The feasibility and efficacy of early-season releases of a generalist predator (Forficula auricularia L.) to control populations of the RAA (Dysaphis plantaginea Passerini) in Southeastern France.

Augmentative biological control is not commonly used in commercial orchards. We used an exclusion system to evaluate the potential of early-season releases of the European earwig (Forficula auricularia L., Dermaptera: Forficulidae) for control of the rosy apple aphid (Dysaphis plantaginea Passerini, Hemiptera: Aphididae) in the spring of 2009 in two pesticide-free apple orchards. In order to conduct this experiment we successfully reared earwigs with a high survival rate of nymphs (more than 96%) which may have commercial application. There were three treatments in the study: (i) a 'release treatment' where we confined the released earwigs in the canopy by using a barrier system; (ii) an 'exclusion treatment' where we blocked free access of earwigs into the canopy using the same barrier system; and (iii) a 'control treatment' that represented the natural situation. Contrary to expectations, earwig releases did not reduce D. plantaginea populations. In general, the abundance of natural enemies and their groups did not differ significantly among treatments, except for earwigs. We observed that the exclusion systems we used successfully kept both earwigs and ants away from tree canopies; total numbers on trees in the 'exclusion treatment' were significantly lower than on the other two treatments. Due to the complexity and difficulty of evaluating augmentative releases of natural enemies in open orchard conditions, we conclude that new technical approaches to control site conditions are needed when conducting such studies.

Comparing the genetic structure of codling moth Cydia pomonella (L.) from Greece and France: long distance gene-flow in a sedentary pest species.

Codling moth Cydia pomonella L. (Lepidoptera: Tortricidae) is the most important insect pest of apple production in Europe. Despite the economic importance of this pest, there is not information about the genetic structure of its population in Greece and the patterns of gene-flow which might affect the success of control programs. In this study, we analysed nine samples from apple, pear and walnut from various regions of mainland Greece using 11 microsatellite loci. Six samples from the aforementioned hosts from southern France were also examined for comparison. Bayesian clustering and genetic distance analyses separated the codling moth samples in two genetic clusters. The first cluster consisted mainly of the individuals from Greece, and the second of those from France, although admixture and miss-classified individuals were also observed. The low genetic differentiation among samples within each country was also revealed by F(ST) statistics (0.009 among Greek samples and 0.0150 among French samples compared to 0.050 global value among all samples and 0.032 the mean of the pair-wise values between the two countries). These F(ST) values suggest little structuring at large geographical scales in agreement with previous published studies. The host species and local factors (climatic conditions, topography, pest control programs) did not affect the genetic structure of codling moth populations within each country. The results are discussed in relation to human-made activities that promote gene-flow even at large geographic distances. Possible factors for the genetic differentiation between the two genetic clusters are also discussed.

Tools for resistance monitoring in oriental fruit moth (Lepidoptera: Tortricidae) and first assessment in Brazilian populations.

In southern Brazilian apple (Malus spp.) orchards, predominantly organophosphates are used to control the oriental fruit moth, Cydia molesta (Busck) (Lepidoptera: Tortricidae), but control failures often occur. Therefore the susceptibility of three C. molesta Brazilian populations was investigated to five insecticides of different groups and modes of action, in comparison with a susceptible laboratory strain mass reared in southern France for >10 yr. At the same time, comparative biochemical and genetic analysis were performed, assessing the activities of the detoxification enzymatic systems and sequencing a gene of insecticide molecular target to find out markers associated with resistance. The three Brazilian populations were significantly resistant to chlorpyrifos ethyl compared with the reference strain. One of the field populations that had been frequently exposed to deltamethrin treatments showed significant decreasing susceptibility to this compound, whereas none of the three populations had loss of susceptibility to tebufenozide and thiacloprid compared with the reference strain. All three populations had slight but significant increases of glutathione transferase and carboxylesterases activities and significant decrease of specific acetylcholinesterase activities compared with the reference. Only the most resistant population to chlorpyriphos exhibited a significantly higher mixed function oxidase activity than the reference. The acetylcholinesterase of females was significantly less inhibited by carbaryl in the Brazilian populations than in the reference strain (1.7-2.5-fold), and this difference was not expressed in the male moth. However, no mutation in the MACE locus was detected. These biological and molecular characterizations of adaptive response to insecticides in C. molesta provide tools for early detection of insecticide resistance in field populations of this pest.

