In order to clean up soils contaminated with hydrocarbons, the bioremediation activity of Pseudomonas putida was studied. Pseudomonas putida is a bacterium that can withstand the harshest environmental conditions. It is able to metabolize a wide range of petroleum hydrocarbons which is used as a source of carbon and energy. Given the potential of this microorganism, an experiment wasconducted on this strain.
For the isolation of this microorganism, a sample ofsoil from the Vakinankaratra region in the urban commune of Antsirabe II, Madagascar was microbiologically analysed. The bacterial identification was based on a study of the morphological, physicochemical and sequential analysis of the 16S rDNA gene.
Low density polythene (LDPE) is the most widely used packaging material primarily because of its excellent mechanical properties, barrier properties against water, light weight, low cost and high energy effectiveness.
LDPE to biological attack was believed to be contributed by the hydrophobic carbon backbone and high molecular weight of the polymer. Thus, over the years, the rapid biodegradation of plastic has been a subject of interest in the waste management problem.
Each year, an estimated 500 billion to 1 trillion plastic bags are consumed worldwide. After their use, these packaging materials are dumped in landfills leading to pollution since they are non-biodegradable under natural environmental conditions
Soil organic matter has long been recognized as one of the most important components in maintaining soil fertility, soil quality, and agricultural sustainability. The soil zone strongly influenced by plant roots, the rhizosphere, plays an important role in regulating soil organic matter decomposition and nutrient cycling. Processes that are largely controlled or directly influenced by roots are often referred to as rhizosphere processes. These processes may include exudation of soluble compounds, water uptake, nutrient mobilization by roots and microorganisms, rhizosphere-mediated soil organic matter decomposition, and the subsequent release of CO2 through respiration. Rhizosphere processes are major gateways for nutrients and water. At the global scale, rhizosphere processes utilize approximately 50% of the energy fixed by photosynthesis in terrestrial ecosystems, contribute roughly 50% of the total CO2 emitted from terrestrial ecosystems, and mediate virtually all aspects of nutrient cycling. Therefore, plant roots and their rhizosphere interactions are at the center of many ecosystem processes. However, the linkage between rhizosphere processes and soil organic matter decomposition is not well understood. Because of the lack of appropriate methods, rates of soil organic matter decomposition are commonly assessed by incubating soil samples in the absence of vegetation and live roots with an implicit assumption that rhizosphere processes have little impact on the results. Our recent studies have overwhelmingly proved that this implicit assumption is often invalid, because the rate of soil organic matter decomposition can be accelerated by as much as 380% or inhibited by as much as 50% by the presence of live roots. The rhizosphere effect on soil organic matter decomposition is often large in magnitude and significant in mediating plant-soil interactions.
In this slide different fungi are Mentioned and their role as bio-control agents is also elaborated which is reviewed from different research articles cited in reference portion.
An entomopathogenic fungus can act as a parasite of insects and kills or seriously disables them.Targets are distributed among 10 insect orders:
Hemiptera (59.6%), Coleoptera (40.9%), Lepidoptera (17.5%), Thysanoptera (14.6%), Orthoptera (9.4%), Diptera (7.0%), Hymenoptera (2.9%), Isoptera (2.3%), Siphonoptera (1.2%), and Blattodea(0.6%).
Low density polythene (LDPE) is the most widely used packaging material primarily because of its excellent mechanical properties, barrier properties against water, light weight, low cost and high energy effectiveness.
LDPE to biological attack was believed to be contributed by the hydrophobic carbon backbone and high molecular weight of the polymer. Thus, over the years, the rapid biodegradation of plastic has been a subject of interest in the waste management problem.
Each year, an estimated 500 billion to 1 trillion plastic bags are consumed worldwide. After their use, these packaging materials are dumped in landfills leading to pollution since they are non-biodegradable under natural environmental conditions
Soil organic matter has long been recognized as one of the most important components in maintaining soil fertility, soil quality, and agricultural sustainability. The soil zone strongly influenced by plant roots, the rhizosphere, plays an important role in regulating soil organic matter decomposition and nutrient cycling. Processes that are largely controlled or directly influenced by roots are often referred to as rhizosphere processes. These processes may include exudation of soluble compounds, water uptake, nutrient mobilization by roots and microorganisms, rhizosphere-mediated soil organic matter decomposition, and the subsequent release of CO2 through respiration. Rhizosphere processes are major gateways for nutrients and water. At the global scale, rhizosphere processes utilize approximately 50% of the energy fixed by photosynthesis in terrestrial ecosystems, contribute roughly 50% of the total CO2 emitted from terrestrial ecosystems, and mediate virtually all aspects of nutrient cycling. Therefore, plant roots and their rhizosphere interactions are at the center of many ecosystem processes. However, the linkage between rhizosphere processes and soil organic matter decomposition is not well understood. Because of the lack of appropriate methods, rates of soil organic matter decomposition are commonly assessed by incubating soil samples in the absence of vegetation and live roots with an implicit assumption that rhizosphere processes have little impact on the results. Our recent studies have overwhelmingly proved that this implicit assumption is often invalid, because the rate of soil organic matter decomposition can be accelerated by as much as 380% or inhibited by as much as 50% by the presence of live roots. The rhizosphere effect on soil organic matter decomposition is often large in magnitude and significant in mediating plant-soil interactions.
In this slide different fungi are Mentioned and their role as bio-control agents is also elaborated which is reviewed from different research articles cited in reference portion.
An entomopathogenic fungus can act as a parasite of insects and kills or seriously disables them.Targets are distributed among 10 insect orders:
Hemiptera (59.6%), Coleoptera (40.9%), Lepidoptera (17.5%), Thysanoptera (14.6%), Orthoptera (9.4%), Diptera (7.0%), Hymenoptera (2.9%), Isoptera (2.3%), Siphonoptera (1.2%), and Blattodea(0.6%).
he rhizosphere is the narrow region of soil or substrate that is directly influenced by root secretions and associated soil microorganisms known as the root microbiome.
The phyllosphere is a term used in microbiology to refer to the total above-ground portions of plants as habitat for microorganisms.
Lignocelluloses, the major component of biomass, makes up about half of the matter produced by photosynthesis. It consists of three types of polymers – cellulose, hemicellulose, and lignin – that are strongly intermeshed and chemically bonded by non-covalent forces and by covalent cross-linkages. A great variety of fungi and bacteria can fragment these macromolecules by using a battery of hydrolytic or oxidative enzymes. In native substrates, binding of the polymers hinders their biodegradation. Molecular genetics of cellulose-, hemicellulose- and lignin-degrading systems advanced considerably during the 1990s. Most of the enzymes have been cloned, sequenced, and expressed both in homologous and in heterologous hosts. Much is known about the structure, genomic organization, and regulation of the genes encoding these proteins.
