Qualitative and quantitative changes in the oral bacterial flora occur shortly after implementation of fixed orthodontic appliances.

INTRODUCTION: In this pilot study, we aimed to determine qualitative and quantitative microbiological changes after the implementation of orthodontic appliances. METHODS: A total of 10 healthy patients aged 12-15 years were recruited who needed to undergo orthodontic treatment with buccal fixed appliances. Gingival conditions were assessed by the Gingival Index, Periodontal Screening Index, and Sulcus Bleeding Index. Microbiological samples were collected before and 1 week after the start of therapy at premolars and molars of the right upper quadrant. Bacterial species were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: The total number of bacteria increased. Six bacterial species were identified that are involved in the development of caries and other infectious processes. The bacteria selectively adapted more efficiently to the new oral milieu compared with the general oral microbial background. There was a significant increase in Streptococcus spp at the premolars and molars. In all individuals, symptoms of inflammation and gingivitis were detected as a response to the bacterial challenge. CONCLUSIONS: Orthodontic treatment induces significant changes in the oral microbial flora associated with gingivitis and an enhanced risk for cariogenic reactions within the first days of orthodontic treatment. To prevent or reduce infectious side effects, oral hygiene instructions and control of patients are necessary before and during the beginning of the therapy.

Preoperative serum pattern analysis to predict the outcome of tonsillectomy in adults with chronic tonsillitis.

The method of classification and tree analysis (CART) was used to predict the outcome of tonsillectomy for chronic tonsillitis (CHT) analyzing patterns of serological markers. In a prospective case study of 24 adult patients with CHT in comparison to 24 patients with acute peritonsillar abscess (PTA) blood samples were assessed 1 day before (T-1) and 3 days after tonsillectomy. Outcome 6 months later (T180) was documented using the Glasgow Benefit Inventory (GBI) and the Specific Benefits from Tonsillectomy Inventory (SBTI). In comparison to PTA, patients with CHT were at best classified by C-reactive protein with a cut-off value of <16.735 mg/dl. For CHT, immunoglobulin E ≤ 144.65 kU/l and the combination of monocytes ≤ 0.565 Gpt/l plus leucocytes >5.855 Gpt/l at T-1 were the best classificators for higher SBTI overall score and symptom score symptom score, respectively, at T180. A higher benefit subscore at T180 was associated to γ-globulin >15.85 % plus α2-globulin >8.950% at T-1. The best classificator for better GBI overall score at T180 was an ASL titer >169.0 IU/ml or the combination of an ASL titer ≤ 169.0 IU/ml with lymphocytes ≤ 2.195 Gpt/l. Lymphocytes ≤ 2.195 Gpt/l were associated with higher GBI general subscore. Leukocytes ≤ 6.780 Gpt/l were related to higher GBI social support subscore. The combination of immunoglobulin A >1.360 g/l with procalcitonin level >0.058 ng/ml was the best combination to classify for higher physical health score. Instead of looking on isolated serologic markers, CART of multiple parameters seems to be more effective to predict the outcome of tonsillectomy for CHT.

Culture independent Raman spectroscopic identification of urinary tract infection pathogens: a proof of principle study.

Urinary tract infection (UTI) is a very common infection. Up to every second woman will experience at least one UTI episode during her lifetime. The gold standard for identifying the infectious microorganisms is the urine culture. However, culture methods are time-consuming and need at least 24 h until the results are available. Here, we report about a culture independent identification procedure by using Raman microspectroscopy in combination with innovative chemometrics. We investigated, for the first time directly, urine samples by Raman microspectroscopy on a single-cell level. In a first step, a database of eleven important UTI bacterial species, which were grown in sterile filtered urine, was built up. A support vector machine (SVM) was used to generate a statistical model, which allows a classification of this data set with an accuracy of 92% on a species level. This model was afterward used to identify infected urine samples of ten patients directly without a preceding culture step. Thereby, we were able to determine the predominant bacterial species (seven Escherichia coli and three Enterococcus faecalis ) for all ten patient samples. These results demonstrate that Raman microspectroscopy in combination with support vector machines allow an identification of important UTI bacteria within two hours without the need of a culture step.

Combined dielectrophoresis-Raman setup for the classification of pathogens recovered from the urinary tract.

