Search results for: alcoholic fermentation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 540

Search results for: alcoholic fermentation

90 Rheological Properties of Dough and Sensory Quality of Crackers with Dietary Fibers

Authors: Ljubica Dokić, Ivana Nikolić, Dragana Šoronja–Simović, Zita Šereš, Biljana Pajin, Nils Juul, Nikola Maravić

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The possibility of application the dietary fibers in production of crackers was observed in this work, as well as their influence on rheological and textural properties on the dough for crackers and influence on sensory properties of obtained crackers. Three different dietary fibers, oat, potato and pea fibers, replaced 10% of wheat flour. Long fermentation process and baking test method were used for crackers production. The changes of dough for crackers were observed by rheological methods of determination the viscoelastic dough properties and by textural measurements. Sensory quality of obtained crackers was described using quantity descriptive method (QDA) by trained members of descriptive panel. Additional analysis of crackers surface was performed by videometer. Based on rheological determination, viscoelastic properties of dough for crackers were reduced by application of dietary fibers. Manipulation of dough with 10% of potato fiber was disabled, thus the recipe modification included increase in water content at 35%. Dough compliance to constant stress for samples with dietary fibers decreased, due to more rigid and stiffer dough consistency compared to control sample. Also, hardness of dough for these samples increased and dough extensibility decreased. Sensory properties of final products, crackers, were reduced compared to control sample. Application of dietary fibers affected mostly hardness, structure and crispness of the crackers. Observed crackers were low marked for flavor and taste, due to influence of fibers specific aroma. The sample with 10% of potato fibers and increased water content was the most adaptable to applied stresses and to production process. Also this sample was close to control sample without dietary fibers by evaluation of sensory properties and by results of videometer method.

Keywords: crackers, dietary fibers, rheology, sensory properties

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89 Egyptian Soil Isolate Shows Promise as a Source of a New Broad-spectrum Antimicrobial Agent Against Multidrug-resistant Pathogens

Authors: Norhan H. Mahdally, Bathini Thissera Riham A. ElShiekh, Noha M. Elhosseiny, Mona T. Kashef, Ali M. El Halawany, Mostafa E. Rateb, Ahmed S. Attia

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Multidrug-resistant (MDR) pathogens pose a global threat to healthcare settings. The exhaustion of the current antibiotic arsenal and the scarcity of new antimicrobials in the pipeline aggravate this threat and necessitate a prompt and effective response. This study focused on two major pathogens that can cause serious infections: carbapenem-resistant Acinetobacter baumannii (CRAB) and methicillin-resistant Staphylococcus aureus (MRSA). Multiple soil isolates were collected from several locations throughout Egypt and screened for their conventional and non-conventional antimicrobial activities against MDR pathogens. One isolate exhibited potent antimicrobial activity and was subjected to multiple rounds of fractionation. After fermentation and bio-guided fractionation, we identified pure microbial secondary metabolites with two scaffolds that exhibited promising effects against CRAB and MRSA. Scaling up and chemical synthesis of derivatives of the identified metabolite resulted in obtaining a more potent derivative, which we designated as 2HP. Cytotoxicity studies indicated that 2HP is well-tolerated by human cells. Ongoing work is focusing on formulating the new compound into a nano-formulation to enhance its delivery. Also, to have a better idea about how this compound works, a proteomic approach is currently underway. Our findings suggest that 2HP is a potential new broad-spectrum antimicrobial agent. Further studies are needed to confirm these findings and to develop 2HP into a safe and effective treatment for MDR infections.

Keywords: broad-spectrum antimicrobials, carbapenem-resistant acinetobacter baumannii, drug discovery, methicillin-resistant staphylococcus aureus, multidrug-resistant, natural products

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88 Repeated Batch Cultivation: A Novel Empty and Fill Strategy for the Enhanced Production of a Biodegradable Polymer, Polyhydroxy Alkanoate by Alcaligenes latus

Authors: Geeta Gahlawat, Ashok Kumar Srivastava

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In the present study, a simple drain and fill protocol strategy of repeated batch was adopted for enhancement in polyhydroxyalkanoates (PHAs) production using alcaligenes latus DSM 1124. Repeated batch strategy helped in increasing the longevity of otherwise decaying culture in the bioreactor by supplementing fresh substrates during each cycle of repeated-batch. The main advantages of repeated batch are its ease of operation, enhancement of culture stability towards contamination, minimization of pre-culture effects and maintenance of organism at high growth rates. The cultivation of A. latus was carried out in 7 L bioreactor containing 4 L optimized nutrient medium and a comparison with the batch mode fermentation was done to evaluate the performance of repeated batch in terms of PHAs accumulation and productivity. The statistically optimized medium recipe consisted of: 25 g/L Sucrose, 2.8 g/L (NH4)2SO4, 3.25 g/L KH2PO4, 3.25 g/L Na2HPO4, 0.2 g/L MgSO4, 1.5 mL/L trace element solution. In this strategy, 20% (v/v) of the culture broth was removed from the reactor and supplemented with an equal volume of fresh medium when sucrose concentration inside the reactor decreased below 8 g/L. The fermenter was operated for three repeated batch cycles and fresh nutrient feeding was done at 27 h, 48 h, and 60 h. Repeated batch operation resulted in a total biomass of 27.89 g/L and PHAs concentration 20.55 g/L at the end of 69 h which was a marked improvement as compared to batch cultivation (8.71 g/L biomass and 6.24 g/L PHAs). This strategy demonstrated 3.3 fold and 1.8 fold increase in PHAs concentration and volumetric productivity, respectively as compared to batch cultivation. Repeated batch cultivation strategy had also the benefit of avoiding non-productive time period required for cleaning, refilling and sterilization of bioreactor, thereby increasing the overall volumetric productivity and making the entire process cost-effective too.

Keywords: alcaligenes, biodegradation, polyhydroxyalkanoates, repeated batch

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87 Characterization of Pectinase from Local Microorganisms to Support Industry Based Green Chemistry

Authors: Sasangka Prasetyawan, Anna Roosdiana, Diah Mardiana, Suratmo

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Pectinase are enzymes that hydrolyze pectin compounds. The use of this enzyme is primarily to reduce the viscosity of the beverage thus simplifying the purification process. Pectinase activity influenced by microbial sources . Exploration of two types of microbes that Aspergillus spp. and Bacillus spp. pectinase give different performance, but the use of local strain is still not widely studied. The aim of this research is exploration of pectinase from A. niger and B. firmus include production conditions and characterization. Bacillus firmus incubated and shaken at a speed of 200 rpm at pH variation (5, 6, 7, 8, 9, 10), temperature (30, 35, 40, 45, 50) °C and incubation time (6, 12, 18, 24, 30, 36 ) hours. Media was centrifuged at 3000 rpm, pectinase enzyme activity determined. Enzyme production by A. niger determined to variations in temperature and pH were similar to B. firmus, but the variation of the incubation time was 24, 48, 72, 96, 120 hours. Pectinase crude extract was further purified by precipitation using ammonium sulfate saturation in fraction 0-20 %, 20-40 %, 40-60 %, 60-80 %, then dialyzed. Determination of optimum conditions pectinase activity performed by measuring the variation of enzyme activity on pH (4, 6, 7, 8, 10), temperature (30, 35, 40, 45, 50) °C, and the incubation time (10, 20, 30, 40, 50) minutes . Determination of kinetic parameters of pectinase enzyme reaction carried out by measuring the rate of enzyme reactions at the optimum conditions, but the variation of the concentration of substrate (pectin 0.1 % , 0.2 % , 0.3 % , 0.4 % , 0.5 % ). The results showed that the optimum conditions of production of pectinase from B. firmus achieved at pH 7-8.0, 40-50 ⁰C temperature and fermentation time 18 hours. Purification of pectinase showed the highest purity in the 40-80 % ammonium sulfate fraction. Character pectinase obtained : the optimum working conditions of A. niger pectinase at pH 5 , while pectinase from B. firmus at pH 7, temperature and optimum incubation time showed the same value, namely the temperature of 50 ⁰C and incubation time of 30 minutes. The presence of metal ions can affect the activity of pectinase , the concentration of Zn 2 + , Pb 2 + , Ca 2 + and K + and 2 mM Mg 2 + above 6 mM inhibit the activity of pectinase .

