Lab Practical I

Front 1

• Gram Positive Cocci: Singles pairs, clusters
• SBA: medium, creamy butter, B-hemolytic
• Grows on CNA
• MSA: +
• Catalase: +
• Coagulase (Slide/tube): +

Back 1

Staphylococcus aureus

Front 2

• Gram Positive cocci: singles pairs and clusters
• Grows on SBA (non-hemolytic), CNA
• MSA: =
• Catalase +
• Coagulase (slide/tube): =
• Novobiocin: sensitive >16mm
  • Bacitracin: resistant <10mm

Back 2

Staphylococcus epidermidis

Front 3

• Gram positive cocci: single pairs and clusters
• SBA: non-hemolytic, CNA
• MSA + (yellow)
• Catalase +
• Coagulase (slide/tube): =
  • Novobiocin: Resistant <16mm

Back 3

Staphylococcus saprophyticus

Front 4

• Large gram positive cocci
• small yellow round: SBA, CNA
• MSA: no growth
• Catalas: +
• Coagulase (slide/tube): =
  • bacitracin: sensitive >10mm

Back 4

Micrococcus luteus

Front 5

• Gram positive cocci: single and chains
• SBA/CNA: small to medium grey B-hemolytic
• Catalase: =
• Bacitracin: sensitive
• Hippurate hydrolysis: =
• CAMP test: =
  • PYR: +

Back 5

Streptococcus pyogenes

Front 6

• Gram positive cocci: single and chains
• SBA/CNA: small to medium grey; beta-hemolytic
• Catalase: =
• Bacitracin: Resistant
• Hippurate hydrolysis: Positive
• CAMP test: Positive
  • PYR test: negative

Back 6

Streptococcus agalactiae

Front 7

• Gram positive cocci: single and chains
• SBA/CNA: small grey-white non-hemolytic
• Catalase: negative
• Bile esculin hydrolysis: +
• 6.5% NaCl tolerance: =
  • PYR test: =

Back 7

Streptococcus gallolyticus

Front 8

• Gram positive cocci single and chains
• SBA/CNA: medium grey non-hemolytic
• Catalase: Negative
• Bile esculin hydrolysis: Positive
• 6.5% NaCl tolerance: positive
  • PYR test: positive

Back 8

Enterococcus fecalis

Front 9

• Gram positive cocci: single, pairs, and chains
• SBA: sm-med round dark grey alpha-hemolytic
• CNA: dark grey alpha hemolytic
• Catalase: negative
• Optochin sensitivity: sensitive
  • Bile solubility: positive; colony disappears

Back 9

Streptococcus pneumoniae

Front 10

• Gram positive cocci: single and chains
• SBA: pinpoint small, round light grey alpha-hemolytic
• CNA: light grey alpha-hemolytic
• Catalase: negative
• Optochin: resistant
  • Bile solubility: Negative: colony remains

Back 10

Streptococcus sanguinus (Viridans group)

Front 11

• Gram negative diplococci
• SBA: no growth
• MTM/CA: medium grey colonies
• Oxidase: +
• CTA glucose/maltose/lactose/sucrose:+/-/-/-
  • Butyrate esterase disk: -

Back 11

Neisseria gonorrhoeae

Front 12

• Gram negative diplococci
• SBA/CA/MTM: Medium grey colonies
• Oxidase: +
• CTA glucose/maltose/lactose/sucrose: +/+/-/-
  • Butyrate esterase disk: -

Back 12

Neisseria meningitidis

Front 13

• Gram negative diplococci
• SBA/CA/MTM: medium grey colonies
• Oxidase: +
• CTA glucose/maltose/lactose/sucrose: +/+/+/-
  • Butyrate esterase disk: -

Back 13

Neisseria lactamica

Front 14

• Gram negative diplococci
• SBA/CA(yellow colonies): small grey colonies
• MTM: no growth
• Oxidase: +
• CTA glucose/maltose/lactose/sucrose: +/+/-/+
  • Butyrate esterase: -

Back 14

Neisseria sicca

Front 15

• Gram negative diplococci (larger)
• SBA/CA: small creamy grey colonies
• MTM: no growth
• Oxidase: positive
• CTA glucose/maltose/lactose/sucrose: -/-/-/-
  • Butyrate esterase: positive

Back 15

Moraxella catarrhalis

Front 16

• Small pleomorphic GPR
• SBA/TSA/CNA: small creamy white beta hemolytic
• CTB: black colonies at 24 hrs
• Catalase: +
• Motility: -
• Nitrate:+
• Urea: -
  • Sucrose: -

