Alternative Nesting Strategies of Polistine Wasps in a Subtropical Locale.

Phylogenetic studies suggest that historically all paper wasps (Vespidae: Polistinae) in North America have tropical origins, but some species have adapted to survive temperate conditions. Subtropical climates, which are intermediate between temperate and tropical, allow a unique opportunity to study ancestral traits which can be retained or lost within populations, and ultimately elucidate the process of social wasp evolution. We investigated the phenology of paper wasps at study sites in subtropical Baton Rouge, USA, through nest searching and monitoring of nest parameters throughout the warm season (March-October). Across the year, two periods of nest initiation occurred: from March-May (early season nests, i.e., before the summer solstice), and from July-September (late season nests, after the solstice). We observed 240 Polistes nests from six species, of which 50.8% were initiated in early season and 49.2% in late season. In contrast, Mischocyttarus mexicanus rarely built late season nests and had longer early season colony duration than Polistes bellicosus and P. dorsalis, which built more nests in the late season than early. Across all species, late season nests had significantly shorter colony duration (~87.6 days) than early season nests (~166 days), and only P. bellicosus had fewer adults at peak population in late season nests than in early season nests. Results indicate both a bivoltine colony cycle in Polistes of subtropical climates, as well as differences in nesting strategies between genera.

Residual Effects of Termiticides on Mortality of Formosan Subterranean Termite (Isoptera: Rhinotermitidae) on Substrates Subjected to Flooding.

Concerns on efficacies of termiticides used for soil treatment to prevent Formosan subterranean termite (Coptotermes formosanus Shiraki) infestations have prompted pest control companies to suggest that retreatments are necessary after flooding of homes. Therefore, to address concerns about the efficacy of termiticides after flooding, we designed a flooding simulation experiment in the laboratory. We used four formulated termiticides containing fipronil, imidacloprid, chlorantraniliprole, or bifenthrin as active ingredients (a.i.) and two colonies of field-collected C. formosanus for this study. Evaluations of each chemical at concentrations of 1, 10, and 25 ppm in both sand and soil were conducted in the laboratory by comparing termite mortalities in no-choice bioassays after exposure to flooded (for 1 wk) and unflooded substrates. Toxicity from bifenthrin and fipronil were not affected by flooding regardless of substrate type except at the lowest concentration tested. Toxicity from chlorantraniliprole was lower in flooded sand at 1 ppm but otherwise similar among flooding treatments. In flooded soil, toxicity from chlorantraniliprole was low at 1 ppm, but unexpectedly high in flooded conditions at 10 and 25 ppm. For all concentrations of imidacloprid-treated sand, mortality of C. formosanus was reduced after a flood. However, like chlorantraniliprole, 10 and 25 ppm of imidacloprid-treated soil in flooded conditions resulted in an increased toxicity on C. formosanus. Our study supports the idea that chemicals with a higher water solubility like imidacloprid may require a home to be retreated with less water-soluble termiticides or baits after a flood.

A Preliminary Investigation of the Interactions between the Brood Parasite Chalcoela iphitalis and Its Polistine Wasp Hosts.

The life history of Chalcoela iphitalis-a common brood parasite of social wasps-has been described in previous literature, but critical information regarding oviposition behavior and possible differential host parasitism remain cryptic. Here we report on infestation levels of this moth in field populations of paper wasps in Polistes and Mischocyttarus, as well as the oviposition behavior of the moths under a laboratory setting. We found evidence for differential parasitism between paper wasp genera in the field, with almost 50% nest infestation in P. bellicosus and no occurrences of moth infestation in M. mexicanus. Laboratory results revealed that oviposition occurs only at night and is stimulated by contact with the wasp nest or adult wasps. In this setting, eggs were laid largely on the substrate above or adjacent to the host nest, but not on the nest itself.

Impact of Ground-Applied Termiticides on the Above-Ground Foraging Behavior of the Formosan Subterranean Termite.

