Co-existence of halo-tolerant Pseudomonas fluorescens and Enterococcus hirae with multifunctional growth promoting traits to ameliorate salinity stress in Vigna radiata.

Soil salinization has become a prominent obstacle in diverse arid and semi-arid region damaging agricultural productivity globally. From this perspective, present investigation was aimed to compare the potential compatible consortium of bio-inoculants for improving Plant Growth Promoting (PGP) attributes, anti-oxidative enzymes, grain yield and profitability of Vigna radiata in saline soil conditions. A total of 101 rhizobacterium isolated from salt affected regions of Punjab, India were screened for their ability to induce salt tolerance, multifunctional PGP traits and antagonistic activities. The 16S rRNA sequencing identified the strains LSMR-29 and LSMRS-7 as Pseudomonas flourescens and Enterococcus hirae, respectively. In-vitro compatible halo-tolerant dual inoculant (LSMR-29 + LSMRS-7) as bio-inoculants mitigated salt stress in Vigna radiata (spring mungbean) seedling with improved seed germination, biomass and salt tolerance index together with the presence of nifH, acds, pqq and ipdc gene under salinity stress as compared to single inoculants. Further, the potential of single and dual bio-inoculants were also exploited for PGP attributes in pot and field experiments. Results indicated that a significant improvement in chlorophyll content (2.03 fold), nodulation (1.24 fold), nodule biomass (1.23 fold) and leghemoglobin content (1.13 fold) with dual inoculant of LSMR-29 + LSMRS-7 over the LSMR-29 alone. The concentrations of macro & micronutrients, proline, soil enzyme activities i.e. soil dehydrogenase, acid & alkaline phosphatases and antioxidant enzymes such as superoxide dismutase, catalase and peroxidase also found to be high for LSMR-29 + LSMRS-7 as compared to un-inoculated control. The high grain yield thereby leading to Benefit: Cost (B: C) ratio at field scale was indicative of the commercial use bio-inoculants under salt affected Vigna radiata (spring mungbean) to improvement of productivity and soil health. The current finding reveals a co-inoculation of halo-tolerating Pseudomonas fluorescens and Enterococcus hirae containing ACC deaminase could prove to be novel approach for inducing salt tolerance and improving productivity of Vigna radiata (spring mungbean).

Elicitation of E-waste (acrylonitrile-butadiene styrene) enriched soil bioremediation and detoxification using Priestia aryabhattai MGP1.

Given the rise in both usage and disposal of dangerous electronics, there is a catastrophic rise in assemblage of electronic waste (e-waste). E-waste including various plastic resins are among the most frequently discarded materials in electronic gadgets. In current digital era, managing e-waste has become universal concern. From the viewpoint of persisting lacuna of e-waste managing methods, the current study is designed to fabricate an eco-friendly e-waste treatment with native soil bacteria employing an enrichment culture method. In the presence of e-waste, indigenous soil microbes were stimulated to degrade e-waste. Microbial cultures were isolated using enrichment medium containing acrylonitrile-butadiene styrene (ABS) as the primary carbon source. Priestia aryabhattai MGP1 was found to be the most dominant e-polymer degrading bacterial isolate, as it was reported to degrade ABS plastic in disposed-off television casings. Furthermore, to increase degradation potential of MGP1, Response Surface Methodology (RSM) was adopted which resulted in optimized conditions (pH 7, shaking-speed 120 rpm, and temperature 30 °C), for maximum degradation (18.88%) after 2 months. The structural changes induced by microbial treatment were demonstrated by comparing the findings of Field emission scanning electron microscopy (FESEM) images and Fourier Transform Infrared (FTIR) spectra confirming the disappearance of ≡ C─H peaks along with C-H, C=C and C ≡N bond destabilization following degradation. Energy-dispersive X-ray (EDX) analyzers of the native and decomposed e-polymer samples revealed a considerable loss in elemental weight % of oxygen by 8.4% and silica by 0.5%. Magnesium, aluminium and chlorine which were previously present in the untreated sample, were also removed after treatment by the bacterial action. When seeds of Vigna radiata were screened using treated soil in the presence of both e-waste and the chosen potent bacterial strain, it was also discovered that there was reduced toxicity in terms of improved germination and growth metrics as a phytotoxicity criterion.

