Precision and Accuracy of Field Versus Laboratory Bioassay Insecticide Efficacy for the Control of Immature Bemisia tabaci.

Ecotoxicological studies often result in reports on the limitation and sometime failures of biological assay data to predict field response to similar treatments. Nevertheless, it is widely accepted that controlled bioassays can better quantify the specific mortality response of a target pest species to a specific toxin. To quantify the relationship between whitefly bioassay and field response data, we evaluated a controlled laboratory bioassay and a concurrent cucurbit field trial method to assess insecticide efficacy for controlling the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). This was based on oviposition and nymphal development. We specifically tested the assumptions that a maximum dose bioassay would more precisely measure insecticide efficacy as compared with a comparable field spray test evaluation, and the response would be equal between the bioassay and the field as a measure of control accuracy for both adult oviposition and development of nymphal stages. To make a direct comparison, we tested the same whitefly population subsamples from 352 plots in eight cucurbit field experiments in Georgia, USA, in 2021 and 2022. The bioassays provide significantly precision for estimating proportional whitefly response. As expected, treatment-specific nonequivalence in immature whitefly counts between the bioassay and field, i.e., a lack of accuracy, only occurred with insecticides that were not highly toxic to all growth stages of whiteflies.

Determining Field Insecticide Efficacy on Whiteflies with Maximum Dose Bioassays.

We conducted a rapid bioassay method to assess insecticide efficacy for controlling adult sweetpotato whitefly Bemisia tabaci in squash and cucumber crops before insecticide applications. The study aimed to evaluate the accuracy of a 24-hour laboratory bioassay in determining maximum dose insecticide efficacy in the field. Ten insecticides were evaluated using leaf-dip bioassays, and their effectiveness was tested across eight cucurbit field experiments in Georgia, USA, during the 2021 and 2022 field seasons. The maximum dose, defined as the highest labeled rate of an insecticide diluted in the equivalent of 935 L ha-1 of water, was used for all bioassays. Adult survival observed in the bioassay was compared to adult field count-based survival 24 h after treatment. A low concentration (1/10 rate) was used for imidacloprid, flupyradifurone, pyriproxyfen, and cyantraniliprole to assess insecticide tolerance in the whitefly population. Overall, significant positive correlation between laboratory bioassay and field efficacy was reported, explaining 50-91% of the observed variation. The addition of the low dosage was helpful, indicating that no rate response was consistent with susceptibility to the tested insecticide, while a rate response was associated with a loss of susceptibility between 2021 and 2022.

Wild Radish (Raphanus raphanistrum L.) Is a Potential Reservoir Host of Cucurbit Chlorotic Yellows Virus.

Cucurbit chlorotic yellows virus (CCYV) belongs to the genus Crinivirus and is part of a complex of whitefly-transmitted viruses that cause yellowing disease in cucurbits. In the southeastern USA, heavy incidences of CCYV have been observed on all cucurbits grown in the fall. CCYV was detected from wild radish (Raphanus raphanistrum L.), a common weed that grows in the southeastern USA by high-throughput sequencing as well as RT-PCR. CCYV sequence from wild radish was 99.90% and 99.95%, identical to RNA 1 and RNA 2 of cucurbit isolates of CCYV from the region. Transmission assays using whiteflies demonstrated that wild radish is a good host for CCYV. Whiteflies were also able to acquire CCYV from wild radish and transmit the virus to cucurbit hosts, which developed typical symptoms associated with CCYV. Using quantitative PCR, the titer of CCYV in wild radish was also estimated to be on par with that of cucurbit hosts of the virus. Whitefly bioassays revealed that wild radish is an acceptable feeding and reproductive host plant. These results indicate that wild radish could serve as a reservoir host for CCYV in the USA and other parts of the world where similar conditions exist.

The effect of pyriproxyfen on the concentration of circulating metabolic fuel molecules and chemical elements in the hemolymph of Acraea horta L. (Lepidoptera: Nymphalidae): A quantitative analysis.

Many pollinating insects expand their niche to adjacent agricultural areas and are, therefore, exposed to chemical insecticides. Acraea horta L. (Lepidoptera: Nymphalidae) is a pollinator butterfly widely distributed in the Southern African region. The objectives of this work were to evaluate carbohydrate, lipid and chemical elements in the hemolymph of A. horta exposed to pyriproxyfen, a juvenile hormone analog (JHA). Last instar larvae (L6: day 1 or day 2) were topically exposed to an aqueous solution of pyriproxyfen (100 µg of the active ingredient per insect) or to diluent (control group). Hemolymph was collected after adult eclosion to determine total carbohydrate and lipid concentrations: in the control group lipids were present in lower concentrations than carbohydrates and there was no significant difference in metabolite levels between sexes; a similar pattern with similar levels were measured in the treated group, except that lipid concentrations in treated males were lower, and carbohydrate concentrations in treated females were lower than the control values. Morphologically intact adult males from treated larvae were subjected to free flight; their hemolymph carbohydrate levels were significantly reduced and did not recover to starting levels in a 30 min rest period following the exhaustive flight episode. To assess the effect of pyriproxyfen on a different stage of development, 48 h old butterflies were treated in the same way as described for the L6 larvae above; hemolymph samples were taken 48 h later for metabolite measurements and for quantification of chemical elements: carbohydrate levels decreased significantly after pyriproxyfen exposure, while lipid levels increased; inorganic elements measured in untreated adults were more abundant in females, with a general decrease in concentration following pyriproxyfen exposure, except for an increase in Fe levels in males and Cl levels in females. The quantitative changes measured in A. horta hemolymph via biochemical and chemical element analyses may indicate distinct physiological interferences beyond the main mode of action of pyriproxyfen on JH activity. In conclusion, the use and quantification of pyriproxyfen should be carefully evaluated prior to application in areas where A. horta and other pollinator species occur.

Macro-elements in the hemolymph of adult Euschistus heros (Fabr.) (Hemiptera: Pentatomidae) treated with pyriproxyfen.

Euschistus heros is an important pest in many crops in Brazil, and different control strategies, mainly involving chemicals, have been evaluated; however, the side effects of these chemicals on the balance of inorganic element levels in the hemolymph are unknown. Thus, the aim of this work was to determine the concentration of inorganic elements (focusing on macro-elements) in the hemolymph of female and male E. heros adults, after applying pyriproxyfen at a sublethal concentration (LC30 = 6.68 mL L-1 diluted in distilled water) to 4th instar nymphs, which were kept in controlled conditions. The hemolymph pool was removed 48 h after adult emergence, centrifuged and placed on an acrylic disk added with Gallium as internal standard for the analysis of total reflection X-ray fluorescence. Most of the elements in the control treatment did not differ between females and males. However, following insecticide application to females and males, respectively, there was a significant increase in sulfur (19 and 51%), chlorine (33 and 137%) and calcium (47 and 82%) in the hemolymph. The significantly higher increase in macro-elements in males' hemolymph indicates that the action of pyriproxyfen may be sex-specific. Phosphorus and potassium concentrations also differed between females and males in the control and treated groups. The observed variation in inorganic elements in the insect's hemolymph may be related to the unknown effects of pyriproxyfen, mainly on immune and reproductive performance.