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Nick Spindler,1 Annette Moter,2,3 Alexandra Wiessner,2,3 Tanja Gradistanac,4 Michael Borger,5 Arne C Rodloff,6 Stefan Langer,1 Judith Kikhney2,3 1Department of Orthopedic Surgery, Traumatology and Plastic Surgery, University Hospital Leipzig, Leipzig, Germany; 2Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health Institute of Microbiology, Infectious Diseases and Immunology, Biofilmcenter, Berlin, Germany; 3MoKi Analytics GmbH, Berlin, Germany; 4Department of Pathology, University Clinic Leipzig, Leipzig, Germany; 5Department of Cardiac Surgery, Leipzig Heart Center, University of Leipzig, Leipzig, Germany; 6Institute of Microbiology and Epidemiology of Infectious Diseases, University Hospital Leipzig, Leipzig, GermanyCorrespondence: Nick SpindlerDepartment of Orthopedic Surgery, Traumatology and Plastic Surgery, University Hospital Leipzig, Leipzig, GermanyTel +49-341 - 9717140Fax +49-341 - 9717139Email nick.spindler@medizin.uni-leipzig.dePurpose: Postoperative mediastinitis after cardiac surgery is still a devastating complication. Insufficient microbiological specimens obtained by superficial swabbing may only detect bacteria on the surface, but pathogens that are localized in the deep tissue may be missed. The aim of this study was to analyze deep sternal wound infection (DSWI) samples by conventional microbiological procedures and fluorescence in situ hybridization (FISH) in order to discuss a diagnostic benefit of the culture-independent methods and to map spatial organization of pathogens and microbial biofilms in the wounds.Methods: Samples from 12 patients were collected and analyzed using classic microbiological culture and FISH in combination with molecular nucleic acid amplification techniques (FISHseq). Frequency of and the time to occurrence of a DSWI was recorded, previous operative interventions, complications, as well as individual risk factors and the microbiologic results were documented.Results: Tissue samples were taken from 12 patients suffering from DSWI. Classical microbiological culture resulted in the growth of microorganisms in the specimens of five patients (42%), including bacteria and in one case Candida. FISHseq gave additional diagnostic information in five cases (41%) and confirmed culture results in seven cases (59%).Conclusion: Microbial biofilms are not always present in DSWI wounds, but microorganisms are distributed in a “patchy” pattern in the tissue. Therefore, a deep excision of the wound has to be performed to control the infection. We recommend to analyze at least two wound samples from different locations by culture and in difficult to interpret cases, additional molecular biological analysis by FISHseq.Keywords: biofilm, FISH, mediastinitis, microorganisms, DSWI

Abstract The epidemiology of Candida infections has changed over the last two decades: The number of patients suffering from such infections has increased dramatically and the Candida species involved have become more numerous as Candida albicans is replaced as an infecting agent by various non-C. albicans species (NAC). At the same time, additional antifungal agents have become available. The different Candida species may vary in their susceptibility for these various antifungals. This draws more attention to in vitro susceptibility testing. Unfortunately, several different test methods exist that may deliver different results. Moreover, clinical breakpoints (CBP) that classify test results into susceptible, intermediate and resistant are controver- sial between CLSI and EUCAST. Therefore, clinicians should be aware that interpretations may vary with the test system being followed by the microbiological laboratory. Thus, knowledge of actual MIC values and pharmacokinetic properties of individual antifungal agents is important in delivering appropriate therapy to patients

Vellore, a region in southern India, has a high incidence of severe human infections with Beta-hemolytic group C and G streptococci (GCGS). To determine the causative species in these infections, we conducted 16S rRNA gene sequencing: Streptococcus dysgalactiae subsp. equisimilis (81%) and S. anginosus (19%) were the causative organisms in the 2-year study period (2006-2007). We used PCR to detect the virulence-related emm gene; results showed that it was restricted to S. dysgalactieae subsp. equisimilis isolates of 99.2% tested positive. Due to a novel marker, S. anginosus and S. constellatus can be quickly and accurately distinguished from other members of the genus. The notable contribution of the anginosus group to human infections suggests that this group of obligate pathogens deserves more attention in healthcare and research. [ABSTRACT FROM AUTHOR]

Background: Recent data show an emergence of resistance in the Bacteroides fragilis group against several antimicrobial agents and inducible resistance against metronidazole in nim-positive strains. The aim of the present study was to investigate inducible metronidazole resistance in nim-positive as well as in nim-negative B. fragilis group strains.Materials and Methods: Of 18 B. fragilis strains (including four nim-positive reference strains and one ATCC strain), two Bacteroides ovatus strains, and one Bacteroides thetaiotaomicron DSM strain minimum inhibitory concentration (MIC) values for metronidazole were determined by Etest and analyzed for nim genes (nimA to-G) by PCR. For this purpose bacterial suspensions were incubated on supplemented Columbia agar plates containing metronidazole at twice the MIC value of the specific strain and incubated under anaerobic conditions for 48 hours. After incubation, growing bacteria were harvested and thereafter incubated at four times the original MIC. This procedure was repeated with increasing antibiotic concentrations. The resulting MIC values were confirmed by Etest.Results: The MIC values for metronidazole of the four nim-positive reference strains ranged from 3 to 8 mg/l. The B. fragilis ATCC 25285 strain and the B. thetaiotaomicron DSM 2255 strain were nim negative with MIC values of 0.19 mg/l and 0.75 mg/l, respectively. Three clinical isolates of B. fragilis strains showed MIC values of > 256 mg/l. In all three strains, nim genes were detected by PCR. The other clinical isolates were nim negative. In these strains, MIC values ranged from 0.19 to 0.75 mg/l. After several passages on metronidazole containing agar, all B. fragilis group strains exhibited MIC values of > 256 mg/l determined by Etest[R].Conclusion: Metronidazole resistance can be selected not only in nim-positive strains but also in nim-negative strains, suggesting that mechanisms other than nim genes are involved. These findings and the emerging resistance of the B. fragilis group against several antimicrobial agents underscore the importance of susceptibility testing of anaerobes even in routine laboratories. [ABSTRACT FROM AUTHOR]

We report on an 8-year-old Haemophilus influenzae type b-vaccinated girl with meningitis due to H. influenzae type f. The girl had an oro-facio-digital syndrome and a hypogammaglobulinemia, obviously predisposing the patient to invasive infections. Treatment with cefotaxime was successful.Incidence, predisposing factors and the influence of H. influenzae type b immunization on invasive infections with non-type b H. influenzae strains in children are discussed. [ABSTRACT FROM AUTHOR]

