Legionella pneumophila—epidemiology and characterization of clinical isolates, Slovenia, 2006–2020

Legionella pneumophila is the causative agent of severe Legionnaires’ disease (LD). Although an increasing number of LD cases have been observed, published data from Slovenia are very limited and data on molecular epidemiology are even scarcer. The present retrospective study (2006–2020) reports the results of the microbiological diagnosis of LD, as well as the epidemiology and characterization of the Legionella clinical isolates. We tested urine samples from 15,540 patients with pneumonia symptoms for L. pneumophila infection by urine antigen test, of which 717 (4.6%) tested positive. Isolation of L. pneumophila was successfully performed from 88 clinical specimens, with 82 (93.2%) being identified as L. pneumophila sg 1 and six (6.8%) as L. pneumophila sg 2–14. Sequence-based typing (SBT) identified 33 different sequence types (STs), the most frequent being ST1 and ST23. Sequence type 1 mainly comprised isolates belonging to the Philadelphia subgroup, and ST23 mostly to Allentown/France. The standard SBT scheme, as well as Dresden phenotyping for L. pneumophila, presented a high diversity among isolates

Leptospirosis and characterization of Leptospira isolates from patients in Koprivnica-Križevci County, Croatia from 2000–2004

Introduction. Leptospirosis, caused by spirochetes of the genus Leptospira, is present in the Koprivnica-Križevci County area, Croatia. Clinical manifestation can range from asymptomatic, short-term mild, non-specific febrile disease, to severe forms with high mortality rates. Aim. The aim of the study was to valuate culture in front of microscopic agglutination test (MAT) for diagnosis of infection, and to evaluate clinical and laboratory features of the disease. Moreover, we want to characterize Leptospira strains involved in infection in Koprivnica-Križevci County, Croatia. Methods. We included 68patients with clinical presentation consistent with leptospirosis collected in a 5-year period (2000–2004). Clinical samples (blood, urine and cerebrospinal fluid, CSF) were inoculated in Kolthoff’s mediumspecies of isolated Leptospira strains was determined with Tm of real-time PCR, serogroup/serovar with MAT and NotI-RFLP analysis. Demonstration of specific antibodies in patients’ sera was done using microscopic agglutination test. Results. Leptospira was isolated from the blood of 14/51(27.5%) patients and the most often identified serogroup/serovar was Icterohaemorrhagiae (8/10, 80%) followed by Grippotyphosa (10%). Regarding to species level, 8/10 isolated belonged to L. interrogans sensu stricto and one to L. kirschneri species. MAT was carried out on 51patients with suspected leptospirosis, and was positive in 11/51(21.5%) patients. Most of our patients presented with moderate severe symptoms, were hospitalized from August to October, and were infected mainly during the work or recreation in our county. The frequency of particular clinical features and pathological laboratory findings correlated with the severity of the clinical condition. Conclusions. Leptospirosis can be confirmed microbiologically, culture and MAT contributed almost equally to the diagnosis of infection. Serovar Icterohaemorrhagiae was found as the dominant one, and L. interrogans sensu stricto as dominant species in our county. Epidemiological data shown that leptospirosis occurs seasonally, affects the rural population, and most commonly is presented with moderate severe clinical course

Clinical course, serologic response, and long-term outcome in elderly patients with early Lyme borreliosis

Infected elderly people often present with signs and symptoms that differ from those in younger adults, but data on the association between patient age and presentation of early Lyme borreliosis (LB) are limited. In this study, the association between patient age (18–44 years, young vs. 45–64 years, middle-aged vs. ≥ 65 years, elderly) and disease course, microbiologic characteristics, and the long-term outcome of treatment was investigated prospectively in 1220 adult patients with early LB manifesting as erythema migrans (EM) at a single-center university hospital. Patients were assessed at enrolment and followed-up for 12 months. Age was associated with comorbidities, previous LB, presenting with multiple EM, and seropositivity to borreliae at enrolment. The time to resolution of EM after starting antibiotic treatment was longer in older patients. At 12 months, 59/989 (6.0%) patients showed incomplete response. The odds for incomplete response decreased with time from enrolment (odds ratio (OR) of 0.49, 0.50, and 0.48 for 2-month vs. 14-days, 6-month vs. 2-month, and 12-month vs. 6-month follow-up visits, respectively), but were higher with advancing age (OR 1.57 for middle-aged vs. young, and 1.95 for elderly vs. young), in women (OR 1.41, 95% confidence interval (CI) 1.01–1.96), in patients who reported LB-associated constitutional symptoms at enrolment (OR 7.69, 95% CI 5.39–10.97), and in those who presented with disseminated disease (OR 1.65, 95% CI 1.09–2.51). The long-term outcome of EM was excellent in patients of all age groups. However, older patients had slower resolution of EM and higher odds for an unfavorable outcome of treatment (OR 1.57, 95% CI 1.05–2.34 for middle-aged vs. youngand OR 1.95, 95% CI 1.14–3.32 for elderly vs. young), manifested predominantly as post-LB symptoms. The presence of LB-associated constitutional symptoms at enrolment was the strongest predictor of incomplete response