Keywords: pectinase, Bacillus firmus, Aspergillus niger, green chemistry

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86 Impact of Helicobacter pylori Infection on Colorectal Adenoma-Colorectal Carcinoma Sequence

Authors: Jannis Kountouras, Nikolaos Kapetanakis, Stergios A. Polyzos, Apostolis Papaeftymiou, Panagiotis Katsinelos, Ioannis Venizelos, Christina Nikolaidou, Christos Zavos, Iordanis Romiopoulos, Elena Tsiaousi, Evangelos Kazakos, Michael Doulberis

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Background & Aims: Helicobacter pylori infection (Hp-I) has been recognized as a substantial risk agent involved in gastrointestinal (GI) tract oncogenesis by stimulating cancer stem cells (CSCs), oncogenes, immune surveillance processes, and triggering GI microbiota dysbiosis. We aimed to investigate the possible involvement of active Hp-I in the sequence: chronic inflammation–adenoma–colorectal cancer (CRC) development. Methods: Four pillars were investigated: (i) endoscopic and conventional histological examinations of patients with CRC, colorectal adenomas (CRA) versus controls to detect the presence of active Hp-I; (ii) immunohistochemical determination of the presence of Hp; expression of CD44, an indicator of CSCs and/or bone marrow-derived stem cells (BMDSCs); expressions of oncogene Ki67 and anti-apoptotic Bcl-2 protein; (iii) expression of CD45, indicator of immune surveillance locally (assessing mainly T and B lymphocytes locally); and (iv) correlation of the studied parameters with the presence or absence of Hp-I. Results: Among 50 patients with CRC, 25 with CRA, and 10 controls, a significantly higher presence of Hp-I in the CRA (68%) and CRC group (84%) were found compared with controls (30%). The presence of Hp-I with accompanying immunohistochemical expression of CD44 in biopsy specimens was revealed in a high proportion of patients with CRA associated with moderate/severe dysplasia (88%) and CRC patients with moderate/severe degree of malignancy (91%). Comparable results were also obtained for Ki67, Bcl-2, and CD45 immunohistochemical expressions. Concluding Remarks: Hp-I seems to be involved in the sequence: CRA – dysplasia – CRC, similarly to the upper GI tract oncogenesis, by several pathways such as the following: Beyond Hp-I associated insulin resistance, the major underlying mechanism responsible for the metabolic syndrome (MetS) that increase the risk of colorectal neoplasms, as implied by other Hp-I related MetS pathologies, such as non-alcoholic fatty liver disease and upper GI cancer, the disturbance of the normal GI microbiota (i.e., dysbiosis) and the formation of an irritative biofilm could contribute to a perpetual inflammatory upper GIT and colon mucosal damage, stimulating CSCs or recruiting BMDSCs and affecting oncogenes and immune surveillance processes. Further large-scale relative studies with a pathophysiological perspective are necessary to demonstrate in-depth this relationship.

Keywords: Helicobacter pylori, colorectal cancer, colorectal adenomas, gastrointestinal oncogenesis

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85 Bacteriological Quality of Commercially Prepared Fermented Ogi (AKAMU) Sold in Some Parts of South Eastern Nigeria

Authors: Alloysius C. Ogodo, Ositadinma C. Ugbogu, Uzochukwu G. Ekeleme

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Food poisoning and infection by bacteria are of public health significance to both developing and developed countries. Samples of ogi (akamu) prepared from white and yellow variety of maize sold in Uturu and Okigwe were analyzed together with the laboratory prepared ogi for microbial quality using the standard microbiological methods. The analyses showed that both white and yellow variety had total bacterial counts (cfu/g) of 4.0 ×107 and 3.9 x 107 for the laboratory prepared ogi while the commercial ogi had 5.2 x 107 and 4.9 x107, 4.9 x107 and 4.5 x107, 5.4 x107 and 5.0 x107 for Eke-Okigwe, Up-gate and Nkwo-Achara market respectively. The Staphylococcal counts ranged from 2.0 x 102 to 5.0 x102 and 1.0 x 102 to 4.0 x102 for the white and yellow variety from the different markets while Staphylococcal growth was not recorded on the laboratory prepared ogi. The laboratory prepared ogi had no Coliform growth while the commercially prepared ogi had counts of 0.5 x103 to 1.6 x 103 for white variety and 0.3 x 103 to 1.1 x103 for yellow variety respectively. The Lactic acid bacterial count of 3.5x106 and 3.0x106 was recorded for the laboratory ogi while the commercially prepared ogi ranged from 3.2x106 to 4.2x106 (white variety) and 3.0 x106 to 3.9 x106 (yellow). The presence of bacteria isolates from the commercial and laboratory fermented ogi showed that Lactobacillus sp, Leuconostoc sp and Citrobacter sp were present in all the samples, Micrococcus sp and Klebsiella sp were isolated from Eke-Okigwe and ABSU-up-gate markets varieties respectively, E. coli and Staphylococcus sp were present in Eke-Okigwe and Nkwo-Achara markets while Salmonella sp were isolated from the three markets. Hence, there are chances of contracting food borne diseases from commercially prepared ogi. Therefore, there is the need for sanitary measures in the production of fermented cereals so as to minimize the rate of food borne pathogens during processing and storage.

Keywords: ogi, fermentation, bacterial quality, lactic acid bacteria, maize

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84 Nitrogen Fixation in Hare Gastrointestinal Tract

Authors: Tatiana A. Kuznetsova, Maxim V. Vechersky, Natalia V. Kostina, Marat M. Umarov, Elena I. Naumova

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One of the main problems of nutrition of phytophagous animals is the insufficiency of protein in their forage. Usually, symbiotic microorganisms highly contribute both to carbohydrates and nitrogen compounds of the food. But it is not easy to utilize microbial biomass in the large intestine and caecum for the animals with hindgut fermentation. So that, some animals, as well hares, developed special mechanism of contribution of such biomass - obligate autocoprophagy, or reingestion. Hares have two types of feces - the hard and the soft. Hard feces are excreted at night, while hares are vigilance ("foraging period"), and the soft ones (caecotrophs) are produced and reingested in the day-time during hares "resting-period". We examine the role of microbial digestion in providing nitrogen nutrition of hare (Lepus europaeus). We determine the ability of nitrogen fixation in fornix and stomach body, small intestine, caecum and colon of hares' gastro-intestinal tract in two main period of hares activity - "resting-period" (day time) and "foraging period" (late-evening and very-early-morning). We use gas chromatography to measure levels of nitrogen fixing activity (acetylene reduction). Nitrogen fixing activity was detected in the contents of all analyzed parts of the gastrointestinal tract. Maximum values were recorded in the large intestine. Also daily dynamics of the process was detected. Thus, during hare “resting-period” (caecotrophs formation) N2-fixing activity was significantly higher than during “foraging period”, reaching 0,3 nmol C2H4/g*h. N2-fixing activity in the gastrointestinal tract can allocate to significant contribution of nitrogen fixers to microbial digestion in hare and confirms the importance of coprophagy as a nitrogen source in lagomorphs.

Keywords: coprophagy, gastrointestinal tract, lagomorphs, nitrogen fixation

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83 Antimicrobial Activity of Endophytes on some Selected Clinical Isolates (Escherichia coli, Staphylococcus aureus, Salmonella Typhi, Bacillus subtilis, Klebsiella pneumoniae, Aspergillus fumigatus, Pseudomomonas aeruginosa and Penicillium chryysogenum)

Authors: Dawang D. N., Dasat G. S., Nden D.