BACKGROUND: We investigated an outbreak of Serratia marcescens in the neonatal intensive care unit (NICU) and the pediatric intensive care unit (ICU) of the University Children's Hospital Leipzig, Germany. PATIENTS AND METHODS: From September to November 1998 15 patients were infected or colonized by S. marcescens. During the outbreak swabs from eye, blood, throat and nose were taken from every patient hospitalized in the ICUs. RESULTS: In 15 cases (14 from the NICU and one from the pediatric ICU) the cultures yielded S. marcescens. All strains were investigated by pulsed field gel electrophoresis (PFGE) as well as by polymerase chain reaction (PCR) fingerprinting. Both molecular typing methods revealed corresponding fingerprint patterns in all of the 15 isolates. Typing results of the outbreak-related isolates demonstrated that two epidemic strains of distinct genotypes were associated with cross-infections of a group of five and a group of ten patients, respectively. The three invasive and seven of the colonizing isolates were related genotypically. CONCLUSION: This survey shows that PCR and PFGE are comparable in respect to the discrimination and reproducibility for epidemiological studies of S. marcescens strains in nosocomial outbreaks. Genotypic fingerprinting of bacterial isolates is useful and important to limit nosocomial infections. Fingerprinting sources of nosocomial infections can be traced both by PFGE and PCR. All patients infected recovered completely and the nosocomial outbreak could be stopped rapidly. [ABSTRACT FROM AUTHOR]

BACKGROUND: Stenotrophomonas maltophilia is an opportunistic microorganism, often highly resistant to routinely tested antibiotics. This microorganism is isolated in specimens from patients with nosocomial infections with increasing frequency. PATIENTS AND METHODS: During a 1-year period (1998/1999) S. maltophilia was isolated from 137 specimens (0.26% of all investigated specimens) from 80 patients who were treated in a 1,500 bed major tertiary care teaching hospital in Leipzig. The data of 76 patients (133 specimens) could be collected and analyzed completely. RESULTS: The pathogen was most frequently detected in specimens from the respiratory tract (54%). In five patients (six cases) S. maltophilia was isolated from blood cultures (0.3% of all positive blood cultures; 1.4% of all gram-negative isolates from blood cultures). 70 of the infected patients were inpatients and 32 (42%) of them were treated on the internal medicine wards. Of these 32 patients only six (19%) were pretreated with imipenem. The Length of stay at the hospital resulted in an independent increased risk of infection with S. maltophilia. In addition, this organism was detected in six infected outpatients. CONCLUSION: S. maltophilia is not only a nosocomial pathogen. Pretreatment with a carbapenem is no longer an unequivocal risk factor for an infection with S. maltophilia. [ABSTRACT FROM AUTHOR]

A case of Legionella pericarditis caused by a Legionella pneumophila isolate other than serogroup 1 is reported in a 59-year-old man after allogeneic peripheral blood stem cell transplantation. On admission a 5 mm pericardial effusion was detected on echocardiography. Antibodies were detected against L. pneumophila serogroups 7 to 14 using the antigen pool and against serogroup 12 alone. Antibodies were not detected against the serogroup 1 to 6 antigen pool. The patient's clinical condition improved dramatically after treatment with clarithromycin and an echocardiography revealed the total disappearance of the pericardial effusion. [ABSTRACT FROM AUTHOR]

Periprosthetic joint infections (PJI) are serious complications after arthroplasty, associated with high morbidity, mortality, and complex treatment processes. The outcomes of different PJI entities are largely unknown. The aim of this study was to access the early outcomes of different PJI entities. A retrospective, single-center study was conducted. The characteristics and outcomes of patients with PJI treated between 2018 and 2019 were evaluated 12 months after the completion of treatment. Primary endpoints were mortality, relapse free survival (RFS) and postoperative complications (kidney failure, sepsis, admission to ICU). A total of 115 cases were included [19.1% early (EI), 33.0% acute late (ALI), and 47.8% chronic infections (CI)]. Patients with ALI were older ( p = 0.023), had higher ASA scores ( p = 0.031), preoperative CRP concentrations ( p = 0.011), incidence of kidney failure ( p = 0.002) and sepsis ( p = 0.026). They also tended towards higher in-house mortality (ALI 21.1%, 13.6% EI, 5.5% CI) and admission to ICU (ALI 50.0%, 22.7% EI, 30.9% CI). At 12 months, 15.4% of patients with EI had a relapse, compared to 38.1% in ALI and 36.4% in CI. There are differences in patient characteristics and early outcomes between PJI entities. Patients with EI have better early clinical outcomes. Patients with ALI require special attention during follow-up because they have higher occurrences of relapses and postoperative complications than patients with EI and CI.

With extended-spectrum β-lactamases (ESBLs) and CTX-M enzymes being on the rise, antimicrobial treatment of enterobacterial infections is becoming more and more challenging. Our study aimed at a molecular characterization of phenotypically ESBL-positive E. coli strains obtained from blood cultures of patients of the University Hospital of Leipzig (UKL), Germany. The presence of CMY-2, CTX-M-14 and CTX-M-15 was investigated using Streck ARM-D Kit (Streck, USA). Real-time amplifications were performed by QIAGEN Rotor-Gene Q MDx Thermocycler (QIAGEN, Thermo Fisher Scientific, USA). Antibiograms as well as epidemiological data were evaluated. Among 117 cases, 74.4% of the isolates showed a resistance to ciprofloxacin, piperacillin and ceftazidime or cefotaxime while being susceptible to imipenem/meropenem. The proportion of ciprofloxacin resistance was significantly higher than the proportion of ciprofloxacin susceptibility. At least one of the investigated genes was detected in 93.1% of the blood culture E. coli isolates: CTX-M-15 (66.7%), CTX-M-14 (25.6%) or the plasmid-mediated ampC gene CMY-2 (3.4%). 2.6% were tested positive for two resistance genes. 94 of the corresponding stool specimens tested positive for ESBL producing E. coli (94/112, 83.9%). 79 (79/94, 84%) E. coli strains found in the stool samples matched with the respective patient's blood culture isolate phenotypically (MALDI-TOF, antibiogram). The distribution of resistance genes was in accordance with recent studies in Germany as well as worldwide. This study provides indications of an endogenous focus of infection and emphasize the importance of screening programs for high-risk patients., (© 2023. The Author(s).)

Strain 49125 T was isolated from an infant with pneumonia and septicaemia at the Leipzig University Hospital. Phenotypic and genomic traits were investigated. The strain's biochemical profile and its MALDI-TOF spectrogram did not differ from comparative samples of Leclercia adecarboxylata , thus far the sole member of the Leclercia species. A circular genome with a size of 4.4 Mbp and a G+C content of 55.0 mol% was reconstructed using hybrid Illumina and Nanopore sequencing. Phylogenetic analysis was based on 172 marker genes and validated using a k-mer-based search against a large genome collection including subsequent in silico DNA-DNA hybridization. Whole genome average nucleotide identity to any described species was below 95%, suggesting that strain 49125 T represents a new species, for which we propose the name Leclercia pneumoniae sp. nov. with the type strain 49125 T (=LMG 32245 T =DSM 112336 T ).

Actinomyces species play an important role in the pathogenesis of oral diseases and infections. Susceptibility testing is not always routinely performed, and one may oversee a shift in resistance patterns. The aim of the study was to analyze the antimicrobial susceptibility of 100 well-identified clinical oral isolates of Actinomyces spp. against eight selected antimicrobial agents using the agar dilution (AD) and E-Test (ET) methods. We observed no to low resistance against penicillin, ampicillin-sulbactam, meropenem, clindamycin, linezolid and tigecycline (0-2% ET, 0% AD) but high levels of resistance to moxifloxacin (93% ET, 87% AD) and daptomycin (83% ET, 95% AD). The essential agreement of the two methods was very good for benzylpenicillin (EA 95%) and meropenem (EA 92%). The ET method was reliable for correctly categorizing susceptibility, in comparison with the reference method agar dilution, except for daptomycin (categorical agreement 87%). Penicillin is still the first-choice antibiotic for therapy of diseases caused by Actinomyces spp.

It has been shown from the isolation and characterization of exosomes from cell culture media supplemented with fetal bovine serum that both their quality and purity are affected. The high abundance of serum proteins, including bovine cell derived exosomes, is also a potential source of contaminants, which may result in appreciable yields of impure exosomes, thereby leading to artifacts. Isolation and characterization of exosomes from cells maintained under serum‑free conditions should therefore ensure the high quality necessary for medical applications. To meet this end, the present study aimed to characterize exosomes released from THP‑1 macrophages cultured in serum‑free, ultra‑centrifuged medium upon infection with the human pathogen Mycobacterium tuberculosis  (Mtb). Macrophages differentiated from the human cell line THP‑1 were infected at a multiplicity of infection (MOI) of 5. Macrophages were cultivated in CellGenix ® GMP DC serum‑free ultra‑centrifuged medium for 4, 24 and 48 h at 37˚C in a humidified atmosphere with 5% CO 2 . Total exosome isolation reagent was used to extract the exosomes from the cell culture supernatants of naïve and Mtb ‑infected THP‑1 macrophages. The size and purity of the exosomes isolated were subsequently assessed by various methods, including nanoparticle tracking analysis, flow cytometry, MACSPlex exosome analysis, and western blotting. The serum‑free, ultra‑centrifuged medium was found to support the proliferation of the THP‑1 cells successfully. The nanoparticle tracking analysis data revealed that the majority of the isolated particles were within the size range of exosomes (i.e., 30‑150 nM). The MACSPlex exosome analysis confirmed the expression of the exosomal markers, CD9, CD63 and CD81. Furthermore, western blot analysis of the isolated exosomes indicated the presence of CD9, CD63, CD81 and lysosomal associated membrane protein‑1 (LAMP‑1), and also confirmed the absence of Mtb  proteins. Taken together, these data provide evidence that serum‑free, ultra‑centrifuged CellGenix ® GMP DC medium is suitable for application in exosome research, and may significantly advance such studies. Therefore, the use of serum‑free medium for exosome isolation purposes could offer considerable advantages, and constitute a significant improvement in the growing field of extracellular vesicle research. The use of more sensitive methods represents an advance that will enable researchers to rule out the presence of Mtb  pathogenic proteins in exosomes isolated from infected serum‑free cell cultures.

Objectives: Bacteroides spp. are normal constituents of the human intestinal microflora, but they are also able to cause severe diseases. The aim of this study was to determine the diversity of antibiotic resistance genes found in phenotypically resistant Bacteroides and Parabacteroides strains., Methods: A total of 71 phenotypically resistant Bacteroides spp. from human clinical specimens were screened for the antibiotic resistance genes cfiA, tetQ, tetM, tet36, cepA, cfxA, nim, ermG, ermF, bexA, bla VIM , bla NDM , bla KPC , bla OXA-48 and bla GES . The presence of these genes was compared with phenotypic resistance to ampicillin/sulbactam, cefoxitin, ceftolozane/tazobactam, piperacillin/tazobactam, imipenem, meropenem, meropenem/vaborbactam, clindamycin, moxifloxacin, tigecycline, eravacycline and metronidazole., Results: tetQ was the most frequently detected gene, followed by cfiA, ermF, cfxA, ermG, cepA, nim and bexA. None of the strains were positive for tetM, tet36, bla VIM , bla NDM , bla KPC , bla OXA-48 or bla GES . Resistance to the tested β-lactams was mainly linked to the presence of the cfiA gene. Clindamycin resistance correlated with the presence of the genes ermG and ermF. The bexA gene was found in six strains, but only two of them were resistant to moxifloxacin. Tigecycline and eravacycline showed good activities despite the frequent occurrence of tetQ. The nim gene was detected in six isolates, five of which were resistant to metronidazole., Conclusion: The findings of our study support the general belief that antimicrobial resistance within Bacteroides should be taken into consideration. This underlines the necessity of reliable routine antimicrobial susceptibility test methods for anaerobic bacteria and the implementation of antimicrobial surveillance programmes worldwide., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Purpose: The aim of this study was to verify how the prevalence of viridans-streptococci is changed by two appointments of professional prophylaxis and after the subgingival instrumentation via scaling and root planing (SRP). Material and Methods: Samples of the subgingival biofilm were collected from 19 individuals with periodontitis receiving two appointments of professional prophylaxis and SRP before and after the treatment procedures and the presence of viridans-streptococci was analysed by microbiological cultivation. Non-parametric statistical testing using Friedman/Wilcoxon tests and chi-square testing was used for statistical analysis. Results: No statistically significant changes over time were found for the mutans-group. The prevalence of Streptococcus mitis decreased after two appointments of professional prophylaxis (p = 0.013). The prevalence of S. mitis decreased again after SRP (p <0.001). The prevalence of Streptococcus anginosus decreased after two appointments of professional prophylaxis (p = 0.002). After SRP five positive results for S. anginosus were detected (p = 0.026). For Streptococcus oralis and Streptococcus gordonii tendencies to statistical significance were found. The number of positive results for S. oralis increased after the first appointment of professional oral prophylaxis (p = 0.055). The number of positive results for S. gordonii increased after the first appointment of professional oral prophylaxis (p = 0.055). Conclusion: The step-wise periodontal therapy influences the prevalence of viridans-streptococci, especially S. mitis and S. anginosus. No tremendous increase of streptococci especially related to the carious process occurs in the subgingival biofilm. Clinical Relevance: The study reveals knowledge on changes of the composition of the subgingival biofilm due to different steps of periodontal therapy.

