Isolation of Streptomyces sp
The soil samples were collected from various agricultural areas which had been serially diluted up to 10-5. The spread plate technique was followed and incubated at 28°C for 7 days. The Streptomyces sp isolates were maintained on starch casein agar slants.
Screening for antimicrobial activity by well diffusion method
The Streptomyces isolates were grown in starch casein broth at 28ºC for 10 days and the activity were observed by well diffusion method. The culture filtrate was loaded into the wells which had been inoculated with test organism, S.aureus, K.pneumonia, P.aeruginosa and E.coli and incubated for overnight and measured the zone of inhibition.
Extraction of the secondary metabolites from culture supernatant
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Chitosan was suspended in 1% (v/v) acetic acid to attain a 0.3% (w/v) chitosan solution. Sodium tripolyphosphate (TPP) was dissolved in Millipore water at the concentration of 0.1% (w/v). Chitosan control nanoparticles were obtained by the 1ml of TPP solution was poured drop wise to the 5ml chitosan solution under magnetic stirring at 400rpm for 15 min. The formation of nanoparticles was a result of the interaction between the negative charge of TPP and the positive charge of amino groups of chitosan. The bioactive compound loaded chitosan nanocomposites (BCN) were obtained by the addition of a MIC value of the bioactive compound extract to the chitosan contol nanoparticles before centrifugation and constant stirred for another 45 min. Then the mixture of the solution was treated with sonication for 3min. The nanocomposites were separated by centifugation at 10,000rpm for 40 min. Nanocomposites were frequently rinsed with Millipore water. After centrifugation the chitosan nanocomposites were freeze dried by freeze dry system at -70˚C and used for further analysis (Ali et al., 2010).
Nanoparticles were coated on membrane
The synthesized chitosan loaded nanoparticles (CN) and bioactive compound loaded chitosan nanocomposites (BCN) were coated on 0.4 micron membrane by dipping method. The membrane was placed in synthesized nanoparticles solution and kept in
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The diverse mode of vibrations were identified and consigned to conclude the functional groups in the sample.
Morphology study of the nanoparticles
The morphological characteristics (size and shape) of the nanoparticles were examined using a Scanning Electron Microscope (SEM) machine (Phillips C M 12). The sample was placed on a sample holder followed by coating with a conductive metal such as copper, using a sputter coater. The sample is then scanned with a focused fine beam of electrons (Jores et al., 2004).
Membrane filtration technique
The membrane filtration techniques provide a direct count of total coliforms and faecal coliforms in the water sample. This test was carried out in a flow through membrane filtration system on a CN coated membrane, BCN coated membrane and nanoparticles (NCM) non-coated membrane in a sterile condition. . Bacteria are retained on the surface of the membrane which is placed on a suitable selective medium in a sterile container and incubated at an appropriate temperature. If coliforms and/or faecal coliforms are present in the water sample, characteristic colonies form that can be counted directly. The water sample was collected in a sterile container for further analysis. It is clear that the permeability of nanoparticles coated membrane has slightly reduced contrast to non
Identification of bacteria within Unknown Culture #21 In this experiment, an unknown culture of two different types of bacteria was assigned to each person, a number of tests were performed to isolate and identify these bacterial cells. Based on knowledge from the previous experiments completed in lab, a basic understanding of each type of bacteria was used to create a flow chart that would aid the process of identifying the unknown bacteria within the culture. A gram stain that is performed initially will narrow down the types of tests certain bacteria will and will not respond to. In addition to the gram stain, some of the tests that were used include, a catalase test, an Eosin methylene blue (EMB) agar test, a bile esculin test, and a 6.5% sodium chloride (NaCl) test.
The purpose of this lab report is to employ a myriad of skills, tools and, methods learned throughout this semester to perform the appropriate tests for the identification of the assigned unknown bacteria. Add more background information here!!! The most important tools and techniques used during this identification include aseptic technique, microscopic examination and, the use of selective and differential media. Aseptic technique is an important tool for microbiologists. It is imperative that aseptic technique is maintained throughout the length of any test to avoid any cross-contamination that may lead to inaccurate results.
In this experiment, we cultivated an unknown specimen containing two microorganisms. The purpose of this experiment was to use a variety of biochemical test previously learned in the lab to identify the unknown bacteria. The identification of unknown bacteria is a major part of microbiology. Microbiologist observe samples such as blood and sputum in the laboratory for the presence of microorganisms. Identifying unknown bacteria is extremely important in clinical settings because it helps physicians find treatment for infections.
