A strategy for mitigating avian colibacillosis disease using plant growth promoting rhizobacteria and green synthesized zinc oxide nanoparticles.

Avian colibacillosis caused by the zoonotic pathogen Escherichia coli is a common bacterial infection that causes major losses in the poultry sector. Extracts of different medicinal plants and antibiotics have been used against poultry bacterial pathogens. However, overuse of antibiotics and extracts against pathogenic strains leads to the proliferation of multi-drug resistant bacteria. Due to their environmentally friendly nature, nanotechnology and beneficial bacterial strains can be used as effective strategies against poultry infections. Green synthesis of zinc oxide nanoparticles (ZnO-NPs) from Eucalyptus globulus leaves was carried out in this study. Their characterization was done by UV-vis spectroscopy, X-ray diffraction (XRD), and Fourier transmission infrared spectroscopy (FT-IR) which confirmed their synthesis, structure, and size. In vitro, antimicrobial activities of plant leaf extract, ZnO-NPs, and plant growth-promoting rhizobacteria (PGPR) were checked against E. coli using well diffusion as well as disc diffusion method. Results proved that the antimicrobial activity of ZnO-NPs and PGPR strains was more enhanced when compared to eucalyptus leaf extract at 36 h. The maximum relative inhibition shown by ZnO-NPs, PGPR strains and eucalyptus leaf extracts was 88%, 67% and 58%, respectively. The effectiveness of ZnO-NPs was also increased with an increase in particle dose and treatment time. The 90 mg/ml of ZnO-NPs was more effective. PGPR strains from all over the tested strains, Pseudomonas sp. (HY8N) exhibited a strong antagonism against the E. coli strain as compared to other PGPR strains used in this study. However, combined application of PGPR (Pseudomonas sp. (HY8N)) and ZnO-NPs augment antagonistic effects and showed maximum 69% antagonism. The study intends to investigate the binding affinity of ZnO-NPs with the suitable receptor of the bacterial pathogen by in silico methods. The binding site conformations showed that the ligand ZnO binds with conserved binding site of penicillin-binding protein 6 (PBP 6) receptor. According to the interactions, ZnO-NPs form the same interaction pattern with respect to other reported ligands, hence it can play a significant role in the inhibition of PBP 6. This research also found that combining ZnO-NPs with Pseudomonas sp. (HY8N) was a novel and effective technique for treating pathogenic bacteria, including multidrug-resistant bacteria.

Induction of defense-related enzymes and enhanced disease resistance in maize against Fusarium verticillioides by seed treatment with Jacaranda mimosifolia formulations.

Fusarium verticillioides is an important fungal pathogen of maize, causing stalk rot and severely affecting crop production. The aim of this study was to characterize the protective effects of formulations based on Jacaranda mimosifolia leaf extracts against F. verticillioides in maize. We compared different seed treatments comprising J. mimosifolia extracts, chemical fungicide (mefenoxam) and salicylic acid to modulate the defense system of maize host plants. Both aqueous and methanolic leaf extracts of J. mimosifolia (1.2% w/v) resulted in 96-97% inhibition of mycelial growth of F. verticillioides. While a full-dose (1.2%) extract of J. mimosifolia provided significant protective effects on maize plants compared to the inoculated control, a half-dose (0.6% w/v) application of J. mimosifolia in combination with half-strength mefenoxam was the most effective treatment in reducing stalk rot disease in pot and field experiments. The same seed treatment significantly upregulated the expression of genes in the leaves encoding chitinase, glucanase, lipid transfer protein, and pathogenesis-related proteins PR-1, PR-5 and PR-10, 72 h after inoculation. This treatment also induced the activities of peroxidase, polyphenol oxidase, protease, acid invertase, chitinase and phenylalanine ammonia lyase. We conclude that seed pre-treatment with J. mimosifolia extract with half-strength chemical mefenoxam is a promising approach for the management of stalk rot in maize.

GC-MS analysis, antimicrobial, antioxidant, antilipoxygenase and cytotoxic activities of Jacaranda mimosifolia methanol leaf extracts and fractions.