Eco-friendly detoxification of hazardous Congo red dye using novel fungal strain Trametes flavida WTFP2: Deduced enzymatic biomineralization process through combinatorial in-silico and in-vitro studies.

Growing textile industry is a major global concern, owing to the presence of recalcitrant hazardous pollutants, like synthetic dyes in discharged effluents. To explore new bioresources for mycoremediation, a high laccase-producing novel white-rot fungus (WRF), Trametes flavida WTFP2, was employed. T. flavida is an underexplored member of Polyporales. Using bioinformatic tools, 8 different cis-acting RNA elements were identified in the 5.8 S ITS gene sequence, where CRISPR (CRISPR-DR15), sRNA (RUF1), and snoRNA (ceN111) are uniquely present. Molecular docking was adopted to predict the catalytic interaction of chosen toxic diazo colorant, Congo red (CR), with four dye-degrading enzymes (laccase, lignin peroxidase, azoreductase, and aryl alcohol oxidase). With 376.41 × 103 U/L laccase production, novel WRF exhibited dye-decolorization potential. WTFP2 effectively removed 99.48 ± 0.04% CR (100 mg/L) and demonstrated remarkable recyclability and persistence in consecutive remediation trials. Mycelial dye adsorption was not only substantial driver of colorant elimination; decolorization using active T. flavida was regulated by enzymatic catalysis, as outlined by in-vitro growth, induction of extracellular enzymes, and FESEM. Fifteen metabolites were identified using HRLCMS-QTOF, and novel CR degradation pathway was proposed. Furthermore, microbial and phyto-toxicity tests of metabolites suggested complete detoxification of toxic dye, making the process clean, green, and economically sustainable.

SAR based Review on Diverse Heterocyclic Compounds with Various Potential Molecular Targets in the Fight against COVID-19: A Medicinal Chemist Perspective.

Coronavirus disease (COVID-19) was reported to be transmitted from bats to humans and, became a pandemic in 2020. COVID-19 is responsible for millions of deaths worldwide and still, the numbers are increasing. Further, despite the availability of vaccines, mutation in the virus continuously poses a threat of re-emergence of the more lethal form of the virus. So far, the repurposing of drugs has been exercised heavily for the identification of therapeutic agents against COVID-19, which led FDA to approve many drugs for the same e.g., remdesivir, favipiravir, ribavirin, etc. The anti-COVID drugs explored via other approaches include nirmatrelvir (used in combination with ritonavir as Paxlovid), tixagevimab and cilgavimab (both used in combination with each other) and others. However, these approved drugs failed to achieve a significant clinical outcome. Globally, natural bioactive have also been explored for anti-COVID-19 effects, based on their traditional medicinal values. Although the clinical findings suggest that FDA-approved drugs and natural bioactives can help reducing the overall mortality rate but the significant clinical outcome was not achieved. Therefore, the focus has been shifted towards new drug development. In line with that, a lot of work has been done and still going on to explore heterocyclic compounds as potent anti- COVID-19 drugs. Several heterocyclic scaffolds have been previously reported with potent antiinflammatory, anticancer, anti-viral, antimicrobial and anti-tubercular effects. Few of them are under consideration for clinical trials whereas others are under preclinical investigation. Hence, this review discusses the evidence of rationally designed and tested heterocyclic compounds acting on different targets against COVID-19. The present article will help the researches and will serve as a pivotal resource in the design and development of novel anti-COVID-19 drugs.

Nano-biofertilizers as bio-emerging strategies for sustainable agriculture development: Potentiality and their limitations.

