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PubMed1998

Characterisation of the gene encoding suilysin from Streptococcus suis and expression in field strains.

The gene encoding suilysin was cloned from Streptococcus suis serotype 2 strain P1/7. Analysis of the nucleotide and translated amino acid sequence confirmed suilysin to be a member of the thiol activated cytolysin group (TACY). The pneumolysin from Streptococcus pneumoniae is the most closely related orthologous gene known. Suilysin was overexpressed in E. coli in an active haemolytic form. A strong correlation between the presence of the sly gene and haemolytic activity in the supernatant of S. suis field strains was found. Of 158 strains tested, 63% contained the gene. Within the (most prevalent) serotype 2, the sly gene was demonstrated in 95% of the strains isolated in Eurasia, but only in 7% of the strains from North America.

Animals↗
PubMed2000

Shifts in membrane fatty acid profiles associated with acid adaptation of Streptococcus mutans.

Cells of Streptococcus mutans UA159 physiologically adapted to acidification during growth at pH 5 in glucose-limited chemostat cultures were enriched in mono-unsaturated and longer chain fatty acids compared with unadapted cells grown under the same conditions but at pH 7. Ratios of unsaturated to saturated fatty acids in the cells were, respectively, 1.2 and 0.3. Cyclopropane fatty acids were not detected. Streptococcus sobrinus 6715, which is known to have minimal acid-adaptive capacity, showed only minimal change in membrane fatty acids.

Adaptation, Physiological↗
PubMed2002

Screening of large numbers of Streptococcus pneumoniae isolates for mutations associated with fluoroquinolone resistance using an oligonucleotide probe assay.

Mutations at a relatively small number of sites in parC, parE and gyrA account for most of the fluoroquinolone resistance in Streptococcus pneumoniae clinical isolates. A high throughput oligonucleotide probe assay was developed to screen for mutations in the quinolone-resistance determining region (QRDR) of parC (Ser79), gyrA (Ser81) and parE (Asp435) of Streptococcus pneumoniae. Eight oligonucleotide probes (17mers) were used in the presence of tetramethyl ammonium chloride so that the melting temperature was dependent on length and not on base composition. Using this assay it was possible to accurately detect QRDR mutations from several hundred S. pneumoniae clinical isolates that were grown on nylon membranes.

Anti-Infective Agents↗
PubMed2006

Presence of NAD+-specific glyceraldehyde-3-phosphate dehydrogenase and CcpA-dependent transcription of its gene in the ruminal bacterium Streptococcus bovis.

To know the role of NADP+-specific glyceraldehyde-3-phosphate dehydrogenase (GAPN) in Streptococcus bovis, the molecular properties and transcriptional control of the gene encoding GAPN (gapN) were examined. The GAPN in S. bovis was deduced to consist of 476 amino acids with a molecular mass of 51.1 kDa. The gapN gene was transcribed in a monocistronic fashion. GAPN synthesis appeared to be regulated at the transcriptional level in response to changes in growth conditions. In a mutant that lacks the ccpA gene encoding catabolite control protein A (CcpA), the gapN-mRNA level was lower than in the parent strain. A binding site of CcpA was found in the upper region of gapN. These results suggest that transcription of gapN is regulated through CcpA. Overexpression of GAPN in S. bovis did not affect the growth rate or formate-to-lactate ratio, suggesting that the flux in the glycolytic pathway is unlikely to be altered by GAPN activity. Streptococcus bovis GAPN was NADP+ dependent, but not phosphate dependent. In addition, S. bovis did not have other NADPH-producing systems such as the hexose monophosphate pathway and NADPH:NAD+ oxidoreductase. Therefore, GAPN may play an important role in NADPH production in S. bovis.

Animals↗
PubMed2006

Inhibitory effects of cranberry polyphenols on formation and acidogenicity of Streptococcus mutans biofilms.

Cranberry fruit is a rich source of polyphenols, and has shown biological activities against Streptococcus mutans. In the present study, we examined the influence of extracts of flavonols (FLAV), anthocyanins (A) and proanthocyanidins (PAC) from cranberry on virulence factors involved in Streptococcus mutans biofilm development and acidogenicity. PAC and FLAV, alone or in combination, inhibited the surface-adsorbed glucosyltransferases and F-ATPases activities, and the acid production by S. mutans cells. Furthermore, biofilm development and acidogenicity were significantly affected by topical applications of PAC and FLAV (P<0.05). Anthocyanins were devoid of any significant biological effects. The flavonols are comprised of mostly quercetin glycosides, and the PAC are largely A-type oligomers of epicatechin. Our data show that proanthocyanidins and flavonols are the active constituents of cranberry against S. mutans.

Anthocyanins↗
PubMed2006

Identification and characterization of the loci encoding the competence-associated alternative sigma factor of Streptococcus gordonii.

