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PubMed1998

Combined inhibitory effect of bovine immune whey and peroxidase-generated hypothiocyanite against glucose uptake by Streptococcus mutans.

Immune whey product was obtained from Streptococcus mutans- and Streptococcus sobrinus-immunized cows. Hypothiocyanite (HOSCN/OSCN-) was generated in VMG-buffer, pH 5.5 or 6.5, by bovine milk lactoperoxidase, KSCN and hydrogen peroxide. The glucose incorporation by late-log cells of S. mutans 10449, serotype c, was followed by measuring the uptake of 14C-glucose at 37 degrees C. At pH 5.5 and 6.5 both immune whey product and HOSCN/OSCN- dose-dependently inhibited glucose uptake. The inhibition by their combination was additive if bacterial cells were treated with HOSCN/OSCN- before exposed to immune whey product. In contrast to immune whey product, the control product from sham-immunized cows increased the glucose uptake even when added simultaneously with HOSCN/OSCN-. However, when bacterial cells were pretreated with HOSCN/OSCN- an enhanced inhibitory effect was observed also with control product. The results indicate that colostral proteins from S. mutants- or S. sobrinus-immunized cows inhibit glucose uptake and that the effect is enhanced by pretreatment with lactoperoxidase-generated HOSCN/OSCN-.

Adjuvants, Immunologic↗
PubMed2005

Association of a novel high molecular weight, serine-rich protein (SrpA) with fibril-mediated adhesion of the oral biofilm bacterium Streptococcus cristatus.

The surface of the oral plaque bacterium Streptococcus cristatus is decorated with a lateral tuft of fibrils. The fibrillar tuft functions in the adhesion of S. cristatus to heterologous bacterial species in the plaque biofilm. The tuft typically consists of a densely packed fringe of shorter fibrils 238 +/- 19 nm long with longer, less abundant fibrils 403 +/- 66 nm long projecting through the fringe of short fibrils. The two types of fibrils in the tufts of S. cristatus have been refractory to biochemical separation, complicating their characterization. A hexadecane partition assay was used to enrich for subpopulations of S. cristatus CR311 (type strain NCTC 12479) having distinct fibrillar morphotypes. Negative staining in the TEM revealed that cells of a hydrophobic subpopulation of S. cristatus (CR311var1) carried only the long fibrils (395 +/- 32 nm). A hydrophilic subpopulation of S. cristatus (CR311var3) consisted of mixed morphotypes having no fibrils or remnant short fibrils (223 +/- 49 nm). No long fibrils were observed on any cells in the CR311var3 subpopulation. The CR311var3 morphotype, unlike the wild-type strain and CR311var1, was not able to form corncobs with either Corynebacterium matruchotii or Fusobacterium nucleatum. Variant CR311var3 did not express the novel gene srpA, which encodes a high molecular weight (321,882 Da) serine-rich protein, SrpA. The SrpA protein contains two extensive repeat motifs of 17 and 71 amino acids and a gram-positive cell wall anchor consensus sequence (LPNTG). The unusual properties of SrpA most closely resemble those of Fap1, the fimbrial-associated adhesin protein of Streptococcus parasanguis. The association of long fibrils, high surface hydrophobicity, ability to form corncob formations, and expression of the srpA gene suggest that SrpA is a long fibril protein in S. cristatus.

Amino Acid Sequence↗
PubMed1996

Cloning, sequencing and functional overexpression of the Streptococcus equisimilis H46A gapC gene encoding a glyceraldehyde-3-phosphate dehydrogenase that also functions as a plasmin(ogen)-binding protein. Purification and biochemical characterization of the protein.

