Staphylococcus aureus of phage type 187 isolated from people occurred to be a genes carrier of eneterotoxin C and toxic shock syndrome toxin-1 (TSST-1).

The aim of this study was to examine the genotype properties of Staphylococcus aureus of phage type 187 strains that constitute a separate group among the strains of S. aureus. Sixteen strains were collected from the hospital patients (n=12) and the healthy carriers (n=4) in 13 medical centres in Poland during 1991 and 2005. Biotyping, antibiotic susceptibility, phage typing, detection the genes of enterotoxins and toxic shock syndrome toxin, genotyping of chromosomal DNA by pulsed-field gel electrophoresis (PFGE), also amplification and restriction analysis of the coagulase (coa) and the protein A genes (spa) (PCR/restriction fragment length polymorphism (RFLP)) was tested. The results of this study showed that all staphylococcus of phage type 187 belonged to the human biotype (A) and appeared to be sensitive to all of the tested antibiotics, including methicillin (MSSA). Finding out the toxin genes showed that almost all of them (93.8%) had the enterotoxin C gene (sec) and TSST-1 gene (tst). The PFGE typing proved that the phage type 187 strains (except for one) constitute one PFGE type. These results and the identical restriction patterns in the PCR/RFLP method, also the same biotype, sensitivity to antibiotics and the presence genes of the same type of toxins confirmed that the phage type 187 strains constitute one clone within our country. Additionally, the fact that almost all of them have the enterotoxin genes and tst gene allows to consider them the strains of potentially high virulence.

[Reduced susceptibility to triclosan in methicillin-resistant Staphylococcus aureus]. / Obnizona wrazliwosc na triklosan metycylinoopornych gronkowców zlocistych.

Susceptibility to triclosan in Staphylococcus aureus was determined. The study was carried out on 200 strains, including 100 resistant (MRSA) and 100 susceptibile (MSSA) to methicillin. The examined strains were isolated from varied clinical samples and patients in 18 medical centers, in majority from hospitals in the region of Gdansk. The susceptibility was estimated by the MIC (minimal inhibitory concentration) using dilution test in Mueller-Hinton agar. The antimicrobial resistance patterns were determined, including resistance to methicillin and mupirocin. The most of MRSA strains (62%) demonstrated reduced susceptibility to triclosan (MIC 2mg/L), while 93% of MSSA strains were highly sensitive to this antibacterial agent (MIC 0,031mg/L). The majority (66,1%) of MRSA strains with reduced susceptibility to triclosan demonstrated the same antimicrobial resistance pattern. Reduced susceptibility to triclosan was observed in 8 from 9 high - level mupirocin resistant strains, but the most of MRSA strains with reduced triclosan susceptibility (91,5%) were found among fusidic acid resistant strains.

[Presence of enterotoxin C and toxic shock syndrome toxin--1 (TSST-1) genes in population of Staphylococcus aureus phage type 187]. / Obecnosc genów enterotoksyny C i toksyny wstrzasu toksycznego--1 (TSST-1) w populacji Staphylococcus aureus fagotypu 187.

The aim of this study was to examine whether Staphylococcus aureus of phage type 187 possess the genes of enterotoxins and toxic shock syndrom toxin. Sixteen phage type 187 strains were isolated from the hospital patients (12) and the carriers (4) in twelve medical centres in Poland during 1991 and 2005. Biotyping, phage typing, antibiotic susceptibility, detection of the genes of enterotoxins (sea--sed) and toxic shock syndrome toxin (tst) was tested. The results of this study showed that all staphylococci of phage type 187 belonged to the human biotype (A) and appeared to be sensitive to all of the tested antibiotics, including methicillin (MSSA). Almost all of them (93.8%) had the enterotoxin C gene and TSST-1 gene. This fact allows to consider them the strains of potentially high virulence.

[Phage types of Staphylococcus aureus isolated in Poland in 1999-2004]. / Fagotypy Staphylococcus aureus izolowane w Polsce w latach 1999-2004.

The aim of study was to determine how population of Staphylococcus aureus strains from hospital environment forms itself with respect to phage types and frequency of their occurrence recently. 1157 S. aureus isolated from various clinical materials between 1999 and 2004 have been investigated. The basic set of phages and the three additional ones were used for typing strains according to Blair and Williams method. The results proved that population of S. aureus strains isolated from hospital environment differentiates with respect to phage types, like in the past. Strains belonging to phage group II dominates among them, but a considerable increase in number of phage group III have been remarked lately. A differential value of an individual phages from basic set remains on their usual level for years.

[Protein a production in coagulase-negative or clumping factor (CF)-negative isolates of Staphylococcus aureus]. / Wytwarzanie bialka a przez izolaty Staphylococcus aureus nie wytwarzajace koagulazy lub czynnika zlepnego (CF).

