Search

Rejection (nomen rejiciendum) of the name Borreliella and all new combinations therein is being requested on grounds of risk to human health and patient safety (Principle 1, subprinciple 2 and Rule 56a) and violation to aim for stability of names, to avoid useless creation of names (Principle 1, subprinciple 1 and 3) and that names should not be changed without sufficient reason (Principle 9 of the International Code of Nomenclature of Prokaryotes).

Background In September 2018, Burkholderia cepacia complex (BCC) infections in 3 patients associated with exposure to a mouthwash solution (MWS) were reported to the Robert Koch Institute (RKI). As the product was still on the market and the scale of the outbreak was unclear, a nation-wide investigation was initiated. Methods We aimed to investigate BCC infections/colonizations associated with MWS. Hospitals, laboratories, and public health services were informed that BCC isolates should be sent to the RKI. These isolates were typed by pulsed-field gel electrophoresis (PFGE) and whole-genome sequencing (WGS) including development of an ad hoc core genome MLST (cgMLST) scheme. Results In total, 36 patients from 6 hospitals met the case definition, the last patient in November 2018. Twenty-nine isolates from 26 of these patients were available for typing. WGS analysis revealed 2 distinct cgMLST clusters. Cluster 1 (Burkholderia arboris) contained isolates from patients and MWS obtained from 4 hospitals and isolates provided by the manufacturer. Patient and MWS isolates from another hospital were assigned to cluster 2 (B. cepacia). Conclusions The combined clinical, epidemiological, and microbiological investigation, including whole-genome analysis, allowed for uncovering a supraregional BCC outbreak in health care settings. Strains of B. arboris and B. cepacia were identified as contaminating species of MWS bottles and subsequent colonization and putative infection of patients in several hospitals. Despite a recall of the product by the manufacturer in August 2018, the outbreak lasted until December 2018. Reporting of contaminated medical products and recalls should be optimized to protect patients.

In northern Europe, tick-borne diseases such as Lyme borreliosis (LB) and tick-borne encephalitis (TBE) are well known. The actual incidence of Babesia infections, however, has remained elusive. In this study, the prevalence of antibodies against two Babesia spp. was investigated in a cohort of patients that were seropositive for Borrelia (B.) burgdorferi sensu lato (s.l.). Data were compared to a control group of healthy individuals. Sera were collected from 283 individuals residing in the southernmost region of Sweden, Skåne County. Almost one third of the sera were from patients with a confirmed seropositive reaction against B. burgdorferi s.l. All sera samples were assessed for IgG antibodies against Babesia (Ba.) microti and Ba. divergens by indirect fluorescent antibody (IFA) assays. Seropositive IgG titers for at least one of the Babesia spp. was significantly more common (p

Objectives The aim of this study was to evaluate the development and status quo of the quality of high throughput in vitro diagnostic testing for tetanus and diphtheria antitoxin antibody (ATX) concentrations based on external quality assessment (EQA) data. Methods We analyzed manufacturer-specific data of 22 EQA surveys—each for the detection of tetanus and diphtheria ATX—to check the diagnostic strength of the corresponding in vitro diagnostic systems. Results While the results were mostly well aligned, individual surveys showed widely dispersed ATX concentrations. The medians of manufacturer collectives deviated from the overall median by up to 8.9-fold in the case of diphtheria ATX and by up to 3.5-fold in the case of tetanus ATX. Such a distribution in the results is particularly critical in the cut-off range for immunity and may lead to an incorrect assessment of vaccination status. Conclusion These results were surprising as there are International Standards for both ATX; however, the results may be linked to the high ATX concentration of the reference material, which deviates considerably from clinically significant concentrations. To increase the accuracy and diagnostic strength of both assays, we recommend a recalibration of the test systems and verification of their traceability to the International Standards.

