Pathologic and Non-Pathologic Bacteria  Some strains of P.

vulgaris may ferment sucrose

Notes to Remember:
> K. pneumoniae subsp. ozanae is the causative agent
of chronic atrophic rhinitis ("foul smelling" atrophic
rhinitis).
> K. pneumoniae subsp. rhicoscleromatis causes a
granuloma of the nose and oropharynx.
> K.oxytoca and K. ornithinolytica are both indole
positive.
and give an acid/acid reaction in TSI agar. PAD
test: positive
 LIA reaction: R/A
 IMVIC reaction: -+vv (P. mirabilis); +++v (P.
vulgaris)
 TSI reaction: K/A, (+) gas, (+) H2S -P. mirabilis
 K/A, (+/-) gas, (+) H2S - P. vulgaris

ENTEROBACTER PROVIDENCIA
 It resembles Klebsiella when growing on Mac  Is one of the causes of nosocomial outbreaks
Conkey agar. involving burn units.
 Culture: pink colonies and maybe with mucoid  Species: P. alcalifaciens, P. stuartii, P. rettgeri, P.
colonies - Mac Conkey agar rustigianii and P. heimbachae Urease test:
 Significant species: E. aerogenes, E. cloacae positive (P. rettgeri)
(common isolate), E. gergoviae and E.  PAD test: positive
hormaechei  IMVIC reaction: ++++
 TSI reaction: K/A, (-) gas, (-) H2S
PROTEUS
 It is isolated from urine, wound and ear PROVIDENCIA RETTGERI
infections.  Is a pathogen of the urinary tract.
 It can infect the proximal kidney tubules and can  It also causes diarrheal disease among travelers.
cause AGN, particularly in patients with urinary
tract defects and catheterization. PROVIDENCIA STUARTII
 It is a rapid urease producer - urease splits urea  It has been isolated from nosocomial outbreaks
in urine, raises urine pH and encourages renal in burn units and in urine cultures.
stone formation.  It is mostly resistant to antimicrobial agents
 Species: P. mirabilis (common isolate), P. (together with P. rettgeri).
vulgaris, P. penneri, P. myxofaciens Human
pathogens: P. mirabilis and P. vulgaris PROVIDENCIA ALCALIFACIENS
 Culture: "swarming phenomenon" and "burnt  Is most commonly found in the feces of children
chocolate" or "burnt-gun powder" odor The with diarrhea.
swarming phenomenon is a result of a tightly
regulated cycle of differentiation from standard MORGANELLA
vegetative cells (swimmers) to hyperflagellated,  Same biochemical reaction with P. vulgaris
elongated, polyploidy cells (swarmers) capable except citrate negative.
of coordinated surface movement.  Species: M. morganii
 PAD test: positive
 IMVIC reaction: ++--
 TSI reaction: K/A (+) gas, (-) H2S
 Other Biochemical Tests: Positive in Urease,
KCN, Ornithine Decarboxylase
CITROBACTER FREUNDII
 It can be isolated in diarrheal stool cultures
(extraintestinal pathogen).
 It has been associated with endocarditis in IV
drug users.
 The colony morphology on primary plated media
can be mistaken for that of Salmonella when
isolated from stool cultures, since majority of the
strains produce H2S and a few fail to ferment
lactose.
SALMONELLA
 The most serious pathogenic enterobacteria for
humans, causing enteric fever (typhoid fever)
and acute gastroenteritis (food poisoning).
 It inhabits the GI tract of animals.
 Humans acquire this organism by ingestion of
contaminated animal food products or
improperly cooked poultry, milk, eggs and dairy
products.
 It may also be transmitted by human carriers.
