Timed receptor tyrosine kinase signaling couples the central and a peripheral circadian clock in Drosophila.

Circadian clocks impose daily periodicities to behavior, physiology, and metabolism. This control is mediated by a central clock and by peripheral clocks, which are synchronized to provide the organism with a unified time through mechanisms that are not fully understood. Here, we characterized in Drosophila the cellular and molecular mechanisms involved in coupling the central clock and the peripheral clock located in the prothoracic gland (PG), which together control the circadian rhythm of emergence of adult flies. The time signal from central clock neurons is transmitted via small neuropeptide F (sNPF) to neurons that produce the neuropeptide Prothoracicotropic Hormone (PTTH), which is then translated into daily oscillations of Ca2+ concentration and PTTH levels. PTTH signaling is required at the end of metamorphosis and transmits time information to the PG through changes in the expression of the PTTH receptor tyrosine kinase (RTK), TORSO, and of ERK phosphorylation, a key component of PTTH transduction. In addition to PTTH, we demonstrate that signaling mediated by other RTKs contributes to the rhythmicity of emergence. Interestingly, the ligand to one of these receptors (Pvf2) plays an autocrine role in the PG, which may explain why both central brain and PG clocks are required for the circadian gating of emergence. Our findings show that the coupling between the central and the PG clock is unexpectedly complex and involves several RTKs that act in concert and could serve as a paradigm to understand how circadian clocks are coordinated.

Novel regulation pathway of eclosion hormones in Tribolium castaneum by distinct transcription factors through the initiation of 20-Hydroxyecdysone.

Eclosion hormone (EH) is not only a key trigger of insect ecdysis, but is also involved in the regulation of important physiological processes such as development, diapause, metamorphosis and reproduction. EH is an ideal target for RNAi treatment and prevention of the Tribolium castaneum. However, two EH genes in Tribolium castaneum demonstrate distinct replication and functional conversion relationships, and the mechanisms of transcriptional regulation of eclosion hormone remain largely unexplored and poorly understood. In this study, the activity of highly active promoter fragments and potential transcription factors of TcEH and TcEHL were identified using the Dual-Luciferase reporter system and TANSFAC. TcSlbo and TcCAD were revealed to be important transcription factors for TcEH and TcEHL, respectively. Knockdown of TcSlbo failed to slough off the old epidermis of Tribolium castaneum and prevented them from developing into adults. Furthermore, we demonstrated for the first time that 20-hydroxyecdysone (20E) affects the expression of TcEH and TcEHL by regulating the transcriptional activities of TcSlbo and TcCAD. This study provides new insights into the transcription regulation of TcEH and TcEHL, their roles in insect growth and development, and the involvement of 20E in eclosion regulation, offering potential molecular targets for future pest management strategies.

The circadian clock gates Drosophila adult emergence by controlling the timecourse of metamorphosis.

The daily rhythm of adult emergence of holometabolous insects is one of the first circadian rhythms to be studied. In these insects, the circadian clock imposes a daily pattern of emergence by allowing or stimulating eclosion during certain windows of time and inhibiting emergence during others, a process that has been described as "gating." Although the circadian rhythm of insect emergence provided many of the key concepts of chronobiology, little progress has been made in understanding the bases of the gating process itself, although the term "gating" suggests that it is separate from the developmental process of metamorphosis. Here, we follow the progression through the final stages of Drosophila adult development with single-animal resolution and show that the circadian clock imposes a daily rhythmicity to the pattern of emergence by controlling when the insect initiates the final steps of metamorphosis itself. Circadian rhythmicity of emergence depends on the coupling between the central clock located in the brain and a peripheral clock located in the prothoracic gland (PG), an endocrine gland whose only known function is the production of the molting hormone, ecdysone. Here, we show that the clock exerts its action by regulating not the levels of ecdysone but that of its actions mediated by the ecdysone receptor. Our findings may also provide insights for understanding the mechanisms by which the daily rhythms of glucocorticoids are produced in mammals, which result from the coupling between the central clock in the suprachiasmatic nucleus and a peripheral clock located in the suprarenal gland.

