Sunday, December 9, 2007

Group Post: MMIC dPBL-1

Biochemical and Culture Testing


CASE 1

Possible Organisms Biochemical Tests & Results Culture agars
Escherichia coli
Gram Stain: -
Oxidase: -
Lactose: +
Lysine: +
TSI slant: +
Indole: +
Methyl red: +
VP: -
Citrate: -

5% Sheep Blood Agar
Eosin-Methylthionine Blue Agar
MacConkey Agar
Pseudomonas aeruginosa
Gram stain: -
Oxidase: +
Pyocyanin: +
Fluoreseein: +
Citrate: +
Nitrase: +
Lipase: +
Ureases: +/-
Indole: -
TSI: -

Pseudomonas Agar P
Pseudomonas Agar F
MacConkey agar plate
Eosin-Methylthionine Blue agar
Cetrimide agar
Staphylococcus aureus
Gram Stain: +
Catalase: +
Coagulase: +

5% Sheep Blood Agar
Mannitol Salt Agar
Staphylococcus saprophyicus
Gram Stain: +
Catalase: +
Coagulase: -
Oxidase: -
Phosphatase: -
Urease: +
Lipase: +
2-hour PYR broth hydrolysis: -
D-trehalose; +

5% Sheep Blood Agar
Mueller-Hinton Agar Lawn with Urine
Enterococcus faecalis
Gram Stain: +
Catalase: -

5% Sheep Blood Agar
Chocolate Agar
6.5% NaCl Concentration Agar (halotolerant)
Bile Esculin Agar (BEA) Slants
Proteus-Providencia-Morganella
e.g. Proteus mirabilis

Gram Stain: -
Lactose: -
Oxidase: -
Urease: +
Indole: -
Nitrogen Reductase: +
Methyl Red: +
Vogues-Proskauer: -
Catalase: +
Phenylalanine Deaminase: +

MacConkey Agar
CLED
Klebsiella-Enterobacter-Serratia
e.g. Klebsiella oxytoca

Gram Stain: -
Oxidase: -
Lactose: -
Urease: -
Nitrate: +
Indole: +
Sucrose: +
Dextrose: -
Methyl Red: -
Vogues-Proskauer: +
Gelatin: -
Amylase: +
Lipase: -
Catalase: +
Citrate: +
H2S: -

5% Sheep Blood Agar
MacConkey Agar
Trypticase Soy Agar
Klebsiella-Enterobacter-Serratia
e.g. Klesiella pneumoniae

Indole: -
Sucrose: +
Dextrose: -
Methyl Red: -
Vogues-Proskauer: +
H2S: -
Lysine Decarboxylase: +

5% Sheep Blood Agar
MacConkey Agar

Done by Lizzie


CASE 2

Campylobacter jejuni
Oxidase (+)
Sensitivity to nalidixic acid

Blood agar with antibiotics to other fecal microbes
Fail to grow at 25C, must grow at 42C
Microaerophilic (grow at 5% oxygen + 10% carbon dioxide)
Shigella species
TSI (alkaline slant and acidic butt, with no gas and no H2S)

MacConkey's or EMB agar (colourless colonies)
Anaerobe
Escherichia coli
TSI (acidic slant and acidic butt, with gas ,but no H2S)
Indole test (+)

MacConkey's (pink colonies) or EMB agar (green sheen colonies)
Anaerobe
Clostridium perfringens
ELISA to detect Clostridium perfringens Enterotoxin

TSC agar containing egg yolk
Neomycin blood agar
Strict anaerobe
Clostridium difficile
Cytotoxicity test on human cell-line
ELISA to detect Clostridium perfringens exotoxin

Cycloserine cefoxitin fructose egg yolk agar (CCFA)
Vibrio Cholerae
Oxidase (+)
TSI (acidic slant acidic deep; no gas)
Agglutination of bacteria by polyvalent O1 antibody

TCBS (yellow colonies), Blood agar
Vibro parahemolyticus
Oxidase-positive
Grow in 10% NaCl

TCBS (green colonies)
Bacillus cereus
Lecithinase (+)
Motile (swarming on agar)
Catalase (+)
Indole (-)
Nitrate (+)
Urease (-)
Citrate (+)
MR (-)
VP(+)
TSI (Alkaline slant, acidic deep)
Oxidase (-)
Lactose fermenting; sucrose non-fermenting

