Lancefield Group Research Articles

Molecular characterization of clinical isolates of M non-typable group A streptococci from invasive disease cases

Currently there are 93 validated M serotypes of Streptococcus pyogenes, Lancefield group A streptococcus (GAS), and >130 emm genotypes. A marked increase in the number of non-typable GAS isolates (2 % in 2000, 4 % in 2001 and 9 % in 2002) from invasive disease cases referred to the authors' reference laboratory was noted during 2000-2002. A total of 217 (92 %) were from blood cultures, 14 (6 %) from deep abscesses and five (2 %) from aspirates. The clinical manifestations included bacteraemia, septicaemia, cellulitis, meningitis, necrotizing fasciitis and toxic-shock syndrome. In order to establish whether this increase was due to the emergence of novel types or the unavailability of M-typing sera, these isolates were subjected to emm sequencing. A total of 144 isolates (61 %) belonged to M types for which sera were no longer available; 112 (48 %) belonged to higher M types, including emm83.1 (9 %), emm94 (8 %) emm87 (6 %) and emm89 (6 %); and 32 (13 %) belonged to lower M types that were not commonly isolated in the UK, and included M25, M43, M49, M64, M73 and M74. Sixty-six (28 %) of the isolates belonged to newly designated emm types. Other isolates belonged to the novel emm types st2147, STNS1033 and st854, recently registered in the Centers for Disease Control (CDC) database by other laboratories. One novel emm type, st2161, was isolated from an injecting drug user. There were differences in the type distribution of these isolates according to geographic location. However, 90 % of emm93, one of seven predominant emm types identified amongst the collection of M non-typable (MNT) isolates, were isolated from the London region.

Acute peritonitis and salpingitis associated with streptococcal toxic shock syndrome caused by Lancefield group G α-haemolytic Streptococcus dysgalactiae subsp. equisimilis

The authors treated a patient for what appears to be the first reported occurrence of acute peritonitis and salpingitis associated with streptococcal toxic shock syndrome (STSS). This was caused by Lancefield group G alpha-haemolytic Streptococcus dysgalactiae subsp. equisimilis TKCH2004-001. The isolate showed M protein type stc36 and carried the spegg gene. To the best of the authors' knowledge, the present report represents the first case of STSS complicating acute peritonitis and salpingitis caused by Lancefield group G alpha-haemolytic S. dysgalactiae subsp. equisimilis.

Speciation of clinical isolates of group C haemolytic streptococci harvested in North Yorkshire, England

All Lancefield group C haemolytic streptococci isolated from routine clinical specimens over four years were collected to determine the species composition. Of a total of 102 viable isolates studied, 71 (70%) were large colony-forming Streptococcus dysgalactiae equisimilis; they were from patients of all ages (predominantly the 11–20 year age group), mainly (72%) from the throat or wound swabs but some from genital, blood and other sites. Thirty (29%) of the isolates were of the small colony-forming ‘ Streptococcus milleri’ group ( Streptococcus anginosus = 21, Streptococcus constellatus = 9) from patients in the age group of 10–70 years (predominantly 21–30 years); two-thirds of these isolates were from the throat, with others from genital, wound and other sites. Comparison of results with studies of earlier collections of group C streptococci showed broad similarity, although other researchers have often found a higher proportion of S. milleri strains. The continuing low incidence of human infection with Streptococcus zooepidemicus here is confirmed.

Genetic and Biochemical Properties of Streptococcal NAD-glycohydrolase Inhibitor

The gene encoding streptolysin O (slo), a cytolysin of hemolytic streptococci, is transcribed polycistronically from the promoter of the preceding NAD-glycohydrolase (NADase) gene (nga). Between nga and slo, a putative open reading frame (orf1) is located whose function has been totally unknown. Present investigation demonstrated that the orf1 encodes a protein designated as streptococcal NADase inhibitor (SNI). From its nucleotide sequence, SNI was inferred to comprise 161 amino acid residues and the deduced molecular weight was 18,800. This protein was detectable only within cells. Coexpression of SNI was essential for production of streptococcal NADase, and NADase precursor existed as an inactive complex with SNI, in recombinant Escherichia coli. Monomeric NADase and SNI rapidly formed in vitro a stable heterodimer complex in the ratio 1:1, resulting in complete suppression of the hydrolase activity. Unlike other bacterial NADase inhibitors, SNI was thermostable. This protein, coexpressed and complexed with NADase, may protect the producer cocci from exhaustion of NAD.

