[Characteristics of clumping factor- and protein A-negative methicillin-resistant Staphylococcus aureus identified in Poland]. / Caractéristiques des souches de Staphylococcus aureus facteur agglutinant- et protéine- A-négatifs résistants à la méthicilline isolées en Pologne.

AIM: The aim of this study was to investigate the phenotypic and genotypic properties of clumping factor- and protein A- negative MRSA. METHODS: Two hundred ninety-four strains were studied. The production of protein A was determined by immunoblotting, clumping factor (CF) and coagulase by conventional tests with rabbit plasma. The presence of nuc gene and mecA gene was identified by PCR. The presence of clfA and spa genes was determined for CF- and protein A-negative strains and then phage typing was performed using two sets of phages and gene typing by RFLP of the spa gene. RESULTS: Sixteen (5.4%) CF- and protein A-negative MRSA were isolated. All of them produced coagulase and thermostable nuclease (except for one). nuc, mecA, spa and clfA genes were observed in all strains. The same RLFP pattern, but with different phage types and resistance profiles to antibiotics, was observed in all strains. CONCLUSIONS: CF- and protein A-negative methicillin resistant S. aureus isolated in Poland do not come from a single clone and the absence of CF and protein A is not caused by the deletion of clfA and spa genes.

[Drug resistance of group B streptococci]. / Lekoopornosc paciorkowców grupy B.

The group B streptococci (GBS) are one of the main infections' factors in neonates. The source of the infection is mother and the infection follows during childbirth. In many countries various methods of prophylaxis in reference to mother and neonate are introduced. Thus far the widest application have found a method of giving to pregnant women penicillins or cephalosporins. The purpose of the present work was to study how the drug-resistant of the GBS forms in our country and if using antibiotics in prophylactic purposes without doing any antibiograms is burdened with risk, resulted from the possibility of meeting resistant strains. 365 strains of the GBS, derived from colonised neonates, pregnant women and clinical materials were put to the scientific research. 120 animal strains isolated from events of mastitis in cows, represented the second group. The GBS, derived from pregnant women were sensitive to every examined antibiotics except lincomycin (5.9%). Amid the streptococcus isolated from neonates there were some strains resistant to every examined antibiotic (1.9-4.4%). More often resistant strains were isolated from clinical materials (3.9-9.0%). The highest percentage of the resistant strains was identified in animals (7.5-20.0%). The GBS as a rule were insensitive to cotrimoxasolum. As follows from our scientific researches, the GBS derived from people as a rule are sensitive to used antibiotics. But there were some strains resistant even to several antibiotics. Possibility of meeting these strains should be taken into consideration as in prophylaxis as in treatment of the infections. Amid the animal strains of the GBS the phenomenon of the resistance to antibiotics is more often met than in human ones.

[Biochemical typing as a method of differentiating group B streptococcus]. / Typowanie biochemiczne jako metoda róznicowania paciorkowców grupy B.

In epidemiological studies on the group B streptococcus the serological typing is used. The paper present the results of a study on usefulness of biochemical typing for differentiation of the group B streptococcus. For that purpose, 210 strains descended from colonized infants and pregnant women were put to typing with both of mentioned methods. We showed that each of the method distinguishes similar number of biotypes and serotypes. However, ought to be marked that significant number of strains (93.8%) belonged to the three out of eight biochemical types. Similar results were achieved in serological typing, three of the most numerous serotypes contained 81.4% strains. Analysis of the relationship between serological and biochemical types did not reveal statistical association because the strains belonged to various serotypes. Obtained results show that both methods of typing--biochemical and serological, have similar value in differentiation of the strains. The method of biochemical typing is quite simple and can be used in laboratory conditions.

[Predominance of multidrug resistant strains with reduced susceptibility to fusidic acid among methicillin-resistant Staphylococcus aureus strains (MRSA) isolated in the Gdánsk region]. / Dominacja wieloopornych szczepów o zmniejszonej wrazliwosci na kwas fusydowy wsród metycylinoopornych gronkowców zlocistych (MRSA) izolowanych w regionie gdanskim.

