Enzymatic Ability of Bacteria Experiment
Project 9: Enzymatic Ability of Bacteria ________________ Aim: To determine the metabolic properties of Escherichia coli, Proteus vulgaris, Staphylococcus aureus and Bacillus subtilis. Introduction Metabolic differences are used for the taxonomy of bacteria as bacteria divide asexually, as each bacterial species exhibit metabolic differences on a variety of substrates. A bacteria’s ability to metabolise a substrate depends on the microorganism having the necessary enzymes to metabolise the substrate. It can also be dependant of the microorganism’s ability to transport the substrate into the cell, as some substrates are transported into the cell, then metabolised by intracellular enzymes. Other substrates are metabolised by extracellular enzymes (enzymes are secreted from the cell which break down the substrates to smaller molecules that are transported onto the cell then metabolised further by intracellular enyzmes). Method Three different tests were conducted to investigate the metabolic properties of Escherichia coli, Proteus vulgaris, Staphylococcus aureus and Bacillus subtilis. For full details of the tests conducted and the theory behind them, please refer to Maddox et al (2012) Fermentation Test This test is designed to see the ability of the microorganism to metabolise carbohydrates. E.coli, P.vulgarus, S.aureus and B.subtilis were inoculated in three
Development of salmonella vaccine
DEVELOPMENT OF SALMONELLA VACCINE Abstract There are a number of specific characteristics to be considered when developing and manufacturing vaccines. Special characteristics for these vaccines comprise: . The appropriate level of attenuation, the balance between safety and immunogenicity, 2. The genetic stability of the organisms combined with environmental risk assessment, 3. The challenge of old-fashioned upstream and downstream methods in combination with quality control of the final product, and 4. The release requirements. Vaccination has proven to be the most efficient, cost-effective means for preventing a wide variety of infectious diseases. Vaccine development and manufacturing, however, poses several challenges. Inherent to all biological systems is the difficulty to achieve robust production processes guaranteeing reproducible efficacy and safety of the products. Highly automated bioprocess systems or advanced analytical systems used for closed-loop control may be a solution to overcome this. Yet these techniques are still not standard in the industrial environment or cannot be applied to vaccine production processes because of historical reasons, as some of the vaccines have been developed early in the previous century. Because vaccines are administered to healthy people, the efficacy and safety of vaccines are most important. Regulatory GMP (Good
nasal carriage of s.aureus
Analytical microbiology lab report Name : Apu sarwar Class : analytical and pharmaceutical chemistry Date of practical: 2/10/12 and 9/10/12 and 16/10/12 ________________ Title: Estimation of the prevalence of nasal carriage of Staphylococcus aureus in the nasal passages of this class of college students.(three part all together) Objective: This prospective investigation on healthy college student is undertaking to estimate the prevalence of S.aureus nasal carriage and identify the micro organism by using different media and test like gram stain. Introduction: Staphylococci are often found in the human nasal cavity (and on other mucous membranes) as well as on the skin. S. aureus is with some frequency found as normal human flora, Approximately 30% of adults and most children are healthy periodic nasopharyngeal carriers of S. aureus. Staphylococcus aureus (coagulate-positive staphylococci). Staphylococcus aureus is the most pathogenic species and is implicated in a variety of infections. to diagnostic them it is very important to identify this microorganism. there is many way the micro organism can identify. Here different media and staining will apply to identify the organism. Method: for material and method the lab manual was followed. Result : Table 1 : Identification of organisms from the nasal swab in class 1. Identification technique appearance Results
An investigation to show the effects of temperature on the rate of diffusion
