Shivering modulation in humans: Effects of rapid changes in environmental temperature

During cold exposure, increase in heat production is produced via the activation of shivering thermogenesis and nonshivering thermogenesis, the former being the main contributor to compensatory heat production in non-acclimatized humans. In rats, it has been demonstrated that shivering thermogenesis is modulated solely by skin thermoreceptors but this modulation has yet to be investigated in humans. The aim of this study was to determine if cold-induced shivering in humans can be modulated by cutaneous thermoreceptors in conditions where increases in heat loss can be adequately compensated by increases in thermogenic rate. Using a liquid-conditioned suit, six non-acclimatized men were exposed to cold (6 °C) for four 30 min periods, each of them separated by 15 min of heat exposure (33 °C). Core temperature remained stable throughout exposures whereas skin temperatures significantly decreased by 12% in average during the sequential cold/heat exposures compared to baseline (p<0.0001). Shivering intensity and metabolic rate increased significantly during 6 °C exposures (3.3±0.7% MVC, 0.40±0.0 L O₂/min, respectively) and were significantly reduced during 33 °C exposure (0.5±0.1% MVC, 0.25±0.0 L O₂/min; p<0.005 for both). Most importantly, shivering could be quickly and strongly inhibited during 33 °C exposure although skin temperature often remained below baseline values. In conclusion, under compensatory conditions, cutaneous thermoreceptors appear to be a major modulator of the shivering response in humans and seem to react rapidly to changes in the microclimate right next to the skin and to skin temperature.     

Feedback controlled force enhancement and activation reduction of voluntarily activated quadriceps femoris during sub-maximal muscle action

Stretch of activated muscles leads to enhanced forces compared to isometric contractions at the same muscle length and the same level of activation. This so-called residual force enhancement (RFE) is thought to be a property of all muscles and preparations. However, observations concerning the existence, amount and duration of RFE are inconsistent, especially for voluntary activated large human muscles. Therefore, physiological relevance for daily activity is still questionable and the purpose of this study was to examine whether RFE is present in voluntary sub-maximal activated quadriceps femoris (QF). Seated in a rotational dynamometer with EMG attached to superficial parts of QF, 30 subjects performed isometric and isometric-eccentric-isometric contractions (20° stretch, ω = 60° s⁻¹) at 30% and 60% of maximum voluntary activation (MVA) and contraction (MVC). To account for the complexity of the multi-headed QF, a compensation model based on physiological cross-sectional area and individual EMG-torque relations was used to interpret EMG data. For both levels of intensity and both feedback control strategies, ANOVA identified significant RFE (at the same level of activation) and reduced activation (at the same level of torque). Against expectations, RFE was independent of the level of activation.     

Effect of innervation zones in estimating biceps brachii force-EMG relationship during isometric contraction

Measuring muscle forces in vivo is invasive and consequently indirect methods e.g., electromyography (EMG) are used in estimating muscular force production. The aim of the present paper was to examine what kind of effect the disruption of the physiological signal caused by the innervation zone has in predicting the force/torque output from surface EMG. Twelve men (age 26 (SD ±3) years; height 179 (±6) cm; body mass 73 (±6) kg) volunteered as subjects. They were asked to perform maximal voluntary isometric contraction (MVC) in elbow flexion, and submaximal contractions at 10%, 20%, 30%, 40%, 50% and 75% of the recorded MVC. EMG was measured from biceps brachii muscle with an electrode grid of 5 columns × 13 rows. Force-EMG relationships were determined from individual channels and as the global mean value. The relationship was deemed inconsistent if EMG value did not increase in successive force levels. Root mean squared errors were calculated for 3rd order polynomial fits. All subjects had at least one (4-52) inconsistent channel. Two subjects had inconsistent relationship calculated from the global mean. The mean root mean squared error calculated using leave one out method for the fits of the individual channels (0.33 ± 0.17) was higher (P < 0.001) than the error for the global mean fit (0.16 ± 0.08). It seems that the disruption of the physiological signal caused by the innervation zone affects the consistency of the force-EMG relationship on single bipolar channel level. Multichannel EMG recordings used for predicting force overcame this disruption.     