The aim of study was to determine a production of proteinA in coagulase-negative Staphylococcus aureus (CNSA) or CF-negative S. aureus (CFNSA) strains. 59 CNSA and 18 CFNSA strains were isolated between 1997 and 2003 from different clinical specimens. The Protein A production was determined by immunoblotting method. The presence of protein A gene (spa) was investigated using the polymerase chain reaction (PCR). Two sets of phages and RFLP (Restriction Fragment Lenth Polymorphism) of coa gene method were used for typing strains. The results proved that the lack of ability of protein A production occurs more frequently in protein A-negative CFNSA strains with compare to the CNSA, which are protein A-positive for the majority of strains. Deficiencies of protein A, doesn't seem to be caused by the loss of spa gene. Protein A-negative CFNSA strains have phagotypes, RFLP and antibiotic resistant patterns which differ them from protein A-negative CNSA strains. Almost all of protein A-negative CFNSA and CNSA strains are resistant to methicillin.

[Flow cytometric analysys of Enterococcus faecalis pAD1(-) strains response to cAD1 pheromone]. / Zróznicowanie agregacji komórek Enterococcus faecalis pAD1(-) pod wplywem feromonu cAD1.

Conjugative plasmids transfer in Enterococcus faecalis is inducted by sex pheromones. The pheromone is excreted by recipient cells and induces expression of aggregation protein AS in donor cells. This protein is involved in formation of matting aggregates. Use of flow cytometry and anti-As monoclonal antibodies allowed collect of interesting data pheromone response. However, according to our knowledge, no study focused on unspecific influence on particular pheromone for plasmid-free recipient strains. Six pAD1 (-) and tree pAD1 (+) Enterococcus faecalis stains were cultivated for 18h in BHI, with and without cAD1 pheromone (Sigma, Germany), respectively. The bacteria were washed, stained with carboksyfluorescein (FCDA, and analyzed by flow cytometry in FACS BD scan cytometr. Relative fluorescence and size of aggregation was used to compare influence on particular strains. Surprisingly, the results shows divergence in fluorescence, size of aggregates and degree of correlation between fluorescence of aggregates and their sizes among pAD1(-) strains, allowing for distinguish of two groups. Three of studied strains have higher fluorescence than pAD (+) stains. Correlation between fluorescence and size of aggregates, significant higher than in pAD1(+) stains, decrease from r = 0.88 to r=0.74 in reaction to cAD1. The strains if other group fluorize with lower intensity than pAD1 (+). Furthermore, 30.4% pAD1 (-) of second group have no detectable fluorescence. In contrast to pAD1 (-) ) strains of the first group and pA1 (+) strains, low (r=0.55) correlation between fluorescence and size of aggregates of group II increase up to r=0.74 after incubation with cAD1 pheromone. Previous study of these pAD1 (-) strains, currently assigned to group II, shown their low frequency of collecting aph2" gene encoded on other conjugative plasmid, pMG. According to these results, such flow cytometric analysis may be used to predict ability of strain to collect unrelated conjugative plasmid.

[Disappearance of differences in antibiotic resistance of Staphylococcus aureus strains isolated in hospitals and in general practice]. / Zanikanie róznic w lekoopornosci szczepów Staphylococcus aureus izolowanych od pacjentów hospitalizowanycyh i leczonych ambulatoryjnie.

There is general opinion that Staphylococcus aureus strains isolated in hospitals are more frequently resistant to antibiotics than community strains, however, the increasing resemblance between hospital and community strains has been recently reported. The aim of the study was to compare the antibiotic resistance and phage-type pattern of S. aureus strains isolated from patients treated either in hospitals or in general practice in northern part of Poland. The study was conducted on 771 S. aureus strains isolated from different specimens. Phage typing was performed according to the method of Blair and Williams. The drug susceptibility was determined by the disc-diffusion method. There were no significant differences in antibiotic resistance or phage-type pattern when hospital and community methicillin-sensitive S. aureus (MSSA) strains were compared. The most MSSA were resistant to penicillin (84.6% and 82.1% respectively) and doxycycline (49.3% and 50.4% respectively) whereas they were rarely resistant to other antibiotics. The predominance of phage group II was found in both hospitals (28.0%) and general practice (29.9%). Phage group III, usually associated with hospitals, occurred in small percentage (12.9% and 9.4% respectively) while to this group predominantly (76.6%) multiresistant methicillin resistant S. aureus (MRSA) isolated in hospitals belonged. These results suggest, that there is only slight difference in antibiotic resistance between hospital and community S. aureus strains. Antibiotic resistance pattern mainly results from frequency of appearance of MRSA, mostly occurring in hospitals.

[Occurrence of high-level mupirocin resistant coagulase-negative staphylococci in hospitals and their characterization]. / Wystepowanie i charakterystyka gronkowców koagulazo-ujemnych opornych na mupirocyne.

