The document describes a student project to investigate the presence of Pseudomonas bacteria strains from soil samples collected from different locations like oil refineries. The methods section outlines the preparation of nutrient agar medium and nutrient broth, inoculation of soil samples into nutrient broth, and incubation of agar plates streaked with samples from the broth. Observations showed growth of Pseudomonas colonies on the agar plates, indicating the ability of Pseudomonas bacteria to break down toxic compounds in oil refinery soils.
Bioremediating Effect of Glomus Hoi and Pseudomonas Aeruginosa on the Organic...IJEAB
This study analyzed the degrading effect of Glomus hoi and Pseudomonas aeruginosa on the organic content and heavy metals of oil refinery effluent polluted soil using Amaranthus cruentus as the test plant. This study was carried out to determine if agricultural activities can be improved using any or both of the micoorganisms. Eight different treatment layouts were used with three replicates for each level of pollution in the treatment layout. Ninety six (96) pots, each containing three kilograms of soil from both sterilized and unsterilized soil were used for the study. Fifty (50) grams of soil inoculum from propagated Arbuscular mycorrhiza was inoculated to a set of twenty four (24) experimental pots containing both sterilized and unsterilized soil before A. cruentus seedlings were transplanted to them. Another set of twenty four (24) pots containing both sterilized and unsterilized soil were injected with thirty (30) mL of P. aeruginosa inoculum solution before transplanting A. cruentus seedlings to them. The third set of twenty four (24) pots received dual inoculation of both fifty (50) grams of soil inoculum containing G. hoi and thirty (30) mL of P. aeruginosa inoculum solution before A. cruentus were transplanted to them. The residual twenty four (24) pots served as the control. Thereafter, pot preparation was arranged in the screenhouse in a randomized block design. The A. cruentus seedlings were raised in nursery for a period of two weeks before they were transplanted to the pots, seedlings were left for 3 days to overcome transplanting shock before contaminating the soil with refinery effluent at various concentrations of 0%, 2%, 4% and 6% v/w. The seedlings were allowed to grow for eight weeks before the termination of the experiment. The pre planting analysis of soil showed that heavy metals analyses (zinc and iron) of sterilized soil had a lower concentration to the unsterilized. The soil pH ranged from 6.3 to 6.8. It also revealed that organic matter and organic carbon content ranged from 0.8% to 1.3% and 0.4% to 1.7%. However, after the experiment, it was discovered in this study that treatments without any microorganism inoculation in sterilized and unsterilized soil had a higher level of % organic carbon and % organic matter content compared to the other treatments that were inoculated with one or two micro-organisms across all the levels of effluent concentration. Heavy metals of soil in all the soil samples were found to increase as the petrochemical effluent increased in concentration. The results obtained were analyzed using Duncan Multiple Range Test (DMRT) and other descriptive statistics. This study opined that the combined use of G. hoi and P. aeruginosa was more effective in improving the organic contentand the reduce heavy metals of oil refinery effluent polluted soil than when either is used singly.
Assessment of the Coliform Bacterial Load of Some Drinking Water Sources in D...Scientific Review SR
Drinking water samples from 5 sachet water companies, 3 boreholes and 2 taps, collected from different locations of Dutse Metropolis of Jigawa State, Nigeria were analysed for coliform bacterial counts using the Membrane Filtration Technique. All the samples contained some amounts of total coliforms, but mostly within permissible levels. Thirty three percent (33%) of the samples from borehole, 60% from sachet water and 100% from the taps contained faecal coliforms, which indicates contamination. Cultures of the faecal coliforms obtained were morphologically identified using the gram-staining procedure and some series of biochemical tests were carried out in order to identify the organisms. The identified organisms were Escherichia coli (E. coli), Klebsiella sp. and Citrobacter sp. Presence of coliforms above the regulatory set standards indicates contamination and un-safeness of the water for drinking. Presence of organisms such as E. coli, Klebsiella sp. and Citrobacter sp. necessitates improvement in monitoring and water hygiene practices to improve the quality of drinking water in the study area.
