The fall armyworm (FAW) Spodoptera frugiperda (J.E. Smith) is a highly damaging invasive omnivorous pest that has developed varying degrees of resistance to commonly used insecticides. To investigate the molecular mechanisms of tolerance to tetraniliprole, spinetoram, and emamectin benzoate, the enzyme activity, synergistic effect, and RNA interference were implemented in S. frugiperda. The functions of cytochrome P450 monooxygenase (P450) in the tolerance to tetraniliprole, spinetoram, and emamectin benzoate in S. frugiperda was determined by analysing changes in detoxification metabolic enzyme activity and the effects of enzyme inhibitors on susceptibility to the three insecticides. 102 P450 genes were screened via transcriptome and genome, of which 67 P450 genes were differentially expressed in response to tetraniliprole, spinetoram, and emamectin benzoate and validated by quantitative real-time PCR. The expression patterns of CYP9A75, CYP340AA4, CYP340AX8v2, CYP340L16, CYP341B15v2, and CYP341B17v2 were analysed in different tissues and at different developmental stages in S. frugiperda. Silencing CYP340L16 significantly increased the susceptibility of S. frugiperda to tetraniliprole, spinetoram, and emamectin benzoate. Furthermore, knockdown of CYP340AX8v2, CYP9A75, and CYP341B17v2 significantly increased the sensitivity of S. frugiperda to tetraniliprole. Knockdown of CYP340AX8v2 and CYP340AA4 significantly increased mortality of S. frugiperda to spinetoram. Knockdown of CYP9A75 and CYP341B15v2 significantly increased the susceptibility of S. frugiperda to emamectin benzoate. These results may help to elucidate the mechanisms of tolerance to tetraniliprole, spinetoram and emamectin benzoate in S. frugiperda.

Panonychus citri is one of the most destructive pests in citrus orchards, exhibiting varying degrees of tolerance to numerous insecticides, such as spirodiclofen. To effectively manage pests, this study explores the response of P. citri to spirodiclofen stress from the perspectives of life history, enzymatic parameters, and reproduction. The effects of two concentrations (LC30 and LC50) of spirodiclofen on the biological parameters of P. citri were evaluated by the life table method. The results showed that the development duration, fecundity, oviposition days, and lifespan were shortened, though the pre-oviposition period of two treatments was prolonged in comparison with the control. A significant decrease was recorded in the net reproductive rate (R0) and the mean generation time (T) for the two treatments. Nevertheless, the intrinsic rate of increase (r) and the rate of increase (λ) were not significantly affected in the LC30 treatment, whereas they declined in the LC50 treatment. The enzyme activity assay resulted in higher activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and carboxylesterase (CarE), among the treatments than the control. In contrast, the treatments recorded lower cytochromeP450 (CYP450) and Glutathione S-transferase (GST) activities than the control. Furthermore, the study detected that relative mRNA expression of Vitellogenin (Vg) and Vitellogenin receptor (VgR) for two treatments were lower than the control. In summary, two concentrations of spirodiclofen inhibited progeny growth and fecundity of P. citri. Additionally, the results of this study may support further research on tolerance of P. citri in response to spirodiclofen stress.

Pentastiridius leporinus (Hemiptera: Cixiidae) is the main vector of an emerging and fast spreading sugar beet disease, the syndrome ‘basses richesses’ (SBR), in different European countries. The disease is caused by the γ-3-proteobacterium ‘Candidatus Arsenophonus phytopathogenicus’ and the phytoplasma ‘Candidatus Phytoplasma solani’ which are exclusively transmitted by planthoppers and can lead to a significant loss of sugar content and yield. Monitoring of this insect vector is important for disease management. However, the morphological identification is time consuming and challenging as two additional cixiid species Reptalus quinquecostatus and Hyalesthes obsoletus with a very close morphology have been reported in sugar beet fields. Further, identification of females and nymphs of P. leporinus at species level based on taxonomic key is not possible. In this study, an isothermal nucleic acid amplification based on recombinase polymerase amplification (RPA) was developed to specifically detect P. leporinus. In addition, real-time RPA was developed to detect both adults (male and female) and nymph stages using pure or crude nucleic acid extracts. The sensitivity of the real-time RPA for detection of P. leporinus was comparable to real-time PCR, but a shorter time (< 7 min) was required. This is a first report for real-time RPA application for P. leporinus detection using crude nucleic acid templates which can be applied for fast and specific detection of this vector in the field.