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Endophyte means “in the plant” are referred to all microorganisms that live in the internal tissues of stems, petioles, roots and leaves of plants causing no apparent symptoms of disease. Secondary metabolites from fungal endophytes have an enormous potential applications as antioxidant, antimicrobial, anticancer and antidiabeties. Thus, this study aimed to determine the antimicrobial activity of these metabolites against some clinical isolates. The fungi were subjected to fermentation medium and the metabolites were extracted using ethyl acetate. The fungal extracts showed both antibacterial and antifungal activities with maximum zone of inhibition diameter of 10.5mm against Aspergillus fumigatus. Staphylococcus aureus was inhibited by all the five crude extracts with inhibition zone diameter of 4mm. Endophytic fungal crude extract2 (EDF2) exhibited antimicrobial effect against all the test organisms used, EDF4 was active against all test organisms except on Penicillium chrysogenum and Klebsiella pneumoniae. Antibacterial standard of ciprofloxacin which is 15mm is comparable to the effect of endophytic extract of EDF1 and EDF2. Klebsiella pneumoniae was resistant to EDF4 and EDF5. EDF3 showed a wide range of antimicrobial activity against all the test organisms used. The highest inhibition zone diameter of 10.50mm recorded against Aspergillus fumigatus is comparable to antifungal standard of fluconazole (15.5mm). The result of this study suggests that endophytic fungi associated with the roots of Irish potato could be a promising source of novel bioactive compounds of pharmaceutical and industrial importance.

Keywords: endophyte, fungal extract, antimicrobial, potato

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82 A Combinatorial Approach of Treatment for Landfill Leachate

Authors: Anusha Atmakuri, R. D. Tyagi, Patrick Drogui

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Landfilling is the most familiar and easy way to dispose solid waste. Landfill is generally received via wastes from municipal near to a landfill. The waste collected is from commercial, industrial, and residential areas and many more. Landfill leachate (LFL) is formed when rainwater passes through the waste placed in landfills and consists of several dissolved organic materials, for instance, aquatic humic substances (AHS), volatile fatty acids (VFAs), heavy metals, inorganic macro components, and xenobiotic organic matters, highly toxic to the environment. These components of LFL put a load on it, hence it necessitates the treatment of LFL prior to its discharge into the environment. Various methods have been used to treat LFL over the years, such as physical, chemical, biological, physicochemical, electrical, and advanced oxidation methods. This study focuses on the combination of biological and electrochemical methods- extracellular polymeric substances and electrocoagulation(EC). The coupling of electro-coagulation process with extracellular polymeric substances (EPS) (as flocculant) as pre and\or post treatment strategy provides efficient and economical process for the decontamination of landfill leachate contaminated with suspended matter, metals (e.g., Fe, Mn) and ammonical nitrogen. Electro-coagulation and EPS mediated coagulation approach could be an economically viable for the treatment of landfill leachate, along with possessing several other advantages over several other methods. This study utilised waste substrates such as activated sludge, crude glycerol and waste cooking oil for the production of EPS using fermentation technology. A comparison of different scenarios for the treatment of landfill leachate is presented- such as using EPS alone as bioflocculant, EPS and EC with EPS being the 1st stage, and EPS and EC with EC being the 1st stage. The work establishes the use of crude EPS as a bioflocculant for the treatment of landfill leachate and wastewater from a site near a landfill, along with EC being successful in removal of some major pollutants such as COD, turbidity, total suspended solids. A combination of these two methods is to be explored more for the complete removal of all pollutants from landfill leachate.

Keywords: landfill leachate, extracellular polymeric substances, electrocoagulation, bioflocculant.

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81 Development of Broad Spectrum Nitrilase Biocatalysts and Bioprocesses for Nitrile Biotransformation

Authors: Avinash Vellore Sunder, Shikha Shah, Pramod P. Wangikar

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The enzymatic conversion of nitriles to carboxylic acids by nitrilases has gained significance in the green synthesis of several pharmaceutical precursors and fine chemicals. While nitrilases have been characterized from different sources, the industrial application requires the identification of nitrilases that possess higher substrate tolerance, wider specificity and better thermostability, along with the development of an efficient bioprocess for producing large amounts of nitrilase. To produce large amounts of nitrilase, we developed a fed-batch fermentation process on defined media for the high cell density cultivation of E. coli cells expressing the well-studied nitrilase from Alcaligenes fecalis. A DO-stat feeding approach was employed combined with an optimized post-induction strategy to achieve nitrilase titer of 2.5*105 U/l and 78 g/l dry cell weight. We also identified 16 novel nitrilase sequences from genome mining and analysis of substrate binding residues. The nitrilases were expressed in E. coli and their biocatalytic potential was evaluated on a panel of 22 industrially relevant nitrile substrates using high-throughput screening and HPLC analysis. Nine nitrilases were identified to exhibit high activity on structurally diverse nitriles including aliphatic and aromatic dinitriles, heterocyclic, -hydroxy and -keto nitriles. With fed-batch biotransformation, whole-cell Zobelia galactanivorans nitrilase achieved yields of 2.4 M nicotinic acid and 1.8 M isonicotinic acid from 3-cyanopyridine and 4-cyanopyridine respectively within 5 h, while Cupravidus necator nitrilase enantioselectively converted 740 mM mandelonitrile to (R)–mandelic acid. The nitrilase from Achromobacter insolitus could hydrolyze 542 mM iminodiacetonitrile in 1 h. The availability of highly active nitrilases along with bioprocesses for enzyme production expands the toolbox for industrial biocatalysis.

Keywords: biocatalysis, isonicotinic acid, iminodiacetic acid, mandelic acid, nitrilase

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80 Fractionation of Biosynthetic Mixture of Gentamicins by Reactive Extraction

Authors: L. Kloetzer, M. Poştaru, A. I. Galaction, D. Caşcaval

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Gentamicin is an aminoglycoside antibiotic industrially obtained by biosynthesis of Micromonospora purpurea or echinospora, the product being a complex mixture of components with very similar structures. Among them, three exhibit the most important biological activity: gentamicins C1, C1a, C2, and C2a. The separation of gentamicin from the fermentation broths at industrial scale is rather difficult and it does not allow the fractionation of the complex mixture of gentamicins in order to increase the therapeutic activity of the product. The aim of our experiments is to analyze the possibility to selectively separate the less active gentamicin, namely gentamicin C1, from the biosynthetic mixture by reactive extraction with di-(2-ethylhexyl) phosphoric acid (D2EHPA) dissolved in dichloromethane, followed selective re-extraction of the most active gentamicins C1a, C2, and C2a. The experiments on the reactive extraction of gentamicins indicated the possibility to separate selectively the gentamicin C1 from the mixture obtained by biosynthesis. The extraction selectivity is positively influenced by increasing the pH-value of an aqueous solution and by using a D2EHPA concentration in organic phase closer to the value needed for an equimolecular ratio between the extractant and this gentamicin. For quantifying the selectivity of separation, the selectivity factor, calculated as the ratio between the degree of reactive extraction of gentamicin C1 and the overall extraction degree of gentamicins were used. The possibility to remove the gentamicin C1 at an extractant concentration of 10 g l-1 and pH = 8 is presented. In these conditions, it was obtained the maximum value of the selectivity factor of 2.14, which corresponds to the modification of the gentamicin C1 concentration from 31.92% in the biosynthetic mixture to 72% in the extract. The re-extraction of gentamicins C1, C1a, C2, and C2a with sulfuric acid from the extract previously obtained by reactive extraction (mixture A – extract obtained by non-selective reactive extraction; mixture B – extract obtained by selective reactive extraction) allows for separating selectively the most active gentamicins C1a, C2, and C2a. For recovering only the active gentamicins C1a, C2, and C2a, the re-extraction must be carried out at very low acid concentrations, far below those corresponding to the stoichiometry of its chemical reactions with these gentamicins. Therefore, the mixture resulted by re-extraction contained 92.6% gentamicins C1a, C2, and C2a. By bringing together the aqueous solutions obtained by reactive extraction and re-extraction, the overall content of the active gentamicins in the final product becomes 89%, their loss reaching 0.3% related to the initial biosynthetic product.

