Effects of photoperiod and low temperature on diapause termination in the yellow-spotted longicorn beetle (Psacothea hilaris)

Abstract. .The effects of photoperiod and low temperature on diapause termination in the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae), were examined using a population from Ino, Japan. Diapausing insects obtained by rearing larvae under short daylength (12 or 13 h) at 25^oC were subjected to various treatments. When the photoperiod was changed at the same temperature, diapausing larvae showed a long-day response with a critical daylength between 13.5 and 14h. The diapause was terminated and consequently pupation occurred if the daylength was longer than 13.5 h. Chilling the diapausing larvae at 10^oC for 30 or more days also terminated diapause in most larvae irrespective of the photoperiods during and after chilling treatment. In contrast, the post-chilling photoperiod had a critical effect on development of diapausing larvae chilled for only 15 days.     

Photoperiodic and temperature effects on the intensity of larval diapause in Sesamia nonagrioides

Abstract. The intensity of larval diapause in Sesamia nonagrioides Lef (Lepidoptera: Noctuidae) was investigated under laboratory conditions. Newly hatched larvae were exposed to different stationary photoperiods (from LD 7 : 17 h to LD 14 : 10 h), at a constant temperature of 25 °C. Diapause incidence was higher when larvae were exposed to daylengths shorter than the critical value (LD 12 : 12 h), whereas the within‐treatment variation in the larval period appeared to be significantly correlated with the photoperiod applied. The incidences of diapause and the duration of larval development were also measured after exposing larvae to short photoperiods (LD 8 : 16 h, LD 10 : 14 h or LD 12 : 12 h) in combination with various temperatures (20, 22.5 or 25 °C). Although an increase in the incidence of diapause appeared with the lowering of the temperature, no statistical differences were observed in the time needed for pupation within the photoperiodic treatments at the temperatures of 20 and 22.5 °C. Furthermore, when diapausing larvae were transferred to the long photoperiod of LD 16 : 8 h, they immediately proceeded to pupation, regardless of the photoperiod or the temperature to which they had been previously exposed, indicating that there were no differences in the intensity of diapause. Photoperiodic changes from LD 10 : 14 h to LD 12 : 12 h or to LD 14 : 10 h at different larval ages reduced the intensity of diapause with (a) early age of transfer and (b) increase of daylength. By contrast, when larvae were transferred from the long photoperiod of LD 14 : 10 h to shorter, such as LD 10 : 14 h or LD 12 : 12 h, a small increase in the intensity of diapause with the shortening of the daylength was apparent. These results support the hypothesis that insects may compare the duration of the photoperiod and could classify them as either longer or shorter in relation to the critical value.     

Effects of photoperiod and temperature on the termination of diapause in the univoltine seed wasp Eurytoma plotnikovi

Abstract Mummified pistachios containing fully grown diapause larvae of Eurytoma plotnikovi Nikol'skaya (Hym., Eurytomidae) were collected in early August and late September in coastal northern Greece and subjected to various photoperiod and temperature treatments, then maintained at 19 or 26°C and a long-day (LD 16:8 h), a changing, or a short-day (LD 10:14 h) photoperiod until pupation. In larvae of early August (beginning of diapause) subjected for 20 weeks to 19°C under a long, a changing, or a short photophase, followed by 19°C and a long photophase, 50% of the larvae pupated after 24, 18 and 13 weeks respectively. After exposure for 20 or even 12 weeks to a short photophase and low temperatures (10 or 4°C), pupation occurred after only 7–8 weeks and was more synchronous. The ranges of temperature for diapause development and post-diapause morphogenesis overlap. After exposure for 12 weeks to short days and low temperature, larvae of late September pupated much sooner under long days than under short days and sooner at 26° than at 19°C. E.plotnikovi depends on both temperature and photoperiod for diapause development, low temperature having a strong favourable effect on the earlier part and long day on the later part of diapause. In a few larvae of another pistachio seed wasp, Megastigmus pistaciae Walker, after a long enough period of low temperatures, diapause was terminated normally at 26°C and long days, or at 19°C and long or short days.     

