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Abstract For the application of biochar in restoring pesticide-contaminated soils and minimizing the risk associated with their uptake in plants, it is crucial to understand the biochar impact on soil biological activities and dissipation and accumulation of pesticides in plant and soil systems. In this study, the effect of Mentha-distilled waste-derived biochar was investigated on chlorpyrifos and atrazine contaminated sandy loam soil. The four application rates of atrazine (2, 4, 6, and 8 mg kg−1) and chlorpyrifos (2, 4, 6, and 12 mg kg−1) and a single application rate of biochar (4%) were used in this study. The degradation of pesticides, the diversity of the bacterial community, and enzymatic activities (alkaline phosphatase, dehydrogenase, arylsulfatase, phenol oxidase, urease activity and N-acetyl glucosaminidase) were examined in soil. The uptake of two pesticides and their effect on growth and stress parameters were also investigated in plants (A. paniculata). The dissipation of chlorpyrifos and atrazine followed simple first-order kinetics with a half-life of 6.6–74.6 and 21–145 days, respectively. The presence of deisopropyl atrazine desethyl atrazine (metabolites of atrazine) and 3,5,6-trichloro-2-pyridinol (a metabolite of chlorpyrifos) was observed in soil and plant tissues. Biochar application significantly (p = 0.001) enhanced the degradation rate of chlorpyrifos and atrazine leading to the lower half-life of chlorpyrifos and atrazine in soil. A significant reduction (p = 0.001) in the uptake of chlorpyrifos and atrazine and alteration in their binding affinity and uptake rate in plant tissues was observed in biochar treatments. The incorporation of biochar improved chlorpyrifos/atrazine degrader and plant growth-promoting bacterial genera such as Balneimonas, Kaistobacter, Rubrobacter, Ammoniphilus, and Bacillus. The upregulation of functional genes associated with nucleotide, energy, carbohydrate, amino acid metabolism, xenobiotic biodegradation, and metabolism: atrazine degradation was observed in biochar treatments. The biochar amendments significantly (p = 0.001) reduced the plant’s uptake velocity (Vmax) and affinity (Km) of chlorpyrifos and atrazine. These results delineated that Mentha-distilled waste-derived biochar can potentially remediate chlorpyrifos and atrazine contaminated soils and ensure the safety of plants for consumption. Graphical Abstract

Background: Childhood epilepsy is a prevalent neurological disorder. Imaging, especially MRI of the brain, plays a pivotal role in diagnosing the underlying cause. This study aimed to assess the frequency of causative factors of epilepsy detected in MRI. Materials and Methods: This hospital-based prospective observational study was conducted in the Radiology Department at Rajendra Institute of Medical Sciences (RIMS), Ranchi, Jharkhand, India from November 2021 to October 2022 in 100 children of 0 to 12 years of age referred from Pediatrics department for an MRI brain scan. MRI of the brain was performed in all cases and findings were analyzed and causes of epilepsy were assessed. Magnetic resonance spectroscopy (MRS) was also done when required for confirmation of diagnosis. Results: Positive findings in MRI were detected in 87% of children, and no abnormalities were detected in 13%. The majority of children belonged to the age group of 10-12 years (37%) and were predominantly males (66%). The most common cause of epilepsy was infections (27%) followed by hypoxic ischemic encephalopathy (22%). Tuberculoma was the most common infective cause of epilepsy in 59.3%. These were further followed by temporal lobe epilepsy and congenital malformations (11% each). The rest were other miscellaneous and idiopathic causes. Conclusion: MRI findings were specific to various conditions, helping in the localization and characterization of etiologies and playing a significant role in the evaluation of children who were newly diagnosed with epilepsy, especially those with partial seizures. Recommendation: Further research with a larger sample size and meta-analysis is recommended for more conclusive results.

Medicago sativa L. (alfalfa), a leguminous crop, is vital globally for forage, crop rotation, and soil fertility. Though, the microbiome of alfalfa grown in uncultivated land has not been studied yet. Here, we employed the high-throughput 16S rRNA and nifH gene amplicon sequencing to unravel the rhizosphere microbiome and nodules endosymbionts of alfalfa grown in poor lawn and coastal sandy soils. The root nodule of alfalfa of coastal sands and poor lawn soil, Sinorhizobia/Ensifer accounted for 80.599% and 67.727% of the total microbiome, respectively. The class Actinobacteria (except Frankiales) and order Rhizobiales, Sphingomonadales, Burkholderiales, Pseudomonas, and Cytophagia were found significantly enriched. Culturable studies displayed the dominance of Bacillus followed by Arthrobacter, Microbacterium, and Streptomyces. Hydroponics studies revealed the efficiency of cultured isolates for nodulation and plant growth attributes. Overall studies decipher the soil indigenous microbiome associated with root nodule and rhizosphere of Alfalfa grown in uncultivated coastal and sandy land.

