Bioresource Technology Reports最新文献

{"title":"Optimizing PHB recovery from Cupriavidus necator under mild acidic and alkaline digestion conditions","authors":"Darlington Nnabodo ,&nbsp;Emmanuel Olusola Oke ,&nbsp;Mariusz Gackowski ,&nbsp;Joanna Kryściak-Czerwenka ,&nbsp;Taghi Miri ,&nbsp;Matthew Keith ,&nbsp;Robert Karcz ,&nbsp;Maciej Guzik","doi":"10.1016/j.biteb.2025.102323","DOIUrl":"10.1016/j.biteb.2025.102323","url":null,"abstract":"<div><div>Polyhydroxybutyrate (PHB), a biopolymer produced by microbes, has gained attention as a sustainable alternative to traditional fossil-based plastics. Although solvent extraction is commonly used for PHB recovery, its environmental and health risks underline the need for safer and greener methods. Acidic and alkaline digestion techniques have gained interest as affordable options, but most reported processes rely on harsh chemical and thermal conditions that can degrade the polymer and hinder scalability. This study applies Response Surface Methodology (RSM) with Central Composite Design (CCD) to optimize PHB recovery under mild digestion conditions using NaOH (0.02–0.2 M, 1–5 h, 30–70 °C) and H₂SO₄ (0.02–0.5 M, 1–5 h at 50 °C). For <em>Cupriavidus necator</em> H16 (∼40 wt% PHB), optimal acidic digestion (0.5 M H₂SO₄, 2 h) yielded ∼92 % recovery and 60 % purity, while optimized alkaline digestion (0.02 M NaOH, 1 h, 55 °C) achieved ∼82 % recovery and 75 % purity. Notably, higher PHB content (&gt;65 wt%) under mild alkaline conditions enabled complete recovery (100 %) and &gt; 97 % purity, with the same protocol also effective for <em>Zobelella denitrificans</em>. Compared to chloroform extraction (∼41 % recovery, ∼69 % purity), the mild digestion protocols substantially improved both yield and environmental profile. Structural and thermal characterization (FTIR, ¹H/¹³C MAS NMR, GPC, TG-DSC) confirmed minimal polymer degradation, highlighting the potential of these mild, scalable methods for biomedical and packaging applications.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102323"},"PeriodicalIF":0.0,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145321600","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":"Isolation of microcrystalline cellulose from vegetable waste biomass and its potential as a high-performance green reinforcement in biocomposites","authors":"Indran Suyambulingam ,&nbsp;Raja Somasundaram ,&nbsp;Divya Divakaran ,&nbsp;Senthilkumar Boominathan ,&nbsp;P. Senthamaraikannan ,&nbsp;Nadir Ayrilmis ,&nbsp;Murni Handayani","doi":"10.1016/j.biteb.2025.102337","DOIUrl":"10.1016/j.biteb.2025.102337","url":null,"abstract":"<div><div>Global problems on waste disposal and sustainable material production require an innovative concept for recycling and utilization of waste materials. This work was centred toward isolation and comprehensively characterizing of microcrystalline cellulose from a market waste. High purity microcrystalline cellulose was obtained through a chemical process that uses tetraethylenetetramine, tartaric acid, sulfamic acid and sodium percarbonate in sequence. The extracted microcrystalline cellulose had a yield of 38 %, density of 1.491 g/cm³ and an impressive cellulose content of 95.19 %. Characterization analysis such as Fourier-transform infrared spectroscopy, X-ray diffractometer, ultraviolet-visible spectroscopy, thermal analysis, and morphological analysis showed that the synthesized material was highly crystalline with crystallinity index of 73.8 % and average particle size of 103.696 μm. The microcrystalline cellulose had a good thermal stability with a thermal degradation temperature of 228.56 °C and energy of activation of 74.16 kJ/mol. The scanning electron microscopy analysis revealed a rough and porous surface with distinct crystal bundles and average particle size of 103.696 μm which were beneficial in improving interfacial adhesion in composites. Additional surface roughness parameters determined by atomic force microscopy scans yielded negative skewness values, and higher kurtosis to support the material's applicability for mechanical reinforcement. The findings revealed that the microcrystalline cellulose derived from agricultural waste disposal was a sustainable and effective reinforcement for advanced composites. In addition, the microcrystalline cellulose supports the circular bioeconomy and achievement of sustainable development goal (SDG 12) established by the United Nations in 2015, which focuses on responsible consumption and production.