Taxonomic considerations on Aerococcus urinae with proposal of subdivision into Aerococcus urinae, Aerococcus tenax sp. nov., Aerococcus mictus sp. nov., and Aerococcus loyolae sp. nov.

Average nucleotide identity analysis, based on whole genome sequences of 115 strains previously identified as Aerococcus urinae, an emerging uropathogen, discriminates at least six unique genomic taxa. The whole genome analysis affords clearer species boundaries over 16S rRNA gene sequencing and traditional phenotypic approaches for the identification and phylogenetic organization of Aerococcus species. The newly described species can be differentiated by matrix-assisted laser desorption ionization time-of-flight analysis of protein signatures. We propose the emendation of the description of A. urinae (type strain ATCC 51268T = CCUG 34223T=NCFB 2893) and the names of Aerococcus tenax sp. nov. (ATCC TSD-302T = DSM 115700T = CCUG 76531T=NR-58630T), Aerococcus mictus sp. nov. (ATCC TSD-301T = DSM 115699T = CCUG 76532T=NR-58629T), and Aerococcus loyolae sp. nov. (ATCC TSD-300T = DSM 115698T = CCUG 76533T=NR-58628T) for three of the newly identified genomic taxa.

Identification of two abundant Aerococcus urinae cell wall-anchored proteins.

Aerococcus urinae is an emerging pathogen that causes urinary tract infections, bacteremia and infective endocarditis. The mechanisms through which A. urinae cause infection are largely unknown. The aims of this study were to describe the surface proteome of A. urinae and to analyse A. urinae genomes in search for genes encoding surface proteins. Two proteins, denoted Aerococcal surface protein (Asp) 1 and 2, were through the use of mass spectrometry based proteomics found to quantitatively dominate the aerococcal surface. The presence of these proteins on the surface was also shown using ELISA with serum from rabbits immunized with the recombinant Asp. These proteins had a signal sequence in the amino-terminal end and a cell wall-sorting region in the carboxy-terminal end, which contained an LPATG-motif, a hydrophobic domain and a positively charged tail. Twenty-three additional A. urinae genomes were sequenced using Illumina HiSeq technology. Six different variants of asp genes were found (denoted asp1-6). All isolates had either one or two of these asp-genes located in a conserved locus, designated Locus encoding Aerococcal Surface Proteins (LASP). The 25 genomes had in median 13 genes encoding LPXTG-proteins (range 6-24). For other Gram-positive bacteria, cell wall-anchored surface proteins with an LPXTG-motif play a key role for virulence. Thus, it will be of great interest to explore the function of the Asp proteins of A. urinae to establish a better understanding of the molecular mechanisms by which A. urinae cause disease.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry is a sensitive and specific method for identification of aerococci.

Conventional methods for the identification of human-pathogenic aerococci to the species level are not reliable. We show that matrix-assisted laser desorption ionization-time of flight mass spectrometry correctly identifies aerococci to the species level and that it can be used to identify aerococci with high specificity in the diagnostic clinical microbiology laboratory.

Genetic relatedness of Streptococcus dysgalactiae isolates causing recurrent bacteraemia.

Introduction. Streptococcus dysgalactiae subspecies equisimilis (SDSE) is becoming increasingly recognized as an important human pathogen. Recurrent bacteremia with SDSE has been described previously.Aim. The aims of the study were to establish the genetic relatedness of SDSE isolates with emm-type stG643 that had caused recurrent bacteraemia in three patients and to search for signs of horizontal gene transfer of the emm gene in a collection of SDSE stG643 genomes.Hypothesis. Recurring SDSE bacteremia is caused by the same clone in one patient.Methodology. Whole genome sequencing of 22 clinical SDSE stG643 isolates was performed, including three paired blood culture isolates and sixteen isolates from various sites. All assemblies were aligned to a reference assembly and SNPs were extracted. A total of 53 SDSE genomes were downloaded from GenBank. Two phylogenetic trees, including all 75 SDSE isolates, were created. One tree was based on the emm gene only and one tree was based on all variable positions in the genomes.Results. The genomes from the three pairs of SDSE isolates showed high sequence similarity (1-17 SNPs difference between the pairs), whereas the median SNP difference between the 22 isolates in our collection was 1694 (range 1-11257). The paired isolates were retrieved with 7-53 months between episodes. The 22 SDSE isolates from our collection formed a cluster in the phylogenetic tree based on the emm gene, while they were more scattered in the tree based on all variable positions.Conclusions. Our results show that the paired isolates were of the same clonal origin, which in turn supports carriage between bacteraemia episodes. The phylogenetic analysis indicates that horizontal gene transfer of the emm-gene between some of the SDSE isolates has occurred.

Heart transplantation with ABO-identical versus ABO-compatible cardiac grafts: influence on long-term survival.

