Biofluorescent response in lumpfish Cyclopterus lumpus to a therapeutic stressor as assessed by hyperspectral imaging.

The demand for lumpfish (Cyclopterus lumpus) as a biological control for salmon lice is increasing. However, lumpfish welfare is considered a limiting factor within aquaculture operations. Identifying a noninvasive parameter that measures subclinical stress in lumpfish is a key goal for improving their welfare. The lumpfish is documented to emit green and red biofluorescence within the blue shifted light of their environment. Here we show that lumpfish fluorescence responds to a therapeutic stressor within a controlled experiment. Lumpfish (n = 60) underwent a 3-h freshwater bath therapeutant to evaluate whether fluorescence spectra produced by the species respond to external stimuli. Lumpfish were quickly scanned under a hyperspectral camera (400-1000 nm spectral range) prior to and after treatment. The lumpfish were randomly divided into 3 groups with identical treatment. All groups increased fluorescence emissions, though the level of change depended on whether the averaged, red, or green spectra were analyzed; the control group (n = 20) remained constant. All lumpfish emitted green fluorescence (~ 590-670 nm) while a portion (49%) produced red fluorescence (~ 690-800 nm). As lumpfish fluorescence shifts in response to the applied stressor, this study provides insight into how fluorescence may be incorporated into the welfare management of lumpfish.

Hidden in plain sight: hyperspectral documentation of complex biofluorescence produced by the green sea urchin (Strongylocentrotus droebachiensis).

Biofluorescence in echinoderms is largely unexplored, and even though the green sea urchinStrongylocentrotus droebachiensisis a well-studied species, the presence and/or function of fluorescence remains very poorly understood. Hyperspectral imaging was conducted on adult sea urchins (N = 380) while fluorospectrometric analysis was conducted on sea urchin coelomic fluid (N = 30). Fluorescence was documented in both the spines and coelomic fluid ofS. droebachiensis. Intact spines exhibited a low intensity green emission (∼550-600 nm), while broken spines averaged a high emission peak in the green spectrum (∼580 nm). Sea urchins produce a red exudate with a pronounced emission peak (∼680 nm) with a shoulder peak (∼730 nm). The sampled coelomic fluid exhibited high variability, with a majority exhibiting a low-level green fluorescence while pronounced emission peaks (N = 5) were found in the red spectrum (∼680 nm). The complex fluorescence produced byS. droebachiensiswarrants further investigation on its applicability for monitoring welfare of sea urchins in aquaculture facilities.

Raman spectroscopy and NIR hyperspectral imaging for in-line estimation of fatty acid features in salmon fillets.

Raman spectroscopy was compared with near infrared (NIR) hyperspectral imaging for determination of fat composition (%EPA + DHA) in salmon fillets at short exposure times. Fillets were measured in movement for both methods. Salmon were acquired from several different farming locations in Norway with different feeding regimes, representing a realistic variation of salmon in the market. For Raman, we investigated three manual scanning strategies; i) line scan of loin, ii) line scan of belly and iii) sinusoidal scan of belly at exposure times of 2s and 4s. NIR images were acquired while the fillets moved on a conveyor belt at 40 cm/s, which corresponds to an acquisition time of 1s for a 40 cm long fillet. For NIR images, three different regions of interest (ROI) were investigated including the i) whole fillet, ii) belly segment, and iii) loin segment. For both Raman and NIR measurements, we investigated an untrimmed and trimmed version of the fillets, both relevant for industrial in-line evaluation. For the trimmed fillets, a fat rich deposition layer in the belly was removed. The %EPA + DHA models were validated by cross validation (N = 51) and using an independent test set (N = 20) which was acquired in a different season. Both Raman and NIR showed promising results and high performances in the cross validation, with R2CV = 0.96 for Raman at 2s exposure and R2CV = 0.97 for NIR. High performances were obtained also for the test set, but while Raman had low and stable biases for the test set, the biases were high and varied for the NIR measurements. Analysis of variance on the squared test set residuals showed that performance for Raman measurements were significantly higher than NIR at 1% significance level (p = 0.000013) when slope-and-bias errors were not corrected, but not significant when residuals were slope-and-bias corrected (p = 0.28). This indicated that NIR was more sensitive to matrix effects. For Raman, signal-to-noise ratio was the main limitation and there were indications that Raman was close to a critical sample exposure time at the 2s signal accumulation.

Spectroscopic Techniques for Monitoring Thermal Treatments in Fish and Other Seafood: A Review of Recent Developments and Applications.

