Influence of soil hydric parameters on the winter cold hardiness of a burrowing beetle, Leptinotarsa decemlineata (Say)

This investigation examined the influence of soil moisture and associated parameters on the cold hardiness of the Colorado potato beetle (Leptinotarsa decemlineata Say), a temperate-zone species that overwinters in terrestrial burrows. The body mass and water content of adult beetles kept in sand at 4 °C varied over a 16-week period of diapause according to the substratum's moisture content. Changes in body water content, in turn, influenced the crystallization temperature (range −3.3 to −18.4 °C; n = 417), indicating that environmental moisture indirectly determined supercooling capacity, a measure of physiological cold hardiness. Beetles held in dry sand readily tolerated a 24-h exposure to temperatures ranging from 0° to −5 °C, but those chilled in sand containing as little as 1.7% water (dry mass) had elevated mortality. Thus, burrowing in dry soils not only promotes supercooling via its effect on water balance, but may also inhibit inoculative freezing. Mortality of beetles exposed to −5 °C for 24 h was lower in substrates composed of sand, clay and/or peat (36–52%) than in pure silica sand (78%) having an identical water content (17.0% dry mass). In addition to moisture, the texture, structure, water potential, and other physico-chemical attributes of soil may strongly influence the cold hardiness and overwintering survival of burrowing insects. Accepted: 10 September 1996     

Phenotypic variation in seasonal adjustments of testis size, body weight, and food intake in deer mice: role of pineal function and ambient temperature

We investigated pineal function as well as reproductive and energetic characteristics in male deer mice (Peromyscus maniculatus) that differentially respond to short photoperiod with full, partial or no gonadal regression. In mice at both high (23 °C) and low temperature (1 °C), these phenotypic differences in reproductive responses to short days were not reflected by differences in urinary excretion of 6-sulphatoxy-melatonin, the main metabolite of pineal melatonin. Neither duration nor amplitude or phase-angle of nocturnal peaks in 6-sulphatoxymelatonin significantly differed between reproductive phenotypes at either temperature. Differences in testis size were, however, associated with different energy requirements. In gonadally regressed males only, food intake and body weight were significantly (P < 0.01) reduced by up to 29% and 13% respectively. Chronic cold exposure (5 °C) had no effect on the proportion of males undergoing testicular regression under short days, but caused a general elevation in body weights among all mice (P < 0.05). Phenotypic differences in body weight and food intake were maintained in the cold. Together, these results suggest that within-population variation of reproductive responses in male deer mice is based on post-pineal differences in the regulation of gonadal function, and that phenotypic characteristics in reproductive and energetic responses to short days are largely unaffected by ambient temperature. Accepted: 2 October 1995     

Temperature induced anaerobiosis in two populations of the polychaete worm Arenicola marina (L.)

Temperature dependent changes in the mode of energy metabolism and in acid-base status were studied in the range from −1.7 to 26 °C in two populations of Arenicola marina collected in summer as well as in winter from intertidal flats of the North Sea (boreal) and the White Sea (subpolar). Extreme temperatures led to an accumulation of anaerobic end products, indicating the existence of both a low and a high critical temperature, beyond which anaerobic metabolism becomes involved in energy production. In summer animals from the North Sea the high critical temperature was found at temperatures above 20 °C, and the low critical temperature below 5 °C. Latitudinal or seasonal cold adaptation lead to a more or less parallel shift of both high and low critical temperature values to lower values. Between critical temperatures intracellular pH declined with rising temperature. Slopes varied between −0.012 and −0.022 pH- units/°C. In summer animals from the North Sea, the slope was slightly less than in White Sea animals, but differences appeared independent of the season. However, slopes were no longer linear beyond critical temperatures. A drop in intracellular pH at low temperatures coincided with the accumulation of volatile fatty acids in the body wall tissue of North Sea animals. A failure of active pH_i adjustment is held responsible for the reduced ΔpH_i/ΔT at temperatures above the high critical temperature. Extracellular pH was kept constant over the whole temperature range investigated. The ability of North Sea animals to adapt to temperatures beyond the critical temperature is poor compared to White Sea specimens. The larger range of temperature fluctuations at the White Sea is seen as a reason for the higher adaptational capacity of the subpolar animals. A hypothesis is proposed that among other mechanisms critical temperature values are set by an adjustment of mitochondrial density and thus, aerobic capacity. Accepted: 20 August 1996     

