlantic salmon Salmo
salar and yellowtail favors squid hydrolysate, while European seabass
Dicentrarchus labrax performed better with scallop hydrolysate suggest-
ing species dependency. These results suggest that the Pacic white
shrimp may not respond to hydrolysate as well as sh. Overall, the pro-
tein hydrolysates could be supplemented as a nutrient source in the diet
of shrimp should it be cost effective.
Our ndings indicate that supplementing Squ or Sca to the basal diet
did not clearly improve the performance of shrimp in terms of nal
mean weight, or percent weight gain (Table 6). Hernndez et al.
(2011) observed that the nal body weight and specic growth rate of
shrimp with inclusion of 2.5% and 5.0% tuna by-product protein hydro-
lysates (TBPH) were signicantly increased. They concluded that free
amino acids in protein hydrolysates can improve the absorption of pro-
tein. However, Hernndez et al. (2011) also found that the nal body
weight and specic growth rate of shrimp with 7.5% and 10.0% TBPH
signicantly decreased compared to shrimp fed feeds with 2.5% and
5.0% TBPH. However, in the current study, the nal mean weight and
percent weight gain did not show signicant differences.
In general, the high levels of plant protein in diets of shrimp could
negatively inuence the dietary amino acid composition, and palatabil-
ity, which may reduce the feed intake and performance of shrimp
(Bulbul et al., 2015; Bulbul et al., 2013; Lim and Dominy, 1990;
Samocha et al., 2004). To evaluate possible shifts in consumption, feed
intake was measured in a separate group of shrimp. The estimated in-
take of shrimp offered the basal diet was signicantly higher than that
of shrimp offered diets supplemented with 3% Squ or Sca (Table 7). It
is generally viewed that squid meal and other marine by-products are
good attractants in shrimp diets which increase consumption, studies
are not always positive and there are numerous interactions which
could inuence the results. Furthermore, intake does not always follow
growth rate. For example, Smith et al. (2005) reported that (2% inclu-
sion) sh hydrolysate and (1%) krill hydrolysate meal did not improve
the growth rate, FCR, survival, and daily apparent feed intake of giant
tiger prawn. Sanchez et al. (2012) demonstrated that the Pacic white
shrimp reared in clear water or green water systems fed the diets con-
taining sh meal (6.5 or 12.0%) and squid meal (5%, 10%, or 20%) did
not show any signicant differences in FCR or weight gain. In any case,
under the reported conditions there was no benet or disadvantage in
consumption of feed when the hydrolysates were added to the diets.
Under the reported conditions of the study, the use of Squ and Sca
did not signicantly improve growth or feed intake of Pacic white
shrimp offered a plant-based diet. Although there were no improve-
ments there were also no depressions in performance. Hence, the data
in the present study demonstrates that these products created from
waste streams of squid and scallop processing centers are good nutrient
sources and can be incorporated in practical shrimp feed formulations if
it proves to be cost effective. Under our conditions, they did not seem to
function as attractants, since they did not improve growth or feed intake.
The inclusion of these marine by-products in plant-based diets for shrimp
should be further investigated as they contain essential nutrients.
Acknowledgments
The authors would like to thank you those who have taken the time
to critically review this manuscript. This work was partially supported
by the NOAA Fisheries Service and HATCH funding, in cooperation
with the Saltonstall-Kennedy Grant Program. The mention of trade-
marks or proprietary products does not constitute an endorsement of
the product by Auburn University and does not imply its approval to
the exclusion of other products that may also be suitable.
References
Akiyama, T., Murai, T., Hirasawa, Y., Nose, T., 1984. Supplementation of various meals to sh-
meal diet for chum salmon fry. Aquaculture 37, 217222.
Aksnes, A., Hope, B., Hostmark, O., Albrektsen, S., 2006. Inclusion of size fractionated sh hydro-
lysate in high plant protein diets for Atlantic cod, Gadus morhua. Aquaculture 261,
11021110.
Anggawati, A., Murtini, J., Heruwati, E., 1990. The Use of Hydrolyzed Protein Concentrate in Prac-
tical Diets for Penaeus monodon Juveniles. Research Report. Research Institute for Fish Tech-
nology, Palmerah Jakarta, Indonesia, p. 12.
