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A Tomasz

Publications and source records attributed to A Tomasz.

At least 73 records · Page 4Linked to original sources
PubMed1998

Suppression of glycopeptide resistance in a highly teicoplanin-resistant mutant of Staphylococcus aureus by transposon inactivation of genes involved in cell wall synthesis.

The teicoplanin-resistant laboratory mutant TNM of Staphylococcus aureus strain COL (minimal inhibitory concentration for teicoplanin increased from 3 to 200 microg/ml) produced an abnormal peptidoglycan in which the proportion of cross-linked oligomeric muropeptides (pentameric and higher than pentameric species), representing approximately 60% of all muropeptide species in the parental strain, was reduced to approximately 17% in the mutant. In parallel, there was an increase in the representation of the monomeric muropeptides from 4% (in the parent) to 20% in the resistant strain. The mutant cell wall showed greatly increased porosity for the detergent extraction of cytoplasmic proteins, and this property was abolished in a Tn551 insertional derivative of TNM, which was selected for reduced (parental level) teicoplanin resistance. Transposon inactivation of the global regulatory genes Sigma-B and sar, and several genes involved in early steps of staphylococcal peptidoglycan synthesis, all caused extensive reduction of teicoplanin resistance in mutant TNM, in some cases to levels close to or below the MIC value of the parental strain.

Anti-Bacterial Agents↗
PubMed1998

Molecular epidemiologic characterization of penicillin-resistant Streptococcus pneumoniae invasive pediatric isolates recovered in six Latin-American countries: an overview. PAHO/Rockefeller University Workshop. Pan American Health Organization.

The Pan American Health Organization (PAHO) has conducted a study of Streptococcus pneumoniae in six Latin-American countries: Argentina, Brazil, Chile, Colombia, Mexico, and Uruguay. Sterile site isolates from children aged < or =5 years showing clinical symptoms of pneumonia (as defined by the clinical criteria of WHO), meningitis, sepsis or bacteremia (without infectious foci), arthritis, and peritonitis were the source of most of the invasive pneumococcal isolates collected between the end of 1993 and 1996 in the six participating countries. Partial characterization of these isolates (antibiotic resistance and serotyping) have already been described (Microbial Drug Resistance 3:(2):131-163, 1997). In the next phase of the study, 326 S. pneumoniae isolates with reduced penicillin susceptibility were transferred to the Laboratory of Microbiology at The Rockefeller University for molecular characterization, and a summary and overview of the findings is described in this article. Some of the most interesting findings were as follows: (1) There was a surprisingly high representation of two internationally spread clones, which made up >80% of the strains with penicillin MIC of 1 microg/ml or higher; most of these isolates were recovered in large cities, supporting the likelihood that the source of these clones is through international travel. (2) The frequency of resistance to trimethoprim/sulfamethoxazole was extremely high (present in 85% of all isolates with decreased penicillin susceptibility). (3) None of these isolates was resistant to ofloxacin, and macrolide resistance was rare (present in 6.4% of the isolates). (4) There was an apparent inverse relationship between level of penicillin resistance and genetic diversity. (5) There were striking differences in the "microbiologic profiles" of the six different Latin-American countries.

Child↗
PubMed1998

Geographic distribution of penicillin resistance of Streptococcus pneumoniae in Brazil: genetic relatedness.

From January 1993 through December 1996, 1,252 Streptococcus pneumoniae strains from different geographic regions of Brazil were studied for penicillin (Pen) susceptibility. All pneumococci were isolated from normally sterile fluids from patients, newborns to 88 years old. Pen resistance (R) had a mean rate of 15.1%, with 14.5% of strains showing intermediate level Pen-R and 0.6% showing high-level Pen-R. Similar Pen-R rates were observed in different regions of the country, in the range of 9.5% to 17.1%. A Pen-R increase was noted from 9.6% in 1993 to 20.6% in 1996. Pen-R was mostly associated to serotypes 6B, 14, 19A, and 23F (89%). Chromosomal DNA relatedness of Pen-R strains was determined by pulsed field gel electrophoresis (PFGE). High genetic diversity was identified, being represented by 27 patterns among the 92 strains. Two important features were observed: the predominance of relatively low-level Pen MIC (range 0.1-0.5 mg/L) in 86 of the 92 strains, and the presence of 60.8% as four major PFGE clusters unique to Brazil. Another feature was the geographic spread of these clusters over large distances in the country. The city of São Paulo seems to be a Pen-R focus (18.4%) in Brazil. Only two strains representing the international clone B widely spread in France, Portugal, and Spain, belonging to serotype 14, were found.

