Circadian rhythm of behavioural responsiveness to carbon dioxide in the blood-sucking bug Triatoma infestans (Heteroptera: Reduviidae)

The temporal modulation of the behavioural response to carbon dioxide and its chronobiological basis were investigated in larvae of Triatoma infestans. We analysed the orientation towards CO(2) of insects kept under three different illumination regimes: (1) 12 h light/12 h darkness cycles (L/D), (2) constant darkness (D/D) and (3) constant light (L/L). When maintained under L/D conditions, insects exhibited an oriented response towards airstreams added with 1500 ppm of CO(2) during the first hours of the scotophase only. Bugs maintained under D/D also showed a positive orientation response towards CO(2) during the first hours of the subjective night, while bugs kept under L/L did not show a rhythmic oriented behaviour. Thus, T. infestans displayed a daily rhythm of orientation towards CO(2) (i.e. a potential food source) only at the beginning of the scotophase. The persistence of the rhythm under constant darkness reveals the existence of an endogenous circadian control of this behaviour.     

Daily rhythm of aggregation in the haematophagous bug Triatoma infestans (Heteroptera: Reduviidae)

Triatomine bugs show a temporal modulation of many activities. Here, we analyse the daily modulation of the aggregation behaviour of Triatoma infestans larvae and its chronobiological basis. In the laboratory, groups of six bugs were released over an experimental arena during six consecutive days, where their aggregation behaviour was quantified every hour. When submitted to a 12/12 h photoperiod (L/D), the larvae of T. infestans exhibited a cyclic pattern of aggregation with a 24 h period, evincing the existence of a daily rhythm of aggregation in this species. Bugs exhibited the maximum aggregation tendency at the end of the scotophase (7:00 h), moment in which they naturally search for refuges. The minimum aggregation (i.e. maximal dispersion) was observed during the last part of the photophase and beginning of the scotophase (15:00 to 1:00 h). This cyclic pattern disappeared when constant conditions of illumination (L/L) or darkness (D/D) were imposed to the bugs, suggesting the absence of an endogenous circadian control of this behaviour. Insects submitted to L/L and D/D photoperiods presented lower global levels of aggregation than those submitted to L/D conditions. The lack of an endogenous control and the relevance of light cycles as a synchronization signal are discussed as the temporal modulation of this behaviour might play an important role in the nocturnal habits of this species.     

Circadian activity rhythms and sensitivity to noise in the Mongolian gerbil (Meriones unguiculatus)

Since consistent data on endogenous circadian rhythms of Mongolian gerbils are not available, the main aim of our study was to identify suitable conditions to receive stable and reproducible free-running rhythms of activity under different light intensities. Another objective was to determine the role of social cues as an exogenous zeitgeber in the absence of a light-dark (LD) cycle. We performed two long-term sets of experiments with adult male gerbils kept in climatic chambers under various photoperiods of at least 30 days each. In all cases, the time of lights on in the chambers differed from the daily starting hour of work in the animal house. Always, two animals per chamber were kept separately in cages with a running wheel while their activity was monitored continuously. During the first set, only three of eight animals developed intra- and interindividual variable free-running rhythms. The activity patterns seemed to be influenced by human activities outside, indicating high sensitivity to external factors. Subsequently, we damped the chambers and the room and restricted access to the room. In the following noise-reduced set, all gerbils developed comparable free-running rhythms of activity. We determined the mean of the free-running period tau, the activity-rest relationship alpha/theta and the amount of running wheel activity per day: tau = 23.7h +/- 0.08h under low light (5 lux) and 25.5h +/- 0.19h under high light intensities (450 lux); alpha/theta = 0.53 +/- 0.08 under 5 lux and 0.34 +/- 0.04 under 450 lux. The amount of daily activity was 12 times as high under 5 lux as under 450 lux. There was no indication that the two animals in one chamber socially synchronized each other. In conclusion, the pronounced rhythm changes in accordance with Aschoff's theory support the view that gerbils are mainly nocturnal animals.     

The role of antennae in the thermopreference and biting response of haematophagous bugs

Insects sense thermal cues mainly through thermoreceptors located in the antenna. To analyse the impact of antennectomy on the thermal behaviour of the haematophagous bug Triatomainfestans, we studied the distribution of intact and antennectomised bugs in an experimental arena where a temperature gradient was established, as well as the biting response of insects with and without antennae to objects at the temperature of a potential host. Antennectomy did not abolish thermopreference, but modified the temperature at which the insects preferred to stay. In the arena, antennectomised insects chose to remain at a higher temperature (ca. 3 °C higher in unfed bugs), and exhibited a larger dispersion around that preferred temperature, than intact bugs. In addition, ablated insects temporarily lost their ability to bite an object at the temperature of a potential host, but that ability was gradually recovered after the fifth day post-antennectomy. Results presented here show that thermoreceptors other than those located on the antennae can also guide thermal behaviours. We conclude that the function of antennal thermoreceptors can be taken over by other receptors located in different regions of the body. Those receptors have a different sensitivity and confer the insects with a different responsiveness.     