Nanotechnology is a burgeoning revolutionary technology in the 21st century. Climate emergencies caused by natural or anthropogenic activities have tragically consequential repercussions on agricultural output worldwide. Modern cropping systems profoundly rely on synthetic fertilizers to deliver necessary nutrients, yet their prolonged and persistent administration is hazardous to the environment, soil fertility, and nutritional dynamics of the rhizospheric microbiome. By addressing the drawback of physico-chemically synthesized nano-dimensioned fertilizer, this review emphasizes on integrating nanoparticles and biofertilizers conjointly as nano-biofertilizers (NBF) which can safeguard global food security, in light of the population surge. Inoculation with nanoparticles and biofertilizers strengthens plant growth and stress tolerance. However, combined together (NBF), they have emerged as a more economically and environmentally sustainable, highly versatile, and long-lasting agriculture tool. Microbe-based green synthesis using the encapsulation of inorganic nanoparticles of Si, Zn, Cu, Fe, Ni, Ti, and Ag as well as organic materials, including chitosan, cellulose, and starch, to formulate NBFs can eliminate the constraints of conventional fertilizer contamination. The application of NBFs is in its infancy in agriculture, yet it has promising potential for transforming traditional farming techniques into smart agriculture, compared to any of the existing strategies. From this perspective, this review is an attempt to provide a comprehensive understanding of the formulations, fabrication, and characterization of NBFs while unraveling the underlying mechanisms of plant-NBF interactions along with their contribution to climate change-induced biotic and abiotic stress tolerance. We substantially summarize the latest advancements of field applications of NBFs for precision farming. Moreover, we critically revised their applications in agro-ecosystems according to the current literature, while also discussing the bottlenecks and future trends for developing potent NBFs.

Role of NLRP3 Inflammasome and Its Inhibitors as Emerging Therapeutic Drug Candidate for Alzheimer's Disease: a Review of Mechanism of Activation, Regulation, and Inhibition.

Alzheimer's disease (AD) is one of the most prevalent neurodegenerative disorders. The etiology and pathology of AD are complicated, variable, and yet to be completely discovered. However, the involvement of inflammasomes, particularly the NLRP3 inflammasome, has been emphasized recently. NLRP3 is a critical pattern recognition receptor involved in the expression of immune responses and has been found to play a significant role in the development of various immunological and neurological disorders such as multiple sclerosis, ulcerative colitis, gout, diabetes, and AD. It is a multimeric protein which releases various cytokines and causes caspase-1 activation through the process known as pyroptosis. Increased levels of cytokines (IL-1ß and IL-18), caspase-1 activation, and neuropathogenic stimulus lead to the formation of proinflammatory microglial M1. Progressive researches have also shown that besides loss of neurons, the pathophysiology of AD primarily includes amyloid beta (Aß) accumulation, generation of oxidative stress, and microglial damage leading to activation of NLRP3 inflammasome that eventually leads to neuroinflammation and dementia. It has been suggested in the literature that suppressing the activity of the NLRP3 inflammasome has substantial potential to prevent, manage, and treat Alzheimer's disease. The present review discusses the functional composition, various models, signaling molecules, pathways, and evidence of NLRP3 activation in AD. The manuscript also discusses the synthetic drugs, their clinical status, and projected natural products as a potential therapeutic approach to manage and treat NLRP3 mediated AD.

Plant growth-promoting rhizobacteria: Salt stress alleviators to improve crop productivity for sustainable agriculture development.

Soil salinity, a growing issue worldwide, is a detrimental consequence of the ever-changing climate, which has highlighted and worsened the conditions associated with damaged soil quality, reduced agricultural production, and decreasing land areas, thus resulting in an unsteady national economy. In this review, halo-tolerant plant growth-promoting rhizo-microbiomes (PGPRs) are evaluated in the salinity-affected agriculture as they serve as excellent agents in controlling various biotic-abiotic stresses and help in the augmentation of crop productivity. Integrated efforts of these effective microbes lighten the load of agro-chemicals on the environment while managing nutrient availability. PGPR-assisted modern agriculture practices have emerged as a green strategy to benefit sustainable farming without compromising the crop yield under salinity as well as salinity-affected supplementary stresses including increased temperature, drought, salinity, and potential invasive plant pathogenicity. PGPRs as bio-inoculants impart induced systemic tolerance (IST) to plants by the production of volatile organic compounds (VOCs), antioxidants, osmolytes, extracellular polymeric substances (EPS), phytohormones, and ACC-deaminase and recuperation of nutritional status and ionic homeostasis. Regulation of PGPR-induced signaling pathways such as MAPK and CDPK assists in salinity stress alleviation. The "Next Gen Agriculture" consists of the application of designer crop microbiomes through gene editing tools, for instance, CRISPR, and engineering of the metabolic pathways of the microbes so as to gain maximum plant resistance. The utilization of omics technologies over the traditional approaches can fulfill the criteria required to increase crop yields in a sustainable manner for feeding the burgeoning population and augment plant adaptability under climate change conditions, ultimately leading to improved vitality. Furthermore, constraints such as the crop specificity issue of PGPR, lack of acceptance by farmers, and legal regulatory aspects have been acknowledged while also discussing the future trends for product commercialization with the view of the changing climate.

