Microbial-based metabolites associated with degradation of imidacloprid and its impact on stress-responsive proteins.

Imidacloprid (IMD), a neonicotinoid insecticide, is intensively used in agricultural fields for effective protection against aphids, cane beetles, thrips, stink bugs, locusts, etc., is causing serious environmental concerns. In recent years, seed treatment with Imidacloprid is being practiced mainly to prevent sucking insect pests. In India, due to the increase in application of this insecticide residue has been proven to have an impact on the quality of soil and water. In view of this, the current investigation is focussed on sustainable approach to minimize the residual effect of IMD in agricultural fields. The present study reveals a most promising imidacloprid resistant bacterium Lysinibacillus fusiformis IMD-Bio5 strain isolated from insecticide-contaminated soil. The isolated bacterial strain upon tested for its biodegradation potential on mineral salt medium (MSM) showed a significant survival growth at 150 g/L of IMD achieved after 3 days, whereas immobilized cells on MSM amended with 200 g/L of IMD as the sole carbon source provided degradation of 188 and 180 g/L of IMD in silica beads and sponge matrices, respectively. The liquid chromatography mass spectrometry was performed to test the metabolite responsive for IMD biodegradation potential of L. fusiformis IMD-Bio5 which showed the induced activity of the metabolite 6-Chloronicotinic acid. Furthermore, as compared to the untreated control, the Lysinibacillus fusiformis IMD-Bio5 protein profile revealed a range of patterns showing the expression of stress enzymes. Thus, results provided a most effective bacterium enabling the removal of IMD-like hazardous contaminants from the environment, which contributes to better agricultural production and soil quality, while long-term environmental advantages are restored.

Bacterial chemotaxis of herbicide atrazine provides an insight into the degradation mechanism through intermediates hydroxyatrazine, N-N-isopropylammelide, and cyanuric acid compounds.

In recent times, the herbicide atrazine (ATZ) has been commonly used before and after the cultivation of crop plants to manage grassy weeds. Despite its effect, the toxic residues of ATZ affect soil fertility and crop yield. Hence, the current study is focused on providing insight into the degradation mechanism of the herbicide atrazine through bacterial chemotaxis involving intermediates responsive to degradation. A bacterium was isolated from ATZ-contaminated soil and identified as Pseudomonas stutzeri based on its morphology, biochemical and molecular characterization. Upon ultra-performance liquid chromatography analysis, the free cells of isolated bacterium strain was found to utilize 174 µg/L of ATZ after 3-days of incubation on a mineral salt medium containing 200 µg/L of ATZ as a sole carbon source. It was observed that immobilized based degradation of ATZ yielded 198 µg/L and 190 µg/L by the cells entrapped with silica beads and sponge, respectively. Furthermore, the liquid chromatography-mass spectroscopy revealed that the secretion of three significant metabolites, namely, cyanuric acid, hydroxyatrazine and N- N-Isopropylammelide is responsive to the biodegradation of ATZ by the bacterium. Collectively, this research demonstrated that bacterium strains are the most potent agent for removing toxic pollutants from the environment, thereby enhancing crop yield and soil fertility with long-term environmental benefits.

Biofabricated yeast: super-soldier for detoxification of heavy metals.

The advances in nanotechnology have shown enormous impacts in environmental technology as a potent weapon for degradation of toxic organic pollutants and detoxification of heavy metals. It is either by in-situ or ex-situ adaptive strategies. Mycoremediation of environmental pollutants has been a success story of the past decade, by employing the wide arsenal of biological capabilities of fungi. Recently, the proficiency and uniqueness of yeast cell surface alterations have encouraged the generation of engineered yeast cells as dye degraders, heavy metal reduction and its recovery, and also as detoxifiers of various hazardous xenobiotic compounds. As a step forward, recent trends in research are towards developing biologically engineered living materials as potent, biocompatible and reusable hybrid nanomaterials. They include chitosan-yeast nanofibers, nanomats, nanopaper, biosilica hybrids, and TiO2-yeast nanocomposites. The nano-hybrid materials contribute significantly as supportive stabilizer, and entrappers, which enhances the biofabricated yeast cells' functionality. This field serves as an eco-friendly cutting-edge cocktail research area. In this review, we highlight recent research on biofabricated yeast cells and yeast-based biofabricated molecules, as potent heavy metals, toxic chemical detoxifiers, and their probable mechanistic properties with future application perspectives.

