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

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).

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 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.

Diaphania angustalis Snellen (Lepidoptera: Crambidae) has emerged as a very important pest of blackboard tree, Alstonia scholaris (L.) R. Br. (Apocynaceae), in China during the last two decades. Understanding its biology and behavior is crucial for designing effective and environmentally friendly pest management strategies. Under laboratory conditions [25-28°C, 12:12 (L:D) h, 75-80% RH], adults emerged during both light and dark hours with peak emergence occurring between the first and fifth hours of scotophase, and unmated males and females lived for a mean (±SE) 5.4 ± 0.4 and 5.3 ± 0.7 d, respectively. Female calling behavior was observed only during scotophase (peaking in the fifth hour) by 1- to 5-d-old females, and mating activities occurred 2-5 d after emergence, peaking on day 3. These behavioral characteristics could inform control programs targeting adults.

Calling and Mating Behavior of Diaphania angustalis (Lepidoptera: Crambidae).

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.

Zhaohui Sun

Guangdong Key Laboratory for Innovation Development and Utilization of Forest Plant Germplasm,
College of Forestry and Landscape Architecture,
Guangdong Key Laboratory for Innovation Development and Utilization of Forest Plant Germplasm, College of Forestry and Landscape Architecture

South China Agricultural University

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens