Topics:

Pseudomonas Aeruginosa, Legionella,

Brucella, Bordetella, and Francisella
 Pseudomonads
• Gram-negative, aerobic,
rod-shaped bacteria

• The most important

species from a medical
point of view (production
of enzyme)
OCCURRENCE:

• Abundant in the environment

• Causes infections in persons with immune

deficiency
Form and structure:

• plump, 2–4 um long rods with one to several polar flagella

• can produce a viscous extracellular slime layer
Form and structure:
• P. aeruginosa possesses an outer membrane as part of its
cell wall. The architecture of this membrane is responsible for the
natural resistance of this bacterium to many antibiotics.

• P. aeruginosa can only be grown in culture mediums

Culture

• P. Aeruginosa can only be grown in culture mediums containing free

O2 as a terminal electron acceptor (Obligate aerobe).
• In nutrient broth, the organism grows at the surface to form a so-
called pellicle.
Colonies on nutrient
agar
P. aeruginosa can
produce two pigments,
i.e., both yellow-green
fluorescein
and blue-green
pyocyanin.
 Pathogenesis and clinical pictures

• P. aeruginosa
usually enters
body tissues
through injuries

• attaches to tissue
cells using specific
attachment
fimbriae.
• The most important virulence factor is exotoxin A
(ADP ribosyl transferase), which blocks translation in
protein synthesis by inactivating the elongation factor
eEF2.
• The exoenzyme S (also an ADP ribosyl transferase)
inactivates cytoskeletal proteins and GTP-binding
proteins in eukaryotic cells.
P. AERUGINOSA INFECTIONS:
• pneumonias in cystic fibrosis or in
patients on respiratory equipment,
• infections of burn wounds,
• postoperative wound infections,
• chronic pyelonephritis,
• endocarditis in drug addicts,
• sepsis,
• malignant otitis externa.

*P. aeruginosa frequently causes

nosocomial infections
Diagnosis:

• Laboratory diagnosis includes isolation of the

pathogen from relevant materials and its
identification based on a specific pattern of
metabolic properties.
Epidemiology and prevention:

Pseudomonas aeruginosa is considered a hospital

problem due to:

1. its antibiotic resistance

2. opportunistic nature
3. presence in medical devices and water sources
4. potential for patient-to-patient transmission
5. propensity for causing outbreaks in healthcare settings.
Proliferate in moist condition such as:

• Sinks
• toilets
• cosmetics
• vaporizers
• inhalers
• respirators
• anesthesiology equipment
• dialysis equipment
Therapy
Antibiotics that can be used to treat P. aeruginosa

• aminoglycosides,
• acylureidopenicillins,
• carboxylpenicillins,
• group 3b cephalosporins
• carbapenems.
• Combination of an aminoglycoside
with a betalactam is indicated in severe infections.

Susceptibility tests are necessary due to frequent resistance.

Other Pseudomonas species
• Burkholderia mallei - This species is the causative
organism in malleus orglanders, a disease of
solipeds.

• Burkholderia pseudomallei. - Starting from primary

foci, the infection can disseminate and cause
abscesses in a number of different
organs. This disease is observed mainly in Asia.
 Summary: Pseudomonas
Aeruginosa
Legionella (Legionnaire’s Disease)

• Legionella is the
only genus in the
family
Legionellaceae.
• Legionella
pneumophila is
responsible for most
legionelloses in
humans.
• Legionellae are
difficult to stain.
Legionnaire’s Disease

• They are Gram-negative, aerobic rod bacteria.

• Special mediums must be used to grow them in cultures.
• Legionalla occur when droplets containing the pathogens are
inhaled.
• The persons most likely to contract legionnaire’s disease are
those with a primary cardiopulmonary disease and generally
weakened immune defenses.
Laboratory diagnostic method:

• microscopy with direct

immunofluorescence

• culturing on special
mediums and
antibody assays
Other information:

• Antibiotics of choice are macrolides

• Sources of infection include hot and cold water supply system
• Cooling towers
• Moisturizing units in air conditioner
• Whirpool baths
Epidemiology and prevention.

• Legionellosis can occur in epidemic form or in sporadic

infections.
• It is estimated that one third of all pneumonias requiring
hospitalization are legionelloses.
• Soil and damp biotopes are the natural habitat of Legionella.
• Human-to-human transmission has not been confirmed.
• Legionella bacteria tolerate water temperatures as high as 50
C and are not killed until the water is briefly heated to 70 C.
Brucella, Bordetella, and Francisella

• small, coccoid, Gram-negative rods

• can be cultured under strict aerobic

conditions on enriched nutrient mediums.
Brucellosis – Mediterranean
fever
• a classic zoonosis that
affects cattle, goats, and
pigs

• pathogens can be
transmitted to humans
directly from diseased
animals or indirectly in
food

• primary clinical symptom

is the undulant fever
 Brucella (Brucellosis, Bang’s Disease)
• The genus Brucella
includes three medically
relevant species—

1. B. abortus,
2. B. melitensis,
3. B. suis

Can be transmitted from

animals to humans causing
”Bang’s Disease”
Occurrence and classification.
• The genus Brucella includes three medically relevant species
—B. abortus, B. melitensis, and B. suis.

