Purpose
Data on the molecular epidemiology of methicillin-resistant
Staphylococcus aureus
isolates from patients with bacteraemia in Slovenia are lacking. The aim of this study was to phenotypically ...and genotypically investigate 82 MRSA strains isolated from patients with bloodstream infections in central Slovenia between 2019 and 2022.
Methods
Whole-genome sequencing of selected strains was performed to characterize the strains based on sequence typing, antimicrobial resistance, toxin, and virulence factors genes.
Results
Most MRSA carried SCC
mec
II (63.4%), followed by SCC
mec
IV (34.1%) and SCC
mec
V (2.5%). A high proportion of strains belonging to the ST225 lineage (45.1%) was observed, followed by ST97 (18.3%), ST2883 (15.9%), ST22 (9.8%), ST5 (3.7%), and the ST1, ST398 and ST45 lineages (2.4% each). Sixteen different
spa
types were identified, predominantly ST225-t003 (31.7%), ST97-t359 (15.9%), and ST2883-t4336 (14.6%). None of the strains carried Panton-Valentine leukocidin, exfoliative toxins, or toxic shock toxin. All MRSA strains were susceptible to linezolid, rifampicin, vancomycin, teicoplanin, and trimethoprim-sulfamethoxazole. MRSA strains were resistant to erythromycin, clindamycin, tetracycline and gentamicin, with a frequency of 74.4%, 74.4%, 8.5%, and 1.2%, respectively.
Conclusion
This study demonstrates that bacteraemia in central Slovenia is caused by diverse MRSA lineages. Identification of newly emerged lineages should be followed in the future to detect changes in the molecular epidemiology of MRSA in our country.
The identification of patients infected and/or colonised by methicillin resistant
(MRSA) is necessary for the timely introduction of measures for infection control. We compared the diagnostic ...efficacy of combinations of MRSA surveillance swabs routinely taken by health institutions in the country.
All surveillance samples, which were sent for a microbiological analysis to detect MRSA with the culture method in 2014, in the three departments for medical microbiology of the National Laboratory for Health, Environment and Food, were included in this study.
Among 65,251 surveillance cultures from 13,274 persons, 1,233 (2.1%) were positive (490 positive persons). Prevailing positive surveillance cultures were throat swabs (31.3%), followed by nose swab (31.2%), skin swab (18.9%), perineum (16.4%) and wound swabs (1.4%). The contribution of other samples, such as aspirate, urine and excreta, was under 1%. We found no statistically significant differences in the frequency of detection of a positive patient, if the combination of samples NTS (nose, throat, skin) or NTP (nose, throat, perineum) was analysed. However, statistically significant differences were confirmed when any of the anatomic sites would be omitted from the sets of NTP and NTS (chi square; p<0.01). Adding additional samples resulted in only 24 additional positive patients (4.9%).
The results indicate that increasing the number of surveillance cultures above three does not add much to the sensitivity of MRSA surveillance, the exception could be wound. The swabs from the perineum and from the skin are exchangeable.
Although the distinction between the Community-Associated-Methicillin-Resistant
(CA-MRSA) and Hospital-Associated-Methicillin-Resistant
(HA-MRSA) has blurred in recent years, the CA-MRSA is an ...important group because of its potential to cause fulminant and severe infections. Its importance has further increased with the emergence of Livestock-Associated-Methicillin-Resistant
(LA-MRSA).
In the present study we analysed clonal distributions and virulence factors in presumptive CA-MRSA isolated from January 2014 to December 2015 and compared the results with our previous study from 2010. Phenotypic definition for presumptive CA-MRSA was based on resistance to cefoxitin and oxacillin and susceptibility to at least two of the following four antibiotics: ciprofloxacin, erythromycin, clindamycin and gentamicin.
In 2014 and 2015 altogether 304 MRSA isolates fulfilled our screening phenotypic definition, 45 isolates were cultivated from clinical specimens and 259 from screening specimens. Sequence types ST398, LA-MRSA and
C MRSA increased significantly in 2015 compared to 2010 (p-value <0.05) and were spread over Slovenia.
The clonal distribution of presumptive CA-MRSA has changed within the study period in Slovenia. In 2015 the most frequent clone among clinical and screening specimens was a pig-associated clone, ST398, but the number of confirmed ST398 infections remains low. While previously ST398 and
C positive MRSA strains were geographically limited, they have spread throughout the country since 2010.
Background: Acute uncomplicated cystitis in women is a common reason for antibiotic treatment in the outpatient clinic of family doctors. Microbiological diagnostics is not indicated; treatment is ...generally empirical and based on guidelines and recommendations. Therefore, we do not have representative data on the incidence and sensitivity of pathogens from regular laboratory work. Special targeted research is needed.