Photoperiodic induction of larval diapause and temperature-dependent termination in a wild multivoltine bruchid,Kytorhinus sharpianus

A wild bean weevil,Kytorhinus sharpianus Bridwell (Coleoptera: Bruchidae), has a multivoltine life cycle and enters a hibernal larval diapause at the fourth instar under a short daylength (Shimada & Ishihara, 1991). Here, we investigated their diapause incidence under different photoperiods at 24°C and 27°C. The critical photoperiods for diapause induction were 14.5 h at 24°C and 14 h at 27°C. The stages susceptible to diapause-inducing stimuli were estimated by transferring larvae of various instars from long days to short days and vice versa. Then we investigated the incidence of larval diapause. The sensitive stage was estimated to be from the third to early fourth instar. Though larval diapause, which was induced under a short daylength, was terminated only by increasing the daylength, the termination was more synchronized by an exposure to a low temperature followed by increasing temperature, irrespective of photoperiod.     

Control of diapause in codling moth larvae

Diapause in a New Zealand strain of codling moth (Cydia pomonella Linnaeus [Lepidoptera: Olethreutidae]) was induced in larvae by photoperiods of 15 h or less. Once diapause had been initiated, it could not be terminated by any combination of conditions tested for at least 20 days after cocooning. In diapausing larvae a low rate of pupation occurred at 25 °C under a long day (18 h) photoperiod. A high rate of pupation was achieved under a long day regime when larvae were decocooned, and provided with apple as nourishment. Diapause could be terminated predictably in 94–100% of larvae by 1) conditioning at 15 °C and constant darkness for periods of 40–100 days, then 2) chilling at 2±2 °C and constant darkness for 20–50 days followed by 3) any post-chill condition periods at 25 °C, 18 h photoperiod. Complete diapause termination was achieved when 100 days conditioning was followed by 30 days or 50 days post-chill period. Under these conditions, 76% termination occurred in the post-chill period after 10 days, and 93% after 25 days.To terminate diapause in codling moth larvae, we recommend that a 100 days conditioning followed by 30 days chilling and 50 days post chilling periods be used.     

Photoperiodic control of larval diapause in the yellow-spotted longicorn beetle, Psacothea hilaris: analysis by photoperiod manipulation

The photoperiodic control of diapause induction in the larvae of the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe), was investigated using a west Japan-type population collected from Ino, Kochi Prefecture, Japan. In this population, the larvae expressed a long-day photoperiodic response with a critical daylength between 13.5 and 14 h at 25 °C ; under a long daylength, the larvae pupated after the 4th or 5th instar, while the larvae entered diapause under a short daylength after 2.3 additional molts on average. When the photoperiod was changed from a short (L12:D12) to a long (L15:D9) daylength, pupation occurred in most of the individuals irrespective of the time of the change. When the photoperiod was changed from long to short at 1 or 2 weeks after hatching, all of the larvae entered diapause, whereas when the photoperiod was changed at 5 weeks after hatching or later, most of the larvae pupated. The 2 weeks exposures to a long daylength against a 'background' of a short daylength at various times revealed that the larvae of this insect are most sensitive to the photoperiod from 4 to 6 weeks after hatching.     

Interactive effects of photoperiod and temperature on diapause induction and termination in the yellow-spotted longicorn beetle, Psacothea hilaris

Abstract. The interactive effects of temperature (20 °C or 25 °C) and photoperiod (LD 12 : 12 h or LD 15 : 9 h) on diapause induction and termination are investigated in the west‐Japan type yellow‐spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae). Larval diapause of P. hilaris is induced under three diapause‐inducing conditions (20 °C–SD, 20 °C–LD and 25 °C–SD), and the diapause larvae are transferred to one of four conditions (20 °C–SD, 20 °C–LD, 25 °C–SD or 25 °C–LD) for observation of pupation, which indicates termination of diapause. The intensity of diapause induced under the three conditions increases in the order 20 °C–SD < 25 °C–SD < 20 °C–LD, when assessed by the time course of pupation after the transfer. On the other hand, the effectiveness of the temperature–photoperiod combinations to terminate diapause is in the order 25 °C–SD (ineffective) < < 20 °C–LD < 25 °C–LD < 20 °C–SD. Among the temperatures (5, 10, 15 and 20 °C) examined, 15 °C is the most effective in terminating diapause under the short day; diapause in most larvae appears to have been completed in 15 days.     

