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.

Many parasitoids alter their reproductive behaviour in response to the quality of encountered hosts. They make adaptive decisions concerning whether to parasitise a potential host, the number of eggs laid on an accepted host, and the allocation of sex to their offspring. Here we present evidence that Goniozus jacintae Farrugia (Hymenoptera: Bethylidae), a gregarious ectoparasitoid of larval tortricids, adjusts its reproductive response to the size and developmental stage of larvae of the light brown apple moth (LBAM), Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae). Goniozus jacintae parasitises instars 3–6 of LBAM, but most readily parasitises the later, larger, instars. Brood sizes were bigger on larger hosts and brood sex ratios were female biased (proportion of males = 0.23) with extremely low variance (never >1 male in a brood at emergence), perhaps the most precise of all studied bethylids. Host size did not influence brood development time, which averaged 19.64 days, or the body size of male offspring. However, the size of females was positively correlated with host size and negatively correlated with brood size. The sizes of individual males and females were positively related to the average amount of host resource available to individuals within each brood, suggesting that adult body size is affected by scramble competition among feeding larvae. Average brood sizes were: 3rd instar host, 1.3 (SE ± 0.075); 4th instar, 2.8 (SE ± 0.18); 5th instar, 4.7 (SE ± 0.23); 6th instar, 5.4 (SE ± 0.28). The largest brood size observed was 8 individuals (7 females, 1 male) on the 6th instar of LBAM. These results suggest that later instars would give the highest yield to optimise mass-rearing of G. jacintae if used for augmentative biological pest control.

The symbiosis between microorganisms and host arthropods can cause biological, physiological, and reproductive changes in the host population. The present study aimed to survey facultative symbionts of the genera Wolbachia, Arsenophonus, Cardinium, Rickettsia, and Nosema in Cotesia flavipes (Cameron) (Hymenoptera: Braconidae) and Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae) in the laboratory and evaluate the influence of infection on the fitness of these hosts. For this purpose, 16S rDNA primers were used to detect these facultative symbionts in the host species, and the hosts' biological and morphological features were evaluated for changes resulting from the infection caused by these microorganisms. The bacterial symbionts studied herein were not detected in the D. saccharalis samples analysed, but the endosymbiont Wolbachia was detected in C. flavipes and altered the biological and morphological aspects of this parasitoid insect. The results of this study may help to elucidate the role of Wolbachia in maintaining the quality of populations/lineages of C. flavipes.

Pebrine disease, caused by Nosema bombycis (Nb) infection in silkworms, is a severe and long-standing disease that threatens sericulture. As parasitic pathogens, a complex relationship exists between microsporidia and their hosts at the mitochondrial level. Previous studies have found that the translocator protein (TSPO) is involved in various biological functions, such as membrane potential regulation, mitochondrial autophagy, immune responses, calcium ion channel regulation, and cell apoptosis. In the present study, we found that TSPO expression in silkworms (BmTSPO) was upregulated following Nb infection, leading to an increase in cytoplasmic calcium, adenosine triphosphate, and reactive oxygen species levels. Knockdown and overexpression of BmTSPO resulted in the promotion and inhibition of Nb proliferation, respectively. We also demonstrated that the overexpression of BmTSPO promotes host cell apoptosis and significantly increases the expression of genes involved in the immune deficiency and Janus kinase-signal transducer and the activator of the transcription pathways. These findings suggest that BmTSPO activates the innate immune signalling pathway in silkworms to regulate Nb proliferation. Targeting TSPO represents a promising approach for the development of new treatments for microsporidian infections.

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is a highly destructive polyphagous pest that primarily damages maize. Maize is considered a most versatile crop for growing intercrops due to the wide row it needs. Maize–pea intercropping is preferred by small and marginal farmers worldwide due to various advantages including higher yield and improved economic benefits. However, the success of this intercropping system may be hampered if pea could sustain the FAW population. Thus, to clarify the fitness and potential effect of S. frugiperda on pea, we analysed the survival and development of S. frugiperda fed on pea leaves in the laboratory and constructed age-stage and two-sex life tables. Results showed that FAW successfully completed its life cycle when fed on pea and produced fertile offspring. The pre-adult duration was significantly higher on pea than maize. The net reproductive rate, intrinsic and finite rate of population increase on pea (135.06 offspring per individual, 0.12 offspring per individual per day and 1.13 times per day) were all significantly different from those on maize (417.64 offspring per individual, 0.19 offspring per individual per day and 1.21 times per day). The probability of survival of S. frugiperda at each stage was lower when fed on pea leaves than that of maize-fed larvae. Due to the overlapping growth periods of the maize and pea, S. frugiperda can easily proliferate throughout the year by shifting between adjacent crops. Thus, this study revealed the adaptability of S. frugiperda for pea and provides the foundation for further assessment of FAW risk to other inter-crops.

