Variation among stallions in sperm quality after single layer centrifugation

Although single layer centrifugation (SLC) selects robust spermatozoa from stallion semen, the effect of individual variation has not been studied in detail. The objective of this study was to determine the variation among stallions in the effects of SLC on sperm quality during cooled storage for up to 48 hr. Semen samples from seven stallions (18 ejaculates) were split, with one portion being used for SLC and the other serving as a control (CON). Sperm quality (kinematics, reactive oxygen species (ROS) production, membrane integrity (MI) and chromatin integrity) were analysed at 0, 24 and 48 hr using computer-assisted sperm analysis and flow cytometry. Sperm quality was better in SLC than in CON at all timepoints, especially chromatin integrity and MI (p < .0001 for both), and some categories of ROS production (e.g. proportion of live hydrogen peroxide negative spermatozoa, p < .0001), but the degree of improvement varied among stallions and type of ROS (p < .05–p < .0001). Total and progressive motility were also better in SLC samples than in CON at 24 and 48 hr (p < .0001), although the effect on sperm kinematics varied. The interaction of treatment, time and stallion was not significant. In conclusion, sperm quality was better in SLC samples than in CON, although there was considerable individual variation among stallions. The improvement in sperm quality, particularly in chromatin integrity, was clearly beneficial, and therefore the use of this technique would be warranted for all stallion semen samples.     

Distribution and induction of cytochrome P450 1A1 in the rainbow trout brain

Cytochrome P450 (CYP) 1A1 participates in the activation as well as detoxification of environmental pollutants such as aromatic hydrocarbons. This CYP form is also efficiently induced by aromatic hydrocarbons. The presence of CYP 1A1 in the brain might thus be of physiological and toxicological importance. In the present investigation on rainbow trout, the distribution of 7-ethoxyresorufin-O-deethylase (EROD) activity, a cytochrome CYP 1A1 catalyzed reaction, was measured in whole tissue homogenates from brain parts. In control fish, a relatively high activity was found in the rainbow trout olfactory bulb compared to the other brain parts. Although an EROD induction (3 to 7-fold) by β-naphthoflavone (BNF) was recorded in all brain parts from the rainbow trout, the highest induced activity was measured in the olfactory bulbs. To ascertain the distribution of EROD activity in cells, whole brain tissue was subfractionated by differential centrifugation. The fractionation scheme separated mitochondria (P2 fraction) and microsomes (P3 fraction) as determined by marker enzymes and electron microscopy. In control rainbow trout, a low EROD activity could be measured in the P2 fraction. BNF induced the EROD activity in both P2 and P3 fractions. Western blotting showed the induction by BNF of a protein band in the P2 and P3 fractions with a molecular mass around 58,000 when highly specific anti-cod CYP 1A1 antibodies were used. ELISA measurements confirmed the induction of CYP 1A1 protein in the rainbow trout brain subcellular fractions.     

Automated image analysis as a tool to quantify the colour and composition of rainbow trout (Oncorhynchus mykiss W.) cutlets

The goal of this paper is to propose and evaluate automated image analysis methods for describing muscle cutlets in rainbow trout. The proposed automated image analysis methods were tested on a total of 983 scanned images of trout cutlets, and included quality traits such as fat percentage, flesh colour and the size of morphologically distinguishable subparts of the cutlet. A sub-sample of 50 images was randomly selected for manual segmentation of the cutlet, the dorsal fat depot and the red muscle and regions. The identification of these regions by manual and automatic image analysis correlated strongly (r = 0.97, r = 0.95 and r = 0.91, respectively). The estimated fat percentage obtained from image analysis, based on the area of visible fat and the colour of the cutlet flesh, correlated well with chemical fat percentage measured by mid-infrared transmission spectroscopy (MIT) (r = 0.78). The automated image analysis methods are therefore a reliable means of predicting the fat percentage of trout cutlets. Principal component analysis (PCA) loading plots were used to identify subsets of variables from the image analysis of special significance for further studies; cutlet area, dorsal fat depot area, red muscle area, back height, cutlet width, and width of left and right abdomen wall were among the variables selected. PCA loading plots of different colour variables indicated that simple statistical coefficients such as percentiles and mean values can be used to quantify different aspects of flesh colour. In conclusion, the methods presented here provide a powerful toolbox for describing important morphological structures and quality traits of trout cutlets.     

Comparative evaluation of the emulsifying properties of phosphatidylcholine after enzymatic acyl modification

The ability of enzymatically synthesized structured phosphatidylcholine (PC) containing caprylic acid to form and stabilize oil-in-water emulsions prepared with different triglycerides [medium chain triglycerides (MCT), soybean oil, and enzymatically synthesized structured lipids] was examined and compared with natural soybean PC and deoiled lecithin. Emulsions were prepared with varying oil and emulsifier concentrations. The particle size distribution, creaming stability, and viscosity were measured for the evaluation of the emulsifying properties. With an increase in the oil concentration, there was an increase in particle size, viscosity, and creaming layer. With an increase in the phospholipid (PL) concentration, there was usually a decrease in particle size and an increase in viscosity, where the emulsion stability was increased. General emulsions prepared with structured lipids resulted in smaller particle sizes as compared to MCT and soybean oil. Deoiled lecithin was able to increase the viscosity more significantly and give smaller particle sizes as compared to the other emulsifiers, thus producing more stable emulsions. However, in certain cases, structured PC was superior to deoiled lecithin and soybean PC. This observation was made for emulsions prepared with soybean oil or structured lipid at an oil/water ratio of 10:90. At an oil/water ratio of 30:70, the deoiled lecithin performed better as compared to the other PLs with all oil types. However, structured PC produced more stable emulsions as compared to natural soybean PC in MCT and soybean oil.     

Customer adapted grading of Scots pine sawn timber using a multivariate method

To define new grading rules, or to customize the ones in use in a rule-based automatic grading (RBAG) system of boards, is a time-consuming job for a sawmill engineer. This has the effect that changes are rarely made. The objective of this study was to continue the development of a method that replaces the calibration of grading rule settings by a holistic-subjective automatic grading, using multivariate models. The objective was also to investigate if this approach can improve sawmill profitability and at the same time have a satisfied customer. For the study, 323 Scots pine (Pinus sylvestris L.) boards were manually graded according to the preferences of an important customer. That is, a customer that regularly purchases significant volumes of sawn timber. This manual grading was seen as reference grading in this work. The same boards were also scanned and graded by a RBAG system, calibrated for the same customer. Multivariate models for prediction of board grade based on aggregated knot variables, obtained from the scanning, were calibrated using partial least squares regression. The results show that prediction of board grades by the multivariate models were more correct, with respect to the manual grading, than the grading by the RBAG system. The prediction of board grades based on multivariate models resulted in 76–87% of the boards graded correctly, according to the manual grading, while the corresponding number was 63% for the RBAG system.