Early-Outcome Differences between Acute and Chronic Periprosthetic Joint Infections-A Retrospective Single-Center Study.

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.

Extended-spectrum beta-lactamases found in Escherichia coli isolates obtained from blood cultures and corresponding stool specimen.

With extended-spectrum ß-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.

Leclercia pneumoniae sp. nov., a bacterium isolated from clinical specimen in Leipzig, Germany.

Strain 49125T 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 49125T represents a new species, for which we propose the name Leclercia pneumoniae sp. nov. with the type strain 49125T (=LMG 32245T=DSM 112336T).

Antimicrobial Susceptibility of Clinical Oral Isolates of Actinomyces spp.

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.

Spinal Tuberculosis. / Tuberkulose der Wirbelsäule.

Tuberculosis is one of the most common infectious diseases worldwide. The frequency in Germany is low, however, an increase has been observed in the past few years. The incidence of extrapulmonary manifestation accounts for up to 10 to 20%. In 50% of these cases the spinal column is affected. Although literature reveals worldwide experiences in the treatment, in Germany spinal tuberculosis remains a rarity. Different pitfalls and specific characteristics regarding diagnosis and therapy are to consider. Therefore, a presentation of these specifics and their discussion based on the available literature will be presented. The purpose is to achieve an increase in awareness regarding this, in our latitudes, rare disease.

[Antibiotics Stewardship Team Recommendations - An Analysis under Routine Conditions in a University Hospital]. / Empfehlungen des Antibiotic-Stewardship Teams ­ eine Analyse unter Routinebedingungen eines Universitätsklinikums.

AIM: To evaluate recommendations of an antibiotic stewardship (ABS) team during routine weekly visits and to analyse their implementation and reasons for non-implementation by the ward physicians. METHODS: Multiprofessional ABS Rounds (members: infectiology, microbiology, pharmacy and hospital epidemiology) were accompanied by an observer in nine intensive and peripheral wards of a university hospital (1451 beds) for eight weeks. The recommendations of the ABS team were prospectively analysed, and when these were not implemented, ward physicians in charge were asked to give reasons for non-implementation. RESULTS: A total of 262 patients were visited in the course of 359 ABS visits. A median of four physicians and one pharmacist (Q25/Q75: 4/6) participated in the ABS rounds. In 177/359 (49%) of visits, at least one recommendation for anti-infective therapy was given; the total number of recommendations made was 210. The most frequent (38%, 80/210) recommendations were related to the prescribed therapy duration. The more different the professional groups participating in the ABS rounds, the larger was the number of recommendations given (p=0.016; odds ratio=1.018 (1.003-1.033)). 62/210 (30%) of the ABS recommendations were not implemented by the ward physicians in charge. The most frequent reason (32%: 20/62) for this was the deliberate decision by the physicians to deviate from the recommended therapy. CONCLUSIONS: The high recommendation rate of 50% demonstrates the need for optimizing therapy by the ABS teams in routine practice. The larger the number of different specialists participating in single visits, the larger was the number of recommendations. Reasons for the lack of implementations need to be critically examined by both the ABS team and the ward physicians in charge.

Exosomes in serum­free cultures of THP­1 macrophages infected with Mycobacterium tuberculosis.

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 serum­free conditions should therefore ensure the high quality necessary for medical applications. To meet this end, the present study aimed to characterize exosomes released from THP­1 macrophages cultured in serum­free, ultra­centrifuged medium upon infection with the human pathogen Mycobacterium tuberculosis (Mtb). Macrophages differentiated from the human cell line THP­1 were infected at a multiplicity of infection (MOI) of 5. Macrophages were cultivated in CellGenix® GMP DC serum­free ultra­centrifuged medium for 4, 24 and 48 h at 37˚C in a humidified atmosphere with 5% CO2. Total exosome isolation reagent was used to extract the exosomes from the cell culture supernatants of naïve and Mtb­infected THP­1 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 serum­free, ultra­centrifuged medium was found to support the proliferation of the THP­1 cells successfully. The nanoparticle tracking analysis data revealed that the majority of the isolated particles were within the size range of exosomes (i.e., 30­150 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 protein­1 (LAMP­1), and also confirmed the absence of Mtb proteins. Taken together, these data provide evidence that serum­free, ultra­centrifuged CellGenix® GMP DC medium is suitable for application in exosome research, and may significantly advance such studies. Therefore, the use of serum­free 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 serum­free cell cultures.

Fluorescence in situ Hybridization (FISH) in the Microbiological Diagnostic of Deep Sternal Wound Infection (DSWI).

PURPOSE: 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.

Diversity of antimicrobial resistance genes in Bacteroides and Parabacteroides strains isolated in Germany.

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, blaVIM, blaNDM, blaKPC, blaOXA-48 and blaGES. 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, blaVIM, blaNDM, blaKPC, blaOXA-48 or blaGES. Resistance to the tested ß-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.

Composition and Clinical Significance of Exosomes in Tuberculosis: A Systematic Literature Review.

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 (Mtbexo) may be critical for TB pathogenesis by delivering mycobacterial-derived components to the recipient cells. Proteomic and transcriptomic analysis of Mtbexo 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 Mtbexo molecules, including protein and miRNA and the method used to detect exosomes in biological fluids and cell culture supernatants. Thus, the detection of Mtbexo molecules in biological fluids may have a potential to expedite the diagnosis of TB infection. Moreover, the analysis of Mtbexo may generate new aspects in vaccine development.

Streptococci in the Subgingival Biofilm and Periodontal Therapy.

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.

Activity of five antimicrobial peptides against periodontal as well as non-periodontal pathogenic strains.

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.

Switching From Ceftriaxone to Cefotaxime Significantly Contributes to Reducing the Burden of Clostridioides difficile infections.

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.

Relocating a pediatric hospital: Does antimicrobial resistance change?

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.

Calculated parenteral initial treatment of bacterial infections: Intra-abdominal infections.

This is the seventh chapter of the guideline "Calculated initial parenteral treatment of bacterial infections in adults - update 2018" in the 2nd 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.

Development of the Bacterial Spectrum and Antimicrobial Resistance in Surgical Site Infections of Trauma Patients.

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.

Clinical and Microbiological Analysis of Deep Sternal Wound Infections in Fifty-Two Consecutive Patients.

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.

Intestinal colonization with extended-spectrum beta-lactamase producing Enterobacterales (ESBL-PE) during long distance travel: A cohort study in a German travel clinic (2016-2017).

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.