Sheep’s blood Agar (beta hemolytic), large greenish colonies),
Mannitol (grey colonies)
Yersinia enterocolitica
Motile at 25 deg
MR (+)
VP (-)
Urease (-)
Indole (-)
Citrate (-)
Catalase (+)
TSI (acidic slant acidic deep; no gas)

MacConkey Agar (small lactose negative colonies)
EMB
Staphylococci aureus
Catalase (+)
Citrate (-)
Coagulase (+)
Indole (-)ME (+)
Oxidase (-)
Urease (-)
VP (-)
TSI (acidic slant acidic deep)

Sheep blood agar (beta hemolytic, yellow colonies)
Mannitol salt agar (yellow colonies)
7.5% NaCl media

Done by Kent and Ye Tun


CASE 3

E. coli Oxidase -
Catalase +
Indole +
Methyl red +
Voges-Prokauer -
Citrate Utilization -
Urease -
TSI; H2S production Acid slant/ acid butt & gas produced;
+
MacConkey agar Lactose fermenters; (red colonies)
Blood agar β-hemolytic
Klebsiella sp (Klebsiella pneumuniae, Klebsiella oxytoca) Oxidase -
Catalase +
Indole +
Methyl red -
Voges-Prokauer +
Citrate Utilization +
Urease -
TSI; H2S production Alkaline slant/ acid butt;
-
MacConkey agar Lactose fermenters; (pink colonies)
Blood agar Mucoid colonies formed
Enterobacter (Enterobacter aerogenes) Oxidase -
Catalase +
Indole -
Methyl red -
Voges-Prokauer +
Citrate Utilization +
Urease
TSI; H2S production
MacConkey agar
Blood agar
Serratia Oxidase -
Catalase +
Indole
Methyl red -
Voges-Prokauer +
Citrate Utilization +
Urease
TSI; H2S production
MacConkey agar
Blood agar
Proteus mirabilis Oxidase -
Catalase +
Indole +
Methyl red +
Voges-Prokauer -
Citrate Utilization
Urease +
TSI; H2S production Alkaline slant/ acid butt;
+
MacConkey agar Non-lactose fermenters; (white colonies)
Blood agar Swarming observed
Morganella morganii Oxidase -
Catalase +
Indole
Methyl red
Voges-Prokauer
Citrate Utilization
Urease +
TSI; H2S production
MacConkey agar Non-lactose fermenters; (white colonies)
Blood agar
Pseudomona aeruginosa Oxidase +
Catalase +
Indole -
Methyl red
Voges-Prokauer
Citrate Utilization +
Urease
Coagulase
TSI; H2S production
MacConkey agar Non-lactose fermenters; (white colonies)
Blood agar
Staphylococcus aureus
Catalase +
Methyl red
Voges-Prokauer
Citrate Utilization
Coagulase +
MacConkey agar -
Blood agar β-hemolytic
Staphylococcus saprophyticus
Catalase +
Methyl red
Voges-Prokauer
Citrate Utilization
Urease +
Coagulase -
MacConkey agar Non-lactose fermenters; (white colonies)
Blood agar -
Enterococcus sp
Catalase
Indole
Methyl red
Voges-Prokauer
Citrate Utilization
Urease
Coagulase
TSI; H2S production
MacConkey agar
Blood agar

Done by Joan


CASE 4

Possible Organisms Biochemical Tests & Results Culture agars
Streptococcus pneumoniae
Gram Staining +ve
Catalase -ve

2. 5% Sheep Blood agar with Optocin sensitivity Alpha-hemolysis with sensitivity to optocin
Klebsiella-Enterobacter-Serratia (Klebsiella sp.)
Gram Staining -ve
Oxidase -ve
Indole +ve/-ve
MR-VP +/- or -/+
Citrate +ve
H2S Production -ve
Lysine Decarbocylase +ve

Lactose Fermentation (MacConkey agar) +ve
Bordetella pertussis
Gram Staining -ve
Oxidase +ve
Urease -ve
Nitrase -ve
Citrate -ve

