Progress 04/01/01 to 09/30/04
Outputs
Squid (Illex illecebrosus and Loligo pealei) processing waste accounts for approximately 40% of the incoming raw squid weight and approximately 8 million pounds are generated annually in Rhode Island and New Jersey. The waste largely consists of head, fin, and viscera along with unclaimed mantles and tentacles. The protein content of the waste is high enough for proteolytic hydrolysis (enzymatic digestion) to generate large amounts of peptides and free amino acids responsible for feed attraction and growth stimulation. Protein and lipid constitute 11-14% and 2-3% of byproducts, respectively, and the oil contains 11.16% EPA and 24.45% DHA . The most viable approach to full utilization of this waste would be a bioconversion into hydrolysate as a nutrient additive to aquaculture feeds. In addition, squid hydrolysate can be used as a feed attractant owing to its high content of betaine and other attractant amino acids. Hydrolysis by its endogenous proteolytic enzymes makes
the process cost effective. The resulting squid hydrolysate is found to have nutrition enhancing and attractant properties desirable for larval and brood fish. Our work included optimization of hydrolysis process, chemical characterization of hydrolysate, formulation and pilot plant production of squid hydrolysate-based microparticulated starter feed, and lab scale feeding trials on trout fingerlings, salmon fry and summer flounder larvae. Field feeding studies were conducted on Atlantic cod larvae and brood black seabass and cod at a commercial hatchery. Commercial scale production of squid hydrolysate and microparticulated starter feed were successfully accomplished. In one batch for production of squid hydrolysate, 1200 lb squid processing waste was chopped and hydrolyzed for 2 h at 55C and finished at 75C for 30 min, filtered through a series of vibrating screens, and concentrated in a vacuum evaporator at 48C from 87% to 75-78% moisture. The resulting squid hydrolysate can be
used as is for brood fish as a nutrition enhancer and attractant or for larval fish feeds as a highly digestible protein source. The formulated feed mix was homogenized in a two-stage homogenizer and dried in flake using a drum dryer at below 100C. Flakes were ground and sieved to desired sizes. The field feeding trials on Atlantic cod larvae and brood black sea bass as well as a lab feeding trial on summer flounder demonstrated that squid hydrolysate diet exhibited a superior feeding performance compared to commercial premium class diets in terms of survival and growth as well as salinity stress tolerance. The present form of diet significantly shortened the transition period from rotifer to a formulated diet partially replacing Artemia live diet, but did not achieve 100% replacement. With further improvement of formulation in digestibility and physical properties, the 100% replacement can be achieved. Additional field feeding trials on tuna, shrimp and European sea bream are
currently underway. With conclusion of successful feeding trials, a business plan has been prepared for a commercial production and marketing of squid hydrolysate and starter diet.
Impacts
Successful feeding trials using processed squid waste were performed on various larval fish species.The squid hydrolysate was also attractive to brood fish. This work supports the potential for commercial production of hydrolysate and premium starter diet from squid processing waste. This will support the local economy while alleviating waste disposal problems.
