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Coagulase-negative staphylococci (CoNS), members of the skin commensal microbiota, are increasingly associated with local or systemic infections due to a shift in patient populations in recent decades. Subsequently, more CoNS strains have been subjected to antibiotic susceptibility testing (AST), thus leading to the increased detection of teicoplanin resistance. However, data concerning teicoplanin resistance among CoNS strains remain limited, heterogeneous, and inconclusive. We collected 162 consecutive CoNS strains identified using Vitek-2 as teicoplanin-resistant and tested them with a range of AST methods. The results of standard and high inoculum broth microdilution (sBMD; hBMD), agar dilution (AD) after 24 h and 48 h incubation, standard and macrogradient diffusion strip (sGDT, MET), screening agar, and disc diffusion were compared to assess their robustness and to establish a diagnostic algorithm to detect teicoplanin resistance. sBMD was used as the reference method, and the lowest number of strains were teicoplanin-resistant using this method. sGDT and disc diffusion generated similar results to sBMD. Compared with sBMD, AD-24 h generated the lowest number of false teicoplanin-resistant strains, followed by hBMD, AD-48 h, and Vitek-2. sGDT, a fast, easy, affordable method in diagnostic settings, generated the highest rate of false teicoplanin-susceptible strains. Vitek-2 testing produced the highest number of teicoplanin-resistant strains. Only in two strains was the initial Vitek-2 teicoplanin resistance confirmed using five other AST methods. In conclusion, the different antibiotic susceptibility testing methods generated inconsistent, inconclusive, and discrepant results, thus making it difficult to establish a diagnostic algorithm for suspected teicoplanin resistance. Teicoplanin testing proved to be challenging and easily influenced by technical factors. This study aimed not only to raise awareness of teicoplanin resistance testing but also of the need for future studies focusing on the clinical efficacy of teicoplanin in relation to its susceptibility results.

A patient with oral squamous cell carcinoma (OSCC) underwent complex surgical tumor therapy, including the reconstruction of soft tissues using a radial forearm flap. Due to venous congestion that could only partly be resolved by revision surgery, leech therapy was started on the second postoperative day. The patient developed pneumonia and sepsis and died as a result of septic shock, despite having received targeted broad-spectrum antibiotic therapy since day 5. Aeromonas spp. were cultured from both the patient’s specimens and unused leeches. Biochemical identification and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) yielded inconsistent identification results. Finally, microbiological identification of Aeromonas spp. was performed via 16S rDNA sequencing and use of the basic local alignment search tool (BLAST), and strains from both the patient and the leeches were identified as Aeromonas veronii. Aeromonas spp. strains derived from the patient and leeches and independent laboratory strains were submitted to randomly amplified polymorphic DNA (RAPD) subtyping. RAPD of A. veronii strains from both sources revealed an identical pattern, strongly suggesting the transmission of A. veronii from the leeches to the patient. Physicians should be aware of the potential for severe lethal infections as a fatal side-effect of leech therapy in critically ill patients, which should be addressed using antibiotic prophylaxis.

Coagulase-negative staphylococci (CoNS), members of the skin commensal microbiota, are increasingly associated with local or systemic infections due to a shift in patient populations in recent decades. Subsequently, more CoNS strains have been subjected to antibiotic susceptibility testing (AST), thus leading to the increased detection of teicoplanin resistance. However, data concerning teicoplanin resistance among CoNS strains remain limited, heterogeneous, and inconclusive. We collected 162 consecutive CoNS strains identified using Vitek-2 as teicoplanin-resistant and tested them with a range of AST methods. The results of standard and high inoculum broth microdilution (sBMD; hBMD), agar dilution (AD) after 24 h and 48 h incubation, standard and macrogradient diffusion strip (sGDT, MET), screening agar, and disc diffusion were compared to assess their robustness and to establish a diagnostic algorithm to detect teicoplanin resistance. sBMD was used as the reference method, and the lowest number of strains were teicoplanin-resistant using this method. sGDT and disc diffusion generated similar results to sBMD. Compared with sBMD, AD-24 h generated the lowest number of false teicoplanin-resistant strains, followed by hBMD, AD-48 h, and Vitek-2. sGDT, a fast, easy, affordable method in diagnostic settings, generated the highest rate of false teicoplanin-susceptible strains. Vitek-2 testing produced the highest number of teicoplanin-resistant strains. Only in two strains was the initial Vitek-2 teicoplanin resistance confirmed using five other AST methods. In conclusion, the different antibiotic susceptibility testing methods generated inconsistent, inconclusive, and discrepant results, thus making it difficult to establish a diagnostic algorithm for suspected teicoplanin resistance. Teicoplanin testing proved to be challenging and easily influenced by technical factors. This study aimed not only to raise awareness of teicoplanin resistance testing but also of the need for future studies focusing on the clinical efficacy of teicoplanin in relation to its susceptibility results. [ABSTRACT FROM AUTHOR]

