Search results for: enzymatic digestibility

Authors: Maxime Merheb, Rachel Matar

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Retrieving ancient DNA sequences which in return permit the entire genome sequencing from fossils have extraordinarily improved in recent years, thanks to sequencing technology and other methodological advances. In any case, the quest to search for ancient DNA is still obstructed by the damage inflicted on DNA which accumulates after the death of a living organism. We can characterize this damage into three main categories: (i) Physical abnormalities such as strand breaks which lead to the presence of short DNA fragments. (ii) Modified bases (mainly cytosine deamination) which cause errors in the sequence due to an incorporation of a false nucleotide during DNA amplification. (iii) DNA modifications referred to as blocking lesions, will halt the PCR extension which in return will also affect the amplification and sequencing process. We can clearly see that the issues arising from breakage and coding errors were significantly decreased in recent years. Fast sequencing of short DNA fragments was empowered by platforms for high-throughput sequencing, most of the coding errors were uncovered to be the consequences of cytosine deamination which can be easily removed from the DNA using enzymatic treatment. The methodology to repair DNA sequences is still in development, it can be basically explained by the process of reintroducing cytosine rather than uracil. This technique is thus restricted to amplified DNA molecules. To eliminate any type of damage (particularly those that block PCR) is a process still pending the complete repair methodologies; DNA detection right after extraction is highly needed. Before using any resources into extensive, unreasonable and uncertain repair techniques, it is vital to distinguish between two possible hypotheses; (i) DNA is none existent to be amplified to begin with therefore completely un-repairable, (ii) the DNA is refractory to PCR and it is worth to be repaired and amplified. Hence, it is extremely important to develop a non-enzymatic technique to detect the most degraded DNA.

Keywords: ancient DNA, DNA barcodong, enzymatic repair, PCR

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Authors: T. Vijay Bhasker, N. K. S Gowda, P. Krishnamoorthy, D. T. Pal, A. K. Pattanaik, A. K. Verma

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Boron (B) is a newer trace element and its biological importance and dietary essentiality is unclear in animals. The available literature suggests its putative role in bone mineralization, antioxidant status and steroid hormone synthesis. A feeding trial was conducted in Wister strain (Rattus norvegicus) albino rats for duration of 90 days. A total of 84 healthy weaned (3-4 weeks) experimental rats were randomly divided into 7 dietary groups (4 replicates of three each) viz., A (Basal diet/ Control), B (Basal diet + 5 ppm B), C (Basal diet + 10 ppm B), D (Basal diet + 20 ppm B), E (Basal diet + 40 ppm B), F (Basal diet-Ca 50%), G (Basal diet-Ca 50% + 40 ppm B). Dietary level of calcium (Ca) was maintained at two levels, 100% and 50% of requirement. Sodium borate was used as source of boron along with other ingredients of basal diet while preparing the pelletized diets. All the rats were kept in proper ventilated laboratory animal house maintained at temperature (23±2º C) and humidity (50 to 70%). At the end of experiment digestibility trial was conducted for 5 days to estimate nutrient digestibility and gut absorption of minerals. Eight rats from each group were sacrificed to collect the vital organs (liver, kidney and spleen) to study histopathology. Blood sample was drawn by heart puncture to determine biochemical profile. The average daily feed intake (g/rat/day), water intake (ml/rat/day) and body weight gain (g/rat/day) were similar among the dietary groups. The digestibility (%) of organic matter and crude fat were significantly improved (P < 0.05) was by B supplementation. The gut absorption (%) Ca was significantly increased (P < 0.01) in B supplemented groups compared to control. However, digestibility of dry matter and crude protein, gut absorption of magnesium and phosphorus showed a non-significant increasing trend with B supplementation. The gut absorption (%) of B (P < 0.01) was significantly lowered (P<0.05) in supplemented groups compared to un-supplemented ones. The serum level of triglycerides (mg/dL), HDL-cholesterol (mg/dL) and alanine transaminase (IU/L) were significantly lowered (P < 0.05) in B supplemented groups. While serum level of glucose (mg/dL) and alkaline phosphatase (KA units) showed a non-significant decreasing trend with B supplementation. However the serum levels of total cholesterol (mg/dL) and aspartate transaminase (IU/L) were similar among dietary groups. The histology sections of kidney and spleen revealed no significant changes among the dietary groups and were observed to be normal in anatomical architecture. However, the liver histology revealed cell degenerative changes with vacuolar degeneration and nuclear condensation in Ca deficient groups. But the comparative degenerative changes were mild in 40 ppm B supplemented Ca deficient group. In conclusion, dietary supplementation of graded levels of boron in rats had a positive effect on metabolism and health by improving nutrient digestibility and gut absorption of Ca. This indicates the beneficial role of dietary boron supplementation.

Keywords: boron, calcium, nutrient utilization, histopathology

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Authors: Mirnawati, Gita Ciptaan, Ferawati

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Background and Objective: Palm kernel cake (PKC) is a waste of the palm oil industry. Indonesia, as the largest palm oil producer in the world, produced 45-46% palm kernel cake. Palm kernel cake can potentially be used as animal ration but its utilization for poultry is limited. Thus, fermentation process was done in order to increase the utilization PKC in poultry ration. An experiment was conducted to study the effect between Inoculum Doses with Bacillus subtilis and fermentation time to improve the quality and nutrient content of fermented Palm Kernel Cake. Material and Methods: 1) Palm kernel cake derived from Palm Kernel Processing Manufacture of Andalas Agro Industry in Pasaman, West Sumatra. 2) Bacillus subtilis obtained from The Research Center of Applied Chemistry LIPI, Bogor. 3) Preparations nutrient agar medium (NA) produced by Difoo - Becton Dickinson. 4) Rice bran 5) Aquades and mineral standard. The experiment used completely randomize design (CRD) with 3 x 3 factorial and 3 replications. The first factors were three doses of inoculum Bacillus subtilis: (3%), (5%), and (7%). The second factor was fermentation time: (1) 2 day, (2) 4 day, and (3) 6 day. The parameters were crude protein, crude fiber, nitrogen retention, and crude fiber digestibility of fermented palm kernel cake (FPKC). Results: The result of the study showed that there was significant interaction (P<0.01) between factor A and factor B and each factor A and B also showed significant effect (P<0.01) on crude protein, crude fiber, nitrogen retention, and crude fiber digestibility. Conclusion: From this study, it can be concluded that fermented PKC with 7% doses of Bacillus subtilis and 6 days fermentation time provides the best result as seen from 24.65% crude protein, 17.35% crude fiber, 68.47% nitrogen retention, 53.25% crude fiber digestibility of fermented palm kernel cake (FPKC).

