Acrodermatitis chronica atrophicans: clinical and microbiological characteristics of a cohort of 693 Slovenian patients.

BACKGROUND: Information on large groups of patients with acrodermatitis chronica atrophicans (ACA) is limited. METHODS: We assessed clinical and microbiological characteristics of patients with ACA diagnosed at a single medical centre and compared findings in periods 1991-2004 vs. 2005-2018. The cohort is representative of Slovenian ACA patients. RESULTS: We assessed 693 patients: 461 females and 232 males, with median age of 64 years. Median duration of ACA before diagnosis was 12 months. In all but 2 patients, the skin lesions were located on extremities, more often on the lower (70.0%) than the upper (45.2%), bilaterally in 42.4%. Reddish-blue discoloration, swelling, thinning and wrinkling of skin were present in 95.2%, 28.1%, 46.4% and 20.5% of patients, respectively. Overall, 64.4% of patients reported constitutional symptoms, 23.1% had local symptoms, and 20.8% had symptoms/signs of peripheral neuropathy. Nodules, arthritis, joint deformity, muscle atrophy and paresis were rare (<3%). Borreliae were isolated from 200/664 (30.1%) skin samples; 92.8% were Borrelia afzelii. B. garinii and B. burgdorferi s.s. were more often isolated from the skin of male patients (OR = 4.17) and from those with arthropathy (OR = 11.74). Patients included in the more recent period were older, complained less often of constitutional symptoms but more often of local symptoms, and more often had local swelling but less often skin atrophy and bilateral involvement, probably as a consequence of earlier diagnosis. CONCLUSIONS: ACA, typically caused by B. afzelii, usually affects older women. Clinical presentation depends on the duration of illness and probably on the Borrelia species causing the disease.

Tick-borne encephalitis in patients vaccinated against this disease.

BACKGROUND: Information on tick-borne encephalitis (TBE) in patients already vaccinated against the disease is limited. OBJECTIVES: To compare the course and outcome in patients with vaccination breakthrough TBE with findings in patients who developed TBE without previous vaccination. METHODS: All adult patients diagnosed with TBE at a single medical centre during a 16-year period and who had received at least two doses of TBE vaccine before the onset of illness qualified for the study. For each patient with breakthrough TBE, two unvaccinated sex- and age-matched patients, diagnosed with TBE in the same year, were included for comparison. RESULTS: Amongst 2332 patients diagnosed with TBE in the period 2000-2015, 39 (1.7%) had been vaccinated against the disease. Their median age was 59 (20-83) years; 22 of 39 (56.4%) were male. In comparison with unvaccinated patients with TBE, those with breakthrough disease more often experienced a monophasic course of illness (P = 0.006), had a higher CSF leucocyte count (P = 0.005), more often had urine retention (P = 0.012), more often needed ICU treatment (P = 0.009), were hospitalized for longer (P = 0.002) and had more severe acute illness (P = 0.004 for simple clinical assessment, P = 0.001 for severity score). CONCLUSION: In addition to several findings corroborating previous results in patients with vaccination breakthrough TBE, such as older age and the presence of a particular specific serum antibody pattern indicating anamnestic response, findings in this study indicate that the acute illness in patients with breakthrough TBE is more severe than in unvaccinated sex- and age-matched patients who develop the disease.

EAN consensus review on prevention, diagnosis and management of tick-borne encephalitis.

