Development of dry fermented sausages rich in docosahexaenoic acid with oil from the microalgae Schizochytrium sp.: Influence on nutritional properties,sensorial quality and oxidation stability

Significant enrichment of dry fermented sausages in n−3 polyunsaturated fatty acids (PUFA) was achieved by incorporating algae oil from Schizochytrium sp. into an emulsion that partially substituted pork backfat from the traditional formulation. Two different levels of substitution were initially tested, both including BHA and BHT as antioxidants: (15% and 25%). A triangular test showed that products with 25% substitution were not sensorially acceptable. Sausages with 15% substitution supplied 1.30 g/100 g product of docosahexaenoic acid (DHA), with an interesting n−6/n−3 ratio of 2.62. No signs of oxidation were detected at the end of the ripening process, with low values of TBARs (<0.2 mg/kg), peroxides (<2 meq O₂/kg fat) and volatile aldehydes. Storage of these sausages under vacuum during 30 days totally guaranteed their stability, whereas after 90 days certain degree of oxidation was detected, only by means of aldehydes analysis. Hexanal, nonanal and dienals increased, whereas no increment in TBARs or peroxides were observed. Aerobical storage of these sausages was not viable due to a high oxidation susceptibility of the new formulation, despite the use of antioxidants.     

Lipid oxidation in n-3 fatty acid enriched Dutch style fermented sausages

Dutch style fermented sausages were manufactured with a substitution of 10%, 15% and 20% of pork backfat by flaxseed oil and canola oil, pre-emulsified with soy protein isolate. The 15% and 20% substitution were also reached by adding encapsulated flaxseed oil and encapsulated fish oil and by adding flaxseed oil, pre-emulsified with sodium caseinate, respectively. The products were sliced, packaged in an oxygen-enriched atmosphere and stored in the dark for 12 weeks at 7°C. No differences were detected in moisture, protein and fat content between control and modified sausages, with the exception of the formulation with sodium caseinate. The PUFA/SFA ratio increased from 0.30 in the control to 0.42-0.48 in the sausages with canola oil and to 0.49-0.71 in the sausages with flaxseed oil. The n-6/n-3 ratio decreased from 11.20 in the control to 6.94-5.12 in the sausages with canola oil and to 1.93-1.05 in the sausages with flaxseed oil. The addition of canola oil and encapsulated flaxseed oil resulted in a comparable shelf life as the control in terms of lipid oxidation. In the samples with addition of pre-emulsified flaxseed oil, especially with sodium caseinate, lipid oxidation clearly increased during storage. Physical and sensory analysis showed that the sausages with encapsulated fish oil and flaxseed oil resembled the control most.     

Nutritional and sensory aspects of light lamb meat enriched in n−3 fatty acids during refrigerated storage

Fatty acid content and sensory characteristics of meat from light lambs fed three diets supplemented with different sources of n−3 fatty acids (fish oil, extruded linseed and extruded linseed plus microalgae) and a control diet during refrigerated storage have been studied. The meat from lambs fed linseed diets had the highest levels of C18:3 n−3, while animals fed fish oil had the highest long-chain n−3 polyunsaturated fatty acids (PUFA). Thus, 100 g of meat from lamb fed the fish oil diet provided 183 mg of long-chain n−3 PUFA, representing 40% of the daily recommended intake. The levels of n−3, n−6 and long-chain n−3 PUFA decreased during a 7-day storage period. The meat from lambs fed the fish oil diet had high scores of fish odour and flavour and rancid odour and flavour and the lowest overall liking. Rancid odour and flavour increased after storage, mainly in supplemented groups.     

Evaluation of lipid oxidation in spaghetti pasta enriched with long chain n−3 polyunsaturated fatty acids under different storage conditions

