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CHAPTER 6

MACCONKEY (+), OXIDASE (+) GRAM (-) RODS


2. Agrobacterium radiobacter, Ochrobactrum anthropii, Psychobacter immobilis,
Achromabaaqcter species, Acidovorax species
Description/ 1. Environmental inhabitants
Characteristics 2. Rarely encountered in human infection

Infections are associated with use of intravenous catheters:


Disease/ 1. Endocarditis
Infections 2. Meningitis
*emu* 3. UT

1. Oxidase (+)
Properties 2. Grown on MacConkey
*OGGG* 3. Glucose oxidizers
4. Grow well on BA, CA, BHIB, &THIO

Agrobacterium radiobacter (Rhizobium) Ochrobactrum anthropi

Group CDC Vd-3 CDC Vd

Reaction & Morphology Gram (-) short, pleomorphic rods Gram (-) medium sized straight rods

1. Nitrate reduction (-) 1. Nitrate reduction (v)


Properties 2. esculin hydrolysis (+) 2. esculin hydrolysis (-)
3. Oxidizes glucose, xylose, & mannitol 3. Oxidizes glucose & xylose, mannitol (v)
4. (+) Catalase, urease

Flagella peritrichous peritrichous

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3.Chryseobacterium species, Empedobacter brevis, Sphingobacterium species
1. Former member of Genus Flavobacterium
Description/ 2. Ability to survive in chlorinated water
Characteristics 3. Contaminants in laboratory culture media (yellow colonies)
*FACES* 4. Environmental inhabitants, occasionally encountered in human specimens
5. Survive in moist areas in hospitals

Disease/Infection Nosocomial Infection

1. Oxidase (+)
Properties 2. MacConkey (+) except Sphingobacterium mizutaii
*OMGGY* 3. Grow well on BA, CA NHIB & Thio
4. Glucose oxidizers
5. Yellow pigmented colonies

Chryseobacterium meningosepticum

Description Formerly Flavobacterium meningosepticum

Group CDC group IIa

1. Neonatal meningitis
Disease 2. Septicemia
*ns* (transmitted from birth canal to neonate)

Reaction & Morphology Gram (-) long, thin, straight rods with swollen ends (appear as “II-form”)

Media BA: large, yellow, smooth, circular with entire edge

Properties 1. Capsulated; nonmotile; Indole (+)


*CNI* 2. Gelatinase, DNase, Catalase (+)
*GDC*

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4.Alcaligens species, Comamonas species, Igella species, Bordetella (nonpertussis species),
Psychrobacter species, Rosemonas species, Shewanella species
1. MacConkey (+)
Properties 2. Oxidase(+)
*MONG* 3. Non glucose utilizers
4. Grow on BA, CA, BHIB & Thio
Alcagensis Bordetella Shewanella
Definition/ Environmental organisms; some are found in moist
Characteristics areas of hospitals
Disease/Infection Nosocomial Infections Clinically significant species
Flagella Peritrichous
Property Citrate (+)
xylosoxidans faecalis pertussis parapertussis bronchiseptica putrefaciens
1. Normal flora of Resp.tract of
mammals; 1. Formerly
dogs, cats, rabbits Pseudomonas
Definition Formerly Achromobacter Transient colonizer of 2. Not part of human flora putrefaciens
xylosoxidens Skin & GIT of man (transmitted to man via
2. Found in
droplet spray during
close contact with environment &
mammals foodstuffs
BA: feather-edged, with
Media BA: smooth, glistening, zone of green BA: small, convex, round;
enitre discoloration; fruity 1-2 days incubation
odor (apple/strawberry)
Flagella Peritrichous Polar tufts
1. Nitrate reduction(+) 1. Nirate reduction (v) Mac (-) Mac (+) Mac (+) 1. Citrate (-)
2. Oxidizes xylose; 2. Does not oxidize Oxidase (+) Oxidase (-) Oxidase (+) 2. Only NFB that
Properties glucose oxidation xylose Motility (-) Motility (-) Motility (+) produces H2S
is slow or (-) Nitrate reduction (+) in the butt of
3. Citrate (+) Urease (+) TSI
1. Pneumonia 1. Otitis media
2. Septicemia 2. Septecemia
Disease 3. Meningitis