Keywords: fermentation, Bacillus Subtilis, inoculum, palm kernel cake, quality, nutrient

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Authors: Catherine Joke Adeseko

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Enzymatic browning is an economically important disorder that degrades organoleptic properties and prevent the consumer from purchasing fresh fruit and vegetables. Prevention and control of enzymatic browning in fruit and its product is imperative. Therefore, this study sought to investigate the catalytic effect of polyphenol oxidase (PPO) in the adverse browning of African bush mango (Irvingia gabonensis) fruit peel and pulp. PPO was isolated and purified, and its physicochemical properties, such as the effect of pH with SDS, temperature, and thermodynamic studies, which invariably led to thermal inactivation of purified PPO at 80 °C, were evaluated. The pH and temperature optima of PPO were found at 7.0 and 50, respectively. There was a gradual increase in the activity of PPO as the pH increases. However, the enzyme exhibited a higher activity at neutral pH 7.0, while enzymatic inhibition was observed at acidic region, pH 2.0. The presence of SDS at pH 5.0 downward was found to inhibit the activity of PPO from the peel and pulp of I. gabonensis. The average value of enthalpy (ΔH), entropy (ΔS), and Gibbs free energy (ΔG) obtained at 20 min of incubation and temperature 30 – 80 °C were respectively 39.93 kJ.mol-1, 431.57 J.mol-1 .K-1 and -107.99 kJ.mol-1 for peel PPO, and 37.92 kJ.mol-1, -442.51J.mol-1.K-1, and -107.22 kJ.mol-1 for pulp PPO. Thermal inactivation of PPO from I. gabonensis exhibited a reduction in catalytic activity as the temperature and duration of heat inactivation increases using catechol, reflected by an increment in k value. The half-life of PPO (t1/2) decreases as the incubation temperature increases due to the instability of the enzyme at high temperatures and was higher in pulp than peel. Both D and Z values decrease with increase in temperature. The information from this study suggests processing parameters for controlling PPO in the potential industrial application of I. gabonensis fruit in order to prolong the shelf-life of this fruit for maximum utilization.

Keywords: enzymatic, browning, characterization, activity

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Authors: Afshin Farahbakhsh, Hoda Khodadadi

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In this paper, we compare five methods of biological fuel cell fabrication by combining a Shewanella oneidensis microbial anode and a laccase-modified air-breathing cathode. As a result of biofuel cell laccase with graphite nanofibers, carbon surface (PAMAN) on the pt/hpg electrode, graphite sheets MWCNT and with (PG) and (MWCNT) showed, respectively. Describes methods for creating controllable and reproducible bio-anodes and demonstrates the versatility of hybrid biological fuel cells. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. When the device was fed with transdermal extracts, containing only 30μM of glucose, the average peak power was proportionally lower (0.004mW). The result of biofuel cell with graphite nanofibers showed the enzymatic fuel cell reaches 0.5 V at open circuit voltage with both, ethanol and methanol and the maximum current density observed for E2electrode was 228.94mAcm.

Keywords: enzymatic electrode, fuel cell, immobilization, laccase

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Authors: Anju Joshi, C. N. Tharamani

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Diabetes is one of the leading cause of death at present and remains an important concern as the prevalence of the disease is increasing at an alarming rate. Therefore, it is crucial to diagnose the accurate levels of glucose for developing an efficient therapeutic for diabetes. Due to the availability of convenient and compact self-testing, continuous monitoring of glucose is feasible nowadays. Enzyme based electrochemical sensing of glucose is quite popular because of its high selectivity but suffers from drawbacks like complicated purification and immobilization procedures, denaturation, high cost, and low sensitivity due to indirect electron transfer. Hence, designing a robust enzyme free platform using transition metal oxides remains crucial for the efficient and sensitive determination of glucose. In the present work, manganese doped nickel oxide nanoparticles (Mn-NiO) has been synthesized onto the surface of multiwalled carbon nanotubes using a simple microwave assisted approach for non-enzymatic electrochemical sensing of glucose. The morphology and structure of the synthesized nanostructures were characterized using scanning electron microscopy (SEM) and X-Ray diffraction (XRD). We demonstrate that the synthesized nanostructures show enormous potential for electrocatalytic oxidation of glucose with high sensitivity and selectivity. Cyclic voltammetry and square wave voltammetry studies suggest superior sensitivity and selectivity of Mn-NiO decorated carbon nanotubes towards the non-enzymatic determination of glucose. A linear response between the peak current and the concentration of glucose has been found to be in the concentration range of 0.01 μM- 10000 μM which suggests the potential efficacy of Mn-NiO decorated carbon nanotubes for sensitive determination of glucose.

Keywords: diabetes, glucose, Mn-NiO decorated carbon nanotubes, non-enzymatic

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Authors: M. Merabet-Khelassi, Z. Houiene, L. Aribi-Zouioueche, O. Riant

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Lipases (EC.3.1.1.3) are efficient biotools widely used for their remarkable chemo-, regio- and enantio-selectivity, especially, in kinetic resolution of racemates. They offer access to a large panel of enantiopure building blocks, such as secondary benzylic alcohols, commonly used as synthetic intermediates in pharmaceutical and agrochemical industries. Due to the stability of lipases in both water and organic solvents poor in water, they are able to catalyze both transesterifications of arylalkylcarbinols and hydrolysis of their corresponding acetates. The use of enzymatic hydrolysis in aqueous media still limited. In this presentation, we expose a practical methodology for the preparation of optically enriched acetates using a Candida antarctica lipase B-catalyzed hydrolysis in non-aqueous media in the presence of alkaline carbonate salts. The influence of several parameters which can intervene on the enzymatic efficiency such as the impact of the introduction of the carbonates salts, its amount and the nature of the alkaline earth metal are discussed. The obtained results show that the use of sodium carbonate with CAL-B enhances drastically both reactivity and selectivity of this immobilized lipase. In all cases, the resulting alcohols and remaining acetates are obtained in high ee values (up to > 99 %), and the selectivities reach (E > 500).