We previously identified DNA sequences involved in the function of the complex promoter of the streptokinase gene from Streptococcus equisimilis H46A, a human serogroup C strain known to express this gene at a high level. As a prerequisite to understanding possible mechanisms that control the balance between the plasminogen activating and plasmin(ogen) binding capacities of H46A, we describe here its gapC gene encoding glyceraldehyde-3-phosphate dehydrogenase (GraP-DH, EC 1.2.1.12), a glycolytic enzyme apparently transported to the cell surface where it functions as a plasmin(ogen).binding protein. The gapC gene was cloned and sequenced and found to code for a 336-amino-acid polypeptide (approximately 35.9 kDa) exhibiting 94.9% sequence identity to the Plr protein from Streptococcus pyogenes shown by others to be capable of plasmin binding [Lottenberg, R., Broder, C. C., Boyle, M. D., Kain, S. J., Schroeder, B. L. & Curtiss, R. III (1992) J. Bacteriol. 174, 5204-5210]. To study the properties of the GapC protein, its gene was inducibly overexpressed in Escherichia coli from QIAexpress expression plasmids to yield the authentic GapC or (His)6GapC carrying a hexahistidyl N-terminus to permit affinity purification. Both proteins were functionally active, exhibiting specific GraP-DH activities of about 80 kat/mol (approximately 130 U/mg) after purification. Their binding parameters [association (ka) and dissociation (kd) rate constants, and equlibrium dissociation constants (Kd = kd/ka)] for the interaction with human Gluplasminogen and plasmin were determined by real-time biospecific interaction analysis using the Pharmacia BIAcore instrument. For comparative purposes, the commercial GraP-DH from Bacillus stearothermophilus (BstGraP-DH), a nonpathogenic organism, was included in these experiments. The Kd values for binding of plasminogen to GapC, (His)6GapC and BstGraP-DH were 220 nM, 260 nM and 520 nM, respectively, as compared to 25 nM, 17 nM and 98 nM, respectively, for the binding to plasmin. These data show that both the zymogen and active enzyme possess low-affinity binding sites for the gapC gene product and that the hexahistidyl terminus does not affect its function. Prior limited treatment with plasmin enhanced the subsequent plasminogen binding capacity of all three GraP-DHs, presumably by the exposure of new C-terminal lysine residues for binding to the zymogen.

Amino Acid Sequence↗
PubMed1991

Growth of Staphylococcus aureus and Streptococcus species in bovine mammary secretions during the nonlactating and peripartum periods following intramammary infusion of lipopolysaccharide at cessation of milking.

The objective was to determine if induced mammary inflammation at cessation of milking influenced growth of gram-positive mastitis pathogens in mammary secretions, particularly during early involution. Growth of all mastitis pathogens evaluated was similar in cell-free fat-free mammary secretions from LPS-infused and control glands. These data indicate that intramammary infusion of LPS at cessation of milking did not alter growth of gram-positive mastitis pathogens in mammary secretion during the nonlactating period. Stage of lactation and the nonlactating period influenced bacterial growth and marked differences between bacteria and among strains of a bacterial species were observed. Staphylococcus aureus grew well in secretions collected during late lactation, but growth decreased during early- and mid-involution and increased again in secretions obtained near parturition. Streptococcus agalactiae and Strep. uberis grew better in mammary secretion obtained during involution than in secretions collected during late or early lactation. Streptococcus dysgalactiae grew well in mammary secretions at all time periods. These data demonstrate the variability of mastitis pathogen growth during physiologic transitions of the bovine udder.

Animals↗
PubMed1998

Virulence factors of Streptococcus uberis isolated from cows with mastitis.

For intramammary infections (IMI) to occur, mechanisms associated with avoidance of phagocytic defenses, rapid growth of bacteria, adherence of bacteria to epithelial cells, and/or bacterial colonization of mammary tissue are probably present. During the last decade, several potential virulence factors of Streptococcus uberis have been identified. Some of these factors are cell-associated while other factors are extracellular. Proposed antiphagocytic factors of S. uberis include capsule, neutrophil toxin, M-like protein and R-like protein. Activation of plasminogen by S. uberis has been proposed as an important mechanism for this organism to obtain nutrients for optimal bacterial growth. Potential virulence factors produced by S. uberis and released extracellularly include hyaluronic acid capsule, hyaluronidase and uberis factor. Streptococcus uberis isolated from bovine IMI adhere to and invade mammary epithelial cells. Involvement of intact microfilaments and de novo eukaryotic protein synthesis are required for bacterial invasion of mammary epithelial cells; a process that appeared to occur by a receptor-mediated endocytosis mechanism. De novo bacterial protein synthesis was also required for invasion of S. uberis into mammary epithelial cells. Furthermore, S. uberis survived within mammary epithelial cells for extended periods of time without losing viability or damaging the eukaryotic cell. Further research directed towards characterization of host-pathogen interactions that take place during the early stages of S. uberis intramammary infection are needed to enhance our understanding of pathogenesis and thus contribute to development of methods to minimize production losses associated with S. uberis mastitis in dairy cows.