Barrows, F.T., Hardy, R.W., 2000. Feed Additives. In: Stickney, R.R. (Ed.), Enclyclopedia of Aqua-
culture. Wiley, New York, pp. 335340.
Browdy, C., Seaborn, G., Atwood, H., Davis, D.A., Bullis, R.A., Samocha, T.M., Wirth, E., Lefer, J.W.,
2007. Comparison of pond production efciency, fatty acid proles, and contaminants in
Litopenaeus vannamei fed organic plant-based and fish-meal-based diets. J. World Aquacult.
Soc. 37, 437451.
Bui, H.T.D., Khosravi, S., Fournier, V., Herault, M., Lee, K.-J., 2014. Growth performance, feed uti-
lization, innate immunity, digestibility and disease resistance of juvenile red seabream
(Pagrus major) fed diets supplemented with protein hydrolysates. Aquaculture 418, 1116.
Bulbul, M., Koshio, S., Ishikawa, M., Yokoyama, S., Kader, M.A., 2013. Performance of kuruma
shrimp, Marsupenaeus japonicus fed diets replacing shmeal with a combination of plant
protein meals. Aquaculture 372-375, 4551.
Bulbul, M., Koshio, S., Ishikawa, M., Yokoyama, S., Abdul Kader, M., 2015. Growth performance of
juvenile kuruma shrimp, Marsupenaeus japonicus (bate) fed diets replacing shmeal with
soybean meal. Aquac. Res. 46, 572580.
Carr, W.E.S., Derby, C.D., 1986. Chemically stimulated feeding behavior in marine animals.
J. Chem. Ecol. 12, 9891011.
Carvalho, A.P., Escaffre, A.M., Oliva Teles, A., Bergot, P., 1997. First feeding of common carp larvae
on diets with high levels of protein hydrolysates. Aquac. Int. 5, 361367.
Chotikachinda, R., Tantikitti, C., Benjakul, S., Rustad, T., Kumarnsit, E., 2013. Production of protein
hydrolysates from skipjack tuna (Katsuwonus pelamis) viscera as feeding attractants for
Asian seabass (Lates calcarifer). Aquac. Nutr. 19, 773784.
Crdova-Murueta, J.H., Garca-Carreo, F.L., 2002. Nutritive value of squid and hydrolyzed pro-
tein supplement in shrimp feed. Aquaculture 210, 371384.
Deng, D.F., Ju, Z.Y., Dominy, W.G., Bechtel, P.J., Smiley, S., 2013. An evaluation of pink salmon
(Oncorhynchus gorbuscha) testes meal in diets for pacic white shrimp (Litopenaeus
vannamei): effect on palatability, digestibility and growth performance. Aquac. Nutr. 19,
908916.
Derby, C.D., Elsayed, F.H., Williams, S.A., Gonzlez, C., Choe, M.N., Bharadwaj, A.S., Chamberlain,
G.W., 2016. Krill meal enhances performance of feed pellets through concentration-
dependent prolongation of consumption by Pacic white shrimp, Litopenaeus vannamei.
Aquaculture 458, 1320.
Fetter, J.C., Brown, R.N., Grres, J.H., Chong, L., Amador, J.A., 2012. Nitrate and phosphate leaching
under Turfgrass fertilized with a squid-based organic fertilizer. Water Air Soil Pollut. 223,
15311541.
Floreto, E., Brown, P., Bayer, R., 2001. The effects of krill hydrolysate-supplemented soya-bean
based diets on the growth, colouration, amino and fatty acid proles of juvenile
American lobster, Homarus americanus. Aquac. Nutr. 7, 3343.
Gatlin III, D., Li, P., 2008. Use of Diet Additives to Improve Nutritional Value of Alternative Protein
Sources. In: Lim, C., Webster, C.D., Lee, C.-S. (Eds.), Alternative Protein Sources in Aquacul-
ture Diets. The Haworth Press, New York, USA, pp. 501522.
Grey, M., Forster, I., Dominy, W., Ako, H., Giesen, A.F., 2009. Validation of a feeding stimulant bio-
assay using sh hydrolysates for the Pacic white shrimp, Litopenaeus vannamei. J. World
Aquacult. Soc. 40, 547555.
Harpaz, S., 1997. Enhancement of growth in juvenile freshwater prawn Macrobrachium
rosenbergii through the use of a chemoattractant. Aquaculture 156, 221227.