BACKGROUND AND PURPOSE: Tick-borne encephalitis (TBE) is an infection of the central nervous system (CNS) caused by tick-borne encephalitis virus (TBEV) and transmitted by ticks, with a variety of clinical manifestations. The incidence of TBE in Europe is increasing due to an extended season of the infection and the enlargement of endemic areas. Our objectives are to provide recommendations on the prevention, diagnosis and management of TBE, based on evidence or consensus decisions. METHODS: For systematic evaluation, the literature was searched from 1970 to 2015 (including early online publications of 2016), and recommendations were based on evidence or consensus decisions of the Task Force when evidence-based data were not available. RECOMMENDATIONS: Vaccination against TBE is recommended for all age groups above 1 year in highly endemic areas (≥5 cases/100 000/year), but also for individuals at risk in areas with a lower incidence. Travellers to endemic areas should be vaccinated if their visits will include extensive outdoor activities. Post-exposure prophylaxis after a tick bite is not recommended. A case of TBE is defined by the presence of clinical signs of meningitis, meningoencephalitis or meningoencephalomyelitis with cerebrospinal fluid (CSF) pleocytosis (>5 × 106 cells/l) and the presence of specific TBEV serum immunoglobulin M (IgM) and IgG antibodies, CSF IgM antibodies or TBEV IgG seroconversion. TBEV-specific polymerase chain reaction in blood is diagnostic in the first viremic phase but it is not sensitive in the second phase of TBE with clinical manifestations of CNS inflammation. Lumbar puncture should be performed in all patients with suspected CNS infection unless there are contraindications. Imaging of the brain and spinal cord has a low sensitivity and a low specificity, but it is useful for differential diagnosis. No effective antiviral or immunomodulating therapy is available for TBE; therefore the treatment is symptomatic. Patients with a potentially life threatening meningoencephalitis or meningoencephalomyelitis should be admitted to an intensive care unit. In the case of brain oedema, analgosedation should be deepened; osmotherapy and corticosteroids are not routinely recommended. If intracranial pressure is increased, therapeutic hypothermia or decompressive craniectomy might be considered. Seizures should be treated as any other symptomatic epileptic seizures. CONCLUSIONS: Tick-borne encephalitis is a viral CNS infection that may result in long-term neurological sequelae. Since its incidence in Europe is increasing due to broadening of endemic areas and prolongation of the tick activity season, the health burden of TBE is enlarging. There is no effective antiviral treatment for TBE, but the disease may be effectively prevented by vaccination.

Oseltamivir prophylaxis in controlling influenza outbreak in nursing homes: a comparison between three different approaches.

PURPOSE: To assess influenza outbreaks in nursing homes (NHs) using different pharmacological preventive measures. METHODS: We compared characteristics of influenza A outbreaks that occurred during 2011/2012 influenza season in three NHs of similar size (208, 167, and 164 residents in NH1, NH2, and NH3, respectively) implementing comparable treatment approaches and non-pharmacological outbreak control measures but different prophylactic pharmacological interventions including oseltamivir 75 mg o.d. for 10 days for all residents (NH1), for directly exposed residents (NH2), and no prophylaxis (NH3). RESULTS: The proportions of residents who developed acute respiratory infection (ARI) in the course of influenza outbreak were 55/208 (26.4 %) in NH1, 64/167 (38.3 %) in NH2, and 31/164 (18.9 %) in NH3; hospital admission was required in 2/55 (3.6 %), 5/64 (7.8 %), and 5/31 (16.1 %) residents of NH1, NH2, and NH3, respectively, while 1/55 (1.8 %), 1/64 (1.6 %), and 3/31 (9.7 %) residents of the corresponding NHs died during influenza outbreak. The duration of the outbreak was shorter in the NH1 where oseltamivir prophylaxis was instituted for all residents (8 days), than in NHs where selective prophylaxis with oseltamivir and no prophylaxis were used (14 and 12 days, respectively). The calculated vaccine effectiveness in residents was 48, 71, and 44 % in NH1, NH2, and NH3, respectively. Staff members had similar ARI attack rate but in comparison to residents were less often vaccinated against influenza and demonstrated higher influenza vaccine effectiveness. CONCLUSIONS: Comparison of influenza outbreaks in three NHs revealed that the duration of the outbreak was the shortest in the NH where prophylaxis with oseltamivir was given to all residents.

Bacterial infection in elderly nursing home and community-based patients: a prospective cohort study.

PURPOSE: The aim of the study was to assess differences between elderly nursing home (NH) residents and elderly patients living in the community admitted to hospital for bacterial infection. METHODS: We conducted a prospective cohort study with manual data collection. Patients aged 65 years and older from NHs and from the community admitted to hospital for bacterial infection were included in the study. Patient characteristics, site of infection, microorganism profile, and final outcome were assessed and compared between these two subgroups of patients. RESULTS: Two hundred and sixty-two patients were from NHs and 707 were from the community. Patients from NHs were older, had more underlying chronic illnesses, and more often showed characteristics indicating advanced debility. In both groups, the most common site of infection was the respiratory tract. In comparison to the community group, patients from NHs had a higher fatality rate (23.3 vs. 10.9%) and more often experienced functional decline at discharge (27.5 vs. 16.8%). After adjustment for age, sex, presence of comorbidity, and debility, NH residence remained associated with increased in-hospital mortality but not with functional decline. CONCLUSIONS: Elderly patients from NHs admitted to hospital for bacterial infection are older and more debilitated than their counterparts from the community. Microorganisms found in the NH residents are somewhat different from those in the community dwellers. The community dwellers had a better survival rate than those admitted from the NHs after adjusting for age, sex, presence of any comorbidity, and debility.

