Search results for: stabilization of enzymes

Authors: Gabriel S. De Oliveira, Patricia P. Adriani, Christophe Moriseau, Bruce D. Hammock, Felipe S. Chambergo

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Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have high biotechnological interest for the drug design and chemistry transformation for industries. In this study, we describe the identification of substrates and inhibitors of epoxide hydrolase enzyme from the filamentous fungus Trichoderma reesei (TrEH), and these inhibitors showed the fungal growth inhibitory activity. We have used the cloned enzyme and expressed in E. coli to develop the screening in the library of fluorescent substrates with the objective of finding the best substrate to be used in the identification of good inhibitors for the enzyme TrEH. The substrate (3-phenyloxiranyl)-acetic acid cyano-(6-methoxy-naphthalen-2-yl)-methyl ester showed the highest specific activity and was chosen for the next steps of the study. The inhibitors screening was performed in the library with more than three thousand molecules and we could identify the 6 best inhibitors. The IC50 of these molecules were determined in nM and all the best inhibitors have urea or amide in their structure, because It has been recognized that these groups fit well in the hydrolase catalytic pocket of the epoxide hydrolases. Then the growth of T. reesei in PDA medium containing these TrEH inhibitors was tested, and fungal growth inhibition activity was demonstrated with more than 60% of inhibition of fungus growth in the assay with the TrEH inhibitor with the lowest IC50. Understanding how this EH enzyme from T. reesei responds to inhibitors may contribute for the study of fungal metabolism and drug design against pathogenic fungi.

Keywords: epoxide hydrolases, fungal growth inhibition, inhibitor, Trichoderma reesei

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Authors: Abdurzag Kerban, Mostfa M. Abou-Ahmed, Abdelrof M. Ghallab, Mona H. Shaker

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Preservation of semen had a major impact on sheep genetic breeding. The aim of this study was to evaluate the effect of protected fat, probiotic and zinc-enriched diets on semen freezability. Twenty two Barki rams were randomly assigned into four groups; Group I (n=5) was fed the basal diet enriched with 3.7% of dry fat/kg concentration/day, Group II (n=5) was fed a basal diet-enriched with 10gm of probiotic /head/day, Group III (n=6) was fed on the basal diet enriched with 100 ppm of 10% zinc chelated with methionine/kg dry matter/day and Group IV (n=6) was served as control. A pool of three to four ejaculates were pooled from rams within a period of ten weeks. Semen was diluted in egg yolk-Tris diluent and processed in 0.25 ml straw. Motility was evaluated after dilution, before freezing and post-thawing at 0, 1, 2 and 3 hour incubation. Viability index, acrosome integrity and leakage of intracellular enzymes (Aspartat aminotransferase and Alkline phosphatase) were also evaluated. Spermatozoa exhibited highly significant (P<0.01) percentages of motility at 0, 1, 2, and 3 hours incubation after thawing, viability index and acrosome integrity in rams fed a diet enriched with protected fat and zinc groups as compared with probiotic and control groups. Also, the mean value of extracellular leakage of AST was significantly lower in fat and zinc group as compared with probiotic and control groups. In conclusion, semen freezability was improved in animals fed a diet fortified with fat and zinc with no significant improvement in animals fed the probiotic-enriched diet.

Keywords: Barki ram semen, freezing, straw, feed additives

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Authors: Yaara Oppenheimer-Shaanan, Nimrod Nachmias, Marina Campos Rocha, Neta Schlezinger, Noam Dotan, Asaf Levy

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Fusarium oxysporum, a fungus that attacks a broad range of plants and can cause infections in humans, operates across different kingdoms. This pathogen encounters varied conditions, such as temperature, pH, and nutrient availability, in plant and human hosts. The Fusarium oxysporum species complex, pervasive in soils globally, can affect numerous plants, including key crops like tomatoes and bananas. Controlling Fusarium infections can involve biocontrol agents that hinder the growth of harmful strains. Our research developed a computational method to identify toxin domains within a vast number of microbial genomes, leading to the discovery of nine distinct toxins capable of killing bacteria and fungi, including Fusarium. These toxins appear to function as enzymes, causing significant damage to cellular structures, membranes and DNA. We explored biological control using bacteria that produce polymorphic toxins, finding that certain bacteria, non-pathogenic to plants, offer a safe biological alternative for Fusarium management, as they did not harm macrophage cells or C. elegans. Additionally, we elucidated the 3D structures of two toxins with their protective immunity proteins, revealing their function as unique DNases. These potent toxins are likely instrumental in microbial competition within plant ecosystems and could serve as biocontrol agents to mitigate Fusarium wilt and related diseases.

Keywords: microbial toxins, antifungal, Fusarium oxysporum, bacterial-fungal intreactions

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Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

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Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves

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Authors: E. A. Alaswad, H. M. Choudhry, F. Z. Filimban

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Sickle Cell Anemia (SCA) is one type of blood diseases related to autosomal disorder. The sickle shaped red blood cells are the main cause of many problems in the blood vessels and capillaries. Aerva Javanica (J) and Ficus Palmata (P) are medicinal plants that have many popular uses and have been proved their efficacy. The aim of this study was to assess the antioxidants activity and the antisickling effect of J and P extractions. The period of this study, air-dried leaves of J, and P plants were ground and the active components were extracted by maceration in water (W) and methanol (M) as solvents. The antioxidants activity of JW, PW, JM, and PM were assessed by way of the radical scavenging method using 2,2-diphenyl-1-picrylhydrazyl (DPPH). To determine the antisickling effect of J and P extracts. 20 samples were collected from sickle cell anemia patients. Different concentrations of J and P extracts (200 and 110 μg/mL) were added on the sample and incubated. A drop of each sample was examined with light microscope. Normal and sickled RBCs were calculated and expressed as the percent of sickling. The stabilization effect of the extracts was measured by the osmotic fragility test for erythrocytes. The finding suggests as estimated by DPPH method, all the extracts showed an antioxidant activity with a significant inhibition of the DPPH radicals. PM has the least IC50% with 71.49 μg/ml while JM was the most with 408.49 μg/ml. Sickle cells treated with extracts at different concentrations significantly reduced the percentage of sickling compering to control samples. However, JM 200 μg/mL give the highest anti-sickling affect with 17.4% of sickling compared to control 67.5 of sickling while PM at 200 μg/mL showed the highest membrane cell stability. In a conclusion, the results showed that J and P extracts have antisickling effects. Therefore, the Aerva javanica and Ficus palmata may have a role in SCA management and a good impact on the patient's lives.