Back 16

Corynebacterium diphtheriae

Front 17

• Small pleomorphic GPR
• SBA/TSA/CNA: Chalky white pinpoint colonies
• CTB: growth, white colonies at 24hrs, some turn black at 48
• Catalase: +
• Motility: -
• Nitrate: -
  • Urea: -

Back 17

Corynebacterium jeikeium

Front 18

• Small pleomorphic GPR (chinese letters)
• SBA/TSA/CNA: pinpoint colonies
• CTB: growth, white colonies at 24 hours (some turn black at 48)
• Catalase: +
• Motility: -
• Nitrate: negative (should be +)
  • Urea +

Back 18

Corynebacterium pseudodiphtheriticum

Front 19

• Larger GPR, box car appearance
• SBA/TSA/CNA: large ground glass; beta hemolytic grey
• CTB: weak growth
• Catalase: +
• Weak motility

Back 19

Bacillus cereus

Front 20

• Small GPR
• SBA/TSA/CNA: grey beta hemolytic
• CTB; no growth
• Catalase: +
• Motility: umbrella at 25C, motile at 35C
• Bile esculin: +
  • Sucrose: +

Back 20

Listeria monocytogenes

Front 21

• Small GPR
• SBA/TSA/CNA: alpha hemolytic grey
• Catalase: -
• Motility: -
  • TSI slant: K/K H2S

Back 21

Erysipelothrix rhusiopathiae

Front 22

• Small GPR
• SBA: pinpoint growth alpha hemolytic (green agar)
• TSA/CNA: No growth
• Catalase: -
  • Motility: -

Back 22

Lactobacillus

Front 23

• Small gram negative coccobacilli
• CA: grey wet round medium size
• SBA/MAC: no growth
• Satellitism SBA: positive
• X/V: no growth
• Growth between X and V
  • ALA-porphoryn reaction: negative

Back 23

Haemophilus influenzae

Front 24

• Small gram negative coccobacilli
• CA: grey small round
• SBA/MAC: no growth
• X: no growth
• V: growth
• Growth between V and X and all around V
  • ALA-porphoryn reaction: positive

Back 24

Haemophilus parainfluenzae

Front 25

• Small gram negative rods
• CA/SBA: medium sized white/creamy
• MAC: growth at 24hr, more apparent 48hr
• Oxidase: +
• K/K
• Urea: rapid 4 hours
  • Motility: +

Back 25

Bordetella bronchiseptica

Front 26

• Short/fat gram negative rods
• CA/SBA: medium white/creamy
• MAC: no growth
• Oxidase: +
• TSI: A/A
• Urea: -
• Motility: -
  • Spot indole: +

Back 26

Pasteurella multocida

Front 27

• Mac: LF
• TSI: A/A gas
• IMVC:+/+/-/-
• Motile
• L/O/A: +/+/-
• Urea: -
  • Malonate: -

Back 27

Escherichia coli

Front 28

• MAC: LF
• TSI: A/A gas
• IMVC:-/-/+/+
• Non-motile
• L/O/A: +/-/-
• Urea: +
  • Malonate: +

Back 28

Klebsiella pneumoniae

Front 29

• MAC: LF
• TSI: A/A gas
• IMVC: -/-/+/+
• Motile
• L/O/A: +/+/-
• Urea: -
  • Malonate: +

Back 29

Enterococcus aerogenes

Front 30

• NLF
• A/A
• IMVC: +/+/-/-
• Motile at 25, Nonmotile 35
• L/O/A: -/+/-
• Urea: +
  • Malonate: -

Back 30

Yersinia enterocolitica

Front 31

• MAC: NLF
• TSI: K/A H2S gas
• Indole: neg
• Citrate: +
• Motile: +
• ODC: +
• PD: +
  • Urea: +

Back 31

Proteus mirabilis

Front 32

• MAC: NLF
• TSI: A/A H2S gas
• Indole: +
• Citrate: -
• Motile: +
• ODC: -
• PD: +
  • Urea: +

Back 32

Proteus vulgaris

Front 33

• MAC: NLF
• TSI: K/A gas
• Indole: +
• Citrate: -
• Motile: +
  • ODC: +
• PD: +
  • Urea: +

Back 33

Morganella morganii

Front 34

• MAC: NLF
• TSI: K/A
• Indole: +
• Citrate: +
• Motile: +
• ODC: -
• PD: +
  • Urea: +
    • Adonitol/Inositol: +

