BARTONELLOSIS: ONE HEALTH PERSPECTIVES on an EMERGING INFECTIOUS DISEASE Edward B

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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

BARTONELLA SPECIES

• 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

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

My Thought: Is Bartonellosis A Modern Day “Hidden” Epidemic?

Bubonic Plague 1411

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 1990,s Angiomatosis 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 Chomel B B, et al. Bartonella endocarditis: A pathology shared by animal reservoirs and patients. Ann NY Acad Sc 1166: 120-126, 2009. HUMAN YEAR CANINE YEAR B. quintana 93 B. vinsonii 95 B. elizabethae 93 (berkhoffii) B. henselae 93 B. clarridgeae 01 B. washoensis 98 B. vinsonii arup. 99 B. washoensis 03 B. vinsonii berk. 00 B. quintana 05 B. koehlerae 04 B. henselae 09 B. alsatica 05 B. koehlerae 09 B. mayotimonensis 10 B. rochalimae 09

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

Regurgitation blood meal 1

6 5 2,3

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 Blood BAPGM Bk IFA Bh IFA Bvb IFA Day PCR PCR 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 • Epistaxis • Wt Loss, Seizures, • Endocarditis Vasculitis

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

BARTONELLA SPP. DETECTION AND ISOLATION USING BAPGM

Sample DNA

BAPGM culture DNA 16S-23S PCR

- + Plate Isolates DNA

Sequence

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 Sample Source Direct Enrichment Isolate /Date: FATHER Culture Blood 10/18 Bvb II Neg Neg

Blood 11/5 Neg Bvb II Neg CSF 11/5 Neg Bvb II Oral Swab 12/11* Bvb II NA NA Saliva B spp NA NA Bld ACD 1/18 Neg Bh (H1) Neg Bld EDTA 1/18 Neg Bh (H1) Neg Breitschwerdt EB, et al. Parasites and Vectors, Online, Open Access 2010

OCCULT INFECTION: B. VINSONII SUBSP BERKHOFFII

Sample Source Direct Enrichment Isolate /Date: Daughter Culture Blood 11/07 Neg Bvb II Neg

Blood 11/28* Neg Bvb II Neg Serum 11/28 Neg Bvb II Neg Blood 01/11 Neg Bvb II Bvb II

Blood 03/20 Neg Bvb II Neg Serum 03/20 Neg Neg Neg 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 B. henselae Paralysis 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 Blood BAPGM Bh IFA Bq IFA Bvb IFA Day PCR PCR 0 Bh Neg 256 128 256 (H1) 31 Bh Bh 1024 256 256 *Isolate (H1) (H1)* 256 Neg Bh NT NT NT (CSF) (H1) 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

Day Post- Direct Enrichment Bvb I inoculation Culture Seroconversion 5 Neg Neg <16

33 Bvb I* Neg <16

81 Neg Bvb I 32 (128) 234 Neg Neg <16

*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 Center for Comparative Medicine – Dr. Linda Kidd and Translational Research – Dr. Ashlee Duncan

OUR COMMUNITY: OUR WORLD: ONE HEALTH

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