An investigation to show the effects of temperature on the rate of diffusion Aim: This experiment aims to determine what effects temperature has on the rate of diffusion on the plasma membrane of a beetroot sample. Hypothesis: Evidence found from earlier research has shown that as temperature increases, the plasma membrane breaks which causes the cytoplasm, and other substances contained within the membrane to leak out. This causes the colour pigment to be released. The rate at which the colour pigment is dependent on the temperature of the solute for example higher temperature means diffusion occurs more quickly. Background Research: Substances are exchanged between cells and their surroundings, across cell surface membrane. Cells obtain their requirements to function by allowing molecules to move across membranes through methods such as diffusion. Diffusion is the process by which a substance moves from a region of high concentration, to a low concentration of the same substance. Diffusion can also be defined as the net movement of molecules or ions from a region of high concentration to an area of low concentration through a partially permeable membrane. The molecules move down a concentration gradient. The rate of diffusion is affected by several factors: } The concentration gradient of the substance effects the rate of diffusion. If the concentration is greater
Use of selective and differential media to identify bacteria
Introduction MacConkey agar is generally used in the isolation of gram-negative bacteria and the differentiation of lactose fermenting from lactose non-fermenting gram-negative bacteria. MacConkey agar is also common to differentiate bacteria by their characteristics to ferment sugars other than lactose such as lactose is being replaced in the medium by another sugar. These modified media are widely used to differentiate gram-negative bacteria or to differentiate between phenotypes with mutations (Vasanthakumari, 2009). MacConkey can be either selective or differential media to be used in the isolation and differentiation of gram negative rods. The inclusion of crystal violet and bile salts in the media prevent the growth of gram-positive bacteria and fastidious gram-negative bacteria . Some gram negative bacteria are able to tolerate bile due to their relatively bile-resistant outer membrane which protects the bile-sensitive cytoplasmic membrane. Gram negative bacteria growing on the media are differentiated by their characteristic to ferment sugar lactose. Bacteria that can ferment lactose causes the pH of the MacConkey media to drop and the change in pH is detected to be red in colour. As the pH continues to drop in the media, the neutral red is then absorbed by the bacteria which appears as bright pink to red colonies on the agar (Pattison et al., 2008). Gram-negative
Antigen-Antibody Interactions: an analysis
Antigen-Antibody Interactions: an analysis Introduction Common to all antibodies is the specificity they exhibit in binding to epitopes present on the surface of antigen in which ultimately a lattice formation results due to cross-linking. The present experiment exploits this interaction and is physically observed as agglutination (when antigen is present on a cell) or precipitation (antigen free-floating in solution) (Elek et al., 1964). Presently, the experiment uses both qualitative and quantitative analyses in order to ascertain the levels of antibodies in sera; such information, for example, is useful in assessing the presence of a current infection in a patient. More specifically, during Part A, presence of agglutination, due to the interaction between Salmonella H (flagellar) and O (somatic) antigens and the appropriate antibodies, was used as a qualitative measure in order to assess whether the patient is currently infected with Salmonella typhi or paratyphi A strains, a process known as the Widal test (Parry et al., 1999). Part B employed a quantitative analysis to determine the level of precipitation upon interaction between bovine serum albumin (BSA) antigen and antiBSA to allow the determination of the concentration of antibody in an original antiserum sample. Materials and Method As per BIOL3141 Infection and Immunity Laboratory Manual 2011 pg 19 -26.
Enzymes - Competitive inhibition.
Enzymes Most of these interactions are weak and especially so if the atoms involved are farther than about one angstrom from each other. So successful binding of enzyme and substrate requires that the two molecules be able to approach each other closely over a fairly broad surface. Thus the analogy that a substrate molecule binds its enzyme like a key in a lock. This requirement for complementarity in the configuration of substrate and enzyme explains the remarkable specificity of most enzymes. Generally, a given enzyme is able to catalyze only a single chemical reaction or, at most, a few reactions involving substrates sharing the same general structure. Competitive inhibition The necessity for a close, if brief, fit between enzyme and substrate explains the phenomenon of competitive inhibition. One of the enzymes needed for the release of energy within the cell is succinic dehydrogenase. Link to illustrated discussion of the citric acid cycle. It catalyzes the oxidation (by the removal of two hydrogen atoms) of succinic acid (a). If one adds malonic acid to cells, or to a test tube mixture of succinic acid and the enzyme, the action of the enzyme is strongly inhibited. This is because the structure of malonic acid allows it to bind to the same site on the enzyme (b). But there is no oxidation so no speedy release of products. The inhibition is called
Mining process of copper.
