Podcast
Questions and Answers
Match the following bacteria with their fermentation type:
Match the following bacteria with their fermentation type:
Escherichia = Lactose Fermenter Shigella = Non-Lactose Fermenter Klebsiella = Lactose Fermenter Salmonella = Non-Lactose Fermenter
Match the following agar types with their characteristics:
Match the following agar types with their characteristics:
EMB Agar = Selective and differential XLD Agar = Detects hydrogen sulfide (H2S) Hektoen Enteric Agar = Targets Salmonella/Shigella SS Agar = Inhibits gram-positive organisms
Match the following non-lactose fermenters with their characteristics:
Match the following non-lactose fermenters with their characteristics:
Salmonella = Produces H2S and black center Shigella = Does not produce H2S Proteus = Non-Lactose Fermenter Yersinia = Non-Lactose Fermenter
Match the following lactose fermenters with their appearance on XLD agar:
Match the following lactose fermenters with their appearance on XLD agar:
Match the following fermentation results with the media type:
Match the following fermentation results with the media type:
Match the following carbohydrates found in XLD agar:
Match the following carbohydrates found in XLD agar:
Match the following colors with their meaning in Hektoen Enteric Agar:
Match the following colors with their meaning in Hektoen Enteric Agar:
Match the following Enterobacteriaceae species with their classification:
Match the following Enterobacteriaceae species with their classification:
Match the following late lactose fermenters with their initial characteristics:
Match the following late lactose fermenters with their initial characteristics:
Match the following characteristics with the correct type of bacteria:
Match the following characteristics with the correct type of bacteria:
Match the following infections with the corresponding Enterobacteriaceae species:
Match the following infections with the corresponding Enterobacteriaceae species:
Match the following media with their use in Enterobacteriaceae identification:
Match the following media with their use in Enterobacteriaceae identification:
Match the following tests with their characteristics in Enterobacteriaceae:
Match the following tests with their characteristics in Enterobacteriaceae:
Match the following terms with their descriptions:
Match the following terms with their descriptions:
Match the following features with the Enterobacteriaceae species:
Match the following features with the Enterobacteriaceae species:
Match the following terms with their key characteristics:
Match the following terms with their key characteristics:
Match the following bacterial tests with their primary purpose:
Match the following bacterial tests with their primary purpose:
Match the bacterial enzymes with their functions:
Match the bacterial enzymes with their functions:
Match the terms with their definitions:
Match the terms with their definitions:
Match the test conditions with their results:
Match the test conditions with their results:
Match the media usage with their testing purpose:
Match the media usage with their testing purpose:
Match the products of glucose metabolism pathways:
Match the products of glucose metabolism pathways:
Match the characteristics with their respective metabolic pathways:
Match the characteristics with their respective metabolic pathways:
Match the enzyme-related features with their corresponding information:
Match the enzyme-related features with their corresponding information:
Match the following compounds with their associated reactions or results:
Match the following compounds with their associated reactions or results:
Match the following amino acids with their corresponding decarboxylase enzymes:
Match the following amino acids with their corresponding decarboxylase enzymes:
Match the following tests with their specific purposes:
Match the following tests with their specific purposes:
Match the following results with their corresponding color changes:
Match the following results with their corresponding color changes:
Match the following bacteria genera with their ability to produce specific enzymes:
Match the following bacteria genera with their ability to produce specific enzymes:
Match the following components of Moeller’s broth with their functions:
Match the following components of Moeller’s broth with their functions:
Match the following conditions necessary for decarboxylation with their descriptions:
Match the following conditions necessary for decarboxylation with their descriptions:
Match the following products with their respective sources in the deamination test:
Match the following products with their respective sources in the deamination test:
Match the following tests with what they detect or indicate:
Match the following tests with what they detect or indicate:
Match the carbohydrate fermentation results with their corresponding descriptions:
Match the carbohydrate fermentation results with their corresponding descriptions:
Match the metabolic products with the tests where they are detected:
Match the metabolic products with the tests where they are detected:
Match the characteristics of indole-positive bacteria:
Match the characteristics of indole-positive bacteria:
Match the colony colors on various agar media with the organisms they indicate:
Match the colony colors on various agar media with the organisms they indicate:
Match the following tests with their results:
Match the following tests with their results:
Match the following tests with their purposes:
Match the following tests with their purposes:
Match the following positive results with their corresponding tests:
Match the following positive results with their corresponding tests:
Match the following negative results with their respective tests:
Match the following negative results with their respective tests:
Match the following tests with the correct reagents used:
Match the following tests with the correct reagents used:
Match the following microorganisms with their positive tests:
Match the following microorganisms with their positive tests:
Match the following limitations with their respective tests:
Match the following limitations with their respective tests:
Match the following outcomes with their definitions:
Match the following outcomes with their definitions:
Flashcards
Enterobacteriaceae Family
Enterobacteriaceae Family
A family of gram-negative bacteria found in many environments including the human gut; many members are opportunistic pathogens.
