E. Baéza

Intramuscular fat content in meat-producing animals: development, genetic and nutritional control, and identification of putative markers.

Intramuscular fat (IMF) content plays a key role in various quality traits of meat. IMF content varies between species, between breeds and between muscle types in the same breed. Other factors are involved in the variation of IMF content in animals, including gender, age and feeding. Variability in IMF content is mainly linked to the number and size of intramuscular adipocytes. The accretion rate of IMF depends on the muscle growth rate. For instance, animals having a high muscularity with a high glycolytic activity display a reduced development of IMF. This suggests that muscle cells and adipocytes interplay during growth. In addition, early events that influence adipogenesis inside the muscle (i.e proliferation and differentiation of adipose cells, the connective structure embedding adipocytes) might be involved in interindividual differences in IMF content. Increasing muscularity will also dilute the final fat content of muscle. At the metabolic level, IMF content results from the balance between uptake, synthesis and degradation of triacylglycerols, which involve many metabolic pathways in both adipocytes and myofibres. Various experiments revealed an association between IMF level and the muscle content in adipocyte-type fatty acid-binding protein, the activities of oxidative enzymes, or the delta-6-desaturase level; however, other studies failed to confirm such relationships. This might be due to the importance of fatty acid fluxes that is likely to be responsible for variability in IMF content during the postnatal period rather than the control of one single pathway. This is evident in the muscle of most fish species in which triacylglycerol synthesis is almost zero. Genetic approaches for increasing IMF have been focused on live animal ultrasound to derive estimated breeding values. More recently, efforts have concentrated on discovering DNA markers that change the distribution of fat in the body (i.e. towards IMF at the expense of the carcass fatness). Thanks to the exhaustive nature of genomics (transcriptomics and proteomics), our knowledge on fat accumulation in muscles is now being underpinned. Metabolic specificities of intramuscular adipocytes have also been demonstrated, as compared to other depots. Nutritional manipulation of IMF independently from body fat depots has proved to be more difficult to achieve than genetic strategies to have lipid deposition dependent of adipose tissue location. In addition, the biological mechanisms that explain the variability of IMF content differ between genetic and nutritional factors. The nutritional regulation of IMF also differs between ruminants, monogastrics and fish due to their digestive and nutritional particularities.

Variations in chicken breast meat quality: implications of struggle and muscle glycogen content at death.

1. Pectoralis major (P. major) muscle pH and meat quality traits were studied in relation to bird response to ante-mortem stress in three chicken lines: a fast-growing standard line (FGL), a slow-growing French ‘Label Rouge’ line (SGL) and a heavy line (HL). Ninety-nine birds of the three genetic types were slaughtered at their usual marketing age (6, 12 and 6 weeks for FGL, SGL and HL birds, respectively) on the same day. The birds of each line were divided into three different ante-mortem treatment groups: minimum stress (shackling for 10 s) (C), shackling for 2 min (SH) and acute heat plus shackling stress (exposure to 35°C for 3·5 h and shackling for 2 min before stunning) (H + SH). 2. Regardless of chicken line, wing flapping duration (WFD) between hanging and stunning was strongly negatively related to P. major muscle pH at 15 min post-mortem. It was also moderately negatively related to P. major muscle glycolytic potential (GP), which represents glycogen level at death. Increasing WFD induced an increased ultimate pH (pHu) only in HL. The consequences of increased WFD for breast meat traits were dependent on the chicken line: it induced lower L* and b* and higher a* and drip loss in SGL while it only increased breast a* in HL birds. By contrast, WFD variations did not alter breast meat quality traits of FGL birds. Regardless of the chicken line, increased GP was associated with lower pHu and higher L* and drip loss. In SGL, it also increased b* and decreased curing–cooking yield of breast meat. 3. Struggling activity on the shackle line and muscle glycogen content at death could partly explain line and pre-slaughter variations in breast meat pH and quality traits. The water holding capacity of the raw and cooked meat was impaired by long shackling in the case of SGL birds while it was barely affected by ante-mortem conditions in the two standard lines. In conditions which minimised bird struggling (C), SGL and FGL birds had meat with a better water holding ability than that of broilers from the heavy line. However, when broilers were subjected to SH or H + SH conditions, the breast meat water holding capacity of SGL birds was lowered to the same level as that of the heavy line birds.

Behavioural and physiological responses of three chicken breeds to pre-slaughter shackling and acute heat stress.