Warhorses in soil bioremediation: Seed biopriming with PGPF secretome to phytostimulate crop health under heavy metal stress.

The fungal symbiosis with the plant root system is importantly recognized as a plant growth promoting fungi (PGPFs), as well as elicitor of plant defence against different biotic and abiotic stress conditions. Thus PGPFs are playing as a key trouper in enhancing agricultural quality and increased crop production and paving a way towards a sustainable agriculture. Due to increased demand of food production, the over and unscientific usage of chemical fertilizers has led to the contamination of soil by organic and inorganic wastes impacting on soil quality, crops quality effecting on export business of agricultural products. The application of microbial based consortium like plant growth promoting fungi is gaining worldwide importance due to their multidimensional activity. These activities are through plant growth promotion, induction of systemic resistance, disease combating and detoxification of organic and inorganic toxic chemicals, a heavy metal tolerance ability. The master key behind these properties exhibited by PGPFs are attributed towards various secretory biomolecules (secondary metabolites or enzymes or metabolites) secreted by the fungi during interaction mechanism. The present review is focused on the multidimensional role PGPFs as elicitors of Induced systemic resistance against phytopathogens as well as heavy metal detoxifier through seed biopriming and biofortification methods. The in-sights on PGPFs and their probable mechanistic nature contributing towards plants to withstand heavy metal stress and stress alleviation by activating of various stress regulatory pathways leading to secretion of low molecular weight compounds like organic compounds, glomalin, hydrophobins, etc,. Thus projecting the importance of PGPFs and further requirement of research in developing PGPFs based molecules and combining with trending Nano technological approaches for enhanced heavy metal stress alleviations in plant and soil as well as establishing a sustainable agriculture.

A biogenic microbial biosurfactin that degrades difenoconazole fungicide with potential antimicrobial and oil displacement properties.

Surfactin is a bacterial lipopeptide and an influential biosurfactant mainly known for excellent surfactant ability. The amphiphilic nature of surfactin helps it to sustain under hydrophobic and hydrophilic conditions. In this investigation, a bacterium strain (BTKU3) that produces biosurfactant were isolated from oil-contaminated soil. Based on the blue agar plate (Bap) assay, the BTKU3 strain was found to be promising for biosurfactant production. This strain was later identified as a Lysinibacillus sp. by 16S rRNA sequencing. The characteristics of extracted bacterial surfactin were evidenced by FTIR with the presence of amine, C-H, CO, CC, esters, thiocarbonyl and asymmetric aliphatic C-H stretch molecular structural groups. Further, the extracted bacterial biosurfactant material was subjected to Liquid Chromatography-Mass Spectroscopy (LCMS), and it was identified and confirmed as surfactin with an elution time of 3.1 min and m/z value of 1034. The emulsification and oil displacement tests further proved the surfactin ability with 83% of coconut oil emulsion index and 80 % oil displacement ability with diesel, respectively. Lysinibacillus sp. BTKU3 strain also proved its efficacy in the degradation of difenoconazole by utilizing a capacity of 9.1 µg ml-1. Thus, it is inferred that the Lysinibacillus sp. BTKU3 strain plays a significant role in the production of surfactin, which positively acts as an antimicrobial agent and reduces contaminants in polluted sites.

Mitigation of organophosphorus insecticides from environment: Residual detoxification by bioweapon catalytic scavengers.

Organophosphorus insecticides (OPIs) have low persistence and are easily biodegradable in nature. The United States and India are the major countries producing OPIs of about 25% and 17% of the world, respectively. OPIs commonly used for agricultural practices occupy a major share in the global market, which leads to the increasing contamination of OPIs residues in various food chains. To overcome this issue, an enzymatic degradation method has been approved by several environmental toxic, and controlling agencies, including United States Environmental Protection Agency (USEPA). Different catalytic enzymes have been isolated and identified from various microbial sources to neutralize the toxic pesticides and/or insecticides. In this review, we have gathered information on OPIs biotransformation and their residual toxicity in the environment. Particularly, it focuses on OPIs degrading enzymes such as chlorpyrifos hydrolase, diisopropylfluorophosphatase, organophosphate acid anhydrolase, organophosphate hydrolases, and phosphotriesterases like lactonasesspecific activity either P-O link group type or P-S link group of pesticides. To summarize, the catalytic degradation of organophosphorus insecticides is not only profitable but also environmentally friendly. Hence, the enzymatic catalyst is an ultimate and super bio-weapon to mitigate or decontaminate various OPIs residues in both terrestrial and aqueous environments.