Second messenger and cAMP-dependent protein kinase responses to dehydration and anoxia stresses in frogs

The effects of whole body dehydration (up to 40% of total body water lost) or anoxia exposure (up to 2 days under N₂ gas) at 5 °C on tissue levels of adenosine 3′–5′ cyclic monophosphate (cAMP) and the percentage of cAMP-dependent protein kinase present as the free catalytic subunit (PKAc), as well as the levels of the protein kinase C (PKC) second messenger, inositol 1,4,5-trisphosphate (IP₃), were assessed in two anurans, the freeze-tolerant wood frog, Rana sylvatica, and the freeze-intolerant leopard frog, Rana pipiens. Dehydration of wood frogs resulted in a rapid elevation of liver cAMP and PKAc; cAMP was 3.4-fold greater than control values in animals that had lost 5% of total body water, whereas PKAc was elevated threefold in 20% dehydrated frogs. These results indicate protein kinase A mediation of the liver glycogenolysis and hyperglycemia that is induced by dehydration in this species. Skeletal muscle PKAc content also rose with dehydration but neither cAMP nor PKAc was affected by dehydration in leopard frog tissues. Anoxia exposure had different effects on signal transduction systems. PKAc was elevated after 1 h anoxia in R. sylvatica brain and was sustained over time but the enzyme was unaffected in other organs; by contrast, R. pipiens showed variable responses by PKAc to anoxia in three organs. Both species showed rapid (within 30 min) and large (3 to 7.8-fold) increases in IP₃ in liver of anoxic frogs that decreased slowly with continued anoxia. IP₃ also increased quickly in heart of anoxia-exposed wood frogs. This suggests that PKC may mediate various metabolic adjustments that promote hypoxia/anoxia resistance such as coordinating metabolic rate depression. A progressive rise in liver IP₃ during dehydration in wood frogs (reaching fourfold higher than controls in 40% dehydrated animals) may also mediate similar hypoxia resistance adaptations under this stress since anurans experience progressive hypoxia due to increased blood viscosity when water loss reaches high values. The patterns of second messenger and PKAc changes in wood frog liver during dehydration closely parallel the changes seen in these same parameters during natural freezing suggesting that the freeze tolerance of selected terrestrially hibernating anurans may have evolved out of various anuran mechanisms of dehydration resistance. Accepted: 2 January 1997     

Cold tolerance and dehydration in Enchytraeidae from Svalbard

When cooled in contact with moisture, eight species of arctic Enchytraeidae from Svalbard were killed by freezing within minutes or hours at −3 and −5 °C; an exception was Enchytraeus kincaidi which survived for up to 2 days. When the temperature approached 0 °C the enchytraeids apparently tried to escape from the moist soil. The supercooling capacity of the enchytraeids was relatively low, with mean supercooling points of −5 to −8 °C. In contrast, specimens of several species were extracted from soil cores that had been frozen in their intact state at −15 °C for up to 71 days. Compared to freezing in a moist environment, higher survival rates were obtained during cooling at freezing temperatures in dry soil. Survival was recorded in species kept at −3 °C for up to 35 days, and in some species kept at −6 °C for up to 17 days. Slow warming greatly increased survival rates at −6 °C . The results strongly suggest that arctic enchytraeids avoid freezing by dehydration at subzero temperatures. In agreement with this, weight losses of up to ca. 42% of fresh weight were recorded in Mesenchytraeus spp. and of up to 55% in Enchytraeus kincaidi at water vapour pressures above ice at −3 to −6 °C. All specimens survived dehydration under these conditions. Accepted: 12 December 1997     

