Development of hepatic steatosis in male and female mule ducks after respective force-feeding programs.

Male and female mule ducks were subjected to a force-feeding diet to induce liver steatosis as it is generally done only with male ducks for the production of foie gras. The different biochemical measurements indicated that the course of hepatic steatosis development was present in both sexes and associated with a huge increase in liver weight mainly due to the synthesis and accumulation of lipids in hepatocytes. In livers of male and female ducks, this lipid accumulation was associated with oxidative stress and hypoxia. However, certain specific modifications (kinetics of lipid droplet development and hepatic inflammation) indicate that female ducks may tolerate force-feeding less well, at least at the hepatic level. This is in contradiction with what is generally reported concerning hepatic steatosis induced by dietary disturbances in mammals but could be explained by the very specific conditions imposed by force-feeding. Despite this, force-feeding female ducks seems entirely feasible, provided that the final quality of the product is as good as that of the male ducks, which will remain to be demonstrated in future studies.

Dietary Amino Acid Source Elicits Sex-Specific Metabolic Response to Diet-Induced NAFLD in Mice.

SCOPE: Non-alcoholic fatty liver disease (NAFLD) is a sexually dimorphic disease influenced by dietary factors. Here, the metabolic and hepatic effects of dietary amino acid (AA) source is assessed in Western diet (WD)-induced NAFLD in male and female mice. METHODS AND RESULTS: The AA source is either casein or a free AA mixture mimicking the composition of casein. As expected, males fed a casein-based WD display glucose intolerance, fasting hyperglycemia, and insulin-resistance and develop NAFLD associated with changes in hepatic gene expression and microbiota dysbiosis. In contrast, males fed the AA-based WD show no steatosis, a similar gene expression profile as males fed a control diet, and a distinct microbiota composition compared to males fed a casein-based WD. Females are protected against WD-induced liver damage, hepatic gene expression, and gut microbiota changes regardless of the AA source. CONCLUSIONS: Free dietary AA intake prevents the unhealthy metabolic outcomes of a WD preferentially in male mice.

Evolution of oxidative stress markers in livers of ducks during force-feeding.

Mule ducks have been force-fed to develop a hepatic steatosis, also called "foie gras", which is similar to the non-alcoholic fatty liver disease (NAFLD) described in humans and mammals. However, in hepatic steatosis resulting from force-feeding of ducks, very little is known about the fine biochemical events that occur due to the enormous and very rapid increase in total lipids that mainly accumulate in hepatocytes. To begin to reduce this lack of knowledge associated with the development of this specific hepatic steatosis, liver samples were taken at different times to follow the overall biochemical transformation of the liver as well as different markers of oxidative stress, hypoxia and apoptosis. The results indicate that the lipid content increases rapidly in the liver throughout the force-feeding period while the protein content decreases. The amount of hydroxyproline remains constant indicating that no liver fibrosis develops during the force-feeding period. On the contrary, all the tested biomarkers of cellular oxidative stress increase rapidly but without any visible disorder in the coordination of paired activities. At the same time, hypoxia-inducible factors also increase indicating that a hypoxia situation is gradually occurring in hepatocytes. This leads, in addition to the lipotoxicity induced by the accumulation of lipids, to an increased number of liver cells to enter into apoptosis. A relative variability in the level of these cellular responses was also observed indicating that, probably, certain animals support the development of this steatosis differently. This leads us to imagine that the physiological status of these birds may differ widely for reasons that remain to be clarified.

Slightly different metabolomic profiles are associated with high or low weight duck foie gras.