Keywords: di-(2-ethylhexyl) phosphoric acid, gentamicin, reactive extraction, selectivity factor

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79 The Changes of Chemical Composition of Rice Straw Treated by a Biodecomposer Developed from Rumen Bacterial of Buffalo

Authors: A. Natsir, M. Nadir, S. Syahrir, A. Mujnisa

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In tropical countries such as in Indonesia, rice straw plays an important role in fulfilling the needs of feed for ruminant, especially during the dry season in which the availability of forage is very limited. However, the main problem of using rice straw as a feedstuff is low digestibility due to the existence of the links between lignin and cellulose or hemicellulose, and imbalance of its minerals content. One alternative to solve this problem is by application of biodecomposer (BS) derived from rumen bacterial of the ruminant. This study was designed to assess the effects of BS application on the changes of the chemical composition of rice straw. Four adults local buffalo raised under typical feeding conditions were used as a source of inoculum for BS development. The animal was fed for a month with a diet consisted of rice straw and elephant grass before taking rumen fluid samples. Samples of rumen fluid were inoculated in the carboxymethyl cellulose (CMC) media under anaerobic condition for 48 hours at 37°C. The mixture of CMC media and microbes are ready to be used as a biodecomposer following incubation of the mixture under anaerobic condition for 7 days at 45°C. The effectiveness of BS then assessed by applying the BS on the straw according to completely randomized design consisted of four treatments and three replication. One hundred g of ground coarse rice straw was used as the substrate. The BS was applied to the rice straw substrate with the following composition: Rice straw without BS (P0), rice straw + 5% BS (P1), rice straw +10% BS (P2), and rice straw + 15% BS. The mixture of rice straw and BS then fermented under anaerobic for four weeks. Following the fermentation, the chemical composition of rice straw was evaluated. The results indicated that the crude protein content of rice straw significantly increased (P < 0.05) as the level of BS increased. On the other hand, the concentration of crude fiber of the rice straw was significantly decreased (P < 0.05) as the level of BS increased. Other nutrients such as minerals did not change (P > 0.05) due to the treatments. In conclusion, application of BS developed from rumen bacterial of buffalo has a promising prospect to be used as a biological agent to improve the quality of rice straw as feeding for ruminant.

Keywords: biodecomposer, local buffalo, rumen microbial, chemical composition

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78 Investigation of Enterotoxigenic Staphylococcus aureus in Kitchen of Catering

Authors: Çiğdem Sezer, Aksem Aksoy, Leyla Vatansever

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This study has been done for the purpose of evaluation of public health and identifying of enterotoxigenic Staphyloccocus aureus in kitchen of catering. In the kitchen of catering, samples have been taken by swabs from surface of equipments which are in the salad section, meat section and bakery section. Samples have been investigated with classical cultural methods in terms of Staphyloccocus aureus. Therefore, as a 10x10 cm area was identified (salad, cutting and chopping surfaces, knives, meat grinder, meat chopping surface) samples have been taken with sterile swabs with helping FTS from this area. In total, 50 samples were obtained. In aseptic conditions, Baird-Parker agar (with egg yolk tellurite) surface was seeded with swabs. After 24-48 hours of incubation at 37°C, the black colonies with 1-1.5 mm diameter and which are surrounded by a zone indicating lecithinase activity were identified as S. aureus after applying Gram staining, catalase, coagulase, glucose and mannitol fermentation and termonuclease tests. Genotypic characterization (Staphylococcus genus and S.aureus species spesific) of isolates was performed by PCR. The ELISA test was applied to the isolates for the identification of staphylococcal enterotoxins (SET) A, B, C, D, E in bacterial cultures. Measurements were taken at 450 nm in an ELISA reader using an Ridascreen-Total set ELISA test kit (r-biopharm R4105-Enterotoxin A, B, C, D, E). The results were calculated according to the manufacturer’s instructions. A total of 50 samples of 97 S. aureus was isolated. This number has been identified as 60 with PCR analysis. According to ELISA test, only 1 of 60 isolates were found to be enterotoxigenic. Enterotoxigenic strains were identified from the surface of salad chopping and cutting. In the kitchen of catering, S. aureus identification indicates a significant source of contamination. Especially, in raw consumed salad preparation phase of contamination is very important. This food can be a potential source of food-borne poisoning their terms, and they pose a significant risk to consumers have been identified.

Keywords: Staphylococcus aureus, enterotoxin, catering, kitchen, health

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77 Non-Candida Albicans Candida: Virulence Factors and Species Identification in India

Authors: Satender Saraswat, Dharmendra Prasad Singh, Rajesh Kumar Verma, Swati Sarswat

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Background and Purpose: The predominant cause of candidiasis was Candida albicans which has shifted towards non-Candida albicans Candida (NCAC) (Candida species other than the C. albicans). NCAC, earlier considered non-pathogenic or minimally virulent, are now considered a primary cause of morbidity and mortality in immunocompromised. With the NCAC spp. gaining weightage in the clinical cases, this study was conducted to determine the prevalence of NCAC spp. in different clinical specimens and to assess a few of their virulence factors. Material and Methods: Routine samples for bacterial culture and sensitivity, showing colony characteristics like Candida on Blood Agar and microscopic features resembling Candida spp. were processed further. Candida isolates were tested for chlamydospore formation, biochemical tests including sugar fermentation and sugar assimilation tests, and growth at 42oC, colony colour on HiCrome™ Candida Differential Agar, HiCandida Identification Kit and VITEK-2 Compact. Virulence factors like adherence to buccal epithelial cells (ABEC), biofilm formation, hemolytic activity, and production of coagulase enzyme were also tested. Results: Mean age of the patients was 38.46 with a male-female ratio of 1.36:1. 137 Candida isolates were recovered. 45.3% isolates were isolated from urine, 19.7% from vaginal swabs and 13.9% from oropharyngeal swabs. 55 (40.1%) isolates of C. albicans and 82 (59.9%) of NCAC spp. were identified, with C. tropicalis (23.4%) in NCAC. C. albicans (3; 50%) was the commonest species in cases of candidemia. Haemolysin production (85.5%) and ABEC (78.2%) were the major virulence factors in C. albicans. C. tropicalis (59.4%) and C. dubliniensis (50%) showed maximum ABEC. Biofilm forming capacity was higher in C. tropicalis (78.1%) than C. albicans (67%). Conclusion: This study suggests varied prevalence and virulence based on geographical locations, even within a subcontinent. It clearly demarcates the emergence of NCAC and their predominance in different body fluids. Identification of Candida to species level should become a routine in all the laboratories.

