Serum levels of copper, selenium and manganese in forestry workers testing IgG positive for Brucella, Borrelia, and Rickettsia.

The aim of this study is to measure the alterations in the trace levels of serum copper (Cu), selenium (Se), and manganese (Mn) in forestry workers testing immunoglobulin G (IgG)-positive for Brucella, Borrelia, and Rickettsia. The study was conducted on a sample of 758 subjects (560 male and 198 female). All the subjects underwent medical examinations, which investigated particularly the presence of clinical signs compatible with zoonoses, and routine blood tests from venous blood sample, which tested previous immunisation versus cited microorganisms and serum concentration of Cu, Se, and Mn. The subjects were divided according to IgG positivity versus the cited microorganisms. The group of subjects with IgG positive versus Brucella showed statistically significant higher Cu levels than controls, while the Mn levels were not; the group of subjects with IgG positive versus Rickettsia showed higher levels of all three tested metals. The concentration of the examined metals did not show statistically significant difference between IgG-positive subjects versus subjects with Borrelia compared to controls. These data could confirm the role of both Cu and Se  in the regulation of immune response.

Borrelia miyamotoi infections among wild rodents show age and month independence and correlation with Ixodes persulcatus larval attachment in Hokkaido, Japan.

To clarify how Borrelia miyamotoi is maintained in the environment in Hokkaido, we examined Ixodes persulcatus for its prevalence among wild rodents and its tick vector by detecting a portion of the borrelial flaB gene in rodent urinary bladder and blood samples, and from whole ticks. We compared B. miyamotoi infection rates to Borrelia garinii and Borrelia afzelii, which are human Lyme disease pathogens also carried by wild rodents, and which are transmitted by the same vector tick. Whereas B. garinii and B. afzelii showed age dependence of infection rates among wild rodents (18.4% and 9.9% among adults and 6.0% and 3.4% among sub-adults, respectively) when looking at urinary bladder samples, B. miyamotoi infection rates were not age dependent for either blood (4.2% among adults, and 7.9% among sub-adults) or urinary bladder samples (1.0% among adults, and 1.7% among sub-adults). Moreover, while B. garinii and B. afzelii infection rates showed increases across months (June, July [p<0.05] and August [p<0.01] had higher rates than in May for adult rodents with B. garinii, and July and August had higher rates than in May [p<0.01] for adult rodents with B. afzelii), B. miyamotoi infection rates did not show significant month dependence. These differences in month and age dependence led us to suspect that B. miyamotoi may not develop persistent infections in wild rodents, as B. garinii and B. afzelii are thought to. Furthermore, we examined the extent of rodent exposure to I. persulcatus nymphs and larvae throughout most of the tick's active season (May through September), and determined that B. miyamotoi infection rates in sub-adult rodents were correlated with larval burden (p<0.01), suggesting that larvae may be very important in transmission of B. miyamotoi to wild rodents.

N-terminal disulfide-bridging of Borrelia outer surface protein C increases its diagnostic and vaccine potentials.

OspC is the main target for IgM in early-stage Lyme disease. As such it is employed as its native or recombinant form in routine immunoassays for the determination of Borrelia-specific antibodies. However, recombinant OspC has so far not shown the antigenicity of the native protein. The latter contains an intrinsic signal sequence and an adjacent cysteine residue, the attachment site of the lipid membrane anchor which has been discussed to have an adjuvant effect on the immune reaction. In expression experiments, we have found a recombinant variant, an OspC covalently homodimerized via an N-terminal disulfide bridge, that shows a remarkably enhanced antigenicity without lipid attachment. Three such OspCs derived from different Borrelia strains were subsequently expressed in E. coli and purified under non-reducing conditions. In non-reducing SDS-PAGE, OspC(Δ1-18) exhibited a 48-kDa band of dimeric OspC. When incubated with IgM-OspC-positive human sera, the reaction at 48kDa was always stronger than at 24kDa of monomeric OspC(Δ1-18, C19G). A lineblot with OspC(Δ1-18) also showed a higher diagnostic accuracy than that obtained with OspC(Δ1-18, C19G) based on a higher affinity of IgM for the dimeric form. When used for the immunization of mice, dimeric OspC(Δ1-18) induced consistent high-titre antibodies against OspC, whereas OspC(Δ1-18, C19G) failed to provoke significant titres in some animals. We conclude that the disulfide-bridging of 2 OspC molecules via their N termini forms a complex that is more suitable for the determination of IgM-OspC and is a promising candidate for a monovalent vaccine.

Bilateral facial nerve palsy in the course of neuroborreliosis in children-dynamics, laboratory tests and treatment.

PURPOSE: Presentation of four patients with bilateral peripheral facial nerve palsy as a clinical manifestation of neuroborreliosis in children--diagnostic, treatment and prognosis. MATERIAL AND METHODS: In 2002-2004 in The Chair and Department of Developmental Neurology, 24 children from the Wielkopolska region were admitted with diagnosis of borreliosis. Among all the children with borreliosis, confirmed by serologic examination, 4 (16.7%) demonstrated bilateral peripheral facial palsy (PFP). We investigated the presence of IgM class and IgG class specific antibodies in the sera and cerebrospinal fluid (CSF) of 4 patients with bilateral PFP. (Detected by immunoenzymatic methods--ELISA.) RESULTS: Before the occurrence of PFP all the children manifested unspecified systemic symptoms such as headaches, muscle and articulation pains, weakness and in two cases a mood depression. At first all patients demonstrated elevated IgM antibodies and proper levels of IgG antibodies. Control tests administered within 2-14 months later reduction of antibodies was indicated. Two patients demonstrated significant pleocytosis in CSF test, (without the meningeal symptoms). All children were treated with physiotherapeutic procedures and were administered antibiotic intravenously. CONCLUSIONS: PFP is one of the most frequent neurological symptoms of borreliosis in children. In case of acute PFP and especially the bilateral form of PFP, neuroborreliosis is the most probable diagnosis. All children reported PFP at one side first and after several weeks the paresis of the facial nerve on the opposite side usually appeared. The clinical state of children started to improve after the introduction of physiotherapy and this process usually lasted several months.

