Effect of Short-term Cooling at –70°C in an Air Cryogenic Sauna on Body Temperature and Lipid Profile of Healthy People

The investigation was aimed at evaluation of level and rate of cutaneous and tympanic temperature drop due to a single short-timed (3 min) cooling in a cryosauna (–70°С), and adaptation-indicative physiological parameters, including blood catecholamines, ACTH, lipoproteins and free fatty acids. The subjects were seven healthy men. Exposure to cold invariably reduced the internal (tympanic) and cutaneous temperature by 1°С and 7°С on average, respectively. Tympanic temperature remained 0.4°С low on the average for more than 20 min after exposure. Cutaneous temperature was 1°С below the norm for an hour after cooling. For one hour after the short-term cold exposure, blood norepinephrine remained increased, and so did the blood concentrations of high-density lipoprotein cholesterol and free fatty acids. These results demonstrate brief adaptive changes following a single exposure at–70°C.     

Ontogenetic shifts in thermal tolerance, selected body temperature and thermal dependence of food assimilation and locomotor performance in a lacertid lizard, Eremias brenchleyi

We used Eremias brenchleyi as a model animal to examine differences in thermal tolerance, selected body temperature, and the thermal dependence of food assimilation and locomotor performance between juvenile and adult lizards. Adults selected higher body temperatures (33.5 vs. 31.7 degrees C) and were able to tolerate a wider range of body temperatures (3.4-43.6 vs. 5.1-40.8 degrees C) than juveniles. Within the body temperature range of 26-38 degrees C, adults overall ate more than juveniles, and food passage rate was faster in adults than juveniles. Apparent digestive coefficient (ADC) and assimilation efficiency (AE) varied among temperature treatments but no clear temperature associated patterns could be discerned for these two variables. At each test temperature ADC and AE were both higher in adults than in juveniles. Sprint speed increased with increase in body temperature at lower body temperatures, but decreased at higher body temperatures. At each test temperature adults ran faster than did juveniles, and the range of body temperatures where lizards maintained 90% of maximum speed differed between adults (27-34 degrees C) and juveniles (29-37 degrees C). Optimal temperatures and thermal sensitivities differed between food assimilation and sprint speed. Our results not only show strong patterns of ontogenetic variation in thermal tolerance, selected body temperature and thermal dependence of food assimilation and locomotor performance in E. brenchleyi, but also add support for the multiple optima hypothesis for the thermal dependence of behavioral and physiological variables in reptiles.     

Effect of glucocorticoid hormones on fatty acid mobilization and re-esterification in rat adipose tissue

1. The effect of dexamethasone and cortisol on fatty acid mobilization and re-esterification has been studied in intact adipose tissue and isolated fat cells of the rat. 2. Dexamethasone added in vitro inhibited both the re-esterification of mobilized free fatty acids and the esterification of palmitate in the medium. 3. Under several conditions (low concentrations of dexamethasone, cortisol at a high concentration, with tissue from starved animals), steroid-induced release of free fatty acids could be accounted for by decreased re-esterification only, overall lipolytic activity remaining unmodified. At higher concentrations of dexamethasone, however, stimulation of lipolytic activity also occurred. 4. Decreased re-esterification produced by dexamethasone was observed in the total absence of glucose from the incubation medium. Further, dexamethasone stimulated the disappearance of prelabelled [(14)C]glycogen from the tissue. 5. The evidence presented suggests that mobilization of free fatty acids induced by glucocorticoid hormones under physiological conditions is primarily due to a decrease of the re-esterification rate rather than to lipase activation.     

Influence of rearing temperature on triacylglycerol storage in the pea aphid,Acyrthosiphon pisum

Total fatty acids in the pea aphid reared at low temperatures increased significantly compared to that at high rearing temperatures. This change is reflected in a large increase of myristic acid, which occurs exclusively in triacylglycerols. When aphids were moved from 25°C to a lower rearing temperature at 10°C, saturated fatty acids accumulated over time, reaching a maximum at 16th day. When aphids were moved to 4°C, a temperature below the developmental threshold, those aphids did not accumulate saturated fatty acids. Similar results were observed when aphids were exposed to sequential decrease in rearing temperature. However, both total fatty acids and myristic acid in the aphids from the treatments of sequential decreasing rearing temperature were significantly higher compared to those in the aphids from the treatments of sudden decreasing rearing temperature. This result, therefore, supports the hypothesis that cold‐adapted aphids can survive under threshold temperature for a longer period of time than noncold‐adapted aphids. Acetyl‐CoA carboxylase activity in the aphids at 25°C was twofold higher than that in the aphids at 10°C, whereas fatty acid synthase activities in the aphids reared at 25 and 10°C are similar. Aphids reared at 10°C showed a threefold reduction in reproduction rates. This reduced production of new nymphs reduces energy demand and would allow for accumulation of energy in the form of triacylglycerols. Therefore, the increased level of saturated fatty acids in aphids reared at low temperature is probably related to lower utilization of fatty acids rather than increased rates of biosynthesis.     

