RNA virus discoveries in the electric ant, Wasmannia auropunctata.

Despite being one of the most destructive invasive species of ants, only two natural enemies are known currently for Wasmannia auropunctata, commonly known as the electric ant or little fire ant. Because viruses can be effective biological control agents against many insect pests, including ants, a metagenomics/next-generation sequencing approach was used to facilitate discovery of virus sequences from the transcriptomes of W. auropunctata. Five new and complete positive sense, single-stranded RNA virus genomes, and one new negative sense, single-stranded RNA virus genome were identified, sequenced, and characterized from W. auropunctata collected in Argentina by this approach, including a dicistrovirus (Electric ant dicistrovirus), two polycipiviruses (Electric ant polycipivirus 1; Electric ant polycipivirus 2), a solinvivirus (Electric ant solinvivirus), a divergent genome with similarity to an unclassified group in the Picornavirales (Electric ant virus 1), and a rhabdovirus (Electric ant rhabdovirus). An additional virus genome was detected that is likely Solenopsis invicta virus 10 (MH727527). The virus genome sequences were absent from the transcriptomes of W. auropunctata collected in the USA (Hawaii and Florida). Additional limited field surveys corroborated the absence of these viruses in regions where the electric ant is invasive (the USA and Australia). The replicative genome strand of four of the viruses (Electric ant polycipivirus 2, Electric ant solinvivirus, Electric ant virus 1, and Solenopsis invicta virus 10 (in the electric ant) was detected in Argentinean-collected W. auropunctata indicating that the ant is a host for these viruses. These are the first virus discoveries to be made from W. auropunctata.

Supercolonial structure of invasive populations of the tawny crazy ant Nylanderia fulva in the US.

BACKGROUND: Social insects are among the most serious invasive pests in the world, particularly successful at monopolizing environmental resources to outcompete native species and achieve ecological dominance. The invasive success of some social insects is enhanced by their unicolonial structure, under which the presence of numerous queens and the lack of aggression against non-nestmates allow high worker densities, colony growth, and survival while eliminating intra-specific competition. In this study, we investigated the population genetics, colony structure and levels of aggression in the tawny crazy ant, Nylanderia fulva, which was recently introduced into the United States from South America. RESULTS: We found that this species experienced a genetic bottleneck during its invasion lowering its genetic diversity by 60%. Our results show that the introduction of N. fulva is associated with a shift in colony structure. This species exhibits a multicolonial organization in its native range, with colonies clearly separated from one another, whereas it displays a unicolonial system with no clear boundaries among nests in its invasive range. We uncovered an absence of genetic differentiation among populations across the entire invasive range, and a lack of aggressive behaviors towards conspecifics from different nests, even ones separated by several hundreds of kilometers. CONCLUSIONS: Overall, these results suggest that across its entire invasive range in the U.S.A., this species forms a single supercolony spreading more than 2000 km. In each invasive nest, we found several, up to hundreds, of reproductive queens, each being mated with a single male. The many reproductive queens per nests, together with the free movement of individuals between nests, leads to a relatedness coefficient among nestmate workers close to zero in introduced populations, calling into question the stability of this unicolonial system in which indirect fitness benefits to workers is apparently absent.

Prospecting for viral natural enemies of the fire ant Solenopsis invicta in Argentina.

Metagenomics and next generation sequencing were employed to discover new virus natural enemies of the fire ant, Solenopsis invicta Buren in its native range (i.e., Formosa, Argentina) with the ultimate goal of testing and releasing new viral pathogens into U.S. S. invicta populations to provide natural, sustainable control of this ant. RNA was purified from worker ants from 182 S. invicta colonies, which was pooled into 4 groups according to location. A library was created from each group and sequenced using Illumina Miseq technology. After a series of winnowing methods to remove S. invicta genes, known S. invicta virus genes, and all other non-virus gene sequences, 61,944 unique singletons were identified with virus identity. These were assembled de novo yielding 171 contiguous sequences with significant identity to non-plant virus genes. Fifteen contiguous sequences exhibited very high expression rates and were detected in all four gene libraries. One contig (Contig_29) exhibited the highest expression level overall and across all four gene libraries. Random amplification of cDNA ends analyses expanded this contiguous sequence yielding a complete virus genome, which we have provisionally named Solenopsis invicta virus 5 (SINV-5). SINV-5 is a positive-sense, single-stranded RNA virus with genome characteristics consistent with insect-infecting viruses from the family Dicistroviridae. Moreover, the replicative genome strand of SINV-5 was detected in worker ants indicating that S. invicta serves as host for the virus. Many additional sequences were identified that are likely of viral origin. These sequences await further investigation to determine their origins and relationship with S. invicta. This study expands knowledge of the RNA virome diversity found within S. invicta populations.

Effect of Irradiation on Queen Survivorship and Reproduction in the Invasive Fire Ant Solenopsis invicta (Hymenoptera: Formicidae) and a Proposed Phytosanitary Irradiation Treatment for Ants.