Effects of local elastic compression on muscle strength, electromyographic, and mechanomyographic responses in the lower extremity

The purpose of this study was to investigate the effect of elastic compression on muscle strength, electromyographic (EMG), and mechanomyographic (MMG) responses of quadriceps femoris during isometric and isokinetic contractions. Twelve participants performed 5 s isometric maximal voluntary contractions (MVC) and 25 consecutive and maximal isokinetic knee extensions at 60 and 300 °/s with no (control, CC), medium (MC), and high (HC) compression applied to the muscle. The EMG and MMG signals were collected simultaneously with muscle isometric and isokinetic strength data. The results showed that the elevated compression did not improve peak torque, peak power, average power, total work, and regression of torque in the isometric and isokinetic contractions. However, the root mean squared value of EMG in both HC and MC significantly decreased compared with CC at 60 and 300 °/s (p < 0.01). Furthermore, the EMG mean power frequency in HC was significantly higher than that in CC at 60 °/s (p < 0.05) whereas no significant compression effect was found in the MMG mean power frequency. These findings provide preliminary evidence suggesting that the increase in local compression pressure may effectively increase muscle efficiency and this might be beneficial in reducing muscle fatigue during concentric isokinetic muscle contractions.     

The effect of thermopreference on circadian thermoregulation in sprague-dawley and fisher 344 rats

Interstrain differences in thermoregulation of rats are important in biomedical research because subtleties in thermoregulatory sensitivities may greatly affect data collected. Little is known regarding how individual rodent strains differentially utilize behavioral thermal preference to regulate core temperature (Tc). Sprague-Dawley (SD) and Fischer 344 (F344) rats are known to have differences in thermoregulation including heat tolerance and are useful models to study interstrain differences in thermoregulation. Adult male SD and F344 rats of similar body size were implanted with radiotelemetry thermoprobes (DSI) to measure Tc and MA and housed in either a longitudinal temperature gradient with an ambient temperature (Ta) range of ∼15–40 °C to measure selected Ta (STa) or control environment maintained at a Ta of 23 °C. When continuously monitored for 48 h, Tc and MA increased at night, while STa decreased, according to their normal circadian cycle in both strains. SD rats were more active than F344 rats throughout the circadian cycle (SD gradient: day=12.9±1.2 m/h, night=32.1±2.4 m/h; F344 gradient: day=4.1±0.6 m/h, night=16.8±1.8 m/h; p<0.05 interstrain and circadian effects). The STa of each strain was greater during the daytime (SD: 26.4±0.2 °C; F344: 27.8±0.3 °C) than at night (SD: 24.7±0.3 °C; F344: 25.7±0.3 °C) confirming past studies that thermopreference during the day and night is greater than standard room temperature (∼23 °C). Correlations between MA and Tc suggest that MA has a greater effect on Tc in the F344 but not the SD strain when housed in a temperature gradient. There were significant strain differences in Tc depending on whether rats were housed in a temperature gradient. That is, the control F344 rats had a lower Tc during the transition from dark to light compared to rats housed in a gradient. Tc of the SD strain was unaffected by housing in the gradient. Rats are typically housed at a standard room temperature of 23 °C. However, the results demonstrate that when given the opportunity to behaviorally thermoregulate in a temperature gradient, the F344 strain selects a warmer environment that affects the regulation of Tc. This may be important in the experimenters' choice of ambient temperatures to house and study rats and other rodents.     

Suppression of MAPKAPK2 during mammalian hibernation

Metabolic signaling coordinates the transition by hibernating mammals from euthermia into profound torpor. Organ-specific responses by activated p38 mitogen activated protein kinase (MAPK) are known to contribute to this transition. Therefore, we hypothesized that the MAPK-activated protein kinase-2 (MAPKAPK2), a downstream target of p38 MAPK, would also be active in establishing the torpid state. Kinetic parameters of MAPKAPK2 from skeletal muscle of Richardson’s ground squirrels, Spermophilus richardsonii, were analyzed using a fluorescence assay. MAPKAPK2 activity was 27.4 ± 1.27 pmol/min/mg in muscle from euthermic squirrels and decreased by ∼63% during cold torpor, while total protein levels were unchanged (as assessed by immunoblotting). In vitro treatment of MAPKAPK2 via stimulation of endogenous phosphatases and addition of commercial alkaline phosphatase decreased enzyme activity to only ∼3–5% of its original value in muscle extracts from both euthermic and hibernating squirrels suggesting that posttranslational modification suppresses MAPKAPK2 during the transition from euthermic to torpid states. Enzyme S_0.5 and n_H values for ATP and peptide substrates changed significantly between euthermia and torpor, and also between assays at 22 versus 10 °C but, kinetic parameters were actually closely conserved when values for the euthermic enzyme at 22 °C were directly compared with the hibernator enzyme at 10 °C. Arrhenius plots showed significantly different activation energies of 40.8 ± 0.7 and 54.3 ± 2.7 kJ/mol for the muscle enzyme from euthermic versus torpid animals, respectively but MAPKAPK2 from the two physiological states showed no difference in sensitivity to urea denaturation. Overall, the results show that total activity of MAPKAPK2 is in fact reduced, despite previous findings of p38 MAPK activation, and kinetic parameters are altered when ground squirrels enter torpor but protein stability is not apparently changed. The data suggest that MAPKAPK2 suppression may have a significant role in the differential regulation of muscle target proteins when ground squirrels enter torpor.     

