Biocatalysis and agricultural biotechnology最新文献

{"title":"Biopriming of Solanum lycopersicum seeds with novel root endophytic bacterial consortium retrieved from halotolerant Sundarban mangroves to sustain growth and yield with salt resilience","authors":"Anwesha Chatterjee ,&nbsp;Proma Ghosh ,&nbsp;Murali Sharaff ,&nbsp;Prateek Madhab Bhattacharya ,&nbsp;Harshata Pal","doi":"10.1016/j.bcab.2024.103414","DOIUrl":"10.1016/j.bcab.2024.103414","url":null,"abstract":"<div><div>Mangroves are often found in coastal areas and tropical wetlands that can withstand high salinity. We hypothesized that endophytes that are harbouring in the roots of mangrove plants may improve the innate immunity of host plants to survive naturally in saline environment. Retrieving these endophytes and sequential characterization may function as a novel bio-effector for non-host food crops as well. We focused on the integrated approach towards formulating a novel bacterial consortium. Thirty-one bacterial endophytes isolated from the roots of mangrove plants were screened for plant growth promoting potential by inoculating our model crop (Tomato). Seven most promising isolates impacting plant growth were identified. <em>In-vitro</em> plant growth promoting characters were also analysed. The root colonization by the isolates was confirmed by Scanning Electron Microscopy (SEM). Among the screened isolates, four of them were found to be compatible with each other when grown together and were selected to formulate a novel biostimulant consortia. The consortia treated Tomato plants exhibited superior phenological characters. In the pot experiment, plant height of the treated plants was about ≈43 cm while the non-treated plants under salt stress could grow only up to a height of ≈26 cm. Similarly, a total fruit yield of ≈6.8 kg was observed in case of treated plants under salt stress whereas the non-treated plants under salt stress could only produce ≈4.7 kg of fruit. This study demonstrated that the beneficial bacteria inhabiting in mangrove roots can increase the potential of conferring salt tolerance to non-host crops, thereby contributing to sustainable agriculture.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142553479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Accumulation of podophyllotoxin in the root culture of Hyptis suaveolens (L.) Poit: A potential natural lignan for clinically useful anticancer drugs","authors":"Sâmia Torres Silva ,&nbsp;Rafael Marlon Alves de Assis ,&nbsp;Adriane Duarte Coelho ,&nbsp;Simony Carvalho Mendonça ,&nbsp;Lurdeslaine Faria Texeira ,&nbsp;Suzan Kelly Vilela Bertolucci ,&nbsp;José Eduardo Brasil Pereira Pinto","doi":"10.1016/j.bcab.2024.103416","DOIUrl":"10.1016/j.bcab.2024.103416","url":null,"abstract":"<div><div>Podophyllotoxin (PTOX) is a natural antiviral, antirheumatic and anticancer molecule, but it is expensive to chemically synthesize. The present study aimed to evaluate growth and PTOX accumulation in root cultures of <em>Hyptis suaveolens</em> treated with different concentrations of auxins, vitamins and myo-inositol. Root cultures grown in liquid medium supplemented with different concentrations of indole-3-butyric acid (IBA), 1-naphthaleneacetic acid (NAA), vitamins and myo-inositol were used to measure biomass production and PTOX content. PTOX quantitation was performed via high-performance liquid chromatography with diode-array detection (HPLC-DAD) using a newly developed analytical method after successful validation. Root culture in MS medium supplemented with 1 mg L⁻¹ IBA +0.5 mg L⁻¹ NAA resulted in the highest root dry weight (248.76 mg) and the highest PTOX concentration in the roots (179.97 μg g⁻¹ root). The roots cultured in liquid MS medium supplemented with 1 mg L⁻¹ IBA +0.5 mg L⁻¹ NAA presented the greatest increase in root biomass and PTOX content. This adequate balance of vitamin and myo-inositol supplementation in liquid MS culture medium increased the root dry weight and PTOX content.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142528556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microalgae to remove pharmaceutical and personal care products (PPCPs) from wastewater","authors":"Madhumita Ravikumar ,&nbsp;Karrun Velmurugan ,&nbsp;Ashwini J. John ,&nbsp;Ethiraj Selvarajan","doi":"10.1016/j.bcab.2024.103415","DOIUrl":"10.1016/j.bcab.2024.103415","url":null,"abstract":"<div><div>Substances used for medicinal, cosmetic, hygiene, and health care objectives are included in the category of pharmaceuticals and personal care products (PPCPs). They are a major source of public concern because of their ubiquitous manufacturing, usage, and careless disposal of expired products into the environment. Many PPCPs, including antibiotics, analgesics, endocrine disruptors, and microbial/disinfecting agents, are commonly detected in freshwater systems, groundwater, and wastewater treatment effluents in amounts ranging from nanograms