Rapid and effective methods of pathogen identifications are of major interest in clinical microbiological analysis to administer timely tailored antibiotic therapy. Raman spectroscopy as a label-free, culture-independent optical method is suitable to identify even single bacteria. However, the low bacteria concentration in body fluids makes it difficult to detect their characteristic molecular fingerprint directly in suspension. Therefore, in this study, Raman spectroscopy is combined with dielectrophoresis, which enables the direct translational manipulation of bacteria in suspensions with spatial nonuniform electrical fields so as to perform specific Raman spectroscopic characterization. A quadrupole electrode design is used to capture bacteria directly from fluids in well-defined microsized regions. With live/dead fluorescence viability staining, it is verified, that the bacteria survive this procedure for the relevant range of field strengths. The dielectrophoretic enrichment of bacteria allows for obtaining high quality Raman spectra in dilute suspensions with an integration time of only one second. As proof-of-principle study, the setup was tested with Escherichia coli and Enterococcus faecalis, two bacterial strains that are commonly encountered in urinary tract infections. Furthermore, to verify the potential for dealing with real world samples, pathogens from patients' urine have been analyzed. With the additional help of multivariate statistical analysis, a robust classification model could be built and allowed the classification of those two strains within a few minutes. In contrast, the standard microbiological diagnostics are based on very time-consuming cultivation tests. This setup holds the potential to reduce the crucial parameter diagnosis time by orders of magnitude.

Genetic characterization and barcoding of taxa in the genus Wolffia Horkel ex Schleid. (Lemnaceae) as revealed by two plastidic markers and amplified fragment length polymorphism (AFLP).

The genus Wolffia of the duckweed family (Lemnaceae) contains the smallest flowering plants. Presently, 11 species are recognized and categorized mainly on the basis of morphology. Because of extreme reduction of structure of all species, molecular methods are especially required for barcoding and identification of species and clones of this genus. We applied AFLP combined with Bayesian analysis of population structure to 66 clones covering all 11 species. Nine clusters were identified: (1) W. angusta and W. microscopica (only one clone), (2) W. arrhiza, (3) W. cylindracea (except one clone that might be a transition form), (4) W. australiana, (5) W. globosa, (6) W. globosa, W. neglecta, and W. borealis, (7) W. brasiliensis, and W. columbiana, (8) W. columbiana, (9) W. elongata. Furthermore, we investigated the sequences of plastidic regions rps16 (54 clones) and rpl16 (55 clones), and identified the following species: W. angusta, W. australiana, W. brasiliensis, W. cylindracea, W. elongata, W. microscopica, and W. neglecta. Wolffia globosa has been separated into two groups by both methods. One group which consists only of clones from North America and East Asia was labelled here "typical W. globosa". The other group of W. globosa, termed operationally "W. neglecta", contains also clones of W. neglecta and shows high similarity to W. borealis. None of the methods recognized W. borealis as a distinct species. Although each clone could be characterized individually by AFLP and plastidic sequences, and most species could be bar-coded, the presently available data are not sufficient to identify all taxa of Wolffia.

Distinct different contributions of the alternative and classical complement activation pathway for the innate host response during sepsis.

Complement activation represents a crucial innate defense mechanism to invading microorganisms, but there is an eminent lack of understanding of the separate contribution of the different complement activation pathways to the host response during sepsis. We therefore investigated different innate host immune responses during cecal ligation and puncture (CLP)-induced sepsis in mice lacking either the alternative (fD(-/-)) or classical (C1q(-/-)) complement activation pathway. Both knockout mice strains showed a significantly reduced survival and increased organ dysfunction when compared with control mice. Surprisingly, fD(-/-) mice demonstrated a compensated bacterial clearance capacity as control mice at 6 h post CLP, whereas C1q(-/-) mice were already overwhelmed by bacterial growth at this time point. Interestingly, at 24 h after CLP, fD(-/-) mice failed to clear bacteria in a way comparable to control mice. However, both knockout mice strains showed compromised C3 cleavage during sepsis. Investigating potential causes for this discrepancy, we were able to demonstrate that despite normal bacterial clearance capacity early during the onset of sepsis, fD(-/-) mice displayed increased inflammatory cytokine generation and neutrophil recruitment into lungs and blood when compared with both control- and C1q(-/-) mice, indicating a potential loss of control over these immune responses. Further in vitro experiments revealed a strongly increased Nf-κB activation capacity in isolated neutrophils from fD(-/-) mice, supporting this hypothesis. Our results provide evidence for the new concept that the alternative complement activation pathway exerts a distinctly different contribution to the innate host response during sepsis when compared with the classical pathway.

