Spotted-wing drosophila, Drosophila suzukii (Matsumura), is an invasive vinegar fly that is a major threat to the small fruits industries globally. Insect capa genes encode multiple neuropeptides, including CAPA-periviscerokinin (CAPA-PVK) peptides, that are specifically known to cause diuresis or anti-diuresis in various organisms. Here we identified and characterized a corresponding G protein-coupled receptor (GPCR) of the D. suzukii CAPA-PVK peptides: CAPA receptor (CAPA-R). To better characterize the behavior of D. suzukii CAPA-R, we used insect cell-based functional expression assays to evaluate responses of CAPA-R against D. suzukii CAPA-PVKs, CAPA-PVKs from five species in Insecta, one species from Mollusca, modified CAPA-PVK peptides, and some PRXamide family peptides: pyrokinin (PK), diapause hormone (DH), and ecdysis-triggering hormone (ETH). Functional studies revealed that the D. suzukii CAPA-R is strongly activated by both of its own natural D. suzukii CAPA-PVKs, and interestingly, it was strongly activated by other CAPA-PVK peptides from Frankliniella occidentallis (Thysanoptera), Solenopsis invicta (Hymenoptera), Helicoverpa zea (Lepidoptera) and Plutella xylostella (Lepidoptera). However, D. suzukii CAPA-R was not activated by Mollusca CAPA-PVK or the other PRXamide peptides. Gene expression analyses showed that the CAPA-R was highly expressed in the Malpighian tubules and moderately in hindgut compared to other digestive organs or the rest of body, supporting diuretic/antidiuretic functionality. When compared across life stages of D. suzukii, expression of CAPA-R was approximately 1.5x greater in the third instar than the other stages and minimally detected in the eggs, 4-day old pupae and 3-day old adults. Our results functionally characterized the D. suzukii CAPA-R and a few short peptides were identified as potential biological targets to exploit the CAPA-R for D. suzukii management.

Megalurothrips usitatus Bagnall, an important pest of bean plants, is primarily managed with synthetic insecticides. M. usitatus has developed considerable resistance to various insecticides in multiple cowpea-growing areas in Hainan Province, China, posing challenges to its control in the field. Light control technology is a potentially effective physical control method for M. usitatus. The vision of thrips is highly sensitive to UV light, whereas other biological characteristics remain unknown. Therefore, this study evaluated the effects of ultraviolet light on the biological characteristics of M. usitatus. Results showed that the egg, larval, and pupal stages of M. usitatus were significantly shortened, and the emergence rate (79.59%) and adult survival rate (77.95%) were reduced under a devoid of UV light environment (UV-), compared with the full-spectrum light (control treatment group, CK) (p < 0.05). However, the single spawning quantity and total amount of spawning were significantly higher, and the sex ratio (57%) was the highest under UV- (p < 0.05). Single UV light (UV+) only affected the pupation rate. Also, the antioxidant enzymes, polyphenol oxidase, superoxide dismutase (SOD), and peroxidase activities were significantly and negatively correlated with the progression of generations under UV-, whereas catalase and SOD activities were significantly and positively correlated with the progression of generations under UV+. The UV- light conditions significantly interfered with the behavior selection of M. usitatus. The results of this study showed that the adaptability of M. usitatus populations would be greatly reduced in the absence of ultraviolet light, providing a theoretical basis for the control of M. usitatus populations.

Several species of the worldwide distributed genus Calliphora Robineau-Desvoidy (Insecta, Diptera, Calliphoridae) are medically important vectors and agents of myiasis. Furthermore, these flies are relevant in forensics because they are found in corpses. Information regarding the taxonomy, bionomics and distribution of Calliphora species endemic to South America, including Calliphora lopesi Mello, is scarce. To expand knowledge on C. lopesi, this study presents descriptions of eggs, larvae, puparia and developmental data at 14, 17, 20, 23 and 26 ± 1°C for the first time. Adult flies were collected from the field and kept in the laboratory to obtain samples for morphological and biological studies. Immatures were examined using light and scanning electron microscopy. To assess the growth rate, 10 specimens from each temperature group were randomly removed from the diet and weighed every 24 h from larval hatching until pupation. The minimum developmental threshold, thermal constant and linear development-rate equations were calculated for each stage. Considering weight gain records and survival rates, the optimum temperature for the development of C. lopesi ranges from 23 to 26°C. A key to third-instar larvae of known Neotropical species of Calliphora was also provided to assist in identification. The information provided in this study should be useful in expanding knowledge about Neotropical Calliphoridae species of forensic importance.
Várias espécies do gênero Calliphora Robineau‐Desvoidy (Insecta, Diptera, Calliphoridae), distribuídas mundialmente, são vetores e agentes causadoras de miíases clinicamente importantes. Além disso, tais moscas são relevantes no âmbito forense porque são encontradas em cadáveres. Informações sobre a taxonomia, bionomia e distribuição de espécies de Calliphora endêmicas da América do Sul, incluindo Calliphora lopesi Mello, ainda são escassas. Para ampliar o conhecimento sobre a espécie C. lopesi, este estudo apresenta pela primeira vez as descrições de ovos, larvas e pupários, assim como dados sobre o desenvolvimento de imaturos a 14, 17, 20, 23 e 26 ± 1°C. Moscas adultas foram coletadas no campo e mantidas em laboratório para obtenção de amostras para estudos morfológicos e biológicos. Os imaturos foram examinados utilizando microscopia óptica e eletrônica de varredura. Para avaliar a taxa de crescimento, 10 espécimes de cada grupo de temperatura foram retirados aleatoriamente do substrato em que se alimentavam e pesados a cada 24 horas, desde a eclosão das larvas até a pupariação. O limiar mínimo de desenvolvimento, a constante térmica e as equações lineares da taxa de desenvolvimento foram calculados para cada estágio. Levando em consideração os registros de ganho de peso e taxas de sobrevivência, a temperatura ótima para o desenvolvimento de C. lopesi varia de 23 a 26°C. Uma chave para larvas de terceiro estádio de espécies neotropicais conhecidas de Calliphora também está sendo disponibilizada para auxiliar na identificação. Espera‐se que as informações fornecidas neste estudo possam ser úteis para ampliar o conhecimento sobre espécies Neotropicais de Calliphoridae de importância forense.

