choice-no-choice-bioassays-to-study-pupation-preference-emergence

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of <em>Ectropis grisescens</em>

Many insects live above the ground as larvae and adults and as pupate below the ground. Compared to the above-ground stages of their life cycles, less ...

South China Agricultural University

The tea geometrid, Ectropis grisescens Warren, is one of the most severe defoliator insect pests in tea plantations, China. The use of insecticides, etc., is forbidden on organic tea plantations. No female-produced sex pheromones of E. grisescens had been previously identified. In the present study, female gland extracts were analyzed by gas chromatography coupled with electroantennographic detection (GC-EAD) and two-dimensional gas chromatography (GC×GC) using a time-of-flight mass spectrometric detector (TOFMS). Two components, (Z,Z,Z)-3,6,9-octadecatriene (Z3Z6Z9-18:Hy) and (Z,Z)-3,9-6,7-epoxyoctadecadiene (Z3Z9-6,7-epo-18:Hy), were identified from pheromone gland extracts, and their electrophysiological and behavioral activity evaluated. Under laboratory conditions, Z3Z9-6,7-epo-18:Hy elicited a stronger electrophysiological response than Z3Z6Z9-18:Hy. In the field, traps baited with Z3Z9-6,7-epo-18:Hy alone showed better results than traps baited with Z3Z6Z9-18:Hy, and the binary mixture of Z3Z9-6,7-epo-18:Hy and Z3Z6Z9-18:Hy in a ratio of 4:1 (approximate ratio of females emitting pheromone) caught more males than the single components or any other blends. This study showed that Z3Z6Z9-18:Hy and Z3Z9-6,7-epo-18:Hy are the sex pheromone components of E. grisescens and that they prove useful in developing alternative management tools for the pest. 

Analysis of Tea Geometrid (Ectropis grisescens) Pheromone Gland Extracts Using GC-EAD and GC×GC/TOFMS.

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.

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

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. 

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

Many previous studies have focused on the foraging behaviors and strategies of the red imported fire ants, Solenopsis invicta Buren on solid food or granular bait; little attention has been paid to how liquid sugar is fed upon. In the present study, behavioral responses of S. invicta to 25% sucrose water droplets were observed. Five foraging patterns were identified in S. invicta colonies under laboratory conditions: (i) no feeding, no sucrose water feeding was observed; (ii) surround feeding, ants surrounded and fed along the edge of the sucrose droplet; (iii) stacked feeding, ants stacked and fed along the edge of the sucrose droplet; (iv) droplet-break feeding, ants broke the liquid droplet and sucked sucrose water that spread on surface of the substance or soil particles previously transported by ants; and (v) cover feeding, whole surface of the sucrose droplet was covered by layers of feeding ants. This is the first time cover feeding in S. invicta has been reported, which obviously requires more ants compared to the other patterns. In addition, individual ants were tracked in videos under laboratory conditions, and behavioral repertoires that led to stacking, covering and droplet-breaking were identified and described. The field investigation showed that surround feeding was most frequently performed by S. invicta foragers; however, cover feeding was not observed under field conditions during this study. Both laboratory and field studies showed colony-level variations in sugar-water feeding.

How do groups of red imported fire ants (Hymenoptera: Formicidae) feed on a droplet of sugar water?

Insect sex pheromones play a crucial role in the mate finding and calling behavior of Lepidoptera pests. Currently, little is known about the chemical ecology of Diaphania angustalis Snellen (Lepidoptera: Crambidae), a severe and important defoliator attacking the medicinal plant, Alstonia scholaris. In the present study, the pheromone components of D. angustalis females were investigated using electrophysiological and behavioral methods. Distilled hexane extracts of female pheromone glands were analyzed through electroantennogram (EAG) and gas chromatography-electroantennogram detector (GC-EAD), and the active compounds were identified through gas chromatography-mass spectrometry (GC-MS). Production peak of female sex pheromone occurred on the third day of age at 5 h into the scotophase with the EAG test, and the hexane extracts were attractive to males in the wind tunnel test. GC-EAD analysis of virgin males to gland extracts that were subsequently evaluated showed two active compounds, (E,E)-10,12-hexadecadienal (E10E12-16:Ald) and (E,E)-10,12-hexadecadien-1-ol (E10E12-16:OH), based on comparison of retention time and mass spectrum, with suitable synthetic compounds. Under laboratory conditions, the blend of E10E12-16:Ald and E10E12-16:OH in a ratio of 9:1 elicited a stronger EAG response than other treatments or a single component. In the field, more male moths were captured by traps baited with the mixture of E10E2-16:Ald and E10E2-16:OH in a ratio of 9:1, whereas a mixture of 8:1 and 10:1 also caught males. Accordingly, E10E2-16:Ald and E10E2-16:OH were regarded as the major sex pheromone components in D. angustalis females.

