Validating the salivary testosterone and cortisol concentration measures in response to short high-intensity exercise.

AIM: To validate the testosterone (T) and cortisol (C) concentration measures in saliva in response to short high-intensity exercise. METHODS: Nine healthy males provided matching saliva and plasma samples before and after a 30-second Wingate cycle test. Saliva was assayed for T (Sal-T) and C (Sal-C) concentrations, and plasma for total T and total C, sex hormone-binding globulin, corticosteroid-binding globulin (CBG) and albumin concentrations. The plasma free and bioavailable hormones were calculated. RESULTS: The Sal-T and plasma T correlations were weak to moderate (r=0.57-0.61) when examined between individuals (pooled data for all participants), but these relationships improved (r = 0.71-0.73) within individuals (data for each participant on average). The Sal-C and plasma C correlations were strong both between individuals (r=0.81-0.84) and within individuals (r=0.83-0.84). The peak relative increases in Sal-T (35+/-9%) and Sal-C (63+/-29%) concentrations exceeded the plasma total and/or free hormones, but not the bioavailable hormones. Albumin (10+/-3%) and CBG (16+/-4%) also increased with exercise, along with blood lactate (943+/-119%). CONCLUSION: The Sal-T and Sal-C concentration measures were validated in response to short high-intensity exercise, especially for individuals. The hormonal changes in saliva were also more sensitive to exercise (i.e. greater relative responses) than the plasma total and/or free hormones, potentially arising from changes in the binding proteins and blood lactate. These findings support the use of saliva as a medium for steroid determination in sport.

Pre-slaughter stress arising from on-farm handling and its interactions with electrical stimulation on tenderness of lambs.

The effect of electrical stimulation of lamb carcasses (n=269) or its absence (n=257) on shear force of m. longissimus thoracis et lumborum (LT) was monitored during ageing in pasture-fed merino lambs (n=526). The lambs were slaughtered on four different days allowing durations of between one to 10 days of recovery from pre-slaughter handling (yarding, weighing and crutching) that affected ultimate pH (pH(u)). The right LT was removed 20-40min post-slaughter, tightly-wrapped in cling film (prevents the muscle cross-section increasing and thus minimising shortening) and rapidly cooled to 15°C to enter rigor mortis and age. At 0, 4, 24 and 72h post-slaughter, pH measurements and samples for shear force measurement were taken. Pre-slaughter handling had a significant negative effect on pH(u) and several days recovery were required for pH(u) to reach values associated with optimal meat quality as reflected by pH(u). Lambs with one and three days recovery (no significant difference between them) had a pH(u)>5.7 in 50% of the muscles and 19.4%>pH(u) 5.8. Whereas, in lambs with 8-10 days recovery (no significant difference between them), only 8% had a pH(u)>5.7 and 3.1%>pH(u) 5.8. Within each slaughter day electrically stimulated lambs were always more tender than non-stimulated lambs. For non-stimulated muscles at 72h, shear force values >40N occurred for 11.2% of the muscles: for electrically stimulated muscles at 72h, shear force values >40N occurred for 1.9% of the muscles. The rates of tenderisation were slower for intermediate pH(u) values resulting in higher shear force values at all ageing durations. With ageing at 72h for intermediate pH(u), non-stimulated muscles (n=38) 17.64% were >40N and for stimulated muscles (n=34), 7.9% were >40N.

The relationship between postmortem urinary catecholamines, meat ultimate pH, and shear force in bulls and cows.