A trade-off between desiccation resistance and developmental humidity for pupation height in the North Indian seasonal population of Drosophilid-Zaprionus indianus.

Drosophila larvae and pupae are vulnerable to seasonal abiotic stressors such as humidity and temperature. In wild low-humidity habitats, desiccation stress can occur as Drosophila larvae forsake wet food in search of a drier pupation site. Henceforth, the hypothesis that developmental humidity impacts pupation height, affecting larval and pupae water balance and fitness-related traits, was examined. Accordingly, warm-adapted Drosophilid- Zaprionus indianus from two seasons were reared under season-specific simulated conditions, with significantly varying relative humidity (summer RH: 40%; rainy RH: 80%), but nearly identical temperatures. A trade-off between pupation height and developmental humidity was observed. Drier summer conditions lead to pupae wandering farther from drier glass surfaces, resulting in higher pupation height (17.3 cm) while rainy pupae prefer wet food, resulting in lower pupation height (7.12 cm). Additionally, density-dependent pupation height was developmental humidity-specific, with most rainy-season pupae pupated on wetter food, while dry summer pupae pupated on glass surfaces or cotton. Nevertheless, flies from far pupation exhibited greater desiccation resistance, fecundity, and copulation duration than those from near pupation. The cuticular lipid mass of larvae and pupae was higher during far-than-near pupation, indicating decreased water loss rates compared to near-pupation. Finally, pupae eclosion (%) was unaffected by greater humidity (85%) in either season. Still, it considerably decreased at lower humidity (RH: 0% and 38%) for rainy pupae, further supporting the selection of low-humidity desiccation resistance in pupae. In conclusion, low humidity is crucial for survival of pre-adult stages of Zaprionus indianus under desiccation stress and for preference of pupation site.

Behavioral events and functional analysis of bursicon signal during adult eclosion in the 28-spotted larger potato ladybird.

To accommodate growth, insects must periodically shed their exoskeletons. In Manduca sexta, Drosophila melanogaster and Tribolium castaneum, Bursicon (Burs)/ Partner of bursicon (Pburs)-LGR2 signal is an indispensable component for the proper execution of ecdysis behavior during adult eclosion. Nevertheless, the behavioral events and the roles of bursicon signaling in other insects deserve further exploration. In the current paper, we found that the pupal-adult ecdysis in Henosepilachna vigintioctomaculata could be divided into three distinct stages, preecdysis, ecdysis and postecdysis. Preecdysis behavioral sequences included abdomen twitches, dorsal-ventral contractions and air filling that function to loosen the old cuticle. Ecdysis events began with anterior-posterior contractions that gradually split the old integument along the dorsal body midline, followed by freeing of legs and mouthparts, and culminated in detachment from pupal cuticle. Postecdysis behavioral processes contained three actions: perch selection and stretching of elytra and hindwings. RNA interference for HvBurs, HvPburs or Hvrk (encoding LGR2) strongly impaired wing expansion actions, and slightly influenced preecdysis and ecdysis behaviors. The RNAi beetles failed to extend their elytra and hindwings. In addition, injected with dsrk also caused kinked femurs and tibia. Our findings establish that bursicon pathway is involved in regulation of adult eclosion behavior, especially wing expansion motor programs. Given that wings facilitate food foraging, courtship, predator avoidance, dispersal and migration, our results provide a potential target for controlling H. vigintioctomaculata.

Divergent effects of climate change on the egg-laying opportunity of species in cold and warm regions.

Climate warming can substantially impact embryonic development and juvenile growth in oviparous species. Estimating the overall impacts of climate warming on oviparous reproduction is difficult because egg-laying events happen throughout the reproductive season. Successful egg laying requires the completion of embryonic development as well as hatching timing conducive to offspring survival and energy accumulation. We propose a new metric-egg-laying opportunity (EO)-to estimate the annual hours during which a clutch of freshly laid eggs yields surviving offspring that store sufficient energy for overwintering. We estimated the EO within the distribution of a model species, Sceloporus undulatus, under recent climate condition and a climate-warming scenario by combining microclimate data, developmental functions, and biophysical models. We predicted that EO will decline as the climate warms at 74.8% of 11,407 sites. Decreasing hatching success and offspring energy accounted for more lost EO hours (72.6% and 72.9%) than the occurrence of offspring heat stress (59.9%). Nesting deeper (at a depth of 12 cm) may be a more effective behavioral adjustment for retaining EO than using shadier (50% shade) nests because the former fully mitigated the decline of EO under the considered warming scenario at more sites (66.1%) than the latter (28.3%). We advocate for the use of EO in predicting the impacts of climate warming on oviparous animals because it encapsulates the integrative impacts of climate warming on all stages of reproductive life history.