Keywords: ABEC, NCAC, non-Candida albicans Candida, Vitek-2TM compact

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76 Laboratory Scale Production of Bio-Based Chemicals from Industrial Waste Feedstock in South Africa

Authors: P. Mandree, S. O. Ramchuran, F. O'Brien, L. Sethunya, S. Khumalo

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South Africa is identified as one of the five emerging waste management markets, globally. The waste sector in South Africa influences the areas of energy, water and food at an economic and social level. Recently, South African industries have focused on waste valorization and diversification of the current product offerings in an attempt to reduce industrial waste, target a zero waste-to-landfill initiative and recover energy. South Africa has a number of waste streams including industrial and agricultural biomass, municipal waste and marine waste. Large volumes of agricultural and forestry residues, in particular, are generated which provides significant opportunity for production of bio-based fuels and chemicals. This could directly impact development of a rural economy. One of the largest agricultural industries is the sugar industry, which contributes significantly to the country’s economy and job creation. However, the sugar industry is facing challenges due to fluctuations in sugar prices, increasing competition with low-cost global sugar producers, increasing energy and agricultural input costs, lower consumption and aging facilities. This study is aimed at technology development for the production of various bio-based chemicals using feedstock from the sugar refining process. Various indigenous bacteria and yeast species were assessed for the potential to produce platform chemicals in flask studies and at 30 L fermentation scale. Quantitative analysis of targeted bio-based chemicals was performed using either gas chromatography or high pressure liquid chromatography to assess production yields and techno-economics in order to compare performance to current commercial benchmark processes. The study also creates a decision platform for the research direction that is required for strain development using Industrial Synthetic Biology.

Keywords: bio-based chemicals, biorefinery, industrial synthetic biology, waste valorization

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75 Thermodynamic Analysis and Experimental Study of Agricultural Waste Plasma Processing

Authors: V. E. Messerle, A. B. Ustimenko, O. A. Lavrichshev

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A large amount of manure and its irrational use negatively affect the environment. As compared with biomass fermentation, plasma processing of manure enhances makes it possible to intensify the process of obtaining fuel gas, which consists mainly of synthesis gas (CO + H₂), and increase plant productivity by 150–200 times. This is achieved due to the high temperature in the plasma reactor and a multiple reduction in waste processing time. This paper examines the plasma processing of biomass using the example of dried mixed animal manure (dung with a moisture content of 30%). Characteristic composition of dung, wt.%: Н₂О – 30, С – 29.07, Н – 4.06, О – 32.08, S – 0.26, N – 1.22, P₂O₅ – 0.61, K₂O – 1.47, СаО – 0.86, MgO – 0.37. The thermodynamic code TERRA was used to numerically analyze dung plasma gasification and pyrolysis. Plasma gasification and pyrolysis of dung were analyzed in the temperature range 300–3,000 K and pressure 0.1 MPa for the following thermodynamic systems: 100% dung + 25% air (plasma gasification) and 100% dung + 25% nitrogen (plasma pyrolysis). Calculations were conducted to determine the composition of the gas phase, the degree of carbon gasification, and the specific energy consumption of the processes. At an optimum temperature of 1,500 K, which provides both complete gasification of dung carbon and the maximum yield of combustible components (99.4 vol.% during dung gasification and 99.5 vol.% during pyrolysis), and decomposition of toxic compounds of furan, dioxin, and benz(a)pyrene, the following composition of combustible gas was obtained, vol.%: СО – 29.6, Н₂ – 35.6, СО₂ – 5.7, N₂ – 10.6, H₂O – 17.9 (gasification) and СО – 30.2, Н₂ – 38.3, СО₂ – 4.1, N₂ – 13.3, H₂O – 13.6 (pyrolysis). The specific energy consumption of gasification and pyrolysis of dung at 1,500 K is 1.28 and 1.33 kWh/kg, respectively. An installation with a DC plasma torch with a rated power of 100 kW and a plasma reactor with a dung capacity of 50 kg/h was used for dung processing experiments. The dung was gasified in an air (or nitrogen during pyrolysis) plasma jet, which provided a mass-average temperature in the reactor volume of at least 1,600 K. The organic part of the dung was gasified, and the inorganic part of the waste was melted. For pyrolysis and gasification of dung, the specific energy consumption was 1.5 kWh/kg and 1.4 kWh/kg, respectively. The maximum temperature in the reactor reached 1,887 K. At the outlet of the reactor, a gas of the following composition was obtained, vol.%: СO – 25.9, H₂ – 32.9, СO₂ – 3.5, N₂ – 37.3 (pyrolysis in nitrogen plasma); СO – 32.6, H₂ – 24.1, СO₂ – 5.7, N₂ – 35.8 (air plasma gasification). The specific heat of combustion of the combustible gas formed during pyrolysis and plasma-air gasification of agricultural waste is 10,500 and 10,340 kJ/kg, respectively. Comparison of the integral indicators of dung plasma processing showed satisfactory agreement between the calculation and experiment.

Keywords: agricultural waste, experiment, plasma gasification, thermodynamic calculation

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74 High Level Expression of Fluorinase in Escherichia Coli and Pichia Pastoris

Authors: Lee A. Browne, K. Rumbold

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The first fluorinating enzyme, 5'-fluoro-5'-deoxyadenosine synthase (fluorinase) was isolated from the soil bacterium Streptomyces cattleya. Such an enzyme, with the ability to catalyze a C-F bond, presents great potential as a biocatalyst. Naturally fluorinated compounds are extremely rare in nature. As a result, the number of fluorinases identified remains relatively few. The field of fluorination is almost completely synthetic. However, with the increasing demand for fluorinated organic compounds of commercial value in the agrochemical, pharmaceutical and materials industries, it has become necessary to utilize biologically based methods such as biocatalysts. A key step in this crucial process is the large-scale production of the fluorinase enzyme in considerable quantities for industrial applications. Thus, this study aimed to optimize expression of the fluorinase enzyme in both prokaryotic and eukaryotic expression systems in order to obtain high protein yields. The fluorinase gene was cloned into the pET 41b(+) and pPinkα-HC vectors and used to transform the expression hosts, E.coli BL21(DE3) and Pichia pastoris (PichiaPink™ strains) respectively. Expression trials were conducted to select optimal conditions for expression in both expression systems. Fluorinase catalyses a reaction between S-adenosyl-L-Methionine (SAM) and fluoride ion to produce 5'-fluorodeoxyadenosine (5'FDA) and L-Methionine. The activity of the enzyme was determined using HPLC by measuring the product of the reaction 5'FDA. A gradient mobile phase of 95:5 v/v 50mM potassium phosphate buffer to a final mobile phase containing 80:20 v/v 50mM potassium phosphate buffer and acetonitrile were used. This resulted in the complete separation of SAM and 5’-FDA which eluted at 1.3 minutes and 3.4 minutes respectively. This proved that the fluorinase enzyme was active. Optimising expression of the fluorinase enzyme was successful in both E.coli and PichiaPink™ where high expression levels in both expression systems were achieved. Protein production will be scaled up in PichiaPink™ using fermentation to achieve large-scale protein production. High level expression of protein is essential in biocatalysis for the availability of enzymes for industrial applications.

Keywords: biocatalyst, expression, fluorinase, PichiaPink™

Procedia PDF Downloads 552
73 The Antibacterial and Anticancer Activity of Marine Actinomycete Strain HP411 Isolated in the Northern Coast of Vietnam

Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

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Since the marine environmental conditions are extremely different from the other ones, so that marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411that are effective against wide range of pathogens: both gram-positive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the furture.

Keywords: marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis

Procedia PDF Downloads 419
72 Technico-Economical Study of a Rapeseed Based Biorefinery Using High Voltage Electrical Discharges and Ultrasounds as Pretreatment Technologies

Authors: Marwa Brahim, Nicolas Brosse, Nadia Boussetta, Nabil Grimi, Eugene Vorobiev

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Rapeseed plant is an established product in France which is mainly dedicated to oil production. However, the economic potential of residues from this industry (rapeseed hulls, rapeseed cake, rapeseed straw etc.), has not been fully exploited. Currently, only low-grade applications are found in the market. As a consequence, it was deemed of interest to develop a technological platform aiming to convert rapeseed residues into value- added products. Specifically, a focus is given on the conversion of rapeseed straw into valuable molecules (e.g. lignin, glucose). Existing pretreatment technologies have many drawbacks mainly the production of sugar degradation products that limit the effectiveness of saccharification and fermentation steps in the overall scheme of the lignocellulosic biorefinery. In addition, the viability of fractionation strategies is a challenge in an environmental context increasingly standardized. Hence, the need to find cleaner alternatives with comparable efficiency by implementing physical phenomena that could destabilize the structural integrity of biomass without necessarily using chemical solvents. To meet environmental standards increasingly stringent, the present work aims to study the new pretreatment strategies involving lower consumption of chemicals with an attenuation of the severity of the treatment. These strategies consist on coupling physical treatments either high voltage electrical discharges or ultrasounds to conventional chemical pretreatments (soda and organosolv). Ultrasounds treatment is based on the cavitation phenomenon, and high voltage electrical discharges cause an electrical breakdown accompanied by many secondary phenomena. The choice of process was based on a technological feasibility study taking into account the economic profitability of the whole chain after products valorization. Priority was given to sugars valorization into bioethanol and lignin sale.

