Search

The stability of symbiotic flora is an important indicator of the health of an organism. Symbiotic bacteria have been proven to be closely involved in the immune process of organisms. The pathogenicity of Beauveria bassiana was studied in relation to symbiotic bacteria on the surface and inside of the migratory locust (Locusta migratoria). The results showed that the surface disinfection of test locusts contributed to the pathogenicity of B. bassiana to locusts. Most of the surface bacteria of L. migratoria caused some inhibition of B. bassiana growth, and LM5-4 (Raoultella ornithinolytica), LM5-2 (Enterobacter aerogenes), and LM5-13 (Citrobacter freundii) showed the highest inhibitory effect on the growth of B. bassiana. The inoculation of locusts with additional surface symbiotic bacteria reduced the virulence of B. bassiana to L. migratoria. Infection by different strains of B. bassiana caused similar changes in the symbiotic flora of migratory locusts. The inoculation of locusts with additional intestinal symbiotic bacteria (Enterobacter sp.) reduced the virulence of B. bassiana to L. migratoria. These findings illustrate the effect of bacterial communities on fungal infections in L. migratoria when seen from the perspective of ecology in a microenvironment. The active antifungal substances of such bacteria and their mechanisms of action need further study.

Body size is influenced by various environmental and genetic factors. The migratory locust, Locusta migratoria L., exhibits phase polyphenism in which nymphs change body coloration and adult body dimensions undergo specific changes in response to different population densities experienced during nymphal development. In this study, I collected adults of the migratory locust L. migratoria in solitarious (low-density) populations in central Japan over 16 years to determine how adult body size changed with years and how stable their body dimensions were. Four size parameters including head width (C), hind femur length (F), forewing length (E) and total body length (TBL) were highly correlated with each other. None of these parameters correlated with years except for E with a marginally significant correlation. None of these parameters showed a significant correlation with mean annual temperatures. The classical morphometric ratios, F/C and E/F, which are often used to separate different phases, showed no significant differences among years, suggesting that they may serve as reliable phase indicators for solitarious populations. The sexual size dimorphism (SSD) was confirmed and consistently larger variances in adult body size were observed in females than in males. When the mean values of the two sexes in different years were plotted, no significant correlation was observed in any of the body size parameters including C, F and E, indicating that the two sexes varied these parameters independently. The present data may provide a basis to analyze body size changes in relation to global climate changes in the future.

Locust is the main pest endangering Chinese agriculture and even global agriculture. Therefore, the development of an efficient and real-time locust monitoring technology is of great significance to the prevention and management of locust disasters. Video monitoring technology has been widely used in locust monitoring, but the different postures of locusts in the field and the problem of plant shelter pose great challenges to accurate locust monitoring. In this paper, taking a 5-year-old migratory locust as the research object, video target detection based on the MOG2-YOLOv4 network is proposed to address the problem of video frame recognition failure caused by different postures and partial occlusion during locust movement. First, the frame compensation algorithm is used to identify locusts with different postures; then the background separation algorithm (MOG2) is used to extract the spatio-temporal features of the next and previous frames of the video, and the combination of the MOG2 algorithm and YOLOv4 algorithm to extract the static features of locusts in the video can improve the detection accuracy. Experimental results show that the average detection accuracy of the MOG2-YOLOv4 model is 82.33% (8.17% higher than that of YOLOv4), and the F-mean is 89.37% (4.76% higher). Therefore, this method has a good effect on the identification of East Asian migratory locusts, particularly under occlusion and motion blur conditions; it achieves accurate monitoring of the occurrence of locust plagues, which is of great significance to the prevention and control of locust plagues.

Short horned orthopetra from the family acrididae in gregarious phase is generally known as locust. In solitary condition they are harmless and overlooked. In history no epidemic of locust was snoop in the cold desert of Ladakh. In the last two decades Locusta migratoria migratoria (Linnaeus, 1758) is enormously affecting the livelihood of the trans-Himalayan region. Present study emphasized in Changthang, Chusul and Pangom Tso area of the Ladakh from 2008 to 2019. The purpose of this study was to explore the distribution, ecological and taxonomical aspects of the outbreak of migratory locusts in the fragile ecosystem. The long term monitoring assessed the geographic coverage of the locust infestation, swarming hotspots; evaluate the population parameters such as densities and developmental stages in the region. It was observed that migratory locust impinge along the Indus river area from Tibet and China and swarms year to year in Ladakh. They drastically impacted on the livelihood of the region with demolish the greenery of the area.

Cuticular proteins (CPs) are major components of the insect cuticle-associated organs such as integument and wings, although the importance of CPs for wing development and function in hemimetabolous insects remains understudied. In the present study, a wing cuticular protein LmACP8 was identified from Locusta migratoria, which belongs to the RR-2 subfamily of cuticular protein R&R consensus (CPR) chitin-binding proteins. LmACP8 was mainly expressed in the wing pads and showed high expression levels before ecdysis of third-, fourth-, and fifth-instar nymphs, with its encoded protein located in the procuticle of wing pads and adult wings. Depletion of LmACP8 by RNA interference markedly reduced the amount of its protein, which consequently caused abnormal wing morphogenesis in the transition from nymph to adult of L. migratoria. We further demonstrated that the abnormal morphogenesis was caused by severe damage of the endocuticle in the wings. LmACP8 was suppressed by 20-hydroxyecdysone (20E) in vivo, however, its expression was significantly up-regulated after knocking down the hormone receptor gene LmHR39. Thus, the LmACP8 that is negatively regulated by the LmHR39-mediated 20E signaling pathway is involved in wing development during the nymph to adult transition.

Evolutionary immune system upgradation is done by all living organism to survive from the invading pathogens. Insects has a strong defense system comprising of anatomical barrier, humoral antimicrobial peptide (AMP) production and cellular immune components. Locusta migratoria (L. migratoria) is an agricultural pest insect migrating long distance might encounter distinct pathogens. Owing to this, we aimed in identifying AMPs present in the L. migratoria and elucidate the antimicrobial activity. The migratory locust was immunized with Escherichia coli, Staphylococcus aureus, Candida albicans mixture, then RNA was isolated and sequenced. The L. migratoria transcriptome was de novo assembled using trinity and screened for AMPs propensity specifically molecular and aggregation properties. The novelty is tested by blasting in known AMP databases. Finally, obtained novel putative AMPs were then tested for antimicrobial and hemolytic activity. The prediction resulted in 3,524 putative AMPs which was further screened down to ten AMPs for final testing. Two peptides showing promising antimicrobial effects were obtained. All the analyzed peptides showed no hemolysis confirms its non-toxicity. Thus, our peptides could be promising drug candidate and can be used as an alternative to antibacterial or antifungal therapy with further validations.

PurposeThe effects of incorporation of Locusta migratoria (LM) powder at different levels (0, 1, 2, 3, 4 and 5%) on nutritional, qualitative and sensory properties of baked breads were investigated.Design/methodology/approachChemical composition, mineral elements and functional properties of wheat flour, LM powder and their binary mixture systems were determined. The breads were assayed for proximate composition, minerals, amino acids profile, physical characteristics and sensorial properties.FindingsLM powder have relatively high levels of protein (51.97% db), fat (23.15% db) and fibers (13.76% db). LM powder presented significantly (p ≤ 0.05) greater water absorption capacity (WAC), oil absorption capacity(OAC), emulsion activity (EA) and foam capacity (FC) as compared to wheat flour. Blending wheat flour with various levels of LM powder significantly (p ≤ 0.05) improved the functional properties of binary mixture systems. Protein, fat, fiber and ash contents of bread samples significantly increased with the addition of LM powder. No significant differences (p ≤ 0.05) were recorded in specific volume between control breads and those breads containing LM powder up to 2%. Breads fortified with LM powder contained higher values of essential amino acids than the limits of FAO/WHO pattern, with exception of lysine. Sensory evaluation results showed that bread samples with the addition of LM powder at levels 1–4% had high overall acceptability.Research limitations/implicationsLimitations of our study are the lack of approved and professional programs about management, collection, harvesting, processing, marketing and consumption of edible insects; therefore, these results should be generalized implications for industry. This present study, therefore, provides useful data to support public health nutrition aimed at improving the nutritional health of populations through the promotion of bakery products enriched with LM powder.Practical implicationsThe main findings of this study indicated that the addition of 1–4% of LM powder into wheat flour enhanced sensory and nutritional properties of produced breads.Originality/valueIt is concluded that the addition of LM powder improves the nutritional and sensorial properties of the produced bread when the wheat flour is substituted with 1–4% of LM powder.

