The thesis deals with characterisation and changes of energetic status in the spruce bark beetle, Ips typographus, during pre-diapause, diapause and post-diapause quiescence.

A diapausa comportamental em carrapatos é caracterizada pela perda temporária da agressividade do indivíduo, tendo como conseqüência o prolongamento do período sem alimentação, sendo detectada em larvas de Amblyomma cajennense. Esta espécie destaca-se como principal vetor da Febre Maculosa no Sudeste brasileiro. Este estudo objetivou avaliar os efeitos do fotoperíodo e da temperatura na regulação da diapausa comportamental em larvas não-alimentadas de A. cajennense. Para isso, fêmeas ingurgitadas foram obtidas de eqüinos naturalmente infestados de Janeiro à Fevereiro de 2005 e 2006. No laboratório, cada grupo experimental foi composto por oito fêmeas ingurgitadas colocadas em vaso contendo capim braquiária (Brachiaria decumbens). Cada vaso foi exposto a uma condição de temperatura e fotoperíodo dentro de incubadoras B.O.D. (Marconi MA 402). A fotofase foi estabelecida por 4 lâmpadas (Philips TLT 75RS Extra luz do dia 20w) e a escotofase pela ausência total de luz. Os parâmetros biológicos observados foram período de pré-postura, incubação dos ovos, de permanência das larvas debaixo do capim e de presença de larvas na ponta do capim (comportamento de busca pelo hospedeiro). Paralelamente, verificou-se o poder infestante de larvas em diapausa para frangos (Gallus gallus), em comparação com larvas em não diapausa. Os resultados obtidos demonstram que, sob variação de fotoperíodo com temperatura constante de 25oC, a duração do intervalo de pré-postura e de incubação dos ovos foi semelhante em todos os grupos experimentais (1 a 2 semanas para período de pré-postura e 5 semanas para período de incubação dos ovos). Observou-se que a combinação de fotoperíodo 14:10 (claro:escuro) induziu a diapausa, sendo que o fotoperíodo de 12:12 ou 10:14 determinaram o término da diapausa. Em outras combinações de fotoperíodo, onde larvas estiveram no fotoperíodo de 12:12 ou 10:14 desde a eclosão, a aglomeração nas pontas do capim ocorreu dentro das primeiras semanas de vida das larvas. Larvas mantidas em fotoperíodo 12:12 e 10:14 não entraram em dormência e larvas mantidas em condição de fotoperíodo de 14:10 permaneceram no solo por tempo maior que as outras. No entanto, a diminuição da temperatura de 25 para 15oC, no fotoperíodo constante de 14:10, induziu o término da diapausa. Em todos os experimentos realizados sob fotoperíodo de 10:14, nenhum dos regimes de temperatura foi eficiente para induzir a diapausa. Houve diferença no intervalo de incubação dos ovos entre grupos experimentais com temperatura de 25 e 20oC. Não houve diferenças significantes (P > 0,05) entre proporções de larvas em diapausa e em não diapausa que se ingurgitaram nos frangos, embora larvas em diapausa apresentaram período parasitário significativamente mais longo (P < 0,05). Conclui-se que a diapausa em larvas de A. cajennense é induzida somente se a condição de fotoperíodo estiver com maior número de horas de claro (14:10) durante a eclosão das larvas. Temperaturas de 20 ou 25oC não são capazes de induzir a diapausa, quando mantidas em fotoperíodo 12:12 ou 10:14. O término da diapausa é desencadeado tanto pela mudança de fotoperíodo (14:10 para 12:12 ou 10:14) como pela diminuição da temperatura de 25 para 15oC. / Behavioral diapause in ticks is characterized by the temporary loss of aggressiveness, and consequently, to the extension of the starvation period, as recently detected in Amblyomma cajennense larvae. This species is the main vector of spotted fever in southeastern Brazil. The present study aimed to evaluate the effect of photoperiod and temperature on the regulation of behavioral diapause of A. cajennense unfed larvae. For this purpose, engorged females were obtained from horses naturally infested from January to February in each of the years 2005 and 2006. In the laboratory, each experimental group was composed by eight engorged females, which were placed in a grass plot composed by the grass Brachiaria decumbens, held in a small basket (40 cm diameter). Each plot was exposed to a particular condition of temperature and photoperiod within a BOD incubator (Marconi MA 402). Photoperiod was provided by 4 lamps (Philips TLT 75RS 20w) and scotophase by total darkness. Evaluated tick biological parameters were preoviposition and incubation periods, as well as the period of time the larvae remained under the grass after hatching, and the period of time that larvae were seen questing on the top of the grass leaves (host questing). The infestation capacity of diapause larvae to feed on chickens (Gallus gallus) was compared to larvae without being in diapause. The results showed that under variation of photoperiod and constant temperature of 25oC, the duration of preoviposition and incubation periods were similar in all the experimental groups (1 to 2 weeks to preoviposition and 5 weeks for egg incubation). Photoperiod 14:10 (light: dark) induced recently hatched larvae to enter diapause, whereas photoperiod 12:12 or 10:14 determined the end of diapause. In other combinations of photoperiod, where larvae were in photoperiod 12:12 or 10:14 from the eclosion, formation of larval clusters on the top of grass leaves occurred within the first weeks after larval eclosion. Larvae kept in photoperiod 12:12 and 10:14 did not enter in dormancy, whereas larvae maintained in photoperiod 14:10 since eclosion remained in the soil under the grass for longer periods. However, the decrease in temperature from 25 to 15oC under constant photoperiod of 14:10 triggered the end of diapause. In all experiments performed under photoperiod of 10:14, none of the temperature regimes was effective to induce diapause. There were different incubation periods between experimental groups held at 25 and 20oC. There were no significant differences (P > 0.05) between the proportions of diapause and non-diapause larvae that engorged on chickens, although the period of parasitism of larvae in diapause was significantly longer (P < 0.05). As conclusions for the present study, diapause in A. cajennense larvae is induced only if the condition of photoperiod is with more hours of light (14:10) during larval eclosion. Temperatures of 20 or 25oC are not able to induce diapause, when kept in photoperiod 12:12 or 10:14. The end of diapause is triggered both by the change in photoperiod (14:10 to 12:12 or 10:14) and the decrease in temperature from 25 to 15oC.

