World Academy of Science, Engineering and Technology

Authors: A. A. M. Azoddein, R. M. Yunus, N. M. Sulaiman, A. B. Bustary, K. Sabar

Abstract:

Microbes have been used to solve environmental problems for many years. The role of microorganism to sequester, precipitate or alter the oxidation state of various heavy metals has been extensively studied. Treatment using microorganism interacts with toxic metal are very diverse. The purpose of this research is to remove the mercury using Pseudomonas putida (P. putida), pure culture ATTC 49128 at optimum growth parameters such as techniques of culture, acclimatization time and speed of incubator shaker. Thus, in this study, the optimum growth parameters of P. putida were obtained to achieve the maximum of mercury removal. Based on the optimum parameters of P. putida for specific growth rate, the removal of two different mercury concentration, 1 ppm and 4 ppm were studied. From mercury nitrate solution, a mercuryresistant bacterial strain which is able to reduce from ionic mercury to metallic mercury was used to reduce ionic mercury. The overall levels of mercury removal in this study were between 80% and 89%. The information obtained in this study is of fundamental for understanding of the survival of P. putida ATTC 49128 in mercury solution. Thus, microbial mercury removal is a potential bioremediation for wastewater especially in petrochemical industries in Malaysia.

Keywords: Pseudomonas putida, growth kinetic, biosorption, mercury, petrochemical wastewater.

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Authors: Wen-Shyong Tzou, Chih-Ching Huang, Chin-Hwa Hu, Ying-Tsang Lo, Tun-Wen Pai, Chia-Yin Chiang, Chung-Hao Li, Hong-Jyuan Jian

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Plasmin plays an important role in the human circulatory system owing to its catalytic ability of fibrinolysis. The immediate injection of plasmin in patients of strokes has intrigued many scientists to design vectors that can transport plasmin to the desired location in human body. Here we predict the structure of human plasmin and investigate the interaction of plasmin with the gold-nanoparticle. Because the crystal structure of plasminogen has been solved, we deleted N-terminal domain (Pan-apple domain) of plasminogen and generate a mimic of the active form of this enzyme (plasmin). We conducted a simulated annealing process on plasmin and discovered a very large conformation occurs. Kringle domains 1, 4 and 5 had been observed to leave its original location relative to the main body of the enzyme and the original doughnut shape of this enzyme has been transformed to a V-shaped by opening its two arms. This observation of conformational change is consistent with the experimental results of neutron scattering and centrifugation. We subsequently docked the plasmin on the simulated gold surface to predict their interaction. The V-shaped plasmin could utilize its Kringle domain and catalytic domain to contact the gold surface. Our findings not only reveal the flexibility of plasmin structure but also provide a guide for the design of a plasmin-gold nanoparticle.

Keywords: Docking, gold nanoparticle, molecular simulation, plasmin.

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Authors: Hyun June Kim, Tailong Shi, Seden Akdagli, Sam Most, Yuling Yan

Abstract:

Quantitative analyses of whisker movements provide a means to study functional recovery and regeneration of mouse facial nerve after an injury. However, accurate tracking of the mouse whisker movement is challenging. Most methods for whisker tracking require manual intervention, e.g. fixing the head of the mouse during a study. Here we describe a semi-automated image processing method, which is applied to high-speed video recordings of free-moving mice to track the whisker movements. We first track the head movement of a mouse by delineating the lower head contour frame-by-frame that allows for detection of the location and orientation of the head. Then, a region of interest is identified for each frame; the subsequent application of a mask and the Hough transform detects the selected whiskers on each side of the head. Our approach is used to examine the functional recovery of damaged facial nerves in mice over a course of 21 days.

Keywords: Mystacial macrovibrissae, whisker tracking, head tracking, facial nerve recovery.

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Authors: Justina I. R. Udotong

Abstract:

Toxicity of copper (Cu), lead (Pb) and iron (Fe) to Tilapia guinensis was carried out for 4 days with a view to determining their effects on the liver and muscle tissues. Tilapia guinensis samples of about 10 - 14cm length and 0.2 – 0.4kg weight each were obtained from University of Calabar fish ponds and acclimated for three (3) days before the experimental set up. Survivors after the 96-hr LC50 test period were selected from test solutions of the heavy metals for the histopathological studies. Histological preparations of liver and muscle tissues were randomly examined for histopathological lesions. Results of the histological examinations showed gross abnormalities in the liver tissues due to pathological and degenerative changes compared to liver and muscle tissues from control samples (tilapia fishes from aquaria without heavy metals). Extensive hepatocyte necrosis with chronic inflammatory changes was observed in the liver of fishes exposed to Cu solution. Similar but less damaging effects were observed in the liver of fishes exposed to Pb and Fe. The extent of lesion observed was therefore heavy metal-related. However, no pathologic changes occurred in the muscle tissues.

Keywords: Degenerative changes, heavy metal, hepatocyte necrosis, histopathology, toxicity.