Borde+ - Gengou agar
BCYE agar

Done by Charmaine


CASE 5

Staphylococcus aureus
Gram Staining +
Catalase +
Coagulase +

Mannitol Salt Agar with 7-9% NaCl Mannitol fermentation
Sheep Blood Agar Hemolysis of blood agar
Streptococcus pyogenes
Gram Staining +
Catalase -
PYR (Pyrrolidonearylamidase) Test +

Sheep Blood Agar Beta Hemolysis
Enterococci
Gram Staining +
Bile- esculin test -

Sheep Blood Agar Gamma Hemolysis
6.5% NaCl Agar Can grow with 6.5% Nacl
Escherichia coli
Gram Staining -
Indole +
Methyl Red +
Voges Prokauer -
Citrate -
TSI Acid slant and butt with gas production but no H2S in TSI

MacConkey Lactose fermentative (deep red colony)
Eosin Methylene Blue( EMB) Green metallic sheen on EMB
Pseudomonas aeruginosa
Gram Staining -
Oxidase +
Catalase +
Indole +
Citrate +
Urease -
TSI -
Pyocyanin -
Fluroscein -

MAC Non- lactose fermentative (colourless colony on MAC)
Pseudo F agar PositiveFluorescein pigment in Pseudo F agar

Done by Ai Tee


CASE 6
Gardnerella vaginalis
Catalase -
Oxidase -
Vaginal pH test pH of vaginal fluid >5.0
Whiff test Release of bad-smelling odor when discharge is mixed with KOH
DNA Probe (Affirm™ VP III) High concentrations of G. vaginalis in VP III test

Chocolate agar Small, circular, convex, gray colonies
HBT agar
Colistin-oxolinic acid blood agar Beta-hemolysis
E. coli
Triple Sugar Iron (TSI) test TSI test show acid slant, acid butt with gas.
Indole test +
Methyl red test +
Voges-Proskauer (VP) test -
Citrate test -

EMB agar Black colonies with greenish-black metallic sheen on EMB agar
MacConkey agar Deep red colonies on MacConkey agar (lactose fermentor)
TSI agar
Pseudomonas aeruginosa
Oxidase test +
Catalase test +
Nitrase tese +
Lipase test +
TSI test Metallic sheen growth on the surface of TSI agar

EMB agar
Mac Conkey’s agar (sterile specimen)
Cetrimide agar (non-sterile specimen) Colonies appear flat, large, and oval. Secretes blue-green pigment (pyocyanin) on Cetrimide agar
TSI agar No colour change on TSI medium (K/K/g-/H2S- profile)
Neisseria gonorrhoeae
Oxidase test +
Direct immunofluorescence

Thayer-Martin chocolate agar Growth of spherical shape colonies on TM chocolate agar
Chlamydia Trachomatis
Direct immunofluorescence Presence of monoclonal antibodies (mAbs) to epitopes in the VS4 region of MOMP
ELISA Presence of Chlamydia antigens in ELISA test
PCR
DNA-based tests.

Blood agar
Mac Conkey’s agar
Trichomonas vaginalis
Vaginal pH test pH of vaginal fluid >4.5
PCR
Enzyme immunoassay Presence of T. vaginalis antigen in enzyme immunoassay
Direct Immunofluorescence.

Blood agar
Mac Conkey’s agar
Mobiluncus
Catalase test -
Indole test -
Oxidase test -

Blood agar Clear, colourless colonies, around 2mm.
Mac Conkey’s agar
Enterobacter-Klebsiella-Serrtia
-

Blood agar
Mac Conkey’s agar Deep red colonies on MacConkey agar (lactose fermentor)
Proteus-Providencia-Morganella
Urease test +
Indole test +

Blood agar Appear as swarming on blood agar.
Mac Conkey agar Colourless colonies on MacConkey agar (Non-lactose fermentor)
Enterococcus faecalis
Catalase test -

Blood agar Alpha, beta, or non-hemolytic on blood agar.
Mac Conkey agar
Candida albicans
-

Blood agar
Sabouraud’s agar Presence of white, butyrous colonies.
Staphylococcus saprophyticus
Catalase test +
Coagulase test -
Urease test +
Lipase test +

Blood agar
Mac Conkey’s agar Spherical, irregular grape-like cluster in culture.

Done by Adrian

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