Keywords: Aerva javanica, antioxidant, antisickling, Ficus palmata, sickle cell anemia

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Authors: Iram Siddique

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Synthetic seed technology is an alternative to traditional micropropagation for production and delivery of cloned plantlets. Synthetic seeds were produced by encapsulating nodal segments of C. angustifolia in calcium alginate gel. 3% (w/v) sodium alginate and 100 mM CaCl2. 2H2O were found most suitable for encapsulation of nodal segments. Synthetic seeds cultured on half strength Murashige and Skoog (MS) medium supplemented with thidiazuron (5.0 µM) + indole -3- acetic acid (1.0 µM) produced maximum number of shoots (10.9 ± 0.78) after 8 weeks of culture exhibiting (78%) in vitro conversion response. Encapsulated nodal segments demonstrated successful regeneration after different period (1-6 weeks) of cold storage at 4 °C. The synthetic seeds stored at 4 °C for a period of 4 weeks resulted in maximum conversion frequency (93%) after 8 weeks when placed back to regeneration medium. The isolated shoots when cultured on half strength MS medium supplemented with 1.0 µM indole -3- butyric acid (IBA), produced healthy roots and plantlets with well developed shoot and roots were successfully hardened off in plastic pots containing sterile soilrite inside the growth chamber and gradually transferred to greenhouse where they grew well with 85% survival rate. Changes in the content of photosynthetic pigments, net photosynthetic rate (PN), superoxide dismutase (SOD) and catalase (CAT) activity in C. angustifolia indicated the adaptation of micropropagated plants to ex vitro conditions.

Keywords: biochemical studies, nodal segments, rooting, synthetic seeds, thidiazuron

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Authors: Juan Antonio Mondragon Sanchez, Ruben Santamaria

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The DNA G-quadruplex is a four-stranded DNA structure formed by stacked planes of four base paired guanines (G-quartet). Guanine rich DNA sequences appear in many sites of genomic DNA and can potential form G-quadruplexes, such as those occurring at 3'-terminus of the human telomeric DNA. The formation and stabilization of a G-quadruplex by small ligands at the telomeric region can inhibit the telomerase activity. In turn, the ligands can be used to down regulate oncogene expression making G-quadruplex an attractive target for anticancer therapy. Many G-quadruplex ligands have been proposed with a planar core to facilitate the pi–pi stacking and electrostatic interactions with the G-quartets. However, many drug candidates are impossibilitated to discriminate a G-quadruplex from a double helix DNA structure. In this context, it is important to investigate the site topology for the interaction of a G-quadruplex with a ligand. In this work, we determine the free energy surface of a G-quadruplex-ligand to study the binding modes of the G-quadruplex (TG4T) with the daunomycin (DM) drug. The complex TG4T-DM is studied using classical molecular dynamics in combination with metadynamics simulations. The metadynamics simulations permit an enhanced sampling of the conformational space with a modest computational cost and obtain free energy surfaces in terms of the collective variables (CV). The free energy surfaces of TG4T-DM exhibit other local minima, indicating the presence of additional binding modes of daunomycin that are not observed in short MD simulations without the metadynamics approach. The results are compared with similar calculations on a different structure (the mutated mu-G4T-DM where the 5' thymines on TG4T-DM have been deleted). The results should be of help to design new G-quadruplex drugs, and understand the differences in the recognition topology sites of the duplex and quadruplex DNA structures in their interaction with ligands.

Keywords: g-quadruplex, cancer, molecular dynamics, metadynamics

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Authors: Musaab Mohammed Ahmed Ali, Vladimir Vodichev

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work deals with an universal control technique or single controller for linear and nonlinear stabilization and tracing control systems. These systems may be structured as SISO and MIMO. Parameters of controlled plants can vary over a wide range. Introduced a novel control systems design method, construction of stable platform orbits using derivative balance, solved transfer function stability preservation problem of linear system under partial substitution of a rational function. Universal controller is proposed as a polar system with the multiple orbits to simplify design procedure, where each orbit represent single order of controller transfer function. Designed controller consist of proportional, integral, derivative terms and multiple feedback and feedforward loops. The controller parameters synthesis method is presented. In generally, controller parameters depend on new polynomial equation where all parameters have a relationship with each other and have fixed values without requirements of retuning. The simulation results show that the proposed universal controller can stabilize infinity number of linear and nonlinear plants and shaping desired previously ordered performance. It has been proven that sensor errors and poor performance will be completely compensated and cannot affect system performance. Disturbances and noises effect on the controller loop will be fully rejected. Technical and economic effect of using proposed controller has been investigated and compared to adaptive, predictive, and robust controllers. The economic analysis shows the advantage of single controller with fixed parameters to drive infinity numbers of plants compared to above mentioned control techniques.