Heat tolerance, behavioural temperature selection and temperature-dependent respiration in larval Octopus huttoni

This study reports temperature effects on paralarvae from a benthic octopus species, Octopus huttoni, found throughout New Zealand and temperate Australia. We quantified the thermal tolerance, thermal preference and temperature-dependent respiration rates in 1-5 days old paralarvae. Thermal stress (1 °C increase h⁻¹) and thermal selection (∼10-24 °C vertical gradient) experiments were conducted with paralarvae reared for 4 days at 16 °C. In addition, measurement of oxygen consumption at 10, 15, 20 and 25 °C was made for paralarvae aged 1, 4 and 5 days using microrespirometry. Onset of spasms, rigour (CT_max) and mortality (upper lethal limit) occurred for 50% of experimental animals at, respectively, 26.0±0.2 °C, 27.8±0.2 °C and 31.4±0.1 °C. The upper, 23.1±0.2 °C, and lower, 15.0±1.7 °C, temperatures actively avoided by paralarvae correspond with the temperature range over which normal behaviours were observed in the thermal stress experiments. Over the temperature range of 10 °C-25 °C, respiration rates, standardized for an individual larva, increased with age, from 54.0 to 165.2 nmol larvae⁻¹ h⁻¹ in one-day old larvae to 40.1-99.4 nmol h⁻¹ at five days. Older larvae showed a lesser response to increased temperature: the effect of increasing temperature from 20 to 25 °C (Q₁₀) on 5 days old larvae (Q₁₀=1.35) was lower when compared with the 1 day old larvae (Q₁₀=1.68). The lower Q₁₀ in older larvae may reflect age-related changes in metabolic processes or a greater scope of older larvae to respond to thermal stress such as by reducing activity. Collectively, our data indicate that temperatures >25 °C may be a critical temperature. Further studies on the population-level variation in thermal tolerance in this species are warranted to predict how continued increases in ocean temperature will limit O. huttoni at early larval stages across the range of this species.     

High-volume intermittent maximal intensity isometric exercise caused great stress,although central motor fatigue did not occur

To establish whether very high-volume, high-intensity isometric exercise causes stress to the body and how it affects peripheral and central fatigue. Nineteen physically active healthy male subjects (21.2 ± 1.7 years; height – 1.82 ± 0.41 m, body weight – 79.9 ± 4.5 kg; body mass index – 24.3 ± 2.1 kg/m2) volunteered to participate in this study. They participated in two experiments 3–5 days apart. Each experiment comprised six series of 60-s maximum voluntary contraction (MVC) force (knee extension) achieved as rapidly as possible. This very high-volume, high-intensity exercise (HVHIE) was performed at different quadriceps muscle lengths: short (SL) and long (LL). The MVC and the electrically stimulated contractile properties of the muscle were measured prior to HVHIE, immediately after and 3 min after each series, and at 3, 10, and 30 min after the end of HVHIE. We found that HVHIE caused high levels of stress (cortisol levels approximately doubled, heart rate and the root mean square successive difference of interval (RMSSD) decreased by about 75%); lactate increased to 8–11 mmol/L, voluntary and 100 Hz stimulation-induced force (recorded immediately after HVHIE) decreased by 55% at LL and 40% at SL. However, the central activation ratio during MVC did not change after either exercise. Isometric HVHIE performed using one leg caused high levels of stress (RMSSD decreased, cortisol increased after HVHIE equally at SL and LL; La increased more while exercising at LL) and the voluntary and electrostimulation-induced muscle force significantly decreased, but muscle central activation during MVC did not decrease.     