Keywords: alkaline-hydrolysis, enzymatic kinetic resolution, lipases, arylalkylcarbinol, non-aqueous media

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Authors: N. Pérez-Rodríguez, D. García-Bernet, A. Torrado-Agrasar, J. M. Cruz, A. B. Moldes, J. M. Domínguez

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World economy is based on non-renewable, fossil fuels such as petroleum and natural gas, which entails its rapid depletion and environmental problems. In EU countries, the objective is that at least 20% of the total energy supplies in 2020 should be derived from renewable resources. Biogas, a product of anaerobic degradation of organic substrates, represents an attractive green alternative for meeting partial energy needs. Nowadays, trend to circular economy model involves efficiently use of residues by its transformation from waste to a new resource. In this sense, characteristics of agricultural residues (that are available in plenty, renewable, as well as eco-friendly) propitiate their valorisation as substrates for biogas production. Corn cob is a by-product obtained from maize processing representing 18 % of total maize mass. Corn cob importance lies in the high production of this cereal (more than 1 x 109 tons in 2014). Due to its lignocellulosic nature, corn cob contains three main polymers: cellulose, hemicellulose and lignin. Crystalline, highly ordered structures of cellulose and lignin hinders microbial attack and subsequent biogas production. For the optimal lignocellulose utilization and to enhance gas production in anaerobic digestion, materials are usually submitted to different pretreatment technologies. In the present work, enzymatic hydrolysis, ultrasounds and combination of both technologies were assayed as pretreatments of corn cob for biogas production. Enzymatic hydrolysis pretreatment was started by adding 0.044 U of Ultraflo® L feruloyl esterase per gram of dry corncob. Hydrolyses were carried out in 50 mM sodium-phosphate buffer pH 6.0 with a solid:liquid proportion of 1:10 (w/v), at 150 rpm, 40 ºC and darkness for 3 hours. Ultrasounds pretreatment was performed subjecting corn cob, in 50 mM sodium-phosphate buffer pH 6.0 with a solid: liquid proportion of 1:10 (w/v), at a power of 750W for 1 minute. In order to observe the effect of the combination of both pretreatments, some samples were initially sonicated and then they were enzymatically hydrolysed. In terms of methane production, anaerobic digestion of the corn cob pretreated by enzymatic hydrolysis was positive achieving 290 L CH4 kg MV-1 (compared with 267 L CH4 kg MV-1 obtained with untreated corn cob). Although the use of ultrasound as the only pretreatment resulted detrimentally (since gas production decreased to 244 L CH4 kg MV-1 after 44 days of anaerobic digestion), its combination with enzymatic hydrolysis was beneficial, reaching the highest value (300.9 L CH4 kg MV-1). Consequently, the combination of both pretreatments improved biogas production from corn cob.

Keywords: biogas, corn cob, enzymatic hydrolysis, ultrasound

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Authors: Miguel Pereira, Lígia Pimentel, Manuela Pintado

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Animal blood is a major by-product of slaughterhouses and still represents a cost and environmental problem in some countries. To be eliminated, blood should be stabilised by cooking and afterwards the slaughterhouses must have to pay for its incineration. In order to reduce the elimination costs and valorise the high protein content the aim of this study was the optimization of hydrolysis conditions, in terms of enzyme ratio and time, in order to obtain hydrolysates with biological activity. Two enzymes were tested in this assay: pepsin and proteases from Cynara cardunculus (cardosins). The latter has the advantage to be largely used in the Portuguese Dairy Industry and has a low price. The screening assays were carried out in a range of time between 0 and 10 h and using a ratio of enzyme/reaction volume between 0 and 5%. The assays were performed at the optimal conditions of pH and temperature for each enzyme: 55 °C at pH 5.2 for cardosins and 37 °C at pH 2.0 for pepsin. After reaction, the hydrolysates were evaluated by FPLC (Fast Protein Liquid Chromatography) and tested for their antioxidant activity by ABTS method. FPLC chromatograms showed different profiles when comparing the enzymatic reactions with the control (no enzyme added). The chromatogram exhibited new peaks with lower MW that were not present in control samples, demonstrating the hydrolysis by both enzymes. Regarding to the antioxidant activity, the best results for both enzymes were obtained using a ratio enzyme/reactional volume of 5% during 5 h of hydrolysis. However, the extension of reaction did not affect significantly the antioxidant activity. This has an industrial relevant aspect in what concerns to the process cost. In conclusion, the enzymatic blood hydrolysis can be a better alternative to the current elimination process allowing to the industry the reuse of an ingredient with biological properties and economic value.

Keywords: antioxidant activity, blood, by-products, enzymatic hydrolysis

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Authors: Payel Ghosh, Rama Chandra Pradhan, Sabyasachi Mishra

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Jamun (Syzygium cuminii L.) is one of the important indigenous minor fruit with high medicinal value. The jamun cultivation is unorganized and there is huge loss of this fruit every year. The perishable nature of the fruit makes its postharvest management further difficult. Due to the strong cell wall structure of pectin-protein bonds and hard seeds, extraction of juice becomes difficult. Enzymatic treatment has been commercially used for improvement of juice quality with high yield. The objective of the study was to optimize the best treatment method for juice extraction. Enzymes (Pectinase and Tannase) from different stains had been used and for each enzyme, best result obtained by using response surface methodology. Optimization had been done on the basis of physicochemical property, nutritional property, sensory quality and cost estimation. According to quality aspect, cost analysis and sensory evaluation, the optimizing enzymatic treatment was obtained by Pectinase from Aspergillus aculeatus strain. The optimum condition for the treatment was 44 ^oC with 80 minute with a concentration of 0.05% (w/w). At these conditions, 75% of yield with turbidity of 32.21NTU, clarity of 74.39%T, polyphenol content of 115.31 mg GAE/g, protein content of 102.43 mg/g have been obtained with a significant difference in overall acceptability.

