The field of ecological immunology currently relies on using a number of immune effectors or markers. These markers are usually used to infer ecological trade-offs (via conflicts in resource allocation), though physiological nature of these markers remains elusive. Here, we review markers frequently used in insect evolutionary ecology research: cuticle darkening, haemocyte density, nodule/capsule formation, phagocytosis and encapsulation/melanization via use of nylon filaments and beads, phenoloxidase activity, nitric oxide production, lysozyme and antimicrobial peptide production. We also provide physiologically based information that may shed light on the probable trade-offs inferred when these markers are used. In addition, we provide a number of methodological suggestions to improve immune marker assessment.

Effective, statistically robust sampling and surveillance strategies form an integral component of large agricultural industries such as the grains industry. Intensive in-storage sampling is essential for pest detection, integrated pest management (IPM), to determine grain quality and to satisfy importing nation's biosecurity concerns, while surveillance over broad geographic regions ensures that biosecurity risks can be excluded, monitored, eradicated or contained within an area. In the grains industry, a number of qualitative and quantitative methodologies for surveillance and in-storage sampling have been considered. Primarily, research has focussed on developing statistical methodologies for in-storage sampling strategies concentrating on detection of pest insects within a grain bulk; however, the need for effective and statistically defensible surveillance strategies has also been recognised. Interestingly, although surveillance and in-storage sampling have typically been considered independently, many techniques and concepts are common between the two fields of research. This review aims to consider the development of statistically based in-storage sampling and surveillance strategies and to identify methods that may be useful for both surveillance and in-storage sampling. We discuss the utility of new quantitative and qualitative approaches, such as Bayesian statistics, fault trees and more traditional probabilistic methods and show how these methods may be used in both surveillance and in-storage sampling systems.

Aphids are major pests in apple orchards, debilitating the crop and spreading disease. We investigated whether early-season predation by canopy spiders may be effectively controlling aphid numbers in three organic orchards. For this purpose, we monitored the aphid population dynamics from the winter eggs to colony stages and compared this to spider abundances and rates of predation on aphids detected by diagnostic polymerase chain reaction. For the latter, we applied existing general aphid primers. We found that spiders ate colony fundatrices and that aphid numbers were negatively related to spider abundance. Spiders were the main active predators within the orchards when the first colony fundatrices were present, indicating their importance in the early control of aphid populations.

Interpreting spatial patterns in the abundance of species over time is a fundamental cornerstone of ecological research. For many species, this type of analysis is hampered by datasets that contain a large proportion of zeros, and data that are overdispersed and spatially autocorrelated. This is particularly true for insects, for which abundance data can fluctuate from zero to many thousands in the space of weeks. Increasingly, an understanding of the ways in which environmental variation drives spatial and temporal patterns in the distribution, abundance and phenology of insects is required for management of pests and vector-borne diseases. In this study, we combine the use of smoothing techniques and generalised linear mixed models to relate environmental drivers to key phenological patterns of two species of biting midges, Culicoides pulicaris and C. impunctatus, of which C. pulicaris has been implicated in transmission of bluetongue in Europe. In so doing, we demonstrate analytical tools for linking the phenology of species with key environmental drivers, despite using a relatively small dataset containing overdispersed and zero-inflated data. We demonstrate the importance of landcover and climatic variables in determining the seasonal abundance of these two vector species, and highlight the need for more empirical data on the effects of temperature and precipitation on the life history traits of palearctic Culicoides spp. in Europe.

Transgenic (Bt) cotton dominates Australian cotton production systems. It is grown to control feeding damage by lepidopteran pests such as Helicoverpa armigera. The possibility that these moths might become resistant to Bt remains a threat. Consequently, refuge crops (with no Bt) must be grown with Bt cotton to produce large numbers of Bt-susceptible moths to reduce the risk of resistance developing. A key assumption of the refuge strategy, that moths from different host plant origins mate at random, remains untested. During the period of the study reported here, refuge crops included pigeon pea, conventional cotton (C3 plants), sorghum or maize (C4 plants). To identify the relative contributions made by these (and perhaps other) C3 and C4 plants to populations of H. armigera in cotton landscapes, we measured stable carbon isotopes (δ13C) within individual moths captured in the field. Overall, 53% of the moths were of C4 origin. In addition, we demonstrated, by comparing the stable isotope signatures of mating pairs of moths, that mating is indeed random amongst moths of different plant origins (i.e. C3 and C4). Stable nitrogen isotope signatures (δ15N) were recorded to further discriminate amongst host plant origins (e.g. legumes from non-legumes), but such measurements proved generally unsuitable. Since 2010, maize and sorghum are no longer used as dedicated refuges in Australia. However, these plants remain very common crops in cotton production regions, so their roles as ‘unstructured’ refuges seem likely to be significant.

