Two varieties of pomegranates named Rabab and Hasibi which are harvesting in Yazd (Iran), have been used. Fruits were peeled and dried up under sunlight. They were washed with sterile distilled water. The dried rinds were grinded into a fine powder by Mixing grinder.
The aqueous extract was prepared by dissolving 1g of dry extract with 20 ml of sterilized distilled water, so the final concentration of extract would be 0.05 g/ml, from this solution other concentration were prepared (0.1-0.2) g/ml. the solutions were shaken for 30 min. The extract was centrifuged (30,000 rpm; 15 min) and the supernatant was Separated. To hydroalcoholic extract, 80 g of the powder was extracted with aqueous methanol (75%). The other two concentrations were prepared from soaking sixfold aqueous methanol (75%) with different amounts of powder. At the end of extraction each extract was passed through the Whatman filter paper No. 1 (Whatman, UK). The methanol was removed from residue with rotatory evaporator (نام دستگاه، سال و .. ).
Agar well diffusion method
This method was used to determine the antibacterial activity of the extracts. For preparation of the inoculum including overnight culture of strains (Staphylococcus aureus(ATCC 29737), methicillin resistance Staphylococcus aureus(MRSA)(ATCC 33591) and
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primarily each isolate was cultured in broth medium and incubated on 37 C. overnight. Suspensions of any isolates were prepared that turbidity equal to 0.5 McFarland standards(OD=0.1). Each bacterium had been swabbed with those suspensions on Muller Hinton agar medium. Four wells were created (6mm in diameter and the distance of 25cm from each other) with sterile cork borer. Each well was poured with 100 μl clear supernatant liquid of any concentration. Plates were placed in room temperature to absorb extracts. They incubated for 24 hours at 37C. their diameter of inhibition zone was measured in
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.
Therefore, acid fat, lactose fermentation, mannitol fermentation were not needed to be performed, because they are selective to a specific thing. As a result, Unknown bacteria “W” was concluded to be gram stain positive, endospore positive, bacilli shape (rod shape), and arranged in chains (strepto-). Test Purpose Reagents Observation Results Gram Stain To determine gram reaction of bacteria.
The Unknown Identification Lab was an experiment that provided the opportunity to apply all the tests that were learned in the semester of lab, to identify the two bacterias that remain unknown. Gram- staining and two other tests will be used to identify the unknowns. This experiment is crucial to the understanding of each test, and can benefit in the ability to identify the characteristics of specific bacteria. Having a clearer understanding of the bacteria can further the research of bacteria for medicine, such as antibiotics. The understanding can also help the development of research in the environment.
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.
Chem 51 LB Experiment 3 Report Scaffold: Bromination of Trans-Cinnamic Acid 1. The goal of this experiment was to perform a halogenation reaction through the addition of two bromides from pyridinium tribromide. This was accomplished by reacting trans-cinnamic acid with pyridinium tribromide. After the reaction took place, melting point analysis was conducted to find out the stereochemistry of the product, which could either be syn-addition, anti-addition, or syn + anti-addition. 2.
By Gram staining alone, it was safe to eliminate the three Gram positive bacteria that could have been assigned: S. epidermidis, M. luteus, and B. megaterium. The second step was to streak plate Unknown #10 to observe its macroscopic
They are arranged individually or form pairs, short chains or clusters of irregular shape. Staphylococci are not demanding on cultivation conditions, but grow best at a temperature of 30-37 °C and neutral pH (Yilmaz, Aydin, 2007). Staphylococci are resistant to dryness and the disinfection and hypertonic solutions of NaCl (up to 12%). Nowadays, there are known 27 species of staphylococci with 14 species found on the skin and mucous membranes of humans. Most staphylococci are absolutely harmless (Flynn, Cohen, 2008).
titel achterkant Voorwoord Samenvatting Table of Contents List of abbreviations 1 1. Introduction 2 1.1 Biobased products 2 1.2 RuBisCO 3 1.3 Isochrysis galbana 4 1.4 Tetraselmis sp 4 2. Methods 5 2.1 Size Exclusion Chromatography 5 2.2 SDS-PAGE 6 2.3 Bradford protein assay 7 2.4 Ion Exchange Chromatography 7 2.5 Soxhlet extraction method 8 2.6 Kjeldahl method 8 3. Materials 9 3.1 Size Exclusion Chromatography 9 3.2 SDS-PAGE 9 3.3 Bradford protein assay 9 3.4 Ion Exchange Chromatography 10 3.5 Soxhlet extraction method 10 3.6 Kjeldahl method 10 3.7 Enzymatic digestion 10 3.8 Used strains 11 3.9 General protein extraction method 11 4.
The second ½ of the organism was used for gram staining. The gram stain method was performed on the unknown organism per lab manual page 42 and two gram stain reactions were identified. Organism B was gram positive cocci in grape like clusters. Because organism B was positive I could eliminate Escherichia coli, Enterobacter aerogenes and Proteus vulgaris because these bacteria would be rod shapes. Organism A was gram negative pink rod shaped and because of gram positive morphology I could eliminate Staphylococcus aureus, Streptococcus lactis and Bacillus subtilis.
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.
The B. Vulgaris samples were approximately 1cm3. They were kept the same size to ensure accurate results. A control test was conducted in distilled water to obtain a result to compare. The ethanol treatments were 40% and 70%. To prepare the solutions a 70% ethanol solution was used to make 40%.
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.
A pomegranate is a delicious red, gelatinous type fruit and it contains many