Detection of clinically relevant antimicrobial resistance determinants in warm-blooded marine animals in Livingston Island (South Shetland Islands, Antarctica): A field-based molecular genetics study.

Molecular genetic studies of stools were performed to assess the spread of some clinically relevant antimicrobial resistance determinants (ARD) in a gentoo penguin (Pygoscelis papua) and an Antarctic fur seal (Arctocephalus gazella) on Livingston Island. Glycopeptide resistance genes (vanA/vanD and vanB) were detected in both fecal samples, while the penguin's one was also mecA-positive and blaNDM-positive. Because of the remoteness and the isolation of the sampling locations, the carriage of vancomycin-resistant Enterococcus spp., methicillin-resistant Staphylococcus aureus, and NDM-producing Enterobacterales or other gram-negative bacilli suggested an ocean pollution with antibiotic resistant bacteria (ARB). Additionally, due to the type of ARD we detected, our results are alarming, and they cannot be explained only with agricultural and/or aquacultural pollution. Even though the current study is a preliminary one, it also demonstrates the potential of the field genetics analyses carried out with minimal equipment as a reliable monitoring tool for pollution with ARB.

Microbiological diagnostics of bloodstream infections in Europe-an ESGBIES survey.

OBJECTIVES: High-quality diagnosis of bloodstream infections (BSI) is important for successful patient management. As knowledge on current practices of microbiological BSI diagnostics is limited, this project aimed to assess its current state in European microbiological laboratories. METHODS: We performed an online questionnaire-based cross-sectional survey comprising 34 questions on practices of microbiological BSI diagnostics. The ESCMID Study Group for Bloodstream Infections, Endocarditis and Sepsis (ESGBIES) was the primary platform to engage national coordinators who recruited laboratories within their countries. RESULTS: Responses were received from 209 laboratories in 25 European countries. Although 32.5% (68/209) of laboratories only used the classical processing of positive blood cultures (BC), two-thirds applied rapid technologies. Of laboratories that provided data, 42.2% (78/185) were able to start incubating BC in automated BC incubators around-the-clock, and only 13% (25/192) had established a 24-h service to start immediate processing of positive BC. Only 4.7% (9/190) of laboratories validated and transmitted the results of identification and antimicrobial susceptibility testing (AST) of BC pathogens to clinicians 24 h/day. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry from briefly incubated sub-cultures on solid media was the most commonly used approach to rapid pathogen identification from positive BC, and direct disc diffusion was the most common rapid AST method from positive BC. CONCLUSIONS: Laboratories have started to implement novel technologies for rapid identification and AST for positive BC. However, progress is severely compromised by limited operating hours such that current practice of BC diagnostics in Europe complies only partly with the requirements for optimal BSI management.

[Risk factors for bacterial vaginosis - a 3-year study in four medical centers].

BACKGROUND: Bacterial vaginosis (BV) is the most common infection of the lower genital tract among women of reproductive age. The purpose of this study was to determine the frequency of BV and the important etiological agent Gardnerella vaginalis in Bulgarian patients of different age groups, as well as, the risk factors forBV METHODS: One thousand five hundred and twenty-three (1523) women aged 16 to 45 years without previous antimicrobial therapy were included in this study. The methods used were: scoring of Gram staining of vaginal smear and polymerase chain reaction (PCR) assay for G. vaginalis. RESULTS: Positive for BV according to the microscopic examination criteria were 47.80% from the 1523 tested women. In 88.87% from the positive samples G. vaginalis was detected using PCR, thus demonstrating high comparability of the both methods and the leading role of G. vaginalis. The most affected with BV is the age group of 21-25 years (33.21% of all subjects with BV). The most important detected risk factors were: smoking (more than 55% with BV), single marital status (only 15% with BV are married), more than I sexual partner (more than 36% had changed the sexual partner), early onset of sexual activity (75% of B V asocciated subjects started their sexual activity by the age 15-18 years). CONCLUSIONS: The established early age for aquiring BV among Bulgarian women is very important and alarming sign. This is the first study on the etiological role of G. vaginalis and on the risk factors for BV in Bulgaria.

