scholarly journals Targeting a coatomer protein complex-I gene via RNA interference results in effective lethality in the pollen beetle Brassicogethes aeneus

2020 ◽  
Author(s):  
Jonathan Willow ◽  
Silva Sulg ◽  
Clauvis Nji Tizi Taning ◽  
Ana Isabel Silva ◽  
Olivier Christiaens ◽  
...  
Keyword(s):  
Rna Interference ◽  
Control Strategies ◽  
Dietary Exposure ◽  
Pyrethroid Insecticides ◽  
Pollen Beetle ◽  
Nontarget Organisms ◽  
Brassicogethes Aeneus ◽  
Development Of Resistance ◽  
Species Specific ◽  
The Impact

AbstractThe pollen beetle Brassicogethes aeneus is a serious pest of oilseed rape (Brassica napus) in Europe. Management of this pest has grown difficult due to B. aeneus’s development of resistance to pyrethroid insecticides, as well as the pressure to establish control strategies that minimise the impact on nontarget organisms. RNA interference represents a nucleotide sequence-based, and thus potentially species-specific, approach to agricultural pest control. The present study examined the efficacy of targeting the coatomer gene coatomer subunit alpha (αCOP), via both microinjection and dietary exposure to exogenous complementary dsRNA, on αCOP-silencing and subsequent mortality in B. aeneus. Beetles injected with dsRNA targeting αCOP (at 0.14 µg/mg) showed 88% and 100% mortality at 6 and 10 days post-injection, respectively; where by the same time after dietary exposure, 43%–89% mortality was observed in the 3 µg dsRNA/µL treatment, though the effect was concentration-dependent. Thus, the effect was significant for both delivery routes. In working towards RNA-based management of B. aeneus, future studies should include αCOP as a target of interest.

scholarly journals RNAi efficacy is enhanced by chronic dsRNA feeding in pollen beetle

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Jonathan Willow ◽  
Liina Soonvald ◽  
Silva Sulg ◽  
Riina Kaasik ◽  
Ana Isabel Silva ◽  
...  
Keyword(s):  
Rna Interference ◽  
Pest Management ◽  
Oilseed Rape ◽  
Growth Stages ◽  
Short Term ◽  
Pollen Beetle ◽  
Brassicogethes Aeneus ◽  
Crop Growth Stages ◽  
Insecticidal Compounds ◽  
Significant Mortality

AbstractDouble-stranded RNAs (dsRNAs) represent a promising class of biosafe insecticidal compounds. We examined the ability to induce RNA interference (RNAi) in the pollen beetle Brassicogethes aeneus via anther feeding, and compared short-term (3 d) to chronic (17 d) feeding of various concentrations of dsRNA targeting αCOP (dsαCOP). In short-term dsαCOP feeding, only the highest concentration resulted in significant reductions in B. aeneus survival; whereas in chronic dsαCOP feeding, all three concentrations resulted in significant mortality. Chronic dsαCOP feeding also resulted in significantly greater mortality compared to short-term feeding of equivalent dsαCOP concentrations. Our results have implications for the economics and development of dsRNA spray approaches for managing crop pests, in that multiple lower-concentration dsRNA spray treatments across crop growth stages may result in greater pest management efficacy, compared to single treatments using higher dsRNA concentrations. Furthermore, our results highlight the need for research into the development of RNAi cultivars for oilseed rape protection, given the enhanced RNAi efficacy resulting from chronic, compared to short-term, dsRNA feeding in B. aeneus.

scholarly journals First Evidence of Bud Feeding-Induced RNAi in a Crop Pest via Exogenous Application of dsRNA

Insects ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 769
Author(s):  
Jonathan Willow ◽  
Liina Soonvald ◽  
Silva Sulg ◽  
Riina Kaasik ◽  
Ana Isabel Silva ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Gene Silencing ◽  
Oilseed Rape ◽  
Dietary Exposure ◽  
Exposure Period ◽  
Sequence Specificity ◽  
Pollen Beetle ◽  
Brassicogethes Aeneus ◽  
Model Control ◽  
Potential Strategy