Background Colistin is still a widely used antibiotic in veterinary medicine although it is a last-line treatment option for hospitalized patients with infections caused by multidrug-resistant Gram-negative bacteria. Colistin resistance has gained additional importance since the recent emergence of mobile colistin resistance (mcr) genes. In the scope of a study on colistin resistance in clinical Escherichia coli isolates from human patients in Germany we characterized the mcr-1 gene variants. Results Our PCR-based screening for mcr-carrying E. coli from German patients revealed the presence of mcr-1-like genes in 60 isolates. Subsequent whole-genome sequence-based analyses detected one non-synonymous mutation in the mcr-1 gene for two isolates. The mutations were verified by Sanger sequencing and resulted in amino acid changes Met1Thr (isolate 803-18) and Tyr9Cys (isolate 844-18). Genotyping revealed no relationship between the isolates. The two clinical isolates were assigned to sequence types ST155 (isolate 803-18) and ST69 (isolate 844-18). Both mcr-1 variants were found to be located on IncX4 plasmids of 33 kb size; these plasmids were successfully conjugated into sodium azide resistant E. coli J53 Azir in a broth mating experiment. Conclusions Here we present the draft sequences of E. coli isolate 803-18 carrying the novel variant mcr-1.26 and isolate 844-14 carrying the novel variant mcr-1.27. The results highlight the increasing issue of transferable colistin resistance.

Rejection (

Human babesiosis is reported throughout the world and is endemic in the northeastern and northern Midwestern United States and northeastern China. Transmission is primarily through hard bodied ticks. Most cases of severe disease occur in immunocompromised individuals and may result in prolonged relapsing disease or death.We provide a summary of evidence supporting current treatment recommendations for immunocompetent and immunocompromised individuals experiencing babesiosis.Most cases of human babesiosis are successfully treated with atovaquone and azithromycin or clindamycin and quinine. Severe disease may require prolonged treatment.

Infective endocarditis (IE) is a life-threatening disease that is associated with high morbidity and mortality. Its long-term prognosis strongly depends on a timely and optimized antibiotic treatment. Therefore, identification of the causative pathogen is crucial and currently based on blood cultures followed by characterization and susceptibility testing of the isolate. However, antibiotic treatment starting prior to blood sampling or IE caused by fastidious or intracellular microorganisms may cause negative culture results. Here we investigate the additional diagnostic value of broad-range PCR in combination with direct sequencing on resected heart tissue or swabs in patients with tissue or swab culture-negative IE in a routine clinical setting. Sensitivity, specificity, and positive and negative predictive values of broad-range PCR from diagnostic material in our patients were 33.3%, 76.9%, 90.9%, and 14.3%, respectively. We identified a total of 20 patients (21.5%) with tissue or culture-negative IE who profited by the additional application of broad-range PCR. We conclude that broad-range PCR on resected heart tissue or swabs is an important complementary diagnostic approach. It should be seen as an indispensable new tool for both the therapeutic and diagnostic management of culture-negative IE and we thus propose its possible inclusion in Duke’s diagnostic classification scheme.

We describe a 79-year-old Irish man who, because he had hyposplenism and splenic atrophy due to adult celiac disease, became critically ill from a severe Babesia divergens infection. Greater awareness of the possible consequences of splenic dysfunction from adult celiac disease, such as serious pneumococcal infections and babesiosis, is warranted.

This review is directed at physicians and laboratory personnel in private practice and clinics who treat and diagnose Lyme borreliosis (LB) in patients as part of their daily work. A major objective of this paper is to bring together background information on Borrelia (B.) burgdorferi sensu lato (s.l.) and basic clinical knowledge of LB, which is one of the most frequently reported vector-borne diseases in the Northern Hemisphere. The goal is to provide practical guidance for clinicians and for laboratory physicians, and scientists for a better understanding of current achievements and ongoing obstacles in the laboratory diagnosis of LB, an infectious disease that still remains one of the diagnostic chameleons of modern clinical medicine. Moreover, in bringing together current scientific information from guidelines, reviews, and original papers, this review provides recommendations for selecting the appropriate tests in relation to the patient's stage of disease to achieve effective, stage-related application of current direct and indirect laboratory methods for the detection of B. burgdorferi s.l. Additionally, the review aims to discuss the current state of the art concerning the diagnostic potential and limitations of the assays and test methods currently in use to optimize LB patient management and provide insight into the possible future prospects of this rapidly changing area of laboratory medicine.

