A Proof-of-Concept Study to Develop a Peptide-Based Vaccine against Salmon Lice Infestation in Atlantic Salmon (Salmo salar L.).

Proteins present in blood samples from Atlantic salmon (Salmo salar) infected with salmon lice (Lepeophtheirus salmonis) were analyzed using liquid chromatography-high-resolution mass spectrometry. Bioinformatic analyses revealed 1820 proteins, of which 58 were assigned to lice. Among these, peroxiredoxin-2, an antioxidant protein, was found relevant with respect to blood feeding of the parasite. The three-dimensional structure analysis of the protein revealed a surface amino acid sequence of interest. A 13-amino-acid peptide was selected as a potential antigen due to its predicted solubility, antigenicity, probable non-allergenic, and non-toxic nature. This peroxiredoxin-2-derived peptide was synthesized, combined with a commercially available adjuvant, and used for vaccination. The test vaccine demonstrated a 60-70% protection rate against early-stage Lepeophtheirus salmonis infection in a challenge trial in Norway. Additionally, the vaccine was tested against salmon lice (Caligus rogercresseyi) in Chile, where a remarkable 92% reduction in the number of adult lice was observed. Thus, in combination with the selected adjuvant, the peptide showed antigenic potential, making it a suitable candidate for future vaccine development. The approach described holds promise for the development of peptide vaccines against various ectoparasites feeding on blood or skin secretions of their hosts.

A comparative study of diploid versus triploid Atlantic salmon (Salmo salar L.). The effects of rearing temperatures (5, 10 and 15°C) on raw material characteristics and storage quality.

Several major market operators argue that the current level of knowledge about quality is too scant to justify a switch to a large-scale production of triploid salmon. The aim of the present study was, therefore, to elucidate how rearing conditions (5, 10 and 15°C) affect the flesh quality of triploid Atlantic salmon (Salmo salar L., 1.6±0.3kg). As a reference, diploid salmon kept under equal conditions and with equal genetics were used. The main design discriminant was the holding temperature; increased temperature gave increased blood lactate, rigor index (Ir), drip loss (DL), content of astaxanthin and intensity of redness, but reduced muscle pH, cathepsin activity and fillet lightness. Salmon kept at 10°C grew the fastest. It is concluded that ploidy gave less variation than temperature. Triploids were characterized by lower blood haematocrit (Hct) and Ir, higher DL and collagenase activity, and on average, paler and less yellowish fillets.

Determination of the myoglobin states in ground beef using non-invasive reflectance spectrometry and multivariate regression analysis.

Seventy-two samples of ground beef from M. semimembranosus of two 5 and two 1.5year old animals were prepared. Two types of fat tissues from either beef or pork were added to the ground beef. The samples were prepared to contain predominantly deoxymyoglobin (DMb), oxymyoglobin (OMb) and metmyoglobin (MMb) states on surfaces using selected methods based on chemical treatment (for MMb) and oxygen pressure packaging to induce the two other states. Reflectance spectra were measured on ground beef after three storage times. Partial least regression analysis was used to make calibration models of the desired myoglobin states. Validated models using leave-one-sample out cross validation gave, after correction and normalization, prediction errors of about 5%. Long term storage of ground beef was unsuitable for preparing pure MMb states due to gradual reduction of the pigment to DMb, presumably by bacteria.

Slaughter of Atlantic salmon (Salmo salar L.) in the presence of carbon monoxide.

The different stunning methods for Atlantic salmon can still be improved with regard to animal welfare. Salmon exposed to carbon monoxide expressed no aversive reactions towards CO as such. CO exposed fish showed an earlier onset of rigour mortis and a faster decrease in muscle pH due to depletion of oxygen during the treatment. Exposure to CO did increase the level of cortisol compared to undisturbed control fish, but the increase was less than in the water only control group. Neuroglobin, a CO binding globin, was found in salmon brain and Saccus vasculosus, a richly vascularized sac connected to the fish brain. Binding of CO to neuroglobin during sedation might possibly improve animal welfare.

Cloning, expression and purification of Atlantic salmon (Salmo salar, L.) neuroglobin.

Neuroglobin (Ngb) exists only in small amounts in salmon brain. In order to study the protein in more detail salmon neuroglobin (sNgb) was cloned, heterologously expressed in Escherichia coli and purified. The protein had red color and showed the characteristic peaks at 411nm (metNgb), 415nm (carboxyNgb) and 424nm (deoxyNgb). Western analysis showed that sNgb reacted weakly against a rabbit anti human neuroglobin (hNgb) and strongly to a sNgb specific antibody. Our 3D-homology model of the sNgb indicated modifications adjacent to and in the O(2)/CO binding site. This may correlate to differences in substrate affinities for the sNgb compared to the hNgb. Also sNgb contained shorter helixes and longer interhelical loops typical for psychrophilic proteins.

New procedure for improving precision and accuracy of instrumental color measurements of beef.