Heat shock‐induced metabolic conversion of membrane lipids,fatty acids and volatile organic compounds of Pyropia haitanensis under different heat shock time

Lipid metabolites play an important role in understanding the stress physiology of Pyropia haitanensis, and can be used to facilitate development of stress‐resistant Pyropia cultivars. Therefore, in this study ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC‐Q‐TOF‐MS) and gas chromatography–mass spectrometry (GC–MS) based metabolomics approaches were developed to screen the responses of lipid metabolites such as phospholipids, glycolipids, fatty acids and volatile organic compounds (VOCs) to different heat shock times. A total of 26 potential lipid biomarkers including Lyso‐monogalactosyldiacylglycerol (Lyso‐MGDG), Lyso‐digalactosyldiacylglycerol (Lyso‐DGDG), sulfoquinovosylmonoacylglycerols (SQMG), sulfoquinovosyldiacylglycerol (SQDG), diacylglyceryltrimethylhomoserine (DGTS), triacylglycerol (TAG), Lyso‐phosphatidicacid (Lyso‐PA), Lyso‐phosphatidylcholine (Lyso‐PC), Lyso‐phosphatidylethanolamine (Lyso‐PE), Lyso‐phosphatidylglycerol (Lyso‐PG), phosphatidylglycerol (PG), phosphatidylinositol (PI), and phosphatidylinositol phosphate (PIP) were identified, most of which responded to high temperature by reducing or increasing levels after stimulation for 1 h or 6 h. After times longer than 6 h, the levels of most lipids gradually recovered to the control group levels. Moreover, the balance of lipids and fatty acids transformation was disrupted. Overall, 11 total fatty acids (TFAs), 13 free fatty acids (FFAs) and 29 VOCs were identified during 0–72 h of high temperature stress. The FFAs, especially polyunsaturated C 20 fatty acids and VOCs, showed opposing change trends, indicating the transformation between C 20 fatty acids and VOCs. Overall, this study provides important insights into the metabolic variations of P. haitanensis under different heat shock time and the relationship between the conversion of lipids, fatty acids, and VOCs. The information provided herein will facilitate efficient development and improvement of Pyropia quality by producing cultivars resistant to high temperature.     

Comparison of growth, feeding and cortisol level in Platichthys bicoloratus juveniles between estuarine and nearshore nursery grounds

Growth rates estimated using the RNA : DNA ratio of field-collected stone flounder Platichthys bicoloratus juveniles were significantly higher in fish from brackish estuarine sites than from exposed nearshore sites. Gut fullness of juveniles indicated that prey availability was higher in estuaries than in nearshore areas. The whole-body cortisol concentration of juveniles, however, was approximately four times higher in fish from the estuarine than the nearshore sites. This elevated cortisol level was possibly attributable to the much higher fluctuations in temperature and salinity over a diel cycle than in nearshore habitats where both environmental factors were comparatively stable. High growth rates, feeding conditions and densities of stone flounder juveniles in estuarine nursery grounds indicated that the elevated cortisol level was within a physiologically tolerable range and did not impair fish performance.     

Comparison of rectal and tympanic core body temperature measurement in adult Guyanese squirrel monkeys (Saimiri sciureus sciureus)

Background Measuring core body temperature in a manner that is safe for animals and veterinary personnel is an important part of a physical examination. For nonhuman primates, this can involve increased restraint, additional stress, as well as the use of anesthetics and their deleterious effects on body temperature measurements. The purpose of this study was to compare two non‐invasive methods of infrared tympanic thermometry to standard rectal thermometry in adult squirrel monkeys. Methods Tympanic temperatures were collected from 37 squirrel monkeys and compared to rectal temperatures using a human and veterinary infrared tympanic thermometer. Results Compared with rectal temperature measurements, the human tympanic thermometer readings were not significantly different, while the veterinary tympanic thermometer measurements were significantly higher (P < 0.05). There were no differences between sexes. Conclusions The tympanic thermometer designed for use in humans can be used in adult squirrel monkeys as an alternative to rectal thermometry for assessing core body temperature.     

