In order to further accelerate pathogen identification from positive blood cultures (BC), various sample preparation protocols to identify bacteria with MALDI-TOF MS directly from positive BCs have been developed. We evaluated an in-house method in comparison to the Sepsityper^® Kit (Bruker Daltonics, Bremen, Germany) as well as the benefit of an on-plate formic acid extraction step following positive signal by the BACTEC^TM FX system. Confirmation of identification was achieved using subcultured growing biomass used for MALDI-TOF MS analysis. A total of 113 monomicrobial positive BCs were analyzed. The rates of Gram-positive bacteria correctly identified to the genus level using in-house method and Sepsityper^® Kit were 63.3% (38/60) and 81.7% (49/60), respectively (p = 0.025). Identification rates at species level for Gram-positive bacteria with in-house method and Sepsityper^® kit were 30.0% (18/60) and 66.7% (40/60), respectively (p < 0.001). Identification rates of Gram-negative bacteria were similar with the in-house method and Sepsityper^® Kit. Additional on-plate formic acid extraction demonstrated significant improvement in the identification rate of Gram-positive bacteria at both genus and species level for both in-house (p = 0.001, p < 0.001) and Sepsityper^® Kit methods (p = 0.007, p < 0.001). Our in-house method is a candidate for laboratory routines with Sepsityper^® Kit as a back-up solution when identification of Gram-positive bacteria is unsuccessful.

While smoking is recognized as one of the factors for the development and progression of periodontal diseases, a relation between the composition of the subgingival microbiota and smoking is yet to be elucidated. The aim of this study was to investigate the prevalence of subgingival bacteria in young smokers and non-smokers without clinical signs of periodontal disease. In this cross-sectional study, performed at the Department of Pharmacology, School of Dental Medicine, University of Zagreb, we enrolled 32 periodontally healthy smokers and 32 non-smokers, aged 25–35 years old. The number of oral bacteria and the prevalence of particular bacteria were assessed for each subject. Subgingival plaque samples were collected with sterile paper points from two first molars for microbiological analyses with MALDI-TOF mass spectrometry. In smokers, a significantly higher prevalence of Actinomyces odontolyticus was observed compared to non-smokers, and a significantly lower prevalence of Streptococcus sanguinis was observed compared to non-smokers. Smoking affects the composition of subgingival microbiota, either via depletion of beneficial bacteria or the increase in pathogenic bacteria.

Two Enterobacter hormaechei isolates harbouring three carbapenemase genes each, were isolated from two patients from different ICUs at University Hospital Centre Zagreb, Croatia, which is to our knowledge, the first report of triple carbapenemase (bla(VIM-2), bla(NDM-1), and bla(OXA-48)) co-existence in E. hormachei strains and also among Enterobacterales members in Croatia. Antimicrobial susceptibility testing showed susceptibility only to colistin and amikacin. The production of carbapenemases was phenotypically tested by immunochromatographic assay and confirmed by PCR. Detailed analysis by Whole Genome Sequencing (WGS) of short reads by Illumina and long reads by Oxford Nanopore Technologies (ONT) was additionally performed and showed that both isolates belonged to ST200. They were separated by 98 Single Nucleotide Polymorphisms (SNPs) having variations in the number of bla(VIM-2) genes on the chromosome, the number of bla(NDM-1) genes on the plasmid, non-identical bla(NDM-1) plasmids, different plasmid content in general, and only one isolate carried a 94 kb prophage.

The aim of the present study was to investigate whether real-time polymerase chain reaction (PCR) has a low identification rate for samples with low acid-fast bacilli (AFB) grades, including those obtained using bronchoscopy. When 50 smear-positive samples were compared by AFB score and PCR result, PCR was 100% successful in identifying AFB 2+ and AFB 3+ samples. However, only 14 of 26 (52%) AFB 1+ samples were identified. In paucibacillary smear-positive samples, PCR is not reliable enough to exclude tuberculosis, but in smear-positive patients with high AFB grades PCR can immediately change the clinical management of the disease.

Background/Objectives: The aetiology of Bell’s palsy remains unclear and is typically diagnosed by exclusion. This study investigated the potential role of neurotropic viruses and explored the relationship between facial nerve impairment and vestibular dysfunction to improve the understanding of the condition. Methods: Antibodies against herpes simplex virus (HSV) and varicella-zoster virus (VZV) were assessed using ELISA. Vestibular function was evaluated through computerised videonystagmography, rotatory chair, and clinical vestibulospinal assessments. Facial nerve lesion localisation was determined by stapedial reflex testing. Fisher’s exact test was used for statistical analysis. Results: Of 51 patients with Bell’s palsy, 62.7% exhibited vestibular dysfunction, and 70.6% were IgG-positive for at least one neurotropic virus. Vestibular impairment was significantly more common in seropositive patients. Statistically significant associations were observed between vestibular dysfunction and viral IgG seropositivity (p < 0.0001), the severity of vestibular dysfunction and facial paresis (p = 0.0126), and the side of vestibular impairment and the side of facial palsy (p < 0.0001), with 90.6% of cases showing ipsilateral involvement. Conclusions: These findings support the hypothesis that neurotropic viruses may act as a common pathological factor in both Bell’s palsy and associated vestibular dysfunction.