To date, relatively little is known about the economic and medical impact of Lyme borreliosis (LB) on European health care systems, especially for the inpatient sector. This retrospective analysis is based on data provided for the years 2007-2011 by a German statutory health insurance company (DAK-Gesundheit) covering approximately 6 million insured. Total cost was calculated for a 1-year period both from the third-party payers and from the societal perspective, respectively. In our cohort the incident diagnosis of LB was coded for 2163 inpatient cases during the years 2008-2011. The median inpatient time was 9 days resulting in a median direct medical cost per hospital stay of 3917€ for adolescents and 2843€ for adults. Based on extrapolation of our findings to the German population, we would expect an average hospital admission of 5200 adults and 2300 adolescents (

The ongoing Libyan conflict constantly causes victims among the military and civilian population. Cross-border transfer of patients represents a high risk of introducing multidrug-resistant organisms (MDROs), for example, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci, and carbapenem-resistant gram-negative organisms (CROs), into the country of destination. This study assessed the MDRO status in Libyan war casualties (n = 67) admitted to Northwest Medical Centre in Frankfurt/Main, Germany, from August 2016 till January 2017. Identified multidrug-resistant nonfermenters and Enterobacteriaceae were subjected to molecular detection of β-lactamases and further mechanisms of resistance. All isolates were typed by enzymatic macrorestriction and subsequent pulsed-field gel electrophoresis. MDROs were found in 40 (60%) patients, including 25 (37%) positive for at least one CRO and 11 (16%) patients with MRSA. A total of 37 isolates of Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Enterobacter cloacae, and Serratia marcescens produced carbapenemases: NDM (n = 17), OXA-48 (n = 15), and OXA-23 (n = 9) in addition to other β-lactamases (with bla

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 Lyme borreliosis 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 Lyme borreliosis. 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 Lyme borreliosis 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 for, and the remaining disease manifestations require testing for antibodies to Borrelia burgdorferi . Testing individuals with non-specific subjective symptoms is not recommended, because of a low positive predictive value.

In order to assess the level of occupational exposure to the main pathogens transmitted by the Ixodes ricinus tick, a seroprevalence study was performed on serum samples collected in 2003 from 2975 forestry workers of northeastern France. The global seroprevalence estimated for the seven pathogens studied was 14.1% (419/2975) for Borrelia burgdorferi sl, 5.7% (164/2908) for Francisella tularensis, 2.3% (68/2941) for tick-borne encephalitis virus, 1.7% (50/2908) for Anaplasma phagocytophilum and 1.7% (48/2908) for Bartonella henselae. The seroprevalences of Babesia divergens and Babesia microti studied in a subgroup of participants seropositive for at least one of these latter pathogens were 0.1% (1/810) and 2.5% (20/810), respectively. Borrelia burgdorferi sl seroprevalence was significantly higher in Alsace and Lorraine and F. tularensis seroprevalence was significantly higher in Champagne-Ardenne and Franche-Comte. The results of this survey also suggest low rates of transmission of Bartonella henselae and F. tularensis by ticks and a different west/east distribution of Babesia species in France. The frequency and potential severity of these diseases justify continued promotion of methods of prevention of I. ricinus bites. E. Rigaud, (C) 2016 European Society of Clinical Microbiology and Infectious Diseases.

While flagella-independent motility has long been described in representatives of the genus Acinetobacter, the mechanism of motility remains ambiguous. Acinetobacter baumannii, a nosocomial pathogen appearing increasingly multidrug-resistant, may profit from motility during infection or while persisting in the hospital environment. However, data on the frequency of motility skills among clinical A. baumannii isolates is scarce. We have screened a collection of 83 clinical A. baumannii isolates of different origin and found that, with the exception of one isolate, all were motile on wet surfaces albeit to varying degrees and exhibiting differing morphologies. Screening a collection of transposon mutants of strain ATCC 17978 for motility defects, we identified 2 akinetic mutants carrying transposon insertions in the dat and ddc gene, respectively. These neighbouring genes contribute to synthesis of 1,3-diaminopropane (DAP), a polyamine ubiquitously produced in Acinetobacter. Supplementing semi-solid media with DAP cured the motility defect of both mutants. HPLC analyses confirmed that DAP synthesis was abolished in ddc and dat mutants of different A. baumannii isolates and was re-established after genetic complementation. Both, the dat and ddc mutant of ATCC 17978 were attenuated in the Galleria mellonella caterpillar infection model. Taken together, surface-associated motility is a common trait of clinical A. baumannii isolates that requires DAP and may play a role in its virulence.