Brazil↗
PubMed1998

The apparent importation of penicillin-resistant capsular type 14 Spanish/French clone of Streptococcus pneumoniae into Uruguay in the early 1990s.

Since 1987, Streptococcus pneumoniae (SPN) has been monitored in pediatric and adult populations. In 1994, the SIREVA/PAHO surveillance allowed the assessment of a dramatic increase of penicillin resistance, mainly associated to SPN type 14 also resistance to trimethoprim/sulfamethoxazole. To determine the genetic relatedness of the resistant isolates, analysis of SmaI digests of 61 isolates was performed by pulsed field gel electrophoresis (PFGE). Twelve different profiles were identified. The type 14 resistant SPN (n = 47) displayed a common B pattern, with 15 genetically related subtypes (1-6 bands of difference), with B1 the predominant one (51%). One of the subtypes (B12) was indistinguishable from a French isolate (M13P). This strain represented a South European clone, identified in the late 1980s, that also included serogroup 9 isolates. The type 23F isolates (n = 3) were identical with a representative strain of the well-characterized intercontinental type 23F clone. Neither the type 14 penicillin-susceptible (n = 4) nor the resistant SPN of other serotypes (n = 10) belonged to the B pattern. In summary, the results suggest that the dramatic increase of penicillin resistance in Uruguay depends on the introduction and spread of a type 14 clone, apparently imported from the south of Europe. Follow-up of this phenomenon is mandatory from the point of view of epidemiology and is also a priority for biologic research.

Electrophoresis, Gel, Pulsed-Field↗
PubMed1998

Penicillin-resistant Streptococcus pneumoniae in Argentina: frequent occurrence of an internationally spread serotype 14 clone.

Six Latin-American countries participated in an epidemiological surveillance study conducted by the Pan American Health Organization in order to determine the relative prevalence of capsular types and antimicrobial resistance patterns of Streptococcus pneumoniae (SPN) causing invasive infections in children <5 years of age. In Argentina, the incidence of penicillin resistance (PR) was 24.4%, and it was significantly associated with serotype 14 (p < 0.001). The chromosomal DNA of 56 of those SPN isolates, 39 PR and 17 susceptible, was digested with SmaI and resolved by PFGE. Eighty-two percent (32/39) of the PR isolates shared characteristics with the widely spread International Spanish/French clone (clone B). All members of clone B except one expressed serotype 14, with the exception of one isolate that expressed serotype 19F and probably resulted from an in vivo capsular transformation event. Only a single isolate shared features with the 23F International Spanish/USA clone (clone A). The 17 penicillin-susceptible (PS) SPN isolates presented an enormous degree of variation in the chromosomal background, expressing 12 serotypes and 13 PFGE patterns. The data suggest that over 80% of the SPN-PR isolates in Argentina were imported, and this confirms the importance of the geographic spread of SPN clones in South America.

Argentina↗
PubMed1998

Penicillin-resistant Streptococcus pneumoniae in Colombia: presence of international epidemic clones. Colombian pneumococcal study group.

