Stinging Ant Anaphylaxis: Advances in Diagnosis and Treatment.

Stinging ants represent a wide range of over 200 different species across the world, of which Solenopsis, Myrmecia, Pogonomyrmex, and Brachyponera genera account for a substantial economic and healthcare burden. S. invicta (red imported fire ant [IFA]) and M. pilosula (jack jumper ant [JJA]) are 2 species of high clinical importance, known to cause anaphylaxis in humans, with numerous reported fatalities. Diagnostic testing should be performed in patients with a history of a systemic reaction with skin testing and/or in vitro specific immunoglobulin E (IgE) testing. In vitro testing is commercially available for IFA through whole-body extract specific IgE and JJA venom-specific IgE, but not widely available for other stinging ant species. Commercial venom component testing for IFA and JJA is currently not available. Patients with a clinical history and positive specific IgE testing should undergo treatment with specific immunotherapy, which is currently available for IFA and JJA. Buildup may be performed using conventional, semi-rush, rush, or ultra-rush schedules with similar risk profiles for IFA. Optimal duration for whole=body extract immunotherapy for IFA and specific JJA venom immunotherapy is not well studied, but generally recommended for at least 3 to 5 years. Sting challenges are used in research settings, primarily to assess treatment efficacy of immunotherapy.

Effect of entomopathogenic fungi on behavior and physiology of Solenopsis invicta (Hymenoptera, Formicidae).

In an ant colony, a large number of nestmates with a similar gene pool coexist, making them more vulnerable to pathogenic attacks. These pathogens influence the behavior and physiology of the fire ant Solenopsis invicta Buren. Here, we evaluated the impact of entomopathogenic fungi (EPF) Metarhizium anisopliae on the behavior (locomotion and foraging) and physiology (biological molecules, anti-fungal activity, and survival) of S. invicta. Distance traveled and velocity significantly decreased, while turn angle and angular velocity significantly increased in ants exposed to a higher concentration of M. anisopliae compared to ants exposed to control after 36 h, which showed disturbed locomotion. Fungus infection significantly affected the foraging behavior of ants. Fungus-exposed ants spent significantly less time in the food zone (area with food) than in the inner zone (area without food). The activities of 4 enzymes, peroxidase, glutathione-S-transferase, hydrogen peroxide (H2O2), and carboxylesterase were significantly decreased. In contrast, catalase and anti-fungal activities were increased after fungal exposure compared to the control. The activity of acetylcholinesterase, which hydrolyses the important neurotransmitter acetylcholine, also decreased after fungal application compared to the control. Survival of ants was also significantly reduced after fungus infection compared to the control. Our findings help to understand the influence of M. anisopliae on the behavior and physiology of S. invicta, which will help in the management of S. invicta using the EPF M. anisopliae.

The Human Health Impacts of the Red Imported Fire Ant in the Western Pacific Region Context: A Narrative Review.

Background: The red imported fire ant (RIFA) is one of the world's most destructive invasive species. RIFA stings are painful and can lead to allergic reactions, including life-threatening anaphylaxis, yet health impacts remain inadequately defined. Methods: We searched MEDLINE (Ovid) and Google Scholar (grey literature) from inception until 20 September 2023 for articles in English using search terms related to red imported fire ants and allergies, including anaphylaxis. Results: Approximately a third of the population in RIFA-infested areas are stung each year. The most frequent reaction is a sterile 1-2 mm pseudo pustule on the skin. Approximately 20% of stings cause a large local reaction and between about 0.5% and 2% stings cause a systemic allergic reaction which can range from skin symptoms to life-threatening anaphylaxis. Local biodiversity is also significantly disrupted by invading RIFA and may lead to complex adverse effects on human health, from agriculture losses to expanded ranges for pathogen vectors. Conclusions: The potential for red imported fire ants to establish themselves as an invasive species in the Western Pacific presents a substantial and costly health issue. Successful eradication and surveillance programs, to identify and eradicate new incursions, would avoid substantial health impacts and costs.

