Host preference in Pseudacteon phorid flies: response of P. tricuspis and P. curvatus to black, red and hybrid imported Solenopsis fire ants in multiple choice bioassays

Host preferences in both sexes of Pseudacteon tricuspis Borgmeier (Jaguariuna biotype) and Pseudacteon curvatus Borgmeier (Formosa biotype) and their relative attraction to the imported fire ants (IFA), Solenopsis invicta Buren (red IFA), Solenopsis richteri Forel (black IFA) and S. invicta × S. richteri hybrids (hybrid IFA) were investigated in two separate experiments utilizing multiple choice flight bioassays. The results of both experiments clearly showed that both sexes of the Jaguariuna biotype of P. tricuspis could distinguish among the three IFA species and demonstrated greater preference for hybrid IFA and red IFA. This conclusion is supported by a variety of data collected on the number of fly visits, attack rate, and hovering duration (Experiment 1), and on the number of trapped flies (Experiment 2), which showed that black IFA is the least preferred of the three species. Similar results were recorded for the Formosan biotype of P. curvatus, although the data were not as strongly conclusive. Females of this biotype spent a significantly greater amount of time in hovering mode over red IFA and hybrid IFA compared to black IFA, but the other data were not significant. The red IFA is the natural host of both phorid fly biotypes and our results suggest that both biotypes may have evolved a specialized relationship with red IFA including an ability to discriminate it from related fire ants. These results are discussed in relation to the possible role of fire ant chemicals in mediating host preferences in phorid flies, contributions of male phorid flies to fire ant biocontrol, and the practical implications of the key findings.     

Zombie fire ant workers: behavior controlled by decapitating fly parasitoids

Laboratory observations were conducted on four separate red imported fire ant, Solenopsis invicta, colonies that contained workers parasitized by the decapitating fly, Pseudacteon tricuspis. Parasitized S. invicta workers remained inside the nest during parasitoid larval development and left the nest approximately 8 – 10 hours before decapitation by the parasitoid. When parasitized ants left the nest, they were highly mobile, were responsive to tactile stimuli, and showed minimal defensive behavior. Ants ultimately entered into a grass thatch layer, where they were decapitated and the fly maggots pupariated. This study reveals that parasitized ants exhibit behaviors that are consistent with host manipulation to benefit survival of the parasitoid. Received 9 November 2006; revised 26 January 2007; accepted 7 February.     

Isolation, characterization, and molecular identification of bacteria from the red imported fire ant (Solenopsis invicta) midgut

Bacteria were isolated and cultured from the red imported fire ant (Solenopsis invicta) midgut. The small-subunit ribosomal RNA gene, (16s rRNA gene, approximately 1500 bp) was amplified from bacterial genomic DNA using the polymerase chain reaction and consensus sequence primers. Restriction fragment length polymorphism analysis revealed 10 unique profiles, indicating that at least 10 different bacteria are present in red imported fire ant midguts. The 16s rRNA gene sequence was determined for these isolates and queried against the NCBI genetic database. The results identified all isolates to at least the genus level. Antibiotic resistance profiles and biochemical activities were also determined for these species. This work provides the basis for a wider characterization of bacterial distributions in fire ant colonies and provides strains suitable for genetic manipulation to develop novel methods of fire ant control.     

Behavioral and Electroantennogram Responses of Phorid fly <Emphasis Type="Italic">Pseudacteon tricuspis</Emphasis> (Diptera: Phoridae) to Red Imported Fire Ant <Emphasis Type="Italic">Solenopsis invicta</Emphasis> Odor and Trail Pheromone

The phorid fly, Pseudacteon tricuspis is an introduced parasitoid of the red imported fire ant, Solenopsis invicta in the United States. Previous studies show that phorid flies are attracted to host ant workers at disturbed colonies, to colonies exhibiting aggressive interspecific interactions, and to fire ant mating flights. In a series of behavioral and electroantennogram (EAG) experiments, we confirm the possible use of fire ant odor as cues for host location by P. tricuspis. We tested the response of P. tricuspis of different sex and mating status to several host-related odor stimuli including live fire ant workers, extracts of worker whole body, head, thorax, and abdomen, and (E,E)-α-farnesene, a trail pheromone component of Solenopsis fire ants. Results from Y-tube olfactometer bioassays demonstrated the attraction of mated female P. tricuspis to live S. invicta workers. In addition, extracts of S. invicta worker whole body and thorax elicited strong olfactometer response in female flies (mated and unmated) and mated males, but not in unmated males. Pseudacteon tricuspis did not show significant attraction to extracts of S. invicta worker head and abdomen, or to (E,E)-α-farnesene, irrespective of sex and mating status. In EAG experiments, female and male P. tricuspis showed significant EAG response to extracts of worker whole body, head, and abdomen, and to a less extent, thorax extract, but not to (E,E)-α-farnesene. Females showed slightly greater EAG response than males, but EAG response was not affected by mating status. These results suggest that fire ant thorax is likely the source of kairomones used as host location cues by P. tricuspis, and support the hypothesis that fire ant worker trail pheromones are not likely used by P. tricuspis for host location.     

