MD-2021 | Manila Central University S.Y.

2018-2019

MICROBIOLOGY
NOCARDIA, ACTINOMYCES, Haemophilus influenza
HAEMOPHILUS and BORDETELLA - Coccobacillary
Based on the lecture of Dr. Gironella | January 10, 2019 - Fastidious
- Hemo (blood), philus (loving)
- Obligate parasite of humans and animals
- Exhibits pronounced host specificity
Actinomyces israelii
H. influenzae
- Gram (+) bacilli short or long, thin filaments or club-shape , H. aegypticus (Koch-Weeks Bacillus)
slow growers H. parainfluenza
- some aerotolerant or anaerobes (facultative) H. ducreyi

- chronic, suppurative, granulomatous infection Cultural Requirements

- characterized by pyogenic lesions with interconnecting sinus
(tract with only one opening, compared to fistula which has
two) tracts containing granules of microcolonies of bacteria
embedded in tissue elements
o SULFUR GRAINS or GRANULES
3 Forms of Infections
1. Cervicofacial “lumpy jaw”
2. Thoracic or Pulmonary infection: fever, cough, purulent
sputum, sinus tract to chest wall
3. Abdominal, General Surgery, Women with intrauterine
devices: ruptures appendicitis or ulcer
Clinical Infection
- Male predilection, 15 to 35 y/0
- Chocolate agar is the most commonly used
- Levinthal and Fildes agar may also be used (Levinthal is
useful in differentiating between encapsulated and
noncapsulated strains)
o Encapsulated – more invasive infections
- Requires 10% CO2
Nutritional Requirements
- X factor (protoporphyrin IX, precursor of hemin)
- Coenzyme I or V factor [nicotinamide adenine dinucleotide
(NAD)]
- Exhibit phenomenon of “satellitism”
o Haemophilus is cultured with Staphylococcus
which produces NAD (V factor)
o

Pathogenesis
- TRAUMA → mucus and epithelial surfaces
- ASPIRATION → pulmonary/thoracic infections where
organism grows anaerobically in deeper tissues → cellular
infiltrations → SINUS TRACT FORMATION

Diagnosis
- Sputum, pus collection → sulfur granules
- Culture (thiogycolate broth as the transport medium) → BHIA
“molar tooth colonies”

Treatment
- Penicillin is DoC (6 to 12 months)
- Surgical excision/drainage
- Alternative drugs: tetracycline, erythromycin, clindamycin or
sulfonamides
X FACTOR V FACTOR
H. influenzae + +
H. parainfluenzae - +
Nocardia asteroides H. ducreyi + -
H. haemolyticus + +
- Found worldwide in soul and H20
- Aerobic, gram (+) and partially acid-fast bacilli (has shorter-
chained mycolic acid) BIOTYPES
o Other organism with mycolic acid are: ➢ Biovar I – meningitis
Mycobacterium and Corynebacterium o Urease, ornithine decarboxylase and indole
- Catalase (+), urease (+) positive
➢ Biovar II and III – conjunctivitis, chronic bronchitis, sinusitis
Pathogenesis and otitis media
- Risk factors with immunocompromised pxs, corticosteroid tx,
organ transplant, TB, alcoholism Antigenic Structure
- Fever, weight loss, chest pain - Major classes of surface antigens:
- Mimic TB, lobar pneumonia, abscess formation o Capsular polysaccharide (virulence)
Diagnosis o Lipopolysaccharide
- Gram (+), AFB (+) o Outer membrane proteins
- No serologic test - Capsular antigens
Treatment o Major antigenic determinanat
- DoC: Trimethoprim- Sulfamethoxazole o Antibodies are age-dependent

©icacomedian 1
 MD-2021 | Manila Central University S.Y. 2018-2019

o Confers type specificity - Otitis media

o Basis of grouping into serovars a, b, c, d, e, f o Ampicillin or amoxicillin – DoC
▪ Most invasive is serotype b o Alternatives:
• Polyribose, ribitol (pentose) ▪ Co-trimoxazole
• Hib vaccine ▪ Penicillin (or erythromycin) +
▪ A, b, c and f – teichoic acid type sulfonamide
▪ Cefaclor
TYPE SUGAR
a Glucose, techoic acid ***observe patient for 3 days to know if the patient is resistant with the
b Polyribose, ribitol (pentose) drug (usually presents with fever, seizure, deteriorating level of
C Galactose, IgG – B and O consciousness, etc.)
D Hexose, IgM – B
***infants with meningitis whose fontanelles are still open must be
E Hexosamine polysaccharide IgA – X
monitored with frequent measurement of head circumference
F galactosamine
-base of the brain (scarring) → block CSF → hydrocephalus
Case reactions with E. coli K-Ag
Active Immunization
o Serovar b polymer is unique
- PRP-T-Tetanus toxoid conjugate: <5 y/0
▪ Stimulates antibodies
▪ Responsible for invasiveness
Passive Immunization
▪ Cross-reaction with other bacteria with
- High risk group:
capsule
o Sickle cell anemia
o Asplenia
Determinant of Pathogenicity
Rifampicin Prophylaxis
- Phosphoribosylribitol phosphate (PPP) capsule
- Children exposed to household or day care
- Outermembrane proteins and LOS
o Attachment, invasive, and resistance to
phagocytosis
o LOS (endotoxin) paralyzes ciliated respiratory
epithelium
Haemophilus aegypticus
*H. influenzae – most common cause of meningitis in 6months to 6y/o
- Indistinguishable from H. influenzae biovar III
Epidemiology o Referred to as the H. influenzae biotype
- Unencapsulated organisms are carried in the nasopharynx aegypticus
of symptom-free persons - Causes purulent conjunctivitis (“pink-eye”)
- Rates of carriage is 60%-90% in healthy young children and o Responds to topical sulfonamides
35% in adults - May cause severe invasive disease similar to
- Frequency of invasive infection is inversely proportional to meningococcemia caused by N. meningitidis
age