Publications
- No publications reported this period
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Progress 01/01/03 to 12/31/03
Outputs
A commercial scale production of squid hydrolysate was carried out at a commercial processing facility. Squid by-product was chopped in a homogenizer, transferred into a 300-gal steam-jacketed vessel, and heated to 52-55C. The hydrolysis was terminated at the viscosity of near 200 cP,immediately followed by heating to 75C for 30min. The resulting pasteurized hydrolysate was successively passed through vibrating screens. The filtrate was concentrated in a vertical vacuum evaporation system with forced circulation at 48C and 28in Hg vacuum until the concentration of hydrolysate increased from 12 to 25 Brix or the moisture content from 87.05 to 78.18%. Yield of the final product was found to be 51.25%. The experimental microparticle diet for the larval summer flounder (Paralichthys dentatus) feeding trial was formulated with squid hydrolysate or squid byproduct-herring (7:3) hydrolysate with various ingredients, including crude salmon oil, vitamin and mineral premix,
algae (equal amount of spirulina:chlorella), baker yeast and starch. All ingredients were pre-mixed in a Hobart mixer in the specified order and homogenized by passing through a two-stage Gaulin homogenizer for proper emulsification and encapsulation of water-soluble ingredients. The resulting homogenate was drum dried at a rate of 1 kg product/hr. The dry sheets were ground and screened through a series of sieves: 500-300-150 micrometer. Particles of two sizes, 150-300 and 300-500 micrometer, were collected for proximate composition and feeding trial. A feeding trial of summer flounder larvae was conducted using two squid hydrolysate-based experimental diets, a live feed (Artemia), and a commercial starter feed. Summer flounder larvae of 14 dph were randomly arranged into 13 aquaria (21 L, 48 larvae each) filled with 11.5 L seawater at 18.5+/-1.5 C, pH 7.8-8.0, salinity 28-30 g /L in triplicate except for the control (no food given). Feeding was carried out manually five times daily
to satiation at a daily dose (20% of the fish weight). An initial group of 10 larvae was sampled at the start of the experiment for the measurements of the weight and standard length. Upon the completion of 22-day feeding, the weight and length of each larva were recorded and survival rate and specific growth rate calculated. The stomach color of fish larvae fed with squid hydrolysate-based diets were gradually changed from orange (Artemia) to slight brown during the first three-day feeding indicating the acceptance of the squid hydrolysate diet. The survival rate (91.67+/-2.95%) and SGR (2.23) of larvae fed with squid hydrolysate were significantly (p < 0.05) higher than those fed squid-herring hydrolysate and commercial diets. However, the SGR was not different than that of Artemia (2.86). The commercial diet showed least survival (65.28+/-4.34%) and SGR (1.39). All larvae of control group(starvation) were dead at day 17. The results clearly indicate that the squid hydrolysate
microparticale diet can be used effectively for weaning of summer flounder larvae in place of Artemia.
Impacts
With success of further feeding trials on other commercially important species, high-value starter diets can be produced and commercialized for profit from the squid processing byproduct, presently discarded in large quantities.
Publications
- Lian P.Z. and Lee C.M. 2003. Characterization of squid hydrolysate for its potential as aquaculture feed ingredient. Proceedings of the Trans-Atlantic Fisheries Technology (TAFT) 2003 Conference. Paper No. L 74: p. 379-380. (Abstract)
- Lian P.Z., Lee C.M., and Park E. 2003. Characterization of squid processing wastes (Loligo pealei) during hydrolysis and its attracting properties to trout fingerling. The 2003 Annual IFT Meeting Technical Program Abstracts. Abstract No. 102-1, p. 251.
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Progress 01/01/02 to 12/31/02
Outputs
In the 2nd phase of the study, we investigated feed attractability and conversion of squid hydrolysate as well as survival rate on trout fingerlings and salmon starters and juveniles. Other activities included commercial scale production trial, development of preservation technique, and starter diet preparation with a hydrolysate emulsion. Squid hydrolysate produced at 2 hr-hydrolysis showed the stronger attractability (21 out of 25 fish) than control (4/25) and 3 hr-hydrolysate (10/25) when was used a hydrolysate-soaked cotton ball retained in the perforated plastic golf ball. This may be attributed to increases in attractant amino acids, gly, ala, and val by 275, 210, and 285%, respectively. In Atlantic salmon juvenile feeding, diets were prepared with fish meal replacement at 0, 5 and 10% on a protein weight basis. A higher survival rate (77.5% over 65% control) of the diet with 10%-squid hydrolysate replacement, and a higher feed efficiency ratio (1.62 ,b 0.11
over 1.34 ,b 0.02 control) with 5% replacement were observed. The effect of squid hydrolysate as an attractant and growth stimulation on Atlantic salmon starter was studied using a commercial salmon starter diet spray-coated with 5% and 10% (on a diet weight basis) of liquid squid hydrolysate and oil mixture (8:2) in the form of emulsion. Samples coated with squid hydrolysate emulsion had a significantly less leaching (80% over 100% control) during the first 30 min, indicating a barrier property for nutrient retention and pollution control in effluent. Upon 7-week feeding salmon sac fries (50 fish per 110 gal aquarium), the food conversion ratio (FCR) and daily weight increase ratio (DWR) in diet coated with 5% of squid hydrolysate were 0.96 and 2.81, respectively, compared to control (1.12 and 2.56). A large scale hydrolysis (1522 lb raw material) was successfully carried out under the previously established hydrolysis condition (55 C for 2 hr) at a commercial processing facilities
equipped with homogenizer, 300 gal-reaction vat, vibrating screens and falling-film evaporator (48 C/28 in Hg). Hydrolysis was terminated when the viscosity reached 200 cP, immediately followed by heating to 75C for 30 min. The filtrate was concentrated from 12 to 25 Brix or moisture content from 87.05 to 78.18%. Yield of the final product was found to be 51.25% on a wet weight basis. Concentrated squid hydrolysate was effectively preserved with 85% H3PO4 at 1.75% (v/v). It was necessary to use byproducts as fresh as possible since a high level ammonia was formed during a two-day storage from 136 to 1071 mg-N/100 g solids. Our focus of the feeding study will be on a starter diet since the starter diet will bring a greater return compared to juvenile or grower diets because of its considerably higher market price per unit weight, where squid hydrolysate will be used as a specialty ingredient in partial replacement of fish meal. Additional species to be tested with squid hydrolysate
will include summer flounder and Atlantic cod.