per liter to milligrams per milliliter. Additionally, these compounds frequently show persistence and accumulate in biological tissue, eventually finding their way into crops, vegetables, and drinking water supplies. Because many PPCPs are known to have the ability to upset ecosystems and/or provide health hazards, they are categorized as “emerging contaminants.” The research on the occurrence, fate, and possible health and environmental hazards related to PPCPs in both aquatic and terrestrial habitats is thoroughly reviewed in this work. It also covers reported cases of danger or health concerns in humans, although a full assessment may not be possible given the limitations of the data at hand. This review aims to provide a comprehensive and well-focused overview of the current understanding of bioremediation strategies for the removal of pharmaceuticals and personal care products (PPCPs), with an emphasis on the application of macroalgae, microalgae, and aquatic macrophytes. It examines the physicochemical properties of PPCPs and their potential risks to environmental and human health. Additionally, the review explores the potential and challenges associated with the broader application of biotechnologies employing algae and aquatic macrophytes. This includes research efforts to correlate the operational parameters of these biotechnologies with the primary mechanisms responsible for PPCP removal. In conclusion, algae and macrophytes present promising, eco-friendly solutions for wastewater treatment, significantly contributing to the mitigation of PPCP contamination.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142528557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bio-prospecting fluoride tolerant bacteria for their optimistic contribution in instigating resilience against fluoride stress in Oryza sativa L.","authors":"Priya Katiyar ,&nbsp;Neha Pandey ,&nbsp;S. Keshavkant","doi":"10.1016/j.bcab.2024.103412","DOIUrl":"10.1016/j.bcab.2024.103412","url":null,"abstract":"<div><div>In recent years, the effects of fluoride (F) pollution in numerous ecosystems such as groundwater, soil, <em>etc</em>. Have become a major issue worldwide. This increase in F pollution is a direct consequence of the unbridled use of fertilizers in agricultural and several other human activities that require immediate and appropriate action. Therefore, this manuscript reveals important findings on the efficacy of bacteria isolated from agricultural fields in central Chhattisgarh in manifesting resistance to F and in reversing the F-induced oxidative damage in susceptible <em>Oryza sativa</em> L, (Var. MTU1010). Chronic exposure of <em>Oryza sativa</em> L. to sodium fluoride (NaF) (50 mg L⁻¹) severely impeded growth and various physiological parameters such as germination percentage, biomass and root and shoot length and stimulated the formation of reactive oxygen species (ROS), which enhanced electrolyte leakage and formation of cytotoxic products like malondialdehyde. To this end, potential bacterial strains, namely MT2A, MT3A, MT4A, and Du3A were isolated, screened for various plant growth promoting (PGP) traits and used to explore their efficiency to mitigate F toxicity in <em>Oryza sativa</em> L. <em>in vivo.</em> The seedlings inoculated with the bacterial strains showed significant development as evidenced by an increase in root and shoot length, biomass and chlorophyll content. Additionally, inoculation of these strains in combination with F stress significantly decreased oxidative stress by increasing the expression of protective genes encoding antioxidant enzymes and boosted agronomic traits remarkably. Overall, the manuscript demonstrates the pivotal role played by the isolated bacteria in abating ill effects of F in the <em>Oryza sativa</em> L. seedlings and proves their potential as protective bioagents against F stress.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142553477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “Green integrative large scale treatment of tannery effluent, CO2 sequestration, and biofuel production using oleaginous green microalga Nannochloropsis oculata TSD05: An ecotechnological approach Title of article” [Biocatal. Agric. Biotechnol. 