Ontogeny of deep-body cold sensitivity in Pekin ducklings Anas platyrhynchos

The ontogeny of deep-body cold sensitivity was studied in 1 to 12 days old Pekin ducklings Anas platyrhynchos. Deep-body cold sensitivity was determined by means of thermodes implanted in the abdominal cavity. The thermodes were perfused with cold water for 15-min periods to lower the core temperature. Cooling of the body core elicited increases in metabolic rate and vasoconstrictions in the legs of all the ducklings. From the changes induced in metabolic rate and core temperature, deep-body cold sensitivity values of between −5.17 and −6.36 W · kg⁻¹ · °C⁻¹, were estimated. These values, which are in the range of those reported previously for adult Pekin ducks, did not change with age, and it is concluded that deep-body cold sensitivity is fully developed at hatching. Our next aim was to investigate whether the autonomic responses elicited by exposure of ducklings to cold ambient conditions could be explained by temperature changes within the body core. During cold exposure, the increase in metabolic rate was not accompanied by a concomitant decrease in core temperature. On the contrary, deep-body temperature increased slightly during the initial phase of cold exposure. The ducklings attained a metabolic rate amounting to 85–90% of their peak metabolic rate before the core temperature fell below the regulated level measured at thermoneutrality. Thus, despite the findings that Pekin ducklings have a highly-developed deep-body cold sensitivity, their metabolic cold defence under natural conditions seems to be mediated primarily by peripheral thermoreceptors. Accepted: 7 January 1997     

Blood flow in the carotid artery during breath-holding in relation to diving bradycardia

The present study investigated the mechanism of diving bradycardia. A group of 14 healthy untrained male subjects were examined during breath-holding either out of the water (30–33°C), in head-out immersion, or in whole-body submersion (27–29°C) in a diving pool. Blood velocity, blood volume flow in the carotid artery, diastolic blood pressure and electrocardiogram were measured and recorded during the experiments. The peak blood velocity increased by 13.6% (P < 0.01) and R-wave amplitude increased by 57.1% (P < 0.005) when the subjects entered water from air. End-diastolic blood velocity in the carotid artery increased significantly during breath-holding, e.g. increased from 0.20 (SD 0.02) m · s⁻¹ at rest to 0.33 (SD 0.04) m · s⁻¹ (P < 0.001) at 50.0 s in breath-hold submersion to a 2.0-m depth. Blood volume flow in the carotid artery increased by 26.6% (P < 0.05) at 30 s and 36.6% (P < 0.001) at 40 s in breath-hold submersion to a 2.0-m depth. Diastolic blood pressure increased by 15.4% (P < 0.01) at 60 s during breath-holding in head-out immersion. Blood volume flow, and diastolic blood pressure increased significantly more and faster during breath-holding in submersion than out of the water. There was a good negative correlation with the heart rate: the root mean square correlation coefficient r was 0.73 (P < 0.001). It was concluded that an increased accumulation of blood in the aorta and arteries at end-diastole and decreased venous return, caused by an increase in systemic peripheral resistance during breath-holding, underlies diving bradycardia. Accepted: 22 November 1996     

Temperature regulation and metabolism of an Australian bat, Chalinolobus gouldii (Chiroptera: Vespertilionidae) when euthermic and torpid

The thermal and metabolic physiology of Chalinolobus gouldii, an Australian vespertilionid bat, was studied in the laboratory using flow-through respirometry. Chalinolobus gouldii exhibits a clear pattern of euthermic thermoregulation, typical of endotherms with respect to body temperature and rate of oxygen consumption. The basal metabolic rate of euthermic Chalinolobus gouldii is approximately 86% of that predicted for a 17.5-g mammal and falls into the range of mass-specific basal metabolic rates ascribed to vespertilionid bats. However, like most vespertilionid bats, Chalinolobus gouldii displays extreme thermolability. It is able to enter into torpor and spontaneously arouse at ambient temperatures as low as 5 °C. Torpid bats thermoconform at moderate ambient temperature, with body temperature ≈ ambient temperature, and have a low rate of oxygen consumption determined primarily by Q ₁₀ effects. At low ambient temperature (< 10 °C), torpid C. gouldii begin to regulate their body temperature by increased metabolic heat production; they tend to maintain a higher body temperature at low ambient temperature than do many northern hemisphere hibernating bats. Use of torpor leads to significant energy savings. The evaporative water loss of euthermic bats is relatively high, which seems unusual for a bat whose range includes extremely arid areas of Australia, and is reduced during torpor. The thermal conductance of euthermic C. gouldii is less than that predicted for a mammal of its size. The thermal conductance is considerably lower for torpid bats at intermediate body temperature and ambient temperature, but increases to euthermic values for torpid bats when thermoregulating at low ambient temperature. Accepted: 22 August 1996     

