Commenced in January 2007
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Edition: International
Paper Count: 352

Search results for: fermentation

112 Production of Bioethanol from Oil PalmTrunk by Cocktail Carbohydrases Enzyme Produced by Thermophilic Bacteria Isolated from Hot spring in West Sumatera, Indonesia

Authors: Yetti Marlida, Syukri Arif, Nadirman Haska


Recently, alcohol fuels have been produced on industrial scales by fermentation of sugars derived from wheat, corn, sugar beets, sugar cane etc. The enzymatic hydrolysis of cellulosic materials to produce fermentable sugars has an enormous potential in meeting global bioenergy demand through the biorefinery concept, since agri-food processes generate millions of tones of waste each year (Xeros and Christakopoulos 2009) such as sugar cane baggase , wheat straw, rice straw, corn cob, and oil palm trunk. In fact oil palm trunk is one of the most abundant lignocellulosic wastes by-products worldwide especially come from Malaysia, Indonesia and Nigeria and provides an alternative substrate to produce useful chemicals such as bioethanol. Usually, from the ages 3 years to 25 years, is the economical life of oil palm and after that, it is cut for replantation. The size of trunk usually is 15-18 meters in length and 46-60 centimeters in diameter. The trunk after cutting is agricultural waste causing problem in elimination but due to the trunk contains about 42% cellulose, 34.4%hemicellulose, 17.1% lignin and 7.3% other compounds,these agricultural wastes could make value added products (Pumiput, 2006).This research was production of bioethanol from oil palm trunk via saccharafication by cocktail carbohydrases enzymes. Enzymatic saccharification of acid treated oil palm trunk was carried out in reaction mixture containing 40 g treated oil palm trunk in 200 ml 0.1 M citrate buffer pH 4.8 with 500 unit/kg amylase for treatment A: Treatment B: Treatment A + 500 unit/kg cellulose; C: treatment B + 500 unit/kgg xylanase: D: treatment D + 500 unit/kg ligninase and E: OPT without treated + 500 unit/kg amylase + 500 unit/kg cellulose + 500 unit/kg xylanase + 500 unit/kg ligninase. The reaction mixture was incubated on a water bath rotary shaker adjusted to 600C and 75 rpm. The samples were withdraw at intervals 12 and 24, 36, 48,60, and 72 hr. For bioethanol production in biofermentor of 5L the hydrolysis product were inoculated a loop of Saccharomyces cerevisiae and then incubated at 34 0C under static conditions. Samples are withdraw after 12, 24, 36, 48 and 72 hr for bioethanol and residual glucose. The results of the enzymatic hidrolysis (Figure1) showed that the treatment B (OPT hydrolyzed with amylase and cellulase) have optimum condition for glucose production, where was both of enzymes can be degraded OPT perfectly. The same results also reported by Primarini et al., (2012) reported the optimum conditions the hydrolysis of OPT was at concentration of 25% (w /v) with 0.3% (w/v) amylase, 0.6% (w /v) glucoamylase and 4% (w/v) cellulase. In the Figure 2 showed that optimum bioethanol produced at 48 hr after incubation,if time increased the biothanol decreased. According Roukas (1996), a decrease in the concentration of ethanol occur at excess glucose as substrate and product inhibition effects. Substrate concentration is too high reduces the amount of dissolved oxygen, although in very small amounts, oxygen is still needed in the fermentation by Saccaromyces cerevisiae to keep life in high cell concentrations (Nowak 2000, Tao et al. 2005). The results of the research can be conluded that the optimum enzymatic hydrolysis occured when the OPT added with amylase and cellulase and optimum bioethanol produced at 48 hr incubation using Saccharomyses cerevicea whereas 18.08 % bioethanol produced from glucose conversion. This work was funded by Directorate General of Higher Education (DGHE), Ministry of Education and Culture, contract no.245/SP2H/DIT.LimtabMas/II/2013

Keywords: oil palm trunk, enzymatic hydrolysis, saccharification

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111 Preharvest and Postharvest Factors Influencing Resveratrol, Myricetin and Quercetin Content of Wine

Authors: Mariam Khomasuridze, Nino Chkhartishvili, Irma Chanturia


The influence of preharvest and postharvest factors on resveratrol, myricetin and quercetin content of wine was studied during the experiment. The content of cis and trans resveratrol, myricetin and quercetin were analyzed by HPLC. In frame of experiment, the various factors affecting on wine composition were researched: variety, climate, viticulture practices, grape maturity, harvesting methods and wine making techniques. The results have shown that varietal potential and amount of yield play the most important role in formation of antioxidant compounds. Based on achieved results, the usage of medium roast oak chips protects resveratrol, myricetin, and quercetin from coagulation and precipitation. Compared to the control samples, the wines, produced by addition of oak chips were approximately four times richer with these antioxidant compounds. The retention of resveratrol was lowered with 45 % in wines, producing in Qvevri by Georgian traditional technology without controlling temperature during fermentation. The opposite effects in case of myricetin, quercetin and total phenolics content were determined. Their concentrations were higher with 56-78%, then in the fermented tank at 22 -25 °C. As the result of the experiment, the optimal technology scheme of wine was worked out, reached by biologically active compounds: resveratrol, myricetin, and quercetin.