Photoperiodic control of larval development in the semivoltine cockroachPeriplaneta japonica (Blattidae: Dictyoptera)

Periplaneta japonica is semivoltine, entering early diapause in any (except the first) larval instar before the last, and late diapause in the last instar. Early diapause was induced under a short day of 13 h or less at 28°C, and under both short and long daylength (12–16 h) at 20°C. The shorter the daylength and the lower the temperature, the younger the instar was entering early diapause. Early diapause was terminated by a long day (16 h) or a high temperature (28°C), after which larvae grew faster in short days than in long days until the last instar, when they again entered diapause, always in short days and frequently in long days. This late diapause was terminated also by an increase in daylength and was always followed by adult emergence. In this case, 13 and 14 h daylengths after exposure to 12 h daylength were as effective as 16 h daylength. Ourdoor samples collected in late autumn, winter and early spring at Hirosaki (40.5°N) comprised two distinct size groups, corresponding with the early and late diapause instars. Based on these results, the seasonal development strategy and intriguing aspects of the photoperiodic response in this cockroach are discussed.     

Effect of daylength and temperature on the larval diapause of the sunflower moth, Homoeosoma electellum

The effect of daylength and temperature on the regulation of the larval diapause of a central Missouri population of the sunflower moth, Homoeosoma electellum, was examined. Fully grown fourth-instar larvae exhibit a facultative diapause. Measurements of the effect of photoperiod on diapause induction revealed critical photoperiods of about 13 h 30 min light/day at 20°C, and between 11 h 45 min and 12 h light/day at 23°C. Third and fourth-instar larvae were shown to be the main sensitive stages for diapause determination. Daylength was also shown to be an important regulator of the rate of diapause development. A short day of LD 10:14 h permitted only a low rate of diapause development, whereas long days of LD 14:10 h and LD 16:8 h accelerated diapause development at 25 and 30°C. When long days were alternated with short days at 30°C the accelerating effect of long days on diapause development was not found. Systematic transfers of chilled diapausing larvae revealed an accelerated diapause development in groups transferred from 10 to 30°C LD 10:14 h, but diapause development was not accelerated in groups transferred from 10 to 30°C LD 16:8 h.     

Diapause induction and termination: north-south strain differences in Ostrinia nubilalis

Diapause induction and termination responses of a northern strain (Minnesota [MN]) of Ostrinia nubilalis were compared with those of a southern strain (Georgia [GA]). A thermoperiod in constant light (12 hr at 25 degrees C alternating with 12 hr at 4 degrees C) failed to induce diapause in GA larvae, but approximately 50% diapause induction was observed in the MN population. Moreover, the 50% of MN larvae that continued their development (i.e., underwent pupation and adult development) did so at a slower rate, as measured by days to pupation, than GA larvae. In the laboratory, diapausing MN larvae responded more slowly to the optimal light-dark (LD) cycle for terminating diapause, LD 16:8, than did GA larvae. In the field MN populations are univoltine (i.e., are characterized by one generation per year). A delayed termination response in the spring, coupled with a longer critical daylength for diapause induction as daylength decreases during late summer (earlier diapause) restricts the time during which development can occur as contrasted with GA populations. In addition, it is postulated that these two phenomena, coupled with a possibly slower growth rate in the MN insects as revealed under laboratory conditions, may collectively represent the basis for univoltinism in the field.     

Juvenile hormone changes associated with diapause induction, maintenance, and termination in the beet webworm, Loxostege sticticalis (Lepidoptera: Pyralidae)

At 22°C and under a long-day photoperiod of L:D 16:8, all the last fifth instar Loxostege sticticalis larvae undergo prepupal stage and pupate without diapause. Under a short-day photoperiod of L:D 12:12, in contrast, they all enter diapause with approximately 36 days diapause maintenance and then terminate diapause spontaneously, although only 44% of the larvae terminated diapause successfully. Changes in hemolymph juvenile hormone (JH I) titers of diapause-destined larvae across diapause induction, maintenance and termination were examined using HPLC, and were compared with those of non-diapause-destined larvae from the fifth instar through pupation. JH I titer of the earliest fifth instar diapause-destined larvae remained at a high level with a peak of 220.4 ng/ml, though it decreased continuously to a minimum of 69.0 ng/ml on day 5 in the fifth instar when the larvae stopped feeding to enter diapause. During the diapause maintenance, JH I titer of the mature larvae increased significantly and maintained a high level until day 31 in prepupae. JH I titer declined and fluctuated at low level from 5 days before pupation. In contrast, JH I titer of both the fifth instar non-diapause-destined larvae and prepupae remained and fluctuated at low level consistently, as well as decreased before pupation. These results indicate that diapause induction and maintenance in this species might be a consequence of high JH, whereas diapause termination can be attributed to low JH titer, which was in agreement with the hormonal regulation observed in many other larval-diapausing insects.     

