Protocol for rearing and using mosquitoes for flight path tracking and behavioral characterization in wind tunnel bioassays.

Mosquito behavioral assays are an important component in vector research and control tool development. Here, we present a protocol for rearing Anopheles mosquitoes, performing host-seeking behavioral bioassays, and collecting 3D flight tracks in a large wind tunnel. We describe steps for setting up host-seeking landing assays, both as a non-choice and as a dual-choice assay, and analyzing flight tracks. This protocol can be applied in the research of several behavioral traits, including nectar seeking, resting, mating, and oviposition behavior. For complete details on the use and execution of this protocol, please refer to Carnaghi et al.1.

Mosquito odour-baited mass trapping reduced malaria transmission intensity: a result from a controlled before-and-after intervention study.

BACKGROUND: Conventional vector control strategies have significantly reduced the malaria burden. The sustainability of these methods is currently challenged. Odour-based traps are emerging technologies that can complement the existing tools. Implementation of odour-based traps for mass trapping is limited due to the restricted range of vectors caught with available carbon dioxide-dependent lures, and the lack of comprehensive field studies. The objective of this study was to assess the impact of odour-mediated mass trapping targeting outdoor vectors, using a synthetic cattle urine lure that attracts a wide range of vector species in a variety of physiological states, on malaria prevalence and entomological parameters to determine malaria transmission intensities. METHODS: A controlled before-and-after study was conducted in two rural communities in southern Ethiopia. Baseline monthly entomological and seasonal cross-sectional malaria prevalence surveys were conducted in both communities for a year. Then, mass trapping of mosquitoes was conducted in one of the villages, while the monthly entomological surveillance and seasonal malaria prevalence surveys continued in both villages. Generalised linear mixed models were constructed and tested to determine which factors were significantly affected by the intervention. RESULTS: Mass trapping contributed to the reduction of the population of the principal malaria vector, Anopheles arabiensis, and the associated entomological indicators, the human bite rate (HBR) and the entomological inoculation rate (EIR), in the intervention village compared to the control village. The intervention village had an average HBR by An. arabiensis of 3.0 (95% CI 1.4-4.6) during the peak malaria transmission season, compared to 10.5 (95% CI - 0.5-21.5; P < 0.0001) in the control village. The intervention village (mean 0.02, 95% CI - 0.05-0.4.8) had a daily EIR eight times lower than the control village (mean 0.17, 95% CI), which likely contributed to the reduced malaria prevalence in the intervention community following its introduction by ca. 60% (95% CI 55-63). CONCLUSIONS: The combined use of odour-based mass trapping and conventional control strategies coincided with a reduction of human-vector contact and malaria prevalence, providing support for odour-baited technologies as a viable option for next-generation vector control tools. Further cluster-randomised control studies are recommended in different eco-epidemiological settings with varying malaria transmission intensities.

Visual and thermal stimuli modulate mosquito-host contact with implications for improving malaria vector control tools.

Malaria prevention relies on mosquito control interventions that use insecticides and exploit mosquito behavior. The rise of insecticide resistance and changing transmission dynamics urgently demand vector control innovation. To identify behavioral traits that could be incorporated into such tools, we investigated the flight and landing response of Anopheles coluzzii to human-like host cues. We show that landing rate is directly proportional to the surface area of thermal stimulus, whereas close-range orientation is modulated by both thermal and visual inputs. We modeled anopheline eye optics to theorize the distance at which visual targets can be detected under a range of conditions, and experimentally established mosquito preference for landing on larger targets, although landing density is greater on small targets. Target orientation does not affect landing rate; however, vertical targets can be resolved at greater distance than horizontal targets of the same size. Mosquito traps for vector control could be significantly enhanced by incorporating these features.

Aedes aegypti oviposition-sites choice under semi-field conditions.