1. The aim of this study was to compare the behavioural and physiological responses to hanging and acute heat stress in three different chicken breeds. Chicks were obtained from a slow-growing French ‘Label Rouge’ line (SGL), a fast-growing standard line (FGL) and a heavy line (HL). The SGL, FGL and HL birds were slaughtered at their respective market ages of 12, 6 and 6 weeks, in an attempt to achieve similar body weights. Before stunning, birds were either shackled by their legs on the moving line for 2 min (shackling stress: SH) or placed in a room at 35°C and 60% of humidity for 3·5 h and then shackled for 2 min (acute heat stress plus shackling: H + SH) or subjected to minimal stress by shackling for 10 s before stunning (control group: C). 2. Bird physiological responses to the three pre-slaughter treatments were estimated by measuring blood corticosterone, glycaemia, creatine kinase activity, acid–base status and electrolyte concentration as well as lactate content and glycolytic potential in the breast (Pectoralis major) and thigh (Ilio tibialis) muscles. Behavioural responses to shackling stress were evaluated by measuring wing flapping duration, straightening up attempts and vocalisations. 3. Blood corticosterone was higher in SH and H + SH groups than in the C group, regardless of genotype. The struggling activity on the shackle line differed among chicken breeds. It was more intense and occurred more rapidly after hanging in the SGL birds than in both other breeds. Furthermore, SGL struggling activity was not affected by hanging duration while it increased with hanging duration in FGL and HL birds. 4. Wing flapping duration was negatively correlated with blood pH, bicarbonate concentration and positively correlated with breast muscle lactate content, indicating that struggling stimulated ante-mortem glycolysis activity in breast muscle. Acute heat stress affected blood Ca2+ and Na+ concentration and increased glycaemia and glycolytic potential of thigh muscle. 5. Both acute heat stress and shackling before slaughter were experienced as stressful events by all types of birds.

Effect of selection for improved body weight and composition on muscle and meat characteristics in Muscovy duck

1. For 25 years, the Muscovy duck has been selected for greater body weight at slaughter age, higher meat yield and lower fat content. The aim of this study was to analyse the effects of such selection on the biochemical and technological characteristics of duck meat. 2. A line which has been maintained without any selection since 1975 (control line) and a heavy line now commercialised (selected line) were reared under the same conditions. We compared growth performances, dressing and dissection yields and biochemical and technological characteristics of duck meat at 8, 10 and 12 weeks of age. 3. Our results showed that an indirect effect of selection was to increase the duck growth rate. At a given age, muscle weights and yields as well as carcase fat content were higher for selected ducks. In the selected line, muscle fibres were larger and the collagen content in muscles lower. 4. The Sartorius muscle in ducks from the selected line exhibited a higher percentage in numbers of white type IIb fibres to the detriment of red type IIa fibres. Its glycolytic enzyme activity was higher and its oxidative enzyme activity lower. 5. In breast muscle, selection decreased the rate of post-mortem decline in pH value, increased protein and mineral contents and decreased moisture. It did not affect fibre type, colour, haem pigment and lipid contents, collagen solubility or drip loss. 6. In conclusion, the effects of Muscovy duck selection on meat quality were relatively moderate.

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.

Effects of the rate of muscle post mortem pH fall on the technological quality of turkey meat

1. This experiment evaluated the influence of the rate of post mortem pH fall on the processing ability of turkey meat. 2. Four hundred and twenty male turkeys from a selected pure line (grand-parental female line, BUT Ltd) were slaughtered at 16 weeks of age in a commercial plant and pH was measured in the Pectoralis superficialis (PS) and Ilio tibialis (IT) muscles, at 20 min post mortem . Three groups of PS muscle differing in pH 20 and two groups of IT muscle differing in pH 20 were constituted and processed as cured-cooked white meat and turkey ham, respectively. 3. The technological yield was lower in the groups showing the lowest pH 20 (97.4% at pH 20 5.90 vs 98.6 and 98.3% at pH 20 6.24 and 6.55, respectively, for white meat and 97.2% (pH 20 6.28) vs 98.3% (pH 20 6.56) for turkey hams). The groups showing the lowest pH 20 also showed higher drip loss in commercially packed products. 4. Acceptability tests of processed products were carried out in the commercial plant. Texture and taste of white meat were better in the highest pH group but the overall impression was similar in the lowest and the highest pH groups (mean scores of 4.2 and 4.1, respectively), due to better colour in the former (mean scores of 4.4 for the lowest pH 20 group vs 4.0 and 3.9 for the medium and highest pH 20 groups, respectively). 5. For turkey hams, the meat processed from the highest pH group got the best score for all items.

Effects of age and sex on the structural, chemical and technological characteristics of mule duck meat

1. The aim of the study was to analyse the effect of age and sex on the chemical, structural and technological characteristics of mule duck meat. 2. Ten males and 10 females were weighed and slaughtered at 8, 10, 11, 12 and 13 weeks of age. Weight, pH value, colour, tenderness and juice loss of breast muscle were determined. 3. The activities of 3 enzymes (citrate synthase, β -hydroxyacyl CoA dehydrogenase, lactate dehydrogenase) which indicate muscular metabolic activity were assayed. 4. Chemical composition (moisture, lipids, proteins, minerals, lipid and phospholipid classes, fatty acid composition) of breast muscle was analysed. 5. Fibre type, fibre type percentage and cross-sectional areas were determined using histochemistry and an image analysis system. 6. For growth performance and muscular structure, the ideal slaughter age of mule ducks is 10 weeks of age. Chemical and technological analysis indicated that muscular maturity in Pectoralis major was reached at 11 weeks of age, but, at this age, breast lipid content is high. Moreover, after 10 weeks of age, food costs rapidly increased. 7. Lastly, sexual dimorphism for body weight is minor. In this study, at any given age, no significant differences between males and females were shown. Thus, it is possible to rear both sexes together and to slaughter them at the same age.