Can programmed cell death be induced in post-ejaculatory bull and stallion spermatozoa?

Apoptosis is common during spermatogenesis. Here, it was tested whether apoptosis could be induced in sperm after ejaculation. There were several lines of evidence to indicate that sperm are resistant to induction of apoptosis. First, incubation of bull sperm at temperatures characteristic of normothermia (38.5 °C) or heat shock (40 and 41 °C) for 4 h did not increase the proportion of sperm positive for the TUNEL reaction. There was also no reduction in mitochondrial polarity caused by exposure to 40 or 41 °C. Incubation at 38.5 °C (least-squares mean ± SEM = 4.0 ± 1.4%), 40 °C (6.2 ± 1.4%), and 41 °C (7.0 ± 1.4%) for 24 h did increase the proportion of sperm that were TUNEL positive slightly as compared to non-incubated control sperm (1.0 ± 1.4%). However, the increase in TUNEL labeling was not affected by incubation temperature and occurred even in the presence of the group II caspase inhibitor, z-DEVD-fmk. In addition, exposure of bull sperm to carbonyl cyanide 3-chlorophenylhydrazone (CCCP), which depolarizes mitochondrial membranes, did not increase TUNEL labeling. Stallion sperm were also resistant to increased TUNEL labeling in response to incubation at 41 °C for 4 h or exposure to CCCP. Western blotting was performed to determine whether failure of induction of apoptosis was due to aberrant caspase activation. Procaspase-9 was detected in bull sperm, but cleavage to caspase-9 was not induced by short-term aging at 38.5, 40, or 41 °C, or exposure to CCCP. Procaspase-3 was not detected in bull spermatozoa. In conclusion, post-ejaculatory bull and stallion sperm were resistant to induction of apoptosis; this resistance, at least in bulls, was due to refractoriness of mitochondria to heat shock-induced depolarization, lack of activation of procaspase-9, and an absence of procaspase-3.     

Effect of temperature on isoproterenol-induced increases in left ventricular developed pressure

The goal of this project was to determine the effects of elevated cardiac temperature on preload-dependent and preload-independent regulation of left ventricular developed pressure (LVDP) in Langendorff-perfused, electrically paced (420 bpm), Sprague-Dawley rat hearts. LVDP responses to steady-state isoproterenol infusions (10⁻⁸ M) were determined at 37, 38, 39, and 40 °C. Preload-dependent LVDP was determined at 37 and 40 °C. Isoproterenol-induced LVDP and preload-dependent LVDP time controls were conducted in a separate group maintained at 37 °C. The percent increase in LVDP during isoproterenol infusion significantly decreased at 40 °C to 42±6 (SE), compared to 55±9, 55±6, and 53±7% at 37, 38, and 39 °C, respectively. No significant differences were observed in the percent increase in LVDP to isoproterenol among the corresponding time controls (50±6, 47±3, 56±4, and 56±5%). Preload-dependent LVDP decreased across the experimental protocol, but there were no cardiac temperature effects. These data indicate that β-adrenergic mediated contractility is not altered by moderate heating from normothermia but is compromised at very high temperatures (40 °C). Cardiac temperatures from 37 to 40 °C do not alter the inherent preload-dependent LVDP, indicating that the Frank–Starling relation is not directly affected within this temperature range.     

Effects of heat acclimatisation on work tolerance and thermoregulation in trained tropical natives

This study aimed to investigate the effects of heat acclimatisation on thermoregulatory responses and work tolerance in trained individuals residing in the tropics. Eighteen male trained soldiers, who are native to a warm and humid climate, performed a total of four heat stress tests donning the Skeletal Battle Order (SBO, 20.5 kg) and Full Battle Order (FBO, 24.7 kg) before (PRE) and after (POST) a 10-day heat acclimatisation programme. The trials were conducted in an environmental chamber (dry bulb temperature: 32 °C, relative humidity: 70%, solar radiation: 400 W/m²). Excluding the data sets of which participants fully completed the heat stress tests (210 min) before and after heat acclimatisation, work tolerance was improved from 173±30 to 201±18 min (∼21%, p<0.05, n=9) following heat acclimatisation. Following heat acclimatisation, chest skin temperature during exercise was lowered in SBO (PRE=36.7±0.3 vs. POST=36.5±0.3 °C, p<0.01) and FBO (PRE=36.8±0.4 vs. POST=36.6±0.3 °C, p<0.01). Ratings of perceived exertion were decreased with SBO and FBO (PRE=11±2; POST=10±2; p<0.05) after heat acclimatisation. Heat acclimatisation had no effects on baseline body core temperature, heart rate and sweat rate across trials (p>0.05). A heat acclimatisation programme improves work tolerance with minimal effects on thermoregulation in trained tropical natives.     

Effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration,ROS production and lipid peroxidation in broiler chickens

In order to investigate the effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration, production of reactive oxygen species (ROS) and lipid peroxidation in broiler chickens, 128 six-week-old broiler chickens were kept in a controlled-environment chamber. The broiler chickens were initially kept at 25 °C (relative humidity, RH, 70 ± 5%) for 6 d and subsequently exposed to 35 °C (RH, 70 ± 5%) for 3 h, then the heat stress was removed and the temperature returned to 25 °C (RH, 70 ± 5%). Blood and liver samples were obtained before heat exposure and at 0 (at the end of the three-hour heating episode, this group is also abbreviated as the HT group), 1, 2, 4, 8, 12 h after the stress was removed. The results showed that acute heat stress induced a significant production of ROS, function of the mitochondrial respiratory chain, antioxidative enzymes [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px)] activity, and formation of malondialdehybe (MDA). Within the first 12 h after removal of the heat stress, the acute modification of the above parameters induced by heat stress gradually approached to pre-heat levels. The results of the present study suggest that acute exposure to high temperatures may depress the activity of the mitochondrial respiratory chain. This leads to over-production of ROS, which ultimately results in lipid peroxidation and oxidative stress. When the high temperature was removed, the production of ROS, mitochondrial respiratory function and oxidative injury that were induced by acute heat exposure gradually approached the levels observed before heating, in a time-dependent manner.     

Effect of cold on lipid peroxidation in the brown adipose tissue and liver of rats

1. Lipid peroxidation in the interscapular brown adipose tissue (iBAT) and liver was studied in rats acclimated to room (23±1 °C) and low temperature (5±1 °C, 42 days), as well as in animals exposed to 5±1 °C for 24 h; in addition, the tissue metallothionein (MT) and iron were determined.     

Quadriceps neuromuscular function and self-reported functional ability in knee osteoarthritis

The purposes of this study were to determine 1) the relationships of self-reported function scores in patients with knee osteoarthritis (OA) to both maximal isometric torque and to isotonic power at a variety of loads, and 2) the degree to which muscle volume (MV) or voluntary activation (VA) are associated with torque and power measures in this population. Isometric maximal voluntary contraction (MVC) torque and isotonic power [performed at loads corresponding to 10, 20, 30, 40, and 50% MVC, and a minimal load ("Zero Load")] were measured in 40 participants with knee OA. Functional ability was measured with the Western Ontario and McMaster Osteoarthritis Index (WOMAC) function subscale. MV was determined with magnetic resonance imaging, and VA was measured with the interpolated twitch technique. In general, power measured at lower loads (Zero Load and 10-30% MVC, r(2) = 0.21-0.28, P < 0.05) predicted a greater proportion of the variance in function than MVC torque (r(2) = 0.18, P < 0.05), with power measured at Zero Load showing the strongest association (r(2) = 0. 28, P < 0.05). MV was the strongest predictor of MVC torque and power measures in multiple regression models (r(2) = 0.42-0.72). VA explained only 6% of the variance in MVC torque and was not significantly associated with power at any load (P > 0.05). Quadriceps MVC torque and power are associated with self-reported function in knee OA, but muscle power at lower loads is more predictive of function than MVC torque. The variance in MVC torque and power between participants is due predominantly to differences in MV and has little to do with deficits in VA.     

Postactivation potentiation in a human muscle: effect on the rate of torque development of tetanic and voluntary isometric contractions.