This study investigated lipid oxidation in spaghetti enriched in long chain n−3 polyunsaturated fatty acids (LC n−3 PUFA) by the addition to semolina of an integrator containing eicosapentaenoic acid and docosahexaenoic acid. Two oxidative parameters were evaluated: peroxide value (PV), to assess primary oxidation and oxidised fatty acids to quantify secondary oxidation products. Functional spaghetti had a shelf life comparable to control pasta. LC n−3 PUFA were not significantly implicated in the onset of oxidation in spaghetti stored under daylight and accelerated oxidation in a laboratory heater. Storage decidedly affected shelf life: PV in functional spaghetti increased from 7.1 to 43.4 meq O₂/kg of fat under light exposure over 12 months, and from 7.1 to 16.2 meq O₂/kg under accelerated ageing at 55 °C for 27 days, reproducing about 18 months at room temperature. Oxidised fatty acids increased in fortified spaghetti from 4.8 to 13.8 g/100 g of fat under light exposure over 12 months and from 4.8 to 7.8 g/100 g of fat at 55 °C in 27 days. The high sensitivity of spectrophotometric and chromatographic analytical methods permitted the evaluation of primary and secondary oxidative derivatives in small amounts of fat.     

Enriching the egg yolk in n − 3 fatty acids by feeding hens with diets containing horse fat produced in Uruguay

The present study was conducted to evaluate the ability of horse fat produced in Uruguay, compared with other lipid sources supplemented in the diet of laying hens, to modify the lipid composition and the n − 6/n − 3 fatty acid ratio of the produced eggs. For this purpose, 60 laying hens (Gallus domesticus) were fed for 30 days with five experimental diets (12 hens/diet) containing 3% sunflower oil (SO), rice oil (RO), beef tallow (BT), pressed-fat (PF), obtained by pressing fat from bovine viscera, and horse fat (HF) obtained from horse bone medulla. Throughout the trial, feeding rate, shell index, weight and total lipid contents of eggs, were not affected by the different diets. Feeding on the SO and RO diets significantly increased the amount of linoleic acid of the egg, although it was lower in the RO than in the SO eggs. Diet BT did not affect the saturated fatty acid content of the yolks. The diets with animal fats containing 18:3n − 3 (diets PF and HF), resulted in a significant increase in the n − 3 fatty acid contents of the eggs, through an increase of linolenic (18:3n − 3) and docosahexaenoic acid (DHA, 22:6n − 3) contents. Eggs from hens fed the HF diet showed increased linolenic acid (46 mg/yolk) and DHA (71 mg/yolk, 1.7% of total fatty acids) contents. These levels were obtained after two weeks of feeding. Moreover, the fatty acid profiles of eggs from treatment HF were not significantly affected by thermal treatment of the yolks. In conclusion, the fat from horse bone medulla, as produced in Uruguay, can be considered as a suitable lipid source for diets of laying hens, to modify the nutritional composition of the eggs in n − 3 PUFA content, especially DHA, and consequently, the n − 6/n − 3 fatty acid ratio.     

Different oilseed supplements alter fatty acid composition of different adipose tissues of adult ewes

Twenty-five mature Small Tail Han ewes were used to investigate the effects of supplemental oilseeds in the diet (sunflower seed, safflower seed, rapeseed, and linseed) on fatty acid composition in different tissues (longissimus lumborum muscle, tail fat, subcutaneous back fat and kidney fat). Averaged over tissue, safflower and sunflower seed was most effective (P < 0.05) in enhancing the concentration of conjugated linoleic acid compared to rapeseed, linseed, and control (1.35% and 1.15% vs. 0.80%, 0.80%, and 0.75%, respectively). Linseed supplemented ewes had lesser n−6/n−3 value (2.48, P < 0.05) compared to sunflower and safflower supplemented ewes (6.12 and 3.90, respectively). Fatty acid composition for most major fatty acids differed among tissues (P < 0.05) but tissue differences varied depending on oilseed supplement (P < 0.05). Proportions of conjugated linoleic acid were greatest in tail fat (1.54% vs. 0.82%, 0.79% and 0.70% for kidney, back, and muscle fat, P < 0.05) as were total unsaturated fatty acids (49.1% vs. 42.4%, 36.7% and 33.4% for muscle, back, and kidney fat, P < 0.05) and tail fat was the most responsive tissue to improvement in fatty acid profile through supplementation. Beneficial fatty acid content of tissues can be increased by oilseed supplementation, but the magnitude of increase varies according to tissue.     