Circadian regulation of bioluminescence in the prey-luring glowworm, Arachnocampa flava

The glowworms of New Zealand and Australia are bioluminescent fly larvae that generate light to attract prey into their webs. Some species inhabit the constant darkness of caves as well as the dim, natural photophase of rain-forests. Given the diversity of light regimens experienced by glowworms in their natural environment, true circadian rhythmicity of light output could be present. Consequently the light emission characteristics of the Australian subtropical species Arachnocampa flava, both in their natural rainforest habitat and in artificial conditions in the laboratory, were established. Larvae were taken from rainforest and kept alive in individual containers. When placed in constant darkness (DD) in the laboratory they maintained free-running, cyclical light output for at least 28 days, indicating that light output is regulated by an endogenous rhythm. The characteristics of the light emission changed in DD: individuals showed an increase in the time spent glowing per day and a reduction in the maximum light output. Most individuals show a free-running period greater than 24 h. Manipulation of the photophase and exposure to skeleton photoperiods showed that light acts as both a masking and an entraining agent and suggests that the underlying circadian rhythm is sinusoidal in the absence of light-based masking. Manipulation of thermoperiod in DD showed that temperature cycles are an alternative entraining agent. Exposure to a period of daily feeding in DD failed to entrain the rhythm in the laboratory. The endogenous regulation of luminescence poses questions about periodicity and synchronization of bioluminescence in cave glowworms.     

The effect of temperature on the behaviour and development of Triatoma brasiliensis

Abstract. The effects of temperature on the development of early stages and the thermopreference of nymphs and adults were analysed in the haematophagous bug Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae). Egg hatching, mortality of nymphs, feeding and moulting success of the early stages of T. brasiliensis were all affected by temperature. While high rates of egg hatching were observed between 25 and 27 °C, no hatching occurred at 12, 19 and 38 °C. The mortality of first‐instar nymphs was highest at 38 °C, at which no insects survived after 10 days of exposure. Feeding success was only affected at the lowest temperature (12 °C). No ecdysis was observed in the groups exposed to 12, 19 and 21 °C. Recently fed fourth‐instar nymphs preferred to stay at a temperature of approximately 30 °C. The preferred temperature began to decline gradually to approximately 27 °C during ecdysis, reaching 26 °C at 30 days after ecdysis. After a second blood meal, the insects' preferred temperature was again approximately 30 °C. The thermopreference pattern of females was similar to that of nymphs. Nymphs and females showed a daily fluctuation in their preferred temperature, moving towards higher values at the beginning of the dark phase, and choosing lower ones after this time interval, at which they remained until the end of the light phase. The females laid their eggs in all sectors of the arena, although the largest numbers of eggs were found between 28 and 32 °C.     

Circadian rhythm of spontaneous locomotor activity in the bed bug, Cimex lectularius L.

Bed bugs must avoid detection when finding hosts and returning to hidden harborages. Their stealthy habits include foraging when hosts are asleep. Characteristics of spontaneous locomotor activity rhythm of bed bugs with different feeding histories were studied. In the absence of host stimuli, adults and nymphs were much more active in the dark than in the light. The onset of activity in the scotophase commenced soon after lights-off. The free-running period (τ) for all stages was longer in continuous darkness (DD) than in continuous light (LL). The lengthening of τ in DD is an exception for the circadian rule that predicts the opposite in nocturnal animals. Activity in all stages was entrained to reverse L:D regimes within four cycles. Short-term starved adults moved more frequently than recently fed adults. While bed bugs can survive for a year or more without a blood meal, we observed a reduction in activity in insects held for five weeks without food. We suggest that bed bugs make a transition to host-stimulus dependent searching when host presence is not predictable. Such a strategy would enable bed bugs to maximize reproduction when resources are abundant and save energy when resources are scarce.     