Bioremediating Effect of Glomus Hoi and Pseudomonas Aeruginosa on the Organic...IJEAB
This study analyzed the degrading effect of Glomus hoi and Pseudomonas aeruginosa on the organic content and heavy metals of oil refinery effluent polluted soil using Amaranthus cruentus as the test plant. This study was carried out to determine if agricultural activities can be improved using any or both of the micoorganisms. Eight different treatment layouts were used with three replicates for each level of pollution in the treatment layout. Ninety six (96) pots, each containing three kilograms of soil from both sterilized and unsterilized soil were used for the study. Fifty (50) grams of soil inoculum from propagated Arbuscular mycorrhiza was inoculated to a set of twenty four (24) experimental pots containing both sterilized and unsterilized soil before A. cruentus seedlings were transplanted to them. Another set of twenty four (24) pots containing both sterilized and unsterilized soil were injected with thirty (30) mL of P. aeruginosa inoculum solution before transplanting A. cruentus seedlings to them. The third set of twenty four (24) pots received dual inoculation of both fifty (50) grams of soil inoculum containing G. hoi and thirty (30) mL of P. aeruginosa inoculum solution before A. cruentus were transplanted to them. The residual twenty four (24) pots served as the control. Thereafter, pot preparation was arranged in the screenhouse in a randomized block design. The A. cruentus seedlings were raised in nursery for a period of two weeks before they were transplanted to the pots, seedlings were left for 3 days to overcome transplanting shock before contaminating the soil with refinery effluent at various concentrations of 0%, 2%, 4% and 6% v/w. The seedlings were allowed to grow for eight weeks before the termination of the experiment. The pre planting analysis of soil showed that heavy metals analyses (zinc and iron) of sterilized soil had a lower concentration to the unsterilized. The soil pH ranged from 6.3 to 6.8. It also revealed that organic matter and organic carbon content ranged from 0.8% to 1.3% and 0.4% to 1.7%. However, after the experiment, it was discovered in this study that treatments without any microorganism inoculation in sterilized and unsterilized soil had a higher level of % organic carbon and % organic matter content compared to the other treatments that were inoculated with one or two micro-organisms across all the levels of effluent concentration. Heavy metals of soil in all the soil samples were found to increase as the petrochemical effluent increased in concentration. The results obtained were analyzed using Duncan Multiple Range Test (DMRT) and other descriptive statistics. This study opined that the combined use of G. hoi and P. aeruginosa was more effective in improving the organic contentand the reduce heavy metals of oil refinery effluent polluted soil than when either is used singly.
Assessment of the Coliform Bacterial Load of Some Drinking Water Sources in D...Scientific Review SR
Drinking water samples from 5 sachet water companies, 3 boreholes and 2 taps, collected from different locations of Dutse Metropolis of Jigawa State, Nigeria were analysed for coliform bacterial counts using the Membrane Filtration Technique. All the samples contained some amounts of total coliforms, but mostly within permissible levels. Thirty three percent (33%) of the samples from borehole, 60% from sachet water and 100% from the taps contained faecal coliforms, which indicates contamination. Cultures of the faecal coliforms obtained were morphologically identified using the gram-staining procedure and some series of biochemical tests were carried out in order to identify the organisms. The identified organisms were Escherichia coli (E. coli), Klebsiella sp. and Citrobacter sp. Presence of coliforms above the regulatory set standards indicates contamination and un-safeness of the water for drinking. Presence of organisms such as E. coli, Klebsiella sp. and Citrobacter sp. necessitates improvement in monitoring and water hygiene practices to improve the quality of drinking water in the study area.
The acute toxicity of oily drill cuttings against the littoral mangrove periwinkle (T. fuscutas) of the Lagos Lagoon was evaluated in the laboratory bioassay. In this study, the result showed that the acute toxicity of oily drill cuttings based on immobility response of Tympanotonus fuscatus increased with time of exposure. The concentration that caused 50% immobility in the organisms at 24hrs, 48hrs, 72hrs and 96hrs were 3808.80ml/L 660.89ml/L, 302.28ml/L and 102.43ml/L respectively. The median lethal concentration of drill cuttings against T. fuscatcus decreased as the duration of exposure increased. The analysis of variance (ANONA) showed that there was significant difference (p< 0.05) between all the treatments at 24, 48, 72 and 96 hours of exposure. The significance of these result is the need to include bio accumulators such as T.fuscatus in monitoring programmes aimed at establishing the environmental level of such pollutant as oily drill cuttings in aquatic ecosystem.
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.