Animals↗
PubMed2006

Adherence and internalization of Streptococcus gordonii by epithelial cells involves beta1 integrin recognition by SspA and SspB (antigen I/II family) polypeptides.

Streptococcus gordonii is a commensal bacterium that colonizes the hard and soft tissues present in the human mouth and nasopharynx. The cell wall-anchored polypeptides SspA and SspB expressed by S. gordonii mediate a wide range of interactions with host proteins and other bacteria. In this article we have determined the role of SspA and SspB proteins, which are members of the streptococcal antigen I/II (AgI/II) adhesin family, in S. gordonii adherence and internalization by epithelial cells. Wild-type S. gordonii DL1 expressing AgI/II polypeptides attached to and was internalized by HEp-2 cells, whereas an isogenic AgI/II- mutant was reduced in adherence and was not internalized. Association of S. gordonii DL1 with HEp-2 cells triggered protein tyrosine phosphorylation but no significant actin rearrangement. By contrast, Streptococcus pyogenes A40 showed 50-fold higher levels of internalization and this was associated with actin polymerization and interleukin-8 upregulation. Adherence and internalization of S. gordonii by HEp-2 cells involved beta1 integrin recognition but was not fibronectin-dependent. Recombinant SspA and SspB polypeptides bound to purified human alpha5beta1 integrin through sequences present within the NAV (N-terminal) region of AgI/II polypeptide. AgI/II polypeptides blocked interactions of S. gordonii and S. pyogenes with HEp-2 cells, and S. gordonii DL1 cells expressing AgI/II proteins inhibited adherence and internalization of S. pyogenes by HEp-2 cells. Conversely, S. gordonii AgI/II- mutant cells did not inhibit internalization of S. pyogenes. The results suggest that AgI/II proteins not only promote integrin-mediated internalization of oral commensal streptococci by host cells, but also potentially influence susceptibility of host tissues to more pathogenic bacteria.

Adhesins, Bacterial↗
PubMed2006

Clinical implications and treatment of multiresistant Streptococcus pneumoniae pneumonia.

Streptococcus pneumoniae is the leading bacterial cause of community-acquired respiratory tract infections. Prior to the 1970s this pathogen was uniformly susceptible to penicillin and most other antimicrobials. However, since the 1990s there has been a significant increase in drug-resistant Streptococcus pneumoniae (DRSP) due, in large part, to increased use of antimicrobials. The clinical significance of this resistance is not definitely established, but appears to be most relevant to specific MICs for specific antimicrobials. Certain beta-lactams (amoxicillin, cefotaxime, ceftriaxone), the respiratory fluoroquinolones, and telithromycin are among several agents that remain effective against DRSP. Continued surveillance studies, appropriate antimicrobial usage campaigns, stratification of patients based on known risk factors for resistance, and vaccination programmes are needed to appropriately manage DRSP and limit its spread.

Anti-Infective Agents↗
PubMed2006

Antibiotic consumption and generation of resistance in Streptococcus pneumoniae: the paradoxical impact of quinolones in a complex selective landscape.