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.

Plant–soil interactions have bottom–up and top–down effects within a plant community. Heavy metal pollution can change plant–soil interactions, directly influence bottom–up effects and indirectly affect herbivores within the community. In turn, herbivores can affect plant–soil interactions through top–down effects. However, the combined effects of heavy metals and herbivores on soil enzymes, plants and herbivores have rarely been reported. Therefore, the effects of lead (Pb), Spodoptera litura and their combined effects on soil enzyme activities, pakchoi nutrition, defence compounds and S. litura fitness were examined here. Results showed that Pb, S. litura and their combined effects significantly affected soil enzymes, pakchoi and S. litura. Specifically, exposure to double stress (Pb and S. litura) decreased soil urease, phosphatase and sucrase activities compared with controls. Furthermore, the soluble protein and sugar contents of pakchoi decreased, and the trypsin inhibitor content and antioxidant enzyme activity increased. Finally, the S. litura development period was extended, and survival, emergence rates and body weight decreased after exposure to double stress. The combined stress of Pb and S. litura significantly decreased soil enzyme activities. Heavy metal accumulation in plants may create a superposition or synergistic effect with heavy metal-mediated plant chemical defence, further suppressing herbivore development. Pb, S. litura and their combined effects inhibited soil enzyme activities, improved pakchoi resistance and reduced S. litura development. The results reveal details of soil–plant–herbivore interactions and provide a reference for crop pest control management in the presence of heavy metal pollution.

Trilocha varians is one of the major pests of Ficus spp. Based on 19 bioclimatic variables provided by the Worldclim, our study analysed the suitable distribution areas of T. varians under current and future climate changes (SSP1-2.6, SSP2-4.5, SSP5-8.5) for two periods (the 2050s and 2090s) using the maximum entropy algorithm (MaxEnt) model. Key environmental variables affecting the geographic distribution of T. varians were also identified, and the changes in the area of suitable range under current and future climate changes were compared. The results showed that the key environmental variables affecting the distribution of T. varians were temperature and precipitation, comprising annual mean temperature (bio1), temperature seasonality (standard deviation × 100) (bio4), precipitation of driest month (bio14), and precipitation of driest quarter (bio17). Under the current climatic conditions, the suitable distribution area of T. varians is within the range of 92°13′E–122°08′E, 18°17′N–31°55′N. The current high, medium, and low suitable areas for T. varians predicted by the MaxEnt model are 14.00 × 104, 21.50 × 104, and 71.95 × 104 km2, of which the high suitable areas are mainly distributed in southern Guangdong, southwestern Guangxi, western Taiwan, Hong Kong, and Hainan. Under different future climatic conditions, some of the high, medium, and low suitability zones for T. varians increased and some decreased, but the mass centre did not migrate significantly. The Pearl River Basin is predicted to remain the main distribution area of T. varians.

Micromelalopha troglodyta (Graeser) is an important pest of poplar in China, and glutathione S-transferase (GST) is an important detoxifying enzyme in M. troglodyta. In this paper, three full-length GST genes from M. troglodyta were cloned and identified. These GST genes all belonged to the epsilon class (MtGSTe1, MtGSTe2, and MtGSTe3). Furthermore, the expression of these three MtGSTe genes in different tissues, including midguts and fat bodies, and the MtGSTe expression in association with different concentrations of tannic acid, including 0.001, 0.01, 0.1, 1, and 10 mg ml−1, were analysed in detail. The results showed that the expression levels of MtGSTe1, MtGSTe2, and MtGSTe3 were all the highest in the fourth instar larvae; the expression levels of MtGSTe1 and MtGSTe3 were the highest in fat bodies, while the expression level of MtGSTe2 was the highest in midguts. Furthermore, the expression of MtGSTe mRNA was induced by tannic acid in M. troglodyta. These studies were helpful to clarify the interaction between plant secondary substances and herbivorous insects at a deep level and provided a theoretical foundation for controlling M. troglodyta.

