Search results for: yeast fermentation

Authors: Yahia Massinissa, Afaf Benhouda, Mouloud Yahia

Abstract:

Objective: The aim of this study was to investigate the toxicity; analgesic and anti-pyretic properties of standardized HA methanolic extract (HAMeOH) in vivo. Methods: The acute toxicity study was performed on rats while adopting the OECD-420 Guidelines (fixed dose procedure). Assessment of analgesic activity was performed in rats with two analgesic models. One was acetic acid induced writhing response and the other formalin-induced paw licking. The anti-pyretic effect was tested by Brewer’s yeast induced fever in rats. Results: For the acute toxicity test, the higher dose administration of 2000 mg/kg bw. of H.albus did not produce any toxic signs or deaths in rats. There were no significant differences (p>0.05) in the body and organ weights between control and treated groups. The (LD50) of 'H. albus' was higher than 2000 g/kg bw. In subacute toxicity study, no mortality and toxic signs were observed with the doses of 100 and 200 mg/kg bw. of extracts of for 28 consecutive days. These analgesic experimental results indicated that HAMeOH (100 mg/kg and 200 mg/kg) decreased the acetic acid-induced writhing responses and HAMeOH (100 mg/kg and 200 mg/kg) decreased the licking time in the second phase of the formalin test. Moreover, in the model of yeast-induced elevation of the body temperature HAMeOH showed dose-dependent lowering of the body temperature up to 3h at both the doses these results obtained, were comparable to that of paracetamol. Conclusion: The present findings indicate that the leaves of Hyoscyamus albus L. possess potent analgesic and antipyretic activity.

Keywords: Hyoscyamus albus, Umbilicus rupestris, secondary metabolites, NMR with protons, pharmacobiologic activities, methanolic extract

Procedia PDF Downloads 400

Authors: Zakaria Boual, Abdellah Kemassi, Toufik Chouana, Philippe Michaud, Mohammed Didi Ould El Hadj

Abstract:

In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% are neutral sugar and the rest 16.25±1.62% are uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructo-oligosaccharide (RP95). The oligosaccharides concentration was 1g/L of man rogosa sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Keywords: Zizyphus lotus, polysaccharides, characterization, prebiotic effects

Procedia PDF Downloads 389

Authors: András Fülöp, Lejla Heilmann, Zsolt Szabó, Ákos Koós

Abstract:

Microbial fuel cells (MFCs) present a sustainable biotechnological solution to future energy demands. The aim of this study was to construct soil based, single cell, membrane-less MFC systems, operated without treatment to continuously power on-site monitoring and control systems during the soil bioremediation processes. Our Pseudomonas aeruginosa 541 isolate is an ideal choice for MFCs, because it is able to produce pyocyanin which behaves as electron-shuttle molecule, furthermore, it also has a significant antimicrobial effect. We tested several materials and structural configurations to obtain long term high power output. Comparing different configurations, a proton exchange membrane-less, 0.6 m long with 0.05 m diameter MFC tubes offered the best long-term performances. The long-term electricity production were tested from starch, yeast extract (YE), carboxymethyl cellulose (CMC) with humic acid (HA) as a mediator. In all cases, 3 kΩ external load have been used. The two best-operated systems were the Pseudomonas aeruginosa 541 containing MFCs with 1 % carboxymethyl cellulose and the MFCs with 1% yeast extract in the anode area and 35% hydrogel in the cathode chamber. The first had 3.3 ± 0.033 mW/m2 and the second had 4.1 ± 0.065 mW/m2 power density values. These systems have operated for 230 days without any treatment. The addition of 0.2 % HA and 1 % YE referred to the volume of the anode area resulted in 1.4 ± 0.035 mW/m2 power densities. The mixture of 1% starch with 0.2 % HA gave 1.82 ± 0.031 mW/m2. Using CMC as retard carbon source takes effect in the long-term bacterial survivor, thus enable the expression of the long term power output. The application of hydrogels in the cathode chamber significantly increased the performance of the MFC units due to their good water retention capacity.

Keywords: microbial fuel cell, bioremediation, Pseudomonas aeruginosa, biotechnological solution

Procedia PDF Downloads 272

Authors: Temitope D. Awobusuyi, Eric O. Amonsou, Muthulisi Siwela

Abstract:

Amahewu, a lactic acid fermented non-alcoholic maize based beverage is widely consumed in Southern Africa. It is traditionally made with white maize which is deficient in vitamin A. Provitamin A-biofortified maize has been developed for use as a complementary strategy to alleviate vitamin A deficiency. In this study the nutritional composition and protein digestibility of amahewu produced using provitamin A-biofortified maize was determined. Provitamin A-biofortified amahewu was processed by fermenting cooked maize porridge using malted provitamin A-biofortified maize, wheat bran and lactobacillus mixed starter culture with either malted maize or wheat bran. The total provitamin A content in amahewu products ranged from 3.3-3.8 μg/g (DW). The % retention of total provitamin A ranged from 79 %- 90 % μg/g (DW). The lowest % retention was observed in products fermented with the addition of starter culture. The gross energy of amahewu products were approx. 20 MJ/kg. There was a slight increase in the lysine content of amahewu after fermentation. Protein digestibility of amahewu (approx.91%) was slightly higher compared to unprocessed provitamin A maize (86%). However, a general decrease was observed in the minerals when compared to the unprocessed provitamin A maize. Amahewu processed using starter cultures has higher iron content than those processed with the addition of malt. These result suggests that provitamin A-biofortified amahewu has the potential to make a significant contribution towards alleviating Vitamin A Deficiency in rural communities who are also the most vulnerable to VAD.