Gamma-irradiated fenugreek extracts mediates resistance to rice blast disease through modulating histochemical and biochemical changes.

The impact of gamma radiation on the activation of rice innate immunity to blast disease caused by Magnaporthe oryzae is described. In the present study, fenugreek seed extracts radiated with different doses of gamma rays viz. 5Gy, 10Gy, 15Gy, 20Gy and 25Gy were examined for their presence of biocompounds as well as for its ability to induce plant growth promotion and resistance against rice blast disease. The results of GC-MS analysis detected antimicrobial properties in methanolic extract. Enhanced germination (97%) and vigor (2718) was noticed in seeds pretreated with 20 Gy of gamma radiation in comparison with non-irradiated controls. Under greenhouse conditions, a significant disease protection of 56.7% on 3rd and 4th day after inoculation against rice blast was observed in 15Gy-irradiated rice plants challenge-inoculated with M. oryzae. Further, a significant increase in the hydrogen peroxide, phenol and lignin deposition was noticed in 20Gy-irradiated rice plants. Additionally, rice plants pretreated with 15Gy induced maximum activities of peroxidase (POX) and polyphenol oxidase (PPO) compared to untreated control plants. These findings revealed that rice plants-pretreated with gamma radiation elicit resistance against rice blast disease as well as strengthening the growth parameters by modulating cellular and biochemical defense system.

Bioactive compounds guided diversity of endophytic fungi from Baliospermum montanum and their potential extracellular enzymes.

Baliospermum montanum (Willd.) Muell. Arg, a medicinal plant distributed throughout India from Kashmir to peninsular-Indian region is extensively used to treat jaundice, asthma, and constipation. In the current study, 203 endophytic fungi representing twenty-nine species were isolated from tissues of B. montanum. The colonization and isolation rate of endophytes were higher in stem followed by seed, root, leaf and flower. The phytochemical analysis revealed 70% endophytic isolates showed alkaloids and flavonoids, 13% were positive for phenols, saponins and terpenoids. Further, these endophytes produced remarkable extracellular enzymes such as amylase, cellulase, phosphates, protease and lipase. The most promisive three endophytic fungi were identified by ITS region and secreted metabolites were identified by gas chromatography-mass spectrometry (GC-MS/MS). The GC-MS profile detected twenty-five bioactive compounds from ethyl acetate extracts. Among endophytic fungi, Trichoderma reesei isolated from flower exhibited nine bioactive compounds namely, 2-Cyclopentenone, 2-(4-chloroanilino)-4-piperidino, Oxime-methoxy-Phenyl, Methanamine N-hydroxy-N-methyl, Strychane, Cyclotetrasiloxane, Octamethyl and 1-Acetyl-20a-hydroxy-16-methylene. The endophyte, Aspergillus brasiliensis isolated from root and Fusarium oxysporum isolated from seed produced nine and seven bioactive compounds, respectively. Overall, a significant contribution of bioactive compounds was noticed from the diverse endophytic fungi associated with B. montanum and could be explored for development of novel drug with commercial values.

Exogenous priming of chitosan induces upregulation of phytohormones and resistance against cucumber powdery mildew disease is correlated with localized biosynthesis of defense enzymes.

In recent years, exploration of biopolymer-based materials to avoid hazardous chemicals in agriculture has gained enormous importance for sustainable crop improvement. In the present study, chitosan a biopolymer derived from crab-shell was used in different concentrations as priming agent to cucumber seeds and were evaluated for its effect to enhance plant growth parameters as well as its ability to induce resistance against powdery mildew disease. Among the treatments, seeds-primed with 2.5 mg/mL exhibited early seedling germination of 90% and vigour of 2665 and also remarkably enhanced the cucumber growth parameters which might be fairly attributed to the stimulation of phytohormones content in primed plants over the controls. More importantly, under greenhouse conditions a significant induced disease protection of 66.6% against powdery mildew disease was noticed in chitosan-pretreated plants at 2.5 mg/mL. The induced resistant plants also showed a significant deposition of lignin, callose and H2O2. Notably, polyphenol oxidase, phenylalanine ammonia-lyase, peroxidase and glucanase defense-responsive enzymes were upregulated in chitosan-primed plants. Considered together, these results determine that the susceptible cucumber cultivar elicits immunity after perception of priming with chitosan to upregulate phytohormones and synthesize defense-responsive enzymes, thereby induce resistance against powdery mildew disease and strengthen the growth-promotion of cucumber plants.