Back 34

Providencia rettgeri

Front 35

• MAC: NLF
• TSI: K/A (or A/A)
• Indole: +
• Citrate: +
• Motile: +
• ODC: -
• PD: +
  • Urea: -
    • Adonitol/Inositol: -/+

Back 35

Providencia stuartii

Front 36

• MAC: late LF
• TSI: A/A
• IMVC: -/-/+/+
• Motile at 35
• LDC: +
• Malonate: -
• ONPG: +
• L-arabinose: -
  • Dnase: +

Back 36

Serratia marcescens

Front 37

• MAC: weak LF
• TSI: A/A
• IMVC: -/-/+/+
• Motile at 35
• LDC: V (-)
• Malonate: +
• ONPG: +
• L-arabinose: +
  • Dnase: +

Back 37

Serratia rubidaea

Front 38

• MAC: late LF
• TSI: K/A
• IMVC: +/+/-/+
• Motile at 35
• LDC: -
• ODC: +
• Malonate: +
• ONPG: +
• L-arabinose: +
  • Dnase: -

Back 38

Citrobacter diversus

Front 39

• MAC: late LF
• TSI: A/A H2S
• IMVC: -/+/-/+
• Motile at 35
• LDC: -
• ODC: -
• Malonate: -
• ONPG: +
• L-arabinose: +
  • Dnase: -

Back 39

Citrobacter freundii

Front 40

• MAC: NLF
• TSI: K/A H2S gas
• IMVC: -/+/-/+
• Motile at 35
• LDC/ODC/ADC: +/+/+
• PD: -
• Urea: -
• Malonate: -
  • ONPG: -

Back 40

Salmonella Typhimurium

Front 41

• MAC: NLF
• TSI: K/A
• IMVC: -/+/-/-
• Non-Motile at 35
• LDC/ODC/ADC: -/+/-
• PD: -
• Urea: -
• Malonate: -
  • ONPG: +

Back 41

Shigella sonnei

Front 42

• MAC: NLF
• TSI: K/A
• IMVC: -/+/-/-
• Non-Motile at 35
• LDC/ODC/ADC: -/-/-
• PD: -
• Urea: -
• Malonate: -
  • ONPG: -

Back 42

S. flexneri

Front 43

• MAC: NLF
• TSI: K/A weak H2S moustache
• IMVC: -/+/-/-
• Motile at 35
• LDC/ODC/ADC: +/-/-
• PD: -
• Urea: -
• Malonate: -
  • ONPG: -

Back 43

Salmonella Typhi

Front 44

Back 44

1. Staphylococcus aureus
   1. Staphylococcus epidermidis

Front 45

Back 45

Top: Positive; S. aureus

Bottom: Negative; S. epidermidis, S. saprophyticus, M. luteus

Enzyme clots plasma

Cell bound: slide

Extracellular free coagulase: test

Front 46

Back 46

Left: Positive

Right: Negative

2H20--->2H20+O2

Front 47

Produces bound coagulase but not free coagulase

Back 47

Staphylococcus lugdunensis

Front 48

Bacitracin

Back 48

Left: Susceptible >10mm M. luteus, S. pyogenes

Right: Resistant <10mm S. agalactiae

Front 49

Back 49

Left: susceptible >16mm

Right: resistant <16mm

Front 50

MSA

Back 50

• Left: Ferments Mannitol S. saprophyticus, S. aureus
• Right: Does not ferment mannitol S.epidermidis
• M. luteus: does not grow
• Selective for Gram positive due to high salt content
  • Differential for mannitol fermentation

Front 51

PYR

Back 51

• Left: positive Streptococcus pyogenes
  • Right: negative Streptococcus agalactiae

Front 52

CAMP test

Back 52

A: Positive S. agalactiae

B: Negative: S. pyogenes

C: S. aureus

Front 53

Sodium Hippurate

Back 53

Left: Positive; S. agalactiae

Right: Negative S. pyogenes

Ninhydrin added

Front 54

SXT

Back 54

Group C streptococci SXT susceptible

S. pyogenes: SXT disk resistant

Front 55

Back 55

Left: Positive; Enterococcus fecalis

Right: negative

Front 56

6.5% NaCl

Back 56

Cloudy: Positive; E. fecalis

Front 57

Bile solubility

Back 57

• S. pneumoniae; clearing

Front 58

Optochin

Back 58

• Left: Resistant S. sanguinus (Viridans Group) <14mm
• Right: Sensitive S. pneumoniae >14mm