Molecular Characterization of Isolated Methyl Parathion Degrading Bacteria an...IJSRD
The burning problem in present era is pesticide residue in fruits and foods. present study focus on degradation of pesticide in contest to that total 45 methyl parathion(MP) degrading bacteria were isolated from sludge of MP producing industrial waste. According to screening 21 highly efficient pesticide degraders were selected at 500ppm concentration of standard methyl parathion as a sole carbon source. All isolates were characterized by RAPD and based on the RAPD result all 21 bacterial isolates were grouped in to 2 main clusters with 58% similarity. Characterization of MP degrading gene was carried out by using specific primer. Out of 21 isolates, 14 isolates were containing mpd gene and 4 isolates were containing opd gene while remaining 3 isolates which did not show amplification with opd/ mpd primer so it may contain other unreported gene responsible for methyl parathion degradation. Isolates were identified based on 16s rRNA sequence and one highly efficient bacterium selected for genome characterization.
Abstract
Objective(s):
Biosynthesis of gold nanoparticles (NGPs) is environmentally safer than chemical and physical procedures. This method requires no use of toxic solvents and synthesis of dangerous products and is environmentally safe. In this study, we report the biosynthesis of NGPs using Streptomyces djakartensis
isolate B-5.
Materials and Methods:
NGPs were biosynthesized by reducing aqueous gold chloride solution via a Streptomyces isolate without the need for any additive for protecting nanoparticles from aggregation. We characterized the responsible Streptomycete; its genome DNA was isolated, purified and 16S rRNA was amplified by PCR. The amplified isolate was sequenced; using the BLAST search tool from NCBI, the microorganism was identified to species level.
Results:
Treating chloroauric acid solutions with this bacterium resulted in reduction of gold ions and formation of stable NGPs. TEM and SEM electro micrographs of NGPs indicated size range from 2- 25 nm with average of 9.09 nm produced intracellular by the bacterium. SEM electro micrographs revealed morphology of spores and mycelia. The amplified PCR fragment of 16S rRNA gene was cloned and sequenced from both sides; it consisted of 741 nucleotides. According to NCBI GenBank, the bacterium had 97.1% homology with Streptomyces djakartensis strain RT-49. The GenBank accession number for partial 16S rRNA gene was recorded as JX162550.
Conclusion:
Optimized application of such findings may create applications of Streptomycetes for use as bio-factories in eco-friendly production of NGPs to serve in demanding industries and related biomedical areas. Research in this area should also focus on the unlocking the full mechanism of NGPs biosynthesis by Streptomycetes.
he rhizosphere is the narrow region of soil or substrate that is directly influenced by root secretions and associated soil microorganisms known as the root microbiome.
The phyllosphere is a term used in microbiology to refer to the total above-ground portions of plants as habitat for microorganisms.
Lignocelluloses, the major component of biomass, makes up about half of the matter produced by photosynthesis. It consists of three types of polymers – cellulose, hemicellulose, and lignin – that are strongly intermeshed and chemically bonded by non-covalent forces and by covalent cross-linkages. A great variety of fungi and bacteria can fragment these macromolecules by using a battery of hydrolytic or oxidative enzymes. In native substrates, binding of the polymers hinders their biodegradation. Molecular genetics of cellulose-, hemicellulose- and lignin-degrading systems advanced considerably during the 1990s. Most of the enzymes have been cloned, sequenced, and expressed both in homologous and in heterologous hosts. Much is known about the structure, genomic organization, and regulation of the genes encoding these proteins.
Molecular Characterization of Isolated Methyl Parathion Degrading Bacteria an...IJSRD
The burning problem in present era is pesticide residue in fruits and foods. present study focus on degradation of pesticide in contest to that total 45 methyl parathion(MP) degrading bacteria were isolated from sludge of MP producing industrial waste. According to screening 21 highly efficient pesticide degraders were selected at 500ppm concentration of standard methyl parathion as a sole carbon source. All isolates were characterized by RAPD and based on the RAPD result all 21 bacterial isolates were grouped in to 2 main clusters with 58% similarity. Characterization of MP degrading gene was carried out by using specific primer. Out of 21 isolates, 14 isolates were containing mpd gene and 4 isolates were containing opd gene while remaining 3 isolates which did not show amplification with opd/ mpd primer so it may contain other unreported gene responsible for methyl parathion degradation. Isolates were identified based on 16s rRNA sequence and one highly efficient bacterium selected for genome characterization.
Abstract
Objective(s):
Biosynthesis of gold nanoparticles (NGPs) is environmentally safer than chemical and physical procedures. This method requires no use of toxic solvents and synthesis of dangerous products and is environmentally safe. In this study, we report the biosynthesis of NGPs using Streptomyces djakartensis
isolate B-5.
Materials and Methods:
NGPs were biosynthesized by reducing aqueous gold chloride solution via a Streptomyces isolate without the need for any additive for protecting nanoparticles from aggregation. We characterized the responsible Streptomycete; its genome DNA was isolated, purified and 16S rRNA was amplified by PCR. The amplified isolate was sequenced; using the BLAST search tool from NCBI, the microorganism was identified to species level.
Results:
Treating chloroauric acid solutions with this bacterium resulted in reduction of gold ions and formation of stable NGPs. TEM and SEM electro micrographs of NGPs indicated size range from 2- 25 nm with average of 9.09 nm produced intracellular by the bacterium. SEM electro micrographs revealed morphology of spores and mycelia. The amplified PCR fragment of 16S rRNA gene was cloned and sequenced from both sides; it consisted of 741 nucleotides. According to NCBI GenBank, the bacterium had 97.1% homology with Streptomyces djakartensis strain RT-49. The GenBank accession number for partial 16S rRNA gene was recorded as JX162550.
Conclusion:
Optimized application of such findings may create applications of Streptomycetes for use as bio-factories in eco-friendly production of NGPs to serve in demanding industries and related biomedical areas. Research in this area should also focus on the unlocking the full mechanism of NGPs biosynthesis by Streptomycetes.
To study of the genetic variations among the Azospirillum lipoferu isolates u...ijsrd.com
Among free-living microorganisms, which can be practically used in agriculture, bacteria from the Azospirillum genus as well as other endophytes are nowadays thought of as the most active component of associative dinitrogen fixation. The investigation was carried out to study the characterization of Azospirillum lipoferu found in the soils of the ten agro-climatic zones which Karnataka, is classified. By using RAPD markers, 75 bands were scored out of which 78.6 % were found to be polymorphic. Statistical analysis of RAPD data enabled the classification of 10 Azospirillum isolates into two major groups. . In this, the cluster analysis based on 75 RAPD bands revealed that the ten A. lipoferu isolates examined clustered at a linkage distance of about 40 units on the dendrogram. There was no correlation between RAPD and geographical origin of isolates.