Unknown Lab Report Unknown # 25 By: Jenna Riordan March 19, 2018 Bio 2843 1. Introduction Microbiology is the study of microorganisms found in all different environments throughout Earth, from the hot thermal vents at the bottom of the ocean to the ice at the top of a mountain.
In the laboratory, identification of an unknown bacterium is often necessary. In the lab, a random sample consisting of three different bacteria was selected. The sample contained one gram-positive, one gram-negative paracolon, and one gram-negative coliform. The purpose of the experiment is to identify each of the three species that the mixture contained. After receiving an unknown mixture, the sample was streaked for isolation onto TSA, blood agar, and MacConkey plates.
More specifically the aim was to investigate what effect 40% and 70% ethanol solutions had on a B. Vulgaris cell membrane and then compare them to the same test with distilled water. It was hypothesised that the ethanol solution would increase the membrane permeability. From the results the hypothesis can be supported. The topic of cell membranes have been extensively researched, meaning that there is no limit to information and sources of information of the subject. The effects of alcohols on membrane have also been researched quite extensively.
The erythromycin resistance gene is carried by Tn4351. erythromycin resistance colonies were transfer to LB agar containing 200µgml-1 thrimethoprim. Non of the colonies could grow in this medium and no free vector (R751) was obtained in plasmid miniprep. This indicates that no replication of R751 occurred. Colony blot hybridization was done separately to discover if Tn4351 and/or R751 had inserted into the chromosome of F. chinesis.
To begin, during this lab experiment, genetic transformation was successfully carried out. After observing the agar plates, it was found that only the plate with ampicillin and no pGLO plasmid did not grow any of the E.coli bacteria. All three of the other plates grew the E.coli bacteria, however it grew differently in each plate. In the control plate where the pGLO plasmid, ampicillin, and arabinose were not present, the bacteria grew in the pattern that it was spread in originally. In the two other plates, bacteria grew in colonies that eventually joined together due to prolonged time in the incubator.
After a gram stain was done unknown #257 was identified as a gram positive organism because when observed under the microscope the organism appeared purple with cocci in clusters. The organism was also catalase positive which means that it produced enzyme catalase and bubbled when hydrogen peroxide was added to it. Three test were conducted based on the result of the gram staining procedure. Blood agar with a Novobiocin disk was chosen as well as DNase (DNA) and Mannitol Salts (MSA) agar. The Blood agar is a bright red, opaque plate and the streaking or the inoculation technique was a modified streaking for isolation with a heavy quadrant one.
More specifically the aim was to investigate what effect 40% and 70% ethanol solutions had on a B. Vulgaris cell membrane and then compare them to the same test with distilled water. It was hypothesised that the ethanol solution would increase the membrane permeability. From the results the hypothesis can be supported. Cell membranes are a core aspect of understanding cells which helps to understand humans and other living creatures. Therefore the topic of cell membranes has been extensively researched, meaning that there is no limit to information and sources of information of the subject.
The sterile cotton swab was inserted in the S. epidermidis culture and twirled around to obtain a specimen. The entire plate was inoculate with the swab from top to bottom, to achieve a lawn of growth. The dry forceps was used to remove the antibiotic disk into the appropriate spot on the plate. This process was repeated for the all antibiotics with aseptic technique being used. The plate was incubate with lid up on the bookshelf at room temperature for 48 hours.
If the consistency or integrity of the membrane is damaged the elements stored in
INTRODUCTION: Arginase is an enzyme- enzymes are biological catalyst which drives a reaction at the speed of life. Arginase is a hydrolase, hydrolases catalyze hydrolysis reactions, this is determined via the E.C number (Nelson and Cox 2008). Arginase has the EC number is 3.5.3.1 (Schomburg 2015). The enzyme ‘commission number’ is the arithmetical classification that is used for enzymes which indicates the chemical reaction they catalyze.
Biochemical tests are the tests used for the identification of bacterial species based on the differences in the biochemical activities of different bacteria. Bacterial physiology differs from one species to the other. These differences in carbohydrate metabolism, protein metabolism, fat metabolism, production of certain enzymes and ability to utilize a particular compound help them to be identified by the biochemical tests. Gram’s stain was originally devised by histologist Hans Christian Gram in 1884. Gram-positive bacteria stain purple, while Gram-negative bacteria stain pink when subjected to Gram staining.
Fix or replace damaged tissue; biology has always been the main concern for scientists. Today, the most important instrument for tissue engineering scientists to produce replacement tissues and implants to repair or replace damaged tissue. Tissue engineering is generating a new field of study in which the principles of engineering and biology to correct the damaged tissue, uses and can renewal, operation and maintenance of tissue healing. In order to use an ideal scaffold Tissue engineering should have features such as non-toxic Cell and tissue properties to be fit.