Efectos divergentes del cambio climático sobre la oportunidad de desove de las especies en regiones cálidas y frías Resumen El calentamiento global puede tener un impacto considerable sobre el desarrollo embrionario y el crecimiento juvenil de las especies ovíparas. Es complicado estimar el impacto general que tiene el calentamiento global sobre la reproducción ovípara ya que los eventos de desove suceden durante la época reproductiva. El desove exitoso requiere que se complete el desarrollo embrionario y que el momento de eclosión sea favorable para la supervivencia de las crías y la acumulación de energía. Proponemos una nueva medida-oportunidad de desove (OD)-para estimar las horas anuales durante las cuales una puesta de huevos recién desovados produce crías que sobreviven y almacenan suficiente energía para invernar. Estimamos la OD dentro de un modelo de distribución de la especie Sceloporus undulatus bajo las recientes condiciones climáticas y bajo un escenario de calentamiento global mediante la combinación de datos microclimáticos, funciones del desarrollo y modelos biofísicos. Pronosticamos que la OD declinará conforme la temperatura aumente en 74.8% de los 11407 sitios. La disminución del éxito de eclosión y de la energía de las crías explicó más horas perdidas de OD (72.6% y 72.9%) que la presencia de estrés por calor en las crías (59.9%). Una anidación más profunda (a una profundidad de 12 cm) puede ser un ajuste conductual más efectivo para la retención de la OD que los nidos con mayor sombreado (50% de sombra) porque el primero mitigó por completo la declinación de la OD bajo el escenario de calentamiento en más sitios (66.1%) que el segundo ajuste (28.3%). Defendemos el uso de la OD en el pronóstico del impacto del calentamiento global sobre los animales ovíparos porque encapsula los impactos integrales que tiene el calentamiento global sobre todas las etapas de la vida reproductiva. 气候变化在寒冷和温暖地区对物种产卵机会造成不同影响.

Exposure to elevated temperature during development affects eclosion and morphology in the temperate Pieris napi butterfly (Lepidoptera: Pieridae).

Global warming has been identified as one of the main drivers of population decline in insect pollinators. One aspect of the insect life cycle that would be particularly sensitive to elevated temperatures is the developmental transition from larva to adult. Temperature-induced modifications to the development of body parts and sensory organs likely have functional consequences for adult behaviour. To date, we have little knowledge about the effect of sub-optimal temperature on the development and functional morphology of different body parts, particularly sensory organs, in ectothermic solitary pollinators such as butterflies. To address this knowledge gap, we exposed the pupae of the butterfly Pieris napi to either 23 °C or 32 °C and measured the subsequent effects on eclosion, body size and the development of the wings, proboscis, eyes and antennae. In comparison to individuals that developed at 23 °C, we found that exposure to 32 °C during the pupal stage increased mortality and decreased time to eclose. Furthermore, both female and male butterflies that developed at 32 °C were smaller and had shorter proboscides, while males had shorter antennae. In contrast, we found no significant effect of rearing temperature on wing and eye size or wing deformity. Our findings suggest that increasing global temperatures and its corresponding co-stressors, such as humidity, will impact the survival of butterflies by impairing eclosion and the proper development of body and sensory organs.

Unripe fruits of Litchi chinensis (Gaertn.) Sonn.: An overview of its toxicity.

