Search Results (300)

Fruit flies that belong to the genus Zeugodacus (Diptera: Tephritidae) pose significant threats as invasive pests of agricultural crops in Asia and sub-Saharan Africa. The intensification of transboundary trade in fresh horticultural produce has increased the risk of introducing invasive species such as fruit flies, more so through the inadvertent transport of their immature developmental stages. Such immature stages of fruit flies belonging to the Tephritidae family are frequently intercepted at the international borders worldwide and are unable to be identified to the species level using morphological characteristics. Molecular identification using mitochondrial Cytochrome Oxidase I (COI) gene has proven to be quite useful, as they are not constrained by developmental stages, sex, or colour morphs of the pest species in question. Also, real-time PCR-based species-specific assays offer quicker turnaround time since they do not require any post-PCR procedures. This study evaluated the utility of a real-time PCR assay based on the COI gene region to identify Zeugodacus cucurbitae from other Tephritid species. The developed real-time PCR assay provides a swift and precise way of discriminating between these highly invasive pest species during an interception event for rapid decision making. High specificity, having no cross-reactions with closely related Tephritids, and sensitivity of the developed assay will be extremely useful in discriminating Z. cucurbitae from other closely related fruit fly species. Z. cucurbitae-specific real-time PCR developed in this study is appropriate for organizations that carry out routine diagnostics to facilitate fresh produce imports and exports. Our assay is fully optimized for rapid deployment at international borders, offering reliable detection of the target species regardless of developmental stage, sex, or geographic origins. Full article

Broad bean is one of the most important leguminous crops worldwide. However, its productivity is greatly affected by the infestation of Aphis fabae and Aphis craccivora (Hemiptera: Aphididae). The main objective of the current study was to identify the most susceptible phenological stages of the broad bean variety (Histal) against black aphids’ herbivory. This had been achieved through an evaluation of plant resistance mechanisms such as antixenosis and antibiosis. The results regarding an antixenosis test revealed that the four tested phenological stages of V. faba did not have a significant effect on the preference of A. craccivora and A. fabae towards the crop plant. Overall, a slightly higher number of adults settled on the three and four unfolded leaves’ stage of the crop plant. Similarly, the highest number of developed embryos were found in the four leaves’ stage of the crop, and the lowest in the second leaf stage. The adult body size of A. craccivora was slightly larger in the case of the three unfolded leaves. Furthermore, the maximum body size of A. fabae adults was recorded in the case of the first unfolded leaf stage crop. Linear correlations between the biological parameters for both species revealed only one significant relationship between developed and total embryos for A. craccivora. The results of the current study highlight the need to protect broad bean crops against infestations of black aphids, i.e., A. craccivora and A. fabae. This is essential for reducing direct damage and preventing the transmission of phytoviruses. However, future studies should aim to evaluate the susceptibility of all developmental phenological stages of the crop against black aphids to mitigate potential crop losses. Full article

Antipredator behaviors in animals often vary with developmental stage, microhabitat, and social context, yet few studies examine how these factors interact in species that undergo ontogenetic shifts in chemical defense. The spotted lanternfly (Lycorma delicatula) is an invasive planthopper whose nymphs transition from cryptically colored early instars to aposematically colored fourth instars that feed primarily on chemically defended host plants. We conducted 1460 simulated predator attacks on nymphs across four developmental stages to examine how antipredator behavior varies with instar, plant location (leaf vs. stem), host plant species, and local conspecific density. Nymphs exhibited three primary responses: hiding, sidestepping, or jumping. We found that location on the plant had the strongest effect, with nymphs on stems more likely to hide than those on leaves. Older instars were significantly less likely to hide and more likely to sidestep, particularly on stems, suggesting reduced reliance on energetically costly escape behaviors as chemical defenses accumulate. First instars were less likely to jump from their preferred host plant (tree of heaven) compared to other plant species. Higher local conspecific density reduced hiding probability, likely due to the dilution effect. These results demonstrate that antipredator strategies in L. delicatula are flexibly deployed based on developmental stage, microhabitat structure, and social context, with implications for understanding evolution of antipredator behavior in chemically protected species. Full article