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5.Vibrio species, Aeromonas species Plesiomonas shigelloides, Chromabacterium violaceum
1. VIBRIOS: Habitat is water, brackish or marine water
Definition Other genera: fresh water
2. Transmitted to humans via ingestion of contaminated water/seafood
3. Exposure of broken skin to contaminated water
1. Oxidase (+) except Vibrio metschnikoviihio
Properties 2. MacConky (+): colonies resembling eneterics
*OMFFG* 3. Fermenters of glucose
4. Facutatively anaerobic
5. Grow well on BA, CA, BHIB & THIO
Vibrio
1. Gram (-) vibrios
Definition 2. Single polar flagellum
3. Halophilic except Vibrio cholearae & mimicus
Property Nitrate reduction (-)
Media 1. BA: medium to large , smooth, opaque with greenish hue;Vibrio fluvialis, mimicus, damsela can be β-hemolytic
2. Mac: NLF except Vibrio vulnifucs which may be LF
Associated 1. Cholera
with 2. Fluvialis
diarrhea 3. Parahaemolyticus
4. Hollissae mimicus
cholerae
Definition sensitive to acid; anaerogenic
1. NLF, ferments glucose, sucrose & mannite
2. String test (+):
Properties a. 0.5% Na deoxycholate
b. Mucus like string

1. Transport media: Cary-Blair TM, Arnies TM


Media for 2. Enrichment medium: Alkaline Peptone Water (6-8hrs)
Isolation 3. Selective media
a. TCBS: thiosulfate ncitrate bile salt sucrose agar
b. TTGA: tellurite taurocholate gelatin agar

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...................................*continuation of page 4............................

V. cholerae Atypical Non-O-1 V. Asiatic


` O-1 V.cholera V.cholerae cholera cholera parahaemolyticus vulnificus
e O-1 O-139
1. Infective
dose: 10^5 1. Gastroeneteritis 1. Able to
organisms involving shellfish and invade the
1. Agglutinate in antiserum 1. Agglnutinat 1. Same Capable of crustaceans bloodstream
against V.cholerae O-1 e in O-1 biochemical epidemic 2. GIT, never 2. Fatal
antiserum properties as O-1 & invade 2. TCBS: olive septicemia
2. Choleragen: 100x but do group; do not pandemic bloodstream green;does not (patient
more potent than LT produce agglutinate in O-1 spread of ferment sucrose suffering
toxin choleragen antiserum cholera; 3. Severe from liver
cholera diarrhea: 10- 3. String test (-) disease)
3. Responsible for Asiatic 2. Choleragenlike toxin 15L/day 3. Capsulated
cholera enterotoxin; 4. Produces 4. Lactose
mild diarrhea 4. “rice-water Kanagawa’s fermenter;s
stools”; hemolysin (heat ucrose (v),
mucus, stable; colonies are β- mostly (-)
epithelial hemolytic on
cells, vibrios Wagatsuma’s agar
Subtypes Ogawa Inaba Hikojima
O factors A,B A,C A,B,C
Biotypes Classical El
(cholera) tor
Tests:
VP at RT - +
Clumping of - +
chicken RBC
Polymyxin B
susceptibility S R
LAB. 1. Stool culture-polyvalent & monovalent serotypng & biotyping
DIAGNOSIS 2. Direct examination: hanging drop of Alkaline Peptone Water after 6-8 hour incubation
-small rods with darting or shooting star motility; then add V. Cholera O-1 antiserum
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5.Vibrio species, Aeromonas species, Plesiomonas shigelloides, Chromabacterium violaceum,
1. VIBRIOS: Habitat is water, brackish or marine water
Definition Other genera: fresh water
(SAME- 2. Transmitted to humans via ingestion of contaminated water/seafood
PAGE5) 3. Exposure of broken skin to contaminated water
1. Oxidase (+) except Vibrio metschnikoviihio
Properties 2. MacConky (+): colonies resembling eneterics
(SAME- 3. Fermenters of glucose
PAGE 5) 4. Facutatively anaerobic
5. Grow well on BA, CA, BHIB & T
Aeromonas Plesiomonass shigelloides Chromabacterium violaceum
1. Aeromonas hydrophila & sobria: GIT 1. Gastroenteritis 1. Transmitted to man by exposure of broke
diseases skin to contaminated water; may lead to
2. Aeromonas caviae: wound infections & speticemia
rarely bacteremia

1. Gram (-) rods; with monotrichous 1. Gram (-) rods with lopotrichous flagella 1. Gram (-) slighltly curved, medium to large rods
flagellum 2. Sensitive to vibriostatic agent O/129 with rounded ends
2. Resistant to vibriostatic agent 0/129 3. Ferments glucose (A only) 2. BA: round, smooth, convex with some strains β-
Cultural & 3. Ferments glucose (A or AG) 4. Decarboxylates ormithine, lysine emolytic; black or very dark purple; exhibit
biochemical 4. Catalase (+) &arginine ammonium cyanide odor
properties
5. BA: large, round raised, opaque, β- 5. Catalase (+) 3. Presumptive ID can be made based on colonial
hemolytic except A. Caviae which is 6. BA: shiny, opaque, smooth, nonhemolytic appearance, oxidase test& Gram stain
nonhemolytic 7. Does not grown on TCBS *if purple pigment of suspected colonies
6. Do not grow on TCBS interferes with oxidase test, the organism
should be grown under anaerobic condition &
retested
4. Oxidase test must be performed on colonies
from media without a fermentable sugar; afalse
(-) may result if surrounding pH is below 5.1

