Insects, Volume 15, Issue 1 (January 2024) – 79 articles

The populations of clearwing moth borers in Korea have recently caused extensive and severe damage to pin oaks (Quercus palustris Munchh.). We conducted field monitoring and molecular analyses to identify them in an integrated manner. Morphological examination and molecular analyses of the COI gene, based on intra- and interspecific genetic divergences (GDs), revealed that the borers were identified as two invasive species, Sphecodoptera sheni and Paranthrenella pinoakula sp. nov. The maximum intraspecific GD was found to be 1.9%, whereas the minimum interspecific GD was confirmed as 8.1%, indicating a distinct barcoding gap. Both the MJ network and NJ tree also showed that 18 haplotypes (Hs) were detected from the 52 COI sequences. The borers revealed a total of 17 Hs: (i) H1–H7 were detected in all seven regions with S. sheni; (ii) Wonju and Goyang populations of S. sheni revealed more than three Hs; (iii) H7 was closely connected with H8 of the Chinese population of S. sheni; (iv) H9–H10 were detected in other samples from the Wonju population with P. pinoakula sp. n. and were closely located with congeneric species. A maximum likelihood tree also revealed that P. pinoacula sp. n. nested within the congeneric species, genetically separating from S. sheni. Full article

Photuris female fireflies attract males of different firefly species by responding to their flashing signals; then, they try to capture and feed on them. This aggressive mimicry is considered a major selective pressure on the communication systems of the fireflies of the American continent. The intensity of this selective pressure is a function of its efficiency in prey capture. In this study, the rates of attraction and capture of males of the synchronous firefly Photinus palaciosi by the predatory females of Photuris lugubris are reported. Although the females attract numerous males, their hunting success is low. This result is consistent with the few previous measurements published. In agreement with the predicted coevolutionary race between predator and prey, behaviors consistent with predation avoidance in P. palaciosi and increasing prey encounters and prey deception by P. lugubris were observed. Full article

Trogoderma granarium Everts, the khapra beetle, is a serious pest of stored products throughout the world. Larvae pose a significant threat to stored products because they feed on >100 different commodities, possess the ability to enter facultative diapause, and are difficult to detect. Control methods for T. granarium include fumigation, contact insecticides, trapping, and insecticide-incorporated packaging. The objective of this study was to determine the residual efficacy of two insecticide formulations (methoprene + deltamethrin + piperonyl butoxide synergist Gravista^® and methoprene + deltamethrin, DiaconIGR^®Plus). These insecticides were evaluated on three stored product commodities, corn, wheat, and brown rice, by exposing T. granarium larvae during a 12-month testing period. Both formulations significantly reduced adult emergence on corn and wheat for 12 months and on brown rice for up to 6 months. Adult emergence was highest at month 12 for corn (8.41%), and brown rice (85.88%), and month 9 for wheat (39.52%), treated with DiaconIGR^®Plus or Gravista^®, respectively. A biological index used to measure the development of exposed larvae on the treated grain from the larval stage (low values) to adult emergence (high values) was lower (fewer adults) on corn and wheat compared to controls. Despite differences in formulations, each of these grain protectants could be utilized by stored commodity managers to protect commodities during storage and transportation for T. granarium when and if this pest is detected at USA ports of entry. Full article

Honey bee colonies have great societal and economic importance. The main challenge that beekeepers face is keeping bee colonies healthy under ever-changing environmental conditions. In the past two decades, beekeepers that manage colonies of Western honey bees (Apis mellifera) have become increasingly concerned by the presence of parasites and pathogens affecting the bees, the reduction in pollen and nectar availability, and the colonies’ exposure to pesticides, among others. Hence, beekeepers need to know the health condition of their colonies and how to keep them alive and thriving, which creates a need for a new holistic data collection method to harmonize the flow of information from various sources that can be linked at the colony level for different health determinants, such as bee colony, environmental, socioeconomic, and genetic statuses. For this purpose, we have developed and implemented the B-GOOD (Giving Beekeeping Guidance by computational-assisted Decision Making) project as a case study to categorize the colony’s health condition and find a Health Status Index (HSI). Using a 3-tier setup guided by work plans and standardized protocols, we have collected data from inside the colonies (amount of brood, disease load, honey harvest, etc.) and from their environment (floral resource availability). Most of the project’s data was automatically collected by the BEEP Base Sensor System. This continuous stream of data served as the basis to determine and validate an algorithm to calculate the HSI using machine learning. In this article, we share our insights on this holistic methodology and also highlight the importance of using a standardized data language to increase the compatibility between different current and future studies. We argue that the combined management of big data will be an essential building block in the development of targeted guidance for beekeepers and for the future of sustainable beekeeping. Full article

