Optimal Conditions to Remove Chemical Hazards in Fish Protein Isolates from Tilapia Frame Using Response Surface Methodology

Response surface methodology (RSM) was employed to maximize the removal of phospholipids (PLs) and other chemical hazards in tilapia protein isolates made from tilapia frame (TF). CaCl₂ and the ratio of water to minced tilapia frame (W:TF) were the significant variables affecting PLs reduction. The optimum condition for maximal PLs reduction (90.0%) was: 10.25 mM CaCl₂ and a W:TF of 7.8:1, while other variables were fixed at 5 mM citric acid, 60 min incubation, pH 11, and centrifugal speed of 8,000 × g. At these conditions, the great reduction of lipids (93.9%), Hg (97.6%), and As (95.5%), as well as 86.1% of protein recovery, were obtained. Protein isolates with significantly reduced chemical hazards and lipids were successfully prepared from tilapia frames using the alkaline extraction assisted with CaCl₂ and citric acid.     

Antioxidant and Angiotensin-Converting Enzyme Inhibitory Activities of Protein Hydrolysates Prepared from Threadfin Bream (Nemipterus spp.) Surimi By-products

Solid wastes from threadfin bream (Nemipterus spp.) surimi production composed of head and frame were hydrolyzed by various commercial proteases (Alcalase, Flavourzyme, Neutrase, Protamex, papain, and pepsin) to produce protein hydrolysates with bioactive properties. An Alcalase-hydrolyzed sample at 24.4% degree of hydrolysis (DH) displayed the highest antioxidant activity based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) assay, and potassium ferricyanide method. In addition, it showed an inhibitory activity toward angiotensin-converting enzyme (ACE) of 25.5%. Antioxidant activity of threadfin bream by-product hydrolysates increased with hydrolysis time and reached the highest DPPH activity after 6 h, while that hydrolyzed for 3 h showed the highest reducing power based on FRAP and potassium ferricyanide assays. In addition, ACE inhibitory activity was found to be at an optimum after 3 h of hydrolysis. The hydrolysates (1 mg/mL) also retarded oxidation of a linoleic acid emulsion system to a similar extent as 0.1 mg/mL 3-tert-butyl-4-hydroxy anisole (BHA), indicating a potential use in the food system. Protein hydrolysates from threadfin bream surimi by-products could be tailor-made to possess both antioxidant and ACE inhibitory activity through controlling DH of Alcalase-catalyzed reactions.     

Evaluation of the Antioxidant and Antimicrobial Activity of Protein Hydrolysates and Peptide Fractions Derived from Colossoma macropomum and Their Effect on Ground Beef Lipid Oxidation

The objective of this study was to obtain protein hydrolysate from the mechanically separated meat of blackfin pacu to evaluate the influence by ultrafiltration in the antioxidant and antimicrobial activities of the peptide fractions obtained and to apply in ground beef to evaluate the lipid stability. The enzymatic hydrolysis was performed using the enzyme Protamex (pH 7.0, 60°C) for 240 min. The protein hydrolysate was fractionated by ultrafiltration. Then, the antioxidant capacity of the protein hydrolysate and the peptide fractions were evaluated in vitro by the methods of 2,2’-azinobis (3-ethylbenzothiazoline sulfonic acid) radical capture, 2,2-diphenyl-1-picrylhydrazyl radical-scavenging assay, reducing power, and hydroxyl radical scavenging activity. The antimicrobial activity of the samples was evaluated by disc-diffusion against Staphylococcus aureus and Escherichia coli. After evaluation, the peptide fractions did not present higher bioactivities than that shown for the hydrolysate. The protein hydrolysate was applied to ground beef, where the substances reactive to thiobarbituric acid and color were evaluated during 7 days of storage at 4°C. Lipid oxidation was reduced up to 60.9% and there was no modification of the natural coloration. Thus, the protein hydrolysate can be used as an alternative source of antioxidant for the preservation of refrigerated meats.     

