Effects of the halothane genotype and slaughter weight on texture of pork

The objective of this study was to investigate the effects of the halothane (HAL) genotype, slaughter weight (SW), and the HAL x SW interaction on compositional and textural traits of raw and cooked pork. Pigs were bred to exhibit one of the three HAL genotypes (NN, Nn, and nn) with otherwise equivalent genomes. The nn halothane reactors are known to typically produce PSE pork, whereas NN pigs do not typically produce PSE pork. Pietrain x Large White gilts and boars, all with verified Nn genotype (by DNA test), were mated to obtain F2 littermates of the three HAL genotypes. These pigs were slaughtered at either 101 +/- 3 ("light") or 127 +/- 3 ("heavy") kg BW and were evaluated for longissimus muscle traits. The pH at .5 h after death (pH1) was 6.35, 6.13, and 5.68 in NN, Nn, and nn pigs, respectively. Sarcomere length was greater in nn than in NN and Nn pigs (1.94 vs 1.83 and 1.85 microm, respectively). Mechanical resistance was higher in nn than in NN pigs for both raw and cooked meat. Meat from nn pigs was judged by a trained panel to be less rough, more cohesive, harder, more fibrous, less granular, more elastic, and less easy to swallow than meat from NN pigs. For most traits under study, the heterozygotes were intermediate between the homozygotes but closer to NN than to nn pigs. Muscle from heavy pigs had longer sarcomeres and less moisture than muscle from light pigs. The n allele of the HAL gene unfavorably affects pork texture, and this effect is maintained throughout the range of 101 to 127 kg BW.     

猪氟烷基因的检测及其对生长与肉质的影响

对中国瘦肉猪新品系ＤＩ和ＳＪ系３０１头仔猪进行氟烷测验,阳性率为１．３３％。随机抽取氟烷阴性猪３４头进行基因型检测,有６头为杂合子,由此推知ＮＮ、Ｎｎ和ｎｎ的频率分别为８１．２６％１７．４１％和１．３３％,Ｎ和ｎ的频率分别为８９．９７和１０．０３％。对三种基因型的肉质进行评定结果显示,４头氟烷阳性猪（ｎｎ）肉质最差,出现３头严重ＰＳＥ肉和一头轻度ＰＳＥ肉,而氟烷阴性猪未发生ＰＳＥ肉。２８头ＳＪ系氟烷阴性猪用阳性公猪测交,对产出的２０窝２０８头后代进行氟烷测定,共检出４窝中的２１头阳性猪,对测交后代进行肥育性能和肉质测定,结果阳性猪的日增重和饲料转化率等较低,屠宰率、眼肌面积和瘦肉率等较高,肉质较差。     

苏钟Ⅰ系和Ⅱ系猪的氟烷基因及其对产肉性能的影响

:对 3 0 1头瘦肉型新品系苏钟 系和 系仔猪进行氟烷测验 ,阳性率为 1 .3 3 %。随机抽取氟烷阴性猪 3 4头进行基因型检测 ,有 6头为杂合子 ,由此得知 NN、Nn和 nn3种基因型的频率分别为 81 .2 6%、1 7.4 1 %和 1 .3 3 %,N和 n2种基因的频率分别为 89.97%和 1 0 .0 3 %。对 3种基因型猪的肉质进行评定结果显示 ,4头氟烷阳性猪 ( nn)肉质最差 ,出现 3头严重 PSE肉和 1头轻度 PSE肉 ,而氟烷阴性猪 ( NN或 Nn)未发生 PSE肉。2 0头苏钟 系氟烷阴性猪用阳性公猪测交 ,对产出的 2 0 8头后代进行氟烷测验 ,共检出 4窝中的 2 1头阳性猪。对测交后代进行肥育性能和肉质测定 ,发现阳性猪的日增重和饲料报酬等较低 ,屠宰率、眼肌面积和瘦肉率较高 ,肉质较差     