Production, identification, and field evaluation of sex pheromone from calling females in Diaphania angustalis (Lepidoptera: Crambidae).

Ectropis grisescens Warren (Lepidoptera: Geometridae) is one of the most severe pests of tea plants in China. This species commonly pupates in soil; however, little is known about its pupation ecology. In the present study, choice and no-choice tests were conducted to investigate the pupation behaviors and emergence success of E. grisescens in response to different substrates (sand, sandy loam 1, sandy loam 2, and silt loam) and moisture contents (5, 20, 35, 50, 65, and 80%). Moisture-choice bioassays showed that significantly more E. grisescens individuals pupated in or on soil (sandy loam 1 and 2 and silt loam) that was at the intermediate moisture levels, whereas 5%- and 35%-moisture sand was significantly more preferred over 80%-moisture sand for pupating. Substrate-choice bioassays showed that sand was most preferred by E. grisescens individuals at 20%- and 80%-moisture levels, but no preference was detected among the four substrates at 50%-moisture content. No-choice tests showed that the percentage of burrowed E. grisescens individuals and pupation depth were significantly lower when soil was dry (20% moisture) or wet (80% moisture). In addition, 20%-moisture sandy loam 2 and silt loam significantly decreased the body water content of pupae and emergence success of adults compared to 50%-moisture content. However, each measurement (percentage of burrowed individuals, pupation depth, body water content, or emergence success) was similar when compared among different moisture levels of sand. Interestingly, pupae buried with 80%-moisture soil exhibited significantly lower emergence success than that were unburied.

Pupation Behaviors and Emergence Successes of Ectropis grisescens (Lepidoptera: Geometridae) in Response to Different Substrate Types and Moisture Contents.

The pine sawyer beetle, Monochamus alternatus Hope (Coleoptera: Cerambycidae) is a serious insect pest of pine trees by vectoring the pine wood nematode, Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle (Nematoda: Aphelenchoididae). Field studies were carried out during 2013-2015 in Guangdong (China) to evaluate the effectiveness of commercial attractants, effect of trap placement for monitoring M. alternatus, and temporal patterns of trap catch. Four commercial attractants, three trap placements (0, 1.5, and 3 m above ground) and different trapping distances (50, 200, 400, 600, and 900 m) from forest edge were evaluated for monitoring M. alternatus. Traps baited with a mixture of monochamol and plant volatiles captured significantly more beetles than traps baited with monochamol alone or plant volatiles alone. Traps set up at 1.5 m above the ground captured significantly more M. alternatus than those at 0 m and 3 m height. Based on 2,687 beetles trapped from two locations in 2013 and 2014, the female:male ratio was 2.9-4.1:1. The beetles' natural dispersal distance was approximately 100 m based on traps set at different distances from edge of the forest. Continuous monitoring over 38-wk period indicates the peak of adult M. alternatus emergence was between May and June within a year.

Field Evaluation of Commercial Attractants and Trap Placement for Monitoring Pine Sawyer Beetle, Monochamus alternatus (Coleoptera: Cerambycidae) in Guangdong, China.

Previous studies showed that many wood-rotting fungi were attractive to termites; however, little attention has been paid to the relationship between termites and soil fungus. In the present study, different designs of two-choice tests were conducted to investigate the behaviors of two subterranean termites, Coptotermes formosanus Shiraki (wood-feeding lower termites) and Odontotermes formosanus (Shiraki) (fungus-growing higher termites), in response to soil (or sand) treated with the commercial conidial formulations of Trichoderma harzianum Rifai (BioWorks) and Trichoderma viride Pers. ex Fries (Shuiguxin). The short-term (1 d) choice tests showed no significant difference in termite aggregation (C. formosanus and O. formosanus) between treated and untreated soil, regardless of Trichoderma species and concentrations. However, in the long-term choice tests, C. formosanus consumed significantly more wood in the chambers containing soil treated with the conidial formulation of T. viride (1 × 108 conidia/g) than that containing untreated soil. The tunneling choice tests showed that sand treated with T. viride (1 × 106 or 1 × 108 conidia/g) or T. harzianum (1 × 106 conidia/g) significantly increased the tunneling activities of C. formosanus. However, sand treated with T. viride (1 × 106 or 1 × 108 conidia/g) had a repellent effect on O. formosanus. Our study showed that the two subterranean termites behaved differently when responding to the conidial formulations of Trichoderma.