The global spread of multidrug-resistant Streptococcus pneumoniae clones is well documented in the literature. A study to determine type distribution and antimicrobial susceptibility of invasive pneumococcal isolates from Colombian children under the age of 5 was conducted from 1994 to 1996. Health centers in Santa Fe de Bogota, Medellin, Cali, and other cities collected 409 Streptococcus pneumoniae isolates. Diminished susceptibility to penicillin (DSP) was 15.6%; from these, 11.5% showed intermediate-level resistance (ILR) and 4.1% showed high-level resistance (HLR). Fifty-nine of the DSP isolates were examined by pulse field gel electrophoresis (PFGE). Capsular isolate types were 23F (54%), 14 (24%), 19F (10%), 6B (7%), 9V (3%), and 34 (2%). PFGE analysis revealed that 8 isolates shared the Spanish/USA international clone's characteristic features: PFGE pattern type A, serotype 23F; 87.5% exhibited HLR for penicillin, and all were resistant to trimethoprim/sulfamethoxazole (TMP-SMX), tetracycline, and chloramphenicol. Another 7 isolates showed the French/Spanish international clone's features: PFGE pattern type B, 2 of them being serotype 9V; and 5 type 14; HLR to penicillin was 71%, and all proved resistant to TMP-SMX. A large cluster of 24 isolates (41% of all isolates examined) shared a common PFGE type C, with 14 subtypes; all but one, serotype 34, were serotype 23F and had ILR to penicillin; 58% were resistant to TMP-SMX and 50% to tetracycline, but none presented erythromycin or chloramphenicol resistance. The remaining 20 isolates could be grouped into 12 different PFGE types; ILR was shown in 75% of isolates, 70% were resistant to TMP-SMX and to tetracycline, 15% were resistant to erythromycin, and none were resistant to chloramphenicol. These data suggest that some Colombian isolates are clonally related to two of the well-known international epidemic S. pneumoniae clones.

Colombia↗
PubMed1998

Predominance of the multiresistant 23F international clone of Streptococcus pneumoniae among isolates from Mexico.

During a surveillance study to determine the relative prevalence of capsular types of Streptococcus pneumoniae and antimicrobial susceptibility of invasive isolates in children <5 years old in Mexico City, 220 isolates were collected. The serotype 23F was the most common found, followed by types 6A + B, 14, 19F, and 19A. Diminished susceptibility to penicillin was detected in 106 isolates (48.2%), and high penicillin resistance was found in 49 strains (22.2%), 31 belonging to type 23F. Resistance was also observed to erythromycin (13.1%), to chloramphenicol (43.1%), and to cefotaxime (10.9%). No strains were resistant to ofloxacin or vancomycin. Forty-four of the highly penicillin resistant isolates (penicillin MIC > or =2.0 microg/ml) were examined with molecular fingerprinting techniques; 29 (65.9%) of these isolates (all except two strains) were serotype 23F and shared subtype variants of PFGE type A characteristic of the internationally spread Spanish/USA clone of S. pneumoniae. These strains were also resistant to trimethoprim/sulfametoxasole (TMP/SMX), chloramphenicol, and tetracycline, and most of them were susceptible to erythromycin. Another 6 of the highly penicillin-resistant strains (serogroups 9 and 14) showed PFGE fingerprints and antimicrobial susceptibility pattern characteristic of a second internationally spread clone (French/Spanish clone) and carried resistance to penicillin and TMP/SMX. The rest of the 9 penicillin-resistant isolates were represented by 7 distinct additional PFGE types. The findings suggest that almost 80% of all highly penicillin resistant strains may have been "imported" into Mexico.

Child, Preschool↗
PubMed1998

Molecular characterization of penicillin-resistant Streptococcus pneumoniae isolates causing respiratory disease in the United States.

