Search results for: ruminal fermentation

Authors: Shady S. Hassan, Brijesh K. Tiwari, Gwilym A. Williams, Amit K. Jaiswal

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EU is known as the destination with the highest rate of the coffee consumption per capita in the world. Spent coffee grounds (SCG) are the main by-product of coffee brewing. SCG is either disposed as a solid waste or employed as compost, although the polysaccharides from such lignocellulosic biomass might be used as feedstock for fermentation processes. However, SCG as a lignocellulose have a complex structure and pretreatment process is required to facilitate an efficient enzymatic hydrolysis of carbohydrates. However, commonly used pretreatment methods, such as chemical, physico-chemical and biological techniques are still insufficient to meet optimal industrial production requirements in a sustainable way. Ultrasound is a promising candidate as a sustainable green pretreatment solution for lignocellulosic biomass utilization in a large scale biorefinery. Thus, ultrasound pretreatment of SCG without adding harsh chemicals investigated as a green technology to enhance enzyme hydrolysis. In the present work, ultrasound pretreatment experiments were conducted on SCG using different ultrasound frequencies (25, 35, 45, 130, and 950 kHz) for 60 min. Regardless of ultrasound power, low ultrasound frequency is more effective than high ultrasound frequency in pretreatment of biomass. Ultrasound pretreatment of SCG (at ultrasound frequency of 25 kHz for 60 min) followed by enzymatic hydrolysis resulted in total reducing sugars of 56.1 ± 2.8 mg/g of biomass. Fourier transform Infrared Spectroscopy (FTIR) was employed to investigate changes in functional groups of biomass after pretreatment, while high-performance liquid chromatography (HPLC) was employed for determination of glucose. Pretreatment of lignocellulose by low frequency ultrasound in water only was found to be an effective green approach for SCG to improve saccharification and glucose yield compared to native biomass. Pretreatment conditions will be optimized, and the enzyme hydrolysate will be used as media component substitute for the production of ethanol.

Keywords: lignocellulose, ultrasound, pretreatment, spent coffee grounds

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Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

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The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever-growing accepted option in the way to the development of biorefinery complexes; in the Mexican state of Sinaloa, two million tons of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second-generation bioethanol (2G), enzymes, and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our workgroup. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU / mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol

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Authors: Mauricio Cruz, Andrés Díaz García, Martha Isabel Gómez, Juan Carlos Serrato Bermúdez

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The disadvantages of using natural substrates in SSF processes have been well recognized and mainly are associated to gradual decomposition of the substrate, formation of agglomerates and decrease of porosity bed generating limitations in the mass and heat transfer. Additionally, in several cases, materials with a high agricultural value such as sour milk, beets, rice, beans and corn have been used. Thus, the use of economic inert supports (natural or synthetic) in combination with a nutrient suspension for the production of biocontrol microorganisms is a good alternative in SSF processes, but requires further studies in the fields of modeling and optimization. Therefore, the aim of this work is to compare the performance of two inert supports, a synthetic (polyurethane foam) and a natural one (rice husk), identifying the factors that have the major effects on the productivity of T. asperellum Th204 and the maximum specific growth rate in a PROPHYTA L05® fixed bed bioreactor. For this, the six factors C:N ratio, temperature, inoculation rate, bed height, air moisture content and airflow were evaluated using a fractional design. The factors C:N and air flow were identified as significant on the productivity (expressed as conidia/dry substrate•h). The polyurethane foam showed higher maximum specific growth rate (0.1631 h-1) and productivities of 3.89 x107 conidia/dry substrate•h compared to rice husk (2.83x106) and natural substrate based on rice (8.87x106) used as control. Finally, a quadratic model was generated and validated, obtaining productivities higher than 3.0x107 conidia/dry substrate•h with air flow at 0.9 m3/h and C:N ratio at 18.1.

