Adult Emergence Rhythm of Fruit Flies Drosophila melanogaster under Seminatural Conditions

In insects, the role of circadian clocks in the temporal regulation of adult emergence rhythm under natural conditions has not previously been reported. Here we present the results of a study aimed at examining the time course and waveform of emergence rhythm in the fruit fly Drosophila melanogaster under seminatural condition (SN). We studied this rhythm in wild-type and clock mutant flies under SN in parallel with laboratory condition (LAB) to examine (1) how the rhythm differs between SN and LAB, (2) what roles the circadian clock protein PERIOD and the circadian photoreceptor CRYPTOCHROME (CRY) play in the regulation of emergence rhythm under SN, and (3) whether there is seasonality in the rhythm. Under SN, wild-type flies displayed tightly gated emergence, peaking at "dawn" and gradually tapering down toward the evening, with little or no emergence by night, while in LAB, flies emerged throughout the light phase of light-dark (LD) cycles. The period loss-of-function mutant (per ( 0 )) flies were arrhythmic in LAB but displayed weak rhythmic emergence under SN. Under SN, cry mutants displayed less robust rhythm with wider gates, greater variance in peak timing, and enhanced nighttime emergence compared to controls. Furthermore, flies showed seasonal variation in emergence rhythm, coupled either to light or to humidity/temperature depending on the severity of environmental conditions. These results suggest that adult emergence rhythm of Drosophila is more robust in nature, is coupled to environmental cycles, and shows seasonal variations.     

Stability of Adult Emergence and Activity/Rest Rhythms in Fruit Flies Drosophila melanogaster under Semi-Natural Condition

Here we report the results of a study aimed at examining stability of adult emergence and activity/rest rhythms under semi-natural conditions (henceforth SN), in four large outbred fruit fly Drosophila melanogaster populations, selected for emergence in a narrow window of time under laboratory (henceforth LAB) light/dark (LD) cycles. When assessed under LAB, selected flies display enhanced stability in terms of higher amplitude, synchrony and accuracy in emergence and activity rhythms compared to controls. The present study was conducted to assess whether such differences in stability between selected and control populations, persist under SN where several gradually changing time-cues are present in their strongest form. The study revealed that under SN, emergence waveform of selected flies was modified, with even more enhanced peak and narrower gate-width compared to those observed in the LAB and compared to control populations in SN. Furthermore, flies from selected populations continued to exhibit enhanced synchrony and accuracy in their emergence and activity rhythms under SN compared to controls. Further analysis of zeitgeber effects revealed that enhanced stability in the rhythmicity of selected flies under SN was primarily due to increased sensitivity to light because emergence and activity rhythms of selected flies were as stable as controls under temperature cycles. These results thus suggest that stability of circadian rhythms in fruit flies D. melanogaster, which evolved as a consequence of selection for emergence in a narrow window of time under weak zeitgeber condition of LAB, persists robustly in the face of day-to-day variations in cycling environmental factors of nature.     

Simulating natural light and temperature cycles in the laboratory reveals differential effects on activity/rest rhythm of four Drosophilids

Recent studies under semi-natural conditions have revealed various unique features of activity/rest rhythms in Drosophilids that differ from those under standard laboratory conditions. An additional afternoon peak (A-peak) has been reported for Drosophila melanogaster and another species D. malerkotliana while D. ananassae exhibited mostly unimodal diurnal activity. To tease apart the role of light and temperature in mediating these species-specific behaviours of four Drosophilid species D. melanogaster, D. malerkotliana, D. ananassae, and Zaprionus indianus we simulated gradual natural light and/or temperature cycles conditions in laboratory. The pattern observed under semi-natural conditions could be reproduced in the laboratory for all the species under a variety of simulated conditions. D. melanogaster and D. malerkotliana showed similar patterns where as D. ananassae consistently exhibited predominant morning activity under almost all regimes. Z. indianus showed clearly rhythmic activity mostly when temperature cycles were provided. We find that gradually changing light intensities reaching a sufficiently high peak value can elicit A-peak in D. melanogaster, D. malerkotliana, and D. ananassae even at mild ambient temperature. Furthermore, we show that high mid-day temperature could induce A-peak in all species even under constant light conditions suggesting that this A-peak is likely to be a stress response.     

