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Aim Pathogens can play an important role in biological invasions. Introduced populations may be particularly vulnerable to pathogens due to factors such as low genetic diversity and high population density. However, introduced populations that escape their natural pathogens may reallocate resources away from immunity and towards growth and reproduction. Interestingly, introduced ants have been suggested to have increased tolerance to new pathogens, contributing to their success as introduced species. In this study, we aimed to investigate whether introduced Argentine ant populations harbour different viral loads compared to native populations and if these differences were related to immunity‐related gene expression. Location The study was conducted across the native range of Argentine ants in Argentina and four introduced regions in California, France, Australia and New Zealand. Methods We used RT‐qPCR to quantify viral loads and gene expression in the ants. We analysed 15 different potentially pathogenic viruses across the Argentine ant's native and introduced ranges. Results We found that five viruses, LhuPcV1, LhuPiLV1, LhuCV1, Kashmir Bee virus and LHUV‐1, presented high loads in Argentine ants compared to the other viruses we screened. We found a significant effect of range on viral infections: high viral loads were commonly found in ants from introduced populations, which also exhibited increased immune gene expression. We found highly significant correlations between viral loads and expression of immune and metabolic genes. However, these associations were not fully consistent across the studied regions, indicating the complexity of eco‐immunological phenomena. Main Conclusions Our results suggest that introduced Argentine ant populations host different viral communities compared to native populations and that these differences are correlated with changes in immunity‐related gene expression. The study highlights the complex role of pathogens in biological invasions and the importance of considering eco‐immunological factors when assessing the impact of introduced species.

The purpose of this study was to involve pest management professionals in the design of application techniques and strategies that would be efficacious and also reduce insecticide runoff. Our study involved measuring both the efficacy of treatments for the Argentine ant, Linepithema humile (Mayr), and the concurrent runoff of fipronil and pyrethroids. Two collaborating companies used low-impact protocols for controlling ants while minimizing runoff. Protocol 1 involved bimonthly treatments, while Protocol 2 was monthly. Both protocols involved an initial treatment with a fipronil spray around the foundation. At the garage door-driveway interface, the fipronil application was done as a pin stream for Protocol 1, and as a crack and crevice application in the expansion joint near the garage for Protocol 2. Protocol 1 replaced most pyrethroid sprays with bifenthrin granules placed around bushes and away from the driveway. For the next treatment on day 63, Protocol 1 also included cyfluthrin spray treatments around the house foundation and crack and crevice applications around the edge of the driveway. For the first treatment in Protocol 2, the fipronil spray was supplemented with spot treatments of cyfluthrin. For subsequent Protocol 2 treatments, botanical insecticides were applied. For weeks 1 and 2 posttreatment combined, Protocol 1 had significantly higher reductions in ant numbers compared with Protocol 2. Thereafter there were no significant differences between the protocols. Runoff of bifenthrin from the granules used with Protocol 1 was much lower than in previous trials involving bifenthrin sprays. Day 1 fipronil runoff for Protocol 2 was significantly lower than that for Protocol 1. This difference may be because of the crack and crevice application applied in Protocol 2. Cyfluthrin runoff was minimal for Protocol 2, which involved spot treatments to supplement the fipronil on day 1, or the botanical insecticides for subsequent treatments. Protocol 1 had a large peak of cyfluthrin runoff at day 63 corresponding to their house and driveway treatments.