The development of resistance to the different antibiotics by the majority of bacterial species of clinical importance seems unavoidable. However, not all drugs have the same efficiency to select for resistance. Large differences in the qualitative and quantitative consumption of antibiotics among countries are known to exist and several authors have consistently reported the direct relationship between consumption and selection of resistance for Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and Escherichia coli and beta-lactams and macrolides use. In Spain, extensive surveillance started in 1996, and the Willow (SAUCE) Project, to monitor and update resistance in respiratory pathogens and to couple those data with data concerning national antibiotic consumption (IMS) from both a temporal and geographical approach. Temporally, despite a continuous increase of 16% in quinolone consumption from 1997 to 2001, basically due to the arrival of respiratory quinolones, levofloxacin and moxifloxacin, a continuous linear increase in the resistance rates to ciprofloxacin in S. pneumoniae was not observed. There also was an inverse correlation between provincial consumption of quinolones and resistance to ciprofloxacin. Several hypotheses are proposed and discussed to explain these apparent paradoxical observations, such as the replacement of ciprofloxacin by more potent antipneumococcal quinolones, the possibility of an antibiotic pressure threshold, the influence of other nonquinolone drugs on the expression of ciprofloxacin-resistance biological costs, and the influence of changes in temporal or spatial prevalence of particular clones.

Anti-Bacterial Agents↗
PubMed1994

Antepartum screening and non-selective intrapartum chemoprophylaxis for group B streptococcus.

The efficacy of antibiotic chemoprophylaxis in labour for all maternal carriers of Group B streptococcus with respect to the neonatal mortality and morbidity was analyzed. The intervention consisted of a policy of maternal screening for Group B streptococcus at 28 weeks' gestation by a low vaginal swab, cultured on to nonselective media. If positive, all carriers were treated with intrapartum ampicillin regardless of other obstetric risk factors. The main outcome measures were studied prospectively for 16 months before and for 32 months after instituting the intervention. These included the incidence of early-onset Group B streptococcal infection as defined by either a blood culture or a symptomatic neonate with a positive urinary streptococcal antigen test, the mortality and the morbidity. Intrapartum ampicillin significantly reduced the incidence of early-onset Group B streptococcal disease, from 2.0 to 0.37 per 1,000 livebirths for neonates with a positive blood culture (p < 0.03), and from 4.9 to 1.5 per 1,000 livebirths for symptomatic infants with a positive urinary antigen test (p < 0.004) and the attributable morbidity, namely, neonates who were: admitted to a level 2 or 3 unit (p < 0.004); treated with antibiotics (p < 0.004); preterm (p < 0.04); ventilated (p < 0.01). Seventy five per cent of the maternal population was overall actually screened and the carriage rate was 12%. No serious adverse reactions to ampicillin were observed. Significant reduction in early-onset neonatal Group B streptococcal disease has occurred concurrent with antepartum screening and nonselective intrapartum chemoprophylaxis.

Ampicillin↗
PubMed1998

Parenteral vs oral antibiotics in the prevention of serious bacterial infections in children with Streptococcus pneumoniae occult bacteremia: a meta-analysis.

OBJECTIVE: To determine whether parenteral antibiotics are superior to oral antibiotics in preventing serious bacterial infections in children with Streptococcus pneumoniae occult bacteremia. METHODS: Using the MEDLINE database, the English language literature was searched for all publications concerning bacteremia, fever, or Streptococcus pneumoniae from 1966 to January 1, 1997. All nonduplicative studies with a series of children with S. pneumoniae occult bacteremia having both orally treated and parenterally treated groups were reviewed. Children were excluded from individual studies if at the time of their initial evaluation they were immunocompromised, had a serious bacterial infection, underwent a lumbar puncture, or did not receive antibiotics. RESULTS: Only 4 studies met study criteria. From these studies, 511 total cases of S. pneumoniae occult bacteremia were identified. Ten of 290 (3.4%) in the oral group and 5 of 221 (2.3%) in the parenteral antibiotic group developed serious bacterial infections (pooled p-value = 0.467, pooled OR = 1.48; 95% CI, 0.5-4.3). Two patients in the oral group (0.7%) and 2 patients in the parenteral group (0.9%) developed meningitis (pooled p-value = 0.699, pooled OR = 0.67; 95% CI, 0.1-5.1). CONCLUSION: The rates of serious bacterial infections and meningitis did not differ between children who were treated with oral and parenteral antibiotics. The extremely low rate of complications observed in both groups suggests no clinically significant difference between therapies. A study with >7,500 bacteremic children (or >300,000 febrile children) would be needed to have 80% power to prove parenteral antibiotics are superior to oral antibiotics in preventing serious bacterial infections.