Varroa destructor is an ectoparasitic mite and is considered one of the most important causes of honey bee population loss. In the last years, substances of botanical origin have emerged as natural alternatives to diminish the mite population levels. Propolis is a natural product and is used by honey bees for multiple tasks, including protection from pathogens and parasites, and varroacidal activity of propolis extracts has been shown. In this study, we investigated the potential of propolis, collected by native Algerian honey bee subspecies (Apis mellifera intermissa and A. m. sahariensis) in different locations in Algeria and extracted by ultrasound, to control mites of V. destructor and tested the safety for the honey bees. The most important results were that the best propolis extracts at 10% killed 100% of the Varroa mites within 3–4 h in a Petri dish assay. In addition, when we sprayed A. m. intermissa bees infested with Varroa mites with a 10% concentration in a mini-hive setup, we scored a high mite mortality of 85–87% with the best propolis extracts, and importantly, there was no mortality in the bees. Our data demonstrated that propolis extracts in Algeria could be used in honey bee colonies by spraying against Varroa mite infestations, which may develop as an easy method for local beekeepers to control Varroa in their hives. Further research should investigate the mechanism of action. Full article

Understanding the factors associated with the species diversity and distribution of insect vectors is critically important for disease epidemiology. Black flies (Diptera: Simuliidae) are significant hematophagous insects, as many species are pests and vectors that transmit pathogens to humans and other animals. Ecological factors associated with black fly species distribution have been extensively examined for the immature stages but are far less well explored for the adult stage. In this study, we collected a total of 7706 adult black fly specimens from various locations in forests, villages and animal shelters in Thailand. The integration of morphology and DNA barcoding revealed 16 black fly taxa, including Simulium yvonneae, a species first found in Vietnam, which is a new record for Thailand. The most abundant species was the Simulium asakoae complex (n = 5739, 74%), followed by S. chumpornense Takaoka and Kuvangkadilok (n = 1232, 16%). The Simulium asakoae complex was dominant in forest (3786 of 4456; 85%) and village (1774 of 2077; 85%) habitats, while S. chumpornense predominated (857 of 1175; 73%) in animal shelter areas. The Simulium asakoae complex and S. nigrogilvum Summers, which are significant pests and vectors in Thailand, occurred at a wide range of elevations, although the latter species was found mainly in high (>1000 m) mountain areas. Simulium chumpornense, S. nodosum Puri and the S. siamense Takaoka and Suzuki complex occurred predominately in low (<800 m)-elevation areas. Simulium furvum Takaoka and Srisuka; S. phurueaense Tangkawanit, Wongpakam and Pramual; and S. nr. phurueaense were only found in high (>1000 m) mountain areas. A host blood meal analysis revealed that the S. asakoae; S. chamlongi Takaoka and Suzuki; S. nigrogilvum; S. chumpornense; and the S. striatum species group were biting humans. This is the first report of the latter two species biting humans. We also found that S. chumpornense was biting turkeys, and S. chamlongi was biting chickens, which are new host blood sources recorded for these species. In addition, we found that the S. feuerborni Edwards complex was biting water buffalo, which is the first report on the biting habits of this species. Full article

Tetranychus urticae is a highly polyphagous and global pest. Spider mites primarily feed on the underside of leaves, resulting in decreased photosynthesis, nutritional loss, and the development of chlorotic patches. We investigated the life tables of the two-spotted spider mite T. urticae on fungal endophyte Beauveria bassiana colonized and untreated plants of the common Phaseolus vulgaris L., a bean plant. Based on the age-stage, two-sex life table theory, data were evaluated. The mites raised on untreated plants had protonymphs, deutonymphs, and total pre-adult stage durations that were considerably shorter (1.76, 2.14, and 9.77 d, respectively) than the mites raised on plants that had been colonized (2.02, 2.45, and 10.49 d, respectively). The fecundity (F) varied from 28.01 eggs per female of colonized plants to 57.67 eggs per female of endophyte-untreated plants. The net reproductive rate (R₀) in the plants with and without endophytes was 19.26 and 42.53 brood, respectively. The untreated plants had an intrinsic rate of increase (rm) of 0.245 days as opposed to the colonized plants, which had an r of 0.196 days and a finite rate of increase (λ) (1.27 and 1.21, respectively). Population forecasts based on a two-sex, age-stage life table demonstrated the dynamism and variability of the stage structure. Furthermore, the colonization of B. bassiana had a negative impact on the growth and development of T. urticae. It lowered the adult mite life span, female fecundity, net reproduction rate, and intrinsic growth rate. We propose that future research should better use entomopathogenic fungal endophytes to understand host plant resistance strategies in integrated pest management. Full article

