Significance and interpretation of molecular diagnostics for insecticide resistance management of agricultural pests

Insecticide resistant pests become increasingly difficult to control in current day agriculture. Because of environmental and health concerns, the insecticide portfolio to combat agricultural pests is gradually decreasing. It is therefore crucial to make rational decisions on insecticide use to assure effective resistance management. However, resistance monitoring programs that inform on pest susceptibility and resistance are not yet common practice in agriculture. Molecular markers of resistance that are turned into convenient diagnostic tools are urgently needed and will only increase in importance. This review investigates which factors determine the strength, diagnostic value, and success of a diagnostic marker, and in which cases recent technical advances might provide new opportunities for decision making in an operational meaningful way

Effect of Channel Aspect Ratio on Flow Boiling Characteristics within Rectangular Micro-passages

In the present paper, a fundamental analysis on the effect of the channel aspect ratio on the bubble dynamics and heat transfer characteristics for the early transient stages of the bubble growth within confined microchannels of rectangular cross-section, under saturated flow boiling conditions, is conducted, utilising high resolution, 3D, transient, conjugate heat transfer simulations. The open-source toolbox OpenFOAM is applied for the simulations, utilising a custom, user-enhanced, diabatic Volume OF Fluid (VOF) solver. Two different series of numerical simulations are performed, focused on a single nucleation event from a single nucleation site and a single nucleation event from multiple, arbitrarily located, nucleation sites, respectively. In each series, three different values of channel aspect ratio are considered, corresponding to a narrow, a square, and a wide microchannel. For the first series, the simulations are performed for a low, a medium, and a high value of applied heat flux and mass flux. For the second series, only the lower values of applied heat flux and mass flux are used for each channel aspect ratio, since this constitutes the worst-case scenario from the overall heat transfer performance point of view, amongst the cases examined in the first series of simulations. The micro-passage aspect ratio has a significant effect in the generated bubble dynamics during the onset of the nucleate boiling regime, as the bubbles grow within the confined liquid crossflow. This alteration of the generated interfacial dynamics, in effect, regulates the size and position of the contact areas of the generated bubbles with the microchannel walls, with a direct effect in the individual contribution and therefore, the balance between the contact line and the liquid film evaporation mechanisms. Moreover, the work presents the quantification of the effect of the solid domain thermal inertia on the whole process and in particular on the local Nusselt numbers. It is evident that considering conjugate heat transfer in numerical simulations of flow boiling is compulsory in order to predict the physical processes in a correct form

The effect of hydraulic diameter on flow boiling within single rectangular microchannels and comparison of heat sink configuration of a single and multiple microchannels

Phase change heat transfer within microchannels is considered one of the most promising cooling methods for the efficient cooling of high-performance electronic devices. However, there are still fundamental parameters, such as the effect of channel hydraulic diameter Dh, whose effects on fluid flow and heat transfer characteristics are not clearly defined yet. The objective of the present work is to numerically investigate the first transient flow boiling characteristics from the bubble inception up to the first stages of the flow boiling regime development, in rectangular microchannels of varying hydraulic diameters, utilising an enhanced custom VOF-based solver. The solver accounts for conjugate heat transfer effects, implemented in OpenFOAM and validated in the literature through experimental results and analytical solutions. The numerical study was conducted through two different sets of simulations. In the first set, flow boiling characteristics in four single microchannels of Dh = 50, 100, 150, and 200 μm with constant channel aspect ratio of 0.5 and length of 2.4 mm were examined. Due to the different Dh, the applied heat and mass flux values varied between 20 to 200 kW⁄m2 and 150 to 2400 kg⁄m2s, respectively. The results of the two-phase simulations were compared with the corresponding initial single-phase stage of the simulations, and an increase of up to 37.4% on the global Nu number Nuglob was revealed. In the second set of simulations, the effectiveness of having microchannel evaporators of single versus multiple parallel microchannels was investigated by performing and comparing simulations of a single rectangular microchannel with Dh of 200 μm and four-parallel rectangular microchannels, each having a hydraulic diameter Dh of 50 μm. By comparing the local time-averaged thermal resistance along the channels, it is found that the parallel microchannels configuration resulted in a 23.3% decrease in the average thermal resistance RRl compared to the corresponding single-phase simulation stage, while the flow boiling process reduced the RRl by only 5.4% for the single microchannel case. As for the developed flow regimes, churn and slug flow dominated, whereas liquid film evaporation and, for some cases, contact line evaporation were the main contributing flow boiling mechanisms