Clinical significance and diagnostic usefulness of serologic markers for improvement of outcome of tonsillectomy in adults with chronic tonsillitis.

BACKGROUND: The aim of the present study was to explore serological biomarkers which predict the outcome of tonsillectomy for chronic tonsillitis. METHODS: A case study in a University ENT department of 24 adult patients with chronic tonsillitis (CHT) in comparison to 24 patients with acute peritonsillar abscess (PTA) was performed. Blood samples for clinical routine hematological and serological parameters were assessed prior to surgery (T-1) and five days (T5) after tonsillectomy. Outcome 6 months later (T180) was documented using the Glasgow Benefit Inventory (GBI) and the Specific Benefits from Tonsillectomy Inventory (SBTI). Correlation analyses between CHT and PTA group as well as between the different time points within each group concerning the serological parameters and the outcome parameters were performed. RESULTS: At T-1, patients in the CHT group presented with significantly higher lymphocytes counts (relative and absolute), basophils (relative and absolute) and eosinophils but less white-cells, monocytes, neutrophils (absolute and relative), alpha-1, alpha-2, beta globulins, immunoglobulin and lower C-reactive protein and procalcitonin values than patients in the PTA group (all p < 0.05, respectively). Within each group, different significant changes of the serum parameters (often in opposite direction) were observed between T-1 and T5. SBTI scores at T-1 were significantly lower in the CHT group. In contrast, most GBI scores at T180 were significantly higher in the CHT group. Between T-1 and T180 the SBTI scores improved in three quarters of the CHT patients but only in three fifths of the PTA patients. Higher eosinophil counts and immunoglobulin E levels at T-1 predicted higher GBI scores at T180 in the CHT group. CONCLUSIONS: This pilot study showed a specific serological pattern for patients with chronic tonsillitis with a specific pattern of changes after tonsillectomy. But there is no established role for biomarkers currently used in clinical practice to predict the outcome of tonsillectomy for chronic tonsillitis.

Comparable Studies on Nanoscale Antibacterial Polymer Coatings Based on Different Coating Procedures.

The antibacterial activity of different antibiotic and metal-free thin polymer coatings was investigated. The films comprised quaternary ammonium compounds (QAC) based on a vinyl benzyl chloride (VBC) building block. Two monomeric QAC of different alkyl chain lengths were prepared, and then polymerized by two different polymerization processes to apply them onto Ti surfaces. At first, the polymeric layer was generated directly on the surface by atom transfer radical polymerization (ATRP). For comparison purposes, in a classical route a copolymerization of the QAC-containing monomers with a metal adhesion mediating phosphonate (VBPOH) monomers was carried out and the Ti surfaces were coated via drop coating. The different coatings were characterized by X-ray photoelectron spectroscopy (XPS) illustrating a thickness in the nanomolecular range. The cytocompatibility in vitro was confirmed by both live/dead and WST-1 assay. The antimicrobial activity was evaluated by two different assays (CFU and BTG, resp.,), showing for both coating processes similar results to kill bacteria on contact. These antibacterial coatings present a simple method to protect metallic devices against microbial contamination.

Truncated human cytidylate-phosphate-deoxyguanylate-binding protein for improved nucleic acid amplification technique-based detection of bacterial species in human samples.

A trunk of human cytidylate-phosphate-deoxyguanylate-binding protein/CXXC finger protein 1 (CFP1), immobilized onto an aminohexyl-Sepharose column, can be used as a preanalytical tool for the selective enrichment of bacterial DNA from mixed solutions with high amounts of human background DNA for nucleic acid amplification technique-based detection of pathogens. The transcriptional activator protein exhibits a high affinity for nonmethylated CpG dinucleotide motifs, which are differentially distributed in prokaryotic and higher eukaryotic genomes. The feasibility of the affinity chromatography (AC) step was tested with DNA from severely septic patients. AC using 16S rRNA gene primers substantially increased PCR sensitivity. Approximately 90% of eukaryotic DNA was removed, which significantly increased the signal-to-noise ratio. Threshold cycle values revealed that sensitivity was elevated at least 10-fold. The change in the ratio of bacterial DNA to human DNA increased from 26% to 74% the likelihood of culture-independent PCR-based identification of bacterial presence. Compared to the results seen with blood culture (which is the clinical gold standard for systemic infections, exhibiting 28% positives), the combination of AC and PCR achieves a significant increase in sensitivity and contributes to shortening the time to results for the initiation of guided antibiotic therapy.

