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Area-wide sterile insect technique (SIT) programs assume that offspring reduction of the target population correlates with the mating success of the sterile males released. However, there is a lack of monitoring tools to prove the success of these programs in real-time. Field-cage tests were conducted under the environmental conditions of the Mediterranean coast of Spain to estimate: (a) the mating success of sterile Vienna-8 (V8) Ceratitis capitata males using molecular markers and (b) their efficacy to reduce C. capitata populations under six release ratios of wild females to wild males to V8 males (1:0:0, 1:1:0, 1:1:1, 1:1:5, 1:1:10, and 1:1:20). Statistical models were developed to predict: (a) the number of females captured in traps, (b) sperm ID (sterile or not) in spermathecae of the trapped females, and (c) the viable offspring produced, using release ratio and temperature as predictors. The number of females captured was affected by relative humidity. However, its influence in the model was low. Female captures were significantly higher in ratios 1:0:0 compared to ratios where V8 males were released. The proportion of V8 sperm in spermathecae increased with temperature and with the number of V8 males released, but leveled off between ratios 1:1:10 and 1:1:20. In all seasons, except winter (no offspring), viable offspring increased with temperature and was lowest for ratio 1:1:20. For the first time, a strong negative relationship between proportion of V8 sperm detected by molecular tools and C. capitata offspring was established. The models obtained should contribute to enhance the efficacy of SIT programs against this pest.

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Tetranychus urticae Koch is a cosmopolitan mite considered as the most polyphagous species among spider mites. This mite is a key pest of clementine mandarins in Eastern Spain, where Spanish clementine production concentrates. Crop management practices can affect the population dynamics of this mite and, consequently, its impact on the orchard. Microsatellite markers were used to study mite population genetics from two commercial orchards which had been managed differently following Integrated Pest Management (IPM) or Organic Pest Management (OPM) schemes during four consecutive years. A multiplex system including 20 microsatellite loci was designed specifically and allowed an efficient and inexpensive genotyping of individual mites. We found that the IPM population had a stronger fluctuation of population structure and higher genetic diversity compared to OPM population. Thus, our study concludes that crop management has an impact on the population genetics of T. urticae which may be related to the alternation of some acaricides under IPM.

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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.

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Tetranychus urticae Koch is a cosmopolitan phytophagous mite considered as the most polyphagous species among spider mites. Population genetic studies using molecular markers such as microsatellites have proven to be extremely informative to address questions about population structure, phylogeography and host preferences. The aim of this study was to increase the available molecular tools to gain insight into the genetic structure of T. urticae populations of citrus orchards, which might help in their management. Five microsatellite DNA libraries were developed using probes with the motifs CT, CTT, GT and CAC following the FIASCO protocol. Positive clones, those that included the insert with the microsatellite, were detected using the PIMA-PCR technique. Combinations of primers were designed on 22 out of 32 new microsatellites loci and their polymorphism was tested in four populations sampled along the eastern coast of Spain. Eleven successful amplifications were obtained. Cross amplification was tested in the tetranychids Aphlonobia histricina, Eutetranychus banksi, E. orientalis, Oligonychus perseae, Panonychus citri, Tetranychus evansi, T. okinawanus and T. turkestani, and the phytoseiids Amblyseius swirskii, A. cucumeris, A. andersoni, Euseius stipulatus, Neoseiulus barkeri, N. californicus, Phytoseiulus persimilis and Typhlodromus phialatus. Eight successful cross amplifications were obtained.

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The red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae), is an economically important pest of palms worldwide. To better understand the phenology of R. ferrugineus populations in the Mediterranean basin, the objective of the present work has been to estimate the effect of temperatures within the range 10-25°C on the reproductive parameters of R. ferrugineus and to determine its lower temperature thresholds for oviposition and egg hatching. Our results confirm that oviposition in R. ferrugineus is strongly affected by temperature. Lower thresholds for oviposition and egg hatching (15.45° and 13.95°C, respectively) are below the mean monthly temperatures registered in winter in most of the northern shore of the Mediterranean basin. Under these circumstances, new palm infestations would be difficult during most of the winter. These results should be taken into account when planning some palm management practices, such as pruning or pesticide treatments, in areas under a Mediterranean climate.

