Symbiotic microbiota may reflect host adaptation by resident to invasive ant species.

Exotic invasive species can influence the behavior and ecology of native and resident species, but these changes are often overlooked. Here we hypothesize that the ghost ant, Tapinoma melanocephalum, living in areas that have been invaded by the red imported fire ant, Solenopsis invicta, displays behavioral differences to interspecific competition that are reflected in both its trophic position and symbiotic microbiota. We demonstrate that T. melanocephalum workers from S. invicta invaded areas are less aggressive towards workers of S. invicta than those inhabiting non-invaded areas. Nitrogen isotope analyses reveal that colonies of T. melanocephalum have protein-rich diets in S. invicta invaded areas compared with the carbohydrate-rich diets of colonies living in non-invaded areas. Analysis of microbiota isolated from gut tissue shows that T. melanocephalum workers from S. invicta invaded areas also have different bacterial communities, including a higher abundance of Wolbachia that may play a role in vitamin B provisioning. In contrast, the microbiota of workers of T. melanocephalum from S. invicta-free areas are dominated by bacteria from the orders Bacillales, Lactobacillales and Enterobacteriales that may be involved in sugar metabolism. We further demonstrate experimentally that the composition and structure of the bacterial symbiont communities as well as the prevalence of vitamin B in T. melanocephalum workers from S. invicta invaded and non-invaded areas can be altered if T. melanocephalum workers are supplied with either protein-rich or carbohydrate-rich food. Our results support the hypothesis that bacterial symbiont communities can help hosts by buffering behavioral changes caused by interspecies competition as a consequence of biological invasions.

Transmission modes of a pesticide-degrading symbiont of the oriental fruit fly Bactrocera dorsalis (Hendel).

Symbionts are associated with many insects and play several multifunctional roles in insect-microorganism mutualistic relationships. The trichlorphon-degrading symbiont Citrobacter freundii (CF-BD) of the oriental fruit fly Bactrocera dorsalis was recently discovered; however, its intraspecies transmission pathway among flies remains unknown. Here, we use fluorescence in situ hybridization (FISH), PCR detection, and a series of ingenious experiments to reveal that CF-BD was aggregated in rectal pads associated with the female ovipositor, and the CF-BD symbiont was vertically transmitted via egg surface contamination. Although CF-BD was not detected in ovaries, it was found in deposited eggs. In addition, CF-BD was readily acquired horizontally between larvae or adults via oral uptake, although it was not transferred via mating behavior. Surface sterilization of eggs had a negative effect on the insects, which exhibited a lower body weight and a sharp decrease in fecundity, suggesting important biological roles of CF-BD in the fitness of the host insects. Our findings may also help to explain the high pesticide resistance levels of B. dorsalis. Furthermore, identifying a clear transmission pathway of this organophosphorus-degrading symbiont will be useful for pesticide resistance management and future pest control technologies.

Host Plant-Mediated Interactions Between 'Candidatus Liberibacter asiaticus' and Its Vector Diaphorina citri Kuwayama (Hemiptera: Liviidae).

The Diaphorina citri Kuwayama (Hemiptera: Liviidae) also known as Asian citrus psyllid transmits 'Candidatus Liberibacter asiaticus' (CLas) associated with the citrus Huanglongbing (HLB). Minimizing Asian citrus psyllid populations is one of the most important methods for HLB control. The development and survival of Asian citrus psyllid were evaluated on CLas+ and CLas- plants of Citrus reticulata Blanco. cv. Mashuiju (Sapindales: Rutaceae) or Citrus sunki Hort. ex Tanaka (Sapindales: Rutaceae) to assess the effects of CLas infection on vector populations. The development times were unaffected between CLas+ and CLas- plants for eggs and first to third instar nymphs but decreased for fourth instars, fifth instars, and total nymphal life on CLas+ plants. The survival rates of eggs and first and second instar nymphs were significantly lower, while those of third to fifth instar nymphs were significantly higher on CLas+ plants. However, overall nymphal survival did not differ between CLas+ and CLas- plants. The pre-oviposition period of Asian citrus psyllid on CLas+ plants decreased, while oviposition period increased, resulting in significantly higher fecundity and population trend index compared with those on CLas- plants, illustrating an overall fitness benefit of CLas infection to Asian citrus psyllid. On the other hand, the longevity and resistance to starvation and lower temperatures of adults from CLas+ plants were significantly lower than those from CLas- plants suggesting that there may be a fitness cost for Asian citrus psyllid on CLas+ plants. The development time of eggs and nymphs and adult longevity were always shorter on C. reticulata regardless of infection status. This knowledge of host plant, pathogen, and vector interactions is useful for developing Asian citrus psyllid-HLB management programs.

