Protecting honey bees: identification of a new varroacide by in silico, in vitro, and in vivo studies.

Varroa destructor is the main concern related to the gradual decline of honeybees. Nowadays, among the various acaricides used in the control of V. destructor, most presents increasing resistance. An interesting alternative could be the identification of existent molecules as new acaricides with no effect on honeybee health. We have previously constructed the first 3D model of AChE for honeybee. By analyzing data concerning amino acid mutations implicated in the resistance associated to pesticides, it appears that pirimicarb should be a good candidate for varroacide. To check this hypothesis, we characterized the AChE gene of V. destructor. In the same way, we proposed a 3D model for the AChE of V. destructor. Starting from the definition of these two 3D models of AChE in honeybee and varroa, a comparison between the gorges of the active site highlighted some major differences and particularly different shapes. Following this result, docking studies have shown that pirimicarb adopts two distinct positions with the strongest intermolecular interactions with VdAChE. This result was confirmed with in vitro and in vivo data for which a clear inhibition of VdAChE by pirimicarb at 10 µM (contrary to HbAChE) and a 100% mortality of varroa (dose corresponding to the LD50 (contact) for honeybee divided by a factor 100) were observed. These results demonstrate that primicarb could be a new varroacide candidate and reinforce the high relationships between in silico, in vitro, and in vivo data for the design of new selective pesticides.

Suitability of two laboratory testing methods to evaluate the side effects of pesticides on Typhlodromus pyri Scheuten (Acari: Phytoseiidae).

BACKGROUND: Laboratory results of the French ANPP/CEB guideline No. 167 and IOBC/WPRS Ring Testing Group methods for testing the side effects of pesticides on the predatory mite Typhlodromus pyri Scheuten were compared with respect to their suitability to evaluate the toxicity of three pesticides. RESULTS: Results obtained with the ANPP/CEB guideline allow the demonstration of significant differences between two slightly toxic products, a dichlofluanid 500 g kg(-1) kWP (Euparen) 50WP) and a quinoxyfen 250 g L(-1) SCC (Legend), and a highly toxic cymoxanil 60/mancozeb 200/folpet 275 g kg(-1) WP [Remiltine F Pepite) (RFP)], on the basis of bioassays conducted in the laboratory. In contrast, results obtained with the IOBC/WPRS method classified all three as harmful. CONCLUSION: The evaluation of the toxicity of RFP revealed that the concentration, the quantity of the wet deposit and the food source used in the IOBC/WPRS method maximise the toxicity, in comparison with those used in the ANPP/CEB protocol. Valid criteria in controls were all respected in the ANPP/CEB tests but not in the IOBC/WPRS samples. This result revealed difficulties related to the use of the IOBC/WPRS method in laboratories.

Repellent, irritant and toxic effects of 20 plant extracts on adults of the malaria vector Anopheles gambiae mosquito.

Pyrethroid insecticides induce an excito-repellent effect that reduces contact between humans and mosquitoes. Insecticide use is expected to lower the risk of pathogen transmission, particularly when impregnated on long-lasting treated bednets. When applied at low doses, pyrethroids have a toxic effect, however the development of pyrethroid resistance in several mosquito species may jeopardize these beneficial effects. The need to find additional compounds, either to kill disease-carrying mosquitoes or to prevent mosquito contact with humans, therefore arises. In laboratory conditions, the effects (i.e., repellent, irritant and toxic) of 20 plant extracts, mainly essential oils, were assessed on adults of Anopheles gambiae, a primary vector of malaria. Their effects were compared to those of DEET and permethrin, used as positive controls. Most plant extracts had irritant, repellent and/or toxic effects on An. gambiae adults. The most promising extracts, i.e. those combining the three types of effects, were from Cymbopogon winterianus, Cinnamomum zeylanicum and Thymus vulgaris. The irritant, repellent and toxic effects occurred apparently independently of each other, and the behavioural response of adult An. gambiae was significantly influenced by the concentration of the plant extracts. Mechanisms underlying repellency might, therefore, differ from those underlying irritancy and toxicity. The utility of the efficient plant extracts for vector control as an alternative to pyrethroids may thus be envisaged.

Immigration of phytoseiid mites from surrounding uncultivated areas into a newly planted vineyard.

This study reports (1) a faunistic survey of phytoseiid mites observed inside a vine plot and in neighbouring vegetation (other vine plots and uncultivated areas) and (2) dispersal of phytoseiid mites into the plot studied. These data aim to raise some hypotheses concerning natural colonisation of a vineyard by predatory mites. The study was carried out over 3 years (1999, 2000 and 2001) in an experimental plot planted with two cultivars (Grenache and Syrah) and with Sorbus domestica in a framework of agroforestry investigations. Phytoseiid mites were collected in both cultivated and uncultivated areas surrounding the experimental plot, and their dispersal into the plot studied using "aerial" traps. Densities remained quite low compared to previous studies. The main species encountered in the uncultivated areas and in the traps was Typhlodromus phialatus. Despite the low numbers of phytoseiid mites trapped, densities of phytoseiid mites into the vine field increased during 3 years. Typhlodromus phialatus, the species mainly found in the neighbouring uncultivated areas, was rarely found in vineyards. Another morphologically close species was predominant on vines: Typhlodromus exhilaratus. However, Kampimodromus aberrans the main phytoseiid mite species in vineyards of Southern France was not found in the present survey. Hypotheses for this colonisation process are discussed.

A genetic analysis of mancozeb resistance in Typhlodromus pyri (Acari: Phytoseiidae).

A field population of Typhlodromus pyri (Acari: Phytoseiidae) tolerant to mancozeb was selected in the laboratory. After 10 mancozeb selections the LC50 value for mancozeb was 73 times higher in the selected-10 strain compared to the standard susceptible strain. A genetic analysis using reciprocal crosses and backcrosses of female F1 progeny found no maternal effect. Resistance in the selected-10 strain was codominant in expression, dominance value was about -0.1. Backcrosses between F1 females and the susceptible strain indicate that the resistance to mancozeb could be principally conferred by a predominant gene, but additional factors would also be involved.