The effect of cyclopiazonic acid on the development of pale, soft, and exudative pork from pigs of defined malignant hyperthermia genotype

Malignant hyperthermia (MH) and the mycotoxin cyclopiazonic acid (CPA) are each associated with abnormal calcium homeostasis in skeletal muscle, a key underlying factor in the development of pale, soft, and exudative (PSE) pork. To determine whether the natural presence of CPA in livestock feed ingredients contributes to the varying incidence of PSE in the pork industry, various levels of CPA (.1 to 50 mg/kg of diet) were included in the diets of market weight hogs (n = 52) of defined malignant hyperthermia genotype (NN = normal, Nn = a MH carrier, and nn = MH-positive). Animals with two copies of the MH mutation (nn) displayed improved live animal performance compared with NN and Nn animals (increased feed intake, average daily gain, and feed efficiency) but yielded lower quality loin chops as indicated by lower 45-min pH (P<.01), higher Commission Internationale de l'Eclairage (CIE) L* color coordinate values (P<.05), and higher drip losses (P<.01). The effects of CPA varied. In the first feeding trial, conducted under normal outside temperatures (2 degrees C), CPA had no effect (P> .2) on either live animal performance or meat quality. During the second trial, conducted under extreme outside temperatures (-18 degrees C), CPA-dependent reductions (P<.05) in feed intake, average daily gain, and 45-min pH in nn hogs support the possibility of interactions between malignant hyperthermia and dietary CPA on skeletal muscle calcium homeostasis and the development of PSE pork. These results suggest that this interaction may require stressful environmental conditions or the ingestion of CPA doses much higher than occur under natural conditions.     

Influence of transport stress on serum cortisol and thyroid hormones in pigs with Halothane gene

The influence of transport stress on rectal temperature, respiratory rate and serum concentrations of cortisol, thyroxine and triiodothyronine (T3) in normal pigs (NN) and in heterozygous (Nn) pigs with Halothane gene (n), a major gene that causes the pale, soft, exudative meat (PSE), were examined in a hot environment (32–34°C). Easily releasable myofilament content of Longissimus dorsi taken by needle biopsy was also measured as an index of myofibrillar disassembly. Rectal temperature and respiratory rate were significantly increased by the transport. However, the influence was higher (P < 0.01) in Nn pigs. Serum concentration of cortisol was increased (P < 0.01) by the transport despite the genotypes. Serum concentration of T3 was not changed by the transport in Nn pigs, whereas in NN pigs it was significantly decreased after the transport (P < 0.01). Easily releasable myofilament content after the transport was higher in Nn pigs than that of NN pigs (P < 0.05). These results suggest that response of thyroid function might relate to the stimulation of muscle proteolysis and PSE in Nn pigs under stressful conditions.     

Restraint effects on stress-related hormones and blood natural killer cell cytotoxicity in pigs with a mutated ryanodine receptor

A mutation in the ryanodine receptor gene (RYR1) of the calcium release channel is responsible for increased stress susceptibility in pigs. In the present study, the relation of a mutation in RYR1 with the neuroendocrine (stress-related hormone) response and the immune defense represented by natural killer cell cytotoxicity (NKCC) during a 4-h restraint and recovery phase in 60 male pigs was investigated. Blood samples were collected from pigs previously divided into RYR1 genotypes (nn, Nn, NN), based on PCR amplification and restriction analyses. The blood samples collected during the restraint and recovery phases of the experiment were used to determine NKCC (⁵¹Cr-release assay), large granular lymphocyte number (hematologic method), and plasma concentrations of prolactin (PRL), GH, ACTH, and cortisol (COR) (by specific RIA). The greatest degree of NKCC response (P < 0.05) to restraint stress relative to controls was observed for the stress-susceptible homozygote group (nn). Measures of stress-related hormones were positively correlated with NKCC during the entire experimental period (P < 0.001 for all investigated hormones) in the nn group. Immunostimulatory effects in the early (0–60 min) phase of restraint were associated with increased hormone responses, especially PRL and GH. In the late (180–240 min) phase of stress and the recovery phase (480 min), a decrease in immune response was accompanied by an elevated COR response in all RYR1 genotypes. Moreover, divergent responses of both PRL (greatest in nn, P < 0.001) and GH (greatest in NN, P < 0.001) to the 4-h restraint were observed. Our results suggest that stress-susceptible RYR1-mutated homozygotes develop a greater level of immune defense, including cytotoxic activity of NK cells, and accompanied by more pronounced stress-induced changes in neuroendocrine response than stress-resistant heterozygous (Nn) and homozygous (NN) pigs.     

