Effects of −1.5 °C Super-chilling on quality of Atlantic salmon (Salmo salar) pre-rigor Fillets: Cathepsin activity,muscle histology,texture and liquid leakage

The aim of this study was to evaluate the impact of super-chilling on the quality of Atlantic salmon (Salmo salar) pre-rigor fillets. The fillets were kept for 45 min in a super-chilling tunnel at −25 °C with an air speed in the tunnel at 2.5 m/s, to reach a fillet core temperature of −1.5 °C, prior to ice storage in a cold room for 4 weeks. Super-chilling seemed to form intra- and extracellular ice crystals in the upper layer of the fillets and prevent myofibre contraction. Lysosome breakages followed by release of cathepsin B and L during storage and myofibre–myofibre detachments were accelerated in the super-chilled fillets. Super-chilling resulted in higher liquid leakage and increased myofibre breakages in the fillets, while texture values of fillets measured instrumentally were not affected by super-chilling one week after treatment. Optimisation of the super-chilling technique is needed to avoid the formation of ice crystals, which may cause irreversible destruction of the myofibres, in order to obtain high quality products.     

Atlantic salmon (Salmo salar) muscle structure integrity and lysosomal cathepsins B and L influenced by dietary n-6 and n-3 fatty acids

This study had two main objectives: first, to evaluate the impact of different types and levels of dietary n-6 and n-3 fatty acids (FAs) on Atlantic salmon muscle structure integrity; second, to highlight a possible role of lysosomes and lysosomal degrading enzymes, cathepsins, in fish muscle structure integrity, in relation to dietary fatty acids. Four groups of Atlantic salmon (90 g starting weight) in fresh water tanks were fed one of four diets containing 23% crude lipids, with 100% of the added oils as either fish oil (FO), rapeseed oil (RO), eicosapentaenoic acid (EPA) enriched-oil or docosahexaenoic acid (DHA) enriched-oil. The RO diet was characterised by low levels of EPA + DHA (10% of total FAs), whereas the EPA and DHA diets were characterised by very high levels of EPA + DHA (>50% of total FAs). Fatty acid composition of the muscle crude lysosomal fraction (CLF) generally reflected the diets. Salmon fed the RO diet presented a muscle CLF FA composition close to the one of the FO group, showing moderate PUFA levels, and comparable cathepsin B and cathepsin L activities, relative gene expression of cathepsin B and cathepsin L in the muscle and rate of myofibre–myofibre detachments post-mortem. Salmon fed the EPA and DHA-enriched-oil diets presented a fairly similar muscle CLF FA composition, but different from the FO and RO groups. In the EPA and DHA groups, the percentage of PUFAs in the muscle CLF, the rate of myofibre–myofibre detachments and the relative gene expression of cathepsin B were higher than in the FO and RO groups. Cathepsin B and cathepsin L total activities in the muscle were however lower in the EPA and DHA groups 0 h post-mortem. Dietary lipids influenced the level of lysosomal degrading enzyme activity cathepsin B and cathepsin L as well as the relative gene expression of cathepsin B. Feeding Atlantic salmon with rapeseed oil and extreme levels of EPA + DHA highlighted the impact of fatty acid composition of the diet on salmon muscle integrity and the complexity of the process involving muscle lysosomes and cathepsins in relation to these dietary fatty acids.     

Relevance of calpain and calpastatin activity for texture in super-chilled and ice-stored Atlantic salmon (Salmo salar L.) fillets

The aim of the present experiment was to measure the protease activities in ice-stored and super-chilled Atlantic salmon (Salmo salar) fillets, and the effect on texture. Pre-rigour fillets of Atlantic salmon were either super-chilled to a core temperature of −1.5 °C or directly chilled on ice prior to 144 h of ice storage. A significantly higher calpain activity was detected in the super-chilled fillets at 6 h post-treatment compared to the ice-stored fillets and followed by a significant decrease below its initial level, while the calpastatin activity was significantly lower for the super-chilled fillets at all time points. The cathepsin B + L and B activities increased significantly with time post-treatment; however, no significant differences were observed at any time points between the two treatments. For the ice stored fillets, the cathepsin L activity decreased significantly from 6 to 24 h post-treatment and thereafter increased significantly to 144 h post-treatment. There was also a significantly lower cathepsin L activity in the super-chilled fillets at 0 h post-treatment. No significant difference in breaking force was detected; however, a significant difference in maximum compression (F_max) was detected at 24 h post-treatment with lower F_max in the super-chilled fillets. This experiment showed that super-chilling had a significant effect on the protease activities and the ATP degradation in salmon fillets. The observed difference in F_max may be a result of these observed differences, and may indicate a softening of the super-chilled salmon muscle at 24 h post-treatment.     