The desert locust, Schistocerca gregaria (Forsk.) (Orthoptera: Acrididae), avoids laying egg pods in sand containing locust fecal extracts. In this study, the oviposition inhibitory (OI) effect was investigated for a different locust species, the migratory locust Locusta migratoria (L.) (Orthoptera: Acrididae). Females of L. migratoria laid more egg pods in control sand than in sand containing fecal extracts obtained from L. migratoria and S. gregaria. However, L. migratoria oviposition was less sensitive to fecal extracts than was S. gregaria oviposition. The OI effect was observed in L. migratoria irrespective of whether the fecal extracts were prepared from gregarious or solitarious locusts, suggesting that phase-specific metabolism is not involved in OI activity. Heating and bleaching of the fecal extracts reduced the amounts of fungi and bacteria in the extracts; however, the resultant extracts still led to OI behavior in L. migratoria. These results suggest that heat-stable and bleach-stable factors in fecal extracts are responsible for the OI effect in L. migratoria.

Reproductive synchrony generally occurs in various group-living animals. However, the underlying mechanisms remain largely unexplored. The migratory locust,Since 2019, a plague of flying insects known as migratory locusts has been causing extensive damage to crops in East Africa. Migratory locusts sometimes live a solitary lifestyle but, if environmental conditions allow, they form large groups containing millions of individuals known as swarms that are responsible for causing locust plagues.Locusts are able to maintain such large swarms because they can aggregate and synchronize. When they live in swarms, individual locusts produce odors that are sensed by other individuals in the group. For example, an aggregation pheromone, called 4-vinylanisole, is known to help keep large groups of locusts together. However, it is less clear how odors synchronize the reproductive cycles of the females in a swarm so that they are ready to mate with males and lay their eggs at the same time. To address this question, Chen et al. examined when female locusts reached sexual maturity after they were exposed to odors produced by other locusts living alone or in groups. The experiments found that only 4-vinylanisole, which was abundantly released by adult male locusts living in groups, stimulated female locusts to reach sexual maturity at the same time. This odor increased the levels of a hormone known as juvenile hormone in less-developed females to help them reach sexual maturity sooner. These findings demonstrate that when migratory locusts are living in swarms, male locusts promote the female locusts to reach sexual maturity at the same time by promoting less-developed females to ‘catch up’ with other females in the group. A next step will be to investigate the neural and molecular mechanisms underlying the ‘catch up’ effect induced by 4-vinylanisole.

Locust plagues threaten agricultural and environmental safety throughout the world1,2. Aggregation pheromones have a crucial role in the transition of locusts from a solitary form to the devastating gregarious form and the formation of large-scale swarms3,4. However, none of the candidate compounds reported5–7 meet all the criteria for a locust aggregation pheromone. Here, using behavioural assays, electrophysiological recording, olfactory receptor characterization and field experiments, we demonstrate that 4-vinylanisole (4VA) (also known as 4-methoxystyrene) is an aggregation pheromone of the migratory locust (Locusta migratoria). Both gregarious and solitary locusts are strongly attracted to 4VA, regardless of age and sex. Although it is emitted specifically by gregarious locusts, 4VA production can be triggered by aggregation of four to five solitary locusts. It elicits responses specifically from basiconic sensilla on locust antennae. We also identified OR35 as a specific olfactory receptor of 4VA. Knockout of OR35 using CRISPR–Cas9 markedly reduced the electrophysiological responses of the antennae and impaired 4VA behavioural attractiveness. Finally, field trapping experiments verified the attractiveness of 4VA to experimental and wild populations. These findings identify a locust aggregation pheromone and provide insights for the development of novel control strategies for locusts. 4-Vinylanisole, which is emitted by gregarious locusts or as a result of aggregation of solitary locusts, is identified as an aggregation pheromone that strongly attracts both solitary and gregarious locusts, acting via the olfactory receptor OR35.

BACKGROUND Knickkopf (Knk) proteins play crucial roles in the formation of insect cuticle. Recent studies in the holometabolous insect red flour beetle (Tribolium castaneum) have shown that three Knk genes encoding TcKnk, TcKnk2 and TcKnk3 play different but essential roles at different developmental stages and in different tissues. However, the functions of Knk genes had not been fully explored in hemimetabolous insects such as the migratory locust Locusta migratoria. RESULTS We identified three transcripts of LmKnk-like genes LmKnk2 and LmKnk3 with the full-length cDNA sequences, which were named as LmKnk2, LmKnk3-FL and LmKnk3-5'. These three transcripts were highly expressed before molting and mainly expressed in the integument. Among these genes, silencing only LmKnk3-5' by RNA interference (RNAi) caused molting defects and high mortality of the locusts. Injection of dsLmKnk3-5' dramatically decreased chitin content, but did not affect cuticle laminar ultra-structures in the integument. After the knockdown of LmKnk3-5' transcript, lipid deposition on the cuticle surface was impeded, and locusts exhibited increased susceptibility to each of four insecticides in three different classes. However, no visible phenotypic changes were observed after LmKnk2 or LmKnk3-FL was silenced by RNAi. CONCLUSION We demonstrate that LmKnk3-5' is essential for cuticle formation in L. migratoria. This contrasts the findings that the cognate protein in T. castaneum TcKnk3-5' is dispensable for cuticle formation and survival. Hence, we provide some evidence that the function of Knk-type proteins may be species-specific. We therefore think that LmKnk3-5' may be a good target for the application of RNAi-based technologies for species-specific insect pest management. © 2020 Society of Chemical Industry.

Diapause is a state of arrested growth, which allows insects to adapt to diverse environments. Serine protease inhibitors (serpins) play an important role in various physiological processes, including blood coagulation, fibrinolysis, development, complement activation and extracellular matrix remodeling. We hypothesized that serpin may affect energy metabolism and thereby control diapause of migratory locust (Locusta migratoria) embryos by regulating protease cascades. A total of seven nonredundant serpin genes (named serpin1–serpin7) of L. migratoria were obtained through transcriptomic analysis. We further performed label‐free proteomic sequencing and analysis of diapause and nondiapause eggs of L. migratoria, revealing significant differences in serpin7 expression. A significant reduction in diapause rate under the short photoperiod was observed in insects treated with serpin7 double‐stranded RNA. Furthermore, knockdown of the serpin7 gene resulted in significant upregulation of the activity of polyphenol oxidase. We therefore propose that the observed serpin7 gene plays a crucial role in diapause, suggesting that control of energy metabolism may have potential as a future strategy for the reproductive control of insect pests., Lm_Serpin7, a member of the Serpin protease inhibitor family, is highly expressed in diapause eggs and plays an important role in Locusta migratoria egg diapause. Knockdown of Lm_Serpin7 by injection with ds_serpin7 decreases egg diapause rate, suggesting that control of Lm_Serpin7 may have potential as a future strategy for the reproductive control of insect pests.

FMRFamide-related peptides (FaRPs) are a type of neuropeptide, which participate in a variety of physiological processes in insects. Previous study showed that myosuppressin, being a member of FaRPs, initiated pupal diapause in Mamestra brassicae. We presumed that FaRPs genes might play a critical role in photoperiodic diapause induction of L. migratoria. To verify our hypothesis, flrf, a precursor gene of FaRP from L. migratoria, was initially cloned under long and short photoperiods that encoded by flrf gene identified from central nervous system (CNS). Phylogenetic analysis showed that the protein encoded by L. migratoria flrf gene, clustered together with Nilaparvata lugens (Hemiptera: Delphacidae) with 100% bootstrap support, was basically an FMRFamide precursor homologue. We noticed the availability of -RFamide peptides (GSERNFLRFa, DRNFIRFa) under short photoperiod only, which suggested their functions related to photoperiodic diapause induction. RNAi and quantitative real-time PCR (qRT-PCR) results further confirmed that the flrf gene promoted locust's diapause.