 Culture: MAC - clear, colorless colonies
Media with H2S indicators - colonies with
black centers (HE, BSA and XLD)
SSA - colorless colonies with black centers
 Species: S. enterica (type species) and S. bongori
 Subspecies of S. enterica: S. enterica subsp.
enterica (1), S. enterica subsp. salamae (II), : S.
enterica subsp. arizonae (Illa), S. enterica subsp.
diarizonae (Illb), S. enterica subsp. houtenae (IV)
and S. enterica subsp. indica (VI)
 Virulence factors: fimbriae and enterotoxin
 Antigenic structures: somatic O and flagellar H
(for serologic grouping) Vi antigen (S. serotype
Typhi)- antiphagocytic
 Main etiologic agent of enteric fever: S. serotype
Typhi
 Etiologic agents of paratyphoid fever: S. serotype
Paratyphi A, B, C, and S. serotype Choleraesuis
3 General Categories of Salmonella Infection:
1. Gastroenteritis
 Is one of the most common forms of "food
poisoning".
 The salmonella strains associated with this
infection are those found in animals, mostly S.
enterica subsp. enterica.
 For the peanut-butter outbreak, S. serotype
Typhimurium is the causative agent.
 The used of contaminated cooking utensils and
cutting boards can spread the bacteria to other
food.
⚫ inadequate refrigeration also allows the growth and
multiplication of the organisms.
 Sources of infection: poultry, eggs and egg
products, milk, and handling of pets
 Infective dose: 106 bacteria
Symptoms: nausea, vomiting, fever and chills, watery
diarrhea and abdominal pain.
2. Enteric Fever
 is also known as typhoid fever, and it is caused
by S. serotype Typhi
 It is a febrile disease that results from the
ingestion of contaminated food originating fro
infected individuals or carriers.
 Direct transmission through fomites is also
possible.
 Sources of infection: human carriers,
contaminated food and water
 Causes of outbreaks: improper disposal of
sewage, poor sanitation and lack of modern
water system
 Symptoms: malaise, anorexia, lethargy, myalgia
and continous frontal dull headache The
characteristic "rose spots" appear during the 2nd
week of fever.
 The gall bladder is the site of long-term carriage
of S. serotype Typhi.
 Complications: necrosis in the gall bladder
(necrotizing cholecystitis) and Peyer's patches
 S. serotype Typhi is resistant to gastric acids and,
on reaching the proximal end of th small
intestine, it invades and penetrates the intestinal
mucosa
3. Bacteremia
 It occurs with and without extraintestinal foci of
infection caused by nontyphoidall Salmonella (S.
Typhimurium, S. Paratyphi and S. Cholerasuis).
 It is characterized by prolonged fever and
intermittent bacteremia.
Notes to Remember:
 "Rose spots" (blanching rose-colored papules)
appear around the periumbilical region, and it is
a sign of infection.
 Individuals who recover from the infection may
harbor the organisms in the gall bladder, which
becomes the site of chronic carriage.
 Carriers of Salmonella excrete the organisms in
their feces continuously or intermittently. The
carrier state may be terminated by antimicrobial
therapy if gallbladder infection is not evident.
 Cholecystectomy is the only remedy to the
chronic state of enteric carriers.
Specimens for Salmonella Identification:
 1. Blood 1" week of infection
 2. Stool-2 week of infection
 3. Urine-3 week of infection
SHIGELLA
 It is closely related to the genus Escherichia
 It not a member of the normal gastrointestinal
flora.
 It is an intracellular organism - they multiply
within the cells of the colon epithelium.
 It is transmitted by flies, fingers, food and feces
(the four F's) and water by infected persons
(fecal-oral route).
 Reservoir: humans only; no animal reservoir
 Culture: MAC- clear, fragile, NLF colonies
SSA-colorless colonies without black centers
 Virulence factor: Shiga toxin
 Species: S. dysenteriae (most virulent), S.
flexneri ("gay bowel syndrome),
S. boydii and S. sonnei
 Antigenic structures: somatic O
 Specimen: rectal swab
 Several serotypes exist within each species,
except S. sonnel, which has only one serotype.
 Clinical Infections:
Plague
 Is a disease of the rodents transmitted to humans
by fleas.
 It is carried by urban and domestic rats and wild
rodents.
 Humans may also be infected by ingestion of
contaminated animal tissues and inhalation
 of contaminated airborne droplets.
 Once inside the human body, the bacteria
multiply in the blood and lymph.
