Search results for: Rumen fermentation

Authors: Wai Prathumpai, Pranee Rachtawee, Sutamat Khajeeram, Pariya Na Nakorn

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The  β-glucan produced by Ophiocordyceps dipterigena BCC 2073 is a (1, 3)-β-D-glucan with highly branching O-6-linkedside chains that is resistant to acid hydrolysis (by hydrochloric acid and porcine pancreatic alpha-amylase). This β-glucan can be utilized as a prebiotic due to its advantageous structural and biological properties. The effects of using this β-glucan as the sole carbon source for the in vitro growth of two probiotic bacteria (L. acidophilus BCC 13938 and B. animalis ATCC 25527) were investigated. Compared with the effect of using 1% glucose or fructo-oligosaccharide (FOS) as the sole carbon source, using 1% β-glucan for this purpose showed that this prebiotic supported and stimulated the growth of both types of probiotic bacteria and induced them to produce the highest levels of metabolites during their growth. The highest levels of lactic and acetic acid, 10.04 g·L^-1 and 2.82 g·L^-1, respectively, were observed at 2 h of cultivation using glucose as the sole carbon source. Furthermore, the fermentation broth obtained using 1% β-glucan as the sole carbon source had greater antibacterial activity against selected pathogenic bacteria (B. subtilis TISTR 008, E. coli TISTR 780, and S. typhimurium TISTR 292) than did the broths prepared using glucose or FOS as the sole carbon source. The fermentation broth obtained by growing L. acidophilus BCC 13938 in the presence of β-glucan inhibited the growth of B. subtilis TISTR 008 by more than 70% and inhibited the growth of both S. typhimurium TISTR 292 and E. coli TISTR 780 by more than 90%. In conclusion, O. dipterigena BCC 2073 is a potential source of a β-glucan prebiotic that could be used for commercial production in the near future.

Keywords: beta-glucan, Ophiocordyceps dipterigena, prebiotic, probiotic, antimicrobial

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Authors: Nurcan Cetinkaya

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The aim of this paper is to bring about the utilization of Juncus acutus as an alternative roughage resource in ruminant nutrition. In Turkey, JA is prevailing plant of the natural grassland in Kizilirmak Delta, Samsun. Crude nutrient values such as crude protein (CP), ether extract (EE), organic matter (OM), neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin(ADL) including antioxidant activity, total phenolic and flavonoid compounds, total organic matter digestibility (OMD) and metabolisable energy (ME) values of Juncus acutus stem, seed, and also its mixture with maize silage were estimated. and published. Furthermore, the effects of JA over rumen cellulolitic bacteria were studied. The obtained results from different studies conducted on JA by our team show that Juncus acutus may be a new roughage source in ruminant nutrition.

Keywords: antioxidant activity, cellulolytic bacteria, Juncus acutus, organic matter digestibility

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Authors: Idan Chiyanzu, Maruping Mangena

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Crude glycerol, a major by-product from the transesterification of triacylglycerides with alcohol to biodiesel, is known to have a broad range of applications. For example, its bioconversion can afford a wide range of chemicals including alcohols, organic acids, hydrogen, solvents and intermediate compounds. In bacteria, the 2-oxoglutarate dioxygenase (2-OGD) enzymes are widely found among the Pseudomonas syringae species and have been recognized with an emerging importance in ethylene formation. However, the use of optimized enzyme function in recombinant systems for crude glycerol conversion to ethylene is still not been reported. The present study investigated the production of ethylene from crude glycerol using engineered E. coli MG1655 and JM109 strains. Ethylene production with an optimized expression system for 2-OGD in E. coli using a codon optimized construct of the ethylene-forming gene was studied. The codon-optimization resulted in a 20-fold increase of protein production and thus an enhanced production of the ethylene gas. For a reliable bioreactor performance, the effect of temperature, fermentation time, pH, substrate concentration, the concentration of methanol, concentration of potassium hydroxide and media supplements on ethylene yield was investigated. The results demonstrate that the recombinant enzyme can be used for future studies to exploit the conversion of low-priced crude glycerol into advanced value products like light olefins, and tools including recombineering techniques for DNA, molecular biology, and bioengineering can be used to allowing unlimited the production of ethylene directly from the fermentation of crude glycerol. It can be concluded that recombinant E.coli production systems represent significantly secure, renewable and environmentally safe alternative to thermochemical approach to ethylene production.

Keywords: crude glycerol, bioethylene, recombinant E. coli, optimization

Procedia PDF Downloads 259

Authors: Joyce Cruz Ferraz Dutra, Marcele Fonseca Passos, Rubens Maciel Filho, Douglas Fernandes Barbin, Gustavo Mockaitis

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The search for alternatives to control hunger in the world, generated a major environmental problem. Intensive systems of fish production can cause an imbalance in the aquatic environment, triggering the phenomenon of eutrophication. Currently, there are many forms of growth control aquatic plants, such as mechanical withdrawal, however some difficulties arise for their final destination. The Egeria densa is a species of submerged aquatic macrophyte-rich in cellulose and low concentrations of lignin. By applying the concept of second generation energy, which uses lignocellulose for energy production, the reuse of these aquatic macrophytes (Egeria densa) in the biofuels production can turn an interesting alternative. In order to make lignocellulose sugars available for effective fermentation, it is important to use pre-treatments in order to separate the components and modify the structure of the cellulose and thus facilitate the attack of the microorganisms responsible for the fermentation. Therefore, the objective of this research work was to evaluate the structural and morphological transformations occurring in the biomass of aquatic macrophytes (E.densa) submitted to a thermal pretreatment. The samples were collected in an intensive fish growing farm, in the low São Francisco dam, in the northeastern region of Brazil. After collection, the samples were dried in a 65 0C ventilation oven and milled in a 5mm micron knife mill. A duplicate assay was carried, comparing the in natural biomass with the pretreated biomass with heat (MT). The sample (MT) was submitted to an autoclave with a temperature of 1210C and a pressure of 1.1 atm, for 30 minutes. After this procedure, the biomass was characterized in terms of degree of crystallinity and morphology, using X-ray diffraction (XRD) techniques and scanning electron microscopy (SEM), respectively. The results showed that there was a decrease of 11% in the crystallinity index (% CI) of the pretreated biomass, leading to the structural modification in the cellulose and greater presence of amorphous structures. Increases in porosity and surface roughness of the samples were also observed. These results suggest that biomass may become more accessible to the hydrolytic enzymes of fermenting microorganisms. Therefore, the morphological transformations caused by the thermal pretreatment may be favorable for a subsequent fermentation and, consequently, a higher yield of biofuels. Thus, the use of thermally pretreated aquatic macrophytes (E.densa) can be an environmentally, financially and socially sustainable alternative. In addition, it represents a measure of control for the aquatic environment, which can generate income (biogas production) and maintenance of fish farming activities in local communities.