Effect of Degree of Hydrolysis and Protease Type on the Antioxidant Activity of Protein Hydrolysates From Cuttlefish (Sepia officinalis) By-Products

We have investigated the antioxidant activities of eight hydrolysates from cuttlefish by-products obtained by treatment with various gastrointestinal proteases (chymotrypsin, trypsin, and crude alkaline enzyme extracts from cuttlefish and sardinelle) and bacterial proteases (Alcalase and crude enzymes from Bacillus pumilus A1, Bacillus mojavensis A21, and Bacillus cereus BG1). The antioxidant activities of the cuttlefish by-products protein hydrolysates (CPHs) were evaluated using various in vitro antioxidant assays, such as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity, reducing power, and total antioxidant capacity. All hydrolysates showed different degrees of hydrolysis (DH) and varying degrees of antioxidant activity. Among the different hydrolysates, cuttlefish crude enzyme hydrolysate exhibited the highest antioxidant activity, followed by sardinelle crude enzyme and Alcalase hydrolysates. Further, CPHs with different degrees of hydrolysis were prepared by treatment with proteolytic enzymes from cuttlefish, sardinelle, and B. mojavensis A21. All hydrolysates showed a greater antioxidative activity as indicated by all the methods considered. In addition, antioxidant activity in hydrolysates was positively correlated with the increase of DH. The results of this study indicated that CPHs might be a good candidate for further investigation in developing new antioxidants.     

Evaluation of the Antioxidant Activity In Vitro and in Hippocampal HT-22 Cells System of Protein Hydrolysates of Common Carp (Cyprinus carpio) By-Product

Fish processing by-products may become more than 50% of the starting material. If mismanaged, these large quantities of discarded fish can create serious pollution problems and can also generate cost associated with their disposal. Enzymatic hydrolysis is one of the techniques that is currently being developed in order to recover and add value to these biomolecules. There is an increasing interest in natural antioxidants that are safer for consumers compared with synthetic antioxidants. In this study, common carp by-product was hydrolyzed using the enzymes Alcalase (A) and Protamex (P) to reach degrees of hydrolysis (DH) of 10 and 15%, respectively. Antioxidant capacity against peroxyl radicals (ACAP); 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging; and the measurement of intracellular reactive oxygen species (ROS) concentration after exposure to common carp protein hydrolysates were investigated. The results revealed that the hydrolysate A15 exhibited significantly (p < 0.05) higher antioxidant activity against the DPPH radical. A15 showed the highest in vitro antioxidant competence against peroxyl radicals, whereas P15 showed the lowest activity against peroxyl radicals (p < 0.05). Hydrolysates having the highest and the lowest in vitro antioxidant activity (A15 and P15, respectively) were selected for the determination of antioxidant activity in the HT-22 cells system. Measurement of intracellular ROS concentration revealed that P15 at the concentration of 1.25 mg/mL significantly (p < 0.05) reduced the intracellular ROS concentration. These results showed that common carp by-product protein hydrolysates are a source of antioxidant peptides with a high potential for food and pharmaceutical industries to develop new nutraceuticals and functional foods.     

Isolation and Characteristics of Anti-Inflammatory Peptides from Enzymatic Hydrolysates of Sandfish (Arctoscopus japonicus) Protein

In the present study, a peptide showing anti-inflammatory activity was isolated from Arctoscopus (A.) japonicus sandfish protein hydrolysate by hydrolysis and analyzed using ultrafiltration (UF), prep-high-performance liquid chromatography (HPLC), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Effects of heat, pH, salt, and intestinal proteases on stability of the isolated anti-inflammatory peptide were determined. The nitric oxide (NO) scavenging activities of the peptide from meat and roe hydrolysates were 18.43 ± 3.21% and 52.35 ± 8.85%, respectively, at a concentration of 0.1 mg/mL. The anti-inflammatory peptide maintained a similar level of NO scavenging activity to the control in the presence of salt, whereas it showed no resistance to heat, pH, and intestinal proteases. These results suggest that the anti-inflammatory peptide derived from A. japonicus could be used for producing functional foods, protein supplements, and pharmaceutical agents. However, it is necessary to find methods for protection against heat, pH, and proteolysis.     

