Mitochondrial function in turkey skeletal muscle--impact on meat quality.

1. M. iliotibialis (MIT) and M. pectoralis (MP) of the BUT Big 6 and Kelly BBB turkey were characterised with respect to physical properties, mitochondrial function, metabolic state, morphology and meat quality. 2. Mitochondrial enzyme activity and respiration rates in MP declined with increasing age while glycolytic enzyme activity remained nearly constant. 3. There were no major differences between BUT Big 6 and Kelly BBB with respect to histological, histochemical or biochemical variables. In spite of the greater adult weight of BUT Big 6, body proportion was equal in both strains. 4. In agreement with the histochemical findings MIT showed higher oxidative capacities, while glycolytic enzyme activity was higher in MP. 5. Pyruvate was the best substrate for oxidative phosphorylation in MIT, but not in MP. Pyruvate dehydrogenase (PDH) activity was up to 15 times less in MP and blood lactate was correlated with intramuscular pH. 6. Turkey breast muscle was restricted in its ability to oxidise pyruvate, especially in those animals that tended to develop intramuscular acidosis post mortem. 7. It is concluded that the in vivo metabolic environment results in acidosis and impaired meat quality, at least in turkey M. pectoralis.

Effect of long-term heat exposure on peripheral concentrations of heat shock protein 70 (Hsp70) and hormones in laying hens with different genotypes.

1. The effect of long-term, moderate heat stress (30 to 32 degrees C) on heat shock protein 70 (Hsp70) concentration in mononuclear blood cells and plasma concentrations of 3,5,3'-triiodothyronine (T3) and corticosterone in laying hens was investigated. 2. Three groups of 48 hens each (Ethopian line [Angete Melata, Na], New Hampshire [NH], F1 cross [Na x NH]) were divided into an experimental group (24 each) and a control group (24 each, ambient temperature 18 to 20 degrees C), respectively. All hens were kept in individual cages up to an age of 68 weeks and performance data were recorded. 3. Blood samples were taken from the wing vein of 12 hens from each group at weeks 22, 38, 51 and 65 (12 hens x 3 lines x 2 treatments). Mononuclear blood cells were isolated and Hsp70 concentrations were determined by Western Blot analysis with a monoclonal anti-Hsp70 antibody. T3 and corticosterone were measured with commercially available ELISA and RIA kits, respectively. 4. The moderate heat stress caused significantly increased Hsp70 levels compared with the control groups in weeks 51 and 65. However, the responses of the lines were not uniform at different ages. 5. In contrast, T3 levels were significantly decreased in stressed birds regardless of line and age. There was no effect of treatment and line on corticosterone levels during the experimental period. 6. Our results indicate that Hsp70 and T3 levels are affected by mild heat stress applied over a long period but are both involved in independent mechanisms of acquisition of thermotolerance. Further investigations are necessary to clarify whether the observed differences in Hsp70 response between the genotypes are indicators for differences in thermotolerance.

Isolation of expressed sequence tags of skeletal muscle of neonatal healthy and splay leg piglets and mapping by somatic cell hybrid analysis.

We have isolated 14 differentially displayed and 10 further expressed sequence tags (ESTs) from Musculus biceps femoris of newborn healthy and splay leg piglets. By comparison with EMBL/GenBank data we could identify nine porcine homologues to human genes (TATA box binding protein associated factor B TAF1B; B-cell CLL/lymphoma 7B BCL7B; pyruvate dehydrogenase kinase, isoenzyme 4 PDK4; ribosomal protein S10 RPS10; SPARC-like 1 SPARCL1; epithelial protein lost in neoplasm beta EPLIN; N-myc downstream-regulated gene 2 NDRG2; pleiomorphic adenoma gene like 2 PLAGL and, BCL-2 associated transcription factor short form BTFS). Eight fragments correspond to uncharacterized ESTs and 7 ESTs had no significant match with database sequences. These data provide the first expression profiles in skeletal muscle of neonatal piglets and are a basis for candidate gene investigations for congenital splay leg in piglets. Eleven ESTs were physically mapped to porcine chromosomes 1, 3, 4, 5, 7, 8, 9 and 10 and contribute to the comparative map of humans and pigs.

Low microsatellite variation in laboratory gerbrils.