“SCREENING FOR ANTIBIOTIC PRODUCERS IN SOIL FROM THE BANKS OF SEWER CANALS, AND TESTING THE EFFICACY OF ANTIMICROBIAL COMPOUNDS OBTAINED, AGAINST COLIFORMS”
Virulence Phenotype, Physicochemical Properties and Biofilm Formation of Pseu...IJERA Editor
Potable water piping has been demonstrated to serve as a reservoir for opportunistic pathogens bacteria such as Pseudomonas aeruginosa. In this report, we describe the characterization of P. aeruginosa strains isolated from water intended for human consumption by the presence of virulence factors. These strains expressed their suitability for adhesion and the formation of biofilms on polyethylene (PE). Also In this work, we were able to elucidate the factors intervening in adhesion and biofilm formation by showing the role of the substrate, the environment and bacteria. Strong correlation was observed between physicochemical properties especially the electron donor property and the surface percentage covered by cells. These results indicate that this property plays a crucial role in Pseudomonas aeruginosa adherence on the PE surface. In addition, if no relationship was found between the adhesion results and hydrophobicity, it means that this property was not involved in the adhesion process of Pseudomonas aeruginosa on the PE surface.
Evaluation of Selected Botanical Extracts against Subterranean Termite, Copto...Premier Publishers
Coptoterme formosanus is an economically important agricultural and structural pest of warm and humid regions of the world. The main objective of the study was evaluate seed extracts of Brassica nigra and leaves extracts of Acokantra schimperi, Croton macrostachyus and Rhamnus prinoides against C. formosanus workers under laboratory and semi-field conditions. Treatments were consisted of three concentrations levels (5, 10 and 15 weight of botanical powder (g) per 100 ml volume of water) by three replications. Mortality of termite was counted after 24, 48 and 72 hours exposure for both conditions. The results of all botanical extracts at all concentration levels showed that caused mortality of C. formosanus workers. Complete mortality (100%) of C. formosanus was observed after treatment with 15 w/v B. nigra extract at three time intervals under both laboratory and semi-field conditions. Moreover, A. schimperi at 15 w/v concentration also resulted 100% mortality after 48-72 hours of exposure. Brassica nigra extract showed least LC50 (5.63g/100ml) value than other botanical extracts after 24 hours exposure under laboratory condition. Based on their toxicity status extracts of B. nigra > A. schmperi > R. prinoides > C. macrostachyus leaf extracts.
Biosynthesis of Silver Nanoparticles using Rhizophora mucronata and Ceriops d...BRNSS Publication Hub
To find out the bactericidal properties of biosynthesis silver nanoparticles synthesized with Ceriops decandra (C. decandra) and Rhizophora mucronata (R. mucronata), aqueous leaf extract against the cellulolytic bacteria isolated from gut of Macrotermes convolsionarius a termite species. Further, characterization such as ultraviolet, X-ray diffraction (XRD), Fourier transform infrared (FT-IR), and scanning electron microscopy was analyzed for biosynthesized silver nanoparticles. A total of 16 isolates were collected from gut of termites. Of these, seven bacterial isolates exhibited positive cellulolytic test. The isolated cellulolytic bacterial colonies were subjected to antibacterial assay with synthesized silver nanoparticles of the selected mangrove plants. C. decandra showed highest zone of inhibition (16 mm at the concentration of 150 μg/disc) with TGBS15 and R. mucronata showed highest zone of inhibition (18 mm at the concentration of 150 μg/disc) with TGBS09. The synthesized silver nanoparticles of R. mucronata and C. decandra have maximum absorption at 430 and 400 nm. The XRD data showed 2 θ intense values with various degrees such as 25–30°. The FT-IR results revealed prominent peaks in R. mucronata showed absorption bands at 3444, 1622, 1384, 1071, and 471 cm−1 and C. decandra showed absorption bands at 3606, 3418, 2923, 1069, 474, and 426 cm−1, respectively. The biosynthesis of silver nanoparticles with aqueous leaf extract of R. mucronata provides potential source for cellulolytic bacteria of termites
Presentation: Isolation and Characterization of Bacteria from Tropical SoilsNicole Rivera
The purpose of this experiment is to encounter bacteria from tropical soils that have the capability of producing antibiotic. In order to do so, it was necessary to isolate the bacteria, purify it five times and freeze it at a temperature of -80˚C. After purification, gram staining was performed to classify the bacteria as gram positive or negative. The bacteria were characterized by doing purification of genomic DNA and then, a Polymerase Chain Reaction (PCR) with the primers 16sRNA, 16S_1510R and 16S_8F. An electrophoresis was conducted to see if the PCR process went well. The PCR product was purified and analyzed by using Bioinformatics. Finally, both bacteria’s S15UPRCRISENRE30M01 and S15UPRCRISEAFD30M01A had positive results as antibiotic producers for M.luteus.