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102337"},"PeriodicalIF":0.0,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145321584","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":"Recirculating aquaculture systems: Advances, impacts, and integrated pathways for sustainable growth","authors":"Zubair Hashmi ,&nbsp;Faizah Metali ,&nbsp;Muhamad Amin ,&nbsp;Muhammad Saifullah Abu Bakar ,&nbsp;Yusuf Wibisono ,&nbsp;Wahyunanto Agung Nugroho ,&nbsp;Muhammad Roil Bilad","doi":"10.1016/j.biteb.2025.102340","DOIUrl":"10.1016/j.biteb.2025.102340","url":null,"abstract":"<div><div>Recirculating aquaculture systems (RAS) offer a transformative approach to sustainable fish and shrimp farming by minimizing water use, enabling high-density production, and providing precise environmental control. Between 2020 and 2025, innovations such as low-head oxygenation, modular biofilters, integrated microalgae units, AI-driven feeding, and cold atmospheric plasma disinfection improved efficiency, biosecurity, and sustainability. This review synthesizes advances in engineering design, water quality management, automation, species-specific welfare optimization, and economic performance. Integrating microalgae and microbial consortia enhances nutrient recycling, CO₂ sequestration, oxygen production, and biofilter resilience, advancing zero-waste objectives. Concurrently, optimized flow regimes, lighting, and feeding protocols improve growth performance and welfare across species. Despite progress, challenges in energy intensity, microbial stability, system complexity, and cost scalability remain. Future directions include renewable energy integration, standardized protocols, interdisciplinary training, and supportive policies to accelerate adoption of circular, precision aquaculture. This review provides a critical resource in designing resilient, next-generation RAS.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102340"},"PeriodicalIF":0.0,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145321576","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":"Advanced photobioreactor strategy for chromium phycoremediation with Spirulina platensis","authors":"Erisson Paulo Borges Lopes,&nbsp;Larissa Nayhara Soares Santana Falleiros,&nbsp;Fabiana Regina Xavier Batista","doi":"10.1016/j.biteb.2025.102343","DOIUrl":"10.1016/j.biteb.2025.102343","url":null,"abstract":"<div><div>The study investigates the phycoremediation of Cr(VI) by <em>Spirulina platensis</em> under static cultivation (batch system) and a tubular photobioreactor. An inoculum of <em>Spirulina platensis</em> (1.2 g L⁻¹) was exposed to 30 and 60 mg L⁻¹ Cr(VI). Cellular resistance was verified until day seven, after which chromium exposure led to cell growth inhibition and reduced chlorophyll production. Chromium removal was verified through two mechanisms: initial passive biosorption on cell walls, facilitated by functional groups identified via Fourier-transform infrared spectroscopy, followed by active intracellular bioaccumulation, where Cr(VI) was reduced to less toxic Cr(III) via enzymatic activity. Scanning electron microscopy and energy-dispersive X-ray spectroscopy confirmed the adsorption of chromium on the cell surface. In the static system, final biomass reached 0.65 g L⁻¹ (46 % reduction), while the photobioreactor produced 3.19 g L⁻¹ (18.2 % below control). Total chromium removal efficiencies reached 69.3 % (10 days) and 67.1 % (7 days) in static and photobioreactor systems, respectively.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102343"},"PeriodicalIF":0.0,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145321583","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":"Extraction of polysaccharides from waste medium as functional ingredients","authors":"Nan Zhao,&nbsp;Zhongyang Liu,&nbsp;Xinyue Fan,&nbsp;Muhammad Naveed Sheas,&nbsp;Xinyi Chen,&nbsp;Ting Yu,&nbsp;Fujie Yan","doi":"10.1016/j.biteb.2025.102334","DOIUrl":"10.1016/j.biteb.2025.102334","url":null,"abstract":"<div><div>The post-fermentation culture medium, an underutilized resource, contains substantial amounts of bioactive polysaccharides. To overcome the drawbacks of conventional extraction techniques such as high energy consumption, environmental pollution, and structural degradation, this study adopted alcohol precipitation, a green method offering mild conditions, solvent reusability, and preserved bioactivity. A total polysaccharide yield of 3.75 g/L was obtained from the waste medium. Following purification via ion-exchange chromatography, the polysaccharide fraction PS1 demonstrated significant biological properties. It exhibited antibacterial effects in zebrafish infected with <em>Staphylococcus aureus</em> and <em>Pseudomonas fluorescens</em>, inhibited biofilm formation by 40.86 % and 29.04 % at 1 mg/mL, respectively, and displayed antioxidant capacity. PS1 consisted of mannose, <em>N</em>-acetylglucosamine, glucose confirmed by HPLC and its structure was further characterized by FT-IR, and NMR analysis. As the first report on the extraction and bioactivity assessment of polysaccharides from this waste source, our work offers a sustainable strategy for recycling post-fermentation residues and advancing resource conservation.