Jacaranda mimosifolia trees are grown in frost-free regions globally. The aim of this study was to evaluate the methanol crude extract and various fractions of increasing polarity of J. mimosifolia leaves for bioactive metabolites, as well as antimicrobial, antioxidant and anticancer activities. The anti-inflammatory potential of the various fractions of J. mimosifolia leaf extract was studied via the lipoxygenase (LOX) inhibitory assay. Methanol crude extract (ME), derived fractions extracted with chloroform (CF) and ethyl acetate (EAF), and residual aqueous extract (AE) of dried J. mimosifolia leaves were assayed for polyphenolic compounds, their antioxidant, antimicrobial and lipoxygenase (LOX) inhibitory activities, and anticancer properties. Polyphenolic compounds were determined via HPLC while phytochemicals (total phenolics, flavonoids, tannins and ortho-diphenol contents), antioxidant activities (DPPH, hydrogen peroxideperoxide, hydroxyl and superoxide radical anions) and LOX were measured via spectrophotometry. Methanol extracts and various fractions were evaluated for antibacterial activities against Bacillus subtilis, Klebsiella pneumonia, Pseudomonas aeruginosa and Staphylococcus aureus. Antifungal potential of the fractions was tested against three species: Aspergillus flavus, Aspergillus fumigatus and Fusarium oxysporum. The highest values for total phenolic content (TPC), total flavonoid content (TFC), flavonols, tannins and ortho-diphenols were in the ME, followed by CF > EAF > AE. ME also had the highest antioxidant activity with EC50 values 48±1.3, 45±2.4, 42±1.3 and 46±1.3 µg/mL based on the DPPH, hydrogen peroxide, hydroxyl radical and superoxide radical assays, respectively. TPC and TFC showed a significant, strong and positive correlation with the values for each of these antioxidant activities. ME exhibited anti-inflammatory potential based on its LOX inhibitory activity (IC50 = 1.3 µg/mL). ME also had the maximum antibacterial and antifungal potential, followed by EAF > CF > AE. Furthermore, ME showed the strongest cytotoxic effect (EC50 = 10.7 and 17.3 µg/mL) against human hormone-dependent prostate carcinoma (LnCaP) and human lung carcinoma (LU-1) cell lines, respectively. Bioactive compounds present in leaf methanol extracts of J. mimosifolia were identified using gas chromatography-mass spectrometry (GC-MS). Fifteen compounds were identified including phenolic and alcoholic compounds, as well as fatty acids. Our results suggest that J. mimosifolia leaves are a good source of natural products with antioxidant, anti-inflammatory and anti-cancer properties for potential therapeutic, nutraceutical and functional food applications.

Nutritional value of Sesamum indicum L. was improved by Azospirillum and Azotobacter under low input of NP fertilizers.

BACKGROUND: Sesame (Sesame indicum L.) is well-known as a versatile industrial crop having various usages and contains 50-55% oil, 20% protein, 14-20% carbohydrate and 2-3% fiber. Several environmental factors are known to adversely affect yield and productivity of sesame. Our overall aim was to improve the growth, yield and quality of sesame cv. TS-3 using plant growth promoting rhizobacteria (PGPR) and saving the nitrogen and phosphate fertilizers (NP) by 50%. Field experiment (randomized complete block design) was conducted during the months of July to October of two consecutive years 2012-2013. Azospirillum (AL) and Azotobacter (AV) were applied as seed inoculation alone as well as along with half of the recommended dose of nitrogen (N) and phosphate (P) fertilizers (urea and diammonium phosphate) at the rate of 25 kg/ha and 30 kg/ha respectively. RESULTS: Here we report that A. lipoferum along with half dose of NP fertilizers (ALCF) were highly effective in increasing the agronomic and yield traits of sesame as compared to the control. A. vinelandii plus NP fertilizers (AVCF) exhibited higher seed oil content. Minimum acid value, optimum specific gravity and modified fatty acid composition were observed in ALCF treatment. Increase in oleic acid by ALCF is directly linked with improved oil quality for health benefits as oleic acid is the fatty acid which creates a balance between saturation and unsaturation of oil and for the hypotensive (blood pressure reducing) effects. CONCLUSION: It is inferred that ALCF treatment improved plant growth, seed yield and oil quality of sesame pertaining to good quality edible oil production.

Improvement of safflower oil quality for biodiesel production by integrated application of PGPR under reduced amount of NP fertilizers.

Safflower is an important industrial oil seed and bioenergy crop in semi-arid subtropical regions due to its potential to grow on marginal land and having good percentage of seed oil contents which is an important parameter for biofuel production. However, it is an ignored crop in Pakistan. In order to improve the crop productivity and reduce the use of agrochemicals for sustainable biodiesel feedstock production, an experiment was conducted for two years to improve the fatty acid composition and oil quality of Carthamus tinctorius L. (safflower) by the inoculation of Azospirillum and Azotobacter alone as well as in combined application with nitrogen and phosphate (NP) fertilizers on cultivars Thori and Saif-32 under field conditions. Separation and quantification of fatty acids were done on precise comprehensive two-dimensional gas chromatography (GC×GC). The results showed that fatty acid profile specifically monounsaturated fatty acids i-e oleic acid (C18:1) was significantly improved by Azospirillum supplemented with the quarter dose of NP fertilizers (SPQ) with concomitant decrease in polyunsaturated fatty acids by the respective treatment. Oil quality attributes such as acid value, saponification number, iodine value, refractive index and free fatty acid contents were reduced by the application of Azotobacter and Azospirillum in combination with half and quarter doses of NP fertilizers treatments (BTH, SPH, BTQ and SPQ). The reduction in these variables is positively linked with improved biodiesel yield and quality. It can be concluded that application of Azospirillum and Azotobacter not only reduced the use of NP fertilizers up to 50%-75% but also improved the oil quality in order to obtain environment friendly, sustainable and green fuel.