The aim of the present study was to follow the changes in the drug resistance among the methicillin-resistant Staphylococcus aureus strains (MRSA) isolated from clinical samples in various hospitals during 8 years, with particular consideration of Gdansk area. The study was carried out on 225 strains of MRSA from which 95 were isolated in the years 1990-1995 and 130 in the years 1997-1998. The drug susceptibility was determined by the disc-diffusion method. The sensitivity to fusidic acid was determined by both disc-diffusion method and minimal inhibitory concentration (MIC) using agar dilutions. The results obtained show that in 1997-1998 in the hospitals of Gdansk area have been appeared MRSA strains which have not occurred before. These strains were intermediately sensitive or resistant to fusidic acid and simultaneously resistant to doxycycline, gentamicin, erythromycin, clindamycin, ciprofloxacin and rifampin. They represented 66.2% of all MRSA strains isolated in 1997-1998 and were present in majority of the hospitals monitored. Only in this group of staphylococci the strains additionally resistant to mupirocin (6.2%) occurred. Among the MRSA strains with reduced susceptibility to fusidic acid 64.4% were intermediately sensitive (MIC 8-16 mg/L) and 15.4% were resistant to this drug (MIC > 16). In 1997-1998 the percentages of MRSA strains resistant to rifampin, clindamycin and ciprofloxacin increased significantly from 12.6% to 90.8%, from 42.1% to 92.3% and from 18.9% to 92.3% respectively. The percentage of the chloramphenicol resistant strains decreased from 14.7% to 0.8%. Like in 1990-1995, the MRSA strains resistant to vancomycin and teicoplanin were not found out in the same period of time.

[Changes of methicillin-resistant Staphylococcus aureus (MRSA) phagotypes in Gdansk in 1990-1998]. / Zmiany fagotypów gronkowców zlocistych, opornych na metycyline (MRSA) w regionie Gdanskim w latach 1990-1998.

The aim of the study was the assessment of changes in the occurrence of various MRSA phagotypes in hospitals in the Gdansk area in 1990-1998. The study was carried out on 175 MRSA strains: 45 strains isolated in 1990-1995 and 130 in 1997-1998. The studied staphylococci were obtained from various clinical materials from patients in 18 hospitals. Phagotyping was done with a set of 10 experimental phages from MRSA strains obtained from the Central Public Health Laboratory in London. Drug-resistance was determined by the disc-diffusion method and in case of strains with medium susceptibility to fusidic acid the minimal inhibitory concentration (MIC) was determined by serial dilutions on solid medium. The study showed among MRSA strains isolated in 1997-1998 a new, previously not known strains with phage pattern MR25/M5 predominated (57.7%). Its presence was found in various hospitals in that area. MRSA belonging to MR25/M5 phagotype were mostly resistant to doxycycline, gentamycin, erythromycin, clindamycin, ciprofloxacin, rifampicin and were resistant or only had medium susceptibility to fusidic acid. The MIC values for strains with medium susceptibility to fusidic acid (4-8 micrograms/ml) showed an evident decrease of the susceptibility to this antibiotic, formerly common in MRSA. At the same time, in 1997-1998 a considerable decrease was observed of the number of MRSA strains belonging to MR8/MR12/MR25/30/33/38/M5/622 phagotype (from 31.1% to 0.8%), and disappearance of strains with phagotypes MR25/56B/M3 and MR8/MR25/622, which in 1990-1995 accounted for 15.6% of the studied staphylococci.

[Predominance of phage group II in Staphylococcus strains isolated from humans]. / Dominacja II grupy fagowej wsród szczepów Staphylococcus aureus izolowanych od ludzi.

2545 strains of Staphylococcus aureus isolated from 24 Polish laboratories in the years 1994-95 were investigated. Phage typing was performed according to the method of Blair and Williams using the present basic set of typing phages and additional phages 88, 89 and 187. The phages were employed in concentrations of RTD and 100 x RTD. The predominance of phage group II, reported elsewhere since 1980-ties, was found in the present study (23.6%). Strains of group III were second in frequency (16.6%), whereas strains of groups I and V, as well as type 95 occurred in small percentage (7.6%, 4% and 3.4% respectively). Strains of groups II and V have been rarely lysed by phages belonging to other groups. The use of additional phages resulted in typability of strains by 7.9%. Percentage of non-typable strains was high and amounted to 22.0%.