Chickpea (Cicer arietinum L.), encompassing the desi and kabuli varieties, is a beloved pulse crop globally. Its cultivation spans over fifty countries, from the Indian subcontinent and southern Europe to the Middle East, North Africa, the Americas, Australia, and China. With a rich composition of carbohydrates and protein, constituting 80% of its dry seed mass, chickpea is also touted for its numerous health benefits, earning it the title of a ‘functional food’. In the past two decades, research has extensively explored the rhizobial diversity associated with chickpea and its breeding in various countries across Europe, Asia, and Oceania, aiming to understand its impact on the sustainable yield and quality of chickpea crops. To date, four notable species of Mesorhizobium—M. ciceri, M. mediterraneum, M. muleiense, and M. wenxiniae—have been reported, originally isolated from chickpea root nodules. Other species, such as M. amorphae, M. loti, M. tianshanense, M. oportunistum, M. abyssinicae, and M. shonense, have been identified as potential symbionts of chickpea, possibly acquiring symbiotic genes through lateral gene transfer. While M. ciceri and M. mediterraneum are widely distributed and studied across chickpea-growing regions, they remain absent in China, where M. muleiense and M. wenxiniae are the sole rhizobial species associated with chickpea. The geographic distribution of chickpea rhizobia is believed to be influenced by factors such as genetic characteristics, competitiveness, evolutionary adaptation to local soil conditions, and compatibility with native soil microbes. Inoculating chickpea with suitable rhizobial strains is crucial when introducing the crop to new regions lacking indigenous chickpea rhizobia. The introduction of a novel chickpea variety, coupled with the effective use of rhizobia for inoculation, offers the potential not only to boost the yield and seed quality of chickpeas, but also to enhance crop productivity within rotation and intercropped systems involving chickpea and other crops. Consequently, this advancement holds the promise to drive forward the cause of sustainable agriculture on a global scale.

Abstract Background Myxococcus xanthus DK1622 is a model system for studying multicellular development, predation, cellular differentiation, and evolution. Furthermore, it is a rich source of novel secondary metabolites and is widely used as heterologous expression host of exogenous biosynthetic gene clusters. For decades, genetic modification of M. xanthus DK1622 has mainly relied on kanamycin and tetracycline selection systems. Results Here, we introduce an alternative selection system based on chloramphenicol (Cm) to broaden the spectrum of available molecular tools. A chloramphenicol-resistant growth phase and a chloramphenicol-susceptible growth phase before and after chloramphenicol-induction were prepared, and later sequenced to identify specific genes related to chloramphenicol-repercussion and drug-resistance. A total of 481 differentially expressed genes were revealed in chloramphenicol-resistant Cm5_36h and 1920 differentially expressed genes in chloramphenicol-dormant Cm_8h. Moreover, the gene expression profile in the chloramphenicol-dormant strain Cm_8h was quite different from that of Cm5_36 which had completely adapted to Cm, and 1513 differentially expression genes were identified between these two phenotypes. Besides upregulated acetyltransferases, several transporter encoding genes, including ABC transporters, major facilitator superfamily transporters (MFS), resistance-nodulation-cell division (RND) super family transporters and multidrug and toxic compound extrusion family transporters (MATE) were found to be involved in Cm resistance. After the knockout of the most highly upregulated MXAN_2566 MFS family gene, mutant strain DK-2566 was proved to be sensitive to Cm by measuring the growth curve in the Cm-added condition. A plasmid with a Cm resistance marker was constructed and integrated into chromosomes via homologous recombination and Cm screening. The integration efficiency was about 20% at different concentrations of Cm. Conclusions This study provides a new antibiotic-based selection system, and will help to understand antibiotic resistance mechanisms in M. xanthus DK1622.