Creatine monohydrate supplementation and the quality of fresh pork in normal and halothane carrier pigs

The objective of this research was to examine the impact of supplementation with creatine monohydrate (CMH) on the quality of various muscles from normal and heterozygous halothane carrier pigs. Twenty-nine crossbred pigs, 16 normal (NN) and 13 halothane carrier (Nn) genotypes, were supplemented with 0 or 25 g x pig(-1) x d(-1) of CMH for 5 d before slaughter. Supplemented pigs gained 2.26 kg more weight (P < 0.05) during 5 d of supplementation. There were trends (P < 0.10) toward higher objective marbling scores and lower cooking loss for supplemented pigs. The 45-min pH was 0.27 units higher (P < 0.05) for supplemented pigs in the semimembranosus; CMH supplementation did not influence (P > 0.05) drip loss or muscle composition. Supplementation with CMH also resulted in lower L* values in two ham muscles, semitendinosus (5.15 units) (P < 0.05) and semimembranosus (1.95 units) (P < 0.10) for Nn carcasses. Genotype had significant effects on most quality indicators, with Nn carcasses producing lower-quality lean as evidenced by less desirable subjective and objective color and higher drip losses. Genotype also affected the composition of several muscles, with the NN carcasses having more fat and less moisture.     

Effects of transport stress on concentrations of cortisol, corticosteroid-binding globulin and glucocorticoid receptors in pigs with different halothane genotypes

Effects of stress on concentrations of cortisol and corticosteroid-binding globulin (CBG) in blood plasma and on glucocorticoid receptor concentrations in muscle cytosol were studied in pigs representing three Halothane (Hal) genotypes (NN, Nn, nn). At 12 wk of age, animals were divided into four groups: pigs subjected to transport (5 h in a truck), pigs treated with amperozide prior to transport, pigs not transported but treated with amperozide and pigs neither transported nor given amperozide. Animals were slaughtered the week they reached 100 kg live weight (3 mo later). The Hal gene showed no major influence on the variables studied except for cortisol concentrations (P = .06) measured directly after transport at 12 wk of age (NN = 66.8 nM, Nn = 61.4 nM, nn = 69.4 nM). However, the response in each Hal genotype differed, depending on whether or not the pigs had been exposed to transport. Two weeks after transport, NN pigs developed higher cortisol concentrations than untransported animals, whereas the response was reversed in nn animals; Nn pigs showed no difference in this regard. At slaughter, the effect of transport (12 wk of age) on cortisol and CBG was still apparent. In NN pigs cortisol and CBG concentrations were elevated (P less than or equal to .05, P = .08, respectively), whereas concentrations tended to be lower in nn pigs (P = .17, P = .07, respectively) when compared with untransported pigs. Transported pigs had lower receptor concentrations at slaughter (P less than or equal to .01) than untransported pigs. However, pigs given amperozide in connection with transport had a receptor concentration comparable to that in untransported pigs. Our study shows conclusively that transport stress had long-term effects on cortisol, CBG and glucocorticoid receptor concentrations. In addition, amperozide had long-term effects on cortisol and receptor concentrations.     