Understanding the evolution of fatty liver metabolism of ducks is a recurrent issue for researchers and industry. Indeed, the increase in weight during the overfeeding period leads to an important change in the liver metabolism. However, liver weight is highly variable at the end of overfeeding within a batch of animals reared, force-fed and slaughtered in the same way. For this study, we performed a proton nuclear magnetic resonance (1H-NMR) analysis on two classes of fatty liver samples, called low-weight liver (weights between 550 and 599 g) and high-weight liver (weights above 700 g). The aim of this study was to identify the differences in metabolism between two classes of liver weight (low and high). Firstly, the results suggested that increased liver weight is associated with higher glucose uptake leading to greater lipid synthesis. Secondly, this increase is probably also due to a decline in the level of export of triglycerides from the liver by maintaining them at high hepatic concentration levels, but also of hepatic cholesterol. Finally, the increase in liver weight could lead to a significant decrease in the efficiency of aerobic energy metabolism associated with a significant increase in the level of oxidative stress. However, all these hypotheses will have to be confirmed in the future, by studies on plasma levels and specific assays to validate these results.

Using 2-DE for the differentiation of local chicken breeds.

The aim of this study was to apply a proteomic approach for the characterisation of local chicken breeds. The experiment involved a total of 29 males of three local Italian chicken breeds: Pépoi, Padovana and Ermellinata di Rovigo. Sarcoplasmic protein fractions of breast muscle were analysed by 2-DE. Image analysis followed by statistical analysis enabled to differentiate groups of individuals based on the similarities of protein expression. Individuals were distinguished into clusters and groups, corresponding to the breed of origin. Distances among individuals, calculated using data on spot volumes, were used to draw a neighbour-joining tree, showing clear individual and breed grouping. The most relevant spots regarding breed differentiation were detected; 11 were identified by MS revealing preliminary evidences on the mechanisms of the breed differentiation process. The results evidenced the ability of proteomic analyses for the characterisation of chicken breeds.

Roles of Estrogens in the Healthy and Diseased Oviparous Vertebrate Liver.

The liver is a vital organ that sustains multiple functions beneficial for the whole organism. It is sexually dimorphic, presenting sex-biased gene expression with implications for the phenotypic differences between males and females. Estrogens are involved in this sex dimorphism and their actions in the liver of several reptiles, fishes, amphibians, and birds are discussed. The liver participates in reproduction by producing vitellogenins (yolk proteins) and eggshell proteins under the control of estrogens that act via two types of receptors active either mainly in the cell nucleus (ESR) or the cell membrane (GPER1). Estrogens also control hepatic lipid and lipoprotein metabolisms, with a triglyceride carrier role for VLDL from the liver to the ovaries during oogenesis. Moreover, the activation of the vitellogenin genes is used as a robust biomarker for exposure to xenoestrogens. In the context of liver diseases, high plasma estrogen levels are observed in fatty liver hemorrhagic syndrome (FLHS) in chicken implicating estrogens in the disease progression. Fishes are also used to investigate liver diseases, including models generated by mutation and transgenesis. In conclusion, studies on the roles of estrogens in the non-mammalian oviparous vertebrate liver have contributed enormously to unveil hormone-dependent physiological and physiopathological processes.

Cellular markers of mule duck livers after force-feeding.

The "Foie gras" or fatty liver is the result of hepatic steatosis from nutritional origin and induced by the force-feeding of palmipeds. Despite identical rearing and force-feeding conditions of ducks from the same breed, different liver weights, within a range of 500 to more than 700 g, are generally observed at the time of evisceration. To better understand the determinism of this large variability in fatty liver weights, the activity of various metabolic pathways has been explored in 4 groups of steatotic livers differing by their weights. Different analyses were performed using biochemical assays on metabolites as well as ELISA tests or enzyme activity assays. The result showed that an increase in the final liver weight is always associated with a hypoxic response and even a severe hypoxia observed in livers with the highest weights (more than 650 g). This is also combined with a rise in the cellular oxidative stress level. In addition, for the heaviest livers (more than 700 g), signs of cell death by apoptosis were also observed, while others programmed cell death pathways, such as ferroptosis or necroptosis, seemed to be nonactive.

Proteomic Analysis of Two Weight Classes of Mule Duck "foie gras" at the End of an Overfeeding Period.