Methods: We present the results of the prospective research conducted in four regional laboratories of the Centre for Medical Microbiology NLZOH between 15 September 2017 and 31 December 2019. We included 110 female patients with suspected acute uncomplicated cystitis who completed a questionnaire and submitted urine samples for microbiological examination.
Results: The result of urine examination was positive in 79 patients (71.8%). Among all isolates, the most common bacterium was Escherichia coli (74%), followed by Staphylococcus saprophyticus (10%), and others. The susceptibility of E. coli to trimethoprim with sulfamethoxazole was 85.5%, to nitrofurantoin 98.4%, and to fosfomycin 100%. S. saprophyticus isolates were susceptible to the tested antibiotics except for fosfomycin, against which the bacterium is intrinsically resistant. The total susceptibility of all isolates was 85.8% - 88.2% for trimethoprim with sulfamethoxazole, 88.1% - 95.2% for nitrofurantoin, 73.8% - 80.8% for fosfomycin, 72.6% for ampicillin and amoxicillin, 97.7% for cefadroxil, and 98.9% for amoxicillin with clavulanate acid. A comparison with the results of monitoring bacterial resistance as part of the regular work of microbiological laboratories shows significant differences for most of the tested antibiotics.
Conclusion: The research results show a relatively good susceptibility of the causative agents of acute uncomplicated cystitis to oral antibiotics, the only exception being amoxicillin.
Bacterial vaginosis is of clinical interest because of its possible causal relationship with complications during pregnancy, postpartum, and complications after surgery.
Gram stain for clue cells and ...Gardnerella vaginalis culture methods were evaluated retrospectively in a microbiological medical laboratory for the first half of 2015. We were interested in the proportion of G. vaginalis bacteria isolated from genital samples, correlation with Gram-staining presence of clue cells, referral clinical diagnosis, and pregnancy.
In the first half of 2015 we received 358 vaginal specimens; 82% of them had a referral clinical diagnosis of colpitis, cervicitis, or vaginal discharge; 40% were pregnant women. G. vaginalis was isolated from 14% of vaginal specimens, and 52% of these came from pregnant patients. Gram stain clue cells and isolation of G. vaginalis matched in 86%.
For diagnosing bacterial vaginosis in clinical practice, standard clinical criteria, Gram staining of vaginal discharge smear, and/or isolation of G. vaginalis are used. Isolation of G. vaginalis without clue cells is reported only in cases in which bacterial growth is predominant. The results of our studies confirm that isolating G. vaginalis helps confirm the diagnosis of bacterial vaginosis.
Aim
To characterize methicillin-resistant S. aureus (MRSA) strains phenotypically and genotypically and to determine their clonal affiliation, representation and antibiotic resistance profile.
...Methods
A total of 62 randomly selected MRSA isolates of different clinical samples collected from 2009 to 2017 were phenotypically and genotypically analysed. Phenotypic analyses were performed by standard microbiological procedures, and using VITEK 2/AES instrument as well as MALDI-TOF (matrix-assisted laser desorption/ionization) technology. Genotypic characterization included spa, MLST (multilocus sequence typing) and SCCmec typing, and detection of the Panton-Valentine leukocidin (PVL) and other enterotoxin encoding genes.
Results
The largest number of isolates, 21 (33.87%) belonged to ST228-MRSA-I, spa type t041, t1003 and t001. Other major
clones were: ST239-MRSA-III, spa type t037 and t030 (27.41%); ST8-MRSA-IV, spa type t008 and t121 (12.9%); ST247-MRSA-I, spa type t051 (4.83%). PVL was detected in 10 isolates (SCCmec IV/V). During 2009 and 2010 the most frequent MRSA strain was South German clone, ST228-MRSA-I (80% and 90%, respectively), while in later years it was replaced with Brazilian-Hungarian clone ST239-MRSA-III (75% in 2015 and 2016). The South German clone, spa type t041 in 90.48% of cases was resistant to clindamycin, ciprofloxacin, erythromycin, cefoxitin, gentamicin, kanamycin, tobramycin and penicillin, while 70.58% samples of the Brazilian-Hungarian clone spa type t037 were additionally resistant to tetracycline and rifampicin.
Conclusion
This research can supplement the existing knowledge about the clonal distribution of MRSA in Bosnia and Herzegovina
and their sensitivity to antibiotics in order to improve the national control of these infections
Abstract
Introduction
According to the existing literature, a heterogeneous sequence type (ST) or clones of community-associated methicillin-resistant
Staphylococcus aureus
(CA-MRSA) circulate in ...Europe. In Europe, the European clone that belongs to sequence type ST80 is predominant.