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Authors: Ibrahim M. S. Shnawa, Bashar A. H. E. Alsadi, Kalida K. Alniaem

Abstract:

In the immunologic sense, clinical infection is a state of failure of the immune system to combat the pathogenic weapon of the bacteria invading the host. A motile gram negative vibroid organism associated with marked mono and poly nuclear cell responses was traced during the examination of a clinical material from an infected common carp Cyprinus carpio. On primary plate culture, growth was shown to be pure, dense population of an Aeromonas-like colony morphotype. The pure isolate was found to be; Aerobic, facultatively anaerobic, non-halophilic, grew at 0C, and 37C, oxidase positive utilizes glucose through fermentative pathway, resist 0/129 and novobiocin, produces alanine and lysine decarboxylases but non-producing ornithine dehydrolases. Tests for the in vitro determinants of pathogenicity has shown to be; Betahaemolytic onto blood agar, gelatinase, casienase and amylase producer. Three in vivo determinants of pathogenicity were tested as, the lethal dose fifty, the pathogenesis and pathogenicity. It was evident that 0.1 milliliter of the causal bacterial cell suspension of a density 1 x 107 CFU/ml injected intramuscularly into an average of 100gms fish toke five days incubation period, then at the day six morbidity and mortality were initiated. LD50 was recorded at the day 12 post-infection. Use of an LD50 doses to study the pathogenicity, reveals mononuclear and polynuclear cell responses, on examining the stained direct films of the clinical materials from the experimentally infected fish. Re-isolation tests confirm that the reisolant is same. The course of the infection in natural case was shown manifestation of; skin ulceration, haemorrhage and descaling. On evisceration, the internal organs were shown; congestion in the intestines, spleen and, air sacs. The induced infection showed a milder form of these manifestations. The grading of the virulence of this organism was virulent causing chronic course of infections as indicated from the pathogenesis and pathogenicity studies. Thus the infectious bacteria were consistent with Aeromonas hydrophila, and the infection was chronic.

Keywords: Piscan, inflammatory respnonse, pure culture, pathogen, chronic, infection.

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Authors: S. Shalaweh, P. Bouic, F. Weitz, R. Henkel

Abstract:

More than 3000 plants of notable phyto-therapeutic value grow in South Africa; these include Cissampelos capensis, commonly known in Afrikaans as dawidjie or dawidjiewortel. C. capensis is the most significant and popular medicinal plant used by the Khoisan as well as other rural groups in the Western region of South Africa. Its rhizomes are traditionally used to treat male fertility problems. Yet, no studies have investigated the effects of this plant or its extracts on human spermatozoa. Therefore, this study aimed at investigating the effects of C. capensis rhizome extract (CRE) fractions on ejaculated human spermatozoa in vitro. Spermatozoa from a total of 77 semen samples were washed with human tubular fluid medium supplemented with bovine serum albumin (HTF-BSA) and incubated for 2 hours with 20 μg/ml progesterone (P4) followed by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200 μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH, F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A sample without addition of CRE fractions served as control. Samples were analyzed for sperm motility, reactive oxygen species (ROS), DNA-fragmentation, acrosome reaction and capacitation. Results showed that F1 resulted in significantly higher values for ROS, capacitation and hyper-activation compared to F2, F3, and F4 with P4-stimulated samples generally having higher values. No significant effect was found for the other parameters. In conclusion, alkaloids present in F1 of CRE appear to have triggered sperm intrinsic ROS production leading to sperm capacitation and acrosome reaction induced by P4.

Keywords: Capacitation, acrosome reaction, Cissampelos capensis, DNA fragmentation, ROS.

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Authors: Nichole Haag, Kimberly Velk, Tyler McCune, Chun Wu

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Methicillin/multiple-resistant Staphylococcus aureus (MRSA) are infectious bacteria that are resistant to common antibiotics. A previous in silico study in our group has identified a hypothetical protein SAV1226 as one of the potential drug targets. In this study, we reported the bioinformatics characterization, as well as cloning, expression, purification and kinetic assays of hypothetical protein SAV1226 from methicillin/vancomycin-resistant Staphylococcus aureus Mu50 strain. MALDI-TOF/MS analysis revealed a low degree of structural similarity with known proteins. Kinetic assays demonstrated that hypothetical protein SAV1226 is neither a domain of an ATP dependent dihydroxyacetone kinase nor of a phosphotransferase system (PTS) dihydroxyacetone kinase, suggesting that the function of hypothetical protein SAV1226 might be misannotated on public databases such as UniProt and InterProScan 5.

Keywords: Dihydroxyacetone kinase, essential genes, Methicillin-resistant Staphylococcus aureus, drug target.