Misapplied management makes matters worse: Spatially explicit control leverages biotic interactions to slow invasion.

A wide range of approaches has been used to manage the spread of invasive species, yet invaders continue to be a challenge to control. In some cases, management actions have no effect or may even inadvertently benefit the targeted invader. Here, we use the mid-20th century management of the Red Imported Fire Ant, Solenopsis invicta, in the US as a motivating case study to explore the conditions under which such wasted management effort may occur. Introduced in approximately 1940, the fire ant spread widely through the southeast US and became a problematic pest. Historically, fire ants were managed with broad-spectrum pesticides; unfortunately, these efforts were largely unsuccessful. One hypothesis suggests that, by also killing native ants, mass pesticide application reduced competitive burdens thereby enabling fire ants to invade more quickly than they would in the absence of management. We use a mechanistic competition model to demonstrate the landscape-level effects of such management. We explicitly model the extent and location of pesticide applications, showing that the same pesticide application can have a positive, neutral, or negative effect on the progress of an invasion, depending on where it is applied on the landscape with respect to the invasion front. When designing management, the target species is often considered alone; however, this work suggests that leveraging existing biotic interactions, specifically competition with native species, can increase the efficacy of management. Our model not only highlights the potential unintended consequences of ignoring biotic interactions, but also provides a framework for developing spatially explicit management strategies that take advantage of these biotic interactions to work smarter, not harder.

N6-methyladenosine modification of RNA controls dopamine synthesis to influence labour division in ants.

The N6-methyladenosine (m6A) modification of RNA has been reported to remodel gene expression in response to environmental conditions; however, the biological role of m6A in social insects remains largely unknown. In this study, we explored the role of m6A in the division of labour by worker ants (Solenopsis invicta). We first determined the presence of m6A in RNAs from the brains of worker ants and found that m6A methylation dynamics differed between foragers and nurses. Depletion of m6A methyltransferase or chemical suppression of m6A methylation in foragers resulted in a shift to 'nurse-like' behaviours. Specifically, mRNAs of dopamine receptor 1 (Dop1) and dopamine transporter (DAT) were modified by m6A, and their expression increased dopamine levels to promote the behavioural transition from foragers to nurses. The abundance of Dop1 and DAT mRNAs and their stability were reduced by the inhibition of m6A modification caused by the silencing of Mettl3, suggesting that m6A modification in worker ants modulates dopamine synthesis, which regulates labour division. Collectively, our results provide the first example of the epitranscriptomic regulation of labour division in social insects and implicate m6A regulatory mechanism as a potential novel target for controlling red imported fire ants.

2-Methoxybenzaldehyde effectively repels ants.

Ants can particularly make for harmful pests, infesting human homes and reducing crop yields. The damage caused by ants and the efforts to mitigate the damage are hugely costly. Broad-spectrum insecticides are used most commonly; however, due to their negative side effects, there is increasing interest in nontoxic alternatives. One promising commercially available alternative is 2-hydroxybenzaldehyde, which is naturally produced by various arthropods as a means of chemical defense and effectively repels ants. Here we conduct a structure-activity relationship investigation, testing how different chemical modifications alter the repellence of 2-hydroxybenzaldehyde. We find that 2-methoxybenzaldehyde is considerably more effective than 2-hydroxybenzaldehyde at repelling the common black garden ant, Lasius niger. We next compare the most effective repellent chemicals against 4 particularly harmful ant species to confirm that the results obtained with L. niger are general to ants and that our results are relevant to mitigate the costs of ant damage.

The temporal-spatial expression and functional analysis of three gustatory receptor genes in Solenopsis invicta using sweet and bitter compounds.

