Gut microbiota variation across generations regarding the diet and life stage in Harmonia axyridis (Coleoptera: Coccinellidae).

We attempt to determine the effect of the dietary switch from a native to non-native prey on the gut microbiota in the predaceous ladybird Harmonia axyridis larvae and adults and examine how the dietary effect may vary across generations. We fed H. axyridis with different diets, native aphid Megoura japonica (Matsumura) versus non-native mealybug Phenacoccus solenopsis (Tinsley), for 5 generations and sequenced microbes in the gut of the 3rd instar larvae and adults of the 1st, 3rd, and 5th generations. In addition, we identified microbes in M. japonica and P. solenopsis. The 2 prey species differed in microbial community as measured by abundances of prevalent microbial genera and diversity. In H. axyridis, abundances of some prevalent microbial genera differed between the 2 diets in the 1st and 3rd generations, but the difference disappeared in the 5th generation; this tendency is more obvious in adults than in larvae. Overall, gut microbial assemblages became gradually cohesive over generations. Microbial diversity differed between diets in the 1st and 3rd generations but became similar in the 5th generation. Major prevalent gut microbial genera are predicted to be associated with metabolic functions of H. axyridis and associated genera are more abundant for consuming the mealybug than the aphid. Our findings from this study suggest that the gut microbiota in H. axyridis is flexible in response to the dietary switch, but tends toward homogeneity in microbial composition over generations.

Red imported fire ants (Hymenoptera: Formicidae) cover inaccessible surfaces with particles to facilitate food search and transportation.

Eusocial insects have evolved diverse particle-use behaviors. A previous study reported that red imported fire ants, Solenopsis invicta Buren, deposited soil particles on substances treated with essential balm, a fire ant repellent. We hypothesized that S. invicta modifies inaccessible surfaces by covering them with soil particles to facilitate food search and transportation. Here, laboratory experiments were conducted to study the particle-covering behavior of S. invicta in response to viscose surfaces or surfaces treated with essential balm or liquid paraffin in the presence of real food (sausage) or non-food objects (acrylic plates). S. invicta workers deposited significantly more soil particles on these three types of treated surfaces than on untreated surfaces. In addition, significantly more particles were relocated on viscose and paraffin-smeared surfaces in the presence of food than in the presence of non-food objects. The particle-covering behavior on viscose surfaces was also observed in the field. Interestingly, when no soil particles were available, ants searched and transported food on viscose surfaces only if the surfaces were artificially covered with sufficient quantities of soil particles but could not do so on viscose surfaces without soil particles or with insufficient quantities of soil particles. In addition, ants actively relocated particles to cover viscose surfaces if the transportation distance was within 200 mm, whereas significantly fewer particles were relocated at longer transportation distances (400 mm). Our study provides a novel example of particle use by fire ants during foraging.

Food Transport of Red Imported Fire Ants (Hymenoptera: Formicidae) on Vertical Surfaces.

Many ants can cooperatively transport large food items (either coordinated or uncoordinated during transportation), which can be rarely observed in other animals besides humans. Although these behaviors have been extensively investigated on horizontal surfaces, few studies dealt with food transport on vertical surfaces. The red imported fire ant, Solenopsis invicta Buren, is an invasive ant species that commonly forages on trees. Our studies showed that S. invicta used multiple strategies to transport food items on vertical surfaces (tree trunks). Small food items (1 × 1 × 1 mm sausage) were carried and transported by individual ants, and larger food items were either collectively and directly transported or cut collaboratively first and small particles were then transported individually or collectively. Competition and deadlocks were frequently observed during individual and collective transport respectively. During cutting, groups of ants tightly fixed the food on the tree trunks by holding the edges of the food item, while other ants cut the food into smaller particles. All food items and particles were moved downward. We investigated the effects of food placement (placed on a platform or fixed on tree trunk), food shape (cuboid or flattened), particle sizes (0.45-1, 1-2, 2-3, or 3-4 mm), and placement height (20, 80, or 150 cm) on the food transport on tree trunks. Our studies are the first to show how fire ants transport food on a vertical surface, and may provide insights into the development of novel fire ant baiting systems that can be placed on tree trunks.

Food-burying behavior in red imported fire ants (Hymenoptera: Formicidae).

The food-burying behavior has been reported in many mammals and birds, but was rarely observed in invertebrates. The red imported fire ants, Solenopsis invicta Buren, is an invasive pest in many areas of the world that usually performing food-burying during the foraging processes. However, the impacted factors and measureable patterns of this behavior is largely unknown. In the present study, food-burying vs food-transport behaviors of Solenopsis invicta were observed under laboratory and field conditions. When starved (no food was provided for 37 days) in the laboratory, food (sausage) was consumed by large numbers of ants, and few burying behaviors were observed. However, when food was provided until satiation of the colonies, food-transport was suppressed and significantly more soil particles were relocated on the food and graph paper square (where the food was placed) when compared with these colonies exposed to starved conditions. Videotapes showed that soil particles (1.47 ± 0.09 mm2) were preferentially placed adjacent to (in contact with) the food items at the beginning; and after the edges were covered, ants transported significantly smaller soil particles (1.13 ± 0.06 mm2) to cover the food. Meanwhile, larger particles (1.96 ± 0.08 mm2) were pulled/dragged around (but not in contact with) the food. Interestingly, only a small number of ants, mainly the small workers, were involved in food-burying, and the ants tended to repeatedly transport soil particles. A total of 12 patterns of particle transport were identified, and soil particles were most frequently picked from the foraging arena and subsequently placed adjacent to the food. In the field, almost all released food was actively transported by Solenopsis invicta workers, and no burying behavior was observed. Our results show that the food-burying behavior of Solenopsis invicta may be associated with the suppressed foraging activity, and the burying task may be carried out by certain groups of workers.

Effect of Polyacrylamide/Attapulgite Composite on Foraging Behaviors of Formosan Subterranean Termites (Blattodea: Rhinotermitidae).

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.

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

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 attention has been paid on how environmental factors affect these insects when they pupate within the soil. The tea looper, Ectropis grisescens Warren (Lepidoptera: Geometridae), is a severe pest of tea plants and has caused huge economic losses in South China. The protocols described here aim to investigate, through multiple-choice bioassays, whether mature last-instar E. grisescens larvae can discriminate soil variables such as the substrate type and moisture content, and determine, through no-choice bioassays, the impact of the substrate type and moisture content on pupation behaviors and the emergence success of E. grisescens. The results would enhance the understanding of the pupation ecology of E. grisescens and may bring insights into soil-management tactics for suppressing E. grisescens populations. In addition, these bioassays can be modified to study the influences of various factors on the pupation behaviors and survivorship of soil-pupating pests.

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

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.

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

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.

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

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.

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

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.