Keywords: aquatics macrophyte, biofuels, crystallinity, morphology, pretreatment thermal

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Authors: Marcin Zielinski, Marcin Debowski, Paulina Rusanowska, Magda Dudek

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As a result of sonification, the destruction of complex biomass structures results in an increase in the biogas yield from the conditioned material. First, the amount of organic matter released into the solution due to disintegration was determined. This parameter was determined by changes in the carbon content in liquid phase of the conditioned substrate. The amount of carbon in the liquid phase increased with the prolongation of the sonication time to 16 min. Further increase in the duration of sonication did not cause a statistically significant increase in the amount of organic carbon in the liquid phase. The disintegrated material was then used for respirometric measurements for determination of the impact of the conditioning process used on methane fermentation effectiveness. The relationship between the amount of energy introduced into the lignocellulosic substrate and the amount of biogas produced has been demonstrated. Statistically significant increase in the amount of biogas was observed until sonication of 16 min. Further increase in energy in the conditioning process did not significantly increase the production of biogas from the treated substrate. The biogas production from the conditioned substrate was 17% higher than from the reference biomass at that time. The ultrasonic disintegration method did not significantly affect the observed biogas composition. In all series, the methane content in the produced biogas from the conditioned substrate was similar to that obtained with the raw substrate sample (51.1%). Another method of substrate conditioning was hydrothermal depolymerization. This method consists in application of increased temperature and pressure to substrate. These phenomena destroy the structure of the processed material, the release of organic compounds to the solution, which should lead to increase the amount of produced biogas from such treated biomass. The hydrothermal depolymerization was conducted using an innovative microwave heating method. Control measurements were performed using conventional heating. The obtained results indicate the relationship between depolymerization temperature and the amount of biogas. Statistically significant value of the biogas production coefficients increased as the depolymerization temperature increased to 150°C. Further raising the depolymerization temperature to 180°C did not significantly increase the amount of produced biogas in the respirometric tests. As a result of the hydrothermal depolymerization obtained using microwave at 150°C for 20 min, the rate of biogas production from the Sida silage was 780 L/kg VS, which accounted for nearly 50% increase compared to 370 L/kg VS obtained from the same silage but not depolymerised. The study showed that by microwave heating it is possible to effectively depolymerized substrate. Significant differences occurred especially in the temperature range of 130-150ºC. The pre-treatment of Sida hermaphrodita silage (biogas substrate) did not significantly affect the quality of the biogas produced. The methane concentration was about 51.5% on average. The study was carried out in the framework of the project under program BIOSTRATEG funded by the National Centre for Research and Development No. 1/270745/2/NCBR/2015 'Dietary, power, and economic potential of Sida hermaphrodita cultivation on fallow land'.

Keywords: disintegration, biogas, methane fermentation, Virginia fanpetals, biomass

Procedia PDF Downloads 269

Authors: Zhanara Suleimenova, Raushan Blieva, Aigerim Zhakipbekova, Inkar Tapenbayeva, Zhanar Narmuratova

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Green processes are based on innovative technologies that do not negatively affect the environment. Industrial enzymes originated from biological systems can effectively contribute to sustainable development through being isolated from microorganisms which are fermented using primarily renewable resources. Many widespread microorganisms secrete a significant amount of biocatalysts into the environment, which greatly facilitates the task of their isolation and purification. The ability to control the enzyme production through the regulation of their biosynthesis and the selection of nutrient media and cultivation conditions allows not only to increase the yield of enzymes but also to obtain enzymes with certain properties. In this regard, large potentialities are embedded in immobilized cells. Enzyme production technology in a secreted active form enabling industrial application on an economically feasible scale has been developed. This method is based on the immobilization of enzyme producers on a solid career. Immobilizing has a range of advantages: decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, the ability of various enzymes' simultaneous production, etc. Design of proposed equipment gives the opportunity to increase the activity of immobilized cell culture filtrate comparing to free cells, growing in periodic culture conditions. Such technology allows giving a 10-times raise in culture productivity, to prolong the process of fungi cultivation and periods of active culture liquid generation. Also, it gives the way to improve the quality of filtrates (to make them more clear) and exclude time-consuming processes of recharging fermentative vials, that require manual removing of mycelium.

Keywords: industrial enzymes, immobilization, submerged fermentation, microscopic fungi

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Authors: Muhammad Zainuddin Arriafdi, Hamudah Hakimah Abdullah, Mohd Helmi Sani, Wan Azlina Ahmad, Muhd Nazrul Hisham Zainal Alam

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The biotechnology process development demands numerous experimental works. In laboratory environment, this is typically carried out using a shake flask platform. This paper presents the design and fabrication of a miniature bioreactor system as an alternative research tool for bioprocessing. The working volume of the reactor is 100 ml, and it is made of plastic. The main features of the reactor included stirring control, temperature control via the electrical heater, aeration strategy through a miniature air compressor, and online optical cell density (OD) sensing. All sensors and actuators integrated into the reactor was controlled using an Arduino microcontroller platform. In order to demonstrate the functionality of such miniature bioreactor concept, series of batch Saccharomyces cerevisiae fermentation experiments were performed under various glucose concentrations. Results attained from the fermentation experiments were utilized to solve the Monod equation constants, namely the saturation constant, Ks, and cells maximum growth rate, μmax as to further highlight the usefulness of the device. The mixing capacity of the reactor was also evaluated. It was found that the results attained from the miniature bioreactor prototype were comparable to results achieved using a shake flask. The unique features of the device as compared to shake flask platform is that the reactor mixing condition is much more comparable to a lab-scale bioreactor setup. The prototype is also integrated with an online OD sensor, and as such, no sampling was needed to monitor the progress of the reaction performed. Operating cost and medium consumption are also low and thus, making it much more economical to be utilized for biotechnology process development compared to lab-scale bioreactors.

Keywords: biotechnology, miniature bioreactor, research tools, Saccharomyces cerevisiae

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Authors: Sidra Pervez, Afsheen Aman, Shah Ali Ul Qader

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The importance of attaining an optimum performance of an enzyme is often a question of devising an effective method for its immobilization. Cross-linked enzyme aggregate (CLEAs) is a new approach for immobilization of enzymes using carrier free strategy. This method is exquisitely simple (involving precipitation of the enzyme from aqueous buffer followed by cross-linking of the resulting physical aggregates of enzyme molecules) and amenable to rapid optimization. Among many industrial enzymes, amyloglucosidase is an important amylolytic enzyme that hydrolyzes alpha (1→4) and alpha (1→6) glycosidic bonds in starch molecule and produce glucose as a sole end product. Glucose liberated by amyloglucosidase can be used for the production of ethanol and glucose syrups. Besides this amyloglucosidase can be widely used in various food and pharmaceuticals industries. For production of amyloglucosidase on commercial scale, filamentous fungi of genera Aspergillus are mostly used because they secrete large amount of enzymes extracellularly. The current investigation was based on isolation and identification of filamentous fungi from genus Aspergillus for the production of amyloglucosidase in submerged fermentation and optimization of cultivation parameters for starch saccharification. Natural isolates were identified as Aspergillus niger KIBGE-IB36, Aspergillus fumigatus KIBGE-IB33, Aspergillus flavus KIBGE-IB34 and Aspergillus terreus KIBGE-IB35 on taxonomical basis and 18S rDNA analysis and their sequence were submitted to GenBank. Among them, Aspergillus fumigatus KIBGE-IB33 was selected on the basis of maximum enzyme production. After optimization of fermentation conditions enzyme was immobilized on CLEA. Different parameters were optimized for maximum immobilization of amyloglucosidase. Data of enzyme stability (thermal and Storage) and reusability suggested the applicability of immobilized amyloglucosidase for continuous saccharification of starch in industrial processes.