In naturally-competent streptococci such as Streptococcus pneumoniae, expression of the late competence operons is regulated by ComX (sigma(X)), the competence-specific alternative sigma factor. In this study, duplicate genes (comR1 and comR2) encoding the putative ComX homologue of the oral bacterium Streptococcus gordonii were identified. Like the identical twin comX loci of S. pneumoniae, both comR determinants are independently functional as well as responsive to the ComDE signal transduction system activated by competence-stimulating peptide. However, in contrast to the comX system, nucleotide sequence analyses in combination with in trans complementation studies with a comR null mutant demonstrate that the identical 83 bp tracts (Region I) located immediately upstream of the comR structural genes are insufficient to confer wild-type competence levels. Wild-type transformation levels required additional distal nonhomologous DNA segments (Region II). Our findings suggest that alternative regulatory elements, under overall control of the ComDE pathway, may influence expression of the comR loci.

Bacterial Proteins↗
PubMed2006

Occurrence of Streptococcus macedonicus in Italian cheeses.

A new approach for the detection and enumeration of Streptococcus macedonicus in cheese was developed. The method which is based on a first screening of cheeses by a PCR assay specific for S. macedonicus followed by plating positive samples on a differential medium (SM medium) was applied to 51 samples derived from PDO and traditional Italian cheeses. Streptococcus macedonicus was found in 16 of the 51 samples examined in the present work. With the exclusion of an Asiago cheese sample in which very high numbers of S. macedonicus (7.13 log CFU g(-1)) were found, the counts of S. macedonicus in SM medium ranged from 2.48 to 4.70 log CFU g(-1). In the same cheeses, total streptococci enumerated onto M17 agar were found at higher concentrations with values up to 7.88 log CFU g(-1). The system developed was particularly useful for the differential count of S. macedonicus in cheese and allowed to evaluate the occurrence of this species within the complex microbial lactic acid bacteria (LAB) population, which is typical of traditional cheeses. Results showed that in the examined cheeses S. macedonicus cannot be considered as a dominant LAB species.

Cheese↗
PubMed2006

IdeE, an IgG-endopeptidase of Streptococcus equi ssp. equi.

Streptococcus equi ssp. equi is the causative agent of strangles, a highly contagious and serious disease in the upper respiratory tract of horses. The present study describes the characterization of IdeE, a homolog of the secreted IgG-specific protease IdeS/Mac of Streptococcus pyogenes. The activity of IdeE is compared with the activity of IdeZ, the corresponding enzyme of the closely related S. equi ssp. zooepidemicus. A study of the proteolytic activity of recombinant IdeE and IdeZ on IgG from a selection of mammals shows that only antibodies containing the substrate site of IdeS/Mac are cleaved, indicating that the specificities of these enzymes are similar. Interestingly, IgG from horse is less effectively cleaved than IgG from e.g. dog or humans, as the dominating IgG isotype in horse sera (IgG4) lacks a distinct substrate site for IdeE/IdeZ. IgG-degradation is observed when S. equi ssp. equi is grown in the presence of horse serum, but not when grown with purified IgG. As the fraction of degraded IgG contains IgG4, the observed activity might be due to the expression of an unknown enzyme rather than IdeE. In a similar assay, no proteolysis of IgG was detected in the growth media of S. equi ssp. zooepidemicus.

Animals↗
PubMed2006

Molecular serotype identification of Streptococcus agalactiae of bovine origin by multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay.

We used a multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay and sequencing of a variable region of the cps cluster to identify serotypes of 140 Streptococcus agalactiae (group B Streptococcus; GBS) isolates from cattle. Only 71 (51%) isolates were typeable using antisera, but molecular serotypes (MS) were assigned to 133 (95%) and 139 (99%) isolates by partial cpsE-cpsF-cpsG sequencing and mPCR/RLB, respectively. Ninety-four isolates (67%) belonged to MS III and most belonged to a molecular serosubtype (msst) III-3, which is uncommon among GBS isolates from humans. Our results demonstrate that cps clusters of bovine GBS differ significantly from those of GBS isolates from humans.

Animals↗
PubMed2006

Establishing a genetic system for ecological studies of Streptococcus oligofermentans.