Pseudacteon decapitating flies (Diptera: Phoridae): Are they potential vectors of the fire ant pathogens Kneallhazia (=Thelohania) solenopsae (Microsporidia: Thelohaniidae) and Vairimorpha invictae (Microsporidia: Burenellidae)?

Fire ant decapitating flies in the genus Pseudacteon were tested for their potential as hosts or vectors of two microsporidian pathogens of the red imported fire ant, Solenopsis invicta. Decapitating flies that attacked or were reared from S. invicta workers infected by Kneallhazia (=Thelohania) solenopsae or Vairimorpha invictae were tested for either pathogen by polymerase chain reaction (PCR) tests or visual examination for spores using phase microscopy. Three species of fire ant decapitating flies acquired the pathogen, Kneallhazia solenopsae. K. solenopsae was detected in 58% of pooled samples of Pseudacteon obtusus flies and 44% of pooled samples of Pseudacteon cultellatus that developed in K. solenopsae-infected fire ant workers. K. solenopsae was also found in 17% of pooled samples of field-collected Pseudacteon curvatus. In contrast, the microsporidium V. invictae was not detected in P. obtusus reared from V. invictae-infected S. invicta workers. Neither K. solenopsae nor V. invictae were detected in any of the hovering or ovipositing flies in the laboratory exposures, indicating no mechanical acquisition of the microsporidia occurred during oviposition activity. Greater than 92% of the P. obtusus that developed in K. solenopsae-infected ants survived and emerged as adults, thus indicating no detrimental effects of the microsporidium on pupal development and emergence. These results indicate that Pseudacteon decapitating flies may be able to vector K. solenopsae but not V. invictae among fire ants. Further tests are planned to determine if flies containing K. solenopsae are capable of transferring this pathogen either during oviposition or by being consumed by fire ant larvae.     

Impacts of Argentine ants on avian nesting success

Biological invasions can have severe and widespread impacts on ecological communities. A few species of ants have become particularly damaging invaders but quantitative data of their impacts on many taxa is still lacking. We provide experimental evidence using artificial nests baited with quail eggs that the invasive Argentine ant (Linepithema humile) can be a significant avian nest predator – Argentine ants recruited to more nests and in higher abundance than the native ant species they displace. However, at a site invaded by Argentine ants, we monitored over 400 nests of a ground-nesting species, the Dark-eyed Junco (Junco hyemalis), and found that less than 2% of nests failed as a result of Argentine ant predation/infestation. A review of the literature also suggests that Argentine ants may not be a serious threat to bird nests relative to other predators or parasites. However, invasive ants with the capability of overwhelming prey though stinging (specifically the red-imported fire ant, Solenopsis invicta), may have a higher impact on avian nesting success. Received 14 January 2005; revised 28 April 2005; accepted 12 May 2005.     

Effect of mono- and polygyne social forms on transmission and spread of a microsporidium in fire ant populations

Thelohania solenopsae is a pathogen of the red imported fire ant, Solenopsis invicta, which debilitates queens and eventually causes the demise of colonies. Reductions of infected field populations signify its potential usefulness as a biological control agent. Thelohania solenopsae can be transmitted by introducing infected brood into a colony. The social forms of the fire ant, that is, monogyny (single queen per colony) or polygyny (multiple queens per colony), are associated with different behaviors, such as territoriality, that affect the degree of intercolony brood transfer. T. solenopsae was found exclusively in polygyne colonies in Florida. Non-synchronous infections of queens and transovarial transmission favor the persistence and probability of detecting infections in polygynous colonies. However, queens or alates with the monogyne genotype can be infected, and infections in monogyne field colonies have been reported from Louisiana and Argentina. Limited independent colony-founding capability and shorter dispersal of alate queens with the polygyne genotype relative to monogyne alates may facilitate the maintenance of infections in local polygynous populations. Demise of infected monogyne colonies can be twice as fast as in polygyne colonies and favors the pathogen's persistence in polygyne fire ant populations. The social form of the fire ant reflects different physiological and behavioral aspects of the queen and colony that will impact T. solenopsae spread and ultimate usefulness for biological control.     

