ILIGAN MEDICAL CENTER COLLEGE

SEMINAR 1: SPECIAL TOPICS

BY: Samantha R. Namoc, RMT
PARVOBACTERIA
GRAM (-)COCCOBACILLUS, FASTIDIOUS
AEROBIC, MAC -, NEEDS CO2
A. Haemophilus
Require X (hemin) and V (NAD); Satellitism (H. influenza)
Medium: CAP (Horse blood) + 5%CO2; No growth on MacConkey agar
*Nitrate reducer
Haemophilus
*ferment CHO
*Non motile
*Catalase+
*CAP+ ; BAP-

X and V factor

XV+ V+ X+ XV-

*H. influenza *H. parainfluenza H. ducreyi H. aprophilus

*H. aegyptius *H. paraaprophilus
*H. haemolyticus *H. parahaemolyticus

1. H. influenzae (Pfeiffers Bacillus) GNCB

Virulence: capsule, IgA protease, LPS, pili
Satellite around S. aureus (BAP); #3 cause of bacterial meningitis
Major cause of acute epiglottitis; Cystic fibrosis (Horse blood bacitracin agar)
Beta-lactamasea (+) control = cefinase disc test
Lab diagnosis
a. Culture on CAP, Levinthal, Fildes
Grayish, dew drop colony, mousy
b. X and V factor
c. Satellitism (BAP + S. aureus)
d. Porphyrin test = negative

2. H. ducreyi (chanchroid)
Soft chancre; school of fish
Growth on CAP + Vancomycin
3. H. aegyptius
Koch weeks bacillus
Pink eye conjunctivitis; Brazilian purpuric fever
A. Bordetella pertussis
Capsule, obligata aerobe
Whooping cough bacillus
Catarrhal, paroxysmal, convalescence
Nasopharyngeal swab (carrier)
Require cysteine & methionine for growth
MAC + except B. pertussis
Culture media:
a. Mercury drop (pearl like) colonies on Potato blood glycerol agar or Bordet Gengou
b. Regan Lowe (charcoal horse blood)
c. Jones Kendrich (charcoal, yeast extract)
d. Charcoal cephalexin blood agar (CCBA)
e. Steiner and Scholte, Casamino broth
Species MOTILITY UREASE OXIDASE MAC&BAP
B. petussis - - + -
B.parapertussis - + - +
B.bronchiseptica + + + +
C. Brucella spp.
No capsule, obligate aerobe, non motile
Zoonotic Erythritol enhance growth
Tryticase soy broth(TSB), Wisconsin (W), Castaneda broth(biphasic medium)
Brucellosis, uundulant fever, malta fever
Bangs dis, animal abortion, andocarditis, Cyprus fever, laboratory acquired infection, mediterranian fever
SPECIES UREASE CO2 THIONINE FUCHSIN
B.abortus + + - +
(bangs dis) HS2
B.melitensis + - + +
B.suis H2S + - + -
B.canis + - + -

D. Francisella tularensis Cystine & cysteine require

Urease (-), Mac (-), non motile, capsule, aerobe
Tularemia Ohara, market mens disease, Deerfly, lemming. Rabbit fever, watery-trappers; laboratory acquired infection
Catalase +, oxidase -, beta-lactamase +
Culture media:
a. GCBA GLUCOSE CYSTEINE BLOOD
b. PCA PEPTONE CYSTEINE AGAR
c. CHA CYSTEINE HEART AGAR
E. Pasturella multocida GNCB
Capsule, non motile, bipolar stain safety pin
Oxidase +, catalase +, glucose+, ornithine +, indole+, urease+
Growth on BAP but not in MacConkey
 Animal bite wound, Pneumonia, endocarditis, meningitis, arthritis

HACEK : SBE, CO2, (+) Blood, (-)MAC, GNCB

H = Haemophilus influenza
A = Actinobacillus actonomycetemcomitans
C = Cardiobacterium hominis
E = Eikenella corrodens
K = Kingella kingae
OXIDASE CATALASE FEATURES
H +/- - NO X and V

