CULTURE SYSTEM

• CULTURE MEDIA CONSISTENCY

  1. LIQUID - ex. Broth
  2. SEMI-SOLID - contain 0.5-1% agar; used for motility testing
  3. SOLID
• BACTERIAL CULTURE TISSUE COMPOSITION

  Mccoy cell: from mouse cell line; ex. Chlamydia

  Vero cell: African green monkey

  A549 cell: lung carcinoma

  Hela cell: cervical carcinoma

  Hep-2 cell: Laryngeal carcinoma
• Bacterial Culture for motility test

  Semi-solid medium
• Virally cultures

  Tissue
• SIMPLE/GENERAL PURPOSE


  support growth of non-fastidious organism

  ex: Nutrient Agar or Broth, Trypticase soy or Broth
• SELECTIVE MEDIUM

  SELECT growth of PARTICLE ORGANISM and PREVENT OTHER ORGANISM thru INHIBTORS

  INHIBITORS:

  Lowenstein Jensen - M. tuberculosis

  Phenylethyl Alcohol - Gram Positive organims

  Thayer Martin, Martin Lewis, NYC - N. gonorrhea
• ENRICHED MEDIUM


  culture media contains NUTRIENT SUPPLEMENTS

  ex: BAP - 5% defibrinated Sheep Blood (Horse/Rabbit)

  CAP - lysed blood, used for Neisseria spp and Haemophilus spp
• ENRICHMENT MEDIUM 


  enhance growth of microorganism

  ex: ALKALINE PEPTONE WATER - used for VIBRIO

  SELENITE AND TETRATHIONATE BROTH - used for Salmonella and Shigella
• SELECTIVE - DIFFERENTIAL MEDIUM


  allowing certain types of organism to grow and differentiate and inhibit the growth of other organisms

  Ex: MANNITO SALT AGAR - S. aures, S. epidermidis, S. saprophyticus

  THIOSULFATE CITRATE BILE SALTS - Vibrio spp.

  SALMONELLA-SHIGELLA AGAR
• DIFFERENTIAL MEDIUM 


  provides DISTINCT COLONIAL APPEARANCE of microorganism to know their IDENTIFICATION

  Ex: TSI - 3 SUGARS: Lactose, Glucose, Sucrose
• HEMOLYSIS ON SHEEP BLOOD AGAR

  ALPHA HEMOLYSIS - partial hemolysis; Green-Zone around colony

  BETA HEMOLYSIS - complete hemolysis

  GAMMA HEMOLYSIS - no hemolysis
• ALPHA HEMOLYSIS ORGANISM

  "SEV"

  S. pnuemoniae

  Viridans streptococci

  Some Enterococci
• BETA HEMOLYSIS ORGANIMS


  Group A Strep - S. pyogenes

  Group B Strep - S. agalactiae

  L. monocytogenes
• GAMMA HEMOLYSIS ORGANISMS

  Enterococci
• TARGET HEMOLYSIS

  aka "Double Zone Hemolysis"

  • Clostridium perfringens - cause GAS GANGRENE DISEASE or MYONECROSIS
• ALPHA PRIME (a') 


  aka "WIDE ZONE HEMOLYSIS" small area of intact rbc around colony surrounded by wider zone of complete hemolysis

  • VIRIDANS GROUP
• SHEEP BLOOD AGAR MEDIUM


  enriched medium for growth of most NON-FASTIDIOUS ORGANISM

  • TRYPTIC SOY AGAR with 5% SHEEP BLOOD

  DIFFERENTIATION OF HEMOLYSIS
• CHOCOLATE AGAR MEDIUM


  enriched media for Haemophilus spp and Neisseria spp

  • supplies X and V factor with INCREASED CO2 INCUBATION

  X factor - "HEMIN"

  V factor - "NAD"
• COLUMBIA COLISTIN NALIXIDIC ACID AGAR (CNA)

  Selective medium for GRAM POSITIVE MICROORGANISMS

  • Colistin and Nalixidic - suppressed most Gram Negative organism
  • contain 5% Sheep Blood
• PHENYLETHYL ALCOHOL AGAR (PEA)


  selective medium for Gram Positive cocci and Anaerobic Gram Negative Rods

  • PHENYLETHYL ALCOHOL inhibits enteric Gram Negative Rods
  • Contain 5% sheep blood
• STREPTOCOCCAL SELETIVE AGAR (SSA)