Keywords: resveratrol, miricetin, quercetin, wine

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110 Extraction of Inulin from Cichorium Intybus and Its Application as Fat Replacer in Yoghurt

Authors: Hafiz Khuram Wasim Aslam, Muhammad Saeed, Azam Shakeel, Muhammad Inam Ur Raheem, Moazzam Rafiq Khan, Muhammad Atif Randhawa


Inulin is significant ingredient used in food industry that functions technologically as a fat replacer often without compromising taste and texture. In this study inulin was extracted from the chicory roots and the effect of inulin addition as a fat replacer on the physiochemical, microbiological and sensory properties of non-fat yogurt was investigated. The supplementation of chicory inulin reduced the magnitude of firmness in comparison with non-inulin ¬supplemented non-fat yoghurt. Higher values of acidity were observed due to the more microbial fermentation in the inulin containing yogurt as compared to non-inulin yogurt and were in the range of 0.56 to 0.75 during storage days. Syneresis in control sample increased from 43.9% to 47.9% during the storage study. However inulin addition at different treatment enhanced syneresis from 44.5% to 47.6%. Inulin addition at various concentrations caused an increase in the TPC due to its probiotic effect. No effects of inuline addition on fat and protein contents were observed. Non-fat yoghurt supplemented with inulin demonstrated sensory behavior better than that of the control yoghurt. The most important effect of the addition of inulin to non-fat yoghurt is an increase in the sensory attributes appearance, body and texture, taste and mouth feel, overall acceptability. On an average, yoghurt supplemented with 1 to 2% inulin was better in overall acceptance as compared to control yoghurt.

Keywords: inulin, fat replacer, yoghurt, sensory evaluation, low fat

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109 Integrated Process Modelling of a Thermophilic Biogas Plant

Authors: Obiora E. Anisiji, Jeremiah L. Chukwuneke, Chinonso H. Achebe, Paul C. Okolie


This work developed a mathematical model of a biogas plant from a mechanistic point of view, for urban area clean energy requirement. It aimed at integrating thermodynamics; which deals with the direction in which a process occurs and Biochemical kinetics; which gives the understanding of the rates of biochemical reaction. The mathematical formulation of the proposed gas plant follows the fundamental principles of thermodynamics, and further analysis were accomplished to develop an algorithm for evaluating the plant performance preferably in terms of daily production capacity. In addition, the capacity of the plant is equally estimated for a given cycle of operation and presented in time histories. A nominal 1500m3 biogas plant was studied characteristically and its performance efficiency evaluated. It was observed that the rate of biogas production is essentially a function of enthalpy ratio, the reactor temperature, pH, substrate concentration, rate of degradation of the biomass, and the accumulation of matter in the system due to bacteria growth. The results of this study conform to a very large extent with reported empirical data of some existing plant and further model validations were conducted in line with classical records found in literature.

Keywords: anaerobic digestion, biogas plant, biogas production, bio-reactor, energy, fermentation, rate of production, temperature, therm

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108 Physicochemical, Heavy Metals Analysis of Some Multi-Floral Algerian Honeys

Authors: Assia Amri, Naima Layachi, Ali Ladjama


The characterization of some Algerian honey was carried out on the basis of their physico-chemical properties: moisture,hydroxy methyl furfural, diastase activity, pH,free, total and lactonic acidity, electrical conductivity, minerals and proline content. Studied samples are found to be low in moisture and therefore safe from fermentation, low in HMF level and high in diastase activity. Additionally the diastase activity and the HMF content are widely recognized parameters indicating the freshness of honey. Phenolic compounds present in honey are classified into two groups - simple phenols and polyphenols. The simple phenols in honey are various phenol acids, but polyphenols are various flavonoids and flavonides. The aim of our work was to determine antioxidant properties of various Algerian honey samples–the total phenol content, total flavonoids content, as well as honey anti radical activity.The quality of honey samples differs on account of various factors such as season, packaging and processing conditions, floral source, geographical origin and storage period. It is important that precautions should be taken to ensure standardization and rationalization of beekeeping techniques, manufacturing procedures and storing processes to improve honey quality.

Keywords: honey, physico-chemical characterization, phenolic coumpound, HMF, diastase activity

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107 Microbial Diversity of El-Baida Marsh: Setif, Algeria

Authors: H. Necef, A. Benayad


Fungi are becoming more and more important in our life. Therefore, as a start for the symposium on filamentous fungi in biotechnology a short survey of the role of fungi in biotechnology. Salin soils occupy about 7% of land area; they are characterized by unsuitable physical conditions for the growth of living organisms. However, researches showed that some microorganisms especially fungi are able to grow and adapt to such extreme conditions; it is due to their ability to develop different physiological mechanisms in their adaptation. This is the first study on the physiological and biological characteristics of El-Beida marsh. Nine soil samples were taken at different points in two steps, the first was in winter (low temperature), and the second was in summer (high temperature). The physicochemical analyses of the soil were conducted, then the isolation process was applied using two methods, direct method and dilution method (10-1, 10-2, 10-3, 10-4). Different species of fungi were identified belong to 21 genera in addition to 3 yeast species, Aspergillus showed the highest proportion by 43%, then Penicillium by 20% then Alternaria by 7%, in addition to various genera in different proportions. As for the sampling periods, it was observed that the spread of fungi in winter was higher than in summer with the proportion 75.47% and 24.53% respectively. Some halotolerant fungi have a biotechnological importance especially if the salinity of the medium is necessary for the fermentation, and if the halotolerance genes of the fungus will define, this will open the research to study and improve this property for the industrial important micro-organisms.

Keywords: salinity, identification, aspergillus oryzae, halotolerance, fungi

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106 Antimicrobial Peptide Produced by Lactococcus garvieae with a Broad Inhibition Spectrum

Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep


By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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105 Assessment of cellulase and xylanase Production by chryseobacterium sp. Isolated from Decaying Biomass in Alice, Eastern Cape, South Africa

Authors: A. Nkohla, U. Nwodo, L. V. Mabinya, A. I. Okoh


A potential source for low-cost production of value added products is the utilization of lignocellulosic materials. However, the huddle needing breaching would be the dismantlement of the complex lignocellulosic structure as to free sugar base therein. the current lignocellosic material treatment process is expensive and not eco-friendly hence, the advocacy for enzyme based technique which is both cheap and eco-friendly is highly imperative. Consequently, this study aimed at the screening of cellulose and xylan degrading bacterial strain isolated from decaying sawdust samples. This isolate showed high activity for cellulase and xylanase when grown on carboxymethyl cellulose and birtchwood xylan as the sole carbon source respectively. The 16S rDNA nucleotide sequence of the isolate showed 98% similarity with that of Chryseobacterium taichungense thus, it was identified as a Chryseobacterium sp. Optimum culture conditions for cellulase and xylanase production were medium pH 6, incubation temperature of 25 °C at 50 rpm and medium pH 6, incubation temperature of 25 °C at 150 rpm respectively. The high enzyme activity obtained from this bacterial strain portends it as a good candidate for industrial use in the degradation of complex biomass for value added products.