Myosin heavy chain degradation during post mortem storage of Atlantic cod (Gadus morhua L.)

Post mortem proteolytic degradation of fish fillets leads to textural changes like muscle softening and gaping. In this study proteolytic degradation of myosin heavy chain (MHC) was monitored during storage of muscle and of isolated myofibrils at different temperatures and pH-values by the use of MHC-specific antibodies. The ability of cathepsin D to associate to myofibrillar proteins was also studied. Muscle stored at 6 °C and isolated myofibrils stored at 0 °C, 6 °C and 20 °C were degraded at pH 6.3 or lower. Cathepsin D could be found associated with extensively washed myofibrils. Inhibition of cathepsin D during storage affected the observed MHC-degradation at pH 5.5, but not at pH 6.3. This indicates that cathepsin D to a less extend than formerly believed, is responsible post mortem degradation of MHC.     

Effects of pre-slaughter stress on proteolytic enzyme activities and muscle quality of farmed Atlantic cod (Gadus morhua)

Farmed Atlantic cod were subjected to a combination of stressors in a holding tank before being killed, pre rigor filleted and stored in ice. At slaughter, a higher level of stress was confirmed by blood physiology analyses. This was further associated with significantly reduced muscle pH and somewhat elevated muscle collagenase-like activity in the stressed fish, whereas no differences in cathepsin-like activities were found. After 5 days of iced storage, the stressed fish had significantly lower water holding capacity, reduced hardness and yellowish colour compared to the control group, and no differences in the other parameters investigated. Independent of pre-slaughter stress, the activities of cathepsin B- and B/L-like enzymes increased and activities of cathepsin D/E- and collagenase-like enzymes decreased with storage.     

Correlation between astaxanthin amount and a* value in fresh Atlantic salmon (Salmo salar) muscle during different irradiation doses

The amount of astaxanthin and a* value changes in fresh Atlantic salmon light and dark muscle during cold storage was studied for different e-beam doses (0, 1, 1.5, 2 and 3 kGy). Astaxanthin (mg/kg muscle) and a* value decreased with increasing irradiation dose for both fresh light and dark muscle. The level of irradiation dose gave high correlation between a* value and amount of astaxanthin. The reason for the change in colour or decrease in a* value of Atlantic salmon during irradiation could be due to the destruction of astaxanthin. The amount of astaxanthin and a* value of 1 kGy treated salmon fillets were not significantly different (p > 0.05) from that of the control but significantly different (p < 0.05) from other irradiation treatments. The colour (a* value) of salmon muscle was related to the content of astaxanthin, which decreased as irradiation increased. The amount of astaxanthin in light muscle was three to five times greater than dark muscle. This study demonstrated that irradiating salmon fillets at 1 kGy, can be successfully used and leads to no significant change in colour and amount of astaxanthin.     

Pre-rigor filleting and brining of farmed cod: Influence on quality and storage stability

Pre-rigor filleting as well as brining with a mixture of phosphates and salt could influence several quality parameters of cod fillets. The aim of this work was to evaluate which of these processes that affected selected quality parameters of farmed cod the most. The analyses were performed at several points during storage, enabling that the effects of time of filleting, brining and storage could be evaluated simultaneously.Pre- and post-rigor non-brined fillets had similar and significant weight loss during storage. The weight increase of pre- and post-rigor fillets due to brining were 2% and almost 10%, respectively, and brining significantly reduced the weight loss during storage. Liquid loss and total viable count (TVC) were significantly influenced by rigor state at time of filleting, brining and storage time, while water content was influenced significantly by time of filleting and brining. Brining and storage time significantly influenced the whiteness, while time of filleting and storage time had significant influence upon the fillet index. The growth of Photobacterium phosphoreum was affected by storage time only, and the pH of the fillets was not influenced significantly by any of the parameters tested.     