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The invasive red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera, Curculionidae), is one of the most destructive pests of palms in the world. Since its detection in the Mediterranean Basin, the ornamental Phoenix canariensis Hort. ex Chabaud has become its main host. This study was aimed at determining the life cycle of R. ferrugineus in live P. canariensis palms. Egg lethal temperature threshold and thermal constant were determined in the laboratory and resulted in 13.1°C and 40.4 degree days (DD), respectively. A semi field assay was carried out in a mesh enclosure where living P. canariensis palms were artificially infested with neonate larvae at one-month intervals from June 2008 to May 2009 under natural conditions. Infested palms were dissected at different time intervals. Maximum mortality rates for R. ferrugineus were observed for palms infested either in December or January (100%), whereas those infested from April through September showed maximum survival rates. Mean monthly temperatures below 10.3°C were lethal for neonate larvae, as 4.5°C were for older immature stages. All recovered larvae could be classed according to one of 13 instars. A thermal constant of 666.5 DD was estimated for complete larval development. Pupal develoment required an additional 282.5 DD. Based on these results and on the temperatures from 46 climatic stations selected in the Iberian Peninsula, less than one generation per year can be expected in areas with mean annual temperature below 15°C and more than two where mean annual temperature is above 19°C.

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The weevil Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) is an important pest of palms. It has recently colonized the Mediterranean Basin where it is a serious problem on ornamental Phoenix canariensis (hort. ex Chabaud) palms. The efficacy of an insecticidal paint based on chlorpyrifos and pyriproxyfen in a microencapsulated formulation (Inesfly IGR FITO, Industrias Químicas Inesba S.L., Paiporta, Spain) against this weevil has been studied. Laboratory results proved that pyriproxyfen has no effect against R. ferrugineus when applied in this microencapsulated formulation. Semifield trials dismissed Inesfly IGR FITO as a curative insecticide but showed the potential of this product in the preventative control of R. ferrugineus in palms. One single application could prevent infestation for up to 6 mo with a mean efficacy of 83.3%.

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Tetranychus urticae Koch (Acari: Tetranychidae) is an important pest of clementine mandarins, Citrus reticulata Blanco, in Spain. As a first step toward the development of an integrated crop management program for clementines, dispersion patterns of T. urticae females were determined for different types of leaves and fruit. The study was carried out between 2001 and 2003 in different commercial clementine orchards in the provinces of Castelló and Tarragona (northeastern Spain). We found that symptomatic leaves (those exhibiting typical chlorotic spots) harbored 57.1% of the total mite counts. Furthermore, these leaves were representative of mite dynamics on other leaf types. Therefore, symptomatic leaves were selected as a sampling unit. Dispersion patterns generated by Taylor's power law demonstrated the occurrence of aggregated patterns of spatial distribution (b > 1.21) on both leaves and fruit. Based on these results, the incidence (proportion of infested samples) and mean density relationship were developed. We found that optimal binomial sample sizes for estimating low populations of T. urticae on leaves (up to 0.2 female per leaf) were very large. Therefore, enumerative sampling would be more reliable within this range of T. urticae densities. However, binomial sampling was the only valid method for estimating mite density on fruit.

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Virus-like particles (VLP's) have been found in the poison glands of adult females of the parasitic wasp Opius concolor Szèpl. (Hymenoptera, Braconidae). These VLP's are found in the secretory cells either free in the cytoplasm or within cytoplasmic vesicles, sometimes associated to a secretory apparatus. Negative staining of these VLP's has revealed the occurrence of two different particles. The first type exhibits icosahedral symmetry (diameter around 70nm) and hollow surface spikes, this morphology being typical of the genus Cypovirus (Reoviridae). The other type is pleomorphic and presents an envelope with clubshaped projections (diameter ranging from 30 to 60nm), as classical textbook examples of Coronaviruses, but smaller. Function and full characterisation of these particles are not yet known.

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Laboratory toxicity trials may predict effects of chemicals under field conditions, but errors are inevitable. A chemical may be presumed harmless when in fact it has a detrimental effect, or it may appear highly toxic in the laboratory, but not in the field. Error rates depend on experimental setups, evaluation criteria, and ecological attributes, such as dietary range, of the organisms under study. The authors analyze results of standardized toxicity studies of pesticides on four species of predatory mites and assess the feasibility of drawing accurate conclusions from laboratory trials alone. This is by contrasting laboratory and field data, while varying interpretation criteria. At a 5% critical error rate, it was found that correspondence between lab and field experiments is only obtained for products harmless to Typhlodromus pyri. For this species these constitute only 30% of the total number of products in our database. Outcomes from lab tests with Amblyseius andersoni correspond with field results (for A. andersoni and A. finlandicus) either for products yielding harmless or for products yielding harmful side effects. The decision rules required to reach either classification are not compatible and hinge on field thresholds that may be unrealistic. For Phytoseiulus persimilis only harmful insecticides and harmless fungicides enabled the setting of decision rules that resulted in correspondence between lab and field trials of more than 95%. Why these species require different interpretation criteria is discussed together with suggestions for improvement of existing test protocols and the feasibility of using indicator species.

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