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.

Carboxylesterases (CarEs) is an important detoxification enzyme system in phase Ⅰ participating in insecticides resistance. In our previous study, SlCarE054, a CarEs gene from lepidoptera class, was screened out to be upregulated in a pyrethroids and organophosphates resistant population. Its overexpression was verified in two field-collected populations of Spodoptera litura (Lepidoptera: Noctuidae) resistant to pyrethroids and organophosphates by qRT-PCR. Spatiotemporal expression results showed that SlCarE054 was highly expressed in the pupae stage and the digestive tissue midgut. To further explore its role in pyrethroids and organophosphates resistance, its metabolism activity to insecticides was determined by UPLC. Its recombinant protein showed significant metabolism activity to cyhalothrin and fenvalerate, but not to phoxim or chlorpyrifos. The metabolic activity of SlCarE054 to β-cypermethrin showed stereoselectivity, with higher metabolic activity to θ-cypermethrin than the enantiomer α-cypermethrin. The metabolite of β-cypermethrin was identified as 3-phenoxybenzaldehyde. Further modelling and docking analysis indicated that β-cypermethrin, cyhalothrin and fenvalerate could bind with the catalytic triad of the 3D structure of SlCarE054. The interaction of β-cypermethrin with SlCarE054 also showed the lowest binding energy. Our work provides evidence that SlCarE054 play roles in β-cypermethrin resistance in S. litura.

Two-spotted spider mite, Tetranychus urticae Koch (Acari: Prostigmata), is one of the most economically important mite species, mainly controlled by chemical acaricides. Natural enemies have been assessed as reliable alternatives for management of this phytophagous mite. In the current project, demographic characteristics of Neoseiulus californicus McGregor (Acari: Phytoseiidae) to sublethal concentrations (LC10 = 6.76 × 102, LC20 = 8.74 × 103 and LC30 = 55.38 × 103 conidia ml−1) of entomopathogenic fungus, Beauveria bassiana (Bals.) Vuill. TV strain were investigated under laboratory conditions at 25 ± 2°C, 70 ± 5% RH and a photoperiod of 16:8 (L:D) h. Our results indicated that when adult predators were exposed to LC20 and LC30 of B. bassiana, the oviposition period was significantly reduced compared with other treatments. Neoseiulus californicus fecundity was significantly greater in the control (37 eggs) than in LC30 (24 eggs). Life table analysis revealed that the net reproductive rate (R0) declined as the sublethal concentrations of B. bassiana increased. The most striking result to emerge from the data is that not only intrinsic (r); but also, finite rate of increase (λ) was not significantly affected by different concentrations of B. bassiana. Our findings revealed some potential interactions of B. bassiana and N. californicus during their combinations for managing T. urticae that may be helpful for optimising control of this important pest.

The impact of invasive species on biodiversity, food security and economy is increasingly noticeable in various regions of the globe as a consequence of climate change. Yet, there is limited research on how climate change affects the distribution of the invasive Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera:Liviidae) in Ghana. Using maxnet package to fit the Maxent model in R software, we answered the following questions; (i) what are the main drivers for D. citri distribution, (ii) what are the D. citri-specific habitat requirements and (iii) how well do the risk maps fit with what we know to be correctly based on the available evidence?. We found that temperature seasonality (Bio04), mean temperature of warmest quarter (Bio10), precipitation of driest quarter (Bio17), moderate resolution imaging spectroradiometer land cover and precipitation seasonality (Bio15), were the most important drivers of D. citri distribution. The results follow the known distribution records of the pest with potential expansion of habitat suitability in the future. Because many invasive species, including D. citri, can adapt to the changing climates, our findings can serve as a guide for surveillance, tracking and prevention of D. citri spread in Ghana.