Gut symbiont enhances insecticide resistance in a significant pest, the oriental fruit fly Bactrocera dorsalis (Hendel).

BACKGROUND: Symbiotic bacteria affect insect physiology and ecology. They may also mediate insecticide resistance within their hosts and thereby impact pest and vector control practices. Here, we document a novel mechanism of insecticide resistance in which a gut symbiont of the tephritid pest fruit fly Bactrocera dorsalis enhances resistance to the organophosphate insecticide trichlorphon. RESULTS: We demonstrated that the gut symbiont Citrobacter sp. (CF-BD) plays a key role in the degradation of trichlorphon. Based on a comparative genomics analysis with other Citrobacter species, phosphatase hydrolase genes were identified in CF-BD. These CF-BD genes had higher expression when trichlorphon was present. Bactrocera dorsalis inoculated with isolated CF-BD obtained higher trichlorphon resistance, while antibiotic-treated flies were less resistant confirming the key role of CF-BD in insecticide resistance. CONCLUSIONS: Our findings suggest that symbiont-mediated insecticide resistance can readily develop in B. dorsalis and may represent a more widely relevant insecticide resistance mechanism than previously recognized.

Analyses of Mitogenome Sequences Revealed that Asian Citrus Psyllids (Diaphorina citri) from California Were Related to Those from Florida.

Asian citrus psyllid (ACP, Diaphorina citri Kuwayama) transmits "Candidatus Liberibacter asiaticus" (CLas), an unculturable alpha-proteobacterium associated with citrus Huanglongbing (HLB). CLas has recently been found in California. Understanding ACP population diversity is necessary for HLB regulatory practices aimed at reducing CLas spread. In this study, two circular ACP mitogenome sequences from California (mt-CApsy, ~15,027 bp) and Florida (mt-FLpsy, ~15,012 bp), USA, were acquired. Each mitogenome contained 13 protein coding genes, 2 ribosomal RNA and 22 transfer RNA genes, and a control region varying in sizes. The Californian mt-CApsy was identical to the Floridian mt-FLpsy, but different from the mitogenome (mt-GDpsy) of Guangdong, China, in 50 single nucleotide polymorphisms (SNPs). Further analyses were performed on sequences in cox1 and trnAsn regions with 100 ACPs, SNPs in nad1-nad4-nad5 locus through PCR with 252 ACP samples. All results showed the presence of a Chinese ACP cluster (CAC) and an American ACP cluster (AAC). We proposed that ACP in California was likely not introduced from China based on our current ACP collection but somewhere in America. However, more studies with ACP samples from around the world are needed. ACP mitogenome sequence analyses will facilitate ACP population research.

The complete mitochondrial genome sequence of Diaphorina citri (Hemiptera: Psyllidae).

The first complete mitochondrial genome (mitogenome) sequence of Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae) from Guangzhou, China, is presented. The circular mitogenome is 14,996 bp in length with an A + T content of 74.5%, and contains 13 protein-coding genes (PCGs), 22 tRNA genes (tRNAs), two rRNA genes and an AT-rich control region. Synteny was identical to the ancestral insect mitogenomes. All PCGs start with 'ATN' codons. Eleven PCGs use the complete termination codons ('TAA' or 'TAG'), while the remaining two (cox2 and nad5) use a single 'T' as the stop codon. All tRNAs have the typical cloverleaf structure, except for trnTrp which lacks the variable arm and trnSer (AGN) which lacks the dihydrouridine arm. A phylogenetic tree, including 12 members of Sternorrhyncha, was constructed based on the 13 PCGs sequences. The resulting phylogenetic tree matched the established taxonomic scheme based on morphology.

The Complete Mitochondrial Genome Sequence of Bactericera cockerelli and Comparison with Three Other Psylloidea Species.