Tuberculosis (TB) remains a major health issue worldwide. In order to contain TB infections, improved vaccines as well as accurate and reliable diagnostic tools are desirable. Exosomes are employed for the diagnosis of various diseases. At present, research on exosomes in TB is still at the preliminary stage. Recent studies have described isolation and characterization of Mycobacterium tuberculosis ( Mtb ) derived exosomes in vivo and in vitro. Mtb -derived exosomes ( Mtb exo) may be critical for TB pathogenesis by delivering mycobacterial-derived components to the recipient cells. Proteomic and transcriptomic analysis of Mtb exo have revealed a variety of proteins and miRNA, which are utilized by the TB bacteria for pathogenesis. Exosomes has been isolated in body fluids, are amenable for fast detection, and could contribute as diagnostic or prognostic biomarker to disease control. Extraction of exosomes from biological fluids is essential for the exosome research and requires careful standardization for TB. In this review, we summarized the different studies on Mtb exo molecules, including protein and miRNA and the method used to detect exosomes in biological fluids and cell culture supernatants. Thus, the detection of Mtb exo molecules in biological fluids may have a potential to expedite the diagnosis of TB infection. Moreover, the analysis of Mtb exo may generate new aspects in vaccine development.

Background : Due to the increasing emergence of multi-resistant bacteria the search for alternative antimicrobial substances is of high interest. Promising agents are antimicrobial peptides which are host defense molecules of the innate immune system in a wide range of different species. Objectives : The aim of this study was to assess the activity of nisin, melittin, lactoferrin, parasin-1 and LL-37 against 35 oral bacteria and Candida albicans employing the gold standard method for anaerobic susceptibility testing. Methods : The activity of the peptides was determined by an agar dilution method under anaerobic and aerobic conditions. The test media contained final peptide concentrations between 0.125 µg/ml and 8 µg/ml (melittin, lactoferrin, parasin-1, LL-37) and between 0.125 µg/ml and 128 µg/ml (nisin). Results : Nisin completely inhibited the growth of Megasphaera sp., Bifidobacterium longum, Parvimonas micra, Actinomyces israelii, Actinomyces naeslundii, Actinomyces odontolyticus, Prevotella intermedia, Streptococcus anginosus, Streptococcus constellatus and Staphylococcus aureus . Melittin and lactoferrin reduced the growth of Megasphaera sp., P. micra, B. longum (melittin) and Selenomonas flueggei (lactoferrin). Parasin-1 and LL-37 showed no activity. Conclusion : AMPs, especially nisin and to a smaller degree lactoferrin, might be promising alternatives to antibiotics because of their antimicrobial activity, high resistance to environmental conditions and partially low costs., Competing Interests: The authors report no conflict of interest., (© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Background: While ubiquitously multi-resistant bacteria are on the rise, peri-operative antimicrobial prophylaxis in trauma and near-to-bone surgical procedures has only been changed slightly during the last 25 years. Recent clinical studies concerning the bacterial spectrum and efficacy of antimicrobial treatment in infected trauma surgical patients are rare. The aim of the study was analysis of the contemporary bacterial spectrum and its antimicrobial resistance including the assessment of the appropriateness of peri-operative antimicrobial prophylaxis with cefuroxime. Methods: Patients of a level-I academic trauma center who underwent open or arthroscopic surgery because of a recent trauma necessitating the use of bone-near metal implants were included in the study. All patients in whom a surgical site infection (SSI) had developed during six weeks post-operatively necessitating surgical debridement and had positive microbiologic culture results from an intra-operative surgical site swab were analyzed, retrospectively. In particular, age, gender, date and duration of surgical interventions, and patient's related risk factors were collected, and infecting agents and their minimum inhibitory concentration values for 34 selected antimicrobial agents were evaluated. An SSI occurring later than 6 weeks post-operatively and patients with chronic and septic wounds were excluded. Statistical analysis was performed with SPSS ® (IBM, Armonk, NY). Results: There were 438 pathogens cultured in specimens from 303 enrolled patients (female = 140, male = 163). The most frequent pathogens were Staphylococcus aureus (27.1%), S. epidermidis (20.6%), Enterococcus faecalis (13.6%), Escherichia coli (5.1%), and Pseudomonas aeruginosa (3.7%) accounting for 303 isolates. Of those, 89 (29.4%) were multi-resistant. Of the S. epidermidis isolates, 79.8% (n = 71) were resistant against Oxacillin and thus against most beta-lactam antibiotic agents. Altogether, only 44.1% of the infecting organisms were susceptible to cefuroxime, the antimicrobial agent most often being used for prophylactic purposes. Conclusion: Standardized antimicrobial prophylaxis with cephalosporins has to be reconsidered critically. Multi-resistant species such as S. epidermidis are an increasing challenge in trauma operations.

We analyzed Clostridioides difficile infection (CDI) rates and various antimicrobials' application densities from 2013 to 2019 at Leipzig University Hospital, Germany, by using multivariate linear regression. Ceftriaxone application was the only independent predictor of CDI incidence. Thus, antibiotics' specific pharmacokinetic and pharmacodynamic properties such as biliary excretion of ceftriaxone in its active form should be considered when determining their potential to cause CDI., (© The Author(s) 2020. Published by Oxford University Press on behalf of Infectious Diseases Society of America.)

Objective: Analyze the changes in antimicrobial drug resistance patterns due to hospital relocation. To this end, we conducted a retrospective analysis of microbiological results, especially minimum inhibitory concentrations (MIC) of all isolates from blood, urine and bronchial secretions, in our pediatric university hospital before and after moving to a new building., Results: While the number of tests done did not change, the fraction of those positive increased, more MICs were determined and certain microbes (A. baumannii, E. faecalis, Klebsiella spp. and P. mirabilis) were detected more frequently. Most changes in MICs occurred in E. faecium (increases in 8 antimicrobials, decreases only in linezolid and gentamicin). For imipenem and aminopenicillins the MICs commonly rose after relocation, the opposite is true for gentamicin and trimethoprim/sulfamethoxazole. The other factors that alter by moving a hospital such as changes in medical personnel or case severity cannot be corrected for, but using MICs we are able to provide insights into changes down to the individual antimicrobial drug and even small changes usually undetectable to the common categorical reporting of resistance.