Silkworm ovary-derived BmN4 cells rely on chromatin-induced spindle assembly to form microtubule-based square mitotic spindles that ensure accurate segregation of holocentric chromosomes during cell division. The chromosome passenger protein Aurora B regulates chromosomal condensation and segregation, spindle assembly checkpoint activation, and cytokinesis; however, its role in holocentric organisms needs further clarification. This study examined the architecture and dynamics of spindle microtubules during prophase and metaphase in BmN4 cells and those with siRNA-mediated BmAurora B knockdown using immunofluorescence labeling. Anti-α-tubulin and anti-γ-tubulin antibodies revealed faint γ-tubulin signals colocalized with α-tubulin in early prophase during nuclear membrane rupture, which intensified as prophase progressed. At this stage, bright regions of α-tubulin around and on the nuclear membrane surrounding the chromatin suggested the start of microtubules assembling in the microtubule-organizing centers (MTOCs). In metaphase, fewer but larger γ-tubulin foci were detected on both sides of the chromosomes. This resulted in a distinctive multipolar square spindle with holocentric chromosomes aligned at the metaphase plate. siRNA-mediated BmAurora B knockdown significantly reduced the γ-tubulin foci during prophase, impacting microtubule nucleation and spindle structure in metaphase. Spatiotemporal BmAurora B expression analysis provided new insights into the regulation of this mitotic kinase in silkworm larval gonads during gametogenesis. Our results suggest that BmAurora B is crucial for the formation of multipolar square spindles in holocentric insects, possibly through the activation of γ-tubulin ring complexes in multiple centrosome-like MTOCs. Full article

Fireflies are a diverse group of bioluminescent beetles belonging to the family Lampyridae. Recent research on their diversity, evolution, behavior and conservation has greatly advanced our scientific understanding of these charismatic insects. In this review, we first summarize new discoveries about their taxonomic and ecological diversity, then focus on recent endeavors to identify and protect threatened fireflies around the world. We outline the main threats linked to recent population declines (habitat loss and degradation, light pollution, pesticide overuse, climate change and tourism) and describe relevant risk factors that predict which species will be particularly vulnerable to these threats. Although global coordination of firefly conservation efforts has begun only recently, considerable progress has already been made. We describe work by the IUCN SSC Firefly Specialist Group to identify species currently facing elevated extinction risks and to devise conservation strategies to protect them. To date, IUCN Red List assessments have been completed for 150 firefly taxa, about 20% of which face heightened extinction risks. The conservation status for many species has yet to be determined due to insufficient information, although targeted surveys and community science projects have contributed valuable new data. Finally, we highlight some examples of successful firefly habitat protection and restoration efforts, and we use the framework of the IUCN SSC Species Conservation Cycle to point out high-priority actions for future firefly conservation efforts. Full article

Mosquitoes, as disease vectors causing global morbidity and mortality through diseases like malaria, dengue, and Zika, necessitate mosquito population control methods. This study investigated the efficacy of nano-formulated insecticide-based sugar baits in controlling Anopheles gambiae populations and assessed their potential non-target impact on Coccinella septempunctata. This laboratory-based study employed thiolated polymer-coated attractive toxic sugar bait (ATSB) nano-formulations, delivering pesticides via nano-carriers. Adult and larvae populations of insects were collected from rice and cotton fields subjected to bioassays with 0.5% and 1% concentrations of each nano-formulated and conventional insecticide within ATSB solution, alongside a control 100% attractive sugar bait (ASB). Mosquitoes interacted overnight with insecticide-treated baits, and mortality was assessed. Further observations up to 72 h were conducted for potential delayed toxic effects. Results highlighted nano-ATSB carbosulfan’s effectiveness, particularly among organophosphates and pyrethroids. Among pyrethroids, nano-ATSB cypermethrin exhibited high efficacy, while Deltamethrin displayed lower mortality. Among organophosphates, nano-ATSB chlorpyrifos induced substantial mortality. The nano-formulations of insecticide were harmless against C. septempunctata compared to their conventional form. Nano-formulations demonstrated enhanced mortality rates and prolonged efficacy against mosquitoes, having a benign impact on non-target beetles. We expect these results to aid in developing effective plant protection products suitable for IPM practices. Full article

Naturally durable wood pre-dates preservative-treated wood and has been demonstrated to offer a suitable service life for certain applications where preservative-treated wood is not feasible. Heartwood extractives have been demonstrated to impart bio-deteriorative resistance to naturally durable wood species. These extractives are typically found in the heartwood of living trees and are produced either by the death of parenchyma cells or as the result of external stimuli. The mechanisms of natural durability are not well understood, as heartwood extractives can be extremely variable in their distribution, composition, and efficacy in both living and harvested trees. The underlying complexity of heartwood extractives has hindered their standardization in residential building codes for use as wood preservatives. The use of naturally durable lumber is not always feasible, as woods with exceptionally durable heartwood do not typically yield lumber with acceptable machining properties. A potential approach to overcome the inherent difficulty in establishing guidelines for the appropriate use of naturally durable wood is to focus solely on the extractives as a source of bioactive protectants based on the strategies used on living and dead wood to repel the agents of biodeterioration. This critical review summarizes the relevant literature on naturally durable woods, their extractives, and their potential use as bio-inspired wood protectants. An additional discussion will be aimed at underscoring the past difficulties in adopting this approach and how to overcome the future hurdles. Full article