Vector control is still the recommended approach to avoid arbovirus outbreaks. Herein, we investigate oviposition preferences of Aedes aegypti (Diptera: Culicidae) females under a semi-field structure Rio de Janeiro, Brazil. For that, in Experiment 1, we used two settings: 'Single items', which included as containers drain, beer bottle, bucket, car tyre, water tank, and a potted Peace Lily (Spathiphyllum wallisii) in a saucer with water, or 'Multiple containers', as an urban simulation, in which one drain, two additional beer bottles, and an extra plant pot saucer were added. Experiment 2 (sensory cues) used five variations of potted plant, each one varying in the range of sensory cues known to attract gravid females to oviposition containers. Our results indicate that gravid Ae. aegypti prefer to oviposit close to the ground and in open water containers with organic compounds from plant watering. Domestic large artificial containers containing tap water received significantly fewer eggs, except for the car tyre, which exhibited as many eggs as the potted plant. We also show that visual (potted plant shape) and olfactory clues (odour of the plant or from water containing organic matter) were equally attractive separately as were these stimuli together.

The influence of manure-based organic fertilisers on the oviposition behaviour of Anopheles arabiensis.

The rice agroecosystem provides suitable breeding habitat for many malaria vector species, and  rice-adjacent communities are consequently exposed to a greater malaria transmission risk than non-rice-associated communities. As part of efforts to expand rice production in Africa, sustainable and climate-adapted practices such as the System of Rice Intensification (SRI) are being promoted. SRI encourages the use of organic fertilisers (OFs) such as cow and chicken dung, as opposed to inorganic industrially produced fertilisers, due to their lower resource cost, apparent benefit to the rice agroecosystem and as a means to reduce the greenhouse gas emissions associated with the production of industrial fertilisers. However, the impact of OFs on mosquito fauna is not well documented and may have knock-on consequences on malaria transmission risk. Here, we demonstrate, using dual choice egg count assays, that both cow and chicken dung modulate the oviposition behaviour of Anopheles arabiensis, a major malaria vector in Sub-Saharan Africa. A significantly reduced proportion of eggs were laid in water treated with either cow or chicken dung compared to untreated water, with higher dung concentrations resulting in further reduced proportions. When presented in competition, significantly fewer eggs were laid in water treated with chicken dung than with cow dung. Moreover, there was no evidence of egg retention in any experiment, including in no-choice experiments where only dung-containing dishes were available. These results suggest both cow and chicken dung may act as oviposition deterrents to malaria vector species and that the application of manure-based OFs in rice agriculture may modulate the oviposition behaviour of An. gambiae s.l. within agroecosystems. Quantification of the ammonia present in dung-infused water showed higher concentrations were present in the chicken dung infusion, which may be one contributing factor to the difference in observed deterrence between the two dung types. Deterrence of mosquito oviposition in OF-treated farms may potentially affect the overall production of malaria vectors within rice fields and their contribution to local malaria transmission.

Case closed - Wrappings and encasement delays and reduces fly presence on body parts.

Homicide, particularly where a body has been concealed, is uniquely challenging for investigators to estimate the time of occurrence due to the methods employed by perpetrators to hide the body or its constituent parts from detection. The regularity of necrophagous insect lifecycles to determine minimum post-mortem interval (minPMI) is widely employed but remains an unreliable technique if used without a clear understanding of the factors that affect insect access and oviposition behaviour to concealed remains. The purpose of this study was to investigate the effect of wrapping body parts on fly colonisation and implications for minPMI calculations. Field studies were carried out using four treatments of pork (as surrogate body parts), in five replicates, one unwrapped, the other three wrapped in either a black plastic sack, a small-zipped wash bag (to simulate a suitcase), or a plastic sack further placed in a wash bag. Over a 48-h period all the methods of wrapping significantly disrupted the host-finding process of blowflies to dismembered carcasses, with a delay of initial contact and oviposition of 30+h (dependant on wrapping) and even more in wet conditions (48+ h). Egg numbers were also reduced by as much as 99.1% on wrapped samples compared to unwrapped. These new findings highlight the importance of applying adjustments to minPMI calculations when encountering wrapped remains. Advances in the accuracy of minPMI calculations will prevent the waste of valuable police time and resources and better focus the search for witnesses and suspects in homicide investigations.

Effects of mosquito control using the microbial agent Bacillus thuringiensis israelensis (Bti) on aquatic and terrestrial ecosystems: a systematic review.

