Evidence for the Involvement of the Chemosensory Protein AgosCSP5 in Resistance to Insecticides in the Cotton Aphid, Aphis gossypii

It has been speculated that insect chemosensory proteins (CSPs) may have additional roles beyond olfaction. In this study, the phylogenetic and genomic analyses of the CSPs of the cotton aphid, Aphis gossypii, revealed the presence of gene gain-and-loss among different aphid field populations. Differential expressions of eight CSP genes were demonstrated after treatments with insecticides of different modes of action. The expression of AgosCSP5 was significantly upregulated by the insecticide treatments in a dose-dependent manner. The Drosophila flies overexpressing AgosCSP5 were significantly less susceptible to the insecticides, omethoate, imidacloprid and cypermethrin but not to deltamethrin and tau-fluvalinate, compared with control flies. The transgenic Drosophila flies exhibited an LC50 resistance ratio of 2.6 to omethoate, compared with control flies. Likewise, the mortality of the transgenic flies to imidacloprid and cypermethrin was significantly lower than that of the control flies (p < 0.01). Homology modelling, molecular docking and dynamic simulation supported the interactions and revealed a higher stability of AgosCSP5/insecticide complexes than AgosCSP5/semiochemical complexes. Our study demonstrates for first time the in vivo evidence for the involvement of CSP genes in insecticide resistance of crop insect pests and provides new insights of the newly discovered CSP-mediated insect resistance mechanism to insecticides

The chaperone balance hypothesis: The importance of the extracellular to intracellular HSP70 ratio to inflammation-driven type 2 diabetes, the effect of exercise, and the implications for clinical management

Recent evidence shows divergence between the concentrations of extracellular 70 kDa heat shock protein [eHSP70] and its intracellular concentrations [iHSP70] in people with type 2 diabetes (T2DM). A vital aspect regarding HSP70 physiology is its versatility to induce antagonistic actions, depending on the location of the protein. For example, iHSP70 exerts a powerful anti-inflammatory effect, while eHSP70 activates proinflammatory pathways. Increased eHSP70 is associated with inflammatory and oxidative stress conditions, whereas decreased iHSP70 levels are related to insulin resistance in skeletal muscle. Serum eHSP70 concentrations are positively correlated with markers of inflammation, such as C-reactive protein, monocyte count, and TNF-a, while strategies to enhance iHSP70 (e.g., heat treatment, chemical HSP70 inducers or coinducers, and physical exercise) are capable of reducing the inflammatory profile and the insulin resistance state. Here, we present recent findings suggesting that imbalances in the HSP70 status, described by the [eHSP70]/[iHSP70] ratio, may be determinant to trigger a chronic proinflammatory state that leads to insulin resistance and T2DM development. This led us to hypothesize that changes in this ratio value could be used as a biomarker for the management of the inflammatory response in insulin resistance and diabetes

Influência do uso de ractopamina na dieta de suínos em terminação sobre a concentração de lactato e cortisol sanguíneos.

Projeto: 02.06.01.001

Evolutionary trade-offs of insecticide resistance – the fitness costs associated with target-site mutations in the nAChR of Drosophila melanogaster

The evolution of resistance to drugs and pesticides poses a major threat to human health and food security. Neonicotinoids are highly effective insecticides used to control agricultural pests. They target the insect nicotinic acetylcholine receptor and mutations of the receptor that confer resistance have been slow to develop, with only one field-evolved mutation being reported to date. This is an arginine to threonine substitution at position 81 of the nAChR_β1 subunit in neonicotinoid resistant aphids. To validate the role of R81T in neonicotinoid resistance and to test whether it may confer any significant fitness costs to insects, CRISPR/Cas9 was used to introduce an analogous mutation in the genome of Drosophila melanogaster. Flies carrying R81T showed an increased tolerance (resistance) to neonicotinoid insecticides, accompanied by a significant reduction in fitness. In comparison,flies carrying a deletion of the whole nAChR_α6 subunit, the target-site of spinosyns, showed an increased tolerance to this class of insecticides but presented almost no fitness deficits

Diamide insecticide resistance in transgenic Drosophila and Sf9-cells expressing a full-length diamondback moth ryanodine receptor carrying an I4790M mutation

BACKGROUNDResistance to diamide insecticides in Lepidoptera is known to be caused primarily by amino acid changes on the ryanodine receptor (RyR). Recently, two new target site mutations, G4946V and I4790M, have emerged in populations of diamondback moth, Plutella xylostella, as well as in other lepidopteran species, and both mutations have been shown empirically to decrease diamide efficacy. Here, we quantify the impact of the I4790M mutation on diamide activation of the receptor, as compared to alterations at the G4946 locus.RESULTSI4790M when introduced into P. xylostella RyR expressed in an insect-derived Sf9 cell line was found to mediate just a ten-fold reduction in chlorantraniliprole efficacy (compared to 104- and 146-fold reductions for the G4946E and G4946V variants, respectively), whilst in the field its presence is associated with a ≥150-fold reduction. I4790M-mediated resistance to flubendiamide was estimated to be >24-fold. When the entire coding sequence of P. xylostella RyR was integrated into Drosophila melanogaster, the I4790M variant conferred ~4.4-fold resistance to chlorantraniliprole and 22-fold resistance to flubendiamide in the 3rd instar larvae, confirming that it imparts only a moderate level of resistance to diamide insecticides. Although the I4790M substitution appears to bear no fitness costs in terms of the flies' reproductive capacity, when assessed in a noncompetitive environment, it does, however, have potentially major impacts on mobility at both the larval and adult stages.CONCLUSIONSI4790M imparts only a moderate level of resistance to diamide insecticides and potentially confers significant fitness costs to the insect

