Draft genome sequences of a blaCTX-M-15-harboring and an atypical enteropathogenic Escherichia coli isolate recovered from slaughterhouses in Nigeria.

We present the draft genome sequences of two Escherichia coli strains isolated from slaughterhouses in Edo State, Nigeria, in 2019. The isolates were identified as blaCTX-M-15-harboring (19-47-58) and atypical enteropathogenic E. coli (aEPEC) (19-47-66), belonging to multilocus sequence types (MLST) ST46 and ST2089, respectively.

Surgical Management of Periprosthetic Joint Infections in Hip and Knee Megaprostheses.

Periprosthetic joint infection is a feared complication after the megaprosthetic reconstruction of oncologic and non-oncologic bone defects of including the knee or hip joint. Due to the relative rarity of these procedures, however, optimal management is debatable. Considering the expanding use of megaprostheses in revision arthroplasty and the high revision burden in orthopedic oncology, the risk of PJI is likely to increase over the coming years. In this non-systematic review article, we present and discuss current management options and the associated results focusing on studies from the last 15 years and studies from dedicated centers or study groups. The indication, surgical details and results in controlling infection are presented for debridement, antibiotics, irrigation and retention (DAIR) procedure with an exchange of the modular components, single-stage implant exchange, two-stage exchanges and ablative procedures.

On the potential role of naturally occurring carboxylic organic acids as anti-infective agents: opportunities and challenges.

Carboxylic organic acids are intermediates of central carbon metabolic pathways (e.g. acetic, propionic, citric, and lactic acid) long known to have potent antimicrobial potential, mainly at acidic pHs. The food industry has been leveraging those properties for years, using many of these acids as preservatives to inhibit the growth of pathogenic and/or spoilage fungal and bacterial species. A few of these molecules (the most prominent being acetic acid) have been used as antiseptics since Hippocratic medicine, mainly to treat infected wounds in patients with burns. With the growth of antibiotic therapy, the use of carboxylic acids (and other chemical antiseptics) in clinical settings lost relevance; however, with the continuous emergence of multi-antibiotic/antifungal resistant strains, the search for alternatives has intensified. This prospective article raises awareness of the potential of carboxylic acids to control infections in clinical settings, considering not only their previous exploitation in this context (which we overview) but also the positive experience of their safe use in food preservation. At a time of great concern with antimicrobial resistance and the slow arrival of new antimicrobial therapeutics to the market, further exploration of organic acids as anti-infective molecules may pave the way to more sustainable prophylactic and therapeutic approaches.

Survey on microbial contamination of opened skin care products used for tattooing.

AIMS: Inadequate hygiene measures as well as the use of contaminated inks or non-sterile needles are considered as important infection sources in the process of tattooing. In tattoo parlors and at conventions, it is common practice to apply cosmetic products from bulk packs as lubricants during tattooing and as ointments for tattoo aftercare. The objective of our study was to assess the microbial load of opened skin care products used during tattooing or for tattoo aftercare. METHODS AND RESULTS: First, we established a homogenization method suitable for the microbiological examination of water-immiscible products. To this end, we compared the efficiency of FastPrepTM and Stomacher® homogenizers on artificially contaminated petroleum jelly. FastPrep homogenates revealed significantly higher detection rates (≥97%) compared to Stomacher ones (31%-64%). Second, we investigated 106 cosmetic bulk pack products collected from tattoo artists. After FastPrep homogenization for 30 seconds, total aerobic mesophilic bacteria and the presence of Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans were determined through culture. We also tested for Mycobacteria spp. by qPCR. In total, 4.7% of the cosmetic products under study turned out to be contaminated. CONCLUSION: The observed microbial contamination of opened skin care bulk packs can hold a risk to introduce bacteria into the fresh skin wound resulting from tattooing and may be a risk factor for post-tattoo infections.

Biocide Susceptibility and Antimicrobial Resistance of Escherichia coli Isolated from Swine Feces, Pork Meat and Humans in Germany.

