Interactions between sympatric invasive European fire ants (Myrmica rubra) and blacklegged ticks (Ixodes scapularis)

The blacklegged tick (Ixodes scapularis) and the invasive European fire ant (Myrmica rubra) are both expanding throughout their sympatric range in coastal New England. Ixodes scapularis is the primary vector of the bacterium Borrelia burgdorferi, which is the causative agent of Lyme disease, and Mount Desert Island, Maine, home to Acadia National Park, currently is affected by a high Lyme disease burden. Ticks have many natural predators, including ants, although no previous studies have investigated interactions between these two species. To test the hypothesis that the presence of M. rubra alters I. scapularis abundance, we collected ticks by drag-sampling at eight ant-infested sites and eight uninfested control sites in Acadia National Park. We found that nymph density was significantly higher at ant-infested sites, while larval density was significantly higher at control sites. In addition, we conducted a laboratory bioassay to measure M. rubra aggression against I. scapularis larvae, nymphs, and adults and Dermacentor variabilis adults, and found that ant aggression was significantly higher against D. variabilis adults than I. scapularis adults. Our findings support the hypothesis that M. rubra has divergent effects across I. scapularis life stages, and we discuss possible ecological mechanisms, including optimal microclimate and predation, that could promote density of nymphs while inhibiting density of larvae.

Assessment of Pathogenic Bacteria Transfer From Pristionchus Entomophagus (Nematoda: Diplogasteridae) to the Invasive Ant Myrmica Rubra and Its Potential Role in Colony Mortality in Coastal Maine

Background:Necromenic nematode Pristionchus entomophagus has been frequently found in nests of the invasive European ant Myrmica rubra in coastal Maine, United States. The nematodes may contribute to ant mortality and collapse of colonies by transferring environmental bacteria. M. rubra ants naturally hosting nematodes were collected from collapsed wild nests in Maine and used for bacteria identification. Virulence assays were carried out to validate acquisition and vectoring of environmental bacteria to the ants.Results:Multiple bacteria species, including Paenibacillus spp., were found in the nematodes’ digestive tract. Serratia marcescens, Serratia nematodiphila, and Pseudomonas fluorescens were collected from the hemolymph of nematode-infected Galleria mellonella larvae. Variability was observed in insect virulence in relation to the site origin of the nematodes. In vitro assays confirmed uptake of RFP-labeled Pseudomonas aeruginosa strain PA14 by nematodes. Bacteria were highly concentrated in the digestive tract of adult nematodes, a small amount of bacteria were observed in the digestive tract of juveniles with a more significant amount on their cuticle, and none on the cuticle of adults. RFP-labeled P. aeruginosa were not observed in hemolymph of G. mellonella larvae, indicating an apparent lack of bacterial transfer from juvenile nematodes to the insects despite larval mortality.Host species was the primary factor affecting bacterial community profiles. Spiroplasma sp. and Serratia marcescens sequences were shared across ants, nematodes, and nematode-exposed G. mellonella larvae. Alternative to the idea of transferring bacteria from environment to host, we considered whether nematode-exposure might disorder or depauperate the endobiotic community of an insect host. While total bacterial diversity was not statistically lower in nematode-exposed G. mellonella larvae when compared to controls, 16 bacterial sequence variants were less abundant in nematode-exposed larvae, while three were increased, including Serratia, Pseudomonas, and Proteus.Conclusions: This study suggests that transfer of bacteria from nematodes to ants is feasible, although largely serendipitous, and may possibly contributed to ant death and/or collapse of wild colonies in Maine. Hypothetically, the use of an engineered biological control, such as nematodes carrying specifically-seeded bacterial species, may be effective, especially if the pathogenic bacteria are normally found in soil ecosystems and represents a low risk for biosafety control.

Prophylactic Avoidance of Hazardous Prey by the Ant Host Myrmica rubra

Ants are the hosts of many microorganisms, including pathogens that are incidentally brought inside the nest by foragers. This is particularly true for scavenging species, which collect hazardous food such as dead insects. Foragers limit sanitary risks by not retrieving highly infectious prey releasing entomopathogenic fungal spores. This study investigates whether similar prophylactic strategies are also developed for food associated with weak or delayed risks of fungal contamination. We compared, in Myrmica rubra ant colonies, the retrieval dynamics of dead flies that were (1) conidia-free, (2) covered with a low amount of Metarhizium brunneum entomopathogenic conidia or (3) recently fungus-killed but not yet sporulating. Foragers mostly avoided fungus-killed prey and delayed the retrieval of conidia-covered flies. A second sanitary filter occurred inside the nest through a careful inspection of the retrieved prey. Ultimately, ants mostly consumed conidia-free and conidia-covered flies, but they relocated and discarded all fungus-killed prey outside of the nest. Our study confirms that, as a host of generalist entomopathogenic fungi, Myrmica rubra ants have developed a prophylactic avoidance and a differential management of prey depending on their infectious potential. We discuss the functional value as well as the possible cues underlying pathogen avoidance and prey discrimination in ants.