Keywords: vitamin A deficiency, provitamin A maize, biofortification, fermentation

Procedia PDF Downloads 394

Authors: Chukwuma S. Ezeonu, Ikechukwu N. E. Onwurah, Uchechukwu U. Nwodo, Chibuike S. Ubani, Chigozie M. Ejikeme

Abstract:

Trichophyton soudanense and Trichophyton mentagrophyte were isolated from the rice mill environment, cultured and used singly and as di-culture in the treatment of measure quantities of preheated rice husk. Optimized conditions studied showed that carboxymethylcellulase (CMCellulase) activity of 57.61 µg/ml/min was optimum for Trichophyton mentagrophyte heat pretreated rice husk crude enzymes at 50oC and 80oC respectively. Duration of 120 hours (5 days) gave the highest CMcellulase activity of 75.84 µg/ml/min for crude enzyme of Trichophyton mentagrophyte heat pretreated rice husk. However, 96 hours (4 days) duration gave maximum activity of 58.21 µg/ml/min for crude enzyme of Trichophyton soudanense heat pretreated rice husk. Highest CMCellulase activities of 67.02 µg/ml/min and 69.02 µg/ml/min at pH of 5 were recorded for crude enzymes of monocultures of Trichophyton soudanense (TS) and Trichophyton mentagrophyte (TM) heat pretreated rice husk respectively. Biomass components showed that rice husk cooled after heating followed by treatment with Trichophyton mentagrophyte gave 44.50 ± 10.90 (% ± Standard Error of Mean) cellulose as the highest yield. Maximum total lignin value of 28.90 ± 1.80 (% ± SEM) was obtained from pre-heated rice husk treated with di-culture of Trichophyton soudanense and Trichophyton mentagrophyte (TS+TM). The hemicellulose content of 30.50 ± 2.12 (% ± SEM) from pre-heated rice husk treated with Trichophyton soudanense (TS); lignin value of 28.90 ± 1.80 from pre-heated rice husk treated with di-culture of Trichophyton soudanense and Trichophyton mentagrophyte (TS+TM); also carbohydrate content of 16.79 ± 9.14 (% ± SEM) , reducing and non-reducing sugar values of 2.66 ± 0.45 and 14.13 ± 8.69 (% ± SEM) were all obtained from for pre- heated rice husk treated with Trichophyton mentagrophyte (TM). All the values listed above were the highest values obtained from each rice husk treatment. The pre-heated rice husk treated with Trichophyton mentagrophyte (TM) fermented with palmwine yeast gave bio-ethanol value of 11.11 ± 0.21 (% ± Standard Deviation) as the highest yield.

Keywords: Trichophyton soudanense, Trichophyton mentagrophyte, biomass, bioethanol, rice husk

Procedia PDF Downloads 657

Authors: M. Mischopoulou, P. Naidis, S. Kalamaras, T. Kotsopoulos, P. Samaras

Abstract:

The treatment of high strength wastewater by an Upflow Anaerobic Sludge Blanket (UASB) reactor has several benefits, such as high organic removal efficiency, short hydraulic retention time along with low operating costs. In addition, high volumes of biogas are released in these reactors, which can be utilized in several industrial facilities for energy production. This study aims at the examination of the application potential of anaerobic treatment of wastewater, with high molasses content derived from yeast manufacturing, by a lab-scale UASB reactor. The molasses wastewater and the sludge used in the experiments were collected from the wastewater treatment plant of a baker’s yeast manufacturing company. The experimental set-up consisted of a 15 L thermostated UASB reactor at 37 ◦C. Before the reactor start-up, the reactor was filled with sludge and molasses wastewater at a ratio 1:1 v/v. Influent was fed to the reactor at a flowrate of 12 L/d, corresponding to a hydraulic residence time of about 30 h. Effluents were collected from the system outlet and were analyzed for the determination of the following parameters: COD, pH, total solids, volatile solids, ammonium, phosphates and total nitrogen according to the standard methods of analysis. In addition, volatile fatty acid (VFA) composition of the effluent was determined by a gas chromatograph equipped with a flame ionization detector (FID), as an indicator to evaluate the process efficiency. The volume of biogas generated in the reactor was daily measured by the water displacement method, while gas composition was analyzed by a gas chromatograph equipped with a thermal conductivity detector (TCD). The effluent quality was greatly enhanced due to the use of the UASB reactor and high rate of biogas production was observed. The anaerobic treatment of the molasses wastewater by the UASB reactor improved the biodegradation potential of the influent, resulting at high methane yields and an effluent with better quality than the raw wastewater.

Keywords: anaerobic digestion, biogas production, molasses wastewater, UASB reactor

Procedia PDF Downloads 255

Authors: Saranya Ganapathy, Megha N. Parajulee, Michael San Francisco, Hong Zhang

Abstract:

Insect pests are a serious threat to agricultural productivity. Use of chemical pesticides, the predominant control method thus far, has resulted in environmental damage, pest resurgence, and negative effects on non-target species. Genetically modified (GM) crops offer a promising alternative, and Bacillus thuringiensis endotoxin genes have played a major role in this respect. However, to overcome insect tolerance issues and to broaden the target range, it is critical to identify alternative-insecticidal toxins working through novel mechanisms. Our research group has identified a kinase from Chilo iridescent virus (CIV; Family Iridoviridae) that has insecticidal activity and designated it as ISTK (Iridovirus Serine/Threonine Kinase). A 35 kDa truncated form of ISTK, designated iridoptin, was obtained during expression and purification of ISTK in the yeast system. This yeast-expressed CIV toxin induced 50% mortality in cotton aphids and 100% mortality in green peach aphids (GPA). Optimized viral genes (o-ISTK and o-IRI) were stably transformed into the model plant, Arabidopsis. PCR analysis of genomic DNA confirmed the presence of the gene insert (oISTK/oIRI) in selected transgenic lines. The further screening was performed to identify the PCR positive lines that showed expression of respective toxins at the polypeptide level using Western blot analysis. The stable lines expressing either of these two toxins induced moderate to very high mortality in GPAs and significantly affected GPA development and fecundity. The aphicidal potential of these transgenic Arabidopsis lines will be presented.