Keywords: enzymatic treatment, Jamun, optimization, physicochemical property, sensory analysis

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Authors: Leo Paul Vaurs, Sonia Heaven, Charles Banks

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Municipal solid waste (MSW) is a virtually unlimited source of lignocellulosic material in the form of a waste paper/cardboard mixture which can be converted into fermentable sugars via cellulolytic enzyme hydrolysis in a biorefinery. The extraction of the lignocellulosic fraction and its preparation, however, are energy and water demanding processes. The waste water generated is a rich organic liquor with a high Chemical Oxygen Demand that can be partially cleaned while generating biogas in an Upflow Anaerobic Sludge Blanket bioreactor and be further re-used in the process. In this work, an experiment was designed to determine the critical contaminant concentrations in water affecting either anaerobic digestion or enzymatic hydrolysis by simulating multiple water re-circulations. It was found that re-using more than 16.5 times the same water could decrease the hydrolysis yield by up to 65 % and led to a complete granules desegregation. Due to the complexity of the water stream, the contaminant(s) responsible for the performance decrease could not be identified but it was suspected to be caused by sodium, potassium, lipid accumulation for the anaerobic digestion (AD) process and heavy metal build-up for enzymatic hydrolysis. The experimental data were incorporated into a Water Pinch technology based model that was used to optimize the water re-utilization in the modelled system to reduce fresh water requirement and wastewater generation while ensuring all processes performed at optimal level. Multiple scenarios were modelled in which sub-process requirements were evaluated in term of importance, operational costs and impact on the CAPEX. The best compromise between water usage, AD and enzymatic hydrolysis yield was determined for each assumed contaminant degradations by anaerobic granules. Results from the model will be used to build the first MSW based biorefinery in the USA.

Keywords: anaerobic digestion, enzymatic hydrolysis, municipal solid waste, water optimization

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Authors: Delgado-Meza M., Minor-Pérez H.

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Protease is the generic name of enzymes that hydrolyze proteins. These are classified in the subgroup EC3.4.11-99X of the classification enzymes. In food technology the proteolysis is used to modify functional and nutritional properties of food, and in some cases this proteolysis may cause food spoilage. In general, seafood and rainbow trout have accelerated decomposition process once it has done its capture, due to various factors such as the endogenous enzymatic activity that can result in loss of structure, shape and firmness, besides the release of amino acid precursors of biogenic amines. Some studies suggest the use of protease inhibitors from legume as biological regulators of proteolytic activity. The enzyme inhibitors are any substance that reduces the rate of a reaction catalyzed by an enzyme. The objective of this study was to evaluate the reduction of the proteolytic activity of enzymes in extracts of rainbow trout with protease inhibitors obtained from chickpea flour. Different proportions of rainbow trout enzyme extract (75%, 50% and 25%) and extract chickpea enzyme inhibitors were evaluated. Chickpea inhibitors were obtained by mixing 5 g of flour in 30 mL of pH 7.0 phosphate buffer. The sample was centrifuged at 8000 rpm for 10 min. The supernatant was stored at -15°C. Likewise, 20 g of rainbow trout were ground in 20 mL of phosphate buffer solution at pH 7.0 and the mixture was centrifuged at 5000 rpm for 20 min. The supernatant was used for the study. In each treatment was determined the specific enzymatic activity with the technique of Kunitz, using hemoglobin as substrate for the enzymes acid fraction and casein for basic enzymes. Also biuret protein was quantified for each treatment. The results showed for fraction of basic enzymes in the treatments evaluated, that were inhibition of endogenous enzymatic activity. Inhibition values compared to control were 51.05%, 56.59% and 59.29% when the proportions of endogenous enzymes extract rainbow trout were 75%, 50% and 25% and the remaining volume used was extract with inhibitors. Treatments with acid enzymes showed no reduction in enzyme activity. In conclusion chickpea flour reduced the endogenous enzymatic activity of rainbow trout, which may favor its application to increase the half-life of this food. The authors acknowledge the funding provided by the CONACYT for the project 131998.

Keywords: rainbouw trout, enzyme inhibitors, proteolysis, enzyme activity

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Authors: Molin A., De Ramos J. M., Cadion L. G., Pico R. L.

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Malnutrition and food insecurity represent significant global challenges affecting millions of individuals, particularly in low-income and developing regions. The researchers created a solar-powered enzymatic composting bioreactor with an Arduino-based monitoring system for pH, humidity, and temperature. It manages mixed municipal solid wastes incorporating industrial enzymes and whey additives for accelerated composting and minimized carbon footprint. Within 15 days, the bioreactor yielded 54.54% compost compared to 44.85% from traditional methods, increasing yield by nearly 10%. Tests showed that the bioreactor compost had 4.84% NPK, passing metal analysis standards, while the traditional pit compost had 3.86% NPK; both are suitable for agriculture. Statistical analyses, including ANOVA and Tukey's HSD test, revealed significant differences in agricultural yield across different compost types based on leaf length, width, and number of leaves. The study compared the effects of different composts on Brassica rapa subsp. Chinesis (Petchay) and Brassica juncea (Mustasa) plant growth. For Pechay, significant effects of compost type on plant leaf length (F(5,84) = 62.33, η² = 0.79) and leaf width (F(5,84) = 12.35, η² = 0.42) were found. For Mustasa, significant effects of compost type on leaf length (F(4,70) = 20.61, η² = 0.54), leaf width (F(4,70) = 19.24, η² = 0.52), and number of leaves (F(4,70) = 13.17, η² = 0.43) were observed. This study explores the effectiveness of the enzymatic composting bioreactor and its viability in promoting carbon farming as a solution to food insecurity and malnutrition.