Keywords: aspergillus, immobilization, industrial processes, starch saccharification

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Authors: Strahinja Kovacevic, Milica Karadzic Banjac, Jasmina Vitas, Stefan Vukmanovic, Radomir Malbasa, Lidija Jevric, Sanja Podunavac-Kuzmanovic

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The present study deals with chemometric regression analysis of quality parameters and the radical scavenging ability of kombucha fermented kefir-like products obtained with winter savory (WS), peppermint (P), stinging nettle (SN) and wild thyme tea (WT) kombucha inoculums. Each analyzed sample was described by milk fat content (MF, %), total unsaturated fatty acids content (TUFA, %), monounsaturated fatty acids content (MUFA, %), polyunsaturated fatty acids content (PUFA, %), the ability of free radicals scavenging (RSA Dₚₚₕ, % and RSA.ₒₕ, %) and pH values measured after each hour from the start until the end of fermentation. The aim of the conducted regression analysis was to establish chemometric models which can predict the radical scavenging ability (RSA Dₚₚₕ, % and RSA.ₒₕ, %) of the samples by correlating it with the MF, TUFA, MUFA, PUFA and the pH value at the beginning, in the middle and at the end of fermentation process which lasted between 11 and 17 hours, until pH value of 4.5 was reached. The analysis was carried out applying univariate linear (ULR) and multiple linear regression (MLR) methods on the raw data and the data standardized by the min-max normalization method. The obtained models were characterized by very limited prediction power (poor cross-validation parameters) and weak statistical characteristics. Based on the conducted analysis it can be concluded that the resulting radical scavenging ability cannot be precisely predicted only on the basis of MF, TUFA, MUFA, PUFA content, and pH values, however, other quality parameters should be considered and included in the further modeling. This study is based upon work from project: Kombucha beverages production using alternative substrates from the territory of the Autonomous Province of Vojvodina, 142-451-2400/2019-03, supported by Provincial Secretariat for Higher Education and Scientific Research of AP Vojvodina.

Keywords: chemometrics, regression analysis, kombucha, quality control

Procedia PDF Downloads 112

Authors: Sanjiv Kumar Soni, Chetna Janveja

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The approach of utilizing zero cost kitchen waste residues for growing suitable strains of fungi for the induction of a cocktail of hydrolytic enzymes and ethanol generation has been validated in the present study with the objective of developing an indigenous biorefinery for low cost bioethanol production with the generation of zero waste. Solid state fermentation has been carried out to evaluate the potential of various steam pretreated kitchen waste residues as substrates for the co-production of multiple carbohydrases including cellulases, hemicellulases, pectinase and amylases by a locally isolated strain of Aspergillus niger C-5. Of all the residues, potato peels induced the maximum yields of all the enzyme components corresponding to 64.0±1.92 IU of CMCase, 17.0±0.54 IU of FPase , 42.8±1.28 IU of β-glucosidase, 990.0±28.90 IU of xylanase, 53.2±2.12 IU of mannanase, 126.0±3.72 IU of pectinase, 31500.0±375.78 IU of α-amylase and 488.8±9.82 IU of glucoamylase/g dry substrate respectively. Saccharification of various kitchen refuse residues using inhouse produced crude enzyme cocktail resulted in the release of 610±10.56, 570±8.89, 435±6.54, 475±4.56, 445±4.27, 385±4.49, 370±6.89, 490±10.45 mg of total reducing sugars/g of dried potato peels, orange peels, pineapple peels, mausami peels, onion peels, banana stalks, pea pods and composite mixture respectively revealing carbohydrate conversion efficiencies in the range of 97.0-99.4%. After fermentation of released hexoses by Saccharomyces cerevisae, ethanol yields ranging from 80-262 mL/ kg of dry residues were obtained. The study has successfully evaluated the valorization of kitchen garbage, a highly biodegradable component in Municipal Solid Waste by using it as a substrate for the in-house co-production of multiple carbohydrases and employing the steam treated residues as a feed stock for bioethanol production. Such valorization of kitchen garbage may reduce the level of Municipal Solid Waste going into land-fills thus lowering the emissions of greenhouse gases. Moreover, the solid residue left after the bioconversion may be used as a biofertilizer for improving the fertility of the soils.

Keywords: kitchen waste, bioethanol, solid waste, bioconversion, waste management

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Authors: Lucia Steenkamp, Chris V. D. Westhuyzen, Kgama Mathiba

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Ambergris, and more specifically its oxidation product (–)-ambrafuran, is a scarce, valuable, and sought-after perfumery ingredient. The material is used as a fixative agent to stabilise perfumes in formulations by reducing the evaporation rate of volatile substances. Ambergris is a metabolic product of the sperm whale (Physeter macrocephatus L.), resulting from intestinal irritation. Chemically, (–)-ambrafuran is produced from the natural product sclareol in eight synthetic steps – in the process using harsh and often toxic chemicals to do so. An overall yield of no more than 76% can be achieved in some routes, but generally, this is lower. A new 'green' route has been developed in our laboratory in which sclareol, extracted from the Clary sage plant, is converted to (–)-ambrafuran in two steps with an overall yield in excess of 80%. The first step uses a microorganism, Hyphozyma roseoniger, to bioconvert sclareol to an intermediate diol using substrate concentrations up to 50g/L. The yield varies between 90 and 67% depending on the substrate concentration used. The purity of the diol product is 95%, and the diol is used without further purification in the next step. The intermediate diol is then cyclodehydrated to the final product (–)-ambrafuran using a zeolite, which is not harmful to the environment and is readily recycled. The yield of the product is 96%, and following a single recrystallization, the purity of the product is > 99.5%. A preliminary LC-MS study of the bioconversion identified several intermediates produced in the fermentation broth under oxygen-restricted conditions. Initially, a short-lived ketone is produced in equilibrium with a more stable pyranol, a key intermediate in the process. The latter is oxidised under Norrish type I cleavage conditions to yield an acetate, which is hydrolysed either chemically or under lipase action to afford the primary fermentation product, an intermediate diol. All the intermediates identified point to the likely CYP450 action as the key enzyme(s) in the mechanism. This invention is an exceptional example of how the power of biocatalysis, combined with a mild, benign chemical step, can be deployed to replace a total chemical synthesis of a specific chiral antipode of a commercially relevant material.