Potato psyllid (Bactericera cockerelli) is an important pest of potato, tomato and pepper. Not only could a toxin secreted by nymphs results in serious phytotoxemia in some host plants, but also over the past few years B. cockerelli was shown to transmit "Candidatus Liberibacter solanacearum", the putative bacterial pathogen of potato zebra chip (ZC) disease, to potato and tomato. ZC has caused devastating losses to potato production in the western U.S., Mexico, and elsewhere. New knowledge of the genetic diversity of the B. cockerelli is needed to develop improved strategies to manage pest populations. Mitochondrial genome (mitogenome) sequencing provides important knowledge about insect evolution and diversity in and among populations. This report provides the first complete B. cockerelli mitogenome sequence as determined by next generation sequencing technology (Illumina MiSeq). The circular B. cockerelli mitogenome had a size of 15,220 bp with 13 protein-coding gene (PCGs), 2 ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs), and a non-coding region of 975 bp. The overall gene order of the B. cockerelli mitogenome is identical to three other published Psylloidea mitogenomes: one species from the Triozidae, Paratrioza sinica; and two species from the Psyllidae, Cacopsylla coccinea and Pachypsylla venusta. This suggests all of these species share a common ancestral mitogenome. However, sequence analyses revealed differences between and among the insect families, in particular a unique region that can be folded into three stem-loop secondary structures present only within the B. cockerelli mitogenome. A phylogenetic tree based on the 13 PCGs matched an existing taxonomy scheme that was based on morphological characteristics. The available complete mitogenome sequence makes it accessible to all genes for future population diversity evaluation of B. cockerelli.

Density-dependent benefits in ant-hemipteran mutualism? The case of the ghost ant Tapinoma melanocephalum (Hymenoptera: Formicidae) and the invasive mealybug Phenacoccus solenopsis (Hemiptera: Pseudococcidae).

Although density-dependent benefits to hemipterans from ant tending have been measured many times, few studies have focused on integrated effects such as interactions between ant tending, natural enemy density, and hemipteran density. In this study, we tested whether the invasive mealybug Phenacoccus solenopsis is affected by tending by ghost ants (Tapinoma melanocephalum), the presence of parasitoids, mealybug density, parasitoid density and interactions among these factors. Our results showed that mealybug colony growth rate and percentage parasitism were significantly affected by ant tending, parasitoid presence, and initial mealybug density separately. However, there were no interactions among the independent factors. There were also no significant interactions between ant tending and parasitoid density on either mealybug colony growth rate or percentage parasitism. Mealybug colony growth rate showed a negative linear relationship with initial mealybug density but a positive linear relationship with the level of ant tending. These results suggest that benefits to mealybugs are density-independent and are affected by ant tending level.

Si-CSP9 regulates the integument and moulting process of larvae in the red imported fire ant, Solenopsis invicta.

Chemosensory proteins (CSPs) have been predicted to be involved in development; however, direct evidence for their involvement is lacking, and genetic basis is largely unknown. To determine the function of the chemosensory protein 9 (Si-CSP9) gene in Solenopsis invicta, we used RNA interference to silence Si-CSP9 in 3rd-instar larvae. The 3rd-instar larvae failed to shed their cuticle after being fed Si-CSP9-directed siRNA, and expression profiling of RNAi-treated and untreated control larvae showed that 375 genes were differentially expressed. Pathway enrichment analysis revealed that 4 pathways associated with larval development were significantly enriched. Blast analysis revealed that one fatty acid amide hydrolase (FAAH) gene was up-regulated and 4 fatty acid synthase (FAT) genes and one protein kinase DC2 gene (PKA) were down-regulated in the enriched pathways. Significantly higher expression of these genes was found in 4th-instar larvae, and Pearson correlation analysis of the expression patterns revealed significant relationships among Si-CSP9, PKA, FAAH, and FAT1-4. Moreover, we confirmed that expression levels of Si-CSP9, FAAH, and FAT1-4 were significantly reduced and that the development of 3rd-instar larvae was halted with PKA silencing. These results suggest that Si-CSP9 and PKA may be involved in the network that contributes to development of 3rd-instar larvae.

Effect of temperature and grain type on the long-term persistence and efficacy of s-methoprene in controlling Rhyzopertha dominica.