Keywords: Chilo iridescent virus, insecticidal toxin, iridoviruses, plant-incorporated protectants, serine/threonine kinase

Procedia PDF Downloads 259

Authors: Genesis Julyus T. Agcaoili, Esperanza C. Cabrera

Abstract:

Thirty (30) isolates of lactic acid bacteria (LAB) from traditionally-prepared fermented products specifically fermented soy-bean paste, fermented mustard and fermented rice-fish mixture were studied for their in vitro antimicrobial and cytotoxic activities. Seventeen (17) isolates were identified as Lactobacillus plantarum, while 13 isolates were identified as Enterococcus spp using 16s rDNA sequences. Disc diffusion method was used to determine the antibacterial activity of LAB against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), while the modified agar overlay method was used to determine the antifungal activity of LAB isolates on the yeast Candida albicans, and the dermatophytes Microsporum gypseum, Trichophyton rubrum and Epidermophyton floccosum. The filter-sterilized LAB supernatants were evaluated for their cytotoxicity to mammalian colon cancer cell lines (HT-29 and HCT116) and normal human dermal fibrolasts (HDFn) using resazurin assay (PrestoBlueTM). Colchicine was the positive control. No antimicrobial activity was observed against the bacterial test organisms and the yeast Candida albicans. On the other hand, all of the tested LAB strains were fungicidal for all the test dermatophytes. Cytotoxicity index profiles of the supernatants of the 15 randomly picked LABs and negative control (brain heart infussion broth) suggest nontoxicity to the cells when compared to colchicine, whereas all LAB supernatants were found to be cytotoxic to HT-29 and HCT116 colon cancer cell lines. Results provide strong support for the role of the lactic acid bacteria studied in antimicrobial treatment and anticancer therapy.

Keywords: antimicrobial, fermented products, fungicidal activity, lactic acid bacteria, probiotics

Procedia PDF Downloads 218

Authors: Dupe Olufunke Ogunbosoye, Thaofik Badmos Mustapha, Lanre Shaffihy Adeaga, R. O. Imam

Abstract:

Efforts have been made to reduce ruminants' methane emissions while improving animal productivity. Hence, an experiment was conducted to investigate the in vitro fermentation pattern, methane production, and performance of West African dwarf (WAD) rams-fed diets at graded levels of ginger. Sixteen (16) rams were randomly allocated into four dietary treatments with four animals per treatment in a completely randomized design for 84 days. Ginger powder was added at 0.00%, 0.25%, 0.50% and 0.75% as T1, T2, T3 and T4 respectively. The results indicated that at the 24-hour diet incubation, gas production, methane, metabolizable energy (ME), organic matter digestibility (OMD), and short-chain fatty acids (SCFA) concentrations decreased with the increasing level of ginger. Conversely, the sheep-fed T4 recorded the highest daily weight gain (47.61g/day), while the least daily weight gain (17.86g/day) was recorded in ram-fed T1. The daily weight gain of the rams fed T3 and T4 was similar but significantly different from the daily weight gain in T1 (17.86g/day) and T2 (29.76g/day). Daily feed intake was not significantly different across the treatments. T4 recorded the best response regarding feed conversion ratio (18.59) compared with other treatments. Based on the results obtained, rams fed T4 perform best in terms of growth and methane production. It is therefore concluded that the addition of ginger powder into the diet of sheep up to 0.75% enhances the growth rate of WAD sheep and reduces enteric methane production to create a smart nutrition system in ruminant animal production.

Keywords: enteric methane, growth, in vitro, sheep, nutrition system

Procedia PDF Downloads 58

Authors: Sara Franco Ortega, Alina Goldberg Cavalleri, Nawaporn Onkokesung, Richard Dale, Melissa Brazier-Hicks, Robert Edwards

Abstract:

Safeners are agrochemicals that enhance the selective chemical control of wild grasses by increasing the ability of the crop to metabolise the herbicide. Although these compounds are widely used, their mode of action is not well understood. It is known that safeners enhance the metabolism of herbicides, by up-regulating the associated detoxification system we have termed the xenome. The xenome proteins involved in herbicide metabolism have been previously divided into four different phases, with cytochrome P450s (CYPs) playing a key role in phase I metabolism by catalysing hydroxylation and dealkylation reactions. Subsequently, glutathione S-transferases (GSTs) and UDP-glucosyltransferases lead to the formation of Phase II conjugates prior to their transport into the vacuole by ABCs transporters (Phase III). Maize (Zea mays), was been treated with different safeners to explore the selective induction of xenome proteins, with a special interest in the regulation of the CYP superfamily. Transcriptome analysis enabled the identification of key safener-inducible CYPs that were then functionally assessed to determine their role in herbicide detoxification. In order to do that, CYP’s were codon optimised, synthesised and inserted into the yeast expression vector pYES3 using in-fusion cloning. CYP’s expressed as recombinant proteins in a strain of yeast engineered to contain the P450 co-enzyme (cytochrome P450 reductase) from Arabidopsis. Microsomes were extracted and treated with herbicides of different chemical classes in the presence of the cofactor NADPH. The reaction products were then analysed by LCMS to identify any herbicide metabolites. The results of these studies will be presented with the key CYPs identified in maize used as the starting point to find orthologs in other crops and weeds to better understand their roles in herbicide selectivity and safening.

