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This study aimed to describe the virulence gene content and antibiotic resistance profiles as well as the genetic relatedness of shigella strains originated from romanian patients, collected between 2016-2018 through the national diarrheal disease surveillance program. A total of 60 Shigella sonnei strains and 26 S. flexneri strains were tested against a set of 16 antimicrobials, screened by PCR for several genes involved in the disease pathogenesis (ipah, ipaBCD, ial, sen, set1A, set1B, sat and pic genes), and typed by pulsed-field gel electrophoresis (PFGE) to make inferences about their genetic relatedness. Overall, the resistance most commonly acquired by the autochthonous Shigella strains investigated was against sulfonamide compounds (58/86 strains), trimethoprim (46/86 strains) or combination of trimethoprim with sulfamethoxazole (44/86 strains), and ampicillin (41/86 strains). Strains with resistance to extended spectrum cephalosporins (12/86 strains) were detected, all extended spectrum beta-lactamase producers, as well as strains that displayed resistance to nalidixic acid (3/86 strains) but not to ciprofloxacin. the virulence markers targeted ranged in prevalence from 6% (set1A, set1B, pic) to 100% (ipah). The S. flexneri strains displayed a richer virulence gene content than s. sonnei strains. the most complex virulence genotype, represented by the association of ipah, ipaBCD, ial, sen, sat, set1A, set1B, and pic genes was found in S. flexneri serotype 2a strains. The PFGE-based molecular typing allowed the detection of several clusters of strains displaying band-based profiles with at least 85% similarity. Eight such clusters were identified among the s. sonnei strains and four clusters among the S. flexneri strains. Continued laboratory-based surveillance is essential to generate data that help to draw the picture of the public health problem represented by shigellosis in Romania. [ABSTRACT FROM AUTHOR]

Introduction: Due to the drawbacks of traditional serotyping, many laboratories tend to adopt in their routine work rapid and accurate molecular alternatives for serotyping. Objectives: This study was designed to assess a PCR-based approach for Shigella flexneri serotyping that was chosen to be used at reference laboratory level in order to strengthen microbiological surveillance of autochthonous strains for public health purposes. Methods: A total of 113 S. flexneri strains (19 reference strains and 94 strains isolated between 2007-2019 from autochthonous diarrhea cases) previously serotyped by slide agglutination with commercially available antisera were used to validate the PCR-based molecular serotyping protocol. The molecular approach selected combined multiplex and simplex PCR assays and allowed the identification of 14 most common S. flexneri serotypes based on the amplification patterns generated with primers targeting O-antigen synthesis and modification genes. Results: The PCR-based system of serotyping provided the possibility to assign all studied strains, including those that reacted only with type-specific antisera (11 strains), to distinctive serotypes. However, while conventional and molecular serotyping results correlated perfectly for the reference strains, disconcordances between the phenotypic and genotypic data were observed for 12% of the autochthonous isolates which were phenotypically assigned to F4b, F4c, FX, and FY serotypes. Conclusions: The molecular protocol will be implemented in the workflow of the reference laboratory to surpass the limitations of traditional serotyping and enhance surveillance of Shigella infections. Moreover, analysis at whole genome level resolution will be considered in the future for cases requiring a more detailed epidemiological investigation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]