Low temperature limits photoperiod control of smolting in atlantic salmon through endocrine mechanisms

We have examined the interaction of photoperiod and temperature in regulating the parr-smolt transformation and its endocrine control. Atlantic salmon juveniles were reared at a constant temperature of 10 degrees C or ambient temperature (2 degrees C from January to April followed by seasonal increase) under simulated natural day length. At 10 degrees C, an increase in day length [16 h of light and 8 h of darkness (LD 16:8)] in February accelerated increases in gill Na(+)-K(+)-ATPase activity, whereas fish at ambient temperature did not respond to increased day length. Increases in gill Na(+)-K(+)-ATPase activity under both photoperiods occurred later at ambient temperature than at 10 degrees C. Plasma growth hormone (GH), insulin-like growth factor, and thyroxine increased within 7 days of increased day length at 10 degrees C and remained elevated for 5-9 wk; the same photoperiod treatment at 2 degrees C resulted in much smaller increases of shorter duration. Plasma cortisol increased transiently 3 and 5 wk after LD 16:8 at 10 degrees C and ambient temperature, respectively. Plasma thyroxine was consistently higher at ambient temperature than at 10 degrees C. Plasma triiodothyronine was initially higher at 10 degrees C than at ambient temperature, and there was no response to LD 16:8 under either temperature regimen. There was a strong correlation between gill Na(+)-K(+)-ATPase activity and plasma GH; correlations were weaker with other hormones. The results provide evidence that low temperature limits the physiological response to increased day length and that GH, insulin-like growth factor I, cortisol, and thyroid hormones mediate the environmental control of the parr-smolt transformation.     

Effects of isotonic and isometric exercises with mist sauna bathing on cardiovascular,thermoregulatory, and metabolic functions

To clarify the effects of isometric and isotonic exercise during mist sauna bathing on the cardiovascular function, thermoregulatory function, and metabolism, six healthy young men (22?±?1 years old, height 173?±?4 cm, weight 65.0?±?5.0 kg) were exposed to a mist sauna for 10 min at a temperature of 40 °C, and relative humidity of 100 % while performing or not performing ～30 W of isometric or isotonic exercise. The effect of the exercise was assessed by measuring tympanic temperature, heart rate, systolic and diastolic blood pressure, chest sweat rate, chest skin blood flow, and plasma catecholamine and cortisol, glucose, lactate, and free fatty acid levels. Repeated measures ANOVA showed no significant differences in blood pressure, skin blood flow, sweat rate, and total amount of sweating. Tympanic temperature increased more during isotonic exercise, and heart rate increase was more marked during isotonic exercise. The changes in lactate indicated that fatigue was not very great during isometric exercise. The glucose level indicated greater energy expenditure during isometric exercise. The free fatty acid and catecholamine levels indicated that isometric exercise did not result in very great energy expenditure and stress, respectively. The results for isotonic exercise of a decrease in lactate level and an increase in plasma free fatty acid level indicated that fatigue and energy expenditure were rather large while the perceived stress was comparatively low. We concluded that isotonic exercise may be a more desirable form of exercise during mist sauna bathing given the changes in glucose and free fatty acid levels.     

Variations of triglycerides and of plasma free fatty acids in the rat as a function of the duration of cold maintenance]

During the first day of rat exposure to cold (4 degrees C), there was a rapid and regular decrease in plasmatic triglyceride levels and an increase in the free fatty acid ones. These levels remained constant for one week then, untill the third week, they changed slowly and reached the typical levels found in cold acclimated animals. After six weeks of cold exposure, triglycerides were 2.5 fold lesser than in controls and free fatty acids were increased by 50%.     

