Effect of antioxidants on lipid oxidation in herring (Clupea harengus) co-product silage during its production, heat-treatment and storage

Provided high product quality, ensilaging can be used to valorize fish filleting co-products into a silage suitable for food applications. However, a documented challenge for products from hemoglobin-rich fish raw materials is the high susceptibility to lipid oxidation, calling for stabilization by antioxidants. In a comparison among different rosemary-containing antioxidants and isoascorbic acid, we here found that the commercial mixture Duralox MANC-213 (MANC) provided the best protection against peroxide value and 2-thiobarbituric acid reactive substances (TBARS) development during ensilaging of herring filleting co-products (0-7 days, 22 degrees C), and also during subsequent heat-treatment (30 min, 85 degrees C). Increasing MANC concentration from 0.25 and 0.75 to 1.25% lowered TBARS values from 43.53 and 25.12 to 18.04 mu mole TBARS/Kg silage, respectively, after 7 days of ensilaging. During storage at 4 degrees C/22 degrees C in presence of MANC, 1.25% provided the highest protection with 87-90% and 66-73% lower TBARS, at 4 degrees C and 22 degrees C, respectively, at 6 months compared to the controls. At this time point, heat-treated silages had lower protein degree of hydrolysis and free amino acids values than the non-heat-treated one. Regardless of antioxidant addition, total volatile basic nitrogen (TVB-N) formation still increased during the storage, but, overall, TVB-N values in silages were below the acceptable limit of 30 mg TVB-N/100 g fish for human consumption. Together with lipid oxidation data, this suggest that herring silage produced in presence of antioxidants can be used both for high quality feed and food applications

Effect of antioxidants on lipid oxidation in herring (Clupea harengus) co-product silage during its production, heat-treatment and storage

Provided high product quality, ensilaging can be used to valorize fish filleting co-products into a silage suitable for food applications. However, a documented challenge for products from hemoglobin-rich fish raw materials is the high susceptibility to lipid oxidation, calling for stabilization by antioxidants. In a comparison among different rosemary-containing antioxidants and isoascorbic acid, we here found that the commercial mixture Duralox MANC-213 (MANC) provided the best protection against peroxide value and 2-thiobarbituric acid reactive substances (TBARS) development during ensilaging of herring filleting co-products (0–7\ua0days, 22\ua0\ub0C), and also during subsequent heat-treatment (30\ua0min, 85\ua0\ub0C). Increasing MANC concentration from 0.25 and 0.75 to 1.25% lowered TBARS values from 43.53 and 25.12 to 18.04 \ub5mole TBARS/Kg silage, respectively, after 7\ua0days of ensilaging. During storage at 4\ua0\ub0C/22\ua0\ub0C in presence of MANC, 1.25% provided the highest protection with 87–90% and 66–73% lower TBARS, at 4\ua0\ub0C and 22\ua0\ub0C, respectively, at 6\ua0months compared to the controls. At this time point, heat-treated silages had lower protein degree of hydrolysis and free amino acids values than the non-heat-treated one. Regardless of antioxidant addition, total volatile basic nitrogen (TVB-N) formation still increased during the storage, but, overall, TVB-N values in silages were below the acceptable limit of 30\ua0mg TVB-N/100\ua0g fish for human consumption. Together with lipid oxidation data, this suggests that herring silage produced in presence of antioxidants can be used both for high-quality feed and food applications

Understanding the effect of temperature and time on protein degree of hydrolysis and lipid oxidation during ensilaging of herring (Clupea harengus) filleting co-products