2 Forms of Plague:
1. Bubonic plague
It is associated with high fever and painful
inflammatory swelling of axilla and groin (buboes).
It results from the bite of an infected flea.
2. Pulmonary plague
It is acquired by close contact with other victims.
It occurs secondary to the bubonic plague.
Yersinia enterocolitica
 It is the most commonly isolated species of
Yersinia.
 It is the causative agent of enterocolitis-
waterborne gastroenteritis.
 It is motile at 22°C but not at 35°C; with V and
W antigens; ferments sucrose and mannitol.
 ⚫It requires cold enrichment technique (4°C)
using phosphate buffered saline for several
weeks for isolation from fecal material.
 It has the ability to survive in cold temperatures
(food refrigeration).
 It has been isolated also from contaminated
packed RBCs (blood transfusion).
 It grows on routine culture media (BAP and Mac
Conkey); grows best at 25"-30°C.
 Mode of acquisition: Consumption of
incompletely cooked food (pork and pork
intestines, and vacuum-packed meat) and dairy
products (chocolate milk), and handling pets
 Microscopy: cocccobacilli with bipolar staining
 Culture: CIN- "bull's eye colonies" (dark red or
burgundy centers with transparent borders) at 48
hours
 Selective medium: Cefsulodin-irgasan-
novobiocin (CIN) agar
 Reservoirs: swine, dogs, cats, rabbit and cattle
 Other related infections: appendicitis-like
syndrome, arthritis, and erythema
 IMVIC reaction: v+-- nodosum
 TSI reaction: K/A, (-) gas, (-) HS
GRAM-POSITIVE ANAEROBIC
SPOREFORMING BACILLI
 They are CHO fermenters except C. tetani and C.
histolyticum.
 Some members are killed immediately (C. novyi)
in the presence of oxygen.
Clostridium perfringens (Gas gangrene bacillus)
 It is formerly known as Clostridium welchii.
 It is the most commonly isolated member of
Clostridia in blood cultures.
 It produces deoxyribonuclease-lowers viscosity
of exudates; for mobility of bacteria. It lacks the
ability to produce a number of essential amino
acids
 It causes outbreak after ingestion of
contaminated meat and gravy.
 Virulence factors: a-toxin and enterotoxin
 Microscopy: "boxcar"-shaped bacilli
 spores are seldom seen, but it is oval, central to
subterminal
Culture: BAP-dome-shaped, gray to white colonies
BAP double zone of hemolysis (alpha and
beta zones) Litmus milk - "stormy fermentation of
milk"

CLOSTRIDIA
 Is an obligate anaerobic, Gram-positive
sporeforming rod; catalase (-).
 Is frequently encountered in exogenous
anaerobic infections or intoxications.
 Its toxins usually gain access to the body through
ingestion or via open wounds that have been
contaminated with soil.
 Contribute to virulence: collagenase,
hyaluronidase, lecithinase (cell destruction) and
phospholipase
 Histotoxic clostridia (myonecrosis): C.
perfringens, C. novyi, C. septicum, C.
histolyticum and C. bifermetans
 C. septicum - is a marker for a malignancy in the
GI tract; "smooth swarming" growth on plated
media.

General Characteristics of Clostridia:

 They formed endospores anaerobically.
 They are motile with peritrichous flagella except
C. perfringens, C. ranosum and C. innocum.
 They have swollen sporangia except C.
perfringens and C. bifermentans. They are non-
encapsulated except C. perfringens.
 They have a single hemolytic reaction except C.
perfringens.
Clostridium tetani (Tack head bacillus)
 Is a soil and environmental inhabitants.
 The endospores are found in hospital
environments, in soil and dust, and in the feces of
many farm animals.
 Virulence factor: tetanospasmin (neurotoxin)
 Microscopy: with terminal spore and swollen
sporangia - "drumstick/lollipop/tennis
 racket" appearance
 Cuiture: BAP heavy "smooth swarming"
anaerobically but grow slowly
BAP-colonies are with matte surface with
narrow zone of B-hemolysis
 Biochemical test: motile; (+) gelatinase and
indole; (-) lecithinase and lipase
Tetanospasmin
 Is an endopeptidase that selectively cleaves the
synaptic vesicle membrane protein
synaptobrevin.
