Podcast Beta
Questions and Answers
Which of the following genera includes some common human pathogenic species of Enterobacteriaceae?
Salmonella enterica is a genus of the Enterobacteriaceae family known for causing intestinal diseases.
True
Name one differential media used for detecting fecal coliform bacteria.
Eosin Methylene Blue Agar
The enteric bacterium _____ is responsible for causing plague.
Signup and view all the answers
Match the following Enterobacteriaceae species with their characteristics:
Signup and view all the answers
What is the primary purpose of culturing bacteria?
Signup and view all the answers
The pour plate method is primarily used for separating mixed cultures.
Signup and view all the answers
What is a pure culture?
Signup and view all the answers
The _______ method is effective for isolating pure colonies from mixed cultures.
Signup and view all the answers
Match the culturing methods with their primary uses:
Signup and view all the answers
Which of the following is NOT a type of culture medium?
Signup and view all the answers
All bacteria can grow on the same type of culture medium.
Signup and view all the answers
What is one condition that bacteria may require for growth?
Signup and view all the answers
What type of bacteria does Mannitol Salt Agar primarily isolate?
Signup and view all the answers
Lactose fermenters on MacConkey agar produce red colonies.
Signup and view all the answers
What is the primary purpose of Hektoen Enteric Agar?
Signup and view all the answers
The ______ agar is used to differentiate blood hemolysis types.
Signup and view all the answers
Match the type of agar with its characteristic:
Signup and view all the answers
What color do Salmonella colonies appear on Hektoen Enteric Agar?
Signup and view all the answers
Beta hemolytic bacteria cause incomplete breakdown of red blood cells.
Signup and view all the answers
What is the function of bile salts in selective media?
Signup and view all the answers
What does a yellow broth indicate in carbohydrate fermentation tests?
Signup and view all the answers
The oxidase reaction tests for the presence of lecithinase.
Signup and view all the answers
What enzyme is tested for in the egg-yolk reaction?
Signup and view all the answers
The enzyme that allows bacteria to reduce nitrate is called __________.
Signup and view all the answers
Match the biochemical test with its purpose:
Signup and view all the answers
In the indole production test, what color indicates a positive result?
Signup and view all the answers
A Durham tube is not necessary in carbohydrate fermentation tests.
Signup and view all the answers
Which type of media is Eosin Methylene Blue Agar classified as?
Signup and view all the answers
What is the role of phenol red in carbohydrate fermentation tests?
Signup and view all the answers
The Pour Plate Method is primarily used for isolating single colonies from solid media.
Signup and view all the answers
What is the purpose of the Phenol red broth with Durham tube in biochemical tests?
Signup and view all the answers
Tryptophan breakdown can be assessed using _____ and Kovacs reagent.
Signup and view all the answers
Which test indicates the ability of bacteria to produce and maintain acid products from glucose fermentation?
Signup and view all the answers
Match the following biochemical tests with their corresponding media:
Signup and view all the answers
Blood Agar is classified as selective media.
Signup and view all the answers
The _____ test determines if ammonia is produced when media containing urea is used.
Signup and view all the answers
Study Notes
Culturing Bacteria
- A culture is a group of microorganisms that grow in a medium.
- A medium is a solid, liquid, or semi-solid that supports the growth of microorganisms.
- Common media include broth, agar, and enriched media.
- Microbiologists can identify microorganisms by studying their characteristics and culturing them in appropriate media.
Culturing Bacteria
- Pure cultures contain cells from a single species of an organism.
- The chosen medium must meet the organism's enzyme and nutritional requirements.
- Streak plating is used to isolate pure samples.
- Pour plating is used to count bacteria and obtain pure cultures.
- Serial dilution is a technique involved in the pour plate method.
- Serial dilution helps isolate single colonies for further study.
Estimating Bacterial Numbers
- Choose a plate with between 30 and 300 colonies for accurate counting.
- Multiply the number of colonies by the reciprocal of the dilution factor to estimate total bacteria.
- Express the result in scientific notation.
Culture Media
- Culture media contains all nutrients needed for bacterial growth.
- Microbiologists must understand the nutritional needs of bacteria to provide them with the necessary nutrients in an artificial environment.
- No single media supports the growth of all bacteria.
- Anaerobic conditions may be needed for certain species.
Common Human Pathogenic Enterobacteriaceae
- This table lists several common human pathogenic Enterobacteriaceae species and their associated genus.
Differential Media
- Differential media allow for the differentiation of bacteria based on their characteristics.
-
Eosin Methylene Blue (EMB) Agar:
- Detects fecal coliform bacteria.
- Contains Eosin Y and Methylene blue as pH indicators.
- Inhibits Gram-positive bacteria.
- Fermentable carbohydrates include lactose and sucrose.
- Rapid fermenters produce green metallic colonies.