Keywords: bioprocess, scale up, fractional design, C:N ratio, air flow

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Authors: Miguel Mellado, Cecilia Zapata

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Goats can be exploited in harsh environments due to their capacity to adjust to limited quantity and quality forage sources. In these environments, leguminous trees can be used as supplementary feeds as foliage and fruits of these trees can contribute to maintain or improve production efficiency in ruminants. The objective of this study was to determine the nutritional value of three leguminous trees heavily selected by goats in a xerophyllous shrubland. Chemical composition and in vitro dry matter disappearance (IVDMD) of leaves and pods from leucaena (Leucaena leucocephala), mesquite (Prosopis laevigata) and huisache (Acacia farnesiana) is presented. Crude protein (CP) ranged from 17.3% for leaves of huisache to 21.9% for leucaena. The neutral detergent fiber (NDF) content ranged from 39.0 to 40.3 with no difference among fodder threes. Across tree species, mean IVDMD was 61.6% for pods and 52.2% for leaves. IVDMD for leaves was highest (P < 0.01) for leucaena (54.9%) and lowest for huisache (47.3%). Condensed tannins in an acetonic extract were highest for leaves of huisache (45.3 mg CE/g DM) and lowest for mesquite (25.9 mg CE/g DM). Pods and leaves of huisache presented the highest number of secondary metabolites, mainly related to hydrobenzoic acid and flavonols; leucaena and mesquite presented mainly flavonols and anthocyanins. It was concluded that leaves and pods of leucaena, mesquite and huisache constitute valuable forages for ruminant livestock due to its low fiber, high CP levels, moderate in vitro fermentation characteristics and high mineral content. Keywords: Fodder tree; ruminants; secondary metabolites; minerals; tannins

Keywords: fodder tree, ruminants, secondary metabolites, minerals, tannins

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Authors: Gafar Bamigbade, Adebunkola Omemu

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“Ogi” is a fermented cereal gruel prepared from maize (Zea mays), millet (Pennisetum typhoideum) or guinea corn (Sorghum bicolour). It could be boiled to give a thicker consistency wrapped in leaf allowed to cool and set to a gel known as “eko”. The objective of this study is to determine the amylase activities of mould associated with the spoilage of Ogi and eko. Moulds were isolated from spoilt Ogi and eko samples using standard microbiological procedures. The isolate was then screened for amylase production using starch agar medium. Positive isolates were used for amylase production by solid state fermentation (SFF) using rice bran as the medium. An alpha-amylase and glucoamylase activity of the crude enzyme was determined using the DNS method. The mean mold Population ranged from 1.15 X 105cfu/g for raw Ogi to 6.25 X 105cfu/g for Eko (wrapped in Leaves). Twenty-seven (27) moulds isolated from the sample include A. niger, A. flavus, A. fumigatus, Rhizopus species and Penicillium species. Aspergillus flavus had the highest percentage (51.9%) of incidence while Penicillium species had the least (3.7%). Out of the 27 isolates screened, 19 were found to be amylase positive by showing a clear zone around their colony after flooding with iodine solution. Diameter of clear zone ranged from 3.00mm (Aspergillus niger, C4) to 22.00mm (Aspergillus flavus, A1). Aspergillus niger isolated from spoilt Eko wrapped in leaf has the highest percentage alpha-amylase activity (30.8%) and Aspergillus flavus isolated from spoilt raw ogi has the lowest activity (11.4%). Aspergillus niger isolated from spoilt Eko wrapped in nylon produces the highest glucoamylase activity (240U/ml) while penicillium specie isolated from spoilt cooked ogi has the lowest activity (100U/ml). This study shows that moulds associated with spoilage of ogi and eko can produce amylase.

Keywords: glucoamylase, alpha amylase, ogi, eko

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Authors: Debajyoti Bose

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Chitosan has received much attention as a functional biopolymer for diverse applications, especially in pharmaceutics and medicine. Chitosan is a positively charged natural biodegradable and biocompatible polymer. It is a linear polysaccharide consisting of β-1,4 linked monomers of glucosamine and N-acetylglucosamine. Chitosan can be mainly obtained from fungal sources during large fermentation process. In this study,three different fungal strains Aspergillus niger NCIM 1045, Aspergillus oryzae NCIM 645 and Mucor indicus MTCC 3318 were used for the production of chitosan. The growth mediums were optimized for maximum fungal production. The produced chitosan was characterized by determining degree of deacetylation. Chitosan possesses one reactive amino at the C-2 position of the glucosamine residue, and these amines confer important functional properties to chitosan which can be exploited for biofabrication to generate various chemically modified derivatives and explore their potential for pharmaceutical field. Chitosan nanoparticles were prepared by ionic cross-linking with tripolyphosphate (TPP). The major effect on encapsulation and release of protein (e.g. enzyme diastase) in chitosan-TPP nanoparticles was investigated in order to control the loading and release efficiency. It was noted that the chitosan loading and releasing efficiency as a nanocapsule, obtained from different fungal sources was almost near to initial enzyme activity(12026 U/ml) with a negligible loss. This signify, chitosan can be used as a polymeric drug as well as active component or protein carrier material in dosage by design due to its appealing properties such as biocompatibility, biodegradability, low toxicity and relatively low production cost from abundant natural sources. Based upon these initial experiments, studies were also carried out on modification of chitosan based nanocapsules incorporated with physiologically important enzymes and nutraceuticals for target delivery.