 It causes tension or cramping and twisting in
skeletal muscles surrounding the wound and
tightness of the jaw muscles.
Associated Disease:
Tetanus
 It is characterized by "trismus" (lockjaw) and
"risus sardonicus" (distorted grin).
 It occurs when the the organism (spore) enters an
open wound and elaborates the potent toxin that
mediates generalized muscle spasms.
 Symptoms: muscular rigidity (jaws, neck and
lumbar region), difficulty in swallowing, rigidity
of the abdomen, chest, back and limbs Incubation
period: 3 to 21 days (the long incubation period
is related to the distance from the injury to the
CNS)
 Tetanus neonatorum is caused by contaminated
instruments used for newborns
Clostridium botulinum (Canned good bacillus)
 It is found in soil and aquatic sediments.
 It is a potential agent of bioterrorism.
 It is characterized by the presence of subterminal
spore; B-hemolytic on BAP.
 Is a proteolytic bacterium with the ability to
oxidize one amino acid, with the second amino
acid as the electron acceptor (Stickland reaction).
 Virulence factor: botulism toxin- neurotoxin
(most potent toxin known to man)
Botulism toxin
 It selectively cleaves the synaptic vesicle
membrane protein synaptobrevin, thus
preventing exocytosis and release of the
neurotransmitter acetylcholine.
 It takes only a small amount of botulism toxin to
produce paralysis and death.
 Botulism toxin has 7 antigenically different types
(A to G) only types A, B and E are associated
with human infections.
 Taxin type A is used medically to treat
strabismus (wandering eye) and as a beauty
enhancer by temporarily improving frown lines
(botox treatment).
Clinical Infection:
Botulism
 - Main feature: double or blurred vision,
impaired speech, difficulty in swallowing,
1. Foodborne botulism
 weakness and paralysis
 It results from ingestion of preformed toxin in
nonacidic vegetable, preserved food, nieat-based
food or mushroom foodstuffs.
 It is commonly caused by botulism toxin A.
2. Infant botulism
 Is an actual infection caused by ingesting the
organism from honey or via breast feeding.
Clostridium difficile
 It is the most common cause of antibiotic-
associated diarrhea and pseudomembranous
colitis (bloody diarrhea with necrosis of colonic
mucosa).
 It is acquired in the hospitals by individuals
receiving antibiotics.
 It is found as part of the GI biota in about 5% of
individuals.
 It is an "infection control dilemma" among
hospitalized patients
 LISTERIORIS
Is a serious infection primarily of neonates, pregnant
women and immunocompromised
hosts.

ERYSIPELOTHRIX RHUSIOPATHIAE
 Erysipelothrix means "red skin, thread";
rhusiopathiae means "red disease."
 The only species in the genus that cause human
disease; not part of the human flora. The only
catalase negative, Gram-positive non-
LISTERIA
sporeforming rod that produces H2S.
LISTERIA MONOCYTOGENES
 It is aerobic or facultatively anaerobic; non
 It is both human and animal pathogen; also
motile, nonhemolytic; non sporeformer.
isolated from crustaceans, ticks and flies. It is
 It is transmitted to humans from animals by
recovered from soil, dust, water, sewage, silage,
contact with infected animal excreta, blood and
dairy products and processed meats. It is motile
flesh (open skin wounds).
with the characteristic "tumbling motility".
 It is widespread in wild and domestic animals -
 It is aerobic or facultatively anaerobic, non-
birds and fish.
sporeforming; (+) catalase.
 It is resistant to salting, pickling and smoking;
 It can grow in high salt concentration (up to 10%
survives well in soil and water. Microscopy: thin,
NaCl).
pleomorphic rod; it has tendency to form long
 It has the ability to survive within phagocytes
filaments arranged in singly, short chains or in a
(intracellular pathogen). It has optimal growth
V-shape
between 30°C and 37°C but also grows at 4°C.