Keywords: lignocellulosic material, chryseobacterium sp., submerged fermentation, cellulase, xylanase

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104 An Antifungal Peptide from Actinobacteria (Streptomyces Sp. TKJ2): Isolation and Partial Characterization

Authors: Abdelaziz Messis, Azzeddine Bettache, Nawel Boucherba, Said Benallaoua, Mouloud Kecha


Actinobacteria are of special biotechnological interest since they are known to produce chemically diverse compounds with a wide range of biological activity. This distinct clade of Gram-positve bacteria include some of the key antibiotic producers and are also sources of several bioactive compounds, established commercially a newly filamentous bacteria was recovered from Tikjda forest soil (Algeria) for its high antifungal activity against various pathogenic and phytopathogenic fungi. The nucleotide sequence of the 16S rRNA gene (1454 pb) of Streptomyces sp. TKJ2 exhibited close similarity (99 %) with other Streptomyces16S rRNA genes. Antifungal metabolite production of Streptomyces sp TKJ2 was evaluated using six different fermentation media. The extracellular products contained potent antifungal agents. Antifungal protein produced by Streptomyces sp. TKJ2 on PCA medium has been purified by ammonium sulfate precipitation, SPE column chromatography and high-performance liquid chromatography in a reverse-phase column. The UV chromatograms of the active fractions obtained at 214 nm by NanoLC-ESI-MS/MS have different molecular weights. The F20 Peptidic fraction obtained from culture filtrat of Streptomyces sp. TKJ2 precipitated at 30% of ammonium sulfate was selected for analysis by infusion ESI-MS which yielded a singly charged ion mass of 437.17 Da.

Keywords: actinobacteria, antifungal protein, chromatography, Streptomyces

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103 Control of Spoilage Fungi by Lactobacilli

Authors: Laref Nora, Guessas Bettache


Lactic acid bacteria (LAB) have a major potential to be used in biopreservation methods because they are safe to consume (GRAS: generally regarded as safe) and they naturally occurring microflora of many foods. The preservative action of LAB is due to several antimicrobial metabolites, including lactic acid, acetic acid, hydrogen peroxide, bacteriocins, carbon dioxide, diacetyl, and reuterin. Several studies have focused on the antifungal activity compounds from natural sources for biopreservation in alternatives to chemical use. LAB has an antifungal activity which may inhibit food spoilage fungi. Lactobacillus strains isolated from silage prepared in our laboratory by fermentation of grass in anaerobic condition were screened for antifungal activity with overlay assay against Aspergillus spp. The antifungal compounds were originated from organic acids; inhibitory activity did not change after treatment with proteolytic enzymes. Lactobacillus strains were able also to inhibit Trichoderma spp, Penicillium spp, Fusarium roseum, and Stemphylim spp by confrontation assay. The inhibitory activity could be detected against the mould Aspergillus spp in the apricot juice but not in a bakery product. These antifungal compounds have the potential to be used as food biopreservation to inhibit conidia germination, and mycelia growth of spoilage fungi depending on food type, pH of food especially in heat, and cold processed foods.

Keywords: lactic acid bacteria, Lactobacillus, Aspergillus, antifungal activity

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102 A Review of Antimicrobial Strategy for Cotton Textile

Authors: C. W. Kan, Y. L. Lam


Cotton textile has large specific surfaces with good adhesion and water-storage properties which provide conditions for the growth and settlement of biological organisms. In addition, the soil, dust and solutes from sweat can also be the sources of nutrients for microorganisms [236]. Generally speaking, algae can grow on textiles under very moist conditions, providing nutrients for fungi and bacteria growth. Fungi cause multiple problems to textiles including discolouration, coloured stains and fibre damage. Bacteria can damage fibre and cause unpleasant odours with a slick and slimy feel. In addition, microbes can disrupt the manufacturing processes such as textile dyeing, printing and finishing operations through the reduction of viscosity, fermentation and mold formation. Therefore, a large demand exists for the anti-microbially finished textiles capable of avoiding or limiting microbial fibre degradation or bio fouling, bacterial incidence, odour generation and spreading or transfer of pathogens. In this review, the main strategy for cotton textile will be reviewed. In the beginning, the classification of bacteria and germs which are commonly found with cotton textiles will be introduced. The chemistry of antimicrobial finishing will be discussed. In addition, the types of antimicrobial treatment will be summarized. Finally, the application and evaluation of antimicrobial treatment on cotton textile will be discussed.