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Authors: Sunil Dehipawala, Aregama Sirisumana, P. Schneider, G. Tremberger Jr, D. Lieberman, Todd Holden T. Cheung

Abstract:

The arsenic and iron environments in different growth stages have been studied with EXAFS and XANES using Brookhaven Synchrotron Light Source. Collard Greens plants were grown and tissue samples were harvested. The project studied the EXAFS and XANES of tissue samples using As and Fe K-edges. The Fe absorption and the Fourier transform bond length information were used as a control comparison. The Fourier transform of the XAFS data revealed the coexistence of As (III) and As (V) in the As bonding environment inside the studied plant tissue samples, although the soil only had As (III). The data suggests that Collard Greens has a novel pathway to handle arsenic absorption in soil.

Keywords: EXAFS, Fourier Transform, metalloproteins, XANES.

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Authors: Sunil Dehipawala, Gayathrie Amarasuriya, N. Gadura, G. Tremberger Jr, D. Lieberman, Harry Gafney, Todd Holden, T. Cheung

Abstract:

The iron environment in Fe-doped Vycor Anode was investigated with EXAFS using Brookhaven Synchrotron Light Source. The iron-reducing Shewanella oneidensis culture was grown in a microbial fuel cell under anaerobic respiration. The Fe bond length was found to decrease and correlate with the amount of biofilm growth on the Fe-doped Vycor Anode. The data suggests that Fe-doped Vycor Anode would be a good substrate to study the Shewanella oneidensis nanowire structure using EXAFS.

Keywords: EXAFS, Fourier Transform, Microbial Fuel Cell, Shewanella oneidensis.

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Authors: S. C. Santos, S. R. Dionísio, A. L. D. De Andrade, L. R. Roque, A. C. Da Costa, J. L. Ienczak

Abstract:

In this work, two fermentations at different temperatures (25 and 30ºC), with cell recycling, were accomplished to produce ethanol, using a mix of commercial substrates, xylose (70%) and glucose (30%), as organic source for Scheffersomyces stipitis. Five consecutive fermentations of 80 g L-1 (1º, 2º and 3º recycles), 96 g L-1 (4º recycle) and 120 g L-1 (5º recycle)reduced sugars led to a final maximum ethanol concentration of 17.2 and 34.5 g L-1, at 25 and 30ºC, respectively. Glucose was the preferred substrate; moreover xylose startup degradation was initiated after a remaining glucose presence in the medium. Results showed that yeast acid treatment, performed before each cycle, provided improvements on cell viability, accompanied by ethanol productivity of 2.16 g L-1 h- 1 at 30ºC. A maximum 36% of xylose was retained in the fermentation medium and after five-cycle fermentation an ethanol yield of 0.43 g ethanol/g sugars was observed. S. stipitis fermentation capacity and tolerance showed better results at 30ºC with 83.4% of theoretical yield referenced on initial biomass.

Keywords: 5-carbon sugar, cell recycling fermenter, mixed sugars, xylose-fermenting yeast.

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Authors: Ahmad Ali Shahid, Muhammad Shakil Shaukat, Kamran Shehzad Bajwa, Abdul Qayyum Rao, Tayyab Husnain

Abstract:

Agriculture is the backbone of economy of Pakistan and cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat severe problems of insect and weed, combination of three genes namely Cry1Ac, Cry2A and EPSPS genes was transferred in locally cultivated cotton variety MNH-786 with the use of Agrobacterium mediated genetic transformation. The present study focused on the molecular screening of transgenic cotton plants at T3 generation in order to confirm integration and expression of all three genes (Cry1Ac, Cry2A and EPSP synthase) into the cotton genome. Initially, glyphosate spray assay was used for screening of transgenic cotton plants containing EPSP synthase gene at T3 generation. Transgenic cotton plants which were healthy and showed no damage on leaves were selected after 07 days of spray. For molecular analysis of transgenic cotton plants in the laboratory, the genomic DNA of these transgenic cotton plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty (Cry1Ac gene), ten out of twenty (Cry2A gene) and all twenty (EPSP synthase gene) were produced positive amplification. On the base of PCR amplification, ten transgenic plant samples were subjected to protein expression analysis through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the mRNA expression levels of Cry1Ac and EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes at T3 generation.

Keywords: Agriculture, Cotton, Transformation, Cry Genes, ELISA and PCR.

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Authors: A. Sharma, R. Tewari, S. K. Soni

Abstract:

Biofuels production has come forth as a future technology to combat the problem of depleting fossil fuels. Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient method and catching the eye of scientific community. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus main objective of the present study was to optimize composition of medium components for enhancing cellulase production by newly isolated strain of Bacillus tequilensis. Nineteen factors were taken into account using statistical Plackett-Burman Design. The significant variables influencing the cellulose production were further employed in statistical Response Surface Methodology using Central Composite Design for maximizing cellulase production. The optimum medium composition for cellulase production was: peptone (4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L), Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride (0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at 37oC. Analysis of variance (ANOVA) revealed high coefficient of determination (R2) of 0.99. Maximum cellulase productivity of 11.5 IU/ml was observed against the model predicted value of 13 IU/ml. This was found to be optimally active at 60oC and pH 5.5.