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102334"},"PeriodicalIF":0.0,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263509","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":"Optimizing potential of consortium AKUC-1 for cresol bioremediation: A statistical approach complemented with AI-based prediction","authors":"Apurva Kadia,&nbsp;Urvish Chhaya","doi":"10.1016/j.biteb.2025.102342","DOIUrl":"10.1016/j.biteb.2025.102342","url":null,"abstract":"<div><div>Biodegradation is an economically feasible, environmentally friendly, and sustainable method for removing organic pollutants from environmental matrices. This work explores the potential of consortium AKUC-1 for bioremediation of cresol isomer mixtures (ortho, meta-, and para-cresol). Microbiome profiling of the consortium revealed dominance of the phylum Firmicutes and the genus <em>Bacillus</em>. To identify key parameters and improve the degradation efficiency of cresol isomers, AKUC-1 was statistically optimized using Response Surface Methodology — Central Composite Design (RSM-CCD), with support from a machine learning tool, support vector machine (SVM). Analysis of Variance confirmed the accuracy of the RSM-CCD model, with a significant F-value of 56.76 and a <em>p</em>-value of less than 0.0001, indicating the model's robustness. Under optimal conditions, with 3.50 g L⁻¹ sucrose, 0.0075 g L⁻¹ FeCl₃, and 0.0075 g L⁻¹ CaCl₂, the consortium degraded 82.54 % of 600 ppm cresol isomers, which is 2.58 times higher than in unoptimized media.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102342"},"PeriodicalIF":0.0,"publicationDate":"2025-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263510","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":"Quality of black soldier fly (Hermetia illucens L.) larvae frass according to feeding substrates in Dschang (West Cameroon)","authors":"Tchoumi Gabriele Rodine Ngassa ,&nbsp;Agoume Fatimatou Agada ,&nbsp;Ottou Francine Mba ,&nbsp;Danwe Rigobert Haiwa ,&nbsp;Mbougue Estelle Voukeng ,&nbsp;Rousseau Djouaka ,&nbsp;Aoudou Yaouba ,&nbsp;Dieudonné Bitom","doi":"10.1016/j.biteb.2025.102332","DOIUrl":"10.1016/j.biteb.2025.102332","url":null,"abstract":"<div><div>The global population growth not only requires a significant increase in food production but also leads to an increase in waste production especially in livestock farming. Facing this challenge, innovative waste transformation techniques for sustainable agriculture have emerged, such as the bioconversion of waste by insect larvae, particularly the black soldier fly (<em>Hermetia illucens</em> L.). Black soldier fly larvae (BSFL) are gaining attention as a valuable protein source in animal feed while the residues, frass, are used as organic fertilizer. However its properties are not well understood. The work aimed to characterize the feeding substrates and the corresponding frass focusing on their chemical and microbiological properties. The animal droppings used as substrates, and the resulting frass were sampled and analyzed. Chemical analyses were performed using spectrophotometry. Fungi and bacteria were identified based on their morphological and cultural characteristics. The results showed that pig manure produced the biggest larvae and the greatest quantity of frass. Compared to the respective feeding substrates, the frass contained higher levels of organic matter, organic carbon, total nitrogen, phosphorus and potassium. Treatment with BSFL also reduced the heavy metal contents in the manure. Microbiological analysis revealed fungi from the genera <em>Cladosporium</em>, <em>Aspergillus</em> and <em>Phoma</em> except in the pig manure-derived frass. Bacteria identified included species <em>Actinomycetes</em>, <em>Clostridium</em>, <em>Staphylococcus</em>, <em>Bacillus</em> and <em>Yersinia</em>. Notably, BSFL eliminated <em>Actinomycetes</em> in all frass samples. These findings highlight the potential of BSFL as an effective solution for waste management and frass production.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102332"},"PeriodicalIF":0.0,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263432","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":"Microbial nanofactories: A mini-review on sustainable biosynthesis of nanomaterials","authors":"Chen Lyu ,&nbsp;Xiaomin Li ,&nbsp;Yanan Yin ,&nbsp;Jian Yang ,&nbsp;Chongbin Zhang ,&nbsp;Zeping Zhang ,&nbsp;Cheng Wang ,&nbsp;Jianlong Wang","doi":"10.1016/j.biteb.2025.102322","DOIUrl":"10.1016/j.biteb.2025.102322","url":null,"abstract":"<div><div>Microorganisms, as emerging biosynthetic platforms, have shown significant potential in materials science, catalytic conversion, and biomedical fields. Their inherent morphologies and surface active groups can mediate the synthesis of special structured nanomaterials such as multi-level porous microspheres and high specific surface area nanorods that significantly optimize the physical and chemical properties of the materials. This efficient, economical, and sustainable synthesis paradigm provides an innovative approach for the precise preparation of nanomaterials. This article provides a systematic review of the microorganisms used in nanomaterials synthesis, which were extensively introduced, and types of nanomaterials synthesis utilizing microorganisms were systematically described. Approaches to nanomaterials synthesis utilizing microorganisms were elaborated, and the applications of microbially synthesized nanomaterials were comprehensively presented.