BACKGROUND: The efficacy of methoprene can vary with surface substrates, application methods and environmental conditions. The objectives of the present study were to examine the effects of temperature and grain type on the long-term persistence and efficacy of methoprene in controlling Rhyzopertha dominica from Guangzhou, China. RESULTS: Methoprene applied at 1 mg kg(-1) caused >90% suppression of F1 adult progeny of R. dominica for 150-270 days. Temperature and grain type both influenced the long-term persistence of methoprene. Overall multivariate analysis of variance showed that the order of progeny reduction at different temperatures (°C) was: 24 > 28 > 32 > 36; the order of the progeny reduction on the different grains was: paddy > wheat and maize. CONCLUSION: The results of our experiments show the maximum effect of methoprene for R. dominica control at 24 °C on paddy.

Association between changes in reproductive activity and D-glucose metabolism in the tephritid fruit fly, Bactrocera dorsalis (Hendel).

Reproduction is an important life process in insects; however, few studies have attempted to demonstrate the association between reproductive activity and energy metabolism. To address this problem, we focused on the reproductive changes in Bactrocera dorsalis males. We analyzed B. dorsalis male gene expression profiles during mating (DM), 3 h after mating (A3HM) and 12 h after mating (A12HM). Gene annotation and pathway analyses of differentially expressed genes show that galactose metabolism and the starch and sucrose metabolism pathway activities were significantly higher in A12HM group. Moreover, the maltase D gene was the most strongly up-regulated gene. The D-glucose levels were significantly higher in A12HM group. Maltase D expression level was significantly higher in males reared with sucrose. Body weights of the males reared with D-glucose and sucrose were significantly higher than those of the males reared with yeast extract. We observed more mated males from the groups fed sucrose and D-glucose than from those fed yeast extract. The D-glucose levels in individual males were highest at 18:00 h, when flies exhibit the most active mating behavior. This study shows that the maltase D gene and D-glucose are the critical gene and substrate, respectively, in male B. dorsalis mating process.

Solenopsis invicta (Hymenoptera: Formicidae), defend Phenacoccus solenopsis (Hemiptera: Pseudococcidae) against its natural enemies.

Mutualism is a common and important ecological phenomenon characterized by beneficial interaction between two species. Red imported fire ants, Solenopsis invicta Buren, tend honeydew-producing hemipteran insects and reduce the activity of these insects' enemies. Ant-hemipteran interactions frequently exert positive effects on the densities of hemipterans. We tested the hypothesis that ant tending can increase the densities of the mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae), and reduce the densities of the mealybug's predatory and parasitic enemies, the lady beetle, Menochilus sexmaculata Fabricius (Coleoptera: Coccinellidae), and the parasitoid wasp, Aenasius bambawalei Hayat (Hymenoptera: Encyrtidae). We found that more ants foraged on mealybug-infested hibiscus plants than on mealybug-free plants. The number of foraging ants on plants infested with high densities of mealybugs (62.5 ants per plant) was nearly six times that on mealybug-free plants (10.2 ants per plant). Experiment results showed that ant tending significantly increased the survival of mealybugs: if predatory and parasitic enemies were present, the survival of mealybugs tended by fire ants was higher than that in the absence of tending ants. Furthermore, this tending by fire ants significantly decreased the survival of lady beetle larvae. However, no apparent effect was observed on the survival of parasitoid.

Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues.

To accurately assess gene expression levels, it is essential to normalize real-time quantitative PCR (RT-qPCR) data with suitable internal reference genes. For the red imported fire ant, Solenopsis invicta, reliable reference genes to assess the transcript expression levels of the target genes have not been previously investigated. In this study, we examined the expression levels of five candidate reference genes (rpl18, ef1-beta, act, GAPDH, and tbp) in different developmental stages, castes and tissues of S. invicta. To evaluate the suitability of these genes as endogenous controls, three software-based approaches (geNorm, BestKeeper and NormFinder) and one web-based comprehensive tool (RefFinder) were used to analyze and rank the tested genes. Furthermore, the optimal number of reference gene(s) was determined by the pairwise variation value. Our data showed that two of the five candidate genes, rpl18 and ef1-beta, were the most suitable reference genes because they have the most stable expression among different developmental stages, castes and tissues in S. invicta. Although widely used as reference gene in other species, in S. invicta the act gene has high variation in expression and was consequently excluded as a reliable reference gene. The two validated reference genes, rpl18 and ef1-beta, can be widely used for quantification of target gene expression with RT-qPCR technology in S. invicta.