Seasonality of the endogenous activity rhythm in Talitrus saltator (Montagu) from a sandy beach in northeastern Tunisia

The endogenous activity of the sandhopper Talitrus saltator (Montagu) has been investigated in a population from a relatively exposed beach in northeast Tunisia. The animals were found to exhibit a nocturnal circadian rhythm of locomotor activity, with no evidence of a circatidal component. The rhythm was monitored in individual animals over the four seasons, under constant laboratory conditions and seasonal variation in free-running period; the signal-to-noise ratio and waveform of the rhythm have been investigated, as well as the incidence of rhythmic animals in the population. The variation in these rhythm parameters is considered in relation to changes in environmental conditions prevailing at the site of collection.     

Day length and evening temperature predict circannual variation in activity duration of the colony of the Indian cliff swallow,Hirundo fluvicola

In avian species, circannual rhythms, in a number of biological variables, including locomotor activity, have been studied under both laboratory and natural environmental conditions. However, information on circannual rhythm in daily activity duration of Indian cliff swallow (Hirundo fluvicola) is not available. It is a communal mud nesting, non-migratory species and inhabits low under arch bridges. Although it figures in the IUCN Red List under the least concerned category, it is important to study its behavioral ecology that may be of utmost importance from conservation stand point. In the present investigation, we examined the circannual rhythm in daily activity duration in this species at a communal roosting site under Kharoon river bridge on the Raipur–Bhilai highway (21°15′07.54′′N; 81°32′30.65′′E), Raipur, India, for eight consecutive days, every month from August 2009 to October 2011. On each study day, observations were made in two sessions, morning and evening. We monitored and recorded emergence time of the first bird from the nest in the morning and entry time of the last bird to the nest in the evening. We also recorded the ambient temperature and humidity of the study site simultaneously. Average daily activity duration (the time difference between the first bird’s emergence time and the last bird’s entry time from/to the nest, respectively) was computed for each month. A significant circannual rhythm in activity duration was validated. The peak of activity rhythm occurred on 22nd June with a spread between 19th June and 25th June. The activity duration was the shortest in winter and the longest in summer. The daily activity duration was positively correlated to the day length, sunset time, and morning and evening temperatures; whereas negatively correlated with the sunrise time, and morning and evening humidity. The multiple linear regression models suggest that day length alone explains 96% of the total variance in activity duration, whereas only 0.5% variance was attributable to evening temperature. We conclude that the day length is the strongest predictor of circannual variation in daily activity duration of the colony of Indian cliff swallow; whereas the evening temperature has very little effect. Further, we recommend that comparable studies under natural conditions might be very helpful to explore the effect of environmental cues on other intriguing behavioral decisions made by this and other avian species.     

The role of endogenous and exogenous factors in regulation of synchronous emergence of Trichogramma embryophagum Hartig and T. principium Sug. et Sor. (Hymenoptera, Trichogrammatidae)

The interaction of exogenous and endogenous factors affecting the daily rhythm of emergence of adult parasitoids Trichogramma embryophagum and T. princupium from laboratory strains was studied. Under a constant temperature of 20°C and different photoperiods (photophase of 12, 16, or 20 h), both species demonstrated a distinct rhythm; the majority of adults emerged during 4 first hours after light-on. The fraction of individuals emerged during this period depended on the photoperiod and was different in both the species studied. Experiments with non-periodic regimes (constant light or constant darkness, 20°C) demonstrated that the rhythm is based on the endogenous circadian oscillations that are synchronized with the external photoperiod. The time of emergence however, was not rigidly “programmed” and could be modified by the direct influence of the light and temperature Light-on and/or temperature increase (from 20° up to 30° C) induced the emergence of a certain percentage of individuals virtually at any time of the 24-h-long cycle. The magnitude of the response, estimated as the fraction of adults emerged during 2 h after the stimulation, changed daily under the control of the circadian oscillation. Thus, the plasticity of the daily dynamics of emergence was controlled (to a certain extent) by the endogenous rhythm of sensitivity to the stimuli. The interspecific differences in the relative sensitivity to light-on and temperature increase were revealed. When light and temperature stimuli were applied simultaneously, they caused a synergistic effect, exceeding a simple sum of separate impacts. These results demonstrate an important role of exogenous factors in control of the emergence rhythm in Trichogramma and can also be used in elaboration of methods of regulation and synchronization of emergence in these parasitoids.     