The influence of either no fluid or carbohydrate-electrolyte fluid ingestion and the environment (thermoneutral versus hot and humid) on running economy after prolonged, high-intensity exercise

This study investigated the effects on running economy (RE) of ingesting either no fluid or an electrolyte solution with or without 6% carbohydrate (counterbalanced design) during 60-min running bouts at 80% maximal oxygen consumption (V˙O_2max). Tests were undertaken in either a thermoneutral (22–23°C; 56–62% relative humidity, RH) or a hot and humid natural environment (Singapore: 25–35°C; 66–77% RH). The subjects were 15 young adult male Singaporeans [V˙O_2max = 55.5 (4.4 SD) ml kg⁻¹ min⁻¹]. The RE was measured at 3 m s⁻¹ [65 (6)% V˙O_2max] before (RE1) and after each prolonged run (RE2). Fluids were administered every 2 min, at an individual rate determined from prior tests, to maintain body mass (group mean = 17.4 ml min⁻¹). The V˙O₂ during RE2 was higher (P < 0.05) than that during the RE1 test for all treatments, with no differences between treatments (ANOVA). The mean increase in V˙O₂ from RE1 to RE2 ranged from 3.4 to 4.7 ml kg⁻¹ min⁻¹ across treatments. In conclusion, the deterioration in RE at 3 m s⁻¹ (65% V˙O_2max) after 60 min of running at 80% V˙O_2max appears to occur independently of whether fluid is ingested and regardless of whether the fluid contains carbohydrates or electrolytes, in both a thermoneutral and in a hot, humid environment. Accepted: 30 October 1997     

Cold induced vasodilatation and cardiovascular responses in humans during cold water immersion of various upper limb areas

To study the physiological responses induced by immersing in cold water various areas of the upper limb, 20 subjects immersed either the index finger (T1), hand (T2) or forearm and hand (T3) for 30 min in 5°C water followed by a 15-min recovery period. Skin temperature of the index finger, skin blood flow (Q_sk) measured by laser Doppler flowmetry, as well as heart rate (HR) and mean arterial blood pressure (ˉBP_a) were all monitored during the test. Cutaneous vascular conductance (CVC) was calculated as Q_sk / ˉBP_a. Cold induced vasodilatation (CIVD) indices were calculated from index finger skin temperature and CVC time courses. The results showed that no differences in temperature, CVC or cardiovascular changes were observed between T2 and T3. During T1, CIVD appeared earlier compared to T2 and T3 [5.90 (SEM 0.32) min in T1 vs 7.95 (SEM 0.86) min in T2 and 9.26 (SEM 0.78) min in T3, P < 0.01]. The HR was unchanged in T1 whereas it increased significantly at the beginning of T2 and T3 [+13 (SEM 2) beats · min⁻¹ in T2 and +15 (SEM 3) beats · min⁻¹ in T3, P < 0.01] and then decreased at the end of the immersion [−12 (SEM 3) beats · min⁻¹ in T2, and −15 (SEM 3) beats · min⁻¹ in T3, P < 0.01]. Moreover, ˉBP_aincreased at the beginning of T1 but was lower than in T2 and T3 [+9.3 (SEM 2.5) mmHg in T1, P < 0.05;  +20.6 (SEM 2.6) mmHg and 26.5 (SEM 2.8) mmHg in T2 and T3, respectively, P < 0.01]. The rewarming during recovery was faster and higher in T1 compared to T2 and T3. These results showed that general and local physiological responses observed during an upper limb cold water test differed according to the area immersed. Index finger cooling led to earlier and faster CIVD without significant cardiovascular changes, whereas hand or forearm immersion led to a delayed and slower CIVD with a bradycardia at the end of the test. Accepted: 26 November 1996     

Effect of masking the parietal eye on the diurnal activity and body temperature of two sympatric species of monitor lizards, Varanus s. salvator and Varanus b. nebulosus

