Zinc finger protein (Zelda) of Tribolium castaneum (TcZelda) has been showed to play pivotal roles in embryonic development and metamorphosis. However, the regulatory mechanism of TcZelda associated with these physiology processes is unclear. Herein, the developmental expression profile showed that Zelda of T. castaneum was highly expressed in early eggs. Tissue expression profiling revealed that TcZelda was mainly expressed in the larval head and adult ovary of late adults and late larvae. TcZelda knockdown led to a 95% mortality rate in adults. These results suggested that TcZelda is related to the activation of the zygote genome in early embryonic development. Furthermore, 592 differentially expressed genes were identified from the dsZelda treated group. Compared with the control group, altered disjunction (ALD) and AGAP005368-PA (GAP) in the dsZelda group were significantly down-regulated, while TGF-beta, propeptide (TGF) was significantly up-regulated, suggesting that TcZelda may be involved in insect embryonic development. In addition, the expression of Ubx ultrabithorax (UBX), Cx cephalothorax (CX), En engrailed (EN), and two Endocuticle structural glycoprotein sgabd (ABD) genes were significantly down-regulated, suggesting that they may cooperate with TcZelda to regulate the development of insect wings. Additionally, Elongation (ELO), fatty acid synthase (FAS), and fatty acyl-CoA desaturase (FAD) expression was inhibited in dsZelda insects, which could disturb the lipase signaling pathways, thus, disrupting the insect reproductive system and pheromone synthesis. These results may help reveal the function of TcZelda in insects and the role of certain genes in the gene regulatory network and provide new ideas for the prevention and control of T. castaneum.

Eupeodes corollae (F.) (Diptera: Syrphidae) is the most abundant syrphid fly which is distributed worldwide and is the sole predator of aphids. Therefore, the present study was conducted to evaluate the predation rate and functional response of E. corollae against the cabbage aphid, Brevicoryne brassicae (L.). The experiment was carried out under laboratory conditions at 25 ± 2°C with 60–70% relative humidity. The results revealed that age-specific net predation rate (qx) increased after the 4th day and a peak was recorded on the 10th day of pivotal age in the third larval instar. The stable host kill rate and finite host kill rate of E. corollae were 18.63 and 21.07, respectively, against the B. brassicae and predicted that a mean of 20.78 aphids was needed for E. corollae to produce one offspring. A negative linear coefficient (P < 0) indicated the type II functional response for all larval instars of E. corollae against the B. brassicae. At higher prey density, the prey consumption was significantly at par with second and third instar larvae of E. corollae as the prey consumption was increased with increasing the prey density, which then decreased after attaining the upper asymptote (76.40 and 81.40% consumption, respectively). The Roger's predator random equation for type II functional response was fitted to estimate attack rate (a) and handling time (Th). The maximum prey consumption was recorded for third instar of E. corollae with a higher attack rate (0.336 h−1) and lower handling time (0.514 h) against B. brassicae, followed by the second and first instar. Thus, it is concluded that the third larval instar of E. corollae was the voracious feeder and used as an efficient biocontrol agent in the IPM programme.

Parasitoids are an important group of insects because their species number is among the highest. Multiple studies have addressed the relationships between forest successional age and insect diversity by focusing on herbivorous organisms, but changes in diversity of parasitoids are still poorly known. This work analyses the diversity of parasitoids in tropical forests representing three successional stages. A total of 30 traps were placed, ten in each forest successional stages. We estimated true diversity of Ichneumonidae species and guilds and explored the relationship between their diversity and the abundance of plant species using an Indicator Species Analysis; the relationship between parasitoid species and plant richness and abundance was tested using a Redundancy Analysis. A total of 1522 individuals and 168 morpho-species were captured in four months. Species richness showed no differences; however, parasitoid abundance was higher in young forest, while intermediate forest had the highest true diversity values (1D) with 71.6 effective species. According to insect guilds, richness, abundance, and diversity were similar in the three vegetation successional stages. This finding may be explained based on the intermediate disturbance hypothesis, which postulates that moderate disturbance levels favor the highest diversity. In conclusion, successional age matters, i.e., diversity is the highest in intermediate stages, while the old forests harbors guilds unique to that successional stage, such as parasitoids of melitophagous larvae of bees. Other successional stages were characterized by a single species of parasitoid, belonging to the genera Eiphosoma and Anomalon, which may indicate altered and preserved forests, respectively.