Keywords: derivative balance, fixed parameters, stable platform, universal control

Procedia PDF Downloads 135

Authors: Murali Munisamy, Gauthaman Karunakaran, Mubarak Al-Gahtany, Vivekanandhan Subbiah, M. Manjari Tripati

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Background: Sodium valproate is a widely prescribed broad-spectrum anti-epileptic drug. It shows high inter-individual variability in pharmacokinetics and pharmacodynamics and has a narrow therapeutic range. We evaluated the effects of polymorphic uridine diphosphate glucuronosyltransferase (UGT1A9) metabolizing enzyme on the pharmacokinetics of sodium valproate in the patients with epilepsy who showed toxicity to therapy. Methods: Genotype analysis of the patients was made with polymerase chain–restriction fragment length polymorphism (RFLP) with sequencing. Plasma drug concentrations were measured with reversed phase high-performance liquid chromatography (HPLC) and concentration–time data were analyzed by using a non-compartmental approach. Results: The results of this study suggested a significant genotypic as well as allelic association with valproic acid toxicity for UGT1A9 polymorphic enzymes. The elimination half-life (t 1/2=40.2 h) of valproic acid was longer and the clearance rate (CL=937 ml/h) was lower in the poor metabolizers group of UGT1A9 polymorphism who showed toxicity than in the intermediate metabolizers group (t1/2=35.5 h, CL=1042 ml/h) or the extensive metabolizers group (t1/2=26. h, CL=1,302 ml/h). Conclusion: Our findings suggest that the UGT1A9 genetic polymorphism plays a significant role in the steady state concentration of sodium valproate, and it thereby has an impact on the toxicity of the sodium valproate used in the patients with epilepsy.

Keywords: UGT1A9, sodium valporate, pharmacogenetics, polymorphism

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Authors: Asma Lamin, Anna H. Kaksonen, Ivan Cole, Paul White, Xiao-Bo Chen

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Microbiologically influenced corrosion (MIC) is a process in which microorganisms initiate, facilitate, or accelerate the electrochemical corrosion reactions of metallic components. Several reports documented that MIC accounts for about 20 to 40 % of the total cost of corrosion. Biofilm formation due to the presence of microorganisms on the surface of metal components is known to play a vital role in MIC, which can lead to severe consequences in various environmental and industrial settings. Quorum sensing (QS) system plays a major role in regulating biofilm formation and control the expression of some microbial enzymes. QS is a communication mechanism between microorganisms that involves the regulation of gene expression as a response to the microbial cell density within an environment. This process is employed by both Gram-positive and Gram-negative bacteria to regulate different physiological functions. QS involves production, detection, and responses to signalling chemicals, known as auto-inducers. QS controls specific processes important for the microbial community, such as biofilm formation, virulence factor expression, production of secondary metabolites and stress adaptation mechanisms. The use of QS inhibitors (QSIs) has been proposed as a possible solution to biofilm related challenges in many different applications. Although QSIs have demonstrated some strength in tackling biofouling, QSI-based strategies to control microbially influenced corrosion have not been thoroughly investigated. As such, our research aims to target the QS mechanisms as a strategy for mitigating MIC on metal surfaces in engineered systems.

Keywords: quorum sensing, quorum quenching, biofilm, biocorrosion

Procedia PDF Downloads 90

Authors: Jasna M. Čanadanović-Brunet, Gordana S. Ćetković, Jelena J. Vulić, Sonja M. Djilas, Vesna T. Tumbas Šaponjac, Sladjana M. Stajčić

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Honey serves as a source of natural antioxidants, which are effective in reducing the risk of heart disease, cancer, immune-system decline, cataracts, different inflammatory processes, and also prevent deteriorative oxidation reactions in foods such as enzymatic browning of fruit and vegetables. Honey is a natural saturated sugar solution, but it also contains certain minor constituents, proteins, enzymes, amino and organic acids, lipids, vitamins, phenolic acids, flavonoids and carotenoids. It is consumed in its natural form alone, but also in combination with nuts and various kinds of dried fruits. The aim of this research was to investigate the contribution of dried cherry on phenols (TPh) and flavonoids (Fl) contents and antioxidant activities of honey. Phenolic compounds in Serbian polyfloral (PH), linden (LH) and acacia (AH) honey and also in their mixtures with dried cherry, in 40% mass concentrations (PH40; LH40, AH40), were determined. In comparison to honey, TPh increased 2.25 times for LH40, 2.16 times for AH40 and 1.45 times for PH40, while Fl increased 2.81-fold for PH40, 1.21-fold for LH40 and 1.44-fold for AH40. Antioxidant activity was investigated with two assays, DPPH test and reducing power (RP), and expressed as EC50DPPH and RP0.5 values. The EC50DPPH values were: EC50PH40 = 1.16 mg/ml; EC50LH40= 1.42 mg/ml and EC50AH40= 1.69 mg/ml, while RP0.5 were: RP0.5PH40 = 15.05 mg/ml; RP0.5LH40 = 16.09 mg/ml and P0.5AH40 = 17.60 mg/ml. Our results indicate that supplementation of polyfloral, linden and acacia honey with 40% dried cherry improves antioxidant activity of honey by enriching the phenolic composition.

Keywords: antioxidant activity, dried cherry, honey, phenolics

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Authors: Riya Thomas, Denis Music, Tautgirdas Ruzgas

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The detection of tyrosinase holds considerable interest in the domains of food nutrition and human health due to its significant role in causing a detrimental effect on the colour, flavour, and nutritional value of food as well as in the synthesis of melanin causing skin melanoma. Compared to other conventional analytical techniques, electrochemical (EC) sensors are highly promising owing to their quick response, great sensitivity, ease of use, and low cost. Particularly, titania nanoparticle-based electrochemical sensors have drawn special attention in identifying several biomolecules including enzymes, antibodies, and receptors, owing to their enhanced electrocatalytic activity and electron-accepting properties. In this study, Ti-O film-modified electrode is fabricated using reactive magnetron sputtering, and its affinity towards tyrosinase is examined via electrochemical methods. To comprehend the physiochemical and surface properties-governed electrocatalytic activity of modified electrodes, Ti-O films are grown under various compositional ranges and deposition temperature, and their corresponding electrochemical activity towards tyrosinase is studied. Further, to understand the underlying atomistic mechanisms and electronic-scale electrochemical characteristics, density functional theory (DFT) is employed. The main goal of the current work is to determine the correlation between macroscopic measurements and the underlying atomic properties to improve the tyrosinase activity on Ti-O surfaces. Moreover, this work offers an intriguing new perspective on the use of Ti-O-modified electrodes to detect tyrosinase in the areas of clinical diagnosis, skincare, and food science.

