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Zoonosis
Francisella
Brucella
Yersinia
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ZOONOSIS
A disease, primarily of animals, which
is transmitted to humans as a result ofdirect or indirect contact with the
infected animal population
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Brucellosis
1. Overview
2. Morphology &
Physiology
3. Epidemiology
4. Symptoms
5. Pathogenesis
6. Diagnosis
7. Treatment
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Brucella: Overview
Primarily a disease of animals.
Common where significant disease among
domestic animals.
Common names- Undulant fever, Maltafever, Mediterranean remittent fever.
Brucella can go through intact skin.
Facultative intracellular bacteria
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Morphology & Physiology
Small gram-negativecoccobacillus
Grows slowly (7 days), at 370 C.
On subculture, a minimum of 48 h growth Aerobic growth on Chocolate agar and Sheep
blood agar
Will not grow on MacConkey or Eosin methyleneblue (EMB) agar
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Morphology & Physiology
Non-pigmented and non-hemolytic
Non-motile
Oxidase: positive
Catalase: positive
Urease: strongly positive, less than 2 hours. Some
species within 5 minutes.
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Microscopic Characteristics
Brucellaspp.
poorly staining
small gram-negativecoccobacilli
seen mostly as single cells
appearing like fine sand
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Brucella melitensiscolonies
A. Grows slowly on
most standard
laboratory media.
Usually not visible
at 24h.
B. Pinpoint,
smooth,
translucent, non-
hemolytic at 48h.
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Public Health Aspects
Brucella: Sources
Brucellosis caused by 1 of 4Brucellaspecies:
1. B. abortus
Some strainsrequire 5% CO2on initial
isolation.
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2. B. melitenus
Goats
Sheep
Camels
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3. B. su is
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4. B. canis
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2 patient populations
1. Individuals who work
with unvaccinated
animals
B. abortus and B. suis
Infections result from:
direct contact inhalation
2. Individuals who ingest
unpasteurized dairy
products
B. melitensis is the most
common agent
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Host Animal - Brucellosis
Asymptomatic or mild disease.
Predilection for organs rich in erythritol (breast,uterus, placenta, epididymis).
Causes sterility, abortions or carrier state in non-
human animals.
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Human - Brucellosis
Symptoms
AcutePhase AdvancedDisease
ChronicForm
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Organism Animal
reservoir
Human disease Complications
B. abortus Cattle mild suppurative febrile
infection
Rare
B. canis Dogs mild suppurative febrileinfection Rare
B. suis Swine Prolonged disease with
destructive lesions of the
lymphoreticular organs and
kidney
B. melitensis goats/sheep Severe and recurring disease High incidence of
serious
complications
Human - Brucellosis
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Pathogenesis
mucosal epithelium
Transported to lymph nodes,spleen, liver and bone marrow.
Brucella
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Pathogenesis
Phagosome
Lysozome
X
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Pathogenesis
No exotoxin
LPS does not activate the alternativecomplement pathway
Acute lymphadenitis
Granulocyte production in lymphatic tissue,
spleen, liver, bone marrow, lymph nodes andkidneys.
A potential bioterrorist agent
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Treatment
Tetracycline, doxycycline, or
trimethoprimsulfamethoxazole in combination
and rifampin or gentamicin for 6 weeks to preventreoccurring infection.
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Tularemia:
(Franc isel la tularens is)
Gram stain
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Tularemia: Overview
Primary reservoir in US Rabbits and muskrats
Insect vectors
Ticks
Infection via Insect bites
Handling contaminated animal tissues
Inhalation of aerosols
Ingestion of contaminated food or water
Exposure in a laboratory setting
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Tularemia: Overview
Gram-negative coccobacilli.
Low infectious dose
Two subspecies of F . tularensis:
subspecies tularensis (type A)
subspecies holarctica (type B)
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Morphology & Physiology
Tiny gram-negative coccobacillus
Nonmotile, encapsulated
Aerobic slow growing (48 hours) 35-370C
Fastidious organism requires sulfhydryl (cysteine,IsoVitaleX) supplementation for growth
Grows wells on
Chocolate agar Buffered charcoal yeast extract agar
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Colony Characteristics
After 48 hours incubation
Colonies
Very small
white to gray to bluish-
gray
Will not grow on
MacConkey or EMB plates.
F. tularensis on chocolate agar 48 hours growth.
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Microscopic Characteristics
Tiny, faintly staining, pleomorphic gram-negative rods
(0.2-0.5 mcm X 0.7-1.0 mcm) are noted; cells are smallerthan those of Haemophiluss ecies.