The aims of this study were to investigate the effect of temperature, time and stirring on changes in protein degree of hydrolysis (DH), free amino acids (FAA), lipid oxidation and total volatile basic nitrogen (TVB-N) during ensilaging of herring (Clupea harengus) filleting co-products. Results showed that temperature and time, and in some cases the interaction effect between these two factors, significantly influenced all the studied responses. Increasing ensilaging temperature and time from 17 to 37 \ub0C and 3 to 7 days, respectively, increased DH, FAA, and TVB-N content from 44.41 to 77.28%, 25.31 to 51.04 mg/g, and 4.73 to 26.25 mg/100 g, respectively. The lipid oxidation marker 2-thiobarbituric acid reactive substances (TBARS) did not increase with time at temperatures above 22 \ub0C, while 2-pentylfuran increased up to 37 \ub0C. Based on the process parameters and responses investigated in this study, and considering energy requirements, it was suggested to perform ensilaging at ambient temperatures (i.e. around 20 \ub0C) with continuous stirring at 10 rpm for 1-3 days; the exact length being determined by the desired DH

High-throughput sequencing of gut microbiota in rainbow trout (Oncorhynchus mykiss) fed larval and pre-pupae stages of black soldier fly (Hermetia illucens)

Black soldier fly (Hermetia illucens) meal is a potential alternative to fishmeal and plant proteins in diets for farmed fish since it can be produced on organic waste substrates, requires little energy and water inputs and contains high levels of essential amino acids. Recent studies have partially replaced fishmeal with black soldier fly meal, however, research on their impact on gut microbiota of fish is limited. In a five week experiment, juvenile rainbow trout (Oncorhynchus mykiss) were fed either a reference diet based on fishmeal or three diets with 30% inclusion of black soldier fly meals in the form of pre-pupae, larvae or defatted-larvae. The combined luminal content and mucosa were collected from the distal intestine of three fish per tank with four tanks per diet (n = 12) and 16S rRNA gene amplicons were sequenced using the Illumina MiSeq platform. Feeding the insect-based diets increased the alpha-diversity of bacteria and abundance of lactic acid bacteria, which may be due to the addition of dietary chitin. Compared with fishmeal, feeding insects resulted in higher abundance of phyla Firmicutes and Actinobacteria with lower abundance of Proteobacteria. Fish fed the full-fat meals had higher abundance of Corynebacterium that was attributed to its ability to produce lipase and the high content of dietary lipids as a substrate. Bacillaceae was increased in fish fed both larvae diets and unchanged in the pre-pupae diet, which indicated that life-cycle stage of the insect influenced the gut microbiota. Based on these results, we found that feeding black soldier flies increased diversity and altered the composition of gut bacteria of rainbow trout, which were further influenced by life-cycle stage and lipid content of the insect meal

Comparative survival and growth performance of European lobster Homarus gammarus post larvae reared on novel feeds

One approach to ongrow juvenile European lobster, Homarus gammarus, is to utilize land based rearing systems, incorporating automated feeding, individual culture and provision of stable pelleted feeds, preferably using sustainable ingredients. We initiated three feeding experiments to investigate the general suitability of ingredients produced from seafood by‐products as novel feeds for H. gammarus , in terms of promoting survival, development and growth of post‐larval lobsters from post‐larvae (PL) stage IV to the first juvenile stage (stage V). The first experiment was designed to screen an array of candidate, locally produced, novel protein sources on growth performance parameters. This initial experiment revealed that PL reared on a raw (i.e. wet, unprocessed shrimp) feed used as a reference showed superior performance to those reared on experimental feeds containing fishmeal, herring protein isolate or mussel meal; however, a novel type of shrimp meal, produced by flocculation from waste water, promoted the best PL performance of any experimental feed. A second experiment was designed to test the effect of drying method and to optimize the form of a wet shrimp reference feed used by lobster hatcheries. This showed that the performance of PL reared on experimental freeze‐dried shrimp feed was not significantly different to those reared on the wet, unprocessed shrimp used as a reference feed. However, lobsters offered experimental oven‐dried shrimp feed (with or without an immune supplement) resulted in significantly lower survival or growth performance. A third and final experiment was designed in an attempt to improve a candidate herring‐based protein source, by supplementing with nutrients found in shrimp. However, the results showed that PL reared on the wet reference shrimp feed still showed superior growth and survival than those reared on a herring feed alone, or supplemented with additives found in shrimp meal (either glucosamine, astaxanthin or both supplements combined). The high survival and growth, low incidence of moulting problems and high availability of waste shrimp material, suggest that non‐heat‐treated shrimp products are a promising feed ingredient for post‐larval European lobsters

