Effect of pre-slaughter handling, exercise and the presence of a dog on lamb welfare and meat quality.

Before slaughter, lambs may experience several stressors such as feed and water deprivation, handling and transport that have the potential to negatively impact welfare and meat quality. The objective of this study was to evaluate the effect of pre-slaughter handling, exercise and the presence of a dog on the behaviour and physiology of lambs and meat quality at slaughter. At 6 months of age, 60 lambs (n=20 lambs/replicate; three replicates) were allocated to one of the two treatment groups (n=30 lambs/treatment): low (LOW) intensive handling or high (HIGH) intensive handling. LOW lambs were moved short distances, quietly and without the use of a dog before transport. HIGH lambs were moved quickly, long distances and with a dog present before transport. Lamb behaviour (standing, lying, rumination and panting) was recorded for 1 h before (post-treatment) and after transport (post-transport), and for 30 min before slaughter (pre-slaughter). Blood samples were collected before (baseline), after transport (post-transport) and at exsanguination (at slaughter) to assess cortisol, lactate and non-esterified fatty acid (NEFA) concentrations. At slaughter, lamb carcases (M. longissimus lumborum) were evaluated for pH levels, drip and cook loss, and tenderness. HIGH lambs spent more time standing (P<0.001) and panting (P<0.001) and less time lying (P<0.001) and ruminating (P<0.001) post-treatment than LOW lambs, but more (P<0.001) time ruminating post-transport. All lambs spent more time standing (P<0.001) and less time lying (P<0.001) and panting (P<0.001) post-transport and pre-slaughter than post-treatment. Cortisol concentrations were greater (P<0.001) in lambs post-transport and at slaughter compared with baseline values. Lactate concentrations were lower (P=0.002) in HIGH than LOW lambs. In addition, NEFA concentrations were higher (P<0.001) post-transport and at slaughter in HIGH compared with LOW lambs. Ultimate pH was higher (P<0.001) in HIGH than LOW lambs and pH declined quicker (P=0.012) in LOW than HIGH lambs. Cook loss, drip loss and shear force were lower (P⩽0.05) in HIGH than LOW lambs. The HIGH intensive pre-slaughter handling regime used in the present study caused stress in lambs and increased ultimate pH that could potentially negatively impact welfare, product quality and consistency.

Small heat shock proteins and toughness in intermediate pHu beef.

Bull M. longissimus dorsi (n=94) categorised into high (n=28), intermediate (n=14) and low (n=52) ultimate pH (pHu) were aged at -1.5°C for 28days. Shear force was higher and more variable (p<0.05) in intermediate pHu samples during ageing. Titin, filamin and desmin degradation was also less extensive in intermediate pHu samples compared to the other two pH categories. The extent of the decline of HSP20, HSP27 and αß-crystallin concentrations during post mortem ageing was pHu related such that high pHu meat maintained the highest concentration of small heat shock proteins followed by intermediate and low pHu meat. µ-Calpain autolysis was slowest in intermediate pHu and cathepsin B activities remained consistently low during ageing in this group (p<0.05). Meat toughness in the intermediate pHu group may be attributed to the combination of a larger pool of sHSP with a sub-optimal cathepsin B activity and intermediary µ-calpain activities.

Genetic mapping of quantitative trait loci for meat quality and muscle metabolic traits in cattle.

A whole-genome scan was carried out in New Zealand and Australia to detect quantitative trait loci (QTL) for live animal and carcass composition traits and meat quality attributes in cattle. Backcross calves (385 heifers and 398 steers) were generated, with Jersey and Limousin backgrounds. The New Zealand cattle were reared and finished on pasture, whilst Australian cattle were reared on grass and finished on grain for at least 180 days. This paper reports on meat quality traits (tenderness measured as shear force at 4-5 ages on two muscles as well as associated traits of meat colour, pH and cooking loss) and a number of metabolic traits. For meat quality traits, 18 significant QTL (P < 0.05), located in nine linkage groups, were detected on a genome-wise basis, in combined-sire (seven QTL) or within-sire analyses (11 QTL). For metabolic traits, 11 significant QTL (P < 0.05), located in eight linkage groups, were detected on a genome-wise basis, in combined-sire (five QTL) or within-sire analyses (six QTL). BTA2 and BTA3 had QTL for both metabolic traits and meat quality traits. Six significant QTL for meat quality and metabolic traits were found at the proximal end of chromosome 2. BTA2 and BTA29 were the most common chromosomes harbouring QTL for meat quality traits; QTL for improved tenderness were associated with Limousin-derived and Jersey-derived alleles on these two chromosomes, respectively.

