In Silico Subtractive Proteomics and Molecular Docking Approaches for the Identification of Novel Inhibitors against Streptococcus pneumoniae Strain D39.

Streptococcus pneumoniae is a notorious Gram-positive pathogen present asymptomatically in the nasophayrnx of humans. According to the World Health Organization (W.H.O), pneumococcus causes approximately one million deaths yearly. Antibiotic resistance in S. pneumoniae is raising considerable concern around the world. There is an immediate need to address the major issues that have arisen as a result of persistent infections caused by S. pneumoniae. In the present study, subtractive proteomics was used in which the entire proteome of the pathogen consisting of 1947 proteins is effectively decreased to a finite number of possible targets. Various kinds of bioinformatics tools and software were applied for the discovery of novel inhibitors. The CD-HIT analysis revealed 1887 non-redundant sequences from the entire proteome. These non-redundant proteins were submitted to the BLASTp against the human proteome and 1423 proteins were screened as non-homologous. Further, databases of essential genes (DEGG) and J browser identified almost 171 essential proteins. Moreover, non-homologous, essential proteins were subjected in KEGG Pathway Database which shortlisted six unique proteins. In addition, the subcellular localization of these unique proteins was checked and cytoplasmic proteins were chosen for the druggability analysis, which resulted in three proteins, namely DNA binding response regulator (SPD_1085), UDP-N-acetylmuramate-L-alanine Ligase (SPD_1349) and RNA polymerase sigma factor (SPD_0958), which can act as a promising potent drug candidate to limit the toxicity caused by S. pneumoniae. The 3D structures of these proteins were predicted by Swiss Model, utilizing the homology modeling approach. Later, molecular docking by PyRx software 0.8 version was used to screen a library of phytochemicals retrieved from PubChem and ZINC databases and already approved drugs from DrugBank database against novel druggable targets to check their binding affinity with receptor proteins. The top two molecules from each receptor protein were selected based on the binding affinity, RMSD value, and the highest conformation. Finally, the absorption, distribution, metabolism, excretion, and toxicity (ADMET) analyses were carried out by utilizing the SWISS ADME and Protox tools. This research supported the discovery of cost-effective drugs against S. pneumoniae. However, more in vivo/in vitro research should be conducted on these targets to investigate their pharmacological efficacy and their function as efficient inhibitors.

Corrigendum: Effect of Allium sativum and Nigella sativa on alleviating aluminum toxicity state in the albino rats.

[This corrects the article DOI: 10.3389/fvets.2022.1042640.].

Effect of Allium sativum and Nigella sativa on alleviating aluminum toxicity state in the albino rats.

The study objective was to evaluate Allium sativum's potential and Nigella Sativa's combination's potential to reduce aluminum toxicity and return to the normal state. In the present study, a hundred albino rats were randomly divided into five equal groups. The first group was used as a control group; the other four groups were exposed to aluminum 1,600 ppm. The second exposed to aluminum only; the third and fourth groups were treated with Allium sativum 5% and Nigella sativa 5%, respectively, while the fifth group was treated with a mix of Allium sativum 2.5% and Nigella sativa 2.5% for 8th weeks. After 8 weeks, the aluminum administration was stopped, and the second group was divided into three groups. The groups were treated with Allium sativum 5% and Nigella sativa 5%, and a mix of Allium sativum 2.5% and Nigella sativa 2.5%, respectively. The first group was the control group (continued from the first experiment). Garlic and Nigella sativa were crushed and added to feed while receiving aluminum chloride daily at a dose of 1.6 ml/l was added to the drinking water. Histopathological changes in the liver, kidney, and testes were investigated after 8 and 16 weeks, and blood samples were collected after 4, 8, and 16 weeks for biochemical blood parameters. The results showed that the histopathological examination of the liver, kidney, and testes showed signs of congestion in blood vessels after aluminum exposure. Meanwhile, the treatment with Allium sativum or Nigella sativum or the mixture between them had positive effects on evading the harmful effects of aluminum in the liver, Kidney, and testes tissues. In addition, there were protective effects for Allium sativum and Nigella sativa against aluminum on serum creatinine, urea, ALT, and AST concentrations. The present study concluded that supplementation with Allium sativum or Nigella sativa or their combination could reduce aluminum toxicity and return the liver, kidney, and testes to normal.

Assessment of sublethal and transgenerational effects of spirotetramat, on population growth of cabbage aphid, Brevicoryne brassicae L. (Hemiptera: Aphididae).

The cabbage aphid (Brevicoryne brassicae L.) is a devastating pest of cruciferous crops causing economic damage worldwide and notably owing to its increasing resistance to commonly used pesticides. Such resistance prompts the development of integrated pest management (IPM) programs that include novel pesticides being effective against the aphids. Spirotetramat is a novel insecticide used against sap-sucking insect pests, particularly aphids. This study evaluated the toxicity of spirotetramat to adult apterous B. brassicae after 72 h using the leaf dipping method. According to the toxicity bioassay results, the LC50 value of spirotetramat to B. brassicae was 1.304 mgL-1. However, the sublethal concentrations (LC5 and LC15) and transgenerational effects of this novel insecticide on population growth parameters were estimated using the age-stage, two-sex life table theory method. The sublethal concentrations (LC5; 0.125 mgL-1 and LC15; 0.298 mgL-1) of spirotetramat reduced the adult longevity and fecundity of the parent generation (F0). These concentrations prolonged the preadult developmental duration while decreasing preadult survival, adult longevity and reproduction of the F1 generation. The adult pre-reproductive period was also extended by spirotetramat treatment groups. Subsequently, the population growth parameters such as the intrinsic rate of increase r, finite rate of increase λ and net reproductive rate R 0 of the F1 generation were decreased in spirotetramat treatment groups whereas, the mean generation time T of the F1 generation was not affected when compared to the control. These results indicated the negative effect of sublethal concentrations of spirotetramat on the performance of B. brassicae by reducing its nymphal survival, extending the duration of some immature stages and suppressing the population growth of B. brassicae. Overall, we demonstrated that spirotetramat is a pesticide showing both sublethal activities, and transgenerational effects on cabbage aphid; it may be useful for implementation in IPM programs against this aphid pest.