The Role of Cell Wall-Degrading Enzymes in the Development of Anthracnose Dis...Agriculture Journal IJOEAR
— The ability of Colletotrichumtruncatum CP2 in producing pectinolytic and cellulolytic enzymes was evaluated by shake flask fermentations. The results of enzymatic activity experiment indicated that PG was the first cell wall-degrading enzymes detected and the activities obtained were higher (0.24±0.10 U/mL) than other enzymes, which appeared later and in lower amount. After the cell wall was degraded by the action of PG, further degradation of the cell wall was affected by pectin methylesterases, pectin lyase, pectate lyase and cellulases. The disparity in enzymatic activity at different intervals may suggest their specific role for pathogenesis at proper timings.
Microbial DNA extracted from two soil samples collected from Beni-Suef and Kafr El-Sheikh were
subjected to PCR amplification with primers specific for 16S rDNA gene and cloned in linear pCR 2.1
plasmid vector. Recombinants were transformed into Escherichia coli competent cells. Sixty clone
inserts (30 from each soil sample) were sequenced and subjected to phylogenetic analyses. Forty
sequences of the sixty clones were affiliated with previously recognized bacterial groups. Thirty six of
these had closest relatives among cultured taxa and clustered primarily with three divisions containing
microrganisms commonly associated with soil: Proteobacteria, Gram-positive organisms, and
Cytophaga-Flexibacter-Bacteroides group. The results also showed the presence of one clone related to
Nirospira retrieved from Beni-Suef soil, one clone from Archaea kingdom retrieved from Kafr El-Sheikh
soil, and three clones affiliated to the newly described Holophaga-Acidobacterium phylum in both Beni-
Suef and Kafr El-Sheikh soils. Seven sequences grouped with known divisions but had closest relatives
among soil taxa known only from rDNA sequences analysis. Twelve clone sequences were distantly
related to known sequences. Many of these sequences may represent new bacterial divisions.
Isolation and Growth Kinetic Studies Of Bacillus Methylotrophicus P10 & P11 ...theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
The papers for publication in The International Journal of Engineering& Science are selected through rigorous peer reviews to ensure originality, timeliness, relevance, and readability.
Theoretical work submitted to the Journal should be original in its motivation or modeling structure. Empirical analysis should be based on a theoretical framework and should be capable of replication. It is expected that all materials required for replication (including computer programs and data sets) should be available upon request to the authors.
The International Journal of Engineering & Science would take much care in making your article published without much delay with your kind cooperation.
IJPCBS 2012, 2(1), 110-116 Kavya et al. ISSN: 2249-9504
110
INTERNATIONAL JOURNAL OF PHARMACEUTICAL, CHEMICAL AND BIOLOGICAL SCIENCES
Available online at www.ijpcbs.com
ISOLATION AND SCREENING OF STREPTOMYCES SP. FROM
CORINGA MANGROVE SOILS FOR ENZYME PRODUCTION AND
ANTIMICROBIAL ACTIVITY
M. Kavya Deepthi1*, M. Solomon Sudhakar1 and M. Nagalakshmi Devamma2 1Department of Biotechnology, Rajalakshmi Engineering College, Thandalam, 2Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pr Taadmesihln, aInddui,a I.n dia.
Microbial Production Of Alkaline Proteases And Evaluation Of Its Performances...Shafkat Shamim Rahman
A high alkaline protease producing bacterial strain was isolated and identified a local soil sample. The organism was gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus subtilis. It was also identified as B. subtilis with 99.9% identity by API 50 CHB. The enzyme hydrolyses a number of proteins including azocasein which suggests that it is an extracellular alkaline protease. The experimentally determined isoelectric point was 5.1 and the optimal enzyme activity was at 60°C and at pH 8.5. The esterase preferentially hydrolyzed short-chain fatty acids. Native enzyme preparations typically showed a Michaelis constant (Km) and Vmax of 0.40mM and 12,200 U mg)-1, respectively. This microbial enzyme was partially purified by ammonium sulfate fractionation, dialysis, DEAE cellulose chromatography and electrophoretic analysis. Enzyme purity was tested by SDS-PAGE. Quantitative estimation has shown that 40mL of culture supernatant could dehair 2×1 cm of leather completely in 9 hours. In future the tanneries will use a combination of chemical and enzymatic processes. In practical applications, protease is a useful enzyme for promoting the hydrolysis of proteins and showing significant industrial applications.
ABSTRACT- The development of human civilization throughout history has led to growing disruption of the natural
balance and the occurrence of different types of pollution. Environmental pollution with petroleum and petrochemical
products has been recognized as significant and serious problem. Diesel engine oil, which is one of the major products of
crude oil, constitutes a major source of pollution in our environment. Therefore diesel engine oil can enter into the
environment through wrecks of oil tankers carrying diesel oil, cleaning of diesel tanks by merchants, war ships carrying
diesel oil and motor mechanics. In present study the microorganisms utilising petrol and diesel oil as carbon source were
isolated and investigation of their characteristics towards the production of polyhydroxyalkanoates (PHA), which is now a
days well known as biodegradable polymer.
Key Words- Petrol and Diesel oil contamination, Bioremediation, Biodegradable bacterial polymer, Sudan
Black B staining, 16sr RNA sequencing
In order to study the WGS on an industrial scale at a low pressure, the modeling andsimulation of a WGS reactor operating at a pressure close to Patm and processing an industrial charge in the presence of a high temperature shift catalyst (Fe2O3/Cr2O3) were performed. The Profiles of the carbon monoxide conversion, temperature and pressure along the reactor were obtained. The effect of several operating parameters (inlet temperature, H2O/CO ratio) on the conversion of carbon monoxide along the reactor has been determined. The estimated catalytic mass to convert 60.5% of the carbon monoxide contained in the inlet is 170.76 t. The pressure drops in the reactor are not negligible and the maximum temperaturereached is without any harmful effect on the catalyst. The choice of an optimal inlet temperature and a high H2O/CO ratio improves the conversion of carbon monoxide.
As we are all aware,therecent discovery of the Higgs boson has revealed a highly massive particle, the value of which lies between 125and 126.5 GeV/c2.. According to the basic concepts of Quantum Mechanics, and in full compliance with the Uncertainty Principle and Yukawa intuitions, we were able to calculate the maximum limit of the Higgs boson‟s field of action. From the calculations show that the Higgs boson presents a range of action really very small, namely 9.8828∙10-16[cm], that is slightly smaller than 10-15[cm]. This value is justified by the considerable mass that the Higgs bosonacquires, in perfect agreement with the Uncertainty Principle.