Litchi (Litchi chinensis) is a widely consumed fruit that has been used in many food and health-promoting products worldwide. Litchi is a good source of nutrients including vitamin and minerals, dietary fibers, proteins, and carbohydrates. Of note, several studies have reported that the constituents of litchi fruits elicit antioxidant properties and help to maintain blood pressure, and reduce the risk of stroke and heart attack. An unclearly explained outbreak occurred in June 2019 in Muzaffarpur (Bihar), India resulted in the death of more than 150 children in a week, followed by a total of 872 cases and 176 deaths. This outbreak was associated with the consumption of Litchi fruits and the occurrence of acute encephalitis syndrome. In this high Litchi production region, a huge number of acute encephalitis syndrome cases have been registered in children in the past two decades with high mortality due to these neurological disorders linked to the consumption of litchi. While finding out the causes for this recurrent outbreak, whether or not it is caused by a virus or the phytotoxins of litchi is to be considered critical. Amongst the probable causes were observed to be methylene cyclopropyl acetic acid and hypoglycin-A found in unripe Litchi fruits which can cause hypoglycemia and as a plausible cause of AES outbreaks. This review addresses this recurrent outbreak in-depth exploring the possible causes and discusses the possible mechanisms by which phytotoxins of litchi such as hypoglycin A and methylene cyclopropylglycine which may elicit such toxic effects.

The role of neuropeptides in regulating ecdysis and reproduction in the hemimetabolous insect Rhodnius prolixus.

In ecdysozoan animals, moulting entails the production of a new exoskeleton and shedding of the old one during ecdysis. It is induced by a pulse of ecdysone that regulates the expression of different hormonal receptors and activates a peptide-mediated signalling cascade. In Holometabola, the peptidergic cascade regulating ecdysis has been well described. However, very little functional information regarding the neuroendocrine regulation of ecdysis is available for Hemimetabola, which display an incomplete metamorphosis. We use Rhodnius prolixus as a convenient experimental model to test two hypotheses: (1) the role of neuropeptides that regulate ecdysis in Holometabola is conserved in hemimetabolous insects; and (2) the neuropeptides regulating ecdysis play a role in the regulation of female reproduction during the adult stage. The RNA interference-mediated reduction of ecdysis triggering hormone (ETH) mRNA levels in fourth-instar nymphs resulted in lethality at the expected time of ecdysis. Unlike in holometabolous insects, knockdown of eth and orcokinin isoform A (oka) did not affect oviposition in adult females, pointing to a different endocrine regulation of ovary maturation. However, eth knockdown prevented egg hatching. The blockage of egg hatching appears to be a consequence of embryonic ecdysis failure. Most of the first-instar nymphs hatched from the eggs laid by females injected with dsRNA for eclosion hormone (dsEH), crustacean cardioactive peptide (dsCCAP) and dsOKA died at the expected time of ecdysis, indicating the crucial involvement of these genes in post-embryonic development. No phenotypes were observed upon corazonin (cz) knockdown in nymphs or adult females. The results are relevant for evolutionary entomology and could reveal targets for neuropeptide-based pest control tools.

Developmental Toxicology of Metal Mixtures in Drosophila: Unique Properties of Potency and Interactions of Mercury Isoforms.

Mercury ranks third on the U.S. Agency of Toxic Substances and Disease Registry priority list of hazardous substances, behind only arsenic and lead. We have undertaken uncovering the mechanisms underlying the developmental toxicity of methylmercury (MeHg), inorganic mercury (HgCl2), lead acetate (Pb), and sodium arsenite (As). To probe these differences, we used the Drosophila model, taking advantage of three developmental transitions-pupariation, metamorphosis, and eclosion-to differentiate potentially unique windows of toxicity. We elaborated dose response profiles for each individual metal administered in food and accounted for internal body burden, also extending analyses to evaluate combinatorial metal mixture effects. We observed all four metals producing larval lethality and delayed pupariation, with MeHg being most potent. Compared to other metals, MeHg's potency is caused by a higher body burden with respect to dose. MeHg uniquely caused dose-dependent failure in eclosion that was unexpectedly rescued by titrating in HgCl2. Our results highlight a unique developmental window and toxicokinetic properties where MeHg acts with specificity relative to HgCl2, Pb, and As. These findings will serve to refine future studies aimed at revealing tissue morphogenesis events and cell signaling pathways, potentially conserved in higher organisms, that selectively mediate MeHg toxicity and its antagonism by HgCl2.