The outbreak of Oriental Migratory Locust(Locusta migratoria manilensis) causes devastating disasters to agriculture. With the impact of climate changes and human activities, the distribution of locust habitat (locust habitat is the environment in which locusts live and survive) in China is constantly changing. Monitoring and extracting locust habitat are of great significance for guiding large-scale agricultural production. The occurrence of the locust is closely related to their habitat. Therefore, a comprehensive analysis of habitat factors that affect locust survival is carried out to monitor locust habitat distribution. Besides, the landscape structure also affects distribution. This study explored a model for analyzing multi-temporal Landsat and MODIS images, which combined multiple habitat factors and landscape structure to analyze locust habitat. The locust habitat near North Dagang Reservoir in Tianjin is the research object. First, the habitat factors that affect locust oviposition and growth were analyzed, and vegetation coverage, land cover class, soil moisture, soil salinity, and land surface temperature were selected as five habitat factors. The weights of five habitat factors were evaluated according to the Analytic Hierarchy Process (AHP) model. Then, considering the impact of landscape structure on locust habitat, a moving-window was used to correlate locust habitat factors at pixel scale with locust habitat at patch scale. Finally, the distribution map of the locust habitat at patch scale was generated. The Analytic Hierarchy Process(AHP) was used to compare and test the results. Our research shows that the Patch based - Analytic Hierarchy Process (PB-AHP) can monitor locust habitat. The overall accuracy reached 88%, which is 10% higher than the result based on the Analytic Hierarchy Process(AHP). These results show that the Patch based - Analytic Hierarchy Process (PB-AHP) model has strong robustness and generalization ability in identifying locust habitat and can provide scientific guidance for locust monitoring and control.

The majority of long noncoding RNAs (lncRNAs) contain transposable elements (TEs). PAHAL, a nuclear-retained lncRNA that is inserted by a Gypsy retrotransposon, has been shown to be a vital regulator of phenylalanine hydroxylase (PAH) gene expression that controls dopamine biosynthesis and behavioural aggregation in the migratory locust. However, the role of the Gypsy retrotransposon in the transcriptional regulation of PAHAL remains unknown. Here, we identified a Gypsy retrotransposon (named Gypsy element) as an inverted long terminal repeat located in the 3′ end of PAHAL, representing a feature shared by many other lncRNAs in the locust genome. The embedded Gypsy element contains a RNA nuclear localization signal motif, which promotes the stable accumulation of PAHAL in the nucleus. The Gypsy element also provides high-affinity SRSF2 binding sites for PAHAL that induce the recruitment of SRSF2, resulting in the PAHAL-mediated transcriptional activation of PAH. Thus, our data demonstrate that TEs provide discrete functional domains for lncRNA organization and highlight the contribution of TEs to the regulatory significance of lncRNAs.

The migratory locust Locusta migratoria (Orthoptera, Acrididae) is one of the most important pests due to its extensive and severe damage to crops in large parts of Africa and Asia. Biodegradable and ecologically natural products such as botanical insecticides are emerging candidates for replacement of usually applied chemical pesticides. The crude methanolic extract of Pergularia tomentosa (PME) was investigated for their toxicity and physiological aspects on L. migratoria nymphs. Results showed that treatment of newly emerged fourth and fifth instar nymphs resulted in significant mortality and significant repellent activity with an LC50 value of 0.18 and 0.38%, respectively, after seven days of treatment. The PME toxicity was also demonstrated by histopathological changes in the alimentary canal resulting in considerable disorganization and severe damage of the caeca and proventriculus structure. The extract induced cellular immune reactions which manifested by a significant decrease in the number of the differential haemocyte counts (prohemocytes and plasmatocytes) and important cell lysis. Data of biochemical analyses showed that the PME reduced the activity of acetylcholinesterase and induced the glutathione S-transferases. The neurotoxic effect was confirmed by the histological alterations in the brain structure, particularly in the neurosecretory cells showing typical signs of cell necrosis.

Maternal photoperiodic response is a key factor that affects offspring diapause in migratory locust, Locusta migratoria L. (Orthoptera: Acridoidea). Although many aspects of insect diapause have been studied, little is known about the molecular mechanisms of maternal photoperiodic response that influence diapause regulation. To gain insight into the possible mechanisms of maternal photoperiod influence on diapause regulation, proteomics data by label-free quantification analysis were generated from non-diapause and diapause eggs. A total of 175 proteins were differentially expressed between diapause and non-diapause eggs. Among them, 24 proteins were upregulated, and 151 proteins were downregulated. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichments were performed on all differentially expressed proteins (DEPs) and showed that peroxisome, insect hormone biosynthesis, and longevity regulating pathway may be related to diapause of migratory locust. Furthermore, we used qRT-PCR to verify some results of the proteomic analysis. Proteins such as hexamerin-like protein 4, juvenile hormone epoxide hydrolase 1 (JHEH1), cytochrome P450 and heat shock protein (HSP) 20.7 were predicted to be involved in diapause. This study provides an important reference for future research that will explore the mechanisms of diapause induced by maternal effects in migratory locust.

The genus Weissella is attracting an increasing amount of attention because of its multiple functions and probiotic potential. In particular, the species Weissella confusa is known to have great potential in industrial applications and exhibits numerous biological functions. However, the knowledge on this bacterium in insects is not investigated. Here, we isolated and identified W. confusa as the dominant lactic acid bacteria in the gut of the migratory locust. We named this strain W. confusa LM1, which is the first genome of an insect-derived W. confusa strain with one complete chromosome and one complete plasmid. Among all W. confusa strains, W. confusa LM1 had the largest genome. Its genome was the closest to that of W. confusa 1001271B_151109_G12, a strain from human feces. Our results provided accurate evolutionary relationships of known Weissella species and W. confusa strains. Based on genomic analysis, the pan-genome of W. confusa is in an open state. Most strains of W. confusa had the unique genes, indicating that these strains can adapt to different ecological niches and organisms. However, the variation of strain-specific genes did represent significant correlations with their hosts and ecological niches. These strains were predicted to have low potential to produce secondary metabolites. Furthermore, no antibiotic resistance genes were identified. At the same time, virulence factors associated with toxin production and secretion system were not found, indicating that W. confusa strains were not sufficient to perform virulence. Our study facilitated the discovery of the functions of W. confusa LM1 in locust biology and their potential application to locust management.

Vitellogenin receptor (VgR) plays a pivotal role in ovarian vitellogenin (Vg) uptake and vertical transmission of pathogenic microbes and Wolbachia symbionts. However, the regulatory mechanisms of VgR action as an endocytic receptor and translocation from oocyte cytoplasm to the membrane remain poorly understood. Here, by using the migratory locust Locusta migratoria as a model system, we report that juvenile hormone (JH) promotes VgR phosphorylation at Ser(1361) in the second EGF-precursor homology domain. A signaling cascade including GPCR, PLC, extracellular calcium, and PKC-ι is involved in JH-stimulated VgR phosphorylation. This posttranslational regulation is a prerequisite for VgR binding to Vg on the external surface of the oocyte membrane and subsequent VgR/Vg endocytosis. Acidification, a condition in endosomes, induces VgR dephosphorylation along with the dissociation of Vg from VgR. Phosphorylation modification is also required for VgR recycling from oocyte cytoplasm to the membrane. Additionally, VgR phosphorylation and its requirement for Vg uptake and VgR recycling are evolutionarily conserved in other representative insects including the cockroach Periplaneta americana and the cotton bollworm Helicoverpa armigera. This study fills an important knowledge gap of low-density lipoprotein receptors in posttranslational regulation, endocytosis, and intracellular recycling.

Simple Summary The demand for more sustainable protein sources is growing. Insects are believed to have potential as a sustainable protein source since they can be produced with lower environmental impact than current livestock. Furthermore, they have proven to be able to convert low-nutrient biomass into own nutrient-rich body mass, making them nutritionally interesting for applications such as food, feed and technical applications. The aim of this review was to provide more insight on the valorisation potential of agricultural side streams in feed for the rearing of yellow mealworm (Tenebrio molitor), house cricket (Acheta domesticus) and migratory locust (Locusta migratoria). Several key aspects need to be considered when aiming to valorise side streams in insect feed. The following aspects are discussed in detail: European legislation, the insects’ dietary requirements, the nutritional composition of insects and the availability of agricultural side streams in the EU. Additionally, research focusing on the production of insects on side streams is reviewed. Through this review, it is concluded that there is a huge potential to valorise residues in feed for the insects of interest. However, it is not a self-evident process. Abstract Due to increasing welfare and population, the demand for alternative protein sources, obtained with minimal use of natural resources, is rising in today’s society. Insects have the potential to be used as an alternative protein source since they are considered to be able to convert low-value biomass into high-value components, resulting in opportunities for valorisation of organic side streams. Moreover, insects are suggested to be a sustainable protein source, referring to the efficient “feed to body” mass conversion potential. The aim of this review was to explore the potential to rear the yellow mealworm (Tenebrio molitor), the house cricket (Acheta domesticus) and the migratory locust (Locusta migratoria) on low or not yet valorised organic side streams within the food supply chain. This was performed by collecting research information focusing on the rearing of the insects in scope on organic biomass. In addition, the nutritional composition of the produced insects as well as their dietary requirements will be reviewed. Finally, the availability of side streams in the EU will be discussed as well as their potential to be used as insects feed.