[A case of Lyme disease with the triad of neurologic manifestations (meningitis, radiculoneuritis, facial nerve palsy) and dermatitis of the nail roots].

A 23-year-old man experienced dysesthesia of the distal parts of four limbs and spilling of drinking water from the right corner of his mouth. He also experienced low grade fever, headache, and nausea. Neurological examination showed weakness of the right frontal, orbicularis oculi and orbicularis oris muscles, atrophy and weakness of the interosseous muscles in both upper limbs and dysesthesia in the distal parts of four limbs. Laségue's sign was also positive on the left side. Examination of CSF showed lymphocytic pleocytosis and an increase of total protein. Serum antibody against Borrelia garinii was positive as determined by ELISA. The patient was diagnosed as a case of Lyme disease. His symptoms were not alleviated by administration of ceftriaxone (2 g/day) for 15 days, and dermatitis of nail roots appeared. Following administration of penicillin G at a high dose (12 million units/day) for 20 days, both neurologic symptoms and dermatitis were alleviated. Since B. garinii was reported to move preferentially toward the sites of low temperature, dermatitis of nail roots may be one of the characteristic features of Lyme disease caused by B. garinii.

Evidence for spirochetal origin of circumscribed scleroderma (morphea).

Acrodermatitis chronica atrophicans (ACA) and morphea are clinically distinct skin diseases with some common features and possible coexistence. We found antibodies to Borrelia burgdorferi in eight of fifteen patients with morphea. Six of them had IgG antibodies and two both IgG and IgM antibodies. Four of the eight seropositive and five of the seven seronegative patients had been treated with high dose penicillin previously. Spirochetal organisms could be cultured in Barbour-Stoenner-Kelly's medium from a skin biopsy of one seropositive untreated patient. Spirochetes were recovered from histological sections in three of eight, two seropositive and one seronegative morphea and in one of three erythema chronicum migrans patients by an avidin-biotin immunoperoxidase method. The similar clinical picture of ACA and morphea, the response to penicillin therapy in both entities, the presence of antispirochetal antibodies, the isolation of spirochetes in culture and the detection of spirochetal organisms on histological sections suggest a close relationship among these diseases. We conclude that morphea may represent a Borrelia infection. The correlation to ACA is discussed.

Tick-borne Borrelia infection in Sweden.

Spirochetes were cultivated from 17% of 114 Ixodes ricinus ticks in the Stockholm area. Three strains of these spirochetes were selected for studies by electron microscopy. These three strains had definite morphological similarities to spirochetes of the genus Borrelia, as judged by the number of flagella, absence of cytoplasmic tubules, and dimensions. The three strains were not identical, but seemed to consist of two different kinds of cells, one with eight and one with eleven flagella. The three strains were also shown to react with a monoclonal antibody that reacts with Lyme disease spirochetes (Borrelia burgdorferi), but not with strains of other Borreliae, Treponemes, or Leptospiras. These results indicate the possibility of transmission of Borrelia spirochetes from ticks to humans in Sweden. The antibody response to one of the spirochetal strains isolated from Swedish I. ricinus was studied in 37 patients with the typical clinical picture of erythema chronicum migrans (ECM), in 45 patients with chronic meningitis (CMe) cured by high-dose intravenous penicillin, in 298 patients with post-infectious arthritis, and in controls. The antibody response was estimated by indirect immunofluorescence assay (IFA) enzyme-linked immunosorbent assay (ELISA). The antibody levels differed significantly between patients with CMe and healthy individuals (p less than 0.001), both with respect to serum antibody levels and CSF-antibody levels. The antibody levels also differed significantly between patients with ECM and healthy controls as measured by ELISA (p less than 0.05), whereas the difference was not significant as measured by IFA. Five of 298 patients with post-infectious arthritis had higher titers than any of the controls, and two of these five patients had titers higher than any patient with CMe or ECM. These results indicate spirochetal aetiology of ECM, and in some patents with CMe or postinfectious arthritis. As a diagnostic test for ECM, both IFA and ELISA were of limited value, since only 5/37 (14%) ECM patients were positive by IFA, and 14/37 (38%) by ELISA. Regarding patients with CMe, 23/45 (51%) were seropositive by IFA and 30/45 (67%) by ELISA. However, measurement of CSF-antibodies were found to be a more sensitive method than measurement of serum antibodies both by IFA and ELISA, since 38/45 (84%) CMe patients were positive by IFA, and 41/45 (91%) by ELISA. In addition, estimation of CSF antibodies was also found to be a more specific method than estimation of serum antibodies.(ABSTRACT TRUNCATED AT 400 WORDS)

Culture of Ethiopian strains of Borrelia recurrentis.

A number of standard bacteriological media with supplements were tested for their ability to support in vitro growth of Ethiopian strains of Borrelia recurrentis. Propagation of 18 out of 21 strains occurred in Trypticase soy yeast broth to which bovine albumin (fraction V), N-acetyl glucosamine, and sodium pyruvate had been added. This medium supported a population of 10(7) organisms per ml and yielded a harvest of four to five times the original inoculum during the logarithmic phase of growth. Maximal yield varied from 1.4 x 10(7) to 3.4 x 10(7) organisms per ml. Generation time in optimal media was 11.3 h. Lesser multiplication of organisms occurred in other media tested. Strains from primary cultures were infective for the green monkey. Recovery of viable organisms from subculture has not been successful.