Keywords: density functional theory, electrochemical sensor, Ti-O film, tyrosinase

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Authors: Bhuwan C. Joshi, Atish Prakash, Ajudhia N. Kalia

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Ethnopharmacological relevance: The plant Urtica dioica L. (Urticaceae) is used in various diseases including hepatic ailments. Traditionally, the leaves and roots of the plant are used in jaundice. Objective: The aim of the present work was to evaluate hepatoprotective potential of potent antioxidant from Urtica dioica L. against CCl4 induced hepatotoxicity in-vitro and in-vivo model. Materials and methods: Antioxidant activity of hydro alcoholic extract and its fractions petroleum ether fraction (PEF), ethyl acetate fraction (EAF), n-butanol fraction (NBF) and aqueous fraction (AF) were determined by DPPH radicals scavenging assay. Fractions were subjected to in-vitro cell line study. Further, the most potent fraction (EAF) was subjected to in-vivo study. The in-vivo hepatoprotective active fraction was chromatographed on silica column to isolate the bioactive constituent(s). Structure elucidation was done by using various spectrophotometric techniques like UV, IR, 1H NMR, 13C NMR and MS spectroscopy. Results and conclusion: The ethyl acetate fraction (EAF) of Urtica. dioica L. possessed the potent antioxidant activity viz. DPPH (IC50 78.99 ± 0.17 µg/ml). The in-vitro cell line study showed EAF prevented the cell damage. The EAF significantly attenuated the increased liver enzymes activities in serum and tissue. Column chromatography of most potent antioxidant fraction (EAF) leads to the isolation of 4-hydroxy-3-methoxy cinnamic acid which is responsible for its hepatoprotective potential. Hence, the present study suggests that EAF has significant antioxidant and hepatoprotective potential on CCl4 induced hepatotoxicity in-vitro and in-vivo.

Keywords: Urtica dioica L., antioxidant, HepG2 cell line, hepatoprotective

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Authors: Salina Saddick, Mohamed Afifi, Osama Abuznadah

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This study was carried out to determine the median lethal concentrations (LC50) of Zinc nanoparticles (ZnNPs) on Oreochromis niloticus and Tilapia zillii. The biochemical and molecular potential effects of ZnNPs (500 and 2000 μg L−1) on the antioxidant system in the brain tissue of O. niloticus and T. zillii were investigated. Four hundred fish were used for acute and sub-acute studies. ZnNP LC50 concentrations were investigated in O. niloticus and T. zillii. The effect of 500 and 2000 μg L−1 ZnNPs on brain antioxidants of O. niloticus and T. zillii was investigated. The result indicated that 69 h LC50 was 5.5 ± 0.6 and 5.6 ± 0.4 for O. nilotica and T. zillii, respectively. Fish exposed to 500 μg L−1 ZnNPs showed a significant increase in reduced glutathione (GSH), total glutathione (tGSH) levels, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activity and gene expression. On the contrary, malondialdehyde (MDA) levels significantly decreased. Meanwhile, fish exposed to 2000 μg L−1 ZnNPs showed a significant decrease of GSH, tGSH levels, SOD, CAT, GR, GPx and GST activity and gene expression. On the contrary, MDA levels significantly increased. It was concluded that, the 96 h LC50 of ZnNPs was 5.5 ± 0.6 and 5.6 ± 0.4 for O. nilotica and T. zillii, respectively. ZnNPs in exposure concentrations of 2000 μg/L induced a deleterious effect on the brain antioxidant system of O. nilotica and T. zillii. In contrast, ZnNPs in exposure concentrations of 500 μg L−1 produced an inductive effect on the brain antioxidant system of O. nilotica and T. zillii.

Keywords: ZnNPs, LC50, antioxidants, O. nilotica

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Authors: Ahlem Bahi, Youcef Necib, Sakina Zerizer, Cherif Abdennour, Mohamed Salah Boulakoud

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Mercury (II) is a highly toxic metal which induces oxidative stress in the body. In this study, we aimed to investigate the possible protective effect of olive oil, an antioxidant agent, against experimental mercury toxicity in rat model. Administration of mercuric chloride induced significant increase in serum: ALT, AST, and LPA activities; interleukine1, interleukine6, tumor necrosis factor α (TNFα), creatinine, urea, and uric acid levels. Mercuric chloride also induced oxidative stress, as indicate by decreased tissue of GSH level, GSH-Px, and GST activities along with increase the level of lipid peroxidation. Furthermore, treatment with mercuric chloride caused a marked elevation of kidney and liver weight and decreased body weight. Virgin olive oil treatment markedly reduced elevated serum: AST, ALT, and LPA activities; interleukine1, interleukine6, tumor necrosis factor α (TNFα), creatinine, urea, and uric acid levels and contracted the deterious effects of mercuric chloride on oxidative stress markers changes caused by HgCl2 in tissue as compared to control group. Our results implicate that mercury induced oxidative damage in liver and kidney tissue protected by virgin olive oil, with its antioxidant effects.

Keywords: mercury, antioxidant enzymes, pro-inflammatory cytokine, virgin olive oil, lipid peroxidation

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Authors: Laura Acevedo-Pacheco, Janet Alejandra Gutierrez-Uribe, Lucia Elizabeth Cruz-Suarez, Segio Othon Serna-Saldivar

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Seaweeds can generate changes in the metabolism of lipids; as a consequence, this may diminish cholesterol and other lipids in the blood. However, the consumption of marine algae may also alter the functions of other organs. Therefore, the objective of this research was to study the effect of two different sorts of algae (Ecklonia arborea and Silvetia compressa) in the metabolism of lipids, as well as, in the physiology of the liver. Wistar male rats were fed for two months with independent diets composed of 20% of fat and 2.5% of E. arborea and S. compressa each. Blood parameters (cholesterol, lipoproteins, triglycerides, hepatic enzymes) and triglycerides in the liver were quantified, and also hepatic histology analyses were performed. While S. compressa reduced 18% total cholesterol compared to the positive control, E. arborea increased it 5.8%. Animals fed with S. compressa presented a decrement, compared to the positive control, not only in low density lipoproteins levels (53%) but also in triglycerides (67%). The presence of steatosis in the histologies and the high levels of triglycerides showed an evident lipid accumulation in hepatic tissues of rats fed with both algae. These results indicate that even though S. compressa showed a promising resource to decrease total cholesterol and low-density lipoproteins in blood, a detrimental effect was observed in liver physiology. Further investigations should be made to find out if toxic compounds associated with these seaweeds may cause liver damage especially in terms of heavy metals.

