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BARTONELLOSIS: ONE HEALTH

PERSPECTIVES ON AN EMERGING

INFECTIOUS DISEASE

Edward B. Breitschwerdt, DVM

CENTER FOR COMPARATIVE MEDICINE AND

TRANSLATIONAL RESEARCH

North Carolina State University College of Veterinary

Medicine

&

Galaxy Diagnostics, Research Triangle Park, NC

• Gram negative, short pleomorphic rods • Family: BARTONELLACEAE

● Order: Rhizobiales ● Class: Alpha Proteobacteria

• Fastidious: Division time 22-24 hrs • Cellular targets:

•Erythrocytes •Endothelial Cells •Microglia Cells •Macrophages •CD34 Progenitor Cells

BARTONELLA SPECIES

Kordick DL, Breitschwerdt EB: J Clin

Microbiol. 33: 1655-56, 1999

Rhizobiales

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THE STATE OF BARTONELLOSIS!

“It is the mark of an educated mind to be able

to entertain a thought without accepting it”

Aristotle

Bubonic Plague 1411

My Thought: Is Bartonellosis

A Modern Day “Hidden”

Epidemic?

HOW CAN YOU HIDE AN

EPIDEMIC?

PERHAPS IT IS EASY:

1. Start with an unknown genus of bacteria.

2. Behave as a stealth pathogen that is difficult to detect or isolate.

3. Maintain a large and diverse pet and wildlife reservoir in nature.

4. Facilitate transmission through multiple vectors and by other means.

BARTONELLA, HIV, PCR AND

VASOPROLIFERATIVE DISEASE!

Bacillary

Angiomatosis

1990,s

Peliosis Hepatis

Relman D, et. al.NEJM: 1990

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BARTONELLA SPECIES AND

VASOPROLIFERATIVE DISEASES

• Bacillary Angiomatosis (Dogs & Humans)

• Spontaneous Splenic Rupture (Jaguar & Human)

• Hemangiopericytoma in a 12 year-old Female English Sheepdog*

• Epitheloid Hemangioendothelioma in a 13-year-old male (Hepatic)*

• EHE in patients from Australia and England**

• Hemangiopericytoma (Dog, Wolf, Horse)***

*Breitschwerdt EB, et. al. JCM, 2009 **Mascarelli PE, et. al. JCM, 2011

***Beerlage C, et al. Bartonella vinsonii subsp. berkhoffii and Bartonella

henselae as potential causes of proliferative vascular disease in animals.

Med Microbiol Immunol 201: 319-26, 2012.

BARTONELLA ENDOCARDITIS

HUMAN YEAR

B. quintana 93

B. elizabethae 93

B. henselae 93

B. washoensis 98

B. vinsonii arup. 99

B. vinsonii berk. 00

B. koehlerae 04

B. alsatica 05

B. mayotimonensis 10

CANINE YEAR

B. vinsonii 95

(berkhoffii)

B. clarridgeae 01

B. washoensis 03

B. quintana 05

B. henselae 09

B. koehlerae 09

B. rochalimae 09

Chomel B B, et al. Bartonella endocarditis: A pathology shared by animal

reservoirs and patients. Ann NY Acad Sc 1166: 120-126, 2009.

THE POSTULATE OF COMPARATIVE

INFECTIOUS DISEASE CAUSATION

Vergil is credited with stating: “Happy is he

who knows the causes of things”.

Bartonella spp. and disease causation:

Endocarditis

Granulomatous Inflammatory Disease

Chronic Intravascular Infection

Vasoproliferative Tumors

Breitschwerdt EB, et al: Koch’s Postulates and the Pathogenesis of Comparative

Infectious Disease Causation associated with Bartonella species. J Comparative

Pathology 3: 1-11, 2013

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COMPARATIVE DISEASE FEATURES

OF BARTONELLOSIS

Canine Human

Peliosis hepatis + +

Bacillary angioma + +

Endocarditis + +

Myocarditis + +

Granulomatous

Lymphadenitis + +

Hepatitis + +

Encephalitis + +

ITP/IMHA + +

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. bovis – horn fly, Haematobia spp

Source: www.earlham.edu

Feces

Internalization

1

Regurgitation

2,3 5 6

blood meal

Adapted and modified from Dehio et al. Nature 2005

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BARTONELLA: RESERVOIR

HOSTS

Reservoirs:

• Rodents

• Cats

• Dogs

• Wildlife

• Ruminants

• Human beings

• Others

BARTONELLA MELOPHAGI: NEW

ZOONOTIC BARTONELLA SPP.