Worldwide variability of insecticide resistance mechanisms in the codling moth, Cydia pomonella L. (Lepidoptera: Tortricidae).

The activity of detoxifying enzymes (glutathione-S-transferases (GST), mixed-function oxidases (MFO), and esterases (EST)) and the presence of insensitive variants of target proteins (sodium channel and acetylcholinesterase) were examined in individual male and female codling moths. Twenty-nine populations from 11 countries and two laboratory strains were examined. Populations were classified as either unsprayed or sprayed. The ranges of enzyme activities across field populations varied 15-fold, 485-fold and fourfold for GST, MFO and EST, respectively. MFO was the only enzyme whose activity differed in a binomial classification of orchards based on their spray history. Few differences in enzyme activities were found due to sex among populations; and, in these cases, males had higher GST and lower MFO and EST activities than females. Activities of the three enzymatic systems across all populations were positively correlated. Populations from Greece, Argentina and Uruguay had significant percentages of moths with elevated GST and MFO activities. The co-occurrence of moths expressing both elevated MFO and low EST activities was found in conventional orchards from the Czech Republic and France. Chile was the only country where populations from treated orchards did not include a significant proportion of individuals with enhanced enzyme activity. The kdr mutation was found at significant levels in ten populations from five countries, including all French and Argentinean populations. The mutation in AChE was only detected in the Spanish population.

Genetic architecture in codling moth populations: comparison between microsatellite and insecticide resistance markers.

The codling moth, Cydia pomonella, is renowned for developing resistance to insecticides and causing significant economic damage to pome fruits worldwide. In spite of its economic importance, little is known about the patterns of movement of this pest and the effects of insecticide treatment on the population genetic structure. Here, we investigated the genetic structure of the pest in 27 orchards from France, Italy, Armenia and Chile at seven microsatellite loci and two resistance markers [biochemical activity of cytochrome P450 oxidases and proportion of knockdown resistance (kdr) alleles in the sodium channel gene]. According to the microsatellite loci, we detected isolation by distance at the supranational scale but found no evidence of geographical structure among the 24 French orchards, which were mainly structured by the intensity of the insecticide treatments. Similarly, the highest levels of metabolic resistance associated with activity of the cytochrome P450 oxidases were detected in the most treated orchards. The kdr alleles were observed in southern France and Armenia where the pyrethroid insecticides were or have been intensively sprayed. The intensity of the insecticide treatments marginally affected the allelic richness in each orchard, but not the level of inbreeding. These results suggest important and high-distance gene flow among the codling moth populations, which were mainly structured according to the history of insecticide applications. Differences in mutation-migration-drift equilibrium among treated and untreated orchards also suggest that insecticide applications are the main force regulating the local dynamics of codling moth populations.

Constraints on adaptive mutations in the codling moth Cydia pomonella (L.): measuring fitness trade-offs and natural selection.

Adaptive changes in populations encountering a new environment are often constrained by deleterious pleiotropic interactions with ancestral physiological functions. Evolutionary responses of populations can thus be limited by natural selection under fluctuating environmental conditions, if the adaptive mutations are associated with pleiotropic fitness costs. In this context, we have followed the evolution of the frequencies of insecticide-resistant mutants of Cydia pomonella when reintroduced into an untreated environment. The novel set of selective forces after removal of insecticide pressure led to the decline of the frequencies of resistant phenotypes over time, suggesting that the insecticide-adapted genetic variants were selected against the absence of insecticide (with a selective coefficient estimated at 0.11). The selective coefficients were also estimated for both the major cytochrome P450-dependent monooxygenase (MFO) and the minor glutathione S-transferase (GST) systems (0.17 and negligible, respectively), which have been previously shown to be involved in resistance. The involvement of metabolic systems acting both through xenobiotic detoxification and biosynthetic pathways of endogenous compounds may be central to explaining the deleterious physiological consequences resulting from pleiotropy of adaptive changes. The estimation of the magnitude of the fitness cost associated with insecticide resistance in C. pomonella suggests that resistance management strategies exclusively based on insecticide alternations would be unlikely to delay such a selection process.