We conducted a laboratory study to determine the impact of ground-applied termiticides on the above-ground foraging behavior of Coptotermes formosanus. Two concentrations (1 and 10 ppm) each of three termiticides, viz. fipronil, imidacloprid and chlorantraniliprole, were tested. After one month post-treatment (fipronil 10 ppm was run for 12 days only and all other treatments were run for one month), fipronil had the lowest percentage of survival (3%-4%) at both concentrations. Termite survival ranged from 31% to 40% in the case of imidacloprid treatments and 10 ppm chlorantraniliprole. However, 1 ppm chlorantraniliprole did not cause significant mortality compared to the controls. Foraging on the bottom substrate was evident in all replicates for all chemicals initially. However, a portion of the foraging population avoided the ground treatment toxicants after several days of bottom foraging. Only the slower-acting non-repellents created this repellent barrier, causing avoidance behavior that was most likely due to dead termites and fungus buildup on the treated bottom substrate. Fipronil appeared more toxic and faster acting at the concentrations tested, thus limiting this repellent effect. Suggestions by the pest control industry in Louisiana that some non-repellents can create a repellent barrier stranding live termites above ground are supported by this laboratory study.

Essential Balm: A Strong Repellent Against Foraging and Defending Red Imported Fire Ants (Hymenoptera: Formicidae).

In the present study, the repellent effects of essential balm, a traditional medicine product in China, was tested against foraging and defending red imported fire ants, Solenopsis invicta Buren, under laboratory and field conditions. The laboratory study showed that both band- (width = 1 cm) and patch-smearing of essential balm at each concentration (0.5, 1, or 2 µl/cm(2)) significantly decreased the number of S. invicta foragers within the 6-h observation period. Moreover, band-smearing of 2 µl/cm(2) essential balm and patch-smearing of 0.5, 1, and 2 µl/cm(2) essential balm inhibited most S. invicta foraging activity at 3, 6, 6, and 24 h into the experiment, respectively. The field study showed that after a disturbance was created on the S. invicta mound, there were significantly less defending ants on the substance treated (patch-smeared) with 0.5, 1, and 2 µl/cm(2) essential balm than the controls, but the number of ants on the substance of these three concentrations was similar. Our study suggested that essential balm is a strong repellent against foraging and defending S. invicta and could be applied when temporary protection from S. invicta is needed.

Repellent Effect of Formic Acid Against the Red Imported Fire Ant (Hymenoptera: Formicidae): A Field Study.

Previous studies showed that the formic acid secreted by tawny crazy ants not only has fumigation toxicity to the red imported fire ant, Solenopsis invicta Buren (Chen et al. 2013), but also can detoxify fire ant venom (LeBrun et al. 2014). These lead us to a field study to determine if low concentrations of formic acid might be useful in repelling S. invicta. Filter paper discs treated with 1.3% or 5% formic acid (v: v) or distilled water (control) were placed on each of the 46 S. invicta mounds and a disturbance was created. For a minute or less, there were significantly more defending ants on the control discs than that on the paper discs treated with formic acid. After food was added and for the next 40 min, there were significantly more foraging ants on the control discs compared to the treated discs. At 50 min into the test, the number of foraging ants on the control and 1.3% formic acid-treated discs was similar, but both were significantly higher than that on the 5% formic acid-treated discs. In addition, the active foraging (≥10 ants stayed on or around the food) and burying behavior (soil particles were deposited around the food) continued to be inhibited by 5% formic acid. The potential application and ecological significant of this repellent effect is discussed.

Panic escape polyethism in worker and soldier Coptotermes formosanus (Isoptera: Rhinotermitidae).