Objectives: To investigate the genetic environment of the metallo-b-lactamase gene blaNDM-1 in an Acinetobacter baumannii isolated in 2007 in a German hospital. Methods: Antimicrobial susceptibility testing was performed and resistance genes were characterized by PCR amplification and sequencing. Transferability of b-lactam resistance was tested by broth mating assays and transformation of plasmids. The genetic background of blaNDM-1 was analysed by primer walking. Typing of the A. baumannii strain was performed by repetitive extragenic palindromic sequence-based PCR (rep-PCR) using the DiversiLab system. Results: The multidrug-resistant A. baumannii isolate harboured b-lactamase genes blaNDM-1 and intrinsic blaOXA-64, but without the insertion sequence ISAba1 often located upstream. Transfer of carbapenem resistance by conjugation and transformation failed. Hybridization of isolated plasmid DNA with blaNDM probes was not successful. Shotgun cloning of whole genomic DNA and sequence analyses revealed that blaNDM-1 was located between two insertion elements of ISAba125. Furthermore, this blaNDM-1-containing transposon structure was integrated into a chromosomal gene encoding a putative A. baumannii major facilitator superfamily (MFS) metabolite/H + symporter. Conclusions: The metallo-b-lactamase gene blaNDM-1 in this A. baumannii strain was integrated in the chromosome on a new transposon structure composed of two copies of insertion sequence ISAba125. The variability of the genetic environment of blaNDM-1 likely facilitates the rapid dissemination of this gene within many Gramnegative bacterial species.

Zusammenfassung In Europa zählen Viren zu den häufigsten Verursachern einer Myokarditis. Im Unterschied zur Perikarditis ist die Symptomatik der Myokarditis oft uncharakteristisch und erfordert den Einsatz von Laboruntersuchungen. Die Abklärung der Virusätiologie begnügt sich meist mit dem Nachweis einer zeitgleich ablaufenden Infektionskrankheit (Plausibilitätsdiagnose). Zur direkten Virusdetektion ist die Entnahme einer Herzbiopsie erforderlich. An diesem Material kann das Virus mittels immunhistologischer und molekularbiologischer Methoden unter gleichzeitiger Beurteilung des inflammatorischen Prozesses nachgewiesen werden. Der Einsatz der verschiedenen Untersuchungsmethoden richtet sich nach dem Kosten-Nutzen-Verhältnis.

Zusammenfassung Entzündliche Herzerkrankungen betreffen kombiniert oder isoliert den Herzmuskel und dessen Hülle. Endo-, Myo- und/oder Perikarditiden haben viele verschiedene Ursachen. Sie verlaufen als akute oder chronische Erkrankung. Neben Viren, die gegenwärtig als auslösende Agentien dominieren, sind weiterhin Bakterien, Pilze und Parasiten anzuführen. Autoimmunologische Prozesse sowie bestimmte Therapeutika, z.B. Cocain, gelten als Auslöser nicht infektiöser Myokarditiden. In 25% der Fälle findet sich bei bestehender Myokarditis eine Perikardbeteiligung. Nachfolgend sollen wichtige mikrobiologische Erreger und deren Nachweismöglichkeiten vorgestellt werden, die im Zusammenhang mit einer Myo- und/oder Perikarditis stehen.

Babesiosis is a zoonosis caused by tick-transmitted intraerythrocytic protozoa of the Phylum Apicomplexa. The disease mostly occurs in the USA, but cases have also been reported in several European countries, in Egypt, India, Japan, Korea, Taiwan, and South Africa. The main pathological event is lysis of erythrocytes resulting in haemolytic anaemia, which in severe cases may lead to organ failure and death, particularly in immunocompromised patients. The 2 groups of parasites involved, Babesia microti-like and Babesia sensu stricto (s.s.) species, differ in their life cycle characteristics and susceptibility to antibabesial drugs. Molecular taxonomy is now making a major contribution to the identification of novel pathogens within both groups. Effective treatment of severe cases was initially hampered by the lack of specific antibabesial drugs for human use, but increased use of supportive measures and of the recently developed antimalarial, atovaquone, particularly in combination with azithromycin, has improved the prospects for management of acute disease especially when caused by Babesia s.s. species. Prevention should be based primarily on increasing the awareness of physicians and the public to the risks, but infection from blood transfusions is particularly difficult to prevent. Expanding deer populations, resulting in wider distribution and greater abundance of ticks, heightened medical awareness, and growing numbers of immunocompromised patients are likely to result in a continuing rise of reported cases.