Phenotypic susceptibility testing of Escherichia (E.) coli is an essential tool to gain a better understanding of the potential impact of biocide selection pressure on antimicrobial resistance. We, therefore, determined the biocide and antimicrobial susceptibility of 216 extended-spectrum ß-lactamase-producing (ESBL) and 177 non-ESBL E. coli isolated from swine feces, pork meat, voluntary donors and inpatients and evaluated associations between their susceptibilities. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of benzalkonium chloride, chlorhexidine digluconate (CHG), chlorocresol (PCMC), glutaraldehyde (GDA), isopropanol (IPA), octenidine dihydrochloride and sodium hypochlorite (NaOCl) showed unimodal distributions, indicating the absence of bacterial adaptation to biocides due to the acquisition of resistance mechanisms. Although MIC95 and MBC95 did not vary more than one doubling dilution step between isolates of porcine and human origin, significant differences in MIC and/or MBC distributions were identified for GDA, CHG, IPA, PCMC and NaOCl. Comparing non-ESBL and ESBL E. coli, significantly different MIC and/or MBC distributions were found for PCMC, CHG and GDA. Antimicrobial susceptibility testing revealed the highest frequency of resistant E. coli in the subpopulation isolated from inpatients. We observed significant but weakly positive correlations between biocide MICs and/or MBCs and antimicrobial MICs. In summary, our data indicate a rather moderate effect of biocide use on the susceptibility of E. coli to biocides and antimicrobials.

Towards a Harmonized Terminology: A Glossary for Biocide Susceptibility Testing.

Disinfection is a key strategy to reduce the burden of infections. The contact of bacteria to biocides-the active substances of disinfectants-has been linked to bacterial adaptation and the development of antimicrobial resistance. Currently, there is no scientific consensus on whether the excessive use of biocides contributes to the emergence and spread of multidrug resistant bacteria. The comprehensive analysis of available data remains a challenge because neither uniform test procedures nor standardized interpretive criteria nor harmonized terms are available to describe altered bacterial susceptibility to biocides. In our review, we investigated the variety of criteria and the diversity of terms applied to interpret findings in original studies performing biocide susceptibility testing (BST) of field isolates. An additional analysis of reviews summarizing the knowledge of individual studies on altered biocide susceptibility provided insights into currently available broader concepts for data interpretation. Both approaches pointed out the urgent need for standardization. We, therefore, propose that the well-established and approved concepts for interpretation of antimicrobial susceptibility testing data should serve as a role model to evaluate biocide resistance mechanisms on a single cell level. Furthermore, we emphasize the adaptations necessary to acknowledge the specific needs for the evaluation of BST data. Our approach might help to increase scientific awareness and acceptance.

Unsuspected Positive Cultures in Planned Aseptic Revision Knee or Hip Arthroplasty-Risk Factors and Impact on Survivorship.

BACKGROUND: Unsuspected positive cultures (UPCs) may be found at the time of planned aseptic revision surgery and have previously been associated with decreased implant survival. However, reported rates vary greatly, and potential risk factors are widely unknown. This study investigates the rate of UPCs, implant survival, as well as potential risk factors for UPCs and revision. METHODS: This is a retrospective, single-center study on 434 cases of total hip arthroplasty (THA 277) and knee revision arthroplasty (TKA 157) performed between 2010 and 2017. Microbiology culture results, revision-free survival (RFS) and infection-free (IFS) survival, and the potential impact of UPCs as well as patient- or procedure-related risk factors were investigated at a minimum follow-up of 24 months (median 41 months). RESULTS: A total of 37% of patients had a positive culture (160/434). 27% (119/434) had a solitary positive culture, and 9% (41/434) had ≥2 positive cultures of the same species. Most commonly, coagulase-negative staphylococci were cultured. Overall, 32% (13/41) of patients with ≥2 positive cultures underwent revision for infection compared with 6% (17/274) of patients with negative cultures. RFS and IFS were reduced if ≥2 cultures were positive. Single positive cultures had no impact on RFS or IFS. Male sex, elevated serum C-reactive protein, and obesity were independently associated with finding ≥2 UPCs in revision THA. CONCLUSION: Appearance of ≥2 UPCs in aseptic revision leads to reduced RFS and IFS. Males, obese patients, and patients with elevated serum C-reactive protein planned for THA revision should be considered for extended diagnostics to rule out periprosthetic joint infection.

Total joint replacement of the hip and knee in patients with arthrogryposis multiplex congenita: a report of six joints.