Soil salinization limits crop growth and yield in agro-ecosystems worldwide by reducing soil health and altering the structure of microbial communities. Salt-tolerant plant growth-promoting rhizobacteria (PGPR) alleviate plant salinity stress. Wild soybean (Glycine soja Sieb. and Zucc.) is unique in agricultural ecosystems owing to its ability to grow in saline-alkali soils and fix atmospheric nitrogen via symbiotic interactions with diverse soil microbes. However, this rhizosphere microbiome and the nodule endosymbionts have not been investigated to identify PGPR. In this study, we investigated the structural and functional rhizosphere microbial communities in saline-alkali soil from the Yellow River Delta and coastal soil in China, as well as wild soybean root nodule endosymbionts. To reveal the composition of the microbial ecosystem, we performed 16S rRNA and nifH gene amplicon sequencing on root nodules and root zones under different environmental conditions. In addition, we used culture-independent methods to examine the root bacterial microbiome of wild soybean. For functional characterization of individual members of the microbiome and their impact on plant growth, we inoculated isolates from the root microbiome with wild soybean and observed nodulation. Sinorhizobium/Ensifer accounted for 97% of the root nodule microbiome, with other enriched members belonging to the phyla Actinobacteria, Bacteroidetes, Chloroflexi, Acidobacteria, and Gemmatimonadetes; the genera Sphingomonas, Microbacterium, Arthrobacter, Nocardioides, Streptomyces, Flavobacterium, Flavisolibacter, and Pseudomonas; and the family Enterobacteriaceae. Compared to saline-alkali soil from the Yellow River Delta, coastal soil was highly enriched for soybean nodules and displayed significant differences in the abundance and diversity of β-proteobacteria, δ-proteobacteria, Actinobacteria, and Bacteroidetes. Overall, the wild soybean root nodule microbiome was dominated by nutrient-providing Sinorhizobium/Ensifer and was enriched for bacterial genera that may provide salt resistance. Thus, this reductionist experimental approach provides an avenue for future systematic and functional studies of the plant root microbiome.

Manufacturing industries generate much waste, which can be converted into raw materials for other allied industries. Thermoplastic wastes can be recycled in many cases. Recently these recycled plastics have been used to manufacture the input filament necessary for material extrusion additive manufacturing (MEAM). Polylactic acid (PLA) is one of the most common and potentially useful engineering plastics that can be additively manufactured, but the effect of recycling on the effective properties is still not well known. In this study, the mechanical properties of MEAM-processed PLA were compared for virgin and recycled feedstock. The collected data were analyzed using a full-factorial design of experiments method, providing insight into the effects of layer thickness and infill density on the performance of the material before and after recycling. The results showed a notable degradation in properties after recycling, but this effect was moderated by modifying the studied parameters. This work and its conclusions will serve as a screening study to guide future efforts in this area and promote the wider use of recycled materials in additive manufacturing.

The inventory model of seasonal products is developed in which the deterioration controlled by investing some cost of item preservation technology. In diminishing market, price sensitiveness analyzed with an exponentially declining demand. The object is to optimize the total profit per unit time, optimal selling price, length of replenishment cycle and preservation technology investment. We have shown that there exist an optimal selling price, and length of replenishment cycle that maximize the total profit. Also, we have proved the total profit per unit time is a concave function of the preservation technology investment. Finally, the model is illustrated by numerical example and fallowed by sensitive analysis.

In this work, we present an ecofriendly, non-hazardous, green synthesis of zinc oxide nanoparticles (ZnO NPs) by leaf extract of Crotalaria verrucosa (C. verrucosa). Total phenolic content, total flavonoid and total protein contents of C. verrucosa were determined. Further, synthesized ZnO NPs was characterized by UV–visible spectroscopy (UV-vis), X-ray diffractometer (XRD), Fourier transform infra-red (FTIR) Spectra, transmission electron microscope (TEM), and Dynamic light scattering (DLS) analysis. UV-vis shows peak at 375 nm which is unique to ZnO NPs. XRD analysis demonstrates the hexagonal phase structures of ZnO NPs. FTIR spectra demonstrates the molecules and bondings associated with the synthesized ZnO NPs and assures the role of phytochemical compounds of C. verrucosa in reduction and capping of ZnO NPs. TEM image exhibits that the prepared ZnO NPs is hexagonal shaped and in size ranged between 16 to 38 nm which is confirmed by DLS. Thermo-gravimetric analysis (TGA) was performed to determine the thermal stability of biosynthesized nanoparticles during calcination. The prepared ZnO NPs showed significant antibacterial potentiality against Gram-positive (S. aureus) and Gram-negative (Proteus vulgaris, Klebsiella pneumoniae, and Escherichia coli) pathogenic bacteria and SEM image shows the generalized mechanism of action in bacterial cell after NPs internalization. In addition, NPs are also found to be effective against the studied cancer cell lines for which cytotoxicity was assessed using MTT assay and results demonstrate highest growth of inhibition at the concentration of 100 µg/mL with IC50 value at 7.07 µg/mL for HeLa and 6.30 µg/mL for DU145 cell lines, in contrast to positive control (C. verrucosa leaf extract) with IC50 of 22.30 µg/mL on HeLa cells and 15.72 µg/mL on DU145 cells. Also, DAPI staining was performed in order to determine the effect on nuclear material due to ZnO NPs treatment in the studied cell lines taking leaf extract as positive control and untreated negative control for comparison. Cell migration assay was evaluated to determine the direct influence of NPs on metastasis that is potential suppression capacity of NPs to tumor cell migration. Outcome of the synthesized ZnO NPs using C. verrucosa shows antimicrobial activity against studied microbes, also cytotoxicity, apoptotic mediated DNA damage and antiproliferative potentiality in the studied carcinoma cells and hence, can be further used in biomedical, pharmaceutical and food processing industries as an effective antimicrobial and anti-cancerous agent.