Keywords: antimicrobial, cotton, textile, review

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101 Regulation of SHP-2 Activity by Small Molecules for the Treatment of T Cell-Mediated Diseases

Authors: Qiang Xu, Xingxin Wu, Wenjie Guo, Xingqi Wang, Yang Sun, Renxiang Tan


The phosphatase SHP-2 is known to exert regulatory activities on cytokine receptor signaling and the dysregulation of SHP-2 has been implicated in the pathogenesis of a variety of diseases. Here we report several small molecule regulators of SHP-2 for the treatment of T cell-mediated diseases. The new cyclodepsipeptide trichomides A, isolated from the fermentation products of Trichothecium roseum, increased the phosphorylation of SHP-2 in activated T cells, and ameliorated contact dermatitis in mice. The trichomides A’s effects were significantly reversed by using the SHP-2-specific inhibitor PHPS1 or T cell-conditional SHP-2 knockout mice. Another compound is a cerebroside Fusaruside isolated from the endophytic fungus Fusarium sp. IFB-121. Fusaruside also triggered the tyrosine phosphorylation of SHP-2, which provided a possible mean of selectively targeting STAT1 for the treatment of Th1 cell-mediated inflammation and led to the discovery of the non-phosphatase-like function of SHP-2. Namely, the Fusaruside-activated pY-SHP-2 selectively sequestrated the cytosolic STAT1 to prevent its recruitment to IFN-R, which contributed to the improvement of experimental colitis in mice. Blocking the pY-SHP-2-STAT1 interaction, with SHP-2 inhibitor NSC-87877 or using T cells from conditional SHP-2 knockout mice, reversed the effects of fusaruside. Furthermore, the fusaruside’s effect is independent of the phosphatase activity of SHP-2, demonstrating a novel role for SHP-2 in regulating STAT1 signaling and Th1-type immune responses.

Keywords: SHP-2, small molecules, T cell, T cell-mediated diseases

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100 Extractive Bioconversion of Polyhydroxyalkanoates (PHAs) from Ralstonia Eutropha Via Aqueous Two-Phase System-An Integrated Approach

Authors: Y. K. Leong, J. C. W. Lan, H. S. Loh, P. L. Show


Being biodegradable, non-toxic, renewable and have similar or better properties as commercial plastics, polyhydroxy alkanoates (PHAs) can be a potential game changer in the polymer industry. PHAs are the biodegradable polymer produced by bacteria, which are in interest as a sustainable alternative to petrochemical-derived plastics; however, its commercial value has significantly limited by high production and recovery cost of PHA. Aqueous two-phase system (ATPS) offers different chemical and physical environments, which contains about 80-90% water delivers an excellent environment for partitioning of cells, cell organelles and biologically active substances. Extractive bioconversion via ATPS allows the integration of PHA upstream fermentation and downstream purification process, which reduces production steps and time, thus lead to cost reduction. The ability of Ralstonia eutropha to grow under different ATPS conditions was investigated for its potential to be used in a bioconversion system. Changes in tie-line length (TLL) and a volume ratio (Vr) were shown to have an effect on PHA partition coefficient. High PHA recovery yield of 65% with a relatively high purity of 73% was obtained in PEG 6000/Sodium sulphate system with 42.6 wt/wt % TLL and 1.25 Vr. Extractive bioconversion via ATPS is an attractive approach for the combination of PHA production and recovery process.

Keywords: aqueous two-phase system, extractive bioconversion, polyhydroxy alkanoates, purification

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99 Performance of an Improved Fluidized System for Processing Green Tea

Authors: Nickson Kipng’etich Lang’at, Thomas Thoruwa, John Abraham, John Wanyoko


Green tea is made from the top two leaves and buds of a shrub, Camellia sinensis, of the family Theaceae and the order Theales. The green tea leaves are picked and immediately sent to be dried or steamed to prevent fermentation. Fluid bed drying technique is a common drying method used in drying green tea because of its ease in design and construction and fluidization of fine tea particles. Major problems in this method are significant loss of chemical content of the leaf and green appearance of tea, retention of high moisture content in the leaves and bed channeling and defluidization. The energy associated with the drying technology has been shown to be a vital factor in determining the quality of green tea. As part of the implementation, prototype dryer was built that facilitated sequence of operations involving steaming, cooling, pre-drying and final drying. The major findings of the project were in terms of quality characteristics of tea leaves and energy consumption during processing. The optimal design achieved a moisture content of 4.2 ± 0.84%. With the optimum drying temperature of 100 ºC, the specific energy consumption was 1697.8 kj.Kg-1 and evaporation rate of 4.272 x 10-4 Kg.m-2.s-1. The energy consumption in a fluidized system can be further reduced by focusing on energy saving designs.

Keywords: evaporation rate, fluid bed dryer, maceration, specific energy consumption

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98 Effect of Lignocellulose-Degrading Bacteria Isolated from Termite Gut on the Nutritive Value of Wheat Straw as Ruminant Feed

Authors: Ayoub Azizi-Shotorkhoft, Tahereh Mohammadabadi, Hosein Motamedi, Morteza Chaji, Hasan Fazaeli


This study was conducted to investigate nutritive value of wheat straw processed with termite gut symbiotic bacteria with lignocellulosic-degrading potential including Bacillus licheniformis, Ochrobactrum intermedium and Microbacterium paludicola in vitro. These bacteria were isolated by culturing termite guts contents in different culture media containing different lignin and lignocellulosic materials that had been prepared from water-extracted sawdust and wheat straw. Results showed that incubating wheat straw with all of three isolated bacteria increased (P<0.05) acid-precipitable polymeric lignin (APPL) compared to control, and highest amount of APPL observed following treatment with B. licheniformis. Highest and lowest (P<0.05) in vitro gas production and ruminal organic matter digestibility were obtained when treating wheat straw with B. licheniformis and control, respectively. However, other fermentation parameters such as b (i.e., gas production from the insoluble fermentable fractions at 144h), c (i.e., rate of gas production during incubation), ruminal dry matter digestibility, metabolizable energy, partitioning factor, pH and ammonia nitrogen concentration were similar between experimental treatments (P>0.05). It is concluded that processing wheat straw with isolated bacteria improved its nutritive value as ruminants feed.