Keywords: ambrafuran, biocatalysis, fragrance, microorganism

Procedia PDF Downloads 174

Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

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Authors: Kailas L. Wasewar

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Biobutanol has been considered as a potential and alternative biofuel relative to the most popular biodiesel and bioethanol. End product toxicity is the major problems in commercialization of fermentation based process which can be reduce to some possible extent by removing biobutanol simultaneously. Several techniques have been investigated for removing butanol from fermentation broth such as stripping, adsorption, liquid–liquid extraction, pervaporation, and membrane solvent extraction. Liquid–liquid extraction can be performed with high selectivity and is possible to carry out inside the fermenter. Conventional solvents have few drawbacks including toxicity, loss of solvent, high cost etc. Hence alternative solvents must be explored for the same. Room temperature ionic liquids (RTILs) composed entirely of ions are liquid at room temperature having negligible vapor pressure, non-flammability, and tunable physiochemical properties for a particular application which term them as “designer solvents”. Ionic liquids (ILs) have recently gained much attention as alternatives for organic solvents in many processes. In particular, ILs have been used as alternative solvents for liquid–liquid extraction. Their negligible vapor pressure allows the extracted products to be separated from ILs by conventional low pressure distillation with the potential for saving energy. Morpholinium, imidazolium, ammonium, phosphonium etc. based ionic liquids have been employed for the separation biobutanol. In present chapter, basic concepts of ionic liquids and application in separation have been presented. Further, type of ionic liquids including, conventional, functionalized, polymeric, supported membrane, and other ionic liquids have been explored. Also the effect of various performance parameters on separation of biobutanol by ionic liquids have been discussed and compared for different cation and anion based ionic liquids. The typical methodology for investigation have been adopted such as contacting the equal amount of biobutanol and ionic liquids for a specific time say, 30 minutes to confirm the equilibrium. Further, biobutanol phase were analyzed using GC to know the concentration of biobutanol and material balance were used to find the concentration in ionic liquid.

Keywords: biobutanol, separation, ionic liquids, sustainability, biorefinery, waste biomass

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Authors: Anubha Kaushik, Raman Preet

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Distillery wastewater in the form of spent wash is a complex and strong industrial effluent, with high load of organic pollutants that may deplete dissolved oxygen on being discharged into aquatic systems and contaminate groundwater by leaching of pollutants, while untreated spent wash disposed on land acidifies the soil. Stringent legislative measures have therefore been framed in different countries for discharge standards of distillery effluent. Utilising the organic pollutants present in various types of wastes as food by mixed microbial populations is emerging as an eco-friendly approach in the recent years, in which complex organic matter is converted into simpler forms, and simultaneously useful gases are produced as renewable and clean energy sources. In the present study, wastewater from a rice bran based distillery has been used as the substrate in a dark fermenter, and native microbial consortium from the digester sludge has been used as the inoculum to treat the wastewater and produce hydrogen. After optimising the operational conditions in batch reactors, sequential batch mode and continuous flow stirred tank reactors were used to study the best operational conditions for enhanced and sustained hydrogen production and removal of pollutants. Since the rate of hydrogen production by the microbial consortium during dark fermentation is influenced by concentration of organic matter, pH and temperature, these operational conditions were optimised in batch mode studies. Maximum hydrogen production rate (347.87ml/L/d) was attained in 32h dark fermentation while a good proportion of COD also got removed from the wastewater. Slightly acidic initial pH seemed to favor biohydrogen production. In continuous stirred tank reactor, high H₂ production from distillery wastewater was obtained from a relatively shorter substrate retention time (SRT) of 48h and a moderate organic loading rate (OLR) of 172 g/l/d COD.

Keywords: distillery wastewater, hydrogen, microbial consortium, organic pollution, sludge

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Authors: Bahiru Tsegaye, Chandrajit Balomajumder, Partha Roy

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The limited supply and related negative environmental consequence of fossil fuels are driving researcher for finding sustainable sources of energy. Lignocellulose biomass like sorghum straw is considered as among cheap, renewable and abundantly available sources of energy. However, lignocellulose biomass conversion to bioenergy like bioethanol is hindered due to the reluctant nature of lignin in the biomass. Therefore, removal of lignin is a vital step for lignocellulose conversion to renewable energy. The aim of this study is to optimize microwave pretreatment conditions using design expert software to remove lignin and to release maximum possible polysaccharides from sorghum straw for efficient hydrolysis and fermentation process. Sodium hydroxide concentration between 0.5-1.5%, v/v, pretreatment time from 5-25 minutes and pretreatment temperature from 120-2000C were considered to depolymerize sorghum straw. The effect of pretreatment was studied by analyzing the compositional changes before and after pretreatments following renewable energy laboratory procedure. Analysis of variance (ANOVA) was used to test the significance of the model used for optimization. About 32.8%-48.27% of hemicellulose solubilization, 53% -82.62% of cellulose release, and 49.25% to 78.29% lignin solubilization were observed during microwave pretreatment. Pretreatment for 10 minutes with alkali concentration of 1.5% and temperature of 1400C released maximum cellulose and lignin. At this optimal condition, maximum of 82.62% of cellulose release and 78.29% of lignin removal was achieved. Sorghum straw at optimal pretreatment condition was subjected to enzymatic hydrolysis and fermentation. The efficiency of hydrolysis was measured by analyzing reducing sugars by 3, 5 dinitrisylicylic acid method. Reducing sugars of about 619 mg/g of sorghum straw were obtained after enzymatic hydrolysis. This study showed a significant amount of lignin removal and cellulose release at optimal condition. This enhances the yield of reducing sugars as well as ethanol yield. The study demonstrates the potential of microwave pretreatments for enhancing bioethanol yield from sorghum straw.

Keywords: cellulose, hydrolysis, lignocellulose, optimization

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Authors: Elias L. Souza, Noeli Sellin, Cintia Marangoni, Ozair Souza

