Post mortem development of meat quality as related to changes in cytoskeletal proteins of chicken muscles.

1. A procedure was developed to separate high and medium molecular weight myofibrillar proteins from chicken muscular tissue with a high resolution by flat bed sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and subsequent detection by either a general protein stain or Western blotting. These procedures were used to analyse the degradation process of cytoskeletal proteins in chicken breast and leg muscles during meat ageing. 2. This study demonstrates the degradation of all the examined cytoskeletal proteins: titin, nebulin and desmin as well as vinculin, a protein component of the costamere structure. All the examined proteins were found to be degraded during ageing of chicken breast and leg muscles. 3. Degradation of titin, nebulin and desmin started at 3 h post mortem in breast muscle. Intact titin and nebulin disappeared within 1 d. Intact desmin and vinculin were not detectable after 3 d post mortem. In leg muscle, the degradation process of all the examined proteins evolved much more slowly than in breast chicken muscles. 4. The changes observed in shear force, myofibrillar fragmentation and cooking loss were related to changes in cytoskeletal proteins and used to identify marker proteins or degradation products for the purpose of monitoring the development of meat ageing. The ageing process was faster in breast muscle than in leg muscle. 5. Significant correlations were found between degradation processes of titin, nebulin, and desmin and shear force, as well as myofibril fragmentation index of breast and leg muscles.

Effects of feed deprivation and electrical, gas, and captive needle stunning on early postmortem muscle metabolism and subsequent meat quality.

The general method for stunning poultry before slaughter is by immersion of a chicken's head into an electrified waterbath. This method results in carcass and meat quality deficiencies. The major problems are hemorrhages and a delay in onset of rigor mortis, which increases the risk of cold shortening with early deboning. In two experiments, this study examines the early postmortem metabolism in the breast muscle and its effect on ultimate meat quality. The first experiment describes the effects of 5 h feed deprivation on the availability of glycogen from the liver and the breast muscle, of waterbath and head-only electrical stunning on pH and metabolite levels up to 6 h in unprocessed muscle, and the consequences on meat quality. The second experiment compares the same measurements after waterbath and head-only electrical stunning, CO2/O2/N2 and Ar/CO2 gases, and captive needle stunning. Metabolic degradation halted after 6 h without processing or after 4 h under conventional conditions after waterbath and CO2/O2/N2 stunning. With other stunning methods, this occurrence is at a faster rate, largely depending on muscle activity. Muscle glycogen does not need to be exhausted for energy generation to cease. If glycogen is a limiting factor, as found with head-only stunning, pH drops too rapidly and affects water-holding capacity and color. Hemorrhage scores were higher with electrical stunning than with other stunning methods. Gas stunning affected color and, to a lesser extent, water-holding capacity. Captive needle stunning scored between gas and electrical stunning on most measurements.

Induction and characterization of multianalyte antibodies against penicillins in egg yolk.

Antibodies against penicillins were induced in eggs of laying hens after immunization with 6-aminopenicillanic acid (6-APA) coupled to key-hole limpet hemocyanin (KLH). Development of the antibody titer was monitored by an indirect enzyme-linked immunosorbent assay (ELISA), with 6-APA coupled to ovalbumin as antigen for coating microtiter plates. Different characteristics (time course, affinity) of the immune reaction were observed by testing eggs of individual hens. Titer values varied between 150 and 2000. Antibodies were isolated by polyethylene glycol precipitation and affinity chromatography, using a hapten sorbent with 6-APA as ligand. Glycine buffer, pH 3.0, was used to elute the immunoglobulins. Antibody specificity was determined in a competitive ELISA with 7 penicillins and the cephalosporin cephalexin as competitors. Cross reactivities for the penicillins were between 100 and 290% (6-APA = 100%). Cephalexin cross reacted only marginally (3%).

Endogenous proteolytic enzymes in chicken muscles. Differences among strains with different growth rates and protein efficiencies.

The theory that net muscle growth is, at least partly, regulated by catabolic factors has been tested in order to set up an animal model to study meat aging and post-mortem tenderization. Male and female chickens of a layer strain (White Leghorn), a commercial broiler strain (Ross), and two experimental broiler lines (designated GL and FC) were used to estimate differences in proteolytic enzyme activities in the breast muscles. The GL and the FC lines were selected for high body weight gain and high feed efficiency, respectively. At 6 wk of age the birds were slaughtered and the activities of endogenous proteinases and their specific inhibitors in breast muscles measured. The Leghorns showed significant differences in all traits compared with the three broiler genotypes. Within the broiler types, FC birds tended in the direction of the Leghorns and GL birds in the opposite direction. Ross birds were intermediate between FC and GL birds. All types and sexes differed significantly in slaughtering weight. Feed conversion ratio and protein conversion ratio were highest for Leghorns. The FC birds showed the lowest feed conversion. Ross and GL birds showed intermediate values. The Leghorns showed higher calpain activities and lower calpastatin activity than the three broiler genotypes. The FC broilers showed intermediate calpain and calpastatin activities but higher cathepsin H and total cystatin values. The GL broilers showed lower cathepsin B, D, and H activities. In all cases the Ross broilers showed intermediate values. From these figures it is concluded that the strains of birds used in this study can be used as a natural source of variability to study the mechanisms involved in post-mortem proteolytic degradation and thus in the study of muscle tenderization and meat aging. It is also concluded that it could be very interesting to study the behavior of the different proteolytic systems more carefully in relation to muscular growth characteristics and compare them to anabolic factors involved in muscle growth.

Ascites in broilers. 2. Disturbances in the hormonal regulation of metabolic rate and fat metabolism.

Effects of different broiler stocks, ambient temperatures (Ta), dietary energy content (AME), and dietary levels of unsaturated fat on plasma thyroid and growth hormone concentrations and energy metabolism were studied. An experiment with a 2 x 2 x 2 x 2 factorial split-plot arrangement of treatments with 96 groups of 12 male broilers each was performed. Blood samples were taken at 3, 4, and 5 wk of age. Energy metabolism parameters were determined over an entire period from 1 to 5 wk of age. Chickens from a line selected for fast growth rate and low feed conversion ratio but also more sensitive to heart failure syndrome (HFS) and ascites (Line SS) than commercial birds (Line BC) exhibited the greatest responses to experimental factors. Differences in levels of plasma thyroxine (T4), triiodothyronine (T3), reverse triiodothyronine (rT3), and growth hormone (GH) between stocks at different ages were highly dependent on Ta and dietary fat content. Differences in heat production per metabolic weight, percentage of retained fat energy in retained energy, and efficiency of AME intake for retained energy between stocks corresponded to differences in hormone levels. High-fat diets (polyunsaturated fatty acids) inhibited the extra thyroidal conversion of T4 to T3 in both stocks. Differences between stocks in T3 and rT3 levels in plasma indicated that BC birds (in contrast to SS birds) were better able to compensate for an inhibited T4 conversion to T3 by producing more T4. Overall results suggest that the occurrence of HFS and ascites in SS birds could be initiated independently by different factors. These factors might be a limited thyroid hormone production and a lower capacity for oxygen consumption. An inverse relationship between T3 and GH levels found in particular combinations of experimental factors, together with changes in fat deposition, support published concepts about the positive effects of T3 on lipogenesis and GH on lipolysis.