Honey bees are essential pollinators for the ecosystem and food crops. However, their health and survival face threats from both biotic and abiotic stresses. Fungi, microsporidia, and bacteria might significantly contribute to colony losses. Therefore, rapid and sensitive diagnostic tools are crucial for effective disease management. In this study, molecular assays were developed to quickly and efficiently detect the main honey bee pathogens: Nosema ceranae, Aspergillus flavus, Paenibacillus larvae, and Black queen cell virus. In this context, new primer pairs were designed for use in quantitative Real-time PCR (qPCR) reactions. Various protocols for extracting total nucleic acids from bee tissues were tested, indicating a CTAB-based protocol as the most efficient and cost-effective. Furthermore, excluding the head of the bee from the extraction, better results were obtained in terms of quantity and purity of extracted nucleic acids. These assays showed high specificity and sensitivity, detecting up to 250 fg of N. ceranae, 25 fg of P. larvae, and 2.5 pg of A. flavus DNA, and 5 pg of BQCV cDNA, without interference from bee DNA. These qPCR assays allowed pathogen detection within 3 h and at early stages of infection, supporting timely and efficient management interventions. Full article

Dysbiosis is a strategy to control insect pests through disrupting symbiotic bacteria essential for their life cycle. The olive fly, Bactrocera oleae, has been considered a suitable system for dysbiosis, as the insect is strictly dependent on its unique symbiont Candidatus Erwinia dacicola. Here, we review older and recent results from studies of the interaction of the symbiont and its host fly. We then discuss possible methods for disrupting the symbiosis as a means to control the fly. Specifically, we summarize studies using microscopy methods that have investigated in great detail the organs where the bacterium resides and it is always extracellular. Furthermore, we discuss how genome sequences of both host and bacterium can provide valuable resources for understanding the interaction and transcriptomic analyses that have revealed important insights that can be exploited for dysbiosis strategies. We also assess experiments where compounds have been tested against the symbiont. The hitherto limited efficacy in decreasing bacterial abundance suggests that novel molecules and/or new ways for the delivery of agents will be important for successful dysbiosis strategies. Finally, we discuss how gene drive methods could be implemented in olive fly control, though a number of hurdles would need to be overcome. Full article

Tetranychus urticae, commonly known as the two-spotted spider mite, is a highly adaptable and polyphagous arthropod in the family Tetranychidae, capable of feeding on over 1200 plant species, including strawberries (Fragaria × ananassa Duch.). The fitness and microbiota of herbivorous arthropods can vary significantly across different plant species and cultivars. In this study, we investigated the fecundity, longevity, growth rate, and microbiota composition of T. urticae reared on seven Chinese strawberry cultivars: Hongyan (HY), Yuexiu (YX), Tianshi (TS), Ningyu (NY), Xuetu (XT), Zhangjj (ZJ), and Xuelixiang (XLX). Our findings revealed significant differences among cultivars: mites reared on the XT cultivar exhibited the highest fecundity (166.56 ± 7.82 eggs), while those on XLX had the shortest pre-adult period (7.71 ± 0.13 days). Longevity was significantly extended in mites reared on XLX, XT, and NY cultivars (25.95–26.83 days). Microbiota analysis via 16S rRNA sequencing showed that Proteobacteria dominated (>89.96% abundance) across all mite groups, with Wolbachia as the predominant symbiont (89.58–99.19%). Male mites exhibited higher bacterial diversity (Shannon and Chao1 indices) than females, though Wolbachia abundance did not differ significantly between sexes or cultivars. Functional predictions highlighted roles of microbiota in biosynthesis, detoxification, and energy metabolism. These findings underscore the influence of host plant variety on T. urticae fitness and microbiota composition, suggesting potential strategies for breeding resistant strawberry cultivars and leveraging microbial interactions for pest management. Full article