Three hundred twenty-eight (328) penicillin-resistant Streptococcus pneumoniae isolates collected in 39 states of the United States between October, 1996, and March, 1997, from (mostly adult) patients with respiratory disease were characterized by microbiological, serological, and molecular fingerprinting techniques, including determination of chromosomal macrorestriction pattern with pulsed-field gel electrophoresis (PFGE) and hybridization with DNA probes specific for various antibiotic resistance genes. The overwhelming majority of the isolates were in five serogroups (23, 6, 19, 9, 14). All isolates had penicillin MIC values of at least 2 microg/ml, but the collection also included isolates with MIC values as high as 16 microg/ml. Virtually all isolates (96.6%) were resistant to trimethoprim/sulfamethoxazole (SXT) and many isolates were also resistant to chloramphenicol (43%), tetracycline (55%), and erythromycin (65%). Resistance to levofloxacin was extremely rare. The molecular fingerprinting methods showed that a surprisingly large proportion (167 out of 328, or 50.9%) of the isolates belonged to two international epidemic clones of S. pneumoniae: clone A (127, or 38.7%) with properties indistinguishable from that of the 23F multiresistant "Spanish/USA" clone widely spread in Europe, Asia, Latin America, and South Africa, and clone B (40, or 12.2%) belonging to the "French" serogroup 9/14 clone widely spread in Europe and South America. Virtually all members of clone A were also resistant to chloramphenicol (cat+), tetracycline (tetM+), and SXT, and about 75% were also resistant to erythromycin (mefE+ or ermB+). Close to 30% (39 out of 127) of the clone A isolates expressed anomalous serotypes (primarily serotypes 19 and 14, and nontypable) and most likely represented spontaneous capsular transformants. Most of the 40 isolates (35/40) belonging to clone B expressed serotype 9, with five of the isolates expressing serotypes 14 or 19, or were nontypable. All members of this clone were resistant to penicillin and SXT with only occasional isolates showing resistance to macrolides, tetracycline, and chloramphenicol. The combination of microbiological tests and DNA hybridizations also allowed the identification of unusual strains, for instance, isolates that reacted with the tetM or mefE DNA probes without showing phenotypic antibiotic resistance, an isolate showing phenotypic macrolide resistance without hybridizing with either the ermB or mefE DNA probes, or isolates that hybridized with both of these DNA probes. In addition to clones A and B, another large portion of the S. pneumoniae isolates (112 of 328, or 34.1%) was represented by eight clusters, each with a unique PFGE type. These clusters, together with the clone A and clone B isolates, made up 85% of all the penicillin-resistant isolates identified in this survey in the United States. Both international clones and the unique clusters showed wide geographic dispersal: Clone A was present in 30 of the 39 states and clone B in 18. The data suggest that the major mode of spread of penicillin-resistant pneumococci in the United States is by clonal expansion and that the most significant components (clones A and B) have been imported into the United States from abroad.

Adult↗
PubMed1998

Antibacterial efficacy of nisin against multidrug-resistant Gram-positive pathogens.

The rapid spread of multidrug-resistant bacterial pathogens necessitates the search for alternative antibacterial agents. We examined the efficacy of the antibiotic nisin against 56 multidrug-resistant isolates of Streptococcus pneumoniae, 33 Staphylococcus aureus and 29 vancomycin-resistant Enterococcus faecium and Enterococcus faecalis isolates. The test strains represented a large variety of clonal types (as determined by a combination of DNA fingerprints) isolated from a variety of geographic sources, and included some of the major internationally-spread multiresistant epidemic clones of S. pneumoniae and methicillin-resistant S. aureus (MRSA), MRSA strains resistant to over 16 generically distinct antibacterial agents, and enterococcal strains resistant to all currently available chemotherapeutic agents including glycopeptides. In the overwhelming majority of cases, treatment of growing cultures with nisin at 1 mg/L (S. pneumoniae) or 10-20 mg/L (in MRSA and enterococci) caused extensive (10(3)- to 10(4)-fold) loss of viable titre accompanied by various degrees of loss in the optical density of the cultures, which was most extensive in pneumococci (>90%) and least extensive (40-50%) in enterococci. Nevertheless, extensive variation in rates of nisin-induced autolysis was observed in each bacterial species. Serial exposure of a penicillin-susceptible strain of S. pneumoniae to nisin (1 mg/L) in liquid culture resulted in the rapid appearance of stable nisin-resistant mutants in which the MIC increased from 0.4 to 6.4 mg/L and the resistance trait was transferable by genetic transformation.