Altered calpain levels in longissimus muscle from normal pigs and heterozygotes with the ryanodine receptor mutation

The calpain proteolytic system was examined in the longissimus muscle (LD) of heterozygote pigs carrying a single copy of a mutation in the skeletal muscle ryanodine receptor gene (RyR1) that is associated with porcine stress syndrome and reduced meat quality. Conventional British White-type pigs (n = 30) were selected from a commercial line on the basis of slaughter weight, backfat depth, and pH at 45 min postmortem > 6.0; based on DNA analysis, 11 were heterozygous RyR1 mutants (Nn), and 19 were normal genotype (NN). The LD samples were taken from carcasses at 2, 4, and 24 h postmortem for calpain analysis with enzyme assay and immunoblotting, using specific antisera raised against recombinant polypeptides derived from calpain large subunits and calpastatin. Shear force (SF) was measured after conditioning for 8 d at 2 degrees C and did not differ between Nn and NN groups. The extractable activity of mu-calpain decreased over 24 h postmortem (P < .001), with no significant difference in activity between NN and Nn animals at any time. The activity of m-calpain also decreased with time (P < .001), but it was lower at all times in Nn than in normal genotypes (P < .001). After Western blotting, the immunoreactivity of mu- and m-calpain large subunit bands declined over 24 h postmortem (P < .001); values for mu-calpain were higher (P < .05) and for m-calpain were lower (P < .001) in heterozygotes than in normal animals at each sampling time. The calpastatin antibody detected a major band of 135 kDa that declined with time postmortem but did not differ between Nn and NN genotypes at any sampling time. These data indicate that the levels of extractable mu- and m-calpain, but not calpastatin, may be different in pigs that carry the RyR1 mutation.     

Giant muscle fibres in pigs with different Ryr1 genotype

This study examined the frequency, morphological and immunohistochemical characteristics of the giant fibres in the longissimus muscle of local Krsko polje pigs with different Ryr1 genotypes. Giant fibres were round-shaped and had significantly increased cross-sectional area compared with normal muscle fibres. Only fast-twitch glycolytic fibres were affected, usually showing enhanced succinate dehydrogenase activity. On the ultrastructural level, the dilation of the sarcoplasmic reticulum, swelling of mitochondria and destruction of myofilaments was observed. The incidence of giant fibres was the highest in Ryr1 dimutant pigs (Ryr1 nn), which also exhibited lower muscle pH1 than heterozygous (Ryr1 Nn) or pigs with the wild Ryr1 gene (Ryr1 NN). However, the giant fibres were also present in pigs free of Ryr1 gene mutation. Our results suggest that the giant fibre syndrome depends mostly upon the rate and intensity of early post-mortem glycolysis, which results in acidity of muscle tissue. We suppose that the giant fibre formation is a result of excessive intracellular lactate accumulation in some fast-twitch glycolytic fibres. This process could also explain the ultrastructural alterations and the consequent changes in the oxidative enzymes and myofibrillar ATPase staining pattern observed in our and some previous studies.     

Plasma pyruvate kinase activity vs creatine kinase activity as an indicator of the porcine stress syndrome

Plasma pyruvate kinase (PK) and creatine kinase (CK) activities were increased significantly (P less than 0.001 and P less than 0.05, respectively) in homozygote halothane-reacting pigs (nn), compared with those activities in homozygote nonreacting pigs (NN). Pyruvate kinase activity was less variable within groups than was CK activity, allowing more effective discrimination between nn and NN geno-types. The PK and CK activities in plasma increase with age in halothane-reacting pigs and the nonreacting pigs. Enzyme activities in heterozygote (Nn) nonreacting pigs did not differ significantly (P greater than 0.05) from enzyme activities of homozygote (NN) nonreacting pigs. Although PK was better than CK in identifying stress-susceptible pigs, age-related effects and the failure to identify heterozygotes may restrict the use of plasma PK activity as a diagnostic test for the stress syndrome.     