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Food and Food Allergens (NDA) was asked to deliver an opinion on the safety of frozen and dried formulations from migratory locust (Locusta migratoria) as a novel food pursuant to Regulation (EU) 2015/2283. The term migratory locust refers to the adult of the insect species Locusta migratoria. The NF is proposed in three formulations i) frozen without legs and wings; ii) dried without legs and wings; iii) ground with legs and wings. The main components of the NF are protein, fat and fibre (chitin) in the dried form of the NF, and water, protein, fat and fibre (chitin) in the frozen form of the NF. The Panel notes that the concentration of contaminants in the NF depends on the occurrence levels of these substances in the insect feed. The Panel notes that there are no safety concerns regarding the stability of the NF if the NF complies with the proposed specification limits during its entire shelf-life. The NF has a high protein content, although the true protein levels in the NF are overestimated when using the nitrogen to-protein conversion factor of 6.25, due to the presence of non-protein nitrogen from chitin. The applicant proposed to use the NF as frozen, dried and ground in the form of snack, and as a food ingredient in a number of food products. The target population proposed by the applicant is the general population. The Panel notes that considering the composition of the NF and the proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous. The submitted history of use and toxicity studies from literature did not raise safety concerns. The Panel considers that the consumption of the NF might trigger primary sensitisation to L. migratoria proteins and may cause allergic reactions in subjects with allergy to crustaceans, mites and molluscs. Additionally, allergens from the feed may end up in the NF. The Panel concludes that the NF is safe under the proposed uses and use levels. (C) 2021 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority.

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Food and Food Allergens (NDA) was asked to deliver an opinion on the safety of frozen and dried formulations from migratory locust (Locusta migratoria) as a novel food pursuant to Regulation (EU) 2015/2283. The term migratory locust refers to the adult of the insect species Locusta migratoria. The NF is proposed in three formulations i) frozen without legs and wings; ii) dried without legs and wings; iii) ground with legs and wings. The main components of the NF are protein, fat and fibre (chitin) in the dried form of the NF, and water, protein, fat and fibre (chitin) in the frozen form of the NF. The Panel notes that the concentration of contaminants in the NF depends on the occurrence levels of these substances in the insect feed. The Panel notes that there are no safety concerns regarding the stability of the NF if the NF complies with the proposed specification limits during its entire shelf‐life. The NF has a high protein content, although the true protein levels in the NF are overestimated when using the nitrogen‐to‐protein conversion factor of 6.25, due to the presence of non‐protein nitrogen from chitin. The applicant proposed to use the NF as frozen, dried and ground in the form of snack, and as a food ingredient in a number of food products. The target population proposed by the applicant is the general population. The Panel notes that considering the composition of the NF and the proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous. The submitted history of use and toxicity studies from literature did not raise safety concerns. The Panel considers that the consumption of the NF might trigger primary sensitisation to L. migratoria proteins and may cause allergic reactions in subjects with allergy to crustaceans, mites and molluscs. Additionally, allergens from the feed may end up in the NF. The Panel concludes that the NF is safe under the proposed uses and use levels.

The water extract of desert locust, Schistocerca gregaria, frass collected in the wild had an oviposition inhibitory (OI) effect when mixed with sand and presented to adults. Likewise, the leaves of six plant species, as well as frass produced by desert locusts fed with these plants, exerted OI effects when compared with the control sand wetted with water alone. In general, frass extracts had a greater OI effect than the extracts of leaves. The OI effect was also observed when adult desert locusts were exposed to extracts of frass produced by two other locusts, the Bombay locust, Nomadacris succincta, and the migratory locust, Locusta migratoria, fed with rescue grass, Bromus catharticus. Among the three species of locust, desert locust and migratory locust frass exerted a greater OI effect than Bombay locust frass. Frass samples extracted with hot and cool water produced similarly high OI effects, indicating that bacterial involvement during extraction is unlikely. Hatching rates of desert locusts were significantly reduced by extracts of all of the above mentioned frass when the extracts were mixed with sand and used to incubate the eggs. In contrast, the lethal effects of leaf extracts on the hatching rates varied depending on the plant species. The embryos became deformed within four days when three-day-old eggs were incubated in sand containing frass extracts from desert locusts fed with rescue grass, whereas no apparent morphological changes were observed when seven-day-old eggs were similarly tested, although their hatching rate was significantly reduced.

Eggs of the migratory locust, Locusta migratoria (Orthoptera: Acrididae), hatch synchronously when in a pod, but only sporadically when kept separately. Here, we aimed to detect the vibrational stimuli emitted by eggs that initiate synchronous hatching. First, we recorded the vibrations emitted by an egg pod and single eggs. One bout of vibrations consisted of 2 to 46 vibrations. The total number and amplitude of vibrations in single eggs increased as the time to hatch decreased. Eggs kept separately were continuously subjected during the last 2 days before hatching to recordings of vibrations from a single egg. Recordings made during the last 2.5 h before hatching caused these eggs to hatch significantly earlier than those not subjected to this treatment, the control. In contrast, eggs subjected to recordings made 8 to 10 h before hatching significantly delayed their hatching relative to the controls, which indicates that synchronous hatching of eggs is induced by age-dependent changes in vibrations from neighbouring eggs. Exposure to one large bout of vibrations (consisting of 40 vibrations in 101 s) was sufficient to induce synchronous hatching in eggs kept separately when applied 5 h before hatching, but not 36 or 11.5 h before hatching. Visual inspection of the spectra indicated that the vibrations had two peaks at about 100 Hz and 1.5 kHz. Only exposure to the latter altered the hatching time of eggs. The embryo moved the posterior part of its abdomen when emitting the vibrations. These results indicate that the synchronous hatching of locust eggs is induced when the embryos emit particular vibrations.

Phenotypic plasticity plays fundamental roles in successful adaptation of animals in response to environmental variations. Here, to reveal the transcriptome reprogramming in locust phase change, a typical phenotypic plasticity, we conducted a comprehensive analysis of multiple phase-related transcriptomic datasets of the migratory locust. We defined PhaseCore genes according to their contribution to phase differentiation by the adjustment for confounding principal components analysis algorithm (AC-PCA). Compared with other genes, PhaseCore genes predicted phase status with over 87.5% accuracy and displayed more unique gene attributes including the faster evolution rate, higher CpG content and higher specific expression level. Then, we identified 20 transcription factors (TFs) named PhaseCoreTF genes that are associated with the regulation of PhaseCore genes. Finally, we experimentally verified the regulatory roles of three representative TFs (Hr4, Hr46, and grh) in phase change by RNAi. Our findings revealed that core transcriptional signatures are involved in the global regulation of locust phase changes, suggesting a potential common mechanism underlying phenotypic plasticity in insects. The expression and network data are accessible in an online resource called LocustMine (http://www.locustmine.org:8080/locustmine). Electronic supplementary material The online version of this article (10.1007/s13238-019-0648-6) contains supplementary material, which is available to authorized users.

The efficacy of Agroecosystem Compounds, fipronil, spinosad and chlorantraniliprole as alternatives to the conventional insecticides on the mortality rates and haemolymph protein, lipid and carbohydrate contents were assessed in Schistocerca gregaria and Locusta migratoria migratorioides. One day old 5th nymphal instars of each of the mentioned species were treated by various concentrations of tested compounds by the feeding technique. The results indicated that Locusta migratoria migratorioides is more sensitive to all tested insecticides than Schistocerca gregaria. In both species, fipronil was the most effective followed by spinosad, and chlorantraniliprole was the least effective. LC30 concentrations of the three compounds were used to treat the 5th instar hoppers of both species to examine the effects of sub lethal concentrations of the mentioned insecticides on the haemolymph total proteins, total lipids and total carbohydrates. It was found that haemolymph proteins, lipids and carbohydrates contents were dramatically reduced for the treated hoppers when compared to the untreated ones of both species. Malformed hopper and wrinkled winged adult were seen in insects treated by sub lethal concentrations of fipronil and spinosad.