Hartati, R., Briggs, M., 1993. Effect of feeding attractants on the behaviour and performance of
juvenile Penaeus monodon Fabricius. Aquac. Res. 24, 613624.
Hernndez, C., Olvera-Novoa, M.A., Smith, D.M., Hardy, R.W., Gonzalez-Rodriguez, B., 2011. En-
hancement of shrimp Litopenaeus vannamei diets based on terrestrial protein sources via
the inclusion of tuna by-product protein hydrolysates. Aquaculture 317, 117123.
Holland, K.N., Borski, R.J., 1993. A palatability bioassay for determining ingestive stimuli in the
marine shrimp Penaeus wannamei. Aquaculture 109, 153164.
Infante, J.L.Z., Cahu, C.L., Peres, A., 1997. Partial substitution of di-and tripeptides for native pro-
teins in sea bass diet improves Dicentrarchus labrax larval development. J. Nutr. 127,
608614.
Kader, M.A., Bulbul, M., Koshio, S., Ishikawa, M., Yokoyama, S., Nguyen, B.T., Komilus, C.F.,
2012. Effect of complete replacement of shmeal by dehulled soybean meal with
crude attractants supplementation in diets for red sea bream, Pagrus major. Aquacul-
ture 350, 109116.
Karayannakidis, P.D., Apostolidis, E., Lee, C.M., 2014. Comparison of direct steam injection and
steam-jacketed heating in squid protein hydrolysis for energy consumption and hydrolysis
performance. LWT Food Sci. Technol. 57, 134140.
Lee, C.M., 2004. Bioconversion of Squid Processing Waste for the Production of Specialty Aqua-
culture Feed Ingredients. Technical Report to NOAA-NMFS (S-K Program NA16FD2299, 01-
NER-010).
 Y. Zhou et al. / Aquaculture 465 (2016) 5359
Lee, C.M., Lian, P., 2010. Bioproduction of hydrolysate from squid processing byproducts for
aquaculture feed ingredient and organic fertilizer. US Patent Application No: 2010/
0175,441.
Lee, C.M., Volson, B., Kang, B., Karayannakidis, P.D., Gamez, E., Miller, J., Bengtson, D.A., Wolters,
W.R., Burr, G.S., Barrows, R., 2015. Hydrolysates from Scallop and Squid Processing
Byproducts as Specialty Aquafeed Ingredients, the Annual International Conference of
World Aquaculture Society Conference. May 2630, Jeju, Korea.
Lian, P.Z., Lee, C.M., 2003a. Characterization of Hydrolysate from Squid Processing Waste and its
Feed Attractability to Trout Fingerling, Proceedings of the 2003 IFT Annunal Meeting. July
1216, 2003, Chicago, Illinois, USA..
Lian, P.Z., Lee, C.M., 2003b. Characterization of Squid Hydrolysate for its Potential as Aquaculture
Feed Ingredient, Proceeding of the First Joint Trans-Atlantic Fisheries Technology Confer-
ence (TAFT). June 1014, 2003, Reykjavik, Iceland.
Lian, P., Lee, C., Park, E., 2005. Characterization of squid-processing byproduct hydrolysate and
its potential as aquaculture feed ingredient. J. Agric. Food Chem. 53, 55875592.
Lian, P., Lee, C.M., Bengtson, D.A., 2008. Development of a squid-hydrolysate-based larval diet
and its feeding performance on summer ounder, Paralichthys dentatus, larvae. J. World
Aquacult. Soc. 39, 196204.
Lim, C., Dominy, W., 1990. Evaluation of soybean meal as a replacement for marine animal pro-
tein in diets for shrimp (Penaeus vannamei). Aquaculture 87, 5363.
Murray, A.L., Pascho, R.J., Alcorn, S.W., Fairgrieve, W., Shearer, K., Roley, D., 2003. Effects of var-
ious feed supplements containing sh protein hydrolysate or sh processing by-products
on the innate immune functions of juvenile coho salmon (Oncorhynchus kisutch). Aquacul-
ture 220, 643653.
Nagel, F., von Danwitz, A., Schlachter, M., Kroeckel, S., Wagner, C., Schulz, C., 2014. Blue mussel
meal as feed attractant in rapeseed protein-based diets for turbot (Psetta maxima L.). Aquac.