Fire ants protect mealybugs against their natural enemies by utilizing the leaf shelters constructed by the leaf roller Sylepta derogata.

The importance of mutualism is receiving more attention in community ecology. In this study, the fire ant Solenopsis invicta was found to take advantage of the shelters constructed by the leaf roller Sylepta derogata to protect mealybugs (Phenacoccus solenopsis) against their natural enemies. This protective effect of fire ant tending on the survival of mealybugs in shelters was observed when enemies and leaf rollers were simultaneously present. Specifically, fire ants moved the mealybugs inside the shelters produced by S. derogata on enemy-infested plants. Compared with that in plants without ants, the survival of mealybugs in shelters in the presence of natural enemies in plants with ants markedly improved. Both the protection of ants and the shelters provided by leaf rollers did not affect the survival of mealybugs in the absence of enemies in plants. Ants and leaf rollers significantly improved the survival of mealybugs in predator-infested plants, whereas no such improvement was observed in parasitoid-infested ones.

Does mutualism drive the invasion of two alien species? The case of Solenopsis invicta and Phenacoccus solenopsis.

Although mutualism between ants and honeydew-producing hemipterans has been extensively recognized in ecosystem biology, however few attempts to test the hypothesis that mutualism between two alien species leads to the facilitation of the invasion process. To address this problem, we focus on the conditional mutualism between S. invicta and P. solenopsis by field investigations and indoor experiments. In the laboratory, ant colony growth increased significantly when ants had access to P. solenopsis and animal-based food. Honeydew produced by P. solenopsis also improved the survival of ant workers. In the field, colony density of P. solenopsis was significantly greater on plots with ants than on plots without ants. The number of mealybug mummies on plants without fire ants was almost three times that of plants with fire ants, indicating a strong effect of fire ants on mealybug survival. In addition, the presence of S. invicta successfully contributed to the spread of P. solenopsis. The quantity of honeydew consumption by S. invicta was significantly greater than that of a presumptive native ant, Tapinoma melanocephalum. When compared with the case without ant tending, mealybugs tended by ants matured earlier and their lifespan and reproduction increased. T. melanocephalum workers arrived at honeydew more quickly than S. invicta workers, while the number of foraging S. invicta workers on plants steadily increased, eventually exceeding that number of T. melanocephalum foragers. Overall, these results suggest that the conditional mutualism between S. invicta and P. solenopsis facilitates population growth and fitness of both species. S. invicta tends to acquire much more honeydew and drive away native ants, promoting their predominance. These results suggest that the higher foraging tempo of S. invicta may provide more effective protection of P. solenopsis than native ants. Thus mutualism between these two alien species may facilitate the invasion success of both species.

Insecticide resistance of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), in mainland China.

BACKGROUND: In order to investigate the extent of resistance of oriental fruit fly, Bactrocera dorsalis (Hendel), which is a widespread pest throughout tropical, subtropical and temperate fruit crops, 25 populations of this insect were collected from 13 sites in mainland China in 2007 and 2008. In addition, resistant strains were established that showed increasing development of resistance. RESULTS: Compared with the susceptible strain, one population of B. dorsalis expressed high resistance (RR = 70.4-fold), 16 populations expressed medium resistance (11.5-fold < RR < 25.8-fold) and eight populations had low resistance or remained susceptible (1.4-fold < RR < 8.9-fold) to trichlorphon. As regards ß-cypermethrin, one population showed high resistance (RR = 44.0-fold), nine populations expressed medium resistance (12.2-fold < RR < 28.4-fold), 14 populations expressed low or minor resistance (3.0-fold < RR < 9.7-fold) and one population remained susceptible (RR = 1.1-fold). As regards avermectin, five populations had developed high resistance (44.3-fold < RR < 104-fold), seven populations expressed medium resistance (11.4-fold < RR < 38.6-fold) and three populations expressed low or minor resistance (3.5-fold < RR < 5.6-fold). A continuous resistance selection in the laboratory strain showed that the resistance ratios to trichlorphon, ß-cypermethrin and avermectin were 71.6-fold, 333-fold and 70.4-fold respectively. CONCLUSION: The data provided a comprehensive survey of insecticide resistance in Bactrocera dorsalis in mainland China. All results suggested that early resistance management programmes should be established for restoring the efficacy of pesticide-based control measures.

