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Haemophilus and Other Fastidious GramNegative Rods

INDIAN DENTAL ACADEMY
Leader in continuing dental education
www.indian...
Case Study
 A 2-year-old unvaccinated child was seen in the
emergency room because of complaints of headache
and fever
 ...
Points to Consider
 What clinical findings led the physician to request a
CSF culture?
 Based on the patient’s age, whic...
Haemophilus and Other Fastidious Gramnegative Rods
 The fastidious group of gram-negative bacilli
include:
 Haemophilus
...
Haemophilus Influenzae: Clinical Infections
 Infections caused by typable (encapsulated) strains
 Acute epiglottis or la...
Haemophilus Influenzae: Clinical Infections
 Infections caused by Nontypable strains
 Otitis media
 Sinusitis
 Pneumon...
Haemophilus Species
H. influenzae
satellitism around
and between the
large, white,
hemolytic
staphylococci
Haemophilus spe...
Haemophilus Species

Direct smear of H. influenzae in CSF in a
case of meningitis. Note the intracellular
and extracellula...
Haemophilus species
H.influenzae growing on
chocolate agar. Notice the
semi-opaque, gray-white,
mucoid colonies characteri...
Haemophilus Species: Identification

This organism would be identified as H. influenzae
because it is using both X and V f...
Haemophilus Species: Identification

This organism would be identified as H.
parainfluenzae because it is using V factor o...
Haemophilus Species: Identification

This organism would be identified as H. aphrophilus because it is
using X factor only...
Haemophilus Species: Identification

Under ultraviolet light, the organism on the bottom
is showing a positive porphyrin r...
HACEK Group and Capnocytophaga
 HACEK is an acronym of the first initial of each genus
that belong in the group:
 Haemop...
HACEK Group and Capnocytophaga: General
Characteristics
 Gram-negative bacilli
 Require an increased CO2 (5%-10%) enviro...
HACEK Group
H. aphrophilus growing
on blood agar

Gram stain morphology of H.
aphrophilus
W.B. Saunders Company items and ...
HACEK Group
A. actinomycetemcomitans on
blood agar. Note the starshaped centers of the
colonies.

Microscopic morphology o...
HACEK Group
The 48-hour growth of
colonies of C. hominis

Gram stain of C. hominis
showing typical “rosettes”

W.B. Saunde...
HACEK Group
E. corrodens on
chocolate agar

Gram stain morphology
of E. corrodens

W.B. Saunders Company items and derived...
HACEK Group and Capnocytophaga
Growth of Capnocytophaga
colonies on chocolate agar.
Compare this growth with
Eikenella

Gr...
HACEK group

Gram stain of Kingella kingae showing plump
rods in chains. Compare this morphology with
other members of the...
Pasteurella species
 General characteristics
 Colonizes mucous membranes of the upper respiratory tract
and gastrointest...
Pasteurella species
 P. multocida: Clinical manifestations
 Localized infection after a bite or scratch
 Respiratory tr...
Pasteurella Multocida
 Culture characteristics
 Growth on 5% blood or chocolate shows small, smooth, convex
colonies
 “...
Pasteurella Multocida
P. multocida growing on blood
agar and chocolate agar. No
growth on MacConkey agar
plate.

Gram stai...
Case Study
 Ten members of a group of 2 dozen retirees from the
tobacco industry became acutely ill with pneumonia
during...
Case Study
 On the fifth day of the cruise, several members went
to see the ship’s doctor because of a worsening
cough
 ...
Legionella Species
 General characteristics
 Ubiquitous gram-negative rods
 Acquired by humans primarily through inhala...
Legionella pneumophila
Gram stain of specimen showing
intracellular and extracellular
Legionella pneumophila

L. pneumophi...
Legionella pneumophila

A

B

(A) Nonselective buffered charcoal yeast extract (BCYE) plate
inoculated with sputum specime...
Legionella pneumophila

L. pneumophila colony on BCYE agar after 3, 4, 5, and
7 days of incubation
W.B. Saunders Company i...
Legionella pneumophila: Identification