• These three species are the causative organisms of classic

zoonoses in livestock and wild animals, specifically in cattle
(B. abortus), goats (B. melitensis), and pigs (B. suis). These
bacteria can also be transmitted from diseased animals to
humans, causing a uniform clinical picture, so-called
undulant fever or Bang’s disease.
Morphology and culture.
• Brucellae are slight, coccoid,
Gram-negative rods with no
flagella.
• only reproduce aerobically.
• In the initial isolation the
atmosphere
• must contain 5–10% CO2.
• Enriched mediums such as
blood agar are required to
grow them in cultures.
Pathogenesis
Infections result from:

1. direct contact with diseased

animals
2. indirectly by way of
contaminated foods particular
unpasteurized milk and dairy
products
Brucellosis Transmission:
The bacteria invade the body either through:

• the mucosa of the upper intestinal

• respiratory tracts or through lesions in the skin
• then enter the subserosa or subcutis.
• transported by microphages or macrophages, in
which they can survive, to the lymph nodes.
Diagnosis

•Achieved by isolating the pathogen

from blood or biopsies in cultures,
which must be incubated for up to
four weeks
Therapy
• Doxycycline is administered in
the acute phase, often in
combination with gentamicin

• A therapeutic alternative is
cotrimoxazole.

• The antibiotic regimen must be

continued for three to four
weeks.
Bordetella
3 Pathogens responsible for whooping
cough
• The genus Bordetella, among others, includes the species

1. B. pertussis – greatest concern for humans

2. B. parapertussis,
3. and B. bronchiseptica.
Form Structure and culture

• small, coccoid, nonmotile, Gram-

negative rods

• Grow aerobically on special

culture mediums at 37 8C for three
to four days.
 Morphology and Culture

• B. pertussis bacteria are small, coccoid, nonmotile,

Gram-negative rods that can be grown aerobically on special culture mediums
at 37 C for three to four days.
 Bordetella
• Whooping cough is an
infectious disease also called
Pertussis.

• infection of the airways and

the mucus layers lining it.

• affects the windpipe (trachea)

and the branches of the
windpipe or the bronchi.
 Bordetella (Whooping Cough, Pertussis)

Characteristic:

• typical hacking
cough
• sharp intake
of breath
• sounds like a
“whoop”.
Pathogenesis
Pertussis bacteria
are transmitted by
aerosol droplets.

-attach themselves
to the cells of the
ciliated epithelium
in the bronchi.
Pathogenesis

They rarely invade

the epithelium. The
infection results in
(sub-) epithelial
inflammations and
necroses.
Clinical picture.

• The onset of whooping cough (pertussis) develops after an

incubation period of about 10–14 days with an
uncharacteristic catarrha phase lasting 1–2 weeks

• followed by the two to three week-long paroxysmal phase

with typical convulsive coughing spells.
• Then comes the convalescent phase, which can last for several weeks

• Frequent complications, especially in infants, include secondary

pneumonias caused by pneumococci or Haemophilus, which are able
to penetrate readily through the damaged mucosa, and otitis media.
Diagnosis:
• The pathogen can only be isolated and identified during the catarrhal
and early paroxysmal phases.
• Specimen material is taken from the nasopharynx through the nose using a
special swabbing technique.
• A special medium is then carefully inoculated or the specimen is transported
to the laboratory using a suitable transport medium.
• B. pertussis can also be identified in nasopharyngeal secretion using the
direct immunofluorescence technique. Cultures must be aerobically
incubated for three to four days. Antibodies cannot be detected by EIA until
two weeks after onset at the earliest.
• Only a seroconversion is conclusive.
Therapy.

• Antibiotic treatment can only be expected to be effective

during the catarrhal and early paroxysmal phases before the
virulence factors are bound to the corresponding cell
receptors. Macrolides are the agents of choice.
Antibiotics: macrolides
Epidemiology and prevention

• Pertussis occurs worldwide. Humans are the only hosts. Sources of

infection are infected persons during the catarrhal
phase, who cough out the pathogens in droplets. There are no healthy
carriers.

• The most important preventive measure is the active vaccination (see

vaccination
schedule, p. 33). Although a whole-cell vaccine is available, various
acellular vaccines are now preferred.
 Pathogenesis.

• Pertussis bacteria are transmitted by aerosol droplets.

• attach themselves to the cells of the ciliated epithelium in the
bronchi.
• rarely invade the epithelium.
• infection results in (sub-) epithelial inflammations and necroses.
Review:

• The causative organism of

whooping cough, which affects
only humans.

• Pathogens are transmitted by

aerosol droplets.

• The most important prophylactic

measure is the vaccination in
the first year of life.
Francisella tularensis bacteria (Tularemia)
 Francisella tularensis Bacteria (Tularemia)

• coccoid, nonmotile, Gram-negative,

aerobic rods.

• cause a disease similar to plague in

numerous animal species, above all in
rodents.

• Humans are infected by contact with

diseased animals or ectoparasites or dust.
 Francisella tularensis Bacteria (Tularemia)
• An ulcerous lesion develops at the portal of
entry that also affects the local lymph nodes
(ulceroglandular, glandular, or oculoglandular
form).
• Via lymphogenous and hematogenous
dissemination the pathogens then spread to
parenchymatous organs, spleen and liver.
• Small granulomas develop, which develop
central caseation or purulent abscesses.
• usually affects animals like rabbits, rodents ,rats, squirrels or birds,
domestic animals but may spread to humans via animal bite or direct
exposure through any other way.
• highly contagious disease but can be managed by medications.
• In research it is evident that this bacteria was used as a biological weapon
• The incubation period is three to four days.
• Diagnostic procedures aim to isolate and identify the pathogen in
cultures and under the microscope.
• Agglutinating antibodies can be detected beginning with the second
week.
• A seroconversion is the confirming factor.
• Antibiosis is carried out with streptomycin or gentamicin.
Symptoms Of Tularemia

• Headache, exhaustion, fever are the basic symptoms.

• Specific symptoms are also there on the basis of different types of

tularemia.

• People exposed to this problem may get sick within three to five days
or max 21 days.
Therapy
Antibiosis

• streptomycin or
gentamicin.