Keywords: termite gut bacteria, wheat straw, nutritive value, ruminant

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97 Heterologous Expression of Heat-Shock Protein Improves Butanol Yield in a High-Speedy Growing Clostridium acetobutylicum Mutant

Authors: Min-Shiuan Liou, Yi Shan Yang, Yang-Zhan Huang, Chia-Wen Hsieh


A high speed growing and butanol-tolerant Clostridium acetobutylicum HOL1 mutant was screened throughout continuous adaption culture with C. acetobutylicum ATCC 824. The HOL1 strain can grow well in 10 g/L butanol contained CGM medium and can produce about 12.8 g /L butanol during 24 hrs. The C. acetobutylicum HOL1 strain was able to produce 166 mM butanol with 21 mM acetone at pH 4.8, resulting in a butanol selectivity (a molar ratio of butanol to total solvents) of 0.79, which is much higher than that (0.6) of the wild-type strain C. acetobutylicum ATCC 824. The acetate and butyrate accumulation were not observed during fermentation of the HOL1 strain. A hyper-butanol producing C. acetobutylicum HOL1 (pBPHS-3), which was created to overexpress the Bacillus psychrosaccharolyticus originated specific heat-shock protein gene, hspX, from a clostridial phosphotransbutyrylase promoter, was studied for its potential to produce a high titer of butanol. Overexpression of hspX resulted in increased final butanol yield 47% and 30% higher than those of the the ATCC824 and the HOL1 strains, respectively. The remarkable high-speed growth and butanol tolerance of strain HOL1 (pBPHS-3) demonstrates that overexpression of heterogeneous stress protein-encoding gene, hspX, could help C. acetobutylicum to effectively produce a high concentration of butanol.

Keywords: Clostridium acetobutylicum, butanol, heat-shock protein, resistance

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96 Authenticity of Ecuadorian Commercial Honeys

Authors: Elisabetta Schievano, Valentina Zuccato, Claudia Finotello, Patricia Vit


Control of honey frauds is needed in Ecuador to protect bee keepers and consumers because simple syrups and new syrups with eucalyptus are sold as genuine honeys. Authenticity of Ecuadorian commercial honeys was tested with a vortex emulsion consisting on one volume of honey:water (1:1) dilution, and two volumes of diethyl ether. This method allows a separation of phases in one minute to discriminate genuine honeys that form three phase and fake honeys that form two phases; 34 of the 42 honeys analyzed from five provinces of Ecuador were genuine. This was confirmed with 1H NMR spectra of honey dilutions in deuterated water with an enhanced aminoacid region with signals for proline, phenylalanine and tyrosine. Classic quality indicators were also tested with this method (sugars, HMF), indicators of fermentation (ethanol, acetic acid), and residues of citric acid used in the syrup manufacture. One of the honeys gave a false positive for genuine, being an admixture of genuine honey with added syrup, evident for the high sucrose. Sensory analysis was the final confirmation to recognize the honey groups studied here, namely honey produced in combs by Apis mellifera, fake honey, and honey produced in cerumen pots by Geotrigona, Melipona, and Scaptotrigona. This is a valuable contribution to protect honey consumers, and to develop the beekeeping industry in Ecuador.

Keywords: fake, genuine, honey, 1H NMR, Ecuador

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95 Using Human-Digestive Simulator to Harbor Encapsulated Lactobacillus casei 01 along with Pasteurized-Purple-Rice Drinks for Examination of the Health-Promoting Effects

Authors: Srivilai Worametrachanon, Arunee Apichartsrangkoon, Jiranat Techarang, Boonrak Phanchaisri


A human-digestive simulator consisted of four colon compartments, i.e., stomach, small intestine, proximal colon and distal colon used to harbor L. casei 01 plus either pasteurized ordinary-purple-rice drinks or germinated-purple-rice drinks. Accordingly, three treatment compositions had been set up and the effects of treatments on colon bacterial communities including their by-products were thoroughly examined. L. casei 01 plus purple-rice drinks gave rise to significantly high formation (P ≤ 0.05) of short-chain-fatty acids (SCFA) of which highest acetic acid was found followed by propionic and butyric acids, while the germinated-rice drink showed the greatest impact. Moreover, the effect was more pronounced upon prolonged fermentation. In addition, the influence of treatments on colon microbes was also demonstrated. Accordingly, desirable bacteria including colon Lactobacilli and Bifidobacteria were significantly increased (P ≤ 0.05) in both colons in comparison with the control and the effect was more prominent after adding purple-rice drink. On the other hand, undesirable Clostridia and coliforms were apparently diminished by the influence of treatment conditions, in which both compartments exhibited similar results.

Keywords: human-digestive simulator, Lactobacillus casei 01, Pasteurized-purple-rice drinks

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94 Device for Thermal Depolymerisation of Organic Substrates Prior to Methane Fermentation

Authors: Marcin Dębowski, Mirosław Krzemieniewski, Marcin Zieliński


This publication presents a device designed to depolymerise and structurally change organic substrate, for use in agricultural biogas plants or sewage treatment plants. The presented device consists of a heated tank equipped with an inlet valve for the crude substrate and an outlet valve for the treated substrate. The system also includes a gas conduit, which is at its tip equipped with a high-pressure solenoid valve and a vacuum relief solenoid valve. A conduit behind the high-pressure solenoid valve connects to the vacuum tank equipped with the outlet valve. The substrate introduced into the device is exposed to agents such as high temperature and cavitation produced by abrupt, short-term reduction of pressure within the heated tank. The combined effect of these processes is substrate destruction rate increase of about 20% when compared to using high temperature alone, and about 30% when compared to utilizing only cavitation. Energy consumption is greatly reduced, as the pressure increase is generated by heating the substrate. Thus, there is a 18% reduction of energy consumption when compared to a device designed to destroy substrate through high temperature alone, and a 35% reduction if compared to using cavitation as the only means of destruction.