61 (2024) 103370]","authors":"Silambarasan Tamil Selvan ,&nbsp;Arul Balasubaramanian ,&nbsp;BalaKumaran Manickam Dakshinamoorthi ,&nbsp;Balasubramanian Velramar ,&nbsp;Mohammad Fareed","doi":"10.1016/j.bcab.2024.103410","DOIUrl":"10.1016/j.bcab.2024.103410","url":null,"abstract":"","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142528558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Co-metabolism of substrates by Pseudomonas aeruginosa NG4 regulates biosurfactant production","authors":"Shivangi Sankhyan ,&nbsp;Prasun Kumar ,&nbsp;Madan Sonkar ,&nbsp;Sandeep Kumar ,&nbsp;Soumya Pandit ,&nbsp;Subhasree Ray","doi":"10.1016/j.bcab.2024.103382","DOIUrl":"10.1016/j.bcab.2024.103382","url":null,"abstract":"<div><div>As compared to chemical surfactants, biosurfactants exhibit a more significant impact in various sectors due to their degradability, stability, and other specific physicochemical properties. Biosurfactant production by <em>Pseudomonas aeruginosa</em> NG4 was studied by co-metabolism of 1% glucose (wv⁻¹) and 1% CG (vv⁻¹) with different time intervals. The addition of Glucose (0h) + CG (24 h) (1:1, wv⁻¹, vv⁻¹) to the culture medium resulted in 4.23 gL^-1 of biosurfactant production. An enhancement of about 4.75-fold in the yield was observed. The time period of incubation of CG in the feed was optimized to be 24 h after the initiation of fermentation with glucose. Among the several nitrogen sources used in this study, sodium nitrate produced more biosurfactant than the others. Biochemical characterization through FTIR and LC-MS of biosurfactant reflected the presence of mono- and di-rhamnolipids. Therefore, this study highlights the ability of <em>P. aeruginosa</em> NG4 to increase biosurfactant production by co-utilizing substrates.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142421012","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Essential oils from plant resources as potent insecticides and repellents: Current status and future perspectives","authors":"B. Jyotsna ,&nbsp;Swaroopa Patil ,&nbsp;Y. Surya Prakash ,&nbsp;P. Rathnagiri ,&nbsp;P.B. Kavi Kishor ,&nbsp;N. Jalaja","doi":"10.1016/j.bcab.2024.103395","DOIUrl":"10.1016/j.bcab.2024.103395","url":null,"abstract":"<div><div>The extensive use of chemical pesticides on crop plants is harmful to mankind as well as to insect predators. Their long-term usage contaminates water, soil, and the food chain with the accumulation of toxic residues. Plant-based essential oils with a broad spectrum of antifungal, larvicidal, ovicidal, and pesticidal activities are now being preferred as an alternative and sustainable source to safeguard crop plants. Evidence now exists that essential oils alter the nervous and locomotor activities of insects. They target specific and may affect the various systems of the insect pests. This may generally do no or less harm to mammals including humans when compared to chemical insect pests, therefore they can be deployed as relatively safer molecules to reign in the devastating pests and reduce economic losses. They are cost-effective, available throughout the year, non-persistent in the environment, and vertebrate or non-target toxicity is minimal in comparison with chemical pesticides. This brief review focuses on the current status of essential oils used as potent pesticides, stored-product insect pest reduction, insect repellents, and fungicides. This also suggests the way forward to design superior strategies such as the technique of nano-encapsulation to ameliorate the stability and stave off their volatilization.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142420569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bio-synthesized AGS@AgNPs for wound healing, antioxidant support, antibacterial defense, and anticancer intervention","authors":"Majid Zare-Bidaki ,&nbsp;Pouria Mohammadparast-Tabas ,&nbsp;Mahboobe Khorashadizade ,&nbsp;Peyman Mohammadparast-Tabas ,&nbsp;Esmat Alemzadeh ,&nbsp;Amirhossein Saberi ,&nbsp;Hamid Kabiri-Rad ,&nbsp;Samira Eghbali","doi":"10.1016/j.bcab.2024.103402","DOIUrl":"10.1016/j.bcab.2024.103402","url":null,"abstract":"<div><div>Slow wound healing and wound infections are significant challenges for burn patients. Biosynthesis of nanoparticles (NPs) using plant extracts offers a rapid, facile, and safe method for producing biocompatible NPs. In this study, <em>Anethum graveolens</em> seed (AGS) extract served as a masking and reducing agent for the synthesis of silver nanoparticles (AGS@AgNPs). Initially, the parameters influencing AGS@AgNPs synthesis, including silver nitrate concentration, reaction time, temperature, and pH, were optimized. Subsequently, AGS@AgNPs structural and biological characteristics were evaluated. Observation of the surface plasmon resonance (SPR) of the AGS@AgNPs at approximately 420 nm, accompanied by a color change of the suspension to dark brown, confirmed AGS@AgNPs synthesis. Analysis via X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS), zeta potential, and transmission electron microscopy (TEM) confirmed the production of pure, homogeneous, spherical, and stable silver NPs with sizes ranging from 20 to 40 nm using Anethum graveolens extract. Antibacterial assays revealed significant activity against both Gram-positive and Gram-negative bacterial strains. Antioxidant assessments demonstrated that AGS@AgNPs at a concentration of 250 μg/ml exhibited 92% inhibition of DPPH free radicals and in FRAP test, <em>A. graveolens</em> seed extract and AGS@AgNPs reduced Fe³⁺ ions to Fe²⁺ by 48.7% and 73.1%, respectively. Investigation of anticancer properties against the lung cancer cell line (A-549) revealed an IC₅₀ value of 242 μg/ml. The results of the flow cytometry test and lactate dehydrogenase (LDH) assay demonstrated the death of cancer cells. Moreover, wound healing assays demonstrated a significant acceleration in burn wound closure rates in rats on days 3, 7, and 14 following treatments with AGS@AgNPs. Overall, these findings highlight the favorable biological activities of AGS@AgNPs, suggesting their potential utility in nanomedicine applications.