{"title":"Synergistic anti-biofilm strategy based on essential oils and its application in the food industry.","authors":"Pei Yang, Yuxiao Huo, Qingli Yang, Fangyuan Zhao, Changjian Li, Jian Ju","doi":"10.1007/s11274-025-04289-8","DOIUrl":"10.1007/s11274-025-04289-8","url":null,"abstract":"<p><p>The microbial biofilm can induce a variety of food safety problems, and cause huge economic losses. Essential oils (EOs) not only have broad-spectrum antibacterial activity but also have a good ability to inhibit biofilm. However, the addition dose of EOs in practical application usually exceeds their flavor threshold, resulting in the appearance of undesired flavor. Therefore, synergistic antimicrobial may be a potential strategy to improve the antibacterial activity of EOs and to reduce their dosage. This paper focuses on the analysis of the synergistic anti-biofilm strategies based on EOs. Based on these, the action mechanism of EOs against biofilm and other commonly used anti-biofilm strategies in the food industry are summarized. The anti-biofilm mechanism of EOs is mainly related to inhibiting the synthesis of extracellular polysaccharides and proteins, destroying biofilm structure, inhibiting the metabolic activity of biofilm, inhibiting quorum sensing (QS) and regulating the formation of biofilm and the expression of toxicity-related genes. At present, the commonly used anti-biofilm strategies in the food industry mainly include physical strategies, chemical strategies and biological strategies, among which the combined application of different strategies is the future development trend. In particular, the synergistic anti-biofilm strategy based on EOs has shown great application value in the food industry. To sum up, some new information in this paper will give guidance and provide more reference for the development of efficient biofilm regulation strategies in future.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 3","pages":"81"},"PeriodicalIF":4.0,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143516886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Qing Li, Jihui Jiang, Haoyang Cheng, Bin Zhang, Yang Zhang, Jian Hu
{"title":"Changes in root-associated bacterial communities across growth stages of salt-tolerant and salt-sensitive rice grown in coastal saline-alkali soils.","authors":"Qing Li, Jihui Jiang, Haoyang Cheng, Bin Zhang, Yang Zhang, Jian Hu","doi":"10.1007/s11274-025-04284-z","DOIUrl":"10.1007/s11274-025-04284-z","url":null,"abstract":"<p><p>In this study, the root-associated bacterial communities of salt-tolerant and salt-sensitive rice grown in coastal saline-alkali soils were characterized at three major growth stages (jointing, heading and maturity) using Illumina MiSeq sequencing. The results showed that the growth stage had a stronger influence on endophytic bacterial diversity than the genotype, with diversity decreasing as growth progressed. However, the rhizospheric bacterial diversity was minimally affected by both growth stage and genotype. The variation in both rhizospheric and endophytic bacterial communities was primarily driven by the growth stages, but was also influenced by the genotype. Interestingly, there were distinct differences in changes in taxon abundance between the rhizospheric and endophytic bacterial communities, suggesting that the assembly mechanisms of these communities may differ, particularly in salt-sensitive rice. Additionally, certain genera were found to be enriched in the rhizosphere and endosphere compared to the bulk soil, and this enrichment varied depending on the growth stage and genotype. Notably, the dominant bacterium Clostridium was consistently enriched in the endosphere of salt-tolerant rice throughout all growth stages. This genus was also found to be more abundant overall in the rhizosphere and endosphere of salt-tolerant rice compared to salt-sensitive rice. These findings provide a deeper understanding of the co-evolution between rice and its associated microbes, and offer valuable insights for the isolation and application of beneficial root-associated bacteria in coastal saline-alkali soils.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 3","pages":"76"},"PeriodicalIF":4.0,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143516867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Identification and biodegradation characterization of high-density polyethylene using marine bacteria isolated from the coastal region of the Arabian Sea, at Gujarat, India.","authors":"Raj Kumar Sardar","doi":"10.1007/s11274-025-04283-0","DOIUrl":"10.1007/s11274-025-04283-0","url":null,"abstract":"<p><p>High-Density Polyethylene (HDPE) PE is one of the primary contributors of long-lasting and prolonged pollution in the environment. In this study, more than three hundred marine isolates collected off the Gujarat Sea coast were tested for HDPE plastic utilizing ability. Among fifty-one positive noted isolates, RS124 as a potential strain was identified as Micrococcus flavus (accession is PP858228) based on 16 S rRNA gene sequencing and total cellular fatty acid profiling. Initial bacterial adherence on the film surface was shown in a scanning electron microscopy (SEM) image as a key step to biodegradation. Moreover, atomic force microscopy (AFM) shows that the film surface became more fragile, damaged, and rougher than untreated films. Shifts and alterations in peak transmittance with emergence of two new shouldered peak in degraded HDPE observed by fourier transform infrared spectroscopy (FTIR) was associated to chemical and mechanical alteration. Thermogravimetric analysis (TGA) analysis designated larger difference in percent weight loss provisions thermal instability. In the enzymatic study, the highest activity of peroxidase and dehydrogenase was recorded on the 3rd and 4th weeks of treatment with strain, respectively, during co-incubation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis disclosed the presence of a distinct 19 kDa size protein, uncovering its role in the colonization of bacteria on the hydrophilic HDPE surfaces. About 1.8% weight reduction in HDPE was recorded as a result after 30 days of bio-treatment with M. flavus. Hence, the entire observed results reveal that the M. flavus RS124 could be effectively applied for the degradation of HDPE. This is the first report on M. flavus that it exhibits plastic degrading characteristic ever, which may allow for green scavenging of plastic waste.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 3","pages":"74"},"PeriodicalIF":4.0,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143516868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jamile Nazari, Vahide Payamnoor, Mohammad Reza Kavosi, Jahanbakhsh Asadi
{"title":"Boosting triterpene production in cell suspension culrures of Betula pendula Roth: the role of endophytic and basidiomycete fungi isolated from birch trees.","authors":"Jamile Nazari, Vahide Payamnoor, Mohammad Reza Kavosi, Jahanbakhsh Asadi","doi":"10.1007/s11274-025-04293-y","DOIUrl":"10.1007/s11274-025-04293-y","url":null,"abstract":"<p><p>This research aims to improve the synthesis of triterpenoids in cell suspension cultures of Betula pendula Roth stem cells using biological elicitors, including basidiomycete fungi (Arthrinium arundinis, Hyphodontia paradoxa, and Stereum hirsutum) and endophytes (Fusarium fuckelii, Alternaria alternata, and Penicillium citrinum). Five of the mentioned fungi were isolated from the bark, while A. arundinis was isolated from lichen of birch trees. Their extracellular extracts were used as elicitors. In most cases, these factors were significantly higher than the control. The growth index of Birch stem cells under the influence of different elicitors mostly yielded positive results. The highest percentage of cell viability was observed after a 24-hour treatment with 300 mg/L of F. fuckelii extract. Elicitation for 24 h with P. citrinum at concentrations of 150 and 450 mg/L, respectively, caused the greatest increase in betulin (8.8 times) and its terpenoid derivatives (2 times) compared to the control. A 36-hour elicitation with H. paradoxa at 450 mg/L resulted in a fourfold increase in betulinic acid compared to the control. The highest function of betulin and betulinic acid was achieved with A. alternata at concentrations of 300 mg/L and 150 mg/L respectively, within 36 h of exposure time. The highest function of terpenoid derivatives was achieved after 12 h of elicitation with 450 mg/L P. citrinum (5 times). This report presents, the positive effect of fungal elicitors extracted from birch trees on induced betulin, betulinic acid, and their triterpenoid derivatives production in cell suspension cultures of Betula pendula, for the first time.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 3","pages":"86"},"PeriodicalIF":4.0,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143516864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sara A Sedeek, Fedja Farowski, Stella Youssafi, Anastasia Tsakmaklis, Susanne Brodesser, Madiha M El-Attar, Mohamed Omar Abdelmalek, Maria J G T Vehreschild
{"title":"In vitro validation concept for lyophilized fecal microbiota products with a focus on bacterial viability.","authors":"Sara A Sedeek, Fedja Farowski, Stella Youssafi, Anastasia Tsakmaklis, Susanne Brodesser, Madiha M El-Attar, Mohamed Omar Abdelmalek, Maria J G T Vehreschild","doi":"10.1007/s11274-025-04291-0","DOIUrl":"10.1007/s11274-025-04291-0","url":null,"abstract":"<p><p>Fecal microbiota transplantation (FMT) effectively treats recurrent Clostridioides difficile infection (rCDI), typically administered as a fresh or frozen stool suspension through colonoscopy, nasojejunal tube, or oral capsules. Lyophilized fecal microbiota (LFM) are an alternative to frozen FM products. We aimed to assess whether lyophilization affects bacterial viability and metabolite levels and to develop LFM capsules for clinical use in Germany. Fecal donations from pre-screened volunteers were aliquoted and analyzed through microbial cell counting, bacterial culture, 16S rRNA gene amplicon sequencing, and bile acid assays. Results showed higher counts of viable bacterial cells and cultured anaerobes in unprocessed stool compared to freshly processed stool (p = 0.012 and p < 0.001, respectively). No significant difference in viable bacterial counts was found between freshly processed (day 0), lyophilized (day 3) and frozen FM (day 3) (p = 0.15), nor between freshly processed (day 0), lyophilized (days 30 and 90) and frozen FM (day 30) (p = 0.07). lyophilization did not significantly impact bile acid and 16S rRNA profiling. Encapsulation of lyophilized powder required fewer capsules (10-14) than frozen capsules (30). LFM products are a practical, viable alternative to frozen and fresh FM products, potentially improving storage and patient acceptance.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 3","pages":"83"},"PeriodicalIF":4.0,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11865215/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143516870","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
