Download New Advances in Surimi Technology

Squid Hydrolysate for
Larval Fish Feed Ingredients
Chong M. Lee 1, Peizhi Lian 1, David Bengtson 2,
Nick King 3 and George Nardi 4
1. Dept. of Nutrition and Food Sciences
2. Dept. of Fisheries, Animal and Veterinary Science
University of Rhode Island
3. Skretting, Vancouver, BC
4. GreatBay Aquaculture, Portsmouth, New Hampshire
Outline of presentation

Background

Squid hydrolysate production

Feed formulation and production

Field feeding trials

Ongoing and future studies
Background



36,000 MT of squid (Loligo pealei and Illex illecebrosus)
landed in Northeast of U.S. annually
40-50% turns into processing byproduct or waste
currently not being utilized
Presently, the Northeast of U.S. (RI, NY and NJ)
generates ~10 million lbs (4,540 MT) squid
processing byproduct/yr.
Squid byproduct characteristics

Consists of head, fin and viscera
+ unclaimed mantles and tentacles

Approximately, 11% protein,
2% lipid (11.6% EPA; 24.5% DHA),
1.2% ash and 86% moisture

Unique features
- Use of endogenous enzymes for hydrolysis
- Believed to possess a protein fraction associated
with “growth factor” (Meyers, 1989), and high level of
chemo-attractant betaine and gowth promoting taurine
- Squid hydrolysate may offer a potential
as a specialty feed ingredient e.g. starter diet
Commercial scale production of squid hydrolysate
Raw squid byproduct
In-line viscometer
86% M
Hydrolysis
330 gal capacity
Vibrating screen
Evaporator
hydrolysate
73-78% M
Yield: 47-54%
Proximate composition of squid hydrolysate
(concentrated – 76% moisture)
Moisture
Protein
Lipid
Ash
76 %
18.63 %
3.30 %
2.19 %
•Squid hydrolysate contains 11.16% EPA and 24.45% DHA, while
salmon oil contains 8.65% EPA and 10.67% DHA (on an oil weight basis).
Amino acid profiles of squid hydrolysate (based on 76% moisture-containing hydrolysate)
Amino acids
Hydrolysate
weight (%)
Per protein wt
(%)
Asp
1.87
9.46
Free amino acids/
hydrolysate
(mg/g)
6.95
Glu
2.50
12.63
11.14
Ser
0.79
4.01
2.34
Gly
1.06
4.01
4.57
His
0.29
1.45
1.10
Arg
1.98
10.02
14.21
Thr
0.59
2.97
3.97
Ala
0.98
4.96
4.35
Pro
0.85
4.29
5.22
Tyr
0.61
3.11
2.30
Val
0.81
4.08
2.68
Met
0.65
3.32
1.78
Cys
0.10
0.49
0.18
Ile
0.76
3.87
2.23
Leu
1.48