BACKGROUND: The bacterium Bacillus thuringiensis serovar israelensis (Bti) is commercially produced in various formulations for use as a larvicide worldwide, targeting especially the aquatic larval stage of mosquitoes. However, there is a concern that repeated Bti treatments may have both direct and indirect impacts on non-target organisms (NTOs) and the ecosystems they inhabit. This review evaluates the evidence for such impacts. METHODS: Literature was searched using six bibliographic databases, two search engines, and on specialist web sites. Eligibility screening was performed in two steps on (1) title/abstract, with consistency among reviewers assessed by double-screening 557 articles and (2) full text. Articles included after full text screening were critically appraised independently by two reviewers. Disagreements were reconciled through discussions. Key parameters of included studies are presented in narrative synthesis tables, including risk of bias assessments. Meta-analyses comparing treated with untreated ecosystems and using either the raw mean difference or log response ratio as effect size were performed. REVIEW FINDINGS: Ninety-five articles covering 282 case studies were included in the review. From these, we identified 119 different response variables, which were divided into nine outcome categories. Most studies investigated NTO abundance or life history (reproduction related outcomes), but diversity and community composition are also well represented as outcome categories. The studies are highly variable in methodology, rigor, and spatio-temporal scale, spanning 1 day to 21 years and from < 1m2 to > 10,000 m2. Our metanalyses revealed a consistent negative effect of Bti treatment on abundances of Chironomidae and Crustacea, and also on chironomid emergence, although from a more restricted set of studies and regions. For most remaining response variables, we judged meta-analysis unfeasible, due to low study numbers or insufficient reporting of methods and results. CONCLUSIONS: There is now a significant body of studies documenting effects of mosquito control using Bti on NTOs or other ecosystem properties, especially associated with negative effects on Chironomidae, as apparent from our meta-analyses. Accordingly, we suggest the potential for negative NTO or other ecosystem effects of Bti treatment should not be discounted a priori. Once a decision to proceed with Bti treatment has been taken, priority should be given to a well-designed program of ongoing monitoring and assessment. The paucity of rigorous studies conducted with low bias risk for most response variables undermines our capacity for evaluating how common many of the effects documented might be. Future research would benefit from a rigorous and well-replicated approach to studying Bti impacts in semi-field mesocosms or in the field, combined with a greater rigor in reporting key methodological details. A greater focus is needed on understanding the environmental factors which regulate the wider effects of mosquito control using Bti on NTOs and ecosystems, to enhance our capacity for predicting where and when Bti is most likely to have additional, negative and indirect ecological impacts.

Multimodal synergisms in host stimuli drive landing response in malaria mosquitoes.

Anopheles mosquitoes transmit malaria, which affects one-fifth of the world population. A comprehensive understanding of mosquito behaviour is essential for the development of novel tools for vector control and surveillance. Despite abundant research on mosquito behaviour, little is known on the stimuli that drive malaria vectors during the landing phase of host-seeking. Using behavioural assays with a multimodal step approach we quantified both the individual and the combined effect of three host-associated stimuli in eliciting landing in Anopheles coluzzii females. We demonstrated that visual, olfactory and thermal sensory stimuli interact synergistically to increase the landing response. Furthermore, if considering only the final outcome (i.e. landing response), our insect model can bypass the absence of either a thermal or a visual stimulus, provided that at least one of these is presented simultaneously with the olfactory stimuli, suggesting that landing is the result of a flexible but accurate stimuli integration. These results have important implications for the development of mosquito control and surveillance tools.

Understanding mosquito host-choice behaviour: a new and low-cost method of identifying the sex of human hosts from mosquito blood meals.