Schema for identification of L. species

W.B. Saunders Company items and derived i...
Points to Remember
 Clinical manifestations presented by the patient
 The types of infections these organisms produce
 ...
Points to Remember
 What special growth supplements are required for
isolation
 Where these groups of organisms are usua...
Thank you
For more details please visit
www.indiandentalacademy.com

W.B. Saunders Company items and derived items copyrig...
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Haemophilus and other fastidious gram negative rods /certified fixed orthodontic courses by Indian dental academy

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The Indian Dental Academy is the Leader in continuing dental education , training dentists in all aspects of dentistry and offering a wide range of dental certified courses in different formats.

Indian dental academy provides dental crown & Bridge,rotary endodontics,fixed orthodontics,
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Haemophilus and other fastidious gram negative rods /certified fixed orthodontic courses by Indian dental academy

  1. 1. Haemophilus and Other Fastidious GramNegative Rods INDIAN DENTAL ACADEMY Leader in continuing dental education www.indiandentalacademy.com www.indiandentalacademy.com
  2. 2. Case Study  A 2-year-old unvaccinated child was seen in the emergency room because of complaints of headache and fever  A cerebrospinal fluid(CSF) sample was obtained and sent to the laboratory for culture  The Gram stain showed many white blood cells and many gram-negative, small bacilli W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  3. 3. Points to Consider  What clinical findings led the physician to request a CSF culture?  Based on the patient’s age, which organism would be suspected?  What growth requirements must be met to recover the suspected agent?  Which other fastidious organisms would be considered?  Other points to consider W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  4. 4. Haemophilus and Other Fastidious Gramnegative Rods  The fastidious group of gram-negative bacilli include:  Haemophilus  HACEK  Legionella  Bordetella  Pasteurella  Brucella  Francisella W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  5. 5. Haemophilus Influenzae: Clinical Infections  Infections caused by typable (encapsulated) strains  Acute epiglottis or laryngotracheal infection in small children  Cellulitis/arthritis  Meningitis  Pneumonia/septicemia (in children)  Conjunctivitis W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  6. 6. Haemophilus Influenzae: Clinical Infections  Infections caused by Nontypable strains  Otitis media  Sinusitis  Pneumonia, bronchitis (in adults) W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  7. 7. Haemophilus Species H. influenzae satellitism around and between the large, white, hemolytic staphylococci Haemophilus species require hemoglobin for growth: X-factor ( hemin): Heat-stable substance V-factor (NAD): Heat- labile, coenzyme I, nicotinamide adenine dinucleotide, found in blood or secreted by certain organisms W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  8. 8. Haemophilus Species Direct smear of H. influenzae in CSF in a case of meningitis. Note the intracellular and extracellular pleomorphic gramnegative bacilli. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  9. 9. Haemophilus species H.influenzae growing on chocolate agar. Notice the semi-opaque, gray-white, mucoid colonies characteristic of encapsulated strains. Gram stain of H. influenzae from colony W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  10. 10. Haemophilus Species: Identification This organism would be identified as H. influenzae because it is using both X and V factors. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  11. 11. Haemophilus Species: Identification This organism would be identified as H. parainfluenzae because it is using V factor only. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  12. 12. Haemophilus Species: Identification This organism would be identified as H. aphrophilus because it is using X factor only. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  13. 13. Haemophilus Species: Identification Under ultraviolet light, the organism on the bottom is showing a positive porphyrin reaction, whereas the organism on the top is demonstrating a negative porphyrin reaction. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  14. 14. HACEK Group and Capnocytophaga  HACEK is an acronym of the first initial of each genus that belong in the group:  Haemophilus aphrophilus  Actinobacillus actinomycetemcomitans  Cardiobacterium hominis  Eikenella corrodens  Kingella species  Capnocytophaga sp.  Has similar requirements as the HACEK group W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  15. 15. HACEK Group and Capnocytophaga: General Characteristics  Gram-negative bacilli  Require an increased CO2 (5%-10%) environment  Significant cause of endocarditis  Usual flora of the oral cavity  Opportunists in immunocompromised hosts W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  16. 