Title | 49153440 Bacterial Culture Media in Plate Tube |
---|---|
Author | Khae Rapinan |
Course | Medical Technology |
Institution | Our Lady of Fatima University |
Pages | 10 |
File Size | 900.8 KB |
File Type | |
Total Downloads | 97 |
Total Views | 620 |
BACTERIOLOGY 231PLATE MEDIAMEDIA CARBOHYDRATE INHIBITOR pH INDICATOR RESULT BAP Blood agarOutdated human blood or 5% defribinated sheep blood (preferred)Trypticase Soy Agar or Nutrient agar{ Nonselective agar for Hemolysis Differentiation }PINHEAD – Staphylococcus PINPOINT - StreptococcusAlpha hemol...
BACTERIOLOGY 231 PLATE MEDIA MEDIA
CARBOHYDRATE
INHIBITOR
pH INDICATOR
BAP Blood agar
Alpha hemolysis
RESULT Incomplete or partial lysis with greening S. pneumoniae
Outdated human blood or 5% defribinated sheep blood (preferred) Trypticase Soy Agar or Nutrient agar
Beta hemolysis { Nonselective agar for Hemolysis Differentiation }
S. pyogenes S. agalactiae
PINHEAD – Staphylococcus PINPOINT - Streptococcus
Gamma hemolysis
CAP Chocolate agar
Complete lysis with colorless zone [diffusion of hemoglobin into the surrounding medium]
Nonhemolytic
Hemolytic Lysed red blood cells Trypticase Soy Agar or Nutrient agar X factor (hemin) V factor or NAD (nicotinamide adenine dinucletide, coenzyme)
{Nonselective agar for Hemolysis Differentiation} PINHEAD – Staphylococcus PINPOINT – Streptococcus **S. aureus - 1° src. of X factor
Nonhemolytic
PEA Phenylethyl alcohol agar
Bacillus anthracis (24° incubation) **Most Bacillus sp. grow
{Selective & Differential agar} Gram (-) inhibitor Inhibits swarming of Proteus
MSA Mannitol salt agar
✔ Yellow colonies [pH 6.8]
Coagulase (+) S. aureus Some Micrococcus & Enterococcus
{Selective & Differential agar} Inhibits Gram (-)
Mannitol
7.5% NaCl
Phenol red
✖Small red colonies with red/purple zone [pH 8.4]
Coagulase (-) Staph. sp. (episodemidis/saprophiticus) Most Gram (-)
✔Pink to purple colonies with Green metallic sheen
E. coli (Rapid Lactose Fermenter - RLF)
Beef extract
EMB Eosin methylene blue agar {Selective & Differential agar} Inhibits Gram (+)
Lactose
Eosin Methylene blue
✔Pink to purple colonies with Green metallic sheen w/ or w/o black center (fisheye) ✔ Pink to purple colonies, mucoid
Enterobacter aerogenes (RLF) Motile*
Klebsiella pneumonia (RLF) Nonmotile*
✖Yellow to colorless (pink) colonies (NLF)
MAC MacConkey agar
✔Pink to red colonies [pH gray to black colonies Corynebacterium diptheriae
Francisella tularensis Legionella pneumophilia Clostridium difficile Prevotella melaninogenica 5-6 wks: LEPTOSPIRA 3-4 wks: BRUCELLA 2-3 wks: MYCOBACTERIUM BACTERIOLOGY 231
MODIFIED TYNSDALE AGAR >black colonies with brown halos LOEFFLER SERUM SLANT >enhance METACHROMATIC granules BLOOD CYSTEIN GLUCOSE AGAR BUFFERED CHARCOAL YEASE EXTRACT (BYCE) With L-cysteine CYCLOSERINE CEFOXITIN FRUCTOSE AGAR (CCFA) (Brucella) LAKED KANAMYCIN VANCOMYCIN 5% SHEEP BLOOD AGAR (LKV) BIPHASIC (2 phased media) >SEPTI-CHECK & OLD CASTAÑEDA *ideal for airborne microbes
TUBE MEDIA MEDIA
TSI Triple Sugar Iron agar
FORMATION
Butt Slant
CARBOHYDRATE
pH INDICATOR
Glucose [Butt] Lactose [Slant] Sucrose [Slant]
Phenol red
Decarboxylation (slant) = Alkaline (red)
H2S INDICATOR
RESULT A/AG
Ferrous ammonium sulfate
A/AG+ or K/AG+
A/AG or K/AG
K/AG+
A/A or K/A K/A
K/A+ K/K
Legend: A(yellow); K(red); G(gas production); +(H2S production)