 Five water monitor lizards, Varanus salvator salvator, and four clouded monitor lizards, Varanus bengalensis nebulosus, were caught on Tioman island in Malaysia. A radio-thermistor transmitter was implanted into the buccal cavity of each animal, and they were released into an enclosure measuring 5.5 × 6.5 metres. The lizards were observed for 9 and 8 days, respectively, before and after the parietal eye was covered with aluminium foil. With uncovered parietal eye, both species showed a clear diurnal rhythm, being active only during day time. After covering the parietal eye, the mean locomotor activity of five V. s. salvator decreased from 791 to 107 min · day^–1 but remained unchanged around 850 min · day^–1 for V. b. nebulosus. The mean duration of locomotor activity decreased in V. s. salvator and V. b. nebulosus after the parietal eye was covered, but V. b. nebulosus maintained its locomotor activity by increasing the number of locomotor bouts. The water monitor spent very little time on thermoregulation. Its body temperature ranged between 26.3 and 28.4 °C, which decreased after the parietal eye was covered. The clouded monitor thermoregulated around 28.8–36.0 °C, which remained unchanged after the parietal eye was covered. In both species, there was a strong correlation between body temperature and ambient temperature. Behavioural abnormalities were recorded among V. s. salvator with covered parietal eye. They were often observed to be active by night and often slept outside a burrow. The circadian rhythm of V. b. nebulosus appeared unaffected by shielding of its parietal eye. Captivity combined with shielded parietal eye induced agonistic behaviour in both species. Accepted: 11 September 1996     

Rhythmic components of photoperiodic time measurement in the pitcher-plant mosquito, Wyeomyia smithii

Photoperiodic time measurement regulating larval diapause in the pitcher-plant mosquito, Wyeomyia smithii, varies in a close relationship with latitude. The critical photoperiod mediating the maintenance and termination of diapause is positively correlated with latitude (r ² = 0.977) among six populations from southern (30–31° N), intermediate (40° N), and northern (46–49° N) latitudes in North America. The developmental response to unnaturally short and to unnaturally long photoperiods declines with increasing latitude, so that longer critical photoperiods are associated with a downward rather than a lateral shift in the photoperiodic response curve. Exotic light and dark cycles of varying period (T) with a short (10 h) photophase and a scotophase ranging from 14 (T = 24) to 62 (T = 72) h, reveal two geographic patterns: a decline in perturbability of the photoperiodic clock with increasing latitude, and no change with latitude in the 21-h period of rising and falling development with increasing T. These results show (1) that there is a rhythmic component to photoperiodic time measurement in W. smithii, (2) that the period of this rhythm is about 21 h in all populations, and (3) that more northern populations show decreasing responsiveness to photoperiod and increasing stability against perturbation by exotic period lengths (T > 24). Previous studies on W.␣smithii indicate that this single temperate species of a tropical and subtropical genus has evolved from south to north. We therefore conclude that the evolution of increasing critical photoperiod in W. smithii during its adaptive radiation into North America has more likely involved the amplitude and not the period of the underlying circadian pacemaker. Received: 22 July 1996 / Accepted: 30 September 1996     

Fatty acid composition and ultrastructure of photoreceptive membranes in the crayfish Procambarus clarkii under conditions of thermal and photic stress