16. HACEK Group H. aphrophilus growing on blood agar Gram stain morphology of H. aphrophilus W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  17. 17. HACEK Group A. actinomycetemcomitans on blood agar. Note the starshaped centers of the colonies. Microscopic morphology of A. actinomycetemcomitans (Gram stain). W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  18. 18. HACEK Group The 48-hour growth of colonies of C. hominis Gram stain of C. hominis showing typical “rosettes” W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  19. 19. HACEK Group E. corrodens on chocolate agar Gram stain morphology of E. corrodens W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  20. 20. HACEK Group and Capnocytophaga Growth of Capnocytophaga colonies on chocolate agar. Compare this growth with Eikenella Gram stain of Capnocytophaga species W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  21. 21. HACEK group Gram stain of Kingella kingae showing plump rods in chains. Compare this morphology with other members of the HACEK group. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  22. 22. Pasteurella species  General characteristics  Colonizes mucous membranes of the upper respiratory tract and gastrointestinal tracts of mammals and birds  Human infections occur from bites and scratches inflicted by animals  Most common isolated species is Pasteurella multocida W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  23. 23. Pasteurella species  P. multocida: Clinical manifestations  Localized infection after a bite or scratch  Respiratory tract infection  Life-threatening systemic diseases (e.g., meningitis, bacteremia) W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  24. 24. Pasteurella Multocida  Culture characteristics  Growth on 5% blood or chocolate shows small, smooth, convex colonies  “Musty” odor  No growth on MacConkey agar; oxidase positive  Microscopic examination  Very small gram-negative rods  Bipolar staining with Giemsa or methylene blue  “Safety-pin” appearance W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  25. 25. Pasteurella Multocida P. multocida growing on blood agar and chocolate agar. No growth on MacConkey agar plate. Gram stain morphology of P. multocida showing bipolar staining W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  26. 26. Case Study  Ten members of a group of 2 dozen retirees from the tobacco industry became acutely ill with pneumonia during a 2-week cruise  The group was staying in a block of rooms together and spent time socializing over drinks and cigarettes in the cabins and saunas and at the poolside W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  27. 27. Case Study  On the fifth day of the cruise, several members went to see the ship’s doctor because of a worsening cough  Chest x-rays revealed patchy lobar pneumonia in all affected individuals  The condition improved with erythromycin therapy W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  28. 28. Legionella Species  General characteristics  Ubiquitous gram-negative rods  Acquired by humans primarily through inhalation of aerosols  Clinical infections  Febrile disease with pneumonia (Legionnaire’s disease) and extrapulmonary involvement  Pontiac fever (without pulmonary involvement)  Asymptomatic infection W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  29. 29. Legionella pneumophila Gram stain of specimen showing intracellular and extracellular Legionella pneumophila L. pneumophila in specimen stained by direct fluorescent antibody technique. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  30. 30. Legionella pneumophila A B (A) Nonselective buffered charcoal yeast extract (BCYE) plate inoculated with sputum specimen. (B) Selective BCYE inoculated with the same specimen but treated before inoculation. Legionella colonies are the smallest visible colonies. W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  31. 31. Legionella pneumophila L. pneumophila colony on BCYE agar after 3, 4, 5, and 7 days of incubation W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  32. 32. Legionella pneumophila: Identification Schema for identification of L. species W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  33. 33. Points to Remember  Clinical manifestations presented by the patient  The types of infections these organisms produce  The risk factors that predispose susceptible individuals W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  34. 34. Points to Remember  What special growth supplements are required for isolation  Where these groups of organisms are usually found  Characteristic features of the organisms for identification and differentiation among closelyrelated species W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.
  35. 35. Thank you For more details please visit www.indiandentalacademy.com W.B. Saunders Company items and derived items copyright © 2001 by W.B. Saunders Company.

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