The social-sexual environment is well known for its influence on the survival of organisms by modulating their reproductive output. However, whether it affects survival indirectly through a variety of cues without physical contact and its influence relative to direct interaction remain largely unknown. In this study, we investigated both the indirect and direct influences of the social-sexual environment on the survival and reproduction of the mite Tyrophagus curvipenis (Acari: Acaridae). The results demonstrated no apparent influence of conspecific cues on the survival of mites, but the survival and reproduction of mated female mites significantly changed, with the females mated with males having a significantly shortened lifespan and increased lifetime fecundity. For males, no significant difference was observed across treatments in their survival and lifespan. These findings indicate that direct interaction with the opposite sex has a much more profound influence on mites than indirect interaction and highlight the urgent need to expand research on how conspecific cues modulate the performance of organisms with more species to clarify their impacts across taxa.

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.

Since its introduction in Spain in 2004, Aedes albopictus has rapidly spread across the country. Its aggressive biting behaviour causes nuisance, limiting outdoor activities. Also, its role as a vector of several arboviruses implies a major public health risk, with several cases of autochthonous dengue having been reported nationwide over the past few years. Control strategies usually focus on interventions in breeding sites. As such, accurate knowledge of the main larval habitats becomes a major priority in infested areas. A detailed identification of breeding sites of Ae. albopictus was carried out in the outdoors of 60 residential properties during July–August 2022 in El Vedat de Torrent (Valencia, Eastern Spain), an area recently colonised by this species. A total of 1444 real and potential breeding sites were examined. The most abundant potential larval habitat were plant pot plates (6.48 units/house), although a low infestation level was found, both for larvae (2.06% positivity, x̄ = 30.5 larvae/container), and pupae (0.51%, x̄ = 2.5 pupae/container). A total of 7715 larvae and 205 pupae were found in a disused flooded water pool depuration system. Animal drinkers, buckets and irrigation water containers were found to be the most common positive containers. No statistical difference was observed among the different container materials. A general statistical increase of 1 larva per 11.7 ml of water in breeding sites was detected. Breeding sites of other species such as Culex pipiens (n = 2) and Culex modestus (n = 1) were also rarely found in this residential area. To our knowledge, this is the first aedic index study carried out in Europe, and it provides valuable information about the main domestic breeding habitats of Ae. albopictus, which can greatly improve control programmes.

Dengue fever is a viral disease caused by one of four dengue stereotypes (Flavivirus: Flaviviridae) that are primarily transmitted by Aedes albopictus (Skuse) and Aedes aegypti (L.). To safeguard public health, it is crucial to conduct surveys that examine the factors favouring the presence of these species. Our study surveyed 42 councils across four towns within the Bhakkar district of Punjab Province, by inspecting man-made or natural habitats containing standing water. First, door-to-door surveillance teams from the district health department were assigned to each council to surveillance Aedes species and dengue cases. Second, data collection through surveillance efforts, and validation procedures were implemented, and the verified data was uploaded onto the Dengue Tracking System by Third Party Validation teams. Third, data were analysed to identify factors influencing dengue fever cases. The findings demonstrated the following: (1) Predominantly, instances were discerned among individuals who had a documented history of having travelled beyond the confines of the province. (2) Containers associated with evaporative air coolers and tyre shops were responsible for approximately 30% of the Aedes developmental sites. (4) Variability in temperature was responsible for approximately 45% of the observed differences in the quantity of recorded Aedes mosquito developmental sites. (5) Implementation of dengue prevention initiatives precipitated a 50% reduction in Aedes-positive containers, alongside a notable 70% decline in reported cases of dengue fever during the period spanning 2019 to 2020, while the majority of reported cases were of external origin. Aedes control measures substantially curtailed mosquito populations and lowered vector-virus interactions. Notably, local dengue transmission was eliminated through advanced and effective Aedes control efforts, emphasising the need for persistent surveillance and eradication of larval habitats in affected regions.