We investigated the relationship between stress responsiveness and meat quality in cattle. The cattle were 16-19-month-old Mixed (n=37, 303 kg) or Non-mixed (n=23, 279 kg) Friesian bulls and Friesian cull cows (1.5-7 years) that were classified Unfinished (n=133, 195 kg) or Finished for 3 months (n=34, 252 kg). A portion of m. longissimus lumborum (LL) muscle was obtained from consecutive animals for a single days slaughter of 227 cattle processed through a hot boning plant. Catecholamines were obtained from bladder urine. Ultimate pH, lactate, glycogen, glycolytic potential and representative sarcomere lengths were determined from 20-h muscle samples. Shear force measurements were from meat aged at 15 °C for 20 and 90 h. Urinary noradrenaline was similar between Mixed (14.86 ng/µmol creatinine) and Non-mixed bulls (14.07 ng/µmol creatinine) and Finished cows (15.24 ng/µmol creatinine) and elevated in the Unfinished cows (22.28 ng/µmol creatinine). Urinary adrenaline was higher in Mixed bulls (9.5 ng/µmol creatinine) than Non-mixed bulls (5.7 ng/µmol creatinine) and higher in both Unfinished cows (16.67 ng/µmol creatinine) and Finished cows (14.19 ng/µmol creatinine). For bulls that are growing well on-farm, with a only short period of fasting prior to slaughter, pH(u) is a good predictor of meat tenderness. In this situation, stressors that lower muscle glycogen pre-slaughter can have a significant effect on meat shear force, and individual animals with elevated urinary adrenaline were the most susceptible. However, in the case of cows, adrenaline responses are not necessarily associated with glycogen depletion and pH(u) is less affected than for bulls. Furthermore, the ability of meat to tenderise fully, related to reduced proteolytic turnover, is reduced in nutritionally compromised animals such as the Unfinished cows.

A comparison of catecholamine and cortisol responses of young lambs and calves to painful husbandry procedures.

AIM: To compare the changes in plasma concentrations of noradrenaline, adrenaline and cortisol in lambs after ring castration plus tailing and in calves after dehorning with or without prior local anaesthesia. METHODS: Male lambs were castrated and tailed with rings and calves were dehorned by amputation using a scoop with or without prior local anaesthesia. Blood samples were taken before and after treatment and plasma concentrations of noradrenaline, adrenaline and cortisol were determined. RESULTS: Castration plus tailing of lambs resulted in a rapid increase in noradrenaline concentrations, a lack of an adrenaline response and a marked increase in cortisol concentration. There were similar changes in catecholamine concentrations in calves that were dehorned both with and without local anaesthetic, with adrenaline being elevated within 5 min of treatment and noradrenaline exhibiting a more protracted response. Dehorning caused a marked cortisol increase which was reduced to control concentrations by local anaesthesia for as long as the associated nerve blockade lasted. CONCLUSIONS: The very short-lived adrenaline responses in calves were attributed to dehorning-induced nociceptor input leading to sympathetic stimulation of the adrenal medulla. The longer lasting noradrenaline responses in lambs and calves were thought be due to 'wash-out' of noradrenaline from damaged tissue associated with rings and amputation wounds, respectively.

The effect of nutritional supplements on growth rate, stress responsiveness, muscle glycogen and meat tenderness in pastoral lambs.

Nutritional supplements and a magnesium bolus and were used in lambs in a 2×2 factorial design to investigate the effect on growth, preslaughter stress measurements, muscle glycogen, and meat quality. In total, 64 Perendale lambs were used (32.7±0.53 kg, mean±SEM). Feed supplemented animals received 150 g feed pellets day(-1) in addition to pasture grazing, and this increased growth from 183 to 207 g day(-1). Mean delivery of Mg from the boluses was 0.17 g day(-1) for 28 days, with no effect on growth rates, or any other of the variables measured. Urinary noradrenaline, adrenaline and cortisol did not differ between groups in the immediate pre-slaughter period. The meat ultimate pH was not different between groups and had a mean range of 5.47-5.53. Muscle residual glycogen did not differ between groups and had a mean range of 42-43 mmol kg(-1). Finally there were no differences in shear force values at all ageing times. The final shear force value of 2.5 kg F after 72 h ageing at 15 °C was a low value representing tender meat. These studies indicate that if nutrition is adequate and stress levels are low, there are no differences in meat tenderness of pasture fed lambs compared with those having feed supplements to increase growth rate.

Impact of climate on thermal rhythm in pastoral sheep.