Marine yeast (Candida sake) cultured on herring brine side streams is a promising feed ingredient and omega-3 source for rainbow trout (Oncorhynchus mykiss)

A major challenge for the aquaculture industry is the supply of sustainable feeds. A promising model to achieve this is to utilize circular flows where feed ingredients, such as single cell protein, are cultivated using side streams of the food industry. The aim of this study was to evaluate the marine yeast Candida sake, produced on herring brine side streams, as a source of protein and immune stimulant in feed for salmonid fish. The dry C. sake product contained 54% protein (3.3% lysine and 0.8% methionine) and 13% lipids (1.1% eicosapentaenoic, EPA, and 1% docosahexaenoic acid, DHA). Four experimental diets were designed and tested in a 9-week feeding trial using juvenile rainbow trout (Oncorhynchus mykiss). A control diet containing both fish and plant-based ingredients constituted the base feed to which 20% (to evaluate effects on digestibility, growth and intestinal physiology), 20% heat-treated (to evaluate effects of downstream processing) and 3% (to evaluate immune stimulatory properties, replacing 3% soy protein concentrate) C. sake was added. The apparent digestibility coefficient of C. sake for protein, fat and gross energy was above 80%, and for amino acids above 90% regardless of treatment, suggesting a high bioavailability of C. sake. All three yeast containing diets performed equally to the control regarding specific growth rate, feed conversion ratio and functional intestinal health. These results suggest that C. sake is a promising alternative protein source for circular feeds in the salmonid industry. The presence of EPA and DHA represents an added value. The heat treatment increased the apparent digestibility coefficient of dry matter by 8% but decreased amino acid digestibility by on average 3%, indicating that heat treatment may not be the optimal downstream processing technique. Furthermore, the inclusion of 3% C. sake increased the intestinal lamina propria width and TGF-β transcription, indicating an immune stimulating effect. Future research is needed to understand these immune modulatory effects of C. sake supplementation

Marine yeast (Candida sake) cultured on herring brine side streams is a promising feed ingredient and omega-3 source for rainbow trout (Oncorhynchus mykiss)

A major challenge for the aquaculture industry is the supply of sustainable feeds. A promising model to achieve this is to utilize circular flows where feed ingredients, such as single cell protein, are cultivated using side streams of the food industry. The aim of this study was to evaluate the marine yeast Candida sake, produced on herring brine side streams, as a source of protein and immune stimulant in feed for salmonid fish. The dry C. sake product contained 54% protein (3.3% lysine and 0.8% methionine) and 13% lipids (1.1% eicosapentaenoic, EPA, and 1% docosahexaenoic acid, DHA). Four experimental diets were designed and tested in a 9-week feeding trial using juvenile rainbow trout (Oncorhynchus mykiss). A control diet containing both fish and plant-based ingredients constituted the base feed to which 20% (to evaluate effects on digestibility, growth and intestinal physiology), 20% heat-treated (to evaluate effects of downstream processing) and 3% (to evaluate immune stimulatory properties, replacing 3% soy protein concentrate) C. sake was added. The apparent digestibility coefficient of C. sake for protein, fat and gross energy was above 80%, and for amino acids above 90% regardless of treatment, suggesting a high bioavailability of C. sake. All three yeast containing diets performed equally to the control regarding specific growth rate, feed conversion ratio and functional intestinal health. These results suggest that C. sake is a promising alternative protein source for circular feeds in the salmonid industry. The presence of EPA and DHA represents an added value. The heat treatment increased the apparent digestibility coefficient of dry matter by 8% but decreased amino acid digestibility by on average 3%, indicating that heat treatment may not be the optimal downstream processing technique. Furthermore, the inclusion of 3% C. sake increased the intestinal lamina propria width and TGF-beta transcription, indicating an immune stimulating effect. Future research is needed to understand these immune modulatory effects of C. sake supplementation

