Timely approaches to identify probiotic species of the genus Lactobacillus

Over the past decades the use of probiotics in food has increased largely due to the manufacturer's interest in placing "healthy" food on the market based on the consumer's ambitions to live healthy. Due to this trend, health benefits of products containing probiotic strains such as lactobacilli are promoted and probiotic strains have been established in many different products with their numbers increasing steadily. Probiotics are used as starter cultures in dairy products such as cheese or yoghurts and in addition they are also utilized in non-dairy products such as fermented vegetables, fermented meat and pharmaceuticals, thereby, covering a large variety of products.To assure quality management, several pheno-, physico- and genotyping methods have been established to unambiguously identify probiotic lactobacilli. These methods are often specific enough to identify the probiotic strains at genus and species levels. However, the probiotic ability is often strain dependent and it is impossible to distinguish strains by basic microbiological methods.Therefore, this review aims to critically summarize and evaluate conventional identification methods for the genus Lactobacillus, complemented by techniques that are currently being developed.1\. Auflag

ESBL-plasmids carrying toxin-antitoxin systems can be “cured” of wild-type Escherichia coli using a heat technique

Plasmid-encoded extended-spectrum beta-lactamase (ESBL)-enzymes are frequently produced by Escherichia coli. Several ESBL-plasmids contain genes for toxin- antitoxin (TA) systems, which assure the maintenance of plasmids in bacteria and prevent the cells from "post-segregational killing". These systems limit options to "cure" plasmids of ESBL-wild-type strains due to the death of the bacterial cells. A helpful tool to understand the role of ESBL-plasmids in the dissemination of pandemic multi-resistant E. coli are ESBL- plasmid-"cured"-variants (PCVs) and their comparison to ESBL-wild-type strains. The purpose of this study was to construct PCVs of ESBL-wild-type E. coli strains despite the presence of genes for TA systems. Using enhanced temperatures and brain-heart-infusion broth it was possible to construct viable PCVs of wild-type ESBL-E. coli strains. The occurrence of TA system- genes including hok/sok, srnB/C, vagC/D, pemI/K on ESBL-plasmids of replicon types FIA or FIB was demonstrated by bioinformatic analyses. The loss of the plasmid and the genetic identity of PCV and corresponding wild-type strain was confirmed via different methods including plasmid-profile-analysis, pulsed- field gel electrophoresis and bioinformatics using generated whole genome data of the strains. This short report describes the successful construction of viable PCVs of ESBL-wild-type E. coli strains. The results are hence surprising due to the fact that all "cured" ESBL-plasmids contained at least one complete toxin-antitoxin system, whose loss would normally mean the death of bacterial cells

Perceived Access to Health Care Services and Relevance of Telemedicine during the COVID-19 Pandemic in Germany

During the COVID-19 pandemic in Germany, non-pharmaceutical interventions were imposed to contain the spread of the virus. Based on cross-sectional waves in March, July and December 2020 of the COVID-19 Snapshot Monitoring (COSMO), the present study investigated the impact of the introduced measures on the perceived access to health care. Additionally, for the wave in December, treatment occasion as well as utilization and satisfaction regarding telemedicine were analysed. For 18–74-year-old participants requiring medical care, descriptive and logistic regression analyses were performed. During the less strict second lockdown in December, participants reported more frequently ensured access to health care (91.2%) compared to the first lockdown in March (86.8%), but less frequently compared to July (94.2%) during a period with only mild restrictions. In December, main treatment occasions of required medical appointments were check-up visits at the general practitioner (55.2%) and dentist (36.2%), followed by acute treatments at the general practitioner (25.6%) and dentist (19.0%), treatments at the physio-, ergo- or speech therapist (13.1%), psychotherapist (11.9%), and scheduled hospital admissions or surgeries (10.0%). Of the participants, 20.0% indicated utilization of telemedical (15.4% telephone, 7.6% video) consultations. Of them, 43.7% were satisfied with the service. In conclusion, for the majority of participants, access to medical care was ensured during the COVID-19 pandemic; however, access slightly decreased during phases of lockdown. Telemedicine complemented the access to medical appointments.Peer Reviewe

Phylogenetic and Molecular Analysis of Food-Borne Shiga Toxin-Producing Escherichia coli

Seventy-five food-associated Shiga toxin-producing Escherichia coli (STEC) strains were analyzed by molecular and phylogenetic methods to describe their pathogenic potential. The presence of the locus of proteolysis activity (LPA), the chromosomal pathogenicity island (PAI) PAI ICL3, and the autotransporter- encoding gene sabA was examined by PCR. Furthermore, the occupation of the chromosomal integration sites of the locus of enterocyte effacement (LEE), selC, pheU, and pheV, as well as the Stx phage integration sites yehV, yecE, wrbA, z2577, and ssrA, was analyzed. Moreover, the antibiotic resistance phenotypes of all STEC strains were determined. Multilocus sequence typing (MLST) was performed, and sequence types (STs) and sequence type complexes (STCs) were compared with those of 42 hemolytic-uremic syndrome (HUS)-associated enterohemorrhagic E. coli (HUSEC) strains. Besides 59 STs and 4 STCs, three larger clusters were defined in this strain collection. Clusters A and C consist mostly of highly pathogenic eae-positive HUSEC strains and some related food-borne STEC strains. A member of a new O26 HUS-associated clone and the 2011 outbreak strain E. coli O104:H4 were found in cluster A. Cluster B comprises only eae-negative food-borne STEC strains as well as mainly eae-negative HUSEC strains. Although food-borne strains of cluster B were not clearly associated with disease, serotypes of important pathogens, such as O91:H21 and O113:H21, were in this cluster and closely related to the food-borne strains. Clonal analysis demonstrated eight closely related genetic groups of food-borne STEC and HUSEC strains that shared the same ST and were similar in their virulence gene composition. These groups should be considered with respect to their potential for human infection