Termites were the first animal to form societies. Two hundred million years of evolution provide for a multitude of innate social behaviors that can be experimentally dissected. These fine-tuned patterns of behavior are especially interesting when observing group decision making in the panic mode. In this study, we examined behavioral patterns of termites under panic conditions to gain insight into how an escape flow self-organizes. One hundred worker and 10 soldier Coptotermes formosanus were released into agar plates. After a disturbance was created most workers followed each other and ran along the wall of dishes, thus forming a unidirectional escape flow, whereas soldiers showed a significantly higher frequency of moving to the center of the arena or on periphery of the escape flow as compared to workers. Agonistic behavior was usually observed as soldiers moved to center or periphery. This is the first report on the behavioral repertoire of termites when panicked, with details on the behavioral polymorphism of workers and soldiers during an escape.

Point of No Return From Water Loss in Coptotermes formosanus (Isoptera: Rhinotermitidae).

Describing desiccation stages based on the physical appearance of termites has not been evaluated previously. Formosan subterranean termites were studied to determine the rate of water loss, singly and in groups, in the laboratory. The stages of water loss are described based on changes in physical appearance and percent total body water loss evaluated at 2- to 8-h time intervals up to 32 h. Workers in groups lost water slower than individual worker trials. Weight loss was linear over time for worker groups and individuals, as was individual soldier only trials. Water loss in individual workers was significantly faster than in soldiers. Three physical stages of desiccation are described for living workers: (I) curling of antennae, (II) on back but with assistance able to right themselves and walk, and (III) unable to get off back; and two stages for living soldiers (II and III). Recovery was determined from termites in a second trial by transferring stage I, II, and III individuals from open, dry Petri dishes to those with moist filter paper at 4, 6, 10, 12, 14, 16, 24, 26 and 28 h. After 12 h on moist filter paper, stage I workers had a 83% recovery rate, stage II had a 33%, and stage III had a 7% recovery. Soldiers had a 56% recovery at stage II and was similar to the recovery of workers at stage III. Most termites that reached stage III were destined to die.

A Kazal-Type Serine Protease Inhibitor from the Defense Gland Secretion of the Subterranean Termite Coptotermes formosanus Shiraki.

Coptotermes formosanus is an imported, subterranean termite species with the largest economic impact in the United States. The frontal glands of the soldier caste termites comprising one third of the body mass, contain a secretion expelled through a foramen in defense. The small molecule composition of the frontal gland secretion is well-characterized, but the proteins remain to be identified. Herein is reported the structure and function of one of several proteins found in the termite defense gland secretion. TFP4 is a 6.9 kDa, non-classical group 1 Kazal-type serine protease inhibitor with activity towards chymotrypsin and elastase, but not trypsin. The 3-dimensional solution structure of TFP4 was solved with nuclear magnetic resonance spectroscopy, and represents the first structure from the taxonomic family, Rhinotermitidae. Based on the structure of TFP4, the protease inhibitor active loop (Cys(8) to Cys(16)) was identified.

Water Transport by Coptotermes formosanus (Isoptera: Rhinotermitidae).

Subterranean termites are extremely vulnerable to desiccation, and high moisture makes their habitat and food favorable for survival and colony growth. Although there is a general perception that termites can manipulate moisture, documentation is surprisingly scanty with regard to how termites transport water and the factors that impact it. There has been no study of water transfer by Coptotermes formosanus Shiraki, a notoriously invasive termite in the southern United States. We conducted a study to determine if C. formosanus transfers water. Bioassays using arenas with a dry food source connected to a moist substrate by either a short tube (10 cm) or a long tube (100 cm) were conducted. Three moistened substrate types were tested to see how they impacted water transfer. In addition, workers and soldiers sampled from a moist sand substrate were dissected to determine water sac volumes for possible transfer of water to wood. The results indicated that some water transfer is achieved by the evacuation of water sacs. However, moist soil was also moved to increase humidity. When termites had use of moist silty clay, wood moisture gain increased significantly in both 10 and 100 cm tubes. As tube distance increased, moisture to the more distant food source decreased. Workers had the largest water sacs, though soldiers appear to contribute in water transfer via water sacs as well. Water transfer and its implications are discussed.