(1) Background: The Gagr gene in Drosophila melanogaster’s genome originated from the molecular domestication of retrotransposons and retroviruses’ gag gene. In all Drosophila species, the Gagr protein homologs exhibit a conserved structure, indicative of a vital role. Previous studies have suggested a potential link between the Gagr gene function and stress responses. (2) Methods: We compared flies with Gagr gene knockdown in all tissues to control flies in physiological tests and RNA-sequencing experiments. (3) Results: Flies with the Gagr gene knockdown exhibited shorter lifespans compared to control flies. Transcriptome analysis revealed that Gagr knockdown flies showed elevated transcription levels of immune response genes. We used ammonium persulfate, a potent stress inducer, to elicit a stress response. In control flies, ammonium persulfate activated the Toll, JAK/STAT, and JNK/MAPK signaling pathways. In contrast, flies with the Gagr gene knockdown displayed reduced expression of stress response genes. Gene ontology enrichment analysis identified categories of genes upregulated under ammonium persulfate stress in control flies but not in Gagr knockdown flies. These genes are involved in developmental control, morphogenesis, and central nervous system function. (4) Conclusion: Our findings indicate the significance of the Gagr gene in maintaining immune response and homeostasis. Full article

The Varroa destructor mite infests Apis mellifera colonies and causes significant harm. Traditional treatments have become less effective because of mite resistance development and can also generate residues inside beehives. This study aimed to gauge the efficacy of a beehive-derived postbiotic in reducing V. destructor viability and to explore its synergies with organic compounds. Four lactic acid bacteria (LAB) species, Leuconostoc mesenteroides, Lactobacillus helsingborgensis, Bacillus velezensis, and Apilactobacillus kunkeei, were isolated and tested in a postbiotic form (preparations of inanimate microorganisms and/or their components) via bioassays. L. mesenteroides, L. helsingborgensis, and B. velezensis notably reduced the mite viability compared to the control, and they were further tested together as a single postbiotic product (POS). Further bioassays were performed to assess the impact of the POS and its combinations with oxalic acid and oregano essential oil. The simple products and combinations (POS/Oregano, POS/Oxalic, Oregano/Oxalic, and POS/Oregano/Oxalic) decreased the mite viability. The most effective were the oxalic acid combinations (POS/Oregano/Oxalic, Oxalic/Oregano, POS/Oxalic), showing significant improvements compared to the individual products. These findings highlight the potential of combining organic products as a vital strategy for controlling V. destructor infection. This study suggests that these combinations could serve as essential tools for combating the impact of mites on bee colonies. Full article

The endangered terrestrial firefly Pyrocoelia pectoralis (Olivier) is endemic to China. Populations of P. pectoralis have decreased dramatically due to urbanization and pollution. Breeding and re-introduction to a suitable habitat may save the species from becoming extinct. Because of its polyandrous character, an investigation into the possibility of sperm competition and paternity outcomes from multiple matings was initiated to better understand its reproductive physiology. To achieve these goals, 13 SSR markers were developed. The results of paternity experiments indicate there is a significant difference between P3 and P1 or P2. The female reproductive system has three spermathecae which accept sperm from different matings, and no bursa or spermatophore-digesting organ is developed. Our research established that multiple inseminations with sperm from different males occur, leading to competition between ejaculates. The benefits of such competition include an increasing number of sperm in the ejaculates of competing males and the consequential increase in fertilized eggs (thus, fecundity), and thereby a higher chance of genetic diversity and fitness in the offspring of the firefly P. pectoralis. Full article

Understanding the fundamental life cycle and reproductive behavior of a pest insect is essential for developing efficient control strategies; however, much of this knowledge remains elusive for a multitude of insects, including the cotton seed bug, Oxycarenus hyalinipennis. Here, we report the results of our comprehensive study on the cotton seed bug’s life cycle, including mating behavior, adult lifespan, and egg-to-adulthood development. Our findings showed that adult males and females began mating as early as three days after emerging (75%), and the frequency of mating increased to 100% by the fifth day. Mated females commenced oviposition on cotton seeds as early as two days after mating, with a cumulative mean number of 151 fertile eggs oviposited during the first oviposition cycle. Furthermore, around 10% of eggs from both mated and unmated females remained unfertilized. The first instar nymphs began emerging approximately seven days following oviposition. To track their development, we monitored the newly hatched nymphs daily until they reached adulthood. There were five nymphal stages, which cumulatively took roughly 28 to 30 days. Notably, mating positively influenced the survivorship and lifespan of adult O. hyalinipennis. Mated males and females exhibited median lifespans of 28 and 25 days, respectively. In contrast, unmated males and females only lived for a median lifespan of 9.5 days, about one-third that of the mated O. hyalinipennis. Our study provides key insights into the O. hyalinipennis life history for new IPM strategies. Full article

The level of diversity and abundance of darkling beetles (Coleoptera, Tenebrionidae) is the main difference between the late Pleistocene and modern insect faunas of arid regions. In the Pleistocene assemblages they are extremely rare, whereas in the modern ones they predominate. It is assumed that the reason for their rarity in fossil entomological complexes is their lack of cold resistance. The supercooling points (SCP) and low lethal temperatures (LLT) of adults from five species of Altai darkling beetles that overwinter in the soil and larvae from one such species were measured in the laboratory. All beetles supercooled at negative temperatures but could not survive freezing, with the average SCP of the most cold-resistant species between −25.7 and −21.7 °C (Bioramix picipes, Anatolica dashidorzsi, and Penthicus altaicus). However, 50% of the individuals from different species in the experiment died after exposure during two days at temperatures ranging from −22 to −20 °C. The focal species are distributed in parts of Central Asia with an extreme continental climate, and the temperatures measured in the soil of these natural areas turned out to be lower than or close to the limit of cold resistance of the beetles. Overwintering of darkling beetles is therefore only possible in areas with deep snow: in hollows, under bushes, and under large cereals. Darkling beetles with poor cold resistance could not have existed in the colder climate of the late Pleistocene, which explains their absence from fossil fauna. Full article