Biochemical,physical and sensory quality of ice-stored Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) as affected by pre-slaughter stress,percussion stunning and AQUI-S<Superscript>™</Superscript> anaesthesia

Biochemical, physical and sensory quality of farmed Atlantic cod subjected to percussion stunning (control), anaesthesia (AQUI-S^™) and excessive exercise (30 min chasing before slaughter, ‘stressed’) were analysed after 7 days of ice storage. The white muscle energy status (initial pH, muscle twitches and high-energy phosphates) revealed that the fish were truly representatives of rested (percussion stunned and anaesthetised) and stressed cod. Sensory evaluation showed that the fillets of cod exposed to percussion stunning and AQUI-S^™ anaesthesia prior slaughter had slightly higher whiteness scores, and that the fillets of AQUI-S^™ anaesthetised cod had slightly shinier surfaces than the fillets of cod exposed to pre-slaughter stress. Furthermore, fillets of anaesthetised (AQUI-S^™) cod had significantly higher inosine monophosphate (IMP) contents and lower K-values than fillets of cod exposed to pre-slaughter stress, after 7 days of ice storage. Pre-slaughter stress did to some extent affect fillet colour immediately after killing and after ice storage. Otherwise, no significant effects of stress were observed with regard to biochemical, physical or sensory quality (ultimate pH, water content, drip loss, water holding capacity, texture and gaping) of farmed Atlantic cod.     

Purification and characterisation of cathepsin L from the skeletal muscle of blue scad (Decapterus maruadsi) and comparison of its role with myofibril-bound serine proteinase in the degradation of myofibrillar proteins

The modori phenomenon during surimi production is caused by endogenous proteinases, especially cathepsin L and myofibril-bound serine proteinase (MBSP). Cathepsin L from the skeletal muscle of blue scad (Decapterus maruadsi) was purified to homogeneity by ammonium sulphate fractionation and a series of column chromatographies and revealed a single band with molecular mass of 30 kDa on SDS–PAGE. Peptide mass fingerprinting (PMF) obtained three fragments with 48 amino acid residues, which were highly identical to cathepsin L from other fish species. Its optimal pH and temperature were 5.5 and 55 °C, respectively. Meanwhile, MBSP was purified from the skeletal muscle of blue scad, and the roles of cathepsin L and MBSP in the degradation of myofibrillar proteins were compared. The results indicated that MBSP is more effective than cathepsin L in promoting the degradation of myofibrillar proteins, especially myosin heavy chain (MHC), suggesting that MBSP plays a more significant role.     

The effect of postmortem processing treatments on quality attributes of raw Atlantic salmon (Salmo salar) measured by sensory and instrumental methods

The main objective of this study was to compare the effects of three different processing treatments on sensory attributes and instrumental quality measurements of raw Atlantic salmon (Salmo salar) fillets. Salmon was either pre-rigor filleted and restricted or allowed to contract (Vacuum-PRE and Contracted-PRE, respectively) or post-rigor filleted (POST). Sensory evaluation (appearance, flavour, texture) and instrumental quality measurements (colour, texture, fat, astaxanthin, liquid holding capacity) were performed at 5–7 days postmortem. Sensory evaluation revealed that Vacuum-PRE fillets had less desirable quality attributes than the other treatment groups, with higher scores for tenderness and whiteness and lower scores for hardness and colour intensity. The observed changes in fillet height between the treatment groups indicated that the immediate vacuum packaging of the Vacuum-PRE fillets had limited their contraction during rigor mortis development, resulting in the negative effects observed on quality. This implicates that the well-known positive effects of pre-rigor filleting regarding colour and texture can be reduced or even abolished if the fillets are restricted from contraction during rigor mortis development.     

Effects of fasting prior to slaughter on technological and sensory properties of the loin muscle (M. longissimus dorsi) of pigs

The aim of this study was to investigate the effect of pre-slaughter fasting time, sex and feeding regime on water-holding capacity (WHC), colour and sensory properties and their relationship with pH in M. longissimusdorsi (LD) in pigs. 270 pigs of the commercial Norwegian crossbreed Noroc (Norwegian Landrace × Yorkshire sow and Norwegian Landrace × Duroc boar) were used involving two sexes (gilts and castrates), two feeding regimes (restricted and ad libitum) and four fasting treatments: (F4) 4 h fasting (control), (F175) 17.5 h fasting on the farm, (FO175) 17.5 h fasting overnight at the abattoir, and (FO265) 26.5 h fasting overnight at the abattoir. Additionally, the pigs experienced two abattoir lairage times as fasting treatments F4 and F175 had a lairage time of 1.5 h, while fasting treatments FO175 and FO265 had a lairage time of 23.0 h. A short fasting time of 4 h led to a delayed decline in pH post-mortem and a lower ultimate pH (pHu) in the LD compared with a fasting time of 26.5 h which resulted in a rapid pH decline early post-mortem and a high pHu. Prolonged fasting reduced drip loss, resulted in a darker colour and tended to improve tenderness of the LD. Castrates showed lower drip loss, higher lightness and improved tenderness and juiciness compared with gilts, while ad libitum feeding improved tenderness compared to restricted feeding. There are obvious negative relationships between pHu and drip loss, lightness and tenderness of LD.     