This presentation includes;
1.INTRODUCTION to Algae
2.HABITAT of Algae
3.SELECTION OF SOURCE- soil & Water
4.ENRICHMENT OF CULTURE- parameters
5.ISOLATION TECHNIQUES:
Establishing Unialgal culture- 6 types
Establishing Axenic culture- 4 steps
4.ISOLATION OF ALGAE FROM WATER- protocol
5.ISOLATION OF ALGAE FROM SOIL- protocol
6.CONCLUSION AND DISCUSSION
7.Reference books
International Journal of Engineering Research and Applications (IJERA) is an open access online peer reviewed international journal that publishes research and review articles in the fields of Computer Science, Neural Networks, Electrical Engineering, Software Engineering, Information Technology, Mechanical Engineering, Chemical Engineering, Plastic Engineering, Food Technology, Textile Engineering, Nano Technology & science, Power Electronics, Electronics & Communication Engineering, Computational mathematics, Image processing, Civil Engineering, Structural Engineering, Environmental Engineering, VLSI Testing & Low Power VLSI Design etc.
The acute toxicity of oily drill cuttings against the littoral mangrove periwinkle (T. fuscutas) of the Lagos Lagoon was evaluated in the laboratory bioassay. In this study, the result showed that the acute toxicity of oily drill cuttings based on immobility response of Tympanotonus fuscatus increased with time of exposure. The concentration that caused 50% immobility in the organisms at 24hrs, 48hrs, 72hrs and 96hrs were 3808.80ml/L 660.89ml/L, 302.28ml/L and 102.43ml/L respectively. The median lethal concentration of drill cuttings against T. fuscatcus decreased as the duration of exposure increased. The analysis of variance (ANONA) showed that there was significant difference (p< 0.05) between all the treatments at 24, 48, 72 and 96 hours of exposure. The significance of these result is the need to include bio accumulators such as T.fuscatus in monitoring programmes aimed at establishing the environmental level of such pollutant as oily drill cuttings in aquatic ecosystem.
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.
“SCREENING FOR ANTIBIOTIC PRODUCERS IN SOIL FROM THE BANKS OF SEWER CANALS, AND TESTING THE EFFICACY OF ANTIMICROBIAL COMPOUNDS OBTAINED, AGAINST COLIFORMS”
Virulence Phenotype, Physicochemical Properties and Biofilm Formation of Pseu...IJERA Editor
Potable water piping has been demonstrated to serve as a reservoir for opportunistic pathogens bacteria such as Pseudomonas aeruginosa. In this report, we describe the characterization of P. aeruginosa strains isolated from water intended for human consumption by the presence of virulence factors. These strains expressed their suitability for adhesion and the formation of biofilms on polyethylene (PE). Also In this work, we were able to elucidate the factors intervening in adhesion and biofilm formation by showing the role of the substrate, the environment and bacteria. Strong correlation was observed between physicochemical properties especially the electron donor property and the surface percentage covered by cells. These results indicate that this property plays a crucial role in Pseudomonas aeruginosa adherence on the PE surface. In addition, if no relationship was found between the adhesion results and hydrophobicity, it means that this property was not involved in the adhesion process of Pseudomonas aeruginosa on the PE surface.
Evaluation of Selected Botanical Extracts against Subterranean Termite, Copto...Premier Publishers
Coptoterme formosanus is an economically important agricultural and structural pest of warm and humid regions of the world. The main objective of the study was evaluate seed extracts of Brassica nigra and leaves extracts of Acokantra schimperi, Croton macrostachyus and Rhamnus prinoides against C. formosanus workers under laboratory and semi-field conditions. Treatments were consisted of three concentrations levels (5, 10 and 15 weight of botanical powder (g) per 100 ml volume of water) by three replications. Mortality of termite was counted after 24, 48 and 72 hours exposure for both conditions. The results of all botanical extracts at all concentration levels showed that caused mortality of C. formosanus workers. Complete mortality (100%) of C. formosanus was observed after treatment with 15 w/v B. nigra extract at three time intervals under both laboratory and semi-field conditions. Moreover, A. schimperi at 15 w/v concentration also resulted 100% mortality after 48-72 hours of exposure. Brassica nigra extract showed least LC50 (5.63g/100ml) value than other botanical extracts after 24 hours exposure under laboratory condition. Based on their toxicity status extracts of B. nigra > A. schmperi > R. prinoides > C. macrostachyus leaf extracts.