The ultrastructural state of the crayfish visual membrane is correlated with its fatty acid composition during times of photic and thermal stress and the period over which the dynamic events occur is investigated. Crayfish kept at 4 °C under constant darkness contain in their rhabdoms significantly increased amounts of unsaturated fatty acids such as 16:1, 18:1, 20:5, and 22:6 compared with individuals kept at 25 °C. The ratio of unsaturated/saturated fatty acids (UFA/SFA-ratio) amounts to 2.17 in the cold-water- and 1.46 in the warm water-acclimated animals. The visual membranes of crayfish suddenly transferred from 4 °C to 25 °C exhibited ultrastructural modifications such as membrane collapse and disappearance of microvillar dense␣core-filaments most clearly 3 h post-transfer. Parallel to the structural changes a significant increase in fatty acid 18:0 was observed, while the amounts of 16:1 and 20:1 decreased. When 4 °C, dark-adapted crayfish were exposed to light alone and not a temperature increase, only fatty acid 22:6 showed a significant reduction to 10% of its pre-experimental level within 2 h of exposure. Thereafter, it slowly increased again. In cold water-acclimated crayfish that had been exposed to light of 5000 lx for 3␣weeks no significant change of the UFA/SFA ratio was observed, although fatty acid species 18:0, 20:4, and 20:5 had increased at the expense of fatty acids 14:0, 16:0, 16:1, 18:1, 20:1, and 22:6. The total amount of fatty acids, however, had become significantly smaller (from 0.058 ng g⁻¹ body weight in the dark-adapted to 0.048 ng g⁻¹ in the light-adapted crayfish). Morphologically the rhabdom volume had decreased by approx. 20%, but ultrastructurally rhabdom microvilli remained almost unchanged. The amount of peroxidized lipids in the retina following irradiation with bright white light in the cold-adapted crayfish fell during the first 2 h of exposure from 0.4 nmol g⁻¹ to 0.32 nmol g⁻¹, but after 12 h of exposure had reached a level of 0.48 nmol g⁻¹. Greatest structural abnormalities to the visual membranes occurred when dark-adapted, cold-acclimated crayfish were suddenly subjected to bright light and an increase in water temperature. Under such conditions the microvillar arrangement was disrupted and membrane collapse and disappearance of core-filaments were apparent. Our results provide evidence that the fatty acid composition of the membranes determines to a considerable extent the structural integrity of the photoreceptor, but that it is too simplistic a model to think that peroxidation of membrane lipids alone is responsible for the disintegration of the photoreceptive membranes in the crayfish eye following exposure to bright light. Accepted: 4 July 1996     

A wat1 mutant of fission yeast is defective in cell morphology

The organization of the actin cytoskeleton plays an integral role in cell morphogenesis of all eukaryotes. We have isolated a temperature-sensitive mutant in Schizosaccharomyces pombe, wat1-1, in which acting patches are delocalized, resulting in an elliptically shaped cell phenotype. Molecular cloning and DNA sequencing of wat1 ⁺ showed that the gene encodes a 314 residue protein containing WD-40 repeats. Cells lacking wat1 ⁺ are slow growing but viable at 25° C and temperature-sensitive for growth above 33° C. At restrictive temperature, wat1-d strains are phenotypically indistinguishable from wat1-1. When combined with a deletion for the wat1 ⁺ gene, cdc mutants failed to elongate at restrictive temperature and exhibited alterations in actin patch localization. This analysis suggests that wat1 ⁺ is required directly or indirectly for polarized cell growth in S. pombe. Wat1p and a functional, epitope-tagged, version of Wat1p can be overproduced without inducing alterations in cell morphology. Received: 18 September 1996 / Accepted: 22 October 1996     

Effects of hydration state on hormonal and renal responses during moderate exercise in the heat

The effects of hydromineral hormones and catecholamines on renal concentrating ability at different hydration states were examined in five male volunteers while they performed three trials. Each of these trials comprised a 60-min exercise bout on a treadmill (at 50% of maximal oxygen uptake) in a warm environment (dry bulb temperature, 35°C; relative humidity, 20–30%). In one session, subjects were euhydrated before exercise (C). In the two other sessions, after thermal dehydration (loss of 3% body mass) which markedly reduced plasma volume (PV) and increased plasma osmolality (osm_pl), the subjects exercised either not rehydrated (Dh) or rehydrated (Rh) by drinking 600 ml of mineral water before and 40 min after the onset of exercise. During exercise in the Dh compared to C state, plasma renin, aldosterone, arginine vasopressin (AVP), noradrenaline and adrenaline concentrations were increased (P < 0.05). A reduction in creatinine clearance and urine flow was also observed (P < 0.05) together with a decrease in urine osmolality, osmolar clearance and sodium excretion, while free water clearance increased (P < 0.05). However, compared to Dh, Rh partially restored PV and osm_pl and induced a marked reduction in the time courses of both the plasma AVP and catecholamine responses (P < 0.05). Values for renal water and electrolyte excretion were intermediate between those of Dh and C. Plasma atrial natriuretic peptide presented similar changes whatever the hydration state. These results demonstrate that during moderate exercise in the heat, renal concentrating ability is paradoxically reduced by prior dehydration in spite of high plasma AVP levels, and might be the result of marked activation of the sympatho-adrenal system. Rehydration, by reducing this activation, could partially restore the renal concentrating ability despite the lowered plasma AVP. Accepted: 23 April 1997     