Protease activity impacts flesh colour during post-mortem storage of farmed Atlantic halibut (Hippoglossus hippoglossus): A condition referred to as chalky halibut

In the present study, fillet colour, pH and the activities of cathepsin B, B + L, D, H, collagenase and calpain were measured in 50 Atlantic halibut (Hippoglossus hippoglossus) farmed and harvested at Mørkvedbukta research station (Bodø University College) in May, 2009. May is a period of the year when a portion of the fish can temporarily lose its translucent colour, becoming milky instead, a phenomena referred to as chalky halibut. The aim of the present study was to discover whether enzymes, known to be involved in flesh quality degradation, are also involved in the post-mortem discoloration of halibut flesh. Colour assessment was performed using a Minolta instrument (L value) and a linear regression showed that pH had the most pronounced effect on chalkiness (r² = 0.81, p < 0.001). Of the investigated enzymes, cathepsins B and D explained 35 and 13% of the total variation, respectively (p < 0.01). In addition, an unexpected inverse relationship between chalkiness and calpain was found (r² = 0.48, p < 0.001). The present results suggest that the activity of cathepsins contributes to the development of chalkiness in Atlantic halibut.     

Association between cathepsin L (CTSL) and cathepsin S (CTSS) polymorphisms and meat production and carcass traits in Italian Large White pigs

The aim of this work was to identify and analyse DNA markers in two cathepsin genes, cathepsin L (CTSL) and cathepsin S (CTSS), and study their association with meat quality parameters (including cathepsin activity of post mortem muscle) and several production traits in Italian Large White pigs. In silico analysis of the porcine CTSL and CTSS genes identified two new single nucleotide polymorphisms (SNPs), one located in exon 5 (CTSL) and one positioned in the 5′-untranslated region (CTSS). Allele frequencies of these two SNPs were evaluated in 7 different pig breeds. In addition, radiation hybrid mapping localized the CTSS gene on porcine chromosome 4, in an important QTL region for meat and fat deposition. Association analysis with meat quality traits, carried out in 268 Italian Large White pigs, indicated association of the CTSL polymorphism with average daily gain (ADG), weight of lean cuts (LC) and back fat thickness (BFT) estimated breeding values (P < 0.05). Results for LC and BFT were also confirmed using a selective genotyping approach in other Italian Large White pigs (P < 0.01). In the 268 Italian Large White pigs, the CTSS polymorphism was associated with feed:gain ratio and ADG (P < 0.05). No association was observed between the analysed markers and meat quality parameters (pH₁, pH_u, lactate, glycogen, glycolytic potential and cathepsin activity, all measured on Musculus semimembranosus). Overall these results, together with those previously reported for other genes of this family, suggest that cathepsin genes play a role in defining economically important traits in pigs.     

Sensory,physical and chemical characteristics of cooked ham manufactured from rapidly chilled and earlier deboned M. semimembranosus

Effects of rapid chilling of carcasses (at − 31 °C in the first 3 h of chilling, and then at 2–4 °C) and earlier deboning (8 h post-mortem), compared to rapid (till 24 h post-mortem) and conventional chilling (at 2–4 °C, till 24 h post-mortem), on quality characteristics of pork M. semimebranosus and cooked ham were investigated. Quality measurements included pH value, colour (CIEL*a*b* values) and total aerobic count of M. semimebranosus, as well as sensory (colour, juiciness, texture, and flavour), physical (pH value, colour – CIEL*a*b* values and texture – Warner–Bratzler shear and penetration forces) and chemical (protein, total fat, and moisture content) characteristics of cooked ham. The cooked ham was manufactured from pieces of M. semimebranosus with ultimate lightness (CIEL* value) lower than 50. Rapid chilling and earlier deboning significantly increased quantity of M. semimebranosus desirable for cooked ham manufacturing. Earlier start of pork fabrication did not affect important quality characteristics of cooked ham.     

Contribution of cathepsins B,L and D to muscle protein profiles correlated with texture in rainbow trout (Oncorhynchus mykiss)

Post-mortem softening of fish tissue often results in low yield and decreased product quality. In this study, proteolytic profiles of trout stored 5 days on ice were obtained by SDS–PAGE. The link between protein band intensities and firmness of trout fillets was examined through a correlation study. In parallel, trout extracts were incubated with cathepsin B, cathepsin L and cathepsin D, alone or in combination, in order to evaluate the effect of each cathepsin on the texture-related proteins. Proteins from both myofibrillar (α-actinin, actin, MLC1, MLC2, and N-terminal 70 kDa MHC fragment) and sarcoplasmic (glycogen phosphorylase, creatine kinase, and TPI) fractions correlated closely with firmness. Cathepsins D, B and L affected, respectively, 10, 9 and 4 out of the 17 protein bands correlating with firmness, and most changes induced by cathepsin D were unfavourable to firmness. This implies that cathepsin D is likely to be involved in textural change of trout, due to breakdown of the muscle structure.     