Drosophila Choline transporter non-canonically regulates pupal eclosion and NMJ integrity through a neuronal subset of mushroom body.

Insect mushroom bodies (MB) have an ensemble of synaptic connections well-studied for their role in experience-dependent learning and several higher cognitive functions. MB requires neurotransmission for an efficient flow of information across synapses with different flexibility to meet the demand of the dynamically changing environment of an insect. Neurotransmitter transporters coordinate appropriate changes for an efficient neurotransmission at the synapse. Till date, there is no transporter reported for any of the previously known neurotransmitters in the intrinsic neurons of MB. In this study, we report a highly enriched expression of Choline Transporter (ChT) in Drosophila MB. We demonstrate that knockdown of ChT in a sub-type of MB neurons called α/ß core (α/ßc) and ϒ neurons leads to eclosion failure, peristaltic defect in larvae, and altered NMJ phenotype. These defects were neither observed on knockdown of proteins of the cholinergic locus in α/ßc and ϒ neurons nor by knockdown of ChT in cholinergic neurons. Thus, our study provides insights into non-canonical roles of ChT in MB.

RNA interference of eclosion hormone gene reveals its roles in the control of ecdysis behavior in Heortia vitessoides Moore.

Eclosion hormone (EH) is an important neuropeptide that regulates growth and development. This study predicted the EH gene (HvEH) of Heortia vitessoides Moore (Lepidoptera: Crambidae) from the transcriptome database and its expression patterns were determined using quantitative real-time polymerase chain reaction. HvEH was expressed in all developmental stages and especially in the head area. RNA interference-mediated silencing of HvEH (2 µg/individual) with double-stranded HvEH RNA (dsHvEH) was achieved within 48 hr. Abnormal phenotypes appeared in the pupa and adult stages. dsHvEH injection suppressed pupation and eclosion rates. HvEH expression increased upon treatment with 20-hydroxyecdysone but decreased at extreme temperatures. These results suggest that HvEH plays an essential role in ecdysis and wing formation in H. vitessoides.

Transcriptomic characterisation of neuropeptides and their putative cognate G protein-coupled receptors during late embryo and stage-1 juvenile development of the Aotearoa-New Zealand crayfish, Paranephrops zealandicus.

We de novo assembled a transcriptome for early life-stages of the Aotearoa-New Zealand crayfish, Paranephrops zealandicus, establishing the first genetic resource for this under-developed aquaculture species and for the Paranephrops genus. Mining of this transcriptome for neuropeptides and their putative cognate G protein-coupled receptors (GPCRs) yielded a comprehensive catalogue of neuropeptides, but few putative neuropeptide GPCRs. Of the neuropeptides commonly identified from decapod transcriptomes, only crustacean female sex hormone and insulin-like peptide were absent from our trinity de novo transcriptome assembly, and also RNA-sequence reads. We identified 63 putative neuropeptide precursors from 43 families, predicted to yield 122 active peptides. Transcripts encoding 26 putative neuropeptide GPCRs were identified but were often incomplete. Putative GPCRs for 15 of the neuropeptides identified here were absent from our transcriptome and RNAseq reads. These data highlight the diverse neuropeptide systems already present at the early development life stages sampled here for P. zealandicus.

Evaluation and comparison of the effects of three insect growth regulators on honey bee queen oviposition and egg eclosion.

Honey bees (Apis mellifera) are highly valued pollinators that help to ensure national food security in the United States, but reports of heavy annual losses to managed colonies have caused concerns and prompted investigations into the causes of colony losses. One factor that can negatively affect honey bee health and survival is agrochemical exposure. Investigations into the sublethal effects of agrochemicals on important metrics of colony health such as reproduction and queen fecundity has been limited by the availability of targeted methods to study honey bee queens. This work investigates the effects of three insect growth regulators (IGR), a class of agrochemicals known to target pathways involved in insect reproduction, on honey bee queen oviposition, egg hatching, and worker hypopharyngeal development in order to quantify their effects on the fecundity of mated queens. The reported results demonstrate that none of the IGRs affected oviposition, but all three affected egg eclosion. Worker bees consuming methoxyfenozide had significantly larger hypopharyngeal glands at two weeks of age than bees not fed this compound. The results suggest that although IGRs may not exhibit direct toxic effects on adult honey bees, they can affect larval eclosion from eggs and the physiology of workers, which may contribute to colony population declines over time.