The Argentine ant (Linepithema humile) and the little fire ant (Wasmannia auropunctata) are among the top 100 invasive alien species globally, causing significant ecological and economic harm. Therefore, it is crucial to study their potential geographic distribution worldwide. This study aimed to predict their global distribution under current and future climate conditions. We used distribution data from various sources, including CABI, GBIF, and PIAKey, and key climate variables selected from 19 environmental factors to model their potential geographic distribution using MaxEnt. The AUC values were 0.925 and 0.937 for L. humile and W. auropunctata, respectively, indicating good predictive performance. Suitable areas for L. humile were mainly in southern North America, northern South America, Europe, central Asia, southern Oceania, and parts of Africa, while W. auropunctata suitable areas were mostly in southern North America, most of South America, a small part of Europe, southern Asia, central Africa, and some parts of Oceania. Under climate change scenario, suitable areas for L. humile increased, while highly suitable areas for W. auropunctata decreased. The top four countries with the largest areas of overlapping suitable habitat under current climate were Brazil, China, Australia, and Argentina, while under future SSP585 climate scenario, the top four countries were Brazil, China, Indonesia, and Argentina. Some countries, such as Estonia and Finland, will see an overlapping adaptation area under climate change. In conclusion, this study provides insight into controlling the spread and harm of L. humile and W. auropunctata.

Euwallacea interjectus, a recently discovered pest in poplar plantations, poses a significant economic threat due to its role in causing widespread tree mortality. This pest's cryptic behaviour has hindered research and control efforts, making laboratory rearing a valuable tool for studying its development and biology. We investigated the development period and biological characteristics of E. interjectus using artificial diets and fungal medium. Our findings revealed that the development time for eggs, larvae, and pupae averages approximately 6, 18, and 6 days, respectively. Notably, first and second instar larvae displayed peak moulting periods at 3.45 ± 0.64 SD and 7.92 ± 1.77 SD days, respectively. Furthermore, we measured head capsule widths of postmolt larvae, yielding values of 318.02 ± 7.38 SD μm for first-instar larvae, 403.01 ± 11.08 SD μm for second-instar larvae, and 549.54 ± 20.74 SD μm for third-instar larvae. Our research also uncovered a positive correlation between the number of progeny (eggs, larvae, pupae, and adults) and the mean length of the gallery system. Interestingly, the haplodiploid reproductive strategy did not significantly affect the number of offspring produced by the foundress. Additionally, we observed that foundresses displayed higher fecundity when subjected to nutrient-rich diets as compared to nutrient-poor diets. Our results will deepen our understanding of the biology of E. interjectus and provide criteria for larval instar classification. Additionally, managing nutrient availability within the colony could be considered a viable approach to regulating population size.

Aphidius colemani is an important biological control agent, used in greenhouses and open fields against aphid pests. Despite this economical importance, A. colemani, along with A. transcaspicus and A. platensis, has gone through a complex taxonomical history. The three species have only recently gained status as separate species again, comprising the morphologically defined Aphidius colemani species group. Other than sporadic records probably as a consequence of escape from greenhouses, the A. colemani species group members prefer warmer regions and there are numerous records from South America, Southern Europe and Asia. Based on slide-mounted material collected in the period 1964–2001 in Africa, we describe five new species belonging to this group, and report A. colemani, A. transcaspicus and A. platensis from several African countries. This data opens questions about the origin of the group and presents potential for the diversification of biological control agents against aphid pests.

The Culicidae mosquito family breeds in various natural and artificial water bodies. Phytotelmata are plant structures that hold water and are used by many mosquito species during their life cycle. Mosquitoes are disease vectors, and phytotelmata play an important role in harbouring them. Investigating these phytotelmata will facilitate to develop appropriate site specific vector control strategy. Approximately 750 phytotelmata in 16 locations on Car Nicobar Island was examined. Mosquito larvae were collected from different habitats such as tree holes, root holes, leaf axils, and bamboo stumps. On average, the number of larvae per 10 ml of water ranged from 0.003 to 3.833. Among the 16 mosquito species identified, there were vectors of malaria (Anopheles sundaicus), dengue (Aedes aegypti and Aedes albopictus) and filariasis (Culex quinquefasciatus), belonging to seven genera. The most common phytotelmata were coconut trees (17.5%) followed by Java apple trees (10.3%) and bamboo stumps (7.1%). The most widespread mosquito species was Aedes edwardsi, followed by Aedes malayensis and Ae. albopictus, and the per cent of phytotelmata that contained these species were 42.6, 22.0, and 16.8% respectively. Anopheles mosquitoes had the highest container index (6.1%) in root holes, while Aedes and Culex mosquitoes were more frequently found in bamboo stumps. The alpha biodiversity indices indicated that the mosquito population in Car Nicobar Island was highly diverse (S = 16; DMg = 36.9). Tree holes showed the highest diversity, as all 16 species were collected from them. Assessment of phytotelmata, there is a significant relationship observed between mosquito genus, village, habitat, and tree families, on the remote island of Car Nicobar. Considering the presence of disease-transmitting mosquito vectors in the phytotelmata, these habitats should be considered when implementing control measures to eliminate mosquito-borne infections.