This feasibility study reports the use of direct analysis in real-time high-resolution mass spectrometry (DART-HRMS) in profiling the powders from edible insects, as well as the potential for the identification of different insect species by classification modeling. The basis of this study is the revolution that has occurred in the field of analytical chemistry, with the improved capability of ambient mass spectrometry to authenticate food matrices. In this study, we applied DART-HRMS, coupled with mid-level data fusion and a learning method, to discriminate between Acheta domesticus (house cricket), Tenebrio molitor (yellow mealworm), Locusta migratoria (migratory locust), and Bombyx mori (silk moth). A distinct metabolic fingerprint was observed for each edible insect species, while the Bombyx mori fingerprint was characterized by highly abundant linolenic acid and quinic acid; palmitic and oleic acids are the statistically predominant fatty acids in black soldier fly (Hermetia illucens). Our chemometrics also revealed that the amino acid proline is a discriminant molecule in Tenebrio molitor, whereas palmitic and linoleic acids are the most informative molecular features of the house cricket (Acheta domesticus). Good separation between the four different insect species was achieved, and cross-validation gave 100% correct identification for all training samples. The performance of the random forest classifier was examined on a test set and produced excellent results, in terms of overall accuracy, sensitivity, and specificity. These results demonstrate the reliability of the DART-HRMS as a screening method in a future quality control scenario to detect complete substitution of insect powders.

Background The migratory locust Locusta migratoria is one of the most important agricultural pests worldwide. The nuclear export factor 1 (NXF1) protein plays a crucial role in mediating mRNA transport from the nucleus to the cytoplasm. This study evaluates whether NXF1 could be a potential target for RNAi-mediated pest control of L. migratoria. Results We cloned and characterized the nuclear export factor lm-nxf1 of L. migratoria. Lm-nxf1 was expressed in all tissues examined, including head, fat body, hemolymph, trunk, leg and midgut, with high expression observed in the hemolymph and fat body. Injection of lm-nxf1 dsRNA into hemolymph resulted in inhibition of mRNA export in hemocytes, which were used as a target for observing mRNA export. Total hemocyte levels were reduced by ca. 97% in lm-nxf1-dsRNA-treated locusts, and high insect mortality occurred with LT50 = 7.75 day as compared with 18.15 day for gfp-dsRNA-treated controls. Further, the locust intestine became atrophy, and the opportunistic pathogens Enterobacter aerogenes, Klebsiella pneumoniae and Enterobacter asburiae were specifically detected in midgut after lm-nxf1 dsRNA treatment. Conclusions The results reveal that knockdown of the lm-nxf1 gene affects the survival of L. migratoria, indicating that lm-nxf1 is a potential target for RNAi-mediated pest control. © 2018 Society of Chemical Industry.

Juvenile hormone (JH) is known to promote cell polyploidization for insect vitellogenesis and egg production, but the underlying mechanisms remain poorly understood. Using the migratory locust Locusta migratoria as a model system, we report here that the expression of cyclin-dependent kinase 6 (Cdk6) and adenovirus E2 factor-1 (E2f1), the core mediators in cell cycle progression is regulated by JH and its receptor Methoprene-tolerant (Met). JH acts through its receptor complex comprised of Met and Taiman to directly activate the transcription of Cdk6 and E2f1. Depletion of Cdk6 or E2f1 results in significantly decreased ploidy, precocious mitotic entry and increased cell numbers in the fat body, accompanied by substantial reduction of Vitellogenin gene expression, blocked ovarian growth and arrested oocyte maturation. These findings indicate a crucial role of Cdk6 and E2f1 in JH-regulated polyploidization and vitellogenesis as well as a novel regulatory machinery for endocycling in insects.

Background Non-Drosophila insects provide diverse aging types and important complementary systems for studies of aging biology. However, little attention has been paid to the special roles of non-Drosophila insects in aging research. Here, the aging-related features of the migratory locust, Locusta migratoria, were determined at the physiological, cellular, and transcriptional levels. Results In physiological assessments, the flight performance and sperm state of locusts displayed clear aging-related decline in male adults. Transcriptional analyses demonstrated locusts have similar aging-related genes with model species. However, different from those of Drosophila and mammals, the organ-specific aging transcriptional features of locusts were characterized by intensive expression changes in flight muscle and fat body and little transcriptional changes in brain. The predominant transcriptional characteristics of flight muscle and fat body aging were changes in expression of mitochondrion-related genes and detoxification and phagocytosis genes, respectively. Cellular assessments revealed the incidence of mitochondrial abnormalities significantly increased in aged flight muscle, and apoptotic signals and nuclear abnormalities were enhanced in aged fat body but not in brain. In addition, some well-known aging genes and locust aging-related genes (i.e., IAP1, PGRP-SA, and LIPT1), whose roles in aging regulation were rarely reported, were demonstrated to affect lifespan, metabolism, and flight ability of locusts after RNAi. Conclusion This study revealed multi-level aging signatures of locust, thus laying a foundation for further investigation of aging mechanisms in this famous insect in the future.

Simple Summary It was reported that insulin-like and fork head transcription factor (FOXO) are involved in the regulation of diapause in insects. However, the upstream modulators of the insulin-like signaling pathway (ISP) involved in diapause regulation are still unknown. We used RNAi and an inhibitor to treat PTK and PTP1B in adult tissues and injected Prx V protein or RNAi Prx V under both short and long photoperiod conditions to identify both proteins and broader cellular metabolism influences on diapause regulation. We found that under short photoperiod conditions PTP1B in female adults induces egg diapause, whereas PTK in female adults inhibits egg diapause. Intriguingly, we also found that the antioxidant enzyme Prx V is a negative regulator of NADPH oxidizing reaction, and apparently decreases reactive oxygen species (ROS) production and NADPH-OX activity. Thus, these results indicate that PTP1B, PTK and Prx V are upstream modulators that regulate diapause in eggs via the insulin signaling pathway. Furthermore, these findings have revealed a possible bridge connecting diapause hormone signaling to the insulin-like signaling pathway. Abstract Diapause is a physiological development arrest state that helps insects to adapt to seasonality and overcome adverse environmental conditions. Numerous reports have indicated that insulinlike and fork head transcription factor (FOXO) are involved in the regulation of diapause in insects. However, the upstream modulators of the insulin-like signaling pathway (ISP) involved in diapause regulation are still unknown. Here, we used RNAi and an inhibitor to treat PTK and PTP1B in adult tissues and injected Prx V or RNAi Prx V under both short and long photoperiod conditions and monitored effects on the expression of ISP genes, the phosphorylation levels for IR and IRS, the activity of NADPH oxidase, the accumulation of reactive oxygen species (ROS) and energy metabolism, seeking to identify both proteins and broader cellular metabolism influences on diapause regulation. We found that under short photoperiod conditions PTP1B in female adults induces egg diapause, whereas PTK in female adults inhibits egg diapause. Intriguingly, we also found that the antioxidant enzyme Prx V is a negative regulator of NADPH oxidizing reaction and apparently decreases ROS production and NADPH-OX activity. In contrast, all the eggs laid by adults that were treated with a series of knockdown or purified-protein injection experiments or inhibitor studies and that were reared under long photoperiod conditions hatched successfully. Thus, our results suggest a mechanism wherein diapause-related proteins (PTP1B, PTK, and Prx V) of female adults are the upstream modulators that regulate offspring eggs’ diapause process through the insulin-like signaling pathway under short photoperiod conditions.

Locusts are advantageous organisms to elucidate mechanisms of olfactory coding at the systems level. Sensory input is provided by the olfactory receptor neurons of the antenna, which send their axons into the antennal lobe. So far, cellular properties of neurons isolated from the circuitry of the olfactory system, such as transmitter-induced calcium responses, have not been studied. Biochemical and immunocytochemical investigations have provided evidence for acetylcholine as classical transmitter of olfactory receptor neurons. Here, we characterize cell cultured projection and local interneurons of the antennal lobe by cytosolic calcium imaging to cholinergic stimulation. We bulk loaded the indicator dye Cal-520 AM in dissociated culture and recorded calcium transients after applying cholinergic agonists and antagonists. The majority of projection and local neurons respond with increases in calcium levels to activation of both nicotinic and muscarinic receptors. In local interneurons, we reveal interactions lasting over minutes between intracellular signaling pathways, mediated by muscarinic and nicotinic receptor stimulation. The present investigation is pioneer in showing that Cal-520 AM readily loadsLocusta migratorianeurons, making it a valuable tool for future research in locust neurophysiology, neuropharmacology, and neurodevelopment.