Larval diapause of Monochamus urussovi and photoperiodic effects on larval development in tree bolts

To determine the larval diapause and the effect of photoperiod on development in Monochamus urussovi (Coleoptera: Cerambycidae), larvae were reared on Abies sachalinensis and Picea jezoensis logs and bolts. Larvae stopped developing in the final instar at 25°C and 16L : 8D (16 h light and 8 h dark) whereas an exposure to 5°C in the dark (134 days) following acclimation at 12°C under natural daylength led to adult emergence. When larvae were reared under 8L : 16D or 16L : 8D at 25°C with an intervening period of chilling at 5°C in the dark (112 days), a photoperiod of 8L : 16D induced a shorter time required for adult emergence after being returned to 25°C, and smaller adult body size than 16L : 8D.     

Role of photoperiod and temperature in the induction of overwintering pupal diapause in the spotted tentiform leafminer,Phyllonorycter blancardella

The role of photoperiod and temperature in the induction of overwintering diapause inPhyllonorycter blancardella (F.) (Lepidoptera: Gracillariidae) was examined in the laboratory and field using leafminers from commercial apple orchards in Ontario, Canada.P. blancardella exhibited a long-day response to photoperiod: long daylengths resulted in uninterrupted development whereas short daylengths induced diapause. The estimated critical photoperiod for diapause induction was L14.25∶D9.75. The larvae of leafminers destined to enter diapause took ca. 3× longer to complete development than the larvae of non-diapausing leafminers. The development prolonging effect of photoperiod decreased with decreasing daylength. Temperature modified the diapause inducing effect of photoperiod. At L14.25∶D9.75, diapause incidence was similar at 15 and 20°C but was lower at 25°C. Photoperiod also altered the normal relationship between development rate and temperature. At L14.25∶D9.75, the duration of larval development of diapausing leafminers was similar at 15, 20 and 25°C. Temperature alone is unlikely to have a role in the induction of diapause because leafminers exposed to natural late summer and fall temperature regimes and L16∶D8 did not enter diapause.     

Effects of photoperiod and temperature on diapause induction and termination in the swallowtail,Sericinus montelus

Abstract Sericinus montelus overwinters as diapausing pupae. In the present study, the effects of photoperiod and temperature on diapause induction and termination of diapause are investigated. The results obtained demonstrate that high temperature can reverse the effect of short day‐lengths on diapause induction. Under an LD 12 : 12 h photoperiod, all pupae enter diapause at 15, 20 and 25 °C, whereas all pupae develop without diapause at 35 °C. No pupae enter diapause under an LD 14 : 10 h photoperiod when the temperature is above 20 °C. Photoperiodic response curves obtained at 25 and 30 °C indicate that S. montelus is a long‐day species and the critical day‐length is approximately 13 h at 25 °C. At 25 °C, the duration of diapause is shortest when the diapausing pupae are maintained under an LD 16 : 8 h photoperiod and increases under LD 14 : 10 h and LD 12 : 12 h photoperiods. Under an LD 16 : 8 h photoperiod, the duration of diapause is shortest when the diapausing pupae are maintained at 25 °C, followed by 20 and 30 °C, and then at 15 °C. These results suggest that a moderate temperature favours diapause development under a diapause‐averting photoperiod in this species. The duration of diapause induced by an LD 12 : 12 h photoperiod is significantly longer at 25 °C than those at 15, 20 and 30 °C, and is shortest at 15 °C. At 25 °C, the duration of diapause induced by LD 6 : 18, LD 12 : 12 and LD 13 : 11 h photoperiods is similar and longer than 90 days. Thus, the diapause‐inducing conditions may affect diapause intensity and a photoperiod close to the critical day‐length has significant influence on diapause intensity in S. montelus.     