BACKGROUND: To assess whether the \'economic boom\' in the tropical seaport city of Barranquilla improved tapped water supplies to socio-economically poor neighbourhoods resulting in: (1) their reduced use for domestic water-storage in large (> 1,000-litre) custom-made cement tanks which are their principal Aedes aegypti breeding sites and (2) their pupae/person index (PPI) values to below their established 0.5-1.5 PPI arbovirus transmission-threshold value, compared to matched neighbourhoods in the: (a) pre-economic boom (2004) period in Barranquilla and (b) economically-neglected seaport city of Buenaventura.
METHODS: The simple, accurate and robust water surface sweep-net/calibration factor or total count methods were used to determine the total Ae. aegypti pupae numbers in greater or less than 20-litre water-holding container types located \'inside\' or \'outside\' these neighbourhood premises. The women residents also participated in questionnaire-based responses about their domestic water supplies, water-storage and maintenance and mosquito life stages and disease transmission knowledge, to subsequently plan appropriate resident education programmes. Microsoft Excel 8.0 with OpenEpi was used to determine the samples sizes and the statistical values.
RESULTS: Tapped water supplies to the three poor Barranquilla neighbourhoods were dramatically increased from 2004 to 2023 resulting in their residents significantly reducing their: (a) large cement water-storage tanks from 1 per 6.9 (2004) to 1 per 31.2 (2020) premises (z = 10.5: p = 0) and (b) PPI values to 0.16, 0.19 and 0.53 (mean: 0.29: 95% CI ± 0.4) in each study neighbourhood. In contrast, tapped water supplies remained inadequate in the Buenaventura neighborhoods, thereby resulting in their continued use of many large (> 1,000-litre) water-storage containers (Barranquilla: 1 per 31.2 and Buenaventura: 1 per 1.5 premises: z = - 9.26: p = 0), with unacceptably high 0.81, 0.88 and 0.99 PPI values in each study neighbourhood (mean 0.89: 95% CI ± 0.12).
CONCLUSIONS: Improved tapped water supplies resulted in reduced numbers of large custom-made stoneware water-containers, as are employed by poor residents throughout the world, as well as their Ae. aegypti PPI transmission threshold values which, together with appropriate residents\' education programmes, are also urgently to reduce to prevent/reduce Ae. aegypti transmitted human diseases globally.

In holometabolous insects, the larval body is almost completely decomposed and reconstructed into the adult body during the pupal-pharate adult stages. Therefore, the total energetic cost of this process is a key thermodynamic quantity necessary for evaluating the benefit of their life history. Here, we measured whole-body thermal dissipation of single pupae of the fruit fly, Drosophila melanogaster, during the period from puparium formation to adult eclosion as a function of age, using a high-precision isothermal calorimeter at T = 298 K. The mass-specific energy consumption during the period from the onset of larval-pupal apolysis to adult eclosion was determined to be 2.3 kJ/g for an individual of mass (adult) = 1.0 mg, while it was observed to follow Kleiber\'s law for individuals smaller than mass (adult) = 1.0 mg. During the pupal-pharate adult period, in addition to the U-shaped variation, several characteristic thermal dissipations related to various events, including somatic muscle contractions, ecdyses, pulsatile hormone secretion in a pharate adult, and vaporization of the exuvial fluid, were observed. The periodic bursts in the pharate adult stage grew exponentially, suggesting that the positive feedback in the metabolic system synchronized with the progression of development, making the energy consumption in this stage more efficient. The present study showed that high-precision calorimetry is a powerful and credible method for measuring not only the total energy spent during development but also the energy spent during every specific developmental event in an organism.