E S Lisintta, Annie George, Sona Gafoor, S Vishnupriya, Jesmi Yousuf, Bhavya Kachiprath, A K Valsa, Ally C Antony, P A Aneesa, A A Mohamed Hatha
{"title":"A psychrotolerant extracellular phosphatase from Krossfjorden sediment bacterium Bacillus cereus KR_O9: purification and functional characterization.","authors":"E S Lisintta, Annie George, Sona Gafoor, S Vishnupriya, Jesmi Yousuf, Bhavya Kachiprath, A K Valsa, Ally C Antony, P A Aneesa, A A Mohamed Hatha","doi":"10.1007/s11274-025-04252-7","DOIUrl":"10.1007/s11274-025-04252-7","url":null,"abstract":"<p><p>This study reports the identification of a phosphatase-producing psychrotolerant bacterium, Bacillus cereus KR_O9, isolated from Arctic Krossfjorden sediments. The extracellular KR_O9 phosphatase was purified to homogeneity through acetone precipitation and ion exchange chromatography, achieving a specific activity of 12.51 U mg⁻<sup>1</sup> min⁻<sup>1</sup>, a 17.3-fold purification, and a 64.94% yield. The molecular mass of the enzyme was estimated as 37 kDa by SDS-PAGE. K<sub>m</sub>, and V<sub>max</sub> for KR_O9 phosphatase towards substrate p-NPP were determined as 0.033 M L<sup>-1</sup>, 1.67 mg<sup>-1</sup> min<sup>-1</sup>. The enzyme exhibited optimal activity at 40 °C and pH 6.0, with stability up to ≤ 40 °C and at pH levels ≤ 7.0, highlighting its thermolability and sensitivity to alkaline pH. At 10 mM level, Cu<sup>2+</sup>, Hg<sup>2+</sup>, Co<sup>2+</sup>, Pb<sup>2+</sup>, urea, EDTA, and GSH significantly stimulated the enzyme, and Ca<sup>2+</sup>, Zn<sup>2+</sup>, Mn<sup>2+</sup>, SDS, and CTAB were, strong inhibitors. The kinetic efficiency, resilience to low to neutral pH, and tolerance to various metal ions, including heavy metals, underscore the potential of B. cereus KR_O9 phosphatase for bioremediation and biofertilizer applications, paving the way for industrial exploration.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 2","pages":"72"},"PeriodicalIF":4.0,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143415126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Indu Pant, Akhilesh A Potnis, Ravindranath Shashidhar
{"title":"Gene knockout studies of Dps protein reveals a novel role for DNA-binding protein in maintaining outer membrane permeability.","authors":"Indu Pant, Akhilesh A Potnis, Ravindranath Shashidhar","doi":"10.1007/s11274-025-04269-y","DOIUrl":"10.1007/s11274-025-04269-y","url":null,"abstract":"<p><p>DNA-binding proteins like Dps are crucial for bacterial stress physiology. This study investigated the unexpected role of Dps protein in maintaining outer membrane integrity of Salmonella Typhimurium. We observed that a Δdps mutant displayed increased sensitivity to glycopeptide antibiotics (vancomycin, nisin), which are ineffective against Gram-negative bacteria due to their thick outer membrane (OM). Furthermore, the Δdps mutant exhibited susceptibility to membrane-disrupting agents like detergents (deoxycholate, SDS) and phages. The perforation was observed in OM after the treatment of vancomycin using atomic force microscopy. Notably, this sensitivity was rescued by supplementing the media with calcium and magnesium cations. These findings suggest a novel function for Dps in maintaining outer membrane permeability. We propose two potential mechanisms: 1) Dps might directly localize to the outer membrane 2) Dps might regulate genes responsible for lipopolysaccharide synthesis or outer membrane proteins, key components of outer membrane. This study highlights a previously unknown role for Dps beyond DNA binding and warrants further investigation into the precise mechanism by which it influences outer membrane integrity in Salmonella.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 2","pages":"70"},"PeriodicalIF":4.0,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11821673/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143411125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Xenosiderophores: bridging the gap in microbial iron acquisition strategies.","authors":"Ravinsh Kumar, Ashutosh Singh, Amrita Srivastava","doi":"10.1007/s11274-025-04287-w","DOIUrl":"10.1007/s11274-025-04287-w","url":null,"abstract":"<p><p>Microorganisms acquire iron from surrounding environment through specific iron chelators known as siderophores that can be of self-origin or synthesized by neighboring microbes. The latter are termed as xenosiderophores. The acquired iron supports their growth, survival, and pathogenesis. Various