Anti-Bacterial Agents↗
PubMed1998

Decreased susceptibilities to teicoplanin and vancomycin among coagulase-negative methicillin-resistant clinical isolates of staphylococci.

Of 41 methicillin-resistant coagulase-negative staphylococcal clinical isolates collected during a 5-month period between late 1995 and early 1996, 28 showed tube dilution teicoplanin MICs of 4 to 8 microg/ml which increased to 16 to 32 microg/ml upon prolonged incubation. Cultures of such bacteria were heterogeneous; they contained subpopulations with frequencies of 10(-5) to 10(-4) that could grow on up to 50 microg of teicoplanin per ml. The same cultures were also heterogeneous with respect to susceptibility to vancomycin; while the MICs for the majority of cells were 2 to 4 microg/ml, subpopulations that could grow on 6 to 12 microg of vancomycin per ml were also present at frequencies of 10(-5) to 10(-7). Selective enrichment of such cultures for the resistant subpopulation occurred with relative ease under laboratory conditions. Heterogeneous phenotypes for teicoplanin (but not for vancomycin) susceptibility were also identified in several Staphylococcus epidermidis isolates collected during the preantibiotic era. The addition of half the MIC of teicoplanin inhibited autolysis and caused formation of cellular aggregates which disintegrated to individual bacteria in the stationary phase when the titer of teicoplanin in the medium fell to undetectable levels, indicating removal of the antibiotic from the culture medium by the bacteria.

Anti-Bacterial Agents↗
PubMed1998

Genetic organization of the mecA region in methicillin-susceptible and methicillin-resistant strains of Staphylococcus sciuri.

A homolog of the Staphylococcus aureus methicillin resistance gene mecA was recently shown to be ubiquitous in independent isolates of the animal species Staphylococcus sciuri. The mecA gene homolog and regions flanking it were cloned and sequenced from four strains of S. sciuri: strain K1 (ATCC 29062), a representative of S. sciuri subsp. sciuri; two strains (K3 and K8) representing S. sciuri subsp. rodentius; and strain K11, a representative of S. sciuri subsp. carnaticum. Strains K1 and K11 were susceptible to methicillin, while strains K3 and K8 showed heterogeneous resistance. The mecA genes of strains K1 and K11 and one of the two copies of mecA (mecA1) present in strain K3 had virtually identical DNA sequences in the mecA gene and were similar in genetic organization in the flanking regions. In contrast, the single copy of mecA in strain K8 and the second copy of mecA (mecA2) in strain K3 had mecA DNA sequences identical to that of S. aureus mecA, and the mecA region in these two strains was also similar to that of the same region in the S. aureus strain used for comparison. Interestingly, an open reading frame defining an N-terminal truncated polypeptide, NTORF101, with a high degree of homology to a DNA segment in the hypervariable region of methicillin-resistant S. aureus (and also similar to the Escherichia coli gene ugpQ) was also identified downstream of the mecA homolog of strain K11, representing S. sciuri subsp. carnaticum. The ugpQ-like gene is not present in methicillin-susceptible strains of S. aureus. The presence of such a ugpQ-like gene together with the homolog of mecA in strain K11 supports the speculation that these genetic elements may be evolutionary relatives and/or precursors of the genetic determinant of methicillin resistance in S. aureus.

Amino Acid Sequence↗
PubMed1998

Transcriptional analysis of the Staphylococcus aureus penicillin binding protein 2 gene.

Sequencing of the vicinity of the staphylococcal pbp2 gene and transcriptional analysis by primer extension and promoter fusions were used to show that pbp2 is part of an operon that also includes a gene with high homology to prfA of Bacillus subtilis. Two distinct promoters were identified directing transcription of pbp2 either alone or together with prfA. It was recently reported that transposon inactivation of pbp2 causes a reduction in methicillin resistance, but complementation experiments were not fully successful. We now show that introduction of the intact pbp2 gene with its two newly identified promoters into the chromosome of the transposon mutant resulted in the full recovery of high-level methicillin resistance.