Keywords: high voltage electrical discharges, organosolv, pretreatment strategies, rapeseed straw, soda, ultrasounds

Procedia PDF Downloads 362
71 Halloysite Based Adsorbents for Removing Pollutants from Water Reservoirs

Authors: Agata Chelminska, Joanna Goscianska

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The rapid growth of the world’s population and the resulting economic development have had an enormous influence on the environment. Multiple industrial processes generate huge amounts of wastewater containing dangerous substances, most of which are discharged into water bodies. These contaminants include pharmaceuticals and synthetic dyes. Regardless of the presence of wastewater treatment plants, a lot of pollutants cannot be easily eliminated by well-known technologies. Hence, more effective methods of removing resistant chemicals are being developed. Due to cost-effectiveness as well as the availability of a wide range of adsorbents, a large interest in the adsorption process as an alternative way of water purification has been observed. Clay minerals, e.g., halloysite, are one of the most researched natural adsorbents because of their availability, non-toxicity, high specific surface area, porosity, layered structure, and low cost. The negatively charged surface makes them ideal for binding cations and organic compounds. Halloysite can be subjected to modifications which enhance its adsorptive properties. The aim of the presented research was to apply pure and modified halloysite in removing particular pollutants (tetracycline, tartrazine, and phosphates) from aqueous solutions. Halloysite was modified with alcoholic and aqueous solutions of hexadecyltrimethylammonium bromide (CTAB) and urea in different concentrations and subsequently impregnated with lanthanum(III) chloride. Acidic and basic oxygen groups located on the surface of all materials were determined. Moreover, the adsorbents obtained were characterized by X-ray diffraction, low-temperature nitrogen adsorption, scanning, and transmission electron microscopy. The effectiveness of samples in tetracycline, tartrazine, and phosphates adsorption from the liquid phase was then studied in order to determine their potential application in eliminating contaminants from water reservoirs. Modifiers’ employment enabled obtaining materials that possess better adsorption properties, which makes them useful for removing various pollutants from water. Modifying the pure halloysite with CTAB and urea solutions and impregnating LaCl₃ led to the formation of acidic and basic oxygen functional groups on the surface. Their amount increases with an increasing percentage of lanthanum content. The acid-base properties of materials, as well as the type of functional groups that appear on their surface, have a significant influence on their sorption capacities towards antibiotics, dyes, and phosphate(V) anions. The selected contaminants adsorb onto the halloysite studied following the Langmuir type isotherm. The thermodynamic study indicated that the adsorption was a spontaneous and exothermic process. The adsorption equilibrium was rapidly attained after 120 min of contact time. Research showed that synthesized materials based on halloysite may be applied as adsorbents for antibiotics, organic dyes, and PO₄³- ions which are difficult to eliminate.

Keywords: adsorption processes, halloysite, minerals, water reservoirs pollutants

Procedia PDF Downloads 180
70 Exploring the Prebiotic Potential of Glucosamine

Authors: Shilpi Malik, Ramneek Kaur, Archita Gupta, Deepshikha Yadav, Ashwani Mathur, Manisha Singh

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Glucosamine (GS) is the most abundant naturally occurring amino monosaccharide and is normally produced in human body via cellular glucose metabolism. It is regarded as the building block of cartilage matrix and is also an essential component of cartilage matrix repair mechanism. Besides that, it can also be explored for its prebiotic potential as many bacterial species are known to utilize the amino sugar by acquiring them to form peptidoglycans and lipopolysaccharides in the bacterial cell wall. Glucosamine can therefore be considered for its fermentation by bacterial species present in the gut. Current study is focused on exploring the potential of glucosamine as prebiotic. The studies were done to optimize considerable concentration of GS to reach GI tract and being fermented by the complex gut microbiota and food grade GS was added to various Simulated Fluids of Gastro-Intestinal Tract (GIT) such as Simulated Saliva, Gastric Fluid (Fast and Fed State), Colonic fluid, etc. to detect its degradation. Since it was showing increase in microbial growth (CFU) with time, GS was Further, encapsulated to increase its residential time in the gut, which exhibited improved resistance to the simulated Gut conditions. Moreover, prepared microspehres were optimized and characterized for their encapsulation efficiency and toxicity. To further substantiate the prebiotic activity of Glucosamine, studies were also performed to determine the effect of Glucosamine on the known probiotic bacterial species, i.e. Lactobacillus delbrueckii (MTCC 911) and Bifidobacteriumbifidum (MTCC 5398). Culture conditions for glucosamine will be added in MRS media in anaerobic tube at 0.20%, 0.40%, 0.60%, 0.80%, and 1.0%, respectively. MRS media without GS was included in this experiment as the control. All samples were autoclaved at 118° C for 15 min. Active culture was added at 5% (v/v) to each anaerobic tube after cooling to room temperature and incubated at 37° C then determined biomass and pH and viable count at incubation 18h. The experiment was completed in triplicate and the results were presented as Mean ± SE (Standard error).The experimental results are conclusive and suggest Glucosamine to hold prebiotic properties.

Keywords: gastro intestinal tract, microspheres, peptidoglycans, simulated fluid

Procedia PDF Downloads 330
69 Bio Based Agro Textiles

Authors: K. Sakthivel

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With the continuous increase in population worldwide, stress increased among agricultural peoples, so it is necessary to increase the yield of agro-products. But it is not possible to meet fully with the traditionally adopted ways of using pesticides and herbicides. Today, agriculture and horticulture has realized the need of tomorrow and opting for various technologies to get higher overall yield, quality agro-products. Most of today’s synthetic polymers are produced from petrochemical bi-products and are not biodegradable. Persistent polymers generate significant sources of environmental pollution, harming wildlife when they are disposed in nature. The disposal of non degradable plastic bags adversely affects human and wild life. Moreover incineration of plastic waste presents environmental issues as well, since it yields toxic emissions. Material incineration is also limited due to the difficulties to find accurate and economically viable outlets. In addition plastic recycling shows a negative eco balance due to the necessity in nearly all cases to wash the plastic waste as well as the energy consumption during the recycling process phases. As plastics represent a large part of the waste collection at the local regional and national levels institutions are aware of the significant savings that compostable or biodegradable materials would generate. Polylactic acid (PLA), which is one of the most important biocompatible polyesters that are derived from annually renewable biomass such as corn and wheat, has attracted much attention for automotive parts and also can be applied in agro textiles. The manufacturing method of PLA is the ring-opening polymerization of the dimeric cyclic ester of lactic acid, lactide. For the stereo complex PLA, we developed by the four unit processes, fermentation, separation, lactide conversion, and polymerization. Then the polymer is converted into mulching film and applied in agriculture field. PLA agro textiles have better tensile strength, tearing strength and with stand from UV rays than polyester agro textile and polypropylene-based products.