Dietary protein:lipid ratio changes growth,digestive enzyme activity,metabolic profile and haematological parameters in hybrid surubim (Pseudoplatystoma fasciatum × Leiarius marmoratus)

Studies investigating improved protein:lipid ratios based on their effects on growth, nutrient utilization, digestive enzyme activities, blood metabolites and erythogram in tropical carnivorous fish are very scarce. This study evaluated the effect of different protein:lipid ratios on these parameters in the hybrid surubim Pseudoplatystoma corruscans × Leiarius marmoratus. Juvenile fish (8.90 ± 0.94 g initial weight) were fed 3% of total biomass for 60 days using four isoenergetic experimental diets with different protein:lipid ratios (9.00, 4.60, 3.54 and 1.78). Higher growth performance parameters were observed at higher protein:lipid ratios. The activities of intestine total alkaline protease, trypsin and lipase (U/mg protein) were not affected by dietary treatments. Chymotrypsin (U/mg protein) was higher in the 4.60 protein:lipid ratio group. Amylase (U/mg protein) was higher in intermediate groups. Blood glucose, total plasma protein, triglycerides, cholesterol and free amino acids decreased as the protein:lipid ratio decreased. Haematocrit was higher at the 9.00 and 4.60 protein:lipid ratios. Hybrid surubim showed metabolic adaptation to the different protein:lipid ratios tested. These results suggest that the 4.60 protein:lipid ratio showed the best protein‐sparing effect of lipids.     

Production of High Quality Fish Oil by Thermal Treatment and Enzymatic Protein Hydrolysis from Fresh Norwegian Spring Spawning Herring By-Products

There is an increasing demand for omega-3 containing fish oils. By-products from fish fillet production can be utilized for the production of fish oils and be a valuable source of omega-3 for human consumption. The aim of this work was to evaluate industrial processes for production of quality oil for human consumption made from Norwegian spring spawning herring by-products. A mobile production plant was used to compare two industrial processes, thermal treatment (wet rendering) and enzymatic protein hydrolysis, for production of oil from herring by-products immediately after filleting. Results show that high quality herring oil can be produced from fresh by-products. The use of by-products immediately after filleting resulted in a low amount of free fatty acids for all the produced oils (below 0.4%). Thermal treatment at 70°C resulted in an oil with lower oxidation status and higher stability compared to the oils produced by enzymatic protein hydrolysis. Nevertheless, both processing methods gave a crude oil of high quality compared to crude oils on the market.     

Starch and lipid in diets for dourado Salminus brasiliensis (Cuvier 1816): growth,nutrient utilization and digestive enzymes

Carbohydrates and lipids are the major and cheaper non‐protein energy sources for fish diets. This study aimed at determining ideal relationship between dietary starch and lipids and their influence on digestive enzyme profile of juvenile dourado. Trials were set up in a closed‐loop system with controlled temperature, dissolved oxygen, pH and total ammonia. Experimental extruded diets formulated to contain different starch (St) to lipids (Lip) ratio (St : Lip = 0 : 1, 0.3 : 1, 0.7 : 1, 1.0 : 1, 1.4 : 1, 2.0 : 1, 2.3 : 1 to 3.3 : 1) were fed to juvenile dourado (3.34 ± 0.16 g) and stocked into 30 polyethylene tanks (330 L, 30 fish per tank) in a completely randomized design with unbalanced repetition (n = 3 or 4), in two daily meals (1000 and 1530) until satiation, for 95 days. Dietary starch contents higher than 145.2 g kg^‐1 (St : Lip > 1.0 : 1.0) hampered the performance of fish, but, paradoxically, the inclusion of carbohydrates in the diet elicited better utilization of dietary protein and energy by the species. Fish adapted digestive enzyme profile to dietary starch contents through increasing secretion of alkaline proteases and non‐specific lipases, but secretion of amylase was not altered. Physiological responses show the limitation of species on the use of dietary carbohydrates.     