Keywords: malnutrition, food insecurity, enzymatic composting bioreactor, arduino-based monitoring system, enzymes, carbon farming, whey additive, NPK level

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Authors: C. C. Castro, T. Senechal, D. Lahem, A. L. Hantson

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Formaldehyde is one of the most representative volatile organic compounds present in the indoor air of residential units and workplaces. Increased attention has been given to this toxic compound because of its carcinogenic effect in health. Biological or enzymatic transformation is being explored to degrade this pollutant. Pseudomonas putida is a bacteria able to synthesize formaldehyde dehydrogenase, an enzyme known to use formaldehyde as a substrate and transform it into less toxic compounds. The immobilization of bacterial cells in the surface of different supports through spraying or dip-coating is herein proposed. The determination of the enzymatic activity on the coated surfaces was performed as well as the study of its effect on formaldehyde degradation in an isolated chamber. Results show that the incorporation of microbial cells able to synthesize depolluting enzymes can be an innovative, low-cost, effective and environmentally friendly solution for indoor air depollution.

Keywords: cells encapsulation, formaldehyde, formaldehyde dehydrogenase, indoor air depollution

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Authors: Onur Ercan, Buket Askin, Erdogan Kucukoner

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Blueberry (Vaccinium corymbosum, bluegold) has become popular beverage due to their nutritional values such as vitamins, minerals, and antioxidants. In the study, the effects of pressing, mashing, enzymatic treatment, and pasteurization on anthocyanins, colour, and polyphenolics of blueberry juice (BJ) were studied. The blueberry juice (BJ) was produced with two different methods that direct juice extraction (DJE) and mash treatment process (MTP) were applied. After crude blueberry juice (CBJ) production, the samples were first treated with commercial enzymes [Novoferm-61 (Novozymes A/S) (2–10 mL/L)], to break down the hydrocolloid polysaccharides, mainly pectin and starch. The enzymes were added at various concentrations. The highest transmittance (%) was obtained for Novoferm-61 at a concentration of 2 mL/L was 66.53%. After enzymatic treatment, clarification trials were applied to the enzymatically treated BJs with adding various amounts of bentonite (10%, w/v), gelatin (1%, w/v) and kiselsol (15%, v/v). The turbidities of the clarified samples were then determined. However, there was no significant differences between transmittances (%) for samples. For that, only enzymatic treatment was applied to the blueberry juice processing (DDBJ, depectinized direct blueberry juice). Based on initial pressing process made to evaluate press function, it was determined that pressing fresh blueberries with no other processing did not render adequate juice due to lack of liquefaction. Therefore, the blueberries were mash into small pieces (3 mm) and then enzymatic treatments and clarification trials were performed. Finally, both BJ samples were pasteurized. Compositional analyses, colour properties, polyphenols and antioxidant properties were compared. Enzymatic treatment caused significant reductions in ACN content (30%) in Direct Blueberry Juice Processing (DBJ), while there was a significant increasing in Mash Treatment Processing (MTP). Overall anthocyanin levels were higher intreated samples after each processing step in MTP samples, but polyphenolic levels were slightly higher for both processes (DBJ and MTP). There was a reduction for ACNs and polyphenolics only after pasteurization. It has a result that the methods for tried to blueberry juice is suitable into obtain fresh juice. In addition, we examined fruit juice during processing stages; anthocyanin, phenolic substance content and antioxidant activity are higher, and yield is higher in fruit juice compared to DBJ method in MTP method, the MTP method should be preferred in processing juice of blueberry into fruit juice.

Keywords: anthocyanins, blueberry, depectinization, polyphenols

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Authors: Selvamuthukumaran Thirunavukkarasu, Arivudainambi Sundararajan

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The plant active molecules with their known mode of action are important leads to the development of newer insecticides. Lactone glycoside was identified earlier as the active principle in Cleistanthus collinus (Roxb.) Benth. (Fam: Euphorbiaceae). It possessed feeding deterrent, insecticidal and insect growth regulatory actions at varying concentrations. Deducing its mode of action opens a possibility of its further development. A no-choice leaf disc bioassay was carried out with lactone glycoside at different doses for different instars and Deterrence Indices were worked out. Using regression analysis concentrations imparting 10, 30 and 50 per cent deterrence (DI10, DI30 & DI50) were worked out. At these doses, effect on nutritional indices like Relative Consumption and Growth Rates (RCR & RGR), Efficiencies of Conversion of Ingested and Digested food (ECI & ECD) and Approximate Digestibility (AD) were worked out. The Relative Consumption and Growth Rate of control and lactone glycoside larva were compared by regression analysis. Regression analysis of deterrence indices revealed that the concentrations needed for imparting 50 per cent deterrence was 60.66, 68.47 and 71.10 ppm for third, fourth and fifth instars respectively. Relative consumption rate (RCR) and relative growth rate (RGR) were reduced. This confirmed the antifeedant action of the fraction. Approximate digestibility (AD) was found greater in treatments indicating reduced faeces because of poor digestibility and retention of food in the gut. Efficiency of conversion of both ingested and digested (ECI and ECD) food was also found to be greatly reduced. This indicated presence of toxic action. This was proved by comparing growth efficiencies of control and lactone glycoside treated larvae. Lactone glycoside was found to possess both feeding deterrent and toxic modes of action. Studies on molecular targets based on this preliminary site of action lead to new insecticide development.

Keywords: Spodoptera litura Fabricius, Cleistanthus collinus (Roxb.) Benth, feeding deterrence, mode of action

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

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

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

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Authors: Rajeev Ravindran, Amit K. Jaiswal