The insect gustatory system participates in identifying potential food sources and avoiding toxic compounds. During this process, gustatory receptors (GRs) recognize feeding stimulant and deterrent compounds. However, the GRs involved in recognizing stimulant and deterrent compounds in the red imported fire ant, Solenopsis invicta, remain unknown. Therefore, we conducted a study on the genes SinvGR1, SinvGR32b, and SinvGR28a to investigate the roles of GRs in detecting feeding stimulant and deterrent compounds. In this current study, we found that sucrose and fructose are feeding stimulants and the bitter compound quinine is a feeding deterrent. The fire ant workers showed significant behavior changes to avoid the bitter taste in feeding stimulant compounds. Reverse transcription quantitative real-time polymerase chain reaction results from developmental stages showed that the SinvGR1, SinvGR32b, and SinvGR28a genes were highly expressed in fire ant workers. Tissue-specific expression profiles indicated that SinvGR1, SinvGR32b, and SinvGR28a were specifically expressed in the antennae and foreleg tarsi of workers, whereas SinvGR32b gene transcripts were also highly accumulated in the male antennae. Furthermore, the silencing of SinvGR1 or SinvGR32b alone and the co-silencing of both genes disrupted worker stimulation and feeding on sucrose and fructose. The results also showed that SinvGR28a is required for avoiding quinine, as workers with knockdown of the SinvGR28a gene failed to avoid and fed on quinine. This study first identified stimulant and deterrent compounds of fire ant workers and then the GRs involved in the taste recognition of these compounds. This study could provide potential target gustatory genes for the control of the fire ant.

A Pyr-loaded polymer microparticle for effectively controlling Solenopsis invicta (Hymenoptera: Formicidae) in the nest.

Human interference and incorrect use of pesticides are easy to induce red imported fire ant (RIFA) escape and migrate from a nest, resulting in ineffective control of RIFA. In order to avoid RIFA alert, we designed an amphiphilic PSI-mPEG-Boc-DAH loaded Pyr to make the microparticles with effective controlled release. The investigation showed that the quantity of Pyr released by Pyr@PSI-mPEG-Boc-DAH under acidic environment was only 36.40 ± 1.90% at 48 h, whereas the release rate of original Pyr was 75.23 ± 5.71%. And the RIFA mortality rate of 1 ppm Pyr in Pyr@PSI-mPEG-Boc-DAH microparticles at 48 h was only 7.78%, which was significantly lower than that of the Pyr (47.78%). Futhermore, the death rate increased sharply after 48 h, and reached 95.84% within a week after using Pyr@PSI-mPEG-Boc-DAH microparticles. Moreover, PSI-mPEG-Boc-DAH carriers could be absorbed and even transported to crop of the RIFA for subsequent trophallaxis by using fluorescence tracking. In the field experiment, the reduction rate of Pyr@PSI-mPEG-Boc-DAH treatment was achieved 99.89% after 7 d. Pyr@PSI-mPEG-Boc-DAH didn't cause RIFA to be alarmed within 48 h and could kill nearly all of ants in the nest after 7 d, which showed a very good control effect in the field experiment. This work provided a new idea and guidance for the effective control RIFA and the development of sustainable agriculture.

Enhancement of tolerance against flonicamid in Solenopsis invicta (Hymenoptera: Formicidae) through overexpression of CYP6A14.