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Grows slowly at 35-370 C
Oxidase-negative
Weakly catalase-positive (may be negative) Urea-negative
Nitrate-negative
Non-motile
Beta-lactamase-positive
Satellite or XV test-negative (unlike Haemophilus)
Phenotypic Characteristics
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Tularemia: Public Health
Modes of humans infection
Bite of infected flies, or ticks
Handling contaminated animal tissues or fluids
Direct contact with or ingestion of contaminatedwater, food, or soil
Inhalation of infective aerosols (most likely BT
route)
http://images.google.com/imgres?imgurl=http://insects.tamu.edu/images/animalia/arthropoda/insecta/diptera/tabanidae/chrysops_unknown_adult_dorsal_m_01.jpg&imgrefurl=http://insects.tamu.edu/extension/youth/bug/bug142.html&usg=__KMPZ_azUJI0Wcb4g3GnOTrOyh0M=&h=423&w=640&sz=26&hl=en&start=1&um=1&tbnid=Y4T9ZV9OZAqkpM:&tbnh=91&tbnw=137&prev=/images%3Fq%3Ddeer%2Bflies%26hl%3Den%26rls%3Dcom.microsoft:en-us:IE-SearchBox%26rlz%3D1I7ADBR_en%26um%3D1http://images.google.com/imgres?imgurl=http://insects.tamu.edu/images/animalia/arthropoda/insecta/diptera/tabanidae/chrysops_unknown_adult_dorsal_m_01.jpg&imgrefurl=http://insects.tamu.edu/extension/youth/bug/bug142.html&usg=__KMPZ_azUJI0Wcb4g3GnOTrOyh0M=&h=423&w=640&sz=26&hl=en&start=1&um=1&tbnid=Y4T9ZV9OZAqkpM:&tbnh=91&tbnw=137&prev=/images%3Fq%3Ddeer%2Bflies%26hl%3Den%26rls%3Dcom.microsoft:en-us:IE-SearchBox%26rlz%3D1I7ADBR_en%26um%3D1http://images.google.com/imgres?imgurl=http://insects.tamu.edu/images/animalia/arthropoda/insecta/diptera/tabanidae/chrysops_unknown_adult_dorsal_m_01.jpg&imgrefurl=http://insects.tamu.edu/extension/youth/bug/bug142.html&usg=__KMPZ_azUJI0Wcb4g3GnOTrOyh0M=&h=423&w=640&sz=26&hl=en&start=1&um=1&tbnid=Y4T9ZV9OZAqkpM:&tbnh=91&tbnw=137&prev=/images%3Fq%3Ddeer%2Bflies%26hl%3Den%26rls%3Dcom.microsoft:en-us:IE-SearchBox%26rlz%3D1I7ADBR_en%26um%3D1 -
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Endemic in US
Majority of cases occur MaySeptember (tick
exposure) or winter (hunters).
Most in rural areas.
Arkansas, Missouri and Oklahoma
Tularemia: Public Health
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Symptoms
Incubation period: 3-5 days (range 1-21 days)
Clinical presentation can be divided into groups
Ulceroglandular (45-85%) /glandular (10% to 25%)
Typhoidal
Pneumonic
Oculoglandular
Oropharyngeal/Gastrointestinal
Prominent lymphadenopathy
Recovery followed by permanent immunity
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Tularemia Clinical Types
Clinical presentation based on theroute of infection
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Ulceroglandular & Glandular tularemia
Ulceroglandular accounts for 75-85% of naturally occurring cases.
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Typhoidal tularemia
Bacteremia- Sepsis
Fever, chills, headache, myalgias, malaise,
sore throat, and anorexia.
Likely bioterrorism presentation.
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Pneumonic tularemia
Entry into lungs via
Aerosols
hematogenous
Severe atypical pneumonia
Likely BT presentation
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Oropharyngeal tularemia Primary disease is confined to the
throat.
Ingestion of infected meat or water
can result in orpharyngeal or
gastrointestinal tularemia
Oculoglandular tularemia
Inoculation of the conjunctivae
Unilateral, purulent conjunctivitis
preauricular, submandibular or cervical lymphadenopathy
http://images.google.com/imgres?imgurl=http://www.nzma.org.nz/journal/120-1248/2403/content01.jpg&imgrefurl=http://www.nzma.org.nz/journal/120-1248/2403/&usg=__FWs34bcKtueXqBUhsUdoxII3tt8=&h=287&w=348&sz=16&hl=en&start=62&um=1&tbnid=BtCvQ73Uz2jaLM:&tbnh=99&tbnw=120&prev=/images%3Fq%3DOculoglandular%2Btularemia%26ndsp%3D20%26hl%3Den%26rls%3Dcom.microsoft:en-us:IE-SearchBox%26rlz%3D1I7ADBR_en%26sa%3DN%26start%3D60%26um%3D1 -
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Pathogenesis
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Daniel L. Clemens,* Bai-Yu Lee, and Marcus A. Horwitz.
INFECTION AND IMMUNITY, Sept. 2005, p. 58925902
Macrophages engulf F. tularensis within
a pseudopod loop
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Facultative intracellular pathogen
Capsule protects against complement killing
Macrophage uptake
bacterial surface polysaccharides
serum complement
complement C3 receptors
LPS - O antigen
prevents maturation of the phagosome multiply to high levels in cytosol
Bacterial release via apoptosis
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Diagnosis
Symptoms & History
Direct staining of clinical specimens with a
fluorescein-labeled antibodies. Serum antibody titers of 1:160 or greater
Culture on cysteine-rich media
Notify Laboratory personnel if you suspectFrancisellasince it is HIGLY INFECTIOUS
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Treatment of Tularemia
Prompt removal of ticks and insect repellent can
prevent disease.