Amino Acid Sequence↗
PubMed1998

Molecular epidemiology of penicillin-resistant Streptococcus pneumoniae isolates recovered in Italy from 1993 to 1996.

Thirty-nine penicillin-resistant Streptococcus pneumoniae isolates recovered among the approximately 700 pneumococcal strains collected from 1993 to 1996 in central and northern Italy were analyzed for several characteristics, including serotype, antibiotic susceptibility profile, chromosomal relatedness (by using pulsed-field gel electrophoresis [PFGE]), restriction fragment length polymorphism (RFLP) of the penicillin-binding protein (PBP) genes 1A, 2X, and 2B, and the presence of a variety of antibiotic resistance genes (determined by hybridization with appropriate DNA probes). The MICs of penicillin for most of the isolates (30 of 39) were high, in the range of 1 microgram/ml or higher, and these 30 isolates carried additional resistance traits to two or more drugs (erythromycin, chloramphenicol, co-trimoxazole, and tetracycline) and expressed serotypes 9, 19, and 23 and three distinct PFGE patterns. More than half (22 of 30) of the isolates for which MICs were high were identified as representatives of two widespread international epidemic clones of S. pneumoniae. The first one of these clones (seven isolates) expressed serotype 23F and possessed all properties characteristic of the widespread Spanish/USA international clone. Seven additional strains with serotype 19 also had the same PFGE pattern, PBP gene, and RFLP polymorphisms, and other properties typical of the serotype 23 Spanish/USA clone, suggesting that these strains were the products of a capsular transformation event (from serotype 23F to serotype 19) in which the Spanish/USA clone was the recipient. The second international clone was represented by eight serotype 9 isolates which were resistant to penicillin and trimethoprim-sulfamethoxazole and had the molecular properties of the French/Spanish epidemic clone. The remaining eight isolates for which penicillin MICs were high appeared to represent a hitherto-undescribed "Italian" clone; they had a novel PFGE type, unique RFLPs for the PBP genes, and resistance to tetracycline, trimethoprim-sulfamethoxazole, and erythromycin, and the penicillin MICs for these isolates were 2 to 4 microgram/ml.

Chromosome Mapping↗
PubMed1997

Structural characterization of an abnormally cross-linked muropeptide dimer that is accumulated in the peptidoglycan of methicillin- and cefotaxime-resistant mutants of Staphylococcus aureus.

Laboratory mutants of Staphylococcus aureus strain ATCC 8325 (27S) selected for increased minimal inhibitory concentration (MIC) values to methicillin and cefotaxime showed increased rates of cell wall turnover and detergent-induced autolysis in virtual parallel with the increasing MIC for the antibiotic. Also in parallel with the increasing MICs for the particular antibiotic used in the selection was the gradual accumulation of an unusual muropeptide in the peptidoglycan of the mutants, muropeptide 12, which is a minor component of the cell wall of the parental strain. Analysis of muropeptide 12, its peptide derivative, and its lysostaphin degradation products by high pressure liquid chromatography, Edman degradation, and mass spectrometry suggests that muropeptide 12 is a dimer in which the two monomeric components are interlinked by two pentaglycyl cross-bridges, thus generating a 14-member macrocyclic ring structure. This unusual cross-linked structure may be the product of the abnormal activity of penicillin-binding protein 2 which has grossly reduced antibiotic binding capacity in the mutant staphylococci.

Amino Acid Sequence↗
PubMed1997

Spread of the serotype 23F multidrug-resistant Streptococcus pneumoniae clone to South Korea.