Antimicrobial activity of tobramycin against respiratory cystic fibrosis Pseudomonas aeruginosa isolates from Bulgaria.

Tobramycin solution for inhalation (TSI) (Novartis pharmaceuticals) is indicated as chronic suppressive treatment for cystic fibrosis (CF) patients aged 6 years and older who are chronically infected by Pseudomonas aeruginosa . Inhaled administration of tobramycin assures high concentrations in the lungs of CF patients, improving the therapeutic ratio over that of parenteral tobramycin levels. Clinical and laboratory Standards institute (CLSI) breakpoints only consider parenteral levels and do not take into account these high antimicrobial concentrations. Therefore, the Spanish meNSURA Group has defined specific values for inhaled tobramycin when testing CF P. aeruginosa isolates (susceptible: minimal inhibitory concentration (MIC) ≤ 64 µg/ml; resistant: ≥ 128 µg/ml). In this study the antimicrobial activity of tobramycin against 120 respiratory CF P. aeruginosa isolates was determined by high-range etest strips (LIOFILCHEM). Applying MENSURA breakpoints, 95% of the strains were categorized as susceptible. With CLSI breakpoints, the susceptibility rate decreased to 92.5%. The activity against non-mucoid P. aeruginosa was higher than that against mucoid isolates (MIC(50)=0.75 and MIC(90)=2 µg/ml vs. MIC(50)=1 and MIC(90)=4 µg/ml). The isolates obtained from patients untreated with TSI were more susceptible to the drug than those from patients receiving maintenance therapy with TSI (MIC(50)=0.75 and MIC(90) =1.5 µg/ml vs. MIC(50)=1.5 and MIC(90)=6 µg/ml). The isolates from patients with long-term P. aeruginosa colonization (over 5 years) revealed the highest tobramycin MICs (MIC(50)=1.00 and MIC(90)>1024 µg/ml). In conclusion, tobramycin has excellent in vitro activity against the studied CF isolates. Some factors such as isolate morphotype, pre-administration of TSI and duration of colonization influence its activity. Whenever TSI is considered for therapy, the CF P. aeruginosa strains categorized as intermediate or resistant to tobramycin according to the CLSI criteria should be recategorized by using the MENSURA interpretive criteria.

Widespread detection of VEB-1-type extended-spectrum beta-lactamases among nosocomial ceftazidime-resistant Pseudomonas aeruginosa isolates in Sofia, Bulgaria.

A total of 132 ceftazidime-resistant clinical isolates of Pseudomonas aeruginosa were collected during 2001-2005 from 5 university hospitals in Sofia, Bulgaria to assess the current levels of antimicrobial susceptibility and to evaluate resistance mechanisms to beta-lactams. Antimicrobial susceptibilities were detected by a disk diffusion method and E-test. Polymerase chain reaction amplification and sequencing of bla(VEB-1 )and bla(PER-1 )were performed. The antibiotic resistance rates were: to piperacillin 90.2%, piperacillin/tazobactam 52.3%, ceftazidime 94.7%, cefepime 88.6%, cefpirome 98.5%, aztreonam 85.6%, imipenem 66.6%, meropenem 63.6%, amikacin 81.1%, gentamicin 84.8%, tobramycin 89.4%, netilmicin 57.6%, ciprofloxacin 83.4%. Structural genes for VEB-1 extended-spectrum beta -lactamases (ESBLs) were found in 75 (56.8%) of the isolates. PER-1 ESBLs were not detected. The VEB-1-producing strains were more resistant than VEB-1 non-producers to amikacin, gentamicin, tobramycin and ciprofloxacin ( P<0.001). VEB-1 appears to have a significant presence among ceftazidime-resistant P. aeruginosa isolates from Sofia.