Antibiotic Use and Long-Term Outcome in Patients with Tick-Borne Encephalitis and Co-Infection with Borrelia Burgdorferi Sensu Lato in Central Europe. A Retrospective Cohort Study

In this retrospective cohort study of patients with tick-borne encephalitis (TBE), the clinical outcome in relation to co-infection with B. burgdorferi sensu lato (s.l.) and, specifically, the effect of antibiotic treatment on clinical outcome in patients with TBE who were seropositive for borreliae but who did not fulfil clinical or microbiologic criteria for proven co-infection, were assessed at a single university medical center in Slovenia, a country where TBE and Lyme borreliosis are endemic with high incidence. Among 684 patients enrolled during a seven-year period from 2007 through 2013, 382 (55.8%) had TBE alone, 62 (9.1%) had proven co-infection with borreliae and 240 (35.1%) had possible co-infection. The severity of acute illness was similar in all the groups. The odds for incomplete recovery decreased during a 12-month follow-up but were higher in women, older patients, and in those with more severe acute illness. Incomplete recovery was not associated with either proven (odds ratio (OR) 1.21, 95% confidence interval (CI) 0.49–2.95; p = 0.670) or possible co-infection (OR 0.95, 95% CI 0.55–1.65; p = 0.853). Among patients with possible co-infection, older patients were more likely to be prescribed antibiotics, but the odds for incomplete recovery were similar in those who received antibiotics and those who did not (OR 0.82, 95% CI 0.36–1.87; p = 0.630), suggesting that routine antibiotic treatment in patients with TBE and possible co-infection may not be warranted

Antibiotic use and long-term outcome in patients with tick-borne encephalitis and co-infection with Borrelia burgdorferi sensu lato in Central Europe

In this retrospective cohort study of patients with tick-borne encephalitis (TBE), the clinical outcome in relation to co-infection with B. burgdorferi sensu lato (s.l.) and, specifically, the effect of antibiotic treatment on clinical outcome in patients with TBE who were seropositive for borreliae but who did not fulfil clinical or microbiologic criteria for proven co-infection, were assessed at a single university medical center in Slovenia, a country where TBE and Lyme borreliosis are endemic with high incidence. Among 684 patients enrolled during a seven-year period from 2007 through 2013, 382 (55.8%) had TBE alone, 62 (9.1%) had proven co-infection with borreliae and 240 (35.1%) had possible co-infection. The severity of acute illness was similar in all the groups. The odds for incomplete recovery decreased during a 12-month follow-up but were higher in women, older patients, and in those with more severe acute illness. Incomplete recovery was not associated with either proven (odds ratio (OR) 1.21, 95% confidence interval (CI) 0.49–2.95p = 0.670) or possible co-infection (OR 0.95, 95% CI 0.55–1.65p = 0.853). Among patients with possible co-infection, older patients were more likely to be prescribed antibiotics, but the odds for incomplete recovery were similar in those who received antibiotics and those who did not (OR 0.82, 95% CI 0.36–1.87p = 0.630), suggesting that routine antibiotic treatment in patients with TBE and possible co-infection may not be warranted

Comparison of Culture-Dependent and Culture-Independent Methods for Routine Identification of Airborne Microorganisms in Speleotherapeutic Caves