Genotypic effects of calpain 1 and calpastatin on the tenderness of cooked M. longissimus dorsi steaks from Jersey x Limousin, Angus and Hereford-cross cattle.

Single nucleotide polymorphisms (SNPs) in the calpain 1 (CAPN1) and calpastatin (CAST) genes were studied to determine their effects on meat tenderness in Bos taurus cattle. Strip loins (M. longissimus dorsi) were removed from cattle in four resource populations after slaughter (n = 1042), aged under controlled conditions until fixed times after rigor mortis, cooked and measured using a tenderometer. Animals were genotyped for the CAPN1 SNP c.947C>G (p.Ala316Gly; AF252504) and for the CAST SNP c.2959A>G (AF159246). Frequencies of CAPN1 C alleles ranged from 23% to 68%, and CAST A alleles from 84% to 99.5%. From all data combined, the CAPN1 CC genotype (compared with the GG genotype) was associated with a 20.1 +/- 1.7% reduced average shear force at intermediate stages of ageing (P < 0.001) and with a 9.5 +/- 1.3% reduction near ultimate tenderness (P < 0.001). The heterozygote was intermediate. For CAST, corresponding values for AA compared with AG genotypes were reductions of 8.6 +/- 2.0% and 5.1 +/- 1.6% respectively (both P < 0.001), but there were too few GG genotypes for comparison. There were small interactions between the CAPN1 and CAST genotypes. For the CAPN1 and CAST genotypes combined, the maximal genotype effect in average shear force was 25.7 +/- 5.5% (P < 0.001) at intermediate stages and 15.2 +/- 4.8% near ultimate tenderness (P < 0.01).

Post-mortem calpain-system kinetics in lamb: Effects of clenbuterol and preslaughter exercise.

The effect of dietary supplementation with clenbuterol for either 8 days or 55 days in lambs was studied. The 55-day treatment was combined with an immediate preslaughter exercise regime. The effect of these treatments on post-mortem calpain system activities, meat ageing and meat quality was studied. Neither short-nor long-term supplementation had an effect on the rate of pH fall post mortem. Short-term supplementation had no effect on the initial nor the final shear force values but these were higher at intermediate times. In contrast, prolonged supplementation increased shear force values at all times post mortem. Preslaughter exercise, while influencing the rate of pH decrease in both control and supplemented groups, did not affect meat tenderness. After short-term clenbuterol-supplementation, the in-vitro µ-calpain activity was significantly lower in the supplemented group at 6 and 24 hr post mortem, while m-calpain and calpastatin activities were largely unaffected. In contrast, 55-day clenbuterol supplementation resulted in significantly higher levels of calpastatin activity at all times post mortem. These data imply that clenbuterol results in toughened meat due to alterations in the calpain/calpastatin system, the mechanisms of which are dependent upon the duration of supplementation.

GH regulation of the Type 2 IGF receptor in regenerating skeletal muscle of rats.

GH enhances skeletal muscle growth, and IGF-II peptide is highly expressed during regeneration. We have therefore investigated the effect of GH administration on IGF-II binding and expression in regenerating rat skeletal muscle using the techniques of receptor autoradiography and in situ hybridisation. Notexin, a myotoxin, was injected into the right M. biceps femoris (day 0), causing affected fibres to undergo necrosis followed by rapid regeneration. Animals were administered either GH (200 micrograms/100 g body weight) or saline vehicle daily. Contralateral muscles were used as regeneration controls. GH administration during regeneration resulted in significant increases in body weight, and damaged and undamaged muscle weights (P < 0.001). IGF-II expression, which was examined in regenerating fibres, survivor fibres and undamaged fibres, varied according to tissue type (P < 0.001). Specifically, IGF-II expression in regenerating fibres was elevated relative to control and survivor fibres after day 3 (P < 0.05), with a peak on day 9 (P < 0.001). GH did not affect IGF-II message levels. 125I-IGF-II binding in regenerating muscle was examined in the same fibre types as well as in connective tissue. 125I-IGF-II binding in regenerating fibres was higher (P < 0.001) than in other tissue types on day 5. GH administration increased 125I-IGF-II binding in all damaged muscle tissues on day 5 (P < 0.001, regenerating fibres; P < 0.01, others). We believe that this shows for the first time an effect of GH on the Type 2 IGF receptor in regenerating skeletal muscle.

Post-mortem kinetics of meat tenderness and the components of the calpain system in bull skeletal muscle.

