KOH test
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The KOH test is used to identify Gram-negative and Gram-positive bacteria as an alternative to the Gram stain. It works by dissolving the thin peptidoglycan cell wall of Gram-negative bacteria when exposed to 3% KOH, lysing the cell and releasing viscous DNA strands. Gram-positive bacteria are not lysed by KOH and do not release DNA. To perform the test, a bacterial sample is mixed with 3% KOH on a slide. Gram-negative bacteria will appear viscous and form mucoid strings within 15 seconds, while Gram-positive bacteria will not become viscous or form strings.
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The document discusses laboratory diagnosis of common fungal diseases. It provides data on clinical specimens submitted for fungal isolation from 2004-2006, with the most common being respiratory and body fluids. The most frequent fungal isolates were Candida albicans, C. tropicalis and C. parapsilosis. Methods for diagnosing mycoses include direct microscopic examination, culture techniques, biochemical tests and special tests. Common superficial cutaneous fungal infections affecting the skin, hair and nails are also described, along with their characteristic lesions, causative organisms and laboratory identification.
Synovial fluid examination
Synovial fluid is produced by synovial membranes in joint cavities and acts as a lubricant and shock absorber. It contains hyaluronic acid and filters interstitial fluid. During a joint aspiration procedure, fluid is extracted from a joint through a needle for analysis. Abnormal fluid characteristics can indicate conditions like gout, arthritis, or infection. Precautions are taken to prevent complications, and patients are monitored after the procedure. Laboratory tests of synovial fluid include examining appearance, cell count, crystals, and culturing for bacteria.
Cns infections
There are 4 main routes through which infectious agents can enter the central nervous system: 1) hematogenous spread, 2) direct implantation, 3) local extension from adjacent infections, and 4) spread from the peripheral nervous system. Meningitis refers to inflammation of the meninges and cerebrospinal fluid, while meningoencephalitis refers to inflammation of both the meninges and brain parenchyma. Meningitis is often classified based on the type of inflammatory exudate in the cerebrospinal fluid, such as acute pyogenic (bacterial), aseptic (viral), chronic, etc. The most common causes of acute pyogenic meningitis vary with patient age.
Synovial fluid analysis
one of the tutorial classes during my stint as a teacher in Orthopedics at Manipal University , MMMC , Melaka Malaysia
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The document discusses antibiotics, including their sources, roles, classification, and mechanisms of action. It focuses on several classes of antibiotics that act by inhibiting bacterial cell wall synthesis or protein synthesis. It describes how penicillins and cephalosporins inhibit the final stage of peptidoglycan synthesis in bacterial cell walls. It also discusses how other drugs like glycopeptides, fosfomycins, and aminoglycosides act on bacterial cell components and physiological processes. The classification, mechanisms of action and spectra of several classes of protein synthesis inhibitors are outlined as well.
Biochemical reactions
The document provides information on various biochemical tests performed to identify bacteria, beginning with tests for enzymes. It describes the catalase test in detail, including the principle, reagents, controls, and methods. It notes that catalase is present in most aerobic and facultative anaerobic bacteria except streptococcus species. The document then briefly summarizes several other enzyme tests including oxidase, urease, ONPG, and nitrate reduction. It concludes by stating the order of typical biochemical tests is first enzymes, then tests for specific breakdown products, ability to utilize substances, and metabolism of carbohydrates, proteins and amino acids.
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41. laboratory diagnosis of common fungal diseases
41. laboratory diagnosis of common fungal diseases
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Gram Positive Bacteria
The document discusses gram positive and gram negative bacteria. Specifically, it describes performing biochemical tests on an unknown bacterium that was identified as gram positive. The biochemical tests that were positive included catalase and growth on blood agar, indicating the presence of catalase enzyme and hemolytic activity. Further testing identified the bacterium as Streptococcus pneumoniae. S. pneumoniae is known to cause pneumonia through inhalation and symptoms include cough, fever, and chest pain. Treatment involves antibiotics due to increasing antibiotic resistance in S. pneumoniae.
Gram stain by manoj
Hans Christian Gram, a Danish physician and bacteriologist, developed the Gram staining technique in 1884 to classify bacteria. The technique involves staining a smear of bacteria with crystal violet dye, washing with iodine to form a crystal violet-iodine complex within the cell wall, then decolorizing with alcohol or acetone. Gram-positive bacteria retain the crystal violet due to their thick peptidoglycan layer, appearing dark purple under the microscope. Gram-negative bacteria lose the crystal violet during decolorization due to their thin peptidoglycan layer and outer membrane, appearing red with the counterstain. The Gram stain technique is a simple yet effective way to rapidly classify bacteria and guide
Gram staining miceobiology student
The Gram stain is an important differential staining technique used to classify bacteria into two groups: Gram positive and Gram negative. Gram positive bacteria appear violet after the stain due to their thick peptidoglycan layer trapping the crystal violet dye. Gram negative bacteria appear red as their thin peptidoglycan layer washes away the dye during decolorization, leaving the safranin counterstain visible. The Gram stain procedure involves smearing a bacterial sample, staining with crystal violet and iodine, decolorizing with alcohol or acetone, and counterstaining with safranin.