[Evaluation of the usefulness of new international experimental phages for typing methicillin resistant Staphylococcus aureus (MRSA)]. / Ocena przydatnosci nowego, miedzynarodowego zestawu eksperymentalnych fagów do typowania opornych na metycyline gronkowców zlocistych (MRSA).

The aim of the study was to determine the usefulness of the set of experimental phages obtained from the Central Public Health Laboratory in London for typing of MRSA strains in Poland. The study was performed on 150 MRSA strains isolated from various clinical materials in various regions of the country. The set of 10 experimental phages and the international basic set of 23 phages were used for typing. The results of the study showed that 76.8% of MRSA strains were typing with the experimental set of phages. The frequency of inhibition reactions was 19.9%. Only 3.3% of the strains were nontypable with the new phages while nearly half of the studied strains were nontypable with the basic set of phages. The studied strains were divided into 19 phagotypes. There was a high frequency of typable strains among MRSA typable and nontypable strains and those inhibited by the basic set of phages (71.4%-85.7%). These data indicate that the set of 10 experimental phages is useful for typing of MRSA strains isolated in Poland except for phage M3 which failed to react with almost all the strains and should be excluded from the proposed set.

[Use of Polymerase chain reaction (PCR) for detection of Mycobacterium tuberculosis complex from patient materials]. / Zastosowanie lancuchowej reakcji polimerazy (PCR) do wykrywania Mycobacterium tuberculosis complex w materialach od chorych.

The polymerase chain reaction (PCR) and direct culture were applied for detection of Mycobacterium tuberculosis complex in samples obtained from patients with suspicion of pulmonary and extra-pulmonary tuberculosis. In the reaction of amplification IS6110 was applied as a target region, and PCR reaction products were of the size of 123 bp and 317 bp. A total of 278 samples (158 sputum, 36 urine, 25 pleural effusion, 23 bronchial washings, 8 blood, 4 stomach washings, 3 cerebrospinal fluid, 2 fragments of skin, 2 pleural effusion, and 17 others samples) from 181 patients were tested. Mycobacterium tuberculosis complex was detected by PCR in 144 out of 278 samples and by culture in 68 of 278 samples. The PCR test enabled a rapid and sensitive diagnosis particularly in a number of samples which were negative on culture.

[Asymptomatic bacteriuria in women diagnosed with systemic lupus erythematosus (SLE)]. / Bakteriomocz bezobjawowy u kobiet z rozpoznaniem tocznia rumieniowatego ukladowego (T.R.U.).

The aims of our research were: estimation of asymptomatic bacteriuria (a.b.) incidence in population of women with systemic lupus erythematodes (SLE), evaluation its clinical significance and examination of bacterial colonisation of nostrils and pharynx in SLE patients with a.b. 85 women aged 24-77 (mean 49.3) with mean SLE duration 7.8 (range 1-32) years were examined. All of them fulfilled ARA criteria for the classification of SLE. Among group of patients with a.b. were counted women who had significant bacteriuria > or = 10(4) in ml urine in two cultures. Asymptomatic bacteriuria was found in 14 cases of 85 women with SLE (16.5%). In two following urine cultures bacteria from family Enterobacteriaceae were dominated: the same types of bacteria were in 85.7%--bacteriuria persistens, in others 14.3% were observed change of bacteria--bacteriuria transistens. In 9 from 14 patients with (64.3%) a.b. very massive growth of Staphylococcus aureus in culture from vestibulae of the nose swab was, in other cultures very massive growth of physiological flora was seen. All patients with a.b. were in clinical remission of SLE and they had no clinical symptoms of infection in urinary tract in 5 months of observation. However clinical significance of asymptomatic bacteriuria and pathogenic bacteria colonisation of nostrils as a precedence to symptomatic infections needs further investigations.