- Medium fermenters produce pink colonies.
- Non-coliforms produce colorless colonies.
-
MacConkey Agar:
- Selective and differential.
- Selects for Gram-negative bacteria.
- Contains lactose, crystal violet, bile salts, and neutral red indicator.
- Lactose fermenters produce red colonies.
- Non-lactose fermenters produce colorless colonies.
-
Hektoen Enteric Agar:
- Selective and differential.
- Selects and differentiates Salmonella and Shigella from other Enterobacteriaceae.
- Contains bile salts and bromothymol blue, inhibiting Gram-positive bacteria.
- Ferrous compounds are present to indicate hydrogen sulfide gas production.
- Salmonella colonies produce transparent green colonies with black centers.
- Shigella colonies produce transparent green colonies.
- Lactose fermenters produce salmon-colored colonies.
-
Mannitol Salt Agar:
- Selective and differential.
- Isolates Staphylococci (Gram-positive bacteria).
- Contains a high salt concentration (7.5%), inhibiting halophobic growth.
- Pathogenic Staphylococci ferment mannitol, producing yellow colonies.
- Non-pathogenic Staphylococci do not ferment mannitol, producing pink colonies.
Selective Media
-
Bismuth Sulfite Agar:
- Isolates Salmonella typhi (typhoid fever).
- Contains bismuth sulfite and Brilliant green indicators, inhibiting Gram-positive and coliform bacteria.
- The presence of S. typhi reduces sulfide, resulting in black or green metallic colonies.
Enrichment Media
-
Tryptic Soy Agar and Broth (TSA/B):
- General purpose non-selective media.
- Provides ample nutrients for a wide range of bacterial species.
- Frequently used in general microbiology labs.
-
Blood Agar:
- Contains general nutrients and 5% bovine (cow or sheep) blood.
- Differentiates between hemolytic and non-hemolytic bacteria.
- Alpha hemolytic bacteria: partially break down RBCs and produce a green color (e.g., Streptococcus pneumoniae).
- Beta hemolytic bacteria: completely break down RBCs and cause clearing (e.g., Staphylococcus sp. and Streptococcus sp.).
- Gamma hemolytic bacteria: can grow but do not break down RBCs (e.g., Staphylococcus sp.).
-
Chocolate Agar:
- Blood is added to tryptic soy agar and heated, causing the blood to lyse and release the nutrients.
Biochemical Tests
-
Carbohydrate Fermentation Tests:
- Use phenol red broth with a Durham tube.
- Detects the fermentation of specific carbohydrates.
- Positive result: broth turns yellow (acidic).
- Negative result: broth stays red/pink (neutral).
- Gas production is indicated by bubbles in the Durham tube.
-
Litmus Test:
- Uses litmus milk medium with litmus indicator.
- Evaluates a bacteria's ability to use different milk products.
- Color changes, gas production, or curdling of the media indicate positive results.
-
Indole Production:
- Uses tryptophan broth and Kovacs reagent.
- Detects the ability of bacteria to break down tryptophan into indole.
- Positive result: pink color forms on the surface of the broth.
-
Methyl Red Test:
- Uses Methyl Red-Voges-Proskauer (MRVP) media and Methyl Red solution.
- Detects the ability of bacteria to produce and maintain acidic products from glucose fermentation.
- Positive result: red color remains after adding the Methyl Red solution.
-
Urease Test:
- Uses media containing urea.
- Detects the enzyme urease.
- Positive result: ammonia production turns the litmus indicator basic (color change).
-
Nitrate Reduction:
- Uses media containing nitrate.
- Detects the enzyme nitrate reductase.
- Positive result: the production of nitrites is observed.
-
Oxidase Reaction:
- Uses TSA/B and oxidase reagent.
- Detects the presence of cytochrome-oxidase, a key enzyme in the electron transport chain (ETC).
- Positive result: a blue color appears.
- This test helps determine a bacteria's ability to produce energy aerobically using the ETC.
-
Egg-Yolk Reaction:
- Uses egg-yolk agar.
- Detects the enzyme lecithinase, which destroys animal tissue.
- Positive result: a white precipitate forms around the colonies (indicates lecithinase activity).
Recall - Pour Plate Method
- A serial dilution is performed from a liquid specimen.
- Each dilution is used to pour a plate inoculated with bacteria in agar.
- Colonies are counted after incubation.
- A plate with single, pure colonies is selected for further growth.
Studying That Suits You
Use AI to generate personalized quizzes and flashcards to suit your learning preferences.
Related Documents
Description
This quiz covers the fundamentals of culturing bacteria, including the types of media used and methods of isolating pure cultures. You'll explore techniques such as streak plating and pour plating, along with estimating bacterial numbers through colony counts. Test your understanding of these essential microbiological practices.