Identification of bacterial DNA in neutrocytic and non-neutrocytic cirrhotic ascites by means of a multiplex polymerase chain reaction.

BACKGROUND: Even though bacterial cultures of ascitic fluid are negative in up to 65% of the cases of spontaneous bacterial peritonitis (SBP); bacterial DNA (bactDNA) has been frequently detected in episodes of SBP as well as in culture-negative non-neutrocytic ascites. AIMS: To evaluate multiplex polymerase chain reaction (PCR) for pathogen identification in SBP and to determine the prevalence of ascitic bactDNA and its prognostic relevance in hospitalized patients with liver cirrhosis. METHODS: Ascitic fluid from 68 consecutive patients who underwent diagnostic paracentesis was analysed for polymorphonuclear leucocyte (PMN) count, bacterial culture and bactDNA. BactDNA was identified by gel analysis after multiplex PCR of selectively enriched prokaryotic nucleic acids. Correlations of bactDNA status with PMN count, bacterial culture result and 3-month mortality were determined for neutrocytic and for non-neutrocytic ascites. RESULTS: 11/68 patients presented with an elevated ascitic PMN count. BactDNA was detected in 5/5 culture-positive neutrocytic samples, in 1/6 culture-negative neutrocytic samples and in 8/56 culture-negative non-neutrocytic samples. Three-month mortality did not differ with respect to ascitic bactDNA status (7/14 vs. 14/47, P=0.162). 3-month mortality was increased in the presence of ascitic bactDNA for patients older than 65 years (4/5 vs. 4/14, P=0.046) and for patients with a model for end-stage liver disease score >15 (7/10 vs. 9/30, P=0.025). CONCLUSIONS: Identification of ascitic bactDNA is an appropriate alternative to bacterial ascite culture for pathogen identification in patients at risk for SBP. Its prognostic relevance as a proposed marker of bacterial translocation for certain risk groups has to be further evaluated.

First Time Isolation of Mycobacterium hassiacum From a Respiratory Sample.

We describe the first isolation of Mycobacterium hassiacum, a rapid-growing, partial acid-resistant mycobacterium, in a respiratory specimen from a patient with exacerbated chronic obstructive pulmonary disease. To provide therapeutic recommendation for future cases, antibiotic susceptibility testing of 3 clinical isolates was performed by broth microdilution. All strains tested showed susceptibility to clarithromycin, imipenem, ciprofloxacin, and doxycycline. The role of M hassiacum as a respiratory pathogen remains unclear and needs to be evaluated by future reports.

Ventriculitis due to infection with Rhizopus arrhizus.

A 52-year-old heart-lung transplant patient presented to the emergency department with acute onset of neurologic symptoms. MRI showed ballooning of the left ventricle, midline shift and contrast enhancement in the anterior horn of the left ventricle. Ventricle neuroendoscopy revealed whitish, floccose aerial structures within the left ventricle. Brain biopsy cultures grew Rhizopus arrhizus. Therapy with liposomale amphotericin B and posaconazole was performed. Except for hemianopsia and deficits in minute motor activity, the patient completely recovered.

Superantigen-like gene(s) in human pathogenic Streptococcus dysgalactiae, subsp equisimilis: genomic localisation of the gene encoding streptococcal pyrogenic exotoxin G (speG(dys)).

Streptococcus pyogenes (GAS) causes about 90% of streptococcal human infections while group C (GCS) and G (GGS) streptococci can be pathogenic for different mammalians. Especially the human pathogenic GCS and GGS, Streptococcus dysgalactiae, subsp. equisimilis, account for 5-8% of the human streptococcal diseases like wound infections, otitis media, purulent pharyngitis and also streptococcal toxic shock syndrome. A defined superantigen so far was not identified in GCS and GGS strains. In the present investigation we screened DNA of GCS and GGS human isolates for the presence of genes for streptococcal pyrogenic exotoxins (spe) by hybridisation with probes that stand for the GAS genes speA, speC, speZ (smeZ), speH, speG, speI, speJ and ssa. In many GCS and GGS strains we found positive reactions with the probes speG, speJ and ssa, but not with the probes for the remaining genes under investigation. PCR amplification with subsequent sequence analysis of the PCR fragments revealed only the presence of the gene speG in GCS and GGS strains, while no DNA fragments specific for speJ and ssa could be amplified. Additionally, the upstream and downstream regions flanking speG in GGS strain 39072 were sequenced. Remarkable differences were found in the neighbourhood of speG between GAS and GGS sequences. Downstream of speG we identified in strain GGS 39072 two new open reading frames encoding proteins with no similarity to protein sequences accessible in the databases so far. In the compared GAS strains SF370 and MGAS8232, this segment, apart from some small fragments, had been deleted. Our analysis suggests that a gene transfer from GGS to GAS has preceded following deletion of the two genes orf1 and orf2 in GAS.

