Bacteriology

Front 1

Tiny satellite colony around Staphaureus

Back 1

Haemophilus Infleunza

Front 2

Haemophilus Species

Back 2

H. Infleunzae

H. Parainfleunzae

H. Ducreyi

Front 3

Haemophilus (Specimen Requirements)

Back 3

1-General principles

2-Susceptible to drying and extreme temperatures -> Immediate plating

3-Special Measurements for H. Ducreyi recover from genital ulcers

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Haemophilus (General Characteristics)

Back 4

Gram -

Coccobacilli

Oxidase Variable

Catalase +

Non-pigmented

Slightly yellowish

Ultrastructural features similar to other pathogenic organisms

All members are aerobic or facultative anaerobic

Fastidious organisms

Cannot grow on MacConkey

Front 5

X and V Growth Requirement Test

Back 5

Principle

1- X and V factor disks are paper disks impregnated with X (Hemin) and V (NAD: Nicotinamide Adenine Dinucleotide) growth factors

2. Ddifferentiate Haemophilus species based on their requirements for these growth factors

Procedure

1- Inoculate Muller Hinton agar with the bacterial suspension using a sterile swab to obtain confluent growth

2- Aseptically apply one X factor disk, one V factor disk, and one XV factor disk and slightly press down to ensure full contact with the medium

3- Incubate for 18-24 hrs at 35-37C in 3-5% CO2

4- Examin for growth around the disks

Results:

1- Growth around X and XV disk factors --> factor X is required by the organism

2- Growth around V and XV disk factors --> factor V is required by the organism

3- Growth around XV factor only --> factorsnX and V are required by the organism to grow

Precautions:

1- Do not use blood and chocolate agar

2- Blood has X factor (Hemin) and Chocolate has X and V factors (Hemin and NAD: Nicotinamide Adenine Dinucleotide)

3- Give false positive results

Bacteria:

Haemophilus

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Haemophilus (Anitmicrobial Susceptibility Testing)

Back 6

According to CLSI:

1- Use HTM ( Haemophilus Test Medium) instead of Muller Hinton Agar

2- Has the necessary nutrients and growth factors required for these fastidious organisms' confleunt growth

Front 7

Identify

Back 7

Haemophilus Species

1- Non-pigmented

2- Slightly Yellowish in color

Front 8

Horse blood- bacitricin Agar

Back 8

1- Solid Medium

2- Quantitative procedures

3- Selective isolation

4- Haemophilus Species (H. Infleunzae)

5- Respiratory clinical specimen

Front 9

Identify

Back 9

Horse-blood-bacitricin Agar

1- Solid Medium2- Quantitative procedures3- Selective isolation4- Haemophilus Species (H. Infleunzae)5- Respiratory clinical specimen

Front 10

Beta-Lactamase Detection Test

Back 10

Principle:

1- Determines the ability of some bacteria to produce beta lactamase enzyme that hydrolyze the beta lactam-ring component destroying the antimicrobial activity of some beta-lactam drugs.

2- Chromogenic Cephalosporins (beta-lactam antimicrobial agent) changes the color from yellow to pink) when their beta-lactam ring has been hydrolyzed by the beta lactamase enzyme produced by the tested organism.

Procedure:

1- Place a beta-lactam disk on a clean glass slide.

2- Hydrate it by a dropper and distilled water

3- Rub the colony to be tested on the hydrated disk by a wooden stick

4- Examin Change in color in the inoculated area within 5s-10min

Results

1- Deep Pink Color --> +

2- Yellow Color (no change in color) --> -

Bacteria:

1-Haemophilus Infleunzea +

Front 11

Identify:

Back 11

Beta lactamase test

Principle:1- Determines the ability of some bacteria to produce beta lactamase enzyme that hydrolyze the beta lactam-ring component destroying the antimicrobial activity of some beta-lactam drugs.2- Chromogenic Cephalosporins (beta-lactam antimicrobial agent) changes the color from yellow to pink) when their beta-lactam ring has been hydrolyzed by the beta lactamase enzyme produced by the tested organism.Procedure:1- Place a beta-lactam disk on a clean glass slide.2- Hydrate it by a dropper and distilled water3- Rub the colony to be tested on the hydrated disk by a wooden stick4- Examin Change in color in the inoculated area within 5s-10minResults1- Deep Pink Color --> +2- Yellow Color (no change in color) --> -Bacteria:1-Haemophilus Infleunzea +