Effect of Dietary Essential Oils Supplementation on Growth Performance,Nutrient Utilization,and Protein Digestibility of Juvenile Gilthead Seabream Fed a Low‐Fishmeal Diet

The present study aimed to assess the efficacy of supplemental plant essential oils (PEOs) on the growth performance and digestive processes of juvenile gilthead seabream. Three experimental diets were tested: (1) a control (CTRL) diet formulated with low marine‐derived protein level (19%); (2) CTRL diet supplemented with a blend of anise, citrus, and oregano essential oils at 1.2 g/kg (diet Phyto C); and (3) CTRL diet supplemented with a similar blend at 0.2 g/kg, but in encapsulated form (diet Phyto E). Triplicate groups of 20 fish (mean initial body weight = 27.9 ± 2.1 g) were fed the experimental diets over 63 d. Results showed that specific growth rate, voluntary feed intake, feed conversion ratio, and protein efficiency ratio were not significantly affected by the two dietary supplements. The commercial blend of PEOs tested here can be successfully incorporated into a low‐fishmeal diet to maximize protein and fat retention and to minimize nitrogenous losses in seabream juveniles. Phyto E presented a decrease in nitrogenous metabolic and fecal losses. The practical implications of including this plant‐based blend in aquafeeds can potentially include faster fish growth and improved production time, but these remain to be tested in a longer experimental trial.     

Dietary monosodium glutamate supplementation during the feed training of pacamã (Lophiosilurus alexandri): Growth performance and intestinal histomorphometry

Feed training of carnivorous fish is a delicate and stressful process. Thus, feed additives that reduce stress and encourage fish consumption could improve training efficiency. Therefore, the aim of this study was to evaluate monosodium glutamate (MSG) dietary supplementation during pacamã (Lophiosilurus alexandri) feed training through growth performance and intestinal histomorphometry. Fish were trained by the method of gradual diet transition, using five diets with increasing proportions of commercial diet and decreasing proportions of gelatin. A quadruplicate experimental design was performed with seven treatments, consisted in diets supplemented with different levels of MSG (0.0; 2.0; 8.0; 16.0; 29.0; 34.0 and 42.0 g/kg). Pacamã (0.17 ± 0.01 g) were distributed in 28 tanks (30 fish per tank) and trained for 41 days with the experimental diets. There were no effects (p > .05) of MSG on growth performance and feed training indices. However, fish trained with diets supplemented with 42.0 g/kg of MSG presented higher number of goblet cells in the anterior portion of intestine (p < .05). These results demonstrate that MSG did not act on growth performance and feed training efficiency of pacamã, and high levels of MSG can cause dietary stress on fish intestinal mucosa.     

Complete replacement of fish meal by porcine and poultry by-product meals in practical diets for fingerling Nile tilapia Oreochromis niloticus: digestibility and growth performance

The apparent digestibility coefficients (ADC) of poultry by-product meal-pet food grade (PBM) and porcine meal (PM) were determined for fingerling male Nile tilapia. ADC for protein were 98.1% and 92.3% in PBM and PM, and 87.1% and 79.4% for energy. PBM and PM were then used as complete replacements for fish meal (FM) in practical diets for Nile tilapia formulated to contain equal digestible protein (300 g kg⁻¹) and energy (16.74 MJ per 100 g) on an as-fed basis. Fingerlings (mean initial weight ±SD, 9.5 ± 0.015 g) were fed for 8 weeks on one of the four diets: FM-, PBM- or PM-based and a commercial feed. The ADC for protein in control and PBM diets (89.7% and 87.9%) were significantly higher than those for the control diet (81.96%). Growth performance and feed utilization were statistically similar between fish fed control and PBM diets, whereas the PM diet exhibited significantly lower performance compared with the control. However, the PM diet showed similar results to the commercial reference diet. Survival and feed conversion ratio were not significantly influenced by replacement of FM with either PBM or PM. The results indicated that PBM and PM can effectively replace FM in practical diets for fingerling Nile tilapia.     

Protein Hydrolysates and Ultrafiltration Fractions Obtained from Krill (Euphausia superba): Nutritional,Functional, Antioxidant,and ACE-Inhibitory Characterization

ABSTRACT

Krill (Euphausia superba) was hydrolyzed by proteolytic enzymes in order to produce multifunctional bioactive peptides, and their functional properties were evaluated. Krill protein hydrolysate (KPH) by pepsin with 4-h hydrolysis showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and angiotensin I converting enzyme (ACE) inhibitory activities. The solubility and foaming properties of KPH were higher than those of the unhydrolyzed krill protein at a wide range of pHs. KPH was further fractionated based on molecular weight. The 1- to 3-kDa peptide fraction exhibited the highest DPPH scavenging activity (IC₅₀ value of 0.5 mg/mL), oxygen radical absorbance capacity (497.39 ± 4.31 µM TE/mg fraction), 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid cation radical scavenging activity (48.41 ± 0.23 µM TE/mg fraction), and reducing power (110.40 ± 2.07 µM TE/mg fraction). However, the < 1-kDa peptide fraction exhibited a higher ACE inhibitory activity than that of other fractions. The 1- to 3- and < 1-kDa peptide fractions are rich in aromatic and hydrophobic amino acids, respectively.     