Background: Mediastinitis after cardiac surgery can lead to devastating consequences such as deep sternal wound infections (DSWI). Staphylococcus epidermidis and other coagulase-negative staphylococci belong to the physiological skin flora and therefore generally are not considered pathogenic agents. Thus, local resistance patterns of these bacterial species often recovered from wound specimens generally are ignored while choosing antibiotics for peri-operative prophylaxis in cardiac surgery as well as in the selection of empiric antibiotic therapy of DSWI. Methods: During the period May 2012-May 2013, 52 patients suffering from DSWI were treated at our institution. For every patient, deep tissue samples were obtained during surgical debridement procedures and submitted to microbiologic analysis. The frequency of and the time to occurrence of a DSWI was recorded, and baseline data, previous operative interventions, complications, and the technique used for soft tissue reconstruction, as well as the microbiologic results and individual risk factors, were documented. Results: There were 32 male patients (62%) and 20 female. The patients' age at the time of revision was a mean of 67 ± 11.5 years (range 35-83 years). There was bacterial growth in 31 cases (60%), the predominant species being S. epidermidis (20 patients; 65%). Extended antibiotic therapy was indispensable to controlling the infection. Conclusion: The local resistance patterns of antibiotics should have a greater influence on the standardized prophylaxis or empirical therapy of DSWI and need to be discussed specifically for this high-risk population. Because of its multi-resistance spectrum, S. epidermidis must be classified as a potential pathogen. In the cases reported here, extended antibiotic therapy was necessary to support wound healing and thus good patient outcomes.

This is the seventh chapter of the guideline "Calculated initial parenteral treatment of bacterial infections in adults - update 2018" in the 2 nd updated version. The German guideline by the Paul-Ehrlich-Gesellschaft für Chemotherapie e.V. (PEG) has been translated to address an international audience. The chapter deals with the empirical and targeted antimicrobial therapy of complicated intra-abdominal infections. It includes recommendations for antibacterial and antifungal treatment., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2020 Eckmann et al.)

Background: Intercontinental travel contributes to the spread of extended-spectrum beta-lactamase producing Enterobacterales (ESBL-PE). We assessed risk factors for intestinal ESBL-PE colonization in people travelling to low and middle income countries in the tropics and subtropics to better understand how travel affects ESBL-PE spread., Method: This prospective cohort study in travellers attending a travel clinic in Leipzig, Germany was conducted in 2016-2017. Information on risk factors related to travel, symptoms, antibiotic use, health care usage, accommodation, destination, diet and hygiene was collected by questionnaire after travel. Stools were phenotypically tested for ESBL-PE before and after travel. Risk factors for ESBL-PE colonization were identified using logistic regression., Results: Of the 230 travellers that were ESBL-PE negative before travelling, 23% (n = 53) travellers returned positive. Multivariable analyses showed that age, type of accommodation and travelling to Asia were associated with ESBL-PE colonization., Conclusions: Given that a considerable amount of travellers returned with ESBL-PE, we recommend raising awareness in returning high-risk travellers, e.g. those returning from high-risk areas. They should be aware that they may carry antimicrobial-resistant bacteria after travel, and how they can prevent its spread. The role of the type of accommodation as a factor favouring intestinal colonization with ESBL-PE requires further investigation., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Multidrug-resistant gram-negative bacteria (MDR-GNB) constitute a threat to health care worldwide. Disinfectants are used to prevent and control the spread of MDR-GNB in a hospital setting but their efficacy might be impaired by bacterial mechanisms that may act on both antimicrobials and disinfectants. Determination of minimum inhibitory concentrations is mainly used to determine bacterial susceptibility against disinfectants, but practical tests on surfaces might be more suitable to predict in-use conditions. Our objective was to compare and evaluate 4 different methods widely used to assess surface disinfectant efficacy., Methods: The efficacy of benzalkonium chloride (BAC), peracetic acid (PAA), and ethanol (ETH) against multidrug-resistant Acinetobacter, Pseudomonas, and Klebsiella strains was assessed by minimum inhibitory concentration determinations, quantitative suspension tests, qualitative suspension tests, and carrier tests. Test results were compared to ascertain the most appropriate method., Results: ETH, PAA, and BAC were highly effective against MDR-GNB, but we observed marked differences in efficacious concentrations (up to 100-fold) as a function of the test method applied. Minimum inhibitory concentration determination was not reliable for evaluating susceptibility or resistance to BAC., Conclusions: Surface tests should be used to determine bacterial susceptibility against disinfectants. Moreover, suitable guidelines are needed that allow for the standardization and comparison of bactericidal values obtained by different investigators., (Copyright © 2019 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Objective: The aim of this study was to investigate the characteristics of bacteremia caused by professional mechanical plaque removal (PMPR) in two groups of patients with generalized moderate chronic periodontitis., Materials and Methods: Venous blood samples were taken at multiple time points for one hour following PMPR in fifty patients with generalized moderate chronic periodontitis. Subjects consisted of two groups, one group was receiving supportive periodontal therapy (SPT, n = 25) and the other group was receiving initial periodontal therapy (IPT, n = 25). Blood samples were processed and analyzed for cultivable microflora. Pertinent clinical parameters were recorded for each patient in both groups., Results: Bacteremia was detected in 10 of 25 SPT and 8 of 25 IPT patients (p = 0.796). In both groups, the prevalence of bacteremia was dependent on the time of blood sampling and varied in magnitude between <10 2  CFU/ml and 10 6  CFU/ml. Sixteen different bacterial species were identified in both groups, mostly Actinomyces naeslundii (SPT n = 3, IPT n = 4) and Streptococcus spp. (SPT n = 6, IPT n = 2). In regression models, Grade II furcation involvement (p = 0.004) and Gingival Bleeding Index (p = 0.036) had affected the occurrence of bacteremia but in the SPT group only., Conclusion: Professional mechanical plaque removal was associated with bacteremia regardless of whether a patient was receiving SPT or IPT., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.)

Background: Symptomatic infection with pinworm (Enterobius vermicularis), a human pathogen, is clinically relevant in Germany, with an estimated prevalence in childhood of 2-20%. Enterobiasis can cause major mental distress. There is little systematically verified knowledge on the treatment of this condition, and there is no corresponding German guideline. This review is, therefore, intended as a summary of the current state of knowledge., Methods: This review is based on pertinent publications retrieved by a selective search in PubMed for literature appearing from 1 January 1990 to 5 February 2019 and containing the search terms "enterobiasis," "oxyuriasis," "Enterobius vermicula- ris," "pinworm," and "threadworm.", Results: More than one billion people worldwide are thought to be infected with pinworm. Estimates of its prevalence among kindergarten and primary-school pupils in Europe are generally near 20%. Infants (<2 years of age), adolescents (>14 years of age), and adults are only sporadically affected. The main risk factors are age 4-11 years, uncontrolled anus-finger-mouth contact, nail-biting (onychophagia/peri- onychophagia), unsupervised body hygiene, and poor compliance with basic hand hygiene. No large-scale, randomized, controlled trials of treatment are available. The approved antihelminthic agents are mebendazole, pyrantel embonate, and pyrvinium embonate (success rates up to >90%). For recurrent infections, prolonged treatment for up to 16 weeks (a "pulse scheme") is recommended., Conclusion: In nearly all cases, antihelminthic treatment along with attention to hygienic measures can successfully eradicate pinworm infection and prevent recurrence and autoinfection. The involvement of all persons living in the patient's house- hold, including sexual partners, is a prerequisite to the lasting success of treatment.