Keywords: brown algae, Eisenia arborea, hepatic metabolism, lipidemic metabolism, Pelvetia compressa, steatosis

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Authors: Samira B. R. Prado, Paulo R. Melfi, Beatriz T. Minguzzi, João P. Fabi

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Fruit softening is the main change that occurs during papaya (Carica papaya L.) ripening. It is characterized by the depolymerization of cell wall polysaccharides, especially the pectic fractions, which causes cell wall disassembling. However, it is uncertain how the modification of the two main pectin polysaccharides fractions (water-soluble – WSF, and oxalate-soluble fractions - OSF) accounts for fruit softening. The aim of this work was to correlate molecular weight changes of WSF and OSF with the gene expression of pectin-solubilizing enzymes (pectinases) during papaya ripening. Papaya fruits obtained from a producer were harvest and storage under specific conditions. The fruits were divided in five groups according to days after harvesting. Cell walls from all groups of papaya pulp were isolated and fractionated (WSF and OSF). Expression profiles of pectinase genes were achieved according to the MIQE guidelines (Minimum Information for publication of Quantitative real-time PCR Experiments). The results showed an increased yield and a decreased molecular weight throughout ripening for WSF and OSF. Gene expression data support that papaya softening is achieved by polygalacturonases (PGs) up-regulation, in which their actions might have been facilitated by the constant action of pectinesterases (PMEs). Moreover, BGAL1 gene was up-regulated during ripening with a simultaneous galactose release, suggesting that galactosidases (GALs) could also account for pulp softening. The data suggest that a solubilization of galacturonans and a depolymerization of cell wall components were caused mainly by the action of PGs and GALs.

Keywords: carica papaya, fruit ripening, galactosidases, plant cell wall, polygalacturonases

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Authors: M. Al Zallouha, Y. Landkocz, J. Brunet, R. Cousin, J. M. Halket, E. Genty, P. J. Martin, A. Verdin, D. Courcot, S. Siffert, P. Shirali, S. Billet

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Toluene is one of the most used Volatile Organic Compounds (VOCs) in the industry. Amongst VOCs, Benzene, Toluene, Ethylbenzene and Xylenes (BTEX) emitted into the atmosphere have a major and direct impact on human health. It is, therefore, necessary to minimize emissions directly at source. Catalytic oxidation is an industrial technique which provides remediation efficiency in the treatment of these organic compounds. However, during operation, the catalysts can release some compounds, called byproducts, more toxic than the original VOCs. The catalytic oxidation of a gas stream containing 1000ppm of toluene on Pd/α-Al2O3 can release a few ppm of benzene, according to the operating temperature of the catalyst. The development of new catalysts must, therefore, include chemical and toxicological validation phases. In this project, A549 human lung cells were exposed in air/liquid interface (Vitrocell®) to gas mixtures derived from the oxidation of toluene with a catalyst of Pd/α-Al2O3. Both exposure concentrations (i.e. 10 and 100% of catalytic emission) resulted in increased gene expression of Xenobiotics Metabolising Enzymes (XME) (CYP2E1 CYP2S1, CYP1A1, CYP1B1, EPHX1, and NQO1). Some of these XMEs are known to be induced by polycyclic organic compounds conventionally not searched during the development of catalysts for VOCs degradation. The increase in gene expression suggests the presence of undetected compounds whose toxicity must be assessed before the adoption of new catalyst. This enhances the relevance of toxicological validation of such systems before scaling-up and marketing.

Keywords: BTEX toxicity, air/liquid interface cell exposure, Vitrocell®, catalytic oxidation

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Authors: Camilo A. Guerrero-Martin, Erik Montes Paez, Marcia C. K. Oliveira, Jonathan Campos, Elizabete F. Lucas

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Asphaltenes precipitation is considered as a formation damage problem, which can reduce the oil recovery factor. It fouls piping and surface installations, as well as cause serious flow assurance complications and decline oil well production. Therefore, researchers have shown an interest in chemical treatments to control this phenomenon. The aim of this paper is to assess the asphaltenes precipitation onset of crude oils in the presence of cardanol, by titrating the crude with n-heptane. Moreover, based on this results obtained at atmosphere pressure, the asphaltenes precipitation onset pressure were calculated to predict asphaltenes precipitation in the reservoir, by using differential liberation and refractive index data of the oils. The influence of cardanol concentrations in the asphaltenes stabilization of three Brazilian crude oils samples (with similar API densities) was studied. Therefore, four formulations of cardanol in toluene were prepared: 0, 3, 5, 10 and 15 m/m%. The formulations were added to the crude at 2:98 ratio. The petroleum samples were characterized by API density, elemental analysis and differential liberation test. The asphaltenes precipitation onset (APO) was determined by titrating with n-heptane and monitoring with near-infrared (NIR). UV-Vis spectroscopy experiments were also done to assess the precipitate asphaltenes content. The asphaltenes precipitation envelopes (APE) were also determined by numerical simulation (Multiflash). In addition, the adequate artificial lift systems (ALS) for the oils were selected. It was based on the downhole well profile and a screening methodology. Finally, the oil flowrates were modelling by NODAL analysis production system in the PIPESIM software. The results of this study show that the asphaltenes precipitation onset of the crude oils were 2.2, 2.3 and 6.0 mL of n-heptane/g of oil. The cardanol was an effective inhibitor of asphaltenes precipitation for the crude oils used in this study, since it displaces the precipitation pressure of the oil to lower values. This indicates that cardanol can increase the oil wells productivity.

