In 1970, the seventh pandemic of cholera (7 P) reached both Africa and Europe. Between 1970 and 2011, several European countries reported cholera outbreaks of a few to more than 2,000 cases. We report here a whole-genome analysis of 1,324 7 P V. cholerae El Tor (7 PET) isolates, including 172 from autochthonous sporadic or outbreak cholera cases occurring between 1970 and 2011 in Europe, providing insight into the spatial and temporal spread of this pathogen across Europe. In this work, we show that the 7 PET lineage was introduced at least eight times into two main regions: Eastern and Southern Europe. Greater recurrence of the disease was observed in Eastern Europe, where it persisted until 2011. It was introduced into this region from Southern Asia, often circulating regionally in the countries bordering the Black Sea, and in the Middle East before reaching Eastern Africa on several occasions. In Southern Europe, the disease was mostly seen in individual countries during the 1970s and was imported from North and West Africa, except in 1994, when cholera was imported into Albania and Italy from the Black Sea region. These results shed light on the geographic course of cholera during the seventh pandemic and highlight the role of humans in its global dissemination.

IntroductionAt the beginning of 2016, an increase in paediatric haemolytic uremic syndrome (HUS) cases was observed in Romania. The microbiological investigations allowed isolation of Shiga toxin-producing Escherichia coli (STEC) O26 as the causative agent from most cases. Methods: An enhanced national surveillance of HUS and severe diarrhoea was established across the country following the identification of the first cases and was carried out until August 2016. A total of 15 strains were isolated from 10 HUS and five diarrhoea cases. Strains were characterised by virulence markers (i.e. stx type/subtype, eae, ehxA genes), phylogroup, genetic relatedness and clonality using PCR-based assays, PFGE and multilocus sequence typing (MLST). The first six strains were further characterised by whole genome sequencing (WGS). Results: Five PCR-defined genotypes were distinguished. All strains from HUS cases harboured stx2a and eae, with or without stx1a, while strains from diarrhoea cases carried exclusively stx1a and eae genes. PFGE resolved strains into multiple pulsotypes, compatible with a certain geographic segregation of the cases, and strains were assigned to phylogroup B1 and sequence type (ST) 21. WGS confirmed the results of conventional molecular methods, brought evidence of O26:H11 serotype, and complemented the virulence profiles. Discussion/conclusion: This first description of STEC O26 strains from cases in Romania showed that the isolates belonged to a diverse population. The virulence content of most strains highlighted a high risk for severe outcome in infected patients. Improving the national surveillance strategy for STEC infections in Romania needs to be further considered.

The surveillance of shigellosis is carried out under the auspice of the European Centre for Disease Prevention and Control which requires a reliable laboratory-based surveillance at national level. To date, little information is published about the members of Shigella spp. responsible for Romanian cases of shigellosis which hinders the understanding of the current epidemiology of shigellosis. Consequently, this retrospective study aimed to assess the diversity of virulence profiles displayed by the strains circulating in our region, by using key chromosome- and plasmid-associated markers, and to document the prevalence of pHS-2-like plasmid previously proposed as a potential marker for reactive arthritis.The study focused on 65 Shigella sonnei and 49 Shigella flexneri clinical isolates, originated from local patients, recovered through the national surveillance system in 2009 - 2013. PCR assays were performed for the detection of ipaH, ipaBCD, ial, sen, set1A, set1B, sat, and pic genes, and a PCR-sequencing approach on plasmid preparations was used for identifying pHS-2-specific sequences.Overall, the virulence markers ranged in prevalence from 21% (set1A, set1B, pic) to 100% (ipaH). S. flexneri isolates displayed a higher content of virulence markers than S. sonnei, the most common genotype, detected exclusively in S. flexneri serotype 2a isolates, being defined by the association ipaH+ipaBCD+ial+sen+sat+set1A+ set1B+pic. pHS-2-like plasmids were found in S. flexneri isolates of various serotypes suggesting the potential to trigger postinfection complications in specific subjects.This study provided baseline data regarding the molecular characteristics of the Shigella strains from Romania, useful for defining the picture of shigellosis in our region.

A c c e p t e d M a n u s c r i p t 2 SummaryVerocytotoxin-producing Escherichia coli (VTEC) of serogroup O157 are among the most important causes of severe cases of foodborne disease and outbreaks worldwide. As little is known about the characteristic of these strains in Romania, we aimed to provide reference information on the virulence gene content, phylogenetic background, and genetic diversity of seven autochthonous O157 strains collected during 2016 and 2017 from epidemiologically non-related cases. These strains were typed by a combination of phenotypic and molecular methods routinely used by the national reference laboratory. Additionally, four of them were also subjected to whole-genome sequencing (WGS) and public web-based tools were used to extract information on virulence gene profiles, multilocus sequence types (MLST), and SNPbased phylogenetic relatedness. Molecular typing provided evidence of the circulation of a polyclonal population while distinguishing a cluster of non-sorbitol-fermenting, glucuronidase-negative, phylogenetic group E, MLST 1804 strains, representing lineage II and clade 7, which harbored vtx2c, eae-gamma, and ehxA genes. A good correlation between the routine typing methods and WGS data was observed. Yet SNP-based genotyping provided a higher resolution in depicting the relationships between the O157:H7 strains than that provided by PFGE. This study should be a catalyst for improved laboratory-based surveillance of autochthonous VTEC.

Spondyloarthritis (SpA) is a group of associated chronic systemic inflammatory immune-mediated rheumatic diseases affecting axial and peripheral joints and entheses. The aim of the present study was to identify what parameters are useful to determine in order to better understand the correlation between the disease activity/severity and the microbiological results/immune status against intestinal and/or urogenital pathogens. Microorganisms known to trigger SpA, including Klebsiella spp., Yersinia spp., Salmonella spp., Campylobacter spp . and Chlamydia spp ., were analyzed in various specimens (stool, urine, synovial fluid and serum) collected from 27 randomly selected SpA patients and 26 healthy controls using a combined direct and indirect approach relying on conventional culture technique and nucleic acid-based assays together with serological testing by ELISA. Although Escherichia coli derived from phylogroup A prevailed in the gut microflora of the patients and controls, differences were observed regarding the representatives of the other phylogroups with a higher prevalence of E.coli members of phylogenetic group B1 in the stool specimens of patients. Antibodies against the targeted species were detected in SpA patients and controls, and the serological profiles of the former were more diverse and complex. In conclusion, the detection of anti-bacterial antibodies combined with other specific laboratory investigations should be more extensively used to monitor SpA patients in association with their symptoms and in order to determine and administer more effective therapeutics.