Bart_Borelioza_CNS.pdf

Running title: Bartonella henselae in central nervous system

Bartonella henselae and Borrelia burgdorferi infections of central nervous system

Edyta Podsiadły*, Tomasz Chmielewski, Stanisława Tylewska-Wierzbanowska

National Institute of Hygiene, 24 Chocimska street, 00-791 Warsaw, Poland

Phone: +48225421261, fax: +48228493335, e-mail: epodsiadly@pzh.gov.pl

Key words : mixed infection, central nervous system, B. henselae

To investigate the role of B. henselae in patients with symptoms suggesting neuroborreliosis, serum

and cerebrospinal fluid samples were tested with serological and PCR methods. Among 17

examined patients, in 12 cases B. burgdorferi infections were detected, in 1 case B. henselae

infection was ascertained and in two patients mixed B. burgdorferi and B. henselae infections were

found. These results indicate that mixed infections should be taken into consideration in

establishing diagnosis of neurological disorders. The conclusion needs further studies.

Introduction

The aim of our study was to investigate the role of B. henselae in central nervous system disorders

among patients with recognized neuroborreliosis. Since B. henselae and B. burgdorferi sensu lato

shared the same tick vector – Ixodes sp. mixed infections should be considered in the differential for

patients who present with neurological symptoms such as seen with borreliosis. Lyme disease is the

most frequent tick-borne disease in the world. Neurologic abnormalities are prominent in this

disease. During the first stage lymphocytic meningitis with cranial or peripheral nerve palsy and

radiculoneuritis accompanied by stiff neck and headache may occur. In some cases a wide spectrum

of clinical manifestations including encephalopathy, polyneuropathy or encephalomyelitis in course

of disease are present (1).

In 11% of patients with cat-scratch disease neurological symptoms are observed. They include

encephalitis, cerebral arterititis and radiculitis (2). Mixed infection of the central nervous system by

B. burgdorferi sensu lato and B. henselae were described (3).

Material and methods

Seventeen paired serum and cerebrospinal fluid samples were studied. Materials from patients with

various clinical symptoms suggesting neuroborreliosis were sent from various Polish hospitals.

Among them, 9 patients had symptoms of meningitis (headache, nauseea, vomiting and mild neck

stiffness), two persons - sclerosis multiplex, two ones had a headache. In the remaining persons, in

one case double vision with difficulty walking, in the second mediastinal lymphadenitis with

pulmonary interstinal changes, in third depression with paresis of face muscles, in the last one

bilateral facial nerve palsy were observed.

Specific antibodies to B. burgdorferi sensu lato in serum were tested with recombinant antigens p21

(OspC), p41i (inner part of flagellin) for IgM and p21, p41i, p18, p100 for IgG class in ELISA test

(BIOMEDICA, Austria). Serum samples were also tested for the presence of B. henselae and B.

quintana specific antibodies. Levels of serum IgM and IgG immunoglobulins were measured with

indirect immunofluorescence test (MRL Diagnostic, USA).

DNA was extracted from the cerebro-spinal fluid samples with QIAamp tissue kit (QIAGEN,

Germany) according to manufacturer recommendations. Extracted DNA was subjected to PCR

amplification of the 16S rRNA for B. burgdorferi sensu lato (5) and 16S-23S rRNA fragment gene

characteristic for Bartonella species (4).

Reactions were performed in a final volume of 50 ml containing: 10 mM Tris-HCl, 50 mM KCl, 3,5

mM MgCl 2 , 0.1% gelatin, 200 mM dNTPs, 50 pmol of each primer and 2 U of Taq DNA

polymerase (Perkin-Elmer Cetus, USA). Aliquots of 5 ml of DNA template were added to each

reaction mixture. PCR was run in a Mini Cycler apparatus (MJ Research, USA).

Results

Fourteen of 17 examined patients had IgM or IgG antibodies to B. burgdorferi. Three patients with

clinical symptoms suggesting neuroborreliosis were seronegative. The 16S rRNA B. burgdorferi

gene fragment was not detected in CSF samples.

B. henselae infection was detected in three patients.

Patient No. 1

She suffered from meningitis. DNA of B. henselae was found in CFS, he had antibodies to B.

burgdorferi. Although specific antibodies to B. henselae were not found in his serum, it may be

regarded as a mixed infection.

Patient No. 2

Patient suffered also from meningitis. He had IgG antibodies to B. henselae and IgM antibodies to

B. burgdorferi at borderline level. What might indicate a probable mixed infection.

Patient No. 3

Patient complained of headache as a lasting symptom after meningitis due to neuroborreliosis. Two

months before neuroborreliosis was recognized serologically and ceftriaxone regimen was applied.

At the time of testing he was seronegative to B. burgdorferi , but antibodies to B. henselae were

found in titer 64. It might suggest a single B. henselae infection.

Conclusions

These results confirm that B. henselae can be an etiological agent of a central nervous system

infection with symptoms resembling neuroborreliosis. Therefore B. henselae infection should be

considered on the differential when evaluating patients suspected of having neuroborreliosis. Mixed

infections of central nervous system with B. burgdorferi and B. henselae should be taken into

consideration especially in patient group with incomplete resolution of Lyme borreliosis symptoms

after treatment. The conclusion needs further investigation.

References

1. Steere, A. C. 1989. Medical progress. Lyme disease. N. Engl. J. Med. 321 :586-596.

2. Schwartzman, W. A. 1992. Infections due to Rochalimea : The expanding clinical spectrum.

Clin. Infect. Dis. 15 :893-902.

3. Eskow, E. & R.V. Rao & E. Mordechai. 2001. Concurrent infection of the central nervous

system by Borrelia burgdorferi and Bartonella henselae : evidence for a novel tick-borne disease

complex. Arch. Neurol. 58 :1345-7.

4. Jensen, W. A. & M. Z. Fall & J. Rooney, et al. 2000. Rapid identification and differentiation of

Bartonella species using a single-step PCR assay. J. Clin. Microbiol. 38 :1717-22.

Tylewska-Wierzbanowska, S. & T. Chmielewski. 2001. Improvement of laboratory methods for

Lyme borreliosis recognition. Clin. Microbiol. Infect. 7 suppl.1 :194 (P970).

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