Does Soil Treated with Conidial Formulations of Trichoderma spp. Attract or Repel Subterranean Termites?

The handling of sewage sludge (SS) and urban plant litter (UPL) has become an important concern. Immobilizing heavy metals (HMs) is regarded as a necessary process for recycling SS in agriculture and forestry. Here, HM removal and HM phytotoxicity in SS during vermicomposting with different additive UPLs was investigated. The results show that vermicomposting with additive UPL significantly reduced the content of HMs, and increased organic carbon content and the proportion of macroaggregates in SS. This process also significantly immobilized HMs by mainly transforming extractable and reducible HMs into residual products. The litters of Dracontomelon duperreanum and Bauhinia purpurea increased oxidizable HMs in SS and the accumulation capacity of HMs of earthworms during vermicomposting. The Cd content in vermicomposts with the B. purpurea litter addition was decreased by 31% relative to the initial SS. Maize in vermicomposts with UPL additions, especially with B. purpurea litter, exhibited significan5tly higher seed germination rates, seedling biomass, root activity, and a lower accumulation of HMs than in SS compost without UPL additions. These results suggest that vermicomposting with additive UPL can alleviate the phytotoxicity of HMs in SS and provides a new method for simultaneously recycling SS and UPL.

Alleviation of heavy metal phytotoxicity in sewage sludge by vermicomposting with additive urban plant litter.

The escaping behavior of termites has been documented under laboratory conditions; however, no study has been conducted in a field setting due to the difficulty of observing natural behaviors inside wood or structures (e.g., nests, tunnels, etc.). The black-winged termite,  (Shiraki), is a subterranean macrotermitine species which builds extensive mud tubes on tree trunks. In the present study, 41 videos (totaling ∼2,700 min) were taken on 22 colonies/subcolonies of  after their mud tubes were partially damaged by hand. In general, termites consistently demonstrated three phases of escape, including initiation (wandering near the mud-tube breach), individual escaping (single termites moving downward), and massive, unidirectional escaping flows (groups of termites moving downward). Downward moving and repairing were the dominant behavioral activities of individuals and were significantly more frequent than upward moving, turning/backward moving, or wandering. Interestingly, termites in escaping flows moved significantly faster than escaping individuals. Repairing behavior was observed shortly after the disturbance, and new mud tubes were preferentially constructed from the bottom up. When predators (i.e., ants) were present, however, termites stopped moving and quickly sealed the mud-tube openings by capping the broken ends. Our study provides an interesting example that documents an animal (besides humans) simultaneously carrying out pathway repairs and emergency evacuation without congestion.

Escaping and repairing behaviors of the termite  (Blattodea: Termitidae) in response to disturbance.

Subterranean termites often transport soil into bait stations. In this study, we hypothesize that adding soil or clay material in the bait may affect preference and performance of termites. Choice and no-choice tests were conducted in the laboratory to investigate the aggregation and feeding preference, survivorship, wood consumption, and body water percentage of termites in response to food containers (here we simulated the bait station by placing a wood block into a bigger plastic box with termite-entering holes on the wall) with the void space filled with soil (sandy clay loam), clay material (sodium bentonite), or remained unfilled. Choice tests showed that under low-moisture conditions, food containers filled with clay attracted significantly more termites (Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae) and Reticulitermes guangzhouensis Ping (Blattodea: Rhinotermitidae)) compared to food containers filled with soil, or unfilled. Under medium-moisture conditions, however, the percentages of termites that aggregated in the food containers filled with soil or clay were similar, and both were significant higher than the percentages in unfilled ones. In no-choice tests, the highest survivorship and wood consumption in C. formosanus were recorded under medium-moisture conditions and when food containers were filled with clay, whereas the lowest survivorship and wood consumption were recorded under low-moisture conditions and when food containers were filled with soil. Interestingly, presence of clay increased the body water percentage of termites. Our study enhances the understanding of the foraging ecology of subterranean termites, and may contribute to the improvement of termite-baiting technologies.