Lipid oxidation of stored eggs enriched with very long chain n−3 fatty acids,as affected by dietary olive leaves (Olea europea L.) or α-tocopheryl acetate supplementation

The antioxidant potential of dietary olive leaves or α-tocopheryl acetate supplementation on lipid oxidation of refrigerated stored hen eggs enriched with very long-chain n−3 fatty acids, was investigated. Ninety-six brown Lohmann laying hens, were equally assigned into three groups. Hens within the control group were given a typical diet containing 3% fish oil, whereas other groups were given the same diet further supplemented with 10 g ground olive leaves/kg feed or 200 mg α-tocopheryl acetate/kg feed. Results showed that α-tocopheryl acetate or olive leaves supplementation had no significant effect on the fatty acid composition and malondialdehyde (MDA) levels of fresh eggs but reduced their lipid hydroperoxide levels compared to controls. Storage for 60 d decreased the proportions of polyunsaturated fatty acids (PUFAs) but increased those of monounsaturated fatty acids (MUFAs) in eggs from the control group, while had no effect on the fatty acid composition of the eggs from the other two groups, which showed decreased levels of lipid hydroperoxides and MDA. Therefore, the very long chain n−3 PUFAs in eggs were protected from undergoing deterioration partly by olive leaves supplementation and totally by α-tocopheryl acetate supplementation. In addition, incorporating tocopherols into eggs might also provide a source of tocopherols for the human diet.     

Production of n − 3 polyunsaturated fatty acid concentrate from sardine oil by lipase-catalyzed hydrolysis

The production of n − 3 polyunsaturated fatty acids (n − 3 PUFAs) concentrate from oil extracted from Pacific sardines (Sardinops sagax) was studied using lipase-catalyzed hydrolysis. Commercially available microbial lipases, from Candida Rugosa (CR), Candida cylindracea (CC), Mucor javanicus (MJ), and Aspergillus niger (AN) were used for enzymatic hydrolyses with extracted sardine oil, run at 37 °C with constant stirring for 1.5, 3, 6, and 9 h. Fatty acid composition analysis by gas chromatography showed that the refined unhydrolyzed oil contained 26.86% of eicosapentaenoic acid (EPA) and 13.62% of docosahexaenoic acid (DHA) (wt/wt%). CR lipase was the most effective in concentrating n − 3 PUFA. Hydrolysis with 250 U CR lipase increased EPA concentration to a relatively constant level of 33.74% after 1.5 h. DHA levels were also significantly increased from 13.62% to 29.94% with 500 U after 9 h. Compared to CR and CC lipases, MJ and AN lipases resulted in low n − 3 PUFA concentration. Triacylglycerol levels decreased significantly as reaction time progressed.     

Seasonal variation in the fatty acid composition of three Mediterranean fish – sardine (Sardina pilchardus), anchovy (Engraulis encrasicholus) and picarel (Spicara smaris)

The total fat contents and the fatty acid compositions of three common Mediterranean fish, namely sardine (Sardina pilchardus), anchovy (Engraulis encrasicholus) and picarel (Spicara smaris) were determined at bimonthly intervals for a one-year period. The purpose of this work was to study the seasonal variation of the fatty acids in the three fish that are some of the best sources of n − 3 fatty acids. The fat and fatty acid content of the investigated fish species show a significant seasonal dependency. Two of the fish (anchovy and picarel) have the highest fat content during the late winter – spring period. On the other hand, sardine shows the highest fat concentrations during the spring-early summer period. The fish that showed the highest variation in fatty acid composition was the anchovy. The sardine was found to be the best source of n − 3 fatty acids during the one-year period (35.35 g/100 g fatty acids). Finally the picarel had the highest oleic acid content (on average, 13.89/100 g fatty acids).     

Fatty acid compositions of selected varieties of Spanish dry ham related to their nutritional implications