Keywords: thermal depolymerisation, organic substrate, biogas, pre-treatment

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93 Enhancement in Digester Efficiency and Numerical Analysis for Optimal Design Parameters of Biogas Plant Using Design of Experiment Approach

Authors: Rajneesh, Priyanka Singh


Biomass resources have been one of the main energy sources for mankind since the dawn of civilization. There is a vast scope to convert these energy sources into biogas which is a clean, low carbon technology for efficient management and conversion of fermentable organic wastes into a cheap and versatile fuel and bio/organic manure. Thus, in order to enhance the performance of anaerobic digester, an optimizing analysis of resultant parameters (organic dry matter (oDM) content, methane percentage, and biogas yield) has been done for a plug flow anaerobic digester having mesophilic conditions (20-40°C) with the wet fermentation process. Based on the analysis, correlations for oDM, methane percentage, and biogas yield are derived using multiple regression analysis. A statistical model is developed to correlate the operating variables using the design of experiment approach by selecting central composite design (CCD) of a response surface methodology. Results shown in the paper indicates that as the operating temperature increases the efficiency of digester gets improved provided that the pH and hydraulic retention time (HRT) remains constant. Working in an optimized range of carbon-nitrogen ratio for the plug flow digester, the output parameters show a positive change with the variation of dry matter content (DM).

Keywords: biogas, digester efficiency, design of experiment, plug flow digester

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92 Effect of Nanoparticles Concentration, pH and Agitation on Bioethanol Production by Saccharomyces cerevisiae BY4743: An Optimization Study

Authors: Adeyemi Isaac Sanusi, Gueguim E. B. Kana


Nanoparticles have received attention of the scientific community due to their biotechnological potentials. They exhibit advantageous size, shape and concentration-dependent catalytic, stabilizing, immunoassays and immobilization properties. This study investigates the impact of metallic oxide nanoparticles (NPs) on ethanol production by Saccharomyces cerevisiae BY4743. Nine different nanoparticles were synthesized using precipitation method and microwave treatment. The nanoparticles synthesized were characterized by Fourier Transform Infra-Red spectroscopy (FTIR), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Fermentation processes were carried out at varied NPs concentrations (0 – 0.08 wt%). Highest ethanol concentrations were achieved after 24 h using Cobalt NPs (5.07 g/l), Copper NPs (4.86 g/l) and Manganese NPs (4.74 g/l) at 0.01 wt% NPs concentrations, which represent 13%, 8.7% and 5.4% increase respectively over the control (4.47 g/l). The lowest ethanol concentration (0.17 g/l) was obtained when 0.08 wt% of Silver NPs was used. And lower ethanol concentrations were observed at higher NPs concentration. Ethanol concentration decrease after 24 h for all the processes. In all set up with NPs, the pH was observed to be stable and the stability was directly proportional to nanoparticles concentrations. These findings suggest that the presence of some of the NPs in the bioprocesses has catalytic and pH stabilizing potential. Ethanol production by Saccharomyces cerevisiae BY4743 was enhanced in the presence of Cobalt NPs, Copper NPs and Manganese NPs. Optimization study using response surface methodology (RSM) will further elucidate the impact of these nanoparticles on bioethanol production.

Keywords: agitation, bioethanol, nanoparticles concentration, optimization, pH value

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91 Establishing a Microbial Co-Culture for Production of Cellulases Using Banana (Musa Paradisiaca) Pseudostem

Authors: Mulanga Luscious Mulaudzi, Ignatious Ncube


In nature, enzymatic degradation of lignocellulose is more efficient compared to in vivo bioprocessing. Thus, a co-culture should enable production of more efficient enzyme preparations that would mimic the natural decomposition of lignocellulose. The aim of the study was to establish a microbial co-culture for the production of highly active cellulase preparations. The objectives were to determine the use of a variety of culture media to isolate cellulose degrading microorganisms from decomposing banana pseudo stem and to optimize production of cellulase by co-cultures of microorganisms producing high levels of cellulose. Screening of fungal isolates was done on carboxylmethylcellulose agar plates which were stained with Congo red to show hydrolytic activity of the isolates. Co-culture and mixed culture of these microorganisms were cultured using Mendel salts with Avicel as the carbon source. Cultures were incubated at 30 °C with shaking at 200 rpm for 240 hrs. Enzyme activity assays were performed to determine endoglycosidase and β-glucosidase. Mixed culture of fungi-dead bacterial cells showed to be the best co-culture/ mixed culture to produce higher levels of cellulase activity in submerged fermentations (SmF) using Avicel™ as a carbon source. The study concludes use microorganism 5A in co-cultures is highly recommended in order to produce high amounts of β-glucosidases, no matter the combination used.

Keywords: avicel, co-culture, submerged fermentation, pseudostem

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90 Production of Novel Antibiotics by Importing eryK and eryG Genes in Streptomyces fradiae

Authors: Neda Gegar Goshe, Hossein Rassi


The antibacterial properties of macrolide antibiotics (such as erythromycin and tylosin) depend ultimately on the glycosylation of otherwise inactive polyketide lactones. Among the sugars commonly found in such macrolides are various 6-deoxyhexoses including the 3-dimethylamino sugars mycaminose and desosamine (4-deoxymycaminose). Some macrolides (such as tylosin) possess multiple sugar moieties, whereas others (such as erythromycin) have two sugar substituents. Streptomyces fradiae is an ideal host for development of generic polyketide-overproducing strains because it contains three of the most common precursors-malonyl-CoA, methylmalonyl-CoA and ethylmalonyl-CoA-used by modular PKS, and is a host that is amenable to genetic manipulation. As patterns of glycosylation markedly influence a macrolide's drug activity, there is considerable interest in the possibility of using combinatorial biosynthesis to generate new pairings of polyketide lactones with sugars, especially 6-deoxyhexoses. Here, we report a successful attempt to alter the aminodeoxyhexose-biosynthetic capacity of Streptomyces fradiae (a producer of tylosin) by importing genes from the erythromycin producer Saccharopolyspora erythraea. The biotransformation of erythromycin-D into the desired major component erythromycin-A involves two final enzymatic reactions, EryK-catalyzed hydroxylation at the C-12 position of the aglycone and EryG-catalyzed O methylation at the C-3 position of macrose .This engineered S. fradiae produced substantial amounts of two potentially useful macrolides that had not previously been obtained by fermentation.