Multi-oscillatory control of eclosion and oviposition rhythms in Drosophila melanogaster: evidence from limits of entrainment studies

The eclosion and oviposition rhythms of flies from a population of Drosophila melanogaster maintained under constant conditions of the laboratory were assayed under constant light (LL), constant darkness (DD), and light/dark (LD) cycles of 10:10h (T20), 12:12h (T24), and 14:14h (T28). The mean (+/- 95% confidence interval; CI) free-running period (tau) of the oviposition rhythm was 26.34 +/- 1.04h and 24.50 +/- 1.77h in DD and LL, respectively. The eclosion rhythm showed a tau of 23.33 +/- 0.63 h (mean +/- 95% CI) in DD, and eclosion was not rhythmic in LL. The tau of the oviposition rhythm in DD was significantly greater than that of the eclosion rhythm. The eclosion rhythm of all 10 replicate vials entrained to the three periodic light regimes, T20, T24, and T28, whereas the oviposition rhythm of only about 24 and 41% of the individuals entrained to T20 and T24 regimes, respectively, while about 74% of the individuals assayed in T28 regimes showed entrainment. Our results thus clearly indicate that the tau and the limits of entrainment of eclosion rhythm are different from those of the oviposition rhythm, and hence this reinforces the view that separate oscillators may regulate these two rhythms in D. melanogaster.     

Daily growth increments in the otoliths of Atlantic salmon parr, Salmo salar L., and the influence of environmental factors on their periodicity

Daily increments were demonstrated in the sagitta otoliths of fast- and slow- growing Atlantic salmon parr, Salmo salar L., when held under natural photoperiod and temperature. Otolith increments continued to be deposited at a daily rate when fish were held under constant light and/or temperature and on single or multiple feeding regimes. However abnormally short photoperiods of 6L: 6D induced two increments per day. The results suggest that an endogenous rhythm, synchronized lo light/dark transitions within a 24 h period, controls otolith increment deposition.     

Circadian locomotory rhythm and the influence of moulting in Australian field cricket nymphs

ABSTRACT. Evidence is presented for a circadian control of locomotory activity in the larval stadia of the cricket, Teleogryllus commodus Walker. Under light—dark cycles (LD), maximal activity occurs around the L/D transition and/or in the hours preceding it. Free-running rhythm patterns longer than 24 h are observed in constant light. Re-entrainment to phase advances in the LD cycle is also accompanied by several transient cycles. However, free-running rhythms under constant darkness or transients when exposed to LD cycle delays were not found. LD cycles during the eighth stadium set the phase of a free-running rhythm in the adult, even if the nymph does not show a rhythm. Nymphal activity is often erratic and is disrupted periodically by the moulting cycle, but moulting does not interrupt the operation of the circadian system. The daily timing of the moult itself is not under circadian control.     

Comparison of rectal and vaginal temperature daily rhythm in dogs (Canis familiaris) under different photoperiod

To investigate the daily rhythm of rectal and vaginal temperature, we used six mongrel dogs with a mean body weight of 15±3 kg, aged between 2 and 3 years old. Rectal and vaginal temperatures were recorded every 3 h over 48-h period during three different lighting regimes: natural light/dark (L/D) cycle (sunrise 06:25, sunset 17:05), constant light (L/L) and constant darkness (D/D). A daily rhythm of rectal temperature was observed in both days of monitoring in all experimental conditions. Vaginal temperature showed a daily rhythmicity in L/D and D/D cycle. During the L/L cycle, daily rhythm was disrupted. Rhythmic parameters (MESOR, amplitude, acrophase and robustness) did not change between the different photoperiod and the site of temperature collection. In conclusion, the monitoring of vaginal temperature can be considered a valid alternative to the monitoring of body temperature as well as rectal temperature under natural lighting conditions in canine medicine.     

Temporal change of the aggregation response in Triatoma infestans

Adults and larvae of Triatoma infestans spend daylight hours assembled in shaded places. An assembling factor has been demonstrated in the excrement of this species. We analysed different aspects of the dynamics of the response of bugs. Recently fed insects do not aggregate around faeces. They start to show a significant assembling response from the 8th hour after feeding onwards. Just deposited faeces do not evoke assembling, but a significant rejection instead. This reaction switches 3 h after deposition, when the faeces become attractive to the insects. The attractiveness of faeces persists for about 10 days and can be recovered after this time by rehydration. These findings are discussed in relation to the biological role of faeces and the dynamics of the use of refuges by T. infestans.     