The weight of the liver is one of the important selection criteria in the quality of "foie gras". This factor is highly variable despite the fact that individuals are reared, overfed and slaughtered in the same way. In this study, we performed an analysis of the proteome profile of two weight classes of light (between 550 and 599 g) and heavy (more than 700 g) livers. For the analysis of the proteic extracts, a liquid chromatographic analysis coupled with mass spectrometry was carried out. In low-weight livers, aerobic energy metabolism, protein metabolism and lipid metabolism oriented toward export and beta-oxidation were overexpressed. On the contrary, high weight livers were characterized by anaerobic energy metabolism and a more active protein catabolism associated with cell apoptosis and reorganization of the cell structure.

Evolution of liver fattening and foie gras technological yield during the overfeeding period in mule duck.

In foie gras production the technological yield after the cooking process is one of the main issues of processors as it is closely linked to the cooking melting rate. This rate is subjected to strict laws and regulations since it directly affects the organoleptic and technological qualities of this gourmet product. The objective of the study was to better understand the liver fattening and the technological yield decrease during the overfeeding kinetics. A flock of 210 mule ducks was reared and then overfed during 12 D with 2 overfeeding programs; in the test group the amounts of corn in the first meals were higher than in the control group (+430 g during the whole period). Ducks were slaughtered at the end of the rearing period (D0, n = 15) and every other day (D2 to D12, n = 15 by group). Duck performances, anatomical dissections and physical and biochemical liver characteristics were registered. The performances were equivalent in the groups (P > 0.1). The evolution of the liver weight was then analyzed in detail in relation with the evolution of its biochemical composition. A two-step evolution occurred in the liver metabolism, first a main glycogen storage and then a strong lipid storage. A model to predict the liver weight was established with only BWs and feed intakes (R² = 0.83). The technological yield was determined on foie gras weighing more than 300 g (D6 to D12). The melting process was high during the last 2 D. The technological yield reached 72% at D12, for 758 g foie gras, and a strong negative correlation was observed with liver weight (-0.83; P < 0.001). A model to predict the technological yield was established with the liver weight and the liver color parameters (R² = 0.71). This study highlights the compromise between foie gras weight and its quality.

Effects of physical activities on biochemical and biomechanical properties of tendons in two commercial types of chickens.

The aim of our experiment was to study the effects of physical activities on the biochemical and biomechanical properties of tendons in 12 standard (S) broilers and 12 Label Rouge (LR) chickens. In the two types of birds no differences were found between control and active birds for body weights. Gastrocnemius (Gas) tendon and Pectoralis minor (Pm) tendons were harvested and processed for passive stretch tests prior to cooking or not. Some biochemical parameters also were determined. Results showed that total collagen content in Gas tendon was significantly higher in active than in control birds. However, no significant changes were found in collagen solubility in LR tendons while these values were increased in S ones. Active birds showed greater sGAGs content than control ones. Ultimate load was found to be significantly higher in active birds than in control. Deformability (defined by Poisson's ratio) of raw and heated at 80 degrees C Gas tendons increases in active groups because Poisson's ratio decreases. Physical activities also increase the rigidity (defined by elastic modulus) of raw and heated at 80 degrees C Gas tendons because elastic modulos values increase. Physical activity was not able to modify stiffness or maximum stress values in raw or heated at 80 degrees C Gas tendons from broilers whereas these two parameters were found to be slightly higher in active group from LR chickens only in raw tendons. All the biomechanical results recorded in Pm tendons from both types of chickens were not significantly different between control and active birds. A significant correlation was found between the total collagen content and stiffness in Gas tendon from LR active birds.

Two-Dimensional Zymography of Proteases from Steatotic Duck Liver.

Protease activity present in liver cells with steatosis can be electrophoretically characterized. Zymographic techniques allow semi-quantitative results, successfully detecting cathepsin and metalloprotease activity using polyacrylamide gels copolymerized with gelatin and quantified by densitometry. By using specific inhibitors, the identity of the proteases can be confirmed. 2D zymography allows the determination of both M r. and pI of the metalloprotease and cathepsin activity present in the homogenates. The analysis of liver proteases activities in force fed ducks may elucidate the mechanisms behind steatosis development.