A moderate change in temperature induces changes in fatty acid composition of storage and membrane lipids in a soil arthropod

A moderate change in ambient temperature can lead to vital physiological and biochemical adjustments in ectotherms, one of which is a change in fatty acid composition. When temperature decreases, the composition of membrane lipids (phospholipid fatty acids) is expected to become more unsaturated to be able to maintain homeoviscosity. Although different in function, storage lipids (triacylglycerol fatty acids) are expected to respond to temperature changes in a similar way. Age-specific differences, however, could influence this temperature response between different life stages. Here, we investigate if fatty acid composition of membrane and storage lipids responds similarly to temperature changes for two different life stages of Orchesella cincta. Juveniles and adults were cold acclimated (15 °C → 5 °C) for 28 days and then re-acclimated (5 °C → 15 °C) for another 28 days. We found adult membranes had a more unsaturated fatty acid composition than juveniles. Membrane lipids became more unsaturated during cold acclimation, and a reversed response occurred during warm acclimation. Membrane lipids, however, showed no warm acclimation, possibly due to the moderate temperature change. The ability to adjust storage lipid composition to moderate changes in ambient temperature may be an underestimated fitness component of temperature adaptation because fluidity of storage lipids permits accessibility of enzymes to energy reserves.     

Effect of temperature on fatty acid composition in each lipid fraction of Spirometra erinaceieuropaei plerocercoids

The effects of temperature and host fatty acids on the fatty acid contents of Spirometra erinaceieuropaei plerocercoids were investigated to clarify their role in sparganosis. After 24 hr incubation at 18 C in host snake serum, omega6 series fatty acids, especially arachidonic acid in the phospholipid fraction of the plerocercoids, increased compared with those of plerocercoids incubated at 37 C. The changes in the ratio of polyunsaturated to saturated fatty acids in the phospholipid fraction of plerocercoids incubated in physiological saline for 6 hr at 10 C were almost the same as the changes at 37 C. The ratio of polyunsaturated to saturated fatty acids of the triglyceride fraction showed almost opposite change versus the phospholipid fraction. The percentage of arachidonic acid in the phospholipid fraction of plerocercoids increased during the first 3 hr of incubation and then decreased, regardless of temperature. At 37 C, the percentage of arachidonic acid in the free fatty acid fraction fell for the first 3 hr of incubation and was significantly elevated at the end of the 6-hr incubation. At 10 C, however, arachidonic acid in the free fatty acid fraction decreased for the first hour of incubation, increased at 3 hr of incubation, then decreased again. These results suggest that fatty acids of the plerocercoids are frequently exchanged between fractions. Plerocercoids can mobilize arachidonic acid to the free fatty acid fraction more quickly at lower temperature than at higher temperature. They may utilize mobilized arachidonic acid early in the infection stage to produce prostaglandins. Alternatively, they can incorporate arachidonic acid into the phospholipid fraction again when arachidonic acid is readily available in the environment.     

Temperature effects on metabolic rate, swimming performance and condition of Pacific cod Gadus macrocephalus Tilesius

Critical swimming speed ( U _crit) and rate of oxygen consumption of Pacific cod Gadus macrocephalus acclimated to 4 and 11° C were determined to assess the influence of water temperature on performance. The physiological effect of exercise trials on fish held at two temperatures was also assessed by comparing haematocrit and plasma concentrations of cortisol, metabolites and ions collected from fish before and after testing. The U _crit of fish acclimated and exercised at 4° C did not differ from those acclimated and exercised at 11° C [1·07 body lengths (total length) s⁻¹]. While the standard metabolic rate of 11° C acclimated fish was 28% higher than that of 4° C fish, no significant difference was observed between fish acclimated at the two temperatures. Plasma concentrations of cortisol, glucose and lactate increased significantly from pre- to post-swim in both groups, yet only concentrations of cortisol differed significantly between temperature treatments. Higher concentrations of cortisol in association with greater osmoregulatory disturbance in animals acclimated at the lower temperature indicate that the lower water temperature acted as an environmental stressor. Lack of significant differences in U _crit between temperature treatments, however, suggests that Pacific cod have robust physiological resilience with respect to swimming performance within temperature changes from 4 to 11° C.     

Relationship between cardiac arrhythmia and elevated concentrations of free fatty acids in plasma after an acute myocardial ischemia in dogs]

The influence, on the heart rhythm, of an acute myocardial ischemia produced by a coronary occlusion, is studied in ten opened chest dogs. After elevation of plasmatic free fatty acids, the consequence of a second occlusion on the cardiac rhythm is analysed. There is no significant correlation between the appearance of severe ventricular arrhythmias and high plasmatic levels of free fatty acids.     