The effects of environmental conditions on temperature rhythms were investigated in ewe lambs at pasture. Two groups of 20 lambs had heart rate (HR), vaginal temperature (T(v)), ear-canal temperature (T(c)) and ear-pinna temperature (T(p)) monitored continuously for 3 days. Climatic conditions were recorded at the same time and Temperature Humidity Index (THI) calculated. One group experienced fine clear weather for the 3 days, the other group experienced 2 days of heavy rain. During periods of fine weather, the daily rhythm for T(v) and T(c) was monophasic. However, heavy rain and a constant THI reduced the amplitude of the recorded temperature rhythms. Daily T(v) and T(c) patterns correlated strongly with THI, with a phase lag of 2 h. Peak T(v) and T(c) were at approximately 17:00 h each day. Mean maximum daily amplitudes were approximately 1.3 degrees C for T(v) and T(c). Mean T(v) was 39.3+/-0.1 and 39.6+/-0.1 for weeks 1 and 2, respectively, while mean T(c) was 38.9+/-0.1 and 39.2+/-0.1. Changes in T(v) and T(c) were closely correlated. We conclude that climate has a major effect on body temperature rhythms.

Responses of the red blood cells from two high-energy-demand teleosts, yellowfin tuna (Thunnus albacares) and skipjack tuna (Katsuwonus pelamis), to catecholamines.

In fishes, catecholamines increase red blood cell intracellular pH through stimulation of a sodium/proton (Na+/H+) antiporter. This response can counteract potential reductions in blood O2 carrying capacity (due to Bohr and Root effects) when plasma pH and intracellular pH decrease during hypoxia, hypercapnia, or following exhaustive exercise. Tuna physiology and behavior dictate exceptionally high rates of O2 delivery to the tissues often under adverse conditions, but especially during recovery from exhaustive exercise when plasma pH may be reduced by as much as 0.4 pH units. We hypothesize that blood O2 transport during periods of metabolic acidosis could be especially critical in tunas and the response of rbc to catecholamines elevated to an extreme. We therefore investigated the in vitro response of red blood cells from yellowfin tuna (Thunnus albacares) and skipjack tuna (Katsuwonus pelamis) to catecholamines. Tuna red blood cells had a typical response to catecholamines, indicated by a rapid decrease in plasma pH. Amiloride reduced the response, whereas 4,4'diisothiocyanatostilbene-2,2'-disulphonic acid enhanced both the decrease in plasma pH and the increase in intracellular pH. Changes in plasma [Na+], [Cl-], and [K+] were consistent with the hypothesis that tuna red blood cells have a Na+/H+ antiporter similar to that described for other teleost red blood cells. Red blood cells from both tuna species were more responsive to noradrenaline than adrenaline. At identical catecholamine concentrations, the decrease in plasma pH was greater in skipjack tuna blood, the more active of the two tuna species. Based on changes in plasma pH, the response of red blood cells to catecholamines from both tuna species was less than that of rainbow trout (Oncorhynchus mykiss) red blood cells, but greater than that of cod (Gadus morhua) red blood cells. Noradrenaline had no measurable influence on the O2 affinity of skipjack tuna blood and only slightly increased the O2 affinity of yellowfin tuna blood. Our results, therefore, do not support our original hypothesis. The catecholamine response of red blood cells from high-energy-demand teleosts (i.e., tunas) is not enhanced compared to other teleosts. There are data on changes in cardio-respiratory function in tunas caused by acute hypoxia and modest increases in activity, but there are no data on the changes in cardio-respiratory function in tunas accompanying the large increases in metabolic rate seen during recovery from exhaustive exercise. However, we conclude that during those instances where high rates of O2 delivery to the tissues are needed, tunas' ability to increase cardiac output, ventilation volume, blood O2 carrying capacity, and effective respiratory (i.e., gill) surface area are probably more important than are the responses of red blood cells to catecholamines. We also use our data to investigate the extent of the Haldane effect and its relationship to blood O2 and CO2 transport in yellowfin tuna. Yellowfin tuna blood shows a large Haldane effect; intracellular pH increases 0.20 units during oxygenation. The largest change in intracellular pH occurs between 40-100% O2 saturation, indicating that yellowfin tuna, like other teleosts, fully exploit the Haldane effect over the normal physiological range of blood O2 saturation.