Temperature is among the key factors impacting the establishment and spread of invasive pests. The tomato leafminer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is one of the major pests attacking Solanaceae plants and is known to possess overwintering capacities. However, the cold hardiness of T. absoluta pupae is poorly documented. In this study, we investigated the effect of constant temperature and stepwise cooling on T. absoluta pupae under laboratory conditions. For this purpose, bioassays on pupal development under constant temperature (5°C) for 30, 60 and 90 days, and stepwise changes in temperature (11, 10 and 8°C; in this order every 30 days), were assessed. We found that exposure to 5°C for 30 and 60 days did not affect the post-cooling emergence time of adults compared to the control. Pupae completed their development after 60 days of cold exposure at 5°C, but more adults emerged after 30 than 60 days. Even though alive pupae were observed after 90 days of cold exposure at 5°C, no adults emerged. External colours of pupae depended on the duration of cold periods, and green pupae obtained after 30 and 60 days were found to be positively correlated with the emergence of adults. When pupae were kept at 11°C for 30 days, 47% emerged, and when the temperature was changed to 10, only 12% of pupae emerged for the period 31–60 days. However, the decrease of the temperature to 8°C yielded no emergence for the period 61–90 days. Our study provides useful information to better understand the population dynamics of overwintering T. absoluta, and to underpin the development of monitoring and control strategies for the pest.

Mosquito-borne disease is a significant public health issue and within Australia Ross River virus (RRV) is the most reported. This study combines a mechanistic model of mosquito development for two mosquito vectors; Aedes vigilax and Aedes camptorhynchus, with climate projections from three climate models for two Representative Concentration Pathways (RCPs), to examine the possible effects of climate change and sea-level rise on a temperate tidal saltmarsh habitat in Perth, Western Australia. The projections were run under no accretion and accretion scenarios using a known mosquito habitat as a case study. This improves our understanding of the possible implications of sea-level rise, accretion and climate change for mosquito control programmes for similar habitats across temperate tidal areas found in Southwest Western Australia. The output of the model indicate that the proportion of the year mosquitoes are active increases. Population abundances of the two Aedes species increase markedly. The main drivers of changes in mosquito population abundances are increases in the frequency of inundation of the tidal wetland and size of the area inundated, increased minimum water temperature, and decreased daily temperature fluctuations as water depth increases due to sea level changes, particularly under the model with no accretion. The effects on mosquito populations are more marked for RCP 8.5 when compared to RCP 4.5 but were consistent among the three climate change models. The results indicate that Ae. vigilax is likely to be the most abundant species in 2030 and 2050, but that by 2070 Aedes camptorhynchus may become the more abundant species. This increase would put considerable pressure on existing mosquito control programmes and increase the risk of mosquito-borne disease and nuisance biting to the local community, and planning to mitigate these potential impacts should commence now.

Functional response and mutual interference are important attributes of natural enemies that should be analysed in species with the potential to be used as biological control agents in order to increase the predictive power of the possible benefits and/or consequences of their release in the field. Our main objective was to determine the functional response and mutual interference of Coptera haywardi (Oglobin), a pupal parasitoid of economically important fruit flies (Diptera: Tephritidae). The functional response of C. haywardi on A. ludens pupae corresponded to a type II model, with an attack rate of 0.0134 host pupa/h and a handling time of 1.843 h, which reveals a meticulous selection process of pupal hosts. The effect of mutual interference among foraging females was negatively correlated with increased parasitoid density in the experimental arena, showing a gradual decline in attack rate per individual female. The increase in the number of foraging females also had an impact on the number of oviposition scars per pupa and the number of immature parasitoids per dissected pupa, but not on the percentage of adult emergence or the sex ratio. Our results suggest that C. haywardi could act as a complementary parasitoid in the control of fruit fly pupae, since the random distribution of these pupae in the soil would decrease the possibility of aggregation and mutual interference between foraging females.

Understanding the blood-feeding patterns of mosquitoes is essential for evaluating their potential as disease vectors, especially in urban areas where mosquitoes coexist with humans, domestic animals and wildlife. This study aimed to bridge a substantial gap in regional knowledge by identifying the blood meal sources of field-collected mosquitoes in domestic and open green environments from two urbanisations of temperate Argentina, the Área Metropolitana de Buenos Aires (AMBA) and Tandil, using molecular techniques. Female mosquitoes were collected from November 2019 to March 2020 and April–May 2021. A bipartite network analysis was performed for each environment and urbanisation. A total of 103 blood meals from Aedes (2 species) and Culex (7 species) were identified. Among these, five mammal and 18 bird species were recognised as hosts. Aedes mosquitoes exclusively fed on mammals, while Culex mosquitoes exhibited a broader host range including both birds and mammals. In AMBA, the open green environments were composed by more mosquito species than the domestic environments, while both presented similar numbers of vertebrate species. In contrast, in open green environments from Tandil only blood-fed Aedes albifasciatus were collected. For open green environments of AMBA and domestic environments of Tandil, results suggested some degree of host selection. For the three main vectors of diseases in the region, Aedes aegypti, Ae. albifasciatus and Culex pipiens molestus, we present the first molecular evidence of human blood meals in South America. Epidemiological significance of the present findings is discussed.