Comparison of multilocus sequence typing, RAPD, and MALDI-TOF mass spectrometry for typing of ß-lactam-resistant Klebsiella pneumoniae strains.

Extended spectrum of ß-lactam (ESBL) resistance of Klebsiella pneumoniae has become an increasing problem in hospital infections. Typing of isolates is important to establish the intrahospital surveillance of resistant clones. In this study, the discriminatory potential of randomly amplified polymorphic DNA and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analyses were compared with multilocus sequence typing (MLST) by using 17 ß-lactam-resistant K. pneumoniae isolates of different genotypes. MLST alleles were distributed in 8 sequence types (STs). Among ESBL strains of the same ST, the presence of different ß-lactamase genes was common. RAPD band patterns also revealed 8 types that corresponded to MLST-defined genotypes in 15 out of 17 cases. MALDI-TOF analysis could differentiate 5 clusters of strains. The results of this work show that RAPD may be usable as a rapid screening method for the intrahospital surveillance of K. pneumoniae, allowing a discrimination of clonally related strains. MALDI-TOF-based typing was not strongly corresponding to genotyping and warrants further investigation.

Attributable costs of patients with candidemia and potential implications of polymerase chain reaction-based pathogen detection on antifungal therapy in patients with sepsis.

PURPOSE: The purposes of this study were to calculate attributable costs of candidemia in patients with severe sepsis and to obtain preliminary data regarding the potential effects of polymerase chain reaction-based pathogen detection on antifungal therapy for these patients. METHODS: Patients treated between 2004 and 2010 because of severe sepsis were included into this retrospective analysis. The hospital management provided annual fixed costs per patient-day; data for variable intensive care unit costs were taken from the literature. Multiplex polymerase chain reaction (PCR) was used (VYOO, SIRS-Lab, Jena, Germany) for pathogen detection in the blood. RESULTS: Thirty-two patients with candidemia were identified. Of 874 patients with sepsis, propensity score matching found 32 corresponding patients with sepsis but without candida infection but similar risk factors for developing candidemia. Attributable costs of candidemia were 7713.79 Euro (cost increase, 19.4%). Initiation of antifungal therapy was reduced from 67.5 (52.4, 90) hours in the group, where candida infection was determined by blood culture, to 31.0 (28.0, 37.5; P < .01) hours after detection by multiplex PCR. CONCLUSIONS: Candidemia increases costs of care in patients with septic shock. Polymerase chain reaction-based pathogen detection significantly reduces the time to initiation of antifungal therapy. This might impact on the clinical course of the disease but need to be confirmed in further trials.

Evaluation of a polymerase chain reaction assay for pathogen detection in septic patients under routine condition: an observational study.

BACKGROUND: Treatment of septic shock relies on appropriate antimicrobial therapy. Current culture based methods deliver final results after days, which may delay potentially lifesaving adjustments in antimicrobial therapy. This study was undertaken to compare PCR with blood culture results under routine conditions regarding 1. impact on antimicrobial therapy, and 2. time to result, in patients with presumed sepsis. METHODOLOGY/PRINCIPAL FINDINGS: This was an observational study in a 50 beds ICU of a university hospital. In 245 patients with suspected sepsis, 311 concomitant blood cultures and blood for multiplex PCR (VYOO(®)) were obtained. 45 of 311 blood cultures (14.5%) and 94 of 311 PCRs (30.1%) were positive. However, blood culture or microbiological sampling from the presumed site of infection rarely confirmed PCR results and vice versa. Median time to positivity and interquartile range were 24.2 (18.0, 27.5) hours for the PCR and 68 (52.2, 88.5) hours for BC (p<0.01). PCR median time to result was dependent on technician availability (53.5 hours on Saturdays, 7.2 hours under optimal logistic conditions). PCR results showed good correlation with procalcitonin (p<0.001). In 34% of patients with positive PCRs antimicrobial therapy was considered inadequate according to assessment of clinical arbitrators including 5 patients with vancomycin-resistant enterococci (VRE), 3 cases with multiresistant staphylococci, and 4 patients with fungi. CONCLUSIONS: The results of this observational study support the hypothesis that PCR results are available faster, are more frequently positive, and may result in earlier adjustment of antimicrobial therapy. However, shorter time to result can only be fully exploited when the laboratory is adequately staffed for a 24 hour/7 day service, or when point of care/automated assay systems become available.

