Breitschwerdt

BARTONELLA SPECIES:TICK TRANSMISSION AND CHRONIC INTRAVASCULAR INFECTION : 

BARTONELLA SPECIES:TICK TRANSMISSION AND CHRONIC INTRAVASCULAR INFECTION Edward B. Breitschwerdt DVM Intracellular Pathogens Research Laboratory College of Veterinary Medicine, North Carolina State University, Raleigh, NC,

BARTONELLA: GRAM NEGATIVE ALPHA PROTEOBACTERIA: 

BARTONELLA: GRAM NEGATIVE ALPHA PROTEOBACTERIA Short, Pleomorphic rods Fastidious Argryophilic Hemotropic Endotheliotropic Dendritic Cells Vector-borne Scratches Bites

COMPARATIVE DISEASE FEATURES OF BARTONELLOSIS: 

COMPARATIVE DISEASE FEATURES OF BARTONELLOSIS Dog Human Peliosis hepatis + + Endocarditis + + Myocarditis + + Granulomatous Lymphadenitis + + Hepatitis + + Encephalitis + + ITP/IMHA + +

A PROBLEM IN COMPARATIVE MEDICINE: 

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?

KNOWN VECTOR TRANSMITTED BARTONELLA SPP.: 

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

TICK TRANSMISSION OF BARTONELLA SPECIES: 

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.

CLINICAL CASES IN HUMAN MEDICINE: 

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

BARTONELLA AND TICK TRANSMISSION: 

BARTONELLA AND TICK TRANSMISSION January Tick Attachment Granulomatous Lymphadenitis Argryophilic Bacteria Seroreactivity PCR+ Pappalardo BL, et. al. J Vet Intern Med, 2000

SNEEZY: 

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)

SNEEZY: 

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 +)

B. VINSONII BERKHOFFII SEROEPIDEMIOLOGY AND TICK TRANSMISSION: 

B. VINSONII BERKHOFFII SEROEPIDEMIOLOGY AND TICK TRANSMISSION

MOLECULAR EPIDEMIOLOGICAL SURVEYS WITHIN THE UNITED STATES: 

MOLECULAR EPIDEMIOLOGICAL SURVEYS WITHIN THE UNITED STATES

MOLECULAR EPIDEMIOLOGICAL SURVEYS OUTSIDE THE UNITED STATES: 

MOLECULAR EPIDEMIOLOGICAL SURVEYS OUTSIDE THE UNITED STATES

BARTONELLA SPECIES INDUCE PERSISTENT BLOOD-BORNE INFECTIONS: 

BARTONELLA SPECIES INDUCE PERSISTENT BLOOD-BORNE INFECTIONS YES! Bartonella bacilliformis- Verruga peruana Homeless Individuals- B. quintana- 78 weeks HIV-infected Individuals San Francisco (Fever) n=382, 18% South Africa (Outpatients) n=188, 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

HISTORICAL HIGHLIGHTS : 

HISTORICAL HIGHLIGHTS PERUVIAN ANDES B. BACILLIFORMIS Source: www.andesmountain.cl

HISTORICAL HIGHLIGHTS: 

Source: www.ypres-battlefield-tours.com Source: www.eurekalert.org World War I HISTORICAL HIGHLIGHTS

Slide17: 

REAL-TIME PCR OF BARTONELLA HENSELAE: ITS REGION Protocol Data Results Standardization Curve Sample Curve Negative Control (Bld) Positive Control (0.01 pg/ul Bld) Maggi RG, Breitschwerdt EB, J Clin Microbiol 43:1171, 2005

Slide18: 

Bartonella henselae (Houston I) Bartonella quintana (Fuller Strain) BARTONELLA GROWTH IN BAPGM LIQUID MEDIUM* Maggi RG, AW Duncan and EB Breitschwerdt. Novel chemically modified liquid medium that will support the growth seven Bartonella species. J. Clin. Microbiol. 43:2651-55, 2005

Slide19: 

Real-time PCR Conventional PCR Sequence BAPGM culture Sample DNA DNA Plate Isolates DNA + - - + BARTONELLA SPP DETECTION AND ISOLATION USING BAPGM

RIDING INSTRUCTORBreitschwerdt et al. J Clin Microbiol 45:270, 2007): 

RIDING INSTRUCTOR Breitschwerdt et al. J Clin Microbiol 45:270, 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

RIDING INSTRUCTOR: 

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

RIDING INSTRUCTOR: 

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.

BARTONELLA SPECIES AS STEATH PATHOGENS: 

Source: www.airforce.com BARTONELLA SPECIES AS STEATH PATHOGENS Merrell DS, Falkow S: Nature 430:250, 2004

STUDY DESIGN: 

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

DEMOGRAPHIC DATA: 

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 >10 years All reported animal bites or scratches All reported frequent arthropod exposure

MEDICAL HISTORIES: 

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

MICROBIOLOGICAL RESULTS: 

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.

BAPGM HUMAN ISOLATION RESULTS (n=14): 

BAPGM HUMAN ISOLATION RESULTS (n=14)

VETERINARY RESEARCHER: 

VETERINARY RESEARCHER

VETERINARY ASSISTANT: 

VETERINARY ASSISTANT

WILDLIFE BIOLOGIST (52-Yr-old male): 

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

CONCLUSIONS: 

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.

A PROBLEM IN COMPARATIVE MEDICINE: 

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 ?

ACKNOWLEDGEMENTS: 

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