Keywords: Bacillus tequilensis, CMCase, Submerged Fermentation, Optimization, Plackett-Burman Design, Response Surface Methodology.

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Authors: Mário Silva, Filipa Gomes, Filipa Oliveira, Simone Morais, Cristina Delerue-Matos

Abstract:

Brown seaweeds are abundant in Portuguese coastline and represent an almost unexploited marine economic resource. One of the most common species, easily available for harvesting in the northwest coast, is Saccorhiza polyschides grows in the lowest shore and costal rocky reefs. It is almost exclusively used by local farmers as natural fertilizer, but contains a substantial amount of valuable compounds, particularly alginates, natural biopolymers of high interest for many industrial applications. Alginates are natural polysaccharides present in cell walls of brown seaweed, highly biocompatible, with particular properties that make them of high interest for the food, biotechnology, cosmetics and pharmaceutical industries. Conventional extraction processes are based on thermal treatment. They are lengthy and consume high amounts of energy and solvents. In recent years, microwave-assisted extraction (MAE) has shown enormous potential to overcome major drawbacks that outcome from conventional plant material extraction (thermal and/or solvent based) techniques, being also successfully applied to the extraction of agar, fucoidans and alginates. In the present study, acid pretreatment of brown seaweed Saccorhiza polyschides for subsequent microwave-assisted extraction (MAE) of alginate was optimized. Seaweeds were collected in Northwest Portuguese coastal waters of the Atlantic Ocean between May and August, 2014. Experimental design was used to assess the effect of temperature and acid pretreatment time in alginate extraction. Response surface methodology allowed the determination of the optimum MAE conditions: 40 mL of HCl 0.1 M per g of dried seaweed with constant stirring at 20ºC during 14h. Optimal acid pretreatment conditions have enhanced significantly MAE of alginates from Saccorhiza polyschides, thus contributing for the development of a viable, more environmental friendly alternative to conventional processes.

Keywords: Acid pretreatment, Alginate, Brown seaweed, Microwave-assisted extraction, Response surface methodology.

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Authors: S. S. Rana, C. Janveja, S. K. Soni

Abstract:

This study is concerned with the optimization of fermentation parameters for the hyper production of mannanase from Fusarium oxysporum SS-25 employing two step statistical strategy and kinetic characterization of crude enzyme preparation. The Plackett-Burman design used to screen out the important factors in the culture medium revealed 20% (w/w) wheat bran, 2% (w/w) each of potato peels, soyabean meal and malt extract, 1% tryptone, 0.14% NH4SO4, 0.2% KH2PO4, 0.0002% ZnSO4, 0.0005% FeSO4, 0.01% MnSO4, 0.012% SDS, 0.03% NH4Cl, 0.1% NaNO3 in brewer’s spent grain based medium with 50% moisture content, inoculated with 2.8×107 spores and incubated at 30oC for 6 days to be the main parameters influencing the enzyme production. Of these factors, four variables including soyabean meal, FeSO4, MnSO4 and NaNO3 were chosen to study the interactive effects and their optimum levels in central composite design of response surface methodology with the final mannanase yield of 193 IU/gds. The kinetic characterization revealed the crude enzyme to be active over broader temperature and pH range. This could result in 26.6% reduction in kappa number with 4.93% higher tear index and 1% increase in brightness when used to treat the wheat straw based kraft pulp. The hydrolytic potential of enzyme was also demonstrated on both locust bean gum and guar gum.

Keywords: Brewer’s Spent Grain, Fusarium oxysporum, Mannanase, Response Surface Methodology.

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Authors: Sanaa K. Bardaweel, Mohammad M. Hudaib, Khaled A. Tawaha, Rasha M. Bashatwah

Abstract:

Eucalyptus species are well reputed for their traditional use in Asia as well as in other parts of the world; therefore, the present study was designed to investigate the antimicrobial and antioxidant activities associated with essential oils from different Eucalyptus species. Essential oils from the leaves of six Eucalyptus species, including: Eucalyptus woodwardi, Eucalyptus stricklandii, Eucalyptus salubris, Eucalyptus sargentii, Eucalyptus torquata and Eucalyptus wandoo were separated by hydrodistillation and dried over anhydrous sodium sulphate. DPPH, ferric reducing antioxidant power, and hydroxyl radical scavenging activity assays were carried out to evaluate the antioxidant potential of the oils. The results indicate that examined oils exhibit substantial antioxidant activities relative to ascorbic acid. Previously, these oils were evaluated for their antimicrobial activities, against wide range of bacterial and fungal strains, and they were shown to possess significant antimicrobial activities. In this study, further investigation into the growth kinetics of oil-treated microbial cultures was conducted. The results clearly demonstrate that the microbial growth was markedly inhibited when treated with sub-MIC concentrations of the oils. Taken together, the results obtained indicate a high potential of the examined essential oils as bioactive oils, for nutraceutical and medical applications, possessing significant antioxidant and anti microbial activities.