Drosophila suzukii (Matsumura) is an exotic pest of economic importance that affects several soft-skinned fruits in Mexico. Previously, we found that yellow or yellow-green rectangular cards inside a transparent trap baited with attractants improved D. suzukii capture. In this study, we evaluated the influence of rectangular cards with different yellow shades inside a transparent multi-hole trap baited with apple cider vinegar (ACV) on D. suzukii capture in the field. Second, we tested whether ACV-baited traps with cards of other geometric shapes affected D. suzukii catches compared to traps with rectangular cards. Third, we evaluated the effects of commercial lures combined with a more efficient visual stimulus from previous experiments on trapping D. suzukii flies. We found that ACV-baited traps plus a yellow-shaded rectangle card with 67% reflectance at a 549.74 nm dominant wavelength captured more flies than ACV-baited traps with yellow rectangle cards with a higher reflectance. Overall, ACV-baited traps with rectangles and squares caught more flies than did ACV-baited traps without visual stimuli. The traps baited with SuzukiiLURE-Max, ACV and Z-Kinol plus yellow rectangles caught 57, 70 and 101% more flies, respectively, than the traps baited with the lure but without a visual stimulus.

Anthropogenic-mediated climate change is expected to negatively affect pest management in agriculture. Hence, we investigated the oviposition, immature mortality, and developmental processes of Spodoptera species (Spodoptera exigua (Hübner) and Spodoptera litura (Fabricius)) under different temperatures (20, 25, and 30°C) and relative humidity (RH) (30–35, 50–55, 70–75, and 90–95%) conditions. For fecundity, mouths of each Spodoptera species were released into a rectangular box whose inner walls were covered with a sheet of white paper for each combination of temperature and RH. The mouths were kept inside the box to deposit eggs for 72 h. Temperature and RH significantly affected the fecundity, with the maximum number of eggs laid in 70–75% at 30°C. The highest egg and larval mortalities were recorded in 30–35 and 90–95% RH, respectively. Temperature and RH greatly affected the developmental period (egg–adult) and adult emergence rate. The rapid development was recorded in 70–75% RH at 30°C. Higher number of adults was found with an increase in temperature and RH. Adult longevity was significantly higher in 70–75% RH at 20°C. Based on the present study's findings, temperature and RH had an individual apparent effect on the developmental processes of Spodoptera species instead of an interactive effect. Therefore, there is need for an in-depth study of the influence of several climatic factors, including CO2, on the developmental modality and demographic changes of Spodoptera species to assess the impacts of climatic components and the sustainable development of management strategies.

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.

MicroRNAs (miRNAs) are endogenous, non-coding RNAs, which are functional in a variety of biological processes through post-transcriptional regulation of gene expression. However, the role of miRNAs in the interaction between Bacillus thuringiensis and insects remains unclear. In this study, small RNA libraries were constructed for B. thuringiensis-infected (Bt) and uninfected (CK) Spodoptera exigua larvae (treated with double-distilled water) using Illumina sequencing. Utilising the miRDeep2 and Randfold, a total of 233 known and 726 novel miRNAs were identified, among which 16 up-regulated and 34 down-regulated differentially expressed (DE) miRNAs were identified compared to the CK. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that potential target genes of DE miRNAs were associated with ABC transporters, fatty acid metabolism and MAPK signalling pathway which are related to the development, reproduction and immunity. Moreover, two miRNA core genes, SeDicer1 and SeAgo1 were identified. The phylogenetic tree showed that lepidopteran Dicer1 clustered into one branch, with SeDicer1 in the position closest to Spodoptera litura Dicer1. A similar phylogenetic relationship was observed in the Ago1 protein. Expression of SeDicer1 increased at 72 h post infection (hpi) with B. thuringiensis; however, expression of SeDicer1 and SeAgo1 decreased at 96 hpi. The RNAi results showed that the knockdown of SeDicer1 directly caused the down-regulation of miRNAs and promoted the mortality of S. exigua infected by B. thuringiensis GS57. In conclusion, our study is crucial to understand the relationship between miRNAs and various biological processes caused by B. thuringiensis infection, and develop an integrated pest management strategy for S. exigua via miRNAs.