Lethal and sublethal effects of lufenuron on the Formosan subterranean termite (Isoptera: Rhinotermitidae).

A laboratory study was conducted to understand the effect of low concentrations of lufenuron on termite physiology and behavior. Survivorship, running speed, body water content, food consumption, tunneling, microbial infection, and two behavioral patterns (carcass-burying behavior and particle transport behavior) were compared among Formosan subterranean termites, Coptotermes formosanus Shiraki, fed lufenuron-treated (250, 500, or 1,000 ppm) or untreated (control) filter paper. In 30-32 d, all lufenuron treatments significantly reduced survivorship, running speed, consumption, and tunneling, but had no substantial effect on body water content. In addition, termites fed the three concentrations of lufenuron became infected by opportunistic pathogens. Carcass-burying and particle transport behaviors also were inhibited by lufenuron. Potential application of lufenuron at low concentrations for the control of C. formosanus is discussed.

Clay preference and particle transport behavior of Formosan subterranean termites (Isoptera: Rhinotermitidae): a laboratory study.

Although preference and utilization of clay have been studied in many higher termites, little attention has been paid to lower termites, especially subterranean termites. The Formosan subterranean termite, Coptotermes formosanus Shiraki, can modify its habitat by using clay to fill tree cavities. Here, the biological significance of clay on C. formosanus was investigated. Choice tests showed that significantly more termites aggregated in chambers where clay blocks were provided, regardless of colony group, observation period, or nutritional condition (fed or starved). No-choice tests showed that clay had no observable effect on survivorship, live or dry biomass, water content, and tunneling activity after 33-35 d. However, clay appeared to significantly decrease filter paper consumption (dry weight loss). Active particle (sand, paper, and clay) transport behavior was observed in both choice and no-choice tests. When present, clay was preferentially spread on the substrate, attached to the smooth surfaces of the containers, and used to line sand tunnels. Mechanisms and potential application of clay attraction are discussed.

myo-Inositol and Phytate Are Toxic to Formosan Subterranean Termites (Isoptera: Rhinotermitidae).

Several rare and common monosaccharides were screened for toxic effects on the Formosan subterranean termite, Coptotermes formosanus Shiraki, with the aim of identifying environmentally friendly termiticides. myo-Inositol and phytic acid, which are nontoxic to mammals, were identified as potential termite control compounds. Feeding bioassays with termite workers, where both compounds were supplied on filter paper in concentrations from 160.2 to 1,281.7 µg/mm(3), showed concentration-dependent toxicity within 2 wk. Interestingly myo-inositol was nontoxic when administered to termites in agar (40 mg/ml) in the absence of a cellulosic food source, an unexplained phenomenon. In addition, decreased populations of termite hindgut protozoa were observed upon feeding on myo-inositol but not phytate-spiked filter paper. Radiotracer feeding studies using myo-inositol-[2-(3)H] with worker termites showed no metabolism after ingestion over a 2-d feeding period, ruling out metabolites responsible for the selective toxicity.

Localized treatments using commercial dust and liquid formulations of fipronil against Coptotermes formosanus (Isoptera: Rhinotermitidae) in the laboratory.

Use of proper application methods and formulations of termiticides are important to reduce their negative impact to the environment. In this study, we conducted laboratory experiments to determine the effect of localized treatments with commercial dust and liquid formulations of fipronil against Formosan subterranean termites, Coptotermes formosanus Shiraki. The test arena consisted of a specially designed 16-chambered structure with a center chamber connected to 5 foraging chambers that themselves were connected to 10 additional foraging chambers. One peripheral chamber received a liquid or dust treatment and termites were released in the center chamber. Results showed that >91% of the termites were dead within the 9-d test period despite the localized treatment of only 1 foraging chamber. Termites that were still alive after 9 d were transferred to an untreated dish and held for 10 more days. The majority of those termites were dead and the rest were moribund on day 19. Regardless of the specific dish treated, both formulations of fipronil were found to be highly efficacious. Termites did not exhibit repellency to either formulation. Our results suggest that localized (or spot) treatment with either commercially available dust or liquid formulations of fipronil can be a viable option for control of a termite infestation where complete soil drenching is not desirable.