The molecular mechanisms underlying insect gall formation remain unclear. A major reason for the inability to identify the responsible genes is that only a few systems can be experimentally validated in the laboratory. To overcome these problems, we established a new galling insect model, Smicronyx madaranus. Our manipulation experiments using nail polish sealing and insecticide treatment revealed an age-dependent change in gall formation by S. madaranus; adult females and larvae are responsible for gall induction and enlargement, respectively. Furthermore, it has been suggested that substances released during oviposition and larval feeding are involved in each process. Phylogenetic analysis showed that gall-forming weevils, including S. madaranus, belong to two distinct lineages that utilize different host plants. This may indicate that gall-forming traits evolved independently in these Smicronyx lineages. The efficacy of RNA interference (RNAi) in S. madaranus was confirmed by targeting the multicopper oxidase 2 gene. It is expected that the mechanisms of gall formation will be elucidated by a comprehensive functional analysis of candidate genes using RNAi and the S. madaranus galling system in the near future. Full article

The African citrus psyllid, Trioza erytreae, is one of the two vectors of Huanglongbing, the most serious citrus disease worldwide. The first detection of T. erytreae in the European mainland was on the northwest of the Iberian Peninsula in 2014. Since then, the pest has spread throughout northern Spain (Galicia, Asturias, Cantabria, País Vasco) and along the western Atlantic coast of Portugal (from the Douro e Minho region to the Algarve). We conducted a series of laboratory experiments on lemon plants at different temperatures (from 8 to 34 °C) and humidity conditions (from 40 to 90%) to find out the influence of extreme temperatures and relative humidities (RHs) on the mortality, development and reproduction of T. erytreae. Our results show that temperatures above 30 °C and below 10 °C are very detrimental for nymphal development and nymphs were unable to reach the adult stage. Furthermore, eggs were unable to hatch under temperatures above 33 °C and below 8 °C. Adult mortality was highest at 34 °C and killed more than 50% of the population. We also found that relative humidity is crucial for the development and survival of T. erytreae. Nymphs were unable to reach the adult stage at an RH of 90% and 40%. Also, fecundity was significantly reduced at 90 and 40% RH, and fertility was lowest at 40% RH. Nymphal mortality was highest at an RH of 40%, which was the most detrimental humidity among all tested for the survival and development of T. erytreae. Our work concludes that T. erytreae establishment and spread will be maximum in regions with a temperate and humid climate, being rare in regions where dry and hot weather conditions predominate. Full article

Understanding the seasonal dynamics inherent to non-crop host–fruit fly–parasitoid interactions is vitally important for implementing eco-friendly pest control strategies. This study assessed the abundance and seasonal infestation levels of three pest fly species, Ceratitis capitata (Wiedemann), Anastrepha fraterculus (Wiedemann), Drosophila suzukii (Matsumura), as well as the related saprophytic drosophilids, and their natural parasitism in a disturbed wild habitat characterized by non-crop hosts in northwestern Argentina over 40 months. Juglans australis Griseb (walnut), Citrus aurantium L. (sour orange), Eriobotrya japonica (Thunb.) Lindley (loquat), Prunus persica (L.) Batsch (peach), and Psydium guajava L. (guava) were sampled throughout their fruiting seasons. Fruits were collected from both the tree canopies and the ground. The most abundant puparia was A. fraterculus, followed by C. capitata and D. suzukii. Drosophila species from the D. melanogaster group were highly abundant only in fallen fruits. Spatiotemporal overlaps of different host fruit availability provided suitable sources for pest proliferation throughout the year. The populations of both invasive pests peaked from December to January, and were related to the highest ripe peach availability, whereas the A. fraterculus population peaked from February to April, overlapping with the guava fruiting period. The three pest fly species were parasitized mainly by three generalist resident parasitoids, which are potential biocontrol agents to use within an integrated pest management approach. Full article