Keywords: CYPs, herbicide detoxification, LCMS, RNA-Seq, safeners

Procedia PDF Downloads 116

Authors: T. I. Mbata, M. J. Ikenebomeh

Abstract:

Background: The study was aim at formulating a complementary foods based on maize and bambara groundnut with a view of reducing malnutrition in low income families. Protein-energy malnutrition is a major health challenge attributed to the inappropriate complementary feeding practices, low nutritional quality of traditional complementary foods and high cost of quality protein-based complementary foods. Methods: The blends 70% maize, 30% bambara groundnut were evaluated for proximate analyses, minerals, amino acids profile, and antinutritional factors, using proprietary formula (‘Nutrend’) as standard. Antinutritional factors, amino acids, microbiological properties and sensory attributes were determined using standard methods. Results; For Protein, the results were 15.0% for roasted bambara groundnut maize germinated flour (RBMGF), 13.80% for cooked bambara groundnut maize germinated flour (CBMGF), 15.18% for soaked bambara groundnut maize germinated flour (SBMGF); values for maize flour and nutrend had 10.4% and 23.21% respectively. With respect to energy value, RBMGF, CBMGF, SBMGF, maize flour and nutrend had 494.9, 327.58, 356.49, 366.8 and 467.2 kcal respectively. The percentages of total essential amino acids in the composition of the blends were 36.9%, 40.7% and 38.9% for CBMGF, SBMGF and RBMGF, respectively, non-essential amino acids contents were 63.1%, 59.3% and 61.1% for CBMGF, SBMGF and RBMGF respectively. The mineral content, that is, calcium, potassium, magnesium and sodium, of formulated samples were higher than those obtained for maize flour and Nutrend. The antinutrient composition of RBMGF and CBMGF were lower than of SBMGF. The rats fed with the control diet exhibited better growth performance such as feed intake (1527 g) and body weight gain (93.8 g). For the microbial status, microflora gradually changed from gram negative enteric bacteria, molds, lactic acid bacteria and yeast to be dominated by gram positive lactic acid bacteria (LAB) and yeasts. Yeasts and LAB growth counts in the complementary food varied between 4.44 and 7.36 log cfu/ml. LAB number increased from 5.40 to 7.36 log cfu/ml during fermentation. Yeasts increased from 4.44 to 5.60 log cfu/ml. Organoleptic evaluation revealed that the foods were well accepted. Conclusion: Based on the findings the application of bambara groundnut fortification to traditional foods can promote the nutritional quality of African maize - based traditional foods with acceptable rheological and cooking qualities.

Keywords: bambara groundnut, maize, fortification, complementary food

Procedia PDF Downloads 338

Authors: Jyosthna Khanna Goli, Shaik Naseeruddin, Hameeda Bee

Abstract:

Employing lignocellulosic biomass in fermentative production of xylitol and bioethanol is gaining interest as it is renewable, cheap, and abundantly available. Xylitol is a polyol, gaining its importance in the food and pharmacological industry due to its low calorific value and anti-cariogenic nature. Bioethanol from lignocellulosic biomass is widely accepted as an alternative fuel for transportation with reduced CO₂ emissions, thus reducing the greenhouse effect. Typha latifolia, an aquatic weed, was found to be promising lignocellulosic substrate as it posses a high amount of sugars and does not compete with arable lands and interfere with food and feed competition. In the present study, xylose from hemicellulosic fraction of typha is converted to xylitol by isolate Jfh5 (Candida. tropicalis) and cellulose part to ethanol using Saccharomyces cerevisiaeVS3. Initially, alkali pretreatment of typha using sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, sodium bisulphate and sodium dithionate for overnight (18h) at room temperature (28 ± 2°C), resulted in maximum delignification of 75% with 2% (v/v) sodium bisulphate. Later, pretreated biomass was subjected to acid hydrolysis with 1%, 1.5%, 2%, and 3% H₂SO₄ at 110 °C and 121°C for 30 and 60 min, respectively. 2% H₂SO₄ at 121°C for 60 min was found to release 13.5 g /l sugars, which on detoxification and fermentation produced 8.1g/l xylitol with yield and productivity of 0.65g/g and 0.112g/l/h respectively. Further enzymatic hydrolysis of the residual substrate obtained after acid hydrolysis released 11g/l sugar, which on fermentation with VS3 produced 4.9g/l ethanol with yield and productivity of 0.22g/g and 0.136g/l/h respectively.

Keywords: delignification, xylitol, bioethanol, acid hydrolysis, enzyme hydrolysis

Procedia PDF Downloads 130

Authors: Mirosław Krzemieniewski, Marcin Zieliński, Marcin Dębowski

Abstract:

Electrokinetic disintegration is one of the high-voltage electric methods. The design of systems is exceptionally simple. Biomass flows through a system of pipes with alongside mounted electrodes that generate an electric field. Discharges in the electric field deform cell walls and lead to their successive perforation, thereby making their contents easily available to bacteria. The spark-over occurs between electrode surface and pipe jacket which is the second pole and closes the circuit. The value of voltage ranges from 10 to 100kV. Electrodes are supplied by normal “power grid” monophase electric current (230V, 50Hz). Next, the electric current changes into direct current of 24V in modules serving for particular electrodes, and this current directly feeds the electrodes. The installation is completely safe because the value of generated current does not exceed 250mA and because conductors are grounded. Therefore, there is no risk of electric shock posed to the personnel, even in the case of failure or incorrect connection. Low values of the electric current mean small energy consumption by the electrode which is extremely low – only 35W per electrode – compared to other methods of disintegration. Pipes with electrodes with diameter of DN150 are made of acid-proof steel and connected from both sides with 90º elbows ended with flanges. The available S and U types of pipes enable very convenient fitting with system construction in the existing installations and rooms or facilitate space management in new applications. The system of pipes for electrokinetic disintegration may be installed horizontally, vertically, askew, on special stands or also directly on the wall of a room. The number of pipes and electrodes is determined by operating conditions as well as the quantity of substrate, type of biomass, content of dry matter, method of disintegration (single or circulatory), mounting site etc. The most effective method involves pre-treatment of substrate that may be pumped through the disintegration system on the way to the fermentation tank or recirculated in a buffered intermediate tank (substrate mixing tank). Biomass structure destruction in the process of electrokinetic disintegration causes shortening of substrate retention time in the tank and acceleration of biogas production. A significant intensification of the fermentation process was observed in the systems operating in the technical scale, with the greatest increase in biogas production reaching 18%. The secondary, but highly significant for the energetic balance, effect is a tangible decrease of energy input by agitators in tanks. It is due to reduced viscosity of the biomass after disintegration, and may result in energy savings reaching even 20-30% of the earlier noted consumption. Other observed phenomena include reduction in the layer of surface scum, reduced sewage capability for foaming and successive decrease in the quantity of bottom sludge banks. Considering the above, the system for electrokinetic disintegration seems a very interesting and valuable solutions meeting the offer of specialist equipment for the processing of plant biomass, including Virginia fanpetals, before the process of methane fermentation.