Relationship between Proteolytic Activities and Cooking Loss Variability in Liver Issued from Force-Fed Mule Ducks.

We investigated liver protease activities to better understand the mechanisms responsible for losses during the cooking of Mule ducks' fatty livers. Fatty livers of similar masses (520 g) were randomly selected among a large flock of overfed male Mule ducks and divided into two groups on the basis of their cooking loss rate, L+ (high cooking losses, 36%) and L- (low cooking losses, 21%). In addition to dry matter and total lipid contents, main hepatic protease (matrix metalloproteinase 2, cathepsins, and calpains) activities were measured by zymography. The results show that L+ samples present higher total relative proteolytic activity (6.15 ± 1.07 vs 4.46 ± 0.52 for L+ and L- samples, respectively) and total lipid content (61.0 ± 3.03% vs 53.7 ± 3.22% for L+ and L- samples, respectively) than L- samples. The results imply that proteases could be involved in the fragilization of the hepatocyte structure, resulting thus in higher cooking losses in L+ samples.

The effects of overfeeding on myofibre characteristics and metabolical traits of the breast muscle in Muscovy ducks (Caïrina moschata).

The aim of this experiment was to study the consequences of precise feeding on the myofibre characteristics and metabolic traits of the breast muscle (Pectoralis major, Pm) of Muscovy ducks. Twenty-four samples of breast muscle, without skin or subcutaneous fat, from two groups of ducks, control and overfed respectively, were collected at 14 weeks of age. We assayed different chemical (water content, crude proteins, total lipid ashes, total and thermosoluble collagen), biochemical (activities of the CS, LDH and beta-HAD enzymes), histological (muscle fibre typing and intramuscular adipocyte measurements) and technological (drip and cooking losses, texture) determinations. At the force-feeding period, the overfed ducks weighed 6366 g and the control ducks 4606 g of body weight. In the PM muscle, some modifications of the biochemichal parameters and enzyme activities were observed but neither the shear force nor the histological characteristics of the breast muscle were affected by the fattening treatment. The overfed birds had an increased total lipids content (correlated to an increase in the intramuscular area occupied by the adipocyte) and a different fatty acid profile as the result of a higher energy feed intake. The lipids of the Pm muscle of the overfed ducks contained more C16:0, C16:1n-7 and C18:1n-9, but less C18:0, C18:2n-6 and C20:4n-6 than the control birds. These results show that in response to high energy feeding, the muscle is able to respond quickly on a metabolic basis (by increasing the activities of the oxydative enzymes) without changing its typology or morphology. Additionally, fattening was correlated to a degradation in the technological qualities of the breast muscle, especially an increase in the cooking losses.

Genetic parameters of meat technological quality traits in a grand-parental commercial line of turkey.

Genetic parameters for meat quality traits and their relationships with body weight and breast development were estimated for a total of 420 male turkeys using REML. The birds were slaughtered in a commercial plant and the traits measured included pH at 20 min (pH20) and 24 h post-mortem (pHu) and colour of the breast and thigh meat. The heritabilities of the rate and the extent of the pH fall in the breast muscle were estimated at h2=0.21 +/- 0.04 and h2=0.16 +/- 0.04, respectively. Heritabilities ranging from 0.10 to 0.32 were obtained for the colour indicators in the breast muscle. A marked negative genetic correlation (rg=-0.80 +/- 0.10) was found between pH20 and lightness (L*) of breast meat, both traits corresponding to PSE indicators. The pH20 in the thigh muscle had a moderate heritability (h2=0.20 +/- 0.07) and was partially genetically related to pH20 in the breast muscle (rg=0.45 +/- 0.17). Body weight and breast yield were positively correlated with both initial and ultimate pH and negatively with the lightness of breast meat.