Co-circulation of emerging tick-borne pathogens in Middle Germany.

From May until October 2007, a total of 658 Ixodes ricinus ticks were collected off birds (189), rodents (273), and vegetation (196) in a certain area of Middle Germany and investigated for infection with Babesia spp., Anaplasma phagocytophilum, and Rickettsia spp. Overall, 13.1% (86/658) of the ticks were infected with at least one pathogen; co-infections occurred in 0.6% (4/658). Babesia spp. specific DNA was detected in 9.7% (64/658) of the ticks, 1.4% (9/658) were infected with A. phagocytophilum, and 2.6% (17/658) harbored rickettsiae. At least two different Rickettsia species were identified: Rickettsia monacensis and Rickettsia helvetica. Our study provides first interesting insights into the circulation and co-circulation of several emerging pathogens not only in ticks parasitizing birds and small mammals as potential reservoirs but also in questing ticks in a single natural habitat.

Detection of Rickettsia spp. and Anaplasma phagocytophilum in Ixodes ricinus ticks in a region of Middle Germany (Thuringia).

Rickettsia spp. and Anaplasma spp. are regarded as potentially emerging tick-borne pathogens, but so far data on prevalence rates in questing ticks and reports on human diseases in several parts of Europe are rarely available. In this study, 430 nymphs and 570 adult Ixodes (I.) ricinus ticks were collected from a frequently visited forest region of Thuringia (Zeitzgrund, near Stadtroda) in 2006 (n=506) and 2007 (n=494). Individual ticks were investigated for a part of the citrate synthase gene (gltA) of Rickettsia spp. and the 16S rRNA gene of Anaplasma phagocytophilum. Positive amplicons were identified with restriction fragment length polymorphism (RFLP) and/or sequencing. Overall, 14.7% (147/1000) of investigated ticks were infected with Rickettsia spp. After sequencing of 64/147 positive amplicons R. helvetica (29/64) was detected predominantly. Prevalence varied in different developmental stages between 9.3% (40/430) in nymphs and 18.8% (107/570) in adults. A. phagocytophilum-specific DNA was detected in 5.4% (54/1000) of ticks with an infection rate of 4.7% (20/430) in nymphs and 6.0% (34/570) in adults. In 1% (10/1000) of ticks coinfections with Rickettsia spp. and A. phagocytophilum were found. Our study provides interesting insights into the circulation and cocirculation of different rickettsial species and A. phagocytophilum in the same biotope.

Coexistence of Borrelia spp. and Babesia spp. in Ixodes ricinus ticks in Middle Germany.

Altogether, 430 nymphs and 570 adult Ixodes ricinus ticks were collected in 2006 (n = 506) and 2007 (n = 494) from a forest area in Middle Germany (Thuringia). Single ticks were investigated by polymerase chain reaction and restriction fragment length polymorphism or sequencing for Borrelia spp. (ospA gene) and Babesia spp. (18S rRNA gene). Overall, 27.0% (270/1000) were infected with Borrelia species. Out of these, Borrelia garinii was detected most frequently (133/270)-especially, OspA serotype 6 (51/270), followed by Bo. burgdorferi (70/270), Bo. afzelii (42/270), Bo. valaisiana subgroup I (28/270), not typable Borrelia spp. (5/270), Bo. spielmanii (3/270), Bo. valaisiana subgroup II (2/270), and Bo. lusitaniae (1/270). In 1.4% of investigated ticks mixed infections with several Borrelia spp. occurred. Babesia spp.-specific DNA was detected in 5.0% of ticks. Babesia microti was slightly more prevalent (28/50) than Babesia divergens (20/50). Moreover, 5.9% (16/270) of Borrelia spp.-infected ticks were coinfected with Babesia spp. Knowledge on the degree of heterogeneity of Borrelia species and OspA types is prerequisite not only for local risk assessment, but also for diagnostic test and vaccine development.