The dependencies of total pressure, velocity, vorticity, turbulent length, turbulent dissipation, turbulent viscosity, turbulent energy and turbulent time of moving fluid from a straight pipe length of a circular cross section are presented in graphical and mathematical forms. Changing analysis of considered parameters was performed at mass flow rates of 0.45, 1.0 and 1.5 kg/s. A transition boundary of laminar flow of fluid to turbulent flow is at the distance of 2/5 of length from the inlet of the pipe (at accepted total length of the pipe of 1000 mm).
This paper considers the problem of magneto-hydrodynamics (MHD) Newtonian and non-Newtonian nano fluid flow passing on a magnetic sphere with mixed convection effect. Nano Fluid is a combination of liquid fluid as a base fluid with small solid nano particles. Water is chosen as Newtonian base fluid and oil is chosen as non-Newtonian base fluid. Then, Alumina and Copper are chosen as solid particle in nano fluid. We further construct governing equation by applying continuity equation, momentum equation, and energy equation to obtain dimensional governing equations. The dimensional governing equations that have been obtained are converted into non-dimensional governing equations by substituting non-dimensional variables. The non-dimensional governing equations are further transformed into similarity equations using stream function and solved numerically using Euler Implicit Finite Difference method. We further analyse the effect of magnetic parameter towards velocity and temperature in MHD nano fluid flow. The results show that the increases of magnetic parameter impacts to the decrease of velocity and temperature. Then, the velocity and temperature of Newtonian nano fluid are higher than the velocity and temperature of non-Newtonian nano fluid. Also, the velocity and temperature of copper-water are higher than the velocity and temperature of Alumina-water.
Building materials used for the walls of simple houses in lower-middle-class areas in Indonesia are currently dominated by brick. This study proposes that soil-paper blocks coated with calcium silicate board may be a suitable alternative, with high embodied energy and density. The research aims to obtain an optimal wall thickness to provide protection against cooling and embodied energy in low income houses, as well as against the temperature conditions in these buildings in highland and lowland areas. Determination of wall thickness is performed by simulation of a 9 m2 building model with thick variables. Cooling calculations involved the use of Archipak software. Temperature measurements were carried out using a data logger on a sample of soil-paper blocks. The results indicate that the optimal wall thickness for protection against cooling and embodied energy is 8 cm. Soil-paper block has a lower density than brick. The use of calcium silicate boards does not affect the internal temperature of a low income house, but they can be used as protection against rainwater and as a substitute for wall plastering.
Adaptive-optimal control involves re-identification of the machining process and the model obtained is used to calculate the optimal process parameters.
Optimal control characterizes the addiction of the technical and economic indicators to process parameters. Characteristic for performance technical indicators is that their dependence to parameter values of process has a limitative, what leads to one of the following conclusions, appropriately or inappropriately, and therefore can serve as restrictions in optimization problem.
Economic indicators have a continuous dependence of process parameters and therefore they are used as objective functions.
Knowledge management (KM) has become an effective way of managing organization‟s intellectual capital or, in other words, organization‟s full experience, skills and knowledge that is relevant for more effective performance in future. The paper proposes a knowledge management to achieve a competitive control of the machining systems. Then an application of Knowledge Management in engineering has been attempted to explain. The model can be used by the manager for the choosing of competitive orders.
Ceftriaxone is one of the third generations of cephalosporin antibiotics and commercially found as a sodium salt. The market demand for it is still high in recent years, including in Indonesia. However, there is no local production manufacture yet. A high yield of ceftriaxone sodium would be an advantage in industrial scale. Ceftriaxone was synthesized by reacting 7–amino–3–[(2,5–dihydro–6–hydroxy–2–methyl–5–oxo–1,2,4–triazin–3–yl) thiomethyl] cephalosporanic acid (7-ACT) with 2-Mercaptobenzothiazolyl (Z)-2-(2-Aminothiazole-4-yl)-2-Methoxyimino Acetate (MAEM) then with sodium salt in basic condition. The product was generated by solventing-out using acetone. The products were analyzed by HPLC quantitatively and the structure was confirmed using FTIR, MS and NMR. In this research, the variation in the mole ratio of reactants against the yield of product was evaluated. The result showed that the variations in mole ratio reactants affect the yield production. The higher ratio of MAEM would be the higher yield is obtained. The results show that the yield was 72,17% at mole ratio 1:2 which has 99,32% purity. This result could be a consideration in industrial production scale in ceftriaxone sodium preparation.
The challenges of river water quality management are so enormous, due to the unpredictive modes of contamination. Monitoring different sources of pollutant load contribution to the river basin is also quite tasking, resulting to laborious and expensive process which sometimes lead to analytical errors. This study deals with the assessment of the physico– chemicaland bacteriological parameters of water samples from River Amba during the period of August 2017 to January 2018 and developing regression models. Water quality Parameters such as Temperature, Turbidity (NTU), Suspended solids (mg/l), Colour, Total solids, Total dissolved solids, Electrical conductivity (μs/cm), pH, Hardness, Chemical Oxygen Demand, Dissolved Oxygen (DO), and Total Coliform were obtained and compared with water quality standards. The results of the water quality analysis of the study in comparison with drinking water quality standard issued byWorld Health Organization(WHO) and National Agency for Food and Drug Administration Control (NAFDAC) revealed that most of the water quality parameters were not adequate to pronounce the water potable. Hence adequate water treatment processes should be employed to make the water fit for consumption and other domestic uses. Statistical analysis was done, in which the systematic correlation and regressionanalysis showed a significant linear relationship between different pairs of water quality parameters. The highest correlation coefficient between different pairs of parameters obtained is (r = 0.999), resulting from the correlation between TS and SS. Multiple regression analysis was also carried out and regression equations were developed. It was observed that the parameters studied had a positive correlation with each other.
Time, in the globalized world, is one of the most important factors about the economy, science and health. Mankind has made various efforts to use time efficiently for many years. In these studies transport came to the fore and it has become indispensable. In the light of today's technological conditions, air transport is developing at an increasing rate. Every day many aircrafts are produced, which have different speeds, weight and volume, for serve to transport. Therefore to make structures for easy and safe transport need a stable soil. Particularly suitable areas for the airport grounds in cities today, not being physically proper that construction of the airport made on soil with low bearing capacity, swelling potential of an expansive soil, settlement of soil etc, areas. In this study, soil problems encountered in the construction of airports will be explained and a summary of studies on the solution of these problems will be presented.