Turnover of rod photoreceptor outer segments. II. Membrane addition and loss in relationship to light

The rate of disk addition to rod outer segments (ROS) varies widely in Xenopus laevis tadpoles kept in cyclic light (12L:12D). When measured as radioactive band (3H-band) displacement during the 2nd day after injection of [3H]leucine, 75% of the daily increment of displacement occurred during the first 8 h of light. During the same interval, the number of open disks at the ROS base increased more than threefold. During the last 8 h of darkness, 3H-band displacement was undetectable and the number of open disks was reduced. These observations suggest the possibility that disk addition may occur discontinuously. During the 3rd and 4th days after injection of [3H]leucine, maximal displacement of the 3H-band occurred later in the day than on the 2nd day, its movement no longer corresponding to the increase in open disks. This delay in 3H-band displacement may reflect a time delay as a result of propagation of compressive stress in an elastic ROS system. Maximal disk loss from ROS as reflected in counts of phagosomes in the pigment epithelium occurred within 1 h of light exposure, and phagosome counts remained high for 4 h before declining to a low level in darkness. Modified lighting regimes affected the daily rhythms of shedding and disk addition differently, suggesting that control mechanisms for the two processes are not directly coupled. During 3 days in darkness, disk addition was reduced 50% compared to controls (12L:12D), whereas shedding was reduced by about 40%. Although reduced in level, shedding occurred as a free-running circadian rhythm. There was no evidence of rhythmicity of disk addition in darkness. In constant light, the rate of disk addition was not different from controls, but shedding was reduced by about 80% after the 1st day. This resulted in a 21% increase in ROS length. Among animals kept on a 2.5L:21.5D cycle, the rate of disk addition was reduced by 40% while shedding was maintained near control levels, resulting in a slight decrease in ROS length. These observations indicate that normal shedding requires alternating light and darkness, and that the daily rhythm of disk addition is due primarily to daily stimulation by light.     

Latitudinal variation in eclosion rhythm among strains of Drosophila ananassae.

Eclosion rhythm parameters of D. ananassae strains originating between 8 degrees-34 degrees N were highly variable and latitude dependent. In the field under naturally fluctuating light intensity, temperature and R.H., the amplitude of the rhythm was high and the eclosion gate was narrow; however, under the naturally fluctuating light intensity but at constant temperature and R.H., the amplitude of the rhythm was lowered and the width of eclosion gate was widened. The eclosion rhythm entrained to light-dark (LD) cycles ranging from LD 6:18 to LD 18:6, the width of the eclosion gate was decreased and increased in the short and long photoperiods respectively. Among the strains, both the phase angle difference (psi, the time from lights-off in a 24 hr LD cycle to the eclosion median) and the period of free-running rhythm (tau) in constant darkness varied by about 3 hr and the amplitude of the rhythmicity (Amp) by about 10%. Lower latitude was correlated with late psi (r = -0.69), long tau (r = -0.88) and high Amp value (r = -0.95).     

Entrainment of Circadian Wheel-Running Rhythms of the Northern Brown Bandicoot, Isoodon macrourus, by Daily Restricted Feeding Schedules

Northern brown bandicoots (Isoodon macrourus) were subjected to restricted feeding for 3 h in the middle of the light period of a 14: 10 light/dark cycle and immediately following this in constant dark. When feeding was restricted to the middle of the light period of the light/dark cycle, all bandicoots maintained a nocturnal activity rhythm. In addition to the nocturnal rhythm, a few bandicoots showed meal-anticipatory activity during the light period. In bandicoots that did not show meal-anticipatory activity, diurnal activity was sometimes evident either during or shortly after the daily meal time. The observation of meal-anticipatory activity in some bandicoots suggests that this species may have a mechanism separate from the light-entrainable mechanism that allows the daily anticipation of periodically available food sources. In the next stage of the experiment, which was in constant dark, the meal was presented at the same time of day as it had been in the previous stage. In all bandicoots, the previously light-entrained component of activity free-ran and was eventually affected by the restricted feeding schedule to some degree. Bandicoots showed weak entrainment and relative coordination, suggesting that restricted feeding is a weak zeitgeber in this species. Evidence also suggesting that two separate but coupled pacemakers control the activity rhythms of the bandicoot was that (a) bandicoots simultaneously showed free-running light-entrainable rhythms and meal-entrained anticipatory rhythms; (b) in several bandicoots, the light-entrainable rhythm was phase advanced when it free-ran through the meal time; and (c) in one bandicoot, meal-entrained anticipatory activity was forced away from the meal time when the previously light-entrained component of activity free-ran through it.     