Rhynchophorus ferrugineus is a quarantine pest that mainly damages plants in tropical regions, which are essential economic resources. Cry3Aa has been used to control coleopteran pests and is known to be toxic to R. ferrugineus. The binding of the Cry toxin to specific receptors on the target insect plays a crucial role in the toxicological mechanism of Cry toxins. However, in the case of R. ferrugineus, the nature and identity of the receptor proteins involved remain unknown. In the present study, pull-down assays and mass spectrometry were used to identify two proteins of aminopeptidase N proteins (RfAPN2a and RfAPN2b) in the larval midguts of R. ferrugineus. Cry3Aa was able to bind to RfAPN2a (Kd = 108.5 nM) and RfAPN2b (Kd = 68.2 nM), as well as midgut brush border membrane vesicles (Kd = 482.5 nM). In silico analysis of both RfAPN proteins included the signal peptide and anchored sites for glycosyl phosphatidyl inositol. In addition, RfAPN2a and RfAPN2b were expressed in the human embryonic kidney 293T cell line, and cytotoxicity assays showed that the transgenic cells were not susceptible to activated Cry3Aa. Our results show that RfAPN2a and RfAPN2b are Cry3Aa-binding proteins involved in the Cry3Aa toxicity of R. ferrugineus. This study deepens our understanding of the action mechanism of Cry3Aa in R. ferrugineus larvae.

Phenoloxidase (PO) is a significant biomolecule involved in humoral defence mechanism of invertebrates. Spontaneous melanization of insect haemolymph is the major hinderance for studying PO activity, as haemolymph was collected devoid of phenylthiourea. In the study, no visible melanization was observed in crude serum from the grub of Oryctes rhinoceros up to 30 min of incubation amongst crude haemolymph, diluted haemolymph, crude serum and diluted serum that were subjected to visual observation for spontaneous melanization reaction. Accordingly, crude serum was taken for evaluating PO activity. At the same time, as PO substrates tend to auto-oxidize and provide false optical density value, tris-buffered saline devoid of any substrates were used as blank for PO assays. The ideal wavelength at which maximum PO activity occurred for each substrate, namely, tyrosine, tyramine, dopamine, L-dopa, DL-dopa, catechol, protocatechuic acid and pyrogallol was determined as 407, 410, 429, 465, 403, 466, 428 and 400 nm, respectively. Additionally, time course of oxidation for each phenolic substrate by the serum PO were examined and DL-dopa was identified as the specific substrate for serum PO in the grub of O. rhinoceros. Furthermore, maximum PO activity was observed at 5 min of incubation for 10 mM of DL-dopa that was considered as optimum concentration. The ideal pH and temperature for serum PO activity was observed as 7.5 and 20°C, respectively. These results suggested that standardizing a suitable substrate is an essential prerequisite to evaluate the real PO activity of serum which might significantly fluctuate in each insect model.

Spotted lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae), is an invasive insect that was first detected in the United States in 2014 and feeds on a wide variety of plants, with economic impacts on the agricultural, ornamental, and timber industries. Part of what likely contributes to the success of L. delicatula in its invaded range is that it appears to be chemically defended by sequestering toxins from its host plant(s), which may deter predators in the introduced range. To determine the identity and behavior of North American predators that feed on spotted lanternfly, we performed a community science study in which we asked members of the public to contribute reports of animals feeding on spotted lanternfly through a Facebook page. The largest group of reported predators was arthropods followed by birds. Araneae was the arthropod order with the most reports and Phasianidae was the most frequently reported bird family. Using Pearson's χ2 tests, we also identified significant relationships between predator behavior and (1) taxonomic group of the predator, (2) L. delicatula life stage, and (3) host plant L. delicatula was observed on. These results can help to guide future research on predator host shifting to spotted lanternfly and potential for biocontrol as a management tactic.

