1.1 Microbiology Fundamentals for Central Service

Key Takeaways

  • Microorganisms include bacteria, viruses, fungi, protozoa, and prions — each with different characteristics and resistance levels
  • Bacteria are classified as gram-positive or gram-negative based on their cell wall structure and staining characteristics
  • Bacterial spores (endospores) are the most resistant form of microbial life and require sterilization to destroy
  • Viruses require a host cell to replicate and range from easily killed (enveloped) to highly resistant (non-enveloped)
  • Fungi include yeasts and molds; they can cause opportunistic infections in immunocompromised patients
  • Prions are misfolded proteins that cause fatal neurological diseases (CJD) and cannot be destroyed by standard sterilization
  • Biofilm is a community of microorganisms that adhere to surfaces and produce a protective matrix, making them extremely difficult to remove
  • Understanding microbial resistance levels is essential for selecting the correct reprocessing method
Last updated: March 2026

Microbiology Fundamentals for Central Service

Understanding microbiology is the scientific foundation of everything Central Service technicians do. Every cleaning, disinfection, and sterilization process is designed to eliminate or reduce microorganisms to safe levels. Without understanding what you are fighting, you cannot fight it effectively.


Types of Microorganisms

1. Bacteria

Bacteria are single-celled organisms that can survive independently. They are classified by:

  • Shape: Cocci (round), bacilli (rod), spirilla (spiral)
  • Gram stain: Gram-positive (thick cell wall, stains purple) vs. gram-negative (thin cell wall with outer membrane, stains pink)
  • Oxygen requirement: Aerobic (needs oxygen), anaerobic (grows without oxygen), facultative (either)
  • Spore formation: Some bacteria (Bacillus, Clostridium) form endospores — dormant, highly resistant survival structures

Key bacteria in healthcare:

OrganismTypeSignificance
Staphylococcus aureus (MRSA)Gram-positive coccusLeading cause of surgical site infections
Clostridioides difficile (C. diff)Gram-positive, spore-formingHealthcare-associated diarrhea; spores resist many disinfectants
Pseudomonas aeruginosaGram-negative bacillusThrives in moist environments; biofilm former
Escherichia coliGram-negative bacillusIndicator organism for contamination
Mycobacterium tuberculosisAcid-fast bacillusResistant to many disinfectants; requires special protocols

2. Viruses

Viruses are obligate intracellular parasites — they cannot replicate without a host cell.

  • Enveloped viruses (HIV, Hepatitis B, influenza): Surrounded by a lipid envelope; easier to kill with disinfectants
  • Non-enveloped viruses (norovirus, adenovirus): No lipid envelope; more resistant to disinfection

Key viruses in healthcare:

VirusTypeConcern
Hepatitis B (HBV)Enveloped, DNABloodborne; survives on surfaces for 7+ days
Hepatitis C (HCV)Enveloped, RNABloodborne; major occupational exposure risk
HIVEnveloped, RNABloodborne; relatively fragile outside the body
NorovirusNon-enveloped, RNAEnvironmental contamination; difficult to kill

3. Fungi

Fungi include yeasts (single-celled) and molds (multicellular with hyphae).

  • Cause opportunistic infections in immunocompromised patients
  • Examples: Candida albicans (yeast), Aspergillus (mold)
  • Fungal spores are moderately resistant to disinfection

4. Protozoa

Protozoa are single-celled organisms that can form protective cysts.

  • Cryptosporidium cysts are resistant to standard chlorine disinfection
  • Less commonly encountered in CS but important in water quality discussions

5. Prions

Prions are misfolded proteins — not living organisms — that cause transmissible spongiform encephalopathies (TSEs):

  • Creutzfeldt-Jakob Disease (CJD) is the most relevant to CS
  • Prions are NOT destroyed by standard steam sterilization, EtO, hydrogen peroxide, or radiation
  • Special reprocessing protocols are required for suspected prion-contaminated instruments:
    • Immerse in 1N sodium hydroxide (NaOH) for 1 hour, then steam sterilize at 270°F (132°C) in a gravity displacement cycle for 1 hour
    • OR use a prevacuum sterilizer at 275°F (134°C) for 18 minutes after NaOH soak
    • Single-use instruments may be preferred for high-risk procedures

Biofilm — The Hidden Danger

Biofilm is a structured community of microorganisms enclosed in a self-produced extracellular polymeric substance (EPS) matrix that adheres to surfaces.

Why Biofilm Matters in CS:

  • Forms within minutes to hours after contamination
  • Is 1,000–1,500 times more resistant to antimicrobials than free-floating (planktonic) bacteria
  • Standard cleaning and sterilization may not penetrate mature biofilm
  • Commonly forms in lumens, channels, hinges, and crevices of surgical instruments

Preventing Biofilm Formation:

  1. Point-of-use treatment — keep instruments moist and wipe off gross soil immediately after use
  2. Prompt transport to decontamination (do not allow instruments to dry with soil)
  3. Thorough cleaning before disinfection or sterilization
  4. Enzymatic detergents break down organic matter that feeds biofilm

Remember: You cannot sterilize a dirty instrument. Cleaning is the most critical step in reprocessing because sterilization cannot penetrate biofilm or dried organic matter.


Order of Microbial Resistance (Most to Least Resistant)

Understanding resistance levels guides reprocessing decisions:

LevelOrganism TypeKilled By
Most resistantPrionsSpecial protocols (NaOH + extended steam)
Bacterial spores (Bacillus, Clostridium)Sterilization only
Mycobacteria (TB)High-level disinfection or sterilization
Non-enveloped viruses (norovirus)Intermediate to high-level disinfection
FungiIntermediate-level disinfection
Vegetative bacteria (Staph, Strep, E. coli)Low to intermediate-level disinfection
Least resistantEnveloped viruses (HIV, HBV)Low-level disinfection

This hierarchy is sometimes called the "hierarchy of microbial resistance" and is fundamental to understanding why different reprocessing methods exist.

Test Your Knowledge

Which type of microorganism is the MOST resistant to standard sterilization methods?

A
B
C
D
Test Your Knowledge

Biofilm on surgical instruments is dangerous because:

A
B
C
D
Test Your Knowledge

Which of the following bacteria is gram-positive, spore-forming, and a leading cause of healthcare-associated diarrhea?

A
B
C
D
Test Your Knowledge

The single most important step in preventing biofilm formation on surgical instruments is:

A
B
C
D