Host Resistance to Uromyces appendiculatus in Common Bean Genotypes.

Rust, induced by the fungus Uromyces appendiculatus, is one of the most serious bean diseases. The involved mechanisms in rust resistance were evaluated in 10 common bean genotypes during the 2019/2020 and 2020/2021 growing seasons. The disease parameters such as final rust severity (FRS%), area under the disease progress curve (AUDPC) and disease increase rate (r-value) were lower in the resistant genotypes than in highly susceptible genotypes. Biochemical compounds such as total phenols and the activity of antioxidant enzymes such as catalase, peroxidase and polyphenol oxidase were increased in the resistant genotypes compared to susceptible genotypes. In the resistance genotypes, the levels of oxidative stress markers such as hydrogen peroxide (H2O2) and superoxide (O2•-) increased dramatically after infection. The electrolyte leakage percentage (EL%), was found to be much greater in susceptible genotypes than resistant genotypes. The resistant gene SA14, which was found in genotypes Nebraska and Calypso at 800 bp, had an adequate level of resistance to bean rust with high grain yield potential. After infection, the transcriptions levels of 1,3-D-glucanases and phenylalanine ammonia lyase) were higher in the resistant genotypes than susceptible genotypes. In conclusion, the resistant genotypes successfully displayed desirable agronomic traits and promising expectations in breeding programs for improving management strategies of common bean rust disease. The resistance was mediated by antioxidant enzymes, phenolic compounds, and defense gene expressions, as well as the resistant gene SA14.

Mechanism of Wheat Leaf Rust Control Using Chitosan Nanoparticles and Salicylic Acid.

Wheat leaf rust is one of the world's most widespread rusts. The progress of the disease was monitored using two treatments: chitosan nanoparticles and salicylic acid (SA), as well as three application methods; spraying before or after the inoculation by 24 h, and spraying both before and after the inoculation by 24 h. Urediniospore germination was significantly different between the two treatments. Wheat plants tested for latent and incubation periods, pustule size and receptivity and infection type showed significantly reduced leaf rust when compared to untreated plants. Pucciniatriticina urediniospores showed abnormalities, collapse, lysis, and shrinkage as a result of chitosan nanoparticles treatment. The enzymes, peroxidase and catalase, were increased in the activities. In both treatments, superoxide (O2-) and hydrogen peroxide (H2O2), were apparent as purple and brown discolorations. Chitosan nanoparticles and SA treatments resulted in much more discoloration and quantitative measurements than untreated plants. In anatomical examinations, chitosan nanoparticles enhanced thickness of blade (µ), thickness of mesophyll tissue, thickness of the lower and upper epidermis and bundle length and width in the midrib compared to the control. In the control treatment's top epidermis, several sori and a large number of urediniospores were found. Most anatomical characters of flag leaves in control plants were reduced by biotic stress with P. triticina. Transcription levels of PR1-PR5 and PR10 genes were activated in chitosan nanoparticles treated plants at 0, 1 and 2 days after inoculation. In light of the data, we suggest that the prospective use of chitosan nanoparticles might be an eco-friendly strategy to improve growth and control of leaf rust disease.

Wheat Resistance to Stripe and Leaf Rusts Conferred by Introgression of Slow Rusting Resistance Genes.

Twenty-three wheat genotypes were evaluated for stripe and leaf rusts, caused by Puccinia striiformis f. sp. tritici and Puccinia triticina f. sp. tritici, respectively, at seedling and adult stages under greenhouses and field conditions during the 2019/2020 and 2020/2021 growing seasons. The race analysis revealed that 250E254 and TTTST races for stripe and leaf rusts, respectively were the most aggressive. Eight wheat genotypes (Misr-3, Misr-4, Giza-171, Gemmeiza-12, Lr34/Yr18, Lr37/Yr17, Lr46/Yr29, and Lr67/Yr46) were resistant to stripe and leaf rusts at seedling and adult stages. This result was confirmed by identifying the resistance genes: Lr34/Yr18, Lr37/Yr17, Lr46/Yr29, and Lr67/Yr46 in these genotypes showing their role in the resistance. Sids-14 and Shandweel-1 genotypes were susceptible to stripe and leaf rusts. Twelve crosses between the two new susceptible wheat genotypes and the three slow rusting genes (Lr34/Yr18, Lr37/Yr17, and Lr67/Yr46) were conducted. The frequency distribution of disease severity (%) in F2 plants of the twelve crosses was ranged from 0 to 80%. Resistant F2 plants were selected and the resistance genes were detected. This study is important for introducing new active resistance genes into the breeding programs and preserving diversity among recently released wheat genotypes.