 Gelatin stab culture: "pipe cleaner" or "test tube
Virulence factors: listcriolysin O (hemolytic and
brush" pattern at 22°C
cytotoxic), catalase, SOD, phospholipase C and
 Major reservoir: domestic swine
p60 (surface protein)
Clinical Infections:
 Microscopy: Gram-positive coccobacillus, in
 Human infections result from occupational
singly or short chains resembling streptococci;
exposure fish handlers and animal products,
may appear as Corynebacteria when the bacillus
abattoir workers and veterinarians.
forms prevail
 It is transmitted to humans from animals by
 Culture: BAP-small, smooth, translucent grayish-
means of skin trauma (open wound).
blue colonies surrounded by a narrow zone of B-
hemolysis

Clinical Infections:
 Ingestion of contaminated food such as meat,
chicken, dairy products and coleslaw.
3 Types of Infections:
 Left-over or ready-to-eat foods such as hotdogs
1. Septicemia
or cold cuts should be thoroughly heated before
2. Endocarditis individual with valve replacements
consumption.
3. Erysipeloid
 Individuals who are immunocompromised and
 Is a localized skin infection that resembles
pregnant women should avoid eating soft cheeses
streptococcal erysipelas.
such as mexican-style, feta, brie, camembert and
 It is a self-limiting infection that normally heals
blue-veined.
within 3-4 weeks; relapses are common. Lesions
 Colonized mothers may pass organism onto
are seen on the hands or fingers through
fetus.
inoculation of organism.
 Macrophages and T lymphocytes are the most
 Signs and symptoms: painful, swollen, slightly
important host defenses against infection.
elevated, spreading purplish-red zone
CORYNEBACTERIUM
 • Majority of the species are found as normal
flora on skin and mucous membranes of humans
and animals.
 Are animal, plant and human pathogens.
 Are found in fresh and salt water, soil and air.
 They are non-motile, non-sporeforming, non-
encapsulated; pleomorphic rods. Are closely
related to Mycobacteria and Nocardia - the cell
walls of Corynebacteria contain
mesodiaminopimelic acid (m-DAP) as the
diamino acid and short-chain mycolic acids.
 Are glucose and maltose fermenters except C.
urealyticum and C. pseudodiphtheriticum.
 Microscopic: slightly curved Gram-positive rods
with unparallel sides and slightly wider ends -
"club shape bacteria"
 Biochemical test: (+) catalase and oxidase
 Growth factors: 8 amino acids
PATHOGENESIS:
DIPHTHERIA
 Is an acute, contagious disease characterized by
the production of a systemic toxin and a false
membrane lining (pseudomembranous
formation) of the mucous membrane of the throat
leading to respiratory obstruction.
 The only effective control is through
immunization (DPT) diphtheria antitoxin is
administered to neutralize any unabsorbed
exotoxin in the patient's tissues.
Diptheria toxin
 It is produced by strains infected with a
lysogenic B-phage, which carries the tox gene.
 It inhibits protein synthesis in eukaryotic cells.
 It is secreted by the bacterial cell and is nontoxic
until exposed to trypsin "trypsinazation" cleaves
the toxin into 2 fragments (Fragment A and B).

3. LACTOBACILLUS ACIDOPHILUS/
DODERLAIN BACILLi
- produced large amount of lactic acid, non
pathogenic and has little clinical significance-
Tomato Juice agar
• Normal flora of mouth, GIT and vaginal canal

CORYNEBACTERIUM DIPHTHERIAE
 Diphtheria bacillus/Kleb Loeffler's bacillus
 Is aerobic or facultative anaerobe; pleomorphic
rods but grow best under aerobic conditions.
 It inhabits human nasopharynx but only in carrier
state; not part of the normal flora of respiratory
tract.
 It is acquired through contaminated respiratory
droplets or direct contact with infected cutaneous
lesions (hand-to-mouth) - humans are the only
host.
 Enriched medium is recommended (serum,
cystine and K tellurite) for growth; multiplication
also occurs at 15-40°C.