Tigecycline is a new glycylcycline that has recently been revealed to be very effective in vitro against a variety of Gram-negative and Gram-positive bacteria including multi-drug resistant microorganisms. Using a standardized microdilution susceptibility testing method, we determined the minimal inhibitory concentrations (MICs) and the minimal bactericidal concentrations (MBCs) of tigecycline, in parallel with doxycycline, tetracycline, and other antibiotic agents relevant for Lyme borreliosis treatment such as ceftriaxone and cefotaxime. The activity of all agents against 16 different Borrelia isolates belonging to all borrelial genospecies known to be pathogenic for humans was investigated and analyzed under standardized conditions. The overall rank order of MIC 90 s was tigecycline (≤0.016 mg/L) > ceftriaxone (0.03 mg/L) > cefotaxime (≤0.125 mg/L) > doxycycline (0.25 mg/L) > tetracycline (0.25 mg/L). The rank order of MBC 90 s was tigecycline (0.5 mg/L) > ceftriaxone (2 mg/L) > tetracycline (16 mg/L) > doxycycline (16 mg/L) > cefotaxime (>16 mg/L). High in vitro activity of the new glycylcycline against Borrelia was further substantiated by time-kill experiments performed with B. afzelii isolate EB1. Parallel testing of tigecycline and ceftriaxone demonstrated a bacteriostatic effect for 0.016 mg/L of tigecycline and for 0.03 mg/L for ceftriaxone after 72 h of incubation. Moreover, tigecycline was bactericidal at a concentration of 0.25 mg/L showing a >3 log 10 unit reduction of the initial inoculum, whereas for ceftriaxone a concentration of 2 mg/L was needed.

Previous studies using small numbers of serum samples from human patients and experimentally infected animals identified the frequent presence of antibodies recognizing RevA, a borrelial fibronectin-binding outer surface protein. We now demonstrate that most examined Lyme disease spirochetes from North America and Europe contain genes encoding RevA proteins, some with extensive regions of conservation and others with moderate diversity. Line blot analyses using recombinant RevA from two diverse Lyme disease spirochetes of RevA and serum samples from culture-confirmed human Lyme disease patients from the United States ( n = 46, mainly with early Lyme disease) and Germany (>500, with early and late manifestations of Lyme disease) were performed. The results indicated that a sizable proportion of patients produced antibodies that recognized recombinant RevA. Overall, RevA-based serological studies were less sensitive and less specific than other assay types, such as the VlsE-based C6 peptide assay. However, sera from patients in the initial stages of Lyme disease contained antibodies against RevA, demonstrating that this protein is expressed early in human infection. Thus, RevA may be a useful target for preventative or curative therapies.

OBJECTIVES: To evaluate the potential improvement of antimicrobial treatment by utilizing a new multiplex polymerase chain reaction (PCR) assay that identifies sepsis-relevant microorganisms in blood. DESIGN: Prospective, observational international multicentered trial. SETTING: University hospitals in Germany (n = 2), Spain (n = 1), and the United States (n = 1), and one Italian tertiary general hospital. PATIENTS: 436 sepsis patients with 467 episodes of antimicrobial treatment. METHODS: Whole blood for PCR and blood culture (BC) analysis was sampled independently for each episode. The potential impact of reporting microorganisms by PCR on adequacy and timeliness of antimicrobial therapy was analyzed. The number of gainable days on early adequate antimicrobial treatment attributable to PCR findings was assessed. MEASUREMENTS AND MAIN RESULTS: Sepsis criteria, days on antimicrobial therapy, antimicrobial substances administered, and microorganisms identified by PCR and BC susceptibility tests. RESULTS: BC diagnosed 117 clinically relevant microorganisms; PCR identified 154. Ninety-nine episodes were BC positive (BC+); 131 episodes were PCR positive (PCR+). Overall, 127.8 days of clinically inadequate empirical antibiotic treatment in the 99 BC+ episodes were observed. Utilization of PCR-aided diagnostics calculates to a potential reduction of 106.5 clinically inadequate treatment days. The ratio of gainable early adequate treatment days to number of PCR tests done is 22.8 days/100 tests overall (confidence interval 15-31) and 36.4 days/100 tests in the intensive care and surgical ward populations (confidence interval 22-51). CONCLUSIONS: Rapid PCR identification of microorganisms may contribute to a reduction of early inadequate antibiotic treatment in sepsis.