Hemiptera insects exhibit a close relationship to plants and demonstrate a diverse range of dietary preferences, encompassing phytophagy as the predominant feeding habit while a minority engages in carnivorous or haematophagous behaviour. To counteract the challenges posed by phytophagous insects, plants have developed an array of toxic compounds, causing significant evolutionary selection pressure on these insects. In this study, we employed a comparative genomics approach to analyse the expansion and contraction of gene families specific to phytophagous insect lineages, along with their adaptive evolutionary traits, utilising representative species from the Hemiptera order. Our investigation revealed substantial expansions of gene families within the phytophagous lineages, especially in the Pentatomomorpha branch represented by Oncopeltus fasciatus and Riptortus pedestris. Notably, these expansions of gene families encoding enzymes are potentially involved in hemipteran-plant interactions. Moreover, the adaptive evolutionary analysis of these lineages revealed a higher prevalence of adaptively evolved genes in the Pentatomomorpha branch. The observed branch-specific gene expansions and adaptive evolution likely contribute significantly to the diversification of species within Hemiptera. These results help enhance our understanding of the genomic characteristics of the evolution of different feeding habits in hemipteran insects.

Aphis spiraecola Patch is one of the most economically important tree fruit pests worldwide. The pyrethroid insecticide lambda-cyhalothrin is commonly used to control A. spiraecola. In this 2-year study, we quantified the resistance level of A. spiraecola to lambda-cyhalothrin in different regions of the Shaanxi province, China. The results showed that A. spiraecola had reached extremely high resistance levels with a 174-fold resistance ratio (RR) found in the Xunyi region. In addition, we compared the enzymatic activity and expression level of P450 genes among eight A. spiraecola populations. The P450 activity of A. spiraecola was significantly increased in five regions (Xunyi, Liquan, Fengxiang, Luochuan, and Xinping) compared to susceptible strain (SS). The expression levels of CYP6CY7, CYP6CY14, CYP6CY22, P4504C1-like, P4506a13, CYP4CZ1, CYP380C47, and CYP4CJ2 genes were significantly increased under lambda-cyhalothrin treatment and in the resistant field populations. A L1014F mutation in the sodium channel gene was found and the mutation rate was positively correlated with the LC50 of lambda-cyhalothrin. In conclusion, the levels of lambda-cyhalothrin resistance of A. spiraecola field populations were associated with P450s and L1014F mutations. Our combined findings provide evidence on the resistance mechanism of A. spiraecola to lambda-cyhalothrin and give a theoretical basis for rational and effective control of this pest species.

The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is a major destructive insect pest of coffee, which impacts the coffee crops negatively. As a draft genome has been completed for this insect, most molecular studies on gene transcriptional levels under different experimental conditions will be conducted using real-time reverse-transcription quantitative polymerase chain reactions (RT-qPCR). However, the lack of suitable internal reference genes will affect the accuracy of RT-qPCR results. In this study, the expression stability of nine candidate reference genes was evaluated under different developmental stages, temperature stress, and Beauveria bassiana infection. Data analyses were completed by four commonly used programs, BestKeeper, NormFinder, geNorm, and RefFinder. The result showed that RPL3 and EF1α combination were recommended as the most stable reference genes for developmental stages. EF1α and RPS3a combination were the top two stable reference genes for B. bassiana infection. RPS3a and RPL3 combination performed as the optimal reference genes both in temperature stress and all samples. Our results should provide a good foundation for the expression profile analyses of target genes in the future, especially for molecular studies on insect genetic development, temperature adaptability, and immune mechanism to entomogenous fungi in H. hampei.

From the 1990s, the Southeast Asia native giant resin bee Megachile sculpturalis (Smith, 1853) was introduced first to North America, and then to many countries in Europe. Despite increasing studies on its invasive potential and geographical expansion, information on nesting behaviour of this species is still extremely scarce. To increase knowledge on the nesting biology of M. sculpturalis, we studied multiple aspects of nesting and pollen provisioning in three consecutive years in artificial nests in Bologna, Italy. We observed 166 bees visiting nests, and followed individual nesting behaviour and success of 41 adult females. We measured cavity diameter in 552 nests and characterised the structure in 100 of them. More than 95% of nest diameters ranged between 0.6 and 1.2 cm, overlapping with several sympatric species of cavity-nesting hymenopterans in the study area. Most nests had a first chamber from the entrance of variable length without brood, followed by an average of about two brood cells with a mean length of 2.85 ± 0.13 cm each. The pollen stored in brood cells was almost monofloral, belonging to the ornamental plant Styphnolobium japonicum (L.) Schott. We estimated that a single female should visit ≈180 flowers to collect enough pollen for a single brood cell. These results fill knowledge gaps on the nesting biology and nest structure of the exotic M. sculpturalis, and they are discussed in relation to possible competition with native bees for nesting sites and foraging resources.