INTRODUCTION: Arthrogryposis multiplex congenita (AMC) is a rare congenital condition that leads to severe joint contractures and deformities. As painful joint dysplasia and degeneration might develop over time, total joint replacement (TJR) can be a potential treatment option for these patients. The aim of this study is to investigate functional results, implant survivorship and potential complications in patients with AMC who undergo hip or knee arthroplasty. MATERIALS AND METHODS: We retrospectively identified six TJR in three patients at a single centre performed between 2006 and 2019. The median patient age at surgery was 23 years and the median follow-up period was 69 (IQR 55-99) months. We analysed surgical technique, implant survivorship and complications as well as functional outcome determined by pain reported on the Numerical Rating Scale (NRS), patient-reported outcome scores [Oxford Hip Score (OHS), Harris Hip score (HHS), Oxford Knee Score (OKS)], range of motion and ambulatory status. Depending on data distribution means with ranges and median with interquartile range were compared with the Wilcoxon signed rank test or Student's t test. The level of significance was defined at < 0.05. RESULTS: In hips, the mean range of motion in flexion/extension (52° vs. 85°, p = 0.014) and in rotation (28° vs. 68°, p = 0.02) as well as mean pain score on the NRS (8.5 vs. 0, p = 0.001), OHS (9 vs. 26, p = 0.031) and HHS (17 vs. 52, p = 0.007) significantly improved. In knees, mean range of motion (55° vs. 93°, p = 0.403), mean pain score on the NRS (0 vs. 7) and the OKS (2 vs. 21) also improved. While the ambulatory status did not change, the patients who were wheelchair dependent reported less problems with transfers to a bed or chair and the patient who ambulated reported an improved walking distance. One total knee arthroplasty (TKA) underwent revision for an acute, late infection 155 months following the initial surgery. CONCLUSIONS: TJR is a safe procedure in patients with AMC that effectively improves function and reduces pain irrespective of preoperative ambulatory status.

Revision TKA with a distal femoral replacement is at high risk of reinfection after two-stage exchange for periprosthetic knee joint infection.

PURPOSE: Megaprosthetic distal femoral reconstruction (DFR) is a limb-salvage procedure to address bone loss following two-stage revision for periprosthetic knee joint infection (PJI). The purpose of this study was to analyze the survival of DFR compared to hinged total knee arthroplasty (TKA). It was hypothesized that DFR was associated with a poorer survival. METHODS: In this retrospective single-center study, 97 subjects who underwent two-stage revision of chronic knee PJI were included. Among these, 41 were DFR. The diagnosis of PJI was established using the Musculoskeletal Infection Society (MSIS) criteria. Implant survival was calculated using Kaplan-Meier method and compared with the log-rank test as well as multivariate Cox regression at a minimum follow-up period of 24 months. RESULTS: The median follow-up period was 59 (interquartile range (IQR) 45-78) months. Overall, 24% (23/97) of patients required revision surgery for infection. The infection-free survival of rotating hinge revision TKA was 93% (95% Confidence Interval (CI) 86-100%) at five years compared to 50% (95% CI 34-66%) for DFR. In multivariate analysis, the risk factors for reinfection were DFR reconstruction (HR 4.7 (95% CI 1-22), p = 0.048), length of megaprosthesis (HR 1.006 (95% CI 1.001-1.012), p = 0.032) and higher BMI (HR 1.066, 95% CI 1.018-1.116), p = 0.007). 10% (4/41) of patients undergoing DFR underwent amputation to treat recurrent infection. CONCLUSION: Megaprosthetic DFR as part of a two-stage exchange for PJI is a salvage treatment that has a high risk for reinfection compared to non-megaprosthetic TKA. Patients must therefore be counseled accordingly. LEVEL OF EVIDENCE: Retrospective observational study, Level IV.

[Typing and particle analysis of squeaking hip endoprostheses : First histopathological analysis to examine the squeaking pathogenesis of ceramic-on-ceramic bearings]. / Typisierung und Partikelanalyse von Squeaking-Hüftendoprothesen : Erste histopathologische Analyse zur Untersuchung der Squeaking-Pathogenese von Keramik-Keramik-Gleitpaarungen.