People in a big city as Antananarivo, capital of Madagascar, have leads to take street foods for their daily nutritional needs. This food habits may be a risk for consumers due to contaminations from street environment and bad practices related to hygiene. This study aimed to examine the quality and safety of street vended foods in Antananarivo, on January 2016 to December 2017.Six hundred and sixty two samples including 126samples of melting salads, 70 beef skewers, 54 chicken skewers, and typical Malagasy foods as : mofoanana (67 samples), mofogasy (64 samples), ramanonaka (64), makasaoka (66), mofoakondro (62) and kobandravina(89);were randomly collected from the streetvendors in Antananarivo marketsto evaluate their bacteriological quality.International Methods (ISO) was adopted for to find the load of Total Aerobic Bacteria andEnterobateriaceae,Escherichia coli and to search pathogen bacteria as Salmonella, Campylobacter jejuni, Escherichia coli O157H7 and Bacillus cereus in these foods.The results revealed that the mean values ofthe Total Aerobic Bacteria count was 0.1x106- 4.8x106cfu/g. Enterobacteriaceaecount range from 0.4x102 to 1.9x102cfu/g. Escherichia coli count range from 0.04x102cfu/g. to 0.19 x102cfu/g.Salmonellawas only present in melting salads, beef skewers and chicken skewers samples. Bacillus cereus count range from 0,1x102 to 1,5x102cfu/g. Campylobacter jejuniwas only present in samples of ramanonaka and kobandravina. Two strains of presumptive Eschercichia coli O157 H7 (βglucuronidase -) were isolated. PCR method was used to confirm the identity of these two isolates. A high contamination above 106 cfu/g food and the presence of potential pathogens bacteria could be hazardous. Systematic inspections and training of food vendors on food hygiene and application of hazard analysis critical control point (HACCP) has been recognised as measures to guarantee improvement of the quality of street foods.
Scored tablets provide dose flexibility, ease of swallowing and cost savings. However, some problems with scored tablets can be confronted like difficulty of breaking, unequally breaking and loss of mass upon breaking. This paper investigates the effect of score lines on the density distribution using continuum modelling. In keeping with previous work in the pharmaceutical field, a modified Drucker Prager Cap model is described briefly and used in the simulations. Coulomb friction is included between powder and tools. The microcrystalline cellulose (MCC) Vivapur® 102 was used to identify the model parameters using experimental tests with instrumented die, shear cell and diametrical crushing. The obtained results indicate that simulations may be useful not only to determine density distributions within tablets, but also may provide indications about performance of score lines.
In a plethora of countries, buildings are adapted to the local climate condition using sustainable architecture techniques and materials, thereby the highest level of climatic comfort is provided. For example, the walls and roofs reflecting sunlight have been used for centuries in the warm regions of the world, while in the cold regions, the maximum use of solar energy has been tended.
The process of modernization has created a high density, thereby demand for fast and affordable constructions in cities has subsequently increased, resulting in reduced attention to environmentally sustainable architecture techniques that, in turn, has led to the financial loss and scarcity of non-renewable energy resources over long periods of time.
Regarding the energy crisis and the necessity of saving non-renewable energy, the reduced need to use heating/cooling systems is assumed to be one of the key goals in advanced building design.
The present study was conducted based on causal research and simulation. Design Builder thermal simulation software was used as the tool to this end. Therefore, a building with/out solar chimney was modeled and analyzed to identify the effect of solar chimney on the amount of energy used for heating.
The control of motor rotation speed by the change of resistor resistance value in armature circuit is called ‘resistor control”. For the regulation of resistance value R0, included in armature winding circuit, we can use various technical solutions. The most used solution is the discrete variation of armature added resistance value by shunting its parts with contactors contacts. Nowadays, the change of resistor resistance in armature circuit can be realized by shunting with a given porosity γ of resistor R0 trough electronic keys. In this paper, we study the design of control system represented on figure 1.
A poultry yield prediction model have then designed using a data mining and machine learning technique called Classification and Regression Tree (CART) algorithm. The developed model has been optimized and pruned using the Reduced Error Pruning (REP) algorithm to improve prediction accuracy. An algorithm to make the prediction model flexible and capable of making predictions irrespective of poultry size or population has been proposed. The model can be used by poultry farmers to predict yield even before a breeding season. The model can also be used to help farmers take decisions to ensure desirable yield at the end of the breeding season.
Today, Web site design is used to make sites useful to users, with accessible functions, resources and information. Therefore, that design involves use of methodologies that allow an adequate structuring of them resources and organization, permitting users to access them quickly, easily and intuitively. This research consisted of a usability study oriented to website structure designers using a methodology based on concepts of ontology design. This study includes a planning to evaluate the design and the structure of website in aspects such as: ease of use, efficient access to information and performance on the tasks focused to total satisfaction of end user. Heuristic tests were used as diagnostic tools to evaluate usability of website design structures; these were supported by a heuristic evaluation guide and in the Sirius methodology[3]. The results obtained from them, allowed us to detect opportunities for improvement and optimization in website design, and in refining the Web interface oriented to end users.
Acceptance of a website is determined by various factors, one of the most important is the organization that allows users to access to functions, resources and information that it contains. This work consisted of a study of comparative usability between a website designed using principles of linguistics and design of ontologies and other using a strategy of a commercial product. A plan was designed and applied to evaluate the following aspects of website: ease of use, efficiency to access its information, efficacy to perform tasks and user satisfaction. Heuristic and user tests were used as diagnostic tools in usability evaluations, and an observation guide was made by an external evaluator as a complement to previous tests. The results clearly shown that is better use the proposed website design methodology. This allows to create site more structured, functional and with greater ease of access to resources that it contain.
Epoxidized sunflower oil (ESO) has been used to toughen epoxy resin GELR 128 cured with an accelerated aliphatic amine curing agent (Kingcure K-11) at room temperature. There was difference in the properties of the polymer composite materials based on epoxy resin GELR 128 cured by Kingcure K-11 between two processes: one-stage process and two-stage process for mixing ESO with epoxy resin GELR 128 at various content of ESO. The results showed that the two-stage process is considered to be more advantageous than the one-stage process. It can be concluded that the impact strength, critical stress intensity factor Kic and decomposition temperature of the polymer composite materials based on epoxy resin GELR 128 cured by Kingcure K-11 with content of ESO 5 phr in two-stage process was greater ones in one-stage process (impact strength: 35.012 kJ/m2, Kic: 2.72 MPa and decomposition temperature: 385.81 0C respectively).
A Network Intrusion Detection System (NIDS) monitors a network for malicious activities or policy violations [1]. The Kernel-based Virtual Machine (KVM) is a full virtualization solution for Linux on x86 hardware virtualization extensions [2]. We design and implement a back-propagation network intrusion detection system in KVM. Compared to traditional Back Propagation (BP) NIDS, the Particle Swarm Optimization (PSO) algorithm is applied to improve efficiency. The results show an improved system in terms of recall and precision along with missing detection rates.