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102322"},"PeriodicalIF":0.0,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263433","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":"From waste to wonder: The potential of protein hydrolysates as plant biostimulants in agriculture","authors":"Padmapriya Dhanasekaran ,&nbsp;Shanthi Chittibabu ,&nbsp;Said Mouzeyar ,&nbsp;Aurélia Boulaflous-Stevens ,&nbsp;Cedric Delattre ,&nbsp;Jane Roche","doi":"10.1016/j.biteb.2025.102333","DOIUrl":"10.1016/j.biteb.2025.102333","url":null,"abstract":"<div><div>Protein hydrolysates (PHs) derived from plant and animal byproducts have emerged as promising biostimulants in sustainable agriculture. This comprehensive review investigates the production, composition, and mechanisms of action of PHs in promoting plant growth and stress resilience. PHs are obtained through enzymatic hydrolysis of protein-rich waste, yielding mixtures of amino acids and bioactive peptides. Commercial PHs exhibit diverse effects, including enhanced nutrient uptake, abiotic stress tolerance, and crop yield in various agricultural and horticultural species. The biostimulant activity of PHs is attributed to multiple interconnected pathways, such as hormone-like signalling, antioxidant defense, osmotic regulation, and gene expression modulation. Moreover, PHs influence the plant microbiome, potentially contributing to improved stress adaptation. Foliar and root application methods, along with factors like pH and transporters, govern PH absorption and translocation in plants. Transcriptomic and proteomic studies have provided insights into the molecular mechanisms underlying PH-mediated growth promotion and stress mitigation. Future research should focus on optimizing PH formulations, exploring synergies with other biostimulants, and elucidating the role of plant-microbe interactions. By harnessing the potential of these sustainable compounds, PH biostimulants offer a promising approach to enhance agricultural productivity and resilience in the face of global challenges.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102333"},"PeriodicalIF":0.0,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145321578","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":"Advanced treatment of sugarcane vinasse via hybrid photocatalytic sequencing batch biofilm reactor (HP-SBBR): A combined physicochemical and biological approach","authors":"Wei-Chin Kee ,&nbsp;Yee-Shian Wong ,&nbsp;Soon-Jian Low ,&nbsp;Soon-An Ong ,&nbsp;Nabilah Aminah Lutpi ,&nbsp;Sumate Chaiprapat ,&nbsp;Santhana Krishnan ,&nbsp;Audrey Chai ,&nbsp;Boon-Beng Lee","doi":"10.1016/j.biteb.2025.102335","DOIUrl":"10.1016/j.biteb.2025.102335","url":null,"abstract":"<div><div>A large volume of undesired chemical liquid generated by ethanol distillation is known as sugarcane vinasse. It is an acidic, dark brown distillery spent wash rich in organic compounds that can cause toxicity to living organisms, soil and water acidification, and groundwater contamination. This study introduces a hybrid chemical and biological treatment approach combining photocatalysis and a sequencing batch biofilm reactor (HP-SBBR). Chemical coagulation with alum was applied as a pretreatment due to its compatibility and low cost. Subsequently, the bio-photocatalytic reaction in HP-SBBR with zinc oxide (ZnO) photocatalyst achieved substantial degradation, evidenced by 93.1 % COD reduction and 99.7 % decolourisation at a low loading rate of 0.024 kg·COD/m³·day. Kinetic studies and UV–vis spectra confirmed that the combination of light and microorganisms effectively reduce COD of sugarcane vinasse and improved the biodegradation index. Microbial analysis using 16S rDNA revealed predominant bacterial genera, including <em>Pseudomonas</em> sp. (28 %), <em>Bacillus</em> sp. (8 %), <em>Tissierella</em> sp. (7 %), and <em>Azoarcus</em> sp. (6 %). To assess toxicity, a phytotoxicity test using mung bean (<em>Vigna radiata</em>) showed an 85 % reduction in toxicity based on germination rate, indicating that treated vinasse is more suitable for reuse in agriculture. Although the integrated pre-coagulation and HP-SBBR system demonstrated strong technical feasibility and treatment performance, further studies on operating costs and technical constraints are necessary to evaluate its applicability and impact in full-scale vinasse management.</div></div>","PeriodicalId":8947,"journal":{"name":"Bioresource Technology Reports","volume":"32 ","pages":"Article 102335"},"PeriodicalIF":0.0,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263430","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}