Larval crowding results in hormesis-like effects on longevity in Drosophila: timing of eclosion as a model.

There is increasing evidence that stress during development can affect adult-life health status and longevity. In the present study, we examined life span (LS), fly weight, fecundity and expression levels of longevity-associated genes (Hsp70, InR, dSir2, dTOR and dFOXO) in adult Drosophila melanogaster flies reared in normal [low density (LD), ~ 300-400 eggs per jar] or crowded [high density (HD), more than 3000 eggs per jar] conditions by using the order (day) of emergence as an index of the developmental duration (HD1-5 groups). Developmental time showed a significant trend to increase while weight showed a significant trend to decrease with increasing the timing of emergence. In both males and females eclosed during first 2 days in HD conditions (HD1 and HD2 groups), both mean and maximum LSs were significantly increased in comparison to LD group. In males, mean LS was increased by 24.0% and 23.5% in HD1 and HD2 groups, respectively. In females, corresponding increments in mean LS were 23.8% (HD1 group) and 29.3% (HD2 group). In HD groups, a strong negative association with developmental time has been found for both male and female mean and male maximum LSs; no association with growth rate was observed for female maximum LS. The female reproductive activity (fecundity) tended to decrease with subsequent days of eclosion. In HD groups, the levels of expression of all studied longevity-associated genes tended to increase with the timing of eclosion in males; no differences were observed in females. On the basis of findings obtained, it can be assumed that the development in conditions of larval overpopulation (if not too extended) could trigger hormetic response thereby extending the longevity. Further studies are, however, needed to confirm this assumption.

The Drosophila apterous 56f mutation impairs circadian locomotor activity.

We investigated effects of apterous mutation ap56f on circadian locomotor activity, eclosion rhythms, and transcript levels of period and timeless in Drosophila. We investigated circadian locomotor activity and eclosion rhythms in ap 56fand wild-type flies, their F1 and F2 offspring, and wingless vestigial mutants and show that ap 56f disrupts circadian locomotor rhythms in a genetically recessive manner, that is not caused by the absence of wings. The ap blt strain also showed impaired circadian activity rhythms, providing independent evidence for a significant role of apterous in circadian locomotor rhythm expression. The ap 56f mutation did not disrupt a circadian eclosion rhythm or rhythmic expression of the period and timeless clock genes, indicating that apterous is not essential for circadian clock function, but is necessary for coupling locomotor activity to a circadian clock. Timeless transcription was reduced in ap 56f flies in 12:12 LD, suggesting that apterous may modulate core clock gene expression.

Comparative analysis of swallowtail transcriptomes suggests molecular determinants for speciation and adaptation.

Genetic determinants of speciation in closely related species are poorly understood. We sequenced and analyzed transcriptomes of swallowtail butterflies Heraclides cresphontes (northeastern species) and Heraclides rumiko (southwestern species), a pair of mostly allopatric sister species whose distribution ranges overlap narrowly in central Texas. We found that the two swallowtails confidently differ (FST > 0.5 for both species) in about 5% of genes, similarly to the divergence in another pair of swallowtail species Pterourus glaucus (southern species) and Pterourus canadensis (northern species). The same genes tend to diverge in both species pairs, suggesting similar speciation paths in Heraclides and Pterourus. The most significant differences for both species pairs were found in the circadian clock genes that were conserved within each species and diverged strongly between species (P-value < 0.01 and FST > 0.7). This divergence implied that adaptations to different climates and photoperiod at different latitudes or differences in mating behavior, including mating time and copulation duration, may be possible factors in ecological or behavioral-based speciation. Finally, we suggest several nuclear DNA regions that consistently and prominently differ between the sister swallowtail species as nuclear barcodes for swallowtail identification, with the best barcode being an exon from the protein TIMELESS.