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142421331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phytopurifying activity of Salvia officinalis L. from Tunisia","authors":"Soumaya Khaldi ,&nbsp;Kaouther Mechergui ,&nbsp;Assila Ben Salem ,&nbsp;Wahbi Jaouadi","doi":"10.1016/j.bcab.2024.103404","DOIUrl":"10.1016/j.bcab.2024.103404","url":null,"abstract":"<div><div>The sage (<em>Salvia officinalis</em> L.) is an annual and biennial plant of Mediterranean origin from the Lamiaceae family. It's a valuable medicinal plant, which is widely used in traditional medicine in world and in Tunisia; this species is very rich in active biological compounds.</div><div>The objective of this study was to determine the phyto-purifying activity of sage essential, fixed oil and ethanolic extracts in an extensive wastewater purification system of Tunisian national sanitation office. We have measured the physico-chemical parameters of wastewater treated with sage oils and extracts. Then, we have carried out a bacteriological analysis of this wastewater.</div><div>Our results showed that pH values measured following treatment with sage essential oil gave a pH close to neutral. Therefore, this water can be used for irrigation. In addition, the sage leaves and stems have a considerable effect on the salinity and caused a very remarkable increase in electrical conductivity of the wastewater, the measure of elimination rate demonstrated the effectiveness of sage extracts in eliminating all the microorganisms existing in wastewater.</div><div>In fact, sage essential oil has the greatest effectiveness in reducing the chemical oxygen demand of the wastewater and we have noticed that a decrease in chemical oxygen demand of microorganisms existing in the wastewater was necessarily accompanied by a decrease in biological oxygen demand. Therefore, sage leaves and stems were more effective in reducing suspended matter existing in wastewater.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142433391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Innovative approaches to combating dental biofilms: Nanotechnology and its applications","authors":"Akankshya Dash,&nbsp;Chinnasamy Ragavendran","doi":"10.1016/j.bcab.2024.103406","DOIUrl":"10.1016/j.bcab.2024.103406","url":null,"abstract":"<div><div>Dental biofilms, impacting around 80% of the global population, are a significant issue causing serious dental disorders like periodontitis, which has links to autoimmune diseases such as rheumatoid arthritis. Traditional biofilm management methods, including mechanical removal and chemical agents, often fail to completely eradicate biofilms, allowing oral infections to persist. Nanotechnology offers innovative solutions for medication delivery and biofilm disruption, presenting new opportunities for antimicrobial surfaces and healthcare equipment development. This paper explores advancements in nanotechnology-based strategies to combat dental biofilms, primarily formed by <em>Lactobacillus</em> species, <em>Actinomyces</em> species, and <em>Streptococcus mutans</em>, and examines the composition and characteristics of dental biofilms. The green synthesis of nanoparticles using plant-based ingredients demonstrates significant antibacterial activity against biofilm-associated pathogens. Additionally, the paper investigates the potential of various metal oxide-based nanoparticles, such as titanium, zinc, and silver, in dental treatments to inhibit biofilm formation and enhance oral health. The stages of dental biofilm formation are outlined, highlighting the risks of untreated biofilms, including systemic health implications. This review underscores the promise of nanodrugs in preventing dental biofilms and improving the efficacy of dental procedures. It delves into cutting-edge nanotechnological approaches, emphasizing the novel use of nanoparticles like gold, silver, and antimicrobial peptides, and their unique mechanisms for disrupting biofilm formation and enhancing dental treatments. The review also covers recent advancements in nanotechnology, offering innovative and more effective solutions for targeted dental care strategies, potentially revolutionizing the approach to managing dental biofilms and promoting better oral health outcomes.</div></div>","PeriodicalId":8774,"journal":{"name":"Biocatalysis and agricultural biotechnology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142433453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}