[Deamination of adenine and adenosine in staphylococci]. / Deaminacja adeniny i adenozyny u gronkowców.

Deaminations of adenine and adenosine by pattern strains of 24 staphylococcal species, were tested. During 3 hours of incubation of the suspensions of 8 staphylococci with adenine the liberation of ammonia occurred. The same staphylococci accumulated ammonia in the incubation medium with adenosine. The Staphylococcus intermedius PCM 2405 strain as opposite to the Staphylococcus aureus 536 strain in the media with adenine or adenosine accumulated hypoxanthine or inosine, respectively and ammonia. These results indicated that adenine deaminase (adenase) and adenosine deaminase activities were associated with the cells of the Staphylococcus intermedius PCM 2405 strain. Staphylococci were heterogeneous within three species groups with respect to adenine and adenosine deaminations. Adenine and adenosine determinations were absent in staphylococci belonging to the Staphylococcus simulans species group.

[Sensitivity to disinfectants of methicillin resistant Staphylococcus aureus strains (MRSA) and gentamicin resistant strains]. / Wrazliwosc na srodki dezynfekcyjne metycylinoopornych gronkowców zlocistych (MRSA) i szczepów opornych na gentamycyne.

The aim of the study was to find out whether methicillin-resistant S. aureus strains (MRSA) are tolerant in a higher degree to disinfectants, and whether a correlation exists between lower sensitivity to these agents and resistance to gentamicin. The study was carried out on 30 strains of MRSA and 20 of MSSA isolated from various clinical materials in various regions of the country. Among the studied MRSA 24 strains were resistant and 6 were sensitive to gentamicin, and in MSSA 3 strains were resistant and 17 sensitive to this antibiotic. The sensitivity to four disinfectants: Manusan, Sterinol, Septyl R and Lysoformin Spezial was determined by measurement of MIC (minimal inhibitory concentration) in agar medium. Most MRSA in Poland showed decreased sensitivity to these disinfectants. Among gentamicin-sensitive and resistant MRSA strains the proportions of strains with higher tolerance of three disinfectants (Manusan, Sterinol and Lysoformin Spezial) were very similar. Reduced sensitivity to disinfectants was found in all gentamicin-resistant MSSA. These data indicate that S. aureus strains possess various mechanisms of tolerance of disinfectants. Nearly half the studied strains (46%) had decreased sensitivity to all three preparations (Manusan, Sterinol and Lysoformin Spezial) belonging to various chemical groups this seems to indicate that increased tolerance to these disinfectants is a non-specific feature of S. aureus strains.

[Bacterial diagnosis of tuberculosis using the PCR technique]. / Diagnostyka bakteriologiczna gruzlicy technika PCR.

At the beginning of 80-ties new system BACTEC for mycobacteria culture has been introduced. So time of this culture has been shortened to 1-3 weeks. At the end 80-ties Polymerase Chain Reaction (PCR) technique was started and at present it is used in many laboratories in order to get information about genetic material of mycobacteria in different biological samples. Most often insertive factor IS 6110, located in chromosome Myc. tub. complex, is used. DNA amplification occurs in three stages: 1) DNA denaturation. 2) addition of starters (primers): 3) elongation of starter with d NTP synthesis. PCR technique identifies genetic material (DNA) of mycobacterium which is presented in a given biological sample. Positive PCR result not necessarily means that active disease takes place.

Rapid assay for detection of methicillin-resistant Staphylococcus aureus using multiplex PCR.

The presence or absence of the mecA gene, the determinant of resistance to all beta-lactam antibiotics, was examined in clinical isolates of Staphylococcus aureus by multiplex polymerase chain reaction (MPCR). Two pairs of primers were used, which yielded two specific products; a 280-bp nuc- based PCR fragment (amplification product of the nuc gene encoding specific Staphylococcus aureus nuclease) and a 533-bp mecA-based PCR fragment (amplification product of the mecA gene). The MPCR system was designed to be incorporated into the work flow in clinical diagnostic laboratories as a routine analysis.