Low salt brining of pre-rigor filleted farmed cod (Gadus morhua L.) and the effects on different quality parameters

Pre-rigor processing of cod fillets may have economic benefits, but this potential has usually been overshadowed by process-linked difficulties such as pin bone removal, rapid rigor onset and higher drip losses. The aim of this work was to study the impact on fillet quality parameters after immersing pre-rigor filleted farmed cod in different NaCl solutions ranging from 15 to 60 g/L. Temperature of the fish at death was 4 °C, in immersion solutions 2 °C, and following immersion the fillets were stored in ice within plastic bags for 14 days. As controls, one group was filleted pre-rigor but not immersed, and one group was filleted post-rigor and not immersed. Immersing in salt solution resulted in better yield compared to both control groups. Higher salt content generally increased rigor contraction, but significantly reduced fillet gaping and the force required to pull pin bones. Thus, relatively low salt levels within the fillets had a positive impact on some of the problems associated with pre-rigor filleting.     

Physicochemical changes in ω − 3-enhanced farmed rainbow trout (Oncorhynchus mykiss) muscle during refrigerated storage

Physicochemical changes of ω − 3-enhanced farmed rainbow trout (Oncorhynchus mykiss) fillets developed by dietary modification with flaxseed oil and α-tocopheryl acetate (α-TA) were determined during storage at 2 °C. Trout were fed experimental diets for 120 days followed by processing to obtain boneless skinless fillets. The dietary modification increased concentration of total ω − 3 fatty acids in the fillets, which enhanced chances for lipid oxidation during storage. The fillets were vacuum or non-vacuum packed and stored at 2 °C for 10 or 12 days. Dietary α-TA resulted in higher (P < 0.05) concentration of α-tocopherol in fillets during storage; however, it did not retard (P > 0.05) lipid oxidation. Vacuum packaging resulted in much lower (P < 0.05) TBARS and higher (P < 0.05) retention of α-tocopherol during storage than non-vacuum packaging. However, α-tocopherol unlike vacuum packaging better protected ω − 3 FA in the fillets during storage.     

Early post-mortem sarcoplasmic proteome of porcine muscle related to protein oxidation

Oxidative deterioration or modifications of proteins which appear during meat storage and processes can result in the impairment of technological, sensorial and nutritional qualities. Improving the quality involves a better understanding of the biochemical mechanisms responsible for protein oxidation in meat. For that purpose, an analysis was conducted to investigate the relationships between the early post-mortem sarcoplasmic proteome, which contains the majority of enzymes involved in the oxidative process, and protein oxidation generated during meat storage and cooking. This study was performed in Longissimus lumborum pig muscle. In order to have sufficient variability in the proteome and in the meat oxidation level, five groups of 10 animals issued from two different breeds and raised in three different rearing systems were analysed. Protein oxidation was estimated by the measurement of carbonyl groups after 1 and 4 days of refrigerated storage, and after 100 °C experimental cooking of the 4 days aged meat. Significant correlations (p < 0.05) were observed between the level of carbonyl groups and the intensities of 104 spots of the 2D electrophoresis, out of which 52 were clearly identified. The possible involvement of some proteins in the muscle oxidative stress leading to protein oxidation is discussed.     

Effects of dietary lipid source on fillet chemical composition,flavour volatile compounds and sensory characteristics in the freshwater fish tench (Tinca tinca L.)

In this study the effects of soybean and linseed oils on chemical and sensory characteristics of fillets were evaluated in the freshwater fish tench (Tinca tinca L.). Five experimental diets, differing only in the relative amount of soybean and linseed oil, were formulated and the experiment was conducted on 360 sub-adult tench for 12 weeks. The fatty acid composition of muscle reflected that of the diets and significant correlations were observed. Diets containing higher amounts of n − 6 fatty acids were responsible for an increased level of n − 6 fatty acids in the fish flesh. Consequently, an increase in the relative amount of n − 6-derived volatile aldehydes was also observed. These latter compounds are generally reported to contribute negatively to the general aroma of fish muscle and, consistently, the results of the sensory analysis showed a high value for the “off-flavour” attribute for fish fed the diet containing only n − 6-rich soybean oil.