Eight strip loins (M. longissimus dorsi) from pasture fed Friesian bulls were aged at 15 °C for a range of times from 1 to 120 h. pH declined from 6.29 (SE 0.119) one hour post slaughter to an ultimate pH of 5.48 (SE 0.013). The activities of the components of the calpain system (µ-calpain, m-calpain and calpastatin) were determined after separation on a DEAE-sephacel column. There was a dramatic decline in µ-calpain activity post slaughter with a complete disappearance within 48 h. The rates of decline in m-calpain and calpastatin activity were slower with 30% and 50% remaining 120 h post slaughter, respectively. The rapid decline in µ-calpain activity relative to the calpastatin activity is likely to reduce the degree of tenderisation and ultimate tenderness of the meat.

Alteration of maternal growth hormone levels during pregnancy influences both fetal and postnatal growth in rats.

To study the effects of exogenous growth hormone (GH) administration to the mother on fetal growth, dwarf rats with an isolated GH deficiency were given daily s.c. injections of GH throughout pregnancy. Fetuses were heavier in GH-treated mothers (p < 0.001), and pups from GH-treated mothers continued to grow faster (p < 0.001) than pups from control mothers throughout the postnatal growth period through to weaning. In normal Wistar rats, administration of a potent antiserum to somatostatin to pregnant rats increased the mean birth weight of the offspring (p < 0.01). Administration of GH to the mothers also significantly increased birth weight (p < 0.05), but administration of antiserum to rat GH resulted in a significant retardation of both fetal and placental growth (p < 0.001). These data suggest that maternal GH status is significantly involved in the growth of the fetal rat.

Evidence of a role for growth hormone, but not for insulin-like growth factors-I or -II in the growth of the neonatal rat.

Neonatal rats were injected with antiserum raised against either insulin-like growth factor (IGF)-I, IGF-II, rat growth hormone (rGH) or somatostatin (SRIF) on each of days 2-5 of life: controls received normal sheep immunoglobulin. Plasma levels of rGH and IGF-I were measured by radioimmunoassay and growth rates recorded. Neonatal administration of anti-rGH resulted in the suppression of plasma IGF-I levels at day 21 and of body weight gain compared with control animals from day 5 of age; relative growth velocity continued to diverge in the absence of any further treatment. Immunoneutralization of IGF-I or of IGF-II had no effect on growth rates of rats at any time during the experiment and had no effect upon plasma rGH concentrations at day 21. However, at day 7, plasma rGH was lower in anti-IGF-I-treated rats than in controls; in contrast, plasma rGH in anti-IGF-II-treated animals at day 7 was higher than in controls. Plasma levels of IGF-I at 49 days of age were similar regardless of the neonatal immunization treatment received. Anti-SRIF treatment of neonatal rats was associated with elevated rGH levels, but no significant stimulation of growth. These results indicated that growth hormone, but not circulating IGF-I or IGF-II are essential for normal growth in the neonatal rat.

Anabolic effects of insulin in growing lambs.

Romney cross-bred wether lambs (aged 16-56 weeks and weighing 16-36 kg) were fed a concentrate diet ad lib. and surgically prepared with catheters in the left and right external iliac arteries and veins, inserted via the ipsilateral medial saphenous artery and vein. Sterile saline was continuously infused through all catheters and after appetite had fully recovered, bovine insulin was infused into one arterial catheter at rates of 5, 10 or 20 mU min-1 for periods of 15, 30 or 45 days in different lambs. At the end of the infusion the lambs were slaughtered to measure the composition of each hindquarter by tissue dissection and chemical analysis. As a result of the insulin infusions, plasma insulin concentrations were increased in venous blood sampled downstream from the infusion site and the arteriovenous concentration differences of glucose were greater across the treated than the contralateral limb. At the 10 and 20 mU min-1 infusion rates, systemic plasma insulin concentrations were increased and glucose concentrations decreased. Pooling results from all the treated lambs showed that intra-arterial insulin infusion increased weight of the hindquarter by 7%, weight of muscle by 5%, weight of dissected fat by 12% and weight of chemical fat by 11% when compared with the contralateral limb. No significant differences were detected among infusion rates or times of infusion. Much of the variation in response between lambs was negatively correlated to their age and/or liveweight when they entered the study. The results demonstrate that tissues in the hindlimb respond to local concentrations of a circulating hormone, provide strong evidence that insulin is an anabolic hormone in the growing postnatal lamb and suggest that such responses diminish with age.