Ikkei Shikano

Evaluation of the toxicity of 17 essential oils against Choristoneura rosaceana (Lepidoptera: Tortricidae) and Trichoplusia ni (Lepidoptera: Noctuidae)†

BACKGROUND The obliquebanded leafroller, Choristoneura rosaceana Harris, and the cabbage looper, Trichoplusia ni Hübner, are serious fruit and vegetable pests requiring multiple insecticide applications per year. To reduce non-target exposure to hazardous insecticides and to curb resistance development, alternative controls are required. Accordingly, a selection of 17 essential oils was screened against both lepidopteran pests, and the influence of azinphos-methyl resistance on essential oil toxicity to C. rosaceana was studied. RESULTS Of the 17 essential oils screened, patchouli oil (Pogostemon cablin Benth.) and thyme oil (Thymus vulgaris L.) were selected for further testing against C. rosaceana, whereas patchouli oil, garlic oil (Allium sativum L.) and lemongrass oil (Cymopogon nardus L.) were selected for further testing against T. ni. LC(50) and LD(50) values confirmed that patchouli oil was the most toxic to C. rosaceana larvae, with LC(50) = 2.8 µL mL(-1) and LD(50) = 8.0 µg insect(-1). Garlic oil was the most toxic oil to T. ni larvae with LC(50) = 3.3 µL mL(-1) and LD(50) = 22.7 µg insect(-1), followed by patchouli oil and lemongrass oil. Azinphos-methyl-resistant leafrollers were 1.5-fold more tolerant to patchouli oil and 2.0-fold more tolerant to thyme oil. CONCLUSIONS Based on these results, patchouli oil and other essential oils have sufficient efficacy to be considered as components of an essential oil-based insecticide that targets these lepidopteran pests.

Relationship between adult and larval host plant selection and larval performance in the generalist moth, Trichoplusia ni

Adult oviposition preferences are expected to correlate with host plant suitability for the development of their offspring. For most lepidopteran species, this is particularly important as the hatching neonate larvae of many species are relatively immobile. Thus, the site of oviposition chosen by a female adult can greatly influence the probability of survival for her offspring. In the present study, we investigated the oviposition preference of adult Trichoplusia ni moths for six plant species to determine whether they could accurately rank the suitability of the plants for larval development. We also compared oviposition preferences to neonate larval acceptance and preference to determine whether the adult host range matched that of larval diet breath. Our results indicate that in two-choice and no-choice tests adult T. ni were able to rank the plants accurately, with the exception of anise hyssop. However, when given a choice of all six plants together, they laid more eggs on a plant that was not suitable for larval survival. Larvae accepted and fed on all plants in no-choice tests, and accurately ranked them according to larval performance. We conclude that neonate larvae are better able than adults to rank plants according to larval performance, and that larval diet breadth is wider than the range of plants accepted by adults. We also provide a discussion of the reduced accuracy of adult oviposition preference with increased plant choices.

Dietary Mechanism behind the Costs Associated with Resistance to Bacillus thuringiensis in the Cabbage Looper, Trichoplusia ni

Beneficial alleles that spread rapidly as an adaptation to a new environment are often associated with costs that reduce the fitness of the population in the original environment. Several species of insect pests have evolved resistance to Bacillus thuringiensis (Bt) toxins in the field, jeopardizing its future use. This has most commonly occurred through the alteration of insect midgut binding sites specific for Bt toxins. While fitness costs related to Bt resistance alleles have often been recorded, the mechanisms behind them have remained obscure. We asked whether evolved resistance to Bt alters dietary nutrient intake, and if reduced efficiency of converting ingested nutrients to body growth are associated with fitness costs and variation in susceptibility to Bt. We fed the cabbage looper Trichoplusia ni artificial diets differing in levels of dietary imbalance in two major macronutrients, protein and digestible carbohydrate. By comparing a Bt-resistant T. ni strain with a susceptible strain we found that the mechanism behind reduced pupal weights and growth rates associated with Bt-resistance in T. ni was reduced consumption rather than impaired conversion of ingested nutrients to growth. In fact, Bt-resistant T. ni showed more efficient conversion of nutrients than the susceptible strain under certain dietary conditions. Although increasing levels of dietary protein prior to Bt challenge had a positive effect on larval survival, the LC50 of the resistant strain decreased when fed high levels of excess protein, whereas the LC50 of the susceptible strain continued to rise. Our study demonstrates that examining the nutritional basis of fitness costs may help elucidate the mechanisms underpinning them.