The surface layers of steaks from bovine M. semimembranosus were prepared to have deoxy- (DMb), oxy- (OMb) and metmyoglobin (MMb) states using either chemicals (CHEM) or oxygen partial pressure packaging (OPP). Ninety-six different meat surface areas were measured in reflectance mode (400-1100 nm) for each preparation method. Reflectance spectra were converted to absorbance (A) and then transformed by Kubelka-Munk transformation (K/S) and/or extended multiplicative scatter correction (EMSC). Transformed spectra of prepared pure states were used to make calibration models of MMb, DMb and OMb using either selected wavelengths (SW) or partial least square (PLS) regression. Finally, the predicted myoglobin states were normalized to ensure that no state was <0 or >1 and the sum of all states equal to 1. Multivariate calibrations (i.e. PLS) outperformed the univariate calibrations (i.e. SW). The OPP method of preparing pure states was clearly best for OMb while the CHEM method was best for preparing MMb on fresh meat surfaces. Both preparation methods needed improvement concerning DMb. The CHEM(K/S) SW and the OPP EMSC(A) PLS methods predicted MMb, DMb and OMb with root-mean-square errors of cross validation (RMSECV) equal to 0.08, 0.16 and 0.18 (range 0-1) and 0.04, 0.04 and 0.04 (range 0-1), respectively. This new reflectance protocol has potential for routine meat color measurements.

Composition and some quality characteristics of the longissimus muscle of reindeer in Norway compared to farmed New Zealand red deer.

Norwegian reindeer of Finnmark county live under harsh conditions on extensive feed sources. Thus the meat may have special qualities. M. longissimus lumborum from 30 animals was investigated with respect to carcass and meat quality and compositional/nutritional characteristics. Meat from calves had a higher myofibrillar fragmentation index and tenderness, and had lighter, redder and more yellow colour than meat from adult reindeer. Regarding nutritional compounds the meat from calves had lower antioxidant capacity, and higher taurine levels than adults, while the levels of iron, carnosine, anserine, and vitamin E were similar. Carcass weights of the adult reindeer were lower than the weights reported for Swedish reindeer and New Zealand farmed red deer. Reindeer muscles had higher antioxidant capacity, shorter sarcomeres, smaller muscle fibre diameters, higher n-6/n-3 ratios of fatty acids in the intramuscular fat and higher muscle taurine levels relative to values for the same muscle of New Zealand farmed red deer.

Oxygen consumption rate of permeabilized cells and isolated mitochondria from pork M. masseter and liver examined fresh and after freeze-thawing at different pH values.

UNLABELLED: The oxygen consumption rate (OCR) of 2 types of permeabilized tissues and their corresponding isolated mitochondria from porcine M. masseter and liver, resulting in 4 systems, was studied at different pH values (5.0 to 7.1) using fresh samples and samples frozen directly in liquid nitrogen (N2) or air-frozen at -20°C. A protocol with the additive sequence rotenone-succinate-ADP (adenosine diphosphate)-cytochrome c-FCCP (carbonyl cyanide p-trifluoromethoxyphenylhydrazone) was used to study respiration changes. The OCR of liver respiring on succinate (OCR(S)) was higher than that of muscle tissue. pH had a larger effect on OCR(S) than freeze-thawing. Low pH was associated with reduced OCR(S). The OCR(S) of isolated muscle mitochondria appeared to be an underestimated relative to the OCR(S) of permeabilized muscle cells. Increasing pH, following prior subjection to pH 5.0, showed partial reversibility of the OCR(S). The freeze-thaw cycle increased the OCR(S) when muscle systems were frozen and examined above pH 6.0; this effect was less apparent for liver tissue. A response to cytochrome c addition, indicating a defective outer mitochondrial membrane, was observed for all 4 systems. The response was, however, lowest for permeabilized cells. The ADP/FCCP additive pair indicated partial coupling for isolated liver and muscle mitochondria. These additives gave weak responses for the permeabilized liver cells while the OCR seemed to be inhibited for permeabilized muscle fibers when ADP/FCCP was added. PRACTICAL APPLICATION: The mitochondrial state is believed to be important for myoglobin reduction, development of flavor, and possibly other meat qualities. By monitoring the oxygen consumption in mitochondria and meat we can better understand and control such processes following freezing and thawing.

Effect of various electric field strengths and current durations on stunning and spinal injuries of Atlantic herring.

The aim of this study was to evaluate the effect of electric field strength and current duration on wild-caught Atlantic herring Clupea harengus stunned with sinusoidal 50-Hz AC in seawater. The fish were exposed to electric field strengths ranging from 16 to 142 V/m and current durations from 1 to 12 s. We recorded the elapsed times between the point at which each fish became unconscious and the points at which it resumed normal behavioral functions. We also investigated injuries such as broken spinal columns and hematomas after the fish were filleted. The threshold electric field strength required to stun all of the fish to unconsciousness was 33 V/m for 1 s. The duration of the unconscious condition increased as both electric field strength and current duration increased. Of a total of 260 Atlantic herring, 60% had broken spinal columns. The proportion of fish with fractured spines was independent of field strength and duration, but the number of fractures per fish increased with field strength. We conclude that electrical stunning would promote the welfare of Atlantic herring that are landed alive but negatively affect fillet quality owing to hematomas associated with the fractures.