E. coli Enterobacter aerogenes Pantoea agglomerans Klebsiella pneumonia Serratia Salmonella arizonae Citobacter freundii Salmonella arizonae Hafnia alvei Morganella morganii Morganella freundii Citrobacter koseri Serratia marcescens Hafnia alvei Proteus vulgaris Proteus mirabilis Salmonella paratyphi Salmonella choleraesuis Edwardsiella tarda Yersinia entericolitica Shigella sonnei Shigella dysenteriae Shigella flexneri Shigella boydii Salmonella typhi Pseudonmonas aeroginosa Alcaligenes
MEDIA LIA Lysine Iron Agar
FORMATION
Butt Slant
AMINO ACID
Lysine
CARBOHYDRATE
Glucose
pH INDICATOR
H2S INDICATOR
Bromcresol purple
Ferric ammonium citrate
RESULT
K/K
K/A
R/A *deaminase(+)
K/AG+
Legend: K(purple); A(yellow); R(red); +(H2S production)
E. coli Enterobacter aerogenes Klebsiella pneumonia Serratia marcescens Hafnia alvei Pantoea agglomerans Citobacter freundii Shigella dysenteriae Shigella flexneri Shigella boydii Proteus Providencia Morganella Salmonella typhimurium Salmonella arizonae Salmonella typhi Salmonella parayphi Salmonella choleraesuis Edwardsiella tarda
MEDIA
Indol Production Test
Methyl Red Test MIXED ACID PATHWAY
Voges Proskauer Test BUTYLENE GLYCOL PATHWAY
PRINCIPLE The organism oxidized tryptophan by trytophanase ↓ tryptophan ↓ pyruvic acid ↓ amino acid ↓ indol
The organism ferments glucose to pyruvic acid resulting in mixed acid products leading to decreased pH (4.55.5) of the medium and takes the color of the indicator
The organism ferments glucose producing acetylmethylcarbinol (acetoin) and converted to dimethyl/diacetyl in the presence of oxygen and KOH. KOH → reacts w/ GUANIDINE = color
FORMATION
MEDIUM
Butt
Tryptone broth
Butt
MRVP (Methyl Red Voges Methyl red indicator Proskauer) medium
Butt
MRVP
REAGENT ①Ehrlich’s Add: EtOH and/or XYLENE (xtract indol) then p-dab ②Kovac’s p-aminodimethyl benzaldehyde (p-dab) ③Spot Indole: rapid detection – filter paper:1% pdab in 4 hours.
VP Reagent A: Alpha napthol in absolute methyl alcohol VP Reagent B: 40% KOH with 0.3% creatine
POSITIVE
NEGATIVE
Red Ring
Yellow/Brown Ring
Red solution
Yellow solution
Pink to Red color
Yellow or copper-like
MEDIA
PRINCIPLE
FORMATION
MEDIUM
INDICATOR
POSITIVE
NEGATIVE
Citrate Utilization Test
The organism utilizes the citrate (sole src. of carbon) producing ammonia and converted to ammonium hydroxide. This alkaline compound raises the pH of the medium and takes the blue color.
Slant
SCA (Simmon Citrate Agar)
Bromthymol blue
Prussian blue color
Green color
Urease Test
Urease splits the urea molecules into ammonia, carbon dioxide and water. Ammonia reacts in solution to form alkaline compound, ammonium carbonate, which results in an increased pH of the medium and a color change in the indicator to pink-red.
Butt (Broth)
Urea broth
Phenol red
Pink-red
No color change
Sulfide indole motility (SIM)
Butt (Semisolid)
H2S: ferric ammonium chitrate; sodium thiosulfate
Motile Nonmotile (growth (growth pattern – pattern is away from confined) inoculation line) H2S production (Blackening of the butt) Indol production (red ring)
Water Analysis Testing >sanitary supply of water >suitability for general use Method: Multiple Fermentation Tube Technique
Brilliant Green Bile Lactose Broth
Uninocculated
PRESUMPTIVE TEST: LACTOSE BROTH CONFIRMED TEST: BGBL, EMB, ENDO AGAR COMPLETED TEST: LACTOSE BROTH
Positive E. coli + Gas Production + Turbidity...