Biosynthesis of Silver Nanoparticles using Rhizophora mucronata and Ceriops d...BRNSS Publication Hub
To find out the bactericidal properties of biosynthesis silver nanoparticles synthesized with Ceriops decandra (C. decandra) and Rhizophora mucronata (R. mucronata), aqueous leaf extract against the cellulolytic bacteria isolated from gut of Macrotermes convolsionarius a termite species. Further, characterization such as ultraviolet, X-ray diffraction (XRD), Fourier transform infrared (FT-IR), and scanning electron microscopy was analyzed for biosynthesized silver nanoparticles. A total of 16 isolates were collected from gut of termites. Of these, seven bacterial isolates exhibited positive cellulolytic test. The isolated cellulolytic bacterial colonies were subjected to antibacterial assay with synthesized silver nanoparticles of the selected mangrove plants. C. decandra showed highest zone of inhibition (16 mm at the concentration of 150 μg/disc) with TGBS15 and R. mucronata showed highest zone of inhibition (18 mm at the concentration of 150 μg/disc) with TGBS09. The synthesized silver nanoparticles of R. mucronata and C. decandra have maximum absorption at 430 and 400 nm. The XRD data showed 2 θ intense values with various degrees such as 25–30°. The FT-IR results revealed prominent peaks in R. mucronata showed absorption bands at 3444, 1622, 1384, 1071, and 471 cm−1 and C. decandra showed absorption bands at 3606, 3418, 2923, 1069, 474, and 426 cm−1, respectively. The biosynthesis of silver nanoparticles with aqueous leaf extract of R. mucronata provides potential source for cellulolytic bacteria of termites
Presentation: Isolation and Characterization of Bacteria from Tropical SoilsNicole Rivera
The purpose of this experiment is to encounter bacteria from tropical soils that have the capability of producing antibiotic. In order to do so, it was necessary to isolate the bacteria, purify it five times and freeze it at a temperature of -80˚C. After purification, gram staining was performed to classify the bacteria as gram positive or negative. The bacteria were characterized by doing purification of genomic DNA and then, a Polymerase Chain Reaction (PCR) with the primers 16sRNA, 16S_1510R and 16S_8F. An electrophoresis was conducted to see if the PCR process went well. The PCR product was purified and analyzed by using Bioinformatics. Finally, both bacteria’s S15UPRCRISENRE30M01 and S15UPRCRISEAFD30M01A had positive results as antibiotic producers for M.luteus.
This presentation includes;
1.INTRODUCTION to Algae
2.HABITAT of Algae
3.SELECTION OF SOURCE- soil & Water
4.ENRICHMENT OF CULTURE- parameters
5.ISOLATION TECHNIQUES:
Establishing Unialgal culture- 6 types
Establishing Axenic culture- 4 steps
4.ISOLATION OF ALGAE FROM WATER- protocol
5.ISOLATION OF ALGAE FROM SOIL- protocol
6.CONCLUSION AND DISCUSSION
7.Reference books
International Journal of Engineering Research and Applications (IJERA) is an open access online peer reviewed international journal that publishes research and review articles in the fields of Computer Science, Neural Networks, Electrical Engineering, Software Engineering, Information Technology, Mechanical Engineering, Chemical Engineering, Plastic Engineering, Food Technology, Textile Engineering, Nano Technology & science, Power Electronics, Electronics & Communication Engineering, Computational mathematics, Image processing, Civil Engineering, Structural Engineering, Environmental Engineering, VLSI Testing & Low Power VLSI Design etc.
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
ABSTRACT- Biosurfactant is a structurally diverse group of surface-active molecule, synthesized by microorganisms. Kocuria rosea and Pseudomonas aeruginosa strains isolated from pesticide contaminated soil, which produces biosurfactant were studied. Curd whey was used as a cheap source of growth medium for biosurfactant production. There was formation of stable emulsions of biosurfactant containing broth with vegetable oil and kerosene. These strains produced a clear zone in oil spreading test, which is an indicative of the good biosurfactant activity. Both the strains produced extra cellular biosurfactant in the culture media and showed good foam stability in the culture medium. Biosurfactant was efficiently extracted from the culture broth by acetone-HCl precipitation. The biosurfactants from the two species, namely Kocuria rosea and Pseudomonas aeruginosa were found to have no effects on germinating seedlings of Glycine max, Pisum sativum and Spinacia oleracea, when treated with 25%, 50%, 75% and 100% with the combination of curd whey in the making of 100ml volume. Curd whey as a control was taken with no surfactant. Our study suggested an efficient use in surfactant aided bioremediation in agricultural land.
