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Interaction between human polymorphonuclear leukocytes and Streptococcus milleri group bacteria.

Because Streptococcus milleri group (SMG) bacteria--Streptococcus constellatus, Streptococcus intermedius, and Streptococcus anginosus--exhibit a striking propensity to cause abscesses, the interaction of these organisms with human polymorphonuclear leukocytes (PMNL) was examined. After incubation in pooled normal human serum, SMG stimulated less chemotaxis than did Staphylococcus aureus, in contrast to viridans streptococci, which caused greater chemotaxis than did S. aureus. PMNL ingested greater numbers of SMG and viridans streptococci than S. aureus but killed these organisms more slowly and less completely. Relative resistance to killing by PMNL is expected in organisms that cause abscesses, and inhibition of chemotaxis may contribute to pathogenicity, because delayed arrival of PMNL gives a head start to proliferating bacteria. This study helps explain the capacity of SMG to cause abscesses. It is unclear, however, why viridans streptococci, bacteria that rarely produce abscesses, share some of these same properties.

Abscess↗

Group A Streptococcus septicemia and an infected, ruptured abdominal aortic aneurysm associated with pharyngitis.

A 65-year-old man had a 3-day history of sore throat, fever, rigors, back pain, abdominal discomfort, nausea, vomiting, and diarrhea. The patient's daughter had group A streptococcus pharyngitis. The patient was found to have a ruptured abdominal aortic aneurysm. He underwent resection of the aneurysm and right axillary femoro-femoral bypass graft. The patient died 40 hours after admission. Gram stain of the aneurysm showed numerous gram-positive cocci. Group A streptococcus grew from cultures of blood, throat, and aneurysm. The group A streptococcus was M type 3, T type 3 and produced streptococcal pyrogenic exotoxin A. This case is a very rare fatal complication of group A streptococcus pharyngitis.

Aged↗

Intrapartum transmission of group A streptococcus.

Intrapartum transmission of group A streptococcus has not been well documented. As the incidence of severe infections due to this organism has recently increased, it is important to assess if such transmission occurs. We observed two cases of severe neonatal infections due to group A streptococcus, one of which was fatal, that appeared to have been transmitted from the mother during birth. Perinatal prophylaxis, which has been recommended for infections due to group B streptococcus, should be evaluated for infections due to group A streptococcus.

Adult↗

Spread of serious disease-producing M3 clones of group A streptococcus among family members and health care workers.

Streptococcus pyogenes causes a variety of diseases ranging from mild pharyngitis to severe toxic shock syndrome (TSS) and acute rheumatic fever. Since 1987 there has been a resurgence of severe group A streptococcus infections including TSS, necrotizing fasciitis, and myositis. Using molecular and serotyping procedures, we recently studied two clusters of group A streptococcus disease that occurred within separate family units. The first cluster involved two family members (one with TSS and one with necrotizing fasciitis) and three health care workers who attended one of the index patients. The second cluster included a mother (with necrotizing fasciitis of the hand) and her three children. Group A streptococci isolated from individuals within both cluster groups were serotype M3;T3/13/B3264, and pulsed field gel electrophoresis revealed that all isolates except one had identical fingerprints of Sma I-digested chromosomal DNA. The findings demonstrate the potential for spread of serious group A streptococcus disease among individuals and the need for barrier protection when health care workers are exposed to secretions from infected individuals.

Adult↗

Vaginal colonization with group B streptococcus: a study in college women.

Vaginal specimens for culture of group B Streptococcus and anonymous questionnaires were obtained from 499 college women. Group B Streptococcus was isolated from 90 (18.0%) of the participants. A selective broth medium was more sensitive for detection of vaginal isolates (85 of 493; 17.2%) than was direct inoculation of blood agar plates (44 of 466; 9.4%). The most prevalent serotypes among the isolates were type III (37.9%) and type II (25.3%). Logit analysis identified four factors associated with a higher prevalence of vaginal colonization with group B Streptococcus. These organisms were isolated significantly more often from (1) women who had an intrauterine device (50% vs. 18.6%; P less than 0.001), (2) sexually experienced women (20% vs. 7.1%; P less than 0.02), (3) women studied during the first half of the menstrual cycle (26.5% vs. 14.5%; P less than 0.01), and (4) women 20 years of age or younger (21.4% vs. 14.8%; P less than 0.05). The prevalence of colonization with group B Streptococcus was not related to sexual practices, history of venereal disease, use of oral contraceptives, presence of gynecologic symptoms, use of antibiotics, race, educational level, marital status, or history of pregnancy.