Solenopsis invicta is a main issue in southern China and is causing significant damage to the local ecological environment. The extensive use of insecticides has resulted in the development of tolerance in S. invicta. In our study, ten S. invicta colonies from Sichuan Province exhibited varying degrees of tolerance against flonicamid, with LC50 values from 0.49 mg/L to 8.54 mg/L. The sensitivity of S. invicta to flonicamid significantly increased after treatment with the P450 enzyme inhibitor piperonyl butoxide (PBO). Additionally, the activity of P450 in S. invicta was significantly enhanced after being treated with flonicamid. Flonicamid induced the expression levels of CYP4aa1, CYP9e2, CYP4C1, and CYP6A14. The expression levels of these P450 genes were significantly higher in the tolerant colonies compared to the sensitive colonies, and the relative copy numbers of CYP6A14 in the tolerant colonies were 2.01-2.15 fold. RNAi feeding treatment effectively inhibited the expression of P450 genes, thereby reducing the tolerance of S. invicta against flonicamid. In addition, the overexpression of CYP6A14 in D. melanogaster resulted in reduced sensitivity to flonicamid. Our investigations revealed hydrophobic interactions between flonicamid and seven amino acid residues of CYP6A14, along with the formation of a hydrogen bond between Glu306 and flonicamid. Our findings suggest that flonicamid can effectively control S. invicta and P450 plays a pivotal role in the tolerance of S. invicta against flonicamid. The overexpression of CYP6A14 also increased tolerance to flonicamid.

Comparative Analyses of Reproductive Caste Types Reveal Vitellogenin Genes Involved in Queen Fertility in Solenopsis invicta.

The red imported fire ant (Solenopsis invicta Buren) is a social pest species with a robust reproductive ability that causes extensive damage. Identification of the genes involved in queen fertility is critical in order to better understand the reproductive biology and screening for the potential molecular targets in S. invicta. Here, we used the mRNA deep sequencing (RNA-seq) approach to identify differentially expressed genes (DEGs) in the transcriptomes of three reproductive caste types of S. invicta, including queen (QA) and winged female (FA) and male (MA) ants. The genes that were specific to and highly expressed in the queens were then screened, and the Vg2 and Vg3 genes were chosen as targets to explore their functions in oogenesis and fertility. A minimum of 6.08 giga bases (Gb) of clean reads was obtained from all samples, with a mapping rate > 89.78%. There were 7524, 7133, and 977 DEGs identified in the MA vs. QA, MA vs. FA, and FA vs. QA comparisons, respectively. qRT-PCR was used to validate 10 randomly selected DEGs, including vitellogenin 2 (Vg2) and 3 (Vg3), and their expression patterns were mostly consistent with the RNA-seq data. The S. invicta Vgs included conserved domains and motifs that are commonly found in most insect Vgs. SiVg2 and SiVg3 were highly expressed in queens and winged females and were most highly expressed in the thorax, followed by the fat body, head, and epidermis. Evaluation based on a loss-of-function-based knockdown analysis showed that the downregulation of either or both of these genes resulted in smaller ovaries, less oogenesis, and less egg production. The results of transcriptional sequencing provide a foundation for clarifying the regulators of queen fertility in S. invicta. The functions of SiVg2 and SiVg3 as regulators of oogenesis highlight their importance in queen fecundity and their potential as targets of reproductive disruption in S. invicta control.

Effect of Juvenile Hormone on Worker Behavioral Transition in the Red Imported Fire Ant, Solenopsis invicta (Hymenoptera: Formicidae).

The division of labor among workers is a defining characteristic of social insects and plays a pivotal role in enhancing the competitive advantage of their colony. Juvenile hormone (JH) has long been hypothesized to be the essential driver in regulating the division of labor due to its ability to accelerate behavioral transitions in social insects, such as honeybees. The regulation of behavioral transitions by JH in the red imported fire ant (RIFA), Solenopsis invicta, a typical social pest, is unclear. Through video capture and analysis, we investigated the effects of the juvenile hormone analogue (JHA) methoprene on brood care, phototaxis behavior, and threat responsiveness of RIFA nurse workers. Our results showed that the JHA application significantly reduced the time and frequency of brood care behavior by nurse workers while increasing their walking distance and activity time in the light area. Additionally, the application of JHA made ants become excited, indicating a significant improvement in their activity level (movement distance, time, and speed). Furthermore, it was observed that the application of JHA did not affect the threat responsiveness of nurse workers towards stimuli (nestmates or non-nestmates). Our study demonstrates that the application of JHA reduced brood care behavior and enhanced phototaxis in nurse workers, which may reveal the role of JH in facilitating behavioral transitions in RIFA from intranidal tasks to extranidal activity. This study provides an experimental basis for further elucidating the mechanism underlying the division of labor in social insects.