Keywords: asphaltenes, NODAL analysis production system, precipitation pressure onset, inhibitory molecule

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Authors: Beenish Saba, Ann D. Christy

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Modern day municipal solid waste landfills are operated and controlled to protect the environment from contaminants during the biological stabilization and degradation of the solid waste. They are equipped with liners, caps, gas and leachate collection systems. Landfill gas is passively or actively collected and can be used as bio fuel after necessary purification, but leachate treatment is the more difficult challenge. Leachate, if not recirculated in a bioreactor landfill system, is typically transported to a local wastewater treatment plant for treatment. These plants are designed for sewage treatment, and often charge additional fees for higher strength wastewaters such as leachate if they accept them at all. Different biological, chemical, physical and integrated techniques can be used to treat the leachate. Treating that leachate with simultaneous power production using microbial fuel cells (MFC) technology has been a recent innovation, reported its application in its earliest starting phase. High chemical oxygen demand (COD), ionic strength and salt concentration are some of the characteristics which make leachate an excellent substrate for power production in MFCs. Different materials of electrodes, microbial communities, carbon co-substrates and temperature conditions are some factors that can be optimized to achieve simultaneous power production and treatment. The advantage of the MFC is its dual functionality but lower power production and high costs are the hurdles in its commercialization and more widespread application. The studies so far suggest that landfill leachate MFCs can produce 1.8 mW/m2 with 79% COD removal, while amendment with food leachate or domestic wastewater can increase performance up to 18W/m3 with 90% COD removal. The columbic efficiency is reported to vary between 2-60%. However efforts towards biofilm optimization, efficient electron transport system studies and use of genetic tools can increase the efficiency of the MFC and can determine its future potential in treating landfill leachate.

Keywords: microbial fuel cell, landfill leachate, power generation, MFC

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Authors: Sayed Mostafa, Mohamed Shebl

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The problem of bread stalling is still one of the most troubling problems for those interested in manufacturing bakery products, as increasing the freshness period of bread is considered one of the most important factors that help encourage this industry due to its important role in reducing expected losses. Therefore, this study aims to improve the quality of pan bread and increase its freshness period by enzymatic treatments, including maltogenic α-amylase (MAA), amyloglucosidase (AGS), glucoseoxidase (GOX) and phospholipase (PhL). Rheological and pasting behavior of wheat flour were estimated in addition to the physical, texture, and sensory parameters of the final product. The addition of MAA resulted in a decrease in peak viscosity, breakdown, setback, and pasting temperature. The addition of MAA also led to a reduction in falling number values. Enzymatic treatments (MAA and PhL) exhibited higher alkaline water retention capacity of pan bread compared to untreated pan bread (control) throughout different storage periods. Furthermore, other enzymes displayed varying effects on bread quality; for instance, AGS enhanced the crust color, while a high concentration of GOX improved the specific volume of the bread. Conclusion: The research findings demonstrate that the enzymatic treatments can significantly improve its quality attributes, such as specific volume, increase the alkaline water retention capacity with lower hardness value, which reflects bread freshness during storage periods, and improve sensory characteristics.

Keywords: anti-stalling agents, enzymatic treatments, maltogenic α-amylase, amyloglucosidase, glucoseoxidase, phospholipase, pasting behavior, wheat flour

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Authors: Hala Demerdash, Ola M. Zanaty, Emad Eldin Arida

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Background: This study investigated the micoRNA expression changes induced by Sevoflurane and Propofol and their effects on liver functions. Patients and methods: The study was designed as randomized controlled study, carried out on 200 adult patients, scheduled for major surgeries under general anesthesia (GA). Patients were randomly divided into four groups; groups SC and PC included chronic hepatitis C (CHC) patients where SC group are patients receiving Sevoflurane, and PC group are patients receiving Propofol anesthesia. While S and P groups included non- hepatitis patients; S group are patients receiving Sevoflurane and P group are patients receiving Propofol. Anesthesia in Group S and SC patients was maintained by sevoflurane, while anesthesia in Group P and PC patients was maintained by propofol infusion. Blood samples were analyzed for PT, PTT and liver enzymes. Serum samples were analyzed for microRNA before and after surgery. Results: Results show miRNA-122 and miRNA-21 were absent in serum of S and P groups in pre-operative samples. However, they were expressed in SC and PC groups. In post-operative samples; miRNA-122 revealed an increased expression in all groups; with more exaggerated response in SC group. On the other hand miRNA-21 revealed increased expression in both SC and PC groups; a slight expression in S group with absent expression in P group. There was a post-operative negative correlation between miR-122 and ALT (r=-0.46) in SC group and (r=-0.411) in PC group and positive correlation between ALT and miR-21 (r=0.335) in SC group and (r=0.379) in PC group. The amount of blood loss was positively correlated with miR-122 (r=0.366) in SC group and (r=0.384) in PC group. Conclusion: Propofol anesthesia is safer than Sevoflurane anesthesia in patients with CHC. Sevoflurane and Propofol anesthesia affect miRNA expression in both CHC and non-hepatitis patients.

Keywords: anesthesia, chronic hepatitis C, micoRNA, propofol, sevoflurane

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Authors: Pornrut Rabintossaporn, Suphaket Saenthaweesuk, Amornnat Thuppia, Nuntiya Somparn

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Several dietary and herbal plants have been shown to possess cytoprotective and antioxidant effects with various mechanisms of action. The aim of this study was to determine the antioxidant effects and its mechanism of aqueous leaves extract of Ocimum americanum (OA), commonly known as American basil or 'hoary basil', in rat. The extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with the extract at the dose of 100, 200 and 400 mg/kg for 28 days. Phytochemical screening of plant extracts revealed the presence of alkaloid, cardiac glycosides, tannin and steroid compounds. The extract contained phenolic compounds 36.91 ± 0.66 mg of gallic acid equivalents per gram OA extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 41.27 ± 1.86 µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14µg/mL. In the animals, the extract was well tolerated by the animals throughout the 28 days of study as shown by normal serum levels AST, ALP, ALT, BUN and Cr as well as normal histology of liver and pancreatic and kidney tissue. The protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased compared with normal control. Consistent with the induction of γ-GCL protein expression significantly reduction of serum oxidative stress marker malondialdehyde (MDA) was found in rat treated with OA extract compared with control. Taken together, this study provides evidence that Ocimum americanum exhibits direct antioxidant properties and can induce cytoprotective enzyme in vivo.