Culture-independent identification of gut bacteria in fourth-instar red imported fire ant, Solenopsis invicta Buren, larvae

Red imported fire ants (RIFA), Solenopsis invicta Buren, are medical, urban, and agricultural pests from South America. They are successful invaders due to their preference for disturbed habitats, high reproductive rates, and the ability to feed on a wide variety of food items (omnivorous). Fourth-instar larvae are used by the colony to digest solid food and then regurgitate it for consumption by workers and queens. Larvae are an ideal source of investigations of endosymbiotic bacteria possibly involved in nutrient distributions. Our study utilized 16S rDNA sequencing to describe the composition of the bacterial community in fourth-instar ant larvae in order to identify possible endosymbiotic bacteria present therein. The 16S rRNA gene was directly amplified from mixed-population DNA of whole fire ant larval guts and cloned into Escherichia coli. Bacterial communities from three geographically separated RIFA colonies were examined. Sequenced bacterial clones from guts were determined to be predominantly from the phylum Proteobacteria and the family Enterobacteriaceae. Our results did not detect the presence of endosymbiotic bacteria in the guts of RIFA larvae among the colonies. In addition, minimal species overlap was found when bacterial inventories were compared among colonies. Thus, bacteria coadapted with red imported fire ant larvae were not detected. Identified bacteria were not closely affiliated with endosymbiotic bacteria common in other insect species. Bacteria communities appeared to be unique to each geographical location and were determined by the foods consumed by the ants.     

Dynamic expansion in recently introduced populations of fire ant parasitoids (Diptera: Phoridae)

Combating invasive species requires a detailed, mechanistic understanding of the manner and speed with which organisms expand their ranges. Biological control efforts provide an opportunity to study the process of species invasions and range expansions under known initial conditions. This study examines the rate, pattern and mechanisms of spread for two populations of the biological control agent Pseudacteon tricuspis, phorid-fly parasitoids of imported fire ants. We employ a trap-based survey method that detects phorid flies in low-density populations, and provides data on abundance. This technique allows us to differentiate between continuous population spread and effective long-distance dispersal and to examine density gradients of phorid flies across the expanding population front. We find that occupied sites in front of the leading edge of continuous populations were common; forming small populations we refer to as satellite populations. Satellite populations are tens of kilometers from the nearest possible source. Wind governs the dynamics of spread in these two central Texas populations. Population edges expanding with the wind exhibited a higher frequency of effective long-distance dispersal than did populations expanding into the wind. This enhanced effective long-distance dispersal rate translated into a five times faster rate of spread for population edges traveling with the wind. This planned invasion shares many characteristics in common with unplanned species invasions including: protracted establishment phase during which densities were below detection thresholds, and slow initial spread immediately after establishment followed by rapid, accelerating spread rates as population sizes grew.     

Post-Release Host-Specificity Testing of Pseudacteon tricuspis, a Phorid Parasitoid of Solenopsis Invicta Fire Ants

Inherent in any biological control program is the risk of nontarget effects. Pseudacteon tricuspisBorgmeier, a parasitoid phorid fly, has been introduced to the United States from South America as a potential biocontrol agent of the red imported fire ant, Solenopsis invictaBuren. We conducted tests of host specificity on introduced populations of P. tricuspis, which are attracted to alarm pheromones released by their hosts during events such as mound disturbances and interspecific interactions. We monitored disturbed mounds of S. invicta and its close congener, S. geminata(F.), during the expansion of P. tricuspis across north Florida and after populations had been established for ~3 years. We also tested host acceptance in established populations of P. tricuspis by offering trays containing S. invicta, S. geminata, and 14 additional ant species representing 12 different non-Solenopsis genera. Although P. tricuspiswas commonly observed to hover over and attempt to oviposit on S. invicta, we never observed any parasitization attempts on any other ant species. As predicted by laboratory tests, released populations of P. tricuspis appear to be highly host specific and pose no obvious threat to nontarget species.     