Cooking is an important processing method, that has been used since ancient times in order to both ensure microbiological safety and give desired organoleptic properties to the cooked food. Fish and other seafood products are highly sensitive to thermal treatments and the application of severe heat can result in negative consequences on sensory and nutritional parameters, as well as other quality attributes of the thermally processed products. To avoid such undesired effects and to extend the shelf life of these perishable products, both the heat processing methods and the assessment techniques used to monitor the process should be optimized. In this review paper, the most common cooking methods and some innovative ones will first be presented with a brief discussion of their impact on seafood quality. The main methods used for monitoring heat treatments will then be reviewed with a special focus on spectroscopic techniques, which are known to be rapid and non-destructive methods compared to traditional approaches. Finally, viewpoints of the current challenges will be discussed and possible directions for future applications and research will be suggested. The literature presented in this review clearly demonstrates the potential of spectroscopic techniques, coupled with chemometric tools, for online monitoring of heat-induced changes resulting from the application of thermal treatments of seafood. The use of fluorescence hyperspectral imaging is especially promising, as the technique combines the merits of both fluorescence spectroscopy (high sensitivity and selectivity) and hyperspectral imaging (spatial dimension). With further research and investigation, the few current limitations of monitoring thermal treatments by spectroscopy can be addressed, thus enabling the use of spectroscopic techniques as a routine tool in the seafood industry.

Use of Spectroscopic Techniques for a Rapid and Non-Destructive Monitoring of Thermal Treatments and Storage Time of Sous-Vide Cooked Cod Fillets.

In this work, the potential of spectroscopic techniques was studied to investigate heat-induced changes occurring during the application of thermal treatments on cod (Gadus morhua L.) fillets. Vacuum-packed samples were thermally treated in a water bath at 50, 60, 70 and 80 °C for 5 and 10 min, and further stored for one, four, and eight days at 4 ± 1 °C before analysis. Several traditional (including cooking loss, drip loss, texture, protein solubility, protein oxidation, and color) and spectroscopic (fluorescence and diffuse reflectance hyperspectral imaging) measurements were conducted on the same samples. The results showed a decrease in fluorescence intensity with increasing cooking temperature and storage time, while the impact of cooking time was only noticeable at low temperatures. Diffuse reflectance data exhibited a decrease in absorbance, possibly as a result of protein denaturation and increased scattering at higher cooking temperatures. Both fluorescence and diffuse reflectance data were highly correlated with color parameters, whereas moderate correlations were observed with most other traditional parameters. Support vector machine models performed better than partial least square ones for both classification of cod samples cooked at different temperatures and in prediction of the cooking temperature. The best classification result was obtained on fluorescence data, achieving an accuracy of 92.5%, while the prediction models resulted in a root mean square error of prediction of cooking temperature lower than 5 °C. Overall, the classification and prediction models showed good results, indicating that spectroscopic techniques, especially fluorescence hyperspectral imaging, have a high potential for monitoring thermal treatments in cod fillets.

Performance of Fluorescence and Diffuse Reflectance Hyperspectral Imaging for Characterization of Lutefisk: A Traditional Norwegian Fish Dish.

Lutefisk is a traditional Norwegian fish dish made from dried fish, such as cod or other whitefish. In Norway and other Nordic countries, lutefisk is considered among the most popular dishes served during Christmas or other festive occasions. However, to date, little attention has been paid to this product, and available research on the quality, processing, and chemistry of lutefisk is still limited. The quality of this very delicate product, with a high pH value, depends on many factors, such as the initial quality of raw materials (stockfish), the quantity of lye used during the preparation process of lutefisk, and time during soaking in the lye and water, among others, making it challenging to both optimize processing and monitor the quality of lutefisk. In this study, four commercially available lutefisk brands (labelled as A, B, C, and D) were characterized using two online spectroscopic techniques, namely fluorescence and diffuse reflectance hyperspectral imaging, implemented on conveyor belts to mimic industrial applications. The samples were also analyzed by the use of an offline laboratory instrument based on visible/near infrared diffuse reflectance spectroscopy. Three traditional measurements, including texture, water content, and pH, were also conducted on the same samples. Supervised classification PLS-DA models were built with each dataset and relationships between the spectroscopic measurements and the traditional data were investigated using canonical correlations. The spectroscopic methods, especially fluorescence spectroscopy, demonstrated high performance for the discrimination between samples of the different brands, with high correlations between the spectral and traditional measurements. Although more validations of the results of this study are still required, these preliminary findings suggest that the destructive, laborious, and time-consuming traditional techniques can be replaced by rapid and nondestructive online measurements based on hyperspectral imaging used in fluorescence or diffuse reflectance mode.