Keywords: antioxidant, γ-glutamylcysteine ligase, MDA, Ocimum americanum

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Authors: Nenny Harijani, Dhandy Koesoemo Wardhana

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The aim of this study is to know the bacteriocin as antimicrobial activity produced by Lactic Acid Bacteria (LAB) as Antibacterial of Sub Clinic Mastitis on Dairy Cows. The antimicrobial is produced by LAB which isolates from cattle intestine can inhibit the growth Staphylococcus aureus, Steptocococcus agalactiae an Escherichia coli which were caused by dairy cattle subclinical mastitis. The failure of this bacteria growth was indicated by the formation of a clear zone surrounding the colonies on Brain Heart Infusion Agar plate. The bacteriocin was produced by Lactic Acid Bacteria (LAB) as antimicrobial, which could inhibit the growth of indicator bacteria Staphylococcus aureus, S.aglactiae and E.coli. This study was also developed bacteriocin to be used as a therapeutic of subclinical mastitis on dairy cows. The method used in this study was isolation, selection and identification of LAB using Mann Rogosa Sharp Medium, followed by characterization of the bacteriocin produced by LAB. The result of the study showed that bacteriocin isolated from beef cattle’s intestine could inhibit the growth Staphylococcus aureus, S. agalactiae, an Escherichia coli, which was indicated by clear zone surrounding the colonies on Brain Heart Infusion Agar plate. Characteristics of bacteriocin were heat-stable exposed to 80 0C for 30 minutes and 100 ⁰C for 15 minutes and inactivated by proteolytic enzymes such as trypsin. This approach has suggested the development of bacteriocin as a therapeutic agent for subclinical mastitis in dairy cattle.

Keywords: lactic acid bacteria, bacteriocin, staphylococcus aureus, S. agalactiae, E. coli, sub

Procedia PDF Downloads 134

Authors: Benchouala Amira, Bojic Clement, Poupin Pascal, Cossu Leguille-carole

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The objective of this study is to evaluate in vitro the cytotoxic and androgenic potential of several antifungal molecules (amphotericin B, econazole, ketoconazole and miconazole) on MDA-Kb2 cell lines. This biological model is an effective tool for the detection of endocrine disruptors because it responds well to the main agonist of the androgen receptor (testosterone) and also to an antagonist: flutamide. The cytotoxicity of each chemical compound tested was measured using an MTT assay (tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) which measures the activity of the reductase function of mitochondrial succinate dehydrogenase enzymes of cultured cells. This complementary cytotoxicity test is essential to ensure that the effects of reduction in luminescence intensity observed during androgenic tests are only attributable to the anti-androgenic action of the compounds tested and not to their possible cytotoxic properties. Tests of the androgenic activity of antifungals show that these compounds do not have the capacity to induce transcription of the luciferase gene. These compounds do not exert an androgenic effect on MDA-Kb2 cells in culture for the environmental concentrations tested. The addition of flutamide for the same tested concentrations of antifungal molecules reduces the luminescence induced by amphotericin B, econazole and miconazole, which is explained by a strong interaction of these molecules with flutamide which may have a greater toxic effect than when tested alone. The cytotoxicity test shows that econazole and ketoconazole can cause cell death at certain concentrations tested. This cell mortality is perhaps induced by a direct or indirect action on deoxyribonucleic acid (DNA), ribonucleic acid (RNA) or proteins necessary for cell division.

Keywords: cytotoxicity, androgenic potential, antifungals, MDA-Kb2

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Authors: Sohair Hassan, Olfat Hammam, Sahar Hussein, Wessam Magdi

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Objective: Acute liver failure (ALF) is a clinical condition with an unclear history of pathophysiology, making it a challenging task for scientists to reverse the disease in its initial phase and to help the liver re-function customary: this study aimed to estimate the hepatoprotective effects of Punica granatum Lpeel and Pistacia atlantica leaves as a multi-rich antioxidants ingredients either in their normal and/or in their nanoforms against thioacetamide induced acute liver failure in a rodent model. Method: Male Wistar rats (n=60) were divided into six equal groups, the first group employed as a control; The second group administered a dose of 350 mg /Kg/ b.w of thioacetamide (TAA)-IP, from the third to the sixth group received TAA + [2mls / 100 g b.w/d] of aqueous extracts of Punica granatum L and Pistacia atlantica either in their normal and/or Nano forms consecutively for (14 days) Results: Recorded significant elevation in liver enzymes, lipid profiles, LPO (p= 0.05) and NO with a marked significant decrease in GSH and SOD accompanied by an elevation in inflammatory cytokine (IL6, TNF-α, and AFP) in addition to a noticeable increase in HSP70 level & degradation in DNA respectively in TAA challenged group. However significant and subsequent amelioration of most of the impaired markers was observed with ip nano treatment of both extracts. Conclusion: The current results highlighted the high performance of both plant nano extracts and their hepatoprotective impact and their possible therapeutic role in the amelioration of TAA induced acute liver failure in experimental animals.

Keywords: acute liver failure HPLC, IL6, nano extracts, thioacetamide, TNF-α

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Authors: Mathias Twizeyimana, Urmila Adhikari, Julius P. Sserumaga, David Ingham

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The management of aflatoxins (naturally occurring toxins produced by certain fungi, most importantly Aspergillus flavus and A. parasiticus) relies mostly on the use of best cultural practices and, in some cases, the use of the biological control consisting of atoxigenic strains inhibiting the toxigenic strains through competition resulting in considerable toxin reduction. At AgBiome, we have built a core collection of over 100,000 fully sequenced microbes from diverse environments and employ both the microbes and their sequences in the discovery of new biological products for disease and pest control. The most common approach to finding beneficial microbes consists of isolating microorganisms from samples collected from diverse environments, selecting antagonistic strains through empirical screening, studying modes of action, and stabilization through the formulation of selected microbial isolates. A total of 608 diverse bacterial strains were screened using a high-throughput assay (48-well assay) to identify strains that inhibit toxigenic A. flavus growth on maize kernels. Active strains in 48-well assay had their pathogen inhibiting activity confirmed using the Flask Assay and were concurrently tested for their ability to reduce the aflatoxin content in maize grains. Strains with best growth inhibition and reduction of aflatoxin were tested in the greenhouse and field trials. From the field trials, three bacterial strains, AFS000009 (Pseudomonas chlororaphis), AFS032321 (Bacillus subtilis), AFS024683 (Bacillus velezensis), had aflatoxin concentrations (ppb) values that were significantly lower than those of inoculated control. The identification of biological products with high efficacy in inhibiting pathogen growth and eventually reducing the aflatoxin content will provide a valuable alternative to control strategies used in aflatoxin contamination management.