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.

Effective mosquito population suppression has been repeatedly demonstrated in field trials through the release of male mosquitoes to induce sterile mating with wild females using the incompatible insect technique (IIT), the sterile insect technique (SIT), or their combination. However, upscaling these techniques requires a highly efficient and scalable approach for the sex separation of mass-reared mosquitoes to minimize the unintentional release of females, which can lead to either population replacement or biting nuisance, a major bottleneck up to now. Here, we report the successful development of an automated mosquito pupa sex sorter that can effectively separate large numbers of males from females for population suppression of Aedes aegypti, A. albopictus, and Culex quinquefasciatus. The male production capacity of the automated sex sorter was increased by ~17-fold compared with manual sex separation with the Fay-Morlan sorter and enabled one person to separate 16 million males per week. With ~0.5% female contamination, the produced males exhibited high flight ability and mating performance. The field trial demonstrates that the quality of A. albopictus males produced using the automated sex sorter is suitable for inducing population suppression. These results indicate that the automated sex sorter offers the potential to upscale IIT and SIT against mosquito vectors for disease control.

Butterflies constitute approximately 10% of lepidopteran insects, and along with silkworms, they can produce silk; however, this feature is often ignored. In the present study, we observed two primary methods used by butterflies to hang pupae on trees using silk: pupa adheraena (Danaus chrysippus) and pupa contigua (Papilio polytes). Anchoring the abdominal ends of pupae with a silk pad was the most common method used in both cases, whereas wrapping silk around the body using a silk girdle was a method unique to pupa contigua. The connection between the cremaster and silk pad was observed to be similar to that between the hook and loop of a Velcro fastener, except that the cremaster hook is anchor-shaped rather than being a single hook. Such a connection will remain secure, ensuring the safety of the pupae during exposure to wind and rain. Through determining the mechanical properties of silk, the performance of butterfly silk was found to be weaker than that of silkworm silk. Therefore, the P. polytes silk girdle adopts the strategy of merging a dozen silk threads to improve its strength and toughness, thereby making it difficult to break. In addition, we explained how the protein sequence and structure of butterfly silk impact its performance. In conclusion, we discovered that butterfly pupae develop unique body features to establish secure bonds with silk. This enables them to effectively undergo metamorphosis and endure harsh weather conditions and surroundings.

The weathering time of empty puparia could be important in predicting the minimum postmortem interval (PMImin). As corpse decomposition progresses to the skeletal stage, empty puparia often remain the sole evidence of fly activity at the scene. In this study, we used empty puparia of Sarcophaga peregrina (Diptera: Sarcophagidae) collected at ten different time points between January 2019 and February 2023 as our samples. Initially, we used the scanning electron microscope (SEM) to observe the surface of the empty puparia, but it was challenging to identify significant markers to estimate weathering time. We then utilized attenuated total internal reflectance Fourier transform infrared spectroscopy (ATR-FTIR) to detect the puparia spectrogram. Absorption peaks were observed at 1064 cm-1, 1236 cm-1, 1381 cm-1, 1538 cm-1, 1636 cm-1, 2852 cm-1, 2920 cm-1. Three machine learning models were used to regress the spectral data after dimensionality reduction using principal component analysis (PCA). Among them, eXtreme Gradient Boosting regression (XGBR) showed the best performance in the wavenumber range of 1800-600 cm-1, with a mean absolute error (MAE) of 1.20. This study highlights the value of refining these techniques for forensic applications involving entomological specimens and underscores the considerable potential of combining FTIR and machine learning in forensic practice.

Silkworms have been farmed for their silk since ancient times. After silk reeling, their chrysalides are consumed as food in several Asian countries. Despite the long rearing tradition of this insect, few studies have investigated the silkworm\'s microbiological safety all along the life cycle, focusing on detecting silkworm pathogens or on the safety of the dried chrysalis for food consumption. However, the in-farm rearing process, which takes around forty days, may affect the microbial load of the silkworm and of the rearing environment, as well as the quality of fresh cocoon and other performance parameters. No data is available on how microbial contamination changes during the rearing period and between different farmers. Furthermore, in light of the possible use of the chrysalis as food, it is crucial to understand how its microbial load varies according to the water content. To address these specific questions, we conducted an investigation involving the analysis of specific microbial indicators commonly used in the food chain. We collected environmental and silkworm samples from several farms. The examination covered the entire life cycle of silkworms, beginning with the first instar larvae and concluding with the scrutiny of both freshly harvested and dried pupae. Silkworm farms in Northeast Italy proved to be an appropriate model system for carrying out the experimentation. Additionally, an evaluation of rearing performance was conducted, with a focus on the quality of fresh cocoons and the survival rate of the insects.