Differential proteomic analysis under pesticides stress and normal conditions in Bacillus cereus 2D.

A potential pesticide degrading bacterial isolate (2D), showing maximum tolerance (450 mg∙L-1) for cypermethrin, fipronil, imidacloprid and sulfosulfuron was recovered from a pesticide contaminated agricultural field. The isolate degraded cypermethrin, imidacloprid, fipronil and sulfosulfuron in minimal salt medium with 94, 91, 89 and 86% respectively as revealed by high performance liquid chromatography (HPLC) and gas chromatography (GC) analysis after 15 days of incubation. Presence of cyclobutane, pyrrolidine, chloroacetic acid, formic acid and decyl ester as major intermediate metabolites of cypermethrin biodegradation was observed in gas chromatography mass spectrometry (GC-MS) analysis. Results based on 16S rDNA sequencing, and phylogenetic analysis showed maximum similarity of 2D with Bacillus cereus (Accession ID: MH341691). Stress responsive and catabolic/pesticide degrading proteins were over expressed in the presence of cypermethrin in bacteria. Enzymatic kinetics of laccase was deduced in the test isolate under normal and pesticide stress conditions which suggested that the production of enzyme was induced significantly in pesticide stress (163 µg.µL-1) as compare to normal conditions(29 µg.µL-1) while the Km value was decreased in pesticides stress condition (Km = 10.57 mM) and increases in normal condition (Km = 14.33 mM).Amplification of laccase gene showed a major band of 1200bp. The present study highlights on the potential of 2D bacterial strain i.e., high tolerance level of pesticide, effective biodegradation rate, and presence of laccase gene in bacterial strain 2D, could become a potential biological agent for large-scale treatment of mixture of pesticide (cypermethrin, fipronil, imidacloprid and sulfosulfuron) in natural environment (soil and water).

Occurrence of multidrug resistant Escherichia coli in groundwater of Brij region (Uttar Pradesh) and its public health implications.

AIM: The study evaluates the microbial as well as physicochemical pollution of groundwater of Brij region of Uttar Pradesh, a major tourist destination in the country along with estimating the drug resistance evident in the isolated Escherichia coli. MATERIALS AND METHODS: A total of 60 samples of groundwater were collected from six different sites and assessed for physicochemical (pH, color, taste, turbidity, total dissolved solids [TDS], total hardness [TH], chlorides, fluorides, nitrates, and iron) and microbiological parameters (standard plate count [SPC], most probable number test [MPN], E. coli). RESULTS: A majority of the samples were found to be out of the range for most of the parameters except iron. Particularly, high values of TDS (up to 9000 ppm), TH (1500 mg/L), chlorides (3250 mg/L), fluorides (2.5 mg/L), and nitrates (100.2 mg/L) were observed at most of the sites in the region highlighting the fact that groundwater of the area is not potable. Samples were turbid and salty to taste. High SPC values, up to 3500 colony-forming unit/ml and coliforms beyond BIS range were found in 40% samples suggesting gross microbial contamination. Only 2 sites (G3 and G5) had low MPN values. Overall 16 (26.67%) E. coli were isolated with 3 (18.75%) producing red colonies on conge red agar, hence supposed to be pathogenic. No E. coli O157:H7 was isolated. High antimicrobial resistance was observed against amoxicillin and erythromycin, whereas E. coli isolates were sensitive toward cefotaxime-clavulanic acid and imipenem. 12 isolates (75%) were multidrug resistant (MDR) with MDR index >20%, and 2 isolates (12.5%) were found to be extended spectrum beta-lactamases positive. CONCLUSION: Groundwater is considered to be a safe option for potable water but it is obvious from the findings of this study that considerable physicochemical and microbial contamination is there in groundwater samples of Brij region. The occurrence of MDR E. coli in these waters is a matter of great public health concern.

A study on association of virulence determinants of verotoxic Escherichia coli isolated from cattle calves.