Spray-induced gene silencing (SIGS) is a potential strategy for agricultural pest management, whereby nucleotide sequence-specific double-stranded RNA (dsRNA) can be sprayed onto a crop; the desired effect being a consumption of dsRNA by the target pest, and subsequent gene silencing-induced mortality. Nucleotide sequence-specificity is the basis for dsRNA’s perceived biosafety. A biosafe approach to pollen beetle (Brassicogethes aeneus) management in oilseed rape (Brassica napus) agroecosystems is needed. We examined the potential for SIGS in B. aeneus, via bud feeding, a field-relevant dsRNA exposure route. Oilseed rape buds were uniformly treated with dsRNA designed to target αCOP in B. aeneus. Our model control dsRNA (dsGFP) remained detectable on buds throughout the entire 3 d exposure period. When applied at 5 µg/µL, dsαCOP induced significant αCOP silencing 3 d after dietary exposure to buds treated with this dsαCOP concentration. We also observed a trend of increased αCOP silencing with increasing concentrations of dsαCOP at both 3 and 6 d. Furthermore, we observed a marginally significant and significant reduction in B. aeneus survival at 10 and 15 d, respectively. Our results suggest potential for developing a SIGS approach to B. aeneus management—though further experiments are needed to more fully understand this potential.

scholarly journals The effect of insecticide application by dropleg sprayers on pollen beetle parasitism in oilseed rape

BioControl ◽  
2021 ◽  
Author(s):  
Johannes Hausmann ◽  
Udo Heimbach ◽  
Michael Rostás ◽  
Meike Brandes
Keyword(s):  
Oilseed Rape ◽  
Insect Pests ◽  
Biological Control Agent ◽  
Control Agent ◽  
Total Catch ◽  
Pollen Beetle ◽  
Brassicogethes Aeneus ◽  
Larval Parasitoid ◽  
Parasitism Rates ◽  
The Impact

AbstractDropleg sprayers apply pesticides below the flower horizon of oilseed rape plants and thus reduce unwanted side effects on pollinating insects. Whether this technique benefits parasitoids of seed and pollen feeding insect pests has not been studied earlier. To answer this question, we first assessed the vertical distribution of pests and parasitoids using a portable aspirator. In addition, parasitism rates of pollen beetle, Brassicogethes aeneus Fabricius (Coleoptera: Nitidulidae), by the larval parasitoid Tersilochus heterocerus Thomson (Hymenoptera: Ichneumonidae) were compared in conventional and dropleg sprayed fields over four years (2016–2019), using the neonicotinoids thiacloprid and acetamiprid. Our results show that seed and pollen feeders were mainly found in the flowering canopy, while the predominant location of parasitoids was species-specific. Among pollen beetle parasitoids, Phradis interstitialis Thomson (Hymenoptera: Ichneumonidae) was more abundant below flowering canopy (63% of total catch), whereas T. heterocerus was mainly caught in the flowering canopy (84% of total catch). In the spraying experiments, average parasitism rates of pollen beetles by T. heterocerus ranged between 55 and 82% in the untreated controls. In the dropleg spray treatments, parasitism rates did not differ significantly from control levels, with the exception of thiacloprid application in 2019. In contrast, conventional spray applications resulted in a reduction of parasitism rates by up to 37% compared to the control for at least one of the insecticides in three out of four years. The impact of conventional application differed between years, which may be explained by the temporal coincidence between spray application and the immigration of parasitoids into the crop. We conclude that dropleg spraying exerts lower non-target effects on the main biological control agent of pollen beetle.