Background Diapause plays a vital role in the survival and population reproduction of the migratory locust in temperate regions. Although some studies have suggested that photoperiod and temperature are two important environmental factors involved in the induction and termination of embryonic diapause in locusts, the independent or synergetic roles of these two factors are still unclear. Results We designed crossover experiments to investigate the effects of photoperiod, temperature and treatment time on the induction and termination of embryonic diapause in locusts by optical coherence tomography (OCT), which can track the entire embryonic developmental process via noninvasive 3D real-time imaging. Diapause induction experiments showed that a short photoperiod (in adults) and a low egg treatment temperature are the most important prerequisites for inducing egg diapause. The diapause rate reached 90.7% when the adults were reared under a short photoperiod (8 L:16 D) and the eggs were treated at 22 °C. However, the adult rearing temperature had no significant effect on diapause induction. Analysis of variance also confirmed that there was a significant interactive effect between parental photoperiod and egg treatment temperature. Moreover, diapause termination experiments showed that 13 °C (near the developmental threshold temperature of 15 °C) for 20 days is the most effective condition for terminating diapause. Conclusion Our study provides accurate photoperiod and temperature thresholds for inducing and terminating diapause in the migratory locust, suggesting potential applications in the prediction and control of locust plagues. © 2021 Society of Chemical Industry.

The increasing development of edible insect flours as alternative sources of proteins added to food and feed products for improving their nutritional value, necessitates an accurate evaluation of their possible adverse side-effects, especially for individuals suffering from food allergies. Using a proteomic- and bioinformatic-based approach, the diversity of proteins occurring in currently consumed edible insects such as silkworm (Bombyx mori), cricket (Acheta domesticus), African migratory locust (Locusta migratoria), yellow mealworm (Tenebrio molitor), red palm weevil (Rhynchophorus ferrugineus), and giant milworm beetle (Zophobas atratus), was investigated. Most of them consist of phylogenetically-related protein allergens widely distributed in the different groups of arthropods (mites, insects, crustaceans) and mollusks. However, a few proteins belonging to discrete protein families including the chemosensory protein, hexamerin, and the odorant-binding protein, emerged as proteins highly specific for edible insects. To a lesser extent, other proteins such as apolipophorin III, the larval cuticle protein, and the receptor for activated protein kinase, also exhibited a rather good specificity for edible insects. These proteins, that are apparently missing or much less represented in other groups of arthropods, mollusks and nematods, share well conserved amino acid sequences and very similar three-dimensional structures. Owing to their ability to trigger allergic responses in sensitized people, they should be used as probes for the specific detection of insect proteins as food ingredients in various food products and thus, to assess their food safety, especially for people allergic to edible insects.

Outbreaks of locust plagues result from the long-term accumulation of high-density egg production. The migratory locust, Locusta migratoria, displays dramatic differences in the egg-laid number with dependence on population density, while solitarious locusts lay more eggs compared to gregarious ones. However, the regulatory mechanism for the egg-laid number difference is unclear. Herein, we confirm that oosorption plays a crucial role in the regulation of egg number through the comparison of physiological and molecular biological profiles in gregarious and solitarious locusts. We find that gregarious oocytes display a 15% higher oosorption ratio than solitarious ones. Activinβ (Actβ) is the most highly upregulated gene in the gregarious terminal oocyte (GTO) compared to solitarious terminal oocyte (STO). Meanwhile, Actβ increases sharply from the normal oocyte (N) to resorption body 1 (RB1) stage during oosorption. The knockdown of Actβ significantly reduces the oosorption ratio by 13% in gregarious locusts, resulting in an increase in the egg-laid number. Based on bioinformatic prediction and experimental verification, microRNA-34 with three isoforms can target Actβ. The microRNAs display higher expression levels in STO than those in GTO and contrasting expression patterns of Actβ from the N to RB1 transition. Overexpression of each miR-34 isoform leads to decreased Actβ levels and significantly reduces the oosorption ratio in gregarious locusts. In contrast, inhibition of the miR-34 isoforms results in increased Actβ levels and eventually elevates the oosorption ratio of solitarious locusts. Our study reports an undescribed mechanism of oosorption through miRNA targeting of a TGFβ ligand and provides new insights into the mechanism of density-dependent reproductive adaption in insects., Author summary The continuous accumulation of high-density eggs laid by flying swarms of adults results in huge populations of flightless juveniles, which contributes to the outbreaks of locust plagues. An interesting phenomenon is that locusts have the phenotypic plasticity of reproduction. The gregarious locusts lay fewer big eggs than do solitarious phase locusts. In contrast, the solitarious phase locusts lay more small eggs compared to the gregarious locusts. We find the egg-laid number is not only regulated by the phase status of parents but also controlled by oosorption, a type of oocyte death. Further studies confirmed the phase-related ratio of oocyte death in the mother is regulated by a microRNA, which posttranscriptionally influences the expression level of a TGFβ ligand. This maternal effect on progeny size is especially critical for gregarious locusts to control the population size and maintain population fitness, and for solitarious locusts to enhance chance for gregarization and further enlargement of population size. This is the first study to reveal the molecular mechanism underlying the regulation of a microRNA-gene circuit for locust oocyte death to determine the offspring number. These findings can provide some important clues to develop potential drugs to prevent vast locust reproduction from a plague upsurge.

Background Core promoters have a substantial influence on various steps of transcription, including initiation, elongation, termination, polyadenylation, and finally, translation. The characterization of core promoters is crucial for exploring the regulatory code of transcription initiation. However, the current understanding of insect core promoters is focused on those of Diptera (especially Drosophila) species with small genome sizes. Results Here, we present an analysis of the transcription start sites (TSSs) in the migratory locust, Locusta migratoria, which has a genome size of 6.5 Gb. The genomic differences, including lower precision of transcription initiation and fewer constraints on the distance from transcription factor binding sites or regulatory elements to TSSs, were revealed in locusts compared with Drosophila insects. Furthermore, we found a distinct bimodal log distribution of the distances from the start codons to the core promoters of locust genes. We found stricter constraints on the exon length of mRNA leaders and widespread expression activity of the distant core promoters in locusts compared with fruit flies. We further compared core promoters in seven arthropod species across a broad range of genome sizes to reinforce our results on the emergence of distant core promoters in large-sized genomes. Conclusions In summary, our results provide novel insights into the effects of genome size expansion on distant transcription initiation.

Time series of abundance indices for Desert Locusts Schistocerca gregaria (Forsk&aring, l 1775) and Oriental Migratory Locusts Locusta migratoriamanilensis (Meyen 1835) were analysed independently and in relation to measures of solar activity and ocean oscillation systems. Data were compiled on the numbers of territories infested with swarms of the Desert Locust from 1860&ndash, 2015 and an inferred series that compensated for poor reporting in the 1860 to 1925 period. In addition, data for 1930 to 2014, when reports are considered to have been consistently reliable were converted to numbers of 1&deg, grid squares infested with swarms and separated according to four different geographical regions. Spectral analysis to test the hypothesis that there are cycles in the locust dynamics revealed periodicities of 7.5 and 13.5 years for the inferred series that were significant according to the Ornstein-Uhlenbeck state-space (OUSS) test. Similar periodicities were evident in the 1&deg, grid square data and in each of the regions but even though these were significantly different from white noise, they were not significant according to the OUSS criterion. There were no significant peaks in the Oriental Migratory Locust results with the OUSS test, but the data were significantly different from white noise. To test hypotheses that long term trends in the locust dynamics are driven by solar activity and/or oceanic oscillation systems (the Southern Oscillation Index (SOI), the North Atlantic Oscillation Index (NAO) and the Indian Ocean Dipole (IOD)), the original locust data series and their Kalman-filtered low frequency (LF) components were tested for causality using both spectral coherence tests and convergent cross mapping. Statistically significant evidence was found that solar activity measured by numbers of sunspot groups drive the dynamics, especially the LF components, of both species. In addition, causal links were inferred between both the SOI and NAO data and Desert Locust dynamics. Spectral coherence was also found between sunspot groups and the NAO, the IOD and LF SOI data. The data were also analysed showing that the LF SOI had causal links with the LF inferred Desert Locust series. In addition, the LF NAO was causally linked to the LF 1&deg, grid square data, with the NAO for December-March being most influential. The results suggest that solar activity plays a role in driving locust abundance, but that the mechanisms by which this happens, and whether they are mediated by fluctuations in oceanic systems, is unclear. Furthermore, they offer hope that information on these phenomena might enable a better early warning forecasting of Desert Locust upsurges.