Galls function as provide shelter for gall inducers, guarding them against their natural enemies. Previous research has illuminated the interactions between galls, gall inducers, and their corresponding parasitoids within various caltrop plants. However, less is known about these relationships within Nitraria sibirica, particularly regarding the efficacy of parasitism. Therefore, this study aimed to identify the morphometric relationships among the swollen galls, gall inducers, and their parasitoids. Two species of gall inducers and three species of parasitoids were obtained from the swollen galls of N. sibirica. The correlations of the parasitization indexes, the lifespan of gall inhabitants, and temperature and the morphometric relationships between the galls and their inhabitants were analyzed. The dominant gall inducer identified was Contarinia sp. (Diptera: Cecidomyiidae). Furthermore, it was observed that three solitary parasitoids attacked Contarinia sp. in the swollen galls, with only Eupelmus gelechiphagus acting as an idiobiont ectoparasitoid. The dominant parasitoids were Platygaster sp. and Cheiloneurus elegans at sites 1 and 2, respectively, with Platygaster sp. displaying greater abundance than C. elegans in the swollen galls. The lifespan of the gall inhabitants shortened gradually as the temperature increased. Moreover, the optimal number of gall chambers ranged from two to four per swollen gall with maximized fitness, which can be considered the optimal population density for the gall inducer Contarinia sp. Morphometric analysis exhibited a strong linear correlation between gall size and chamber number or the number of gall inhabitants, as well as a weak correlation between gall size and body size of the primary inhabitants of swollen galls. Our results highlight the importance of the biological investigation of parasitoids and gall inducers living in closed galls with multiple chambers and may pave the way for potential application in biological control.

Aphids exhibit seasonally alternating asexual and sexual reproductive modes. Different morphs are produced throughout the life cycle. To evaluate morph-specific fitness during reproductive switching, holocyclic Sitobion avenae were induced continuously under short light conditions, and development and reproduction were compared in each morph. Seven morphs, including apterous and alate virginoparae, apterous and alate sexuparae, oviparae, males, and fundatrices, were produced during the life cycle. The greatest proportions of sexuparae, oviparae, males, and virginoparae were in the G1, G2, G3, and G4 generations, respectively. Regardless of asexual or sexual morphs, alate morphs exhibited a marked delay in age at maturity compared with that of apterous morphs. Among the alate morphs, males had the longest age at maturity, followed by sexuparae and virginoparae. Among the apterous morphs, sexuparae were older at maturity than the fundatrices, virginoparae, and oviparae. The nymphs of each morph had equal survival potentials. For the same wing morphs, apterous sexuparae and oviparae exhibited substantial delays in the pre-reproductive period and considerable reductions in fecundity, compared with those of apterous virginoparae and fundatrices, whereas alate sexuparae and alate virginoparae had similar fecundity. The seven morphs exhibited Deevey I survivorship throughout the life cycle. These results suggest that sexual production, particularly in males, has short-term development and reproduction costs. The coexistence of sexual and asexual morphs in sexuparae offspring may be regarded as an adaptive strategy for limiting the risk of low fitness in winter.