Effects of feeding cycles on circadian rhythms in squirrel monkeys

Squirrel monkeys (Saimiri sciureus) were housed singly in cages equipped with a tree for climbing to measure locomotor activity, and with a movable food cup that could be arrested automatically. The animals were kept in continuous dim illumination (LL), twice interrupted by several weeks of entrainment by a light-dark (LD) 12:12 cycle. Apart from three control sections in which the food cups were unlocked continuously (ad libitum feeding), food was accessible for 3 hr per day only, with interfeeding intervals varying from 23 to 26 hr (periodic restricted feeding, or RF). During LD entrainment, the imposition of an RF schedule resulted in anticipatory behaviors, represented by increased tugs at the food cup and a pause in locomotor activity preceding the feeding time. In LL, the animals showed free-running circadian rhythms of locomotor and "feeding" activity that nearly always persisted when ad libitum feeding was replaced by RF. The period (tau) of the free-running rhythm was slightly modulated in relation to the varying interfeeding intervals (T), but entrainment was never achieved except in one test with an animal whose tau was very close to T. It is concluded that periodic availability of food represents an extremely weak zeitgeber, if any, for the circadian pacemaker of squirrel monkeys.     

The role of the ocelli in the phototactic behaviour of the haematophagous bug Triatoma infestans

In addition to compound eyes, most adult insects posses two or three simple eyes, the ocelli. The function of these photoreceptors remains elusive in most cases. Triatomine bugs posses two well-developed ocelli, located in a latero-dorsal position, behind the compound eyes. We tested the role of the ocelli in the phototactic behaviour of Triatoma infestans, by measuring the time spent by adult males in the dark half of an experimental arena, which had the other half illuminated. The occlusion of the ocelli or the compound eyes alone had little effect on the phototactic response of the bugs. Only those insects which had both their ocelli and compound eyes occluded showed a significant reduction in their negative response to light. The ability of the ocelli of T. infestans to mediate the phototactic response by themselves (i.e., not through the modulation of compound eyes sensitivity) constitutes the first report on this function in insects.     

A case for multiple oscillators controlling different circadian rhythms in Drosophila melanogaster

A population of the fruit fly Drosophila melanogaster was raised in periodic light/dark (LD) cycles of 12:12 h for about 35 generations. Eclosion, locomotor activity, and oviposition were found to be rhythmic in these flies, when assayed in constant laboratory conditions where the light intensity, temperature, humidity and other factors which could possibly act as time cue for these flies, were kept constant. These rhythms also entrained to a LD cycle of 12:12 h in the laboratory with each of them adopting a different temporal niche. The free-running periods (tau) of the eclosion, locomotor activity and oviposition rhythms were significantly different from each other. The peak of eclosion and the onset of locomotor activity occurred during the light phase of the LD cycle, whereas the peak of oviposition was found to occur during the dark phase of the LD cycle. Based on these results, we conclude that different circadian oscillators control the eclosion, locomotor activity and oviposition rhythms in the fruit fly D. melanogaster.     

CIRCADIAN ACTIVITY RHYTHMS AND SENSITIVITY TO NOISE IN THE MONGOLIAN GERBIL (MERIONES UNGUICULATUS)

Since consistent data on endogenous circadian rhythms of Mongolian gerbils are not available, the main aim of our study was to identify suitable conditions to receive stable and reproducible free-running rhythms of activity under different light intensities. Another objective was to determine the role of social cues as an exogenous zeitgeber in the absence of a light-dark (LD) cycle. We performed two long-term sets of experiments with adult male gerbils kept in climatic chambers under various photoperiods of at least 30 days each. In all cases, the time of lights on in the chambers differed from the daily starting hour of work in the animal house. Always, two animals per chamber were kept separately in cages with a running wheel while their activity was monitored continuously. During the first set, only three of eight animals developed intra- and interindividual variable free-running rhythms. The activity patterns seemed to be influenced by human activities outside, indicating high sensitivity to external factors. Subsequently, we damped the chambers and the room and restricted access to the room. In the following noise-reduced set, all gerbils developed comparable free-running rhythms of activity. We determined the mean of the free-running period τ, the activity-rest relationship α/θ and the amount of running wheel activity per day: τ = 23.7h ± 0.08h under low light (5 lux) and 25.5h ± 0.19h under high light intensities (450 lux); α/θ = 0.53 ± 0.08 under 5 lux and 0.34 ± 0.04 under 450 lux. The amount of daily activity was 12 times as high under 5 lux as under 450 lux. There was no indication that the two animals in one chamber socially synchronized each other. In conclusion, the pronounced rhythm changes in accordance with Aschoff's theory support the view that gerbils are mainly nocturnal animals. (Chronobiology International, 17(2), 137–145, 2000)