Adult↗

Immunological investigation of infants with septicemia or meningitis due to group B Streptococcus.

Purified polysaccharide from type III group B Streptococcus contains both a type III-specific determinant and another determinant that is common to strains of serotypes other than type III. The polysaccharide contains sialic acid, galactose, heptose, glucose, glucosamine, and mannose. Serum antibody to this antigen was measured by means of a radioactive antigen-binding assay. Sera from 36 (67.9%) of 53 women with healthy newoborns contained antibody, a prevalence significantly different from that in sera from 15 women (13.3%) whose neonates developed septicemia or meningitis due to type III group B Streptococcus. Complete concordance for presence or absence of anticapsular antibody in sera from 14 women at delivery and in their neonates' cord sera was demonstrated; this concordance indicates transplacental transfer of antibody. Sera from each of four adults with invasive infection who were studied during convalescence contained antibody to the capsular polysaccharide of type III group B Streptococcus. In contrast, antibody was absent from 10 infants who had recovered from bacteremia, septicemia, and/or meningitis due to type III group B Streptococcus.

Antibodies, Bacterial↗

Streptococcus pluranimalium sp. nov., from cattle and other animals.

Strains from subclinical mastitis, from the genital tract and tonsils of cattle, from tonsils of a goat and a cat and from the crop and the respiratory tract of canaries were found to constitute a new streptococcal species, for which the name Streptococcus pluranimalium sp. nov. is proposed. Sequencing of 16S rRNA showed that Streptococcus thoraltensis and Streptococcus hyovaginalis were its closest known phylogenetic relatives. The new species showed some phenotypic resemblance to the poorly described species Streptococcus acidominimus, but whole-cell protein analysis and 16S rRNA sequencing revealed that the new species was only distantly related to the type strain of S. acidominimus. Identification of these bacteria, which showed heterogeneous biochemical reaction patterns, was most reliably made by whole-cell protein analysis. Nevertheless, a number of biochemical reactions can be used to differentiate S. pluranimalium from other animal streptococci. Strain LMG 14177T, isolated from mastitic milk of a dairy cow, was designated as the type strain of S. pluranimalium sp. nov.

Animals↗

The gene encoding IIAB(Man)L in Streptococcus salivarius is part of a tetracistronic operon encoding a phosphoenolpyruvate: mannose/glucose phosphotransferase system.

Glucose and mannose are transported in streptococci by the mannose-PTS (phosphoenolpyruvate:mannose phosphotransferase system), which consists of a cytoplasmic IIAB protein, called IIAB(Man), and an uncharacterized membrane permease. This paper reports the characterization of the man operon encoding the specific components of the mannose-PTS of Streptococcus salivarius. The man operon was composed of four genes, manL, manM, manN and manO. These genes were transcribed from a canonical promoter (Pman) into a 3.6 kb polycistronic mRNA that contained a 5'-UTR (untranslated region). The predicted manL gene product encoded a 35.5 kDa protein and contained the amino acid sequences of the IIA and IIB phosphorylation sites already determined from purified S. salivarius IIAB(Man)L. Expression of manL in Escherichia coli generated a 35 kDa protein that reacted with anti-IIAB(Man)L antibodies. The predicted ManM protein had an estimated size of 27.2 kDa. ManM had similarity with IIC domains of the mannose-EII family, but did not possess the signature proposed for mannose-IIC proteins from Gram-negative bacteria. From multiple alignment analyses of sequences available in current databases, the following modified IIC(Man) signature is proposed: GX3G[DNH]X3G[LIVM]2XG2[STL][LT][EQ]. The deduced product of manN was a hydrophobic protein with a predicted molecular mass of 33.4 kDa. The ManN protein contained an amino acid sequence similar to the signature sequence of the IID domains of the mannose-EII family. manO encoded a 13.7 kDa protein. This gene was also transcribed as a monocistronic mRNA from a promoter located in the manN-manO intergenic region. A search of current databases revealed the presence of IIAB(Man)L, ManM, ManN and ManO orthologues in Streptococcus mutans, Streptococcus pyogenes, Streptococcus pneumoniae and Enterococcus faecalis. This work has elucidated the molecular structure of the mannose PTS in streptococci and enterococci, and demonstrated the presence of a putative regulatory protein (ManO) within the man operon.