BACKGROUND: Mosquito-borne diseases are a global health problem, causing hundreds of thousands of deaths per year. Pathogens are transmitted by mosquitoes feeding on the blood of an infected host and then feeding on a new host. Monitoring mosquito host-choice behaviour can help in many aspects of vector-borne disease control. Currently, it is possible to determine the host species and an individual human host from the blood meal of a mosquito by using genotyping to match the blood profile of local inhabitants. Epidemiological models generally assume that mosquito biting behaviour is random; however, numerous studies have shown that certain characteristics, e.g. genetic makeup and skin microbiota, make some individuals more attractive to mosquitoes than others. Analysing blood meals and illuminating host-choice behaviour will help re-evaluate and optimise disease transmission models. METHODS: We describe a new blood meal assay that identifies the sex of the person that a mosquito has bitten. The amelogenin locus (AMEL), a sex marker located on both X and Y chromosomes, was amplified by polymerase chain reaction in DNA extracted from blood-fed Aedes aegypti and Anopheles coluzzii. RESULTS: AMEL could be successfully amplified up to 24 h after a blood meal in 100% of An. coluzzii and 96.6% of Ae. aegypti, revealing the sex of humans that were fed on by individual mosquitoes. CONCLUSIONS: The method described here, developed using mosquitoes fed on volunteers, can be applied to field-caught mosquitoes to determine the host species and the biological sex of human hosts on which they have blood fed. Two important vector species were tested successfully in our laboratory experiments, demonstrating the potential of this technique to improve epidemiological models of vector-borne diseases. This viable and low-cost approach has the capacity to improve our understanding of vector-borne disease transmission, specifically gender differences in exposure and attractiveness to mosquitoes. The data gathered from field studies using our method can be used to shape new transmission models and aid in the implementation of more effective and targeted vector control strategies by enabling a better understanding of the drivers of vector-host interactions.

Malaria hotspots explained from the perspective of ecological theory underlying insect foraging.

Hotspots constitute the major reservoir for residual malaria transmission, with higher malaria incidence than neighbouring areas, and therefore, have the potential to form the cornerstone for successful intervention strategies. Detection of malaria hotspots is hampered by their heterogenous spatial distribution, and the laborious nature and low sensitivity of the current methods used to assess transmission intensity. We adopt ecological theory underlying foraging in herbivorous insects to vector mosquito host seeking and modelling of fine-scale landscape features at the village level. The overall effect of environmental variables on the density of indoor mosquitoes, sporozoite infected mosquitoes, and malaria incidence, was determined using generalized linear models. Spatial analyses were used to identify hotspots for malaria incidence, as well as malaria vector density and associated sporozoite prevalence. We identify household occupancy and location as the main predictors of vector density, entomological inoculation rate and malaria incidence. We propose that the use of conventional vector control and malaria interventions, integrated with their intensified application targeting predicted hotspots, can be used to reduce malaria incidence in endemic and residual malaria settings.

Shady business: understanding the spatial ecology of exophilic Anopheles mosquitoes.

BACKGROUND: Understanding the ecology of exophilic anophelines is a key step toward developing outdoor control strategies to complement existing indoor control tools against malaria vectors. This study was conducted to assess the movement pattern of exophilic Anopheles mosquitoes between blood meal sources and resting habitats, and the landscape factors dictating their resting habitat choice. RESULTS: Resting clay pots were placed at 5 m, 25 m, 50 m, 75 m and 100 m away from isolated focal houses, radiating from them in four directions. The locations of the clay pots represent heterogeneous land cover types at a relatively fine spatial scale in the landscape. The effect of the landscape characters on the number of both female and male anophelines caught was modelled using zero-inflated negative binomial regression with a log link function. A total of 420 Anopheles mosquitoes (353 females and 67 males) belonging to three species; Anopheles arabiensis, Anopheles pharoensis, and Anopheles tenebrosus were caught in the resting clay pots, with An. arabiensis being the dominant species. Canopy cover, distance from the house, and land cover type were the significant landscape characters influencing the aggregation of resting mosquitoes. Both the count and binary models showed that canopy cover was the strongest predictor variable on the counts and the presence of Anopheles mosquitoes in the clay pots. Female Anopheles were most frequently found resting in the pots placed in banana plantations, and at sampling points that were at the greater distances (75 m and 100 m) from the focal house. CONCLUSIONS: This study showed that exophilic Anopheles mosquitoes tend to rest in shaded areas some distance away from human habitation. These findings are important when targeting mosquitoes outdoors, complementing the existing effort being made to control malaria vectors indoors.