Respiratory viruses in patients and employees in an intensive care unit.

PURPOSE: To evaluate the frequency of respiratory viruses in a nonselected population of intensive care unit patients and employees and to investigate the clinical as well as the epidemiological association with virological findings. METHODS: Between 12 January and 5 March 2009, nasopharyngeal swabs were collected from 55 intensive care unit (ICU) patients and 41 medical personnel at 16 different time-points and tested for 11 respiratory viruses by single real-time PCR using TaqMan or MGB probes. RESULTS: Among the 55 ICU patients tested, there were 30 virus-positive respiratory specimens (30/173, 17.3%) and 23 patients who tested positive at least once for respiratory viruses (23/55, 41.8%). Only the time from admission to the ICU was associated with the probability of testing positive, with the probability of testing positive decreasing with increasing length of stay (P < 0.001). Of the 418 respiratory specimens collected from the healthcare personnel, 27 (6.5%) tested positive. Seventeen employees tested positive at least once for respiratory viruses (17/41, 41.5%). Among the employees, calendar time (P = 0.03) and having sick contacts at home (P = 0.006) were significantly associated with swab positivity. Among the study population, patients had a significantly higher probability of having a positive swab result than employees. The distribution of viruses differed between the two groups. CONCLUSIONS: Our results suggest that when hygiene precautions are adopted, the possibility of transmitting selected respiratory viruses between patients and personnel is limited. They also point to a greater importance of the community over the hospital environment for acquisition of viral respiratory infections by ICU patients and employees.

Isolation of Borrelia burgdorferi sensu lato from blood of adult patients with borrelial lymphocytoma, Lyme neuroborreliosis, Lyme arthritis and acrodermatitis chronica atrophicans.

BACKGROUND: Reports on patients with European Lyme borreliosis in whom borreliae were isolated from the blood are rare and nearly exclusively limited to those with solitary or multiple erythema migrans. Here we report on patients with other manifestations of Lyme borreliosis in whom borreliae were isolated from their blood. PATIENTS AND METHODS: This is a retrospective review of the medical files of patients diagnosed with borrelial lymphocytoma, Lyme neuroborreliosis, Lyme arthritis and acrodermatitis chronica atrophicans at the Department of Infectious Diseases of the UMC Ljubljana, Slovenia, for whom a borrelia blood culture was ordered. The clinical features of patients whose blood culture tested positive for Borrelia burgdorferi sensu lato were reviewed, and the association between the proportion of patients with a positive blood culture and various clinical manifestations was examined. RESULTS: Borrelia burgdorferi sensu lato was isolated from the blood of 1/53 (1.9%) patients with borrelial lymphocytoma, 6/176 (3.4%) patients with Lyme neuroborreliosis, 1/13 (7.7%) patients with Lyme arthritis, and 3/200 (1.5%) patients with acrodermatitis chronica atrophicans. The time interval from the onset of symptoms attributed to Lyme borreliosis and the blood culture ranged from 1 day to >2 years (median 3.5 weeks). At the time of the blood culture, erythema migrans was present in 4/11 (36.4%) borrelia blood culture-positive patients, i.e. in the patient with borrelial lymphocytoma, the patient with Lyme arthritis and the 2/6 patients with Lyme neuroborreliosis. Only two of these 11 (18.2%) patients had fever at the time of the blood culture. CONCLUSIONS: In European patients with Lyme borreliosis, borreliae can be cultured from the blood not only early in the course of the disease but also occasionally later during disease progression.

Cost-effectiveness of a potential anti-tick vaccine with combined protection against Lyme borreliosis and tick-borne encephalitis in Slovenia.