Impact of RYR1 genotype of Piétrain boars on litter traits of Czech Large White × Czech Landrace crossbred sows

Summary Piétrain boars of different ryanodine receptor (RYR1) genotypes (NN, Nn and nn, three boars each) were mated with approximately 10 Czech Large White × Czech Landrace sows (genotype NN) each to produce one litter per sow. The progeny of nn boars had a significantly higher individual weaning weight (7.31 kg versus 6.86 kg) and average daily gain from birth to weaning (252 g/day versus 240 g/day) than the progeny of NN boars. Furthermore, piglets from nn boars differed statistically significant also from piglets of boars with the Nn genotype for these two traits. The litters of the NN boars were greater by 0.5 piglets than the litters of the nn boars for the total number of piglets born, piglets born alive and piglets weaned, but the differences were not significant. The Nn genotype showed for all litter size traits and the number of stillborn piglets the best values. The difference between the genotypes Nn and nn was statistically significant at the 0.05 level for the number of piglets born alive and the number of piglets weaned.     

A within-breed comparison of RYR1 pig genotypes for performance, feeding behaviour, and carcass, meat and fat quality traits

Data on 187 DNA‐tested purebred Landrace pigs were used to compare heterozygous RYR1 genotypes (Nn) and normal homozygotes (NN) in performance (growth, fat deposition rate and feeding behaviour patterns), and carcass, meat and fat quality traits. From 75 to 165 days of age, live body weight (BW), ultrasonic midback (UMB) and loin (ULB) backfat measurements were recorded periodically on the same animal. Individual voluntary feed intake (DFI), number of visits (NVD) and feeding time (FTD) were measured on a daily basis using an automatic feeding system. Polynomial models with random regression coefficients were used to describe BW, UMB, ULB, DFI, NVD and FTD as a function of age. Carcass, meat and fat quality traits were analysed using a mixed model with the RYR1 genotype as a fixed effect. No significant (p < 0.05) differences were observed between NN and Nn genotypes for growth, feed intake and feed efficiency at any age, but the NN pigs showed higher values for UMB and ULB (0.68 ± 0.30 mm and 0.88 ± 0.40 mm, respectively, at 165 days). The Nn pigs had higher predicted carcass lean content (+11.1 ± 3.7 g/kg) and higher proportion of ham in the carcass (+2.9 ± 1.4 g/kg). However, they showed lower values of pH measurements at 45 min post‐mortem (–0.26 ± 0.05 in M. longissimus dorsi, LD), and higher electrical conductivity at 45 min (+1.03 ± 0.17 μs in LD) and 24 h post‐mortem (+1.63 ± 0.29 μs in LD). No significant differences were found between genotypes for intramuscular fat in the M. semimembranosus (SM) and for fatty acid profiles in the subcutaneous backfat and in the intramuscular fat of the SM. The results of this study suggest that Nn pigs would attain their maximum growth rate at later stages than the NN without any relevant change in the feeding behaviour pattern. Also they confirm that Nn pigs are leaner, have greater proportion of high‐priced cuts in the carcass and are more prone to develop pale, soft and exudative meats. However, no effect on fat quality traits was found.     

Could the pale, soft, and exudative condition be explained by distinctive histological characteristics?

Pork quality depends on various genetic and environmental factors. Despite the improvement of slaughter conditions, the PSE type is still one of the main concerns in this field. This study was conducted on nonstressed animals to evaluate the tissue characteristics of some muscles usually involved during stress compared with a reference muscle, the M. triceps brachii, which is actually not subject to stress-caused damages. Samples of M. triceps brachii, M. longissimus dorsi, M. biceps femoris, and M. semimembranosus were taken from pigs exhibiting 1 of the 3 HAL genotypes (NN, Nn, or nn) and 2 of the 3 RN genotypes (rn+rn+ or rn+RN-). Histoenzymology and immunohistochemistry were used to compare the fiber typing and capillary network in these muscles within these different stress susceptibility genotypes. In comparison with the reference muscle, M. triceps brachii, the combination of a high value of the number of type IIb fibers and a low vascular network showed a primary effect on muscles usually involved during stress. This led to the definition of a PSE index. A dramatic increase (P < 0.001) in this PSE index was systematically found in muscles usually involved in the PSE-type condition. These results show that distinctive histological characteristics were associated with the vulnerability of some muscles independently of the genotypes. Moreover, this study highlights the distinctive histological features of each genotype and is likely to suggest some interactions between them.     