 It is readily killed by heat and most of the usual
disinfectants.
 It is very resistant to drying and remains viable in
the environment for weeks. Biochemical test: (+)
catalase; (-) urease; reduces nitrate to nitrite
produced acid but not gas ;nonmotile 2 Forms of Diphtheria:
 Virulence factor: diphtheria toxin (130ng/kg 1. Respiratory form
BW-lethal dose)
 Pharyngitis is characterized by the development
of an exudative membrane that covers the tonsils,
uvula, palate and pharyngeal wall.
 It is acquired thru droplet infection or hand-to-
mouth contact.
 It mainly affects people living in crowded
environment.
 Tissue necrosis and exudate formation: gray to
white pseudomembranous formation
 Extension of the pseudomembrane superiorly
into the nasopharynx or inferiorly into the larynx
may be so marked to produce respiratory
obstruction.
 Death is often a result of cardiac failure.
 Signs and symptoms: low-grade fever, thick
mucupurulent nasal discharge and cough.
2. Cutaneous form (Velat sore/Barcoo rot)
 Is characterized slow-healing ulcers and
membrane formation.
 Toxin is absorbed systematically but
complication is less common.
Corynebacterium diphtheriae Group
 Members: C. diptheriae, C. ulcerans and C.
pseudotuberculosis
Laboratory Diagnosis:
Specimen: nasopharyngeal and throat swabs
 Calcium alginate swab is preferred for collection.
1. Gram Stain
 Highly pleomorphic Gram-positive short or
slightly curved rods with rounded ends.
 Cells may also appear in club-shaped swellings
and beaded forms (when stained with methylene
blue).
 Cells are arranged in palisades or in pairs
forming X, V, Y & L formations resembling
"chinese letters" - because of incomplete fission
during multiplication.
 Stains for metachromatic granules: LAMB,
Neisser's, Albert's and Ljubinsky stains
2. Culture
 Culture media: BAP, CAP, CTBA, Tinsdale
Agar and Loeffler Serum Agar
 BAP-small zone of ẞ hemolysis; some are non
hemolytic.
 Primary inoculation of throat swabs to a Loeffer
slant and overnight incubation and subculture of
any growth to cystine-tellurite blood agar may
improve recovery of C. Diptheriae.
Corynebacterium ulcerans
 Is an animal pathogen causing mastitis in cattle
and other domestic animals.
 It is acquired through contact with animals or
ingestion of unpasteurized dairy products.
 It has been associated with diphtheria-like sore
throat.
 A significant number of isolates produce the
diphtheria-like toxin.
 It has been isolated also from skin ulcers and
exudative pharyngitis.
 (+) gelatin formation at room temperature; does
not reduce nitrate.
 Culture: BAP-narrow zone of B-hemolysis
CTBA-(+) brown halo around colonies grows well
on Loeffler's agar
Corynebacterium pseudotuberculosis
 Is an animal pathogen-humans acquired infection
through animal contact.
 It causes a granulomatous lymphadenitis in
humans.
 It produces a dermonecrotic toxin that causes
death of a variety of cell types.
 It can also produce diphtheria toxin.
 Culture: brown halo on CTBA; small, yellowish-
white colonies on BAP Biochemical test: (+)
urease; (-) gelatin formation
Corynebacterium jeikeium
 Is a common skin flora of hospitalized
individuals (inguinal, axillary and rectal sites).
 It is a lipophilic species; obligate aerobe; (+)
urease and reduce nitrate.
 It is isolated from immunocompromised
individuals, have undergone invasive procedures,
or have a history of IV drug abuse.
 It is a common cause of diphtheroid prosthetic
valve endocarditis in adults.
 It is a multiple antibiotic resistant (ẞ lactam)
bacterium which typically survived in a hospital
setting.
■ Microscopy: pleomorphic, club-shaped, and
arranged in V forms
⚫ Culture: BAP - nonhemolytic; appear as large
colonies when supplemented
with 1% Tween 80
Corynebacterium urealyticum
 Is one of the most frequently isolated clinically
significant corynebacteria.