Cold hardiness in Entomobrya nivalis (Collembola, Entomobryidae): annual cycle of polyols and antifreeze proteins, and antifreeze triggering by temperature and photoperiod

In the Swiss Prealps Entomobrya nivalis hibernates in an inactive state, hidden under bark flakes on spruce. For freeze avoidance it relies on thermal hysteresis proteins (THPs) and polyols (mainly ribitol, with small amounts arabitol and threitol). Polyols are present only during the inactive state, THPs additionally protect during the transition phase in spring and autumn, when animals are still active but frosts may occur. Peak values were recorded in February/March for THPs (3.5 °C hysteresis between melting and freezing point) and for polyols (26 μg mg⁻¹ FW; hemolymph osmolality 680 mosmol l⁻¹). E. nivalis is able to control its hemolymph osmolality independently of body water content. Mean osmolality in summer was 350– 440 mosmol l⁻¹, in winter it was elevated to 650 mosmol l⁻¹, due to a synthesis mainly of ribitol. Body water content varied between 1.8 and 3.3 mg H₂O mg⁻¹ DW, depending on humidity conditions. Experiments on triggering of antifreeze synthesis showed the action of temperature and photoperiod as cues, but there was also evidence for an endogenous rhythm. No clear correlation between antifreeze concentration and supercooling ability could be established, suggesting that gut content or other parameters also play an inportant role. Accepted: 18 November 1995     

Short latency vestibular evoked potentials in the Japanese quail (Coturnix coturnix japonica)

Short-latency vestibular-evoked potentials to pulsed linear acceleration were characterized in the quail. Responses occurred within 8 ms following the onset of stimuli and were composed of a series of positive and negative peaks. The latencies and amplitudes of the first four peaks were quantitatively characterized. Mean latencies at 1.0 g ms⁻¹ ranged from 1265 ± 208 μs (P1, N = 18) to 4802 ± 441 μs (N4, N = 13). Amplitudes ranged from 3.72 ± 1.51 μV (P1/N1, N = 18) to 1.49 ± 0.77 μV (P3/N3, N = 16). Latency-intensity (LI) slopes ranged from −38.7 ± 7.3 μs dB⁻¹ (P1, N = 18) to −71.6 ± 21.9 μs dB⁻¹ (N3, N = 15) and amplitude-intensity (AI) slopes ranged from 0.20 ± 0.08 μV dB⁻¹ (P1/N1, N = 18) to 0.07 ± 0.04 μV dB⁻¹ (P3/N3, N = 11). The mean response threshold across all animals was −21.83 ± 3.34 dB re: 1.0 g ms⁻¹ (N = 18). Responses remained after cochlear extirpation showing that they could not depend critically on cochlear activity. Responses were eliminated by destruction of the vestibular end organs, thus showing that responses depended critically and specifically on the vestibular system. The results demonstrate that the responses are vestibular and the findings provide a scientific basis for using vestibular responses to evaluate vestibular function through ontogeny and senescence in the quail. Accepted: 18 January 1997     

c-fos expression in the putative avian suprachiasmatic nucleus

c-fos induction was investigated as a potential component in the avian photic entrainment pathway and as a possible means of locating the central pacemaker in birds. In both quail (Coturnix coturnix japonica) and starlings (Sturnus vulgaris) exposure to 1 h of light induced Fos-lir in the visual suprachiasmatic nucleus but not in the medial suprachiasmatic nucleus. However, the degree of c-fos induction in the visual suprachiasmatic nucleus was similar at different circadian times despite the fact that the light pulses caused differential phase shifts in the locomotor rhythm. For golden hamsters the same experiment resulted in significantly different levels of Fos-lir in the suprachiasmatic nucleus, as well as different phase shifts. Starlings and hamsters were also entrained to T-cycles that caused a large daily phase shift (T = 21.5 h in starlings, T = 22.67 hours in hamsters), or no daily phase shift (T = free running period). No difference in the induced levels of Fos-lir in the visual suprachiasmatic nucleus region was observed between the two groups of starlings, but in hamsters there were significantly different levels of Fos-lir in the suprachiasmatic nucleus between the two groups. Accepted: 15 November 1996