In this paper, we consider the scaling invariant spaces for fractional Navier-Stokes in the
Lebesgue spaces ( ) p n L R and homogeneous Besov spaces
, ( ) s n
p q B R respectively.
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Bioremediation of soils polluted by petroleum hydrocarbons by Pseudomonas putida
1. International Journal of Innovation Engineering and Science Research
www.ijiesr.com
Volume 2 Issue 5 September-October 2018 9|P a g e
ABSTRACT
Bioremediation of soils polluted by petroleum
hydrocarbons by Pseudomonas putida
Tsirinirindravo H.L.1
, Rakotoarisoa M.T .1
, Randrianierenana L.A.1
, Andrianarisoa B1
,
Raherimandimby M.1
, Andriamady H.1
, Randriamiarisoandraibe H.1
,
Andriampenotanjona B. F.1
,Rajaobelinjatovo N.P1
,Raharijaona T.R.2
,DePercin G.3
,
Delandes X.3
, Pierluigi B.4
,Larroque M.5
, Margout D5
.
1. Département de Biochimie fondamentale et appliquée, Faculté des Sciences d’Antananarivo, Madagascar
2. Ecole Supérieure Polytechnique d’Antananarivo, Madagascar
3. Ecole spéciale ESTPI, Paris
4. International University Network on Cultural and BiologicalDiversity (IUNCBD), Italie
5. Faculté de Pharmacie de Montpellier, France
In order to clean up soils contaminated with hydrocarbons, the bioremediation activity of
Pseudomonas putida was studied. Pseudomonas putida is a bacterium that can withstand the
harshest environmental conditions. It is able to metabolize a wide range of petroleum hydrocarbons
which is used as a source of carbon and energy. Given the potential of this microorganism, an
experiment wasconducted on this strain.
For the isolation of this microorganism, a sample ofsoil from the Vakinankaratra region in the urban
commune of Antsirabe II, Madagascar was microbiologically analysed. The bacterial identification was
based on a study of the morphological, physicochemical and sequential analysis of the 16S rDNA
gene.
The isolated strain was then inoculated into soil polluted by diesel engine oil from the garage at
Universityof Saint Joseph Antsirabe, Madagascar. The kinetics ofbacterial growth showed that the
biomass increased from 4.10
7
to 3.10
13
CFU/g at the end of the experimentation. A growth rate of
0,32h
-1
and a generation time of 2,16 hours were noted. The quantification of the residual
hydrocarbons according to the EPA method (Environmental Protection Agency) 3540C have made it
possible to deduce the capacity of degradation of the bacterial strain which is 0,2 mg of hydrocarbon
per gram of soil per day. After 3 months of biological treatment, the concentration of the residual
petroleum hydrocarbons had been reduced by 30%(from 18000 mg/kg to 5000 mg/kg). Thus, the
ability of Pseudomonas putida to decontaminate polluted soils with hydrocarbons has been observed.
Key words: petroleum hydrocarbons, bioremediation, Pseudomonas putida, soil, oil.
I. INTRODUCTION
Petroleum hydrocarbons are potentially toxic organic compounds. Their presence in the soil has
negative impacts on both environmental quality and human health (Robert, 1996). Several technics
have been proposed to reduce the amountof petroleum hydrocarbons in the soil. Among these
technics is bioremediationwhichis to the most ecological and the cheapest.Itconsists to use living
organisms or microorganisms to clean up contaminated sites (Ballerini and Vandecasteele, 1999).
Bacteria are the most used in bioremediation, however, depolluting activities have been seenwith
algae (Selenastrumcapricornutum) and fungi (Aspergillusniger). Moreover, Madagascar has a broader
microbial diversity. There are pathogens microorganisms as Salmonella, Escherichia coli, ….And
there are beneficial microorganisms, it is even profitable for food, and environments because it allows
to protect them, improve their qualities (Tsirinirindravo and al, 2016 ;Mananjara and al, 2016).
2. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 10|P a g e
This is why a study was conducted on the bioremediation the soils polluted with hydrocarbons by
Pseudomonas putida in order to develop an efficient bioremediation system. P. putida is an ubiquitous
bacterium, mostly found in therhizosphere zone ofPinusradiata (Mukerjiet al., 2006). The bacterium is
classified as a chemo-organotroph because of its capability to metabolize a wide range of carbon
compounds including petroleum hydrocarbons. Different manipulations were carried out at the
microbiological and physico-chemical laboratories of the University of Saint Joseph Antsirabe,
Madagascar.
II. MATERIALS AND METHODS
Biological material
The sampling of the soils to analyze was taken from the soil inVakinankaratra region in the
urban commune of Antsirabe II, Madagascar. Pseudomonas putida is a ubiquitous bacterium found in
large quantity in the root zone of Pinusradiata (Bowen et al., 1976). The samples were taken from
three different rhizosphere zones of Pinusradiata at 15 cm deep.
Bacterial isolation
To make the soil suspension, 10 g of the sample of soil sample are mixed with 90 ml of
Buffered Peptone Water. 1 ml of this suspension is plated oncetrimide agar plates and then incubated
at 30°C for 48 hours. The fluorescent bacteria colonies at 254 nm are purified in view of being
identified (Maksimovaet al., 1994).
Bacterial identification
The purified bacterial colonies are identified by a succession of biochemical tests (Garrityet
al., 2002) and 16S rDNA sequence analysis.
Phenotypic study
The identification of the pure strains is firstly based on a macroscopic observation to
determine cultural characteristics. A microscopic observation ata fresh state and after Gram staining
made it possible to characterize the bacterial morphology. The respiratory type as well as the
respiratory enzymes are both determined. The biochemical characterization is carried out through
various tests, namely: test on medium HAJNA-KLIGLER, test on medium SIMMONS CITRATE, test
on medium MANNITOL-MOBILITY-NITRATE, test on medium LYSINE-IRON, gelatintest (Nelson,
2002) and the carbon auxanogram (Latouret al., 1997). In addition, an analysis of the 16S rDNA
sequence of the presumtived strainofP. putida is necessary in order to confirm the strain identity.
III. Molecular study
Extraction of genomic DNA
The "universal" extraction procedure using SDS and proteinase K has been adopted with some
modifications (Daniel et al., 1995). In order to realizethe extraction of the genomic DNA, the strain to
be analyzed is firstly cultivated in nutrient broth for 24 hours at 30 °C. Two ml of the bacterial
suspension are poured in sterilized Eppendorf tubes and centrifuged at 12000 rpm for 10 minutes. In
the obtained pellet are poured 467 μl of the TE buffer solution, 3 μl of 20 mg / ml of proteinase K and
30 μl of 10% SDS. After incubation at 37 °C for 12 hours, the released proteins are precipitated with
400 μl of phenol / chloroform (50:50). The DNA suspended in the aqueous phase is then collected by
centrifugation at 12000 rpm for 10 minutes. To ensure that the DNA samples are free of
contaminants, 4μl of RNase (100mg/ml) are added then incubated for 2 minutes at room temperature.