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CHAPTER 7
GRAM (-) BACILLI & COCCOBACILLI
MacCONKEY (-), OXIDASE (+)

1. Sphingomonas paucimobilis & Similar Organisms


1. Sphingomonas paucimobilis (Pseudomonas paucimobilis, CDC group IIK-1
Organisms 2. CDC group IIe
to be 3. CDC group Iii
considered 4. CDC group O-1
5. CDC group O-2

1. Fail to grow on MacConkey agar


Properties 2. Oxidase (+)
*FOOG* 3. Oxidativeky utilize glucose
4. Grow well on routine media (5% sheep BA, CA, BHIB, Thio)
paucimobilis
Definition 1. Exists in environmental niches such as water, including hospital water systems
2. Not part of human flora

1. Media for isolation: 5% sheep BA, CA, BHIB, Thio


2. Incubated at 35 degrees Celsius in CO2 or in ambient air
LAB. 3. Cultue on 5% sheep BA: small, circular, smooth, convex, bright yllow growth pigment
DIAGNOSIS
*MICGS* 4. Gram stain: Gram (-) medium sized straight bacilli
5. Sphigomonas species (including S. paucimobilis & S. parapaucimobilis)
-oxdize xylose & sucrose
-esculin hydrolysis (+)
-nonmotile in motility medium (moitility is present in wet mount (+))
-yellow insoluble pigment (+)

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2.Moraxella & Elongated Neisseria
Genera & species to be 1. Nonliquefaciens 2. Osloensis 3. Lacunata 4. Canis 5. Atlantae 6. Lincolnii . Elongate 8. Weaver
considered *nolcalew* -normal inhabitants of mucosal surfaces
1. Gram (-) small to medium-sized coccobacilli
2. Do not utilize glucose except N. Elongate subsp.glycolytica
3. Oxidase (+)
4. Most strains grow slowly on MacConkey & resemble the NLF Enterobacteriaceae
Properties 5. Grow well on routine media (5% sheep BA, CA) and in the blood culture broths (BHIB, Thio)
6. Catalase (+), except N. Elongate subsp.glycolytica, N.elongata subsp. Nitroreducens
7. Non motile
8. Sensitive to 10-unit penicillin
9. Elongation in the presence of penicillin is a useful criterion for differentiating them from true cocci
10. Opportunistic organisms of low virulence
Moraxella
1. Normal human flora that inhabit the mucous membranes; some species (M. Osloensis), the urogenital tract; may also colonize the
skin
2. Morphologically similar to Acinetobacter except M. Catarrhalis
3. Nonmotile, oxidase (+), catalase (+)
Definition 4. Asaccharolytic
5. Very sensitive to penicillin
6. Grow slowly on Mac except M. Lacunata, M. Lincolnii
7. M. Lacunata historically associated with eye infections but these infections may be caused by other Moraxella species
8. Infections include
9. Bacteremia, endocarditis, septic arthritis & respiratory infections (M. Lacunata, nonliquefaciens, osloensis; most frequently isolated
species
Lacunata (Morax Axenfield Bacillus) nonliquefaciens osioensis
1. Requires serum for growth
2. Able to liquefy serum so depressions are formed on 1. BA; smooth, translucent to 1. BA: smooth,
the surface of Loeffler’s serum agar slants semiopaque; occasionally, colonies translucent to
Culture 3. BA: small colonies that pit the agar (sunken spread & pit the agar semiopaque
colonies) 2. MC: NLF, if growth
4. MC: NG 2. MC: NLF, if growth 3. Can utilize acetate
Direct Detection Methods Gram stain: coccobacillus or medium-sized rod

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3.Eikenella corrodens & similar organisms
1. Eikenella corrodens
Genera & species to be 2. Methylobacterium species
considered 3. Weeksella virosa
*EMWB* 4. Bergeyella zoohelcum
-not associated with human infections but encountered in clinical specimens