Administration, Oral↗
PubMed2006

The effect of ingestion of milk supplemented with salivaricin A-producing Streptococcus salivarius on the bacteriocin-like inhibitory activity of streptococcal populations on the tongue.

The colonization efficacies of salivaricin A (SalA)-producing Streptococcus salivarius strains 20P3 and 5 were compared when given in milk to 219 children, using either 2-day or 9-day dosing regimens. Colonization levels overall were superior for strain 5, and the 9-day dosing schedule resulted in higher levels of both initial colonization and strain persistence. The indigenous streptococcal tongue populations of 20 (10.9%) of the 189 children in the 2-day trial showed markedly increased SalA-like inhibitory activity following use of the S. salivarius-supplemented milk. All 20 of these children were found to have had relatively small (<5% of total S. salivarius) indigenous tongue populations of SalA-producing S. salivarius, and the relative proportions and/or inhibitory activity of these SalA producers on the childrens' tongues increased following ingestion of the S. salivarius-supplemented milk. Because SalA is known to be strongly inhibitory to Streptococcus pyogenes, an important implication of this study is that the consumption of SalA-producing probiotic S. salivarius could potentially help to effect a sustained increase in SalA-mediated protection against S. pyogenes infection.

Animals↗
PubMed1995

Partial characterization of the cohemolytic factor produced by Streptococcus uberis and comparison with the CAMP-factor.

Exosubstances (cohemolysins) produced by Streptococcus agalactiae (CAMP-factor) and Streptococcus uberis (Uberis-factor) showing hemolytic synergism with beta-lysin produced by Staphylococcus aureus were compared. Cohemolytic activity was evaluated in the supernatants of bacterial cultures, before and after ammonium sulfate precipitation. Sheep erythrocytes sensitized with beta-lysin were used as substrate. The assays were performed in microtiter plates and results were expressed as cohemolytic units/ml. Maximum cohemolytic activity was detected, respectively, after 8 h and 14 h of growth in Columbia broth in S. uberis and S. agalactiae cultures. Cohemolytic activities of both microorganisms showed similarities when submitted to various physical and chemical treatments. They were significantly decreased by heating at 60 degrees C and 100 degrees C, or in presence of trypsin, and were abolished in the presence of Tween 20. Activities were found to be stable in crude supernatants and concentrated preparations maintained at -20 degrees C for 3 months. Differences were related to levels of activity and kinetics of detection during the growth cycle. The results indicate the proteic nature, at least in part, of the Uberis factor. Analysis by PAGE in the presence or absence of SDS allowed us to correlate Uberis activity with a protein band with apparent molecular mass of 42 kDa, while CAMP activity was associated with a protein band of 27 kDa.

Bacterial Proteins↗
PubMed1999

Immunological cross-reactivity between the vaccine and other isolates of Streptococcus bovis and Lactobacillus.

Recent studies have showed that immunisation with Streptococcus bovis (Sb-5) and Lactobacillus (LB-27) may confer protection against lactic acidosis in sheep and cattle. The present study was designed to determine the degree of immunological cross-reactivity between Sb-5 and eight other strains of Streptococcus bovis; and between LB-27 and four other isolates of Lactobacillus. The cross-reactivity index (CRI, a low CRI indicates a high degree of immunological cross-reactivity) ranged from 7.3 to 56.1% between the strains of S. bovis (the encapsulated strains with CRIs ranging from 7.3 to 12.4%). For isolates of Lactobacillus the CRIs ranged from 11.5 to 72.2%. The results indicate that all the isolates tested have a certain degree of immunological homology with Sb-5 and LB-27, and suggest that the vaccine may cross-react with a large number of strains of S. bovis and Lactobacillus which may cause lactic acidosis. As most of the S. bovis strains in the rumen are encapsulated, the high degree of homology between Sb-5 and encapsulated S. bovis strains further suggests that the vaccine containing Sb-5 may be effective against a wide range of strains of S. bovis in sheep and cattle.