The genus Clostridium is of high clinical relevance, as some species may cause rapid and even lethal infections. Thus, a timely identification of these anaerobic bacteria is desirable. Conventional identification methods rely on biochemical properties of these organisms, however, establishing these is time-consuming and not always reliable. Alternatively, 16S rRNA gene sequence based diagnostic methods may be used, but they are expensive and not ubiquitously available. This study was designed to assess the possibility to identify Clostridium species employing the matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). For this purpose, 848 Clostridium strains representing 42 species were analyzed with the VITEK ® MS instrument (bioMérieux, Marcy l'Etoile, France), comparing mass spectra derived from these organisms with the spectra provided in the available database. 90.3% of the strains were correctly identified at species level and another 3.6% at genus level. Since the number of Clostridium species included in the database was rather limited (21 altogether), the spectra obtained were also analyzed employing the Shimadzu Pro Series software. Thus, it became possible to create a dendrogram of the species included in this study., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Clostridium species cause several local and systemic diseases. Conventional identification of these microorganisms is in part laborious, not always reliable, time consuming or does not always distinguish different species, i.e., C. botulinum and C. sporogenes. All in, there is a high interest to find out a reliable, powerful and rapid method to identify Clostridium spp. not only on genus but also on species level. The aim of the present study was to identify Clostridium spp. strains and also to find differences and metabolic groups of C. botulinum by Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS). A total of 123 strains of Clostridium spp. (C. botulinum, n = 40, C. difficile, n = 11, C. tetani, n = 11, C. sordellii, n = 20, C. sporogenes, n = 18, C. innocuum, n = 10, C. perfringens, n = 13) were analyzed by MALDI-TOF MS in combination with methods of multivariate statistical analysis. MALDI-TOF MS analysis in combination with methods of multivariate statistical analysis was able to discriminate between the different tested Clostridium spp., even between species which are closely related and difficult to differentiate by traditional methods, i.e., C. sporogenes and C. botulinum. Furthermore, the method was able to separate the different metabolic groups of C. botulinum. Especially, E gene-positive C. botulinum strains are clearly distinguishable from the other species but also from those producing other toxin types. Thus, MALDI-TOF MS represents a reliable and above all quick method for identification of cultivated Clostridium species.

Background: Increased antimicrobial resistance has been observed among many bacteria leading to treatment failures in human and veterinary medicine. Disinfection is a prerequisite for infection control and prevention in healthcare settings. Chlorine compounds are cost-effective and accessible worldwide., Aim: To determine the efficacy of sodium hypochlorite (NaOCl) against multidrug-resistant Gram-negative bacteria (MDR-GNB)., Methods: Minimum inhibitory concentrations (MICs) were determined using broth macro-dilution. Bactericidal efficacy was measured by qualitative and quantitative suspension tests followed by practical tests without mechanical action on stainless steel carriers. The guidelines of the German Association for Applied Hygiene were followed., Findings: Results varied remarkably depending on the method. MICs were 0.1% or 0.2% NaOCl. Qualitative suspension tests revealed up to 500-fold lower bactericidal concentrations. Pseudomonas aeruginosa (P = 0.0025) was significantly less susceptible in these tests whereas quantitative suspension tests revealed no significant differences between strains (P > 0.05). Practical tests determined bactericidal concentrations of 0.8-0.32% NaOCl at 1 min of contact and even lower concentrations for longer contact times. At 1 min, five Klebsiella were significantly less susceptible (P = 0.0124), whereas the lower susceptibility of P. aeruginosa was not confirmed. Organic load inhibited bactericidal activity significantly, whereas contact time had a marginal effect. Differing test results underline that MIC determination and qualitative suspension tests may be insufficient approaches to evaluate bacterial susceptibility or resistance., Conclusion: NaOCl efficiently reduced Pseudomonas aeruginosa, Acinetobacter spp., and Klebsiella spp., most notably in the absence of organic matter. Strain- and species-specific differences in susceptibility were noticed, but in general MDR-GNB revealed no higher tolerance to NaOCl., (Copyright © 2018 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

In this study, the effect of bacterial multispecies communities on the adhesion of Streptococcus mutans and Streptococcus sanguinis to dental restorative material was investigated. The saliva-coated specimens of zirconia and composite were incubated with the following combinations: single species, S. mutans or S. sanguinis ; two species, single species combined with other oral streptococci; multiple species, combination of Actinomyces naeslundii , Fusobacterium nucleatum , and Prevotella ssp.; and the two-species combinations. The adherent bacteria were counted after plating of serial dilutions. Effects of material and bacteria on adhesion of S. mutans and S. sanguinis were evaluated with multiple linear regression analyses. No significant differences between the materials regarding the adhesion of S. mutans and S. sanguinis were observed. The adhesion of S. mutans was negatively influenced by the presence of other streptococci. Enhancing effects (610.6%) were seen in the presence of Prevotella intermedia . The adhesion of S. sanguinis decreased in the presence of other bacteria, except F. nucleatum (increase of 717.4%). Significant inhibitory effects were detected in the presence of S. mutans and A. naeslundii (reduction of 95.9% and 78.5%, respectively). The results of this study suggest that adhesion of both types of streptococci to restorative materials is influenced by various bacterial interactions.

The Tigecycline Evaluation and Surveillance Trial (T.E.S.T.) is a global program that aims to monitor the in vitro antimicrobial activities of current therapeutic agents against clinical isolates. This study presents surveillance data for Gram-positive and Gram-negative anaerobic isolates (N = 7008) collected from nine European countries between 2010 and 2016. Presented in this study are antimicrobial susceptibility data, according to the European Committee for Antimicrobial Susceptibility Testing (EUCAST) breakpoints, and minimum inhibitory concentration (MIC) distributions. The antimicrobial agents tested were cefoxitin (Gram-negative isolates only), clindamycin, meropenem, metronidazole, penicillin (Gram-positive isolates only), piperacillin-tazobactam and tigecycline. Among all Gram-positive and Gram-negative anaerobes, the lowest rates of resistance were to meropenem and metronidazole (0.0%-1.7% and 0.0%-1.9%, respectively). High rates of resistance were reported to clindamycin, in particular among isolates of the Bacteroides fragilis group (22.1%-48.1%) and Prevotella spp. (10.9%-32.2%). The majority of MIC distributions were unimodal, with the exception of clindamycin, which were mostly bimodal. Fifty percent of Gram-negative isolates gave tigecycline MICs between 0.06 and 1 mg/L, and 50% of Gram-positive isolates exhibited tigecycline MICs between 0.06 and 0.25 mg/L. The findings of this study suggest that the majority of anaerobic isolates were susceptible to meropenem and metronidazole, and that tigecycline remained active, but clindamycin resistance is a cause for concern in Europe. Surveillance studies, such as T.E.S.T., provide information on changes in the susceptibility of clinically important pathogens to commonly prescribed antimicrobial agents, and can highlight problems of antimicrobial resistance that need to be addressed., (Copyright © 2018 Pfizer Inc. Published by Elsevier Ltd.. All rights reserved.)