Botanical-chemical formulations enhanced yield and protection against Bipolaris sorokiniana in wheat by inducing the expression of pathogenesis-related proteins.

Two experiments (pot and field experiments) were conducted in two consecutive years to evaluate the protective effects of botanical-chemical formulations on physiological, biochemical performance and grain yield of wheat inoculated with Bipolaris sorokiniana. We compared different formulations comprising Calotropis procera, Jacaranda mimosifolia, Thevetia peruviana extracts, chemical fungicide (mefenoxam) and salicylic acid to modulate the defense system of wheat host plants. Among the selected plant species J. mimosifolia aqueous and methanolic leaf extracts (1.2% w/v) resulted in 96 to 97% inhibition against B. sorokiniana. Both in pot and field experiments, among all the formulations of selected plant extracts the combined formulation of JAF2 (J. mimosifolia 0.6%)+MFF2 (mefenoxam 0.1%) lowered the dose of chemical fungicide required to reduce the leaf spot blotch disease. The same formulation induced resistance in wheat apparently through the accumulation of peroxidase, polyphenol oxidase, protease, acid invertase, chitinase and phenylalanine ammonia lyase. This formulation also stimulated the defense-related gene expression of PR-proteins. The same treatment gave even more increase (48%, 12% and 22%) in no. of grains/spike, grains weight and grain yield, than the MFF1 (mefenoxam 0.2%). We conclude that foliar application of J. mimosifolia leaf extract with very low dose of chemical fungicide (J. mimosifolia 0.6%+mefenoxam 0.1%) is a promising approach for the management of leaf blight and spot blotch in wheat.

Bioinoculants: A sustainable approach to maximize the yield of Ethiopian mustard (Brassica carinata L.) under low input of chemical fertilizers.

This study aimed to find out the effect of plant growth-promoting rhizobacteria (PGPR; Azospirillum brasilense and Azotobacter vinelandii) either alone or in combination with different doses of nitrogen and phosphate fertilizers on growth, seed yield, and oil quality of Brassica carinata (L.) cv. Peela Raya. PGPR were applied as seed inoculation at 10(6) cells/mL(-1) so that the number of bacterial cells per seed was 2.6 × 10(5) cells/seed. The chemical fertilizers, namely, urea and diammonium phosphate (DAP) were applied in different doses (full dose (urea 160 kg ha(-1) + DAP 180 kg ha(-1)), half dose (urea 80 kg ha(-1) + DAP 90 kg ha(-1)), and quarter dose (urea 40 kg ha(-1) + DAP 45 kg ha(-1)). The chemical fertilizers at full and half dose significantly increased the chlorophyll, carotenoids, and protein content of leaves and the seed yield (in kilogram per hectare) but had no effect on the oil content of seed. The erucic acid (C22:1) content present in the seed was increased. Azospirillum performed better than Azotobacter and its effect was at par with full dose of chemical fertilizers (CFF) for pigments and protein content of leaves when inoculated in the presence of half dose of chemical fertilizers (SPH). The seed yield and seed size were greater. Supplementing Azospirillum with SPH assisted Azospirillum to augment the growth and yield, reduced the erucic acid (C22:1) and glucosinolates contents, and increased the unsaturation in seed oil. It is inferred that A. brasilense could be applied as an efficient bioinoculant for enhancing the growth, seed yield, and oil quality of Ethiopian mustard at low fertilizer costs and sustainable ways.

Fast comprehensive two-dimensional gas chromatography method for fatty acid methyl ester separation and quantification using dual ionic liquid columns.

Safflower oil is a complex mixture of C18 saturated and unsaturated fatty acids amongst other fatty acids, and achieving separation between these similar structure components using one dimensional gas chromatography (GC) may be difficult. This investigation aims to obtain improved separation of fatty acid methyl esters in safflower oil, and their quantification using comprehensive two-dimensional GC (GC×GC). Here, GC×GC separation is accomplished by the coupling of two ionic liquid (IL) column phases: the combination of SLB-IL111 with IL59 column phases was finally selected since it provided excellent separation of a FAME standard mixture, as well as fatty acids in safflower and linseed oil, compared to other tested column sets. Safflower oil FAME were well separated in a short run of 16min. FAME validation was demonstrated by method reproducibility, linearity over a range up to 500mgL(-1), and limits of detection which ranged from 1.9mgL(-1) to 5.2mgL(-1) at a split ratio of 20:1. Quantification was carried out using two dilution levels of 200-fold for major components and 20-fold for trace components. The fatty acids C15:0 and C17:0 were not reported previously in safflower oil. The SLB-IL111/IL59 column set proved to be an effective and novel configuration for separation and quantification of vegetable and animal oil fatty acids.