Keywords: Antimicrobial, antioxidants, essential (volatile) oil, Eucalyptus.

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Authors: Kobra Nalbandi, Bahram Baghban Kohnehrouz, Khalil Alami Saeed

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The low level of foreign genes expression in transgenic plants is a key factor that limits plant genetic engineering. Because of the critical regulatory activity of the promoters on gene transcription, they are studied extensively to improve the efficiency of the plant transgenic system. The strong constitutive promoters, such as CaMV 35S promoter and Ubiqutin 1 maize are usually used in plant biotechnology research. However the expression level of the foreign genes in all tissues is often undesirable. But using a strong seed-specific promoter to limit gene expression in the seed solves such problems. The purpose of this study is to isolate one of the seed specific promoters of Hordeum vulgare. So one of the common varieties of Hordeum vulgare in Iran was selected and their genomes extracted then the D-Hordein promoter amplified using the specific designed primers. Then the amplified fragment of the insert cloned in an appropriate vector and then transformed to E. coli. At last for the final admission of accuracy the cloned fragments sent for sequencing. Sequencing analysis showed that the cloned fragment DHPcontained motifs; like TATA box, CAAT-box, CCGTCC-box, AMYBOX1 and E-box etc., which constituted the seed-specific promoter activity. The results were compared with sequences existing in data banks. D-Hordein promoters of Alger has 99% similarity at 100 % coverage. The results also showed that D-Hordein promoter of barley and HMW promoter of wheat are too similar.

Keywords: Barley, Seed specific promoter, Hordein.

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Authors: Karuppaiya Maruthai, Viruthagiri Thangavelu, Manikandan Kanagasabai

Abstract:

In the present study, effect of critical medium components (a total of fifteen components) on ethanol production from waste cashew apple juice (CAJ) using yeast Saccharomyces diasticus was studied. A statistical response surface methodology (RSM) based Plackett-Burman Design (PBD) was used for the design of experiments. The design contains a total of 32 experimental trails. The effect of medium components on ethanol was studied at two different levels such as low concentration level (-) and high concentration levels (+). The dependent variables selected in this study were ethanol concentration (g/L) and cellmass concentration (g/L). Data obtained from RSM on ethanol production were subjected to analysis of variance (ANOVA). In general, initial substrate concentration significantly influenced the microbial growth and product formation. Of the medium components evaluated, CAJ concentration, yeast extract, (NH4)2SO4, and malt extract showed significant effect on ethanol fermentation. A second-order polynomial model was used to predict the experimental data and the model fitted the data with a high correlation coefficient (R2 > 0.98). Maximum ethanol (15.3 g/L) and biomass (6.4 g/L) concentrations were obtained at the optimum medium composition and at optimum condition (temperature-30°C; initial pH-6.8) after 72 h fermentation using S.diasticus.

Keywords: cashew apple juice, ethanol, fermentation, yeast, response surface methodology

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Authors: N. P. Dhull, K. Gupta, R. Soni, D. K. Rahi, S. K. Soni

Abstract:

The aim of the present work was to statistically design an autotrophic medium for maximum biomass production by Chlorella pyrenoidosa using response surface methodology. After evaluating one factor at a time approach, K2HPO4, KNO3, MgSO4.7H2O and NaHCO3 were preferred over the other components of the fog’s medium as most critical autotrophic medium components. The study showed that the maximum biomass yield was achieved while the concentrations of MgSO4.7H2O, K2HPO4, KNO3 and NaHCO3 were 0.409 g/L, 0.24 g/L, 1.033 g/L, and 3.265 g/L, respectively. The study reported that the biomass productivity of C. pyrenoidosa improved from 0.14 g/L in defined fog’s medium to 1.40 g/L in modified fog’s medium resulting 10 fold increase. The biochemical composition biosynthesis of C. pyrenoidosa was altered using nitrogen limiting stress bringing about 5.23 fold increase in lipid content than control (cell without stress), as analyzed by FTIR integration method.

Keywords: Autotrophic condition, Chlorella pyrenoidosa, FTIR, Response Surface Methodology, Optimization.

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Authors: T. S. Moe, T. A. Khaing

Abstract:

Chitosan is a derivative of chitin, a compound usually isolated from the shells of some crustaceans such as crab, lobster and shrimp. It has biocompatible, biodegradable, and antimicrobial properties. To use these properties of chitosan in biomedical fields, chitosan films (1%, 2%, 3% and 4%) were prepared by using l% lactic acid as solvent. The effects of chitosan films on tensile strength, elongation at break, degree of swelling, thickness, morphology, allergic and irritation reactions and antibacterial property were evaluated. Staphylococcus aureus and Escherichia coli were used as tested microorganisms. In vivo wound healing activities of chitosan films were investigated using mice model. As results, Chitosan films have similar appearance and good swelling properties and 4% chitosan film showed the better swelling activity and the greatest elongation ratio than the other chitosan films. They also showed their good activity of wound healing in mice model. Moreover, the results showed that the films did not produce any unwilling symptoms (allergy or irritation). In conclusion, it is evident that the chitosan film has the potentiality to use as wound healing biofilms in the biomedical fields.