Certain soil insects, such as the root-damaging larvae of the maize pest Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), are increasingly difficult to control because of recent bans of some insecticides. An alternative and safer approach may be the development of biopesticides based on entomotoxic defense proteins of higher fungi. Many of these potentially interesting proteins are protease inhibitors, and some have been shown to adversely affect insects. We examined the effects of the cysteine protease inhibitors macrocypin 1, 3, and 4 from Macrolepiota procera, clitocypin from Clitocybe nebularis, and cocaprin 1 and the serine protease inhibitor cospin 1 from Coprinopsis cinerea on D. v. virgifera. We confirmed the inhibition by mycocypins of the cysteine catalytic-type proteolytic activities in gut extracts of larvae and adults. The inhibition of pGlu-Phe-Leu-hydrolyzing activity was stronger than that of Z-Phe-Arg-hydrolyzing activity. Mycocypins and cospin resisted long-term proteolytic digestion, whereas cocaprin 1 was digested. Bioassays with overlaid artificial diet revealed no effects of proteins on neonatal mortality or stunting, and no effects on adult mortality. Immersion of eggs in protein solutions had little effect on egg hatching or mortality of hatching neonates. Microscopic analysis of the peritrophic matrix and apical surface of the midguts revealed the similarity between larvae of D. v. virgifera and the chrysomelid Leptinotarsa decemlineata, which are sensitive to these inhibitors. The resistance of D. v. virgifera to fungal protease inhibitors is likely due to effective adaptation of digestive enzyme expression to dietary protease inhibitors. We continue to study unique protein complexes of higher fungi for the development of new approaches to pest control. Full article

Nosema ceranae (N. ceranae) infection is prevalent globally, causing a decline in bee populations and significant economic losses to apiarists. Although several methods have been proposed for diagnosing Nosema infections, limitations in these methods have hindered their broad applications. Therefore, this current study aimed to develop a specialized method for diagnosing Nosema infections. To achieve this, a sandwich enzyme-linked immunosorbent assay (ELISA) and immunochromatography assay (ICG) were developed, and their effectiveness in screening and diagnosing Nosema infection was assessed. In sandwich ELISA, the combination of the monoclonal antibodies (mAb) 19B2 and biotinylated-19B2 exhibited stronger binding affinity to the antigen than did other combinations of mAbs that were tested. Furthermore, the antigen detection limit achieved with the sandwich ELISA surpassed that previously reported with Western blotting. The ICG was designed using the same antibody combination as that used in sandwich ELISA; however, the assay exhibited a lower diagnostic ability for Nosema infection than the ELISA. The diagnostic models developed in this study offer practical applications for conducting rapid nosemosis detection tests. These innovative techniques will help to improve the timely identification and management of nosemosis. Full article

The Asian citrus psyllid (ACP) is a citrus pest and insect vector of “Candidatus Liberibacter asiaticus”, the causal agent of citrus greening disease. Double-stranded RNA (dsRNA) biopesticides that trigger RNA interference (RNAi) offer an alternative to traditional insecticides. Standardized laboratory screening of dsRNA requires establishing the minimal effective concentration(s) that result in effective RNAi “penetrance” and trigger RNAi, resulting in one or more measurable phenotypes, herein, significant gene knockdown and the potential for mortality. In this study, knockdown was evaluated for a range of dsRNA concentrations of three ACP candidate genes, clathrin heavy chain (CHC), vacuolar ATPase subunit A (vATPase-A), and sucrose non-fermenting protein 7 (Snf7). Gene knockdown was quantified for ACP teneral adults and 3rd instar nymphs allowed a 48 h ingestion-access period (IAP) on 10, 50,100, 200, and 500 ng/µL dsRNA dissolved in 20% sucrose followed by a 5-day post-IAP on orange jasmine shoots. Significant gene knockdown (p < 0.05) in ACP third instar nymphs and adults ranged from 12–34% and 18–39%, 5 days post-IAP on dsRNA at 10–500 and 100–500 ng/µL, respectively. The threshold concentration beyond which no significant gene knockdown and adult mortality was observed post-48 h IAP and 10-day IAP, respectively, was determined as 200 ng/µL, a concentration indicative of optimal RNAi penetrance. Full article

The phylogenetic status of the family Nitidulidae and its sister group relationship remain controversial. Also, the status of the subfamily Meligethinae is not fully understood, and previous studies have been mainly based on morphology, molecular fragments, and biological habits, rather than the analysis of the complete mitochondrial genome. Up to now, there has been no complete mitochondrial genome report of Meligethinae. In this study, the complete mitochondrial genomes of Meligethinus tschungseni and Brassicogethes affinis (both from China) were provided, and they were compared with the existing complete mitochondrial genomes of Nitidulidae. The phylogenetic analysis among 20 species of Coleoptera was reconstructed via PhyloBayes analysis and Maximum likelihood (ML) analysis, respectively. The results showed that the full lengths of Meligethinus tschungseni and Brassicogethes affinis were 15,783 bp and 16,622 bp, and the AT contents were 77% and 76.7%, respectively. Each complete mitochondrial genome contains 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and a control region (A + T-rich region). All the PCGs begin with the standard start codon ATN (ATA, ATT, ATG, ATC). All the PCGs terminate with a complete terminal codon, TAA or TAG, except cox1, cox2, nad4, and nad5, which terminate with a single T. Furthermore, all the tRNAs have a typical clover-leaf secondary structure except trnS1, whose DHU arm is missing in both species. The two newly sequenced species have different numbers and lengths of tandem repeat regions in their control regions. Based on the genetic distance and Ka/Ks analysis, nad6 showed a higher variability and faster evolutionary rate. Based on the available complete mitochondrial genomes, the results showed that the four subfamilies (Nitidulinae, Meligethinae, Carpophilinae, Epuraeinae) of Nitidulidae formed a monophyletic group and further supported the sister group relationship of Nitidulidae + Kateretidae. In addition, the taxonomic status of Meligethinae and the sister group relationship between Meligethinae and Nitidulinae (the latter as currently circumscribed) were also preliminarily explored. Full article