Abstract: Dickeya sp., a plant pathogen, causing soft rot with strong pectin degradation capacity was taken for the comprehensive analysis of its corresponding biomass degradative system, which has not been analyzed yet. Whole genome sequence analysis of the isolated soft-rotten plant pathogen Dickeya sp. WS52, revealed various coding genes which involved in vegetable stalk degradation-related properties. A total of 122 genes were found to be encoded for putative carbohydrate-active enzymes (CAZy) in Dickeya sp. WS52. The number of pectin degradation-related genes, was higher than that of cellulolytic bacteria as well as other Dickeya spp. strains. The CAZy in Dickeya sp.WS52 contains a complete repertoire of enzymes required for hemicellulose degradation, especially pectinases. In addition, WS52 strain possessed plenty of genes encoding potential ligninolytic relevant enzymes, such as multicopper oxidase, catalase/hydroperoxidase, glutathione S-transferase, and quinone oxidoreductase. Transcriptome analysis revealed that parts of genes encoding lignocellulolytic enzymes were significantly upregulated in the presence of minimal salt medium with vegetable stalks. However, most of the genes were related to lignocellulolytic enzymes, especially pectate lyases and were downregulated due to the slow growth and downregulated secretion systems. The assay of lignocellulolytic enzymes including CMCase and pectinase activities were identified to be more active in vegetable stalk relative to MSM + glucose. However, compared with nutrient LB medium, it needed sufficient nutrient to promote growth and to improve the secretion system. Further identification of enzyme activities of Dickeya sp.WS52 by HPLC confirmed that monosaccharides were produced during degradation of tomato stalk. This identified degradative system is valuable for the application in the lignocellulosic bioenergy industry and animal production.

Two unrecognizable strains of the same bacterial species form a distinct colony boundary. During growth as colonies, Myxococcus xanthus uses multiple factors to establish cooperation between recognized strains and prevent interactions with unrecognized strains of the same species. Here, ΔMXAN_0049 is a mutant strain deficient in immunity for the paired nuclease gene, MXAN_0050, that has a function in the colony-merger incompatibility of Myxococcus xanthus DK1622. With the aim to investigate the factors involved in boundary formation, a proteome and metabolome study was employed. Visualization of the boundary between DK1622 and ΔMXAN_0049 was done scanning electron microscope (SEM), which displayed the presence of many damaged cells in the boundary. Proteome analysis of the DK1622- boundary disclosed many possible proteins, such as cold shock proteins, cell shape-determining protein MreC, along with a few pathways, such as RNA degradation, phenylalanine, tyrosine and tryptophan biosynthesis, and Type VI secretion system (T6SS), which may play major roles in the boundary formation. Metabolomics studies revealed various secondary metabolites that were significantly produced during boundary formation. Overall, the results concluded that multiple factors participated in the boundary formation in M. xanthus, leading to cellular damage that is helpful in solving the mystery of the boundary formation mechanism.

Myxococcus xanthus DK1622 is a rich source of novel secondary metabolites, and it is often used as an expression host of exogenous biosynthetic gene clusters. However, the frequency of obtaining large genome-deletion variants by using traditional strategies is low, and progenies generated by homologous recombination contain irregular deletions. The present study aims to develop an efficient genome-engineering system for this bacterium based on the Cre/loxP system. We first verified the functionality of the native cre system that was integrated into the chromosome with an inducible promoter PcuoA. Then we assayed the deletion frequency of 8-bp-spacer-sequence mutants in loxP by Cre recombinase which was expressed by suicide vector pBJ113 or self-replicative vector pZJY41. It was found that higher guanine content in a spacer sequence had higher deletion frequency, and the self-replicative vector was more suitable for the Cre/loxP system, probably due to the leaky expression of inducible promoter PcuoA. We also inspected the effects of different antibiotics and the native or synthetic cre gene. Polymerase chain reaction (PCR) and sequencing of new genome joints confirmed that the Cre/loxP system was able to delete a 466 kb fragment in M. xanthus. This Cre/loxP-mediated recombination could serve as an alternative genetic manipulation method.