Keywords: Chitosan, wound healing, antibacterial activity.

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Authors: M. M. Seif, F. A. Khalil, A. A. K. Abou Arab, A. S. Abdel- Aziz, M. A. Abou Donia, Sh. R. Mohamed

Abstract:

Malathion (ML) is a well known pesticide commonly used in many agricultural and non-agricultural processes. Its toxicity has been attributed primarily to the accumulation of acetylcholine (Ach) at nerve junctions, due to the inhibition of acetylcholinesterase (AChE). The aim of the current research was to study the protective effect of the melissa plant extract against reproductive impairment induced by malathion in 32 male albino rats, and the biological experiment was divided into four groups (8 in each) that given malathion (27 mg/kg; 1/50 of the LD50 for an oral dose) and/or Melissa officinalis (MO) extract (200mg/kg/day) by gavages technique. The sperm counts, sperm motility, sperm morphology, FSH, LH, and testosterone levels had been determined in testes homogenate at the end of the experiment. It is worthy to report that, rats treated with melissa extract did not show a significant difference when compared with the control group, while rats given malathion alone had significantly lower sperm count, sperm motility, and significantly higher abnormal sperm numbers, than the untreated control rats as well as having significantly lower serum FSH, LH, and testosterone levels compared with the control group. Administrations of melissa extract restore all mentioned histological parameters towards the control group and the melissa extract had a strong positive protective effect against malathion toxicity. Results the of biological parameters were confirmed by the histological examination of rat testes and indicated that, both control and melissa groups showing normal seminiferous tubules, while malathion group testicular tissues had necrosis, edema in the seminiferous tubules and degeneration of spermatogonial cells lining the seminiferous tubules with incomplete spermatogenesis. The use of melissa against malathion improved the histological picture and showing normal seminiferous tubules with complete spermatogenesis and almost there was no histopathological changes could be noted.

Keywords: Malathion, Melissa officinalis L., Reproductive toxicity, Rats.

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Authors: Abou-Baker Salim, A. A. K. Abou-Arab, Sherif R. Mohamed, T. A. Eldesouky

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Pomegranate (Punica granatum L.) is an ancient fruit of great medical interest and rich source of antioxidants. Pesticides as dimethoate play a crucial role in the occurrence many diseases in plants, animal and human. Therefore the ability of Pomegranate (Punica granatum L.) to alleviate hepatotoxicity induced by organophosphate pesticide dimethoate was investigated. Albino male rats were divided randomly into 4 groups and kept at 7 animals per group in an environmentally controlled condition for 6 weeks. The first group was served as a control group (basal diet), the second group fed on basal diet supplemented with 5% freeze dried pomegranate seeds, the third group fed on 20 ppm dimethoate contaminated diet and the last group fed on dimethoate contaminated diet supplemented with 5% freeze dried pomegranate seeds. The results revealed that administration of dimethoate caused high significant increased in liver functions: alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) activities as well as lipid peroxide (malonaldhyde, MDA); on the other hand high significant decreased on glutathione (GSH), glutathione peroxidase (GPx), albumin and total protein were observed. However addition of 5% freeze dried pomegranate seeds significantly improved all previously mentioned parameters. These results indicate the dimethoate induced hepatotoxicity and highlight the protective effect of pomegranate seeds as a potential protective agent against dimethoate induced hepatotoxicity. This may be attributed to the powerful antioxidants (polyphenols, total phenols, and total flavonoids) which present in high levels in pomegranate as well as improving the immunity by activation of antioxidant enzymes GSH and GPx.

Keywords: Pomegranate, organophosphate pesticides, dimethoate, liver toxicity, free radicals, antioxidants.

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Authors: R. Nor Azah, A. R. Roshanida, N. Norzita

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The biomass-based fuels have become great concern in order to replace the petroleum-based fuels. Biofuels are a wide range of fuels referred to liquid, gas and solid fuels produced from biomass. Recently, higher chain alcohols such as 3-methyl-1-butanol and isobutanol have become a better candidate compared to bioethanol in order to replace gasoline as transportation fuel. Therefore, in this study, 3-methyl-1-butanol was produced through a fermentation process by yeast. Several types of yeast involved in this research including Saccharomyces cerevisiae, Kluyveromyces lactis GG799 and Pichia pastoris (KM71H, GS115 and X33). The result obtained showed that K. lactis GG799 gave the highest concentration of 3-methyl-1-butanol at 274 mg/l followed by S. cerevisiae, P. pastoris GS115, P. pastoris KM71H and P. pastoris X33 at 265 mg/l, 190 mg/l, 182 mg/l and 174 mg/l respectively. Based on the result, it proved that yeast have a potential in producing 3-methyl-1-butanol naturally.