Animals↗
PubMed1993

Activation of bovine plasminogen by Streptococcus uberis.

Culture filtrate from Streptococcus uberis was found to activate bovine and equine plasminogen but not that from rabbit, human or porcine plasma. In contrast, streptokinase from a Lancefield group C Streptococcus activated human plasminogen but not that from bovine, porcine and rabbit plasma. Very slight activity was observed against equine plasminogen. Plasmin was detected by hydrolysis of skimmed milk protein in agarose. The activation of bovine plasminogen by S. uberis culture filtrate resulted in the formation of three polypeptides with molecular masses of 56, 26 and 21 kDa. This is the first report of a streptokinase activity from this species.

Animals↗
PubMed1994

Fluoride inhibits the glucan-binding lectin of Streptococcus sobrinus.

The glucan-binding lectins of Streptococcus cricetus AHT and Streptococcus sobrinus 6715 were reversibly inhibited by sodium fluoride. Fluoride was superior to chloride, bromide, iodide and thiocyanate in preventing glucan-mediated aggregation of the bacteria. Fluoride was also an effective inhibitor of the sucrose-dependent adhesion of S. sobrinus to glass surfaces. The inhibition of glucan-binding lectin activities may be one of the mechanisms of action of fluoride in preventing dental disease.

Bacterial Adhesion↗
PubMed1997

Nucleotide sequence and molecular characterization of a gene encoding GTP-binding protein from Streptococcus gordonii.

A 1286-bp fragment of chromosomal DNA from Streptococcus gordonii strain Challis was cloned and sequenced. The gene sgg consisted of 897-bp nucleotides encoding a 299-amino acid polypeptide (33,200 Da). The deduced amino acid sequence exhibited significant similarity to Era, G protein of Escherichia coli. The nucleotide binding assay demonstrated that recombinant Sgg bound [32P]GTP but not [32P]ATP, [32P]CTP, or [32P]UTP. These findings indicate that Sgg is a member of the G protein superfamily in the genus Streptococcus.

Amino Acid Sequence↗
PubMed1998

Zoocin A immunity factor: a femA-like gene found in a group C streptococcus.

A 6.8-kb fragment of Streptococcus equi subsp. zooepidemicus 4881 DNA containing the zoocin A gene (zooA) was cloned in Escherichia coli and sequenced. We have identified a gene we call zoocin A immunity factor (zif), which protects the producer cell from the otherwise lethal action of its own product. Transformation of Streptococcus gordonii DL1 with zooA and zif changed its phenotypic character from a non-zoocin A producing-zoocin A sensitive cell to a zoocin A producing-zoocin A resistant cell. zif has sequence homology to femA (factor essential for methicillin resistance) and lif (lysostaphin immunity factor). No differences were observed in amino acid or amino sugar compositions of peptidoglycan purified from zoocin A sensitive vs. zoocin A immune cells.

Amino Acids↗
PubMed1998

Physical and genetic chromosomal maps of Streptococcus agalactiae, serotypes II and III; rRNA operon organization.

A detailed analysis of two Streptococcus agalactiae (group B streptococcus, GBS) strains was performed by pulsed field gel electrophoresis (PFGE). Digestion of the chromosomal DNA with SmaI and SgrAI endonucleases, followed by separation and analysis of fragments by PFGE was carried out. Physical chromosomal maps of serotype II/(alpha + beta) and III/alpha strains of S. agalactiae were constructed. The GBS genome size was estimated to be 2200 kb. Sixteen GBS genes were used as probes and were located on the restriction maps of both strains by DNA-DNA hybridization. Six copies of ribosomal operons were found in the genome of the analyzed strains. Significant differences in the restriction patterns of chromosomal DNA and DNA-DNA hybridization between the two analyzed strains were detected so that DNA restriction patterns may be used to trace outbreaks of disease. The overall GBS chromosomal organization as determined is fairly conserved.

Base Sequence↗