Filling Voids in Subterranean Termite (Blattodea: Rhinotermitidae) Bait Stations With Soil or Clay Improves Preference and Performance.

Moisture conditions of food and soil are essential for the survival and foraging activities of subterranean termites. Polyacrylamide/attapulgite composite is a water-retaining agent that has been applied to increase moisture of agricultural soils. We hypothesize that polyacrylamide/attapulgite composite may increase the moisture of bait matrixes and soil surrounding baiting containers and therefore attract termites and increase their foraging activities. In the present study, aggregation and feeding preferences, survivorship, body water percentage, and wood consumption of Formosan subterranean termites, Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae), were investigated when responding to polyacrylamide/attapulgite composite that was buried within soil (substrate) or filled in the void volume of baiting containers. Two-choice tests showed that termites consumed significantly more wood when polyacrylamide/attapulgite composite was buried within dry soil (27%-moisture) than the controls (no polyacrylamide/attapulgite composite was provided). However, polyacrylamide/attapulgite composite buried within wet soil (54%-moisture) did not significantly affect foraging behaviors of termites. Multiple-choice tests showed no aggregation or feeding preference of termites in the baiting containers filled with polyacrylamide/attapulgite composite compared with the soil-filled or unfilled ones, whenever the substrate was dry or wet. No-choice tests showed that the presence of polyacrylamide/attapulgite composite (buried within soil or filled in baiting containers) significantly increased wood consumption and body water percentage of termites as well as wood moisture under dry-substrate conditions. Our studies suggest that burying polyacrylamide/attapulgite composite within dry soil may enhance foraging activities of termites, but filling the bait stations with polyacrylamide/attapulgite composite may not effectively attract termites.

Article	Year
Analysis of Tea Geometrid (Ectropis grisescens) Pheromone Gland Extracts Using GC-EAD and GC×GC/TOFMS. Journal of agricultural and food chemistry\| PubMed ID: 27040982	2016
Essential Balm: A Strong Repellent Against Foraging and Defending Red Imported Fire Ants (Hymenoptera: Formicidae). Journal of economic entomology\| PubMed ID: 27298425	2016
Impact of Ground-Applied Termiticides on the Above-Ground Foraging Behavior of the Formosan Subterranean Termite. Insects\| PubMed ID: 27571108	2016
How do groups of red imported fire ants (Hymenoptera: Formicidae) feed on a droplet of sugar water? Insect science\| PubMed ID: 28028926	2018
Production, identification, and field evaluation of sex pheromone from calling females in Diaphania angustalis (Lepidoptera: Crambidae). Environmental science and pollution research international\| PubMed ID: 28900809	2017
Pupation Behaviors and Emergence Successes of Ectropis grisescens (Lepidoptera: Geometridae) in Response to Different Substrate Types and Moisture Contents. Environmental entomology\| PubMed ID: 29069472	2017
Field Evaluation of Commercial Attractants and Trap Placement for Monitoring Pine Sawyer Beetle, Monochamus alternatus (Coleoptera: Cerambycidae) in Guangdong, China. Journal of economic entomology\| PubMed ID: 29186543	2018
Does Soil Treated with Conidial Formulations of Trichoderma spp. Attract or Repel Subterranean Termites? Journal of economic entomology\| PubMed ID: 29471445	2018
Alleviation of heavy metal phytotoxicity in sewage sludge by vermicomposting with additive urban plant litter. The Science of the total environment\| PubMed ID: 29573693	2018
Escaping and repairing behaviors of the termite (Blattodea: Termitidae) in response to disturbance. PeerJ\| PubMed ID: 29576978	2018
Filling Voids in Subterranean Termite (Blattodea: Rhinotermitidae) Bait Stations With Soil or Clay Improves Preference and Performance. Journal of economic entomology\| PubMed ID: 29939283	2018
Effect of Polyacrylamide/Attapulgite Composite on Foraging Behaviors of Formosan Subterranean Termites (Blattodea: Rhinotermitidae). Journal of economic entomology\| PubMed ID: 30346605	2018

Cai Wang

Biography

JoVE Journal Publications

Other Publications