Opportunistic Pathogens
Opportunistic Pathogens
Bacteria that become pathogenic in situations where the host's immune system is weakened or the bacteria are in unusual locations.
Escherichia coli
Escherichia coli
A common opportunistic pathogen; often found in the gut and can cause UTIs, septicemia and other infections.
Klebsiella pneumoniae
Klebsiella pneumoniae
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Proteus mirabilis
Proteus mirabilis
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MacConkey Agar (MAC)
MacConkey Agar (MAC)
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Glucose Fermentation
Glucose Fermentation
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Nitrate Reduction
Nitrate Reduction
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Oxidase Negative
Oxidase Negative
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Motility (Enterobacteriaceae)
Motility (Enterobacteriaceae)
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Lactose Utilization
Lactose Utilization
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Lactose
Lactose
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β-galactoside permease
β-galactoside permease
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β-galactosidase
β-galactosidase
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ONPG
ONPG
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ONPG Test
ONPG Test
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Glucose Metabolism Pathways
Glucose Metabolism Pathways
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Oxidation-Fermentation (O/F) Test
Oxidation-Fermentation (O/F) Test
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Lactose Fermenters
Lactose Fermenters
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Non-Lactose Fermenters
Non-Lactose Fermenters
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Late Lactose Fermenters (LLF)
Late Lactose Fermenters (LLF)
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EMB Agar
EMB Agar
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Hektoen Enteric (HE) Agar
Hektoen Enteric (HE) Agar
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Salmonella
Salmonella
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Shigella
Shigella
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Xylose Lysine Deoxycholate (XLD) Agar
Xylose Lysine Deoxycholate (XLD) Agar
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Lysine Decarboxylase
Lysine Decarboxylase
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Salmonella colony on XLD
Salmonella colony on XLD
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Shigella colonies on XLD
Shigella colonies on XLD
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Salmonella/Shigella Agar (SS Agar)
Salmonella/Shigella Agar (SS Agar)
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Decarboxylase Test
Decarboxylase Test
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Decarboxylase Principle
Decarboxylase Principle
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Dihydrolase Test
Dihydrolase Test
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Positive Decarboxylase Result
Positive Decarboxylase Result
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Negative Decarboxylase Result
Negative Decarboxylase Result
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Diacetyl Test
Diacetyl Test
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Phenylalanine Deaminase Test
Phenylalanine Deaminase Test
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Moeller's Broth
Moeller's Broth
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Mixed acid fermentation
Mixed acid fermentation
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Voges-Proskauer test
Voges-Proskauer test
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Glucose & lactose fermentation
Glucose & lactose fermentation
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Indole production
Indole production
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Tryptophanase
Tryptophanase
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API system
API system
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XLD agar black colonies
XLD agar black colonies
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HEK agar orange colonies
HEK agar orange colonies
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MAC agar lactose fermenters
MAC agar lactose fermenters