Effects of trailer design on animal welfare parameters and carcass and meat quality of three Pietrain crosses being transported over a short distance

The objective of this study was to evaluate the effects of trailer design on the stress responses and meat quality traits of 3 different pig crosses: 50% Pietrain breeding with HAL^Nn (50Nn), 50% Pietrain breeding with HAL^NN (50NN) and 25% Pietrain breeding with HAL^NN genotype (25NN). Market barrows (n=360), as a subset of 12 trailer loads of pigs, were transported from farm to slaughter on 6 dates in 2009 for 45 min in either a pot-belly (PB) or flat-deck (FD) trailer, with 120 pigs/genetic group being represented. Temperature (T) and relative humidity (RH) were recorded by data loggers mounted in both trailers. Behaviours and handler interventions were video-taped at loading, unloading and in lairage. At exsanguination, blood samples were collected for the assessment of lactate, cortisol, creatine kinase (CK), haptoglobin and Pig-MAP concentrations. Meat quality was measured in the longissimus dorsi (LD), semimembranosus (SM) and adductor (AD) muscles of all pigs. Temperatures were warmer in compartments 6 and 11 at loading (P<0.001), compartment 11 during travelling (P=0.05), and in compartment 5 at unloading (P=0.01) of the PB trailer. Pigs unloaded from the FD trailer overlapped more (P<0.001), whereas (P<0.001) the frequency of jamming was noted for pigs unloaded from the PB trailer. Pigs with 50% Pietrain genetics overlapped and jammed more (P<0.001) than pigs with 25% Pietrain genetics, regardless of HAL status. Greater (P=0.03) CK levels were found in 50Nn pigs transported in the PB trailer, while 50Nn and 50NN pigs had greater (P=0.028) lactate levels than 25NN pigs. Carcasses from 50Nn and 50NN pigs were leaner (P=0.04), and the skin damage score was lower (P=0.04) in 25NN carcasses. Overlap-type bruises were greater (P=0.02) in pigs transported in the FD trailer. Pigs transported in the PB trailer had greater (P=0.05) pHu in the SM and AD muscles (P=0.013). Except for pHu in the SM muscle, all meat quality parameters were affected by the Hal gene (P=0.04). The use of a PB trailer for short distance transportation of pigs to slaughter negatively affected stress responses and meat quality. The greater proportion of Pietrain genetics in the selection resulted in leaner carcasses, but also in pigs being more difficult to handle. Crossbreeding appeared to have a greater impact on animal welfare and meat quality than vehicle type, but trailer type may emphasize these negative genotype-related defects.     

The effect of the Halothane and Rendement Napole genes on carcass and meat quality characteristics of pigs