Antibiotics
Streptomycin is the drug of choice
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Yersinia
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Overview: 3species cause
human disease
Yersinia pestis
Yersinia enterocolytica
Yersinia pseudotuberculosis
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Overview: Plague
Yersinia pestis; a gram-negative bacterium.
Three forms of clinical illness;
Bubonic
Septicemic
Pneumonic
Pneumonic is the only one transmittedthrough aerosals.
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Plague: Overview
Natural disease of rodents
Fleas that live on rodents transmit the
bacteria to humans, in the bubonic form.
This disease occurs in many areas of the
world, including the United States.
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Microscopic Characteristics
Y. pestisappear as
single cells or short
chains of plump,gram-negative rods.
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Microscopic Characteristics
Gram stain:
In direct smears,
bacterial cells may be
inside or outside of
leukocytes.
The Gram smear
morphology is
suggestive but not
specific for Y. pestis.
Bipolar staining of a plague smear
prepared from lymph aspirated
from a bubo of plague patient.
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Microscopic Characteristics
Bipolar staining
occurs when usingWayson, or
Giemsa stain.
CDC
C l Ch i i
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Colony Characteristics
Grows well on most
standard laboratory
media.
Sheep Blood Agar
Gray-white
translucent colonies
Pinpoint, gray-
white, non-hemolytic
at 24 hoursBlood agar plate of Yersinia pestisat 48 hours.
CDC/Dr. Brodsky
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Non-motile
Pleomorphic gram-negative bacillus
Urease, and oxidase negative
Facultative anaerobe
Optimal growth at 28o C
Facultative intracellular parasite
Y. pestis: Physiology
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Public Health Aspects of Plague
Fleas carry Y. pestisin their intestinal tract.
When feeding the fleas regurgitate uncapsulated
organisms.
Bacteria re-encapsulate and grow.
Progeny are resistant to intracellular killing
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Yersinia pestis life-cycle
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Plague - Clinical types
Bubonic
infected lymph nodes.
Pneumonic (most likely BT presentation)
transmissible by aerosol; deadliest.
Septicemic blood-borne organisms.
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Bubonic Plague
Regional lymphadenitis (Buboes)
Inguinal, axillary, or cervical lymph nodes mostcommon
80% can become septic
60% mortality if untreated
Cutaneous findings
Possible papule, vesicle, or pustule at inoculation site
Purpuric lesions - late
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Bubo swollen inguinal lymph node or bubo.
After the incubation period of 2-7 days, symptoms
of the plague appear.
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Pneumonic Plague
Pneumonic
From aerosol or septicemic spread to lungs.
Person-to-person transmission by respiratorydroplet.
100% mortality untreated.
Pneumonia progresses rapidly to dyspnea,cyanosis.
Death from respiratory collapse/sepsis.
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Primary or secondary
Secondary from bubonic or pneumonic forms
100% mortality if untreated
Severe endotoxemia
Systemic inflammatory response syndrome
Shock, Disseminated intravascular coagulopathy(DIC)
Adult Respiratory Distress Syndrome (ARDS)
Septicemic Plague
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This patient presented with symptoms of plague that included gangrene of the right hand causing necrosis of the fingers.
In this case, the presence of systemically disseminated plague bacteria Y. pestis, i.e. septicemia, predisposed this patient to abnormal
coagulation within the blood vessels of his fingers.
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Y. pestis:Virulence Determinants
3 virulence encoded Plasmids
Virulence is up-regulated at 37C
Capsule (F1 antigen)
Yersinia Outer Proteins
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Yersinia Outer Proteins
(Yops)
11 different proteins
Antiphagocytic
Inhibit production
proinflammatory cytokines tumor necrosis factor
Cytotoxin
Y
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Yops Targets
dendritic cells
macrophages
Neutrophils
does not target B and T lymphocytes
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F-1 Antigen
Glycoprotein capsule expressed at 370C
Not expressed in flea host
Antiphagocytic
Antibodies to F-1 are protective
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Plasminogen activator
(fibrinolysin) and Coagulase
Plasmid encoded proteins
Promote dissemination of organisms from the clotat the bite site
Coagulase is produced at 280C but not at 320C.
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Diagnosis
Examination of Bubo
aspirate, blood, sputum
stained for bipolar staining
Fluorescent-antibody
Culture (hazardous)
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Plague Treatment
Y. pestisis susceptible to a variety of
antibiotics.
streptomycin, tetracycline, and doxycycline
Peumonic plague is contageous and
isolation is recommended.
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Clinical Case
30 year old man from Colorado, went to ahospital emergency department with a 3-
day history of fever, nausea, vomiting, and
right inguinal lymphadenopathy.
Patient was not a hunter nor had he been
in the woods recently but he did have
dogs.
He was discharged home without
treatment.
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Three days later, the man returned and
was hospitalized with sepsisand bilateral
pulmonary infiltrates.
One of the patient's dogs had serologicevidence of past Y. pestis infection.
Cultures of blood and a lymph nodeaspirate.