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26 Cards in this Set
- Front
- Back
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Process of day 1 for ID of Staph. |
Isolation of nasal with sterile swab and place in M-Staph broth. |
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Explain the process of the completed test. |
Inoculate half the colony (pink /violet of the ECC, into a SSLB and the other half of the colony on TSA slant. Incubate, confirm gas present , gram stain the slant for gram negative rod. |
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What is the purpose of examination of water? |
To test that the water is safe and potable. |
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What bacteria is used as an indicator ? |
E.coli as it is not found in soil or water, but in the intestines of humans and warm blooded animals. |
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What is water testing looking for? |
The presence of coliforms. |
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What are the characteristics of coliform? |
Gram negative, Rods, non-spore forming, and facultative anaerobes. They ferment lactose, to produce gas and acid. |
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What are the 3 test that are used to determine the coliform count in water. |
The Presumptive , confirmed, and completed test. |
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Explain the propose of the presumptive test. |
To see if gas is produced when Lactose broth is inoculated with water sample . If so, coliform May be present . The MPN determines the number of coliform per 100ml of water.
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Does a positive result from the presumptive test indicate presence of coliforms? |
No, the organism might be lactose fermenters. |
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Explain the purpose of the confirmed test |
It uses differential and selective media to confirm that the organisms from the positive lactose broths are gram negative and lactose fermenters. |
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What is the purpose of the completed test? |
This confirms that a colony from the confirmed test is a gram negative gas producer . |
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Explain the process of the presumptive test. |
5 double strength lactose broths (DSLB) -10 ml water 10 single strength lactose broths (SSLB) -5, 1ml of water , 5 -0.1ml of water Incubate -24 hours at 38 degrees Celsius Determine the MPN and positive gas production in sets of 3.
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Explain the process of the confirmed test |
This is performed after the calculation of MPN. Inoculate 3 different plates with one positive tube. Streak for isolation of colonies to determine organisms. |
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Eosin methylene blue (EMB) agar |
Isolates coliforms, the dyes eosin and methylene blue inhibit gram positive bacteria. The lactose encourages coliform growth through lactose fermentation which produces an acidic environment Colonies = metallic green colony |
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MacConkey agar |
Streak for isolation. Media contains bile salts and crystal violet which Inhibit gram positive bacteria. Media is lactose and pH Indictator neutral red-gives the lactose fermenters their pink to red color.precipitation of bile salts create the haziness. Colonies=pink/red hazy colonies |
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HardyCHROM ECC agar |
Chromogenic media that differentiates E. Coli from other coliforms. Added agents inhibit the growth of gram positive and non coliforms. Chromogenic substrates allow certain organisms to be distinguished by the color of colonies. Colonies = pink/violet |
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Purpose of Isolation and identification for staph |
Identify the unknown Staph from Nasal and unknown broth |
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Process of Day 2 of ID Staph with SM110. |
2 SM110, contains mannitol and sodium chloride . Streak for isolation of nasal and unknown S.aureus appear yellow or orange, and other colonies appear colorless. |
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Process of Day 5 of ID Staph with Coagulase |
This is done if mannitol is positive and beta hemolytic If Coagulase is still liquid = negative If Coagulase is still solid, semi-solid = positive |
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Bacterial transformation:pGLO |
To express amp resistance and GF protein.Genetic transformation means change caused by genes, and involves the insertion of a gene into an organism- to change the organisms trait. Genetic transformation used in agriculture, digest oils spills., gene therapy |
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Explain the process of bacterial transformation. |
Transform bacteria with a gene that codes for Green Fluorescent Protein (GFP). The bacteria will express their jellyfish gene and produce the fluorescent protein - causes them to glow a green color under ultraviolet light . |
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Explain the purpose of a plasmid . |
Bacteria can transfer plasmids back and forth - allowing them to share beneficial genes. Allows bacteria to adapt to new environments. Causes bacterial resistance to antibiotics .
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Bacterial conjugation purpose |
DNA from one cell is transferred to another cell to produce a recombinant cell. Plasmids can replicate independently , move from cell to cell. F (fertility) factor, regulates the formation of pili that can function as a bridge between two bacteria cells. Allowing the transfer of DNA from donor to recipient — conjugation |
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Explain the process of bacterial conjugation. |
E.coli strain I (resistant to streptomycin) strain II (resistant to ampicillin) Label 4 plates by Strain I, II, conjugation |
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What are the observations for pGLO, LB/amp/ara? |
It grew and it glows. Arabinose is the inducer which makes it glow and induces the operon. It has a plasmid = glow |
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When is the GFP on? |
Only when the operon is on |