 It is a urinary pathogen; strict aerobe; lipophilic.
 It has shown resistance to several antibiotics such
as B-lactams.
Nocardia
 They are aerobic Gram-positive bacilli with long,
thin, beaded, branching filaments that
occasionally fragment into rod-shaped and short
coccoid forms during aging.
 Is partially acid-fast, strictly aerobic, catalase (+)
bacilli.
 The cell wall contains peptidoglycan,
mesodiaminopimelic acid (DAP), and the sugars
arabinose and galactose.
 It grows on nonselective media but it may take a
week or more.
 It also grows on media used to recover fungi.
 Extreme variable colonial growth: wrinkled, dry,
chalky-white to orange-tan pigment colonies
("breadcrumbs-looking") or can be waxy, bumpy
or velvety rugose forms with yellow to orange
pigment in 5-20 days
 Species: N. asteroides, N. brasiliensis, N.
farcinica and N. nova
 Differential test: resistant to lysozyme
Nocardia asteroides complex
 They are responsible for 80-90% of human
infections.
 They are facultatively intracellular parasites -
resistant to intracellular killing and cause tropism
for neuronal tissue.
 They have the ability to inhibit phagosome-
iysosome fusion.
 They produce large amounts of catalase and
hemolysin.
 Species: N. cyriageorgica, N. brasiliensis, N.
otitidis caviarum
Clinical Infections:
 Infection is acquired through inhalation of the
organism present in dust and soil.
 It can cause invasive pulmonary infections.
 Nocardia can spread hematogenously throughout
the body from a primary pulmonary infection.
 Major causes of infections: N. asteroides and N.
brasiliensis
 N. asteroides is the common cause of pulmonary
disease.
 N. brasiliensis is the predominant cause of skin
infections.
Rhodococcus, Gordona, Tsukamurella
 Gram-positive aerobic, catalase (+), branching,
filamentous bacteria that can fragment into rods
and cocci.
 They can be isolated from soil, fresh and marine
water, and organic matter.
 They are primarily acquired by inhalation.
 They can grow on most of the nonselective
media for bacterial, mycobacterial and fungi
isolation.
 Infections: skin infections, pneumonia,
perotinitis, catheter-associated sepsis
HACEK GROUP
 Haemophilus spp- Aggregatibacter aphrophilus
(formerly H. aphrophilus
Aggregatibacter actinomycetemcomitans
Cardiobacterium hominis
Eikenella corodens
 Kingella spp (K. kingae-HACEK species)
 ■ Are small, nonmotile, nonsporeforming, Gram-
negative bacilli that will not grow on Mac
Conkey agar.
 Are part of the normal oral flora; opportunistic
pathogens.
 Are fastidious and require increased CO2 at least
for initial isolation from clinical specimens.
 They have slow growth (dysgonic) on BAP and
CAP (7-14 days).
 They require an additional 1 to 2 days before
they can be isolated from blood cultures.
 They caused slow, progressive bacterial
endocarditis ("vegetation").
 They utilized 6-aminolevulinic acid; all are (-)
indole except C. hominis.
AEROBIC ACTINOMYCETES
 They are large and diverse group of Gram-
positive bacilli.
 They are aerobic, branched and beaded - the
branched filaments that extend along the agar are
called substrate hyphae, and into the agar are
aerial hyphae.
 They are found in soil and organic material.
 They cause disease in many animals and organic
material.
 Some have ability to grow at 50°C like the
thermophilic actinomycetes.
 They have the ability to degrade amyl alcohol,
paraffin and rubber.
 Culture: the cells elongate to form branching,
filamentous forms some organisms form
filaments or hyphae on the agar surface or into
the agar.
3 Types of Skin Infections:
1. Actinomycetoma - chronic, localized, painless
subcutaneous infection; presence of "sulfur granules"
2. Lymphocutaneous infection
3. Pulmonary disease-confluent bronchopneumonia;
sputum is thick and purulent; no encapsulation of the
abscess; no sulfur granules
Laboratory Diagnosis
Specimens: biopsy or drainage material from
actinomycetoma, sputum,
bronchoalveolar lavage, lung tissue, transthoracic
aspirate of a nodule or abscess, CSF and blood.