Amino Acid Sequence↗

Heterogeneity among 16S-23S rRNA intergenic spacers of species within the 'Streptococcus milleri group'.

The 16S-23S rRNA intergenic spacer has been suggested as a suitable region of the bacterial genome from which to derive useful taxonomic information, particularly with regard to identification at the species level. To investigate this approach as an aid to the identification of the three species comprising the 'Streptococcus milleri group' (SMG), the spacers of isolates of Streptococcus intermedius, Streptococcus anginosus and Streptococcus constellatus were amplified by PCR and length polymorphisms determined by agarose gel electrophoresis. Phenotypically atypical isolates which had been identified presumptively as belonging to these three species were also included. Spacers from two representatives of each spacer length found within the three SMG species were sequenced. 16S-23S rRNA intergenic spacer length polymorphisms allowed discrimination between S. anginosus (350 bp or 450 bp amplification product) and S. constellatus (380 bp amplification product), species that are difficult to differentiate phenotypically. S. intermedius (330 bp or 450 bp amplification product) and S. anginosus (350 bp or 450 bp amplification product) were not reliably distinguished by this method but are phenotypically distinct. Sequencing data demonstrated that the spacers had a central region of highly variable length flanked by conserved regions which included a single tRNA(Ala) gene. Polymorphism in the length of the 16S-23S spacer determined by PCR provides a rapid and useful adjunct to strain identification for S. anginosus and S. constellatus, which are not readily differentiated phenotypically.

Base Sequence↗

Streptococcus oligofermentans sp. nov., a novel oral isolate from caries-free humans.

Five streptococcal strains were isolated from dental plaque and saliva of caries-free humans. The cells were gram-positive, non-spore-forming, non-motile cocci that were arranged in short chains. The strains were catalase-negative, facultatively anaerobic and produced lactic acid exclusively from glucose fermentation. Biochemical analysis that used both conventional methods and the commercial API 20 Strep system showed that the five strains fermented only a few kinds of sugar. The mean DNA G + C content of the five novel strains was 39.5 +/- 0.8 mol%. Phylogenetic analysis based on 16S rDNA sequence homology indicated that the new isolates represented a novel member of the mitis group of the genus Streptococcus, related most closely to the recently described species Streptococcus sinensis. DNA-DNA relatedness between novel strain LMG 21535T and type strains of phylogenetically related species of oral streptococci was 7.1-16.4%. Therefore a novel Streptococcus species, Streptococcus oligofermentans sp. nov., is proposed. The type strain is LMG 21535T=AS 1.3089T.

Base Composition↗

Streptococcus halichoeri sp. nov., isolated from grey seals (Halichoerus grypus).

Phenotypic and phylogenetic studies were performed on six unidentified, Gram-positive, catalase-negative, chain-forming Streptococcus-like organisms recovered from grey seals. Biochemically the six strains were highly related to each other, but they did not appear to correspond to any recognized species of the genus Streptococcus. Comparative 16S rRNA gene sequencing studies confirmed that phylogenetically the strains were members of the genus Streptococcus, but sequence divergence values of greater than 3 % compared with reference streptococcal species demonstrated that the organisms from seals represent a novel species. SDS-PAGE analysis of whole-cell proteins confirmed the phenotypic distinctiveness of the seal organisms. Based on biochemical criteria and molecular chemical and genetic evidence, it is proposed that the unknown organism from seals be classified as a novel species, Streptococcus halichoeri sp. nov., the type strain of which is CCUG 48324T (= CIP 108195T).

Animals↗

Streptococcus equi subsp. ruminatorum subsp. nov., isolated from mastitis in small ruminants.