A - + STAR LIKE COLONY

C + - INDOLE (+)

E + - ASSACHAROLYTIC

K + - TWITCHING MOTILITY

SPIROCHETES

1. Treponema pallidum
Non cultivable on agar medium
Obligate intracellular (rabbits testicle) SPECIES DX DISEASE
Syphilis
a. Primary hard chancre
Treponema SEROLOGY Syphilis, yaws, pinta,
b. Secondary condylomata lata, skin rash
bejel
c. Latent asymptomatic (serology)
d. Tertiary gummas Leptospira CULTURE Weils / infectious
e. Congenital syphilis stillbirth, abortion DAP cell wall jaundice
f. Tx: Penicillin (herxheimer reaction)
Other Treponemes
a. T. pertenue yaws/framboise
b. T. carateum pinta Borrelia GIEMSA, SEROLOGY Lyme, relapsing
c. T. endemicum bejel (blood/BM) fever
Lab diagnosis
1. Dark field microscopy corkscrew motility
2. Levaditi silver impregnation
3. Serology
a. VDRL, RPR, TRUST (regain test)
b. FTA-ABS, TPHA, MHA-TP, HATTS (Trep. Ab test)

2. L. Interrogans icterohemorrhagica
Spiral with hook ends, weil disease (animal urine)
1st week blood, CSF/tissue (acute)
2nd week urine (chornic)
Culture media:
a. Fletchers = animal serum + fatty acid (30C, 6 wks)
b. MAT MACROSCOPIC AGGLUTINATION TEST = screening test
SERUM +Ag (killed leptospira); (+) AGGLUTINATION
c. MIT MICROSCOPIC AGGLUTINATION TEST = confirmatory
SERUM +Ag (LIVE LEPTOSPIRA); (+) AGLUTINATION (DARKFEILD)

3. Borrelia (Blood Spirochete)

A. Relapsing fever due to
Borrelia recurrentis (epidemic) louse
Borrelia anserine, toricatae, parkeri (endemic) tick
DX: Wright / Geimsa blood / Bone marrow
B. Lyme disease (B. burgdorferi)
Tick bite (Ixodex dammini)
1st stage erythema chrinocum migrans
Culture on Barber Stoenner Kelly (BSK) at 33C for 6 weeks
Chlamydia
Obligate intracellular parasite
Energy ATP parasite
Inclusion body (Geimsa stain) - Diagnostic
Elementary body infectious
Reticulate body reproductive

1. Chlamydia trachomatis = TRIC agent (trachoma, inclusion conjunctivitis)

LGV lymphoma granuloma veneruem(BUBOES) STD (Frei); #1NGU(non gonococcal urethritis), PID; Sensitive to sulfonamide
Reiters syndrome
Lab diagnosis TRANSPORT = 4C
a. Glycogen Inclusion body (halberstadter prowazeik) iodine/geimsa
b. McCoy (BEST MEDIUM) GOLD STD.
c. Direct Fluorescence Assay Chlamydial Ag
d. PCR/NAAT = BEST METHOD

2. Chlamydia psittaci
Parrot fever or psittacosis (ornithosis) disease of birds
Man: pneumonia
Non glycogen inclusion body (Geimsa) Levinthal Cole Lillie
Resistant to sulfonamide