Intra- and inter-individual variability in the circadian rhythm of body temperature of rats,squirrels, dogs,and horses

In order to explore the often neglected issue of the relationship between intra- and inter-subject variability of circadian rhythms, we evaluated the variability in four parameters of the circadian rhythm of body temperature under controlled laboratory conditions. To avoid the bias of potential selection of an idiosyncratic species, we conducted the study on four different species: the laboratory rat, the thirteen-lined ground squirrel, the domestic dog, and the horse. In rats and squirrels, three of the four parameters of the body temperature rhythm (mean level, amplitude, and regularity of waveform) showed greater inter-subject variability than intra-subject variability. The two variabilities were not different from each other in dogs and horses. Intra- and inter-subject variabilities of acrophase (time of peak) were not significantly different in any of the species, and their magnitudes were similar in all species, which suggests that acrophase is a very dependable parameter in the analysis of circadian rhythms, even though its overall variability is not particularly low.     

温、湿度对南美斑潜蝇和美洲斑潜蝇羽化的影响

在实验条件下 ,研究了温湿度对南美斑潜蝇Liriomgzahuidobrensis和美洲斑潜蝇Liriomgzasativae羽化的影响。结果表明 ,温度对 2种斑潜蝇日羽化节律有显著的影响。随温度的升高 ,成虫的日羽化时段相应缩短 ,日羽化高峰期相应提前。且 2种斑潜蝇的羽化均集中在 1 2 :0 0之前。 2种斑潜蝇羽化所需要的最适温度 :南美斑潜蝇为 2 0℃ ,美洲斑潜蝇为 2 5℃ ;羽化所需要的最适相对湿度 :南美斑潜蝇为90 % ,美洲斑潜蝇为 1 0 0 %。     

Adult emergence rhythm of the egg-parasitoid Telenomus busseolae

The adult emergence rhythm of Telenomus busseolae, an egg parasitoid of Sesamia nonagrioides, was examined when parasitoids were exposed to different light-dark regimes. Most of the adult parasitoids emerged throughout the whole period of the photoperiodic cycle. Peak male emergence occurred 2–5 hours earlier than that of females. Adult emergence was asynchronous in continuous darkness or light. However, regimes of alternative light and dark phases such as L4:D20, L8:D16, L12:D12, L16:D8 and L20:D4 h generated a population rhythm with a period length of 24 hours. The peak of the emergence activity moves from the scotophase to the middle of the photophase with an increase of the photophase from 4 to 20 h. Rhythmical activity of adults was synchronised within 2 cycles when immature stages of parasitoid grow under continuous light conditions (LL) and then transferred to L12:D12. Moreover, emergence rhythm persisted and continued in a free-run with a period length of less than 24 hours by transferring a rhythmic culture from L12:D12 h to LL or RR (continuous red light) conditions, indicating the existence of a circadian rhythm. The ecological implications of the expression rhythm relate to better survival of the parasitoids.     

THE ANALYSIS OF THE DIVISION RATES OF CILIATES

1. Analysis of the division rates of Paramecium aurelia (mutant), Blepharisma undulans, and Histrio complanatus grown separately in pedigree isolation culture with the same culture medium, and in the same room at any given time, for a period of 3 years, discloses a secular trend and a seasonal rhythm for each organism. The seasonal rhythm is a yearly cycle with a maximum during July. 2. After removal of the effects of trend and seasonal rhythm, no correlation is found between the division rates of the several organisms. The distribution of the division rates is then one of chance order, except for large deviations known to be associated with changes in the culture technique. Each organism has a division rate varying independently of the others. 3. Consequently, seasonal rhythm alone has forced similar variations in the division rates of these three protozoans. The seasonal effect is gradually lost when the animals are raised for several years under laboratory conditions. Examination of the literature discloses other similar cases. 4. It is clear that unless all of the conditions of experiment are kept constant, one must analyze all protozoan division rate data in some such manner as that here presented before any conclusions may be drawn as to the existence of "cycles" or "rhythms."     

Management of pine wilt disease vectoring Monochamus alternatus adults using spray and soil application of Metarhizium anisopliae JEF isolates