Keywords: Biofuel, fermentation, 3-methyl-1-butanol, yeast.

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Authors: Seyed Ali Jafari, Mohammad Ghomi Avili, Emad Benhelal

Abstract:

In this study, a mathematical model was proposed and the accuracy of this model was assessed to predict the growth of Pseudomonas aeruginosa and rhamnolipid production under nitrogen limiting (sodium nitrate) fed-batch fermentation. All of the parameters used in this model were achieved individually without using any data from the literature. The overall growth kinetic of the strain was evaluated using a dual-parallel substrate Monod equation which was described by several batch experimental data. Fed-batch data under different glycerol (as the sole carbon source, C/N=10) concentrations and feed flow rates were used to describe the proposed fed-batch model and other parameters. In order to verify the accuracy of the proposed model several verification experiments were performed in a vast range of initial glycerol concentrations. While the results showed an acceptable prediction for rhamnolipid production (less than 10% error), in case of biomass prediction the errors were less than 23%. It was also found that the rhamnolipid production by P. aeruginosa was more sensitive at low glycerol concentrations. Based on the findings of this work, it was concluded that the proposed model could effectively be employed for rhamnolipid production by this strain under fed-batch fermentation on up to 80 g l- 1 glycerol.

Keywords: Fed-batch culture, glycerol, kinetic parameters, modelling, Pseudomonas aeruginosa, rhamnolipid.

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Authors: Emma K. Sales, Nilda G. Butardo

Abstract:

The project was undertaken to determine the effects of modified tissue culture protocols e.g. age of culture and hormone levels (2,4-D) in generating somaclonal variation. Moreover, the utility of molecular markers (SSR and MSAP) in sorting off types/somaclones were investigated.

Results show that somaclonal variation is in effect due to prolonged subculture and high 2,4-D concentration. The resultant variation was observed to be due to high level of methylation events specifically cytosine methylation either at the internal or external cytosine and was identified by methylation sensitive amplification polymorphism (MSAP).Simple sequence repeats (SSR) on the other hand, was able to associate a marker to a trait of interest.

These therefore, show that molecular markers can be an important tool in sorting out variation/mutants at an early stage.

Keywords: Methylation, MSAP, somaclones, SSR, subculture, 2, 4-D.

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Authors: Rosa Buxeda, Lorenzo Saliceti-Piazza, Rodolfo J. Romañach, Luis Ríos, Sandra L. Maldonado-Ramírez

Abstract:

Biopharmaceuticals manufacturing is one of the major economic activities worldwide. Ninety-three percent of the workforce in a biomanufacturing environment concentrates in production-related areas. As a result, strategic collaborations between industry and academia are crucial to ensure the availability of knowledgeable workforce needed in an economic region to become competitive in biomanufacturing. In the past decade, our institution has been a key strategic partner with multinational biotechnology companies in supplying science and engineering graduates in the field of industrial biotechnology. Initiatives addressing all levels of the educational pipeline, from K-12 to college to continued education for company employees have been established along a ten-year span. The Amgen BioTalents Program was designed to provide undergraduate science and engineering students with training in biomanufacturing. The areas targeted by this educational program enhance their academic development, since these topics are not part of their traditional science and engineering curricula. The educational curriculum involved the process of producing a biomolecule from the genetic engineering of cells to the production of an especially targeted polypeptide, protein expression and purification, to quality control, and validation. This paper will report and describe the implementation details and outcomes of the first sessions of the program.

Keywords: Biomanufacturing curriculum, interdisciplinary learning, workforce development, industry-academia partnering.

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Authors: V. Chahal, A. Nagpal, Y. B. Pakade, J. K. Katnoria

Abstract:

Soil is a complex physical and biological system that provides support, water, nutrients and oxygen to the plants. Apart from these, it acts as a connecting link between inorganic, organic and living components of the ecosystem. In recent years, presence of xenobiotics, alterations in the natural soil environment, application of pesticides/inorganic fertilizers, percolation of contaminated surface water as well as leachates from landfills to subsurface strata and direct discharge of industrial wastes to the land have resulted in soil pollution which in turn has posed severe threats to human health especially in terms of causing carcinogenicity by direct DNA damage. The present review is an attempt to summarize literature on sources of soil pollution, characterization of pollutants and their consequences in different living systems.

Keywords: Soil Pollution, Contaminants, Heavy metals, Pesticides, Bioassays.