Key-words- Biosurfactant, Kerosene, Emulsification, Oil spreading, Kocuria rosea, Pseudomonas aeruginosa, Glycine max, Pisum sativum, Spinacia oleracea
I'm delighted to share the PDFs of lab courses in Microbial Physiology and Microbial Genetics. These comprehensive resources cover essential topics in understanding the intricate workings of microbes at a physiological and genetic level. these PDFs provide a detailed roadmap for our laboratory explorations.
Biodegradation of Profenofos Pesticide by Efficient Bacillus Cereus and Klebs...ijsrd.com
The objective of this study to examine potential for the degradation Profenofos pesticide by the bacteria and finding the optimum conditions of bacteria. The growth of the pesticide degrading bacteria was assessed in Mineral salt broth containing 25mg of pesticide at different level temperature levels (25°C,30°C, 35°C & 40°C) and pH levels ( pH 5, pH 6, pH 7 & pH 8) .The maximum growth rate of bacteria was recorded at 35°C and pH 6. Among the tow bacteria the bacteria Bacillus cereus utilized the pesticides effectively and showed maximum growth. Profenofos pesticide was biological degradable.
Biofertilizers definition, classification, bacterial biofertilizers, mass production of bacterial biofertilizers, prospects and constraints of biofertilizers production in hilly regions of Indian states. Liquid biofertilizers and its uses and advatages
1. “TO INVESTIGATE THE PRESENCE OF
PSEUDOMONAS STRAIN (BACTERIA) FROM SAMPLE
OF DIFFERENT LOCATION.”
NASHIK CAMBRIDGE SCHOOL
(Affiliated to CBSE, Delhi Up to Senior Sec Level)
Nasik
ACADEMIC YEAR: 2012-2013
Project Report submitted to
Nasik Cambridge School
CBSE, Delhi up to Sr. Sec Board
BY
TEJASWINI AHIRE
Under the Guidance of Mr. A. SHANKAR RAO
(PGT Biology)
Dept. of Biology,
Nasik Cambridge School
2. ACKNOWLEDGEMENT
This project report is the result of a solo effort and
combined efforts of my guide, friends and my
family. I would like to express my gratitude to all
those who have helped me during my project work.
I am greatly indebted to Prof. Ramchandran Sir and
Mrs. Bharti Ramchandran Mam (Trustees of NCS).
I take this opportunity to thank our Principal Sir Mr.
C. Somu.
I sincerely thank Mr. A. Shankar Rao (PGT-Biology)
who guided me through the project for helping me
to learn the techniques involved in the project.
I would like to thank all the teachers, non – teaching
staff of NCS for their support and motivation.
3. E- MAIL: nashikcambridge2004@rediffmail.com TELEPHONE NO . : 0253-23776382377639
CERTIFICATE
This is to certify that, Tejaswini Ahire 12th
grade Student of NASHIK
CAMBRIDGE SCHOOL (AFFILIATED TO CBSE BOARD, DELHI)
has workedin dept. of biologyon a project- “To investigate the presence
of pseudomonas strain (bacteria) from sample of different location.”
During this period, she got herself acquainted with some techniques in
biology- sterilization, media preparation, weighing, identification of
colonies, isolation of bacteria, spreading and staining.
Mr. C. SOMU Mr. A.SHANKAR RAO
(Principal Sir) (Internal Examiner)
External Examiner
5. ABBREVIATIONS
NAM: - Nutrient Agar Medium
NB : - Nutrient Broth
oC : - Celsius
PH : - Hydrogen ion concentration
Mo’s :- microorganisms
Ps : - Pseudomonas strain
6. Abstract:
The soil collectedfrom oil refineries, petrol bunks,
mechanicalsheds having many toxic compound/
xenobiotics compounds which inhibits/ kills the soil
microbialflora. Based on recent scientific investigation
some soil micro-organismlike pseudomonas strain
present in soil of oil refineries convert the toxic
compound into non toxic forms. This is called
biotransformation/ detoxification.NB was prepared
under sterilized condition and NB was inoculated with
soil sample of oil refineries to culture pseudomonas
strain in an incubator at 37 oC to 24 hours. From NB 2-3
drops of pseudomonas strain were collected& added on
the middle of NAM plates in front of spirit lamp &
spreaded with sterilized spreader kept for incubation in
incubator for 24 hours at 37 oC . After 24 hrs of
incubation agar medium shows the growth of
pseudomonas strain indicates the biodegradationof
toxic compounds in soil of oil refineries.