We studied radiation tolerance in queens of the red imported fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) to identify a dose that prevents reproduction. Virgin or fertile queens were collected from Santa Fe and Formosa provinces in Argentina and reared in the laboratory in microcolonies. Virgin queens were irradiated at 0 (control), 70, 90, 120, or 150 Gy, and fertile queens were irradiated at 0, 60, 125, and 190 Gy, and then followed for 11 wk in the microcolonies to evaluate survival and reproduction. Virgin queens lay trophic eggs that do not hatch, whereas fertile queens lay eggs that hatch and develop into brood. In general, queen oviposition and survival decreased with increasing irradiation dose. For virgin queens, no eggs were laid by irradiated queens after the third week, whereas the control queens continued laying eggs throughout the 11-wk experiment. For fertile queens, only one larva and no pupae was observed in the 60 Gy treatment and no larvae or pupae were observed in the 125 and 190 Gy treatments, whereas a total of 431 larvae and 83 pupae were produced by untreated control queens during 11 wks. Survivorship of virgin and fertile queens was similarly reduced by irradiation treatment. These results with S. invicta are consistent with previous findings for three other invasive ants, Wasmannia auropunctata (Roger), Pheidole megacephala (F.), and Linephithema humile (Mayr), that are hitchhiker pests on fresh horticultural commodities. A radiation dose of 150 Gy is proposed as a phytosanitary treatment to prevent reproduction in ants.

Acute exposure to low-dose radiation disrupts reproduction and shortens survival of Wasmannia auropunctata (Hymenoptera: Formicidae) queens.

Irradiation is a postharvest quarantine treatment option to control ants and other hitchhiker pests on fresh horticultural products exported from Hawaii. The radiotolerance of the invasive little fire ant, Wasmannia auropunctata (Roger) (Hymenoptera: Formicidae: Myrmicinae), was studied to determine a dose sufficient for its control. Queens from each of five laboratory microcolonies started from five geographic locations in Argentina were irradiated at 20, 50, 70, or 100 Gy or left untreated as controls and then followed for 13 wk to observe colony growth. In general, queen survivorship, and the number of eggs, larvae, and pupae observed in the microcolonies decreased with increasing irradiation dose. In the 50-, 70-, and 100-Gy treatments, the number of eggs observed was reduced by 68, 66, and 76%, respectively, compared with untreated control microcolonies. The number of larvae in the 50-, 70-, and, 100-Gy treatments was reduced by 99.6%, and only one pupa was observed in the 50-Gy treatment and none in the 70- and 100-Gy treatments during the 13-wk experiment. Queens in the 100-Gy treatment had significantly reduced longevity compared with queens in the other treatments. Radiation doses > or = 70 Gy stopped reproduction in W. auropunctata queens and should be sufficient as a phytosanitary treatment. Information from additional invasive ants in Myrmicinae and other subfamilies is needed before recommending a generic irradiation treatment for ants.

Ecological dominance of the red imported fire ant, Solenopsis invicta, in its native range.

Despite the widespread impacts invasive species can have in introduced populations, little is known about competitive mechanisms and dominance hierarchies between invaders and similar taxa in their native range. This study examines interactions between the red imported fire ant, Solenopsis invicta, and other above-ground foraging ants in two habitats in northeastern Argentina. A combination of pitfall traps and baits was used to characterize the ant communities, their dominance relationships, and to evaluate the effect of phorid flies on the interactions. Twenty-eight ant species coexisted with S. invicta in a gallery forest gap, whereas only ten coexisted with S. invicta in a xerophytic forest grassland. S. invicta was the most numerically dominant species in the richest and complex habitat (gallery forest); however it performed better as discoverer and dominator in the simpler habitat. S. invicta was active during day and night. In spite of its poor capacity to discover resources, S. invicta showed the highest ecological dominance and the second-best behavioral dominance after Camponotus blandus. S. invicta won 78% of the interactions with other ants, mostly against its most frequent competitor, Pheidole cf. obscurithorax, dominating baits via mass recruitment and chemical aggression. P. cf. obscurithorax was the best food discoverer. S. invicta won 80% of the scarce interactions with Linepithema humile. Crematogaster quadriformis was one of the fastest foragers and the only ant that won an equal number of contests against S. invicta. The low presence of phorid flies affected the foraging rate of S. invicta, but not the outcome of interspecific interactions. This study revealed that the red imported fire ant ecologically dominated other terrestrial ants in its native range; however, other species were able to be numerically dominant or co-dominant in its presence.

Phenology, distribution, and host specificity of Solenopsis invicta virus-1.

Studies were conducted to examine the phenology, geographic distribution, and host specificity of the Solenopsis invicta virus-1 (SINV-1). Two genotypes examined, SINV-1 and -1A, exhibited similar seasonal prevalence patterns. Infection rates among colonies of S. invicta in Gainesville, Florida, were lowest from early winter (December) to early spring (April) increasing rapidly in late spring (May) and remaining high through August before declining again in the fall (September/October). Correlation analysis revealed a significant relationship between mean monthly temperature and SINV-1 (p<0.0005, r=0.82) and SINV-1A (p<0.0001, r=0.86) infection rates in S. invicta colonies. SINV-1 was widely distributed among S. invicta populations. The virus was detected in S. invicta from Argentina and from all U.S. states examined, with the exception of New Mexico. SINV-1 and -1A were also detected in other Solenopsis species. SINV-1 was detected in Solenopsis richteri and the S. invicta/richteri hybrid collected from northern Alabama and Solenopsis geminata from Florida. SINV-1A was detected in S. geminata and Solenopsis carolinensis in Florida and the S. invicta/richteri hybrid in Alabama. Of the 1989 arthropods collected from 6 pitfall trap experiments from Gainesville and Williston, Florida, none except S. invicta tested positive for SINV-1 or SINV-1A. SINV-1 did not appear to infect or replicate within Sf9 or Dm-2 cells in vitro. The number of SINV-1 genome copies did not significantly increase over the course of the experiment, nor were any cytopathic effects observed. Phylogenetic analyses of SINV-1/-1A nucleotide sequences indicated significant divergence between viruses collected from Argentina and the U.S.