The application of the sterile insect technique (SIT) requires the adaptation of insects to mass-rearing conditions. It is generally accepted that this adaptation may include a reduction in genetic diversity and an associated loss of desirable characteristics for the effective performance of sterile insects in the field. Here, we compare the genetic diversity of two mass-reared strains of the Mexican fruit fly, Anastrepha ludens, and a wild (WIL) population collected near Tapachula, Mexico, using seven DNA microsatellites as molecular genetic markers. The mass-reared strains were a bisexual laboratory strain (LAB) with approximately 130 generations under mass-rearing and a genetic sexing strain, Tapachula-7 (TA7), also under mass-rearing for 100 generations. Our results revealed an overall low level of genetic differentiation (approximately 15%) among the three strains, with the LAB and WIL populations being genetically most similar and TA7 most genetically differentiated. Although there were some differences in allele frequencies between strains, our results show that overall, the adaptation to mass-rearing conditions did not reduce genetic variability compared to the wild sample in terms of heterozygosity or allelic richness, nor did it appear to alter the level of inbreeding with respect to the wild populations. These results are contrary to the general idea that mass-rearing always results in a reduction in genetic diversity. Overall, our findings can contribute to a better understanding of the impact that adaptation to mass-rearing conditions may have on the genetic make-up of strains. Full article

Biogeographically, Anatolia harbours a rich diversity of short-horned grasshoppers (Orthoptera, Caelifera). The number of species recorded from Anatolia so far stands at 300. They inhabit diverse habitats ranging from arid Eremial to Euro-Siberian-like montane meadows, aligning with the topographical and climatological heterogeneity of Anatolia. Alongside some swarming species, the pest potential of several pullulating species needs attention. This is especially important concerning global warming, a scenario expected to be more severe in the Northern Mediterranean Basin in general and Anatolia specifically. A faunal list of biogeographic Anatolia, the area extending from the Aegean Sea in the west to the intermountain basin of the Caucasus in the northeast, the lowlands of Lake Urmia in the east, and Mesopotamia in the southeast, was developed. The recorded species were classified according to the phytogeographical provinces of Anatolia. Distributions of the species with the potential for pullulating were modelled using ecological-niche-modelling approaches for the present and future. The results have the potential to lead to the development of a concept that merges biogeography and the pest potential of certain Anatolian grasshopper species. Our results reveal the following: (i) Acrididae and Pamphagidae are the most diverse families represented in Anatolia; (ii) roughly 40% of Caelifera and 71% of Pamphagidae are endemics, suggesting Anatolia is a biodiversity hotspot; (iii) according to Caelifera diversity, the phytogeographical provinces of Anatolia follow an order of Irano-Anatolia, Euro-Siberia, Mediterranean, and Mesopotamia; and (iv) based on ecological modelling and personal observations, Dociostaurus maroccanus, Locusta migratoria, Calliptamus italicus, Heteracris pterosticha, Notostaurus anatolicus, Oedipoda miniata, and O. schochii should be monitored regarding their pest potential. Full article

Toxicological studies have shown that the American cockroach Periplaneta americana (Linnaeus) is a classical model for studying the mode of action of commonly used insecticides. In a previous study, we demonstrated that thiamethoxam and clothianidin decreased locomotor activity in an open-field-like apparatus. Here, we tested the effect of the neonicotinoid acetamiprid when applied orally, topically, or injected into the haemolymph. We found that acetamiprid was also able to impair locomotor activity in the open-field-like apparatus. When treated with acetamiprid, a strong alteration in locomotor activity was observed 1 h, 24 h, and 48 h after haemolymph and topical applications. Oral application induced an impairment of locomotor activity at 24 h and 48 h. A comparison of the present data with our previously published results showed that neonicotinoids were more active when injected into the haemolymph compared to oral and topical applications. These findings increased our understanding of the effect of neonicotinoid insecticides on insect locomotor activity, and demonstrated that the cyano-substituted neonicotinoid, acetamiprid, was able to alter cockroach locomotor activity. Full article

