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.

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.

Simulated trawling: Exhaustive swimming followed by extreme crowding as contributing reasons to variable fillet quality in trawl-caught Atlantic cod (Gadus morhua).

Trawl-caught Atlantic cod (Gadus morhua) often yield highly variable fillet quality that may be related to capture stress. To investigate mechanisms involved in causing variable quality, commercial-sized (3.5±0.9 kg) Atlantic cod were swum to exhaustion in a large swim tunnel and subsequently exposed to extreme crowding (736±50 kg m-3) for 0, 1 or 3 hours in an experimental cod-end. The fish were then recuperated for 0, 3 or 6 hours in a net pen prior to slaughter to assess the possibility to reverse the reduced fillet quality. We found that exhaustive swimming and crowding were associated with increased metabolic stress, as indicated by increased plasma cortisol, blood lactate and blood haematocrit levels, accompanied by reduced quality of the fillets due to increased visual redness and lower initial muscle pH. The observed negative effects of exhaustive swimming and crowding were only to a small degree reversed within 6 hours of recuperation. The results from this study suggest that exhaustive swimming followed by extreme crowding can reduce fillet quality, as measured by fillet redness and muscle pH, and contribute to the variable fillet quality seen in trawl-caught Atlantic cod. Recuperation for more than six hours may be required to reverse these effects.

The absolute sensitivity of digital colour cameras.

A new and improved method to obtain the average spectral pixel responsivity and the quantum efficiency of Digital Single Lens Reflex (DSLR) cameras is outlined. Two semi-professional cameras, the Nikon D300 and the Canon 40D, are evaluated. The cameras red, green and blue pixel responsivities and quantum efficiency are retrieved by illuminating an integrating sphere with a wavelength tunable monochromator. 31 intensity calibrated monochromatic spectral lines from 4000 to 7000 A, with a bandpass of approximately 12 A, were used as a library to solve the main equations of observation for the cameras. Both cameras have peak sensitivity in the blue and minimum sensitivity in the red. The Canon 40D has blue and green channel sensitivity close to the Nikon D300. The Canon red channel has half the sensitivity of the Nikon camera.

Sensitivity calibration of digital colour cameras for auroral imaging.

A method to sensitivity calibrate Digital Single Lens Reflective (DSLR) cameras is outlined. A low intensity calibrated light source tunable in wavelength is described. 31 monochromatic lines from 4000 to 7000 A with a bandpass of approximately 12 A were used to find the spectral responses for the D70 and the D200 cameras manufactured by Nikon. The source radiance ranged from about 300 to 1.6k R/A. The cameras were operated in manual mode with 4 seconds exposure time at ISO 1600, which are typical settings required for night time photography of the aurora. For the Nikon D200 camera, the blue, green and red spectral responsivities peak at 4600, 5300 and 5900 A, respectively. The response was high for the blue colour channel with a clear cut-off at 4100 A for the UV part of the spectrum. The red channel response indicates low sensitivity above 6600 A. The D70 shows similar spectral responsivity, except that it peaks in the green colour channel and it is more sensitive to both UV and NIR radiation. Both cameras are capable of detecting night- and dayside auroral at 4 second exposure time. For optimal auroral imaging capability, the green and red spectral responsivities need to be shifted up by about 300 and 400 A in wavelength.

Effects of single wavelength selection for Anisakid roundworm larvae detection through multispectral imaging.

The occurrence of parasites in fillets of commercially important fish species affects both food quality and safety. Presently, the detection and removal of nematode parasites is done by inspection on a light table (candling) and manual trimming of the fillets. This operation is costly and time-consuming and is not effective for detecting and removing all the nematodes in the fillets. In the last decades, several alternative methods have been proposed, but these methods have failed to replace the candling method. A newly described method called imaging spectroscopy has produced promising results because the operator can record both spectral and spatial information from an object. In this work, we studied single-wavelength bands from a spectral image. Discrimination between nematodes and other objects in the fillets is dependent on the level of contrast. Quantification of the contrast in such images revealed that the level of contrast varied when different wavelengths were selected, and these variations are correlated with the absorption properties of the nematode. Visible light scatters greatly in fish muscle, generally complicating the detection of nematodes. In this study, light scattering was used in a way that reduces the background complexity in spectral images. When light scattering properties were used in a wavelength range different from the bulk of the nematode light absorption, spectral images with significantly higher contrast were produced.