Bartonella melophagi DNA Sequencing

Sheep keds (Michael Kosoy)

Sheep Blood (David Bemis)

Isolation from Human Blood (BAPGM Platform)

Patient 1: Pericarditis- California

Patient 2: Chronic fatigue syndrome-

Ohio/NC Maggi R, Breitschwerdt EB. Emerg Infect Dis 2009

FLEAS, CATS AND

BARTONELLA SPECIES • B. henselae

• B. clarridgeiae

• B. koehlerae

• B. bovis?

• B. quintana?

FLEA CONTROL!

Pet/Flea Cycle

Wildlife/Pet/Flea Cycle

Photo: Stephen Jaffee DVM

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BARTONELLA KOEHLERAE AND

NEUROCOGNITIVE ABNORMALITIES

• 18 Year-old Woman

• Peripheral Visual Loss, Sensory Neuropathy,

Hallucinations (4 year history)

• Progression: Headaches, joint pain, back

pain, mood swings, dizziness

• Neurologist, Psychiatrist, Neuro-ophthalmol

• B. koehlerae seroreactive, BAPGM

enrichment blood culture positive Breitschwerdt EB, et al. J Clin Microbiol 49: 3415-17, 2011.

BARTONELLA KOEHLERAE Breitschwerdt EB. et al J Clin Microb, 2011

Sample

Day

Blood

PCR

BAPGM

PCR

Bk IFA Bh IFA Bvb IFA

0 Bspp

Neg 64* <16 <16

294 Neg Bk

256 <16

<16

507 Neg Bk

256 <16

<16

801 Neg Neg

<16

<16

<16

TICK TRANSMISSION OF

BARTONELLA SPECIES Billeter SA, Levy MG, Chomel BB, Breitschwerdt EB:

Vector Transmission of Bartonella species with emphasis on the potential for tick transmission. Med & Vet Entomol 22: 2008.

YES (1995-2005)

Case-based evidence. (Dogs and Humans)

Epidemiological Data (Dogs)

PCR Data (Ticks)

MAYBE (2008)

Vector transmission of Bartonella henselae by Ixodes ricinus. EID, 2008.

YES!- Reis C, et al. Plos Neglected Trop Dis. 2011

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BVB GENOTYPE I:

HISTORICAL PROGRESSION

• Polyarthritis

• Wt Loss, Seizures,

Vasculitis

• Epistaxis

• Endocarditis

BARTONELLA IFA

B. HENSELAE B. VINSONII SUBSP.

BERKHOFFII

WILL AN INSECT TISSUE CULTURE

MEDIUM ENHANCE THE GROWTH

OF FASTIDIOUS BACTERIA?

Christmas Holiday , 2003

Dr. Sushama Sontakke

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GALAXY DIAGNOSTICS

www.galaxydx.com Breitschwerdt EB, Sontakke S. US Patent #

7,115,385 Media and Methods for Cultivation of Microorganisms. Issued October, 2006

Chief Scientific Officer

A company devoted to diagnostic documentation of infections with Bartonella spp. and other fastidious bacteria.

Transfer of BAPGM isolation approach to CDC, Duke University & other researchers

Kosoy M, et al. Bartonella tamiae isolated from patients from Thailand J

Clin Microbiol, 46: 772-775, 2008

16S-23S PCR

Sequence

BAPGM culture

Sample DNA

DNA

Plate Isolates DNA

+ -

BARTONELLA SPP. DETECTION AND

ISOLATION USING BAPGM

Duncan AW, et al: A combined approach for the enhanced

detection and isolation of Bartonella species. J Microbiol

Methods 69:273-81, 2007

BARTONELLA VINSONII SUBSP.

BERKHOFFII & HUMAN INFECTION

Father 52 year-old male veterinarian

Progressive weight loss (HIV/cancer?)

Muscle weakness, Incoordination

Poorly defined vascular lesion MRI

Daughter 12 year-old, previously healthy

Headache, muscle pain, insomnia

Mother- BAPGM blood culture negative

Two Sons- BAPGM blood culture negative

Breitschwerdt EB, et al. Parasites and Vectors, Online, 2010

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CO-INFECTION: B. VINSONII & B.

HENSELAE

Neg

Neg

Bh (H1)

Bh (H1)

Neg

Neg

Bld ACD 1/18

Bld EDTA 1/18

NA

NA

NA

NA

Bvb II

B spp

Oral Swab 12/11*

Saliva

Neg Bvb II

Bvb II

Neg

Neg

Blood 11/5

CSF 11/5

Neg Neg Bvb II Blood 10/18

Isolate

Enrichment

Culture

Direct Sample Source

/Date: FATHER

Breitschwerdt EB, et al. Parasites and Vectors, Online, Open Access 2010

OCCULT INFECTION: B.