Keywords: fungi, chitosan, enzyme, nanocapsule

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Authors: Eng Toon Saw, Kun Liang Ang, Wei He, Xuecheng Dong, Seeram Ramakrishna

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Solvent recovery process is receiving utmost attention in recent year due to the scarcity of natural resource and consciousness of circular economy in chemical and pharmaceutical manufacturing process. Solvent dehydration process is one of the important process to recover and to purify the solvent for reuse. Due to the complexity of solvent waste or wastewater effluent produced in pharmaceutical industry resulting the wastewater treatment process become complicated, thus an alternative solution is to recover the valuable solvent in solvent waste. To treat solvent waste and to upgrade solvent purity, membrane pervaporation process is shown to be a promising technology due to the energy intensive and low footprint advantages. Ceramic membrane is adopted as solvent dehydration membrane owing to the chemical and thermal stability properties as compared to polymeric membrane. NaA zeolite membrane is generally used as solvent dehydration process because of its narrow and distinct pore size and high hydrophilicity. NaA zeolite membrane has been mainly applied in alcohol dehydration in fermentation process. At this stage, the membrane performance exhibits high separation factor with low flux using tubular ceramic membrane. Thus, defect free and ultrathin NaA membrane should be developed to increase water flux. Herein, we report a simple preparation protocol to prepare ultrathin NaA zeolite membrane supported on tubular ceramic membrane by controlling the seed size synthesis, seeding methods and conditions, ceramic substrate surface pore size selection and secondary growth conditions. The microstructure and morphology of NaA zeolite membrane will be examined and reported. Moreover, the membrane separation performance and stability will also be reported in isopropanol dehydration, ketone dehydration and ester dehydration particularly for the application in pharmaceutical industry.

Keywords: ceramic membrane, NaA zeolite, pharmaceutical industry, solvent recovery

Procedia PDF Downloads 245

Authors: Hadjer Sadoune, Liza Lamini, Scherazade Krim, Amel Djouadi, Rachida Rihani

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Deep neural networks are highly regarded for their accuracy in predicting intricate fermentation processes. Their ability to learn from a large amount of datasets through artificial intelligence makes them particularly effective models. The primary obstacle in improving the performance of these models is to carefully choose the suitable hyperparameters, including the neural network architecture (number of hidden layers and hidden units), activation function, optimizer, learning rate, and other relevant factors. This study predicts biogas production from real wastewater treatment plant data using a sophisticated approach: hybrid Bayesian optimization with a tree-structured Parzen estimator (BO-TPE) for an optimised deep neural network (DNN) model. The plant utilizes an Upflow Anaerobic Sludge Blanket (UASB) digester that treats industrial wastewater from soft drinks and breweries. The digester has a working volume of 1574 m3 and a total volume of 1914 m3. Its internal diameter and height were 19 and 7.14 m, respectively. The data preprocessing was conducted with meticulous attention to preserving data quality while avoiding data reduction. Three normalization techniques were applied to the pre-processed data (MinMaxScaler, RobustScaler and StandardScaler) and compared with the Non-Normalized data. The RobustScaler approach has strong predictive ability for estimating the volume of biogas produced. The highest predicted biogas volume was 2236.105 Nm³/d, with coefficient of determination (R2), mean absolute error (MAE), and root mean square error (RMSE) values of 0.712, 164.610, and 223.429, respectively.

Keywords: anaerobic digestion, biogas production, deep neural network, hybrid bo-tpe, hyperparameters tuning

Procedia PDF Downloads 38

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

Procedia PDF Downloads 321

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

Procedia PDF Downloads 80

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

Procedia PDF Downloads 369

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

Procedia PDF Downloads 367

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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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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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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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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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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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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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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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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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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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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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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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™

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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

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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

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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

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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

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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

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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