Weakening Pin Bone Attachment in Fish Fillets Using High-Intensity Focused Ultrasound.

High Intensity Focused Ultrasound (HIFU) can be used for the localized heating of biological tissue through the conversion of sound waves into heat. Although originally developed for human medicine, HIFU may also be used to weaken the attachment of pin bones in fish fillets to enable easier removal of such bones. This was shown in the present study, where a series of experiments were performed on HIFU phantoms and fillets of cod and salmon. In thin objects such as fish fillets, the heat is mainly dissipated at the surfaces. However, bones inside the fillet absorb ultrasound energy more efficiently than the surrounding tissue, resulting in a "self-focusing" heating of the bones. Salmon skin was found to effectively block the ultrasound, resulting in a significantly lower heating effect in fillets with skin. Cod skin partly blocked the ultrasound, but only to a small degree, enabling HIFU treatment through the skin. The treatment of fillets to reduce the pin bone attachment yielded an average reduction in the required pulling force by 50% in cod fillets with skin, with little muscle denaturation, and 72% in skinned fillets, with significant muscle denaturation. Salmon fillets were treated from the muscle side of the fillet to circumvent the need for penetration through skin. The treatment resulted in a 30% reduction in the peak pulling force and 10% reduction in the total pulling work, with a slight denaturation of the fillet surface.

Compounds of parasitic roundworm absorbing in the visible region: target molecules for detection of roundworm in Atlantic cod.

The objective of this study was to contribute to the development of technology that will be able to replace manual operations in processing of fish fillets. Removal of parasites, black lining, remnants of skin, and bloodstains are costly and time-consuming operations to the fish processing industry. The presence of parasites in fish products tends to spoil consumers' appetites. Recent reports questioning the safety of eating cod infected with parasites might lower consumer acceptance of seafood. Presently, parasites are detected and removed manually. An average efficiency of about 75% under commercial conditions has been reported. In this study, we focused on biochemical differences between cod muscle and the prevalent anisakine nematode species (Anisakis simplex and Pseudoterranova decipiens) infecting Atlantic cod (Gadus morhua). Using reversed phase high-performance liquid chromatography equipped with a photodiode array detector, substances absorbing in the range 300 to 600 nm were identified in extracts from parasite material. These substances were not detected in extracts from cod tissue. Significant biochemical differences between cod muscle and parasite material have thus been demonstrated.

Spectral changes of Atlantic salmon ( Salmo salar L.) muscle during cold storage as affected by the oxidation state of heme.

The spectra of fresh salmon fillets change due to storage and packaging atmospheres. The aim of this study was to demonstrate the effects of heme oxidation states on spectral development in salmon fillets and to investigate the origin of a shoulder peak representing important spectral variations during storage. Hyperspectral images of fresh salmon fillets and mince with various water contents were collected during storage under different atmospheres. In addition, the absorption spectra of extracted salmon hemoglobin and its derivatives (methemoglobin and deoxyhemoglobin) were obtained. Air storage resulted in an increased similarity between spectra of methemoglobin and salmon fillets in principal component analysis. Results from the mince storage demonstrated that absorption features at the shoulder peak could be related to water content in the salmon muscle. This study established that the formation of oxidized heme is the primary source of spectral variations that occur during air storage of fresh salmon. Changes in the status of heme due to storage and packaging can influence the appearance of the underlying water absorption at the shoulder peak and create variations in the salmon spectra.

Visible/near-infrared spectroscopy detects autolytic changes during storage of Atlantic salmon (Salmo salar L.).