Ant management in urban and natural areas often relies on toxic baits. Liquid baits are highly attractive to pest ants because they mimic natural food sources such as honeydew and nectar, the principal dietary components of many ants. However, liquid bait use has been limited owing to the lack of bait dispensers that are effective, inexpensive, and easy to service. The current study evaluated the potential of water-storing crystals (Polyacrylamide spheres) to effectively deliver liquid thiamethoxam baits to laboratory colonies of Argentine ants, Linepithema humile Mayr. Results of laboratory trials show that bait crystals saturated in 25% sucrose solution containing 0.007% thiamethoxam are highly attractive to Argentine ants and highly effective against all castes and life stages, including workers, queens, and brood. Fresh bait crystals were highly effective and required approximately 2 d to kill all workers and approximately 6 d to achieve complete mortality in queens and brood. Results of bait aging tests show that the crystals lose approximately 70% of moisture in 8 h and the duration of outdoor exposure has a significant effect on moisture loss and subsequently bait acceptance and bait efficacy. A gradual decrease in mortality was observed for all castes and life stages as bait age increased. In general, fresh baits and those aged for <8 h retained their efficacy and caused substantial mortality. Baits aged longer than 8 h were substantially less attractive and less effective. Horizontal transfer tests examined the transfer of thiamethoxam from live treated donors to live untreated recipients. The results show that donor ants that obtain thiamethoxam by feeding on bait crystals effectively transfer it to untreated recipient ants. The level of secondary mortality depended on the donor:recipient ratio, with approximately 40% recipient worker mortality with the 1:5 ratio and 15% recipient worker mortality with 1:10 or 1:20 ratios. However, no queens died in any transfer tests, suggesting that multiple feedings from multiple donors may be necessary to produce queen mortality. The results of the transfer tests demonstrate the role of trophallaxis in the distribution of thiamethoxam and confirm that thiamethoxam is effectively transferred in Argentine ant colonies. The distribution of thiamethoxam within Argentine ant colonies was further examined using protein marking coupled with an enzyme-linked immunosorbent assay to detect the marker. The distribution of thiamethoxam was highly efficient, with 79 plus or minus 13% of workers testing positive at 15 min and 100 plus or minus 0% of workers testing positive at 6 h. In summary, the results of this study demonstrate that water-storing crystals effectively deliver thiamethoxam to all castes and life stages of Argentine ants and may offer an effective tool for Argentine ant management.

Outdoor residual sprays are among the most common methods for targeting pestiferous ants in urban pest management programs. If impervious surfaces such as concrete are treated with these insecticides, the active ingredients can be washed from the surface by rain or irrigation. As a result, residual sprays with fipronil and pyrethroids are found in urban waterways and aquatic sediments. Given the amount of insecticides applied to urban settings for ant control and their possible impact on urban waterways, the development of alternative strategies is critical to decrease the overall amounts of insecticides applied, while still achieving effective control of target ant species. Herein we report a \"pheromone-assisted technique\" as an economically viable approach to maximize the efficacy of conventional sprays targeting the Argentine ant. By applying insecticide sprays supplemented with an attractive pheromone compound, (Z)-9-hexadecenal, Argentine ants were diverted from nearby trails and nest entrances and subsequently exposed to insecticide residues. Laboratory experiments with fipronil and bifenthrin sprays indicated that the overall kill of the insecticides on Argentine ant colonies was significantly improved (57-142% increase) by incorporating (Z)-9-hexadecenal in the insecticide sprays. This technique, once it is successfully implemented in practical pest management programs, has the potential of providing maximum control efficacy with reduced amount of insecticides applied in the environment.