7. INTRODUCTION
PSEUDOMONAS
Pseudomonas is a genus of Gram-
negative aerobicgammaproteobacteria,belonging to
the family Pseudomonadaceae . The members of the
genus demonstrate a great deal of metabolicdiversity,
and consequently are able to colonize a wide range of
niches.[3] Their ease of culture in vitro and availabilityof
an increasing number
of Pseudomonas strain genome sequences has made the
genus an excellentfocus for scientific research; the best
studied species include P. aeruginosa in its role as an
opportunistic human pathogen, the plant pathogen P.
syringae, the soil bacterium P. putida, and the plant
growth promoting P. fluorescens.
Characteristics
Members of the genus display the followingdefining
characteristics: [13]
Rod shaped
Gram-negative
One or more polar flagella,providing motility
8. Aerobic
Non–spore forming
positive catalase test
Positive oxidase test.
P. putida has the ability to degrade organic solvents
such as toluene.[32] At least one strain of this bacterium
is able to convertmorphine in aqueous solution into the
stronger and somewhat expensive to manufacture.
Pseudomonas bacteria can be found in soil, marshes,
coastal marine habitats, and plant and animal tissue;
generally, these bacteria can tolerate a variety of
physical conditions.
TOXIC COMPOUNDS PRESENT IN OIL REFINERIES,
GAS STATIONS & MECHANICAL SHEDS
As a result of the increased use of automobiles, the
Demand on gasoline/dieselstations is increasing. In
such
Stations, fuel oil, which is classified as hazardous waste
(Bartha & Bossert 1984) is spilled during transfer and
Servicing operations. The accidentalspillage of
hydrocarbons
On the soil may result in a selective increase in
Hydrocarbon-utilizing microorganisms (Venkateswaran
Et al. 1995; Ferrari et al. 1996). The enhancement or
Reduction will depend upon the chemicalcomposition
of
The contaminating hydrocarbons and on the species of
9. Microorganismpresent within the microbialcommunity
Of the particular ecosystem (Atlas 1995). The
distribution
Of the type and the number of microorganisms at a
Site may help to characterizethe site with respect to the
Concentration and age of the contaminant.
The widespread distribution of members of the genus
Pseudomonas in all hydrocarbon-polluted soils of this
Study as well as of the other investigations confirms
their
Prevalenceand reflects their potential in utilizing these
Hydrocarbon contaminants for growth and thus clean
These polluted sites (Cork & Krueger 1991). Therefore,
The different Pseudomonas spp. of this study were
Evaluated for such potential using the modified method
Of Jacobs et al. (1983) through transforming alkanes to
Their corresponding alcohols. Detectionof alcohol
Formation by microorganisms may be an applicable
Approach for evaluating their activity on the simple
Alkanes. Saadoun (2002) reported that this technique
Can be applied for screening of organisms for their
Ability to degrade hydrocarbons.
Bioremediation& biodegradation
Bioremediationof polluted soils through the
introduction
Of exogenous microbialisolates into the polluted
environment
(Bioaugmentation) has gain increasing interest as an
alternative method
10. To biostimulation in situation where the indigenous
species cannot cope with their environmental
pollutants. However, many factors including predation,
competitionwith indigenous microorganisms, nutrient
availability,physicochemicalparameters and other
environmental factors necessary for growth has been
found to influence the survival of bioaugmented
microbes as wellas their bioremediationefficiencies
Many of the previous studies reported the use of
activated contaminated soils or sludge as inoculants for
bioaugmentation but the fair of transferring more
hazardous contaminants through the activated soil or
sludge is also seen as a concern hence, the report of
possible usage of uncontaminated soil with potential
contaminant degrader as inoculants
11. Materials and Methods
Material Required:
I) Glassware:
1) Beaker
2) Conical flask
3) Petri plates
4) Glass rods
5) Syringes
6) Test tubes
7) Measuring cylinder
8) Spirit lamp
9) Spreader
II) Electronic Devices:
1) Electronic Balancer
2) PH meter
3) Auto clave
4) Hot air oven
12. 5) Incubator
6) Hot plate
III) Chemicals:
1) De ionized water
2) Surgical Spirit
3) Nutrient agar medium
4) Nutrient broth
IV) Other requirement:
1) Cottonplug
13. METHODS
Preparation of Nutrient Agar Medium
Take a 250ml of distilled water in 500ml of beaker
To this add 9.25gm of agar medium
Mix thoroughly by using a sterilized glass rod
Adjust the pH of nutrient agar medium in between 7.2 to
7.4
Transfer the medium into conical flask
Add 3 to 4gm of agar-agar and mix thoroughly by using
sterilized glass rod
14. To liquefy agar-agar, boil the medium with the help of
Bunsen burner or hot plate
Close the conical flask with the help of non adsorbent
cotton plug
Sterilize the NAM in an autoclave at 121o c temperature,
16lbs pressure for 15 minutes
After autoclaving collect the NAM
Pour the NAM into the Petri plates in sterilized
condition and keep at room temperature for 10-15
minutes to solidify the nutrient agar medium.