Diapause is often an adaptation for survival during periods of harsh environmental conditions. Some diapausing stages do not terminate the dormancy once the favourable conditions are restored. Such prolonged diapause may be enforced by environment if a diapausing stage cannot be reached by the cues inducing termination of dormancy. However, it may also be an advantageous bet-hedging strategy to allow only a fraction of dormant stages produced in any given season to hatch the next time conditions become favourable. I tested whether such strategy can be observed in hatching patterns of dormant eggs of Daphnia obtusa - a cladoceran occurring in small Central European temporary waters. I investigated the influence of intensity of illumination on hatching success, and effect of isolating the eggs encased in ephippia from the sediment. Fraction of eggs terminating diapause, fraction of embryos successfully leaving the egg membranes, and timing of the response were assessed at 15 ˚C under four intensities of illumination (100% = 35µmol.m2 .s-1 , 75%, 50%, 25%; photoperiod 12h light: 12h dark) and in complete darkness for 21 days. My results support previous suggestions that there is no genetically-fixed bet-hedging strategy in D. obtusa. I observed high proportion of eggs which terminated diapause in all...

A diapause upregulated cDNA clone was isolated from a cDNA library generated from brain mRNA of diapausing Sarcophaga crassipalpis pupae. The clone hybridized to a 1600 bp transcript on a northern blot. The insert is 823 bp in length, has a tentative open reading frame of 615 bp, and codes for a 23 kDa protein. The clone has a high level of identity at the amino acid level with the four small heat shock proteins of Drosophila melanogaster. Northern analysis revealed no detectable expression of the transcript in diapause- or nondiapause-programmed wandering larvae, and only trace expression in nondiapausing pupae. But, the transcript was highly expressed beginning at the onset of diapause and continuing throughout diapause. Expression promptly decreased when diapause was terminated. In nondiapausing individuals the transcript was highly expressed in response to cold shock or heat shock, but temperature stress did not cause greater expression in diapausing pupae. The results imply that expression of this small heat shock protein, a response elicited by temperature stress in nondiapausing individuals, is a normal component of the diapause syndrome. The upregulation of this gene during diapause suggests that it plays an essential role during this overwintering developmental arrest.