SMALL PLEOMORPHIC GRAM-NEGATIVE
BACILLI
HAEMOPHILUS
 The genus name is derived from the Greek
words meaning "blood lover".
 They normally inhabit the URT of humans
except H. ducreyi - in adults it is mostly consists
of H. parainfluenzae.
 Are obligate parasites on the mucous membranes
of humans.
 Are fastidious and nonmotile; nonsporeforming;
capnophilic and facultatively anaerobic bacteria.
 They die rapidly in clinical specimen - very
susceptible to drying and extreme temperature.
 Most species will not grow on pure BAP
Microscopy: Gram-negative, small, pleomorphic,
coccobacilli or rods
 Culture: BAP-moist, smooth, convex colonies on
agar plate
Biochemical test: catalase (+); oxidase
(+) except H. segnis.
 Growth factors: X (hemin) and V (NAD)
 Human pathogens: H. influenzae, H. ducreyi, H.
parainfluenzae, H. paraphrophilus, H.
parahaemolyticus, H. pittmaniae, H. aegypticus,
and H. segnis
Haemophilus influenzae
 It is transmitted by person to person by
contaminated respiratory droplets.
 It may be mistaken for S. pyogenes if the Gram
stain is not well decolorize.
 It is the commonly tested organism for B-
lactamase production.
 It is the main cause of meningitis in children
nasopharynx to the regional lymph nodes to the
blood and finally to meninges. the organism
spreads from the The only member of the genus
that produce IgA protease.
 Do not produce endotoxin; rapidly killed by
phagocytes; very fastidious.
 Not all strains are encapsulated microbiota of the  Chancroid-genital lesions; from tender papules
URT. nonencapsulated strains are part of the to painful ulcers with several satellite lesions.
normal
 Culture: CAP-translucent, convex, tan mucoid Haemophilus aegypticus (Koch-Weeks bacillus)
colonies  Is genetically related to H. influenzae.
"mousy or bleach-like odor"  It was observed in conjunctivitis exudates from
Egyptians by Koch in 1883.
Principal virulence factor: polysaccharide capsule  It is the etiologic agent of pinkeye conjunctivitis.
(serotypes a-f) Other virulence factors: IgA proteases,
fimbriae, OMPs and LPS Haemophilus influenzae biogroup aegypticus
 (-) Porphyrin test- this test detects the presence (nonencapsulated)
of enzymes that converts & aminolevulinic acid  It causes conjunctivitis primarily in pediatric
(ALA) into porphyrins. populations.
 It is the etiologic agent of Brazilian purpuric
2 Categories: fever (BPF).
a. Typeable based on capsular characteristics
 Types a,b,c,d,e,f (capsular serotypes)-
encapsulated strains
Type b is the cause of serious infections in humans

b. Nontypeable
 Are normal inhabitants of URT-adheres to
human epithelial cells.
 Do not produce capsule (non encapsulated
strains).
 It causes otitis media with effusion (middle ear
infection) - the 2nd prevalent etiologic
individuals with underlying agent after S.
pneumoniae.
 It also causes pneumonia in elderly patients and
 respiratory infections including cystic fibrosis.
 Other related infections: conjunctivitis and
sinusitis (localized infections)

Haemophilus ducreyi
 Is not part of human flora; only found in humans
during infection.
 It infects mucosal epithelium, genital and
nongenital skin, and regional lymph nodes.
 Is the agent of chancroid or "soft chancre" - a
highly communicable sexually transmitted
 genital ulcer disease.
 Suppurative, enlarged, draining, inguinal lymph
nodes (buboes) are common in the majority of
infected patients.
 It is commonly seen in socioeconomically
disadvantaged population.
 Culture: CAP transparent, small, flat, smooth,
nonmucoid, tan or yellow colonies Saline
suspension difficult to pick up; "clumpy"
appearance