Streptococcus oligofermentans is a newly characterized species belonging to the mitis group of oral streptococci. So far no correlation has been demonstrated between S. oligofermentans and dental caries. Furthermore, a reverse correlation has been observed between the number of S. oligofermentans and the number of Streptococcus mutans, a major cariogenic pathogen, in the oral cavity. These properties suggest that S. oligofermentans may have a potential to be used as a 'probiotics' for caries prevention. In this study, we aim to establish a genetic system in S. oligofermentans to further study the biology of this new species. Using homologous regions of the comCDE locus in other streptococci, the comC gene was isolated and sequenced. A synthetic competence-stimulating peptide (CSP) was synthesized and shown to be able to effectively induce competence in S. oligofermentans. This CSP-induced transformation system in S. oligofermentans was used to construct green fluorescent protein (gfp) and luciferase (luc) reporter systems, both of which are driven by the lactate dehydrogenase (ldh) promoter. These reporter systems were further shown to be highly expressed in planktonic and biofilm cells, suggesting that these reporter systems can be used in future ecological studies of S. oligofermentans.

Bacterial Proteins↗
PubMed2007

Genetic variation in comC, the gene encoding competence-stimulating peptide (CSP) in Streptococcus mutans.

The genetic variability in comC, the gene encoding the quorum-sensing molecule, competence-stimulating peptide (CSP) in Streptococcus mutans is reported. Seven comC alleles encoding three distinct mature CSPs were identified among 36 geographically diverse strains, although, compared with Streptococcus pneumoniae, the amount of predicted amino acid sequence variation is low. In agreement with other studies, significant variation was found in the natural competence for DNA uptake in these strains. However, there was no correlation between the CSP genotype and the ability to transform these strains. Representative strains encoding each of the CSP variants became competent in response to synthetic CSPs of each type. Therefore, in contrast to S. pneumoniae, comC alleles in S. mutans are functionally equivalent and there is no evidence of pherotype specificity.

Alleles↗
PubMed2005

Design of a multiplex PCR for Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae and Chlamydophila pneumoniae to be used on sputum samples.

A multiplex PCR (mPCR) was developed for simultaneous detection of specific genes for Streptococcus pneumoniae (lytA), Mycoplasma pneumoniae (P1), Chlamydophila pneumoniae (ompA), and Haemophilus influenzae (16S rRNA, with verification PCR for P6). When the protocol was tested on 257 bacterial strains belonging to 37 different species, no false negatives and only one false positive were noted. One Streptococcus mitis out of thirty was positive for lytA. In a pilot application study of 81 sputum samples from different patients with suspected lower respiratory tract infection (LRTI), mPCR identified S. pneumoniae in 25 samples, H. influenzae in 29, M. pneumoniae in 3, and C. pneumoniae in 1. All samples culture positive for S. pneumoniae (n=15) and H. influenzae (n=15) were mPCR positive for the same bacteria. In a pilot control study with nasopharyngeal swabs and aspirates from 10 healthy adults, both culture and mPCR were negative. No PCR inhibition was found in any of the mPCR-negative sputum or nasopharyngeal samples. Whether all samples identified as positive by mPCR are truly positive in an aetiological perspective regarding LRTI remains to be evaluated in a well-defined patient material. In conclusion, the mPCR appears to be a promising tool in the aetiological diagnostics of LRTI.

Adolescent↗
PubMed1995

Failure of cefotaxime treatment in two children with meningitis caused by highly penicillin-resistant Streptococcus pneumoniae.

Two infants, aged 8.5 and 11 months, were admitted for meningitis caused by Streptococcus pneumoniae. Failure of cefotaxime led to the identification of highly penicillin-G-resistant strains. Minimum inhibitory concentrations (MICs) for penicillin were > 2 micrograms/ml, and cefotaxime MICs were 2 micrograms/ml. Both patients rapidly responded to a combination of i.v. imipenem and rifampicin. It is now mandatory to test in-vitro susceptibilities of Streptococcus pneumoniae to penicillin G and other beta-lactam agents when meningitis is diagnosed in infants.

Anti-Bacterial Agents↗
PubMed1990

Early response in neonatal septicemia. The effect of Escherichia coli, Streptococcus agalactiae and tumor necrosis factor on the generation of lactoferrin.

Using an in vitro model, we report the early effect of Escherichia coli (E. coli), Streptococcus agalactiea (group B streptococci, GBS) and recombinant tumor necrosis factor alpha (TNF) on the release of lactoferrin (LF) and the generation of interleukin-1 (IL-1) due to E. coli, using heparinized whole blood from healthy full-term newborns. We wanted to find a dose response relationship between lactoferrin generation on the one hand and the amount of E. coli, GBS and TNF on the other hand. In a final concentration of 10(7) per ml both bacteria increased the release of LF significantly. The response to E. coli was immediate and mg/l +/- 0.29 mg/l, E. coli 1.83 mg/l +/- 0.76 mg/l, p less than 0.001). GBS was a less potent stimulant than E. coli and the response was only apparent after 20 minutes (mean +/- S.D.: 1.06 mg/l +/- 0.49 mg/l, p less than 0.01). TNF in a concentration of 10000 pg/ml as well as 1000 pg/ml increased the release of LF significantly (after 20 minutes mean +/- S.D.: 1.09 mg/l +/- 0.42 mg/l and 0.97 mg/l +/- 0.36 mg/l, respectively), whereas a concentration of 100 pg/ml had no effect. Whole blood incubated with different preparations of LF for 20 minutes did not increase the generation of LF significantly. No significant changes in IL-1 levels were observed. Lactoferrin had bacteriostatic but no bactericidal effect on GBS and Streptococcus mutans.