Methods
The aim of the study was to investigate the phenotypic and genotypic characteristics and epidemiological data of CA-MRSA ST80 and its occurrence in Slovenia. We retrospectively analyzed those CA-MRSA isolates that were isolated during microbiological procedures in microbiological laboratories between 2006 and 2013. Only CA-MRSA isolates from the national collection of CA-MRSA strains that belonged to ST80 (European clone) were analyzed. We determined the Pantone-Valentine leukocidin (PVL),
mec
A genes, exfoliative toxin genes and type of staphylococcal cassette chromosome (SCC
mec
) by polymerase chain reaction (PCR). We determined also spa type and sequence type.
Results
ST80 was confirmed in only 2 (0.5%) out of 385 CA-MRSA isolates, collected in a national collection of CAMRSA. Both isolates were positive for the PVL genes,
mec
A gene, exfoliative toxin type D gene and SCC
mec
IV. One CA-MRSA isolate was confirmed in a wound swab taken from a 47-year-old male, and the second was isolated from blood cultures of a 69-year-old female. No epidemiological connections between them were found.
Conclusions
In Slovenia CA-MRSA infections caused by ST80 are rare. In the future, it is necessary that a surveillance study of CA-MRSA at the national level continues and CA-MRSA be considered as a public health threat.
Methicillin-resistant Staphylococcus aureus (MRSA) is a major human pathogen and an important cause of hospitalassociated (HA-MRSA) infections. MRSA infections significantly increase morbidity and ...mortality, affect the increased use of antibiotics and the cost of treatment. During the last decade MRSA has emerged as a significant pathogen also in the community (community-associated; CA-MRSA). In recent years, livestock has been proven to be a source of human infections with the MRSA sequence type (ST) 398 (livestock-associated; LA-MRSA). During the year 2010 all the regional microbiological laboratories took part in the task of monitoring CA-MRSA infections in Slovenia. We included all patients harbouring a MRSA strain that was susceptible to at least two of the following four antibiotics: ciprofloxacin, erythromycin, clindamycin or gentamicin. Altogether we collected 151 MRSA isolates of which 15 (9.9%) belonged to a spa type known to be associated with the clone ST398 respectively. Among them 12 isolates belonged to spa type t011, 2 isolates to t034 and 1 isolate to t108. We found the staphylococcal cassette chromosome - SCCmec type IV or V, and regulatory genes - agr type I. None of the isolates were positive for Panton - Valentine leukocidin (PVL), the toxic shock syndrome toxin (tst) and leukocidin LukM. All MRSA isolates were resistant to tetracycline and penicillin. Some of them were also resistant to erythromycin and clindamycin. Most of the LA-MRSA ST398 were isolated from screening specimens of patients from Murska Sobota and Maribor, which are the most important agricultural regions with intensive livestock breeding. Evidence of the presence of LA-MRSA in humans requires a close cooperation of human and veterinary microbiologists. Our goal is to find the epidemiological relation between human and animal hosts, to obtain information on the phenotypic and genotypic characteristics and monitor infections caused by LA - MRSA strains.
Background: In October 2010 an outbreak of staphylococcal food poisoning occurred in an elementary school with a kindergarten in Gorenjska. Methods: Using a questionnaire we performed a retrospective ...cohortanalytical epidemiological study. We calculated attack rate (AR) and relative risk for each food item. In patients we cultivated stools and vomit. We performed an onsite audit where the food samples, environmental samples, and swabs of the hands of the cooks were taken. We performed medical examination of those employed in the kitchen where specimens were taken for microbiological examination. Staphylococcal enterotoxins were detected with agglutination test SET RPLA (Oxoid) and genotypes determined by the pulsed-field gel electrophoresis (PFGE). Results: Out of 374 exposed 73 subjects got ill. Attack rate in the outbreak was 19,5 %. The most frequent symptoms were vomiting (87,7 %), stomach ache (75,3 %), diarrhea (64,2 %) and nausea ( 59,3 %). The highest relative risk (RR) were found for the meatloaf (RR= 24,2 (95 % CI 12,1-48,5; p<0,001)) and potato salad (RR= 19,4 (95 % CI 10,7 - 35,2; p<0,001)). Enterotoxin A producing S. aureus was isolated from hand of a cook, from potato salad and meatlof, from vomit and stools of patients. These strains had the same antibiotic sensitivity and were genetically closely related (96,3 %). Conclusions: In the article an optimal team approaches of a community epidemiologist and microbiologist in the occasion of outbreak are described. We confirmed epidemiologically related isolates and determined the importance of individual methods.
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