This study assessed the host plant selection behavior of female stalk-eyed flies (SEFs) or Diopsis apicalis, where a Y-tube olfactometer was used to compare SEF attraction to the odor of leaves from four rice varieties (ITA306, WAB56-104, CG14, and RAM55). Another step of the evaluation consisted of pairing leaf odors from two rice varieties. Also, potted plants of the tested varieties were displayed in a screened cage and submitted to female SEF selection. The results indicated that the odor produced by leaves from rice varieties CG14, WAB56-104, and ITA306 significantly attracted SEFs, at rates of 81%, 70%, and 97%, respectively, while SEF females were rarely attracted by the odor of leaves from the resistant rice variety RAM55, at a rate of 35%. The results suggested that the use of a Y-tube olfactometer was similar to the use of a screened cage. The resistance exhibited by rice variety CG14 against SEFs is related to an antibiosis interaction acting as bait, while that in RAM55 is an antixenosis one. Farmers can plant the traditional CG14 variety on the edge of rice fields to draw SEFs and poison their larvae. However, RAM55 can be inserted in an intercropping system to repel SEFs from laying eggs. The authors recommend CG14 and RAM55 as candidates for breeding to create resistant lines against SEF. Full article

Insects are often associated with diverse microorganisms that enhance their metabolism and nutrient assimilation. These microorganisms, residing in the insect’s gut, play a crucial role in breaking down complex molecules into simpler compounds essential for the host’s growth. This study investigates the diversity and functional potential of symbiotic bacteria in the gut of Arsenura armida (Lepidoptera: Saturniidae) larvae, an edible insect from southeastern Mexico, using culture-dependent and metagenomic approaches. Bacterial strains were isolated from different gut sections (foregut, midgut, and hindgut) and cultured on general-purpose media. Isolates were identified through 16S rRNA gene sequencing and genomic fingerprinting. Metagenomics revealed the bacterial community structure and diversity, along with their functional potential. A total of 96 bacterial strains were isolated, predominantly Gram-negative bacilli. Rapidly growing colonies exhibited enzymatic activity, cellulose degradation, and sugar production. Phylogenetic analysis identified eight genera, including Acinetobacter, Bacillus, Enterobacter, Pseudomonas, and others, with significant cellulose-degrading capabilities. Metagenomics confirmed Bacillota as the most abundant phylum. These complementary methods revealed abundant symbiotic bacteria with key metabolic roles in A. armida, offering promising biotechnological applications in enzymatic bioconversion and cellulose degradation. Full article

Insects and their endosymbionts have a close mutualistic relationship. However, the precise nature of the bacterial endosymbiont-mediated interaction between host plants and whitefly Bemisia tabaci MEAM1 is still unclear. In the present study, six populations of Bemisia tabaci MEAM1 sharing the same genetic background were established by rearing insects for ten generations on different host plants, including poinsettia, cabbage, cotton, tomato, and tobacco, and an additional population was reared on cotton and treated with antibiotics. The physiological and nutritional traits of the insects were found to be dependent on the host plant on which they had been reared. Systematic analysis was conducted on the endosymbiont titers, the amino acid molecules and contents, as well as developmental and oviposition changes in the MEAM1 populations reared on each host plant tested. The results indicate that B. tabaci contained the primary symbiont Portiera and the secondary symbionts Hamiltonella and Rickettsia. In addition, the titer of endosymbiotic bacteria in females is higher than that in males. Among the MEAM1 populations reared on each host plant, the variation pattern of Portiera titer generally corresponded with changes in biological characteristics (body length, weight and fecundity) and AA contents. This suggests that changes in the amino acid contents and biological characteristics of different B. tabaci populations may be due to changes in the Portiera content and the differences in the nutrition of the host plants themselves. Our findings were further confirmed by the reduction in Portiera with antibiotic treatment. The amino acids, body size, body weight, and fecundity of B. tabaci were all reduced with the decrease in the Portiera titer after antibiotic treatment. In summary, our research revealed that host plants can affect the content of symbiotic bacteria, particularly Portiera, and subsequently affect the nutrition (i.e., the essential amino acids content) of host insects, thus changing their biological characteristics. Full article