[Effects of volatiles from chestnut on host preference of adult Conogethes punctiferalis (Lepidoptera: Pyralidae)].

Through field survey and the tests of behavioral response, EAG response, and multiple-choice oviposition, this paper studied the effects of volatiles from Nongda No. 1 chestnut (NC) and Heyuan oil chestnut (HC) on the host-selection behavior of adult Conogethes punctiferalis. The field survey in 2004-2008 showed that the moth-eaten rate of NC fruits by adult C. punctiferalis was 16.1%-25.3%, while that of HC fruits was less than 5%. The volatiles from NC fruits and leaves were more attractive to female than to male moths, and the fruit volatiles were more attractive than leaf volatiles. However, the volatiles from HC fruits and leaves were not attractive to both female and male moths. The EAG response showed that female moths had significantly higher response to NC fruit volatiles than to HC's, but male moths had no significant difference in this response. For both NC and HC, the EAG responses of female and male moths to fruit volatiles were higher than those to leaf volatiles. The number of eggs laid by female moths was much greater on NC fruits than on NC leaves and on HC fruits and leaves, but had no significant differences on the latter three.

[Population life parameters and relative fitness of alpharmethrin-resistant Bactrocera dorsalis strain].

Based on the observation of the development and reproduction of experimental populations of alpharmethrin-resistant and susceptible Bactrocera dorsalis strains at (28 +/- 2) degrees C, the life tables of the populations were established, and the differences in the relative ecological fitness of the two strains was analyzed. The results showed that comparing with alpharmethrin-susceptible B. dorsalis strain, the resistant one had longer durations of development stages, average generation, and previposition, its fecundity was not stable and lower, and the hatch of eggs and the emergence of adults were depressed. The fecundity of the susceptible and resistant strains was 324.0 and 240.7, respectively. Based on the life tables, the population trend index (I) of resistant strain was 79.10, while that of susceptible strain was 116.97. The net reproduction rate (R0) and the intrinsic rate of increase (rm) of resistant strain were 1414.00 and 7.9463, while those of susceptible strain were 2184.00 and 8.3809, respectively. The relation fitness of resistant strain was only 0.6474. All of the results revealed that the resistant strain was at an obvious inferior position of reproduction and development.

[Foraging intensity of ants in Solenopsis invicta Buren (Hymenoptera: Formiciddae) invaded and un-invaded habitats].

By the methods of bait (honey, peanut oil, sausage, and mealworm larvae) trap, this paper studied the searching time, recruitment time, and recruitment number of ants in 3 typical habitats invaded and un-invaded by red imported fire ant (Solenopsis invicta) in South China, and analyzed the effects of S. invicta invasion on the foraging intensity of native ants. In S. invicta invaded lawn, the searching time of ants for mealworm larvae and peanut oil was significantly shorter, compared with that in S. invicta un-invaded area. Less difference was observed in the searching time for the 4 baits between S. invicta invaded and un-invaded wasteland, but the recruitment time for peanut oil was significantly longer in invaded than in un-invaded area. The searching time and recruitment time of the ants for the 4 baits had less difference between the invaded and un-invaded litchi orchard. 30 min after setting bait traps, the recruitment number of S. invicta workers on peanut oil, mealworm larvae, and sausage in invaded lawn, and on peanut oil in invaded wasteland was larger than that of native ants, but no significant difference was found in the recruitment number of S. invicta workers and native ants on the baits in invaded litchi orchard.

[Relationships between mound size and captured ergate amount of Solenopsis invicta].

By the method of bait trapping, this paper studied the relationships between mound size (mound volume and mound surface base area) and captured ergate amount of Solenopsis invicta in wasteland and greenbelt. In wasteland, with the increase of mound size, the amount of captured S. invicta ergates increased rapidly at first, slowed down then, and kept stable at last. When the mound volume and surface base area were 11634 cm3 and 1308 cm2 respectively, the captured ergate amount reached the maximum, being 291 individuals per trap. Similar patterns were observed in greenbelt. The increase of captured S. invicta ergate amount slowed down rapidly when the mound volume was larger than 18089 cm3, and reached the maximum (232 individual per trap) when the mound volume was 25974 cm3. The Weibull equation could better describe the relationships between the mound volume and mound surface base area and the captured ergate amount of S. invicta.