Characterization of fluoroquinolone-resistant β-hemolytic Streptococcus spp. isolated in North America and Europe including the first report of fluoroquinolone-resistant Streptococcus dysgalactiae subspecies equisimilis: Report from the SENTRY Antimicrobial Surveillance Program (1997–2004)

β-Hemolytic streptococci are common bacterial pathogens that can cause serious invasive disease, and although this group of species has remained susceptible to the fluoroquinolone class, resistant strains have been reported. This multicenter investigation determined the rate of fluoroquinolone-resistant β-hemolytic streptococci using the SENTRY Antimicrobial Surveillance Program network data (1997–2004). Forty-seven surveillance culture isolates of β-hemolytic streptococci from North America and Europe with elevated levofloxacin MIC results (2 to >32) μg/mL were tested for susceptibility to other fluoroquinolones including ciprofloxacin, garenoxacin, gatifloxacin, gemifloxacin, and moxifloxacin using reference broth microdilution and Etest (BIODISK, Solna, Sweden) methods. Strains were characterized using polymerase chain reaction and sequencing to detect mutations in the quinolone-resistance determining region (QRDR). The β-hemolytic streptococci isolates with reduced fluoroquinolone susceptibility included the following Lancefield groups: A ( Streptococcus pyogenes; 9 strains), B ( Streptococcus agalactiae; 24 strains), C and G (14 strains). Vitek and API 20 strep (bioMerieux, Hazelwood, MO) identification systems, as well as conventional biochemical methods and colony morphology, were used to confirm the organism identifications. The overall potency (MIC 90 in μg/mL) for the fluoroquinolones against all tested β-hemolytic streptococci showed the following rank order: gemifloxacin (0.06) > garenoxacin (0.12) > moxifloxacin (0.25) > gatifloxacin (0.5) > levofloxacin = ciprofloxacin (1). The rate of levofloxacin-resistant β-hemolytic streptococci in the SENTRY program was 0.14% (Europe) and 0.51% (North America) during the study period. All levofloxacin-resistant strains tested by molecular methods had significant mutations in either parC (position 79 or 83) and/or gyrA (position 81 or 85). All but 2 isolates with high-level resistance to levofloxacin (>32 μg/mL) had gyrA mutations. Strains with lower MIC values to levofloxacin (2–4 μg/mL) only had mutations in parC. The increasing rate of fluoroquinolone-resistant streptococci including Streptococcus pneumoniae, viridans group streptococci, and the more recently reported β-hemolytic streptococci, is becoming a clinical concern due to the morbidity and mortality caused by these pathogens.

Clonal Relationships between Invasive and Noninvasive Lancefield Group C and G Streptococci and emm -Specific Differences in Invasiveness

Lancefield group G and group C streptococci (GGS and GCS, respectively) are pathogens responsible for a number of life-threatening infections. A collection of 116 recent (1998 to 2004) invasive (n = 28) and noninvasive (n = 88) GGS and GCS clinical isolates from Portugal were characterized. All isolates were identified as Streptococcus dysgalactiae subsp. equisimilis and characterized by emm typing and DNA macrorestriction profiling using pulsed-field gel electrophoresis (PFGE). emm typing revealed the presence of 22 distinct types, including 3 novel types. PFGE identified 14 clones with more than two isolates, but over half of the isolates were concentrated in 3 large clones. Individual clones and emm types showed a low level of association, since the majority of the clones included more than one emm type and the same emm type was found among diverse genetic backgrounds. Two emm types, stg2078 and stg10, were significantly more frequent among invasive isolates, and another two, stg6792 and stg166b, were present only in noninvasive isolates, suggesting a correlation between emm type and invasive disease potential.

Severe Streptococcus infection in spotted hyenas in the Ngorongoro Crater, Tanzania

In a population of spotted hyenas ( Crocuta crocuta) monitored between 1996 and 2005 in the Ngorongoro Crater, Tanzania, 16 individuals from five of eight social groups displayed clinical signs of an infection, including severe unilateral swelling of the head followed by abscess formation at the mandibular angle, respiratory distress, mild ataxia, and lethargy. Two (12.5%) of these 16 individuals died within days of developing signs. Clinical signs in hyenas were first noted in 2001, and most cases occurred between September 2002 and February 2003, suggesting an outbreak of infection during this period. Histopathological examination of internal organs from one hyena that died with signs revealed morphological changes consistent with severe bacterial infection. Phenotypic examination and phylogenetic analysis of the 16S rRNA gene of the causative agent of infection revealed a Lancefield group C Streptococcus with a high level of homology to S. equi subsp. ruminatorum, a subspecies of S. equi recently described in domestic sheep ( Ovis aries) and goats ( Capra hircus) with mastitis in Spain. Strains similar to this bacterium were also isolated from two hyenas without obvious clinical signs, suggesting that hyenas may be ‘carriers’ of this bacterium, and from a sympatric Burchell's zebra ( Equus burchelli), a herbivore species often consumed by hyenas. To our knowledge this is the first report of a Streptococcus infection in these two wildlife species. The high genetic similarity between the hyena and zebra isolates indicates that inter-specific transmission may occur, possibly when hyenas consume infected zebra carcasses.