In accordance with worldwide data, the Robert Koch Institute (RKI) has reported a constant increase of syphilis cases in Germany over the past decade. We analysed the data of all patients, referred to a Department of Ophthalmology in a tertiary referral centre in Düsseldorf, Germany between 2008 and 2019, who were tested for syphilis. The epidemiologic, demographic, clinical, diagnostic and therapeutic data were retrieved from the records and evaluated in a retrospective, descriptive, non-comparative study. Syphilis serology was positive in 32/1840 (1.7%) patients, and was evenly distributed over this period. 26 (81.3%) were male, 19 (59.4%) belonged to a risk group. Ocular syphilis was the primary diagnosis for 29 patients (90.6%). The most frequent manifestation was uveitis (n = 20, 62.5%). By the end of therapy, 19 patients (59.4%) had an improved visual acuity. The incidence of ocular syphilis cases has remained stable over the last decade. [ABSTRACT FROM AUTHOR]

A patient with oral squamous cell carcinoma (OSCC) underwent complex surgical tumor therapy, including the reconstruction of soft tissues using a radial forearm flap. Due to venous congestion that could only partly be resolved by revision surgery, leech therapy was started on the second postoperative day. The patient developed pneumonia and sepsis and died as a result of septic shock, despite having received targeted broad-spectrum antibiotic therapy since day 5. Aeromonas spp. were cultured from both the patient's specimens and unused leeches. Biochemical identification and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) yielded inconsistent identification results. Finally, microbiological identification of Aeromonas spp. was performed via 16S rDNA sequencing and use of the basic local alignment search tool (BLAST), and strains from both the patient and the leeches were identified as Aeromonas veronii. Aeromonas spp. strains derived from the patient and leeches and independent laboratory strains were submitted to randomly amplified polymorphic DNA (RAPD) subtyping. RAPD of A. veronii strains from both sources revealed an identical pattern, strongly suggesting the transmission of A. veronii from the leeches to the patient. Physicians should be aware of the potential for severe lethal infections as a fatal side-effect of leech therapy in critically ill patients, which should be addressed using antibiotic prophylaxis. [ABSTRACT FROM AUTHOR]

Aim: To identify dominant microorganisms in root filled teeth with apical periodontitis by Pan‐PCRs in comparison with a culture‐dependent approach, focusing on fungal species profiling. Methodology: The root filling material (gutta‐percha) removed from 42 teeth with periapical radiolucencies undergoing root canal retreatments was analysed by molecular genetics techniques. Real‐Time Pan‐PCRs were conducted for the diagnosis of predominant bacteria (targeting 16S rDNA) and fungi (targeting ITS1‐2 region). Identification of microorganisms was performed by Sanger sequencing of the PCR products and BLAST analysis. Additionally, subgingival plaque samples were collected and cultured to review the composition of the microbial flora. The McNemar test and the repeated measures anova were used for statistical analyses (significance level was set at P < 0.05). Results: Overall, 42/42 plaque samples had bacterial growth, whereas 32/42 gutta‐percha samples had bacterial growth with a dominance of Streptococcus spp. (12/42) and Enterococcus faecalis (9/42). The mean number of bacterial taxa per gutta‐percha sample was 1.6 cultivatable taxa, significantly lower than in the plaque sample that had six taxa/sample (P < 0.001). Fungus‐specific cultures were negative for gutta‐percha samples, and only one plaque sample had growth of a fungus. In total, 36/42 plaque samples were positive in bacterial Pan‐PCRs. In bacterial Pan‐PCRs of 31/42 gutta‐percha samples, dominant microorganisms were identified including Streptococcus spp. (5/42) and E. faecalis (4/42). Moreover, in 7/42 gutta‐percha samples, DNA of bacteria which are difficult‐to‐cultivate in microbiology routine culture (Acinetobacter,Pyramidobacter,Bacteroidetes,Synergistes,Atopobium and Pseudoramibacter) was found. DNA of Candida spp. was detected in 5/42 root canals by fungal Pan‐PCR (1/5) and genus‐specific Candida‐PCR (5/5). Conclusions: Pan‐PCR assays remain appropriate as a broad‐range approach for the detection of a dominant pathogen in gutta‐percha samples which have less diverse microbial composition. The molecular genetic Pan‐PCR approach has the advantage of detecting microorganisms that are as‐yet‐uncultivable or difficult‐to‐cultivate and should be therefore complement conventional microbiological diagnostics. [ABSTRACT FROM AUTHOR]

In vivo confocal microscopy (IVCM) is a non-invasive imaging technique that allows morphological analysis as a diagnostic approach of the cornea in real time, thus providing a suspected diagnosis of fungal or amoebic keratitis immediately, whereas culture or PCR require several days or even weeks. Since these infections are rare, it is difficult for ophthalmologists to gain the experience necessary to differentiate infection from normal findings or artefacts. The purpose of this project was to establish a simulator, on which physicians could practice as well as acquiring a database of IVCM images of fungal or amoebic keratitis and respective analyses. An IVCM simulator was set up with cadaver human corneas, infected with either acanthamoeba, candida or aspergillus. Twenty-one ophthalmologists were trained in IVC microscopy first in a Dry Lab, then practically on the simulator. For evaluation, the participants were asked to fill out a standardized questionnaire, with a pre- and post-course self-assessment. The self-assessed theoretical and practical skills in differentiating infectious from non-infectious keratitis in IVCM significantly increased (p = 0.0001, p = 0.0002, respectively). The barrier to use this technique decreased (p = 0.0474). A very simple protocol based on a model of ex vivo corneal mycotic and amoebic infections can be used to train novices in the structured approach and diagnostic use of IVCM for corneal infections. [ABSTRACT FROM AUTHOR]