Five varieties of Spanish dry cured ham were studied to assess their nutritional value in relation to fatty acids. Ten hams of the Traditional Speciality Guaranteed (TSG) “Jamón Serrano”, and the Protected Designations of Origin (PDO) “Jamón de Teruel”, “Dehesa de Extremadura”, “Jamón de Huelva” and “Guijuelo”, were analysed. Iberian hams (“Dehesa de Extremadura”, “Jamón de Huelva” and “Guijuelo”) were characterised by a lower proportion of saturated fatty acids (SFA) and a significantly higher percentage of monounsaturated fatty acids (MUFA) than white hams (“Jamón Serrano” and “Jamón de Teruel”). The Iberian varieties also showed a high proportion (approximately 50%) of C18:1 n − 9, while “Jamón Serrano” showed the highest percentage of C18:2n−6. The PUFA/SFA (P/S) ratio of the five varieties was ? 0.19, with the highest ratio corresponding to “Jamón Serrano” (0.3). The n−6/n−3 ratio was in the order of 13/1 in “Jamón Serrano” and “Jamón de Huelva”, and ranged from 9.3/1 to 10.3/1 in the other varieties. The most favourable hypocholesterolemic/Hypercholesterolemic (h/H) ratio (?2.5) was found in the Iberian varieties. TSG “Serrano” was shown to supply the lowest percentage of the recommended daily intake of MUFA, the Iberian varieties showed the highest percentage of the daily intake of long-chain PUFA, and PDO “Dehesa de Extremadura” showed the highest percentage of the intake of C18:3n−3. The higher MUFA proportion and h/H ratio observed in the Iberian hams, together with their contribution to the recommended daily intake of fatty acids, would make these products more suitable for healthier diets, although consumption must be recommended in moderation.     

Fatty acids and sensory characteristics of Spanish dry-cured loin enriched in acid α-linolenic and α-tocopherol

The effects of using α-linolenic and α-tocopherol acid-enriched pork on the fatty acids and sensory characteristics of Spanish dry-cured loins were investigated. For the study, five batches of Spanish dry-cured loins were manufactured using pork loin enriched in polyunsaturated n − 3 fatty acids and α-tocopherol. Tissues were obtained from pigs fed on diets with the same ingredients, except for the oil source which corresponded to: [sunflower (C), linseed (L) and linseed and olive (1/1, w/w) (LO)] and two different amounts of α-tocopheryl acetate [20 (C, L and LO) or 200 (LOE and LE) mg/kg diet]. Dry-cured loins with polyunsaturated fatty acid n6/n3 ratios below 4 were obtained from linseed and linseed/olive oil-enriched batches. Dry-cured loin manufactured with pork from animals fed on diets enriched only with linseed oil presented the worst sensory characteristics and higher TBAR values than did dry-cured loins from animals fed on diets enriched with linseed and olive oil and linseed oil plus tocopheryl acetate.     

Effect of pre-emulsified fish oil - as source of PUFA n-3 - on microstructure and sensory properties of mortadella, a Spanish bologna-type sausage

Conventional and low-fat cooked sausages (cursive) were prepared with fish oil as a source of n−3 polyunsaturated fatty acids. Fish oil was incorporated pre-emulsified with caseinates and water. Pre-emulsion was added in sufficient amounts to give fish oil levels of 1–6% in the final product. Lipid oxidation was determined (TBARs index) and instrumental measures of colour and texture were carried out. A sensory analysis was performed in which sensory parameters were evaluated by a hedonic test. There were no significant changes in TBAR values of final products being similar in both control and fish oil added sausages. In general terms, a slight increase in hardness and work of shearing were observed at the higher fish oil levels independently of the fat content. Sausages were analysed after 90 days of storage under refrigeration and similar results were observed. Microstructure of sausages was also studied by scanning electron microscopy (SEM) and a more compact structure was observed when the high levels were assayed. However, all sausages were well accepted when sensory quality was evaluated. Fat composition showed an increase in n−3 PUFA yielding a n−6/n−3 ratio close to 2, which is considered as healthier. For this, the incorporation of fish oil pre-emulsified seems to be a good option to include n−3 PUFA in cooked meat products, obtaining sensorially adequate and healthier stable sausages.     

Additions of caffeic acid,ascorbyl palmitate or γ-tocopherol to fish oil-enriched energy bars affect lipid oxidation differently

The objectives of the study were to investigate the effects of caffeic acid, ascorbyl palmitate and γ-tocopherol on protection of fish oil-enriched energy bars against lipid oxidation during storage for 10 weeks at room temperature. The lipophilic γ-tocopherol reduced lipid oxidation during storage when added at a concentration above 440 μg/g fish oil. However, the best antioxidative effect was observed when it was added at a concentration of 660 μg/g fish oil. In contrast, prooxidative effects were observed when using either γ-tocopherol at concentrations below 220 μg/g fish oil, or the hydrophilic caffeic acid, or the amphiphilic ascorbyl palmitate at concentrations of 75, 150 and 300 μg/g fish oil. Prooxidative effects were observed as an increase in the formation of lipid hydroperoxides and volatile secondary oxidation products, as well as the development of rancid off-flavours. The differences in the efficacies of the three antioxidants examined are expected to be related to their different localisations and mechanisms of action.     