Animals↗
PubMedDate not supplied

Caries experience, periodontal treatment needs, salivary pH, and Streptococcus mutans counts in a preadolescent Down syndrome population.

The prevalence of dental caries and periodontal treatment needs in an institutionalized population with Down syndrome was examined and the relationship between caries prevalence, salivary pH, and salivary levels of Streptococcus mutans was studied. Thirty-two children with Down syndrome, aged 8 to 13, were compared with two control groups of similar age ranges: healthy children and non-Down mentally retarded (MR) children living in the same institution as the Down syndrome population. The gender ratio mixtures in each group also matched the study sample. Caries experience as indicated by decayed, missing, and filled surfaces (DMF-S) showed significantly lower mean scores for the Down syndrome group compared with both control groups. A similar pattern was found when evaluating the mean numbers of decayed surfaces. Streptococcus mutans counts, expressed as number of colony-forming units on mitis salivarius agar plates among the Down syndrome group, were the lowest, although not statistically significant compared with the counts of the healthy children. Both groups had bacterial counts which were significantly lower than those of the MR group. Significant differences between the two institutionalized groups and the healthy group were recorded for the salivary pH levels. The periodontal treatment needs as evaluated by the Community Periodontal Index of Treatment Needs showed significant difference between the MR group versus both the healthy population and the Down syndrome group. On the whole, 84% of the Down syndrome children were cariesfree.

Adolescent↗
PubMed1989

[A case of infective endocarditis due to Streptococcus agalactiae].

A 60 year-old woman, who was pointed out as having hyperglycemia, was admitted to our hospital, with a two-week history of fever. Blood sugar on admission was 424 mg/dl and blood cultures yielded Streptococcus agalactiae. Systolic and diastolic heart murmurs were heard. Echocardiography showed aortic regurgitation and vegetation on the aortic valve. Penicillin-G (4 million units every 4 hr) iv and streptomycin (1 g every 12 hr) im were started. Chemotherapy was continued for 48 days. After chemotherapy, aortic valve replacement was done. We reported a very rare infective endocarditis case due to Streptococcus agalactiae.

Endocarditis, Bacterial↗
PubMed1989

[A study of the antigenicity of group A streptococcus strain "Matsuyama 2166" and its prevalence].

Antigen analysis of group A streptococcus strain, "Matsuyama 2166", which had been typed as T12-28 (by the T-agglutination method), MNT (nontypeable by the M-precipitation method) using conventional typing sera was examined by the precipitation and precipitation absorption technique. The prevalence of group A streptococci, typed under the name "Matsuyama 2166" was also investigated. The results were as follows: 1) The strain "Matsuyama 2166" is OF (Opacity Factor) (+) and its serological components consist of M "Matsuyama 2166" antigen, which is unique in our collection of M-types, and T28 antigen as a major antigen along with T12 as a minor. 2) Out of the group A streptococci typed as MNT, 96.6% of the streptococci were types as T12-28 and 96.5% of T28 by conventional typing sera were strains to be typed as M "Matsuyama 2166" using the newly prepared M "Matsuyama 2166" typing serum, suggesting the great advantage to M-typing rate of T28 strains. 3) Group A streptococcus M "Matsuyama 2166" was also found in isolates from Thailand or Malaysia. It is interesting that T-types found in those isolates seems to be a little different from the T-types isolated in Japan. These results showed that there was the difficult problem when we did the speculation of M-type from the result of T-type.

Antigens, Bacterial↗
PubMed1994

[Three interesting pediatric cases with penicillin-resistant Streptococcus pneumoniae infection].

Since penicillin-resistant Streptococcus pneumoniae first recognized in 1967, the rate of penicillin-resistant strains has been increasing worldwide. There have been up to 50% from pediatric specimens in Japan. We reported three pediatric cases with penicillin G resistant Streptococcus pneumoniae infection to show some important clue from these cases for clinical practice against resistant pneumococcal infection. The first case was a typical acute mastoiditis, although we have experienced only masked mastoiditis recently. The second case was meningitis with septicemia, which did not show any abnormality in the first obtained cerebrospinal fluid. The third case was recurrent bronchitis in a child with cerebral palsy. The minimum inhibition concentrations of these isolated strains were 0.25 microgram/ml in the second case an 2.0 microgram/ml in the first and third cases.

Adolescent↗