Front 12

Identify (test)

Back 12

X and V Growth Requirements Tests

Principle1- X and V factor disks are paper disks impregnated with X (Hemin) and V (NAD: Nicotinamide Adenine Dinucleotide) growth factors2. Ddifferentiate Haemophilus species based on their requirements for these growth factorsProcedure1- Inoculate Muller Hinton agar with the bacterial suspension using a sterile swab to obtain confluent growth2- Aseptically apply one X factor disk, one V factor disk, and one XV factor disk and slightly press down to ensure full contact with the medium3- Incubate for 18-24 hrs at 35-37C in 3-5% CO24- Examin for growth around the disksResults:1- Growth around X and XV disk factors --> factor X is required by the organism2- Growth around V and XV disk factors --> factor V is required by the organism3- Growth around XV factor only --> factorsnX and V are required by the organism to growPrecautions:1- Do not use blood and chocolate agar2- Blood has X factor (Hemin) and Chocolate has X and V factors (Hemin and NAD: Nicotinamide Adenine Dinucleotide)3- Give false positive results Bacteria:Haemophilus

Front 13

Identify

Back 13

Pleomorphic Gram - Coccobacilli Haemophilus

Front 14

Identify

Back 14

1- X factor is required for growth --> H. Ducreyi

Front 15

Identify

Back 15

H. Infleunzae

1- Requires both X and V factors

2- Beta lactamase positive

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H. Infleunza

Back 16

1- Gram negative coccobacillin (on chocolate agar)

2- Factor X/ Factor V/ Factor XV : -/-/+

3- Beta Lactamase Positive

Front 17

Tests

1- Gram negative coccobacillin (on chocolate agar)

2- Factor X/ Factor V/ Factor XV : -/-/+

3- Beta Lactamase Positive

Back 17

H. Infleunzea

Front 18

H. Ducreyi

Back 18

1- Gram negative coccobacillin (on chocolate agar)

2- Factor X/ Factor V/ Factor XV : +/-/+

Front 19

Tests

1- Gram negative coccobacillin (on chocolate agar)

2- Factor X/ Factor V/ Factor XV : +/-/+

Back 19

H. Ducreyi

Front 20

Tests

1- Gram negative coccobacillin (on chocolate agar)

2- Factor X/ Factor V/ Factor XV : -/+/+

Back 20

H. Parainfleunzea

Front 21

Haemophilus Detection

Back 21

1-Direct Detection by light microscopy (pleomorphic gram - coccobacilli)

2-Commercial available latex agglutination kits for the detection of H. Influenzea type b capsular polysaccharide

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Muller-Hinton-based Chocolate Agar

Back 22

1- Enriched Medium

2- Isolation and Cultivation

3- Fastidious Organisms

4- Particularly, H. Ducreyi

5- Supplemented with 1% Isovitalix

6- Supplemented with 3ug/ml Vancomycin for thr inhibitiom of gram + organisms coloniIng the genital tract

Front 23

Heart-Infusion-based Agar

Back 23

1- Enriched Medium

2- Isolation and Cultivation

3- Fastidious Organisms

4- Particularly, H. Ducreyi

5- Supplemented with 10% fetal bovine serum

6- Supplemented with 3ug/ml Vancomycin for thr inhibition of gram + organisms colonizing the genital tract

Front 24

Burkholderia (General Characteristics)

Back 24

1- Previoudly a part of pseudomonas (Genus)

2- Gram -

3- Rod-shaped Bacteria (Bacilli)

4- Oxidase Positive

5- Non-Lactose Fermenter

6- Obligate Aerobic

7- Motile by means of single or multiple polar flagella (except B. Mallei)

8-"Grows on MacConkey

Front 25

Burkholderia (Species)