microorganisms possess the ability to utilize xenosiderophores, a mechanism popularly termed as 'siderophore piracy' besides synthesizing their own siderophores. This adaptability allows microorganisms to conserve energy by reducing the load of siderogenesis. Owing to the presence of xenosiderophore transport machinery, these microbial systems can be used for targeting antibiotics-siderophore conjugates to control pathogenesis and combat antimicrobial resistance. This review outlines the significance of xenosiderophore utilization for growth, stress management and virulence. Siderogenesis and the molecular mechanism of its uptake by related organisms have been discussed vividly. It focuses on potential applications like disease diagnostics, drug delivery, and combating antibiotic resistance. In brief, this review highlights the importance of xenosiderophores projecting them beyond their role as mere iron chelators.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 2","pages":"69"},"PeriodicalIF":4.0,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143411130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Victor U Chigozie, Morteza Saki, Charles O Esimone
{"title":"Molecular structural arrangement in quorum sensing and bacterial metabolic production.","authors":"Victor U Chigozie, Morteza Saki, Charles O Esimone","doi":"10.1007/s11274-025-04280-3","DOIUrl":"10.1007/s11274-025-04280-3","url":null,"abstract":"<p><p>Quorum sensing (QS) regulates bacterial behaviors such as biofilm formation, virulence, and metabolite production through signaling molecules like acyl-homoserine lactones (AHLs), peptides, and AI-2. These signals are pivotal in bacterial communication, influencing pathogenicity and industrial applications. This review explores the molecular architecture of QS signals and their role in metabolite production, emphasizing structural modifications that disrupt bacterial communication to control virulence and enhance industrial processes. Key findings highlight the development of synthetic QS analogs, engineered inhibitors, and microbial consortia as innovative tools in biotechnology and medicine. The review underscores the potential of molecular engineering in managing microbial behaviors and optimizing applications like biofuel production, bioplastics, and anti-virulence therapies. Additionally, cross-species signaling mechanisms, particularly involving AI-2, reveal new opportunities for regulating interspecies cooperation and competition. This synthesis aims to bridge molecular insights with practical applications, showcasing how QS-based technologies can drive advancements in microbial biotechnology and therapeutic strategies.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 2","pages":"71"},"PeriodicalIF":4.0,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143411128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Lactobacillomics as a new notion in lactic acid bacteria research through omics integration.","authors":"Özge Kahraman Ilıkkan","doi":"10.1007/s11274-025-04285-y","DOIUrl":"10.1007/s11274-025-04285-y","url":null,"abstract":"<p><p>Omics technologies are a set of disciplines that analyze large-scale molecular data to understand biological systems in a holistic way. These technologies aim to reveal the structure, functions and interactions of organisms by studying processes at many levels of biomolecules, from the genome to metabolism. Lactobacillomics is introduced as an interdisciplinary field that integrates multiple \"omics\" technologies-including genomics, transcriptomics, proteomics, metabolomics, and metagenomics- to provide a comprehensive insight into \"lactic acid bacteria\" species. Lactobacillomics aims to elucidate the genetic, metabolic, and functional characteristics of lactic acid bacteria (LAB) species, providing insights into the mechanisms underlying their probiotic effects and contributions to the host microbiome. By analyzing genomes and metabolic pathways, researchers can identify specific genes responsible for health-promoting functions and desirable fermentation characteristics, which can guide the development of targeted probiotic strains with optimized health benefits. The integration of these omics data allows facilitating the discovery of biomarkers for health and disease states, the development of new probiotics tailored to specific populations or health conditions, and the optimization of fermentation processes to enhance the safety, flavor, and nutritional profile of fermented foods. A comprehensive review and bibliometric analysis were conducted to provide an overview of this promising field between 2005 and 2025 by examining Web of Science Core Collection data. Research results reveal trending topics, future perspectives, and key areas of growth within lactic acid bacteria (LAB) studies, particularly as they intersect with omics technologies.</p>","PeriodicalId":23703,"journal":{"name":"World journal of microbiology & biotechnology","volume":"41 2","pages":"68"},"PeriodicalIF":4.0,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11811450/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143392108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}