Foraging Behaviors of Red Imported Fire Ants (Hymenoptera Formicidae) in Response to Bait Containing Different Concentrations of Fipronil, Abamectin, or Indoxacarb.

The red imported fire ant, Solenopsis invicta Buren, is a severe pest with agricultural, ecological, and medical significance. The baiting treatment is one of the main methods to control S. invicta. However, few studies have evaluated the acceptance of fire ant bait. Here, field and laboratory studies were conducted to investigate the foraging behaviors of S. invicta responding to fire ant baits containing different concentrations of active ingredients (fipronil, abamectin, or indoxacarb). Field studies showed that S. invicta transported significantly less 0.0125% fipronil bait than control bait (without toxicant) and 0.0001% fipronil bait. The number of foraging ants significantly decreased with an increase in fipronil concentration. Our previous study showed that S. invicta usually buries the food treated with repellent chemicals, and interestingly, significantly more soil particles were transported into tubes containing 0.0001% fipronil bait than tubes containing control bait or 0.0125% fipronil bait. In addition, S. invicta transported significantly less 0.0005% abamectin bait than control bait, and significantly fewer ants were found in tubes containing 0.0125% abamectin bait than control bait. However, there was no significant difference in bait transport, number of foraging ants, and weight of soil particles relocated in tubes containing different concentrations of indoxacarb bait. In addition, laboratory studies showed that S. invicta transported significantly less 0.0125% fipronil bait than control bait and bait containing abamectin (0.0025% or 0.0125%) or indoxacarb (0.0125% or 0.0625%). In addition, the transport speed for the 0.0125% fipronil bait was the slowest. These results show that specific concentrations of some active ingredients may negatively affect bait acceptance for S. invicta, and should be avoided in fire ant bait production.

Baccharis Species Essential Oils: Repellency and Toxicity against Yellow Fever Mosquitoes and Imported Fire Ants.

Essential oils from five Baccharis species were screened for their toxicity and biting deterrence/repellency against yellow fever mosquito, Aedes aegypti (L.), and imported fire ants, including Solenopsis invicta Buren (RIFA), Solenopsis richteri Forel (BIFA) and their hybrids (HIFA). Baccharis microdonta DC. and B. punctulata DC. at 10 µg/cm2 showed biting deterrence similar to DEET, N, N-diethyl-meta-toluamide at 25 nmol/cm2, whereas the repellency of B. pauciflosculosa DC., B. sphenophylla Dusén ex Malme and B. reticularioides Deble & A.S. Oliveira essential oils was significantly lower than DEET against mosquitoes. Two major compounds from the active essential oils, kongol and spathulenol, also exhibited biting deterrence similar to DEET against mosquitoes. The highest toxicity exhibited against mosquitoes was by Baccharis punctulata essential oil (LC50 = 20.4 ppm), followed by B. pauciflosculosa (LC50 = 31.9 ppm), B. sphenophylla (LC50 = 30.8 ppm), B. microdonta (LC50 = 28.6 ppm), kongol (LC50 = 32.3 ppm), spathulenol (LC50 = 48.7 ppm) and B. reticularioides essential oil (LC50 = 84.4 ppm). Baccharis microdonta essential oil showed repellency against RIFA, BIFA and HIFA at 4.9, 4.9 and 39 µg/g, respectively. Baccharis microdonta essential oil also showed toxicity with LC50 of 78.9, 97.5 and 136.5 µg/g against RIFA, BIFA and HIFA, respectively, at 24 h post treatment.

Co-Occurrence of Wing Deformity and Impaired Mobility of Alates with Deformed Wing Virus in Solenopsis invicta Buren (Hymenoptera: Formicidae).