BACKGROUND: Since the use of ceramic-on-ceramic (CoC) hip endoprostheses complications in the form of squeaking noises have occasionally occurred. OBJECTIVES: This is the first histopathological analysis of the synovia-like interface membrane (SLIM) of ceramic squeaking hip endoprostheses with the aim to gain new insights into the squeaking pathogenesis. MATERIALS AND METHODS: Seven CoC hip endoprostheses with squeaking pathogenesis are analyzed by SLIM consensus classification, particle algorithm, CD3 quantification, semiquantitative CD68 macrophages, Oil-Red positive macrophages, hemosiderin evaluation and in two cases by energy dispersive X­ray spectroscopy (EDX). RESULTS: In 1733 hip joint prosthesis pathology cases, a squeaking revision incidence of 0.40% was determined. In addition to SLIM type I (1/7), only SLIM type IV (6/7) was detected. 4/7 CoC cases showed combinations of micro, macro and, for the first time, supramacro (166.5 µm) ceramic wear particles. The EDX analysis confirmed the ceramic and an additional metallic abrasion. Increased focal concentrated low inflammatory markers (CD3/CD68) with hemosiderin (5/7) and lipid depositions (Oil-Red positive macrophages) (6/7) occurred. CONCLUSIONS: A pathogenetic connection between SLIM type I/IV and squeaking can be assumed. SLIM types showed a partly light microscopic ceramic particle-dependent, partly independent predominantly low-grade inflammation. Hemosiderin and Oil-Red positive macrophages are signs of synovial tissue damage and indicate biomechanical misload (impingement) and dysfunction as cause of the squeaking pathogenesis.

Does the Duration of Primary and First Revision Surgery Influence the Probability of First and Subsequent Implant Failures after Extremity Sarcoma Resection and Megaprosthetic Reconstruction?

Complications in megaprosthetic reconstruction following sarcoma resection are quite common. While several risk factors for failure have been explored, there is a scarcity of studies investigating the effect of the duration of surgery. We performed a retrospective study of 568 sarcoma patients that underwent megaprosthetic reconstruction between 1993 and 2015. Differences in the length of surgery and implant survival were assessed with the Kaplan-Meier method, the log-rank test and multivariate Cox regressions using an optimal cut-off value determined by receiver operating curves analysis using Youden's index. 230 patients developed a first and 112 patients a subsequent prosthetic failure. The median duration of initial surgery was 210 min. Patients who developed a first failure had a longer duration of the initial surgery (225 vs. 205 min, p = 0.0001). There were no differences in the probability of infection between patients with longer and shorter duration of initial surgery (12% vs. 13% at 5 years, p = 0.492); however, the probability of mechanical failure was higher in patients with longer initial surgery (38% vs. 23% at 5 years, p = 0.006). The median length of revision surgery for the first megaprosthetic failure was 101 min. Patients who underwent first revision for infection and did not develop a second failure had a longer median duration of the first revision surgery (150 min vs. 120 min, p = 0.016). A shorter length of the initial surgery appears beneficial, however, the notion that longer operating time increases the risk of deep infection could not be reproduced in our study. In revision surgery for infection, a longer operating time, possibly indicating a more thorough debridement, appears to be associated with a lower risk for subsequent revision.

The change in plasma D-dimer does not help to guide the timing of reimplantation in two stage exchange for periprosthetic joint infection.

D-dimer has been included in the criteria by the Musculoskeletal Infection Society in 2018 as a novel parameter to diagnose prosthetic joint infection (PJI). However, it is unclear how D-dimer levels change in between stages of a two-stage exchange. We prospectively investigated 30 patients who underwent a two-stage exchange using a spacer for PJI. D-Dimer, CRP and IL-6 were collected before first and second stage surgery and the difference (Δ) in between stages was calculated. The levels of plasma D-Dimer did not change from first to second stage surgery (2770 ng/ml (IQR, 1600-3770 ng/ml) versus 2340 ng/ml (IQR, 1270-4100 ng/ml); p = 0.8) while CRP (4.0 mg/dl (IQR, 1.7-5.5 mg/dl) versus 0.6 mg/dl (IQR, 0.5-0.8 mg/dl); p < 0.001) and IL-6 (21 pg/ml (IQR, 10-29 pg/ml) versus 6 pg/ml (4-9 pg/ml); p < 0.001) decreased. The ΔD-dimer between both stages was 300 ng/ml (range: - 2820 to 4280 ng/ml), the median ΔCRP was - 3.4 mg/dl (IQR, - 1.2 to - 4.8 mg/dl) and ΔIL-6 was - 13 pg/ml (IQR, - 4 to - 20 pg/ml). In 15 of 30 cases (50%) the D-dimer level increased between both stages, whereas the level of CRP (93%; 28/30) and IL-6 (96%; 28/29) decreased in most patients. As the level of serum D-dimers varies greatly, lacks a uniform decrease and does not identify persisting infection, surgeons should be cautious when using it at the timing of reimplantation.