Deltamethrin resistance in the codling moth (Lepidoptera: Tortricidae): inheritance and number of genes involved.

The inheritance of deltamethrin resistance in Cydia pomonella (L.) has been investigated by crossing a resistant (Rv) and a susceptible (Sv) strain, derived from a population collected in south-eastern France in 1995. Deltamethrin resistance was suspected to be under the control of a kdr-type allele and an enhanced mixed-function oxidase (mfo). F(1) and F(2) progenies were therefore tested through dose-response and enzyme assays. Dose-response relationships indicated that resistance was inherited as an autosomal incompletely recessive (D=-0.199) character, involving at least two genes. Enzyme measures suggested the contribution of 1.2 genes to the expression of mfo, with incomplete dominance (D=0.460). Our results support the hypothesis of a polygenic response to deltamethrin selection in the Rv strain, including a major kdr-type allele with a minor effect of mfos. In the light of these findings, we consider the resistance in codling moth populations in south-eastern France as a product of an adaptive sequential selection process, occurring through the sequential addition of resistance genes.

Is attract-and-kill technology potent against insecticide-resistant Lepidoptera?

Attract-and-kill techniques, associating an attractant and a contact insecticide in a sticky formulation, are a new way of controlling Lepidopteran pests. Insecticide resistance may, however, limit the effectiveness and even the attractiveness of such formulations where resistance pleiotropic effects influence pheromone perception. We have tested this hypothesis on resistant codling moths Cydia pomonella (L) using a commercial formulation containing (E,E)-8,10-dodecadien-1-ol (codlemone), the major component of the sex pheromone, as an attractant and permethrin as toxicant. We first compared the attractiveness of codlemone in a wind tunnel and the contact toxicity of pyrethroids on males of one susceptible and of three strains selected for resistance to diflubenzuron, deltamethrin and azinphos-methyl. The dose-response relationships of males of susceptible and resistant strains to codlemone did not differ significantly. The deltamethrin-selected strain was the most resistant to pyrethroids, exhibiting 138-, 25- and 18-fold resistance ratios to deltamethrin, cypermethrin and permethrin, respectively. The efficiency of the attracticide formulation, applied successively on filter paper support, glass support and wood support, was estimated by recording the mortality delay of males after natural contact with the formulation in the wind tunnel. The deltamethrin- and diflubenzuron-resistant strains were significantly less affected than the susceptible strain by contact with the attracticide on the wooden support, exhibiting 58- and 2.3-fold greater LT50 ratios, respectively. Mortality of deltamethrin-resistant moths did not exceed 40% after 48 h. The LT50 value was significantly greater on filter paper support than on the two other supports. Surprisingly, the LT50 ratio of the deltamethrin-resistant strain was markedly higher on filter paper support (1021-fold), which was more absorbent, than on the glass support (31-fold). No sublethal effects in terms of pheromone response, mating or fecundity occurred in moths surviving contact with the attracticide. Choice of insecticides in attracticide formulations will be influenced by the resistance background of the target pests. Principles of insecticide resistance management may also be applied to attract-and-kill technology by alternating with other insecticides or control methods.

Grain quality of rices grown in irrigated and upland cultures.

Upland culture of nine varieties at Ivory Coast resulted in decreases in grain weight, brown and milled rice translucency, Chroma meter L* of rough rice, Chroma meter L* and b* of milled rice, milled rice protein, and Amylograph peak viscosity, and in increases in total phenolics of hull and brown rice, and Chroma meter a* of rough, brown, and milled rice of 5 to 8 of the varieties as compared with irrigated culture. Upland culture had no consistent effect on grain resistance to Angoumois grain moth (Sitotraga cerealella).