This study assessed cost-effectiveness of a potential anti-tick vaccine that would protect against both Lyme borreliosis (LB) and tick-borne encephalitis (TBE) in a highly endemic setting of Slovenia. A Markov model was developed to estimate cost-effectiveness of a vaccine with potential combined protection against LB and TBE from the societal perspective. The model expressed time in annual cycles, followed a target population through their lifetime, and applied an annual discounting of 3%. A target population entered the model in a susceptible state, with time dependent probabilities to acquire LB/TBE. Disease manifestations were either resolved within one cycle, or a patient developed LB/TBE sequelae. The vaccination consisted of initial immunization and one revaccination. Estimates of LB/TBE direct and indirect costs, and data on natural course of LB/TBE were obtained from Slovenian databases. Effectiveness of the vaccine with potential combined protection against LB/TBE was derived from studies on existing TBE and LB vaccines, while utility estimates were collected from various literature sources. A vaccine with potential combined protection against LB/TBE was predicted to have an incremental cost of €771,300 per 10,000 vaccinated persons, an incremental utility of 17QALYs and a base-case incremental cost-effectiveness ratio (ICER) of 46,061€/QALY. Vaccine cost, effectiveness and discount rates were identified as the most influential model parameters. A wholesale price for a vaccine shot of €9.13 would lead to cost savings followed by health gains for the vaccination strategy. The base-case ICER was below commonly accepted thresholds of cost-effectiveness, indicating that a combined LB/TBE vaccine might be a cost-effective option in Slovenia. With early Health Technology Assessment becoming increasingly important, this analysis still represents a rare example of cost-effectiveness assessment prior to market authorisation. Although obviously in such a situation some key parameters are unknown, our model sets up a tool to analyse pharmacoeconomic criteria that can help development of a cost-effective health technology, in this case a combined tick-borne diseases vaccine.

To test or not to test? Laboratory support for the diagnosis of Lyme borreliosis: a position paper of ESGBOR, the ESCMID study group for Lyme borreliosis.

BACKGROUND: Lyme borreliosis (LB) is a tick-borne infection caused by Borrelia burgdorferi sensu lato. The most frequent clinical manifestations are erythema migrans and Lyme neuroborreliosis. Currently, a large volume of diagnostic testing for LB is reported, whereas the incidence of clinically relevant disease manifestations is low. This indicates overuse of diagnostic testing for LB with implications for patient care and cost-effective health management. AIM: The recommendations provided in this review are intended to support both the clinical diagnosis and initiatives for a more rational use of laboratory testing in patients with clinically suspected LB. SOURCES: This is a narrative review combining various aspects of the clinical and laboratory diagnosis with an educational purpose. The literature search was based on existing systematic reviews, national and international guidelines and supplemented with specific citations. IMPLICATIONS: The main recommendations according to current European case definitions for LB are as follows. Typical erythema migrans should be diagnosed clinically and does not require laboratory testing. The diagnosis of Lyme neuroborreliosis requires laboratory investigation of the spinal fluid including intrathecal antibody production, and the remaining disease manifestations require testing for serum antibodies to B. burgdorferi. Testing individuals with non-specific subjective symptoms is not recommended, because of a low positive predictive value.

Comparison of findings for patients with Borrelia garinii and Borrelia afzelii isolated from cerebrospinal fluid.

BACKGROUND: The most common cause of Lyme neuroborreliosis in Europe is Borrelia garinii, followed by Borrelia afzelii. However, no series describing patients with culture-confirmed cases of Lyme neuroborreliosis have been published, and no comparison of findings for patients with B. garinii and B. afzelii isolated from cerebrospinal fluid (CSF) has been reported. METHODS: All adult patients identified at a single medical center during a 10-year period who had borreliae isolated from CSF and typed as B. garinii or B. afzelii (using large DNA fragment patterns obtained with the MluI restriction endonuclease and separated with pulsed-field gel electrophoresis) were included. RESULTS: A comparison of 23 patients who had B. garinii isolated from CSF with 10 patients who had B. afzelii isolated from CSF revealed that a reliable clinical diagnosis of Lyme neuroborreliosis (before obtaining a CSF culture and intrathecal borrelial antibody production result) was established more frequently in the B. garinii group than in the B. afzelii group (19 of 23 patients vs. 1 of 10 patients). Patients in the B. garinii group reported radicular pains and expressed meningeal signs more often, but reported dizziness less often (occurrences of several other symptoms and/or signs were comparable). Lymphocytic pleocytosis, as well as several other CSF abnormalities, were frequent among patients with B. garinii isolated from CSF but were rare among patients in the B. afzelii group. CONCLUSIONS: Patients with B. garinii isolated from their CSF have a distinct clinical presentation, compared with patients with B. afzelii. B. garinii causes what, in Europe, is appreciated as typical early Lyme neuroborreliosis (Bannwarth syndrome), whereas the clinical features associated with B. afzelii are much less specific and more difficult to diagnose.