Chitin deacetylases including CDA1 and CDA2, containing a chitin deacetylase domain and an LDL domain, have been reported to be essential for cuticle structure differentiation in different insect species. However, it is yet unexplored whether CDA1 and CDA2 activity is needed for the function of the cuticle as a barrier against pathogen and xenobiotics penetration. In this study, we studied the efficiency of fungal infection in the migratory locust Locusta migratoria in dependence of LmCDA1 and LmCDA2 function. Second instar nymphs injected with dsRNA against LmCDA1 and LmCDA2 transcripts were less resistant against the infection by the fungus Metarhizium anisopliae than control nymphs. At the same time, permeability to organophosphorus pesticides was increased in these nymphs. Interestingly, the CHC amounts at the cuticle surface were unaffected upon LmCDA1 and LmCDA2 reduction. These results suggest that the barrier function of the locust cuticle not only depends on surface CHCs, but also on an intact procuticle.

There is a growing appreciation that insect distribution and abundance are associated with the limits of thermal tolerance, but the physiology underlying thermal tolerance remains poorly understood. Many insects, like the migratory locust (Locusta migratoria), suffer a loss of ion and water balance leading to hyperkalaemia (high extracellular [K+]) in the cold that indirectly causes cell death. Cells can die in several ways under stress, and how they die is of critical importance to identifying and understanding the nature of thermal adaptation. Whether apoptotic or necrotic cell death pathways are responsible for low-temperature injury is unclear. Here, we use a caspase-3 specific assay to indirectly quantify apoptotic cell death in three locust tissues (muscle, nerves and midgut) following prolonged chilling and recovery from an injury-inducing cold exposure. Furthermore, we obtain matching measurements of injury, extracellular [K+] and muscle caspase-3 activity in individual locusts to gain further insight into the mechanistic nature of chilling injury. We found a significant increase in muscle caspase-3 activity, but no such increase was observed in either nervous or gut tissue from the same animals, suggesting that chill injury primarily relates to muscle cell death. Levels of chilling injury measured at the whole animal level, however, were strongly correlated with the degree of haemolymph hyperkalaemia, and not apoptosis. These results support the notion that cold-induced ion balance disruption triggers cell death but also that apoptosis is not the main form of cell damage driving low-temperature injury.

The first list of the rare grasshoppers of the Baraba and Kulunda steppes is presented. Two sets of distribution data are compared: (1) for the first half of 20th century and (2) for 1972–2019. A series of digital maps was generated by MapInfo 12.03. The distribution patterns of several species, namely Asiotmethis muricatus (Pallas), Notostaurus albicornis (Eversmann), Eremippus simplex (Eversmann), Myrmeleotettix antennatus (Fieber), Gomphocerippus rufus (Linnaeus), Mesasippus arenosus (Bey-Bienko), Mecostethus parapleurus (Hagenbach), Locusta migratoria Linnaeus, did not change significantly. Four taxa (Asiotmethis jubatus (Uvarov), Arcyptera fusca (Pallas), Stenobothrus carbonarius (Eversmann), Sphingonotus coerulipes Uvarov) were relatively often in the first half of 20th century and nowadays they are extremely rare. Two species, namely Megaulacobothrus aethalinus (Zubovsky) and Aeropedellus variegatus (Fischer de Waldheim), were recently found near the south-eastern and north-eastern boundaries of the region respectively. There are also the type localities of Asiotmethis jubatus and Mesasippus arenosus in the Kulunda steppe.

Simple Summary Locust plagues have been devastating vegetation and agriculture since ancient times. The tendency of locusts to form huge swarms and migrate long distances is a hallmark of the locust phenomenon. The ecological and economic importance of locust plagues has attracted much research into different aspects of the natural history of these insects. One of the many investigated aspects of the locust phenomenon is that of their interaction with the bacteria they harbor in and on their body. These locust-associated bacteria have been shown to affect certain physiological traits (such as immunity and swarm cohesion), as well as possibly affecting and being affected by different factors in the locust biology. Here, we review the current understanding of the locust–bacteria interaction. We focus on identifying the bacterial strains and their locations within the insects; the role of the bacteria and their importance to their host’s life; the mechanism of transmitting important bacteria across locust generations; and more. Finally, we offer some new perspectives and research directions that could broaden our understanding of the locust-associated bacteria and their role in locust outbreaks. Abstract As one of the world’s most infamous agricultural pests, locusts have been subjected to many in-depth studies. Their ability at one end of their behavioral spectrum to live as solitary individuals under specific conditions, and at the other end of the spectrum to form swarms of biblical scale, has placed them at the focus of vast research efforts. One important aspect of locust ecology is that of their interactions with the bacteria that reside in and on them. Although this aspect of locust ecology has been little studied relative to the mainstream locust research, these bacteria have been shown both to affect locust immunity and to participate in maintaining swarm integrity through the secretion of attractant volatiles. The interaction between locusts and their bacteria seems, however, to be bi-directional, with the bacteria themselves, as recently shown, being influenced by their host’s swarming tendencies. This seems to be a consequence of the bacterial composition in the locust’s gut, reproductive organs, and integument undergoing change with the change in their host’s behavior. In this review we describe the current state of knowledge of the locust–bacteria interactions (data exists mainly for the desert and the migratory locusts), as well as highlighting some newly-gained understanding; and offer perspectives for future research.

Sugarbabe is a C2 H2 zinc-finger transcription factor that is sensitive to sugar and essential for lipid biosynthesis in larvae of Drosophila melanogaster. However, the role of Sugarbabe in adult insect development remains unexplored. Vitellogenesis is a nutrient-dependent process that is promoted by juvenile hormone (JH) in many insect species. Here, we cloned an ortholog gene of D. melanogaster Sugarbabe (DmSug) in the migratory locust Locusta migratoria. The locust Sugarbabe (LmSug) has five C2 H2 zinc-finger motifs similar to DmSug. LmSug was expressed at a low level in adult female locusts raised under poor nutrient conditions. JH treatment increased the expression level of LmSug. Knockdown of the JH receptor gene Met caused a reduction of LmSug expression. Depletion of the LmSug transcript level caused a significant reduction in vitellogenin expression in the fat body, resulting in impaired oocyte development and ovary growth. The results suggest that LmSug is expressed in response to JH, and plays an essential role in female insect reproduction.

Apoptosis plays a major role in development, tissue renewal and the progression of degenerative diseases. Studies on various types of mammalian cells reported a pro-apoptotic function of acetylcholinesterase (AChE), particularly in the formation of the apoptosome and the degradation of nuclear DNA. While three AChE splice variants are present in mammals, invertebrates typically express two ache genes that code for a synaptically located protein and a protein with non-synaptic functions respectively. In order to investigate a potential contribution of AChE to apoptosis in insects, we selected the migratory locust Locusta migratoria. We established primary neuronal cultures of locust brains and characterized apoptosis progression in vitro. Dying neurons displayed typical characteristics of apoptosis, including caspase-activation, nuclear condensation and DNA fragmentation visualized by TUNEL staining. Addition of the AChE inhibitors neostigmine and territrem B reduced apoptotic cell death under normal culture conditions. Moreover, both inhibitors completely suppressed hypoxia-induced neuronal cell death. Exposure of live animals to severe hypoxia moderately increased the expression of ace-1 in locust brains in vivo. Our results indicate a previously unreported role of AChE in insect apoptosis that parallels the pro-apoptotic role in mammalian cells. This similarity adds to the list of apoptotic mechanisms shared by mammals and insects, supporting the hypothesized existence of an ancient, complex apoptosis regulatory network present in common ancestors of vertebrates and insects. Electronic supplementary material The online version of this article (10.1007/s10495-020-01630-4) contains supplementary material, which is available to authorized users.

Kr&uuml, ppel-homolog 1 (Kr-h1) is a zinc finger transcription factor maintaining the status quo in immature insect stages and promoting reproduction in adult insects through the transduction of the Juvenile Hormone (JH) signal. Knockdown studies have shown that precocious silencing of Kr-h1 in the immature stages results in the premature development of adult features. However, the molecular characteristics and reproductive potential of these premature adult insect stages are still poorly understood. Here we report on an adult-like or &lsquo, adultoid&rsquo, phenotype of the migratory locust, Locusta migratoria, obtained after a premature metamorphosis induced by the silencing of LmKr-h1 in the penultimate instar. The freshly molted adultoid shows precocious development of adult features, corresponding with increased transcript levels of the adult specifier gene LmE93. Furthermore, accelerated ovarian maturation and vitellogenesis were observed in female adultoids, coinciding with elevated expression of LmCYP15A1 in corpora allata (CA) and LmKr-h1 and vitellogenin genes (LmVg) in fat body, whereas LmE93 and Methoprene-tolerant (LmMet) transcript levels decreased in fat body. In adultoid ovaries, expression of the Halloween genes, Spook (LmSpo) and Phantom (LmPhm), was elevated as well. In addition, the processes of mating and oviposition were severely disturbed in these females. L. migratoria is a well-known, swarm-forming pest insect that can destroy crops and harvests in some of the world&rsquo, s poorest countries. As such, a better understanding of factors that are capable of significantly reducing the reproductive potential of this pest may be of crucial importance for the development of novel locust control strategies.