Pathogenesis Haemophilus parahaemolyticus

- Invades the respiratory tree → go to the blood → cross BBB
as it resists phagocytosis → MENINGITIS - Normal flora of mouth and nasopharynx
- Predisposing factors for disease:
Clinical Manifestations o Dental disease and procedures
- Meningitis – most common o Dental procedure and oral trauma
o Rare before 3 months and uncommon after 6 o Respiratory tract infection
years of age o Alcoholism
o Children must receive full dose of Hib - Most common infection – endocarditis (5%)
- Epilogttitis - Tx: Ampicillin + _______
- Cellulitis – children <2 y/0 (cheek is commonly involved)
- Bacteremia without local disease

Other Infections
- Pneumonia, otitis media, septic arthritis
Haemophilus ducreyi
-
Diagnosis - Causes chancroid – STD
- Direct exam (GS) [compared with chancre (not painful) – “syphilis”]
- Culture and isolation: chocolate agar, IsoVitaleX o Single or multiple sharply circumscribed,
- Antigen detection: nonindurated PAINFUL ulcers usually in inguinal
o Countercurrent immunoelectrophoresis (CIE): area
rapid o “school of red fish” appearance in microscopy
o Latex particle agglutination o CAP with vancomycin
o ELISA - Tx: erythromycin, cotrimozaxole

Treatment
- Meningitis and epiglottitis
o Ampicillin + Chloramphenicol
▪ Highly resistant strains: Ceftriazone

©icacomedian 2
 MD-2021 | Manila Central University S.Y. 2018-2019

3 STAGES OF DISEASES
Bordetella pertussis 1. Catarrhal or Prodromal Period – 1 to 2 weeks, mild S/S of URTI

MORPHOLOGY 2. Paroxysmal Period - 1 to 6 weeks, paroxysmal cough

- Gram-negativce coccobacilli, singly, in pairs, and in small - “Whooping cough” – inspiratory breath takes place thru
clusters, bipolar metachromatic granules narrowed glottis
- Strict aerobes - Increased WBC (12000 to 20000)
- Primary cultures – uniform in size but in subculture - Excessive mucus and vomiting
pleomorphic, filamentous and thick bacillary forms are
common 3. Convalescence period – 2 weeks of coughing
- Non-motile except bronchoseptica *** 4 to 8 weeks duration of illness from catarrhal to
convalescence period
EPIDEMIOLOGY ***diagnosis in catarrhal stage will cut the duration to only 4
- Highly communicable weeks of cough
- Multoplu in cilia of epithelial cells (tropism)
Complications:
CULTURAL GROWTHS - CNS: neurologic sequelae due to anoxia
- Modified Bordet-Gengou (Potata Glycerol in Blood) - Insterstitial pneumonia – main cause of mortality
o Primary isolation - Sinusistis, otitis media
PHASE: - Emphysema/atelectasis
I – smooth (virulent) - Flare-up of PTB (give Isoniazid for prevention)
II and II – moist (intermediate
IV – rough (less virulent) ***TRIPLE DRUG: macrolide (for pertussis), acetaminophen (for
affinity to Strep and Haemophilus) and Isoniazid (for prevention of
Antigenic Structure PTB)
- Surface O Ag – found in all species
Diagnosis
AGGLUTINOGENS 1. DEFINITIVE – isolation of B. pertussis (best time at catarrhal phase)
- Based on heat labile 120’C or capsular Ag of Kauffman
- Serotype numbered - nasopharyngeal swab, cough plate,
o Factors 1 to 6 – B. pertussis - Mod. Stuart’s medium, Mishulow’s charcoal medium
- Bordet-Gengou – recommended for primary isolation
Determinants of Pathogenicity o “scattered pearl” appearance
1. PERTUSSIS TOXIN - Regan-Lowe Medium
a. Histamine sensitizing factor (HSF)
▪ Thermostable; with subunits 2. SEROLOGIC - Immunofluorescence
▪ MW 73, 000 to 77, 000
b. Lymphocytosis promoting factor (LPF) Tx:
▪ Adherence to post-capillary venules - Erythromycin – DoC
▪ Responsible for relative lymphocytosis o Shorten the clinical course
▪ Polyclonal activator of human T- - Tetracycline
lymphocytes - Trimeth-sulfa
c. Islet-activating protein (IAP) - Chloramphenicol
- Supportive treatment: HYDRATION!

2. HEMAGGLUTININS – mediate adherence to mammalian

respiratory cilia
a. Fimbriae-HA (F-HA) – attract antibodies, attaches
cholesterol
b. Pertussis-HA (P-HA) – attaches to sialic acid

F-HA and PT-HA increases adherence of H. influenzae and -END-

S. pneumonia in mixed infections

3. CHROMOSOMAL LOCUS in B. pertussis

- bvgA - related to activation of virulence genes (adenylate
cyclase)
bvgS – environmental factors

4. Heat Labite Toxin

5. LPS – Heat stable toxin
6. Tracheal cytotoxin – ciliostasis (stops movement of the cilia)

Epidemiology
- Number of cases – immunization
- Highly communicable
- Carriers are immune

©icacomedian 3