1. Gram stain
-E.corrodens: Gram (-) medium length, straight rod
-Methylobacterium: Gram (-) vacuolated, pale staining, short to medium-length bacillus that may resist decolorization
Properties -W.virosa, B. Zoohelcum: Gram (-) short straight rods that may form “II forms” similar to Sphingobacterium
2. Assacharolytic, oxidase (+)
3. do not grow on MC
4. E. Corrodens & Methylobacterium do no grown on either 5% sheep BA or CA; weekshella & Bergeyelia grow slowly on
these media
5. all grow slowly in the broth media used in blood culture; BHIB, Thio

Eikenella Corrodens
Description/ 1. Most commonly encountered
characteristics 2. Found in mized infectious resulting from bites or clenched-fist wounds
3. The “E” is for Eikenella among HACEK group of bacteria to cause subacute bacterial endocarditis

1. Facultatively anaerobic
LAB.IDENTIFICATION 2. BA: colonies are tiny at 24 hours; mature colonies have moist, clear centers surrounded by flat, spreading growth;colonies
may pit or corrode the agar surface;slight yellow pigmentation in older cultures; sharp odor of bleach
3. Broth medium: grows as discrete granules adherent to the sides of the tube
4. Catalae (-), reduces nitrate to nitrite, hydrolyzes both ornithine & lysisne

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4.Pasteurella & similar organism
Genera & species to be considered 1. Multocida 2. Aerogenes 3. Canis 4. Haemolytica 5.. Bettyae 6. Indologenes (Kingella indologenes)
*MACHBIPDS* 7. Pneumotropica 8. Dogmatis 9. Stomatis
1. Small, Gram (-) bacilli
2. Oxidase (+)
PROPERTIES 3. Nonmotile
4. Wil not grown on MacCOnkey
5. Grow well on routine media: BA, CA, BHIB, Thio
Pasteurella Indologenes
1. Normal flora of RT & GIT
Definition 2. Causes hemorrhagic septicaemia, GIT infections, RTI
3. Most human infections are associated with animal bites
multocida Suttonella indolgenes
1. Most commonly encountered in clinical specimens 1. Associated with eye infections
2. Virulence is associated with polysaccharide capsule 2. Can be separted from
DESCRIPTION/ 3. Can be separated from other species based om pasteurrettae by being Nitrate (-)
CHARACTERISTICS a. (+) reactions for ornithine decarboxylase & indole
3. Differes from Kingella species in
b. (-) urease
4. Small Gram (-) coccoid/rod-shaped bacillus being both indole & sucrose (+)
1. Multocida
SUB SPECIES 2. Septica
3. Gallicida
1. Incubated at 35°C in CO2, or ambient air for a minimum of 24hrs.
2. Colonies
GROWTH a. Convex, small & transluecent, may be smooth/rough,
nonhemolytic
b. Brownish discoloration of medium
c. BA: musty/mushroom smell
1. Oxidase (+)
2. Reduce nitrates to nitites
PROPERTIES 3. Catalase (+) except P. Bettyae,
4. Spot indole (+)
5. Glucose fermenter
PRESUMPTIVE I.D 1. Susceptiblilty testing to 2U PCN

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5.Actinobacillus, Kingela, Cardiobacterium, Capnocytophaga & Similar organisms
Description/ 1. Gram (-) bacilli 2. Dysgonic 3. Capnophilic 4. Most species will not grow on MacConkey agar 5. Oxidase (+)
properties 6. Glucose fermenters: serum is added to the basal fermentation medium enhance the growth & detection of fermentation reactions
7. A.actinomycetemcomitans, C. Hominis & Kingella species are A, C & K of HACEK group (cause slowly progressive bacterial endocarditis)
Actinobacillus Kingella Cardiobacterium
1. Normal oropharyngeal flora of humans
Description/ 2. Oxidae (+)
properties 3. Non-motile
4. Ferment glucose
5. Catalase (-)
actinomycetemcomltans kingae denitrificans hominis
1. Normal flora of human URT
1. Normal flora of human oral 1. Important pathogen in children 1. Nonhemolytic 2. Associated w/ endocarditis
mucosa 2. Bacteremia, skin lesions; associated with septic colonies 3. Isolated onky from blood
2. Grows on BA, CA arthritis & osteoarticukar diseases resembling those of 4. Gram stain
Description/ 3. Growth is enhanced by K. Kingae a. Gram (-) pleomorphic rod
properties CO2 with one rounded end &
a. Broth: tiny puffballs/film 2. Grow on Thayer tapered end, giving cells
b. BA: colonies are rough Martin medium (a tear drop appearance)
& sticky surrounded by 5. Culture
a slight greenish tinge 3. Oxidase (+) & a. Small, slightly α-
c. BHIA: center of the produce A from hemolytic, smooth,
mature colony, 4-6 glucose round, glistening &
pinted star-like config. opage; pitting may be
4. Gram stain: short-very short produced
bacilli (singly, in pairs, 6. Similar to S. indolegenes but
chains and tend to exhibit can be distinguished by: it’s
bipolar staining-dots and ability to ferment mannitol &
dashes of Morse code) sorbitol, indole (+), urease (-
), catalase (-), NO3 (-)
1. Growth is enhanced by increased CO2
LAB. 2. On 5% sheep BA: small colonies with clear zone
ID of hemolysis
a. One monotype: “fried egg” appearance
3. Gram stain: Gram (-) shiort plump bacilli with
squared off ends that may form chains