Photoperiodism and control of summer diapause in the Mediterranean tiger moth Cymbalophora pudica

Photoperiodic responses to both constant and changing photoperiods were studied in the Mediterranean tiger moth Cymbalophora pudica. Embryos, larval instars and prepupae were all sensitive to photoperiod, and the responses of larvae and prepupae to changing photoperiods were similar. At 23+/-2 degrees C, constant 24-h photoperiods with short photophases (11, 12h) induced a long diapause (mean 88days) whereas long photophases (14, 16h) induced a short diapause (mean 52days). A change to a longer or shorter photophase during larval development or during diapause caused a significant extension (up to a maximum of 138days) or shortening (down to a minimum of 10days) of diapause, respectively, but only when at least one of the photophases was longer than 14h. Thus, shortening and prolongation of photophase had an opposite effect than constant short and long photophases, respectively. Changes within the range of photophases of 10-14h did not cause a significant change in diapause duration.Experimental results enabled us to outline the mechanisms regulating voltinism and the duration of summer diapause. For the monovoltine cycle, cold autumn/winter temperatures slow down larval development, and prepupal aestivation starts relatively late (March, April). Prepupae are then kept in diapause by the increasing daylength (>14h after late April). Pupation is synchronized by decreasing daylength after summer solstice, and imagoes emerge in September/October. For the bivoltine cycle, when the autumn/winter temperatures are relatively warm, a certain portion of the population (depending on the individual rate of growth) may be diverted to a bivoltine life-cycle. In such a case, larval development is fast and short enough to allow an early start of diapause (prior to or during February). The duration of such early diapause is not influenced by changes in daylength (<14h); pupation occurs very early (April/May), and spring generation imagoes fly and oviposit in May/June. Summer larvae and prepupae live under decreasing daylength (>14h), which shortens their diapause to 20-30days. Imagoes of the autumnal generation thus occur in September/October, together with the univoltine portion of the population.     

Diapause termination in the millet stem borer, Coniesta ignefusalis (Lepidoptera: pyralidae) in Ghana as affected by photoperiod and moisture

Studies were conducted in northern Ghana in 1997 and 1998 on the effects of contact moisture and photoperiod on the termination of larval diapause in the stem borer, Coniesta ignefusalis (Hampson). Results obtained over the two years consistently showed that on its own, photoperiod had no significant effect on diapause termination. In combination with contact moisture, however, a long day photoperiod of 13L:11D accelerated diapause termination and increased the cumulative percentage pupation. Contact with moisture was crucial for diapause termination but its influence was more pronounced in larvae collected from the field after March. Generally, access to moisture earlier than April resulted in very high larval mortality. These observations, together with the fact that some of the larvae (12%) denied access to water throughout the experimental period subsequently pupated, suggest that moisture may not be the sole factor triggering diapause termination in C. ignefusalis. The implications of these findings are discussed.     

Consequences of diapause on post‐diapause development,reproductive physiology and population growth of Chilo partellus (Swinhoe)

We investigate the effects of diapause on post‐diapause development, reproductive physiology and population growth of Chilo partellus (Swinhoe) (Crambidae: Lepidoptera). Aestivating and hibernating larvae of C. partellus are exposed to diapause terminating conditions (consisting of an LD 12 : 12 h photocycle at 27 ± 1 °C and 65 ± 5% relative humidity with a fresh diet) to terminate the diapause and observations are made on percentage pupation, pupal duration and weight, adult reproductive performance and population growth parameters. We find that the diapause in C. partellus significantly reduces the percentage pupation and weights of pupae, ultimately lowering the weight and reproductive performance of adults. Reduced weights of adult females are found to be directly associated with a lower number of egg cells in ovaries. Nevertheless, the reproductive performance of C. partellus males is also found to be greatly affected in the diapause (hibernation and aestivation) experiencing population in terms of the deposition of a lower number of spermatophores and eupyrene sperm in the reproductive tracts of females compared with the nondiapausing population. The results of the present study clearly indicate that a reduction in longevity, fecundity and egg viability, as well as a reduced rate of deposition of spermatophores and eupyrene sperm, in a diapause experiencing population of C. partellus ultimately leads to a reduction in population growth parameters, thus having implications for bio‐ecology and population dynamics under a changing climatic scenario.     