Pest control methods that can target pest species with limited environmental impacts are a conservation and economic priority. Species-specific pest control using RNA interference is a challenging but promising avenue in developing the next generation of pest management. We investigate the feasibility of manipulating a biological invader’s immune system using double-stranded RNA (dsRNA) in order to increase susceptibility to naturally occurring pathogens. We used the invasive Argentine ant as a model, targeting the immunity-associated genes Spaetzle and Dicer-1 with dsRNA. We show that feeding with Spaetzle dsRNA can result in partial target gene silencing for up to 28 days in the laboratory and 5 days in the field. Dicer-1 dsRNA only resulted in partial gene knockdown after 2 days in the laboratory. Double-stranded RNA treatments were associated with significant gene expression disruptions across immune pathways in the laboratory and to a lower extent in the field. In total, 12 viruses and four bacteria were found in these ant populations. Some changes in viral loads in dsRNA-treated groups were observed. For example, Linepithema humile Polycipivirus 2 (LhuPCV2) loads increased after 2 days of treatment with Spaetzle and Dicer-1 dsRNA treatments in the laboratory. After treatment with the dsRNA in the field, after 5 days the virus Linepithema humile toti-like virus 1 (LhuTLV1) was significantly more abundant. However, immune pathway disruption did not result in a consistent increase in microbial infections, nor did it alter ant abundance in the field. Some viruses even declined in abundance after dsRNA treatment. Our study explored the feasibility of lowering a pest’s immunity as a control tool. We demonstrate that it is possible to alter immune gene expression of pest species and pathogen loads, although in our specific system the affected pathogens did not appear to influence pest abundance. We provide suggestions on future directions for dsRNA-mediated immune disruption in pest species, including potential avenues to improve dsRNA delivery as well as the importance of pest and pathogen biology. Double-stranded RNA targeting immune function might be especially useful for pest control in systems in which viruses or other microorganisms are prevalent and have the potential to be pathogenic.

Aim Thousands of non‐native species have established populations and spread in outdoor environments (i.e., Naturalised), yet some populations or species only occur indoors, potentially due to unsuitable climates. We assessed the hypothesis that non‐native ants are more often restricted to indoor environments when they invade regions with climates dissimilar from their native regions. Furthermore, we forecasted how climate change could influence the naturalisation of indoor‐restricted non‐native ants. Location Global. Methods Using a global database of 323 non‐native ant taxa across 477 regions, we modelled how average climatic conditions in the native and invaded regions of each taxon determined whether they naturalised or were restricted indoors. We then modelled regional climatic suitability for the naturalisation of indoor‐restricted non‐native ants and projected future changes under climate change scenarios. We further assessed if climate change would facilitate the naturalisation of impactful non‐native ants using a global database describing their known impacts. Results Non‐native ants originating from warm regions were more likely restricted indoors when introduced to cold regions. Under 2°C and 4°C of warming, the number of indoor‐restricted non‐native ant species projected to find suitable regional climates for naturalisation increased by an average of 0.08 (maximum = 1.2) and 0.27 (maximum = 3.7) taxa per region, respectively. These anticipated naturalisations include high‐impact non‐native ants, such as the Argentine ant Linepithema humile and are expected to increase socioeconomic and environmental impacts under both warming scenarios, particularly in European regions. Main Conclusions Our findings suggest that indoor environments serve as microclimatic beachheads for biological invasions, especially in cold regions in the Northern Hemisphere. Failure to limit climate warming and inadequate biosecurity management in indoor environments may facilitate the naturalisation of non‐native ants, with costly repercussions on nature and society.

Biological invasions are a leading cause of global change, yet their long-term effects remain hard to predict. Invasive species can remain abundant for long periods of time, or exhibit population crashes that allow native communities to recover. The abundance and impact of nonnative species may also be closely tied to temporally variable habitat characteristics. We investigated the long-term effects of habitat fragmentation and invasion by the Argentine ant (Linepithema humile) by resurveying ants in 40 scrub habitat fragments in coastal southern California that were originally sampled 21 yr ago. At a landscape scale, fragment area, but not fragment age or Argentine ant mean abundance, continued to explain variation in native ant species richness; the species–area relationship between the two sample years did not differ in terms of slope or intercept. At local scales, over the last 21 yr we detected increases in the overall area invaded (+36.7%, estimated as the proportion of occupied traps) and the relative abundance of the Argentine ant (+121.95%, estimated as mean number of workers in pitfall traps). Argentine ant mean abundance also increased inward from urban edges in 2017 compared to 1996. The greater level of penetration into fragments likely reduced native ant richness by eliminating refugia for native ants in fragments that did not contain sufficient interior area. At one fragment where we sampled eight times over the last 21 yr, Argentine ant mean abundance increased over time while the diversity of native ground-foraging ants declined from 14 to 4 species. Notably, native species predicted to be particularly sensitive to the combined effect of invasion and habitat loss were not detected at any sites in our recent sampling, including the army ant genus Neivamyrmex. Conversely, two introduced ant species (Brachymyrmex patagonicus and Pheidole flavens) that were undetected in 1996 are now common and widespread at our sites. Our results indicate that behaviorally and numerically dominant invasive species can maintain high densities and suppress native diversity for extended periods.