Impact of short- and long-term heat stress on reproductive potential of Drosophila suzukii Matsumura (Diptera: Drosophilidae).

Drosophila suzukii Matsumura (Diptera: Drosophilidae) is an invasive vinegar fly of Asian origin now distributed throughout North America. Due to the unique morphology of females, this fly has become one of the most serious pests of thin-skinned fruits including blueberry, blackberry, cherry, raspberry, and strawberry. Prophylactic insecticide applications are commonly used to control this fly. A more sustainable approach to managing this invasive pest may not be possible without a clear understanding of the biology of this species under extreme environmental conditions. Specifically, high temperature is known to interfere with development and reproduction of drosophilids; however, the impact of high temperature on D. suzukii needs to be further investigated. The objective of the present study was to investigate the impact of exposure to constant and relatively short-term heat stress on reproductive success of D. suzukii, and potential for recovery. Results show that the development and reproduction of D. suzukii were negatively affected by constant and relatively short-term heat stress. Under constant heat stress, oviposition rate and adult lifespan decreased as temperature increased from 24 °C to 33 °C and reproduction was completely absent at 33 °C. Under relatively short-term heat stress, oviposition, pupation, and adult eclosion were significantly decreased as temperature increased from 28 °C to 34 °C. The short-term heat stress greatly reduced the fertility of both male and female D. suzukii which was recovered eight days after treatment. This study provides basic information on thermal biology of D. suzukii to help us better understand the trends commonly observed in D. suzukii trap captures in regions with hot summer conditions, and the results can be used in population models to predict its population dynamics in regions where high temperatures prevail during the field season.

An eclosion hormone-like gene participates in the molting process of Palaemonid shrimp Exopalaemon carinicauda.

Molting behavior is an important physiological process related to metamorphosis, growth, and reproduction in crustaceans. Previous studies indicated that the molting process was controlled by 20-hydroxyecdysone (20E) and upstream hormones, peptides, and environmental factors, which regulate 20E function. Eclosion hormone (EH) in insect is a kind of neuropeptide that is regulated by 20E and triggers ecdysis behavior at the end of molting process. However, the function of eclosion hormone gene during the molting process in crustaceans is still largely unknown. In the present study, an eclosion hormone-like gene EcEHL was identified from Exopalaemon carinicauda. The deduced amino acid sequence of EcEHL contained a signal peptide, a typical eclosion domain, and six conserved cysteine residues forming three putative disulfide bonds. EcEHL was predominantly expressed in the epidermis, gill, and eyestalk of shrimp. In situ hybridization analysis showed that EcEHL transcripts were localized in gill cells and in medulla externa X-organ, medulla terminalis X-organ, sinus gland, and lamina ganglionaris of eyestalks. During the molting process of shrimp, EcEHL showed the highest expression level in shrimp at the premolt stage. The expression level of EcEHL in shrimp at mid premolt stage was up-regulated by injection of exogenous 20E. Silencing of EcEHL using double-stranded RNA delayed both the molting process and ecdysis rate of E. carinicauda. Furthermore, injection of exogenous 20E to shrimp at mid premolt stage (D2) could remarkably speed up the molting process and also raise the ecdysis rate of E. carinicauda. The results revealed that EcEHL might participate in the molting process of shrimp and its expression was regulated by 20E. These data will help us to understand the molecular mechanism of molting in crustacean.

Genetic characterization of a Neisseria meningitidis cluster in Queensland, Australia.

Neisseria meningitidis serogroups B and C have been responsible for the majority of invasive meningococcal disease in Australia, with serogroup B strains causing an increasing proportion of cases in recent years. Serogroup Y has typically caused sporadic disease in Australia. In 2002, a cluster of 4 cases was reported from a rural region in Queensland. Three of these cases were serogroup C, with 1 case diagnosed by molecular detection only, and the fourth case was identified as a serogroup Y infection. Genomic analysis, including antigen finetyping, multilocus sequence typing (MLST), and core genome MLST, demonstrated that the serogroup Y case, though spatially and temporally linked to a serogroup C disease cluster, was not the product of a capsule switch and that one of the serogroup C isolates had a deletion of the entire porA sequence.