Legume and Nitrogen Fertilization Affect Animal Performance and Enteric Methane Emission of Nellore Heifers

Methane emission from livestock operation is an important source of greenhouse gas and contributes to global warming. Forage legume secondary compounds may mitigate methane emissions by reducing methanogenic population in the rumen. This study evaluated animal performance and methane emission from beef cattle grazing either a mixed pasture [Brachiaria brizantha cv. Marandu (palisadegrass) and Arachis pintoi (forage peanut) cv. BRS Mandobi] or a palisadegrass monoculture with or without nitrogen (N) fertilisation. A 2.5-yr continuous stocking experiment was carried out in southeast Brazil, on a randomized complete block design with three treatments and four replicates. Two Nellore heifers were used as tester animals and additional put-and-takes were used to keep canopy height at 20-25 cm. The treatments comprised three pasture types: 1) palisadegrass-forage peanut mixed pasture (GRASS+LEGUME); 2) palisadegrass + 150 kg N/ha/year (GRASS+N); 3) palisadegrass without N fertilization (GRASS). Response variables included average daily gain (ADG), forage intake, and methane emission. Methane emission was estimated by the sulphur hexafluoride (SF6) tracer technique. There was no difference between grazing systems for the ADG (P = 0.439) and DMI (P = 0.394; averages of 0.433 kg/d and 2.10 %BW/d, respectively). In the GRASS+LEGUME, there was a decrease of 11.7% in methane emission per animal (148 vs. 170 and 165 g/day for GRASS+N and GRASS, respectively; P = 0.001). Grazing systems including legume reduced methane emission per unit of ADG (365 vs. 428 and 398 g/kg for GRASS and GRASS+N, respectively; P = 0.061) and per carcass gain (656 vs. 800 g of methane/kg carcass for GRASS; P = 0.022). Intake of condensed tannins was greater for GRASS+LEGUME (0.61 vs. 0.17 %BW/d, P \u3c 0.001). Forage peanut decreased enteric methane emission intensity, reducing carbon footprint of livestock systems in Southeast Brazil

Physical, thermal, and antibacterial effects of active essential oils with potential for biomedical applications loaded onto cellulose acetate/polycaprolactone wet-spun microfibers

New approaches to deal with the growing concern associated with antibiotic-resistant bacteria are emerging daily. Essential oils (EOs) are natural antimicrobial substances with great potential to mitigate this situation. However, their volatile nature, in their liquid-free form, has restricted their generalized application in biomedicine. Here, we propose the use of cellulose acetate (CA)/polycaprolactone (PCL) wet-spun fibers as potential delivery platforms of selected EOs to fight infections caused by Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). Twenty EOs were selected and screened for their minimal inhibitory concentration (MIC), using the antibiotic ampicillin as positive control. The cinnamon leaf oil (CLO), cajeput oil (CJO), and the clove oil (CO) were the most effective EOs, against the Gram-positive (MIC < 22.38 mg/mL) and the Gram-negative (MIC < 11.19 mg/mL) bacteria. Uniform microfibers were successfully wet-spun from CA/PCL with an averaged diameter of 53.9 ± 4.5 µm, and then modified by immersion with CLO, CJO and CO at 2 × MIC value. EOs incorporation was confirmed by UV-visible spectroscopy, Fourier-transformed infrared spectroscopy, and thermal gravimetric analysis. However, while microfibers contained ampicillin at MIC (control) after the 72 h modification, the CLO, CO and CJO-loaded fibers registered ≈ 14%, 66%, and 76% of their MIC value, respectively. Data showed that even at small amounts the EO-modified microfibers were effective against the tested bacteria, both by killing bacteria more quickly or by disrupting more easily their cytoplasmic membrane than ampicillin. Considering the amount immobilized, CLO-modified fibers were deemed the most effective from the EOs group. These results indicate that CA/PCL microfibers loaded with EOs can be easily produced with increased antibacterial action, envisioning their use as scaffolding materials for the treatment of infections.Authors acknowledge the Portuguese Foundation for Science and Technology (FCT), FEDER funds by means of Portugal 2020 Competitive Factors Operational Program (POCI) and the Portuguese Government(OE) for funding the project PEPTEX with reference PTDC/CTM-TEX/28074/2017 (POCI-01-0145-FEDER-028074).Authors also acknowledge project UID/CTM/00264/2020 of Centre for Textile Science and Technology (2C2T),funded by national funds through FCT/MCTES. SEM studies were performed at the Materials CharacterizationServices of the University of Minho (SEMAT/UM)

Biodegradable wet-spun fibers modified with antimicrobial agents for potential applications in biomedical engineering

Wet-spinning is a non-solvent induced phase inversion technique that allows the production of continuous polymeric microfibers, with a uniform morphology, based on the principle of precipitation. It allows the production of 3D fibrous constructs with an intricated architecture that facilitates cell infiltration, something that is very limited in electrospun nanofibrous mats, thus increasing its interest in biomedicine. Wet-spun scaffolds are also more easily processed and can be loaded with a variety of biomolecules of interest. Antimicrobial agents that display a broad spectrum of activity against bacteria, fungi and viruses have been combined with such constructs demonstrating great potential to fight infections. In the present work, we explore the use of wet-spinning to process both natural and synthetic biodegradable polymers in the form of microfibers, and the necessary processes to modify their surface to increase their antimicrobial profile. The synergistic potential of specialized biomolecules within wet-spun fibrous architectures are also highlighted.Authors acknowledge the Foundation for Science and Technology (FCT) of Portugal for funding the projects PTDC/CTM-TEX/28074/2017 (POCI-01-0145-FEDER-028074) and UID/CTM/00264/2020 from 2C2T