Objective: Evaluation of the technical and diagnostic feasibility of commercial multiplex real-time polymerase chain reaction (PCR) for detection of blood stream infections in a cohort of intensive care unit (ICU) patients with severe sepsis, performed in addition to conventional blood cultures. Design: Dual-center cohort study. Setting: Surgical ICU of two university hospitals. Patients and participants: One hundred eight critically ill patients fulfilling the American College of Chest Physicians/Society of Critical Care Medicine (ACCP/SCCM) severe sepsis criteria were included. Interventions: None. Measurements and results: PCR results obtained in 453 blood samples from 108 patients were compared with corresponding blood culture results. PCR resulted in a twofold higher positivity rate when compared with conventional blood culture (BC) testing (114 versus 58 positive samples). In 40 out of 58 PCR positive assays the results of the corresponding blood cultures were identical to microorganisms detected by PCR. In 18 samples PCR and BC yielded discrepant results. Compared with conventional blood culture the sensitivity and specificity of PCR was 0.69 and 0.81, respectively. Further evaluation of PCR results against a constructed gold standard including conventional microbiological test results from other significant patient specimen (such as bronchio-alveolar lavage fluid, urine, swabs) and additionally generated clinical and laboratory information yielded sensitivity of 0.83 and specificity of 0.93. Conclusions: Our cohort study demonstrates improved pathogen detection using PCR findings in addition to conventional blood culture testing. PCR testing provides increased sensitivity of blood stream infection. Studies addressing utility including therapeutic decision-making, outcome, and cost-benefit following diagnostic application of PCR tests are needed to further assess its value in the clinical setting

Lyme borreliosis is currently the most frequent tick-transmitted zoonosis in the northern hemisphere. Germany and other European countries are regarded as highly endemic areas; therefore the burden of disease and consequently the costs for the health systems are considered to be high. This report summarises the results of an interdisciplinary workshop on Lyme borreliosis which aimed to identify research deficits and to prioritise areas which need to be addressed. Research needs have been recognised for different areas: diagnosis, epidemiology, immunology, clinics, ecology and health services research. Examples of research areas which have priority are the standardisation of diagnostic tests, the development of markers to detect an active infection, the improvement of the epidemiological database and the analysis of the burden of disease.

The numerous genes and proteins encoded in the borrelial genome have been shown to undergo differential expression in response to environmental cues. To gain a better understanding of possible interactions between antimicrobial agents and Borrelia , we investigated here the effects of increasing concentrations of penicillin G and doxycycline on the protein expression of the Borrelia burgdorferi s.s. isolate LW2 after 24 and 48 h of incubation. For 14 protein spots in Borrelia exposed to penicillin G at 0.25 and 0.5 μg/ml and for 5 protein spots in Borrelia exposed to doxycycline at 0.5 and 1 μg/ml, differences in spot intensity were identified by use of high resolution two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). At concentrations of both antimicrobial agents around the median minimal inhibitory concentration (0.5 μg/ml), all but one of the detected spots showed a considerable down-regulation as revealed by a ⩾50% decrease of spot intensity in comparison to untreated controls. Most of the spots identified thus far belong to proteins that are encoded by genes localized on the borrelial chromosome and are known to be involved in the different pathways of bacterial cell metabolism. Interestingly, one spot, identified as triosephosphate isomerase, was clearly up-regulated in the presence of doxycycline. Our data provide for the first time scientific evidence that B. burgdorferi s.l., although it possesses a small genome and extremely limited biosynthetic capabilities, shows a variable but distinct physiological response to exposure with penicillin G and doxycycline.

