Bacterial culture methods

Bacterial culture methods

Bacterial culture methods

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Culture Fundamentals - The Bacterial Buffet

  • Culture Medium: A nutrient preparation (broth or solid agar) used to grow microorganisms.
  • Minimal Medium: Contains basic requirements for non-fastidious microbes to grow; simple salts and a carbon source.
  • Enriched Medium: Fortified with specific growth factors like blood, serum, or vitamins to support fastidious organisms (e.g., Chocolate agar for Haemophilus).
  • Selective Medium: Suppresses the growth of unwanted microbes while encouraging the growth of desired ones (e.g., Thayer-Martin agar for Neisseria).
  • Differential Medium: Distinguishes microorganisms based on specific biochemical characteristics, often via a color indicator (e.g., MacConkey agar for lactose fermentation).

⭐ Many media are both selective and differential. MacConkey agar, for instance, inhibits Gram-positive bacteria (selective) while differentiating lactose-fermenting bacteria, which turn pink (differential).

Selective & Differential Media - Picky Eater Plates

  • Selective Media: Favors the growth of specific microbes while inhibiting others.
  • Differential Media: Distinguishes between different groups of bacteria based on their biochemical characteristics.

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MediaSelective ForDifferential ForKey Reactions
MacConkey AgarGram (-)Lactose FermentationFermenters → Pink colonies
Eosin-Methylene Blue (EMB) AgarGram (-)Lactose FermentationE. coli → Green metallic sheen
Hektoen Enteric (HE) AgarGram (-)Lactose & H₂SLactose fermenters → Yellow/Pink
H₂S producers → Black precipitate
Mannitol Salt Agar (MSA)Staphylococcus (high 7.5% NaCl)Mannitol FermentationS. aureus → Yellow colonies

📌 Mnemonic: MacConkey = Crystal violet inhibits Gram (+). EMB = Excellent Metallic Blue for E. coli.

Special Culture Techniques - The Air Snobs

  • Bacteria are categorized by their oxygen requirements, impacting culture methods.
  • Obligate Aerobes: Require $O_2$ for metabolism. 📌 Nagging Pests Must Breathe (Nocardia, Pseudomonas, Mycobacterium, Bacillus).
  • Obligate Anaerobes: $O_2$ is toxic. Lack catalase and/or superoxide dismutase. 📌 Anaerobes Can't Breathe Air (Actinomyces, Clostridium, Bacteroides).
  • Facultative Anaerobes: Use $O_2$ but can grow without it. Most bacteria.

Anaerobic Jar with GasPak System Diagram

Clostridium perfringens grown on blood agar demonstrates a characteristic double zone of hemolysis (target hemolysis), a key identification feature resulting from alpha and theta toxins.

High‑Yield Points - ⚡ Biggest Takeaways

  • Selective media inhibit unwanted bacteria, while differential media distinguish them by metabolic activity.
  • MacConkey agar is both: it selects for Gram-negatives and differentiates lactose fermenters (pink colonies).
  • Blood agar differentiates bacteria based on hemolysis: α-hemolysis (partial, green), β-hemolysis (complete, clear), and γ-hemolysis (none).
  • Chocolate agar, with its lysed RBCs, supports fastidious organisms like H. influenzae and Neisseria.
  • Thayer-Martin agar is a selective chocolate agar with antibiotics, specifically for isolating Neisseria species.

Practice Questions: Bacterial culture methods

Test your understanding with these related questions

A 42-year-old woman comes to her primary care physician with 2 days of fever and malaise. She also says that she has a painful red lesion on her left hand that she noticed after shucking oysters at a recent family reunion. Physical exam reveals a well-demarcated swollen, tender, warm, red lesion on her left hand. Pressing the lesion causes a small amount of purulent drainage. The material is cultured and the causative organism is identified. Which of the following characteristics describes the organism that is most associated with this patient's mechanism of infection?

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Flashcards: Bacterial culture methods

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Indicator (differential) media can also yield color changes due to differences in _____ of certain organisms

TAP TO REVEAL ANSWER

Indicator (differential) media can also yield color changes due to differences in _____ of certain organisms

metabolism

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