Chemical control is widely used to control the Japanese pine sawyer beetle, Monochamus alternatus, but strong chemical regulations require an environmentally sound management strategy. In this work, we investigated the use of entomopathogenic fungi and their application as a means of practical pest management. Thirty-two diverse species of fungal isolates were assayed against adult pine sawyer beetles using a contact method under laboratory conditions, and four isolates showed over 70% virulence consequently. These isolates, two each of Beauveria bassiana and Metarhizium anisopliae were sprayed on the adult beetles at 1 × 10⁷ conidia/ml in plastic containers, respectively. The M. anisopliae-treated adult beetles showed 67% mortality. M. anisopliae isolates JEF-197 and JEF-279 demonstrated dosage-dependent insecticidal activity. Following the laboratory experiments, semi-field trials were conducted in young pine trees under high (RH 94%) and low (RH 35%) humidity conditions. In the high humidity conditions, most of the adult beetles stayed on the top of the branches. When the two M. anisopliae isolates were sprayed on the beetles, they showed ca. 50–70% insecticidal activity 11 days after application. In contrast, in low humidity conditions, the adult beetles tried to move off the branches and onto the soil. When the beetles reached the JEF-197 and JEF-279-treated soil, we measured >90% insecticidal activity. This work suggests that M. anisopliae was the most virulent entomopathogenic fungus against adult Japanese pine sawyer beetles, and this forest insect could be ecologically controlled by the spray and soil application of the M. anisopliae isolates.     

栗山天牛成虫羽化与温湿度的关系

栗山天牛Massicus raddei (Blessig)是危害我国东北辽东栎Quercus liaotungensis和蒙古栎Q.mongolicus天然林的重大害虫,生活周期长,3a发生1代,而且生活史整齐,在东北林区的辽宁、吉林和内蒙发育历期基本一致,因而绝大部分的成虫羽化时期出现在同一年份.为了有效防治栗山天牛,利用栗山天牛成虫具有趋光性这一特点,研究发明了诱集捕杀成虫的专用黑光灯,诱杀了大量的栗山天牛成虫,吉林省2008年共诱杀栗山天牛成虫达24.16t.根据每天晚上所诱到的栗山天牛的成虫数量(平均lkg栗山天牛成虫数量为430头,按此折合成重量kg统计),分析栗山天牛在成虫发生期其羽化数量与温湿度因子的关系.结果表明,栗山天牛成虫的羽化高峰期集中在7月下旬至8月上旬;成虫的羽化与林间气温呈显著的正相关关系,与相对湿度则呈显著负相关关系.研究对栗山天牛成虫的发生量与温湿度的关系进行了拟合.研究结果为我国东北林区栗山天牛成虫羽化年份预测预报成虫出现和发生期以及在成虫发生期开展防治提供了依据.     

Ontogeny of an endogenous rhythm in Eurydice pulchra

Field catches of Eurydice pulchra (Leach) from an Isle of Man beach indicated that adult animals were active in the water column solely during spring tide periods, whereas juvenile animals swam abundantly at high tide irrespective of lunar phase. Under constant conditions in the laboratory, animals kept in actographs with sand showed a circa-tidal rhythm of spontaneous emergence and swimming activity. In adults, the activity was greatest during “expected” night-time high-waters and was expressed solely after collection during spring tide periods. The rhythm was independent of sex, season or reproductive status. Juveniles showed no such diurnal modulation of the rhythm and the pattern was expressed throughout the semi-lunar cycle. These ontogenetic differences in behaviour are discussed in the light of previous work on the rhythm, and in terms of the differing habits of the mature and immature animals.     

Seasonal diapause development and diapause termination in the European skipper, Thymelicus lineola (Ochs.)

Eggs of the European skipper, Thymelicus lineola, containing pharate first-instar larvae were brought into the laboratory at the beginning of each month from September to May and maintained at 20°C, 14L:10D. No emergence was observed within 40 days of collection before December. From that time on, while total emergence was similar, the duration of the emergence period decreased over the season from > 120 days in December to < 14 days in May. There was however no marked change in the seasonal supercooling point values of diapausing larvae.The rate of emergence from eggs collected in March was influenced by photoperiod, being slower under short-day conditions. However for any given photoperiod tested, the rate of emergence increased with an increase in temperature. The possible adaptive significance of a two-phase diapause, where diapause termination is photoperiodically controlled in species that overwinter as an egg or as a pharate first-instar larva within the egg chorion in exposed habitats, is discussed.     