Protein Hydrolysate from Visceral Waste Proteins of Bolti Fish (Tilapia nilotica): Chemical and Nutritional Variations as Affected by Processing pHs and Time of Hydrolysis

The main objective of the current work was to evaluate the proximate chemical composition and nutritional quality of amino acids of protein hydrolysates from viscera of Tilapia nilotica fish. The results of amino acid composition showed that all hydrolysates contained sufficient and/or excessive amounts of both essential and nonessential amino acids, with appreciable values of essential amino acid/nonessential amino acid ratio (0.77 to 1.59) and essential amino acid/total amino acid ratio (43.35 to 61.35). All protein hydrolysates had a high content of the flavor enhancer amino acids. The lowest values of essential amino acids were found for phenylalanine and methionine with chemical scores ranging from 0.04 to 0.77 and 0.09 to 1.06, respectively, except for 18 h hydrolysates. The protein efficiency ratio had values greatly exceeding 2.0, which is considered a high quality. On the other hand, the levels of sugar and ash contents ranged from 2.69 to 3.32 g L^?1 and 0.48 to 2.1%, respectively, with 9.2 to 14% dry matter. Mineral composition showed that sufficient amounts of calcium, sodium, potassium, magnesium, and iron were present. The results suggest that the studied hydrolysates are appropriate for use in balanced human or animal diets.     

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Analysis of stable carbon and nitrogen isotopes as a tool for assessing the environmental impact of aquaculture: a case study from the western Mediterranean

Stable carbon and nitrogen isotope ratios ( ¹³C/ ¹² C and ¹⁵N/¹⁴ N) of in situ dominant primary producers and consumers were investigated to assess the impact on a natural ecosystem of a land-based fish farm along the south-western coast of Sicily (Italy). The putatively impacted area close to the farm along outfall was compared with other locations at increasing distances from the outfall. Carbon and mainly nitrogen stable isotopes revealed evidence of widespread aquaculture waste in the study site: benthic organisms collected in the impacted area showed more enriched ¹⁵N signatures than those at greater distances from the outfall, suggesting a large contribution of aquaculture waste to the food web. By contrast, both nekton-benthic and pelagic organisms seemed to be less influenced by discharge, probably due to their mobile habit.     

Application of Native Lactic Acid Bacteria (LAB) for Fermentative Recovery of Lipids and Proteins from Fish Processing Wastes: Bioactivities of Fermentation Products

Fermentation using native lactic acid bacteria (LAB) was evaluated for its effectiveness in recovering lipids and proteins simultaneously from freshwater fish visceral waste (FVW). Five different LAB isolated from fish processing waste were employed in a fermentation process that involved 10% (w/w) glucose, 2% (w/w) NaCl, and 10% (v/w) LAB. Cultures evaluated included four native isolates (Pediococcus acidilactici NCIM5368, Enterococcus faecalis NCIM5367, Pediococcus acidilactici FM37, and Pediococcus acidilactici MW2) from FVW with E. faecium NCIM5335 as the reference culture. Fermentation with native LAB resulted in recovery of > 90% oil present in the material as against no recovery in case of raw (unfermented) viscera and resulted in > 50% of degree of hydrolysis of proteins. The fatty acid profile of lipids was not affected by the fermentation process. The fermentation liquor, rich in hydrolyzed protein, exhibited antioxidant as well as antagonistic properties against several bacterial pathogens. The results clearly demonstrate the usefulness of fermentation using native isolates for simultaneous recovery of lipids and proteins from fish processing waste. It also asserts the value of fermentation as an eco-friendly method and aids in minimizing disposal/pollution problems associated with these solid wastes.