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Waste from the food-processing industry is produced in large amount and contains high levels of lignocellulose. Due to continuous accumulation throughout the year in large quantities, it creates a major environmental problem worldwide. The chemical composition of these wastes (up to 75% of its composition is contributed by polysaccharide) makes it inexpensive raw material for the production of value-added products such as biofuel, bio-solvents, nanocrystalline cellulose and enzymes. In order to use lignocellulose as the raw material for the microbial fermentation, the substrate is subjected to enzymatic treatment, which leads to the release of reducing sugars such as glucose and xylose. However, the inherent properties of lignocellulose such as presence of lignin, pectin, acetyl groups and the presence of crystalline cellulose contribute to recalcitrance. This leads to poor sugar yields upon enzymatic hydrolysis of lignocellulose. A pre-treatment method is generally applied before enzymatic treatment of lignocellulose that essentially removes recalcitrant components in biomass through structural breakdown. Present study is carried out to find out the best pre-treatment method for the maximum liberation of reducing sugars from spent coffee waste (SPW). SPW was subjected to a range of physical, chemical and physico-chemical pre-treatment followed by a sequential, combinatorial pre-treatment strategy is also applied on to attain maximum sugar yield by combining two or more pre-treatments. All the pre-treated samples were analysed for total reducing sugar followed by identification and quantification of individual sugar by HPLC coupled with RI detector. Besides, generation of any inhibitory compounds such furfural, hydroxymethyl furfural (HMF) which can hinder microbial growth and enzyme activity is also monitored. Results showed that ultrasound treatment (31.06 mg/L) proved to be the best pre-treatment method based on total reducing content followed by dilute acid hydrolysis (10.03 mg/L) while galactose was found to be the major monosaccharide present in the pre-treated SPW. Finally, the results obtained from the study were used to design a sequential lignocellulose pre-treatment protocol to decrease the formation of enzyme inhibitors and increase sugar yield on enzymatic hydrolysis by employing cellulase-hemicellulase consortium. Sequential, combinatorial treatment was found better in terms of total reducing yield and low content of the inhibitory compounds formation, which could be due to the fact that this mode of pre-treatment combines several mild treatment methods rather than formulating a single one. It eliminates the need for a detoxification step and potential application in the valorisation of lignocellulosic food waste.

Keywords: lignocellulose, enzymatic hydrolysis, pre-treatment, ultrasound

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Authors: Sri Satya Antarlina, Elok Zubaidah, Teti Istiana, Harijono

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Sorghum bicolor (Sorghum bicolor L. Moench) has the potential as a flour for gluten-free food products. Sorghum flour production needs grain soaking treatment. Soaking can reduce the tannin content which is an anti-nutrient, so it can increase the protein digestibility. Fine particle size decreases the yield of flour, so it is necessary to study various particle sizes to increase the yield. This study aims to determine the characteristics of sorghum flour in the treatment of soaking peeled grain and particle size. The material of white sorghum varieties KD-4 from farmers in East Java, Indonesia. Factorial randomized factorial design (two factors), repeated three times, factor I were the time of grain soaking (five levels) that were 0, 12, 24, 36, and 48 hours, factor II was the size of the starch particles sifted with a fineness level of 40, 60, 80, and 100 mesh. The method of making sorghum flour is grain peeling, soaking peeled grain, drying using the oven at 60ᵒC, milling, and sieving. Physico-chemical analysis of sorghum flour. The results show that there is an interaction between soaking time of grain with the size of sorghum flour particles. Interaction in yield of flour, L* color (brightness level), whiteness index, paste properties, amylose content, protein content, bulk density, and protein digestibility. The method of making sorghum flour through the soaking of peeled grain and the difference in particle size has an important role in producing the physicochemical properties of the specific flour. Based on the characteristics of sorghum flour produced, it is determined the method of making sorghum flour through sorghum grain soaking for 24 hours, the particle size of flour 80 mesh. The sorghum flour with characteristic were 24.88% yield of flour, 88.60 color L* (brightness level), 69.95 whiteness index, 3615 Cp viscosity, 584.10 g/l of bulk density, 24.27% db protein digestibility, 90.02% db starch content, 23.4% db amylose content, 67.45% db amylopectin content, 0.22% db crude fiber content, 0.037% db tannin content, 5.30% db protein content, ash content 0.18% db, carbohydrate content 92.88 % db, and 1.94% db fat content. The sorghum flour is recommended for cookies products.

Keywords: characteristic, sorghum (Sorghum bicolor L. Moench) flour, grain soaking, particle size, physicochemical properties

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Authors: Fana Woldetsadik

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Ethiopia has a huge livestock population, including sheep, that has been contributing a considerable portion to the economy of the country and still promising to rally around the economic advancement of the country. However, feed shortage is a limiting factor in the production and productivity of sheep among Ethiopian smallholder farmers. Therefore, the aim of this study was to prove the role of the locally available brewery by-products called dried Atella as a supplement in feed intake, digestibility, live weight gain, carcass yield, and economic benefit in comparison with commercially purchased supplements known as niger seed cake (NSC). This on-station feeding experiment was conducted on the Zenzelma Campus of Bahir Dar University animal farm. The experimental design used for this research was a completely randomized design (CRD) with five replications. The crude protein (CP) content of dried Atella, wheat bran (WB), natural pasture hay (NPH) and NSC were about 25.07%, 16.57%, 4.48% and 38.04%, respectively, while the neutral detergent fibre (NDF),acid detergent fibre (ADF) and acid detergent lignin (ADL) content of dried Atella, WB, NPH and NSC were around 31.75%, 8.31%, 8.14%; 42.05%, 22.64%, 4.04%; 74.21%, 50.81%, 8.66%; 42.31%, 26.95% and 6.9%, respectively. The result depicted that a higher(P < 0.001) feed intake, nutrient intake, and digestibility for lambs supplemented with Atella than those supplemented with NSC. Furthermore, daily body weight gain and carcass characteristics were better (P < 0.05) for the sheep supplemented with dried Atella than NSC. On the other hand, in terms of profitability, although there was no substantial difference (P > 0.05) between T2 (animals fed NPH,NSC and WB) and T3 (animals fed NPH, Atella and WB), slightly better benefit was recorded in T3 groups. However, loss of money was recorded in T1 (animals fed NPH and WB). Hence, from the biological performance of lambs, it was concluded that Atella could be a potential supplementary feed for sheep fattening among smallholder farmers than NSC despite no profitability difference. Nevertheless, further investigation is recommended to examine the consequence of supplementation of NPH with NSC and NPH with Atella on fatty acid profile analysis, the physicochemical composition of meat, and meat composition.