Res. 45, 19641978.
Nguyen, H.T.M., Prez-Glvez, R., Berg, J.P., 2012. Effect of diets containing tuna head hydroly-
sates on the survival and growth of shrimp Penaeus vannamei. Aquaculture 324, 127134.
Nunes, A.J., S, M.V., Andriola-Neto, F.F., Lemos, D., 2006. Behavioral response to selected feed
attractants and stimulants in Pacic white shrimp, Litopenaeus vannamei. Aquaculture
260, 244254.
Oliva-Teles, A., Cerqueira, A.L., Goncalves, P., 1999. The utilization of diets containing high levels
of sh protein hydrolysate by turbot (Scophthalmus maximus) juveniles. Aquaculture 179,
195201.
Refstie, S., Olli, J.J., Standal, H., 2004. Feed intake, growth, and protein utilisation by post-smolt
Atlantic salmon (Salmo salar) in response to graded levels of sh protein hydrolysate in
the diet. Aquaculture 239, 331349.
59
Samocha, T.M., Davis, D.A., Saoud, I.P., DeBault, K., 2004. Substitution of sh meal by co-extruded
soybean poultry by-product meal in practical diets for the Pacic white shrimp, Litopenaeus
vannamei. Aquaculture 231, 197203.
Sanchez, D.R., Fox, J.M., Lawrence, A.L., Castille, F.L., Dunsford, B., 2005. A methodology for eval-
uation of dietary feeding stimulants for the Pacic white shrimp, Litopenaeus vannamei.
J. World Aquacult. Soc. 36, 1423.
Sanchez, D.R., Fox, J.M., Gatlin, D., Lawrence, A.L., 2012. Dietary effect of squid and sh meals on
growth and survival of Pacic white shrimp Litopenaeus vannamei in the presence or ab-
sence of phytoplankton in an indoor tank system. Aquac. Res. 43, 18801890.
Smith, D.M., Tabrett, S.J., Barclay, M.C., Irvin, S.J., 2005. The efcacy of ingredients included in
shrimp feeds to stimulate intake. Aquac. Nutr. 11, 263272.
Solorzano, L., 1969. Determination of ammonia in natural waters by the phenolhypochlorite
method. Limnol. Oceanogr. 14, 799801.
Spotte, S., 1979. Fish and Invertebrate Culture: Water Management in Closed Systems. 2nd ed.
Wiley, New York.
Steel, R.G.D., Torrie, J.H., 1980. Principles and Procedures of Statistics: A Biometrical Approach.
McGraw-Hill, New York, USA.
Suresh, A.V., Vasagam, K.P.K., Nates, S., 2011. Attractability and palatability of protein ingredi-
ents of aquatic and terrestrial animal origin, and their practical value for blue shrimp,
Litopenaeus stylirostris fed diets formulated with high levels of poultry byproduct meal.
Aquaculture 319, 132140.
Szlaminska, M., Escaffre, A., Charlon, N., Bergot, P., 1991. Preliminary Data on Semisynthetic
Diets for Goldsh (Carassius auratus L.) Larvae. In: Kaushik, S.J., Luquet, P. (Eds.), Fish Nutri-
tion in Practice. INRA, Paris, Les Colloques Biarritz, France, pp. 607612.
Watanabe, T., 2002. Strategies for further development of aquatic feeds. Fish. Sci. 68, 242252.
Watanabe, T., Aoki, H., Shimamoto, K., Hadzuma, M., Maita, M., Yamagata, Y., Kiron, V., Satoh, S.,
1998. A trial to culture yellowtail with non-shmeal diets. Fish. Sci. 64, 505512.
Zheng, K., Liang, M., Yao, H., Wang, J., Chang, Q., 2012. Effect of dietary sh protein hydrolysate
on growth, feed utilization and IGF-I levels of Japanese ounder (Paralichthys olivaceus).
Aquac. Nutr. 18, 297303.
Zhou, Y., Davis, D.A., Rhodes, M.A., 2014. Comparative evaluation of copper sulfate and tribasic
copper chloride on growth performance and tissue response in Pacic white shrimp
Litopenaeus vannamei fed practical diets. Aquaculture 434, 411417.