  Selective medium for BETA HEMOLYSIS ORGAMISMS: Group A strep. and Group B strep.
• EOSIN METHYLENE BLUE (EMB)

  Selective Differential medium for isolation of ENTERIC GRAM NEGATIVE RODS

  • ENTEROBACTERIACEAE

  INHIBITORS: EOSIN and METHYLENE BLUE - inhibits gram positive

  LACTOSE FERMENTERS - produces GREEN- BLACK or PURPLE

  NON- LACTOSE FERMENTS - COLORLESS
• Mac Conkey Agar

  Selective, differential medium for isolation of Enteric Gram- Negative Rods

  • ENTEROBACTERIACEAE
  • Contains: BILES SALTS and CRYSTAL VIOLET - inhibits gram positive

  LACTOSE FERMENTES: PINK

  NON-LACTOSE FERMENTS: colorless
• Sorbitol Mac Conkey Agar (SMAC)

  Selective medium for E. coli O157:H7 with colorless colonies

  • SORBITOL replace LACTOSE in MacConkey Agar

  Sorbitol doesnt ferment E. coli 0157:H7
• E. coli O157:H7 associated disease

  HUS (HEMOLYTIC UREMIC SYNDROME)
• HEKTOEN ENTERIC AGAR (HE)

  Selective Differential medium for isolation of ENTERIC PATHOGENS from stool

  INHIBITORS: Bile Salts, Bromthymol Blue, Acid fuchsin - inhibits normal GI FLORA

  • NON-PATHOGEN: orange-salmon pink
  • NON-LACTOSE FERMENTERS: green to blue-green
  • H2S +: BLACK PRECIPITATE

  NLF: Salmonella, Shigella - green color
• XYLOSE LYSINE DEOXYCHOLATE (XLD)

  Selective differential medium for isolation of SALMONELLA and SHIGELLA in stool.

  INHIBITOR: Deoxycholate - inhibits many gram negative rods and gram positive

  XLD 4 TYPES OF COLONIES:

  1. E. Coli - yellow
  2. Proteus - yellow with black center (H2S)
  3. Shigella - red/colorless
  4. Salmonella - red with black center (H2S)
• SALMONELLA-SHIGELLA AGAR

  Selective medium for Salmonella and Shigella

  INHIBITORS: Brilliant green, Bile Salts - inhibits COLIFORMS

  NON-LACTOSE FERMENTERS: Shigella, Salmonella

  • Salmonella - H2S (+)
  • Shigrlla - H2S (-)
• ISOLATION TEST for SALMONELLA and SHIGELLA

  1. SELENITE BROTH
  2. TETRATHIONATE BROTH
• CAMPYLOBACTER BLOOD AGAR 


  selective enrichment medium for isolation of campylobacter from stool

  • INCREASED CO2 @ 42 DEG C
• CAMPHYLOBACTER

  Microaerophilic - requires 5% Oxygen
• SELECTIVE MEDIA FOR N. gonorrhea and N. Meningitis

  1. THAYER MARTIN
  2. MODIFIED THAYER MARTIN
  3. MARTIN-LEWIS
  4. NYC
  5. GC-LEG
  6. JEMBEC PLATES
• MODIFIED THAYER MARTIN


  selective enrichement medium for recovery of N. gonorrhea and N. meningitidis from specimens with normal flora.

  ANTIBIOTICS: "VCN+T" - inhibit growth of other bacteria and fungi

  Vancomycin - Gram Positive

  Colistin - Gram Negative

  Nystatin - anti-fungal

  Trimethorim - inhibits swarming for Proteus
• MARTIN LEWIS


  Selective enrichment medium for recovery of N. gonorrhea and N. meningitidis from specimen with normal flora

  ANTIBIOTICS: "VCAT

  Vancomycin

  Colistin

  Anisomycin - antifungal; inhibit yeast better

  Trimethoprim
• NEW YORK CITY

  ANTIBIOTICS: "VCAbT"

  Vancomycin

  Colistin

  Amphotericin B

  Trimothoprim
• JEMBEC PLATES

  for transportaion and growth of N. gonorrhea

  • Polyethylene Bag and CO2 generating tablet