The spider mite predator Phytoseiulus persimilis Athias-Henriot and the thrips predator Amblyseius cucumeris (Oudemans) are both commercially mass-reared for use as biological control agents for greenhouse pests. The ability to stockpile these mites in cold storage would greatly facilitate economical mass-production and distribution. In addition, these two species provide an opportunity for comparative studies of cold hardiness because P. persimilis originated in subtropical Mediterranean type climates and is thought to be incapable of entering diapause whereas A. cucumeris is widespread in temperate zones where adult females enter a reproductive diapause for overwintering. Temperature/mortality curves confirmed that both species are freezing intolerant in the traditional sense that supercooling points (SCPs), at which freezing of body fluids occurs, represent absolute lower lethal temperatures. Both species were capable of moderate supercooling, into the range of-20°C to -30°C, and both showed a trend of increasing SCP temperatures during development from egg to adult that suggested an inverse relationship between supercooling capacity and body size within species. The only exception to this trend was adult female A. cucumeris, which may be significant because this is the only life stage that is capable of diapause or survival of temperate winters. On the other hand, diapause induction and low temperature acclimation had little or no effect on supercooling capacity, and survival of nonacclimated mites at subzero temperatures above their SCPs was limited to very short periods of exposure, suggesting that the SCP represents a physical property of the mites rather than an adaptation for survival of exposure to subzero temperatures. Cold-storage survival of both species was optimum at 7.5°C, was greatly enhanced when a source of moisture was provided, and was enhanced even further when food was provided even though the mites were held at temperatures below their theoretical temperature threshold for development. Under these conditions, survival of P. persimilis was 80% after six weeks whereas that of A. cucumeris was only 35% after the same period of time. Furthermore, longevity and fecundity of P. persimilis after eight weeks of cold storage were comparable to mites taken directly from rearing cultures whereas oviposition by A. cucumeris after six weeks of cold storage was low and irregular compared to previously reported values. Temperature data from areas where P. persimilis survives outdoors indicate that this species is capable of surviving prolonged exposure to temperatures below 10°C and occasional exposure to subzero temperatures, and apparently does so without diapausing. On the other hand, A. cucumeris may be capable of surviving prolonged exposures to subzero temperatures, but only as diapausing adult females. The results of the current studies are consistent with these ideas and further suggest that nondiapausing A. cucumeris are less cold hardy than P. persimilis. / Graduate

No description available.

Diapause is a programmed developmental arrest that allows insects to survive harsh environmental conditions. The diapause state has been linked to the insulin signaling pathway. Insulin signaling has been associated with many physiological processes including aging. It is a working hypothesis that the diapause and aging programs have a common set of gene expression pathways via insulin signaling. Analysis of a heterologous microarray indicated that two genes involved in the insulin pathway were down regulated during diapause. Both of these genes, Pi3K68d and Pde6, along with the Insulin receptor (InR) were targets for further investigation. Putative gene products have been isolated, cloned, and sequenced. RNA interference experimentation was conducted to characterize the role of the putative InR gene products obtained.

No description available.

No description available.

The brown marmorated stink bug (Halyomorpha halys) (Stål) is known for overwintering in human homes. Studies were conducted to understand the behavior of adult bugs in response to characteristics of potential overwintering structures including gap sizes, light, and presence of dead conspecifics. In a test where bugs were placed in increasingly tight boxes, most bugs settled in gaps 4.5 - 5.5 mm high. None settled in a space less than 3.5 mm high. In boxes with constant tightness, bugs tended to move to the back, and primarily settled along walls and in corners. In both box types, bugs tended to turn and face the cavity entrance during settling. In tests of responses of disturbed overwintering bugs to common household light bulbs over the course of a year, bugs were taken from shelters and exposed to lit bulbs. Bug responses were seasonal, with attraction to light bulbs in spring and summer, but little or no consistent response in fall or winter. The lack of response occurred more often at night than in afternoons. Because not all bugs survive the winter, corpses can accumulate. Single shelter-seeking H. halys were found not to respond to the presence of dead aggregations from the previous years unless touch was possible, in which case aggregations were joined. Dead aggregations from the same year had a repellent effect. Presence of a single dead bug from the same year did not provide any survivorship advantage to overwintering bugs. Results suggest possible improvements to trap shelters. / Master of Science in Life Sciences / The brown marmorated stink bug (Halyomorpha halys) is known for overwintering in human homes. This research investigated the responses of these bugs to some characteristics of overwintering structures, including crevice tightness preference, light sources, and dead bugs leftover from previous years. Tightness experiments indicated that bugs in crevices with hard walls prefer to settle in spaces between 4.5 and 5.5 mm high. Bugs also tended to stay on the floor, go back as far as they could, and face the entrance of the crevice. These findings will help improve overwintering box designs for collection and research, and will help pest control professionals focus their efforts. When bugs were disturbed and exposed to common household light bulbs, they tended to move toward the bulbs in late summer and in spring, but responded less in fall and winter, and responded more often in the afternoon than at night. Bugs were also exposed to other dead bugs in several ways. When lone bugs were exposed to groups of dead bugs from previous years, they went to them only if they could touch them. Single dead bugs had less of an impact. Exposure to groups of dead bugs from the same year drove live bugs away. Overwintering brown marmorated stink bugs do not appear to eat other bugs that have recently died. These results suggest several possible improvements to the designs of trap shelters used for research and control.