The effective identification of bacterial and fungal isolates is essential for microbiological monitoring in environments like speleotherapeutic caves. This study compares MALDI-TOF MS and the OmniLog ID System, two high-throughput culture-based identification methods. MALDI-TOF MS identified 80.0% of bacterial isolates to the species level, while the OmniLog ID System identified 92.9%. However, species-level matches between the methods were only 48.8%, revealing considerable discrepancies. For discrepant results, MALDI-TOF MS matched molecular identification at the genus level in 90.5% of cases, while the OmniLog ID System matched only in 28.6%, demonstrating MALDI-TOF MS’s superiority. The OmniLog ID System had difficulties identifying genera from the order Micrococcales. Fungal identification success with MALDI-TOF MS was 30.6% at the species level, potentially improvable with a customised spectral library, compared to the OmniLog ID System’s 16.7%. Metagenomic approaches detected around 100 times more microbial taxa than culture-based methods, highlighting human-associated microorganisms, especially Staphylococcus spp. In addition to Staphylococcus spp. and Micrococcus spp. as indicators of cave anthropisation, metagenomics revealed another indicator, Cutibacterium acnes. This study advocates a multi-method approach combining MALDI-TOF MS, the OmniLog ID System, culture-based, and metagenomic analyses for comprehensive microbial identification. Metagenomic sampling on nitrocellulose filters provided superior read quality and microbial representation over liquid sampling, making it preferable for cave air sample collection

Effect of Statin Use on the Clinical Manifestations, Laboratory Test Results and Outcome of Lyme Neuroborreliosis

Statins have anti-inflammatory and potentially antimicrobial activity, but whether they have a beneficial effect on the course of infectious diseases is controversial. In this study, we assessed the impact of pre-existing statin use on the course and outcome of Lyme neuroborreliosis manifested as meningoradiculitis (Bannwarth\u27s syndrome). One hundred and twenty three consecutive patients with Bannwarth\u27s syndrome, of whom 18 (14.6%) were being treated with statins, were included in the study. To assess the influence of statin use on the course and outcome of the disease, univariate and multivariable analyses were performed. No statistically significant association was found between statin pre-treatment and the clinical manifestations, laboratory test results, and outcome of Bannwarth\u27s syndrome. In conclusion, pre-existing use of statins did not significantly impact either the clinical presentation or the outcome of Bannwarth\u27s syndrome

Effect of statin use on the clinical manifestations, laboratory test results and outcome of Lyme neuroborreliosis

Statins have anti-inflammatory and potentially antimicrobial activity, but whether they have a beneficial effect on the course of infectious diseases is controversial. In this study, we assessed the impact of pre-existing statin use on the course and outcome of Lyme neuroborreliosis manifested as meningoradiculitis (Bannwarth’s syndrome). One hundred and twenty three consecutive patients with Bannwarth’s syndrome, of whom 18 (14.6%) were being treated with statins, were included in the study. To assess the influence of statin use on the course and outcome of the disease, univariate and multivariable analyses were performed. No statistically significant association was found between statin pre-treatment and the clinical manifestations, laboratory test results, and outcome of Bannwarth’s syndrome. In conclusion, pre-existing use of statins did not significantly impact either the clinical presentation or the outcome of Bannwarth’s syndrome

First Report on <i>Leptospira</i> Species Isolated from Patients in Slovenia

Leptospirosis is an important worldwide zoonosis, and it has also been reported in Slovenia. The cultivation of Leptospira from human material is difficult. Despite that, we successfully isolated 12 human Leptospira strains isolated from patients between 2002 and 2020 and used various methods for the phenotypic and genotypic characterization of the strains, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) using our own MALDI-TOF data library, melting temperature analysis of the amplified lfb1 gene, determination of Leptospira serogroups using rabbit immune sera, NotI-RFLP of the whole Leptospira genome, multilocus sequence typing (MLST) of seven housekeeping genes, and whole-genome sequencing (WGS)-based typing. We confirmed the presence of four pathogenic Leptospira species (L. kirschneri, L. interrogans, L. borgpetersenii, and L. santarosai) and three serogroups: Grippotyphosa, Icterohaemorrhagiae, and Sejroe. MALDI-TOF identified three of seven isolates at the species level and four isolates at the genus level. Serovars of 8 of the 10 strains were determined using NotI-RFLP. MLST showed that the clinical isolates belonged to sequence types ST17, ST110, and ST155. WGS confirmed the analysis of Leptospira strains using conventional methods. In addition, WGS provided better taxonomic resolution for isolate DDA 10944/10