Harmonization of methodologies for the assessment of poultry meat quality features

Many different methods measuring meat quality traits are available which are based on different principles, and instruments and/or probes. In view of the complexity of meat processes after slaughter and quality trait determination, it is not surprising that the results obtained in different studies and laboratories are not always in agreement. For comparison of results it is therefore necessary to keep strictly to measurable specifications, which is why standardisation is indispensable. The Working Group 5 Poultry Meat Quality group of the WPSA European Federation has been asked to produce a document which would serve as a common base methodology that would permit comparison between research projects carried out by different groups, based on international research programmes. This paper represents the first step of this work including chemical (moisture, total lipids, proteins, ash, fatty acid composition, cholesterol, susceptibility to oxidation, amino acids, collagen and pigments) and physical traits (pH, R-value, colour, water holding capacity, texture and sarcomere length). For the evaluation of chemical composition, there are standard methods available which are largely adopted in the majority of published papers. However, there is still a need to standardise methods for determining the physical traits to facilitate comparisons between studies and to provide reference values.

Gene expression and protein content in relation to intramuscular fat content in Muscovy and Pekin ducks

Independent of their nutritional condition, Pekin ducks always exhibit higher i.m. fat content than Muscovy ducks. To understand this difference between species, the expression level of genes involved in lipid metabolism was analyzed in the pectoralis major muscle of Pekin and Muscovy ducks ad libitum-fed or overfed. The lipoprotein lipase (LPL) gene expression was not different between species and not influenced by overfeeding. The protein content for LPL was higher in Pekin ducks than in Muscovy ducks when birds were ad libitum-fed, whereas in overfed ducks, we found no difference between species. Adipocyte fatty acid-binding protein (A-FABP) gene expression and protein content were higher in Pekin ducks than in Muscovy ducks for each nutritional condition (suggesting a higher intracellular transport within i.m. adipocytes of fatty acids mainly provided by liver for this species). Overfeeding did not affect the expression of genes involved in oxidation [carnitine palmitoyl transferase 1A (CPT1A), cytochrome-c oxidase 4 (COX4), succinyl-coenzyme A:3-ketoacid coenzyme A transferase (SCOT)] but increased the expression of fatty acid synthase (FAS) involved in lipogenesis. For all nutritional conditions, Pekin duck exhibited higher expression levels of CPT1A, COX4, SCOT, and FAS than Muscovy ducks. Results for mRNA SCOT suggested that the muscles of Pekin ducks use ketone bodies as an energy source. In conclusion, i.m. lipogenesis could contribute to the i.m. fat, particularly in Pekin ducks.

The Loss of Adipokine Genes in the Chicken Genome and Implications for Insulin Metabolism

Gene loss is one of the main drivers in the evolution of genomes and species. The demonstration that a gene has been lost by pseudogenization is truly complete when one finds the pseudogene in the orthologous genomic region with respect to active genes in other species. In some cases, the identification of such orthologous loci is not possible because of chromosomal rearrangements or if the gene of interest has not yet been sequenced. This question is particularly important in the case of birds because the genomes of avian species possess only about 15,000 predicted genes, in comparison with 20,000 in mammals. Yet, gene loss raises the question of which functions are affected by the changes in gene counts. We describe a systematic approach that makes it possible to demonstrate gene loss in the chicken genome even if a pseudogene has not been found. By using phylogenetic and synteny analysis in vertebrates, genome-wide comparisons between the chicken genome and expressed sequence tags, RNAseq data analysis, statistical analysis of the chicken genome, and radiation hybrid mapping, we show that resistin, TNFα, and PAI-1 (SERPINE1), three genes encoding adipokines inhibiting insulin sensitivity, have been lost in chicken and zebra finch genomes. Moreover, omentin, a gene encoding an adipokine that enhances insulin sensitivity, has also been lost in the chicken genome. Overall, only one adipokine inhibiting insulin sensitivity and five adipokines enhancing insulin sensitivity are still present in the chicken genome. These genetic differences between mammals and chicken, given the functions of the genes in mammals, would have dramatic consequences on chicken endocrinology, leading to novel equilibriums especially in the regulation of energy metabolism, insulin sensitivity, as well as appetite and reproduction.