[Occurrence of rota-I adenoviruses in diarrhea states of children]. / Wystepowanie rota- I adenowirusów w biegunkowych stanach u dzieci.

98 children aged 6 weeks to 5 years with diarrhoea were examined. Rotaviruses and adenoviruses were detected by latex tests Rotalex and Adenolex produced by Orion Diagnostica. Rotaviruses were found in 20.4% of cases, most frequently in children of the age from 6 to 18 months. Adenoviruses were found in 11.2% of cases, most frequently in children of the age from 18 months to 5 years, mainly in mixed infection by- and adenoviruses. Viruses infection were most rare in infants aged from 6 weeks to 6 months. In 4.1% studied cases the coexisting of viral and bacterial infection was observed (K. pneumoniae, E. coli).

[Colonization with group B streptococci in neonates and premature infants from selected neonatal departments]. / Kolonizacja paciorkowcami grupy B noworodków i wczesniaków w wybranych oddzialach noworodków.

Group B streptococci are considered an important etiological agent of sepsis and meningitis in neonates and, particularly, in premature infants. There is a close correlation between colonization with these bacteria and the frequency of symptomatic infection. It is estimated that symptomatic infections occur in 1.0% of colonised neonates. The purpose of this work was to investigate the frequency of neonate colonization with group B streptococci for determination of the risk of symptomatic infection.

The clinical course of untreated asymptomatic bacteriuria in diabetic patients--14-year follow-up.

The present study aimed at accessing the natural history of untreated asymptomatic bacteriuria in diabetic patients observed for 14 years. Two groups of diabetic patients were examined. Group I comprised 53 patients with significant bacteriuria, without clinical symptoms of urinary tract infection. Group II consisted of 54 patients with sterile urine. All patients were clinically and bacteriologically examined every 3-6 months for 14 years. The results obtained in this study have proved that clinical symptoms of acute pyelonephritis occur with similar frequency in both groups (group I-6, group II-5 cases). No deterioration of kidney function was found. Arterial hypertension did not differ significantly in both groups at the beginning and end of the follow-up. The usefulness of antibacterial treatment of asymptomatic bacteriuria in diabetic patients is not adequately documented.

[Susceptibility of bacteria isolated from clinical material to ciprofloxacin]. / Wrazliwosc bakterii izolowanych z materialu klinicznego na cyprofloksacyne.

The aim of the study was to evaluate susceptibility to ciprofloxacin of bacteria species isolated from different specimens of clinical materials. The investigated strains (830) were identified using conventional methods. The antibacterial activity of ciprofloxacin was studied by the method of drug dilution in Mueller-Hinton agar. Among aerobic bacteria, only 5 strains (0.6%) were resistant to ciprofloxacin in concentration 4 micrograms/ml; two belonged to E. coli species, two to S. aureus and one was a Streptococcus group B., Corynebacterium sp. MIC90 = 0.12 microgram/ml turned out to be most resistant, next, Gram-negative rods MIC90 = 0.5 microgram/ml, coagulase-negative and positive staphylococci and Neisseria sp. MIC90 = 1.0 microgram/ml. Enterococcus faecalis and anaerobes were the least susceptible to ciprofloxacin. Their MIC90 was 2 and 16 micrograms/ml respectively. Among 20 anaerobic strains, up to 10 were resistant to ciprofloxacin, mainly from Bacteroides and Clostridium genus.

[Occurrence of methicillin-resistant strains in Poland and their characteristics]. / Wystepowanie metycylinoopornych gronkowców w Polsce i ich charakterystyka.