Keywords: aflatoxin, microorganism bacteria, biocontrol, beneficial microbes

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Authors: Konstans Ruseva, Kristina Ivanova, Katerina Todorova, Margarita Gabrashanska, Tzanko Tzanov, Elena Vassileva

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Zwitterionic polymers (ZP) are well-known with their ultralow biofouling. They are successfully competing with poly(ethylene glycols) (PEG), which are considered as the “golden standard” in this respect. These unique properties are attributed to their strong hydration capacity, defined by the dipole-dipole interactions, arising between the ZP pendant groups as well as to the dipoles interaction with water molecules. Beside, ZP are highly resistant to bacterial adhesion thus ensuring an excellent anti-biofilm formation ability. Moreover, ZP are able to respond upon external stimuli such as temperature, pH, salt concentration changes which in combination with their anti-biofouling effect render this type of polymers as materials with a high potential in biomedical applications. The present work is focused on the development of zwitterionic hydrogels for efficient treatment of highly exudating and hard-to-heal chronic wounds. To this purpose, two types of ZP networks with different crosslinking degree were synthesized - polysulfobetaine (PSB) and polycarboxybetaine (PCB) ones. They were characterized in terms of their physico-mechanical properties, e.g. microhardness, swelling ability, smart behaviour. Furthermore, the potential of ZP networks to resist biofilm formation towards Staphylococcus aureus and Escherichia coli was studied. Their ability to reduce the high levels of myeloperoxidase and metalloproteinase, two enzymes that are part of the chronic wounds enviroenment, was revealed. Moreover, the in vitro cytotoxic assessment of PSB and PCB networks along with their in vivo performance in rats was also studied to reveal their high biocompatibility.

Keywords: absorption properties, biocompatibility, enzymatic inhibition activity, wound healing, zwitterionic polymers

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Authors: Salama A. Ouf, Amera A. El-Adly, Abdelaleam H. Mohamed

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Dermatophytosis is the infection of keratinized tissues such as hair, nail and the stratum corneum of the skin by dermatophytic fungi. Infection is generally cutaneous and restricted to the non-living cornified layers because of the inability of the fungi to penetrate the deeper tissues or organs of immunocompetent hosts. In Saudi Arabia, Onychomycosis is the most frequent infection (40.3%), followed by tinea capitis (21.9%), tinea pedis (16%), tinea cruris (15.1%), and tinea corporis (6.7%). Several azole compounds have been tried to control dermatophytic infection, however, the azole-containing medicines may interfere with the activity of hepatic microsomal enzymes, sex and thyroid hormones, and testosterone biosynthesis. In this research, antibody-conjugated nanoparticles stimulated by cold plasma and laser were evaluated in vitro against some dermatophytes isolated from the common types of tinea. Different types of nanomaterials were tested but silver nanoparticles (AgNPs) were proved to be most effective against the dermatophytes under test. The use of cold plasma coupled with antibody-conjugated nano-particles has severe impact on dermatophytes where the inhibition of growth, spore germination keratinase activity was more than 88% in the case of Trichophyton rubrum, T. violaceum, Microsprum canis and M. gypseum. Complete inhibition of growth for all dermatophytes was brought about by the interaction of conjugated nanoparticles, with cold plasma and laser treatment. The in vivo test with inoculated guinea pigs achieved promising results where the recovery from the infection reached 95% in the case of M. canis –inoculated pigs treated with AgNPs pretreated with cold plasma and laser.

Keywords: cold plasma, dermatophytes, laser, silver nanoparticles

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Authors: Oluwatosin Adetola Arojojoye, Olajumoke Olufunlayo Alao, Philip Odigili

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This study investigated the extent of pollution in Eleyele River in Oyo State, Nigeria by investigating the antioxidant status and malondialdehyde levels (index of lipid peroxidation) in the organs of African Catfish, Clarias gariepinus from the river. Clarias gariepinus weighing between 250g-400g were collected from Eleyele River (a suspected polluted river) and Clarias gariepinus from a clean fish farm (Durantee fisheries) were used as the control. Levels of malondialdehyde, glutathione concentration (GSH) and activities of antioxidant enzymes - superoxide dismutase, catalase and glutathione-S-transferase (GST) were evaluated in the post-mitochondrial fractions of the liver, kidney and gills of the fishes. From the results, there were increases in malondialdehyde level and GSH concentration in the liver, kidney and gills of Clarias gariepinus from Eleyele River when compared with control. Glutathione-S-transferase activity was induced in the liver and kidney of Clarias gariepinus from Eleyele River when compared with control. However, the activity of this enzyme was depleted in the gills of fishes from Eleyele River compared with control. Also there was an induction in SOD activity in the liver of Clarias gariepinus from Eleyele River when compared with control but there was a decrease in the activity of this enzyme in the kidney and gills of fishes from Eleyele River compared with control. Increase in lipid peroxidation and alterations in antioxidant system in Clarias gariepinus from Eleyele River show that the fishes were under oxidative stress. These suggest that the river is polluted probably as a result of industrial, domestic and agricultural wastes frequently discharged into the river. This could pose serious health risks to consumers of water and aquatic organisms from the river.

Keywords: antioxidant, lipid peroxidation, Clarias gariepinus, Eleyele River

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