Bees play a significant role in the health of terrestrial ecosystems. The decline of bee populations due to colony collapse disorder around the world constitutes a severe ecological danger. Maintaining high yield of honey and understanding of bee behaviour necessitate constant attention to the hives. Research initiatives have been taken to establish monitoring programs to study the behaviour of bees in accessing their habitat. Monitoring the sanitation and development of bee brood allows for preventative measures to be taken against mite infections and an overall improvement in the brood's health. This study proposed a precision beekeeping method that aims to reduce bee colony mortality and improve conventional apiculture through the use of technological tools to gather, analyse, and understand bee colony characteristics. This research presents the application of advanced digital image processing with computer vision techniques for the visual identification and analysis of bee brood at various developing stages. The beehive images are first preprocessed to enhance the important features of object. Further, object is segmented and classified using computer vision techniques. The research is carried out with the images containing variety of immature brood stages. The suggested method and existing methods are tested and compared to evaluate efficiency of proposed methodology.

The European grapevine moth Lobesia botrana (Denis & Shiffermüller 1776) is an economically important pest of the vine-growing areas worldwide. Chemical insecticides have been used for its control; however, its resistance status is largely unknown in many regions. We monitored the susceptibility of several L. botrana populations from Greece and Turkey. In addition, based on RNAseq transcriptome analysis, we identified and phylogenetically classify the cytochrome P450 genes of L. botrana, as well as analysed target site sequences and looked for the presence of known resistance mutations. Resistance against chlorantraniliprole, alpha-cypermethrin, spinetoram, etofenprox, and acetamiprid was very low (below 2.5-fold in all cases, compared to a reference strain from Greece) in all populations from Greece that were included in the study. However, resistance against indoxacarb (4–30-fold), spinosad (5–59-fold), and deltamethrin (18–30 fold) was detected in the L. botrana populations from Turkey, compared to a reference population from Turkey. De novo transcriptome assembly and manual annotation, and subsequent PCR-based analysis of insecticide target sequences (i.e. voltage-gated sodium channel – VGSC: target of pyrethroids and oxadiazines; nicotinic acetylcholine receptor subunit a6 – nAChR_α6: target of spinosad; ryanodine receptor – RyR: target of diamides; glutamate-gated chloride channel – GluCl: target of avermectins and; acetylcholinesterase – AChE: target of organophosphates) showed the absence of known resistance mutations in all specimens from both countries. Finally, the L. botrana CYPome (116 genes) was manually analysed and phylogenetically characterised, to provide resources for future studies that will aim the analysis of metabolic resistance.

The gene encoding juvenile hormone response (Krüppel homolog1, Kr-hl) in Coccinella septempunctata was investigated by cloning and analysing expression profiles in different developmental stages and tissues by quantitative real-time polymerase chain reaction (PCR). C. septempunctata Kr-hl (CsKr-hl) encoded a 1338 bp open reading frame (ORF) with a predicted protein product of 445 amino acids; the latter showed high similarity to orthologs in other species and contained eight highly-conserved Zn-finger motifs for DNA-binding. CsKr-hl was expressed in different developmental stages of C. septempunctata. The expression levels of CsKr-hl in eggs, 2nd, 3rd, 4th instar larvae, and pupa were 3.31, 2.30, 7.09, 0.58, and 7.48 times the number of 1st instar larvae, respectively. CsKr-hl expression levels in female adults gradually increased at 25–30 days and were significantly higher than expression at 1–20 days. CsKr-hl expression in 20–30 days-old male adults was significantly higher than males aged 1–15 days. CsKr-hl expression levels in heads of male and female adults were significantly higher than expression levels in the thorax, adipose, and reproductive system. Interestingly, CsKr-hl expression levels in the adipose and reproductive system of female adults were significantly higher than in adult male corresponding organs, which suggest that CsKr-hl plays an important role in regulating reproductive development in C. septempunctata.

The subfamilies Salassinae and Agliinae are two monogeneric groups of the family Saturniidae. They were regarded as the non-cocooning saturniids in Asia. Since very little information on their life history and mitogenome has been reported, their origin and evolution are still poorly understood. In this study, nature-imitated rearing is used to record the life history of two Aglia and five Salassa species. In addition, four complete mitogenomes are presented, which are the first ones of these two subfamilies. The results show that both Salassinae and Agliinae have lost their cocooning. Moreover, the phylogenetic analysis demonstrates that the subfamily Saturniinae is not monophyletic due to the inclusion of Agliinae and Salassinae.