Keywords: biodegradation, environment, mulching film, PLA, technical textiles

Procedia PDF Downloads 386
68 Extracellular Production of the Oncolytic Enzyme, Glutaminase Free L-Asparaginase, from Newly Isolated Streptomyces Olivaceus NEAE-119: Optimization of Culture Conditions Using Response Surface Methodology

Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

Procedia PDF Downloads 365
67 Development and Validation of a Liquid Chromatographic Method for the Quantification of Related Substance in Gentamicin Drug Substances

Authors: Sofiqul Islam, V. Murugan, Prema Kumari, Hari

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Gentamicin is a broad spectrum water-soluble aminoglycoside antibiotics produced by the fermentation process of microorganism known as Micromonospora purpurea. It is widely used for the treatment of infection caused by both gram positive and gram negative bacteria. Gentamicin consists of a mixture of aminoglycoside components like C1, C1a, C2a, and C2. The molecular structure of Gentamicin and its related substances showed that it has lack of presence of chromophore group in the molecule due to which the detection of such components were quite critical and challenging. In this study, a simple Reversed Phase-High Performance Liquid Chromatographic (RP-HPLC) method using ultraviolet (UV) detector was developed and validated for quantification of the related substances present in Gentamicin drug substances. The method was achieved by using Thermo Scientific Hypersil Gold analytical column (150 x 4.6 mm, 5 µm particle size) with isocratic elution composed of methanol: water: glacial acetic acid: sodium hexane sulfonate in the ratio 70:25:5:3 % v/v/v/w as a mobile phase at a flow rate of 0.5 mL/min, column temperature was maintained at 30 °C and detection wavelength of 330 nm. The four components of Gentamicin namely Gentamicin C1, C1a, C2a, and C2 were well separated along with the related substance present in Gentamicin. The Limit of Quantification (LOQ) values were found to be at 0.0075 mg/mL. The accuracy of the method was quite satisfactory in which the % recovery was resulted between 95-105% for the related substances. The correlation coefficient (≥ 0.995) shows the linearity response against concentration over the range of Limit of Quantification (LOQ). Precision studies showed the % Relative Standard Deviation (RSD) values less than 5% for its related substance. The method was validated in accordance with the International Conference of Harmonization (ICH) guideline with various parameters like system suitability, specificity, precision, linearity, accuracy, limit of quantification, and robustness. This proposed method was easy and suitable for use for the quantification of related substances in routine analysis of Gentamicin formulations.

Keywords: reversed phase-high performance liquid chromatographic (RP-HPLC), high performance liquid chromatography, gentamicin, isocratic, ultraviolet

Procedia PDF Downloads 159
66 Comparison of Methods for the Detection of Biofilm Formation in Yeast and Lactic Acid Bacteria Species Isolated from Dairy Products

Authors: Goksen Arik, Mihriban Korukluoglu

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Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Keywords: biofilm, dairy products, lactic acid bacteria, yeast

Procedia PDF Downloads 263
65 Chemical and Biological Studies of Kielmeyera coriacea Mart. (Calophyllaceae) Based on Ethnobotanical Survey of Rural Community from Brazil

Authors: Vanessa G. P. Severino, Eliangela Cristina Candida Costa, Nubia Alves Mariano Teixeira Pires Gomides, Lucilia Kato, Afif Felix Monteiro, Maria Anita Lemos Vasconcelos Ambrosio, Carlos Henrique Gomes Martins

Abstract:

One of the biomes present in Brazil is known as Cerrado, which is a vast tropical savanna ecoregion, particularly in the states of Goiás, Mato Grosso do Sul, Mato Grosso, Tocantins and Minas Gerais. Many species of plants are characterized as endemic and they have therapeutic value for a large part of the population, especially to the rural communities. Given that, the southeastern region of the state of Goiás contains about 21 rural communities, which present a form of organization based on the use of natural resources available. One of these rural communities is named of Coqueiros, where the knowledge about the medicinal plants was very important to this research. Thus, this study focuses on the ethnobotanical survey of this community on the use of Kielmeyera coriacea to treat diseases. From the 37 members interviewed, 76% indicated this species for the treatment of intestinal infection, leukemia, anemia, gastritis, gum pain, toothache, cavity, arthritis, arthrosis, healing, vermifuge, rheumatism, antibiotic, skin problems, mycoses and all kinds of infections. The medicinal properties attributed during the interviews were framed in the body system (disease categories), adapted from ICD 10; thus, 20 indications of use were obtained, among five body systems. Therefore, the root of this species was select to chemical and biological (antioxidant and antimicrobial) studies. From the liquid-liquid extraction of ethanolic extract of root (EER), the hexane (FH), ethyl acetate (FAE), and hydro alcoholic (FHA) fractions were obtained. The chemical profile study of these fractions was performed by LC-MS, identifying major compounds such as δ-tocotrienol, prenylated acylphoroglucinol, 2-hydroxy-1-methoxyxanthone and quercitrin. EER, FH, FAE and FHA were submitted to biological tests. FHA presented the best antioxidant action (EC50 201.53 μg mL-1). EER inhibited the bacterial growth of Streptococcus pyogenes and Pseudomonas aeruginosa, microorganisms associated with rheumatism, at Minimum Inhibitory Concentration (MIC) of 6.25 μg mL-1. In addition, the FH-10 subfraction, obtained from FH fractionation, presented MIC of 1.56 μg mL-1 against S. pneumoniae; EER also inhibited the fungus Candida glabrata (MIC 7.81 μg mL- 1). The FAE-4.7.3 fraction, from the fractionation of FAE, presented MIC of 200 μg mL-1 against Lactobacillus casei, which is one of the causes of caries and oral infections. By the correlation of the chemical and biological data, it is possible to note that the FAE-4.7.3 and FH-10 are constituted 4-hydroxy-2,3-methylenedioxy xanthone, 3-hydroxy-1,2-dimethoxy xanthone, lupeol, prenylated acylphoroglucinol and quercitrin, which could be associated with the biological potential found. Therefore, this study provides an important basis for further investigations regarding the compounds present in the active fractions of K. coriacea, which will permit the establishment of a correlation between ethnobotanical survey and bioactivity.

Keywords: biological activity, ethnobotanical survey, Kielmeyera coriacea Mart., LC-MS profile

Procedia PDF Downloads 140
64 Process of Production of an Artisanal Brewery in a City in the North of the State of Mato Grosso, Brazil

Authors: Ana Paula S. Horodenski, Priscila Pelegrini, Salli Baggenstoss

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The brewing industry with artisanal concepts seeks to serve a specific market, with diversified production that has been gaining ground in the national environment, also in the Amazon region. This growth is due to the more demanding consumer, with a diversified taste that wants to try new types of beer, enjoying products with new aromas, flavors, as a differential of what is so widely spread through the big industrial brands. Thus, through qualitative research methods, the study aimed to investigate how is the process of managing the production of a craft brewery in a city in the northern State of Mato Grosso (BRAZIL), providing knowledge of production processes and strategies in the industry. With the efficient use of resources, it is possible to obtain the necessary quality and provide better performance and differentiation of the company, besides analyzing the best management model. The research is descriptive with a qualitative approach through a case study. For the data collection, a semi-structured interview was elaborated, composed of the areas: microbrewery characterization, artisan beer production process, and the company supply chain management. Also, production processes were observed during technical visits. With the study, it was verified that the artisan brewery researched develops preventive maintenance strategies with the inputs, machines, and equipment, so that the quality of the product and the production process are achieved. It was observed that the distance from the supplying centers makes the management of processes and the supply chain be carried out with a longer planning time so that the delivery of the final product is satisfactory. The production process of the brewery is composed of machines and equipment that allows the control and quality of the product, which the manager states that for the productive capacity of the industry and its consumer market, the available equipment meets the demand. This study also contributes to highlight one of the challenges for the development of small breweries in front of the market giants, that is, the legislation, which fits the microbreweries as producers of alcoholic beverages. This makes the micro and small business segment to be taxed as a major, who has advantages in purchasing large batches of raw materials and tax incentives because they are large employers and tax pickers. It was possible to observe that the supply chain management system relies on spreadsheets and notes that are done manually, which could be simplified with a computer program to streamline procedures and reduce risks and failures of the manual process. In relation to the control of waste and effluents affected by the industry is outsourced and meets the needs. Finally, the results showed that the industry uses preventive maintenance as a productive strategy, which allows better conditions for the production and quality of artisanal beer. The quality is directly related to the satisfaction of the final consumer, being prized and performed throughout the production process, with the selection of better inputs, the effectiveness of the production processes and the relationship with the commercial partners.