Genetic Resistance to Bacillus thuringiensis Alters Feeding Behaviour in the Cabbage Looper, Trichoplusia ni

Evolved resistance to xenobiotics and parasites is often associated with fitness costs when the selection pressure is absent. Resistance to the widely used microbial insecticide Bacillus thuringiensis (Bt) has evolved in several insect species through the modification of insect midgut binding sites for Bt toxins, and reports of costs associated with Bt resistance are common. Studies on the costs of Bt-resistance restrict the insect to a single artificial diet or host-plant. However, it is well documented that insects can self-select appropriate proportions of multiple nutritionally unbalanced foods to optimize life-history traits. Therefore, we examined whether Bt-resistant and susceptible cabbage loopers Trichoplusia ni differed in their nutrient intake and fitness costs when they were allowed to compose their own protein:carbohydrate diet. We found that Bt-resistant T. ni composed a higher ratio of protein to carbohydrate than susceptible T. ni. Bt-resistant males exhibited no fitness cost, while the fitness cost (reduced pupal weight) was present in resistant females. The absence of the fitness cost in resistant males was associated with increased carbohydrate consumption compared to females. We demonstrate a sex difference in a fitness cost and a new behavioural outcome associated with Bt resistance.

Impact of non-pathogenic bacteria on insect disease resistance: importance of ecological context

1. The aerial surface of plants is a habitat for large and diverse microbial communities; termed the phyllosphere. These microbes are unavoidably consumed by herbivores, and while the entomopathogens are well studied, the impact of non‐pathogenic bacteria on herbivore life history is less clear.

Topical application of a plant extract to different life stages of Trichoplusia ni fails to influence feeding or oviposition behaviour.

We have previously determined that larval feeding experience with a feeding/oviposition deterrent modified the feeding responses of larvae and oviposition responses of subsequent moths. These behavioural changes were attributed to learning, but the possibility of chemical legacy could not be ruled out. In the present study, we have topically applied a feeding/oviposition deterrent plant extract from Hoodia gordonii (Masson) Sweet ex Decne (Asclepiadaceae) to larvae, pupae, and adults of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) to determine whether the feeding response of larvae and oviposition response of subsequent female moths is similarly modified by chemicals applied to the external surface of the insect. Our results indicate that traces of the extract that may be present internally or externally on the larvae do not reduce the feeding deterrent response of larvae. Furthermore, traces of the extract in or on larvae, pupae, or adult moths did not alter oviposition choice of female moths, leading us to discount the role of experience through topical application in this study. The fact that feeding/oviposition choice was only influenced by prior feeding experience of the larvae and not by topical administration suggests that habituation via sensory stimulation through mouthpart chemosensilla is likely a central phenomenon. Continuous exposure of adult moths to the extract over a period of 7 days did not affect the oviposition response of the female moths, ruling out the role of adult experience on host‐plant selection in T. ni. To the best of our knowledge, this is the first study to examine the role of experience via topical application of chemicals onto all life stages of the insect except the egg. Chemical legacy may not be playing a role in influencing the oviposition choices of female T. ni moths.

Baculovirus-challenge and poor nutrition inflict within-generation fitness costs without triggering transgenerational immune priming.

Invertebrate hosts that survive pathogen challenge can produce offspring that are more resistant to the same pathogen via immune priming, thereby improving the fitness of their offspring in the same pathogen environment. Most evidence for immune priming comes from exposure to bacteria and there are limited data on other groups of pathogens. Poor parental nutrition has also been shown to result in the transgenerational transfer of pathogen resistance and increased immunocompetence. Here, we combine exposure to an insect DNA virus with a change in the parental diet to examine both parental costs and transgenerational immune priming. We challenged the cabbage looper, Trichoplusia ni, with a low dose of the baculovirus, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and altered dietary protein to carbohydrate ratio (p:c ratio) after virus exposure. Insects fed a low protein diet had lower haemolymph protein concentrations, and exhibited costs of smaller pupae and slower development, while survivors of virus challenge developed more slowly, irrespective of p:c ratio, and those that were virus-challenged and fed on a low protein diet showed a reduction in haemocyte density. In addition, AcMNPV-challenged parents laid fewer eggs earlier in egg laying although egg size was the same as for unchallenged parents. There was no evidence for increased resistance to AcMNPV (immune priming) or changes in haemocyte number (as proxy for constitutive cellular immunity) in the offspring either as a result of parental AcMNPV-challenge or low dietary p:c ratio. Therefore, although pathogen-challenge and nutritional changes can affect host development and reproduction, this does not necessarily translate into transgenerational immune priming. Our findings contrast with an earlier study on another type of baculovirus, a granulovirus, where immune priming was suggested. This indicates that transgenerational immune priming is not universal in invertebrates and is likely to depend on the host-pathogen system, or the level of pathogen exposure and the type of dietary manipulation. Identifying whether immune priming or transgenerational effects are relevant in field populations, remains a challenge.