Front 59

Chocolate Agar

Back 59

Enriched growth medium

Contained lysed RBC which has: X and V

Front 60

Back 60

phenol red indicator

If sugar is oxidized; yellow

Front 61

Back 61

A: Positive

B: Negative

Presence of enzyme cytochrome oxidase

Front 62

MTM; modified thayer martin

Back 62

Selective for N. gonorrhoeae, N. meningitidis

Colistin: Inhibits gram negative bacteria other than Neisseria

Vancomycin: inhibit most gram positive bacteria

Nystatin or anisomycin: inhibit yeast

Trimethoprim: inhibit Proteus

Front 63

Nitrate reduction

Back 63

NO3-->NO2-->N2

Red after addition of alpha-naphthylamine, sulfanic acid

Zn detects nitrate not reduce

Red is positive

Front 64

Urease

Back 64

Left: negative acidic

Right: Positive alkaline

Produce urease (hydrolysesammonia)

Front 65

Back 65

carbohydrate fermentor (lactose (1%), glucose (0.1%), sucrose(1%))

• K/K: nonfermentor
• K/A: ferments glucose, and either lactose or sucrose or both
• A/A: ferments glucose and either lactose or sucrose or both
  • H2S: black precip and ferments glucose

Front 66

Eosin Methylene Blue

Back 66

• Selective: Eosin and Methylene blue inhibit GP bacteria
  • Differential: Lactose Fermentation (Dark Pink/Almost Black)

Front 67

MacConkey

Back 67

• Selective: Bile salts and crystal salts- inhibit
  • Differential: fermenting lactose

Front 68

Indole

Back 68

• Measure organisms ability to metabolize tryptophan with the enzyme tryptophanase
• Enzyme degradation leads to formation of pyruvic acid, indole, and ammonia
  • Koch's reagent

Front 69

Back 69

• Mixed acid pathway, pH remains under 4.4 for at least 48 hours
  • Add methyl red

Front 70

Voges Proskauer

Back 70

• Glucose fermented by butylene glycol pathway
• Main end produce: acetylmethylcarbinol
• Addition of alpha-naphthol
• 40% KOH

Front 71

Back 71

• Sodium citrate as the sole source of carbon
  • pH: indicator bromothymol blue turn blue with alkaline pH

Front 72

Bordet-Gengou Agar

Back 72

• Potato starch, glycerol and blood
• Buffered charcoal
• Yeast extract
• Isolates legionella
  • Contains cysteine

Front 73

CA

Back 73

• Can contain: X, V, isovitaleX
  • Glucose, cysteine

Front 74

Malonate

Back 74

• Sodium malonate as a carbon source
  • Bromothymol blue; alkaline product

Front 75

Phenylalanine Deaminase

Back 75

• Enzyme produce phenylalanine deaminase
• Forms pyruvic acid will form a dark green complex
  • FeCl3

Front 76

Shigella Serotypes

Back 76

• A: S. dysenteriae
• B: S. flexneri
• C: S. bodyii
  • D: S. sonnei

Front 77

Hektoen Enteric Agar (HE)

Back 77

• Selective: Bile salts
• Ferric ammonium citrate and sodium thiosulfate: Detects H2S
• Indicator: Acid fuchsin and bromothymol blue
  • Green colonies: don't ferment lactose

Front 78

Xylose Lysine Desoxycholate (XLD)

Back 78

• Selective: sodium deoxycholate; inhibits gram positive
• Shigella does not ferment lactose
• Differental: lactose; alkaline pH
  • H2S: sodium thiosulfate ferric ammonium citrate

Front 79

Cefsulodin-Irgasan-Novobiocin (CIN)

Back 79

• Selective inhibition: Crystal violet, sodium desoxycholate and Irgasan
• Supplementation: cefsulodin (inhibits normal enteric organisms)
  • For isolation of Yersinia entercolitica

Front 80

Fermentation of L-arabinose

Back 80

S. marcescens is the only Enterobacteriaceae that does not ferment L-arabinose

Pos: Yellow

Front 81

ONPG

Back 81

Differentiates between lactose and non-lactose fermentors

ONPG: is an analog of Lactose

Hydrolzyed by Beta-galactosidase

Front 82

DNase

Back 82

Dnase degrades extracellular DNA

Serratia: +

Enterobacteriaceae: =