Responses to stepwise photoperiodic changes for the larval diapause of the Indian meal moth Plodia interpunctella

Abstract The Indian meal moth Plodia interpunctella Hübner (Lepidoptera: Pyralidae) diapauses as a last‐instar (fifth) larva. At 30 °C, no larvae enter diapause under any photoperiodic conditions; at 25 °C, the photoperiodic response curve is a long‐day type with a critical length of approximately 13 h light; at 20 °C, diapause is induced moderately even under long days (> 13 h). Cumulative effects of short days or long days on diapause induction are determined by alternate, stepwise and gradually changing regimes of photoperiod at 25 °C. When the larvae are repeatedly exposed to LD 16 : 8 h and LD 12 : 12 h photoperiods every other day, the incidence of diapause is 37%. When the larvae are placed under an LD 16 : 8 h photoperiod for 2 days and then under an LD 12 : 12 h photoperiod for 1 day, it is 38 %. Exposure to an LD 16 : 8 h photoperiod for 1 day and then to an LD 12 : 12 h photoperiod for 2 days induces only 15% diapause. This may indicate that the photoperiodic information is not accumulated in a simple fashion despite the generally accepted hypothesis (i.e. photoperiodic counter). Larvae exposed to an LD 16 : 8 h photoperiod for 5 days after oviposition express a very high incidence of diapause even under short days between an LD 2 : 22 h and LD 12 : 12 h photoperiod. After 10 days exposure to an LD 16 : 8 h photoperiod, however, the short day does not induce diapause strongly. On the other hand, an LD 12 : 12 h photoperiod in the early larval life is highly effective in the induction of diapause. A gradual increase or decrease of photoperiod (2 min day^?1) shows that the direction of photoperiodic change does not affect the diapause determination.     

Effect of temperature on the termination of diapause in the univoltine almond seed wasp Eurytoma amygdali

Diapausing larvae of Eurytoma amygdali Enderlein (Hymenoptera, Eurytomidae) were collected in early August and late September. They were subjected to various photoperiod and temperature regimens for up to 20 weeks, then kept at L16:D8 and 19 °C for another 14 to 26 weeks for diapause to be terminated and pupation to take place. Photoperiod did not affect diapause completion. It was confirmed that the two morphologically distinct diapause stages have different temperature requirements for their completion. The first diapause stage was completed synchronously at temperatures between 16 and 19 °C. A higher temperature of 26 °C delayed diapause development. The second stage required lower temperatures between 4 and 10 °C. Spontaneous termination of diapause was observed at constant 19 °C. When applied to the first diapause stage for 20 weeks, low temperatures made the larvae refractory to subsequent intermediate temperatures. The first stage was thus maintained until a higher temperature of 26 °C made the larvae regain their ability to respond to the intermediate temperatures and complete this stage. Larvae grown in Retsou almonds had a higher diapause intensity than larvae grown in Truoito almonds. The results suggest that, in nature, the high temperatures of late summer and early autumn are likely to maintain the first diapause stage. Subsequently, the less warm temperatures of autumn allow the completion of the first stage by late autumn, and the low temperatures of late autumn and of winter allow the completion of the second diapause stage by mid winter.     

Factors influencing the duration and termination of diapause in the Warehouse moth, Ephestia elutella

The influence of environmental factors on the duration of diapause was evaluated in larvae of Ephestia elutella (Hübner) reared in short photo-periods at 25C or below. Termination of diapause was hastened by long photoperiods, high temperatures, long periods at low temperature, or exposure to fumigants. Diapause terminated rapidly under long photoperiods at 30 or 25C, but not at 20C. The critical photoperiod for the termination of diapause was similar to that for induction, lying between 13 and 16 h at 25C. The longest duration of diapause occurred in constant darkness (DD) at 20C. However, batches of larvae reared at 20C in DD pupated a little sooner than batches reared under LD, if both were transferred at the start of diapause to warm, long-day conditions. Long exposure to low temperature reduced the number of long photoperiods necessary for the rapid termination of diapause at high temperature. Samples of larvae brought to the laboratory at monthly intervals from an unheated outbuilding in which they were overwintering, required an average of c. 200 days to pupate in DD at 25C when transferred in December, compared with only 32 days when transferred in February or March. By comparison, batches transferred to LD 16:8 at 25C required 39 days when transferred in December and 20–24 days in February and March. Holding at low temperature for long periods also encouraged synchronous emergence of the sexes. Duration of diapause was generally shorter in a laboratory stock than in a stock collected from the field.