A curated collection of Klebsiella metabolic models reveals variable substrate usage and gene essentiality

2021 ◽  
Author(s):  
Jane Hawkey ◽  
Ben Vezina ◽  
Jonathan Monk ◽  
Louise M Judd ◽  
Taylor Harshegyi ◽  
...  
Keyword(s):  
Control Strategies ◽  
Single Gene ◽  
Multi Drug Resistance ◽  
Growth Data ◽  
Gene Deletions ◽  
Diverse Range ◽  
Metabolic Models ◽  
Species Specific ◽  
Healthcare Associated ◽  
The Impact

The Klebsiella pneumoniae species complex (KpSC) is a set of seven Klebsiella taxa which are found in a variety of niches, and are an important cause of opportunistic healthcare- associated infections in humans. Due to increasing rates of multi-drug resistance within the KpSC, there is a growing interest in better understanding the biology and metabolism of these organisms to inform novel control strategies. We collated 37 sequenced KpSC isolates isolated from a variety of niches, representing all seven taxa. We generated strain-specific genome scale metabolic models (GEMs) for all 37 isolates and simulated growth phenotypes on 511 distinct carbon, nitrogen, sulphur and phosphorus substrates. Models were curated and their accuracy assessed using matched phenotypic growth data for 94 substrates (median accuracy of 96%). We explored species-specific growth capabilities and examined the impact of all possible single gene deletions on growth in 145 core carbon substrates. These analyses revealed multiple strain-specific differences, within and between species and highlight the importance of selecting a diverse range of strains when exploring KpSC metabolism. This diverse set of highly accurate GEMs could be used to inform novel drug design, enhance genomic analyses, and identify novel virulence and resistance determinants. We envisage that these 37 curated strain-specific GEMs, covering all seven taxa of the KpSC, provide a valuable resource to the Klebsiella research community.

scholarly journals Power Hardware-in-the-Loop-Based Performance Analysis of Different Converter Controllers for Fast Active Power Regulation in Low-Inertia Power Systems

Energies ◽  
2021 ◽  
Vol 14 (11) ◽  
pp. 3274
Author(s):  
Jose Rueda Torres ◽  
Zameer Ahmad ◽  
Nidarshan Veera Kumar ◽  
Elyas Rakhshani ◽  
Ebrahim Adabi ◽  
...  
Keyword(s):  
Performance Evaluation ◽  
Power Systems ◽  
Real Time ◽  
Control Strategies ◽  
Active Power ◽  
Power Electronic ◽  
Hardware In The Loop ◽  
Digital Controllers ◽  
Power Regulation ◽  
The Impact

Future electrical power systems will be dominated by power electronic converters, which are deployed for the integration of renewable power plants, responsive demand, and different types of storage systems. The stability of such systems will strongly depend on the control strategies attached to the converters. In this context, laboratory-scale setups are becoming the key tools for prototyping and evaluating the performance and robustness of different converter technologies and control strategies. The performance evaluation of control strategies for dynamic frequency support using fast active power regulation (FAPR) requires the urgent development of a suitable power hardware-in-the-loop (PHIL) setup. In this paper, the most prominent emerging types of FAPR are selected and studied: droop-based FAPR, droop derivative-based FAPR, and virtual synchronous power (VSP)-based FAPR. A novel setup for PHIL-based performance evaluation of these strategies is proposed. The setup combines the advanced modeling and simulation functions of a real-time digital simulation platform (RTDS), an external programmable unit to implement the studied FAPR control strategies as digital controllers, and actual hardware. The hardware setup consists of a grid emulator to recreate the dynamic response as seen from the interface bus of the grid side converter of a power electronic-interfaced device (e.g., type-IV wind turbines), and a mockup voltage source converter (VSC, i.e., a device under test (DUT)). The DUT is virtually interfaced to one high-voltage bus of the electromagnetic transient (EMT) representation of a variant of the IEEE 9 bus test system, which has been modified to consider an operating condition with 52% of the total supply provided by wind power generation. The selected and programmed FAPR strategies are applied to the DUT, with the ultimate goal of ascertaining its feasibility and effectiveness with respect to the pure software-based EMT representation performed in real time. Particularly, the time-varying response of the active power injection by each FAPR control strategy and the impact on the instantaneous frequency excursions occurring in the frequency containment periods are analyzed. The performed tests show the degree of improvements on both the rate-of-change-of-frequency (RoCoF) and the maximum frequency excursion (e.g., nadir).