Genetic but No Phenotypic Associations between Biocide Tolerance and Antibiotic Resistance in Escherichia coli from German Broiler Fattening Farms.

Biocides are frequently applied as disinfectants in animal husbandry to prevent the transmission of drug-resistant bacteria and to control zoonotic diseases. Concerns have been raised, that their use may contribute to the selection and persistence of antimicrobial-resistant bacteria. Especially, extended-spectrum ß-lactamase- and AmpC ß-lactamase-producing Escherichia coli have become a global health threat. In our study, 29 ESBL-/AmpC-producing and 64 NON-ESBL-/AmpC-producing E.coli isolates from three German broiler fattening farms collected in 2016 following regular cleaning and disinfection were phylogenetically characterized by whole genome sequencing, analyzed for phylogenetic distribution of virulence-associated genes, and screened for determinants of and associations between biocide tolerance and antibiotic resistance. Of the 30 known and two unknown sequence types detected, ST117 and ST297 were the most common genotypes. These STs are recognized worldwide as pandemic lineages causing disease in humans and poultry. Virulence determinants associated with extraintestinal pathogenic E.coli showed variable phylogenetic distribution patterns. Isolates with reduced biocide susceptibility were rarely found on the tested farms. Nine isolates displayed elevated MICs and/or MBCs of formaldehyde, chlorocresol, peroxyacetic acid, or benzalkonium chloride. Antibiotic resistance to ampicillin, trimethoprim, and sulfamethoxazole was most prevalent. The majority of ESBL-/AmpC-producing isolates carried blaCTX-M (55%) or blaCMY-2 (24%) genes. Phenotypic biocide tolerance and antibiotic resistance were not interlinked. However, biocide and metal resistance determinants were found on mobile genetic elements together with antibiotic resistance genes raising concerns that biocides used in the food industry may lead to selection pressure for strains carrying acquired resistance determinants to different antimicrobials.

MALDI-TOF MS and genomic analysis can make the difference in the clarification of canine brucellosis outbreaks.

Brucellosis is one of the most common bacterial zoonoses worldwide affecting not only livestock and wildlife but also pets. Canine brucellosis is characterized by reproductive failure in dogs. Human Brucella canis infections are rarely reported but probably underestimated due to insufficient diagnostic surveillance. To improve diagnostics, we investigated dogs in a breeding kennel that showed clinical manifestations of brucellosis and revealed positive blood cultures. As an alternative to the time-consuming and hazardous classical identification procedures, a newly developed species-specific intact-cell matrix-assisted laser desorption/ionization-time of flight mass spectrometry analysis was applied, which allowed for rapid identification of B. canis and differentiation from closely related B. suis biovar 1. High-throughput sequencing and comparative genomics using single nucleotide polymorphism analysis clustered our isolates together with canine and human strains from various Central and South American countries in a distinct sub-lineage. Hence, molecular epidemiology clearly defined the outbreak cluster and demonstrated the endemic situation in South America. Our study illustrates that MALDI-TOF MS analysis using a validated in-house reference database facilitates rapid B. canis identification at species level. Additional whole genome sequencing provides more detailed outbreak information and leads to a deeper understanding of the epidemiology of canine brucellosis.

A Proof of Principle for the Detection of Viable Brucella spp. in Raw Milk by qPCR Targeting Bacteriophages.

Brucellosis is still a global health issue, and surveillance and control of this zoonotic disease in livestock remains a challenge. Human outbreaks are mainly linked to the consumption of unpasteurized dairy products. The detection of human pathogenic Brucella species in food of animal origin is time-consuming and laborious. Bacteriophages are broadly applied to the typing of Brucella isolates from pure culture. Since phages intracellularly replicate to very high numbers, they can also be used as specific indicator organisms of their host bacteria. We developed a novel real-time PCR (qPCR) assay targeting the highly conserved helicase sequence harbored in all currently known Brucella-specific lytic phages. Quality and performance tests determined a limit of detection of <1 genomic copy/µL. In raw milk artificially contaminated with Brucella microti, Izv phages were reliably detected after 39 h of incubation, indicating the presence of viable bacteria. The qPCR assay showed high stability in the milk matrix and significantly shortened the time to diagnosis when compared to traditional culture-based techniques. Hence, our molecular assay is a reliable and sensitive method to analyze phage titers, may help to reduce the hands-on time needed for the screening of potentially contaminated food, and reveals infection risks without bacterial isolation.