Thirty-eight antibiotic-resistant isolates of Streptococcus pneumoniae recovered in a hospital in Seoul, Korea, between February 1990 and March 1992 were analyzed for serotype, antibiotic susceptibility patterns, and chromosomal relatedness using pulsed-field gel electrophoretic (PFGE) analysis of SmaI chromosomal digests. Most of the isolates were from sputum samples, and a few strains were from otitis media and meningitis. The great majority of isolates (34 of 38, or 89%) were multiresistant, sharing virtually identical, elevated minimal inhibitory concentration (MIC) values (microgram/ml) for penicillin (1-2), chloramphenicol (12-25), tetracycline (25-30), and sulfamethoxazole/trimethoprim (> 100). Twenty of the isolates were also resistant to erythromycin, and all isolates were also considered to be resistant to ceftriaxone and cefotaxime (1-2) according to the new breakpoint definitions. The most frequent serotypes were 23F (17 of 38) and 19F(14 of 38); 2 belonged to serotype 15B and 1 of 24F. Of the remaining 4 isolates (2 serotype 3, 1 type 6B, and 1 type 9V) all were resistant to tetracycline and sulfamethoxazole/trimethoprim and with the exception of 1 of the type 3 strains, were also resistant to chloramphenicol. Eleven of the 14 serotype 19F isolates shared a relatively homogeneous PFGE pattern, which was indistinguishable from the PFGE pattern shown by most (12 of 17) of the 23F isolates. The PFGE pattern of these 19F and 23F isolates was also indistinguishable from the PFGE pattern shown by representative multiresistant capsular type 23F isolates from Croatia, Portugal, and New York City and the findings document the extensive geographic spread of this multidrug-resistant S. pneumoniae clone. The data also suggests in vivo capsular transformation of the multiresistant clone from serotype 23F to serotype 19F.

DNA Fingerprinting↗
PubMed1997

Ubiquitous distribution of the competence related genes comA and comC among isolates of Streptococcus pneumoniae.

DNA probes specific for the pneumococcal competence regulatory genes comA and comC were used to test the presence of these genes in 214 pneumococcal isolates selected to represent a wide variety of clonal types and genetic backgrounds (as defined by chromosomal macrorestriction patterns and multilocus enzyme analysis), a variety of serotypes, isolation dates (between 1916 and 1996), geographic origins (on four continents), as well as different clinical origins (including both infection sites and colonization sites). Each isolate gave positive signal with both DNA probes. The comA and comC genes were never on the same SmaI restriction fragment and the comA gene showed a considerable degree of polymorphism from one strain to another. While DNA sequencing of 50 of the isolates have identified three distinct alleles of the comC gene, the number of mutations within the leader peptide were minimal. The results suggest that the mechanism to undergo genetic transformation is widespread within the species of Streptococcus pneumoniae.

Alleles↗
PubMed1997

Autolysis and cell wall degradation in a choline-independent strain of Streptococcus pneumoniae.

Streptococcus pneumoniae has an auxotrophic requirement for choline, and choline residues that incorporate into the wall and membrane teichoic acids are intimately involved with the control of autolytic phenomena of this bacterium. We report here the re-examination of the role of choline in autolytic cell wall degradation using the choline-independent S. pneumoniae strain R6Cho- recovered from a heterologous cross with DNA from Streptococcus oralis. S pneumoniae Cho- cultured in choline-free medium grew with normal generation time but formed long chains, failed to undergo stationary-phase autolysis, and was also resistant to lysis induced by deoxycholate or penicillin. Cell walls produced under these conditions had reduced phosphorus content, contained no choline residues detectable by nuclear magnetic resonance, and had reduced binding capacity for the pneumococcal autolytic amidase, and complete hydrolysis of such walls by the amidase required prolonged incubation with high concentrations of the enzyme. Addition of choline to the growth medium reversed at these phenomena. High-performance liquid chromatography analysis of amidase digests of cell walls prepared from strain R6Cho- grown with or without choline produced identical stem peptide profiles, which were also similar to that of the parental S. pneumoniae strain R6. Peptidoglycans prepared by hydrofluoric extraction of cell walls from Cho- growth with or without choline or from the parental strain R6 were uniformly susceptible to the autolytic amidase and were fully degraded to the normal family of stem peptides, indicating that, in sharp contrast to the case of cell walls, the amidase degradation of teichoic acid-free peptidoglycan did not require the presence of choline residues in the substrate.

Amidohydrolases↗