Keywords: Attela, Bahir Dar university, Carcass yield, digestibility, natural pasture hay, Niger seed cake, smallholder farmers, weight gain, Ethiopia

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Authors: M. S. Kilic, S. Korkut, B. Hazer

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Newly synthesized Polypropylene-g-Polyethylene glycol polymer was first time used for a compartment-less enzymatic fuel cell. Working electrodes based on Polypropylene-g-Polyethylene glycol were operated as unmediated and mediated system (with ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and bilirubin oxidase was selected as anodic and cathodic enzyme, respectively. Glucose was used as fuel in a single-compartment and membrane-less cell. Maximum power density was obtained as 0.65 nW cm-2, 65 nW cm-2, and 23500 nW cm-2 from the unmediated, ferrocene and gold/cobalt oxide modified polymeric film, respectively. Power density was calculated to be ~16000 nW cm-2 for undiluted wastewater sample with gold/cobalt oxide nanoparticles including system.

Keywords: bilirubin oxidase, enzymatic fuel cell, glucose oxidase, nanoparticles

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Authors: Farnaz Alaeimoghadam, Farzad Alaeimoghadam

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In low-lactose milk, due to lactose hydrolysis and its conversion to monosaccharides like glucose and galactose, the Maillard reaction (non-enzymatic browning) occurs more readily compared to non-hydrolyzed milk. This reaction incurs off-flavor and dark color, as well as a decrease in the nutritional value of milk. The target of this research was to evaluate the effect of natural antioxidants in diminishing the browning in low-lactose milk. In this research, three antioxidants, namely ascorbic acid, gallic acid, and pantothenic acid in the concentration range of 0-1 mM/L, either in combination with each other or separately, were added to low-lactose milk. After heat treatment (120 0C for 3 min.), milk samples incubated at 55 0C for one day and then stored at 4 0C for 9 days. Quality indices, including total phenol content, antioxidant activity, color indices, and sensory characters, were measured during intervals of 0, 2, 5, 7, and 9 days. Results of this research showed that the effect of storage time and adding antioxidants were significant on pH, antioxidant activity, total phenolic compounds either before or after heating, index L*, color change, and sensational characteristics (p < 0.05); however, acidity, a* and b* indices, chroma, and hue angle showed no significant changes (p > 0.05). The findings showed that the simultaneous application of gallic acid and ascorbic in the diminishing of non-enzymatic browning and color change, increasing pH, longevity, and antioxidant activity after heat treatment, and augmenting phenolic compounds before heat treatment was better than that of pantothenic acid.

Keywords: Maillard, low-lactose milk, non-enzymatic browning, natural antioxidant

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Authors: Meha Alouini, Wissem Mnif, Yasmine Souissi

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This work will be conducted within the framework of the environmental sustainable development. It involves waste recovering into biodiesel fuel. Low cost feedstocks such as waste of frying oil and animal fats have been utilized to replace refined vegetable oil for biodiesel production. Biodiesel which refers to fatty acid methyl esters (FAME) was carried out by both chemical and enzymatic reaction of transesterification. In order to compare the two studied reactions the obtained biodiesel was characterized by determining its esters content and its fuel properties according to the European standard EN 14214. It was noted that the chemical method gave the product with the best physical property. But the biological one was found more effective for obtaining important ester content. Thus it would be interesting to optimize the enzymatic pathway of production of biodiesel to obtain a better property of biodiesel.

Keywords: biodiesel, fatty acid methyl esters, transesterification, waste frying oil, waste beef fat

Procedia PDF Downloads 476

Authors: Mustafa Ümi̇t Ünal, Nafi̇z Çeli̇ktaş, Aysun Şener, Sara Betül Dolgun, Duygu Keser

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The aim of this study was to compare acid and base pretreatment of switchgrass for bioethanol production. Switchgrass was pretreated with sulfuric acid and sodium hydroxide at 0.5, 1.0 and 1.5% (v/v) at 120, 140, 180 °C for 10, 60 and 90. Optimization of enzymatic hydrolysis of the pretreated switchgrass samples were carried out using three different enzyme mixtures (22.5 mg cellulase and 75 mg cellobiase /g biomass; 45 mg cellulase and 150 mg cellobiase /g biomass; 90 mg cellulase and 300 mg cellobiase /g biomass). Samples were removed at 24-h interval for fermentable sugar analyses with HPLC. The results showed that use of 90 mg cellulase and 300 mg cellobiase/g biomass resulted in the highest fermentable sugar formation. Furthermore, the highest fermentable sugar yield was obtained by pretreatment at 120 °C for 10 min using 1.0 % sodium hydroxide.

Keywords: switchgrass, acid pretreatment, enzymatic hydrolysis, base pretreatment, ethanol production

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Authors: Abinaya Balasubramanian, Satyabrata Ghosh, Satyahari Dey

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Non-Digestible Oligosaccharides(NDOs) are low molecular weight carbohydrates with degree of polymerization (DP) 3-20, that are delivered intact to the large intestine. NDOs are gaining attention as effective prebiotic molecules that facilitate prevention and treatment of several chronic diseases. Recently, NDOs are being obtained by cleaving complex polysaccharides as it results in high yield and also as the former tend to display greater bioactivity. Thelebolus sp. IITKGP BT-12, a recently identified psychrophilic, Ascomycetes fungus has been reported to produce a bioactive extracellular polysaccharide(EPS). The EPS has been proved to possess strong prebiotic activity and anti- proliferative effects. The current study is an attempt to identify and optimise the most suitable method for hydrolysis of the above mentioned novel EPS into NDOs, and further purify and characterise the same. Among physical, chemical and enzymatic methods, enzymatic hydrolysis was identified as the best method and the optimum hydrolysis conditions obtained using response surface methodology were: reaction time of 24h, β-(1,3) endo-glucanase concentration of 0.53U and substrate concentration of 10 mg/ml. The NDOs were purified using gel filtration chromatography and their molecular weights were determined using MALDI-TOF. The major fraction was found to have a DP of 7,8. The monomeric units of the NDOs were confirmed to be glucose using TLC and GCMS-MS analysis. The obtained oligosaccharides proved to be non-digestible when subjected to gastric acidity, salivary and pancreatic amylases and hence could serve as efficient prebiotics.

