Do newborn lambs with black and white hair-coats in an equatorial semi-arid environment maintain homeothermy?

We assessed newborn lambs from two hair-coat sheep breeds, the black Santa Ines (n = 29) and white Dorper (n = 26), to determine how they behaviourally and physiologically respond to the prevailing thermal conditions in an equatorial semi-arid environment. Measurements of hair-coat surface temperature, rectal temperature and the lambs' exposure to sun were recorded across the first 24 h of life every hour, after the lambs had received colostrum. Lambs and ewes were kept in a lambing pen and could freely move between a shaded area or be exposed to sun. During the study period, the air temperature ranged between 20 ºC and 34 ºC. When exposed to sun, lambs and ewes could experience as much as 1200 W m-2 of solar irradiance. Santa Ines lambs exhibited higher (highest density interval at 95%, HDI) hair-coat surface temperatures than did Dorper lambs. Overall, both Santa Ines and Dorper lambs sustained homeothermy, with a mean rectal temperature ranging from 38.7 ºC to 39.1 ºC between night and daytime phase, and a mean amplitude of ~ 0.8 ºC. Nevertheless, from 10:00 to 15:00, some lambs were found to be moderately hyperthermic. Five Santa Ines and three Dorper lambs had rectal temperatures above 40 ºC, and one Santa Ines lamb, while exposed to sun, had a rectal temperature of 41.3 ºC. Over this time period, lambs were more likely to move to shade (HDI at 95%). From 00:00 to 06:00, despite the air temperature being lower than the hair-coat surface, favouring high rates of sensible heat loss to the environment, no lamb exhibited signs of hypothermia (rectal temperature < 37.5 ºC). In conclusion, haired newborn lambs coped well with high levels of radiant heat during the daytime and lower temperatures at night. However, providing access to shade during the daytime is important to improve the welfare of newborn lambs delivered in equatorial semi-arid regions.

Energy cost of physical activities in growing broilers.

1. The time-energy budget method estimates the energy used for physical activity (PA) by integrating behaviour PA patterns with energy cost for specific PAs. Nevertheless, information about individual energy cost by type of PA are not available and so this study estimated the energy cost of PA for growing broilers.2. An indirect calorimetry system for single birds was constructed to measure the variation in the rate of O2 consumption (V˙O2, L/min) and rate of CO2 production (V˙CO2, L/min) produced by these PAs.3. A total of five birds were used in a replicated trial where their body weight (BW) ranged from 1.5 to 2.5 kg to measure the increase in heat production (HP) above resting levels as a result of PA. The procedure in the chamber was divided into five steps: (1) initial baselining, (2) resting metabolic rate, (3) PA such as feeding, drinking and other standing activities, (4) removal of gas exchange produced in step 3, and (5) final baselining. The PA was recorded using a video camera fixed at the chamber's top (and outside).4. The area under V˙CO2 and V˙O2 curves was used to calculate the CO2 production (vCO2, L) and O2 consumption (vO2, L). Then, the HP (cal/kg-0.75) was calculated according to the Brouwer equation. Two observers analysed the video records to estimate the time spent for each PA (seconds and frequency).5. To calculate the energetic coefficients, the HP was regressed with the function of time spent to perform each PA allowing to estimate the energy cost for eating, drinking and stand activities, which were 0.607, 0.352 and 0.938 cal/kg-0.75/s, respectively.

Role of sex hormones in hypercapnia-induced activation of the locus coeruleus in female and male rats.

The locus coeruleus (LC) has been suggested as a CO2 chemoreceptor site in mammals. Most of the studies involving the role of the LC in hypercapnic ventilatory responses have been performed in males. Since ovarian steroids modulate the activity of LC neurons and females have a different respiratory response to CO2 than males, we evaluated the activity of LC noradrenergic neurons during normocapnia and hypercapnia in female and male rats with distinct sex hormone levels. Ovariectomized (OVX), estradiol (E2)-treated ovariectomized (OVX+E2) and female rats on the diestrous day of the estrous cycle were evaluated. Concurrently, males were investigated as gonad-intact, orchidectomized (ORX), testosterone (T)-treated ORX (ORX+T), and E2-treated ORX (ORX+E2). Activation of LC neurons was determined by double-label immunohistochemistry to c-Fos and tyrosine hydroxylase (TH). Hypercapnia induced by 7% CO2 increased the number of c-Fos/TH-immunoreactive (ir) neurons in the LC of all groups when compared to air exposure. Hypercapnia-induced c-Fos expression did not differ between diestrous females and intact male rats. In the OVX+E2 group, there was attenuation in the c-Fos expression during normocapnia compared with OVX rats, but CO2 responsiveness was not altered. Moreover, in ORX rats, neither T nor E2 treatments changed c-Fos expression in LC noradrenergic neurons. Thus, in female rats, E2 reduces activation of LC noradrenergic neurons, whereas in males, sex hormones do not influence the LC activity.