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Authors: Ratanaporn Leesing, Thidarat Papone, Mutiyaporn Puangbut

Abstract:

Mixotrophic cultivation of the isolated freshwater microalgae Chlorella sp. KKU-S2 in batch shake flask for biomass and lipid productions, different concentration of glucose as carbon substrate, different nitrogen source and concentrations were investigated. Using 1.0g/L of NaNO₃ as nitrogen source, the maximum biomass yield of 10.04g/L with biomass productivity of 1.673g/L d was obtained using 40g/L glucose, while a biomass of 7.09, 8.55 and 9.45g/L with biomass productivity of 1.182, 1.425 and 1.575g/L d were found at 20, 30 and 50g/L glucose, respectively. The maximum lipid yield of 3.99g/L with lipid productivity of 0.665g/L d was obtained when 40g/L glucose was used. Lipid yield of 1.50, 3.34 and 3.66g/L with lipid productivity of 0.250, 0.557 and 0.610g/L d were found when using the initial concentration of glucose at 20, 30 and 50g/L, respectively. Process product yield (Y_P/S) of 0.078, 0.119, 0.158 and 0.094 were observed when glucose concentration was 20, 30, 40 and 50 g/L, respectively. The results obtained from the study shows that mixotrophic culture of Chlorella sp. KKU-S2 is a desirable cultivation process for microbial lipid and biomass production. 

Keywords: Mixotrophic cultivation, microalgal lipid, Chlorella sp. KKU-S2.

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Authors: Surasak Siripornadulsil, Nutt Poomai, Wilailak Siripornadulsil

Abstract:

Due to a high ethanol demand, the approach for  effective ethanol production is important and has been developed  rapidly worldwide. Several agricultural wastes are highly  abundant in celluloses and the effective cellulase enzymes do exist  widely among microorganisms. Accordingly, the cellulose  degradation using microbial cellulase to produce a low-cost substrate  for ethanol production has attracted more attention. In this  study, the cellulase producing bacterial strain has been isolated  from rich straw and identified by 16S rDNA sequence analysis as Acinetobacter sp. KKU44. This strain is able to grow and exhibit the cellulase activity. The optimal temperature for its growth and  cellulase production is 37°C. The optimal temperature of bacterial  cellulase activity is 60°C. The cellulase enzyme from  Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36h. showed highest cellulase activity at 120U/mL when  grown in LB medium containing 2% (w/v). The capability of  Acinetobacter sp. KKU44 to grow in cellulosic agricultural wastes as a sole carbon source and exhibiting the high cellulase activity at high temperature suggested that this strain could be potentially developed further as a cellulose degrading strain for a production of low-cost substrate used in ethanol production. 

 

Keywords: Acinetobacter sp. KKU44, bagasse, cellulase enzyme, rice husk.

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Authors: Soni Yadav, Sitansu Kumar Verma, Jitendra Singh, Ajay Kumar

Abstract:

This article comprises detail information about L-asparaginase, encompassing topic such as various sources of L-asparaginase, mechanism and properties of L-asparaginase. Also describe the production, cultivation and purification of L-asparaginase along with information about the application of L-asparaginase. L-asparaginase catalyzes the conversion reaction to convert asparagine to aspartic acid and ammonia. Asparagine is a nutritional requirement for both normal and tumor cell. Present scenario has found that L-asparaginase has been found to be a best anti tumor or antileukemic agent. In the recent years this enzyme gained application in the field of clinical research pharmacologic and food industry. It has been characterized based on the enzyme assay principle hydrolyzing L-asparagine into L-aspartic acid and ammonia. It has been observed that eukaryotic microorganisms such as yeast and filamentous fungi have a potential for L-asparaginase production. L-asparaginase has been and is still one of the most lengthily studied therapeutic enzymes by scientist and researchers worldwide.

Keywords: L-asparaginase, antitumor, solid state fermentation, chemotherapeutic.

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Authors: E. F. Aransiola, M.O. Daramola, E. O. Iwalewa, A. M. Seluwa, O. O. Olufowobi

Abstract:

Diabetes is a chronic metabolic disorder that affects the quality of life in terms of physical health, social and psychological well-being. In spite of the enormous progress in the treatment of diabetes using existing commercial drugs, such as, insulin and oral hypoglycemic agents, the quest and search for new drugs is imperative due to several limitations of the commercial drugs. In addition, the existing diabetic drugs are expensive and unaffordable by the rural populace in the developing countries. The present study demonstrates the anti-diabetic property of aqueous extract of Bryophyllum pinnatum (BP) leaves using diabetic rats (albino rats) as models. At the same time, the anti-diabetic effect of the aqueous extract was compared to that of a sample containing a mixture of the extract and a commercial diabetic medicine, glibenclamide. A specified dosage of aqueous extract of Bryophyllum pinnatum (BP) leaves was administered on the experimental diabetic rats, and their BGL was measured and recorded. The results showed a significant drop in the BGL of the diabetic rats to a value close to normal blood glucose level within 120 minutes when only aqueous extract from BP leaves was used. When a sample containing a mixture of the aqueous extract and glibenclamide was administered, a further drop in BGL was observed. Therefore, the results reveal that aqueous extract of Bryophyllum pinnatum leaves have significant anti-diabetic properties, and that the performance of the existing drugs (glibenclamide) could be enhanced with the use of the aqueous extract.

Keywords: Anti-diabetics, Bryophyllum pinnatum, Blood glucose level, albino rats.

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