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.

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.

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.

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.

Peatlands, shaped by centuries of human activities, now face a primary threat from mining activities. Vulnerable to drainage and hydrological instability, peatland areas encounter challenges that compromise their ecological integrity. This study hypothesised that permanent water reservoirs within mines could serve as refugia for water beetles from adjacent areas prone to drying in the summer. Employing standard methods, including entomological scraping and water traps, samples were collected. Results revealed that, in most cases, water beetles exhibited a preference for the Nature 2000 area untouched by mining. Despite unfavourable conditions, the Nature 2000 area showcased a more diverse water beetle fauna. Remarkably, the selected Nature 2000 area, despite its identified degradation based on flora, remained a biodiversity hotspot for peatland water beetle fauna. The study underscores the significance of assessing insects, particularly beetles, as rapid responders to environmental changes. This evaluation holds crucial implications for peatland restoration planning and decision-making regarding mining investments in proximity to peatland areas.

Lateralisation is a well-established phenomenon observed in an increasing number of insect species. This study aims to obtain basic details on lateralisation in courtship and mating behaviour in Ostrinia furnacalis, the Asian corn borer. We conducted laboratory investigations to observe lateralisation in courtship and mating behaviours in adult O. furnacalis. Our goal was also to detect lateralised mating behaviour variations during sexual interactions and to elucidate how these variances might influence the mating success of males. Our findings reveal two distinct lateralised traits: male approaches from the right or left side of the female and the direction of male turning displays. Specifically, males approaching females from their right side predominantly exhibited left-biased 180° turning displays, while males approaching females from the left-side primarily displayed right-biased 180° turning displays. Notably, left-biased males, executing a 180° turn for end-to-end genital contact, initiated copulation with fewer attempts and began copulation earlier than their right-biased approaches with left-biased 180° turning displays. Furthermore, mating success was higher when males subsequently approached the right side of females during sexual encounters. Left-biased 180° turning males exhibited a higher number of successful mating interactions. These observations provide the first report on lateralisation in the reproductive behaviour of O. furnacalis under controlled laboratory conditions and hold promise for establishing reliable benchmarks for assessing and monitoring the quality of mass-produced individuals in pest control efforts.

In internal parasitism, the respiration strategy within the host's body is as essential as evading attack from the host's immune system. Tachinid flies are parasitoids of terrestrial arthropods, mostly insects, during their larval stage. To obtain oxygen while living in the host body, they build a cylindrical structure known as the respiratory funnel at the aperture opened by the tachinid larva on the host integument or trachea. These funnels can be divided morphologically into sheath and cone types. Previous research on sheath-type funnels revealed that they are derived from the encapsulating substance produced by the host's immune system. In contrast, the cone-type funnels cover part of the body of the larval tachinid and may be constructed independently from the host immune system. To determine the mechanisms of cone-type funnel formation, histological observations were carried out on Gymnosoma rotundatum (L.) (Diptera: Tachinidae), which possesses this type of funnel. The respiratory funnel of G. rotundatum was found to be derived from the tube-shaped faeces wrapped with the peritrophic membrane and excreted by the fly larva, not from host tissue or haemocytes. Additionally, secretory glands putatively involved in the funnel formation were discovered around the larval anal plate of G. rotundatum. A comparison of funnel types within Tachinidae revealed that Phasiinae and Dexiinae have cone-type funnels, which may be created by the same mechanism as in G. rotundatum. These new findings suggest that funnel formation that does not use the host immune system is relevant to tachinid phylogeny.