The objective of this study was to determine the effects of the Halothane (N) and Rendement Napole (RN) genes on carcass and meat quality characteristics in pigs. Halothane and RN carrier (Nn/RN- rn+) Hampshire boars (n = 4) were mated to dams that were homozygous for the normal allele of both genes (NN/rn+ rn+) to produce progeny of four genotypes: 1, NN/rn+ rn+ (n = 31); 2, Nn/rn+ rn+ (n = 27); 3, NN/RN- rn+) (n = 30); and 4, Nn/RN- rn+ (n = 23). A DNA test was used to determine Halothane genotype, and longissimus glycolytic potential was used to predict the RN genotype. Pigs were reared under standard conditions to approximately 120 kg live weight and slaughtered at a commercial plant, and carcass characteristics and meat quality were evaluated. Halothane carriers (Nn/ _ _), in comparison to Halothane normal (NN/_ _) pigs, had shorter carcasses, lower longissimus ultimate pH, higher Minolta L* and b* values, and greater drip loss. Rendement Napole gene carriers (_ _/RN- rn+) had higher L* and b* values and drip and cooking loss and lower longissimus ultimate pH than homozygous recessive animals (_ _/rn+ rn+). There were Halothane x RN genotype interactions (P < 0.05) for subjective color, firmness, and marbling scores, and for shear force. Animals that were normal for both genes (NN/rn+ rn+) had the highest subjective scores for color (2.60, 1.88, 1.85, and 1.95, SE = 0.181, P < 0.05), firmness (2.53, 2.03, 2.10, and 1.89, SE = 0.182, P < 0 .05), and marbling (2.11, 1.44, 1.53, and 1.55, SE = 0.153, P < 0 .05) for genotypes 1, 2, 3, and 4, respectively, suggesting darker, firmer muscle with a higher level of marbling for this genotype. Shear force was highest for Nn/rn+ rn+ animals (3.83, 4.41, 3.79, and 3.70, respectively, SE = 0.172, P < 0.05). Gilts had less s.c. backfat thickness, greater longissimus muscle area, and lower subjective marbling scores than barrows. There was no effect (P > 0.05) of gender on other meat quality traits. This study illustrates the negative effects of the Halothane and RN genes on fresh pork quality and suggests that in combination the detrimental effects of the two genes are additive for ultimate pH, objective color, and water-holding capacity.     

Serum pyruvate kinase and its relation to stress susceptibility in pigs

SUMMARY: Four blood tests, complementary to the halothane test, have been screened for their capacity to discriminate between homozygous and heterozygous stress resistant and stress susceptible pigs: TBARS in plasma, lipid peroxides from erythrocytes and plasma creatine and pyruvate kinase activity. Blood samples were collected from homozygous (NN) and heterozygous (Nn) stress resistant and homozygous stress susceptible (nn) pigs, while subjected to the halothane test. Serum creatine kinase and pyruvate kinase tests were retained for a more elaborated study. Pyruvate kinase appears to be a more reliable indicator of stress susceptibility than the well-known creatine kinase. However both parameters display a considerable standard deviation. Consecutive assays show the pyruvate kinase activity to be dependent on age (weight), which accounts for part of the variation; other sources of variation are the conditions of the exeriment and the way the imposed stress is perceived by the animal. During this experiment a rapid and accurate DNA-test for HAL-genotype detection has been developed (Fujii et al. 1991; Otsu et al. 1991; Coppieters et al. 1992). In spite of this, the pyruvate kinase parameter is a much more discriminating and remarkable indicator for stress susceptibility than the, in previous years, widely used creatine kinase test. ZUSAMMENFASSUNG: Serum Pyruvat Kinase und seine Relation mit Streβanf?lligkeit bei Schweinen Vier Blutteste, komplement?r dem HAL-Test, wurden in einem Screening-Verfahren untersucht um homozygote und heterozygote stre?resistente und stre?anf?llige Schweine zu differenzieren: TBARS in Plasma, Lipid Peroxyde von Erythrocyten und Plasma Kreatine und Pyruvat Kinase Aktivit?t. W?hrend des Halothantestes wurde Blut entnommen von homozygoten (NN) und heterozygoten (Nn) stre?resistenten und homozygoten stre?anf?lligen Schweinen (nn). Mit Serum Kreatine Kinase und Pyruvat Kinase Aktivit?t Tests wurden eine umfassende überprüfung durchgeführt. Letztere zeigt sich als ein zuverl?ssiger Indikator der Stre?anf?lligkeit als die wohl bekanntere Kreatine Kinase. Mehrmalige Analysen zeigen, da? Pyruvat Kinase Aktivit?t vom Alter (Gewicht) abh?ngig ist, was teilweise die Variabilit?t verursacht; andere Quellen der Variabilit?t sind die experimentellen Umst?nde und die Weise, wie der auferlegte Stre? durch das Tier erfahren wird. W?hrend der Durchführung dieses Experimentes wurde ein schneller und akkurater DNA-Test für HAL-Genotyp Detektion entwickelt (Fujii et al. 1991; Otsu et al. 1991; Coppieters et al. 1992). Im Vergleich mit dem, in vorgehenden Jahren, h?ufig angewendeten Kreatine Kinase Test, ist der Pyruvat Kinase Parameter trotzdem ein bemerkenswerter und diskriminierender Indikator für Stre?anf?lligkeit.     