Tick-borne infections are among the more important vector-borne infections in the northern hemisphere. However, many facts pertaining to the epidemiology and pathogenesis of such diseases in Europe remain unclear. Human babesiosis in particular may have previously been overlooked in many parts of the world due to a lack of medical awareness and microbiological detection methods. Recently, the first two cases of human babesiosis were reported in Germany, occurring in vicinities where the presence of Babesia spp. in enzootic cycles was obvious for decades but where the risk of acquiring Babesia spp. either from ticks or from human blood products was not known before. It is important to note, though, that as with other tick-borne diseases, Babesia infections may arise in geographic areas where they have not been recorded in the past. Better molecular detection methods and strain typing of parasites are necessary to investigate the epidemiological distribution of zoonotic Babesia spp. in Europe and to clarify whether their virulence or transmissibility is strain-dependent. Therefore, further seroepidemiological and molecular epidemiological studies are urgently needed to learn more about the distribution and medical relevance of these pathogens in various parts of Europe in general and in Germany in particular.

Ever since the discovery of parasitic inclusions in erythrocytes of cattle in Romania by Victor Babes at the end of the 19th century, newly recognised babesial pathogens continue to emerge around the world and the substantial public health impact of babesiosis on livestock and man is ongoing. Babesia are transmitted by ixodid ticks and infection of the host causes a host-mediated pathology and erythrocyte lysis, resulting in anemia, hyperbilirubinuria, hemoglobinuria, and possibly organ failure. Recently obtained molecular data, particularly for the 18S rRNA gene, has contributed significantly to a better understanding of the sometimes puzzling phylogenetic situation of the genus Babesia and new information has been added to help determine the taxonomic position of many species. Moreover, it seems that owing to higher medical awareness the number of reported cases in humans is rising steadily. Hitherto unknown zoonotic babesias are now being reported from geographical areas where babesiosis was not known to occur and the growing numbers of immunocompromised individuals suggest that the frequency of cases will continue to rise. This review covers recent insights into human babesiosis with regard to phylogeny, diagnostics and treatment in order to provide new information on well known as well as recently discovered parasites with zoonotic potential.

Das klinisch variable Erscheinungsbild und die fortbestehenden Schwierigkeiten beim zeitnahen sensitiven und spezifischen laboratoriumsmedizinischen Infektionsnachweis machen die Sepsis immer noch zu einer schwierigen und zumeist primar klinischen Diagnose. Zeitgemase Diagnostikkonzepte fur eine direkte Erregerdiagnose aus Vollblut auf der Basis qualitativer und quantitativer molekularbiologischer Detektionsmethoden werden daher vielfach als ein interessanter Ausweg angesehen, um dem Dilemma einer wenig sensitiven und zumeist relativ zeitintensiven mikrobiellen Erregerdiagnostik auf der Basis klassischer Kulturverfahren zu entgehen und der Notwendigkeit Rechnung zu tragen, bei septischen Patienten moglichst fruhzeitig eine erregerorientierte zielgerichtete Antibiotikatherapie zu initiieren. Zudem legen erste klinische Studienergebnisse die mogliche zukunftige Bedeutung sensitiver kulturunabhangiger Verfahren mit kurzen „Turn-around“-Zeiten fur effektivere Therapiemoglichkeiten und ein besseres Outcome von Patienten mit schweren Infektionen wie Sepsis und septischem Schock nahe. Zunachst sind molekularbiologische Nachweise allerdings trotz vieler Vorteile bei Geschwindigkeit und Sensitivitat als komplementare Verfahren zu sehen und werden klassische Testverfahren wie die Blutkultur auch unter finanziellen Gesichtspunkten in vielen Bereichen nicht kurzfristig ersetzen konnen. Es ist aber damit zu rechnen, dass analog zur rasanten Entwicklung in anderen Technologiefeldern, molekularbiologische Verfahren, die heute noch als kompliziert, arbeitsaufwendig und teuer gelten, in Zukunft den Standard der infektiologischen und mikrobiologischen Diagnostik pragen werden.