Lufenuron suppresses the resistance of Formosan subterranean termites (Isoptera: Rhinotermitidae) to entomopathogenic bacteria.

Pesticides can negatively affect insect immunity. Although studies show that Formosan subterranean termites, Coptotermes formosanus Shiraki, are resistant to microbial infections, the effects of pesticides on disease resistance is not well studied. In this study, C. formosanus previously fed lufenuron was exposed to each of the three entomopathogenic bacteria, Pseudomonas aeruginosa (Schroeter) Migula, Serratia marcescens Bizio, and Bacillus thuringiensis Berliner subsp. israelensis. We found that termite mortality was significantly higher and synergistic in the combination of lufenuron and P. aeruginosa compared with treatment of lufenuron or P. aeruginosa alone. Other bacteria and lufenuron combinations were not quite as effective. Interestingly, only in treatments without lufenuron did termites show carcass-burying behavior. The results indicate that lufenuron, a chitin synthesis inhibitor, can suppress Formosan subterranean termite resistance to P. aeruginosa. Possible suppression mechanisms are discussed.

Horizontal transfer of fipronil is enhanced with increased group size in Coptotermes formosanus (Isoptera: Rhinotermitidae).

Fipronil is a widely used insecticide for termite control. Although transfer of fipronil among termite cohorts has been investigated in previous studies, no study has yet focused on the influence of termite group size (density) on horizontal transfer. In this study, the mortality of donor and recipient Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) was compared among groups of 10, 25, and 50 workers. Most donor termites were dead within 20 h. There was a significantly higher mortality of recipient termites starting at 44 h when in bigger groups. LT50 and LT90 of recipient termites decreased with increase in group size, being significantly shorter in groups of 50 termites compared with groups of 10 termites. Moreover, the variance (within-group difference) of recipient mortality and lethal time estimations was lowest in the groups of 50 termites, indicating a more uniform horizontal transfer of fipronil by termites in bigger groups. Our findings suggest that group size has an influence on fipronil transfer among C. formosanus workers and should be considered as a variable of importance.

Evaluation of three bait materials and their food transfer efficiency in Formosan subterranean termites (Isoptera: Rhinotermitidae).

The consumption and food transfer efficiency of two commercially used termite bait materials, southern yellow pine wood and cardboard, and one potential bait material, maize (Zea mays L.) cob, were evaluated for use against the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), in the laboratory. In the no-choice test, the consumption of wood and cob was similar and significantly more than cardboard. Tunneling under the food sources was similar. In the two-choice test, the consumption was cob > wood, wood > cardboard, cob = cardboard, and tunneling under these choices was cob = wood, wood = cardboard, cob > cardboard. In the three-choice test, no significant difference was detected in consumption, but tunnels made under the cob were significantly more than wood and cardboard. Nile blue A was used to study food transfer of bait material among termite cohorts. Dyed cardboard, cob, or wood (0.1% Nile blue A) was provided to termites as food. Termites feeding on wood turned blue in significantly greater number at 6 h compared with cardboard and cob, but there was no significant difference after 12 h. Blue termites feeding on different bait materials were then collected and combined with undyed termites. When undyed (white) termites were placed with blue termites and food (wood block), termites turned blue in the same percentage regardless of original bait material fed on. However, when no food was provided (starvation group), the rate of white termites turning blue was dramatic; in dyed wood treatment, significantly more termites turned blue than that of cardboard, although neither were significantly different from cob. Our study is the first to show that, cob, an otherwise waste product of the food and biofuel industry, is as efficient as wood and cardboard as a termite bait matrix.