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Among the different forms of reuse and recovery of agro-residual waste is the production of biofuels. The production of second-generation ethanol has been evaluated and proposed as one of the technically viable alternatives for this purpose. This research work employed the banana pseudostem as biomass. Two different chemical pre-treatment methods (acid hydrolisis with H2SO4 2% w/w and alkaline hydrolysis with NaOH 3% w/w) of dry and milled biomass (70 g/L of dry matter, ms) were assessed, and the corresponding reducing sugars yield, AR, (YAR), after enzymatic saccharification, were determined. The effect on YAR by increasing the dry matter (ms) from 70 to 100 g/L, in dry and milled biomass and also fresh, were analyzed. Changes in cellulose crystallinity and in biomass surface morphology due to the different chemical pre-treatments were analyzed by X-ray diffraction and scanning electron microscopy. The acid pre-treatment resulted in higher YAR values, whether related to the cellulose content under saccharification (RAR = 79,48) or to the biomass concentration employed (YAR/ms = 32,8%). In a comparison between alkaline and acid pre-treatments, the latter led to an increase in the cellulose content of the reaction mixture from 52,8 to 59,8%; also, to a reduction of the cellulose crystallinity index from 51,19 to 33,34% and increases in RAR (43,1%) and YAR/ms (39,5%). The increase of dry matter (ms) bran from 70 to 100 g/L in the acid pre-treatment, resulted in a decrease of average yields in RAR (43,1%) and YAR/ms (18,2%). Using the pseudostem fresh with broth removed, whether for 70 g/L concentration or 100 g/L in dry matter (ms), similarly to the alkaline pre-treatment, has led to lower average values in RAR (67,2% and 42,2%) and in YAR/ms (28,4% e 17,8%), respectively. The acid pre-treated and saccharificated biomass broth was detoxificated with different activated carbon contents (1,2 and 4% w/v), concentrated up to AR = 100 g/L and fermented by Saccharomyces cerevisiae. The yield values (YP/AR) and productivity (QP) in ethanol were determined and compared to those values obtained from the fermentation of non-concentrated/non-detoxificated broth (AR = 18 g/L) and concentrated/non-detoxificated broth (AR = 100 g/L). The highest average value for YP/AR (0,46 g/g) was obtained from the fermentation of non-concentrated broth. This value did not present a significant difference (p<0,05) when compared to the YP/RS related to the broth concentrated and detoxificated by activated carbon 1% w/v (YP/AR = 0,41 g/g). However, a higher ethanol productivity (QP = 1,44 g/L.h) was achieved through broth detoxification. This value was 75% higher than the average QP determined using concentrated and non-detoxificated broth (QP = 0,82 g/L.h), and 22% higher than the QP found in the non-concentrated broth (QP = 1,18 g/L.h).

Keywords: biofuels, biomass, saccharification, bioethanol

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Authors: Hadiatullah, T. S. D. Desak Ketut, A. A. Ayu, A. N. Isna, D. P. Ririn

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Spirogyra sp. is genus of microalgae which has a high carbohydrate content that used as a best medium for bacterial fermentation to produce cellulose. This study objective to determine the effect of pulp banana in the fermented paper production process using Spirogyra sp. and characterizing of the paper product. The method includes the production of bacterial cellulose, assay of the effect fermented paper interpolating with banana pulp using Spirogyra sp., and the assay of paper characteristics include gram-mage paper, water assay absorption, thickness, power assay of tensile resistance, assay of tear resistance, density, and organoleptic assay. Experiments were carried out with completely randomized design with a variation of the concentration of sewage treatment in the fermented paper production interpolating banana pulp using Spirogyra sp. Each parameter data to be analyzed by Anova variance that continued by real difference test with an error rate of 5% using the SPSS. Nata production results indicate that different carbon sources (glucose and sugar) did not show any significant differences from cellulose parameters assay. Significantly different results only indicated for the control treatment. Although not significantly different from the addition of a carbon source, sugar showed higher potency to produce high cellulose. Based on characteristic assay of the fermented paper showed that the paper gram-mage indicated that the control treatment without interpolation of a carbon source and a banana pulp have better result than banana pulp interpolation. Results of control gram-mage is 260 gsm that show optimized by cardboard. While on paper gram-mage produced with the banana pulp interpolation is about 120-200 gsm that show optimized by magazine paper and art paper. Based on the density, weight, water absorption assays, and organoleptic assay of paper showing the highest results in the treatment of pulp banana interpolation with sugar source as carbon is 14.28 g/m2, 0.02 g and 0.041 g/cm2.minutes. The conclusion found that paper with nata material interpolating with sugar and banana pulp has the potential formulation to produce super-quality paper.

Keywords: cellulose, fermentation, grammage, paper, Spirogyra sp.

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Authors: Irena Maus, Katharina Gabriella Cibis, Andreas Bremges, Yvonne Stolze, Geizecler Tomazetto, Daniel Wibberg, Helmut König, Alfred Pühler, Andreas Schlüter

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Within the agricultural sector, the production of biogas from organic substrates represents an economically attractive technology to generate bioenergy. Complex consortia of microorganisms are responsible for biomass decomposition and biogas production. Recently, species belonging to the phylum Thermotogae were detected in thermophilic biogas-production plants utilizing renewable primary products for biomethanation. To analyze adaptive genome features of representative Thermotogae strains, Defluviitoga tunisiensis L3 was isolated from a rural thermophilic biogas plant (54°C) and completely sequenced on an Illumina MiSeq system. Sequencing and assembly of the D. tunisiensis L3 genome yielded a circular chromosome with a size of 2,053,097 bp and a mean GC content of 31.38%. Functional annotation of the complete genome sequence revealed that the thermophilic strain L3 encodes several genes predicted to facilitate growth of this microorganism on arabinose, galactose, maltose, mannose, fructose, raffinose, ribose, cellobiose, lactose, xylose, xylan, lactate and mannitol. Acetate, hydrogen (H2) and carbon dioxide (CO2) are supposed to be end products of the fermentation process. The latter gene products are metabolites for methanogenic archaea, the key players in the final step of the anaerobic digestion process. To determine the degree of relatedness of dominant biogas community members within selected digester systems to D. tunisiensis L3, metagenome sequences from corresponding communities were mapped on the L3 genome. These fragment recruitments revealed that metagenome reads originating from a thermophilic biogas plant covered 95% of D. tunisiensis L3 genome sequence. In conclusion, availability of the D. tunisiensis L3 genome sequence and insights into its metabolic capabilities provide the basis for biotechnological exploitation of genome features involved in thermophilic fermentation processes utilizing renewable primary products.

Keywords: genome sequence, thermophilic biogas plant, Thermotogae, Defluviitoga tunisiensis

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Authors: Ahilya Singh, Brad Carte, Ramesh Subramani, William Aalbersberg

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A total of 37 actinomycete strains were purified from 25 Solomon Islands marine sediments using four different types of isolation media. Among them, 54% of the strains had obligate requirement of seawater for growth. The ethyl acetate extract of 100 ml fermentation product of each strain was screened for antimicrobial activity against multidrug resistant human pathogens and cytotoxic activity against brine shrimps. A total of 67% of the ethyl acetate extracts showed antimicrobial and/or cytotoxic activities. A strain F-1915 was selected for isolation and evaluation of bioactive compound(s) based on its bioactive properties and chemical profile analysis using the LC-MS. The strain F-1915 was identified to have 96% sequence similarity to Streptomyces violaceusniger on the basis of 16S rDNA sequences using BLAST analysis. The 16S rDNA revealed that the strain F-1915 is a new member of MAR4 clade of actinomycetes. The MAR4 clade is an interesting clade of actinomycetes known for the production of pharmaceutically important hybrid isoprenoid compounds. The ethyl acetate extract of the fermentation product of this strain was purified by silica gel column chromatography and afforded the isolation of one bioactive pure compound. Based on the 1D and 2D NMR spectral data of compound 1 it was identified as a new mono-brominated phenazinone, Marinophenazimycin A, a structure which has already been studied by external collaborators at Scripps Institution of Oceanography but is yet to be published. Compound 1 displayed significant antimicrobial activity against drug resistant human pathogens. The minimum inhibitory concentration (MIC) of compound 1 was against Methicillin Resistant Staphylococcus aureus (MRSA) was about 1.9 μg/ml and MIC recorded against Amphotericin Resistant Candida albicans (ARCA) was about 0.24 μg/ml. The bioactivity of compound 1 against ARCA was found to be better than the standard antifungal agent amphotericin B. Compound 1 however did not show any cytotoxic activity against brine shrimps.