7.47
4.41
Phe
0.74
3.73
3.25
Lys
1.20
6.07
3.54
 Aslo it contains high level of chemo-attractant betaine (Takaoka et al. 1995. Fisheries Science 61: 833-836;
and taurine (Cho SY et al. 2000. J Korean Fish Soc 33: 51-54)
Composition of squid hydrolysate-based basal microdiet
Ingredients
Amount (%
dry wt basis)
Squid hydrolysate
73.33
Salmon oil
9.54
Lecithin
3.01
Vitamin- premix
0.44
Mineral premix
2.01
Starch
5.02
Yeast
4.02
Algae (spirulina: chlorella)
2.64
Proximate composition
Protein
64.66
Lipid
18.72
Carbohydrate
7.40
Ash
9.21
Energy (MJ/Kg)
19.12
Pilot plant production of microparticulate feed
homogenizng
sieving
Grinding
sieving
drum drying
18:2n6c,t
11.00
0.82
10.84
0.61
18:3n6
0.11
0.01
0.11
0.01
18:3n3
1.15
0.07
1.12
0.05
20
0.01
0.02
0.01
0.01
20:1
4.47
0.36
4.46
0.24
Fatty
acid
profiles
of
microdiets
20:2
0.00
0.00
0.00
0.00
20:3n6
0.00150-300 (mm)
0.00
0.00
300-500(mm) 0.00
Fatty
acids
21:0
5.00
0.00
5.00
0.00
AVE
SD
AVE
SD
20:3n3
0.70
0.04
0.70
0.04
10
0.00
0.00
0.00
0.00
20:4n6
0.12
0.01
0.12
0.00
11
0.00
0.00
0.00
0.00
(EPA)
12
0.00
0.00
0.00
0.00
20:5n3
6.34
0.40
6.33
0.43
0.00
0.00
0.00
0.00
2213
0.07
0.01
0.07
0.00
14
1.84
0.22
1.87
0.26
22:1n9
1.99
2.36
0.32
0.02
14:1
0.00
0.00
0.02
0.02
22:2
0.00
0.00
0.00
0.00
0.19
0.02
0.19
0.01
2315
0.00
0.00
0.00
0.00
15:1
0.00
0.00
0.00
0.00
24
0.00
0.00
0.00
0.00
16
11.72
0.84
11.62
0.74
22:6n3 (DHA)
12.31
0.80
12.33
0.77
16:1
2.54
0.23
2.52
0.25
24:1
0.00
0.00
0.00
0.00
17
0.28
0.03
0.27
0.06
1.20
1.21
18.65
18.66
EPA
17:1 + DHA (%)
0.06
0.00
0.06
0.00
Identified comp.
63.27
63.62
18
2.11
0.12
2.08
0.10
Unidentified comp.
36.73
36.38
18:1n9c,t
8.73
0.62
8.60
0.46
18:2n6c,t
11.00
0.82
10.84
0.61
*The
100 g standard
basal squid
hydrolysate
diet0.11
will
2.000.01
g50EPA
andfor
3.60 0.11
g DHA
18:3n6
0.01
*Internal
: C21
(Heneicosanoic
acid provide
methyl ester),
ug/mL
18:3n3
1.15acid/oil).
0.07
1.12
0.05
based
on EPA/DHA
(% fatty
analysis.Fatty
acid distribution
standard: FAMQ-5
FAME
Reference Standard
from
20
0.02
0.01
0.01
AccuStandard,
Inc. New Haven, CT. 0.01
20:1
4.47
0.36
4.46
0.24
20:2
0.00
0.00
0.00
0.00
20:3n6
0.00
0.00
0.00
0.00
21:0
5.00
0.00
5.00
0.00
Field feeding trial on Atlantic cod larvae:
weaning procedure