Figure 1 - The reaction which takes place when the ligand is dissolved in an organic solvent, then shaken with the leaching solution Figure 2 - Simple flow chart of a typical leaching operation Conventionally, copper was mined using the method of smelting. This way, the copper ore is mined, crushed, ground, concentrated, smelted and refined. In practice, only high-grade ores were considered as this created maximum profit. This process has it's draw-backs though. The ore has to be crushed extremely finely - to talcum powder thickness - before the process can start. Also, the ore must contain copper minerals in sulphide form (such as chalcolite or Covellite). The major draw- back was the huge amount of Sulphur Dioxide produced usually more than the copper itself. During the 1980's, a process revolutionised the way copper was mined. Commonly known as the leach- solvent extraction-electrowinning process (SX/EW), it allowed copper to be extracted from low- grade ores and mine wastes. The process was incedentally discovered when a blue-green solution was discovered near mine wastes. On investigation, although aberrant, the solution occured naturally through the work of micro-organisms. There were two types: Thiobacillus ferro-oxidans - which oxide the Fe2+ ions; and thiobacillus thio-oxidans - the S2-. Biotechnologists soon discovered that spraying the
Tiny Bubbles Biology Lab
Tiny Bubbles Biology Lab Purpose: The purpose of this lab is to design and conduct an experiment that explores the effect of temperature as a factor on the rate of the breakdown of hydrogen peroxide by yeast. Materials: * 500 mL of Hydrogen peroxide * Yeast suspension * 50 felt disks * 5 test tubes * Test tube stand * Glass marker * Ice cubes * Tap water * Hotplate * 2 beakers * Temperature probe * Laptop * Connecting wires * Stopwatch * Tongs * 1 graduated cylinder * Test tube brush * Paper and pencil Procedure: Set up: . Connect the temperature probe to the laptop. After putting on the laptop, open the program called LoggerPro. 2. Put on the hotplate and set it to power level 5. 3. Fill one beaker with 500 mL of tap water and set it on the hotplate. 4. With the glass marker, draw a horizontal line at the 3/4 mark on each test tube and place them in the test tube stand. 5. Pour 10 mL of hydrogen peroxide into the graduated cylinder. 6. Pour the hydrogen peroxide in the graduated cylinder into a test tube. 7. Repeat steps 5 and 6 to pour hydrogen peroxide in the four remaining test tubes. 8. Fill the test tubes with tap water till the 3/4 mark and place them back in the stand. Experimental process: Part 1: . Using a tong, soak a felt disk in the yeast solution for 3 seconds. 2. While one person is in control of the felt, another lab
Bacterial Metabolism and Enzymatic Growth
Bacterial Metabolism and Enzymatic Growth Introduction Bacteria differ in the substances from which they derive energy and the enzymes that they produce. Enzymes are biological catalysts; they are substances produced by the body to control the rate of reactions within the body. Particular enzymes, exoenzymes, are produced in large quantities and are excreted into the cell's environment where they speed up the catabolism of insoluble polymers for nutrients (Madigan, 2009). Endoenzymes, unlike exoenzymes are produced in small quantities and remain within the cell (Kocholaty et al, 1938). The purpose of these experiments is to determine the enzymatic and metabolic activities of a variety of bacteria in varying environments. The environments in which the bacteria are observed contain the presence of indole, urease, hydrogen sulfide, and carbohydrates lactose, sucrose, and glucose. The species of tester bacteria cultures for metabolic activities are Escherichia coli (Ec), Pseudomonas aeruoginosa (Pa), Bacillus subtilis (Bs), and Proteus vulgaris (Pv). The enzymatic activities of bacteria are observed in the presence of the enzymes amylase, lipase, and protease. The bacteria used are Escherichia coli, Pseudomonas aeruoginosa, and Bacillus subtilis. The metabolism of the aforementioned carbohydrates results in a change of acidity, evident by a colour change in the presence of the