TRPV4 activates autonomic and behavioural warmth-defence responses in Wistar rats.

AIM: In this study, we aimed at investigating the involvement of the warmth-sensitive channel - TRPV4 (in vitro sensitive to temperatures in the range of approx. 24-34 °C) - on the thermoregulatory mechanisms in rats. METHODS: We treated rats with a chemical selective agonist (RN-1747) and two antagonists (RN-1734 and HC-067047) of the TRPV4 channel and measured core body temperature, metabolism, heat loss index and preferred ambient temperature. RESULTS: Our data revealed that chemical activation of TRPV4 channels by topical application of RN-1747 on the skin leads to hypothermia and this effect was blocked by the pre-treatment with the selective antagonist of this channel. Intracerebroventricular treatment with RN-1747 did not cause hypothermia, indicating that the observed response was indeed due to activation of TRPV4 channels in the periphery. Intravenous blockade of this channel with HC-067047 caused an increase in core body temperature at ambient temperature of 26 and 30 °C, but not at 22 and 32 °C. At 26 °C, HC-067047-induced hyperthermia was accompanied by increase in oxygen consumption (an index of thermogenesis), while chemical stimulation of TRPV4 increased tail heat loss, indicating that these two autonomic thermoeffectors in the rat are modulated through TRPV4 channels. Furthermore, rats chemically stimulated with TRPV4 agonist choose colder ambient temperatures and cold-seeking behaviour after thermal stimulation (28-31 °C) was inhibited by TRPV4 antagonist. CONCLUSION: Our results suggest, for the first time, that TRPV4 channel is involved in the recruitment of behavioural and autonomic warmth-defence responses to regulate core body temperature.

Participation of the dorsal periaqueductal grey matter in the hypoxic ventilatory response in unanaesthetized rats.

AIM: Although periaqueductal grey matter activation is known to elicit respiratory and cardiovascular responses, the role of this midbrain area in the compensatory responses to hypoxia is still unknown. To test the participation of the periaqueductal grey matter in cardiorespiratory and thermal responses to hypoxia in adult male Wistar rats, we performed a chemical lesion of the dorsolateral/dorsomedial or the ventrolateral/lateral periaqueductal grey matter using ibotenic acid. METHODS: Pulmonary ventilation, mean arterial pressure, heart rate and body temperature were measured in unanaesthetized rats during normoxic and hypoxic exposure (5, 15, 30 min, 7% O2). RESULTS: An ibotenic acid lesion of the dorsolateral/dorsomedial periaqueductal grey matter caused a higher increase in pulmonary ventilation (67.1%, 1730±282.5 mL kg(-1) min(-1)) compared to the Sham group (991.4±194 mL kg(-1) min(-1)) after 15 min in hypoxia, whereas for the ventrolateral/Lateral periaqueductal grey matter lesion, no differences were observed between groups. Mean arterial pressure, heart rate and body temperature were not affected by a dorsolateral/dorsomedial or ventrolateral/lateral periaqueductal grey matter lesion. CONCLUSION: Middle to caudal portions of the dorsolateral/dorsomedial periaqueductal grey matter neurones modulate the hypoxic ventilatory response, exerting an inhibitory modulation during low O2 situations. In addition, the middle to caudal portions of the dorsolateral/dorsomedial or ventrolateral/lateral periaqueductal grey matter do not appear to exert a tonic role on cardiovascular or thermal parameters during normoxic and hypoxic conditions.

Ionotropic but not metabotropic glutamatergic receptors in the locus coeruleus modulate the hypercapnic ventilatory response in unanaesthetized rats.