The Mongolian gerbil has become a model organism of increasing importance for the understanding of aging, epilepsy, the process of domestication or sociobiological questions. We report the development and characterization of the first nine polymorphic dinucleotide repeat loci in this species. Average observed heterozygosity and allele number of laboratory animals measured 0.136 (SE = +/-0.065) and 1.78 (SE = +/-0.278) compared to 0.761 (SE = +/-0.025) and 9.2 (SE = +/-0.57) found for a reference group of wild gerbils. The extreme low genetic variation observed in laboratory animals is caused by several severe population size bottlenecks due to the initial founder event and the later establishment of subpopulations. Reduced levels of allelic polymorphism in experimental animals hamper genetic mapping or parental studies. Therefore experiments relying on kinship analyses have to be carried out on wild animals. Estimates of genetic identity and parental exclusion were calculated as Pid = 2.8 x 10(-12) and Pex > 0.999 in wild gerbils. Laboratory gerbil strains show the expected high degree of genetic similarity. However, significant allele frequency differences (P < .001) between American and European gerbils at some microsatellite loci may still allow discrimination between breeding lines.

Immunohistochemical detection of components of the insulin-like growth factor system during skeletal muscle growth in the pig.

The insulin-like growth factor (IGF) system plays an important role in postnatal somatic and skeletal muscle growth in pigs. There is little information on the occurrence and distribution of components of the IGF system in postnatal porcine skeletal muscle. IGF-I, IGF receptor 1 (IGF1R) and the IGF-binding proteins IGFBP-1 and -3 in longissimus dorsi and triceps brachii were localized in muscle biopsies from 12 commercially crossbred pigs aged from 28 to 199 days as well as from the sire generation, by immunohistochemistry. Plasma IGF-I concentrations were also determined using radio-immunoassays. Unlike other species, IGF-I was localized in porcine skeletal muscle fibres. Staining intensity correlated with the highest plasma IGF-I levels and phases of intensive muscle growth from the 11th to 22nd week. The pattern of IGF1R immunostaining, which was strong, correlated with that of IGF-I, IGF1R was also localized in endomysial tissues. IGFBP-1 was not detected within muscle fibres, but was found in the endomysium and vessel walls, while IGFBP-3 was localized with IGF-1 and its receptor. Higher magnification revealed that IGF1R, IGFBP-3 and probably IGF-I appeared in the tubular system. Inhibitory as well as stimulating controls of IGFBP-1 and -3 on IGF functions are discussed, which may maintain a balance between autocrine growth promoting activities of IGF-I and IGF1R.

[Cell count and differential cell count in goat milk--variability and influencing factors]. / Zellgehalt und Differentialzellbild der Ziegenmilch--Variabilität und Einflussfaktoren.

In a total of 413 dairy goats of different breeds from five herds (milk performance 550-1000 kg) somatic cell counts and cell types were investigated in fore milk samples from udder halves, in composed udder and bulk milk samples. Cytoplasmatic particles in the milk were not counted. Three herds were investigated monthly during two years. At every sampling a clinical examination of the mammary gland (fore milk, teats, gland tissue, lymphatic nodes of the udders) and a bacteriological examination of the milk samples were carried out. In all herds the clinical health status of the udders was good. Independent of the sample type the mean SCC was about 1000 x 10(3)/ml with a wide range between several millions and < 100,000/ml. A seasonal influence on the monthly minima and maxima of the bulk milk SCC was evident, the lower SCC being recorded mainly during April to October. Increasing lactation numbers and months are connected with increasing SCC. Goats with high milk SCC at the begin of the lactation tended to maintain their high SCC levels as the lactation progressed. Limit values for SCC in goat milk should take into account the influence of the kidding season of the herds upon the dynamics of the bulk milk SCC. Polymorphonuclear leukocytes were the main cell type in goat milk (around 40%), their number tended to increase as total SCC increased, in particular in samples with cell counts above one million. Likewise, the differences between SCC in the milk of corresponding udder halves increased significantly as one of the halves showed more than one million somatic cells. Defining limit values for SCC in goat milk this should be taken into account.

Ex vivo investigation of blood monocyte and platelet behaviour in pigs maintained on an atherogenic diet.

Domestic pigs aged 4 months were fed for 16 weeks an atherogenic diet rich in cholesterol and saturated fatty acid. The increase of plasma cholesterol and triacylglycerol levels was found to be accompanied by a significant increase in the number of blood monocytes and platelets when compared to control animals. Furthermore, the atherogenic diet produced a small but significant reduction in the blood monocyte phagocytic capacity and adhesion to plastic surface. No significant differences between both groups were found when spontaneous platelet aggregation in whole blood was studied. However, platelets from pigs fed the atherogenic diet had a smaller mean cell volume compared to controls. The results indicate than an atherogenic diet may affect blood monocytes and platelets in pigs.