Six isolates of an unknown Gram-positive, catalase-negative, chain-forming, coccus-shaped organism isolated from ovine and caprine mastitis were characterized by phenotypic and molecular taxonomic methods. On the basis of cellular morphology and the results of biochemical tests, the organism was tentatively identified as a streptococcal species. Comparative 16S rRNA gene sequencing studies confirmed that the organism is a member of the genus Streptococcus, with Streptococcus equi as its closest phylogenetic relative (98.8 % similarity). DNA-DNA pairing studies showed that the unidentified organism displayed more than 70 % relatedness to the type strains of S. equi subsp. equi and subsp. zooepidemicus. Despite the relatively high DNA-DNA reassociation values, biotyping and ribotyping allowed clear differentiation of the unknown bacterium from the two recognized subspecies of S. equi. On the basis of phenotypic and molecular genetic evidence, it is proposed that the unknown Streptococcus isolates from ovine and caprine mastitis be classified as a novel subspecies, Streptococcus equi subsp. ruminatorum subsp. nov. The type strain is CECT 5772(T) (=CCUG 47520(T)=Mt 167(T)).

Animals↗

Streptococcus marimammalium sp. nov., isolated from seals.

Two strains of an unidentified, Gram-positive, catalase-negative, chain-forming, coccus-shaped organism recovered from seals were characterized using phenotypic and molecular taxonomic methods. Based on morphological and biochemical criteria the strains were tentatively identified as streptococci but they did not appear to correspond to any recognized species of the genus Streptococcus. Comparative 16S rRNA gene sequencing studies showed that the strains were closely related to each other and confirmed their placement in the genus Streptococcus. Sequence divergence values of >5 % with reference streptococcal species demonstrated the organisms from seals represent a novel species. SDS-PAGE analysis of whole-cell proteins confirmed that the two organisms were closely related to each other but were different from all currently defined streptococcal species. Based on biochemical criteria, molecular chemical and molecular genetic evidence, it is proposed that the unknown isolates from seals be assigned to a novel species of the genus Streptococcus, Streptococcus marimammalium sp. nov. The type strain is M54/01/1T (=CCUG 48494T=CIP 108309T).

Animals↗

16S-23S rRNA intergenic spacer region sequence variation in Streptococcus thermophilus and related dairy streptococci and development of a multiplex ITS-SSCP analysis for their identification.

The 16S-23S rRNA internal transcribed spacer (ITS) region of several Streptococcus thermophilus strains and some related dairy streptococci, S. macedonicus, S. salivarius and S. bovis, was analysed by sequence analysis. All the Streptococcus species were easily discriminated on the basis of sequence variations principally located upstream and downstream of the region encompassing the double-stranded processing sites and the tRNA(Ala) gene. Comparison between tRNA(Ala) gene sequences highlighted a high level of sequence conservation among the Streptococcus species investigated despite their belonging to separated phylogenetic clusters, i.e. the S. salivarius and S. bovis rRNA groups. A low but significant degree of variability was detected among the S. thermophilus strains, allowing the identification of four different ITS sequences. Similarity analysis of the ITS sequences showed that the Streptococcus species were clustered in two main branches, one containing S. macedonicus and S. bovis strains, and one containing S. thermophilus and S. salivarius strains. With the aim of developing a rapid tool for the identification of the dairy streptococci species a multiplex ITS-SSCP analysis of two discrete regions within the ITS locus was carried out.

Bacterial Typing Techniques↗

Abscess forming ability of streptococcus milleri group: synergistic effect with Fusobacterium nucleatum.

The abscess forming abilities of "Streptococcus milleri" strains (Streptococcus constellatus, Streptococcus anginiosus, and Streptococcus intermedius) isolated from dentoalveolar abscesses and the synergistic effect of Fusobacterium nucleatum co-inoculated with the isolates were examined on a mouse subcutaneous abscess model. Five days after inoculation, all S. milleri strains formed abscesses, which showed less pathological spread to surrounding connective tissues than those formed by Staphylococcus aureus 209P strain and were similar to those by F. nucleatum ATCC25586. When each S. milleri strain and F. nucleatum were co-inoculated, abscess sizes and each bacterial number recovered from abscesses increased in comparison to those treated by bacterial mono-inoculation of each S. milleri strain or F. nucleatum alone. The strongest synergistic effect was observed in the combination of S. constellatus and F. nucleatum. In a time course experiment with this combination, the recovery of S. constellatus subsequently decreased after the decrement of F. nucleatum, and it appeared that the association with F. nucleatum maintained the bacterial number of S. constellatus in the abscess. The cell-free supernatant of F. nucleatum had a tendency to increase the abscess size caused by S. constellatus in this model. When S. constellatus was cultured with F. nucleatum culture supernatant in vitro, growth enhancement in the early phase was observed. Furthermore, the phagocytic killing of S. constellatus by human polymorphonuclear leukocytes (PMNs) was significantly suppressed and the PMN membranes appeared to be injured by addition of the F. nucleatum culture supernatant. These results suggest that the pathogenicity of S. milleri strains in odontogenic infections may be enhanced by the co-existence of F. nucleatum.