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SS agar pink colonies
SS agar pink colonies
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Ferric Chloride Test Result (Positive)
Ferric Chloride Test Result (Positive)
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Ferric Chloride Test Result (Negative)
Ferric Chloride Test Result (Negative)
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Tryptophan Deaminase Test (Positive)
Tryptophan Deaminase Test (Positive)
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Tryptophan Deaminase Test (Negative)
Tryptophan Deaminase Test (Negative)
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Citrate Utilization Test (Positive)
Citrate Utilization Test (Positive)
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Citrate Utilization Test (Negative)
Citrate Utilization Test (Negative)
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Indole Test (Positive, Ehrlich's Reagent)
Indole Test (Positive, Ehrlich's Reagent)
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Indole Test (Positive, Kovac's Reagent)
Indole Test (Positive, Kovac's Reagent)
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Indole Test (Positive, Spot Test)
Indole Test (Positive, Spot Test)
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Motility Test (Positive)
Motility Test (Positive)
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Ornithine Decarboxylase (Positive)
Ornithine Decarboxylase (Positive)
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Study Notes
Overview of Enterobacteriaceae Identification Methods
- Lecture 9 covers identifying Enterobacteriaceae
- Includes information from Bailey & Scott's (Chapter 12, 19) and Diagnostic Microbiology (Chapter 9, 19)
- Lecture introduces Enterobacteriaceae, common identification methods, special media and test systems.
Enterobacteriaceae
- A large family of bacteria, also known as enterics.
- Common clinical isolates in acute care primarily include Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis.
Bacterial Species and Infections
- Table 19.3 lists various bacterial species with the diseases they commonly cause.
- Examples include Escherichia coli linked to bacteriuria, septicemia, and neonatal sepsis.
- Some are opportunistic pathogens that cause infections in non-normal areas, such as the urinary tract or wounds.
Clinical Significance
- Two main categories of clinical significance:
- Opportunistic pathogens (covered in this lecture): These bacteria are part of normal flora but can cause infections in unusual sites.
- Primary pathogens: These bacteria always cause disease. Examples include Salmonella, Shigella, and Yersinia.
General Characteristics of Enterobacteriaceae
- All ferment glucose.
- All reduce nitrates to nitrites.
- All are oxidase-negative.
- Most are motile at body temperature, except for Klebsiella, Shigella, and Yersinia.
- Lack remarkable colony morphologies on typical lab media.
Enterobacteriaceae - Identification
- Cellular morphology (gram-negative rods/coccobacilli) is not very helpful for identification except to rule out other organisms.
- Growth conditions are facultatively anaerobic.
- Molecular methods are modern methods for identification.
- Macroscopic morphology on BAP is not very helpful for identification, except to rule out other organisms.
- Larger, moist, gray colonies are sometimes mucoid; selective plates are used for identifying pathogenic species in stool samples.
MacConkey Agar
- A selective and differential medium for Enterobacteriaceae.
- Inhibits gram-positive bacteria; lactose fermentation is the key differentiator.
- Colonies of lactose fermenters appear pink/red (LF); non-lactose fermenters are yellow (NLF). Some may exhibit delayed lactose fermentation (LLF), initially appearing yellow but turning pink/red after 48 hours.
EMB Agar
- A selective and differential medium often used for Enterobacteriaceae identification.
- Methylene blue inhibits gram-positive bacteria; lactose and sucrose fermentation differentiate.
Hektoen Enteric Agar (HE)
- Selective and differential medium specific for Salmonella and Shigella.
- Bile salts and other inhibitory agents reduce gram-positive growth; lactose and sucrose fermentation aid differentiation.
- Orange colonies often indicate non-pathogenic organisms.
- Black colonies with a central black discoloration indicate Salmonella.
- Blue-green colonies indicate possible target pathogens, like Shigella.