Two fruit fly (Diptera: Tephritidae) species of economic importance: Ceratitis rosa Karsch and Ceratitis quilicii De Meyer, Mwatawala & Virgilio are present in South Africa. The two species were considered as one species prior to 2016, but were subsequently separated. In this study, the distribution and abundance of the two species were quantified in seven provinces in South Africa through trapping with Enriched Ginger Oil as an attractant. Trapping was conducted over three seasons across two years (2020 and 2021): late summer, autumn-winter, and spring-early summer. Host ranges of the two species were investigated by fruit sampling in and outside of trapping sites. Ceratitis quilicii was more widely distributed than C. rosa with the latter being recorded in only three north-eastern provinces. There were geographical limits for both species with no records of them in Northern Cape Province. Catches of C. quilicii were higher in summer with average temperatures varying from 15 to 27°C while for C. rosa, catches remained low and consistent between seasons. Ceratitis quilicii catches decreased at lower rates than those of C. rosa at temperatures below 15°C. The two species were reared from 13 plant species from nine families. Four of these hosts were infested by both C. quilicii and C. rosa in the same province where they occurred. Preferred hosts of the two species belonged to the Myrtaceae family. The characterisation of the distribution, abundance and host ranges of these pests will provide a baseline for pest status determination and implementation of management actions.

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.

Gene expression studies in organisms are often conducted using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and the accuracy of RT-qPCR results relies on the stability of reference genes. We examined ten candidate reference genes in Sclerodermus guani, a parasitoid wasp that is a natural enemy of long-horned beetle pests in forestry, including ACT, EF1α, Hsc70, Hsp70, SRSF7, α-tubulin, RPL7A, 18S, 28S, and SOD1, regarding variable biotic and abiotic factors such as body part, life stage, hormone, diet, and temperature. Data were analysed using four dedicated algorithms (ΔCt, BestKeeper, NormFinder, and geNorm) and one comparative tool (RefFinder). Our results showed that the most stable reference genes were RPL7A and EF1α regarding the body part, SRSF7 and Hsc70 regarding the diet, RPL7A and α-tubulin regarding the hormone, SRSF7 and RPL7A regarding the life stage, and SRSF7 and α-tubulin regarding temperature. To ascertain the applicability of specific reference genes, the expression level of the target gene (ACPase) was estimated regarding the body part using the most stable reference genes, RPL7A and EF1α, and the least stable one, SOD1. The highest expression level of ACPase was observed in the abdomen, and the validity of RPL7A and EF1α was confirmed. This study provides, for the first time, an extensive list of reliable reference genes for molecular biology studies in S. guani.

Reticulitermes chinensis Snyder is an important pest in forestry and construction and is widely distributed in China. We found that Serratia marcescens Bizio strain SM1 has insecticidal activity to R. chinensis, but the pathogenic mechanism of SM1 to R. chinensis is not clear. Therefore, full-length transcriptome sequencing was performed on R. chinensis infected with SM1 and the control group. A total of 230 differentially expressed genes were identified by comparing SM1 infection group and the control group, among which 103 were downregulated and 127 were upregulated. We found downregulated genes in nine metabolic pathway categories, among which carbohydrate metabolism had the most downregulated genes, followed by energy metabolism and amino acid metabolism. We also found that some downregulated genes were related to pattern recognition receptors, cellular immunity, and humoral immunity, indicating that R. chinensis immunity was negatively affected by SM1 infection. In addition, some genes in signal transduction and genetic information processing pathways were downregulated. In this study, high-throughput full-length transcriptome analysis was used to analyse the pathogenic mechanism of SM1 to R. chinensis. The results of this study provide useful information for exploring the relationship between SM1 and R. chinensis, and provide theoretical support for the future application of SM1 and the prevention and treatment of R. chinensis.