BARTONELLA SPECIES:TICK TRANSMISSION AND CHRONIC INTRAVASCULAR INFECTION

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BARTONELLA SPECIESTICK TRANSMISSION AND CHRONIC
INTRAVASCULAR INFECTION

• Edward B. Breitschwerdt DVM
• Intracellular Pathogens Research Laboratory
• College of Veterinary Medicine, North Carolina
  State University, Raleigh, NC,

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BARTONELLA GRAM NEGATIVE ALPHA PROTEOBACTERIA

• Short, Pleomorphic rods
• Fastidious
• Argryophilic
• Hemotropic
• Endotheliotropic
• Dendritic Cells
• Vector-borne
• Scratches
• Bites

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COMPARATIVE DISEASE FEATURES OF BARTONELLOSIS

• Dog Human
• Peliosis hepatis
• Endocarditis
• Myocarditis
• Granulomatous
• Lymphadenitis
• Hepatitis
• Encephalitis
• ITP/IMHA

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A PROBLEM IN COMPARATIVE MEDICINE

• ENDOCARDITIS YEAR RESERVOIR VECTOR
• B. quintana 93 Human Louse
• B. elizabethae 93 Rat Flea
• B. henselae 93 Feline Flea
• B. washoensis 98 Gr. Squirrel Flea?
• B. vinsonii arup. 99 Mice Tick?
• B. vinsonii berk. 00 Canine Tick?
• B. koehlare 04 Cat Flea
• B. alsatica 05 Rabbit
  Scratch?

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KNOWN VECTOR TRANSMITTED BARTONELLA SPP.

• B. bacilliformis - sandfly, Lutzomia verrucarum
• B. quintana - human body louse, Pediculus humanus
  humanus
• B. henselae - cat flea, Ctenocephalides felis
• B. grahamii - rodent flea, Ctenphthalmus nobilis
  
• B. taylorii - rodent flea, C. nobilis

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TICK TRANSMISSION OF BARTONELLA SPECIES

• YES!
• Case-based evidence. (Dogs and Humans)
• NO!
• No vector transmission studies.
• MAYBE??
• PCR amplification from ticks.
• Seroepidemiological associations with other
  tick-borne pathogens.

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CLINICAL CASES IN HUMAN MEDICINE

• 1st Association (Lucey et. al. Clin Infect Dis.
  1992)
• Two male patients
• Bartonella henselae cultured from blood
• (fever myalgia, arthralgia, headache)
• Within weeks after tick attachment
• Co-infection Borrelia burgdorferi and B. henselae
• Four patients from New Jersey
• B. henselae amplified from blood
• 2/4 Tick Bite History, 3/4 No Cat Contact
• B. henselae and B. quintana DNA- was amplified
  blood samples from Russian individuals bitten by
  ticks

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BARTONELLA AND TICK TRANSMISSION

• January
• Tick Attachment
• Granulomatous
• Lymphadenitis
• Argryophilic
• Bacteria
• Seroreactivity
• PCR
• Pappalardo BL, et. al. J Vet Intern Med, 2000

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SNEEZY

• 4 yr F Greyhound Trip to Disney World
• 1/16 Removed Engorged Tick Lt. Ear.
• 1/23 Fever, 40 degrees C
• Submandibular Swelling (Amoxicillin,
  Enrofloxacin, Dexamethasone,)
• 1/31 Referral 3 cm Cervical Mass
• Enlarged Prescapular LN
• (Edema w/ Granulomatous Rx)

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SNEEZY

• Aspiration Cytology Spindle Cell Tumor, Giant
  Cell Tumor
• Biopsy Granulomatous Lymphadenitis,
  Wartharin-Stary oval to rod-like bacteria
• B. vinsonii berkhoffii titer 128 (PCR )

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B. VINSONII BERKHOFFII SEROEPIDEMIOLOGY AND TICK
TRANSMISSION
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MOLECULAR EPIDEMIOLOGICAL SURVEYS WITHIN THE
UNITED STATES
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MOLECULAR EPIDEMIOLOGICAL SURVEYS OUTSIDE THE
UNITED STATES
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BARTONELLA SPECIES INDUCE PERSISTENT BLOOD-BORNE
INFECTIONS

• YES!
• Bartonella bacilliformis- Verruga peruana
• Homeless Individuals- B. quintana- 78 weeks
• HIV-infected Individuals
• San Francisco (Fever) n382, 18
• South Africa (Outpatients) n188, 10
• Cat Scratch Disease- B. henselae PCR at
• 3 and 4 months post-CSD (Arvand M, et. al. JCM
  2006)
• Yes?
• Immunocompetent Individuals with Occupational
  and Arthropod Exposure

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HISTORICAL HIGHLIGHTS
PERUVIAN ANDES B. BACILLIFORMIS
Source www.andesmountain.cl
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HISTORICAL HIGHLIGHTS
World War I
Source www.eurekalert.org
Source www.ypres-battlefield-tours.com
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REAL-TIME PCR OF BARTONELLA HENSELAE ITS REGION
Standardization Curve
Protocol
Data Results
Sample Curve
Negative Control (Bld) Positive Control (0.01
pg/ul Bld)
Maggi RG, Breitschwerdt EB, J Clin Microbiol
431171, 2005
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BARTONELLA GROWTH IN BAPGM LIQUID MEDIUM
Bartonella henselae (Houston I)
Bartonella quintana (Fuller Strain)
Maggi RG, AW Duncan and EB Breitschwerdt. Novel
chemically modified liquid medium that will
support the growth seven Bartonella species. J.
Clin. Microbiol. 432651-55, 2005
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BARTONELLA SPP DETECTION AND ISOLATION USING BAPGM
Sample
DNA
Real-time PCR
BAPGM culture
DNA