Xylose Lysine Deoxycholate Agar (XLD)
- Another medium targeted specifically at Salmonella and Shigella identification.
- Selective for gram-negative bacteria; less than MacConkey or HE.
- Lactose, sucrose, and xylose fermentation; lysine decarboxylation; and H2S production differentiate.
- Yellow colonies (non-pathogens)
- Red colonies with black centers (target pathogens).
- Colorless/red colonies (Salmonella/Shigella)
Salmonella Shigella Agar (SS)
- A medium for isolating and differentiating Salmonella and Shigella.
- Selective for gram-negative bacteria; lactose and neutral red differential.
- Lactose fermenters (pink); non-lactose fermenters (colorless)
- This allows identification of non-pathogenic bacteria from possible pathogens.
CIN Media
- A medium for isolates that have Yersinia spp. or non-fermenting organisms.
- Inhibitory agents (cefsulodin, irgasan, novobiocin, bile salts, and crystal violet) reduce growth. Mannitol with phenol red are the differential differentiators.
O/F Basal Media
- Useful for classifying bacteria as oxidizers or fermenters.
- Two tubes are used:
- Aerobic tube
- Anaerobic tube (covered by mineral oil).
Triple Sugar Iron Agar (TSI)
- Detects production of acid from fermentation of glucose, lactose, and sucrose, and also hydrogen sulfide.
- The butt of the tube and the slant are both used for interpretation.
- Gas may also be noted.
- Examples of TSI reactions are noted.
- Several organisms have their own specific reactions and are highlighted.
Glucose Metabolism
- The Embden-Meyerhof pathway is the essential pathway.
- Other pathways are mixed acid or butylene glycol pathways, which have their own specific tests.
Methyl Red(MR) test
- This test determines if mixed acids are produced during glucose metabolism.
- Positive results are indicated by a red color.
Voges-Proskauer (VP) test
- Identifies organisms that utilize the butylene glycol pathway.
- Positive is indicated by the formation of a red complex.
Decarboxylase and Dihydrolase Tests
- Used to distinguish decarboxylase-producing Enterobacterales from other Gram-negative rods; specific for amino acids (e.g., lysine and ornithine).
- Moeller's broth is an example of a medium used in these tests, using appropriate pH indicator.
- A positive result is marked by a color change; a negative result does not change or is a slight change in color.
Phenylalanine Deaminase Test
- Used to test if bacterial species can deaminate phenylalanine.
- Positive is indicated by a green color change.
Tryptophan Deaminase Test
- Identifies organisms that can deaminate tryptophan and produce indole
- Positive is indicated by a reddish color.
Citrate Utilization Test
- Tests if an organism can utilize sodium citrate as a carbon source.
- Positive is indicated by a blue color change.
Indole test
- A test to detect if indole production occurs from tryptophan.
- Positive is indicated by a change from colorless to a reddish brown color.
Motility-Indole-Ornithine (MIO) Agar
- A semi-solid medium useful for differentiating species of Klebsiella, Enterobacter, and Serratia. This is done by testing for motility, ornithine decarboxylase, and indole formation.
Nitrate and Nitrite Reduction
- This reaction detects the ability of an organism to reduce nitrate.
- A positive result is marked when a color is not seen or if gas is seen after adding zinc.
- Negative result turns a reddish brown color.
Urease (Christensen's) Test
- Determines if an organism can produce urease.
- Positive result shows a bright pink color.
Multi-test systems
- Provide comprehensive identification.
- Can be pure culture or don't require one.
- pH, enzymatic activity, carbon utilization, visual detection of characters, or molecular methods.
- API 20E system is an example.
Rapid identification systems
- Offer quicker results.
- Several types of rapid methods exist, using different techniques, and include immunochemical methods, mass spectrometry, and molecular biological methods.
Knowledge Checks
- Answers to relevant knowledge checks.
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