Hermetia illucens, the black soldier fly, is a common and widespread fly of the family Stratiomyidae. Its ability to grow on contaminated substrates suggests the production of antimicrobial peptides that enable its survival. This study aimed to verify the impact of direct and indirect infection with Salmonella enteritidis on the expression of defensins and cecropins in Hermetia illucens larvae. In addition to an infection with a microorganism, it was interesting to verify if the expression of peptides and the relative action of hemolymph changed in larvae subjected to mechanical stress by abdominal puncture. The peptide fraction of the hemolymph of infected larvae was tested using antibiogram and minimum inhibitory concentration tests against Salmonella enteritidis and Salmonella typhimurium. Both molecular and microbiological tests were carried out at three different time points, on larvae not subjected to any treatment (T-0), four hours after treatment (T-1), and 24 h after treatment (T-2). The results of the microbiological tests showed the antimicrobial action of the peptide fraction of the hemolymph against both S. typhimurium and S. enteritidis; for the latter one, the action was more marked. Interesting results were also found for larvae subjected only to mechanical stress by puncture. Molecular tests on the expression of defensins and cecropins were in full agreement with those obtained in the microbiological tests, with expression more pronounced in larvae infected directly with Salmonella enteritidis. Temporal and condition-specific regulation of defensins and cecropins highlights the complexity of the immune response and suggests sophisticated mechanisms by which the host fine-tunes antimicrobial peptide expression to enhance pathogen defense while preventing excessive immune activation. Full article

(1) Background: The larvae of Dioryctria sylvestrella typically bore into the shoots and cones of Pinus koraiensis, increasing tree breakage risk and reducing cone yield. (2) Methods: Five Beauveria bassiana strains were evaluated for virulence against fourth-instar larvae. And the levels of T-AOC and MDA in the larvae infected by each strain were measured. To assess larval responses to different strains, we measured the activities of six enzymes (SOD, CAT, POD, PPO, CarE, GST) and the levels of GSH and H₂O₂ in larvae treated with each strain. Additionally, the infection process of highly pathogenic B. bassiana in larvae was explored using scanning electron microscopy (SEM). (3) Results: Strain CGMCC3.2055 demonstrated the highest toxicity to larvae, achieving a cumulative corrected mortality of 80.56% on the 4th day and an LT₅₀ of 3.248 days. The T-AOC of larvae treated with strain CGMCC3.2055 was inhibited within 48 h. The relative MDA content in this group was significantly higher than that in other strain-treated groups at 6, 12, and 24 h. In Bb01-treated larvae, H₂O₂ accumulation at 6 and 24 h post-infection was influenced by POD activity rather than GSH levels; in BbZ1-treated larvae, the activities of CAT and POD were upregulated at 6 and 36 h, while the activity of SOD was downregulated, but the content of H₂O₂ increased significantly, resulting in accumulation; in CFCC81428-treated larvae, a decline in T-AOC coincided with substantial H₂O₂ accumulation over 48 h, while a concomitant increase in GSH content bolstered tolerance to lethal oxidative damage; in CGMCC3.2055-treated larvae, H₂O₂ only accumulated significantly at 24 and 48 h, yet upregulated CAT and POD were insufficient to effectively scavenge the excess H₂O₂; and in bio-21738-treated larvae, SOD-driven dismutation generated substantial H₂O₂ from 12 to 48 h, leading to pronounced accumulation from 6 to 48 h, yet limited upregulation of POD (only at 6 and 12 h) and CAT (only at 12 and 48 h) were insufficient to mitigate H₂O₂ buildup. PPO activity was upregulated within 48 h in all treatment groups except for BbZ1, where no upregulation was observed at 12 and 48 h. GST activity was upregulated in all treatment groups except for CGMCC3.2055, where a downregulation was observed at 12 h post-infection. CarE activity was significantly upregulated within 48 h in both CFCC81428 and CGMCC3.2055 groups; in the Bb01 group, CarE was upregulated only at 6 and 48 h; in the BbZ1 group, CarE was downregulated only at 48 h; and in the bio-21738 group, CarE showed no upregulation at 24 and 48 h. Through SEM, the infection process of the strain CGMCC3.2055 on the surface of the larvae was further determined, which mainly included adhesion, the appearance of bud-like protrusions, the growth of germ tubes along the epidermis and penetration of the epidermis, as well as the colonization of the strain and its emergence from the surface of the larvae. (4) Conclusions: This study first screened the highly pathogenic B. bassiana strain CGMCC3.2055 by evaluating its virulence to larvae and post-infection T-AOC and MDA levels. It also clarified the strain’s infection process and the larvae’s immune responses to various strains. Full article