Deformed wing virus (DWV), a major honey bee pathogen, is a generalist insect virus detected in diverse insect phyla, including numerous ant genera. Its clinical symptoms have only been reported in honey bees, bumble bees, and wasps. DWV is a quasispecies virus with three main variants, which, in association with the ectoparasitic mite, Varroa destructor, causes wing deformity, shortened abdomens, neurological impairments, and colony mortality in honey bees. The red imported fire ant, Solenopsis invicta Buren, is one of the most-invasive and detrimental pests in the world. In this study, we report the co-occurrence of DWV-like symptoms in S. invicta and DWV for the first time and provide molecular evidence of viral replication in S. invicta. Some alates in 17 of 23 (74%) lab colonies and 9 of 14 (64%) field colonies displayed deformed wings (DWs), ranging from a single crumpled wing tip to twisted, shriveled wings. Numerous symptomatic alates also exhibited altered locomotion ranging from an altered gait to the inability to walk. Deformed wings may prevent S. invicta alates from reproducing since mating only occurs during a nuptial flight. The results from conventional RT-PCR and Sanger sequencing confirmed the presence of DWV-A, and viral replication of DWV was confirmed using a modified strand-specific RT-PCR. Our results suggest that S. invicta can potentially be an alternative and reservoir host for DWV. However, further research is needed to determine whether DWV is the infectious agent that causes the DW syndrome in S. invicta.

Repellency of Carvacrol, Thymol, and Their Acetates against Imported Fire Ants.

In the United States, imported fire ants are commonly referred to as red imported fire ants (Solenopsis invicta Buren), black imported fire ants (S. richteri Forel), and hybrid imported fire ants (S. invicta × S. richteri). They are significant pests, and their control heavily relies on synthetic insecticides. The extensive use of insecticides has led to public concern about their potential negative effects on human health and the well-being of wildlife and the environment. As an alternative, plant-derived natural compounds, particularly essential oils (EOs) and their main constituents, show promise as safe and environmentally friendly products for controlling fire ants. Repellants are useful in managing fire ants, and plant-derived natural repellants may serve as a safer and more environmentally friendly option. This study investigates the repellency of EO-derived compounds carvacrol, thymol, and their acetates against imported fire ant workers. The results revealed that carvacrol, a GRAS compound (Generally Recognized As Safe), was the most potent repellent against S. invicta, S. richteri, and their hybrid, with minimum repellent effective doses (MREDs) of 0.98 µg/g, 7.80 µg/g, and 0.98 µg/g, respectively. Thymol also exhibited strong repellency, with MREDs of 31.25 µg/g, 31.25 µg/g, and 7.8 µg/g, respectively. Furthermore, thyme-red essential oil, characterized by a thymol chemotype containing 48.8% thymol and 5.1% carvacrol, was found to effectively repel the hybrid ants with an MRED of 15.6 µg/g. In contrast, thyme essential oil, characterized by a linalool chemotype lacking thymol and carvacrol, did not exhibit any repellent effect, even at the highest tested dose of 125 µg/g. This study provides the first evidence of the potent repellency of carvacrol and thymol against imported fire ant workers, indicating their potential as promising repellents for fire ant control.

Transcriptomic Analysis of Metarhizium anisopliae-Induced Immune-Related Long Non-Coding RNAs in Polymorphic Worker Castes of Solenopsis invicta.