During prolonged or severe chilling, the majority of insects accrue chilling injuries that are typically quantified by scoring neuromuscular function after rewarming. In the cold, these chill susceptible insects, like the migratory locust (Locusta migratoria) suffer a loss of ion and water balance that is hypothesized to initiate cell death. Whether apoptotic or necrotic cell death pathways are responsible for this chilling injury is unclear. Here, we use a caspase-3 specific assay to indirectly quantify apoptosis in three locust tissues (muscle, nerves, and midgut) following prolonged chilling and recovery from an injury-inducing cold exposure. Furthermore, we obtain matching measurements of injury, hemolymph [K+], and muscle caspase-3 activity in individual locusts to gain further insight into mechanistic nature of chilling injury. We hypothesized that apoptotic cell death in both muscle and nerve tissue drives motor defects following cold exposure in insects, and that there would be a strong association between cold- induced injury, hyperkalemia, and muscle caspase-3 activity. We found a significant increase in muscle caspase-3 activity, but no such increase was observed in either nervous or gut tissue from the same animals, suggesting that chill injury primarily relates to apoptotic muscle cell death. However, the levels of chilling injury measured at the whole animal level prior to tissue sampling were strongly correlated with the degree of hemolymph hyperkalemia, but not apoptosis. These results support the notion that cold-induced ion balance disruption triggers cell death but also that apoptosis is not the main cell death pathway driving injury in the cold.Significance StatementTemperature has profound effects on animal fitness and sets limits to animal distribution. To understand and model insect responses to climate, we need to know how temperature sets limits to their survival. There is strong evidence that a collapse of ion and water balance occurs in insects in the cold, and it is generally held that the resulting cold injury is caused by activation of programmed cell death (apoptosis). Here, we directly test this idea and show for the first time that although the loss of ion balance is a strong predictor of individual survival outcomes, apoptosis is not the primary cause of cold-induced injury.

Apolipophorins are carrier proteins that bind lipids and mediate their transport from tissue to tissue in animals. Apolipophorin I and II (apoLp-II/I) are the major apolipophorins in insects. The implication of apoLp-II/I in cuticle lipid-barrier formation in insects has not been addressed to date. In the present study, we investigated the function of apoLp-II/I in the migratory locust Locusta migratoria (LmapoLp-II/I). During the development of fifth instar nymphs, LmapoLp-II/I transcript levels increased until mid-instar, and then decreased gradually until molting to the adult stage. We found that LmapoLp-II/I was predominately expressed in the fat body and the integument including oenocytes and epidermal cells. Immunodetection experiments revealed that LmapoLp-I mainly localized in the cytoplasm of oenocytes and epidermal cells. Silencing of LmapoLp-II/I caused molting defects in nymphs. Importantly, RNA interference against LmapoLp-II/I resulted in a significant decrease in the content of cuticle surface lipids including alkanes and methyl alkanes. Cuticular permeability was significantly enhanced in these nymphs in Eosin Y penetration assays. By consequence, desiccation resistance and insecticide tolerance of dsLmapoLp-II/I-treated locusts were reduced. Taken together, our results indicate that LmapoLp-II/I is involved in the transport and deposition of surface-cuticular lipids that are crucial for maintaining normal cuticle barrier function in L. migratoria.

The narrow host range entomopathogenic fungus, Metarhizium acridum, is an environmentally friendly acridid specific pathogen used for locust control. The locust is capable of responding within hours of infection, however, little is known concerning how the locust detects the pathogen. Here, we have identified 3213 proteins in the infected antennal proteome of the migratory locust, Locusta migratoria. iTRAQ comparative analyses of antennal proteomes identified 194 differentially abundant proteins (DAPs) between uninfected and infected males, 218 DAPs between uninfected and infected females, and 240 DAPs between infected males and infected females. In relation to olfaction, a total of 29 chemosensory proteins (CSPs), 9 odorant binding proteins (OBPs), 31 odorant receptors (ORs), and 8 ionotropic receptors (IRs) were differentially abundant after M. acridum infection, with a subset of 12 proteins found in both infected male and female antennae not present in uninfected individuals. The time course of the gene expression profiles of olfaction related DAPs were investigated by quantitative real-time PCR (qRT-PCR). Our data indicate significant changes in the antennal proteomes of male and female locusts in response to a microbial pathogen, highlighting the potential participation of olfactory processes in pathogen detection and response. Biological significance The ability of an organism to detect microbial pathogens is essential for mounting a response to mitigate the spread of the infection. Using iTRAQ-based proteomic analyses changes in the protein repertoire of the antennae of male and female locusts in response to infection by a host-specific pathogen were determined. These data show proteomic alterations that are also sex-specific, identifying members of olfactory pathways that are modified in response to infection. Our data identify antennal and related olfactory proteins that are candidates for mediating host detection of pathogens, and that may contribute to subsequent behavioral and/or immune responses of the host to the infection challenge.

Long noncoding RNAs (lncRNAs) regulate various biological processes from gene expression to animal behavior. Although protein-coding genes, microRNAs, and neuropeptides play important roles in the regulation of phenotypic plasticity in migratory locust, empirical studies on the function of lncRNAs in the process remain limited. Here, we applied high-throughput RNA-seq to characterize the expression patterns of lncRNAs and mRNAs in the time course of the locust phase change. LncRNAs displayed more rapid response at the early stages of the time-course treatments than mRNA expression. Functional annotations demonstrated that early changed lncRNAs employed different pathways in isolation and crowding processes to cope up with the changes in population density. Finally, two overlapping hub lncRNA loci in the crowding and isolation networks were screened to be functionally verified. Experimental validation indicated that LNC1010057 could act as a potential regulator to modulate the locust phase change. This work offers new insights into the mechanism underlying the locust phase change and expands the scope of lncRNA functions in animal behavior.

Mucins are highly glycosylated proteins that are characterized by a higher proportion of threonine, serine, and proline residues in their sequences. Although mucins in humans and vertebrates have been implicated in many biological processes, their roles in growth and development in invertebrates such as in insects remain largely unknown. Based on bioinformatic analyses, we identified eight mucin or mucin-like genes in the migratory locust, Locusta migratoria. RNA interference against these genes demonstrated that three Lmmucin genes were essential for the survival of L. migratoria nymphs, and one Lmmucin was required for adult wing development. Indeed, knockdown of Lmhemomucin and Lmmucin-12 caused lethal phenotypes, with an observed defect of the gastric caeca in which cells were detached from cell junctions. Deficiency of LmIIM3 resulted in lethality of nymphs, with defects of the peritrophic membrane in midgut. Suppression of Lmmucin-17 greatly impaired the structural integrity of the wing cuticle during nymph-adult molting. The present study revealed the significance of mucin and mucin-like genes in insect growth and development, using the orthopteran insect locust as a model.

Lipids of the insect cuticle have important roles in resistance against the arid environment and invasion of foreign substances. Fatty acid synthase (FAS) is an important enzyme of the insect lipid synthesis pathway. In the present study, we identified three FAS genes from transcriptome data of the migratory locust, Locusta migratoria, based on bioinformatics analyses. Among them, two FAS genes (LmFAS1 and LmFAS3) are highly expressed in the integument of fifth instar nymphs. Suppression of LmFAS1 and LmFAS3 by RNA interference caused lethality during ecdysis or shortly after moulting. The weight of the locusts and the content of lipid droplets were reduced compared with those of the control. The results of gas chromatography-mass spectrometry analysis showed that knockdown of LmFAS3 led to a decrease of both cuticular hydrocarbons and inner hydrocarbons (CHCs and IHCs) contents, especially the content of methyl branched hydrocarbons. By contrast, knockdown of LmFAS1 only resulted in a decrease in the IHC content, but not that of CHCs. By consequence, in LmFAS1- and LmFAS3-suppressed locusts, hydrocarbon deficiency reduced desiccation resistance and enhanced cuticle permeability and sensitivity to insecticides. These results indicate that LmFAS1 and LmFAS3 are essential for hydrocarbon production and cuticle permeability, which play influential roles in waterproofing the insect cuticle.