UNLABELLED: Visible (VIS)/near-infrared (NIR) spectroscopy was used to investigate spectroscopic changes occurring during storage of Atlantic salmon fillets with and without bacterial growth. A storage experiment was conducted for 11 d postmortem. Bacterial growth was inhibited by soaking a group of salmon fillets in 3 mM NaN3 prior to storage, while a control group retained its normal bacterial growth. Spectra were obtained by directly applying the spectroscopic probe onto the loin part of each fillet stored under conditions accelerating degradation. Principal component analysis (PCA) was used to monitor and compare spectroscopic development of the 2 groups and the results showed that VIS/NIR spectral changes occurred in the control as well as the treated group of samples within a single day after filleting. After 2 d of storage, stored samples were distinguishable from those fresh in both groups and it was only after the microbial spoilage became pronounced (8 to 9 log colony forming unit [CFU]/g) that the spectra of the spoiled control samples could be differentiated from spectra of the treated samples with no bacterial growth. Microbial growth is therefore not the only explanation for the spectral variations prior to microbial spoilage. Nonmicrobial, autolytic changes including possible changes in the physical properties are also contributing. Our results show that VIS/NIR spectroscopy can detect autolytic changes occurring in salmon muscle during the early stage of storage, independent of microbial growth. PRACTICAL APPLICATION: Important spectroscopic changes occur even when microbial growth is not apparent. This indicates that VIS/NIR spectroscopy may be used to determine the degree of freshness before microbial spoilage becomes relevant.

Automatic nematode detection in cod fillets (Gadus morhua) by transillumination hyperspectral imaging.

Traditional quality control of cod fillets is currently made by manual inspection on candling tables. This is a time consuming and expensive operation, contributing to a significant share of the cost with cod fillet production. In this study, transillumination hyperspectral imaging was implemented as a method for automatic nematode detection in cod fillets moving on a conveyer belt, and evaluated on industrially processed cod fillets. An overall detection rate of 58% of all nematodes (N= 922), with detection rate of 71% and 46% for dark and pale nematodes, respectively, is reported. This is comparable, or better, than what is reported for manual inspection under industrial conditions. The false alarm rate was high, with 60% of the fillets reported with one or more false alarms. These results show that the method is promising, but needs further refinements to reduce the false alarm rate and increase the imaging speed from 25 to 400 mm/s. Practical Application: Manual inspection of cod fillets is a huge bottleneck for the industry, accounting for half the production cost with cod fillet processing and reducing the processing speed. Transillumination hyperspectral imaging has the potential to reduce the manual labor required for cod fillet inspection and hence reduce the cost and increase the end product quality.

Effects of storage atmosphere and heme state on the color and visible reflectance spectra of salmon ( Salmo salar ) fillets.

It has previously been observed that the color of mackerel muscle is dependent on the status of heme as myoglobin and hemoglobin and hence the storage atmosphere. This study gives strong indications of this being the case also in salmon. Three different storage conditions were used to promote the oxidized, reduced, and carbon monoxide (CO) bound forms of heme in salmon and mackerel fillets. Color determination (instrumental color analysis, imaging, and sensory evaluation) and spectroscopic measurements were performed to study how spectral changes corresponded to color variations. Storage in CO significantly increased the redness in mackerel. This was also seen in salmon to such a degree that it was visible over normal levels of salmon carotenoids. Air storage increased the yellowness and reduced the redness in mackerel, but this effect was partly concealed in salmon by the astaxanthin absorption. The spectral differences due to storage condition could be ascribed to the spectral features characterizing heme of different oxidation states and bound to different ligands. The status of heme should therefore always be considered when experiments related to salmon color are performed. The findings could help in the understanding, control, and prediction of color loss in salmon during processing, storage, and transport.

Detection of nematodes in cod (Gadus morhua) fillets by imaging spectroscopy.

A promising method for detection of parasites in whitefish fillets has been developed. By use of imaging spectroscopy it is possible to record both spectral and spatial information from an object. In this work it is shown that by applying a white light transmission setup and imaging spectroscopy to cod (Gadus morhua) fillets, it is possible to make spectral images containing information to differentiate between fish muscle and parasites. The spectral images are analyzed by discriminant partial least square regression as well as image-filtering techniques. The method identifies parasites on the surface of the fillets as well as embedded parasites. One parasite was detected at 0.8 cm below the fillet surface, which is 2 to 3 mm deeper than what can be found by manual inspection of fish fillets. The method is nonintrusive and should thus be feasible for industrial purposes.