Neotropical cyclocephaline beetles, a diverse group of flower-loving insects, significantly impact natural and agricultural ecosystems. In particular, the genus Cyclocephala, with over 350 species, displays polymorphism and cryptic complexes. Lacking a comprehensive DNA barcoding framework, accessible tools for species differentiation are needed for research in taxonomy, ecology, and crop management. Moreover, cuticular hydrocarbons are believed to be involved in sexual recognition mechanisms in these beetles. In the present study we examined the cuticular chemical profiles of six species from the genus Cyclocephala and two populations of Erioscelis emarginata and assessed their efficiency in population, species, and sex differentiation. Overall we identified 74 compounds in cuticular extracts of the selected taxa. Linear alkanes and unsaturated hydrocarbons were prominent, with ten compounds between them explaining 85.6% of species dissimilarity. Although the cuticular chemical profiles efficiently differentiated all investigated taxa, only C. ohausiana showed significant cuticular profile differences between sexes. Our analysis also revealed two E. emarginata clades within a larger group of ‘Cyclocephala’ species, but they were not aligned with the two studied populations. Our research underscores the significance of cuticular lipid profiles in distinguishing selected cyclocephaline beetle species and contemplates their potential impact as contact pheromones on sexual segregation and speciation.

Aulacophora lewisii Baly (Coleoptera: Chrysomelidae) is an important pest of Luffa acutangula (L.) Roxb. (Cucurbitaceae) in India. Larvae of A. lewisii feed on the roots, while adults consume leaves of L. acutangula. In the current study, effects of three L. acutangula cultivars (Abhiskar, Debsundari, and Jaipur Long) on the life table parameters by age-stage, two-sex approach, and key digestive enzymatic activities (amylolytic, proteolytic, and lipolytic) of the larvae and adults of A. lewisii were determined. Further, nutrients (total carbohydrates, proteins, lipids, amino acids, and nitrogen content) and antinutrients (total phenols, flavonols, and tannins) present in the roots and leaves of three cultivars were estimated. The development time (egg to adult emergence) was fastest and slowest on Jaipur Long (31.80 days) and Abhiskar (40.91 days), respectively. Fecundity was highest and lowest on Jaipur Long (279.91 eggs) and Abhiskar (137.18 eggs), respectively. The intrinsic rate of increase (r) was lowest on Abhiskar (0.0511 day−1) and highest on Jaipur Long (0.0872 day−1). The net reproductive rate (R0) was lowest on Abhiskar (23.32 offspring female−1). The mean generation time (T) was shortest on Jaipur Long (52.59 days) and longest on Abhiskar (61.58 days). The amylolytic, proteolytic, and lipolytic activities of larvae and adults of A. lewisii were highest and lowest on Jaipur Long and Abhiskar, respectively. The lower level of nutrients and higher level of antinutrients influenced higher larval development time and lower fecundity of A. lewisii on Abhiskar than other cultivars. Our results suggest that Abhiskar cultivar could be promoted for cultivation.

Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), the cactus moth, is native to South America with a widespread distribution in Argentina. The larvae consume the interior of Opuntia spp. (Cactaceae) plants. The moth was used as a biocontrol agent against invasive non-native Opuntia spp. in many countries around the world. The cactus moth arrived unintentionally in Florida, USA, expanded its range and threatened Opuntia-based agriculture and natural ecosystems in southern North America. The insect is also a pest of cultivated O. ficus-indica L. in Argentina. An endemic South American parasitoid, Goniozus legneri Gordth (Hymenoptera: Bethylidae), is used in inundative biological control programmes against lepidopteran pests. The goal of this work was to evaluate G. legneri as a biocontrol agent to be used in inundative releases against C. cactorum. Mortality of C. cactorum by G. legneri was assessed at different spatial scales, as well as the interactions with Apanteles opuntiarum Martínez & Berta (Hymenoptera: Braconidae), a common Argentine natural enemy of C. cactorum. The ability of G. legneri to paralyse, parasitise and kill C. cactorum was confirmed. The paralysis inflicted on C. cactorum larvae reduced larval damage to the plants by 85%. Using two parasitoid species increased the mortality of C. cactorum larvae, but it was highly dependent on the order of their arrival. The combined mortality caused by both parasitoids was higher than a single one, in particular when G. legneri arrived first (56 ± 1%), suggesting asymmetric competition due to the preference of G. legneri attacking previously parasitised larvae. Goniozus legneri has potential as an inundative biocontrol agent of C. cactorum, but its interaction with the classical biocontrol agent A. opuntiarum needs to be considered.