Effect of growth temperature and media composition on the fatty acid composition of Bacillus stearothermophilus AN 002

The influence of growth temperature, media composition and cell age on the chemical composition of Bacillus stearothermophilus strain AN 002 has been determined. The total cellular protein decreased and the free amino acid content increased with growth temperature, in both exponential and stationary growth phase. The protein and free amino acid contents of cells were higher in the stationary phase than in the exponential phase, irrespective of growth temperature and media composition. The RNA content was only reduced in cells grown at 55° C. No significant variations were observed in the DNA and carbohydrate contents with respect to growth temperature and cell age. The total lipid and fatty acid compositions on the other hand varied as a function of growth temperature, cell age and media composition. Differences in the relative concentrations of even, odd and branched chain fatty acids were noticed. Novariation was observed in the antiiso and unsaturated fatty acids with respect to growth temperature. The unique variations in the fatty acid composition and total lipids at the growth temperature of 50° C and their variations in the stationary growth phase seem to be characteristic for B. stearothermophilus AN 002.     

Variation of specific proteins, mitochondria and fatty acid composition in gill of Scylla serrata (Crustacea, Decapoda) under low temperature adaptation

The mud crab Scylla serrata is an important commercial crustacean inhabiting estuarine water along the coast of southeast China. Metabolism in the gill is affected continuously by fluctuating water temperature and, therefore, the ability to cope with temperature change is essential to maintain physiological function. This experiment was conducted to help understand the mechanism of low temperature adaptation in S. serrata gill. In this study, 40 healthy juvenile male S. serrata from the same broodstock were grouped randomly into four groups, which were kept at 5 °C, 10 °C, 15 °C and 27 °C, with the same feeding regime during a 3-week adaptation period. Two-dimensional electrophoresis of the proteome was conducted to separate the specific proteins responsible for low temperature adaptation. Variations in the mitochondria were observed using transmission electron microscopy, and fatty acid composition was determined using gas chromatography. The results showed that different numbers of specific proteins were expressed under different low temperature adaptation, with more expressed at 5 °C and 10 °C than at 15 °C. Mitochondrial morphology also varied under different low temperature adaptation, but there was no linear relationship between microbial density and adaptation temperature. The composition of different fatty acids in the gill varied considerably with adaptation temperature, but elongation of the carbon chain and transition from fatty acids occurred at lower temperatures. Thus, changes in the specific proteins, mitochondria and fatty acid composition of the gill were the positive effects of low temperature on metabolism, leading to improved adaptation ability in S. serrata.     

Effects of glycerol on VLDL secretion by the isolated rat liver.

Stimulation of VLDL production by increasing fatty acid availability is now well established. However, a possible regulatory role of glycerol, another lipid precursor, in VLDL synthesis by the liver has not yet been substaniated. The present experiments investigate this problem using the isolated perfused rat liver. [14C] Glycerol uptake and metabolism were studied at two different glycerol concentrations: 1 mumol/perfusate (control) or 1.6 mmol/perfusate. VLDL production and lipid synthesis were investigated using [14C]leucine and several labelled fatty acids as precursors in control and glycerol-overloaded livers. Neoglycogenesis and lipogenesis from glycerol carbons are negligible in our conditions. The absolute amount of glycerol, but not the precentage, taken up by the liver, increased after raising its concentration in the perfusate. A major part of exogenous (plasmatic) glycerol was esterified with endogenous (non plasmatic) fatty acids. Incorporation of radioactive fatty acids into liver or plasma lipids was lower than in the the control group. Significant differences were observed between saturated and unsaturated fatty acids used as lipid precursors. Production of VLDL as assessed by radioactive leucine and fatty acid incorporation in the VLDL of the perfusate was depressed by glycerol. Glycerol partly inhibits the normal stimulation of VLDL production by plasmatic fatty acid overload.     