3. Chlamydia pneumoniae (TWAR TAIWAN ACUTE RESPIRATORY)

Human to human (pneumonia); growth on Human lines & Hep-2 cell
Immunofluorescence test, PCR
 Rickettsia
Obligate intracellular except Coxiella, Arthropod borne
Cross react with Proteus (weil felix)
Lab diagnosis
Stain = Gimenez, Macchiavelo, Castaneda, Giemsa
Culture = Embryonated egg (best), cell culture
Weil Felix test Rickettsial Ab
Rickettsial spp Vector
R. rickettsi Tick Rocky Mountain Spotted Fever
R. akari Mite Rickettsial pox
R. typhi Rat flea Murine (endemic) typhus
R. prowazekii Louse Epidemic typhus & Brill-Zinsser dse
Orientia tsutsugamuchi Chigger Scrub typhus
Bartonella Quintana louse Trench fever
Ehrlichia chaffensis Tick Monocytic Ehrlichoisis
Ehrlichia equi (morulae) Tick Granulocytic Ehrlichoisis
Coxiella burnetti (excluded) Q fever (inhalation/tick)
Mycoplasma
wall less (pleomorphic)
fried egg / mulberry colony (Dienes stain)
require STEROL for growth except acholeplasma

1. M. pneumoniae (Eaton agent)

Pleuropneumoia like organism (PPLO)
PAP or walking pneumonia
Fried egg colony (aerobic w/ CO2)
Selective: PPLO agar, Edward Hayflicks
Confirm: Hemadsorption test
Best: inhibition of growth by specific antisera

2. M. hominis
Large fried egg colony, Post abortal, Post partum fever
Media: A7/A8, NYCA

3. Ureaplasma urealyticum
T strain (tiny fried egg), urease +, NGU
Media; A7/A8, NYCA
MISCELLANEOUS BACTERIA
1. Gardnerella vaginalis
Oxidase and catalase NEGATIVE
Cause BACTERIAL VAGINOSIS, CLUE CELLS on cytologic exam = (GS, PAP, WET MOUNT)
FISHY AMINE like odor (alk.ph whiff/sniff test) rgt: 10% KOH
Media: Human blood tween 80 agar(beta. Hem), Columbia agar, SPS sensitive
2. Calymmabacterium (Klebsiella) granulomatis
Safety pin, capsule, Donovan bodies (Giemsa)
Granuloma inguinale (donovanosis)
3. Actinobacillus actinomycetemcomitans
Start like colony, SBE
Dots and dashes of Morse code
4. Streptobacillus monoliformis
String of beads, fluff balls (broth)
Fried egg, SPS sensitive, rat bite fever, Haverhill dse (ingeston)
Note: spirillium minus/minor rat bite fever, spiral
5. Chromobacterium violacuem
Violet colored, Violacein; NH4 cyanide, MAC-NLF
6. Cardiobacterium hominis
Cause SBE, Tear drop shape
7. Capnoctophaga spp.
Gliding motility (spreading col), Periodontal dse. (oral flora), Fusiform rod.
NO3+, Esculin +
Oxidase and catalase NEGATIVE
8. Bartonella henselae
Cat scratch, Bacillary angiomatosis, Peliosis hepatitis
9. Bartonella bacilliformis
Destroy RBCs
Vector: SANDFLY
Carrions disease
Verruga peruana skin eruption and Oroya fever anemia
10. Legionella pneumophila = shirt gram (-), aerobic
Require L-cystine and iron for growth
Air conditioning; water cooling
broadstreet pneumonia & Pontiac fever LISTERIA ERYSIPELOTHRIX
Transport = 4C ; storage = -70C Catalase + -
Lab diagnosis
a. DFA Legionella Ag Motile(25c) + -
b. Culture best on BCYE = blue green colony / cut glass colony
Hemolysis beta Alpha
c. Stain: dieterle silver stain (black)
VP + -
11. Listeria monocytogenes
Gram (+) rod, Motile at RT ; TUMBLING mot. (broth); umbrella like H2S - +
motility
BE & HIPP + -
Beta hemolysis on SBAP; Cold enrichment at 4C (+) & McBride (+)
Listerolysin O O2 labile hemolysis; Anton test Ocular virulence test Gluconate + -
Meningitis, sepsis, Infantiseptica granulomatous, food poisoning
12. Erysipelothrix rhusiopathiae Media McBride, Cold BAP
Gram (+) rod, non motile, H2S prod, Catalase (-) enrichment
Test tube brush, erysipiloid (Butchers cut)