Back 25

Human pathogen:

1- B. Cepacia (pulmonary infection in patients with cystic fibrosis CF)

2- B. Pseudomalliae

Others

3- B. Malliae

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Burkholderia (Media)

Back 26

Broth

1- Thioglycolate

2- Broth-blood culture systems

Agar

1- Blood Agar

2- Chocolate Agar

3- MacConkey Agar

4- OFPBL: Oxidative-fermentative-polymyxin B-bacitricin- lactose agar

5- PC: Pseudomonas Cepacia agar

7- Ashdown's Agar

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OFPBL: Oxidative-fermentative-polymyxin b-bacitricin-lactose Agar

Back 27

1- Selective

2- Solid Medium

3- Qualitative Procedures

4- Detection and isolation

5- B. Cepecia

6- From clinical speciemem (Respiratory Secretions of CF patients)

6- Colonies: Yellow with yellow halos

Front 28

Pseudomonas Cepacia (CP) Agar

Back 28

1- Selective

2- Differential

3- Detection and isolation

4- B. Cepacia

5- Fromclinical specimen and mixed cultures of CF patients

Contains:

6- Bile salts and crystal violet which inhibit gram + bacteria

7- Polymyxin B and Ticarcillin which inhibut gram - bacteria

8- B. Cepacia utilizes pyruvate present in the medium --> alkaline products changing the medium color from bright pink to red (phenol red is the indicator)

Front 29

Ashdown's Medium Agar

Back 29

1- Selective

2- Differential

3- Isolation and detection

4- B. Pseudomalliae

Contains

5- Crystal violets and gentamicin to inhibit the growth of the other bacteria

6- B. Pseudomalliae take up the neutral red ( in medium) to produce flat wrinkled purple colonies

Front 30

Vibrio (General Characteristics)

Back 30

1- Gram -

2- Rod-shaped (Bacilli)

3- Oxidase +

4- Non-lactose fermenter

5- Glucose fermenter

6- Beta Hemolytic

7- Grow on MacConkey

Front 31

Vibrio (Specimen Requirements)

Back 31

1- General principles

2- Stools samples suspected to contain Vibrio specieaahould be colleted in Cary-blair medium

Front 32

Vibrio (Media)

Back 32

Broth:

Thioglycolate

Broth-blood culture system

Alkaline peptone water broth

Agar:

1- Blood (Beta Hemolytic)

2- Chocolate

3- MacConkey (NLF)

4- TCBS: Thiosulfate citrate bile salts sucrose agar

Front 33

Alkaline Peptone Water Broth

Back 33

1-Enrichment

2- Vibrio species

3- From food, water, and clinical specimen

4- Bacterial sample inoculated, incubated for 5-8 hrs at 35-37C

5- Subcultured in TCBS agar

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TCBS: Thiosulfate citrate bile salts sucrose agar

Back 34

1- Selective Agar

2- Isolation and cultivation

3- Vibrio Species

4- Alkaline pH of the medium enhances the growth of V. Cholerae and inhibits that of others

5- Sucrose (for metabolism) is fermented by the V. Cholerae --> acid end product

6- Colonies yellow and large

7- Bromothymol blue and thymol as indicators

Front 35

Vibrio Cholerae (Direct Detection)

Back 35

1- Microscopy: Gram negative bacilli

2- Serotyping of toxigenic strains of V. Cholerae (01 and 0139) using specific antisera. Negative for both strains is reported as non-01

3- ELISA (Enzyme linked immunosorbent assay) and commercial available latex agglutination kits for the rapid detection of the toxin

Front 36

Aeromonas

Pleisomonas (shigelloides)

Chromobacterium (violaceum)

(General Characteristics)

Back 36

1- Gram -

2- Bacilli

3- Oxidase +

4- NLF

5- Glucose Fermenters

7-Grow on MacConkey

6- Aeromonas/ Chromobacterium beta hemolytic

Pleisomonas non-beta hemolytic

Front 37

Aeromonas

Pleisomonas (shigelloides)

Chromobacterium (violaceum)