Keywords: Streptomyces fradiae, eryK and eryG genes, tylosin, antibiotics

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89 Effects of Certain Natural Food Additives (Pectin, Gelatin and Whey Proteins) on the Qualities of Fermented Milk

Authors: Abderrahim Cheriguene, Fatiha Arioui


The experimental study focuses on the extraction of pectin, whey protein and gelatin, and the study of their functional properties. Microbiological, physicochemical and sensory approach integrated has been implanted to study the effect of the incorporation of these natural food additives in the matrix of a fermented milk type set yogurt, to study the stability of the product during the periods of fermentation and post-acidification over a period of 21 days at 4°C. Pectin was extracted in hot HCl solution. Thermo-precipitation was carried out to obtain the whey proteins while the gelatin was extracted by hydrolysis of the collagen from bovine ossein. The fermented milk was prepared by varying the concentration of the incorporated additives. The measures and controls carried performed periodically on fermented milk experimental tests were carried out: pH, acidity, viscosity, the enumeration of Streptococcus thermophilus, cohesiveness, adhesiveness, taste, aftertaste, whey exudation, and odor. It appears that the acidity, viscosity, and number of Streptococcus thermophilus increased with increasing concentration of additive added in the experimental tests. Indeed, it seems clear that the quality of fermented milk and storability is more improved than the incorporation rate is high. The products showed a better test and a firmer texture limiting the whey exudation.

Keywords: fermented milk, pectin, gelatin, whey proteins, functional properties, quality, conservation, valorization

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88 Production of Novel Antibiotics of Tylosin by Importing eryK and eryG Genes in Streptomyces fradiae

Authors: Neda Gegar Goshe, M. Moradi, Hossein Rassi


The antibacterial properties of macrolide antibiotics (such as erythromycin and tylosin) depend ultimately on the glycosylation of otherwise inactive polyketide lactones. Among the sugars commonly found in such macrolides are various 6-deoxyhexoses including the 3-dimethylamino sugars mycaminose and desosamine (4-deoxymycaminose). Some macrolides (such as tylosin) possess multiple sugar moieties, whereas others (such as erythromycin) have two sugar substituents. Streptomyces fradiae is an ideal host for development of generic polyketide-overproducing strains because it contains three of the most common precursors-malonyl-CoA, methylmalonyl-CoA and ethylmalonyl-CoA-used by modular PKS, and is a host that is amenable to genetic manipulation. As patterns of glycosylation markedly influence a macrolide's drug activity, there is considerable interest in the possibility of using combinatorial biosynthesis to generate new pairings of polyketide lactones with sugars, especially 6-deoxyhexoses. Here, we report a successful attempt to alter the aminodeoxyhexose-biosynthetic capacity of Streptomyces fradiae (a producer of tylosin) by importing genes from the erythromycin producer Saccharopolyspora erythraea. The bio transformation of erythromycin-D into the desired major component erythromycin-A involves two final enzymatic reactions, EryK-catalyzed hydroxylation at the C-12 position of the aglycone and EryG-catalyzed O methylation at the C-3 position of macrose. This engineered S. fradiae produced substantial amounts of two potentially useful macrolides that had not previously been obtained by fermentation.

Keywords: tylosin, eryK and eryG genes, streptomyces fradiae

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87 Effects of Multilayer Coating of Chitosan and Polystyrene Sulfonate on Quality of ‘Nam Dok Mai No.4’ Mango

Authors: N. Hadthamard, P. Chaumpluk, M. Buanong, P. Boonyaritthongchai, C. Wongs-Aree


Ripe ‘Nam Dok Mai’ mango (Mangifera indica L.) is an important exported fruit of Thailand, but rapidly declined in the quality attributes mainly by infection of anthracnose and stem end rot diseases. Multilayer coating is considered as a developed technique to maintain the postharvest quality of mangoes. The utilization of alternated coating by matching oppositely electrostatic charges between 0.1% chitosan and 0.1% polystyrene sulfonate (PSS) was studied. A number of the coating layers (layer by layer) were applied on mature green ‘Nam Dok Mai No.4’ mangoes prior to storage at 25 oC, 65-70% relative humidity (RH). There were significant differences in some quality attributes of mangoes coated by 3½ layers, 4½ layers and 5½ layers. In comparison to coated mangoes, uncoated fruits were higher in weight loss, total soluble solids, respiration rate, ethylene production and disease incidence except the titratable acidity. Coating fruit at 3½ layers exhibited the ripening delay and reducing disease infection without off flavour. On the other hand, fruit coated with 5½ layers comprised the lowest acceptable score, caused by exhibiting disorders from fermentation at the end of storage. As a result, multilayer coating between chitosan and PSS could effectively maintain the postharvest quality of mango, but number of coating layers should be thoroughly considered.