Wettability Effect On Flow Boiling Characteristics Within Micro- passages

A numerical investigation on the effect of wettability characteristics on a single bubble growth during saturated flow boiling conditions within a microchannel, is conducted in the present paper. The numerical simulations are conducted with the open-source toolbox OpenFOAM, utilising a user-enhanced Volume OF Fluid (VOF) solver. The proposed solver enhancements involve a treatment for spurious velocities dampening (a well-known defect of VOF methods), an improved dynamic contact angle treatment to accurately account for wettability effects as well as the implementation of a phase-change model in the fluid domain, accounting for conjugate heattransfer with a solid domain. The predictions of the simulations show that the local Nusselt number (Nu) is more depended on wettability characteristics for low heat fluxes, and less dependent on higher heat fluxes. In more detail, it seems that the local, instantaneous heat transfer coefficient is higher for super-hydrophilic cases in comparison to hydrophilic. However, as the applied heat flux increases, hydrophilic and super-hydrophilic cases show a similar heat transfer enhancement with respect to the single-phase heat transfer in the considered micro-channel. Finally, superhydrophobic cases, show lower heat transfer performance with respect to the single-phase case.This is due to the fact that a vapour blanket is rapidly formed immediately after the nucleation, acting as an insulator of the heated solid surface

Numerical Investigation of Droplet Impact on Metallic Meshes

Liquid penetration analysis in porous media is of great importance in a wide range of applications such as ink jet printing technology, painting and textile design. This article presents an investigation of droplet impingement onto metallic meshes, aiming to provide insights by identifying and quantifying impact characteristics that are difficult to measure experimentally. For this purpose, an enhanced Volume-Of-Fluid (VOF) numerical simulation framework is utilised, previously developed in the general context of the OpenFOAM CFD Toolbox. Droplet impacts on metallic meshes are performed both experimentally and numerically with satisfactory degree of agreement. From the experimental investigation three main outcomes are observed—deposition, partial imbibition, and penetration. The penetration into suspended meshes leads to spectacular multiple jetting below the mesh. A higher amount of liquid penetration is linked to higher impact velocity, lower viscosity and larger pore size dimension. An estimation of the liquid penetration is given in order to evaluate the impregnation properties of the meshes. From the parametric analysis it is shown that liquid viscosity affects the adhesion characteristics of the drops significantly, whereas droplet break-up after the impact is mostly controlled by surface tension. Additionally, wettability characteristics are found to play an important role in both liquid penetration and droplet break-up below the mesh

Untangling a Gordian knot : the role of a GluCl3 I321T mutation in abamectin resistance in Tetranychus urticae

BACKGROUND: The cys-loop ligand-gated ion channels, including the glutamate-gated chloride channel (GluCl) and GABA-gated chloride channel (Rdl) are important targets for drugs and pesticides. The macrocyclic lactone abamectin primarily targets GluCl and is commonly used to control the spider mite Tetranychus urticae, an economically important crop pest. However, abamectin resistance has been reported for multiple T. urticae populations worldwide, and in several cases was associated with the mutations G314D in GluCl1 and G326E in GluCl3. Recently, an additional I321T mutation in GluCl3 was identified in several abamectin resistant T. urticae field populations. Here, we aim to functionally validate this mutation and determine its phenotypic strength. RESULTS: The GluCl3 I321T mutation was introgressed into a T. urticae susceptible background by marker-assisted backcrossing, revealing contrasting results in phenotypic strength, ranging from almost none to 50-fold. Next, we used CRISPR-Cas9 to introduce I321T, G314D and G326E in the orthologous Drosophila GluCl. Genome modified flies expressing GluCl I321T were threefold less susceptible to abamectin, while CRISPRed GluCl G314D and G326E flies were lethal. Last, functional analysis in Xenopus oocytes revealed that the I321T mutation might reduce GluCl3 sensitivity to abamectin, but also suggested that all three T. urticae Rdls are affected by abamectin. CONCLUSION: Three different techniques were used to characterize the role of I321T in GluCl3 in abamectin resistance and, combining all results, our analysis suggests that the I321T mutation has a complex role in abamectin resistance. Given the reported subtle effect, additional synergistic factors in resistance warrant more investigation