Genetic polymorphism of the 17th exon at porcine RYR1 locus: a new variant in a local Portuguese pig breed demonstrated by SSCP analysis

Until 1991, malignant hyperthermia represented a major and important economic loss in modern pork production. It is a recessive autosomal disorder with variable expression that was not known to occur in the local pig breeds from Portugal, Alentejano and Bísaro, prior to outcrossing with commercial breeds. In swine, this disorder is due to a mutation in exon 17 of the ryanodine receptor 1 (RYR1) gene, detectable through the analysis of a restriction fragment length polymorphism. We used this technique to distinguish between resistant (NN), carrier (Nn) and susceptible (nn) genotypes, with the aim of excluding carriers from the breeding programme. We further developed a rapid screening technique on the basis of SSCP (single-strand conformation polymorphism). Fifteen heterozygous (Nn) individuals were found in our sample of 48 individuals from Bísaro breed using both techniques. The absence of homozygous susceptible pigs suggests that the introduction of this allele into Bísaro breed has been recent. A genetically controlled breeding programme on the basis of our findings should help to secure the future of Bísaro pigs of which few individuals are known to exist. To date, only one mutation was found in the 5' end of exon 17 of the porcine RYR1 gene. Here we present the frequencies of this mutation and describe a new mutation detected in the Alentejano breed. PCR-SSCP analysis followed by automatic sequencing allowed the detection of this rare transition (C → T) at nucleotide 1851 of exon 17.     

Diminished expression of ACTH signaling proteins and steroidogenic limiting factors in adrenocortical cells isolated from halothane nn pigs

Previous studies demonstrated significantly lower plasma cortisol level in homozygous halothane-positive (Hal nn) pigs, as compared with homozygous halothane-negative (Hal NN) pigs. To determine whether such difference is attributed to the fundamental alterations in adrenocortical function, F1 offsprings from Pietrain (Hal nn)xErhualian (Hal NN) were intercrossed to produce F2 sibling pigs with segregated genotypes. Adrenocortical cells were isolated from the Hal nn and Hal NN F2 pigs, respectively, and cultured with or without ACTH challenge. Cortisol levels in culture medium, as well as the content of MC2R, cAMP, CREB, phosphorylated CREB (pCREB), StAR and P450scc in adrenocortical cell lysates, were determined. Cortisol, cAMP, StAR and P450scc levels were significantly lower in Hal nn adrenocortical cells under basal condition without ACTH challenge. ACTH significantly increased cortisol level in the medium and the protein content of MC2R, StAR, P450scc in adrenocortical cell lysates, regardless of genotypes. Total CREB protein content was not different between genotypes and treatments, whereas pCREB content exhibited significant effects of genotype and treatment, being higher in Hal NN than in Hal nn under basal condition and in response to ACTH challenge. These results indicate that the compromised cAMP/PKA/pCREB signaling pathway of ACTH and diminished expression of limiting factors in adrenocortical steroidogenesis (StAR and P450scc) may contribute to the significantly lower plasma cortisol levels in Hal nn pigs.