Keywords: actinomycetes, antimicrobial activity, brominated phenazine, MAR4 clade, marine natural products, multidrug resistent, 1D and 2D NMR

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Authors: Jong Il Rhee

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The polypeptide representing the mature part of human BMP-7 was cloned and efficiently expressed in Escherichia coli and Bacillus subtilis, which had a clear band for hBMP-7, a homodimeric protein with an apparent molecular weight of 15.4 kDa. Recombinant E.coli produced 111 pg hBMP-7/mg of protein hBMP-7 through IPTG induction. Recombinant B. subtilis also produced 350 pg hBMP-7/ml of culture medium. The hBMP-7 was purified in 2 steps using an FPLC system with an ion exchange column and a gel filtration column. The hBMP-7 produced in this work also stimulated the alkaline phosphatase (ALP) activity in a dose-dependent manner, i.e. 2.5- and 8.9-fold at 100 and 300 ng hBMP-7/ml, respectively, and showed intact biological activity.

Keywords: B. subtilis, E. coli, fermentation, hBMP-7

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Authors: Yanin Hosakun, Sujitra Wongkasemjit, Thanyalak Chaisuwan

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Carbon dioxide removal from natural gas is an important process because the existence of carbon dioxide in natural gas contributes to pipeline corrosion, reduces the heating value, and takes up volume in the pipeline. In this study, bacterial cellulose was chosen for the CO2/CH4 gas separation membrane due to its unique structure and prominent properties. Additionally, it can simply be obtained by culturing the bacteria so called “Acetobacter xylinum” through fermentation of coconut juice. Bacterial cellulose membranes with and without silver ions were prepared and studied for the separation performance of CO2 and CH4.

Keywords: bacterial cellulose, CO2, CH4 separation, membrane, nata de coco

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Authors: Camelia Vizireanu, Daniela Istrati, Alina Georgiana Profir, Rodica Mihaela Dinica

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Globally, there is a great interest in promoting the consumption of fruit and vegetables to improve health. Due to the content of essential compounds such as antioxidants, important amounts of fruits and vegetables should be included in the daily diet. Juices are good sources of vitamins and can also help increase overall fruit and vegetable consumption. Starting from this trend (introduction into the daily diet of vegetables and fruits) as well as the desire to diversify the range of functional products for both adults and children, a fermented juice was made using probiotic microorganisms based on root vegetables, with potential beneficial effects in the diet of children, vegetarians and people with lactose intolerance. The three vegetables selected for this study, red beet, carrot, and celery bring a significant contribution to functional compounds such as carotenoids, flavonoids, betalain, vitamin B and C, minerals and fiber. By fermentation, the functional value of the vegetable juice increases due to the improved stability of these compounds. The combination of probiotic microorganisms and vegetable fibers resulted in a nutrient-rich synbiotic product. The stability of the nutritional and sensory qualities of the obtained synbiotic product has been tested throughout its shelf life. The evaluation of the physico-chemical changes of the synbiotic drink during storage confirmed that: (i) vegetable juice enriched with honey and vegetable pulp is an important source of nutritional compounds, especially carbohydrates and fiber; (ii) microwave treatment used to inhibit pathogenic microflora did not significantly affect nutritional compounds in vegetable juice, vitamin C concentration remained at baseline and beta-carotene concentration increased due to increased bioavailability; (iii) fermentation has improved the nutritional quality of vegetable juice by increasing the content of B vitamins, polyphenols and flavonoids and has a good antioxidant capacity throughout the shelf life; (iv) the FTIR and Raman spectra have highlighted the results obtained using physicochemical methods. Based on the analysis of IR absorption frequencies, the most striking bands belong to the frequencies 3330 cm⁻¹, 1636 cm⁻¹ and 1050 cm⁻¹, specific for groups of compounds such as polyphenols, carbohydrates, fatty acids, and proteins. Statistical data processing revealed a good correlation between the content of flavonoids, betalain, β-carotene, ascorbic acid and polyphenols, the fermented juice having a stable antioxidant activity. Also, principal components analysis showed that there was a negative correlation between the evolution of the concentration of B vitamins and antioxidant activity. Acknowledgment: This study has been founded by the Francophone University Agency, Project Réseau régional dans le domaine de la santé, la nutrition et la sécurité alimentaire (SaIN), No. at Dunarea de Jos University of Galati 21899/ 06.09.2017 and by the Sectorial Operational Programme Human Resources Development of the Romanian Ministry of Education, Research, Youth and Sports trough the Financial Agreement POSDRU/159/1.5/S/132397 ExcelDOC.

Keywords: bioactive compounds, fermentation, synbiotic drink from vegetables, stability during storage

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Authors: Rouzbeh Jafari, Joe Nava

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This study includes learnings from an engineering practice normally performed on large scale biohydrogen processes. If properly scale-up is done, biohydrogen can be a reliable pathway for biowaste valorization. Most of the studies on biohydrogen process development have used model feedstock to investigate process key performance indicators (KPIs). This study does not intend to compare different technologies with model feedstock. However, it reports economic drivers and technical challenges which help in developing a road map for expanding biohydrogen economy deployment in Canada. BBA is a consulting firm responsible for the design of hydrogen production projects. Through executing these projects, activity has been performed to identify, register and mitigate technical drawbacks of large-scale hydrogen production. Those learnings, in this study, have been applied to the biohydrogen process. Through data collected by a comprehensive literature review, a base case has been considered as a reference, and several case studies have been performed. Critical parameters of the process were identified and through common engineering practice (process design, simulation, cost estimate, and life cycle assessment) impact of these parameters on the commercialization risk matrix and class 5 cost estimations were reported. The process considered in this study is food waste and woody biomass dark fermentation. To propose a reliable road map to develop a sustainable biohydrogen production process impact of critical parameters was studied on the end-to-end process. These parameters were 1) feedstock composition, 2) feedstock pre-treatment, 3) unit operation selection, and 4) multi-product concept. A couple of emerging technologies also were assessed such as photo-fermentation, integrated dark fermentation, and using ultrasound and microwave to break-down feedstock`s complex matrix and increase overall hydrogen yield. To properly report the impact of each parameter KPIs were identified as 1) Hydrogen yield, 2) energy consumption, 3) secondary waste generated, 4) CO2 footprint, 5) Product profile, 6) $/kg-H2 and 5) environmental impact. The feedstock is the main parameter defining the economic viability of biohydrogen production. Through parametric studies, it was found that biohydrogen production favors feedstock with higher carbohydrates. The feedstock composition was varied, by increasing one critical element (such as carbohydrate) and monitoring KPIs evolution. Different cases were studied with diverse feedstock, such as energy crops, wastewater slug, and lignocellulosic waste. The base case process was applied to have reference KPIs values and modifications such as pretreatment and feedstock mix-and-match were implemented to investigate KPIs changes. The complexity of the feedstock is the main bottleneck in the successful commercial deployment of the biohydrogen process as a reliable pathway for waste valorization. Hydrogen yield, reaction kinetics, and performance of key unit operations highly impacted as feedstock composition fluctuates during the lifetime of the process or from one case to another. In this case, concept of multi-product becomes more reliable. In this concept, the process is not designed to produce only one target product such as biohydrogen but will have two or multiple products (biohydrogen and biomethane or biochemicals). This new approach is being investigated by the BBA team and the results will be shared in another scientific contribution.