Occurrence of high-level mupirocin resistant coagulase-negative staphylococci in 5 hospitals in the region of Gdansk was determined. The study was carried out on 192 staphylococcal strains isolated from patients and medical staff. The mupirocin resistant strains were detected by 5 microgram mupirocin disc. The minimal inhibitory concentration (MIC) for mupirocin was estimated by E-tests. The mupirocin resistant strains were characterised by antibiotic sensitivity, including MIC for glycopeptides and oxacillin. Biotypes of resistant strains were also determined. Eight high-level mupirocin resistant strains (4.7%) were found. Only one strain expressed low-level resistance. All but one of high-level mupirocin resistant strains were resistant to methicillin. Six of them belonged to the S. epidermidis species but differences in the biotypes and antibiotic resistance patterns of these strains suggest they did not have a common origin.

[Application of PCR-fingerprinting reactions for typing group B Streptococci using ERIC-1 and ERIC-2]. / Zastosowanie reakcji PCR-fingerprinting do typowania paciorkowców grupy B z wykorzystaniem starterów ERIC-1 i ERIC-2.

Usefulness of PCR--fingerprinting technic using ERIC 1 and ERIC 2 primers for epidemiologic analysis of Group B Streptococci was assessed. 120 strains isolated from various clinical samples were tested. Amplification reactions were carried out in automatic termocycler Poly Gen using two primers ERIC 1 and ERIC 2. Products of the amplification were subjected to horizontal electrophoresis in 1.5% agarose gel. Using primer ERIC 1 six DNA patterns were found. Patterns B and C were the most often encountered, containing 34.2% and 35.0% of strains. Less often pattern D occurred including 12.5% of strains. In case of primer ERIC 2 lower differentiation has been achieved, because only four DNA patterns has been found, three of them occurred with similar frequency 29.2% to 33.3%. Taking into consideration the patterns gained by means of two primers thirteen genotypes have been identified. Dominated two patterns CD and BC, comprising 30.0% and 24.1% Group B Streptococci. More rarely DA and BA occurred (10.8% and 8.3%). Remaining patterns make 0.8% to 6.7% strains. Among reference Group B Streptococci strains belonging to nine serologic types using two primers various genetic patterns were obtained.

[Characteristics of defective strains of methicillin resistance Staphylococcus aureus that do not produce coagulase or CF]. / Charakterystyka defektywnych szczepów MRSA nie wytwarzajacych koagulazy lub czynnika CF.

The aim of the study was to estimate frequency of coagulase-negative and CF-negative strains among methicillin-resistant Staphylococcus aureus (MRSA) and to assess their homogeneicity in respect of genotype, phagotype and drug resistance pattern. A total of 186 MRSA strains collected from different hospitals in Gdansk region were studied. Gens: nuc, mecA, and coa were identified by PCR method. The coagulase tube test for staphylocoagulase and the slide test for clumping factor were used. Coagulase-negative and CF-negative MRSA strains were confirmed by PCR-RFLP method of coa gene; phage typing and drug resistance pattern were evaluated by disc diffusion test. The results of the study showed low frequency of both coagulase-negative and CF-negative MRSA strains (7.25% and 3.76% respectively). Among MRSA population tested the simultaneous occurrence of the strains lacking coagulase and clumping factor was not observed. All coagulase negative MRSA had coagulase gene (coa) and differed from CF-negative strains in respect of coa gene.

[Methicillin-resistant Staphylococcus aureus (MRSA) with high resistance to mupirocin in hospitals of the Gdansk region]. / Metycylinooporne gronkowce zlociste (MRSA) o wysokiej opornosci na mupirocyne w szpitalach regionu gdanskiego.

Occurrence of high-level mupirocin resistance in methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from 18 hospitals in Gdansk area was determined. The study was carried out on 190 MRSA isolated in 1997-2000 from various clinical samples. The strains were tested for high-level mupirocin resistance by 200 micrograms mupirocin disc. The minimum inhibitory concentrations (MIC) for methicillin were estimated by agar dilution. Sensitivity to other antibiotics was determined in disc-diffusion method and to vancomycin in agar dilution method additionally. The strains were typed by set of 10 experimental phages and compared by the method of PCR-RFLP analysis of coagulase gen restriction fragment length polymorphism. There were low frequency of high-level mupirocin resistance in MRSA strains (4.7%) that were found only in 3 hospitals, in 6 patients. All of them were high-resistant also to methicillin and resistant to doxycyclin, gentamycin, erytromycin, klindamycin, ciprofloksacin, rifampicin, resistant or intermediate sensitive to fusidic acid but sensitive to vancomycin, teikoplanin and bacitracin. The origin all of the MRSA strains high-resistant to mupirocin probably was the same, except one strain, because they were belonged to one genetic type and possessed the same phage pattern.