The urban fire ant paradox: native fire ants persist in an urban refuge while invasive fire ants dominate natural habitats

In contrast to the widespread extirpation of native fire ants (Solenopsis geminata) across southern US following the invasion by imported red fire ants (S. invicta), some residential areas of Austin form unexpected refuges for native fire ants. Ironically, these urban environments provide refuges for the native fire ants while adjacent natural habitats have been overrun by invasive fire ants. Resistance to invasive fire ants in these urban areas occurs mainly in older residential properties constructed prior to the S. invicta invasion, while more recent construction has allowed establishment by S. invicta. The invasive ability of S. invicta is often attributed to escape from parasitoids and efficient dispersal of polygyne multiple queen colonies. Here we also show the importance of landscape parameters in the invasion process, where low levels of disturbance and continuous plant cover in older residential areas form possible barriers to colonization. Dense leaf cover (high NDVI) was also found to be associated with native ant refuges. Long term residential land ownership may have resulted in lower recent disturbance levels and increased plant cover that support refuges of native fire ants.     

几种杀虫剂对热带雨林生态区红火蚁蚁巢和工蚁的灭杀效果

为筛选出适用于云南热带雨林生态区的红火蚁Solenopsis invicta Buren防治药剂,通过7种杀虫剂对红火蚁蚁巢田间灭除效果试验测定。结果表明,20%多杀霉素悬浮剂（SC）、30%螺虫·噻虫嗪悬浮剂（SC）、70%吡虫啉可湿性粉剂（WP）、10%溴虫氟苯双酰胺悬浮剂（SC）和1%联苯·噻虫嗪颗粒剂（GR）对红火蚁蚁巢处理效果较好,药后1 d和10 d虫口减退率分别在95%和90%以上。其中10%溴虫氟苯双酰胺SC和1%联苯·噻虫嗪GR可作为红火蚁巢点处置的首选药剂;0.05%茚虫威饵剂（EB）速效性差,但药后10 d虫口减退率上升到86.8%。1.8%阿维菌素乳油（EC）药后1 d和10 d虫口减退率分别为89.3%和81%。这2种药剂可与前5种药剂交替使用。     

Parasitoid–host relationships when host size varies: the case of Pseudacteon flies and Solenopsis fire ants

Abstract. 1. Phorid flies in the genus Pseudacteon are parasitoids of ants. Variation in host size preferences of four South American and two North American Pseudacteon species on monogyne and polygyne forms of their host Solenopsis species [ S. invicta Buren and S. geminata (F.), respectively] was documented.
2. Monogyne Solenopsis workers were, on average, significantly larger than polygyne workers, and the average size monogyne worker attacked was significantly larger than the average size polygyne worker attacked in four of the six Pseudacteon species.
3. Three South American Pseudacteon species attacked larger than average size workers, whereas one attacked smaller than average size workers, in both monogyne and polygyne forms. Both North American Pseudacteon species attacked larger than average size polygyne workers and smaller than average size monogyne workers.
4. Three Pseudacteon species were reared from eggs to adults in infected ants in the laboratory. The size of the emergent phorid fly was related positively to the size of the host worker ant, with females emerging from larger hosts. Similar patterns were documented for both monogyne and polygyne forms.
5. The mean size of worker host from which phorids emerged did not differ significantly between the monogyne and polygyne forms in the subsample of phorids reared to adults.
6. The observed patterns elucidate factors that may cause variation in Pseudacteon sex ratios, and have implications for biological control efforts of pest Solenopsis species.     

Ecological interactions of Pseudacteon parasitoids and Solenopsis ant hosts: environmental correlates of activity and effects on competitive hierarchies

1. Solenopsis (Hymenoptera: Formicidae) fire ants are host to Pseudacteon (Diptera: Phoridae) parasitoids. The activity of S. geminata (F.) hosts and relative abundance of Pseudacteon phorids, along with five environmental variables, were measured at weekly intervals over an 8‐month period at two sites. 2. Pseudacteon relative abundances often varied greatly from week to week, and were only weakly positively correlated with S. geminata activity. 3. A quadratic function of soil temperature was the single best predictor of ant activity at both sites, explaining 32 and 73% of the variation in ant activity. A linear function of soil moisture was the single best predictor of phorid relative abundance at one site (r² = 0.23) whereas no measured variables were significant predictors of phorid relative abundance at the other site. 4. Interspecific interactions at 600 baits were monitored at a third site to document dominance hierarchies and determine whether the presence of Pseudacteon phorids mediated interspecific interactions in their host, S. geminata. 5. Solenopsis geminata was near the top of dominance hierarchies, which did not diverge greatly from a linear pattern. Three species (S. geminata, S. invicta Buren, and Crematogaster laeviuscula Mayr) won the majority of their interspecific interactions and appear to be co‐dominants at this microhabitat‐rich site. 6. Overall, the presence of phorids had no significant effect on the outcome of interspecific contests involving S. geminata and all other ant species grouped together. Phorids may have contributed to some of the S. geminata losses against other co‐dominant species.     