scholarly journals Estimating asymptomatic, undetected and total cases for the COVID-19 outbreak in Wuhan: a mathematical modeling study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xi Huo ◽  
Jing Chen ◽  
Shigui Ruan
Keyword(s):  
Disease Transmission ◽  
Large Scale ◽  
Transmission Rate ◽  
Control Strategies ◽  
Herd Immunity ◽  
Antibody Prevalence ◽  
Screening Process ◽  
Vaccination Programs ◽  
Prevalence Level ◽  
The Impact

Abstract Background The COVID-19 outbreak in Wuhan started in December 2019 and was under control by the end of March 2020 with a total of 50,006 confirmed cases by the implementation of a series of nonpharmaceutical interventions (NPIs) including unprecedented lockdown of the city. This study analyzes the complete outbreak data from Wuhan, assesses the impact of these public health interventions, and estimates the asymptomatic, undetected and total cases for the COVID-19 outbreak in Wuhan. Methods By taking different stages of the outbreak into account, we developed a time-dependent compartmental model to describe the dynamics of disease transmission and case detection and reporting. Model coefficients were parameterized by using the reported cases and following key events and escalated control strategies. Then the model was used to calibrate the complete outbreak data by using the Monte Carlo Markov Chain (MCMC) method. Finally we used the model to estimate asymptomatic and undetected cases and approximate the overall antibody prevalence level. Results We found that the transmission rate between Jan 24 and Feb 1, 2020, was twice as large as that before the lockdown on Jan 23 and 67.6% (95% CI [0.584,0.759]) of detectable infections occurred during this period. Based on the reported estimates that around 20% of infections were asymptomatic and their transmission ability was about 70% of symptomatic ones, we estimated that there were about 14,448 asymptomatic and undetected cases (95% CI [12,364,23,254]), which yields an estimate of a total of 64,454 infected cases (95% CI [62,370,73,260]), and the overall antibody prevalence level in the population of Wuhan was 0.745% (95% CI [0.693%,0.814%]) by March 31, 2020. Conclusions We conclude that the control of the COVID-19 outbreak in Wuhan was achieved via the enforcement of a combination of multiple NPIs: the lockdown on Jan 23, the stay-at-home order on Feb 2, the massive isolation of all symptomatic individuals via newly constructed special shelter hospitals on Feb 6, and the large scale screening process on Feb 18. Our results indicate that the population in Wuhan is far away from establishing herd immunity and provide insights for other affected countries and regions in designing control strategies and planing vaccination programs.

scholarly journals 152 Impact of chorionic somatomammotropin RNA interference on uterine blood flow and placental glucose uptake in the absence of intrauterine growth restriction

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 121-121
Author(s):  
Amelia R Tanner ◽  
Asghar Ali ◽  
Quinton A Winger ◽  
Paul J Rozance ◽  
Russell V Anthony
Keyword(s):  
Rna Interference ◽  
Glucose Uptake ◽  
Intrauterine Growth Restriction ◽  
Growth Restriction ◽  
Intrauterine Growth ◽  
Blood Flows ◽  
Umbilical Blood ◽  
Exact Function ◽  
Chorionic Somatomammotropin ◽  
The Impact