Anthropogenic range expansion and cultural practices have modified the distribution, abundance and genetic diversity of domesticated organisms, thereby altering multitrophic assemblages through space and time. The putative Mesoamerican domestication centre of the common bean, Phaseolus vulgaris L., in Mexico allows investigating the effects of plant domestication on the genetic structure of members of a multitrophic system. The aim of this study was to compare the evolutionary history of Horismenus parasitoids (Hymenoptera: Eulophidae) to those of their bruchid beetle hosts (Coleoptera: Bruchidae) and their domesticated host plant (P. vulgaris), in the context of traditional agriculture in Mexico. We analyzed the population genetic structure of four Horismenus species in Mexico using mitochondrial COI haplotype data. The two most abundant parasitoid species were Horismenus depressus and Horismenus missouriensis. Horismenus missouriensis were infected by Wolbachia endosymbionts and had little to no population differentiation (FST = 0.06). We suspect the mitochondrial history of H. missouriensis to be blurred by Wolbachia, because differentiation among infected vs. non-infected individuals exists (FST = 0.11). Populations of H. depressus were found to be highly differentiated (FST = 0.34), but the genetic structuring could not be explained by tested spatial components. We then compared the genetic structure observed in this parasitoid species to previously published studies on bruchid beetles and their host plants. Despite extensive human-mediated migration and likely population homogenization of its two Acanthoscelides bruchid beetle hosts, H. depressus populations are structured like its host plant, by a recent dispersal from a diverse ancestral gene pool. Distinct evolutionary dynamics may explain inconsistent patterns among trophic levels. Parasitoids likely migrate from wild bean populations and are poorly adapted to bean storage conditions similar to their bruchid beetle hosts. Integrating several trophic levels to the study of evolutionary history has proven to be fruitful in detecting different ecological responses to human-mediated disturbances and host parasite interactions.

To advance our restricted knowledge on mosquito biodiversity and distribution in Belgium, a national inventory started in 2007 (MODIRISK) based on a random selection of 936 collection points in three main environmental types: urban, rural and natural areas. Additionally, 64 sites were selected because of the risk of importing a vector or pathogen in these sites. Each site was sampled once between May and October 2007 and once in 2008 using Mosquito Magnet Liberty Plus traps. Diversity in pre-defined habitat types was calculated using three indices. The association between species and environmental types was assessed using a correspondence analysis. Twenty-three mosquito species belonging to traditionally recognized genera were found, including 21 indigenous and two exotic species. Highest species diversity (Simpson 0.765) and species richness (20 species) was observed in natural areas, although urban sites scored also well (Simpson 0.476, 16 species). Four clusters could be distinguished based on the correspondence analysis. The first one is related to human modified landscapes (such as urban, rural and industrial sites). A second is composed of species not associated with a specific habitat type, including the now widely distributed Anopheles plumbeus. A third group includes species commonly found in restored natural or bird migration areas, and a fourth cluster is composed of forest species. Outcomes of this study demonstrate the effectiveness of the designed sampling scheme and support the choice of the trap type. Obtained results of this first country-wide inventory of the Culicidae in Belgium may serve as a basis for risk assessment of emerging mosquito-borne diseases.

Two Macrolophus species, M. melanotoma (=M. caliginosus) and M. pygmaeus, have been referred to as efficient predators of several key pests on vegetable crops in Europe. However, due to the great morphological similarity of these species, they have been confused, with important consequences for inoculative releases of these predators in greenhouses and for the conservation of their natural populations on greenhouse and outdoor crops. In this work, we developed tools to identify these morphologically very similar species. We first confirmed the specific status of two Macrolophus populations collected on their respective host plants (Dittrichia viscosa and tomato) through crossing experiments. Then, using multivariate morphometric analysis, we proposed a linear discriminant function that combines head measurements separating males from the two species without error. Finally, we designed specific primers for a mitochondrial DNA region that were able to distinguish field-collected Macrolophus individuals through conventional PCR. In conclusion, the tools developed in the present study will allow reliable identification of the Macrolophus species present in crops and in the native flora that are the source of populations that colonise them. They will also allow correct identification of mass reared Macrolophus to be introduced in greenhouse crops in inoculative releases.

Host specificity of Neotropical hard ticks (Acari: Ixodidae) was analyzed by using the number of hosts species for each tick species and the index of host specificity STD*, which integrates phylogenetic and ecological information. The analyses were based on 4172 records of hard ticks collected from wild and domestic tetrapods. Most tick species included in this study were associated with three to 20 host species. No tick species has been associated either with a single species or with a single genus of host. It was found that the number of host species is sensitive to sampling effort, but not the STD*. The most frequent values of STD* were between 2.5 and 3.5, which shows that the host species more frequently used by Neotropical hard tick species belong to different families or different orders. Immature stages tend to use a broader taxonomic range of hosts than adults, and the interpretation of both measures of host specificity used in this study led to the conclusion that the impact of non-endemic hosts does not alter the patterns of host specificity in Neotropical hard ticks. The index STD* showed that a high proportion of tick species has phylogenetically unrelated species as principal hosts. The conclusion reached in this work indicates that strict host specificity is not common among Neotropical hard ticks and suggests that the influence of tick ecology and evolution of habitat specificity, tick generation time, phenology, time spent off the host and the type of life-cycle could be more important than hosts species.