Clinical use of linezolid in periprosthetic joint infections - a systematic review.

Introduction: The most common causative organism in periprosthetic joint infections (PJIs) is Gram-positive bacteria that are increasingly drug resistant. In these cases the use of linezolid may be warranted. However, there are conflicting reports regarding its role in antibiotic treatment of PJIs. The aim of this review is to gather and analyze clinical results and treatment details on linezolid in patients with PJIs. Methods: In August 2019, a comprehensive literature search using MEDLINE (Pubmed and Ovid) and Cochrane Library was performed. A total of 504 records were screened, and a total of 16 studies including 372 patients treated with linezolid for a PJI were included in this review based on the PRISMA criteria and after quality analysis using the MINOR score and Newcastle-Ottawa scale, as well as assessing level of evidence. Pooling analysis as well as descriptive analysis was performed. Results: Based on the results from the studies included, infection control was achieved in 80 % (range 30 %-100 %) of patients after a mean follow-up period of 25 (range 2-66) months. The mean duration of treatment was 58 d intravenous and orally at a median dose of 600 mg bis in die (b.i.d.) (range 400-900 b.i.d.). A combination therapy with rifampicin was used in 53 % of patients. MRSA (methicillin-resistant Staphylococcus aureus) infections were present in 29 % and resistant CoNS (coagulase-negative Staphylococcus) in 46 %. Adverse effects occurred in 33 % of cases, mostly anemia, thrombocytopenia and gastrointestinal complaints leading to treatment discontinuation in 9 %. However, great heterogeneity was found with respect to surgical treatment, diagnosis of infection and indication for linezolid. Discussion: Linezolid is an appropriate option for treatment of resistant Gram-positive organisms in PJIs. Most commonly 600 mg b.i.d. is used, and a combination with rifampicin appears feasible although one must consider individual increases in doses in these cases. However, adverse effects are common and there are limited data for long-term use and optimal antibiotic combinations or individual doses.

Comparing the Diagnostic Value of Serum D-Dimer to CRP and IL-6 in the Diagnosis of Chronic Prosthetic Joint Infection.

Introduction: D-dimer is a diagnostic criterion for periprosthetic joint infection (PJI) of the Musculoskeletal Infection Society (MSIS) in 2018. The aim of this study was to evaluate the serum D-dimer values in comparison to C-reactive protein (CRP) and interleukin-6 (IL-6) for the diagnosis of PJI. Materials and Methods: We included 119 patients (50 women, 69 men; 71 knees, 48 hips) undergoing revision arthroplasty with preoperative assessment of CRP, IL-6, and serum D-dimer. Cases were classified as infected or aseptic based on the MSIS criteria of 2018. Receiver operating curves and Youden's index were used to define an ideal cut-off value and sensitivity and specificity for the individual parameters, and respective combinations were calculated using cross-tables. Results: The median D-dimer level (2320 vs. 1105 ng/mL; p < 0.001), the median CRP level (4.0 vs. 0.5 mg/dL; p < 0.001), and the median IL-6 level (21.0 vs. 5.0 pg/mL; p < 0.001) were significantly higher in the group of PJI compared to the group with aseptic failure. The calculated optimal cut-off values were 2750 ng/mL (AUC 0.767) for D-dimer, 1.2 mg/dL (AUC 0.914) for CRP, and 10.0 pg/mL (AUC 0.849) for IL-6. D-dimer showed a sensitivity of 38% and specificity of 94%, whereas the CRP and IL-6 had sensitivities of 88% and 76%, and specificities of 87% and 92%, respectively. Conclusion: In comparison with CRP and IL-6, serum D-dimer showed low sensitivity and specificity in our cohort. While CRP and IL-6 combination had the highest sensitivity, a combination of Il-6 and D-dimer or CRP and IL-6 had the highest specificity.

Backtracking and forward checking of human listeriosis clusters identified a multiclonal outbreak linked to Listeria monocytogenes in meat products of a single producer.