Molecular subtyping of Borrelia burgdorferi sensu lato isolates from five patients with solitary lymphocytoma.

Solitary lymphocytoma is a rare cutaneous manifestation of Lyme borreliosis that has been reported almost exclusively from Europe. This suggests that its etiologic agent may be absent or extremely rare on the North American continent. All three species of B. burgdorferi sensu lato known to be associated with human Lyme borreliosis (B. burgdorferi sensu stricto, B. garinii, and B. afzelii have been isolated in Europe, whereas only B. burgdorferi sensu stricto has been found in North America. This suggests that either B. garinii or B. afzelii might be the etiologic agent of borrelial lymphocytoma. To investigate this hypothesis we characterized five strains of B. burgdorferi sensu lato isolated from lymphocytoma lesions of patients residing in Slovenia. The methods used included: large restriction fragment pattern analysis of restriction enzyme MluI-digested genomic DNA, plasmid profiling, protein profiling, ribotyping using 5S, 16S, and 23S rDNA probes, and polymerase chain reaction amplification of the rrf (5S)-rrl (23S) intergenic spacer region. Molecular subtyping showed that four of the five isolates belonged to the species B. afzelii; however, this species is the predominant patient isolate in Slovenia and, therefore, may not represent a preferential association with lymphocytoma. The fifth isolate appeared to be most closely related to the DN127 genomic group of organisms. Further characterization of the isolate revealed that it possessed a unique molecular "fingerprint." The results not only show that borrelial lymphocytoma can be caused by B. afzelii but also demonstrate an association with another genomic group of B. burgdorferi sensu lato that is present in North America as well.

Identification of three species of Borrelia burgdorferi sensu lato (B. burgdorferi sensu stricto, B. garinii, and B. afzelii) among isolates from acrodermatitis chronica atrophicans lesions.

In Europe, at least three species of Borrelia are known to be causative agents of Lyme borreliosis: B. burgdorferi sensu stricto, B. garinii, and B. afzelii. Observable differences in the molecular characteristics of the three species have led to speculation that they may also differ in their pathogenic potential and/or tissue tropisms. Several studies have found an association between the chronic skin manifestation of Lyme borreliosis, acrodermatitis chronica atrophicans, and infection by B. afzelii. We sought to find further evidence for such a correlation by studying the genetic profiles of 22 strains of B. burgdorferi sensu lato derived from 21 patients who presented to the University Medical Center, Ljubljana, Slovenia between 1992 and 1995. Strains were isolated in culture from skin biopsies of acrodermatitis chronica atrophicans lesions; in the case of one patient two separate acrodermatitis chronica atrophicans lesions were cultured. All 21 patients had clinically typical lesions with "classic" histopathology and high IgG antibody titers to B. burgdorferi sensu lato. Strains were characterized and typed by 16S ribosomal RNA-specific polymerase chain reaction and determination of their large restriction fragment patterns using pulsed-field gel electrophoresis of MluI-digested genomic DNA. Of the 22 isolates studied, 17 possessed the highly conserved MLa1 pattern characteristic of B. afzelii. The remaining five isolates possessed large restriction fragment patterns that were typical of B. garinii (MLg2, four isolates from three patients) and B. burgdorferi sensu stricto (MLb2, one isolate). The results of 16S ribosomal RNA-specific polymerase chain reaction were concordant with these species designations. These data show that B. afzelii is the predominant, but not the exclusive, etiologic agent of acrodermatitis chronica atrophicans.