PREPARATION OF NUTRIENT BROTH
Take a 250ml of distilled water in 500ml of beaker
To this add 9.25gm of agar medium
Mix thoroughly by using a sterilized glass rod
15. Adjust the pH of nutrient agar medium in between 7.2 to
7.4
Transfer the medium into conical flask
Close the conical flask with the help of non adsorbent
cotton plug
Sterilize the NAM in an autoclave at 121o c temperature,
16lbs pressure for 15 minutes
After autoclaving collect the NAM
Spreading
Collect 2-3 drops of the pseudomonas culture and
spread it thoroughly on NAM plates with the help of a
sterilized spreader and incubate it in an incubator in an
inverted position.
Preparationof test sample:
1 gm of soil from different locationis added to 9 ml
distill water.
16. Inoculation:
Test sample is inoculated into NB.
Incubation:
Incubate the NAM plates in an incubator for 24 hrs at
37oc.the growth of pseudomonas in NB is observed by
increase in turbidity of NB.
19. Result:
Tiny colonies of pseudomonas strain is observed on
NAM plates that indicates the biodegradationof toxic
compound in soils collectedfrom oil refineries, petrol
bunks & mechanicalshed.
20. REFERENCES
▪ Comprehensive practical biology class-XII
▪ Ananthanarayana-Microbiology
▪ Prescott-Microbiology
▪ Microbiology-Laboratory Manual – S M Reddy & S
Ram Reddy
▪ Vikas publication-Botany
▪ www.wikipedia.com
21. LEGENDS
▪ Fig.1.1 – Photograph showing growth of mouth
microbial flora in control experiment.
▪ Fig1.2 - Photograph showing zone of inhibition for
different toothpastes.
Fig 1.3 – Observation table showing size of zone of
inhibition.
▪ Fig 1.4 – Graph showing zone of inhibition for
different toothpastes against mouth microbial flora.
22. METHODOLOGIES
AUTOCLAVING
Autoclave is a common and most essential
instrument in every biology laboratory. It is used
for sterilization of media (solid and liquid), glass-
wares and rubber products. It is based on principle
that saturated steam heats an object many times
more efficiently than hot air at the same
temperature. This apparatus, when switched on
generates a saturated steam under pressure and
then brings about sterilization. The increased
pressure results in the elevation of boiling point of
water and produces steam with high temperature.
However it is important to note that it is not
pressure that kills the organisms but the high
temperature of the steam. The boiling point of
water at 15 lbs pressure is 121 ° C.
23. HOT AIR OVEN
Oven is another common instrument of
microbiological laboratories and is used for
sterilizationof glassware. It is based on the
principle whether sterilization is accomplished by
dry heat or hot air. An oven consists of an insulated,
heat proof cabinet which maintains a desired
constant temperature with the help of electric
heating mechanism and a thermostat. Normally, all
the routine glassware is sterilized by keeping at 160
° C for 2 hours.
INCUBATOR
An incubator is very similar to oven in construction.
However, its low thermostat is designed in such a
way to maintain low temperature i.e. below 80 °C. It
is used for incubating, maintaining the cultures at
constant desired temperatures or above the
ambient temperature. Like the oven, an incubator
consists of a heating element at the bottom, a
thermostat, temperature probe and devices for
regulating the temperature. Some incubators are
provided with light arrangements to provide light to