0.25 million of cod larvae/ production tank (5 m3 ).
One tank for squid hydrolysate(SH)-larval diet
Six tanks (control group) for a premium commercial diet
Cod larvae upon hatch were on
- initially rotifer
- the combination of rotifer and Artemia
- co-feeding of Artemia and microdiet
Weaning 1 week later with gradual removal of Artemia,
the fish were kept on the SH microdiet for another 2
weeks.
(The trial was conducted at the GreatBay Aquaculture in Portsmouth, NH)
Survival


70-75% of the fish on the SH microdiet survived
through the weaning period, which is considered
excellent (in the same range for the control group
- the premium commercial diet)
Overall, there was no real difference in survival
among the production tanks during weaning.
Swimming behavior



Different behavior between the fish fed SH diet
and the rest.
The SH fish had a lighter color. A darker color
is often associated with stress.
The SH fish were very responsive as a sign of
good health being more uniform in size.
This has very significant ramifications as it relates to
cannibalism and grading.

The fish were swimming together in uniform
manner and appeared to be in motion more so
than those in the control diet tanks.
Tank hygiene


The SH diet was rated better than the
premium control diet.
The SH diet appeared to stay very
stable in the water without leaching.
Leaching tends to cause foam on the
surface (which was a problem with the
control diet).
Salinity stress test for cod larvae (48 dph)
42-45
DPH
15.9
+/0.37
16.3
+/0.54
18.3
+/0.41
16.8
+/0.72
16.6
+/0.56
16.7
+/0.38
100
EL3 - Squid diet
CSI 60
80
Total length (mm)
EL4 - GM
60
40
20
0
* GM: the premium commercial diet
** Salinity at 65 ppt for 60 min (CSI: cumulative stress index)
The number of dead larvae counted every 3 min. At the end of 60 min, the % cumulative mortality
was used as a Cumulative Stress Index (CSI-60). Indication of "condition“ after treatments, stress
resistance, and screening.
17.5
+/0.65
Microdiets from SH with different hydrolysis time
on growth performance of Atlantic cod larvae (50
dph*) in 42 days feeding
Test
diets
SH2
IBL cm
1.76 ±
0.14
SH2-1 1.76 ±
0.14
SH1
1.76 ±
0.14
IBW g
IBCF
BW-g
BL cm
BCF
Survival
SGR %/d
0.040 ±
0.010
0.040 ±
0.010
0.040 ±
0.010
0.72 ±
0.07
0.72 ±
0.07
0.72 ±
0.07
0.526 ±
0.04
0.458 ±
0.02
0.508 ±
0.05
4.07 ±
0.08
3.90 ±
0.06
4.02 ±
0.09
0.77 ±
0.02
0.788 ±
0.02
0.78 ±
0.04
67.7 ±
27.4
65.0 ±8.7
5.99 ±
0.10
5.85 ±
0.03
6.19 ±
0.33
62.9 ±
12.0
SH 2: 2 h hydrolysis at 55C, followed by 30 min at 75C
SH 1: 1 h hydrolysis
SH 2-1: fed 2 h hydrolysate diet for 21 d followed by 1 h diet for 21 d
IBL: initial body length; BW: body wt; BCF: body condition factor;
SGR: specific growth rate
* 30 dph and 20 d acclimation
SDS-PAGE profiles of squid by-product
during hydrolysis
DEGREE OF HYDROLYSIS
25.0
Molecular marker (kDa)
DH (%)
20.0
15.0
10.0
5.0
0
60
120
180
240
300
Time (min)
Figure 3 - Degree of hydrolysis of squid by-product during hydrolysis
1000
Viscosity (cP)
VISCOSITY
Batch_I
800
Batch_II
600
400
200
0
0
60
120
180
240
300
360
Time (min)
Figure 4 - Viscosity changes in squid hydrolysate as affected by hydrolysis time
Surivial, weight, length and specific growth rate
of summer flounder larvae (Paralichthys dentatus) after 22-day
Survival rate (%)
Weight (mg)
Length (mm)
SGR
Artemia
81  2 a
29.8 a
12.2  1.0 a
2.86  0.55 a
Commercial
65  4 b
21.5 4.0 b
11.1  0.8 a
1.39  0.16 b
Squid only
92  3 c
26.2 
Survival and growth of summer flounder larvae (2-wk old)
after
feeding
4.9
ab22-day11.7
 0.8 a
2.23  0.25 ab
100
Surv ival Rate (%); SGR (1-1 0, %)
Diets
Surv iv al Rate
Spe cific Growth Rate
80
60
40
20
0
Arte mia
Comme rc ial
Diets
* SGR = [(ln Wf/Wi)/ days of fe e ding] 100
Squid Hydrolysate
Additional studies to be considered




Feeding trials on additional species
Hydrolysate from finfish-squid combination
Squid hydrolysate for broodstock nutrition
Feed performance test in plant protein-based
aquaculture feeds
Acknowledgements
The present study was supported by the U.S. Dept. of Agriculture,
the Univ. of Rhode Island, Agricultural Experiment Station and the
National Oceanic and Atmospheric Administration (NOAA).
The
information reported in this presentation has been filed for an
invention disclosure (7108PCT; May 17, 2004).