Finally, the DNA is precipitated by addition of sodium acetate and isopropanol, then collected by
centrifugation at 13000 rpm for 10 min and dissolved in TE buffer. The DNA solutions are finally
stored at -20 °C for amplification (Maloy, 1990).
Qualitative analysis of DNA on agarose gel
To prepare the agarose gel, 30ml of TBE buffer are mixed with 0,3g of agarose. The obtained mixture
is boiled until complete dissolution of the agarose. After having put in place the electrophoresis cell
3. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 11|P a g e
and the comb, the gel is poured ina manner as to obtain a thickness of 3 to 5 mm. After the
solidification of the latter, the comb is removed and the gel is immersedin TBEbuffer as migration
buffer. The first well is loaded with a size marker (FERMENTAS, Lambda DNA / HindIII Marker). In
the other well are poured: 8 μl of DNA sample, 2 μl of loading buffer (BIO LABS Gel Loading Dye).
The manipulation is carried out in a horizontal electrophoresis apparatus (BIO RAD, Mini-Sub Cell
GT) at 100 V for 15 minutes. After the migration of the DNA fragments, the gel is stained with
ethidium bromide.
Amplification and sequencing of the 16S rDNA region
The target gene is amplified by conventional PCR in a programmable thermocycler(PTC-100). The
chosenprimer pair limit the rDNA sequence to be replicated.The latter has been described by
Weisburg in 1991:
27F (forward): (5 'AGAGTTTGATCMTGGCTCAG 3')
1492R (reverse): (5 'TACGGYTACCTTGTTACGACTT 3')
The PCR reaction is carried out in 25 μl reaction volume containing 5 μl of genomic DNA, 1 μl of each
primer used, 0,125 μl of Taq polymerase, 2,5μl of PCR buffer, 2 μl of MgCl2, 0,5μl. OfdNTP. The
thermocycler is programmed as follows: an initial DNA denaturation phase at 94 °C for 5 minutes and
then 35 cycles of 1 minute at 94 ° C for DNA denaturation, 1 minute at 60 ° C for hybridization of
primers and 1,5 minutes for elongation phase. The reaction is finished by increasing the temperature
to 72 ° C for 10 minutes. The amplicons obtained are analyzed by agarose gel electrophoresis.
For sequencing the gene of interest, the PCR products are electrophoresed on an automated Applied
Biosystems 3730xl DNA sequencing apparatus, using 50 cm capillary arrays and a POP-7 polymer.
The results obtained are analyzed on PE-Biosystems version 3.7.
Phylogenetic analysis of sequences
The sequences are compared with those found in GenBank database the National Center for
Biotechnology Information (NCBI) using the BLAST program. Genetic affiliation is evaluated using the
Phylip 3.69 software. The results obtained are then represented in the form of a phylogenetic tree.
Biological test
The biological test sits on 3 months. It is realized in a sealed glass container of about 4500 cm
3
to
avoid contamination from outside. The container is covered by perforated aluminum foil to ensure
aeration (Tarayre 2012, Bidaud 1998). The strain identified as Pseudomonas putida constitutes the
inoculum in the experimental device. The setting of the device is done in three steps:
The preparation of the preculture consists in inoculating 3 handles of the strain in 100 ml of nutritive
broth. The preparation is incubated at 30 ° C for 24 hours.
Then, 500 g of soil (dry weight) are autoclaved at 121 ° C at 1.5 bar for 2 hours. The soil is then dried
at 103 ° C for 24 h and placed in a desiccator until it is cooled. 1.8% of diesel engine oil is added to
the soil (El, 1998). Then, 150g of Pinus sawdust representing 30% of the weight of the soil is used as
texturizer (Bidaud, 1998). Finally, 0.4% of yeast beer autoclaved and grinded which useas a source of
nitrogen are also added in the soil (Tarayre, 2012).
The last step consists to inoculate 100ml of preculture in the previously prepared soil. The device is
placed in the shade at room temperature (23 ° C). Every seven days, the soil is stirred and 20 ml of
distilled water are poured on.
Monitoring bacterial growth
The evolution of the bacterial population is followed by microbiological analysis. To do this, 10 g of the
soil sample are mixed with 90 ml of EPT solution in order to make the soil suspension. A succession
of dilutions till 10
-3
is done from the main suspension. Then, 1 ml of each dilution is plated oncetrimide
agar plates. After incubation for 24 hours at 30°C, the bacterial count was done according to the
ISO7218 standard.
4. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 12|P a g e
Determination of Residual Hydrocarbons (Method 3540C)
The residual hydrocarbon concentration in the soil is also evaluated during the process. The 3540C
method was adopted with some modifications. To do this, the soil is prepared in advance before the
extraction of the hydrocarbon. 10 g of soil to be analysed are dried at 103 ° C for 24 h then placed in a
desiccator until cooling. 5 g of dried soil are placed in an extraction cartridge then immersed with 150
ml of hexane. The extraction lasts 16 hours and the evaporation of the product is collected in a flask
of 250 ml capacity containing pumice stones. The flask is then placed in a rotary evaporator to
evaporate the hexane. The solvent and water residues in the flask are removed at 103 °C for 15
minutes, followed by a cooling in a desiccator. Finally, the volume of the extracted product is
determined. In addition, the concentration of residual hydrocarbon present in the soil expressed in mg
/ kg is specified by the following formula (Center of Expertise in Environmental Analysis of Quebec,
2016):
C =
𝐴𝑋𝑉
𝑄
Thus :
- C : concentration of hydrocarbon in theanalysed sample (mg/kg)
- A : concentration of hydrocarbon in the injected extract (ng/μl)
- V : final volume of the analysed extract (ml)
- Q : dry weight of the analysedsample (g)
IV. RESULTS
Bacterial phenotypic identification
The results of the cultural, morphological, physiological and biochemical studies permit to
characterize the isolated bacterial strains.
Six strains were isolated. They are Gram negative, aerobic, mobile, oxidase positive. Fluorescent
pigment production at 254 nm was observed on all strains (fig.1). Two strains produce floral notes.
The biochemical identification of the strains confirmed the identity of the 6 isolated strains. Of which, 2
were identified as Pseudomonasputida, 2 Pseudomonas aeruginosa, 1 Pseudomonas fluorescens
and 1 Pseudomonas chlororaphis. Table 1 gives the biochemical characteristics of each isolated
bacterial strain.