Status of Insecticide Resistance and Its Mechanisms in and Populations from Forest Settings in South Cameroon.

A key factor affecting malaria vector control efforts in Cameroon is the rapid expansion of insecticide resistance in s.l ) populations; however, mechanisms involved in insecticide resistance in forest mosquito populations are still not well documented yet. The present study was conducted to screen molecular mechanisms conferring insecticide resistance in s.l. populations from the South Cameroon forest region. WHO bioassays were conducted with F0 females aged three to four days from forest (Sangmelima, Nyabessan, and Mbandjock) and urban sites (Yaoundé (Bastos and Nkolondom)), against pyrethroids (permethrin 0.75% and deltamethrin 0.05%) and carbamates (bendiocarb 0.1%). Members of the s.l. species complex were identified using molecular diagnostic tools. TaqMan assays were used to screen for target site mutations. The expression profiles of eight genes implicated in insecticide resistance were assessed using RT-qPCR. Cuticle hydrocarbon lipids were measured to assess their potential implication in insecticide resistance. Both and were detected. was highly prevalent in Sangmelima, Nyabessan, Mbandjock, and Nkolondom. was the only species found in the Yaoundé city center (Bastos). Low mortality rate to both pyrethroids and bendiocarb was recorded in all sites. High frequency of L1014F allele (75.32-95.82%) and low frequencies of L1014S (1.71-23.05%) and N1575Y (5.28-12.87%) were recorded. The G119S mutation (14.22-35.5%) was detected for the first time in populations from Cameroon. This mutation was rather absent from populations. The detoxification genes , , , , as well as which catalyzes epicuticular hydrocarbon biosynthesis, were found to be overexpressed in at least one population. The total cuticular hydrocarvbon content, a proxy of cuticular resistance, did not show a pattern associated with pyrethroid resistance in these populations. The rapid emergence of multiple resistance mechanisms in s.l. population from the South Cameroon forest region is of big concern and could deeply affect the sustainability of insecticide-based interventions strategies in this region

Reduced proinsecticide activation by cytochrome P450 confers coumaphos resistance in the major bee parasite Varroa destructor

Varroa destructor is one of the main problems in modern beekeeping. Highly selective acaricides with low toxicity to bees are used internationally to control this mite. One of the key acaricides is the organophosphorus (OP) proinsecticide coumaphos, that becomes toxic after enzymatic activation inside Varroa. We show here that mites from the island Andros (AN-CR) exhibit high levels of coumaphos resistance. Resistance is not mediated by decreased coumaphos uptake, target-site resistance, or increased detoxification. Reduced proinsecticide activation by a cytochrome P450 enzyme was the main resistance mechanism, a powerful and rarely encountered evolutionary solution to insecticide selection pressure. After treatment with sublethal doses of [14C] coumaphos, susceptible mite extracts had substantial amounts of coroxon, the activated metabolite of coumaphos, while resistant mites had only trace amounts. This indicates a suppression of the P450 (CYP)-mediated activation step in the AN-CR mites. Bioassays with coroxon to bypass the activation step showed that resistance was dramatically reduced. There are 26 CYPs present in the V. destructor genome. Transcriptome analysis revealed overexpression in resistant mites of CYP4DP24 and underexpression of CYP3012A6 and CYP4EP4. RNA interference of CYP4EP4 in the susceptible population, to mimic underexpression seen in the resistant mites, prevented coumaphos activation and decreased coumaphos toxicity