Keywords: electrokinetic disintegration, biomass, biogas production, fermentation, Virginia fanpetals

Procedia PDF Downloads 349

Authors: G. J. Fan, T. T. Lee, M. H. Chen, T. F. Shiao, B. Yu, C. F. Lee

Abstract:

Rice straw with poor feed quality and spent mushroom substrate are both the most abundant agricultural residues in Taiwan. Edible mushrooms from white rot fungi possess lignocellulase activity. It was expected to improve the feeding value of rice straw for ruminant by solid-state fermentation pretreatment using spent mushroom substrate. Six varieties or subspecies of spent edible mushrooms (Pleurotus ostreatus (blue or white color), P. sajor-caju, P. citrinopileatus, P. eryngii and Ganoderma lucidium) substrate were evaluated in solid-state fermentation process with rice straw for 8 wks. Quality improvement of fermented rice straw was determined by its in vitro digestibility, lignocellulose degradability, and cell wall breakdown checked by scanning electron microscope. Results turned out that Pleurotus ostreatus (white color) and P. sajor-caju had the better lignocellulose degradation effect than the others and was chosen for advance in vivo study. Rice straw fermented with spent Pleurotus ostreatus or Pleurotus sajor-caju mushroom substrate 8 wks was prepared for growing and lactating feeding trials of dairy goat, respectively. Pangolagrass hay at 15% diet dry matter was the control diet. Fermented or original rice straw was added to substitute pangolagrass hay in both feeding trials. A total of 30 head of Alpine castrated ram were assigned into three groups for 11 weeks, 5 pens (2 head/pen) each group. A total of 21 head of Saanen and Alpine goats were assigned into three treatments and individually fed in two repeat lactating trials with 28-d each. In castrated ram study, results showed that fermented rice straw by spent Pleurotus ostreatus mushroom substrate attributed the higher daily dry matter intakes (DMI, 1.53 vs. 1.20 kg) and body weight gain (138 vs. 101 g) than goats fed original rice straw. DMI (2.25 vs. 1.81 kg) and milk yield (3.31 vs. 3.02 kg) of lactating goats fed control pangolagrass diet and fermented rice straw by spent Pleurotus sajor-caju mushroom substrate were also higher than those fed original rice straw diet (P < 0.05). Milk compositions, milk fat, protein, total solid and lactose, were similar among treatments. In conclusion, solid-state fermentation by spent Pleurotus ostreatus or Pleurotus sajor-caju mushroom substrate could effectively improve the feeding value of rice straw. Fermented rice straw is a good alternative fiber feed resource for growing and lactating dairy goats and 15% in diet dry matter is recommended.

Keywords: feeding value, fermented rice straw, growing and lactating dairy goat, spent Pleurotus ostreatus and Pleurotus sajor-caju mushroom substrate

Procedia PDF Downloads 153

Authors: Ilirjan Malollari, Redi Buzo, Lorina Lici

Abstract:

This study was done for the characterization of polyphenols changes of anthocyanins, flavonoids, the color intensity and total polyphenols index, maturity and oxidation index during the process of micro-oxygenation of wine that comes from a specific geographic area in the southeastern region of the country. Also, through mathematical modeling of the oxygen distribution within solution of wort for wine fermentation, was shown the strong impact of carbon dioxide present in the liquor. Analytical results show periodic increases of color intensity and tonality, reduction level of free anthocyanins and flavonoids free because of polycondensation reactions between tannins and anthocyanins, increased total polyphenols index and decrease the ratio between the flavonoids and anthocyanins offering a red stabilize wine proved by sensory degustation tasting for color intensity, tonality, body, tannic perception, taste and remained back taste which comes by specific area associated with environmental indications. Micro-oxygenation of wine is a wine-making technique, which consists in the addition of small and controlled amounts of oxygen in the different stages of wine production but more efficiently after end of alcoholic fermentation. The objectives of the process include improved mouth feel (body and texture), color enhanced stability, increased oxidative stability, and decreased vegetative aroma during polyphenols changes process. A very important factor is polyphenolics organic grape composition strongly associated with the environment geographical specifics area in which it is grown the grape.

Keywords: micro oxygenation, polyphenols, environment, wine stability, diffusion modeling

Procedia PDF Downloads 192

Authors: Elif Coskun Daggecen, Seyma Dokucu, Yekta Gezginc, Ismail Akyol

Abstract:

Fermented dairy food quality is mainly determined by the sensory perception and influenced by many factors. Today, starter cultures for fermented foods are being developed to have a constant quality in these foods. Streptococcus thermophilus is one of the main species of most a starter cultures of yogurt fermentation. This species produces lactate by lactose fermentation from pyruvate. On the other hand, a small amount of pyruvate can alternatively be converted to various typical yoghurt flavor compounds such as diacetyl, acetoin, acetaldehyde, or acetic acid, for which the activity of three genes are shown to be especially important; ldh, nox and als. Up to date, commercially produced yoghurts have not yet met the desired aromatic properties that Turkish consumers find in traditional homemade yoghurts. Therefore, it is important to select starters carrying favorable metabolic characteristics from natural isolates. In this study, 30 strains of Str. Thermophilus were isolated from traditional Turkish yoghurts obtained from different regions of the country. In these strains, transcriptional levels of ldh, nox and als genes were determined via a newly developed qPCR protocol, which is a more reliable and precision method for analyzing the quantitative and qualitative expression of specific genes in different experimental conditions or in different organisms compared to conventional analytical methods. Additionally, the metabolite production potentials of the isolates were measured. Of all the strains examined, 60% were found to carry the metabolite production potential and the gene activity which appeared to be suitable to be used as a starter culture. Probable starter cultures were determined according to real-time PCR results.