Senior Microbiology Poster
This document summarizes an experiment that tested the ability of Streptomyces thermocarboxydus bacteria to reduce hexavalent chromium (CrVI). S. thermocarboxydus was isolated from plant roots and identified using 16sRNA sequencing. The experiment involved three parts: 1) inoculating agar plates and measuring bacterial growth with different concentrations of potassium dichromate, 2) inoculating broth tubes with varying dichromate concentrations and measuring absorbance over time, 3) adding dichromate to bacterial supernatant and measuring absorbance. Results showed that S. thermocarboxydus can reduce CrVI to less toxic CrIII, with its secretions and when growing in concentrations up to 0.5 g/L
Staining
Staining techniques allow cells and cell components to be better visualized under a microscope. Simple staining techniques like positive and negative staining make entire cells visible. Differential staining, like Gram staining, selectively stains different types of cells different colors to identify bacteria. Special stains identify specific cell structures like flagella, capsules, or endospores. Staining procedures first involve making a smear, then applying stains followed by washes and examination under a microscope.
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This document provides instructions for performing a Gram stain procedure. The Gram stain allows bacteria to be classified as either Gram-positive or Gram-negative based on differences in their cell wall structure and how they interact with stain reagents. The procedure involves staining a bacterial smear with crystal violet, applying Gram's iodine as a mordant, decolorizing with ethanol or acetone, and counterstaining with safranin. Gram-positive bacteria appear purple or violet while Gram-negative bacteria appear pink or red. The document provides step-by-step instructions for performing the Gram stain and interpreting results under the microscope.
Identification of gram positive and gram negative bacteria
The document describes procedures for identifying gram positive and gram negative bacteria through gram staining and biochemical tests. Gram staining involves staining bacterial smears with crystal violet, iodine, decolorizer, and safranin to identify bacteria as gram positive (purple) or gram negative (pink) based on cell wall structure. Key biochemical tests described include catalase, coagulase, oxidase, indole, and citrate tests, which help identify bacterial species based on enzymatic reactions.
Identification of gram positive and gram negative bacteria
The document describes procedures for identifying gram positive and gram negative bacteria through gram staining and biochemical tests. Gram staining involves staining bacterial smears with crystal violet, iodine, decolorizer, and safranin to identify bacteria as gram positive (purple) or gram negative (pink) based on cell wall structure. Biochemical tests described include catalase, coagulase, oxidase, indole, and citrate tests to further differentiate bacterial species based on enzymatic activity.
Identification of gram positive and gram negative bacteria
The document describes procedures for identifying gram positive and gram negative bacteria through gram staining and biochemical tests. Gram staining involves staining bacterial smears with crystal violet, iodine, decolorizer, and safranin to identify bacteria as gram positive (purple) or gram negative (pink) based on cell wall structure. Biochemical tests described include catalase, coagulase, oxidase, indole, and citrate tests to further differentiate bacterial species based on enzymatic activity.
bacterial different staining technique
This document describes bacterial staining techniques including Gram staining and acid-fast staining. Gram staining differentiates bacteria based on cell wall structure and stains Gram-positive bacteria purple and Gram-negative bacteria pink or red. Acid-fast staining identifies acid-fast bacteria that retain the primary stain carbolfuchsin despite decolorization. Procedures for both Gram staining and acid-fast staining of mixed bacterial samples are provided along with questions about the techniques.
Staining of Bacteria
This document provides information about dyes, stains, and staining methods used in microscopy. It begins with definitions of stains and dyes, explaining that stains are used to facilitate examination under a microscope while dyes are compounds that impart color. It then classifies biological stains as acidic, basic, neutral, metachromatic, or fluorescent based on their chemical properties. Various staining techniques are described including simple staining, negative staining, Gram staining, and acid-fast staining. Specific staining methods and the mechanisms of how they work are explained for simple staining, Gram staining, and acid-fast staining.
Staining
This document provides information about dyes, stains, and staining methods used in microscopy. It begins with definitions of stains and dyes, explaining that stains are used to facilitate examination under a microscope while dyes are compounds that impart color. It then classifies biological stains as acidic, basic, neutral, metachromatic, or fluorescent based on their chemical properties. Various staining techniques are described including simple staining, negative staining, Gram staining, and acid-fast staining. Specific staining methods and the mechanisms of how they work are explained for simple staining, Gram staining, and acid-fast staining.