Isolation and characterization of nutrient dependent pyocyanin from Pseudomonas aeruginosa and its dye and agrochemical properties.

Pyocyanin is a blue green phenazine pigment produced in large quantities by active cultures of Pseudomonas aeruginosa, with advantageous applications in medicine, agriculture and for the environment. Hence, in the present study, a potent bacterium was isolated from agricultural soil and was identified morphologically and by 16S rRNA sequencing as P. aeruginosa (isolate KU_BIO2). When the influence of nutrient supplements in both King's A and Nutrient media as amended was investigated, an enhanced pyocyanin production of 2.56 µg ml-1 was achieved in King's A medium amended with soya bean followed by 1.702 µg ml-1 of pyocyanin from the nutrient medium amended with sweet potato. Purified pyocyanin was characterized by UV-Vis Spectrophotometer and Fourier-Transform Infrared spectroscopy (FTIR). Furthermore, Liquid Chromatography Mass Spectrum (LCMS) and Nuclear Magnetic Resonance (NMR) confirmed its mass value at 211 and as N-CH3 protons resonating at 3.363 ppm as a singlet respectively. The isolated pyocyanin displayed remarkable dye property by inducing color change in cotton cloth from white to pink. Lastly, the antifungal activity of test pyocyanin showed inhibition of growth of rice blast fungus, Magnaporthe grisea and bacterial blight of rice, Xanthomonas oryzae at concentrations of 150 and 200 ppm, respectively. Thus, this investigation provides evidence for diverse actions of pyocyanin which are nutrient dependent and are capable of acting on a large scale, by utilizing microbes existing in agriculture wastes, and thus could be used as an alternative source in the making of natural textile dyes with strong durability and a broad spectrum of ecofriendly agrochemicals.

Detection and determination of stability of the antibiotic residues in cow's milk.

In the present study, antibiotic residues were detected in milk samples collected from the dairy herds located in Karnataka, India, by microbiological assay. Subsequently, the detected antibiotics were identified as azithromycin and tetracycline, by high-performance liquid chromatography, further both the antibiotics detected in the cow milk samples were found to be at high concentration (9708.7 and 5460 µg kg-1, respectively). We then investigated the effects of temperature and pH on the stabilities of azithromycin and tetracycline to determine the degradation rate constant k using first-order kinetic equation. Results indicated that significant reduction in stability and antibacterial activity of azithromycin solution when subjected to 70 and 100°C for 24 h. While stability of tetracycline was significantly reduced when subjected to 70 and 100°C for 24 h. However no significant reduction in antibacterial activity of tetracycline was observed at respective temperatures when compared with that of control. In addition, the stabilities of azithromycin and tetracycline were found to be decreased in acidic pH 4-5. The results of the present study revealed the high risk of contamination of milk sample with veterinary antibiotics and also demonstrated the effect of temperature and pH on stability of antibiotics. Therefore the study suggest that the qualitative and quantitative screening of milk for the presence of antibiotics need to be strictly performed to ensure safe drinking milk for consumers.

Physiological adaptation and spectral annotation of Arsenic and Cadmium heavy metal-resistant and susceptible strain Pseudomonas taiwanensis.

In the present study, the 16S-rRNA sequencing of heavy metal-resistant and susceptible bacterial strains isolated from the industrial and agriculture soil showed resemblance with Pseudomonas taiwanensis. Based on the growth rate, two bacterial strains SJPS_KUD54 and KUD-MBBT4 exhibited 10 ppm tolerance to Arsenic and Cadmium. These two heavy metals caused, a significant increase in stress enzymes like superoxide dismutase, catalase and glutathione S-transferase activities in SJPS_KUD54 when compared to KUD-MBBT4. Following heavy metal treatment, the atomic-force-microscopy observations showed no change in the cell-wall of SJPS_KUD54, whereas the cell-wall of KUD-MBBT4 got ruptured. Moreover, the protein-profile of SJPS_KUD54 treated with heavy metals exhibited varied patterns in comparison with untreated control. In addition, the accumulation of hydroxyl, thiol and amides were found in the SJPS_KUD54 relative to its control. Furthermore, the resistant SJPS_KUD54 strain showed a remarkable bioaccumulation properties to both Arsenic and Cadmium. Thus, it is inferred that the growth rate, stress enzymes and functional-groups play a significant role in the physiological-adaption of SJPS_KUD54 during stress conditions, which is positively involved in the prevention or repair mechanism for reducing the risks caused by heavy metal stress.