Long non-coding RNAs (lncRNAs) represent a class of RNA molecules that do not encode proteins. Generally studied for their regulatory potential in model insects, relatively little is known about their immunoregulatory functions in different castes of eusocial insects, including Solenopsis invicta, a notoriously invasive insect pest. In the current study, we used Metarhizium anisopliae, an entomopathogenic fungus, to infect the polymorphic worker castes (Major and Minor Workers) and subjected them to RNA sequencing at different intervals (6, 24, and 48 h post-infection (hpi)). Comprehensive bioinformatic analysis identified 5719 (1869 known and 3850 novel) lncRNAs in all libraries. Genomic characteristics analysis showed that S. invicta lncRNAs exhibited structural similarities with lncRNAs from other eusocial insects, including lower exon numbers, shorter intron and exon lengths, and a lower expression profile. A comparison of lncRNAs in major and minor worker ants revealed that several lncRNAs were exclusively expressed in one worker caste and remained absent in the other. LncRNAs such as MSTRG.12029.1, XR_005575440.1 (6 h), MSTRG.16728.1, XR_005575440.1 (24 h), MSTRG.20263.41, and MSTRG.11994.5 (48 h) were only present in major worker ants, while lncRNAs such as MSTRG.8896.1, XR_005574239.1 (6 h), MSTRG.20289.8, XR_005575051.1 (24 h), MSTRG.20289.8, and MSTRG.6682.1 (48 h) were only detected in minor workers. Additionally, we performed real-time quantitative PCR and experimentally validated these findings. Functional annotation of cis-acting lncRNAs in major worker ants showed that lncRNAs targeted genes such as serine protease, trypsin, melanization protease-1, spaetzle-3, etc. In contrast, apoptosis and autophagy-related genes were identified as targets of lncRNAs in minor ants. Lastly, we identified several lncRNAs as precursors of microRNAs (miRNAs), such as miR-8, miR-14, miR-210, miR-6038, etc., indicating a regulatory relationship between lncRNAs, miRNAs, and mRNAs in antifungal immunity. These findings will serve as a genetic resource for lncRNAs in polymorphic eusocial ants and provide a theoretical basis for exploring the function of lncRNAs from a unique and novel perspective.

Transcriptomic Analysis Reveals the Impact of the Biopesticide Metarhizium anisopliae on the Immune System of Major Workers in Solenopsis invicta.

The red imported fire ant (Solenopsis invicta Buren, 1972) is a globally significant invasive species, causing extensive agricultural, human health, and biodiversity damage amounting to billions of dollars worldwide. The pathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (1883), widely distributed in natural environments, has been used to control S. invicta populations. However, the interaction between M. anisopliae and the immune system of the social insect S. invicta remains poorly understood. In this study, we employed RNA-seq to investigate the effects of M. anisopliae on the immune systems of S. invicta at different time points (0, 6, 24, and 48 h). A total of 1313 differentially expressed genes (DEGs) were identified and classified into 12 expression profiles using short time-series expression miner (STEM) for analysis. Weighted gene co-expression network analysis (WGCNA) was employed to partition all genes into 21 gene modules. Upon analyzing the statistically significant WGCNA model and conducting Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the modules, we identified key immune pathways, including the Toll and Imd signaling pathways, lysosomes, autophagy, and phagosomes, which may collectively contribute to S. invicta defense against M. anisopliae infection. Subsequently, we conducted a comprehensive scan of all differentially expressed genes and identified 33 immune-related genes, encompassing various aspects such as recognition, signal transduction, and effector gene expression. Furthermore, by integrating the significant gene modules derived from the WGCNA analysis, we constructed illustrative pathway diagrams depicting the Toll and Imd signaling pathways. Overall, our research findings demonstrated that M. anisopliae suppressed the immune response of S. invicta during the early stages while stimulating its immune response at later stages, making it a potential biopesticide for controlling S. invicta populations. These discoveries lay the foundation for further understanding the immune mechanisms of S. invicta and the molecular mechanisms underlying its response to M. anisopliae.

Regioselective Claisen-Schmidt Adduct of 2-Undecanone from Houttuynia cordata Thunb as Insecticide/Repellent against Solenopsis invicta and Repositioning Plant Fungicides against Colletotrichum fragariae.