Postactivation potentiation (PAP), a mechanism by which the torque of a muscle twitch is increased following a conditioning contraction, is well documented in muscular physiology, but little is known about its effect on the maximal rate of torque development and functional significance during voluntary movements. The objective of this study was to investigate the PAP effect on the rate of isometric torque development of electrically induced and voluntary contractions. To that purpose, the electromechanical responses of the thumb adductor muscles to a single electrical stimulus (twitch), a train of 15 pulses at 250 Hz (HFT(250)), and during ballistic (i.e., rapid torque development) voluntary contractions at torque levels ranging from 10 to 75% of maximal voluntary contraction (MVC) were recorded before and after a conditioning 6-s MVC. The results showed that the rate of torque development was significantly (P < 0.001) increased after the conditioning MVC, but the effect was greater for the twitch ( approximately 200%) compared with the HFT(250) ( approximately 17%) or ballistic contractions (range: 9-24%). Although twitch potentiation was maximal immediately after the conditioning MVC, maximal potentiation for HFT(250) and ballistic contractions was delayed to 1 min after the 6-s MVC. Furthermore, the similar degree of potentiation for the rate of isometric torque development between tetanic and voluntary ballistic contractions indicates that PAP is not related to the modality of muscle activation. These observations suggest that PAP may be considered as a mechanism that can influence our contractions during daily tasks and can be utilized to improve muscle performance in explosive sports.     

Duration tenacity: A method for assessing acclimatory capacity of the Antarctic limpet, Nacella concinna

The limpet, Nacella concinna, collected from the Antarctic Peninsula (67°S), was incubated at − 0.3 °C and 2.9 °C for 9 months to test if the previously reported absence of acclimation capacity in Antarctic marine ectotherms could be due to the extended time it takes for them to adjust their physiology to a new stable state. Acclimation was tested through acute measurements of upper lethal limit and a modified measure of tenacity, that tested muscle capacity by measuring the length of time that N. concinna were able to remain attached to the substratum at different temperatures. Both measures acclimated in response to incubation to the higher temperature. Lethal limits were elevated in N. concinna incubated at 2.9 °C (8.1 ± 0.3 °C) compared to those incubated at − 0.3 °C (6.9 ± 0.4 °C). 2.9 °C incubated N. concinna also had a maximum tenacity at 2.1 °C, a higher temperature than the maximum tenacity of those incubated at − 0.3 °C, which occurred at − 1.0 °C. This study is the first to show that the Antarctic limpet can acclimate its physiology, but that it requires a greater period of time for acclimation to occur than previous studies have allowed for.     

Spatial EMG potential distribution pattern of vastus lateralis muscle during isometric knee extension in young and elderly men

The aim of the present study was to compare spatial electromyographic (EMG) potential distribution during force production between elderly and young individuals using multi-channel surface EMG (SEMG). Thirteen elderly (72-79 years) and 13 young (21-27 years) healthy male volunteers performed ramp submaximal contraction during isometric knee extension from 0% to 65% of maximal voluntary contraction. During contraction, multi-channel EMG was recorded from the vastus lateralis muscle. To evaluate alteration in heterogeneity and pattern in spatial EMG potential distribution, coefficient of variation (CoV), modified entropy and correlation coefficients with initial torque level were calculated from multi-channel SEMG at 5% force increment. Increase in CoV and decrease in modified entropy of RMS with increase of exerted torque were significantly smaller in elderly group (p < 0.05) and correlation coefficients with initial torque level were significantly higher in elderly group than in young group at moderate torque levels (p < 0.05). These data suggest that the increase of heterogeneity and the change in the activation pattern are smaller in elderly individuals than in young individuals. We speculated that multi-channel SEMG pattern in elderly individual reflects neuromuscular activation strategy regulated predominantly by clustering of similar type of muscle fibers in aged muscle.     

Remote sensing of physiological data: Impact of long term captivity on body temperature variation of the feral cat (Felis catus) in Australia,recorded via Thermochron iButtons

This study reports body temperature regulation (T_b) and circadian rhythms of undisturbed feral cats in their natural environment in Australia over a continuous period of three months. It furthermore compares these data with T_b data collected of feral cats, after a period of one year in captivity. In free-ranging, undisturbed feral cats, a distinct robust, regular circadian rhythm (strength of rhythm) (21–59.8%) with higher body temperature in the dark (active) phase (mean±STD: 39.2±0.27 °C) and significantly lower body temperature during the light (rest) phase (mean±STD: 38.1±0.47 °C, P<0.001) was found. The acrophase (time of the daily peak) of the three free-ranging cats investigated varied from 22:34 h (LG 2), 22:57 h (LG 1) to 23:17 h (LG 3). In the course of captivity, the cats’ circadian rhythms shifted from nocturnality to a diurnal tendency, with an acrophase ranging from 12:00 h (MtK 2), 12:23 h (MtK 1) to 16:25 h (MtK 3). This change in rhythmicity was accompanied by a significant decrease in robustness (1.7–5.2%) and mean body temperature levels (37.77±0.34 °C) as well as minima and maxima (36–39 °C versus 35.5–41.9 °C, free-ranging cats) of three captive cats, resulting in a significant shift towards a decrease in amplitude.     