The investigation was performed on 923 strains of S. aureus isolated from clinical material obtained from several regions of Poland. Resistance to methicillin was tested by a dilution method on a solid Mueller-Hinton medium supplemented with 2% of NaCl. Strains exhibiting MIC higher than 4 micrograms/ml were determined as resistant. Resistance to other antibiotics (P, Am, CB, CF, CH, MA, Ge, Bs, E, L, Dx) was tested by a disc method. Bacteriophage typing of S. aureus was performed by a method described by Blair and Williams in RTD and RTD x 100, using basic set of phages and additional phages (88, 89, 187). MRSA were present in various regions of the country with similar frequency (from 3.3% to 8.3%). In one center only the percentage was as high as 52.3%. High percentage of MRSA was noted in burned patients (59.9) and these strains were obtained at one center. Within the MRSA III phage group was dominating as well as non-typable strains and inhibited at 100 x RTD-35.4%. MRSA most frequently were typing with phages 88-40.9%, 89-35.0%, 85-24.0%, and they rarely belonged to the phage group II. Among MRSA strains higher percentage of antibiotic-resistance was noted, as compared with other strains. About 60% of MRSA strains were resistant to 6-8 antibiotics. The dominating resistance concerned penicillins, cephalosporins, aminoglycosides, tetracyclines and erythromycin.

[Bacteriophage types of staphylococcus aureus in neonatal wards of hospitals in Gdansk region during the years 1984-1991]. / Typy bakteriofagowe gronkowców zlocistych w oddzialach noworodkowych szpitali województwa gdanskiego w latach 1984-1991.

Investigated 3558 strains of S. aureus were obtained from healthy newborn infants in neonatal wards. The staphylococci were classified as S. aureus on the basis of a positive coagulase test. Bacteriophage typing was performed with a basic set of phages and an additional phage (187), according to the method Blair and Williams, in RTD and RTD x 100. Additionally, susceptibility of staphylococcal phagotypes isolated in the years 1988-1991 to antibiotic from penicillin and cephalosporin groups, trimethoprim-sulphamethoxazole, erythromycin, lincomycin and doxycycline, was determined by disc diffusion method. Among tested strains phagotypes of group II dominated. They constituted 31.3% of all present phagotypes. Second in frequency were phagotypes of group V and III (13.2% and 10.0%). Large number of staphylococci (22.9%) was typable at the same time with phages belonging to different phage groups. Tested staphylococci were highly resistant to penicillin (98.8%), ampicillin (98.9%), cloxacillin (38.6%), carbenicillin (32.6%) and doxycycline (45.6%).

[Susceptibility of methicillin-sensitive and methicillin-resistant staphylococci to ciprofloxacin]. / Wrazliwosc gronkowców metycylino-wrazliwych i metycylino-opornych na ciprofloksacyne.

Investigations were carried on 352 strains of Staphylococcus aureus and 105 strains of coagulase-negative staphylococci, isolated from various clinical materials and derived from some regions of the country. Methicillin-resistance was tested by an antibiotic dilution method in solid Mueller-Hinton medium with addition of 2% NaCl. Staphylococci with MIC higher than 4 micrograms/ml were considered as resistant. The same method for testing resistance to ciprofloxacin was used. Only one strain (S. aureus) was resistant to both ciprofloxacin and methicillin. All remaining strains of staphylococci were ciprofloxacin-susceptible. The MIC for all of them was not higher than 2 micrograms/ml, regardless of resistance to methicillin. Some slight differences in MIC50 and MIC90 values were found between MRSA and MSSA and they were, respectively, 1 microgram/ml and 0.25 microgram/ml, and 2 micrograms/ml and 1 microgram/ml. Mean MIC of ciprofloxacin for MRSA was 1.1 microgram/ml and for MSSA it amounted to 0.4 microgram/ml. Range of MIC was following: MRSA--0.12 microgram/ml--16 micrograms/ml and MSSA--0.12-2 micrograms/ml. Coagulase-negative staphylococci, both methicillin-resistant and methicillin-sensitive, exhibited same value for MIC50, MIC90 and MIC range and they were following: 0.5 microgram/ml, 1 microgram/ml and 0.12-1 micrograms/ml. Both groups differed slightly in mean MIC values which was 0.6 microgram/ml for methicillin-resistant strains and 0.47 microgram/ml for methicillin-sensitive staphylococci. It seems that within staphylococci isolated in Poland there is no correlation between resistance to methicillin and ciprofloxacin, which is frequently pointed out by other authors.