Photoperiodic regulation of the diapause of the progeny in <Emphasis Type="Italic">Trichogramma embryophagum</Emphasis> Htg. (Hymenoptera,Trichogrammatidae): Dynamics of sensitivity to photoperiod at the immature stages of maternal females

Photoperiodic regulation of T. embryophagum progeny prepupal diapause was investigated under laboratory conditions. Maternal females developed at 20°C, the position of the photosensitive period was detected by transferring immature stages between the diapause-inducing “short” day (L:D = 12:12) and the diapause-averting “long” day (L:D = 20:4). Progeny generation developed at L:D = 12:12 under the near-threshold temperature of 13°, 14°, or 15°C. Experiments showed that, at least under the used photo-thermal regimens, only the pupal stage was sensitive to the photoperiod influence. The highest photosensitivity was recorded during the last two days before the adult emergence. In this critical period, even a single short day cycle induced diapause in most of individuals. However, the reaction to the long day was weaker: at least two long light days immediately preceding the maternal adult emergence were necessary to avert the diapause. If a short day was preceded by a long day by more than 2–3 days before the adult emergence, the stepwise photoperiodic response caused an increase in the tendency to diapause. The results of this study could be used in pest biocontrol practice for elaboration of optimal methods for Trichogramma mass rearing and storage and also for prediction of its seasonal cycles under natural conditions.     

Effect of thermoperiod amplitude on the adult eclosion rhythm of Indian meal moth,Plodia interpunctella (Lepidoptera: Pyralidae)

The thermoperiodic time cue is known to be one of several zeitgebers (Zt) that entrain an organism's biological rhythms. This study investigated the adult eclosion rhythm of the Indian meal moth, Plodia interpunctella, (Lepidoptera: Pyralidae) under various thermocycles in constant darkness. The insects were exposed to different thermoperiods (e.g. 25.5°C/24.5°C). Thermophase : cryophase ratios (TC) ranged from 4 h : 20 h to 20 h : 4 h. The peak in adult eclosion rhythm occurred at Zt 18.8 ± 4.4 (mean ± SD) on a given day and at Zt 0.7 ± 3.8 on the subsequent day under TC 4 h : 20 h and TC 20 h : 4 h conditions, respectively (start of temperature increase = Zt 0). Thus, longer thermophase periods delayed the peak in eclosion. The regression line obtained was then compared with that obtained under the temperature cycle of 30°C/20°C, and the two lines were found to lie parallel to each other. The peak in adult eclosion rhythm occurred earlier under the 25.5°C/24.5°C conditions than under the 30°C/20°C conditions. The results strongly suggest that the amplitude of the temperature cycle is an important factor determining the temporal position of the peak in adult eclosion. This is a significant finding among insect species.     

Circadian clock of Drosophila montana is adapted to high variation in summer day lengths and temperatures prevailing at high latitudes

Photoperiodic regulation of the circadian rhythms in insect locomotor activity has been studied in several species, but seasonal entrainment of these rhythms is still poorly understood. We have traced the entrainment of activity rhythm of northern Drosophila montana flies in a climate chamber mimicking the photoperiods and day and night temperatures that the flies encounter in northern Finland during the summer. The experiment was started by transferring freshly emerged females into the chamber in early and late summer conditions to obtain both non-diapausing and diapausing females for the studies. The locomotor activity of the females and daily changes in the expression levels of two core circadian clock genes, timeless and period, in their heads were measured at different times of summer. The study revealed several features in fly rhythmicity that are likely to help the flies to cope with high variation in the day length and temperature typical to northern summers. First, both the non-diapausing and the diapausing females showed evening activity, which decreased towards the short day length as observed in the autumn in nature. Second, timeless and period genes showed concordant daily oscillations and seasonal shifts in their expression level in both types of females. Contrary to Drosophila melanogaster, oscillation profiles of these genes were similar to each other in all conditions, including the extremely long days in early summer and the cool temperatures in late summer, and their peak expression levels were not locked to lights-off transition in any photoperiod. Third, the diapausing females were less active than the non-diapausing ones, in spite of their younger age. Overall, the study showed that D. montana clock functions well under long day conditions, and that both the photoperiod and the daily temperature cycles are important zeitgebers for seasonal changes in the circadian rhythm of this species.     

Comparison of Acyrthosiphon pisum (Homoptera: Aphididae) and Frankliniella occidentalis (Thysanoptera: Thripidae) as prey for the development and survival of Chrysopa pallens (Neuroptera: Chrysopidae)

The effects of two different prey species on the pre-imaginal development and survival of the green lacewing, Chrysopa pallens were determined under laboratory conditions. The developmental duration from first instar to adult emergence was shorter when larvae were fed on Acyrthosiphon pisum (22.56 ± 0.39 d) than on Frankliniella occidentalis (29 ± 0.45 d). The survival rate of lacewings from first instar to adult emergence was 22.6% higher when they were fed on A. pisum than when fed on F. occidentalis. The possible implications of these findings are applicable to mass-rearing of C. pallens and for understanding predator's biology and survival capacity in the presence of prey species evaluated and could be used in the development of an effective biological control program.