Keywords: multilayer, chitosan, polystyrene sulfonate, Nam Dok Mai No.4

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86 Encapsulated Rennin Enzyme in Nano and Micro Tubular Cellulose/Starch Gel Composite for Milk Coagulation

Authors: Eleftheria Barouni, Theano Petsi, Argyro Bekatorou, Dionysos Kolliopoulos, Dimitrios Vasileiou, Panayiotis Panas, Maria Kanellaki, Athanasios A. Koutinas


The aim of the present work was the production and use of a composite filter (TC/starch), containing rennin enzyme, in continuous system and in successive fermentation batches (SFB) for milk coagulation in order to compare the operational stability of both systems and cheese production cost. Tubular cellulose (TC) was produced after removal of lignin from lignocellulosic biomass using several procedures, e.g. alkaline treatment [1] and starch gel was added for the reduction of TC tubes dimensions to micro- and nano- range[2]. Four immobilized biocatalysts were prepared using different ways of the enzyme entrapment. 1) TC/ rennin (rennin entrapped in the tubes of TC), 2) TC/SG-rennin (rennin entrapped in the tubes of the composite), 3) TC-SG/rennin (rennin entrapped into the layer of starch gel) and 4) TC/rennin- SG/rennin (rennin is entrapped both in the tubes of the TC and into the layer of starch gel). Firstly these immobilized biocatalysts were examined in ten SFB regarding the coagulation time and their activity All the above immobilized biocatalysts remained active and the coagulation time was ranged from 90 to 480, 120-480, 330-510, and 270-540 min for (1), (2), (3), and (4) respectively. The quality of the cheese was examined through the determination of volatile compounds by SPME GC/MS analysis. These results encouraged us to study a continuous coagulation system of milk. Even though the (1) immobilized biocatalyst gave lower coagulation time, we used the (2) immobilized biocatalyst in the continuous system. The results were promising.

Keywords: tubular cellulose, starch gel, composite biocatalyst, Rennin, milk coagulation

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85 Impact of a Locally-Prepared Fermented Alcoholic Beverage from Jaggery on the Gut Bacterial Profile of the Tea-Tribal Populations of Assam, India

Authors: Rupamoni Thakur, Madhusmita Dehingia, Narayan C. Talukdar, Mojibur R. Khan


The human gut is an extremely active fermentation site and is inhabited by diverse bacterial species. Consumption of alcoholic beverages has been shown to substantially modulate the human gut bacterial profile (GBP) of an individual. Assam, a major north-eastern state of India, is home to a number of tribal populations of which the tea-tribes form a major community. These tea-tribal communities are known to prepare and consume a locally-prepared alcoholic beverage from fermented jaggery, whose chemical composition is unknown. In this study, we demonstrate the effect of daily intake of the locally-prepared alcoholic beverage on the GBP of the tea-tribal communities and correlate it with the changes in the biochemical biomarkers of the population. The fecal bacterial diversity of 40 drinkers and 35 non-drinking healthy individuals were analyzed by polymerase chain reaction (PCR)–denaturing gradient gel electrophoresis (DGGE). The results suggested that the GBP was significantly modulated in the fermented-beverage consuming subjects. Significant difference was also observed in the serum biochemical parameters such as triglyceride, total cholesterol and the liver marker enzymes (ASAT/ALAT and GGT). Further studies to identify the GBP of drinkers vs non-drinkers through Next-generation Sequencing (NGS) analysis and to correlate the changes with the biochemical biomarkers of the population is underway.

Keywords: alcoholic beverage, gut bacterial profile, PCR-DGGE analysis, tea-tribes of India

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84 Improvement Anaerobic Digestion Performance of Sewage Sludge by Co-Digestion with Cattle Manure

Authors: Raouf Hassan


Biogas energy production from sewage sludge is an economically feasible and eco-friendly in nature. Sewage sludge is considered nutrient-rich substrates, but had lower values of carbone which consider an energy source for anaerobic bacteria. The lack or lower values of carbone-to-nitrogen ratio (C/N) reduced biogas yield and fermentation rate. Anaerobic co-digestion of sewage sludge offers several benefits over mono-digestion such as optimize nutrient balance, increased cost-efficiency and increased degradation rate. The high produced amounts of animal manures, which reach up to 90% of the total collected organic wastes, are recommended for the co-digestion with sewage sludge, especially with the limitations of industrial substrates. Moreover, cattle manures had high methane production potential (500 m3/t vsadded). When mixed with sewage sludge the potential methane production increased with increasing cattle manure content. In this paper, the effect of cattle manure (CM) addition as co-substrates on the sewage sludge (SS) anaerobic digestion performance was investigated under mesophilic conditions (35°C) using anaerobic batch reactors. The batch reactors were operated with a working volume 0.8 liter, and a hydraulic retention time of 30 days. The research work focus on studying two main parameters; the biogas yield (expressed as VSS) and pH values inside the reactors.

Keywords: anaerobic digestion, sewage sludge, cattle manure, mesophilic, biogas yield, pH

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83 Anti Staphylococcus aureus and Methicillin Resistant Staphylococcus aureus Action of Thermophilic Fungi Acrophialophora levis IBSD19 and Determination of Its Mode of Action Using Electron Microscopy

Authors: Shivankar Agrawal, Indira Sarangthem


Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA) remains one of the major causes of healthcare-associated and community-onset infections worldwide. Hence the search for non-toxic natural compounds having antibacterial activity has intensified for future drug development. The exploration of less studied niches of Earth can highly increase the possibility to discover novel bioactive compounds. Therefore, in this study, the cultivable fraction of fungi from the sediments of natural hot springs has been studied to mine potential fungal candidates with antibacterial activity against the human pathogen Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. We isolated diverse strains of thermophilic fungi from a collection of samples from sediment. Following a standard method, we isolated a promising thermophilic fungus strain IBSD19, identified as Acrophialophora levis, possessing the potential to produce an anti-Staphylococcus aureus agent. The growth conditions were optimized and scaled to fermentation, and its produced extract was subjected to chemical extraction. The ethyl acetate fraction was found to display significant activity against Staphylococcus aureus and MRSA with a minimum inhibitory concentration (MIC) of 0.5 mg/ml and 4 mg/ml, respectively. The cell membrane integrity assay and SEM suggested that the fungal metabolites cause bacteria clustering and further lysis of the cell.

Keywords: antibacterial activity, antioxidant, fungi, Staphylococcus aureus, MRSA, thermophiles

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