Keywords: biohydrogen, process scale-up, economic evaluation, commercialization uncertainties, hydrogen economy

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Authors: Hamidreza Khodaei, Ali Daryabeigi Zand

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Conjugated linoleic acid (CLA) is a mixture of isomers of linoleic acid. Despite the fact that 28 different isomers of CLA have already been identified, but the main isomer found in natural diets more than ninety percent CLA on intake of food constitutes demonstrates. CLA is known to be a substance that readily available by rumen microorganisms in some ruminants such as cattle and sheep would likely be made. The main objective of this research was to evaluate the impacts of CLA on lipid metabolism and enhanced fat around the muscle durability by reducing the process of oxidation. In order to implement this research, 80 female mice of the Balb/C, with 55 days of age were employed in the experiment. Treatments include various levels of CLA. Over the course of this study blood samples was also taken from the tail vein of the studied mice. Some other relevant parameters such as serum concentrations of triglycerides, total cholesterol, LDL, HDL and liver enzymes were also determined. The oxidative stability of fats TBARS technique was investigated at different intervals. The findings of the research were analyzed by statistical software of SAS 98. The results, CLA had no significant effect on liver enzymes (P > 0.05). However, it showed a statistically significant impact on triglycerides and total cholesterol. Ratio of LDL to HDL declined remarkably. Histological studies demonstrated reduced accumulation of fat in the tissues surrounding muscles.

Keywords: conjugated linoleic acid, fat metabolism, fat retention, oxidation process

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Authors: Nina Tsvetkova, Rumen Harizanov, Aleksandra Ivanova, Iskra Rainova, Nina Yancheva-Petrova, Dimitar Strashimirov, Raina Enikova, Mihaela Videnova, Eleonora Kaneva, Iskren Kaftandjiev, Viktoria Levterova, Ivan Simeonovski, Nikolay Yanev, Georgi Hinkov

Abstract:

The Pneumocystis jirovecii (P. jirovecii) and protozoan of the genera Acanthamoeba, Cryptosporidium, and Toxoplasma gondii are opportunistic pathogens that can cause life-threatening infections in immunocompromised patients. Aim of the study was to evaluate the coinfection rate with opportunistic protozoal agents among Bulgarian patients diagnosed with P. jirovecii pneumonia. Thirty-eight pulmonary samples were collected from 38 patients (28 HIV-infected) with P. jirovecii infection. P. jirovecii DNA was detected by real-time PCR targeting the large mitochondrial subunit ribosomal RNA gene. Acanthamoeba was determined by genus-specific conventional PCR assay. Real-time PCR for the detection of a Toxoplasma gondii and Cryptosporidium DNA fragment was used. Pneumocystis DNA was detected in all 38 specimens; 28 (73.7%) were from HIV-infected patients. Three (10,7%) of them were co-infected with T. gondii and 1 (3.6%) with Cryptosporidium. In the group of non-HIV-infected (n=10), Cryptosporidium DNA was detected in an infant (10%). Acanthamoeba DNA was not found in the tested samples. The current study showed a relatively low rate of coinfections of Cryptosporidium spp./T. gondii and P. jirovecii in the Bulgarian patients studied.

Keywords: coinfection, opportunistic protozoal agents, Pneumocystis jirovecii, pulmonary infections

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Authors: Olfat A. Mohamed

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Rhamnolipids biosurfactants have distinct properties given them importance in many industrial applications, especially their great new future applications in cosmetic and pharmaceutical industries. These applications have encouraged the search for diverse and renewable resources to control the cost of production. The experimental results were then applied to find a suitable mathematical model for obtaining the design criteria of the batch bioreactor. This research aims to produce Rhamnolipids from different oily wastewater sources such as petroleum crude oil (PO) and vegetable oil (VO) by using Pseudomonas aeruginosa ATCC 9027. Different concentrations of the PO and the VO are added to the media broth separately are in arrangement (0.5 1, 1.5, 2, 2.5 % v/v) and (2, 4, 6, 8 and 10%v/v). The effect of the initial concentration of oil residues and the addition of glycerol and palmitic acid was investigated as an inducer in the production of rhamnolipid and the surface tension of the broth. It was found that 2% of the waste (PO) and 6% of the waste (VO) was the best initial substrate concentration for the production of rhamnolipids (2.71, 5.01 g rhamnolipid/l) as arrangement. Addition of glycerol (10-20% v glycerol/v PO) to the 2% PO fermentation broth led to increase the rhamnolipid production (about 1.8-2 times fold). However, the addition of palmitic acid (5 and 10 g/l) to fermentation broth contained 6% VO rarely enhanced the production rate. The experimental data for 2% initially (PO) was used to estimate the various kinetic parameters. The following results were obtained, maximum rate or velocity of reaction (Vmax) = 0.06417 g/l.hr), yield of cell weight per unit weight of substrate utilized (Yx/s = 0.324 g Cx/g Cs) maximum specific growth rate (μmax = 0.05791 hr⁻¹), yield of rhamnolipid weight per unit weight of substrate utilized (Yp/s)=0.2571gCp/g Cs), maintenance coefficient (Ms =0.002419), Michaelis-Menten constant, (Km=6.1237 gmol/l), endogenous decay coefficient (Kd=0.002375 hr⁻¹). Predictive parameters and advanced mathematical models were applied to evaluate the time of the batch bioreactor. The results were as follows: 123.37, 129 and 139.3 hours in respect of microbial biomass, substrate and product concentration, respectively compared with experimental batch time of 120 hours in all cases. The expected mathematical models are compatible with the laboratory results and can, therefore, be considered as tools for expressing the actual system.