Environment-induced epigenetics are involved in diapause regulation, but the molecular mechanism that epigenetically couples nutrient metabolism to diapause regulation remains unclear. In this study, we paid special attention to the significant differences in the level of N6-adenosine methylation (m6A) of dihydroxyacetone phosphate acyltransferase (DHAPAT) and phosphatidate phosphatase (PAP) genes in the lipid metabolism pathway of the bivoltine silkworm (Bombyx mori) strain Qiufeng developed from eggs incubated at a normal temperature (QFHT, diapause egg producer) compared to those from eggs incubated at a low temperature (QFLT, non-diapause egg producer). We knocked down DHAPAT in the pupal stage of the QFLT group, resulting in the non-diapause destined eggs becoming diapausing eggs. In the PAP knockdown group, the colour of the non-diapause destined eggs changed from light yellow to pink 3 days after oviposition, but they hatched as normal. Moreover, we validated that YTHDF3 binds to m6A-modified DHAPAT and PAP mRNAs to promote their stability and translation. These results suggest that RNA m6A methylation participates in the diapause regulation of silkworm by changing the expression levels of DHAPAT and PAP and reveal that m6A epigenetic modification can be combined with a lipid metabolism signal pathway to participate in the regulation of insect diapause traits, which provides a clearer image for exploring the physiological basis of insect diapause.

Chemosensory proteins (CSPs) were necessary for insect sensory system to perform important processes such as feeding, mating, spawning, and avoiding natural enemies. However, their functions in non-olfactory organs have been poorly studied. To clarify the function of CSPs in the development of Mythimna separata (Walker) larvae, two CSP genes, MsCSP17 and MsCSP18, were identified from larval integument transcriptome dataset. Both of MsCSP17 and MsCSP18 contained four conserved cysteine sites (C × (6)-C × (18)-C × (2)-C), with a signal peptide at the N-terminal. RT-qPCR analysis showed that MsCSP17 and MsCSP18 have different expression patterns among different developmental stages and tissues. MsCSP17 was highly expressed in 1st–4th instar larvae, and MsCSP18 had high expression in adults. Both genes were expressed highly in larval head, thorax, integument and mandible. Moreover, both of MsCSP17 and MsCSP18 were lowly expressed in larval integuments when larvae molted for 6 h and 9 h from 3rd to 4th instar, but highly at the beginning and end phase during molting. After injection of dsMsCSP17 and dsMsCSP18, the expression levels of two genes decreased significantly, with the body weight of larvae decreased, the mortality increased, and the eclosion rate decreased. It was suggested that MsCSP17 and MsCSP18 contributed to the development of M. separata larvae.

Major efforts to control the population of Aedes aegypti mosquitoes involve the use of synthetic insecticides, which can be harmful to the environment. Most plant compounds are eco-friendly and some of them have biocontrol potential, whereas a fraction of these compounds is released into the environment through the leaf-leaching process. We evaluated the effects of secondary compounds from Ateleia glazioviana and Eucalyptus grandis senescent leaf leachates on Ae. aegypti larval mortality, adult emergence time, and wing size using a microcosm approach. The microcosms consisted of 10 larvae kept in water (control) and under four treatments with leachates from a combination of plant species and leaching time (7 or 14 days). Chemical analyses of the leachates showed the presence of carboxaldehyde and Heptatriocotanol, which have antimicrobial properties, potentially reducing the food available for larvae. β-Sitosterol, Stigmasterol, α-Amyrin, and Lupeol are compounds with inhibitory, neurotoxic, and larvicidal effects. Both plant species’ leachates increased larval mortality and decreased emergence time due to the presence of compounds toxic to the larvae. Larger organisms emerged in treatments with 7-days leachates, likely due to the high concentration of dissolved organic matter in the leachates. The higher mortality in 7-days leachates may also increase the organic matter from co-specific decomposition, improving adult size. Therefore, if the mosquito population is not locally extinct, compounds present in leaf leachates may act as a resource enhancing larvae growth, potentially increasing survivors’ fitness. In conclusion, biocontrol attempts using urban green spaces may have unexpected outcomes, such as resulting in larger pest organisms.