Impacts
With renewed efforts in mariculture of Atlantic cod and summer flounder, the strong feed sttractant and stimulant properties will make squid hydrolysate a good candidate for starter diet ingredient. If it proves to be effective, the potential of squid hydrolysate for a high-value product can be commercially realized.
Publications
- No publications reported this period
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Progress 10/01/01 to 03/31/02
Outputs
The processing of the whole squid yields 40 - 50% waste with low fat, high protein and endogenous enzymes which is ideal for the production of hydrolysate without commercial enzymes added. The present investigation consists of construction of pilot plant scale reaction system, determination of an optimum hydrolysis condition, hydrolysate production for aquaculture feed preparation with cost analysis, and feed quality analysis by feeding trial. This progress report includes construction of pilot plant reaction system and determination of an optimum hydrolysis condition. The reaction vessel was constructed with a stainless-steel reaction chamber (15 gal) housed in a retort vessel which provides a heating medium. The temperature of the reaction medium (squid homogenate) is regulated by hot water whose temperature is controlled by steam injection. The filtered hydrolysate (87% moisture) was concentrated using a concentrator to 71% or lower depending upon the solid content
requirement for feed formulation. Currently, a hot-water jacketed cooker (40 gal) is being used as a batch concentrator. Both retort vessel and concentrator utilize a temperature-controlled hot water circulation system. A steam injection regulator has been installed for automatic control of the temperature of the heating medium. Hydrolysis was carried out for 0, 0.5, 1, 1.5, 2, 3, 4 and 5 hr and analyzed for changes in the degree of hydrolysis (DH), viscosity, protein and peptide profiles, fatty acid profile, and proximate composition. The moisture, lipid, ash and protein contents in the raw squid processing waste were approximately 85.3-86.7%, 1.8-2.3%, 1.2-1.4% and 10.15-10.75%, respectively. The DH value markedly increased from 10.17-10.49 to 16.5-18.7 upon 2 hr hydrolysis, where the initial high DH value reflects the rapid initiation of hydrolysis upon mechanical homogenization prior to the heat-assisted reaction. Viscosity of the hydrolysate exponentially decreased from 847-900
to 13-15 cps. No further marked changes in DH and viscosity after 2 hr hydrolysis. The SDS-PAGE revealed that 2-hr hydrolysis with a DH value of around 17 yielded peptides as the major fraction with a small fraction of partially hydrolysed proteins which is believed to be a contributing factor to an optimum nutrition for fish growth. The progress of hydrolysis was successfully monitored by a viscosity measurement. In the 2nd phase of the study, a sufficient amount of hydrolysate will be produced for the preparation of starter feeds for salmon, summer flounder and cod. Feeding trials will be conducted for the determination of the growth and survival rates.
Impacts
A new specialty aquafeed ingredient designed for starter and juvenile will be produced from otherwise unusable squid processing by-products which account for 40-50% of the incoming whole squid weight. It is our hope that this squid hydrolysate will serve as a growth and immune stimulant so that the feed manufacturers will use it as a specialty ingredient. This will lead to a full commercail use of this unwanted by-products for a better economic return with elimination of waste disposal problems.
Publications
- No publications reported this period
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