Keywords: gene expression, RT-PCR, starter culture, Streptococcus thermophilus

Procedia PDF Downloads 168

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

Abstract:

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

Procedia PDF Downloads 125

Authors: Idan Chiyanzu, Maruping Mangena

Abstract:

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 264

Authors: Agumah N. B. Orji, M. U., Oru C. M., Ugbo, E. N., Onwuliri E. A Nwakaeze, E. A.,

Abstract:

ERG11 gene has been reported to be one of the genes whose expression is responsible for resistance of Candida to various triazole drugs, which are first line treatment for candidiasis. This study was carried out to determine the prevalence of Triazole (Fluconazole and voriconazole) resistant Candida albicans expressing ERG11 gene from adult females in Abakaliki. Urine and vaginal swab samples were randomly collected from volunteers after obtaining their consent to participate in the study. A total of 565 adult females participated in the study. A total of 340 urine specimens and 288 vaginal swab specimens were collected. Direct wet mount technique, as well as culture in Trichomonas broth, were used to examine the urine and vaginal swab specimens for the presence of motile Trichomonads. The Trichomonas broth used was selective for both T. vaginalis and C. albicans. Broths that yielded budding yeast cells after microscopy were subcultured on to Sabouraud dextrose agar, after which Germ tube test was carried out to confirm the presence of C. albicans. Biochemical tests, including carbohydrate fermentation and urease utilization, were also performed. Antibiogram of C. albicans isolates obtained from this study was carried out using commercially available azole drugs. Fluconazole and voriconazole were selected as Triazole drugs used for this study. Nystatin was used as a tangential control. An MIC test was carried out with E-strips on some of the resistant C. albicans isolates A total of 6 isolates that resisted all the azole drugs were selected and screened for the presence of ERG11 gene using Reverse transcriptase polymerase chain reaction technique. The total prevalence recorded for C. albicans was 13.0%. Frequency was statistically higher in Pregnant (7.96%) than non pregnant (5.09%) volunteers (X2=0.94 at P=0.05). With respect to clinical samples, frequency was higher in vaginal swabs samples (7.96%) than Urine samples (5.09%) (X2=9.05 at P=0.05). Volunteers within the age group 26-30 years recorded the highest prevalence (4.46%), while those within the age group 36-40 years recorded the lowest at 1.27%(X2=4.34 at P=0.05). In pregnant female participants, the highest frequency was recorded with those in their 3rd trimester (4.14%), while lowest incidence was recorded for those in their first trimester (0.80%). Antibiogram results from this study showed that C. albicans isolates obtained from this study resisted Fluconazole (72%) more than Voriconazole (57%). Only one out of the six selected isolates yielded resistance in the MIC test. Results obtained from the RT-PCR showed that there was no expression of ERG11 gene among the fluconazole resistant isolates of C. albicans. Observed resistance may be due to other factors other than expression of ERG11 gene.

Keywords: candida, ERG11, triazole, nigeria

Procedia PDF Downloads 131

Authors: Mardiati Zain, W. S. N. Rusmana, Erpomen, Malik Makmur, Ezi Masdia Putri

Abstract:

Background and Aim: The leaves of the Leucaenaleucocephala tree have potential as a nitrogen source for ruminants. Leucaena leaf meal as protein supplement has been shown to improve the feed quality of ruminants. The effects of different levels of Leucaena leucocephala supplementation as substitute of concentrate on fiber digestibility and in vitro rumen fermentation characteristics were investigated. This research was conducted in vitro. The study used a randomized block design consisting of 3 treatments and 5 replications. The treatments were A. 40% rice straw ammoniated + 60% concentrate, B. 40% rice straw ammoniated + 50% concentrate + 10% Leucaena leuchephala, C. 40% rice straw ammoniated + 40% concentrate + 20% Leucaena leuchephala, Result: The results showed that the addition of Leucaena leucocephala increased the digestibility of Neutral detergent Fiber NDF and Acid Detergent Fiber (ADF) (p < 0.05). In this study, rumen NH3, propionate, amount of escape protein and total Volatyl Fatty Acid (VFA) were found increased significantly at treatment B. No significant difference was observed in acetate and butyrate production. The populations of total protozoa and methane production had significantly decreased (P < .05) in supplemented group. Conclusion: Supplementation of leuchaena leucochepala on completed feed based on ammoniated rice straw in vitro can increase fiber digestibility, VFA production and decreased protozoa pupulataion and methane production. Supplementation of 10% and 20% L. leucochepala were suitable to be used for further studies, therefore in vivo experiment is required to study the effects on animal production.

Keywords: digestibility, Leucaena leucocephala, complete feed, rice straw ammoniated

Procedia PDF Downloads 133

Authors: Philip Vercoe, Ali Hardan

Abstract:

Samples of 29 Australian wild natives and exotic plants were tested in vitro batch rumen culture system for their methanogenic characteristics and potential usage as feed or antimicrobial to enhance sustainable livestock ruminant production system. The plants were tested for their in vitro rumen fermentation end products properties which include: methane production, total gas pressure, concentrations of total volatile fatty acids, ammonia, and acetate to propionate ratio. All of the plants were produced less methane than the positive control (i.e., oaten chaff) in vitro. Nearly 50 % of plants inhibiting methane by over 50% in comparison to the control. Eremophila granitica had the strongest inhibitory effect about 92 % on methane production comparing with oaten chaff. The exotic weed Arctotheca calendula (Capeweed) had the highest concentration of volatile fatty acids production as well as the highest in total gas pressure among all plants and the control. Some of the acacia species have the lowest production of total gas pressure. The majority of the plants produced more ammonia than the oaten chaff control. The plant species that produced the most ammonia was Codonocarpus cotinifolius, producing over 3 times as much methane as oaten chaff control while the lowest was Eremophila galeata. There was strong positive correlation between methane production and total gas production as well as between total gas production and the concentration of VFA produced with R² = 0.74, R² = 0.84, respectively. While there was weak positive correlation between methane production and the acetate to propionate ratio as well as between the concentration of VFA produced and methane production with R² = 0.41, R² = 0.52, respectively.