Influence of triazole pesticides on tillage soil microbial populations and metabolic changes.

Pesticides are generally applied to agricultural soil to control crop diseases, with the critical goal of preventing yield loss and optimizing economic returns. However, the excessive utilization of pesticides in agriculture is a severe environmental and human health problem. In the present study, effect of triazole based fungicide, propiconazole on soil physicochemical properties, stimulatory impact of propiconazole residue on soil microbial communities and soil enzyme activities (phosphatise and urease) in contaminated red sandy loam and deep black soils of paddy (Oryza sativa L.) fields at different concentrations were evaluated. The liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis showed high levels of propiconazole residues of 434 and 426 ng g-1 in red sandy loam and deep black soils, respectively. A total of 27 bacterial and 18 fungal strains were isolated from propiconazole-contaminated soils, identified by molecular markers and had their sequences deposited in the GenBank. We observed a pronounced increase in soil microbial growth, and urease and phosphatase activities at lower propiconazole concentrations up to 2 weeks post-treatment. On the contrary, the microbial growth and urease and phosphatase activities were relentlessly reduced at higher propiconazole concentrations and a long incubation period after 2 to 4 weeks of post-treatment. Overall, it was concluded that the concentration-dependency and incubation period of propiconazole would enhance the growth and activity of soil microbes and enzymes respectively and the higher application will be detrimental to the soil health.

Computational Modeling of the Staphylococcal Enterotoxins and Their Interaction with Natural Antitoxin Compounds

Staphylococcus aureus is an opportunistic bacterium that produces various types of toxins, resulting in serious food poisoning. Staphylococcal enterotoxins (SEs) are heat-stable and resistant to hydrolysis by digestive enzymes, representing a potential hazard for consumers worldwide. In the present study, we used amino-acid sequences encoding SEA and SEB-like to identify their respective template structure and build the three-dimensional (3-D) models using homology modeling method. Two natural compounds, Betulin and 28-Norolean-12-en-3-one, were selected for docking study on the basis of the criteria that they satisfied the Lipinski's Rule-of-Five. A total of 14 and 13 amino-acid residues were present in the best binding site predicted in the SEA and SEB-like, respectively, using the Computer Atlas of Surface Topology of Proteins (CASTp). Among these residues, the docking study with natural compounds Betulin and 28-Norolean-12-en-3-one revealed that GLN43 and GLY227 in the binding site of the SEA, each formed a hydrogen-bond interaction with 28-Norolean-12-en-3-one; while GLY227 residue established a hydrogen bond with Betulin. In the case of SEB-like, the docking study demonstrated that ASN87 and TYR88 residues in its binding site formed hydrogen bonds with Betulin; whereas HIS59 in the binding site formed a hydrogen-bond interaction with 28-Norolean-12-en-3-one. Our results demonstrate that the toxic effects of these two SEs can be effectively treated with antitoxins like Betulin and 28-Norolean-12-en-3-one, which could provide an effective drug therapy for this pathogen.

Biodegradation of propiconazole by newly isolated Burkholderia sp. strain BBK_9.

The isolation of propiconazole (PCZ) degrading bacterium BBK_9 strain was done from paddy soil, and it was identified as Burkholderia sp. based on the morphological characteristics and biochemical properties combined with 16S rRNA gene sequencing analysis. It has been seen that the factors such as temperature and pH influence the biodegradation process. The role of plasmid was studied in the degradation process by plasmid curing method. The PCZ acts as the sole carbon source and as energy substrate which can be utilized by the strain for its growth in Mineral salt medium and degraded 8.89 µg ml-1 of PCZ at 30 °C and pH 7 within 4 days. During the bioconversion process of PCZ, three metabolite were formed such as 1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl) ethanone, 1-[2-(4-chlorophenyl) ethyl]-1H-1,2,4-triazole and 1-ethyl-1H-1,2,4-triazole. The LD50 value of BBK_9 strain was determined with acridine orange which resulted in 40 µg ml-1 at cell density of 0.243 at 660 nm. Furthermore, plasmid curing was done using LD50 concentration and from that three plasmids got cured in the sixth generation. It was found that, cured strain was able to degrade 7.37 µg ml-1 of PCZ, indicating the plasmid encoded gene were not responsible for the PCZ degradation. On the source of these outcomes, strain BBK_9 can be used as potential strain for bioremediation of contaminated sites.