(Media)

Back 37

Broth:

1- Thiogylcolate

2- Broth-blood culture systems

Agar:

1- Blood Agar

2- Chocolate Agar

3- Maconkey Agar

4- TSA II: Tryptic Soy Agar with 5% SRBC and ampicillin

Front 38

TSA II: Tryptic Soy Agar with 5% SRBC and ampicillin

Back 38

1- Selective

2- Isolation and cultivation

3- Aeromonas spp

4- Ampicillin inhibits Enterobacteriaceae and some gram positive bacteria

5- Colony: Smooth, Convex, Grayish, Beta-hemolytic

Front 39

String Test

Back 39

1- Differentiate V. Cholerae (+) from Aeromonas spp (-) since both are isolated feom diarrheal stools/ have same biochemical properties on culture media/ oxidase +

2- Differentiate V. Cholerae (+) from othe Cholerae species

3- Isolated colony (18-24hr) from suspected bacterium is emulsified with Na Deoxycholate (known as bile salts) it lyses the cell releases the DNA.

4- Suspension loses its turbidity and becomes viscous

5- Mucoid string is formed when an inoculating loop is drawn slowly away from the suspension

Front 40

Ekinella w atba3oha

Back 40

1- Gram -

2- Bacilli

3- Oxidase +

4- FA

5- Non-motile

6- Fastidious

7- Do not grow on MacConkey

8- Grow on blood and chocolate

9- Pit the agar

Front 41

Ekinella Spp

Back 41

Ekinella Corrodens

1- Gram -

2- Bacilli

3- Oxidase +

4- Nonmotile

5- FA

6- Grow on blood and chocolate

7- Does not grow on MacConkey

8- Takes its name from its ability to corrode (pit) the agar

9- Exude Chlorine bleach odor

10- non-hemolytic

11- Slight greening may occur Round th colony

Front 42

Pasteurella Multocida

Back 42

Pasteurella Multocida

1- Gram - 2- Bacilli 3- Oxidase +4- Nonmotile5- FA6- Grow on blood and chocolate 7- Does not grow on MacConkey

8- Pit the agar

9- Non-hemolytic

11- Translucent

10- Exude must/ mushroom order

Front 43

Actinobacillus (Aggregatibacter) Actinomycetemcomitans

Back 43

Actinobacillus (Aggregabacter) Actinomycetemcomitans

1- Gram - 2- Bacilli 3- Oxidase +4- Nonmotile5- FA6- Grow on blood and chocolate 7- Does not grow on MacConkey

8- Non-hemolytic

9- Translucent

10- pinpointed after 24 hr

11- Slight Tinge after 48 hr

Front 44

Kingella Denitrifcans

Back 44

Kingella Kingae

1-Small

2- Translucent

3- Non-hemolytic

After 24 hr

4- Grow on Thayer-Martin Agar

1- Gram - 2- Bacilli 3- Oxidase +4- Nonmotile5- FA6- Grow on blood and chocolate 7- Does not grow on MacConkey8- Pit the agar

Front 45

Kingella Kingae

Back 45

Kingella Kingae

1- Translucent

2- Or slightly opaque

3 - Beta Hemolytic

1- Gram - 2- Bacilli 3- Oxidase +4- Nonmotile5- FA6- Grow on blood and chocolate 7- Does not grow on MacConkey8- Pit the agar

Front 46

Cardiobacterium Hominis

Back 46

Cardiobacterium Hominis

1- Circular

2- Smooth

3- Opaque

4- Slight alpha hemolytic

After 48 hr

1- Gram - 2- Bacilli 3- Oxidase +4- Nonmotile5- FA6- Grow on blood and chocolate 7- Does not grow on MacConkey8- Pit the agar

Front 47

Capnocytophaga Spp

Back 47

Capnocytophaga Spp

1- yellow

2- Pink

3- nonhemolytic

1- Gram - 2- Bacilli 3- Oxidase +4- Nonmotile5- FA6- Grow on blood and chocolate 7- Does not grow on MacConkey

8- Pit the agar

After 48 hr