Abscess↗

Molecular characterization of dextranase from Streptococcus rattus.

The complete nucleotide sequence of the dextranase gene of Streptococcus rattus ATCC19645 was determined. An open reading frame of the dextranase gene was 2,760 bp long and encoded a dextranase protein consisting of 920 amino acids with a molecular weight of 100,163 Da and an isoelectric point of 4.67. The S. rattus dextranase purified from recombinant Escherichia coli cells showed dextran-hydrolyzing activity with optimal pH (5.0) and temperature (40 C) similar to those of dextranases from Streptococcus mutans and Streptococcus sobrinus. The deduced amino acid sequence of the S. rattus dextranase revealed that the dextranase molecule consists of two variable regions and a conserved region. The variable regions contained an N-terminal signal peptide and a C-terminal cell wall sorting signal; the conserved region contained two functional domains, catalytic and dextran-binding sites. This structural feature of the S. rattus dextranase is quite similar to that of other cariogenic species such as S. mutans, S. sobrinus, and Streptococcus downei.

Amino Acid Sequence↗

Differentiation of human and animal strains of Streptococcus dysgalactiae by pulsed-field gel electrophoresis.

The genetic diversity among 54 human isolates and 33 animal isolates belonging to the species Streptococcus dysgalactiae (20 alpha-haemolytic Streptococcus dysgalactiae, 23 Streptococcus equisimilis, 43 group G streptococci and one group L streptococcus) was evaluated by macrorestriction analysis of chromosomal DNA with SmaI and resolution by pulsed-field gel electrophoresis. This technique revealed a high degree of intraspecies polymorphism, leading to the differentiation of 80 distinct banding patterns, and identified the presence of two major clusters, one containing isolates of human origin and the other isolates of animal origin. These results suggest than human and animal isolates of S.dysgalactiae are genetically distinct, and support the recent proposal of the subspecies S. dysgalactiae subsp. equisimilis for human isolates. The heterogeneity revealed within isolates from the same host type indicates that pulsed-field gel electrophoresis is a powerful epidemiological tool for studying S. dysgalactiae infections.

Animals↗

[Relationship of serum streptococcal antibodies to carrier state of beta-hemolytic streptococcus in throats of healthy schoolchildren].

The relationship of ASO, ASK and ASP to carrier state of beta-hemolytic streptococcus in throats of healthy schoolchildren was examined from September, 1980 to May, 1987. The responses to ASO significantly increased in children who detected group A, C and G streptococcus, and ASK in group A and G, and ASP in group A. The responses were especially high in children who detected group A, C and G streptococcus one after the other through the term of research, and in children detected another T type of streptococcus one after the other. Relatively high titers of ASO and ASK were obtained in sera of children from whom some groups or T types of streptococci were isolated several detection times. ASP titers were not concerned with these times. The elevation of antibodies in paired samples were found as follows; 7 cases in ASO, ASK and ASP together, 42 cases in ASO and ASK together, 3 cases in ASK and ASP together, 2 cases in ASO and ASP together, 41 cases ASO only, 38 cases in ASK only, 7 cases in ASP only. Among these cases with elevation of antibodies titers, several kinds of the change of streptococcal carrier state were observed between obtaining serum of twice. Especially 15 cases with change of carrier state from group A to group A were noticed. Among these 15 cases, 11 cases isolated another T type strain through the term of research, or found to contain the research-time when no streptococcal strain was detected. Both long term carrier state of the same group or the same type of streptococcal strain and change of carrier state of streptococcal group or T type lead the elevation of ASO and ASK titers.

Antibodies, Bacterial↗