OBJECTIVES: To compare identical versus compatible, ABO blood group matching effects on rejection and long-term survival after heart transplantation (HT). DESIGN: Data were collected from 196 patients who underwent HT at Lund University Hospital between 1988 and 2008. Cox proportion hazard regression analysis was used to identify factors associated with reduced long-term survival. RESULTS: One hundred and sixty six patients (85%) had an identical ABO blood group match and 30 patients (15%) had a compatible, ABO blood group match. Four non-pharmacological variables reducing overall survival were identified: recipient blood group AB, age >55 years, ischemic time, and year of transplantation. Two pharmacological variables improved overall survival: glucocorticoids and cyclosporine. There was no significant difference in long-term survival between patients with identical blood groups compared to compatible ABO blood group matching. However, there was a trend towards graft failure as cause of death being more common in the compatible ABO group match compared identical blood group match (13% versus 5%, p=0.118). CONCLUSIONS: Six factors associated with overall survival were identified. One of these was related to blood group AB. No significant difference in survival following identical, versus compatible, ABO matching was demonstrated.

A prospective observational treatment study of aerococcal urinary tract infection.

OBJECTIVES: Aerococcus urinae and Aerococcus sanguinicola cause urinary tract infections (UTIs) and antibiotic treatment recommendations are solely based on in vitro findings and limited clinical experience. Our objective was to investigate the effectiveness of different treatment strategies in aerococcal UTI through a prospective observational study. METHODS: Urine samples with aerococci were identified and patients were enrolled. The aerococci were subjected to Etests. Information on clinical symptoms, and the treatment given, was collected. Patients were interviewed after the conclusion of treatment to assess clinical cure and a control urine culture assessed the microbiological cure. RESULTS: Of 31,629 urine samples, 144 grew aerococci and fulfilled the inclusion criteria. 91 patients gave consent and the 72 patients with UTI were assessed for treatment outcome. 53 patients had A. urinae UTI, while 19 had A. sanguinicola UTI. Nitrofurantoin was most commonly prescribed, achieving clinical and microbiological success in 71/76% of cases of A. urinae UTI, and 42/50% of cases of A. sanguinicola UTI. Pivmecillinam achieved success in patients with A. urinae cystitis and ciprofloxacin in patients with pyelonephritis. CONCLUSIONS: Our results support that nitrofurantoin is a valid option for the treatment of cystitis caused by A. urinae.

Epidemiology and antibiotic susceptibility of aerococci in urinary cultures.

In this study, we present population-based data regarding the prevalence of aerococci in clinical urinary samples. During a 3-month period, all aerococcal isolates from urinary samples from 2 clinical microbiology laboratories were collected. We identified 64 Aerococcus urinae isolates and 40 Aerococcus sanguinicola isolates, which correlates with an incidence of 33 cases of aerococcal bacteriuria per 100,000 inhabitants per year. The median age was 83years for all patients with aerococcal bacteriuria, which was significantly higher than for patients with Escherichia coli or Enterococcus faecalis bacteriuria. Sex was almost equally distributed between men and women with aerococcal bacteriuria, whereas females dominated in E. coli bacteriuria. The aerococcal isolates displayed low MICs for ampicillin, cefalotin, mecillinam, and nitrofurantoin. Most A. sanguinicola isolates were resistant to ciprofloxacin, whereas most A. urinae isolates had low MICs. Clinical studies are needed to establish clinical breakpoints and optimal treatment.

Bacteremia with Aerococcus sanguinicola: Case Series with Characterization of Virulence Properties.

BACKGROUND: Since Aerococcus sanguinicola was designated as a species in 2001, only a few cases of bacteremia have been reported. The aim with this study was to describe the clinical presentation of A sanguinicola bacteremia and to determine the antibiotic susceptibility and the capacity of the bacteria to form biofilm and to induce platelet aggregation. METHODS: Isolates of A sanguinicola from blood cultures were retrospectively identified from 2 clinical microbiology laboratories for 2006 to 2012. Species identity was confirmed through sequencing of the 16S rRNA gene. The medical charts of patients were reviewed. The minimum inhibitory concentration (MIC) for relevant antibiotics was determined. Biofilm formation was measured as the amount of crystal violet absorbed. Platelet aggregation was determined by aggregometry. RESULTS: Eleven cases of A sanguinicola bacteremia were identified. All patients were male and the median age was 82 years (range 67-93). Nine patients fulfilled criteria for severe sepsis, and 2 patients died at hospital. Two patients were diagnosed with infective endocarditis. Most patients had underlying urinary tract diseases or an indwelling urinary tract catheter. Five patients suffered from dementia. None of the patients was treated with immunosuppressive medications. The MIC values of the isolates were in line with previous reports, with low MICs for penicillin, cefotaxime, and vancomycin. All 11 isolates produced biofilms but not all could induce platelet aggregation. CONCLUSIONS: A sanguinicola can cause severe infections in elderly men with urinary tract abnormalities and the bacteria possess potential virulence mechanisms.