Invasive species threaten biodiversity, ecosystem function, and human health, but the long-term drivers of invasion dynamics remain poorly understood. We use data from a 28-yr ongoing survey of a Northern California ant community invaded by the Argentine ant (Linepithema humile) to investigate the influence of abiotic and biotic factors on invasion dynamics. We found that the distribution of L. humile retracted following an extreme drought that occurred in the region from 2012 to 2015. The distribution of several native ant species also contracted, but overall native ant diversity was higher after the drought and for some native ant species, distributions expanded over the 28-yr survey period. Using structural equation models, we found the strongest impact on the distribution of L. humile was from direct effects of climate, namely, cumulative precipitation and summer maximum temperatures, with only a negligible role for biotic resistance and indirect effects of climate mediated by native ants. The increasing drought and high temperature extremes projected for northern California because of anthropogenic-driven climate change may limit the spread, and possibly the impact, of L. humile in invaded regions. The outcome will depend on the response of native ant communities to these climatic stressors.

A diversity of decision-making systems has been observed in animal collectives. In some species, choices depend on the differences of the numbers of animals that have chosen each of the available options, whereas in other species on the relative differences (a behavior known as Weber’s law), or follow more complex rules. We here show that this diversity of decision systems corresponds to a single rule of decision making in collectives. We first obtained a decision rule based on Bayesian estimation that uses the information provided by the behaviors of the other individuals to improve the estimation of the structure of the world. We then tested this rule in decision experiments using zebrafish (Danio rerio), and in existing rich datasets of argentine ants (Linepithema humile) and sticklebacks (Gasterosteus aculeatus), showing that a unified model across species can quantitatively explain the diversity of decision systems. Further, these results show that the different counting systems used by animals, including humans, can emerge from the common principle of using social information to make good decisions.

The Argentine ant, Linepithema humile (Mayr), is one of the world’s most damaging invasive species. Current control strategies for L. humile rely on neurotoxic insecticides; however, their use is increasingly limited due to their environmental impacts and subsequent regulatory restrictions. Juvenile hormone analogues, such as methoprene, may offer an alternative solution due to their low toxicity to non-target organisms and more favorable environmental profiles. While some juvenile hormone analogues have been tested against several myrmicine ants, their effects on other subfamilies, such as Dolichoderinae, remain understudied. Only one peer-reviewed publication has evaluated methoprene’s effect on Argentine ant colonies in the laboratory, reporting increased mortality in adult workers. However, the study did not explore potential physiological mechanisms underlying this observation. Research findings from other insect taxa suggest that juvenile hormone and their synthetic analogues may disrupt adult physiology by altering lipid metabolism and cuticular hydrocarbon profiles, key traits involved in desiccation resistance and chemical communication. The current study investigated the effects of methoprene on the cuticular hydrocarbon profiles in L. humile . To administer methoprene in a controlled manner, small colony fractions housed in sealed enclosures were exposed to methoprene vapor. After 21 days, cuticular hydrocarbons were extracted from adult workers and queens and quantified using gas chromatography. Methoprene exposure significantly reduced the total cuticular hydrocarbon quantity in both castes. Moreover, the effect of methoprene on CHCs was dependent on their class and chain length, with caste-specific patterns. These findings suggest methoprene disrupts the lipid metabolic processes linked to cuticular hydrocarbon biosynthesis. These findings may provide a foundation to further explore the physiological impacts of methoprene and other juvenile hormone analogues on Argentine ants and other pestiferous dolichoderine ants.