Background: From July 2010-April 2013, Leipzig University Hospital experienced the largest outbreak of a Klebsiella pneumoniae carbapenemase 2 (KPC-2)-producing Klebsiella pneumoniae (KPC-2-Kp) strain observed in Germany to date. After termination of the outbreak, we aimed to reconstruct transmission pathways by phylogenetics based on whole-genome sequencing (WGS)., Methods: One hundred seventeen KPC-2-Kp isolates from 89 outbreak patients, 5 environmental KPC-2-Kp isolates, and 24 K pneumoniae strains not linked to the outbreak underwent WGS. Phylogenetic analysis was performed blinded to clinical data and based on the genomic reads., Results: A patient from Greece was confirmed as the source of the outbreak. Transmission pathways for 11 out of 89 patients (12.4%) were plausibly explained by descriptive epidemiology, applying strict definitions. Five of these and an additional 15 (ie, 20 out of 89 patients [22.5%]) were confirmed by phylogenetics. The rate of phylogenetically confirmed transmissions increased significantly from 8 out of 66 (12.1% for the time period before) to 12 out of 23 patients (52.2% for the time period after; P <.001) after implementation of systematic screening for KPC-2-Kp (33,623 screening investigations within 11 months). Using descriptive epidemiology, systematic screening showed no significant effect (7 out of 66 [10.6%] vs 4 out of 23 [17.4%] patients; P = .465). The phylogenetic analysis supported the assumption that a contaminated positioning pillow served as a reservoir for the persistence of KPC-2-Kp., Conclusions: Effective phylogenetic identification of transmissions requires systematic microbiologic screening. Extensive screening and phylogenetic analysis based on WGS should be started as soon as possible in a bacterial outbreak situation., (Copyright © 2018 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Background: In 2016, an increased rate of methicillin-susceptible Staphylococcus aureus colonization was detected on a neonatal intensive care unit at the Leipzig University Hospital. Typing results showed a predominant spa -type t091. Considering nosocomial clustering, several infection prevention measures (e. g. intensified standard precautions, single-occupancy room, cohorted patients, continuing education of staff) were introduced, including staff screening followed by decolonization of colonized health care workers., Methods: Staff members showing positive on screening carried out a 5-day decolonization program at home. Decolonization products containing octenidine as active ingredient were used first. At the earliest, 48 h after completing the procedure, the success of the intervention was tested (3 buccal and nasal swabs were taken on consecutive days). If 2 attempts at decolonization were not successful, staff members were provided with a mupirocin-containing nasal ointment instead of octenidine., Results: Of 128 employees examined, 43 (33.6%) were identified as carriers of S. aureus . In 9 cases (20.9%; 9/43) the S. aureus matched with type t091. 2 carriers (4.7%; 2/43) of MRSA were detected as well. The first decolonization attempt against t091 and MRSA failed altogether. After a second decolonization, 3 cases became negative. Finally, 8 remaining staff members were decolonized successfully with mupirocin containing nasal ointment., Conclusions: Various reasons might explain the difficulties of decolonization such as the challenge of managing decolonization at home, inhibitory factors as well as inconsistent performance of decolonization measures. Additionally, differences between the preparations for the nasal decontamination may be considered., Competing Interests: Die Autoren geben an, dass kein Interessenkonflikt besteht., (© Georg Thieme Verlag KG Stuttgart · New York.)

Two matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based methods were compared for their ability to identify viridans streptococci. One approach employed a reference database and software developed in-house. All inhouse measurements were performed using an Autoflex II Instrument (Bruker Daltonics GmbH, Germany). The other system, a VITEK-MS (BioMérieux, France) was operated on the commercially available V2.0 Knowledge Base for Clinical Use database. Clinical isolates of viridans streptococci (n=184) were examined. Discrepant results were resolved by 16S rDNA sequencing. Species-level identification percentages were compared by a chi-square test. The in-house method correctly identified 179 (97%) and 175 (95%) isolates to the group and species level respectively. In comparison, the VITEK-MS system correctly identified 145 (79%) isolates to the group and species level. The difference between the two methods was statistically significant at both group and species levels. Using the Autoflex II instrument combined with an extraction method instead of whole cell analysis resulted in more reliable viridans streptococci identification. Our results suggest that combining extraction with powerful analysis software and the careful choice of well-identified strains included into the database was useful for identifying viridans streptococci species., Competing Interests: No potential conflicts of interest relevant to this article were reported., (© The Korean Society for Laboratory Medicine.)

Purpose: High antibiotic and antifungal concentrations in wastewater from anti-infective drug production may exert selection pressure for multidrug-resistant (MDR) pathogens. We investigated the environmental presence of active pharmaceutical ingredients and their association with MDR Gram-negative bacteria in Hyderabad, South India, a major production area for the global bulk drug market., Methods: From Nov 19 to 28, 2016, water samples were collected from the direct environment of bulk drug manufacturing facilities, the vicinity of two sewage treatment plants, the Musi River, and habitats in Hyderabad and nearby villages. Samples were analyzed for 25 anti-infective pharmaceuticals with liquid chromatography-tandem mass spectrometry and for MDR Gram-negative bacteria using chromogenic culture media. In addition, specimens were screened with PCR for bla VIM , bla KPC , bla NDM , bla IMP-1 , and bla OXA-48 resistance genes., Results: All environmental specimens from 28 different sampling sites were contaminated with antimicrobials. High concentrations of moxifloxacin, voriconazole, and fluconazole (up to 694.1, 2500, and 236,950 µg/L, respectively) as well as increased concentrations of eight other antibiotics were found in sewers in the Patancheru-Bollaram industrial area. Corresponding microbiological analyses revealed an extensive presence of extended-spectrum beta-lactamase and carbapenemase-producing Enterobacteriaceae and non-fermenters (carrying mainly bla OXA-48 , bla NDM , and bla KPC ) in more than 95% of the samples., Conclusions: Insufficient wastewater management by bulk drug manufacturing facilities leads to unprecedented contamination of water resources with antimicrobial pharmaceuticals, which seems to be associated with the selection and dissemination of carbapenemase-producing pathogens. The development and global spread of antimicrobial resistance present a major challenge for pharmaceutical producers and regulatory agencies.