Abstract Chorionic somatomammotropin (CSH) is one of the most abundant hormones produced by the sheep placenta, yet the exact function of CSH has been elusive. Previously we reported the use of in vivo RNA interference (RNAi) to assess the impact of CSH deficiency on placental and fetal growth in sheep. Near-term (135 dGA), there are two distinct CSH RNAi phenotypes: 1) pregnancies with intrauterine growth restriction (IUGR), and 2) pregnancies with normal fetal and placental weights. This study describes physiological changes in the latter phenotype. To generate the CSH RNAi pregnancies, the trophectoderm of hatched blastocysts (9 dGA) were infected with lentiviral-constructs expressing either a scrambled control (NTS) or CSH-specific shRNA (CSH RNAi), prior to transfer into synchronized recipient ewes. At 120 dGA, 6 NTS and 6 CSH RNAi pregnancies were fitted with maternal and fetal catheters. Uterine and umbilical blood flows were measured utilizing the 3H2O transplacental diffusion technique at 132 dGA, and nutrient uptakes were calculated by the Fick principle. Resulting data were analyzed by Student’s t-test and significance was set at P ≤ 0.05. CSH RNAi tended (P ≤ 0.10) to reduce placentome weight with no effect on fetal weight. Absolute (ml/min) and relative (ml/min/kg fetus) uterine blood flows were reduced (P ≤ 0.05) in CSH RNAi pregnancies, but umbilical flows were not impacted. The uterine artery-to-vein glucose gradient (mmol/l) was significantly (P ≤ 0.05) increased, whereas the gradients for taurine and glycine were reduced (P ≤ 0.05). Uteroplacental glucose uptake (mmol/min/kg placenta) was increased 27% (P ≤ 0.05), whereas umbilical glucose uptake (mmol/min/kg fetus) was reduced 13%. This cohort demonstrates that even in the absence of IUGR, CSH deficiency has significant physiological ramifications, and the investigation of CSH RNAi pregnancies exhibiting both IUGR and non-IUGR phenotypes may help determine the direct effects of CSH and its potential impact on fetal programming. Supported by NIH R01 HD093701.

scholarly journals Codon harmonization reduces amino acid misincorporation in bacterially expressed P. falciparum proteins and improves their immunogenicity

AMB Express ◽  
2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Neeraja Punde ◽  
Jennifer Kooken ◽  
Dagmar Leary ◽  
Patricia M. Legler ◽  
Evelina Angov
Keyword(s):  
Protein Structure ◽  
Amino Acid ◽  
Codon Usage ◽  
Dna Sequences ◽  
Structural Integrity ◽  
Host Cells ◽  
Loss Of Function ◽  
Species Specific ◽  
And Function ◽  
The Impact

Abstract Codon usage frequency influences protein structure and function. The frequency with which codons are used potentially impacts primary, secondary and tertiary protein structure. Poor expression, loss of function, insolubility, or truncation can result from species-specific differences in codon usage. “Codon harmonization” more closely aligns native codon usage frequencies with those of the expression host particularly within putative inter-domain segments where slower rates of translation may play a role in protein folding. Heterologous expression of Plasmodium falciparum genes in Escherichia coli has been a challenge due to their AT-rich codon bias and the highly repetitive DNA sequences. Here, codon harmonization was applied to the malarial antigen, CelTOS (Cell-traversal protein for ookinetes and sporozoites). CelTOS is a highly conserved P. falciparum protein involved in cellular traversal through mosquito and vertebrate host cells. It reversibly refolds after thermal denaturation making it a desirable malarial vaccine candidate. Protein expressed in E. coli from a codon harmonized sequence of P. falciparum CelTOS (CH-PfCelTOS) was compared with protein expressed from the native codon sequence (N-PfCelTOS) to assess the impact of codon usage on protein expression levels, solubility, yield, stability, structural integrity, recognition with CelTOS-specific mAbs and immunogenicity in mice. While the translated proteins were expected to be identical, the translated products produced from the codon-harmonized sequence differed in helical content and showed a smaller distribution of polypeptides in mass spectra indicating lower heterogeneity of the codon harmonized version and fewer amino acid misincorporations. Substitutions of hydrophobic-to-hydrophobic amino acid were observed more commonly than any other. CH-PfCelTOS induced significantly higher antibody levels compared with N-PfCelTOS; however, no significant differences in either IFN-γ or IL-4 cellular responses were detected between the two antigens.

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