Nest Entrances, Spatial Fidelity, and Foraging Patterns in the Red Ant Myrmica rubra: A Field and Theoretical Study

The nest architecture of social insects deeply impacts the spatial distribution of nestmates their interactions, information exchanges and collective responses. In particular, the number of nest entrances can influence the interactions taking place beyond the nest boundaries and the emergence of collective structures like foraging trails. Here, we investigated in the field how the number of nest entrances impacted the foraging dynamics of Myrmica rubra ant colonies. We located the nest entrances where recruitment occurred towards sugar feeders placed in their surroundings. The nests showed one or multiple entrance(s) aggregated in clusters spaced by at least 15 cm. Foragers from colonies with two clusters of entrances were distributed more homogeneously among the feeders than those of colonies with one cluster. In addition, foragers always returned to the first discovered feeder and demonstrated a high fidelity to their original entrance. Finally, a multi-agent model highlighted that additional entrances and clusters of entrances delayed the mobilisation of workers but favoured the simultaneous exploitation of several sources, which was further enhanced by the spatial fidelity of foragers. Multiple nest entrances seem to be a way for medium-sized colonies to benefit from advantages conferred by polydomy while avoiding associated costs to maintain social cohesion.

Evaluation of Four Plant Extract Repellents for Management of the European Red Ant Myrmica rubra (Hymenoptera: Formicidae)

Spearmint oil, peppermint oil, neem oil, and d-limonene were tested as nest site repellents against the colonization of the invasive European red ant, Myrmica rubra (L.) in both laboratory and field trials. In a laboratory assay, a 10% (v/v) solution of each extract repelled M. rubra colonies from nesting in plant pots filled with moist soil compared to water-treated controls, when applied as a dip to pots. Extracts also repelled colonies compared to a water control in a second laboratory experiment, where pots were dipped 15 d prior to the start of the experiment. In a 2008 field comparison, 20 plant pots, filled with moist potting soil, were dipped in either 10% (v/v) spearmint oil or peppermint oil, 70% (v/v) neem oil, or a water control and left in infested sites. Seven control pots were colonized over 3 mo, while none of the extract-treated pots were colonized. This field trial was repeated for a 15-wk duration in 2009 with the addition of a 10% (v/v) d-limonene solution as a treatment, and a change in neem oil concentration to 10% (v/v). Spearmint and peppermint oils repelled M. rubra colonies for the duration of the experiment. Neem oil and d-limonene repelled colonies for 3 and 4 wk, respectively. These extracts, especially mint oils, show potential as low-hazard repellents against M. rubra in greenhouse and nursery settings, and could reduce the number of new infestations incurred by the transport of plant stock.

Pathogen avoidance and prey discrimination in ants

Insect societies have developed sanitary strategies, one of which is the avoidance of infectious food resources as a primary line of defence. Using binary choices, we investigated whether Myrmica rubra ants can identify prey that has been artificially infected with the entomopathogenic fungus, Metarhizium brunneum . We compared the ants' foraging behaviour towards infected prey at three different stages of fungus development : (i) prey covered with fungal conidia, (ii) prey freshly killed by the fungus and (iii) sporulating prey. Most foragers retrieved a corpse covered with a high number of spores but they consistently avoided a sporulating prey and collected less prey that had recently died from fungal infection. Furthermore, ant responses were highly variable, with some individuals retrieving the first prey they encountered while others inspected both available prey before making a decision. Workers were not repelled by the simple presence of fungal conidia but nevertheless, they avoided retrieving cadavers at later stages of fungal infection. We discuss how these different avoidance responses could be related to: differences in the ants’ perceptive abilities; physico-chemical cues characterizing fungus-infected prey or in the existence of physiological or behavioural defences that limit sanitary risks associated with potentially contaminated resources.

Foodborne Transmission of Deformed Wing Virus to Ants (Myrmica rubra)

Virus host shifts occur frequently, but the whole range of host species and the actual transmission pathways are often poorly understood. Deformed wing virus (DWV), an RNA virus described from honeybees (Apis mellifera), has been shown to have a broad host range. Since ants are often scavenging on dead honeybees, foodborne transmission of these viruses may occur. However, the role of the ant Myrmica rubra as an alternative host is not known and foodborne transmission to ants has not been experimentally addressed yet. Here, we show with a 16-week feeding experiment that foodborne transmission enables DWV type-A and -B to infect M. rubra and that these ants may serve as a virus reservoir. However, the titers of both plus- and minus-sense viral RNA strands decreased over time. Since the ants were fed with highly virus-saturated honeybee pupae, this probably resulted in initial viral peaks, then approaching lower equilibrium titers in infected individuals later. Since DWV infections were also found in untreated field-collected M. rubra colonies, our results support the wide host range of DWV and further suggest foodborne transmission as a so far underestimated spread mechanism.