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CHAPTER 8
GRAM (-) BACILLI & COCCOBACILLI
MacCONKEY (-), OXIDASE (V)
1. HAEMOPHILUS
Other species:
*HAPPPS*
1.haemolyticus
H. influenza H. ducreyi 2.aphrophilus
(Influenza Bacillus Pfeiffer) 3.parahaemolyticus
4.parainfluenzae
5.paraphrophilus
6.segnis

1. Normal RT & occasionally the Genital Tract


2. URTI, suppurative infections & systemic infections (meningitis)

TYPEABLE NON-TYPEABLE H.influenza H.influenza


STRAINS STRAINS (encapsulated) (noncapsulated)

Capsule: Do not produce Etiologic agent of: 1. ACUTE Chancroid/soft Normal flora of
Sugar-alcohol Capsule *ACEPS* CONJUNCTIVITIS chancre humans only
phosphate (polyribitol 1. Acute Bacterial Meningitis (pink eye- biogroup (veneral
phosphate-PRP) 2. Cellulitis w/ bacteremia disease)
aegypticus)
3. Epiglottis *brazillian purpuric fever
4. Pneumonia 2. ACUTE & CHRONIC
5. Septic arthritis 78 OTITIS MEDIA
6 groups
A,B,C,D,E,F Normal inhabitants of
the URT
*type B: (upper respiratory tract)
serious infections in
humans (systemic)
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2. SPECIMENS
*BCMJRS* must be plated
1. Blood IMMEDIATELY H. ducreyi:
2. CSF
3. Middle ear exudates
4. Joint fluids 1. Cotton swab moistened 2. Swabs are placed in 3. Gram stain: Gram (-)
5. RT specimens with PBS to collect modified Stuart’s TM or coccobacilli resembling
6. Swabs from inflamed material from ulcer base Arnie’s charcoal TM “SCHOOL OF FISH”
Conjunctivae &
Abscess drainage

3. MEDIA ISOLATION
CA supplies factors necessary for growth
of most Haemophilus species: 1. Horse BA, rabbit BA, sheep blood- NADase
HEMIN= X factor NAD= V factor 2. Haemolytic staphylococci, pneumococci, neisseriae
(HIDA) 3. Satellite phenomenon
1.H. Haemolyticus All clinically 4. Horse blood bacitracin: selective medium to increase isolation
2.H. Influenzae significant species a. H. Ducreyi: Mueller Hinton base CA with 1% IsoVitaleX & 3micrograms/ml vancomycin
3.H. Ducreyi except H. 5. Heart infusion base agar 10% fetal bovine serum + 3 micrograms/ml vancomycin
4.H. Aphophilus Aphropilus & H.
(for initial isolation) Ducreyi require V
factor
.

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4. CHARACTERISTICS
FACULTATIVE ANAEROBES Growth stimulated by 5-10% CO2 35-37 °C Grow within 24 hours
in a humidified atmosphere (H. Duceyi for 7 days)

5. LABORATORY IDENTIFICATION
1. CA: Transparent, Moist, Smooth, & Convex colonies (dew drop) with mousy/bleachlike odor
2. Gram stain: Gram (-) pleomorphic coccobacilli

1.TEST FOR X & V FACTOR 2.BIOCHEMICAL 3.SEROLOGY


REQUIREMENT TESTS

1. PORPHYRIN TEST 1. Rapid ID of Hib-polysaccharide capsule (PRP)


-α-aminolevulinic acid-
porphyrins or protoporphyrins ALL ARE OXIDASE (+)
except H. segnis
2. Use of filter paper strips or 2. Used directly on clinical specimens like CSF, serum, & urine
discs impregnated with X and V
factors

3. Satellite phenomenon 3. Same tests can be performed on isolates: *CQLCERF”


a.Coagglutnation test
b.Quellung Reaction
c.Latex aggluytination test
d.CIE, EIA, RIA, FAS

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