Low-fat meat sausages with fish oil: Optimization of milk proteins and carrageenan contents using response surface methodology

Response surface methodology was used to analyze the effect of milk proteins and 2:1 κ:ι-carrageenans on cooking loss (CL), weight lost by centrifugation (WLC) and texture attributes of low-fat meat sausages with pre-emulsified fish oil. A central-composite design was used to develop models for the objective responses. Changes in carrageenans affected more the responses than milk proteins levels. Convenience functions were calculated for CL, WLC, hardness, and springiness of the product. Responses were optimized simultaneously minimizing CL and WLC; ranges for hardness and springiness corresponded to commercial products (20 g of pork fat/100 g). The optimum corresponded to 0.593 g of carrageenans/100 g and 0.320 g of milk proteins and its total lipid content was 6.3 g/100 g. This formulation was prepared and evaluated showing a good agreement between predicted and experimental responses. These additives could produce low-fat meat sausages with pre-emulsified fish oil with good nutritional quality and similar characteristics than traditional ones.     

Survival of Listeria innocua in dry fermented sausages and changes in the typical microbiota and volatile profile as affected by the concentration of nitrate and nitrite

The involvement of nitrate and nitrite in the formation of N-nitrosamines in foods is a matter of great concern. This situation has led to revise the real amount of nitrate and nitrite needed in meat products to exert proper technological and safety activities, and also to extensive research to find alternatives to their use. The present study addresses the possibility of reducing the ingoing amounts of these additives below the legal limits established by the current European regulations. Different concentrations of nitrate and nitrite were tested on Spanish salchichón-type dry fermented sausages concerning their role in the microbiota and volatile profile. Sausages were manufactured with the maximum ingoing amounts established by the EU regulations (150 ppm NaNO₃ and 150 ppm NaNO₂), a 25% reduction and a 50% reduction; control sausages with no nitrate/nitrite addition were also prepared. The mixtures were inoculated with 5 log cfu/g of Listeria innocua as a surrogate for Listeria monocytogenes. L. innocua numbers in the final product were approximately 1.5 log cfu/g lower in the batch with the maximum nitrate/nitrite concentration when compared to 25 and 50% reduced batches, and about 2 log cfu/g in comparison to the control sausages. The final numbers of catalase-positive cocci were 1 log cfu/g higher in the 50% nitrate/nitrite reduced batch and 2 log cfu/g higher in the control sausages, compared to products manufactured with the maximum nitrate/nitrite concentration. This increase was related to a higher amount of volatile compounds derived from carbohydrate fermentation and amino acid degradation. Sausages with no addition of nitrate/nitrite showed higher amount of volatiles from lipid oxidation. Enterobacteriaceae counts reached detectable values (1-2 log cfu/g) in both nitrate/nitrite reduced sausages and in the control batch, while these organisms were not detected in the batch with the maximum ingoing amount. Nitrate and nitrite exerted a significant effect on the typical microbiota of dry fermented sausages and effectively contributed to control Listeria. These considerations should be taken into account in view of a future restriction in the use of these curing additives.     

Effect of starter culture and fermentation temperature on water mobility and distribution in fermented sausages and correlation to microbial safety studied by nuclear magnetic resonance relaxometry

Water mobility and distribution in fermented sausages produced with differences in pH development as a result of the use of three different starter cultures (T-SPX, F-1, or F-SC-111) and two fermentation temperatures (24 °C, or 32 °C) were studied using low-field proton NMR relaxometry. Changes in the distribution and mobility of water in fermented sausages upon fermentation and drying were detectable by NMR T₂ relaxation, and the progress in the drying process could be followed as a shift towards faster relaxation times. In addition, the distribution of water in the sausages was significantly affected by the pH decline. The sausages were spiked with Listeria monocytogenes, Salmonella, and Escherichia coli VTEC, and partial least squares regressions revealed that 90% of the variation in reduction of Salmonella and VTEC could be explained by the NMR T₂ relaxation decay. Consequently, the study demonstrated that NMR relaxometry is a promising technique for elucidating process parameters and microbial safety in the production of fermented meat products.     