Abnormalities in the regulation of blood platelet free cytosolic calcium in malignant hyperthermia. II. Pig platelets.

Since 1966 the domestic pig has served as the animal model in Malignant Hyperthermia (MH) research [1]. The use of genetically well-defined pigs rendered it possible to test the method for diagnosing MH-susceptibility of patients presented in the preceding paper. Thus, the effect of halothane on intracellular calcium movements was studied in Quin-2- and chlorotetracycline-loaded pig platelets. In 'Ca(2+)-free' suspensions the resting level of free cytosolic Ca2+ was about 60 nM. In contrast to the results with human platelets there were no significant differences between pig genotypes either in the absence or in the presence of external calcium. After addition of halothane, a mobilization of intracellular membrane-bound calcium can be observed. However, the calcium mobilization is not accompanied by a marked increase in fluorescence intensity of Quin-2-loaded platelets. Thus, in the absence of external calcium, halothane produces only a slight increase in free cytosolic Ca2+. Nevertheless, the calcium rises measured in platelets from affected animals were statistically significantly higher than those from normal subjects. However, in the presence of 1 mM external calcium, a rapid increase in free cytosolic calcium can be detected after halothane addition. This suggests that halothane causes a marked, dose-dependent increase in Ca2+ permeability of the plasma membrane. Compared to the control group, significantly enhanced calcium permeability was found, not only in homozygous positive pigs, but also in heterozygous animals.

[Phospholipid content of subcellular structures in skeletal muscles after halothane loading in Landrace swine in relation to mating variants and genotype]. / Phospholipidgehalt subzellulärer Strukturen im Skelettmuskel nach Halothanbelastung bei Schweinen der Landrasse in Abhängigkeit von Anpaarungsvarianten und Genotyp.

Membraneous phospholipids of subcellular structures were determined from the musculature of German Landrace pigs of the GDR, following exposure to halothane. Mating variants A (H+ male X H+ female), B (H+ male X H- female), and C (H- male X H+ female) were used for positive responders (MHS), while variants B, C, and D (H- male X H- female) were used for negative responders (MHN). Four phospholipid fractions were recorded from the muscle samples for mitochondria and microsomes (according to SR section). Differences between the MHS and MHN groups for the above fractions and without consideration of mating variants and genotype were not observed, although unambiguous responses were exhibited by all animals, either positive or negative to halothan. Significant differences with regard to the above phospholipid fractions were recordable only for variant A (MHS group) as compared to D (MHN), in other words, for the homozygous genotypes, once the above results had been rearranged within MHS and MHN along with different mating variants and genotypes. However, no unambiguous results were obtainable for the heterozygous genotypes of mating variants B and C. Possible underlying reasons are discussed in some detail. The results obtained from mating variants A and D are likely to confirm earlier findings and seem to suggest that the sarcoplasmic reticulum is the primary site of origin of susceptibility to halothane or malignant hyperthermia.

[Animal experiment studies on the healing of bone transplants by clamping plate fixation following hyperthermic denaturation]. / Tierexperimentelle Untersuchungen zur Einheilung von Knochentransplantaten unter Spannplattenfixation nach hyperthermischer Denaturierung.

The re-use of resident bone transplants after hyperthermic denaturalization is demonstrated in radiologically, clinically, and histologically in the lower jaw of the domestic pig. Periosteal and enosteal osteogenic effect is induced.

[Alloxan induced long term diabetes in the domestic pig. I]. / Alloxaninduzierter Langzeitdiabetes beim Hausschwein. I. Mitteilung.

An insulin-deficient diabetes of long duration was induced by intravenous injection of alloxanmonohydrate (200 mg/kg and 100 mg/kg body mass, respectively) in 23 pigs (Landrace, Duroc) of 20-44 kg body mass. The dose of 200 mg/kg body mass was too high and there were fatal outcomes only. With the dose of 100 mg/kg and an initial body mass of 30-35 kg it is possible to induce a chronic diabetes mellitus in the pig. As complications we observed intoxications caused by alloxan, hypoglycemic situations from the 6th hour after application of alloxan with possible hypoglycemic damages, and septic complications by the intravenous catheter. Five out of 23 animals are still alive, 2 animals for 31 months and the others for 19 months. All the living animals have plasma glucose levels between 11 and 17 mmol/1 with very low values of plasma insulin. The serum triglycerides increased slightly in some cases, but not the serum cholesterol. The weight gain is retarded.