Aims: In infective endocarditis (IE), a severe inflammatory disease of the endocardium with an unchanged incidence and mortality rate over the past decades, only 1% of the cases have been described as polymicrobial infections based on microbiological approaches. The aim of this study was to identify potential biodiversity of bacterial species from infected native and prosthetic valves. Furthermore, we compared the ultrastructural micro-environments to detect the localization and distribution patterns of pathogens in IE., Material and Methods: Using next-generation sequencing (NGS) of 16S rDNA, which allows analysis of the entire bacterial community within a single sample, we investigated the biodiversity of infectious bacterial species from resected native and prosthetic valves in a clinical cohort of 8 IE patients. Furthermore, we investigated the ultrastructural infected valve micro-environment by focused ion beam scanning electron microscopy (FIB-SEM)., Results: Biodiversity was detected in 7 of 8 resected heart valves. This comprised 13 bacterial genera and 16 species. In addition to 11 pathogens already described as being IE related, 5 bacterial species were identified as having a novel association. In contrast, valve and blood culture-based diagnosis revealed only 4 species from 3 bacterial genera and did not show any relevant antibiotic resistance. The antibiotics chosen on this basis for treatment, however, did not cover the bacterial spectra identified by our amplicon sequencing analysis in 4 of 8 cases. In addition to intramural distribution patterns of infective bacteria, intracellular localization with evidence of bacterial immune escape mechanisms was identified., Conclusion: The high frequency of polymicrobial infections, pathogen diversity, and intracellular persistence of common IE-causing bacteria may provide clues to help explain the persistent and devastating mortality rate observed for IE. Improved bacterial diagnosis by 16S rDNA NGS that increases the ability to tailor antibiotic therapy may result in improved outcomes.

Background: European centers (n = 226) involved in the Tigecycline Evaluation and Surveillance Trial (TEST, 2004-2014) submitted data and bacterial isolates., Methods: Minimal inhibitory concentrations and susceptibility were determined using Clinical and Laboratory Standards Institute methods and European Committee on Antimicrobial Susceptibility Testing breakpoints., Results: The rates of the following resistant pathogens increased from 2004 to 2014: extended-spectrum β-lactamase (ESBL)-positive Escherichia coli (from 8.9 to 16.9%), multidrug-resistant Acinetobacter baumannii complex (from 15.4 to 48.5%), and ESBL-positive Klebsiella pneumoniae (from 17.2 to 23.7%). The rate of methicillin-resistant Staphylococcus aureus was 27.5% in 2004 and 28.9% in 2014. Resistance to the carbapenems (imipenem and meropenem) was 37.4 and 14.5% for A. baumannii complex and Pseudomonas aeruginosa, respectively. Carbapenem resistance was ≤4.3% among the Enterobacteriaceae and 0.2% against Streptococcus pneumoniae. The resistance to tigecycline ranged between 7.4% against ESBL-producing K. pneumoniae and 0.0% against S. aureus., Conclusions: The carbapenems and tigecycline were active against Enterobacteriaceae. Agents with activity against A. baumannii complex and P. aeruginosa are limited. The carbapenems, tigecycline, linezolid, and vancomycin were active against Gram-positive organisms., (© 2016 S. Karger AG, Basel.)

Introduction: Clostridium difficile infection (CDI) is the most common cause of health-care-associated infectious diarrhea. Recurrence rates are as high as 20-30% after standard treatment with metronidazole or vancomycin, and appear to be reduced for patients treated with fidaxomicin. According to the literature, the risk of CDI recurrence increases after the second relapse to 30-65%. Accurate data for Germany are not yet available., Methods: Based on the research database of arvato health analytics (Munich, Germany), a secondary data analysis for the incidence, treatment characteristics and course of CDI was performed. The database included high granular accounting information of about 1.46 million medically insured patients covering the period 2006-2013, being representative for Germany. The analysis was based on new-onset CDI in 2012 in patients which either received outpatient antibiotic therapy for CDI or were hospitalized., Results: The ICD-10 coded incidence of CDI in 2012 was 83 cases per 100,000 population. Overall mortality rates within the follow-up period of 1 year were 13.5% in inpatients with primary diagnosis of CDI, compared to 24.3% in inpatients with secondary diagnosis of CDI (P < 0.001), and 7.1% in outpatients (P < 0.001). In the median, patients with secondary diagnosis of CDI remained significantly longer hospitalized (24 vs. 9 days, P < 0.001). First recurrence of CDI was observed in 18.2% of cases with index events. There was a significantly increased risk to suffer a second and third recurrence, reaching 28.4% (P < 0.001), and 30.2% (P = 0.017), respectively. Antibiotic therapy of CDI in outpatients was performed mainly with metronidazole (in 90.8% of index events, 60.0% of first recurrences, and 43.5% of second recurrences)., Conclusion: The reported incidence of CDI in Germany is higher than noted previously. The recurrence rates do increase with the number of relapses, but are lower than reported in the literature, despite dominance of metronidazole treatment in outpatients., Funding: MSD Sharp & Dohme GmbH, Haar, Germany.

Background: Plastic stents used for the treatment of biliary obstruction will become occluded over time due to microbial colonization and formation of biofilms. Treatment of stent-associated cholangitis is often not effective because of inappropriate use of antimicrobial agents or antimicrobial resistance. We aimed to assess the current bacterial and fungal etiology of stent-associated biofilms, with particular emphasis on antimicrobial resistance., Methods: Patients with biliary strictures requiring endoscopic stent placement were prospectively enrolled. After the retrieval of stents, biofilms were disrupted by sonication, microorganisms were cultured, and isolates were identified by matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and/or biochemical typing. Finally, minimum inhibitory concentrations (MICs) were determined for various antimicrobial agents. Selected stents were further analyzed by fluorescence in situ hybridization (FISH)., Results: Among 120 patients (62.5% males, median age 64 years) with biliary strictures (35% malignant, 65% benign), 113 double pigtail polyurethane and 100 straight polyethylene stents were analyzed after a median indwelling time of 63 days (range, 1-1274 days). The stent occlusion rate was 11.5% and 13%, respectively, being associated with a significantly increased risk of cholangitis (38.5% vs. 9.1%, P<0.001). Ninety-five different bacterial and 13 fungal species were detected; polymicrobial colonization predominated (95.8% vs. 4.2%, P<0.001). Enterococci (79.3%), Enterobacteriaceae (73.7%), and Candida spp. (55.9%) were the leading pathogens. Candida species were more frequent in patients previously receiving prolonged antibiotic therapy (63% vs. 46.7%, P = 0.023). Vancomycin-resistant enterococci accounted for 13.7%, extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae with co-resistance to ciprofloxacin accounted for 13.9%, and azole-resistant Candida spp. accounted for 32.9% of the respective isolates., Conclusions: Enterococci and Candida species play an important role in the microbial colonization of biliary stents. Therefore, empirical antimicrobial treatment of stent-associated cholangitis should be guided toward enterococci, Enterobacteriaceae, streptococci, anaerobes, and Candida. To determine causative pathogens, an accurate microbiological analysis of the extracted stent(s) may be helpful.