INTRODUCTION TO MICRO LAB, STAINING TECHNIQUES & MORPHOLOGY OF BACTERIA
This PPT is helpful to understand first practical to 2nd year MBBS student.
I have added 2 video in this PPT to understand staining techniques properly.
Reference: 1 Gram stain video: Dr.G Bhanu prakash animated medical videos
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Gram, Gram And C. Gram
This lab report discusses performing Gram staining on bacterial samples. Gram staining allows bacteria to be classified as either Gram-positive or Gram-negative based on differences in their cell wall structure and how they retain or release dye. The student found that Bacillus subtilis was Gram-positive, retaining the primary dye crystal violet and appearing purple under the microscope. Escherichia coli was found to be Gram-negative as it did not retain the crystal violet dye and appeared pink after the counterstain was applied. The results showed the Gram staining technique was effective at distinguishing between the two bacterial specimens.
Simple and Gram's staining
The document discusses simple staining and differential staining techniques. It explains that simple staining uses a single stain, such as methylene blue or crystal violet, to uniformly stain bacterial cells and allow observation of cell morphology. Differential staining uses multiple stains, like in Gram staining, to distinguish between types of bacteria or cellular structures. The Gram staining procedure is described in detail, involving staining with crystal violet, iodine, decolorization with alcohol, and counterstaining with safranin to differentiate Gram positive from Gram negative bacteria based on differences in cell wall structure. Differential staining techniques provide important information for pathogen identification and selection of appropriate culture media.
Observation Of Water In Sycamore Creek
Based on the results provided in the lab report, the bacterium in Sample 70A is Bacillus cereus. The key results that lead to this conclusion are:
1. The bacterium was Gram-positive rods based on the Gram stain.
2. It formed endospores based on the positive result in the endospore test.
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4. The oxidase test results were unclear but seemed positive.
5. The DNase test was positive, confirming it as B. cereus rather than B. subtilis.
So
Gram stain viva 1
The document discusses the Gram staining technique, which was invented by Hans Christian Gram in 1884. It is a differential staining method that divides bacteria into two groups - Gram positive and Gram negative - based on differences in their cell wall composition that are revealed through a multi-step staining process using dyes. The Gram stain results provide important initial clues about an organism that can guide further diagnostic tests and treatment decisions.
Ex 3-7 The Gram StainPages 195-202Today we will.docx
The document provides instructions for a microbiology lab on observing bacterial cultures, Gram staining, and microscopy. The key points are:
1. Students will observe cultures grown from the previous lab, Gram stain reference organisms, and observe plates under a dissecting microscope.
2. A Gram stain involves staining a smear with crystal violet and safranin, then observing bacteria as either Gram positive or Gram negative based on retention or loss of the primary stain.
3. When using the light microscope, students will draw and label representative samples of bacteria, noting shape, arrangement, staining method, and total magnification.
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The document discusses microbiology and various microbiological techniques. It describes the gram staining procedure used to differentiate bacteria based on cell wall structure. It explains that staphylococcus can be identified using catalase and hemolytic tests on blood agar. The catalase test introduces hydrogen peroxide and observes bubble formation to determine if the bacteria produces the catalase enzyme.
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KOH test
- 1. KOH test Purpose For identification of Gram (-) bacteria. This test replaces the Gram stain. It is much quicker and easier to perform than the traditional Gram stain. It is just as accurate. It is a secondary test for identification of Gram (+) vs. Gram (-) cultures. 3% KOH dissolves cell walls of Gram (-) (thin layer of peptidoglycan), but does not affectGram (+) cell walls. Dissolution of Gram (-) cell wall lyses the cell and spills its contents,including the DNA. DNA is very viscous, and with a large enough cell mass, the DNAstrands can be seen sticking to / dragging from a loop when touched. Gram (+) cells are notlysed, no free DNA, no viscosity will be observed.This test requires a large amount of cells (visible clump). Scrape samples of E. coli,Staphylococcus epidermidis, and your unknown from a solid medium (agar slants). Materials Microscope slide Toothpick 3% KOH Instructions 1. Place one drop of 3% KOH on a microscope slide 2. With a toothpick, transfer a generous amount of bacteria to the drop of KOH. 3. Stir 4. Raise and lower toothpick off slide Expected results KOH + (Gram -); bacteria will be viscous and form a mucoid string in 15 sec. KOH - (Gram +); bacteria do not become viscous or form mucoid strings in 15 sec. Notes Use 24-48h cultures It is a good idea to use + and - controls KOH should be fresh. Discard any bottles with a white precipitate. Coryneform bacteria can be Gram variable as cultures age. Prepared by: Mr. Say Sinith
- 2. 3% KOH test Prepared 3% KOH reagent Place one drop of 3% KOH on slide Pick colony from pure culture Mix and stir organism with the reagent 3% KOH Positive 3% KOH Negative Observed result when raise the loop off slide