Keywords: batch bioreactor design, glycerol, kinetic parameters, petroleum crude oil, Pseudomonas aeruginosa, rhamnolipids biosurfactants, vegetable oil

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Authors: Michael Graz, Andrew Shearer, Gareth Evans

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Manipulation of rumen bacteria is receiving increasing attention as a way of controlling greenhouse gas (GHG) emissions that are generated by the agricultural sector. Feed supplementation in particular is one of the ways in which this drive is being addressed, in particular with reference to livestock-generated GHG emissions. A blend of naturally occurring chemical extracts obtained from garlic and bitter orange extracts has been identified as a natural, sustainable and non-antibiotic based way of reducing methane production by ruminant livestock. In the current study, the acceptability and impact of this blend of natural extracts on feed rations of beef cattle was trialed in vivo on a commercial farm in Europe. Initial findings have demonstrated acceptable palatability, with all animals accepting the feed supplement into their ration both when it was mixed into the total daily ration and when used as a part of their high energy rations. Measurement of the impact of this feed supplement on productivity weight gain and milk quality is ongoing. In conclusion, this field study confirmed the palatability of the combination of garlic and citrus extracts and hence pointed to possibility of the extract blend to improve digestion, enhance body energy retention and limit CH4 formation in relation to feed intake.

Keywords: citrus, garlic, methane reduction, palatability, ruminants

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Authors: Thabiso M. Sebolai, Victor Mlambo, Solomon Tefera, Othusitse R. Madibela

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In some tree species, sustained herbivory can induce changes in biosynthetic pathways resulting in overproduction of anti-nutritional secondary plant compounds. This inductive mechanism, which has not been demonstrated in semi-arid rangelands of South Africa, may result in browse leaves of lower nutritive value. In this study we investigate the interactive effect of browsing pressure and tree species on chemical composition, buffer nitrogen solubility index (NSI), in vitro ruminal dry matter degradability (IVDMD) and in vitro ruminal N degradability (IVND) of leaves. Leaves from Maytenus capitata, Olea africana, Coddia rudis, Carissa macrocarpa, Rhus refracta, Ziziphus mucronata, Boscia oliedes, Grewia robusta, Phyllanthus vessucosus and Ehretia rigida trees growing in a communal grazing area were harvested at two heights: browsable ( < 1.5 m) and non-browsable ( > 1.5 m), representing high and low browsing pressure, respectively. The type of animals utilizing the communal rangeland includes cattle at 1 livestock unit (450kg)/12 to 15 hectors and goats at 1 livestock unit/4 ha. Harvested leaves were dried, milled and analysed for proximate components, soluble phenolics, condensed tannins, minerals and in vitro ruminal fermentation. A significant plant species and harvesting height interaction effect (P < 0.05) was observed for total nitrogen (N) and soluble phenolics concentration. Tree species and harvesting height affected (P < 0.05) condensed tannin (CTs) content where samples harvested from the non-browsable height had higher (0.61 AU550 nm/200 mg) levels than those harvested at browsable height (0.55 AU550 nm/200 mg) while their interaction had no effects. Macro and micro-minerals were only influenced (P < 0.05) by browse species but not harvesting height. Species and harvesting height interacted (P < 0.05) to influence IVDMD and IVND of leaves at 12, 24 and 36 hours of incubation. The different browse leaves contained moderate to high protein, moderate level of phenolics and minerals, suggesting that they have the potential to provide supplementary nutrients for ruminants during the dry seasons.

Keywords: browse plants, chemical composition, harvesting heights, phenolics

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Authors: Peter C. St. John, Michael F. Crowley, Yannick J. Bomble

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Production of chemicals from engineered organisms in a batch culture typically involves a trade-off between productivity, yield, and titer. However, strategies for strain design typically involve designing mutations to achieve the highest yield possible while maintaining growth viability. Such approaches tend to follow the principle of designing static networks with minimum metabolic functionality to achieve desired yields. While these methods are computationally tractable, optimum productivity is likely achieved by a dynamic strategy, in which intracellular fluxes change their distribution over time. One can use multi-stage fermentations to increase either productivity or yield. Such strategies would range from simple manipulations (aerobic growth phase, anaerobic production phase), to more complex genetic toggle switches. Additionally, some computational methods can also be developed to aid in optimizing two-stage fermentation systems. One can assume an initial control strategy (i.e., a single reaction target) in maximizing productivity - but it is unclear how close this productivity would come to a global optimum. The calculation of maximum theoretical yield in metabolic engineering can help guide strain and pathway selection for static strain design efforts. Here, we present a method for the calculation of a maximum theoretical productivity of a batch culture system. This method follows the traditional assumptions of dynamic flux balance analysis: that internal metabolite fluxes are governed by a pseudo-steady state and external metabolite fluxes are represented by dynamic system including Michealis-Menten or hill-type regulation. The productivity optimization is achieved via dynamic programming, and accounts explicitly for an arbitrary number of fermentation stages and flux variable changes. We have applied our method to succinate production in two common microbial hosts: E. coli and A. succinogenes. The method can be further extended to calculate the complete productivity versus yield Pareto surface. Our results demonstrate that nearly optimal yields and productivities can indeed be achieved with only two discrete flux stages.

Keywords: A. succinogenes, E. coli, metabolic engineering, metabolite fluxes, multi-stage fermentations, succinate

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Authors: Şevket Evci, Mehmet Akif Karsli

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The aim of this study is to determine nutrient contents, in situ ruminal degradation kinetics and protein fractions of screenings bean (B), chick pea (ChP), red lentil (RL) and green lentil (GL) that is used as residue in grain legume packing industry. For this purpose, four samples of each legumes species-a total of 16 samples, collected from different parts of our country were utilized. Feedstuffs used in the experiment were incubated for 0, 2 4, 8, 12, 24, and 48 hours in the rumen of 3 ruminally cannulated Akkaraman rams as duplicate. The nutrient contents, in situ ruminal dry matter (DM), organic matter (OM) and crude protein (CP) degradabilities and fractions, and escape protein contents were evaluated. The highest OM and CP contents were observed in RL (P<0.05). Chick pea had the highest ether extract (EE) content and EE values were 3.47, 6.72, 2.26, 8.66 % for RL, B, GL and ChP, respectively (P<0.05). Crude fiber (CF), ADF, and NDF contents were the highest in RL and the lowest in ChP. CF values were 24.03, 10.80, 4.09 and 3.57 % for RL, GL, B and ChP (P<0.05). Acid detergent insoluble nitrogen content of samples did not differ. Escape protein content was the highest in RL and the lowest in B (P<0.05). After 48 h incubation, the lowest OM and CP degradabilities were observed in RL. While the highest OM degradability was seen in ChP the highest CP degradability was observed in B (P<0.05). The lowest water soluble OM and CP contents were observed in RL whereas the highest potentially degradable OM and CP contents were seen in B and ChP (P<0.05). Both rate of OM and CP degradations (k-1) did not differ among samples (P>0.05). In conclusion, it was noted that feedstuffs (GL, ChP and B) used in the experiment except RL had a greater ruminal degradibilities of both OM and CP and moreover, had a higher escape protein contents, except B. It was thought that these feedstuffs can be substituted with some of common protein sources used in animal nutrition.

Keywords: in situ, nutrient contents, ruminant, subsieve

Procedia PDF Downloads 450