Parasitism of Two Spodoptera spp. by Microplitis prodeniae (Hymenoptera: Braconidae).

Early instar larvae of the tobacco cutworm Spodoptera litura (Lepidoptera: Noctuidae) and the beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) are recognized hosts of the parasitic wasp Microplitis prodeniae Rao and Kurian (Hymenoptera: Braconidae), although M. prodeniae has previously been regarded as monophagous. In this study, we found the immature period and longevity of M. prodeniae developing in S. exigua was similar to that in S. litura. It was shown that the development time of M. prodeniae in S. exigua was 15.1 ± 0.3 d, not significantly different from 15.0 ± 0.2 d in S. litura. The parasitism rate of M. prodeniae attacking S. exigua was significantly lower than on S. litura (65.48 ± 2.29 and 43.83 ± 2.20%, respectively), whilst the female ratio of the wasp's offspring was not significantly different when developing on the two species. M. prodeniae females prefer to oviposit on the second- and third-instar host larvae of S. exigua, rather than other instars. The effects of development of M. prodeniae on two important lepidopterous pests are discussed.

Grass Pollen Affects Survival and Development of Larval Anopheles arabiensis (Diptera: Culicidae).

Nutrients in breeding sites are critical for the survival and development of malaria mosquitoes, having a direct impact on vectorial capacity. Yet, there is a limited understanding about the natural larval diet and its impact on the individual fitness of mosquitoes. Recent studies have shown that gravid Anopheles arabiensis Patton (Diptera: Culicidae) are attracted by and oviposit in grass-associated habitats. The pollen provided by these grasses is a potential source of nutrients for the larvae. Here, we assess the effect of Typha latifolia L. (Poales: Typhaceae), Echinochloa pyramidalis Lamarck, Pennisetum setaceum Forsskål, and Zea mays L. pollen on larval survival and rate of development in An. arabiensis under laboratory conditions. In addition, we characterize the carbon to nitrogen ratio and the size of pollen grains as a measure of diet quality. Carbon-rich pollen with a small grain size (T. latifolia and P. setaceum; 9.7 ± 0.3 × 103 and 5.5 ± 0.2 × 104 µm3, respectively) resulted in enhanced rates of development of An. arabiensis. In contrast, the larva fed on the nitrogen-rich control diet (TetraMin) was slower to develop, but demonstrated the highest larval survival. Larvae fed on carbon-rich and large-grained Z. mays pollen (4.1 ± 0.2 × 105 µm3) survived at similar levels as those fed on the control diet and also took a longer time to develop compared with larvae fed on the other pollens. While males and females did not appear to develop differently on the different pollen diets, males consistently emerged faster than their female counterparts. These results are discussed in relation to integrated vector management.

The role of grass volatiles on oviposition site selection by Anopheles arabiensis and Anopheles coluzzii.

BACKGROUND: The reproductive success and population dynamics, of Anopheles malaria mosquitoes is strongly influenced by the oviposition site selection of gravid females. Mosquitoes select oviposition sites at different spatial scales, starting with selecting a habitat in which to search. This study utilizes the association of larval abundance in the field with natural breeding habitats, dominated by various types of wild grasses, as a proxy for oviposition site selection by gravid mosquitoes. Moreover, the role of olfactory cues emanating from these habitats in the attraction and oviposition stimulation of females was analysed. METHODS: The density of Anopheles larvae in breeding sites associated with Echinochloa pyramidalis, Echinochloa stagnina, Typha latifolia and Cyperus papyrus, was sampled and the larvae identified to species level. Headspace volatile extracts of the grasses were collected and used to assess behavioural attraction and oviposition stimulation of gravid Anopheles arabiensis and Anopheles coluzzii mosquitoes in wind tunnel and two-choice oviposition assays, respectively. The ability of the mosquitoes to differentiate among the grass volatile extracts was tested in multi-choice tent assays. RESULTS: Anopheles arabiensis larvae were the most abundant species found in the various grass-associated habitats. The larval densities described a hierarchical distribution, with Poaceae (Echinochloa pyramidalis and Echinochloa stagnina)-associated habitat sites demonstrating higher densities than that of Typha-associated sites, and where larvae were absent from Cyperus-associated sites. This hierarchy was maintained by gravid An. arabiensis and An. coluzzii mosquitoes in attraction, oviposition and multi-choice assays to grass volatile extracts. CONCLUSIONS: The demonstrated hierarchical preference of gravid An. coluzzii and An. arabiensis for grass volatiles indicates that vegetation cues associated with larval habitats are instrumental in the oviposition site choice of the malaria mosquitoes. Identifying volatile cues from grasses that modulate gravid malaria mosquito behaviours has distinct potential for the development of tools to be used in future monitoring and control methods.