Impact Of Environmental Variation On Host Performance Differs With Pathogen Identity: Implications For Host-Pathogen Interactions In A Changing Climate.

Specialist and generalist pathogens may exert different costs on their hosts; thereby altering the way hosts cope with environmental variation. We examined how pathogen-challenge alters the environmental conditions that maximize host performance by simultaneously varying temperature and nutrition (protein to carbohydrate ratio; P:C) after exposure to two baculoviruses; one that is specific to the cabbage looper, Trichoplusia ni (TnSNPV) and another that has a broad host range (AcMNPV). Virus-challenged larvae performed better on more protein-biased diets, primarily due to higher survival, whereas unchallenged larvae performed best on a balanced diet. The environmental conditions that maximized host performance differed with virus identity because TnSNPV-challenge inflicted fitness costs (reduced pupal weight and prolonged development) whereas AcMNPV-challenge did not. The performance of TnSNPV-challenged larvae rose with increasing P:C across all temperatures, whereas temperature modulated the optimal P:C in AcMNPV-challenged larvae (slightly protein-biased at 16 °C to increasingly higher P:C as temperature increased). Increasing temperature reduced pupal size, but only at more balanced P:C ratios, indicating that nutrition moderates the temperature-size rule. Our findings highlight the complex environmental interactions that can alter host performance after exposure to pathogens, which could impact the role of entomopathogens as regulators of insect populations in a changing climate.

Altered nutrient intake by baculovirus-challenged insects: Self-medication or compensatory feeding?

Infection by parasites can alter the feeding behaviour of hosts. Some animals seek out substances that can therapeutically clear infections (self-medication), some may seek out resources to recoup resources lost while fighting off infection (compensatory feeding) and others may be manipulated to ingest substances that benefit parasite fitness (parasite manipulation of host). Recent studies have indicated that pathogen-challenged insects can self-medicate by increasing their protein intake relative to carbohydrate, which is thought to act by boosting the insects immune response. However, increased protein intake could also be due to compensatory feeding or pathogen manipulation of the host, and a rigorous examination of all four of the testable predictions, which is necessary for verifying self-medication behaviour, has not been conducted. The therapeutic behaviour must (1) only be employed by infected individuals and (2) alleviate the potential fitness loss of the infected individual. (3) If an uninfected individual engages in the behaviour, they suffer a decrease in fitness, and lastly, (4) the parasite cannot benefit from the behaviour. In response to baculovirus-challenge (AcMNPV) at 24°C, the cabbage looper, Trichoplusia ni, increased proportional protein intake, by increasing protein intake rather than decreasing carbohydrate intake. Increased protein intake did not benefit virus fitness, but it also did not increase the probability of host survival. Increased proportional protein intake did not occur in response to TnSNPV-challenge at 24°C or in response to AcMNPV-challenge at a higher temperature (32°C), indicating that the virus-induced change in nutrient intake depends on virus identity and temperature. Since virus-challenged T. ni did not show the typical costs associated with infection, the altered nutrient intake is likely to be a compensatory response. Understanding the motivation behind pathogen-induced changes in feeding behaviour could have significant implications for determining its importance for species interactions at multiple trophic levels.

Dishabituating long-term memory for gustatory habituation in the cabbage looper, Trichoplusia ni.

The gustatory rejection response of the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae), habituates to antifeedant compounds, allowing for the consumption of deterrent yet nontoxic plant materials. In the present study, we demonstrate that habituation to an antifeedant compound (quinine) persists through the moult between larval instars. As an indirect test of whether the memory was protein synthesis-dependent, we tested whether disrupting protein synthesis would block memory reconsolidation after a reminder. The results indicated that disrupting protein synthesis in habituated larvae following a reminder treatment (reexposure to quinine) eliminated the memory for habituation and restored the antifeedant properties of the quinine. We then examined whether the learned memory could be dishabituated and whether this would disrupt or eliminate long-term memory. We show that 6 hr after exposure to a novel/noxious stimulus (a second antifeedant, xanthotoxin) habituated larvae showed a transient dishabituation-like effect in which the quinine deterred feeding again. However, this effect did not permanently eliminate the habituation produced by the extended exposure as larvae tested 72 hr after xanthotoxin exposure again showed a willingness to consume the quinine treated leaves, indicating that the earlier habituation was still present.