Die Babesiose ist eine durch Protozoen hervorgerufene Erkrankung bei Tieren, die durch Zecken auch auf Menschen ubertragen werden kann. Erkrankungen beim Menschen wurden weltweit beschrieben und treten vor allem in Nordamerika, vereinzelt aber auch in Europa auf. Das Krankheitsspektrum reicht von grippeartigen Symptomen bis zu einem malariaahnlichen Syndrom. Bei Patienten mit Splenektomie und/oder Immunsuppression sind schwere, auch todliche Verlaufe bekannt. Ein 63-jahriger Patient mit Zustand nach Splenektomie und Rituximabtherapie bei B-Zell-Lymphom wurde wegen Anamie, Thrombozytopenie, Subikterus, dunklen Urins und Infektzeichen aufgenommen. Laborchemisch zeigten sich eine normochrome Anamie mit positiven Hamolyseparametern und eine Hamoglobinurie mit beginnender Niereninsuffizienz. Im Blutausstrich waren plasmodienahnliche intraerythrozytare Erreger nachweisbar. Es ergaben sich keine Hinweise fur eine Malariaexposition. Der Patient war haufig in den Waldern der Hegau-Bodensee-Region unterwegs, einem bekannten Zeckengebiet. Es wurde die Diagnose einer Babesiose gestellt und mit Polymerase-Kettenreaktion bestatigt. Der Erreger wurde als Babesia EU1 klassifiziert. Die Behandlung mit Clindamycin und Chinin fuhrte nach 4 Wochen zur Remission. Kurz danach trat ein Rezidiv auf, und es wurde eine Langzeittherapie mit Atovaquon und Azithromycin begonnen. Nach mehreren Monaten kam es zur Serokonversion, und die Elimination der Erreger erfolgte schlieslich 8 Monate nach Krankheitsbeginn. Auch in Deutschland ist mit dem Auftreten von Babesiose beim Menschen zu rechnen. Bei Patienten mit Infektzeichen und hamolytischer Anamie muss an eine Babesiose gedacht werden, besonders wenn sie splenektomiert und/oder immunsupprimiert sind.

Lyme borreliosis is a spirochetal infection caused by the Borrelia burgdorferi sensu lato complex that can proceed towards an inflammatory joint manifestation known as Lyme arthritis. Production of chemokines orchestrating neutrophil infiltration is supposed to be key to early arthritic pathogenesis. Using PMA-differentiated macrophage-like THP-1 (mTHP-1) cells we identified by antibody array methodology or mRNA analysis IL-8, GRO-alpha, NAP-2, and SDF-1alpha as being among those chemokines that are upregulated by bacterial lysates obtained from B. burgdorferi. Based on these observations, we set out to characterize in detail mechanisms mediating IL-8 release in this cellular model. TLR2 blocking antibodies, analysis of p65 translocation, and electromobility-shift analysis revealed activation of the TLR2/NF-kappaB axis by B. burgdorferi. The functional importance of this pathway was substantiated by suppression of IL-8 after inhibition of IkappaB kinase. Notably, MAP kinases, specifically the MEK1/2-ERK1/2 pathway, were essential for IL-8 secretion. Those data were confirmed by using freshly isolated adherent peripheral blood mononuclear cells. On the contrary, B. burgdorferi-induced IL-8 in mTHP-1 was unlikely related to flagellin, alpha3beta1-integrin signaling, lipopolysaccharide, bacterial DNA, NOD1/NOD2 agonists, or to intermediate production of IL-1beta and TNF-alpha. Induction of IL-8 by B. burgdorferi was not due to amplification of constitutive AP-1 DNA-binding activity detectable in mTHP-1 cells. Data presented herein validate that TLR2, particularly on mTHP-1 cells, holds a central position in mediating IL-8 secretion associated with extracellular B. burgdorferi and beyond that suggest inhibition of IkappaB kinase and MEK1/2 kinases as promising pharmacological strategies aiming at IL-8 in early Lyme arthritis.

Little is known about the in vitro activity of penems and carbapenems against the spirochete Borrelia burgdorferi. Here, faropenem, ertapenem, imipenem and meropenem as well as the third-generation cephalosporin ceftriaxone and tobramycin were tested in vitro against 11 isolates of the B. burgdorferi sensu lato complex. On a microg/mL basis, ertapenem was the most potent carbapenem (minimal inhibitory concentration (MIC) range: 0.015-0.125 microg/mL), with in vitro activity comparable with that of ceftriaxone against Borrelia. These findings are supported by the results of time-kill experiments in a Borrelia afzelii skin isolate, demonstrating a >3 log10 unit (99.9%) reduction of the inoculum after 96 h of exposure to either drug at a concentration of three log2 unit dilutions above the respective MIC.