Predator–prey coevolution, particularly chemo-ecological arms races, is challenging to study as it requires the integration of behavioral, chemical ecology, and phylogenetic studies in an amenable system. Moths of the genus Hemileuca (Saturniidae) are colorful, diurnal, and fast and often fly well above the vegetation canopy layer. However, several Hemileuca species have been reported as being captured in spider webs, specifically Argiope species (Araneidae). Female Hemileuca are known to produce mating pheromones and spiders of the Araneidae family are known to use pheromone lures to attract lepidopteran prey. We presented primarily female Argiope aurantia, which are attractive to male Anisota pellucida (Saturniidae), to different populations of Hemileuca species across the southern and western United States to categorize the homing response strength of different species of male Hemileuca. When we mapped these Argiope lure attraction strength categories onto the most recently published Hemileuca phylogeny, the behavioral patterns suggested a potential co-evolutionary arms race between predators and prey. Males of Hemileuca maia, H. grotei, and H. nevadensis (all in the same clade) appeared to have no attraction to A. aurantia, while H. magnifica and H. hera (within a different, separate clade) appeared to be strongly attracted to A. aurantia, but H. nuttalli (also within the H. hera and H. magnifica clade) displayed no attraction. Furthermore, Hemileuca eglanterina (yet a different clade) displayed strong, weak, and no attraction to A. aurantia, depending on the population. These apparent clade partitioning patterns of Argiope lure effectiveness and within-species variation in Hemileuca lure responses suggest a predator–prey coevolutionary history of measures and countermeasures. Full article

In strawberry production, western flower thrips (WFT) and two-spotted spider mites (TSSM) inflict feeding damage and reduce the yield. Biological control for these pests often includes phytoseiid predatory mites and entomopathogenic fungi. The hemipteran family Nabidae have been reported as prominent predators in open-field strawberry. Nabis americoferus Carayon is a new biocontrol agent developed in Canada. This study examined if this species was a good candidate for integration with biological control for greenhouse strawberry production. The laboratory trials showed that Phytoseiulus persimilis Athias-Henriot and Amblyseius swirskii Athias-Henriot were compatible with N. americoferus, especially when alternative food was available. In contrast, the nabid was not compatible with the Beauveria bassiana (Balsamo) GHA strain. A greenhouse cage study was conducted to determine if it was beneficial to add N. americoferus to the phytoseiid-mites-based biological control program for WFT and TSSM in greenhouse strawberry. The release of N. americoferus on a banker plant together with the placement of sachets of Neoseiulus cucumeris (Oudemans) and Neoseiulus californicus (McGregor) was beneficial, not only potentially reducing the number of sachet applications, but also providing better pest control than phytoseiid mites alone. Neither the phytoseiids nor the N. americoferus numbers were significantly affected by the presence of each other. Full article

Chironomids are found in all types of freshwater habitats; they are a ubiquitous and highly diverse group of aquatic insects. Plitvice Lakes National Park is the oldest and largest national park in Croatia and consists of numerous and diverse freshwater habitats, making the area an ideal location for long-term research into the chironomid emergence patterns and phenology. The main objectives of this study were to identify the composition of the chironomid community, determine the phenology of the identified species, and assess the main factors influencing their emergence in Plitvice Lakes. During 14 years of research, more than 13,000 chironomids belonging to more than 80 species were recorded. The most abundant species was found to be Parametriocnemus stylatus. The highest abundance of chironomids was recorded in lotic habitats with faster water current over substrates of moss and algae and pebbles. Water temperature and the availability of organic matter were found to be the main factors that drive chironomid emergence at the tufa barrier studied. In the last years of this study, a prolonged flight period was observed. Although this is not statistically significant (at this stage of the study), it could be due to a higher water temperature in winter. Full article

Monolepta is one of the diverse genera in the subfamily Galerucinae, including 708 species and 6 sub-species worldwide. To explore the information on the mitogenome characteristics and phylogeny of the section “Monoleptites”, especially the genus Monolepta, we obtained the newly completed mitochondrial genomes (mitogenomes) of four Monolepta species using high-throughput sequencing technology. The lengths of these four new mitochondrial genomes are 16,672 bp, 16,965 bp, 16,012 bp, and 15,866 bp in size, respectively. All four mitochondrial genomes include 22 transfer RNA genes (tRNAs), 13 protein-coding genes (PCGs), 2 ribosomal RNA genes (rRNAs), and one control region, which is consistent with other Coleoptera. The results of the nonsynonymous with synonymous substitution rates showed that ND6 had the highest evolution rate, while COI displayed the lowest evolution rate. The substitution saturation of three datasets (13 PCGs_codon1, 13 PCGs_codon2, 13 PCGs_codon3) showed that there was no saturation across all datasets. Phylogenetic analyses based on three datasets (ND1, 15 genes of mitogenomes, and 13 PCGs_AA) were carried out using maximum likelihood (ML) and Bayesian inference (BI) methods. The results showed that mitogenomes had a greater capacity to resolve the main clades than the ND1 gene at the suprageneric and species levels. The section “Monoleptites” was proven to be a monophyletic group, while Monolepta was a non-monophyletic group. Based on ND1 data, the newly sequenced species whose antennal segment 2 was shorter than 3 were split into several clades, while, based on the mitogenomic dataset, the four newly sequenced species had close relationships with Paleosepharia. The species whose antennal segment 2 was as long as 3 were split into two clades, which indicated that the characteristic of “antennal segment 2 as long as 3” of the true “Monolepta” evolved multiple times in several subgroups. Therefore, to explore the relationships among the true Monolepta, the most important thing is to perform a thorough revision of Monolepta and related genera in the future. Full article