Toxicity and horizontal transfer of 0.5% fipronil dust against Formosan subterranean termites.

The toxicity and horizontal transfer of a new formulation of fipronil, 0.5% fipronil dust, was tested against Coptotermes formosanus Shiraki in the laboratory. The formulation was applied in three different ways: 1) Directly applied to termites (donors) and mixed with untreated termites (recipients) at three ratios, viz., 50 donors: 50 recipients, 20 donors: 80 recipients and 10 donors: 90 recipients. 2) Applied onto the surface of 3 mm thick sand or soil substrate in a petri dish and then topped with another 3 mm thick sand or soil layer whereupon termites were released. 3) Applied to the inner surface of a tube (either 5 cm or 15 cm long) that connected two foraging dishes, one containing dry sand and the other moist sand plus a wood block and termites were released into the dry sand dish. All donors and >93% of the recipients were dead by 42 h after treatment in the direct treatment experiment. Significant mortalities of both donors and recipients were observed at 5 h after treatment at all donor: recipient ratios. During this period, the mortality of the recipients (but not donors) at 10:90 was significantly lower than those at the other two ratios. All termites were dead at 65 h after exposure (HAE) on the sand treatment and at 190 HAE on soil treatment. More than 96% mortality was observed at 40 HAE on the sand treatment as compared with only 6% mortality onsoil treatment during the same time period. In the tube treatment experiment, > 97% mortality was observed at 90 h after release for both tube lengths as compared with < 3% mortality in controls. About half of the termites were dead by 15 h after release regardless of the tube length. Our results showed that 0.5% fipronil dust is nonrepellent and readily transferred from treated to nontreated termites.

Laboratory Study of the Influence of Substrate Type and Temperature on the Exploratory Tunneling by Formosan Subterranean Termite.

Using two-dimensional foraging arenas, laboratory tests were conducted to investigate the effect of soil type, soil moisture level and ambient temperature on the exploratory tunneling by Coptotermes formosanus Shiraki. In choice arenas consisting of two substrate types having two moisture levels each, and conducted at a constant temperature of 22 °C, a significantly greater proportion of termites aggregated in sand than in sandy loam. Similarly, the length of excavated tunnels was also increased in sand. In a given substrate, termite aggregation or tunnel length did not differ between 5% and 15% moisture levels. In no-choice tests, where three different substrates (sand, sandy loam and silt loam) were tested at two temperatures (22 °C and 28 °C), excavations were significantly greater in sand than either sandy loam or silt loam at 22 °C. Fewer primary tunnels were constructed in sandy loam than in sand and fewer branched tunnels than either in sand or silt loam. No significant difference in either tunnel length or number of primary or branched tunnels was found between these two temperatures.

Toxicity of seven termiticides on the formosan and eastern subterranean termites.

Using both topical application and substrate (sand) treatments the toxicities of seven new generation soil termiticides were evaluated to determine the LD50 and LC50 against two economically important subterranean termite species, eastern subterranean termite, Reticulitermes flavipes (Kollar), and Formosan subterranean termite, Coptotermes formosanus Shiraki. The lethal dose toxicity (LD50) rankings for R. flavipes from highest to lowest were: fipronil > bifenthrin > chlorantraniliprole > cyantraniliprole > imidacloprid > chlorfenapyr > indoxacarb; the rankings for C. formosanus were fipronil > imidacloprid > chlorantraniliprole > cyanthraniliprole> bifenthrin > chlorfenapyr > indoxacarb. The respective lethal concentration toxicity (LC50) rankings were fipronil > bifenthrin > chlorfenapyr > indoxacarb > cyantraniliprole > chlorantraniliprole > imidacloprid for R. flavipes; and fipronil > chlorfenapyr > bifenthrin >imidacloprid > cyantraniliprole > chlorantraniliprole > indoxacarb for C. formosanus. The study provides an opportunity to directly compare toxicity, action speed, and bioavailability among this group of newer generation soil termiticides.