Keywords: artisanal brewery, production management, production processes, supply chain

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63 Evaluating the Performance of Organic, Inorganic and Liquid Sheep Manure on Growth, Yield and Nutritive Value of Hybrid Napier CO-3

Authors: F. A. M. Safwan, H. N. N. Dilrukshi, P. U. S. Peiris

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Less availability of high quality green forages leads to low productivity of national dairy herd of Sri Lanka. Growing grass and fodder to suit the production system is an efficient and economical solution for this problem. CO-3 is placed in a higher category, especially on tillering capacity, green forage yield, regeneration capacity, leaf to stem ratio, high crude protein content, resistance to pests and diseases and free from adverse factors along with other fodder varieties grown within the country. An experiment was designed to determine the effect of organic sheep manure, inorganic fertilizers and liquid sheep manure on growth, yield and nutritive value of CO-3. The study was consisted with three treatments; sheep manure (T1), recommended inorganic fertilizers (T2) and liquid sheep manure (T3) which was prepared using bucket fermentation method and each treatment was consisted with three replicates and those were assigned randomly. First harvest was obtained after 40 days of plant establishment and number of leaves (NL), leaf area (LA), tillering capacity (TC), fresh weight (FW) and dry weight (DW) were recorded and second harvest was obtained after 30 days of first harvest and same set of data were recorded. SPSS 16 software was used for data analysis. For proximate analysis AOAC, 2000 standard methods were used. Results revealed that the plants treated with T1 recorded highest NL, LA, TC, FW and DW and were statistically significant at first and second harvest of CO-3 (p˂ 0.05) and it was found that T1 was statistically significant from T2 and T3. Although T3 was recorded higher than the T2 in almost all growth parameters; it was not statistically significant (p ˃0.05). In addition, the crude protein content was recorded highest in T1 with the value of 18.33±1.61 and was lowest in T2 with the value of 10.82±1.14 and was statistically significant (p˂ 0.05). Apart from this, other proximate composition crude fiber, crude fat, ash, moisture content and dry matter were not statistically significant between treatments (p ˃0.05). In accordance with the results, it was found that the organic fertilizer is the best fertilizer for CO-3 in terms of growth parameters and crude protein content.

Keywords: fertilizer, growth parameters, Hybrid Napier CO-3, proximate composition

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62 Compositional Assessment of Fermented Rice Bran and Rice Bran Oil and Their Effect on High Fat Diet Induced Animal Model

Authors: Muhammad Ali Siddiquee, Md. Alauddin, Md. Omar Faruque, Zakir Hossain Howlader, Mohammad Asaduzzaman

Abstract:

Rice bran (RB) and rice bran oil (RBO) are explored as prominent food components worldwide. In this study, fermented rice bran (FRB) was produced by employing edible gram-positive bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus, and Bifidobacterium bifidum) at 125 x 10⁵ spore g⁻¹ of rice bran, and investigated to evaluate nutritional quality. The crude rice bran oil (CRBO) was extracted from RB, and its quality was also investigated compared to market-available rice bran oil (MRBO) in Bangladesh. We found that fermentation of rice bran with lactic acid bacteria increased total proteins (29.52%), fat (5.38%), ash (48.47%), crude fiber (38.96%), and moisture (61.04%) and reduced the carbohydrate content (36.61%). We also found that essential amino acids (methionine, tryptophan, threonine, valine, leucine, lysine, histidine, and phenylalanine) and non-essential amino acids (alanine, aspartate, glycine, glutamine, proline, serine, and tyrosine) were increased in FRB except methionine and proline. Moreover, total phenolic content, tannin content, flavonoid content, and antioxidant activity were increased in FRB. The RBO analysis showed that γ-oryzanol content (10.00mg/g) was found in CRBO compared to MRBO (ranging from 7.40 to 12.70 mg/g) and Vitamin-E content 0.20% was found higher in CRBO compared to MRBO (ranging 0.097 to 0.12%). The total saturated (25.16%) and total unsaturated fatty acids (74.44%) were found in CRBO, whereas MRBO contained total saturated (22.08 to 24.13%) and total unsaturated fatty acids (71.91 to 83.29%), respectively. The physiochemical parameters were found satisfactory in all samples except acid value and peroxide value higher in CRBO. Finally, animal experiments showed that FRB and CRBO reduce the body weight, glucose, and lipid profile in high-fat diet-induced animal models. Thus, FRB and RBO could be value-added food supplements for human health.

Keywords: fermented rice bran, crude rice bran oil, amino acids, proximate composition, gamma-oryzanol, fatty acids, heavy metals, physiochemical parameters

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61 In Vitro Digestibility of Grains and Straw of Seventeen Ecotypes of Bitter Vetch (Vicia ervilia) in the North of Morocco

Authors: Boukrouh Soumaya, Cabaraux Jean-François, Avril Claire, Noutfia Ali, Chentouf Mouad

Abstract:

The introduction of marginal leguminous forage species in the diet of ruminants are of great importance. Bitter vetch is a good source of proteins, highly resistant against drought and poor soil conditions. Accordingly; two years field trials (2018/2019 and 2019-2020) were conducted to determine the digestibility of straw and grains of 17 promising bitter vetch ecotypes(Vicia ervilia) in the north of Morocco. In vitro dry and organic matter digestibility, gas production, and kinetics of fermentation of grains and straw were evaluated using gas production technique, pepsin-cellulase enzymatic digestibility of DM (CDDM)and OM (CDOM), as well as protease enzymatic CP degradation (CPD) and in vitro true digestibility, were performed using DAISYII Incubator. In vitro digestibility was performed using gas production method of (Menke et al., 1979) improved by Menke and Steingass (1988). Samples were incubated in glass syringes that contained rumen fluid and incubation solution that conserved in water bath in 39°C during 72 hours. Gas production was recorded after 2, 4, 8, 12, 24, 48, and 72 hours. Studied digestibility parameters were dry and organic matter digestibility, microbial biomass production, partitioning factor, and volatile fatty acids. Enzymatic dry matter digestibility was different (p < 0.05) among grains and straw for all ecotypes. It varied from 804.1 to 957.7 g/kg DM and 270.4 to 412.3 g/kg DM for grains and straw, respectively. Metabolizable energy varied between 11.7 to 14.3 MJ/kg DM and 2.6 to 5.0 MJ/kg DM for grains and straw, respectively. Potential gas production (A), the rate constants (c and d), and lag times of grains and straws from different bitter vetch ecotypes were different (p > 0.05). The results emphasized that in any evaluation of bitter vetch ecotypes, where straw of this legume seed is used as an animal feed, not only seed yield but also yield and quality of straw should be taken into consideration, particularly in areas where straw from this legume is considered as an important feedstuff for ruminants. Enzymatic digestibility was lower than in vitro digestibility by gaz production and by the DAISYII method because rumen fluid contains bacteria than increase digestibility. There was no difference between in vitro digestibility by gaz production and the DAISY II method. The DAISY II method can be used to increase labor efficiency in the in vitro DM digestibility analysis if gaz production is not necessary for analysis.

Keywords: bitter vetch, grains, straw, ecotype, in vitro digestibility, gaz production, enzymatic digestibility

Procedia PDF Downloads 177