Evaporative water loss and energy metabolic in two small mammals,voles (Eothenomys miletus) and mice (Apodemus chevrieri), in Hengduan mountains region

Evaporative water loss (EWL) and energy metabolism were measured at different temperatures in Eothenomys miletus and Apodemus chevrieri in dry air. The thermal neutral zone (TNZ) of E. miletus was 22.5–30 °C and that of A. chevrieri was 20–27.5 °C. Mean body temperatures of the two species were 35.75±0.5 and 36.54±0.61 °C. Basal metabolic rates (BMR) were 1.92±0.17 and 2.7±0.5 ml O₂/g h, respectively. Average minimum thermal conductance (C_m) were 0.23±0.08 and 0.25±0.06 ml O₂/g h °C. EWL in E. miletus and A. chevrieri increased with the increase in temperature; the maximal EWL at 35 °C was 4.78±0.6 mg H₂O/g h in E. miletus, and 5.92±0.43 mg H₂O/g h in A. chevrieri. Percentage of evaporative heat loss to total heat production (EHL/HP) increased with the increase in temperature; the maximal EHL/HP was 22.45% at 30 °C in E. miletus, and in A. chevrieri it was 19.96% at 27.5 °C. The results may reflect features of small rodents in the Hengduan mountains region: both E. miletus and A. chevrieri have high levels of BMR and high levels of total thermal conductance, compared with the predicted values based on their body masses, while their body temperatures are relatively low. EWL plays an important role in temperature regulation.     

Cryopreservation of red snapper (Lutjanus argentimaculatus) sperm: Effect of cryoprotectants and cooling rates on sperm motility, sperm viability, and fertilization capacity

Sperm cryopreservation of red snapper (Lutjanus argentimaculatus) is essentially unexplored, although many species of the Lutjanidae family are considered to be high-value commercial species. The objective of this study was to develop a species-specific cryopreservation protocol for red snapper (L. argentimaculatus) sperm by optimizing cryoprotectants and cooling rates in the cryopreservation procedure. Ten cryoprotectants at four concentrations and two freezing protocols were examined in two separate experiments. In the first experiment, toxicity studies of dimethyl sulfoxide (DMSO), glycerol, propylene glycol (PG), ethylene glycol (EG), formamide, methanol, ethanol, sucrose, trehalose, and dimethylacetamide (DMA) on sperm motility were performed. Semen diluted 1:1 in Ringer solution were exposed to cryoprotectants at four final concentrations of 5%, 10%, 15%, or 20% for periods of 10, 20, 30, 40, 50, 60, 90, and 120 min at room temperature (25 °C). The cryoprotectants and concentrations that showed the least toxic effect on sperm motility were selected for cryopreservation trials. In the second experiment, selected cryoprotectants were then assessed for freezing capacity of sperm as follows: DMSO 5% and 10%, PG 5% and 10%, EG 5% and 10%, ethanol 5%, and methanol 5%. Semen was diluted 1:1 in Ringer solution and equilibrated with selected cryoprotectants for 10 min at room temperature. Sperm were frozen in a controlled-rate programmable freezer at four cooling rates of 3, 5, 10, and 12 °C/min from an initial temperature of 25 °C to final temperatures of −40 or −80 °C before plunging into liquid nitrogen. Sperm equilibrated in 10% DMSO and cooled at a rate of 10 °C/min to a final temperature of −80 °C had the highest motility (91.1 ± 2.2%) and viability (92.7 ± 2.3%) after thawing. The fertilization rate of frozen-thawed sperm (72.4 ± 2.4%) was not different (P > 0.05) from that of fresh sperm (75.5 ± 2.4%). This study apparently represents the first reported attempt for cryopreservation of L. argentimaculatus sperm.