VINSONII SUBSP BERKHOFFII

Neg

Neg

Bvb II

Neg

Neg

Neg

Blood 03/20

Serum 03/20

Bvb II

Bvb II

Neg Blood 01/11

Neg

Neg

Bvb II

Bvb II

Neg

Neg

Blood 11/28*

Serum 11/28

Neg Bvb II

Neg Blood 11/07

Isolate

Enrichment

Culture

Direct Sample Source

/Date: Daughter

Breitschwerdt EB, et al. Parasites and Vectors Online, Open Access, 2010

BARTONELLA BACTEREMIA AND

NEUROLOGICAL DYSFUNCTTION

Patient Age Gender Neuro Abnor Barton sp.

1 23 F Seizures B. henselae

2 41 F H/I/ML B. henselae

3 44 F H/I/ML Bh/Bvb

4 54 F H/I/ML B. henselae

5 49 M Partial

Paralysis

B. henselae

6 14 M Migraines B. henselae

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SEIZURES WITH PERSISTENT

BARTONELLA HENSELAE Breitschwerdt EB. et al J Clin Microb 46:2856, 2008

Sample

Day

Blood

PCR

BAPGM

PCR

Bh IFA Bq IFA Bvb IFA

0 Bh

(H1)

Neg 256 128 256

31

*Isolate

Bh

(H1)

Bh

(H1)*

1024 256 256

256

(CSF)

Neg Bh

(H1)

NT NT NT

458

Neg Neg 64 64 64

HUMAN Perinatal TRANSMISSION

OF BARTONELLA SPP.

Father- B. vinsonii subsp. berkhoffii

Mother- B. vinsonii subsp. berkhoffii/B. henselae

Placental tissues (1998)- PCR negative

Cervical biopsy (1991)- B. henselae

Son (10 Yrs-old)- B. vinsonii subsp. berkhoffii/B. henselae

Daughter (Twin, died 8 days of age)- B. vinsonii subsp. berkhoffii/B. henselae

Entire family has been sick for 10-20 years. Breitschwerdt EB, et al J Clin Microbiol 48:2289, 2010.

NEEDLE STICK TRANSMISSION OF

BARTONELLA VINSONII (BERKHOFFII)

• 7 Year-old male Newfoundland Dog

• Referral- multiple cutaneous tumors

• Cytology- Histiocytic, lymphocytic or

undifferentiated epithelial neoplasia

• Fractious Dog- inadvertent needle stick

• Ehrlichiosis- Dx and Tx 5 months earlier

• ??? Needle Stick Transmission

• No seroconversion to Ehrlichia canis, PCR

negative

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NEEDLE STICK TRANSMISSION OF

BARTONELLA TO A VETERINARIAN

<16

Neg

Neg

234

32

(128)

Bvb I

Neg 81

<16 Neg

Bvb I* 33

<16 Neg

Neg 5

Bvb I

Seroconversion

Enrichment

Culture

Direct Day Post-

inoculation

*Headaches, Parathesias

BARTONELLOSIS: HIGH RISK AND

RHEUMATOLOGICAL PATIENTS

PARAMETER HIGH RISK RHEUMATOLOGIC

Patient # 192 296

Male/age 55/51 91/36

Female/age 137/40 205/46

Seroreactive (%) 49 63

PCR + (%) 24 41 Maggi RG, Mascarelli PE, Pultorak EL, Hegarty BC, Bradley JM, Mozayeni BR, Breitschwerdt

EB. Bartonella spp. bacteremia in high-risk immunocompetent patients. Diagn Microbiol

Infect Dis. 2011 Dec;71(4):430-7.

Maggi RG Pultorak EL, Mozayeni BR, Hegarty BC, Bradley JM, Correa, MT, Breitschwerdt

EB. Bartonella henselae, Bartonella koehlerae and Bartonella vinsonii subsp. berkhoffii

bacteremia in rheumatic patients from a Lyme disease endemic region. Emerging Infect Dis

18: 783-791, 2012

BARTONELLOSIS: A PROBLEM

IN ONE HEALTH

ENDOCARDITIS YEAR RESERVOIR VECTOR

B. quintana 93 Human/Cat Louse

B. elizabethae 93 Rat Flea

B. henselae 93 Cat Flea

B. washoensis 98 Gr. Squirrel Flea?

B. vinsonii arup. 99 Mice Tick?

B. vinsonii berk. 00 Dog/Coyote Tick?

B. koehlare 04 Cat Flea

B. alsatica 05 Rabbit Scratch

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ACKNOWLEDGEMENTS

• Colleagues

• VBDDL Staff

• Internal Medicine Residents

• Barbara Hegarty

• Ph.D. Graduate Students – Dr. Dorsey Kordick

– Dr. Brandee Pappalardo

– Dr. Jiraporn Suksawat

– Dr. Adam Birkenheuer

– Dr. Linda Kidd

– Dr. Ashlee Duncan

Center for Comparative Medicine

and Translational Research

OUR COMMUNITY: OUR WORLD: ONE HEALTH

Questions??