Arion vulgaris Moquin-Tandon 1855 is one of the most important invasive species in Europe, affecting both biodiversity and agriculture. The species is spreading in many parts of Europe, inflicting severe damage to horticultural plants and cultivated crops partly due to a lack of satisfactory and effective management solutions. Molluscicides have traditionally been used to manage slug densities, although the effects are variable and some have severe side-effects on other biota. Thus, there is a need to explore potential alternatives such as biological control. The nematode Phasmarhabditis hermaphrodita is the only biological agent that has been applied commercially so far. However, other biological control agents such as carabid beetles have also been found to be promising. In addition, some carabid species have been shown to feed on A. vulgaris in the field as well as in the laboratory. Two species in particular have been found to be important predators of A. vulgaris, and these species are also common in agricultural environments: Pterostichus melanarius and Carabus nemoralis. This study is the first to use semi-field experiments in a strawberry field, manipulating densities, to investigate how P. melanarius and C. nemoralis affect densities of A. vulgaris eggs and juveniles, respectively. Gut contents of C. nemoralis were analysed using multiplex PCR methods to detect DNA of juvenile slugs. Results show that both P. melanarius and C. nemoralis significantly affect densities of slug eggs and juvenile slugs under semi-field conditions and that C. nemoralis seems to prefer slugs smaller than one gram. Carabus nemoralis seems to be especially promising in reducing densities of A. vulgaris, and future studies should investigate the potential of using this species as a biological control agent.

Among the herbivorous arthropods that feed on strawberry, the most important are the two-spotted spider mite (TSSM), Tetranychus urticae Koch, and several species of aphids. Mites and aphids belong to different guilds that coexist in the field and feed on the undersides of strawberry leaflets. However, the occurrence of large numbers of individuals of both species on the same leaflet is rarely recorded. We hypothesize that negative interactions between TSSM and aphids explain the intraplant distribution of these herbivores. We first examined the spatial coincidence of both herbivores in the field. Under experimental conditions, we then analyzed: (i) the rate of increase of TSSM and the aphid Chaetosiphon fragaefolii (Cockerell), growing individually and together; (ii) the effect of honeydew on TSSM preference; and (iii) the effect of previous strawberry leaflet damage by TSSM on C. fragaefolii preference. The proportion of TSSM that coincided with at least one aphid decreased as the percentage of leaflets with TSSM increased. The spatial coincidence index between aphids and TSSM increased together with the percentage of TSSM-infested leaflets. TSSM showed both a lower rate of increase when they shared the same leaflet with C. fragaefolii and lower fecundity on strawberry discs with honeydew. The rate of increase of C. fragaefolii did not change on co-occupied leaves, but the aphid species moved to the other side of leaflets shared with TSSM. Negative interactions resulting in a tendency for species to avoid each other, such as demonstrated herein, can affect distribution and performance of herbivorous arthropods.

Rhynchophorus ferrugineus (Coleoptera, Curculionidae) is the most threatening pest of palms worldwide. The potential of gamma-irradiated males to spread a pathogenic strain of the entomopathogenic fungus Beauveria bassiana (Ascomycota: Clavicipitaceae) to control this pest was studied. First, the effects of gamma irradiation (15 and 25 Gy) on the mating success and performance of adult males irradiated at age one day were studied in the laboratory. Although male longevity decreased after irradiation (118.6 vs. 244.7 days for irradiated and control males, respectively) and their testes suffered from the treatment, fecundity of mated females did not depend on the irradiation status of the male (86.8 ± 5.5 eggs in 15 days). However, egg hatching was significantly lower in couples with irradiated males (31.4% vs. 86.5% for irradiated and control couples, respectively), and this value decreased after a second mating (6.1% vs. 85.9%). Therefore, irradiation did not affect male sexual competiveness but sperm quality. Second, a semi-field assay was carried out to evaluate infestation in young Phoenix canariensis caused by different combinations of couples with irradiated and/or B. bassiana-challenged males. The number of immature stages found in infested palms was significantly higher when females mated with untreated males and lower when mated with irradiated males (either B. bassiana-infected or not). Some females from the fungus-challenged treatments showed post-mortem hyphal growth, and this horizontal transmission proves that irradiated males could act as a vector for B. bassiana and should be considered as a new method to improve the biological control of R. ferrugineus.