Effects of dietary yeast inclusion and acute stress on post-prandial whole blood profiles of dorsal aorta-cannulated rainbow trout

Yeast is a potential alternative to fish meal in diets for farmed fish, yet replacing more than 50 % of fish meal results in reduced fish growth. In a 4-week experiment, 15 rainbow trout (Oncorhynchus mykiss) were cannulated and fed three diets each week: 30 % fish meal as a control (FM); 60 % replacement of fish meal protein, on a digestible basis, with Saccharomyces cerevisiae (SC); and 60 % replacement with Wickerhamomyces anomalus and S. cerevisiae mix (WA). Blood was collected at 0, 3, 6, 12 and 24 h after feeding. In the final week, fish were exposed to a 1-min netting stressor to evaluate possible diet-stress interactions. Significant increases in pH, TCO2, HCO3 and base excess were found after fish were fed the SC and WA diets compared with FM, which elevated blood alkaline tides. Yeast ingredients had lower buffering capacity and ash content than fish meal, which explained the increase in alkaline tides. In addition, fish fed the WA diet had significantly reduced erythrocyte area and fish fed SC and WA diets had increased mean corpuscular haemoglobin levels, indicating haemolytic anaemia. Higher levels of nucleic acid in yeast-based diets and potentially higher production of reactive oxygen species were suspected of damaging haemoglobin, which require replacement by smaller immature erythrocytes. Acute stress caused the expected rise in cortisol and glucose levels, but no interaction with diet was found. These results show that replacing 60 % of fish meal protein with yeasts can induce haemolytic anaemia in rainbow trout, which may limit yeast inclusion in diets for farmed fish

Nutrition of Arctic charr (Salvelinus alpinus) and Eurasian perch (Perca fluviatilis) and evaluation of alternative protein sources

The aim of this thesis was to estimate the dietary lysine requirement of Eurasian perch (Perca fluviatilis), to study the digestive enzyme activity in perch and Arctic charr (Salvelinus alpinus) and the utilization of meals of microbial and mussel meals. Juvenile Eurasian perch were fed semi-purified diets containing graded levels of lysine (12.2-24.3 g kg-1 dry matter (DM), (3.0-6.0 g/16 g nitrogen (N)) to estimate the lysine requirements. Fish fed diets containing 18.3-24.3 g lysine kg-1 DM had higher final body weight, weight gain, protein gain and specific growth rate than fish fed diet containing 12.2-17.3 g lysine kg-1 DM. The results indicate that the lowest lysine level required for optimal growth performance in Eurasian perch is 18.3 g lysine kg-1 DM (4.5 g/16 g N). The activity of digestive enzymes was studied in farmed Eurasian perch and Arctic charr. There were no differences in the activity of lipase, α-amylase and disaccharidases between slow and fast growing perch, or between perch of different age. The total lipase activity was higher than total carbohydrase activity in both species. Perch had a higher total activity of carbohydrases than charr, suggesting a greater capacity for digesting carbohydrates, especially starch. In Artic charr, the apparent digestibility coefficients (ADC) of dry matter (DM), amino acids (AA) and energy was lower for intact Saccharomyces cerevisiae than for extracted S. cerevisiae, Rhyzopus oryzae and Mytilus edulis, whereas the ADC of crude protein (CP) did not differ between the ingredients. With the exception for arginine and histidine, differences were found in ADC of indispensable amino acids (IAA) between the test ingredients. In Eurasian perch, there were no significant differences in the ADC of DM, CP, IAA and energy between the test ingredients. The results indicate that 40% of CP derived from fish meal in diets for Arctic charr can be substituted with blue mussel (M. edulis) meal or intact S. cerevisiae yeast without any negative effect on growth performance and nutrient utilization