Plant-Mediated Effects on Mosquito Capacity to Transmit Human Malaria.

The ecological context in which mosquitoes and malaria parasites interact has received little attention, compared to the genetic and molecular aspects of malaria transmission. Plant nectar and fruits are important for the nutritional ecology of malaria vectors, but how the natural diversity of plant-derived sugar sources affects mosquito competence for malaria parasites is unclear. To test this, we infected Anopheles coluzzi, an important African malaria vector, with sympatric field isolates of Plasmodium falciparum, using direct membrane feeding assays. Through a series of experiments, we then examined the effects of sugar meals from Thevetia neriifolia and Barleria lupilina cuttings that included flowers, and fruit from Lannea microcarpa and Mangifera indica on parasite and mosquito traits that are key for determining the intensity of malaria transmission. We found that the source of plant sugar meal differentially affected infection prevalence and intensity, the development duration of the parasites, as well as the survival and fecundity of the vector. These effects are likely the result of complex interactions between toxic secondary metabolites and the nutritional quality of the plant sugar source, as well as of host resource availability and parasite growth. Using an epidemiological model, we show that plant sugar source can be a significant driver of malaria transmission dynamics, with some plant species exhibiting either transmission-reducing or -enhancing activities.

Imperfection works: Survival, transmission and persistence in the system of Heliothis virescens ascovirus 3h (HvAV-3h), Microplitis similis and Spodoptera exigua.

Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis.

24-h sheltering behaviour of individually kept horses during Swedish summer weather.

BACKGROUND: Provision of shelter for horses kept on summer pasture is rarely considered in welfare guidelines, perhaps because the benefits of shelter in warm conditions are poorly documented scientifically. For cattle, shade is a valued resource during summer and can mitigate the adverse effects of warm weather on well-being and performance. We found in a previous study that horses utilized shelters frequently in summer. A shelter with a roof and closed on three sides (shelter A) was preferred and can reduce insect pressure whereas a shelter with roof and open on three sides was not utilized. However, shelter A restricts the all-round view of a horse, which may be important for horses as flight animals. Therefore, we studied whether a shelter with roof, where only the upper half of the rear wall was closed (shelter B), would be utilized while maintaining insect protection properties and satisfying the horses' sense for security. A third shelter was offered with walls but no roof (shelter C) to evaluate whether the roof itself is an important feature from the horse's perspective. Eight Warmblood horses were tested each for 2 days, kept individually for 24 h in two paddocks with access to shelters A and B, or shelters A and C, respectively. Shelter use was recorded continuously during the night (1800-2400 h, 0200-0600 h) and the following day (0900-1600 h), and insect defensive behaviour (e.g., tail swish) in instantaneous scan samples at 5-min intervals during daytime. RESULTS: Seven horses used both shelters A and B, but when given the choice between shelters A and C, shelter C was scarcely visited. There was no difference in duration of shelter use between night (105.8 ± 53.6 min) and day (100.8 ± 53.8, P = 0.829). Daytime shelter use had a significant effect on insect defensive behaviours (P = 0.027). The probability of performing these behaviours was lowest when horses used shelter A compared to being outside (P = 0.038). CONCLUSIONS: Horses only utilized shelters with a roof whilst a shelter with roof and closed on three sides had the best potential to lower insect disturbance during daytime in summer.

Role of glucosinolates in insect-plant relationships and multitrophic interactions.