Keywords: in vitro Rumen Fermentation, methane, wild Australian native plants, forages

Procedia PDF Downloads 325

Authors: Abu Yousuf, Maksudur Rahman Khan, Ahasanul Karim, Amirul Islam, Minhaj Uddin Monir, Sharmin Sultana, Domenico Pirozzi

Abstract:

Traditional biodiesel feedstock like edible oils or plant oils, animal fats and cooking waste oil have been replaced by microbial oil in recent research of biodiesel synthesis. The well-known community of microbial oil producers includes microalgae, oleaginous yeast and seaweeds. Conventional transesterification of microbial oil to produce biodiesel is lethargic, energy consuming, cost-ineffective and environmentally unhealthy. This process follows several steps such as microbial biomass drying, cell disruption, oil extraction, solvent recovery, oil separation and transesterification. Therefore, direct transesterification of biodiesel synthesis has been studying for last few years. It combines all the steps in a single reactor and it eliminates the steps of biomass drying, oil extraction and separation from solvent. Apparently, it seems to be cost-effective and faster process but number of difficulties need to be solved to make it large scale applicable. The main challenges are microbial cell disruption in bulk volume and make faster the esterification reaction, because water contents of the medium sluggish the reaction rate. Several methods have been proposed but none of them is up to the level to implement in large scale. It is still a great challenge to extract maximum lipid from microbial cells (yeast, fungi, algae) investing minimum energy. Electroporation technique results a significant increase in cell conductivity and permeability caused due to the application of an external electric field. Electroporation is required to alter the size and structure of the cells to increase their porosity as well as to disrupt the microbial cell walls within few seconds to leak out the intracellular lipid to the solution. Therefore, incorporation of electroporation techniques contributed in direct transesterification of microbial lipids by increasing the efficiency of biodiesel production rate.

Keywords: biodiesel, electroporation, microbial lipids, transesterification

Procedia PDF Downloads 258

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

Abstract:

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

Procedia PDF Downloads 313

Authors: Carol N. Flores-Fernández, Guadalupe Espilco, Cynthia Esquerre, Amparo I. Zavaleta

Abstract:

Saline environments represent a valuable source of enzymes with novel properties and particular features for application in food, pharmaceutical and chemical industry. This study focuses on the isolation and screening of hydrolase-producing bacteria from Chilca salterns and the evaluation of their biotechnological potential. Soil samples were collected from Chilca salterns in Peru. For the isolation, medium containing 0.2 % of yeast extract, 5 % of NaCl and 10 % of the soil sample was used. After 72 h of incubation at 37 °C, serial dilutions were made up to 10−12 dilutions, spread on agar plates with 0.5 % of yeast extract and 5 % of NaCl, and incubated at 37 °C for 48 h. Screening of hydrolase-producing bacteria was carried out for cellulases, amylases, lipases, DNase, and proteases on specific media. Moreover, protease-producing bacteria were tested using protein extracted from the following legumes as substrate: Glycine max, Lupinus mutabilis, Pisum sativum, Erythrina edulis, Cicer arietinum, Phaseolus vulgaris and Vicia faba. A total of 16 strains were isolated from soil samples. On the screening media; 75, 44, 81 and 50 % were cellulase, amylase, DNase and protease producers, respectively. Also, 19 % of the isolates produced all the hydrolytic enzymes above mentioned. Lipase producers were not found. The 37 % and 12 % of the strains grew at 20 % and 30 % of salt concentration, respectively. In addition, 75 % of the strains grew at pH range between 5 and 10. From the total of protease-producing bacteria, 100 % hydrolyzed Glycine max, Lupinus mutabilis, and Pisum sativum protein, while 87 % hydrolyzed Erythrina edulis and Cicer arietinum protein. Finally, 75 % and 50 % of the strains hydrolyzed Phaseolus vulgaris and Vicia faba protein, respectively. Hydrolase-producing bacteria isolated from Chilca salterns in Peru grew at high salt concentrations and wide range of pH. In addition, protease-producing bacteria hydrolyzed protein from different sources such as leguminous. These enzymes have great biotechnological potential and could be used for different industrial processes and applications.

Keywords: bacteria, extracellular, hydrolases, Peru, salterns

Procedia PDF Downloads 182

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

Abstract:

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 279

Authors: M. C. Muya, L. J. Erasmus

Abstract:

Twenty four new born male Holstein calves were divided into two treatments groups and used to evaluate the effects of M. elsdenii NCIMB 41125. The first groups were dosed with 50 ml containing 108 CFU/mL of M. elsdenii NCIMB 41125 (Me) and the control calves were not dosed. Within each of the two treatments groups, calves were divided into three treatment groups (Not dosed: 7 d, 14 d and 21 d vs dosed Me 7 d, Me14 and Me21 d (treatments), each groups contained 4 calves within which two calves were euthanized at 24 h and two calves at 72 h. Calves entered the trial until euthanize at whether 24 or 72 H after dosing time. After receiving colostrum for 3 consecutive days after birth, calves were fed whole milk and had free access to a commercial calf starter pellet and fresh water. Fecal grab samples were taken from each calf in duplicate +24 h or +72 h relative to dosing. Immediately after euthanizing, the digestive tract was harvested, and duplicate rumen and colon digesta samples collected for VFA’s determination and DNA extraction for bacteria count using 16s RNA PCR probe technique. Independent two t-test was performed to compare mean volatile fatty acids. Mixed-effects linear regressions were performed to establish relationships between: 1) M. elsdenii and Me, and between VFA’s and Me using SAS (2009). M. elsdenii NCIMB 41125 was detected in the faeces, colon and rumen of dosed calves at both +24H and +72H and ranged from 1.6 x 106 to 4.9 x 109 cfu/ml, indicating its potential to colonize in the digestive tract of calves. There was a strong positive relationship (R²=0.96; P < 0.0001) between M. elsdenii NCIMB 41125 and M. elsdenii population (cfu/ml) in the rumen, suggesting that the increase in M. elsdenii was due to increased M. elsdenii NCIMB 41125. An increase in butyrate was observed from +24 h to +72 h when calves were dosed on both d 7 and 14. Results showed that Me presented a positive relationship with butyrate (P < 0.001, R² = 0.43) and a concomitant negative relationship with acetate (P = 0.017, R² = -0.33). These results suggest that dosing pre-weaned dairy calves with M. elsdenii NCIMB 41125 has the potential to alter ruminal VFA production through increasing proportions of butyrate at the expense of propionate.