Analysis of Borrelia burgdorferi sensu lato isolated from cerebrospinal fluid.

Involvement of the nervous system in Lyme borreliosis may occur with or without erythema migrans and it may present with a variety of neurological symptoms. In this study we analysed phenotypic and genotypic characteristics of 40 Borrelia strains isolated from cerebrospinal fluid (CSF) of 38 Slovenian patients with different clinical manifestations of Lyme borreliosis. In seven of the patients, Borreliae were also isolated from skin lesions. Species identification and plasmid profiles were determined by pulsed-field gel electrophoresis and protein profiles by SDS-PAGE. MluI digestion profiles of Borrelia burgdorferi sensu lato DNA showed that 25 (62.5%) isolates were B. garinii, 14 (35%) B. afzelii, and one (2.5%) B. burgdorferi sensu stricto. All strains, except one, possessed a large plasmid and a varying number of smaller plasmids. Three (7.5%) isolates exhibited an unusual plasmid profile, with a large plasmid dimer or three copies of the large plasmid. In protein analyses, all strains expressed OspA protein. OspB was present significantly more often in B. afzelii than B. garinii strains (p=0.0000), while OspC was more often present in B. garinii than B. afzelii strains (p=0.0052). In the seven patients with Borreliae isolated also from the skin, the CSF and skin isolates were identical, either B. garinii (six patients) or B. afzelii (one patient). Species and plasmid heterogeneity as well as antigen diversity could play a role in the pathogenesis of the infection. When combined with our own earlier data, the results suggest species-related organotropism.

Isolation of Borrelia burgdorferi sensu lato from blood of children with solitary erythema migrans.

OBJECTIVES: To establish the frequency of isolation of Borrelia burgdorferi sensu lato from blood of children with solitary erythema migrans (EM) in Europe, to determine the strains of the isolated borreliae and to compare the clinical course and the outcome of the disease according to positive and negative blood culture result. METHODS: In the prospective study we included 134 consecutive patients younger than 15 years with solitary EM, referred to our institution in 1996 and 1997. One milliliter of blood was withdrawn before treatment and cultured in modified Kelly-Pettenkofer medium. Isolated borreliae were typed according to LRFP analysis. Patients were treated with either penicillin V or cefuroxime axetil for 14 days. The posttreatment course was surveyed by follow-up visits during 1 year. RESULTS: B. burgdorferi sensu lato was isolated in 12 of 134 (9%) patients. Eleven blood isolates were typed: 10 were found to be B. afzelii and 1 was Borrelia garinii. Comparison of blood culture-positive and -negative patients revealed no differences in pretreatment characteristics or in posttreatment clinical course. However, worsening of local and/or systemic signs and symptoms at the beginning of antibiotic therapy (Jarish-Herxheimer's reaction) was identified more often in the blood culture-positive than in the blood culture-negative group (5 of 12 vs. 17 of 122, respectively; P = 0.0274). CONCLUSIONS: The isolation rate of B. burgdorferi sensu lato from the blood of children with solitary EM was 9%. The majority of the isolates were B. afzelii. Blood culture-positive patients treated with oral antibiotics were not at greater risk for unfavorable course of the disease than patients with negative blood culture result.

Characterisation of Borrelia burgdorferi sensu lato strains isolated from patients with skin manifestations of Lyme borreliosis residing in Slovenia.

Lyme borreliosis is the most prevalent tick-borne infection in Slovenia. Skin disorders are the most frequent clinical manifestations. The aim of the present study was to assess the phenotypic and genotypic diversity of a large number of human Borrelia burgdorferi sensu lato isolates and to evaluate any association between the isolates and different clinical manifestations. All 103 strains tested were from patients suffering from the skin disorders of Lyme borreliosis. Skin biopsies, cerebrospinal fluid and blood samples from patients were inoculated into modified Kelly Pettenkofer medium. Protein profiles were determined by SDS-PAGE and species identification and plasmid profiles by pulsed-field gel electrophoresis. MluI digestion profiles showed that 87 (84.5%) isolates belonged to B. afzelii, 15 (14.5%) to B. garinii and 1 (1%) to B. burgdorferi sensu stricto. The number of plasmids in each strain varied from three to seven, and the plasmid size ranged from 15 to 65 kb. Four isolates of B. garinii possessed multiple large plasmids and four isolates had a large plasmid dimer (three B. afzelii and one B. garinii). Isolates showed qualitative and quantitative differences in protein expression. The study found differences in the expression of OspB and OspC proteins between B. afzelii and B. garinii strains. OspB was expressed significantly more often by B. afzelii (78 of 87, 89.6%) than by B. garinii (4 of 15, 26.6%) isolates, while OspC protein was expressed significantly more often by B. garinii (14 of 15, 93.3%) than by B. afzelii (51 of 87, 58.6%) isolates. In Slovenia, B. afzelii causes the majority of skin lesions. The isolates investigated showed plasmid and protein diversity. Heterogeneity of the spirochaetes may be important for virulence, and may have implications for pathogenesis and therapy of the infection. Differences in immunodominant proteins also have an important impact on serological testing and vaccine development.