In insect–microbe symbiosis, understanding the diversity of associated bacteria is crucial. DNA-dependent sequence methods are widely used to assess microbial diversity in insects, but they cannot distinguish between live and dead microbes. In contrast, RNA-dependent sequencing can identify alive bacterial communities, making them more suitable for evaluating alive microbiota diversity. However, its practical reliability in insect–microbe symbiosis remains poorly validated. This study investigated larval gut bacteria diversity of Delia antiqua, a major pest of Liliaceae crops, by employing both DNA- and RNA-dependent 16S rRNA amplicon sequencing. The reliability of both sequencing methods was evaluated by comparing the effects of synthetic communities (SynComs, constructed according to DNA- or RNA-dependent sequencing) and bacterial communities from wild larvae on axenic larvae. Results revealed significant differences in bacterial community between DNA- and RNA-dependent sequence samples. Compared to bacterial communities from wild larvae, the SynCom constructed based on RNA-dependent sequencing exhibited inhibition effects on D. antiqua larvae survival and body weight, while DNA-dependent SynCom did not, suggesting that DNA-dependent methods were superior for assessing symbiotic microbiota in D. antiqua. This work will provide insights into microbial diversity detection in D. antiqua and offer a framework for other insect–microbe studies. Full article

Domestication significantly altered the phenotypic and chemical traits of murtilla, notably reducing the emission of volatile compounds essential for plant–insect interactions. This reduction may affect the plant’s natural defense mechanisms, influencing its interactions with herbivores and predators. Therefore, this study tests whether domestication reduces volatile emissions in murtilla, increasing aphid preference and decreasing lacewing attraction. We selected wild ancestors (19-1, 22-1, and 23-2) from a longterm Ugni molinae germplasm bank. Crosses between these wild ancestors generated four first-generation domesticated ecotypes, 10-1, 16-16, 17-4, and 66-2, used in this study. These first-generation domesticated ecotypes were six years old at the time of the study and were used for comparisons in volatile profile and insect interaction analyses. The olfactometric preference index (OPI) for lacewing larvae and aphids revealed that wild ancestors attracted more predators than domesticated plants. For example, Ecotype 19-1 had an OPI of 1.64 for larvae and 1.49 for aphids, while Ecotype 10-1 showed lower attraction (OPI of 1.01 for larvae and 1.00 for aphids). Gas chromatography analysis identified differences in volatile organic compounds, with wild ancestor ecotypes emitting higher levels of compounds such as 2-hexanone, 1,8-cineole, and α-caryophyllene. Principal component analysis and hierarchical clustering confirmed these chemical distinctions. In olfactometer assays, lacewing larvae preferred α-caryophyllene and 2,4-dimethyl acetophenone, while aphids favored 2-hexanone and 3-hexanol. In Y-tube assays, lacewing adults showed strong attraction to α-pinene and 2,4-dimethyl acetophenone, with preferences increasing with concentration. These results indicate that domestication altered the volatile murtilla profile, reducing its attractiveness to natural predators while increasing its susceptibility to herbivores, supporting the plant domestication defense theory. Full article

Spodoptera frugiperda, a major global agricultural pest, poses significant challenges to chemical control methods due to pesticide resistance and environmental concerns, underscoring the need for sustainable management strategies. Attractants based on host plant volatiles offer a promising eco-friendly approach, but their development for S. frugiperda is hindered by limited research on host recognition mechanisms. This study reveals that female S. frugiperda preferentially oviposit on maize at the seedling stage. Using electrophysiological techniques, we identified p-xylene and (+)-camphor from seedling-stage maize volatiles as key compounds eliciting strong responses in female S. frugiperda. Behavioral assays confirmed that these compounds (p-xylene at the concentration of 5%, 10%, and 20% and (+)-camphor at 1%, 5%, and 10%) significantly attract females, establishing them as the key odor cues for host selection. Moreover, these volatiles are more abundant in seedling-stage maize, suggesting that S. frugiperda assesses maize growth stages based on their concentrations. Importantly, larvae reared on seedling-stage maize exhibited higher survival rates than those on later-stage maize, indicating that oviposition site selection directly affects offspring fitness. These findings demonstrate that S. frugiperda uses p-xylene and (+)-camphor to evaluate maize development and select suitable oviposition sites, thereby enhancing larval survival. This study provides a foundation for developing targeted attractants for S. frugiperda and highlights the seedling stage as a critical period for implementing pest control measures, particularly in autumn maize production, given the higher pest population density during this phase. Full article