Short-term physiological responses to moderate heat stress in grazing dairy cows in temperate climate

Even in temperate climate regions, an increase in ambient temperature and exposure to solar radiation can cause heat stress in lactating dairy cows. We hypothesised that grazing dairy cows exhibit short-term physiological changes due to increasing heat load under moderate climate conditions. Over two consecutive summers, 38 lactating Holstein dairy cows were studied in a full-time grazing system. Data were collected in 10 experimental periods of up to three consecutive days with a moderate comprehensive climate index (CCI). The individual animals’ vaginal temperature (VT), heart rate, and locomotor activity data were automatically monitored with sensors. Blood samples and proportional whole milk samples were collected at afternoon milking. The concentrations of beta-hydroxybutyrate, glucose, non-esterified fatty acids, urea nitrogen, plasma thyroxine and triiodothyronine were analysed in blood plasma, and fat, protein, lactose, urea nitrogen, cortisol, Na⁺, K⁺, and Cl^? concentrations were analysed in milk. The daily distribution of VT recordings greater than 39 °C showed a circadian rhythm with a proportion of recordings of 2% and lower during the night and a percentage of 10% or higher in the afternoon. The cows’ maximal daily vaginal temperature (VT_MAX) between 0830 and 1430 h was positively related to the mean daily CCI in the same time period (CCI_MEAN; mean and SD 23.6 ± 5.4 °C). Cows with greater VT_MAX had an increased mean heart rate, plasma glucose and milk cortisol concentrations and decreased concentrations of plasma thyroxine and triiodothyronine. The concentration of Na⁺ in milk was lower, and the concentration of K⁺ in milk tended to be higher in cows with increased VT_MAX. For beta-hydroxybutyrate, non-esterified fatty acids and urea nitrogen concentrations in plasma and fat and lactose concentrations in milk no relationships were found in terms of increasing VT. For milk urea nitrogen and protein concentrations, the proportion of total variance explained by inter-individual or -period variance was high. In conclusion, changes observed in milk and blood likely reflected short-term physiological responses to moderate heat stress. In particular, milk cortisol and Na⁺ may be useful traits for timely monitoring of heat stress in individual cows because their inter-individual variances were relatively small and samples can be collected non-invasively.     

Determination of plasma free fatty acids, free cholesterol, cholesteryl esters, and triacylglycerols directly from total lipid extract by capillary gas chromatography

An accurate capillary gas chromatographic method using different internal standards for determining free fatty acids, cholesterol, cholesteryl esters, and triacylglycerols in plasma and other biological sources is described. It is designed to give information about species composition and, consequently, more detailed information about changes in lipid metabolism of patients suffering from metabolic disorders. After plasma extraction the lipids, except phospholipids, are directly examined without any further derivatization. For free fatty acid determination the programmed temperature vaporizer (PTV) injector was heated from 40 degrees C (sample introduction) to 190 degrees C. In a second gas chromatographic run the PTV-injector system was heated from 60 degrees C (sample introduction) to 400 degrees C, enabling the determination of free cholesterol, cholesteryl esters, and triacylglycerol species, differing in the number of carbon atoms. Evaluation of the values obtained resulted in coefficients of variation (%) of 1.0-2.8, 2.0, 1.29-2.24, and 2.8, for free fatty acid standards, plasma free fatty acids, cholesterol and cholesteryl ester standards, and plasma total cholesterol, respectively. Free fatty acids, cholesterol, and cholesteryl esters were not influenced by storage of plasma at -24 degrees C up to 4 days prior to extraction. The results of the gas chromatographic method and the enzymatic methods correlated well. Determination by gas chromatography yielded higher total cholesterol and lower triacylglycerol values than those values obtained by enzymatic methods.     

Haematology and serum biochemistry of captive unsedated chital deer (Axis axis) in Australia

Haematological and serum biochemical values were measured in blood samples collected over a 12-mo period from 37 unsedated chital deer (Axis axis). Stags and hinds, ranging in age from birth to maturity, were sampled. Haemoglobin, total erythrocyte and haematocrit values were low at birth and sex differences were not apparent in neonates and juveniles, but were in adults. Chital stags had higher erythrocyte parameters (P less than 0.001) and lower erythrocyte indices than hinds, and the total leucocyte count was higher in stags (P less than 0.01). Some parameters (erythrocytes, muscle enzymes, glucose, cortisol) decreased over successive serial sampling. The differential leucocyte count of older stags decreased during the initial handling period. The major rutting period in February and March was characterised by changes in the differential leucocyte count, elevations in serum muscle enzymes, and lower serum cortisol levels. Alkaline phosphatase activity in serum reflected the annual antler cycle of chital stags. Serial sampling over many weeks, either weekly or tri-weekly, produced haematological and biochemical changes in successive samples which may have reflected a reduction in stress and excitement associated with restraint.