The U.S. Department of Agriculture (USDA) has established research programs to fight the phytopathogen Colletotrichum fragariae and the invasive red imported fire ant, Solenopsis invicta. C. fragariae is known to cause anthracnose disease in fruits and vegetables, while S. invicta is known for its aggressive behavior and painful stings and for being the cause of significant damage to crops, as well as harm to humans and animals. Many plants have been studied for potential activity against C. fragariae and S. invicta. Among the studied plants, Houttuynia cordata Thunb has been shown to contain 2-undecanone, which h is known for its antifungal activity against Colletotrichum gloesporioides. Based on the mean amount of sand removed, 2-undecanone showed significant repellency at 62.5 µg/g, similar to DEET (N,N-diethyl-meta-toluamide), against S. invicta. The 2-Undecanone with an LC50 of 44.59 µg/g showed toxicity against S. invicta workers. However, neither H. cordata extract nor 2-undecanone had shown activity against C. fragariae despite their known activity against C. gloesporioides, which in turn motivates us in repositioning 2-undecanone as a selected candidate for a Claisen-Schmidt condensation that enables access to several analogs (2a-f). Among the prepared analogs, (E)-1-(3-methylbenzo[b]thiophen-2-yl)dodec-1-en-3-one (2b) and (E)-1-(5-bromothiophen-2-yl)dodec-1-en-3-one (2f) showed promising activity against C. fragariae, revealing a distinctive structural activity relationship (SAR). The generated analogs revealed a clear regioselectivity pattern through forming the C=C alkene bond at position C-1. These data open the window for further lead optimization and product development in the context of managing C. fragariae and S. invicta.

Labor division of worker ants can be controlled by insulin synthesis targeted through miR-279c-5p in Solenopsis invicta (Hymenoptera: Formicidae).

BACKGROUND: In social insects, the labor division of workers is ubiquitous and controlled by genetic and environmental factors. However, how they modulate this coordinately remains poorly understood. RESULTS: We report miR-279c-5p participation in insulin synthesis and behavioral transition by negatively regulating Rab8A in Solenopsis invicta. Eusocial specific miR-279c-5p is age-associated and highly expressed in nurse workers, and localized in the cytoplasm of neurons, where it is partly co-localized with its target, Rab8A. We determined that miR-279c-5p agomir suppressed Rab8A expression in forager workers, consequently decreasing insulin content, resulting in the behavioral shift to 'nurse-like' behaviors, while the decrease in miR-279c-5p increased Rab8A expression and increased insulin content in nurse workers, leading to the behavioral shift to 'foraging-like' behaviors. Moreover, insulin could rescue the 'foraging behavior' induced by feeding miR-279c-5p to nurse workers. The overexpression and suppression of miR-279c-5p in vivo caused an obvious behavioral transition between foragers and nurses, and insulin synthesis was affected by miR-279c-5p by regulating the direct target Rab8A. CONCLUSION: We first report that miR-279c-5p is a novel regulator that promotes labor division by negatively regulating the target gene Rab8A by controlling insulin production in ants. This miRNA-mediated mechanism is significant for understanding the behavioral plasticity of social insects between complex factors and potentially provides new targets for controlling red imported fire ants. © 2023 Society of Chemical Industry.

Biological Activities and Ecological Significance of Fire Ant Venom Alkaloids.

Venoms produced by arthropods act as chemical weapons to paralyze prey or deter competitors. The utilization of venom is an essential feature in the biology and ecology of venomous arthropods. Solenopsis fire ants (Hymenoptera: Formicidae) are medically important venomous ants. They have acquired different patterns of venom use to maximize their competitive advantages rendered by the venom when facing different challenges. The major components of fire ant venom are piperidine alkaloids, which have strong insecticidal and antibiotic activities. The alkaloids protect fire ants from pathogens over the course of their lives and can be used to defend them from predators and competitors. They are also utilized by some of the fire ants' natural enemies, such as phorid flies to locate host ants. Collectively, these ants' diverse alkaloid compositions and functions have ecological significance for their survival, successful invasion, and rapid range expansion. The venom alkaloids with powerful biological activities may have played an important role in shaping the assembly of communities in both native and introduced ranges.