Rodents balancing a variety of risks: invasive fire ants and indirect and direct indicators of predation risk

We used foraging trays to compare how oldfield mice, Peromyscus polionotus, altered foraging in response to the presence of fire ants, Solenopsis invicta, and in the presence of direct (predator urine) and indirect (sheltered or exposed microhabitat, moonlight, and precipitation) indicators of predation risk. Foraging reductions elicited by S. invicta were greater than reductions in response to well-documented indicators of risk (i.e., moonlit nights) and the presence of predator urine. The presence of S. invicta always led to reduced foraging, but the overall impact of S. invicta was dependent upon microhabitat and precipitation. When S. invicta was not present, foraging was greater in sheltered microhabitats compared to exposed microhabitats. S. invicta made sheltered microhabitats equivalent to more risky exposed microhabitats, and this effect was especially pronounced on nights without precipitation. The effect of S. invicta suggests that interactions with S. invicta may entail a potentially heavy cost or that presence of S. invicta may represent a more reliable indicator of imminent competition or predation compared to indirect cues of risk and predator urine. The presence of S. invicta led to reduced foraging under situations when foraging activity would otherwise be greatest (i.e., under vegetative cover), potentially reducing habitat quality for P. polionotus and the distribution of seeds consumed by rodents.     

不同浓度茚虫威对红火蚁的防治效果

为探索适合防治红火蚁的茚虫威浓度,利用丙酮溶解茚虫威原药,使用雾化手段与载体和配方其他试剂充分混合,干燥后保存,进行野外试验测定其防治红火蚁的药效。结果表明,研制的0.025%茚虫威饵剂综合防治效果达85.30%,0.05%茚虫威饵剂达100%,0.1%茚虫威饵剂达100%。3个浓度的茚虫威饵剂拥有良好的防治红火蚁药效,0.05%浓度最适合研制红火蚁饵剂。     

Transmission of Vairimorpha invictae (Microsporidia: Burenellidae) infections between red imported fire ant (Hymenoptera: Formicidae) colonies

Red imported fire ant, Solenopsis invicta, colonies were successfully infected with the microsporidium Vairimorpha invictae by introducing live larvae, pupae, or dead adults from V. invictae-infected field colonies collected in Argentina. Introductions with 4th instar larvae or non-melanized pupae obtained from infected field colonies, resulted in infection of 40% of the inoculated colonies. Introductions of 4th instars or melanized pupae produced from colonies that were initially infected in the laboratory, resulted in infections of 83% of the colonies, thus perpetuating the infection in other colonies. Infection was detected in 2 of 6 colonies after introducing adult worker caste ants that had died with V. invictae. The average number of adults and the volume of immature ants per colony were significantly lower in the infected than in the control colonies. Infected colonies had 86% fewer adults per colony and 82% less immature ants than the controls. A portion of the 16S rRNA gene of the V. invictae identified from these studies was amplified, cloned, and sequenced; the 1251 nucleotide amplicon was 100% identical to the 16S rRNA gene sequence recorded previously in the GenBank database, thus verifying the species as V. invictae. This is the first report of the artificial transmission of this pathogen to uninfected ant colonies, and demonstration of its ability to hinder growth in individual colonies.     

Sources of spores for the possible horizontal transmission of Thelohania solenopsae (Microspora: Thelohaniidae) in the red imported fire ants, Solenopsis invicta

We screened adult and larval secretions and midden piles for the presence of Thelohania solenopsae spores to decipher potential sources for the horizontal transmission of the pathogen in fire ants. Hemolymph samples from both adult and larvae were also screened to rule out hemolymph contamination of samples. In adults, Thelohania spores were found in the crop and the fecal fluids, although only free spores were found in the fecal fluids of adults. In fourth instar larvae, both free and octospores were seen in midgut and the meconium samples. All of the midden pile samples had T. solenopsae spores of both types. Based on these results, we theorize that the pathogen may be horizontally transmitted within a colony by the removal and sharing of meconium of prepupating fourth instar larvae by adult workers and by the adult fecal droppings, and intercolonially by contamination of midden piles or brood raiding.