Keywords: characterisation, enzymatic hydrolysis, non-digestible oligosaccharides, response surface methodology

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Authors: Anna Trusek-Holownia, Andrzej Noworyta

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Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.

Keywords: peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides

Procedia PDF Downloads 414

Authors: N. A. Bamidele, S. O. Obasa, I. O. Taiwo, I. Abdulraheem, O. C. Odebiyi, A. A. Adeoye, O. E. Babalola, O. V. Uzamere

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A feeding trial was conducted to investigate the effect of fermented unripe plantain peel meal (FUP) on growth performance, nutrients digestibility and economic indices of production of Nile tilapia, Oreochromis niloticus fingerlings. Fingerlings (150) of Nile tilapia (1.70±0.1g) were stocked at 10 per plastic tank. Five iso-nitrogenous diets containing 40% crude protein in which maize meal was replaced by fermented unripe plantain peel meal at 0% (FUP0), 25% (FUP25), 50% (FUP50), 75% (FUP75) and 100% (FUP100) were formulated and prepared. The fingerlings were fed at 5% body weight per day for 56 days. There was no significant difference (p > 0.05) in all the growth parameters among the treatments. Feed conversion ratio of 1.35 in fish fed diet FUP25 was not significantly different (P > 0.05) from 1.42 of fish fed diet FUP0. Apparent protein digestibility of 86.94% in fish fed diet FUP100 was significantly higher (p < 0.05) than 70.37% in fish fed diet FUP0 while apparent carbohydrate of 88.34% in fish fed diet FUP0 was significantly different (p < 0.05) from 70.29% of FUP100. Red blood cell (4.30 ml/mm3) of fish fed diet FUP100 was not significantly different from 4.13 ml/mm3 of fish fed diet FUP50. The highest percentage profit of 88.85% in fish fed diet FUP100 was significantly higher than 66.68% in fish fed diet FUP0 while the profit index of 1.89 in fish fed diet FUP100 was significantly different from 1.67 in fish fed diet FUP0. Therefore, fermented unripe plantain peel meal can completely replace maize in the diet of O. niloticus fingerlings.

Keywords: fermentation, fish diets, plantain peel, tilapia

Procedia PDF Downloads 508

Authors: Y. H. Tshela Ntumba, A. Przepiórkowska, M. Prochoń

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Polymeric materials have become an integral part of every aspect of today's industry. They have wide applications, inter alia, in areas such as medicine, food industry and agriculture. In agriculture, for example, they are used for the production of pots, irrigation systems and for soil mulching. The aim of this study was the attempt to produce a biodecomposable agricultural mat, by coating cotton fabric with a blend of carboxylated styrene-butadiene latex (LBSK) containing the enzymatic hydrolyzate of keratin from cattle hair, which would serve as a material for mulching. The production of such material allows the beneficial management of burdensome tannery waste constituted by keratin from cattle hair and at the same time, the production of agricultural mats that much faster undergo decomposition than commonly used polyethylene mats.

Keywords: agricultural mat, biodecomposition, biodegradation, carboxylated butadiene-styrene latex, cellulosic-elastomeric material, keratin hydrolyzate, mulching, protein hydrolyzate

Procedia PDF Downloads 385

Authors: Ajay Kumar Singh, Arvind Kumar, S. P. Singh

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Cellulose nanofibrils (CNF) have shown potential of their extensive use in various fields, including papermaking, due to their unique characteristics. In this study, CNF’s were prepared by fibrillating the pulp obtained from raw materials e.g. bagasse, hardwood and softwood using enzymatic pretreatment followed by mechanical refining. These nanofibrils, when examined under FE-SEM, show that partial fibrillation on fiber surface has resulted in production of nanofibers. Mixing these nanofibers with the unrefined and normally refined fibers show their reinforcing effect. This effect is manifested in observing the improvement in the physical and mechanical properties e.g. tensile index and burst index of paper. Tear index, however, was observed to decrease on blending with nanofibers. The optical properties of paper sheets made from blended fibers showed no significant change in comparison to those made from only mechanically refined pulp. Mixing of normal pulp fibers with nanofibers show increase in ºSR and consequent decrease in drainage rate. These changes observed in mechanical, optical and other physical properties of the paper sheets made from nanofibrils blended pulp have been tried to explain considering the distribution of the nanofibrils alongside microfibrils in the fibrous network. Since usually, paper/boards with higher strength are observed to have diminished optical properties which is a drawback in their quality, the present work has the potential for developing paper/boards having improved strength alongwith undiminished optical properties utilising the concepts of nanoscience and nanotechnology.

Keywords: enzymatic pretreatment, mechanical refining, nanofibrils, paper properties

Procedia PDF Downloads 330

Authors: D. Hephzibah, P. Kumaran, N. M. Saifuddin

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Anaerobic digestion is a well-known technique for sustainable energy recovery from sewage sludge. However, sewage sludge digestion is restricted due to certain factors. Pre-treatment methods have been established in various publications as a promising technique to improve the digestibility of the sewage sludge and to enhance the biogas generated which can be used for energy recovery. In this study, continuous flow microwave (MW) pre-treatment with different intensities were compared by using 5 L semi-continuous digesters at a hydraulic retention time of 27 days. We focused on the effects of MW at different intensities on the sludge solubilization, sludge digestibility, and biogas production of the untreated and MW pre-treated sludge. The MW pre-treatment demonstrated an increase in the ratio of soluble chemical oxygen demand to total chemical oxygen demand (sCOD/tCOD) and volatile fatty acid (VFA) concentration. Besides that, the total volatile solid (TVS) removal efficiency and tCOD removal efficiency also increased during the digestion of the MW pre-treated sewage sludge compared to the untreated sewage sludge. Furthermore, the biogas yield also subsequently increases due to the pre-treatment effect. A higher MW power level and irradiation time generally enhanced the biogas generation which has potential for sustainable energy recovery from sewage treatment plant. However, the net energy balance tabulation shows that the MW pre-treatment leads to negative net energy production.

Keywords: anaerobic digestion, biogas, microwave pre-treatment, sewage sludge

Procedia PDF Downloads 293