AIM: Central chemoreceptors are important to detect changes of CO2/H(+), and the Locus coeruleus (LC) is one of the many putative central chemoreceptor sites. Here, we studied the contribution of LC glutamatergic receptors on ventilatory, cardiovascular and thermal responses to hypercapnia. METHODS: To this end, we determined pulmonary ventilation (V(E)), body temperatures (T(b)), mean arterial pressure (MAP) and heart rate (HR) of male Wistar rats before and after unilateral microinjection of kynurenic acid (KY, an ionotropic glutamate receptor antagonist, 10 nmol/0.1 µL) or α-methyl-4-carboxyphenylglycine (MCPG, a metabotropic glutamate receptor antagonist, 10 nmol/0.1 µL) into the LC, followed by 60 min of air breathing or hypercapnia exposure (7% CO2). RESULTS: Ventilatory response to hypercapnia was higher in animals treated with KY intra-LC (1918.7 ± 275.4) compared with the control group (1057.8 ± 213.9, P < 0.01). However, the MCPG treatment within the LC had no effect on the hypercapnia-induced hyperpnea. The cardiovascular and thermal controls were not affected by hypercapnia or by the injection of KY and MCPG in the LC. CONCLUSION: These data suggest that glutamate acting on ionotropic, but not metabotropic, receptors in the LC exerts an inhibitory modulation of hypercapnia-induced hyperpnea.

Role of brain nitric oxide in the thermoregulation of broiler chicks.

Nitric oxide (NO) plays a key role in body temperature (Tb) regulation of mammals, acting on the brain to stimulate heat loss. Regarding birds, the putative participation of NO in the maintenance of Tb in thermoneutrality or during heat stress and the site of its action (periphery or brain) is unknown. Thus, we tested if NO participates in the maintenance of chicks' Tb in those conditions. We investigated the effect of intramuscular (im; 25, 50, 100mg/kg) or intracerebroventricular (icv; 22.5, 45, 90, 180 microg/animal) injections of the non selective NO synthase inhibitor L-NAME on Tb of 5-day-old chicks at thermoneutral zone (TNZ; 31-32 degrees C) and under heat stress (37 degrees C for 5-6h). We also verified plasma and diencephalic nitrite/nitrate levels in non-injected chicks under both conditions. At TNZ, 100mg/kg (im) or 45, 90, 180 microg (icv) of L-NAME decreased Tb. A significant correlation between Tb and diencephalic, but not plasma, nitrite/nitrate levels was observed. Heat stress-induced hyperthermia was inhibited by all tested doses of L-NAME (im and icv). Tb was correlated neither with plasma nor with diencephalic nitrite/nitrate levels during heat stress. These results indicate the involvement of brain NO in the maintenance of Tb of chicks, an opposite action of that observed in mammals, and may modulate hyperthermia.

Indomethacin impairs LPS-induced behavioral fever in toads.

We tested the hypothesis that PGs mediate lipopolysaccharide (LPS)-induced behavioral fever in the toad Bufo paracnemis. Measurements of preferred body temperature (T(b)) were performed with a thermal gradient. Toads were injected intraperitoneally with the cyclooxygenase inhibitor indomethacin (5 mg/kg), which inhibits PG biosynthesis, or its vehicle (Tris) followed 30 min later by LPS (0.2 and 2 mg/kg) into the lymph sac. LPS at the dose of 0.2 mg/kg caused a significant increase in T(b) from 7 to 10 h after injection, and then T(b) returned toward baseline values. LPS at the dose of 2 mg/kg produced a different pattern of response, with a longer latency to the onset of fever (10th h) and a longer duration (until the end of the experiment at the 15th h). Tris significantly attenuated the fever induced by LPS at 0.2 mg/kg, but not at 2 mg/kg. Moreover, indomethacin completely blocked the fever evoked by LPS (2 mg/kg). These results indicate that the behavioral fever induced by LPS in toads requires the activation of the COX pathway, suggesting that the involvement of PG in fever has an ancient phylogenetic history and that endogenous PGs raise the thermoregulatory set point to produce fever, because behavioral thermoregulation seems to be related to changes in the thermoregulatory set point.