Keywords: calves, megasphaera elsdenii, rumen fermentation, bacteria

Procedia PDF Downloads 373

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

Abstract:

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

Procedia PDF Downloads 122

Authors: Chang-Hui Shen, Michelle Esposito, Andrew J. Shen, Michael Adejokun, Diana Laterman

Abstract:

The packaging of DNA into nucleosomes is critical to genomic compaction, yet it can leave gene promoters inaccessible to activator proteins or transcription machinery and thus prevents transcriptional initiation. Both chromatin remodelers and histone acetylases (HATs) are the two main transcription co-activators that can reconfigure chromatin structure for transcriptional activation. Ino80p is the core component of the INO80 remodeling complex. Recently, it was shown that Ino80p dissociates from the yeast INO1 promoter after induction. However, when certain HATs were deleted or mutated, Ino80p accumulated at the promoters during gene activation. This suggests a link between HATs’ presence and Ino80p’s dissociation. However, it has yet to be demonstrated that Ino80p can be acetylated. To determine if Ino80p can be acetylated, wild-type Saccharomyces cerevisiae cells carrying Ino80p engineered with a double FLAG tag (MATa INO80-FLAG his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) were grown to mid log phase, as were non-tagged wild type (WT) (MATa his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) and ino80∆ (MATa ino80∆::TRP1 his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) cells as controls. Cells were harvested, and the cell lysates were subjected to immunoprecipitation (IP) with α-FLAG resin to isolate Ino80p. These eluted IP samples were subjected to SDS-PAGE and Western blot analysis. Subsequently, the blots were probed with the α-FLAG and α-acetyl lysine antibodies, respectively. For the blot probed with α-FLAG, one prominent band was shown in the INO80-FLAG cells, but no band was detected in the IP samples from the WT and ino80∆ cells. For the blot probed with the α-acetyl lysine antibody, we detected acetylated Ino80p in the INO80-FLAG strain while no bands were observed in the control strains. As such, our results showed that Ino80p can be acetylated. This acetylation can explain the co-activator’s recruitment patterns observed in current gene activation models. In yeast INO1, it has been shown that Ino80p is recruited to the promoter during repression, and then dissociates from the promoter once de-repression begins. Histone acetylases, on the other hand, have the opposite pattern of recruitment, as they have an increased presence at the promoter as INO1 de-repression commences. This Ino80p recruitment pattern significantly changes when HAT mutant strains are studied. It was observed that instead of dissociating, Ino80p accumulates at the promoter in the absence of functional HATs, such as Gcn5p or Esa1p, under de-repressing processes. As such, Ino80p acetylation may be required for its proper dissociation from the promoters. The remodelers’ dissociation mechanism may also have a wide range of implications with respect to transcriptional initiation, elongation, or even repression as it allows for increased spatial access to the promoter for the various transcription factors and regulators that need to bind in that region. Our findings here suggest a previously uncharacterized interaction between Ino80p and other co-activators recruited to promoters. As such, further analysis of Ino80p acetylation not only will provide insight into the role of epigenetic modifications in transcriptional activation, but also gives insight into the interactions occurring between co-activators at gene promoters during gene regulation.

Keywords: acetylation, chromatin remodeler, epigenetic modification, Ino80p

Procedia PDF Downloads 151

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

Abstract:

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

Procedia PDF Downloads 95

Authors: Amriana Hifizah, Philip Edward Vercoe, Graeme Bruce Martin, Teuku Reza Ferasy, Muhammad Hambal

Abstract:

Many native forage plant species have been used in Indonesia as feed for ruminants. However, less information is available about how these plants affect productivity, let alone methane emissions. In the province of Aceh, where the traditional practice is to feed local forages to small ruminants, the farmers are not satisfied with the productivity of their livestock, and they attribute this problem to poor availability and too few options for good quality forages. Forage quality is reduced by high environmental temperatures which increase the amount of lignification. In addition to reducing productivity, these factors also increase enteric methane production. A preliminary survey about potential forage species was completed in three different districts, two of low elevation and one of high elevation: Syiah Kuala (05°30’5.08” N to 095°24’7.35” E), elevation 29 m MSL; Kajhu (05°32’34.6” N to 095°21’17.7” E), elevation 30 m MSL; Lembah Seulawah (05°28'06.4" N to 095°43' 14.2" E), elevation 254 m MSL. Information about local plants was collected in a semi-structured interview with scientists, government field officers and local farmers, in the city of Banda Aceh and in those three districts. The outcome was a list 40 species that could be useful, of which 21 were selected for further study. The selection process was based on several criteria: high availability, high protein content, low toxicity, and evidence of secondary metabolites (eg, history of medicinal plants for both human and animals). For some of the selected medicinal plants, there is experimental evidence of effects on methane production during rumen fermentation. Subsequently, the selected forages were tested for their effects on rumen fermentation in vitro, using batch culture. The data produced will be used to identify forages with the potential to reduce CH4 emissions. These candidates will then be assessed for their benefits (fermentability and productivity) and potential deleterious side-effects.

Keywords: batch culture, forage, methane, rumen

Procedia PDF Downloads 312