Effects of feeding flaxseed or sunflower-seed in high-forage diets on beef production,quality and fatty acid composition

Yearling steers were fed 70:30 forage:concentrate diets for 205 d, with either grass hay (GH) or red clover silage (RC) as the forage source, and concentrates containing either sunflower-seed (SS) or flaxseed (FS), each providing 5.4% oil to diets. Feeding diets containing SS versus FS significantly improved growth and carcass attributes (P < 0.05), significantly reduced meat off-flavor intensity (P < 0.05), and significantly increased intramuscular proportions of vaccenic (t11-18:1), rumenic (c9,t11-CLA) and n − 6 fatty acids (FA, P < 0.05). Feeding diets containing FS versus SS produced significantly darker and redder meat with greater proportions of atypical dienes (P < 0.05). A significant forage × oilseed type interaction (P < 0.05) was found for n − 3 FA, α-linolenic acid, and conjugated linolenic acid, with their greatest intramuscular proportions found when feeding the RC-FS diet. Feeding GH versus RC also significantly improved growth and carcass attributes, sensory tenderness (P < 0.05) and significantly influenced intramuscular FA composition (P < 0.05), but overall, forage effects on FA profiles were limited compared to effects of oilseed.     

Assessing the quality of organic and conventionally-farmed European sea bass (Dicentrarchus labrax)

The biometric and nutritional traits of European sea bass from organic or semi-intensive conventional production systems at two commercial sizes (small and medium) were compared. The analysis included a total of 80 specimens. The biometric traits and the texture were not affected by the rearing system, whereas they changed significantly with fish size. The fillet fatty acid profile varied significantly, both with rearing system and sea bass size, depending on the fatty acid profile of the diets. The ratio of n − 3 to n − 6 polyunsaturated fatty acids was higher (p < 0.001) in organic than in conventional fish (1.60 vs. 0.54) and in small than in medium-sized sea bass (1.15 vs. 0.98). Near infrared reflectance spectroscopy (NIRS) successfully classified fillets according to sea bass size in both fresh-minced and freeze-dried samples (90% correct classification), whereas it only classified organic vs. conventional sea bass fairly well (65-75% correct classification) for freeze-dried fillets.     

Inhibition of fish oil oxidation and the radical scavenging activity of New Zealand seaweed extracts

This study describes the potential use of New Zealand seaweed extracts as antioxidants in fish oils. The extracts from two brown seaweeds (Ecklonia radiata, Macrocystis pyrifera) and two red species (Champia sp. and Porphyra sp.), were added to hoki (Macruronus novaezelandiae) oil and assessed by production of oxidation products in an elevated temperature (60 °C) storage trial. Antioxidant assays ORAC and DPPH were conducted to elucidate the radical scavenging activity of the seaweed extracts. The results showed that all the lipid-soluble seaweed extracts had some antioxidative capacity, with brown algae generally outperforming red algae species. Extracts from E. radiata performed best with significantly lower primary, secondary and total oxidation products, and higher DPPH radical scavenging ability than the commercial antioxidant, BHT. This study demonstrated the potential of brown algae seaweed extracts, in particular E. radiata, for use as antioxidants in fish oil products.     

Mortadella sausage manufactured with Caiman yacare (Caiman crocodilus yacare) meat,pork backfat,and soybean oil

Mortadellas manufactured using Caiman yacare meat (70%) and pork fat (30%) were reformulated by substituting pork fat with increasing amounts of soybean oil (25%, 50%, and 100%) and evaluated 7, 30, and 60 days post-manufacture. The substitutions resulted in an increase in fat content and in the percentage of polyunsaturated fatty acids (total PUFA; 18:2 n − 6 and 18:3 n − 3), and a reduction in saturated fatty acids (total SFA; 14:0 and 16:0). These alterations reduced (P < 0.01) the indices of atherogenicity and thrombogenicity by 4- to 5-fold in the formulation with 100% soybean oil compared to the original formulation. The levels of TBARS obtained at day 7 were positively correlated (P < 0.01) with 18:2 n − 6 and 18:3 n − 3 (r = 0.83 and 0.84, respectively) and negatively correlated with 17:0 and 17:1 (r = − 0.91 and − 0.89, respectively). All formulations received favourable overall acceptability by the sensory panel.