Forty-five Duroc (recognized as not susceptible to stress) and 34 Pietrain (susceptible to stress) pigs were subjected to immobilization stress in a prone position for 5, 15, 30 and 60 min. Plasma concentrations of epinephrine (E), norepinephrine (NE), dopamine (DA) and cortisol (C) were determined in response to restraint stress. The concentrations of E, NE and DA were different between the two strains of pigs (some significant interactions); the highest response was seen after 5 min of stress. The concentration of plasma C increased with duration of stress and there was a significant interaction between strain of animals and the time of stress. Our data substantiate the use of E, NE, DA and C as indicators of stress in swine as early as 5 min after exposure to the stressor. It is also shown that stress-susceptible Pietrain pigs had higher plasma concentrations of E, NE and DA than Duroc pigs.

The relationship between lysosomal proteolytic enzyme activities involved in skeletal muscle proteolysis of the longissimus lumborum et thoracis muscle (MLLT) of bulls was described. Samples from the same region were obtained post mortem from 7 Piemontese (P) and 54 Black-and-White bulls (B-W) about 18 months old fed ad libitum. The activity of cathepsin D was determined as pepstatin (cathepsin D inhibitor) sensitive activity (PSCatD) towards 1% haemoglobin. Pepstatin-insensitive acid (PIA) and leupeptin-insensitive (thiol proteinase inhibitor) acid (LIA) autolytic activities were measured in the presence of 1 mM Mg⁺⁺. MLLT was also analysed for RNA, DNA and protein content. The data were processed by analysis of variance and differences between sires were tested by the contrast procedure of general linear model. In the examined muscle RNA decreased by 16% in B-W compared to P, CPS by about 14% and FCS by about 39%. DNA content was higher by 64.5% in B-W compared to P bulls (P ≤ 0.01). Some differences were found between P bulls and B-W groups of sires in the percentage of proteins (P ≤ 0.01), CatD and PSCatD (P ≤ 0.01), but the most pronounced differences were determined in PIA and LIA (P ≤ 0.01), and in the percentage of inhibition by pepstatin and leupeptin (P ≤ 0.01) in AAA. In the Black-and-White group of sires the percentage of protein and percentage of inhibition by pepstatin and leupeptin in AAA were lowered by about 10, 17 and 22%, but PSCatD, PIA and LIA were higher by about 23.7, 41 and 57.7%, respectively, compared to Piemontese bulls. The level of aspartic and thiol proteinases was lower in the muscles of B-W compared to Piemontese. The activity was much higher in B-W compared to P. These results indicate the faster turnover of proteins in the groups after Black-and-White sires and higher anabolic increase in degradation in Piemontese bulls.    

No abstract

Summary The distribution and endogenous concentrations of norepinephrine (NE), epinephrine (E), dopamine (DA) as well as DA metabolites 3,4‐dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 5‐hydroxytryptamine (5‐HT) and the main 5‐HT metabolite – 5‐hydroxyindole‐3‐acetic acid (5‐HIAA), were measured by HPLC in 15 brain areas of control and immobilized adult Duroc (D) and Pietrain (P) pigs. The animals were immobilized for 15, 30 and 60 min in the prone position. Control pigs displayed the pattern of regional distribution of brain monoamines similar to that described in other species. Absolute values of NE and E were much higher in the hypothalamus and the preoptic area of pigs than in other species. The concentration of E was seven times higher in the hippocampus posterior in P pigs as compared to D pigs. Also, in most structures DOPAC/DA ratio was relatively high, which points to strains related difference in the turnover of dopamine. In control animals, statistical differences in the bioamine contents were seen between both strains in 25 cases. Epinephrine accounted for more than 12% of the total concentration of both amines in D and 15% in P, this proportion was the highest among the structures compared. The highest concentration of DA and its metabolites – DOPAC and HVA, was revealed in the caudate nucleus and in the nucleus accumbens. Of other structures, the substantia nigra (SN), the septum and the ventral tegmental area (VTA) contained relatively high concentrations of DA, DOPAC and HVA. The 5‐HT level varied from the lowest concentration in the hippocampus posterior, cornu ammonis, nucleus accumbens and the caudate nucleus to the highest concentration in the hypothalamus, the SN, and brain stem areas. This distribution was paralleled by corresponding concentration of 5‐HIAA, being the highest in raphe nuclei (RN). The most conspicuous changes produced by immobilization stress consisted of a substantial decrease in the hypothalamic levels of NE and E in both strains of pig. The level of NE was always higher in D than in P in cornu ammonis, hippocampus posterior, nucleus caudatus, SN, septum, hypothalamus medialis, tectum and olfactory tuberculum. The same was for E in: SN, septum, hypothalamus medialis, and olfactory tubercle. DA, DOPAC and HVA, were changed primarily in SN in P pigs, and in RN in P and D animals. Serotonin (5‐HT) level increased substantially in RN in P pigs. 5‐HIAA level was highly elevated in the RN being always higher in P than in D pigs and in PAG. We suggest that restraint immobilization stress lead to compensatory changes in both brain monoaminergic systems. These changes may affect coping with stress demands mostly in the structures that are proposed to play a major role in the pig responsiveness to acute stress conditions.

A major factor limiting the transgenesis in domestic animals is the inefficiency of maintaining large numbers of recipients carrying nontransgenic foetuses. The objectives of this study were: 1. to determine the influence of green fluorescent protein (GFP) construct injection on the development of bovine embryos, 2. to identify and select the GFP positive bovine embryos, and 3. to determine the rate of mosaicism in transgenic embryos. Cattle oocytes were matured and fertilised in vitro and zygotes were microinjected with pCX-EGFP construct consisting of CMV-IE enhancer, chicken β-actin promoter, cDNA of GFP (EGFP-732 bp) and rabbit β-globin polyadenylation sequences. Embryos from control (64) and microinjected (198) groups were cultured in vitro. After 168 h of culture, morula and blastocysts were observed in 39.06% of control and in 23.23% of injected group. We obtained three GFP positive embryos (1.51% of injected zygotes and 6.52% of morulae/blastocysts). One of them was 100, second 75 and third 25% GFP positive (66.7% of mosaicism). Use of gfp gene reporter to select bovine embryos is useful method to increase transgenic offspring, because GFP marker allows to choice only transgenic embryos and transfer them to recipients.