The honey bee is associated with a diverse community of microbes (viruses, bacteria, fungi, and protists), commonly known as the microbiome. Here, we present data on honey bee microbiota from two localities having different surrounding landscapes – mountain (the Rhodope Mountains) and lowland (the Danube plain). The bacterial communities of abdomen of adult bees were studied using amplicon sequencing of the 16S rRNA gene. The composition and dominance structure and their variability within and between localities, alpha and beta diversity, and core and differential taxa were compared at different hierarchical levels (operational taxonomic units to phylum). Seven genera (Lactobacillus, Gilliamella, Bifidobacterium, Commensalibacter, Bartonella, Snodgrassella, and Frischella), known to include core gut-associated phylotypes or species clusters, dominated (92–100%) the bacterial assemblages. Significant variations were found in taxa distribution across both geographical regions and within each apiary. Lactobacillus (Firmicutes) prevailed significantly in the mountain locality followed by Gilliamella and Bartonella (Proteobacteria). Bacteria of four genera, core (Bartonella and Lactobacillus) and non-core (Pseudomonas and Morganella), dominated the bee-associated assemblages of the Danube plain locality. Several ubiquitous bacterial genera (e.g., Klebsiella, Serratia, and Providencia), some species known also as potential and opportunistic bee pathogens, had been found in the lowland locality. Beta diversity analyses confirmed the observed differences in the bacterial communities from both localities. The occurrence of non-core taxa contributes substantially to higher microbial richness and diversity in bees from the Danube plain locality. We assume that the observed differences in the microbiota of honey bees from both apiaries are due to a combination of factors specific for each region. The surrounding landscape features of both localities and related vegetation, anthropogenic impact and land use intensity, the beekeeping management practices, and bee health status might all contribute to observed differences in bee microbiota traits.

DNA Barcoding is an important tool for disciplines such as taxonomy, phylogenetics and phylogeography, with Barcode of Life Data System (BOLD) being the largest database of partial cytochrome c oxidase subunit I (COI) sequences. We provide the first extensive revision of the information available in this database for the insect order Thysanoptera, to assess: how many COI sequences are available; how representative these sequences are for the order; and the current potential of BOLD as a reference library for specimen identification and species delimitation. The COI database at BOLD currently represents only about 5% of the over 6400 valid thrips species, with a heavy bias towards a few species of economic importance. Clear Barcode gaps were observed for 24 out of 33 genera evaluated, but many outliers were also observed. We suggest that the COI sequences available in BOLD as a reference would not allow for accurate identifications in about 30% of Thysanoptera species in this database, which rises to 40% of taxa within Thripidae, the most sampled family within the order. Thus, we call for caution and a critical evaluation in using BOLD as a reference library for thrips Barcodes, and future efforts should focus on improving the data quality of this database.

Termite colony foundation precedes the incipient stage, when the first oviposition cycle takes place, followed by months of reproductive inactivity. The royal couple is supposed to cease oviposition during this period, investing energy to care for the first brood. When a suitable number of alloparents differentiate, egg-laying resumes. Here we followed oviposition dynamics, embryo development and queen/king body changes in laboratory colonies of the major pest species Coptotermes gestroi (Rhinotermitidae) and Cryptotermes brevis (Kalotermitidae) during 9 months. We show that they differ in these oviposition dynamics, as C. gestroi queens displayed an uninterrupted oviposition whereas C. brevis laid a cohort of eggs and ceased oviposition during a 3-month period (lag phase). C. gestroi oviposition dynamic was remarkable and suggests that occurrence of progeny was not a limiting factor, thus queens and kings were able to concomitantly invest energy in reproduction and parental care. These findings contrast those reported for rhinotermitids from temperate areas, and we discuss the likely reasons for such a condition, including endogenous rhythms, avoidance of a high mortality rate of the first progeny and adaptation to the weather conditions of the Neotropical region. Oviposition dynamic in C. brevis resembled those of several termite species, in which the royal couple cease reproduction to care for the first brood. Rearing conditions did not influence oviposition dynamics (egg-laying cycle followed by a lag phase), thus our results on the oviposition of C. gestroi and C. brevis correspond to different reproductive strategies post-foundation adopted by these pest species.