AIM: The present study was conducted to find the association among virulence determinants of verotoxic Escherichia coli (VTEC) isolated from cattle calf feces. MATERIALS AND METHODS: A total of 216 cattle calf fecal samples were collected aseptically and processed under required conditions for the isolation of E. coli. The isolates were further subjected to multiplex polymerase chain reaction (mPCR) for the detection of virulent genes. All the VTEC isolates were serotyped at the Central Research Institute, Kasauli, Himachal Pradesh. The VTEC isolates were observed for the enterohemolysin production on washed sheep blood agar (wSBA). RESULTS: A total of 177 presumptive E. coli were isolated from 216 calf fecal samples revealing an overall prevalence of E. coli to be 81.94%. A total of 32 (14.81%) isolates were detected as VTEC through mPCR. The prevalence of verotoxin genes vt1, vt2, and combination of vt1+vt2 in the VTEC isolates was found to be 12 (37.5%), 14 (43.75%), and 6 (18.75%), respectively. Other virulent genes eaeA and hlyA were found in 6 and 11 VTEC strains with prevalence values of 18.75% and 34.37%, respectively. A total of 13 different O serogroups were revealed in serotyping of 32 VTEC isolates. Out of 32 VTEC strains, only 26 (81.25%) were enterohemolytic on wSBA as they produced the characteristic small, turbid zone of hemolysis around the streaking line. Although enterohemolysin production has been attributed to the presence of hlyA gene, only 11 of 26 enterohemolysin producing VTEC were found to be harboring the hlyA gene (11/26) 42.03%. CONCLUSION: The present study concludes that there might be an association between the presence of verotoxin genes and enterohemolysin production in VTEC group of E. coli.

Development of a sandwich ELISA for the detection of bovine herpesvirus type 1.

OBJECTIVE: To develop a standard enzyme-linked immunosorbent assay (ELISA) for the detection of bovine herpesvirus type 1 (BHV-1). METHODS: The assay was based on hyperimmune rabbit and guinea pig antisera raised against purified BHV-1. Polyethylene glycol precipitation and sucrose density gradient methods were adopted for viral concentration and purification. Antisera were raised using Freund's adjuvant followed by extraction of IgG of high purity. RESULTS: Optimum antisera dilutions as determined by titrations were chosen as 1:4 000, whereas the conjugate was used at 1:2 000 dilution. Using 95 clinical specimens, the ELISA test showed a sensitivity and specificity of 91.90 % and 93.10 %, respectively when compared to PCR. The cut-off value was fixed at 0.15 (A(490)) and a P/N ratio of >1.30 indicated a significant positive reaction. CONCLUSIONS: The results have demonstrated that this ELISA could efficiently detect BHV-1 and can be used as an important diagnostic tool.

Immunochemical characterization of antigens of Brucella canis and their use in seroprevalence study of canine brucellosis

OBJECTIVE@#To explore immunochemical characterization of antigens of Brucella canis (B. canis), and the use in seroprevalence study of canine brucellosis.@*METHODS@#External hot phosphate buffer saline extract (HPBSE) and internal sonicated (SA) antigens were prepared from B. canis strain MEX 51 and immunochemically characterized. These antigens were used to test 527 serum samples of dogs by 2-mercaptoethanol-tube agglutination test (2 ME-TAT), agar gel immunodiffusion test (AGID), dot-ELISA and indirect enzyme-linked immunosorbent assay (I-ELISA) to assess the seroprevalence of canine brucellosis.@*RESULTS@#The protein content of HPBSE and SA antigens was 0.387 mg/mL and 0.195 mg/mL, respectively, whereas carbohydrate content was 0.174 mg/mL and 0.150 mg/mL, respectively. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (12.5%) of HPBSE and SA, revealed 6 and 8 visible peptide bands ranging from 18-80 kDa and 12-45 kDa, respectively. Western blot analysis showed immunodominant bands of MW 12, 28, 39 and 45 kDa for HPBSE and 20-24 kDa for SA. The AGID revealed HPBSE as more specific antigen than SA but both I-ELISA and dot-ELISA indicated SA antigen to be more specific and reliable than HPBSE. The seroprevalence of canine brucellosis was 2.27% by 2ME-TAT, 1.5% by AGID, 3.03% by dot-ELISA and 16.12% by I-ELISA.@*CONCLUSIONS@#On the basis of the results of present study, we concluded that HPBSE is suitable antigen for AGID, which is more specific; whereas SA antigen is suitable for I-ELISA, which is highly sensitive. Therefore, initial screening of serum samples should be carried out by I-ELISA followed by confirmation with AGID.