Genotypic and phenotypic characterisation of Borrelia burgdorferi sensu lato strains isolated from human blood.

Lyme borreliosis often presents initially with erythema migrans. Borreliae may disseminate from the primary skin lesion, and different organs and systems could be affected. Borrelia strains were isolated from blood of 70 patients with Lyme borreliosis, including 10 patients from whom borreliae were also isolated from skin. The aim of the present study was to characterise the isolates with regard to their phenotypic and genotypic characteristics. Borreliae were cultivated in MKP medium. Species identification and plasmid profiles were determined by pulsed-field gel electrophoresis (PFGE) and protein profiles by SDS-PAGE. Digestion of Borrelia burgdorferi sensu lato DNA showed 63 (90%) B. afzelii Mla1 and 7 (10%) B. garinii Mlg2. No B. burgdorferi sensu stricto were isolated. Borreliae were isolated from both skin and blood of 10 patients, nine pairs of isolates were identical: seven B. afzelii and two B. garinii. B. afzelii was isolated from the skin and B. garinii from blood of the tenth patient. All but one isolate possessed at least one large plasmid and varying numbers of smaller plasmids. Eight (11.4%) of 70 isolates possessed an unusual plasmid profile (2 of 63 B. afzelii and 6 of 7 B. garinii). Borreliae differed in their protein profiles. OspA and OspB proteins were expressed by all B. afzelii isolates; 85.7% of B. garinii isolates expressed OspA and 71.4% expressed OspB. OspC was expressed by 65% of B. afzelii isolates and all B. garinii isolates. The ratios of B. afzelii and B. garinii isolated from blood and skin were similar. These results do not support the hypothesis that B. garinii has a higher propensity for haematogenous dissemination than B. afzelii. Antigen diversity as well as species and plasmid heterogeneity could play a role in the pathogenesis of the infection, suggesting distinctive strain organotropism.

Clinical distinction between human granulocytic ehrlichiosis and the initial phase of tick-borne encephalitis.

OBJECTIVES: The aim of our study was to establish clinical and laboratory differences between patients with acute human granulocytic ehrlichiosis (HGE) and patients with the initial phase of tick-borne encephalitis (TBE). METHODS: Clinical features and laboratory results of four patients with acute HGE (established by the presence of the specific DNA sequences of the HGE agent in whole blood by polymerase chain reaction and/or by seroconversion to the HGE agent by indirect immunofluorescence assay) and 12 patients with the initial phase of TBE (demonstrated by the presence of serum IgM antibodies to TBE virus) were compared. All these patients were uncovered at the Department of Infectious Diseases, University Medical Centre, Ljubljana, Slovenia, during 1995-1996, in a prospective study on the aetiology of febrile illnesses occurring within 6 weeks after a tick bite. RESULTS: Findings were similar for the majority of the examined parameters including severity of illness, level and duration of fever, presence of headache, leukopenia, thrombocytopenia, and liver - function test abnormalities. Statistically significant differences were found only for arthralgia (P=0.026) and elevated concentration of C-reactive protein (P=0.003); both variables were found more often in patients with acute HGE. CONCLUSIONS: In a patient residing in the central part of Slovenia, who reports a tick bite followed by a febrile illness with leukopenia and/or thrombocytopenia, the presence of arthralgias and/or an elevated C-reactive protein value directs toward the diagnosis of acute HGE and against the initial phase of TBE.