-
Plate Isolates
DNA
Conventional PCR
Sequence
-

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RIDING INSTRUCTORBreitschwerdt et al. J Clin
Microbiol 45270, 2007)

• Oct. 2, 2004 (42 Year-old female)
• Bite Lt Hand by Feral Barn Cat
• Cellulitis with Surgical Debridement
• Amoxicillin clavulante 48 hrs
• Ceftriaxone sodium 10 days
• Oct. 23, 2004
• Bite Lt Wrist by 8 Yr-M-German Shepherd
• Dog Hit by truck and painful
• Infection Amoxi-7 days, Levofloxacin-7days

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RIDING INSTRUCTOR

• November 22, 2004
• Due to bite episodes BAPGM Culture
• Isolate Bartonella quintana
• December 16, 2004
• Serology Bq 128, Bvb 256, Bh 64
• Dog Culture Neg/Seronegative
• January 5, 2005
• Feral cats adopted by riding instructor
• Serology Bartonella henselae 4096 1024

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RIDING INSTRUCTOR

• Cat and Human B. quintana isolates were identical
  based upon 16S-23S ITS region.
• Cat and Human sequences differed by one base from
  B. quintana (Fuller, 523/524 bases).
• Pap 31 bacteriophage associated gene could not be
  amplified from cat or human isolates.
• Jan 31, 2005 BAPGM/PCR- Neg.
• May 24, 2005 BAPGM/PCR- Neg.
• No systemic Illness in a 2 year follow-up period.

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BARTONELLA SPECIES AS STEATH PATHOGENS
Merrell DS, Falkow S Nature 430250, 2004
Source www.airforce.com
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STUDY DESIGN

• Feasibility Study (BAPGM/RT-PCR)
• Convenience Sample
• IRB and Biosafety Approval
• Bartonella species/strain determined by DNA
  sequencing.
• Bartonella serology (IFA) retrospectively
• Questionnaire for epidemiological data

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DEMOGRAPHIC DATA

• Sex 12 females/2males
• Age 30-53 years
• Occupations 5 veterinarians, 5 veterinary
  assistants, 1 veterinary researcher, 1 cattle
  rancher, 2 wildlife biologists
• All reported occupational animal contact for
• gt10 years
• All reported animal bites or scratches
• All reported frequent arthropod exposure

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MEDICAL HISTORIES

• Self assessment 9/14 reported chronic illness, 5
  reported infrequent, recurrent illness.
• Symptoms
• Fatigue 13 Sensation Loss 7
• Joint Pain 10 Balance Problems 7
• Insomnia 9 Headache 7
• Muscle Pain 8 Tremors 6
• Memory Loss 8 Syncope 5

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MICROBIOLOGICAL RESULTS

• OVERALL FINDINGS
• 14 individuals were Bartonella PCR/Culture
• B. henselae and B. vinsonii subsp. berkhoffi were
  the only species detected.
• Isolates 4 B. henselae 1 B. vinsonii
  berkhoffii Type II
• Chronic infection was documented in 7
  participants.
• Co-infection with Bh/Bvb was documented in 4
  individuals.
• 8/14 (57) participants were seroreactive to Bh,
  Bq or Bvb.

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BAPGM HUMAN ISOLATION RESULTS (n14)
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VETERINARY RESEARCHER
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VETERINARY ASSISTANT
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WILDLIFE BIOLOGIST (52-Yr-old male)

• Episodic Illness
• Fever, Chills, Headache,
• Body Ache, Rash
• Episodes
• June, 2001
• September, 2002
• September, 03 (Picture)
• February, 2004 2005
• Rash- Last 3 Episodes
• B vinsonii berkhoffii Type II

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CONCLUSIONS

• Bartonella henselae and Bartonella vinsonii
  subsp. berkhoffii can be detected in blood from
  immunocompetent people.
• Pre-enrichment culture with BAPGM is frequently
  necessary to detect infection.
• Animal bites or arthropod exposure are potential
  sources of transmission.
• Infected individuals may not be seroreactive
• Relationship to chronic or relapsing illness is
  undetermined.

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A PROBLEM IN COMPARATIVE MEDICINE

• ENDOCARDITIS YEAR RESERVOIR VECTOR
• B. quintana 93 Human Louse
• B. elizabethae 93 Rat Flea
• B. henselae 93 Feline Flea
• B. washoensis 98 Gr. Squirrel Flea?
• B. vinsonii arup. 99 Mice Tick?
• B. vinsonii berk. 00 Canine Tick?
• B. koehlare 04 Cat Flea
• B. alsatica 05 Rabbit ?

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ACKNOWLEDGEMENTS

• Collaborators
• VBDDL Staff
• Research Scientists
• Barbara Hegarty
• Ph.D. Graduate Students
• Dr. Dorsey Kordick
• Dr. Brandee Pappalardo
• Dr. Jiraporn Suksawat
• Dr. Adam Birkenheuer
• Dr. Linda Kidd
• Ashlee Duncan
• Sarah Billeter
• Heather Hunt

Bayer Animal Health IDEXX Laboratories