Glucosinolates present classical examples of plant compounds affecting insect-plant interactions. They are found mainly in the family Brassicaceae, which includes several important crops. More than 120 different glucosinolates are known. The enzyme myrosinase, which is stored in specialized plant cells, converts glucosinolates to the toxic isothiocyanates. Insect herbivores may reduce the toxicity of glucosinolates and their products by excretion, detoxification, or behavioral adaptations. Glucosinolates also affect higher trophic levels, via reduced host or prey quality or because specialist herbivores may sequester glucosinolates for their own defense. There is substantial quantitative and qualitative variation between plant genotypes, tissues, and ontogenetic stages, which poses specific challenges to insect herbivores. Even though glucosinolates are constitutive defenses, their levels are influenced by abiotic and biotic factors including insect damage. Plant breeders may use knowledge on glucosinolates to increase insect resistance in Brassica crops. State-of-the-art techniques, such as mutant analysis and metabolomics, are necessary to identify the exact role of glucosinolates.

Combined effect of intercropping and turnip root fly (Delia floralis) larval feeding on the glucosinolate concentrations in cabbage roots and foliage.

The effects of plant competition and herbivory on glucosinolate concentrations in cabbage root and foliage were investigated in a cabbage-red clover intercropping system. Cabbage plants were grown under different competitive pressures and with varying degrees of attack by root-feeding Delia floralis larvae. Glucosinolate concentrations in cabbage were affected both by intercropping and by D. floralis density. Glucosinolate concentrations in foliage generally decreased as a response to intercropping, while the responses to insect root damage of individual glucosinolates were weaker. Root glucosinolates responded more strongly to both intercropping and egg density. Total root glucosinolate concentration decreased with clover density, but only at high egg densities. Increased egg density led to opposite reactions by the indole and aliphatic glucosinolates in roots. The responses of individual root glucosinolates to competition and root damage were complex and, on occasion, nonlinear. Reduced concentrations of several glucosinolates and the tendency towards a decrease in total concentration in cabbage foliage caused by intercropping and larval damage suggest that competing plants or plants with root herbivory do not allocate the same resources as unchallenged plants towards sustaining levels of leaf defensive compounds. This could also be true for root glucosinolate concentrations at high egg densities. In addition, the results suggest that changes occurring within a structural group of glucosinolates may be influenced by changes in a single compound, e.g., glucobrassicin (indol-3-ylmethyl) in foliage or sinigrin (2-propenyl) in roots.

The pollen beetle, Meligethes aeneus, changes egg production rate to match host quality.

Motivation-based models dominate current theory concerning host plant selection for oviposition by herbivorous insects. A female searching for a host plant will be more likely to accept a host which is of inferior quality for her offspring if motivation is high, e.g. a large eggload or long time since last oviposition. This implies that insects will accumulate eggs if exposed to hosts of low acceptability and after a time lay eggs on such hosts. An alternative strategy for insects when exposed to less acceptable hosts is to stop producing, instead of accumulating, eggs. Thus, resources would be saved until a more acceptable host is found. If this hypothesis is true, a herbivorous insect would cease egg production when exposed to hosts of low acceptability and resume egg production when exposed to hosts of high acceptability. Previous exposure should not affect oviposition rate when an insect encounters a new host of a different quality. In an earlier study pollen beetles, Meligethes aeneus (F.) (Coleoptera: Nitidulidae), did not accumulate eggs in the absence of high quality hosts. In this study we monitored the daily oviposition rate of female pollen beetles on hosts plants of low (Sinapis alba L.), intermediate (Brassica nigra Koch) or high (B. napus L.) acceptability over a 5-day period. Individuals were then switched to an oviposition resource of a different acceptability. Beetles moved from high- to low-acceptability plants reduced their oviposition rate considerably. In the opposite case, low to high acceptability, the rate of oviposition increased markedly after the switch. When M. aeneus were moved from the high-acceptability host to that of intermediate acceptability oviposition rate was modified accordingly. However, when moved to the intermediate host from a host of low acceptability oviposition on B. nigra was much less than would normally be expected. A possible mechanism for this finding is discussed. M. aeneus, by adjusting oviposition rate to host acceptability, maximizes the average host quality for offspring, even at the cost of a lower egg-laying rate.