Figure 1 : Bacterial colonies observed at 254 nm after incubation on cetrimide agar
5. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 13|P a g e
Table 1: Biochemical Characteristics of Isolated Bacterial Strains
Strains
HAJNA-
KLIGLER
MANNITOL-
MOTILITY
LYSINE-
IRON
SIMMONSCITRATE
NITRATEREDUCTASE
GELATINTEST
CARBON
AUXANOGRAM
Glu
Lac
H2S
CO2
MANNITOL
MOTILITY
LDC
LDA
SACCHAROSE
XYLOSE
TREHALOSE
Pseudomonas
aeruginosa
- - - - - + - + + + + + + -
Pseudomonas
chlororaphis
- - - - - + - + + - - + + +
Pseudomonas
fluorescens
- - - + - + - + + - + + + -
Pseudomonas
putida
- - - - - + - + + - - + + -
LDC : Lysine DeCarboxylase LDA : Lysine DeAminase Source : author, 2017
Molecular identification
Agarose gel electrophoresis of the extracted DNA fragments revealed unique and intact bands. The
extract can be used for the amplification step of the 16S rDNA region. Electrophoresis is shown in
Figure 2.
Figure 2 : Agarose gel electrophoresis of the genomic DNA of strains presumed to be Pseudomonas
putida P3 and P4 (the first band represents the size marker)
Electrophoresis of the amplified 16S rDNA fragments showed that the size of the fragments was
around 1500bp. Figure 3 shows Electrophoresis of amplified DNA fragments.
6. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 14|P a g e
Figure 3 : Agarose gel electrophoresis of the amplified 16S rDNA fragments of the presumptive strain
Pseudomonas putida P3 and P4 (the first band represents the size marker)
The phylogenetic tree
The assessment in BLAST of the 16S rDNA sequences of the P3 and P4 strains showed respectively
97.8% and 98% sequence similarity with Pseudomonas putida. This similarity is represented by a
phylogenetic tree in Figures 4 and 5.
Figure 4: Phylogenetic tree based on the 16S rDNA sequences of Pseudomonasputida P3 strain and
related species of BLAST database
7. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 15|P a g e
Figure 5: Phylogenetic tree based on 16S rDNA sequences of Pseudomonasputida P4 strain and
related species of BLAST database
Abundance of microorganisms in the rhizosphere zone of Pinusradiata
Fluorescents Pseudomonas are the most common bacterial species found in the rhizosphere zones
(Garcia et al., 2001). P. aeruginosa, P. putida, P. fluorescens, P. clhororaphisbelong to this kind of
bacteria andthey are called PGPR (Plant Growth Promoting Rhizobacteria). Figure 6 shows the
abundance of Pseudomonas species in the root zone of Pinusradiata.
Figure 6: Bacterial distribution of Pseudomonas species in the root zone of Pinusradiata
8. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 16|P a g e
Biological test
The results show a reduction of hydrocarbon concentration from 18000 mg/kg to 5000 mg/kg in 3
months. The degradation rates is around 30%. Figure 7 shows the evolution of the degradation rate of
the hydrocarbon by Pseudomonas putida.
Figure 7: Degradation rate of hydrocarbon on 3 months
The bacterial population increases considerably from 4.10
7
CFU/g to 3.10
13
CFU/g and the generation
time is 2,16h. During the first weeks, bacteria acclimatizes in the new culture medium. It follows an
acceleration phase and exponential phase of growth. The bacterial growth is inversely proportional to
the degradation of the hydrocarbon (Figure 8).
Figure 8: Evolution of the biomass and the hydrocarbon concentrations during the bioremediation of
polluted soil using Pseudomonas putida.
At the end of the experiment, a production of 1,45.10
12
CFU/g of soil per day was noted. This
increased growth correspond to a degradation capacity of 2,52.10
-4
g of hydrocarbon per gram of soil
per day.
9. Tsirinirindravo H.L. et al. “International Journal of Innovation Engineering and Science Research”
Volume 2 Issue 5 September-October 2018 17|P a g e
V. DISCUSSION
The bacterial isolation of a soil sample from the region of Vakinankaratra made it possible to deduce
the presence of 4 different strains of Pseudomonas in the rhizosphere zone of Pinusradiata. These
strains arePseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas chlororaphisand
Peudomonasputida.
Based on the phenotypic characters, it was noted that all these strains belong to the genus
Pseudomonas (Public Health England, 2015). The comparison of the identification results obtained
with those of Bossisal in 2000 makes it possible to identify the strains P3 and P4 as Pseudomonas
putida. However, phenotypic studies are not sufficient to confirm the identity of the strains. Thus, a
molecular characterization has been carried out. The comparison in NCBI of the 16S rDNA of strains
P3 and P4 respectively displays a similarity of 97,8% and 98% with Pseudomonas putida. Results of
the obtained 16S rDNA sequences show that the P3 and P4 encoded strains belong to the species
Pseudomonas putida.
About the experiment carried out onPseudomonas putida, a bioremediation activity was
observed. Indeed, the decrease in hydrocarbon concentration means that the strain is able to use
petroleum hydrocarbon as the unique carbon and energy source (Jirasripongpun, 2002). This
variation of the hydrocarbon concentration corresponds to a degradation capacity of 0,25 mg of
hydrocarbon per day, which is not insignificant. The work carried out by Vinothini and its collaborators
in 2015 on Pseudomonas putida showed a degradation rate of 98,8%, compared to the values
obtained during the experiment, the latter is not yet optimized. Especially since during the first weeks
of experimentation, the rate of degradation was not considerable (5,5%), a preculture in a medium
containing hydrocarbon is therefore required.
There are several parameters to consider for effective bioremediation. On the one hand, the use of a
consortium shows to be more efficient because of the diversity of catabolic enzymes that each
microorganism used (Ghazaliet al., 2004). Physicochemical characters also affect bioremediation.
Specifically, the addition of texturizer such as sawdust promotes the supply of oxygen as an electron
acceptor during metabolic reactions (Bidaud, 1996). The addition of bio-surfactant, temperature, pH,
and moisture also contribute to the increased ability of the bacteria to degrade (Thwaiteset al., 2007).
VI. CONCLUSION
This experiment shows the presence in soil of a strain able to clean up soil contaminated by
petroleum hydrocarbons. The results obtained reflect the ability of Pseudomonas putida to decrease
to 30% the hydrocarbons concentration in 3 months. Despite the complexity of the chemical
composition of the engine oil, the strain is found to be able to degrade this pollutant and to resist its
toxic activity. The effectiveness of bioremediation depends not only on the biological agent used, but
also on the physicochemical characteristics of the soil to be treated. Nitrogen content, moisture and
aeration of the soil significantly affect this bacterial activity. The biological system used, the
bioaugmentation and biostimulation has been appropriate.
However, several factors such as oxygen supply, pH, and humidity must be optimized to promote the
bioremediation activity of the strain.
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