Due to its high case fatality rate, foodborne listeriosis is considered a major public health concern worldwide. We describe one of the largest listeriosis outbreaks in Germany with 83 cases of invasive listeriosis between 2013 and 2018. As part of the outbreak investigation, we identified a highly diverse Listeria monocytogenes population at a single producer of ready-to-eat meat products. Strikingly, the extensive sampling after identification of a first match between a cluster of clinical isolates and a food isolate allowed for a linkage between this producer and a second, previously unmatched cluster of clinical isolates. Bacterial persistence in the processing plant and indications of cross-contamination events explained long-term contamination of food that led to the protracted outbreak. Based on screening for virulence factors, a pathogenic phenotype could not be ruled out for other strains circulating in the plant, suggesting that the outbreak could have been even larger. As most isolates were sensitive to common biocides used in the plant, hard to clean niches in the production line may have played a major role in the consolidation of the contamination. Our study demonstrates how important it is to search for the origin of infection when cases of illness have occurred (backtracking), but also clearly highlights that it is equally important to check whether a contamination at food or production level has caused disease (forward checking). Only through this two-sided control strategy, foodborne disease outbreaks such as listeriosis can be minimized, which could be a real improvement for public health.

Microbially competent 3D skin: a test system that reveals insight into host-microbe interactions and their potential toxicological impact.

The skin`s microbiome is predominantly commensalic, harbouring a metabolic potential far exceeding that of its host. While there is clear evidence that bacteria-dependent metabolism of pollutants modulates the toxicity for the host there is still a lack of models for investigating causality of microbiome-associated pathophysiology or toxicity. We now report on a biologically characterised microbial-skin tissue co-culture that allows studying microbe-host interactions for extended periods of time in situ. The system is based on a commercially available 3D skin model. In a proof-of-concept, this model was colonised with single and mixed cultures of two selected skin commensals. Two different methods were used to quantify the bacteria on the surface of the skin models. While Micrococcus luteus established a stable microbial-skin tissue co-culture, Pseudomonas oleovorans maintained slow continuous growth over the 8-day cultivation period. A detailed skin transcriptome analysis showed bacterial colonisation leading to up to 3318 significant changes. Additionally, FACS, ELISA and Western blot analyses were carried out to analyse secretion of cytokines and growth factors. Changes found in colonised skin varied depending on the bacterial species used and comprised immunomodulatory functions, such as secretion of IL-1α/ß, Il-6, antimicrobial peptides and increased gene transcription of IL-10 and TLR2. The colonisation also influenced the secretion of growth factors such as VFGFA and FGF2. Notably, many of these changes have already previously been associated with the presence of skin commensals. Concomitantly, the model gained first insights on the microbiome's influence on skin xenobiotic metabolism (i.e., CYP1A1, CYP1B1 and CYP2D6) and olfactory receptor expression. The system provides urgently needed experimental access for assessing the toxicological impact of microbiome-associated xenobiotic metabolism in situ.

Promising results of revision total hip arthroplasty using a hexagonal, modular, tapered stem in cases of aseptic loosening.

BACKGROUND: Modular stems are widely used in revision total hip arthroplasty (THA) with aseptic loosening being a common reason for revision. Despite the good results reported on the use of modular stem designs, there are only few studies focusing on aseptic revisions and few studies on a hexagonal stem design. The goal of this study is to determine stem survival, clinical and functional outcome along with possible risk factors for implant failure in aseptic revision THA. METHODS: We retrospectively identified 53 patients with aseptic THA revision using a modular hexagonal stem with a minimum follow-up of two years. Femoral bone loss, radiographic and clinical outcomes as well as function measured using the Harris Hip Score (HHS) was assessed. Patients' previous medical history was analyzed for comorbidities and the body mass index. Stem survival was calculated using the Kaplan-Meier method. Possible risk factors for implant failure were analyzed using the log-rank test. The median age at revision was 69 (IQR 62-73) with a median follow up of 74 months. RESULTS: Implant survival rates amounted to 90.4% at 3 and 5 years. The median HHS improved by 47 points (34 (IQR 22-47) vs 81 (IQR 59-90) p<0.001). There was a reduced implant survival after 5 years when the revision stem was used following a previous cemented stem (83.4% vs 100%, p = 0.04). CONCLUSION: A modular, hexagonal stem can be successfully used in aseptic revision THA with remarkable functional results and excellent survivorship. Revision of a cemented stem using this implant might result in reduced survival which must be considered when planning treatment.