Molecular epidemiology of macrolide resistance in β-haemolytic streptococci of Lancefield groups A, B, C and G and evidence for a new mef element in group G streptococci that carries allelic variants of mef and msr(D)

To study the molecular mechanisms of erythromycin resistance in beta-haemolytic streptococci of Lancefield groups A, B, C and G. Erythromycin-resistant clinical isolates from North East Scotland were collected over 2 years. Resistance phenotypes were determined by disc diffusion and MICs by Etest. Resistance genes mef, msr(D), erm(B) and erm(TR) were identified by PCR and mef and msr(D) were sequenced. Erythromycin resistance prevalence was 1.9% in group A streptococci (31 of 1625), 4.3% in group B (53 of 1233), 3.8% in group C (18 of 479) and 6.2% in group G (64 of 1034). The numbers of resistant isolates available were 26, 42, 9 and 52 in each group respectively. The majority of resistant isolates in groups A (57.7%, 15 of 26), B (88.1%, 37 of 42) and G (90.4%, 47 of 52) were MLS(B). The contribution of M phenotype was significant in groups C (77.8%, 7 of 9) and A (42.3%, 11 of 26). Group A isolates carried mef(A) and group B carried mef(E) exclusively. A mef sequence distinct from mef(A) and mef(E) was identified in group G and was associated with a new msr(D) sequence. These sequence variants appear to be part of a new genetic element that is inserted in the comEC gene. A bimodal distribution of erythromycin MICs was noted in erm(TR) isolates. The results demonstrate significant differences in the mechanisms of macrolide resistance amongst different Lancefield groups in the same geographical area. New sequences show that resistance mechanisms are still evolving.

Evaluation of a nested PCR test and bacterial culture of swabs from the nasal passages and from abscesses in relation to diagnosis of Streptococcus equi infection (strangles)

Streptococcus equi is the cause of strangles in horses. To improve diagnostic sensitivity, development and evaluation of DNA-based methods are necessary. To evaluate diagnostic methods and observe the pattern of bacterial shedding during natural outbreaks. Two herds with natural outbreaks of strangles were visited over a period of 15 weeks and 323 samples originating from 35 horses investigated. The diagnostic use of a nested PCR test was evaluated using a collection of 165 isolates of Lancefield group C streptococci (species specificity) and swabs from nasal passages or from abscesses from horses infected with S. equi (diagnostic sensitivity). All 45 S. equi isolates tested positive in the nested PCR, whereas no amplicon was formed when testing the other 120 Lancefield group C isolates. A total of 43 samples were collected from 11 horses showing clinical signs of strangles during the study period. The diagnostic sensitivity for PCR test was 45% and 80% for samples from the nasal passages and abscesses, respectively; the corresponding diagnostic sensitivity for cultivation was 18% and 20%. The diagnostic sensitivity was significantly higher for PCR than for bacterial cultivation. Furthermore, the shedding of S. equi in 2 infected horse populations was evaluated. An intermittent shedding period of S. equi of up to 15 weeks was recorded in this part of the study. It was also shown that shedding of S. equi occurred both from horses with and without clinical signs. The nested PCR test represents a species-specific and -sensitive method for diagnosis of S. equi from clinical samples. It may, however, be desirable in future to develop detection methods with high diagnostic sensitivity and specificity without the potential problems inherent in nested PCR.

<I>Streptococcus dysgalactiae</I> subsp. <I>equisimilis</I>の遺伝子解析による<I>emm</I>型別と経口抗菌薬感受性

A total of 593 beta-hemolytic streptococci belonging to Lancefield group A (GAS), group C (GCS) or group G (GGS) according to agglutination tests were collected from 11 medical institutions between September 2003 and October 2005. In total, 128 strains were identified as Streptococcus dysgalactiae subsp. equisimilis (S. equisimilis) using physiological tests. Of these strains, 5 strains were agglutinated to Lancefield group A, 17 strains to group C, and 106 strains to group G. Most of these strains were largely isolated from clinical specimens collected from young patients with respiratory infections and middle-aged patients (in their 40s); most of the strains were isolated from blood, atretic pus, or joint fluid. Genetic analysis of the emm gene encoding the M protein revealed that these strains could be classified into 27 types. Also, many emm types were found in strains isolated from normally aseptic clinical specimens. In addition, all strains tested had slo, sagA, and skcg genes, which contributed to their virulence. The susceptibility of the strains to oral penicillin and cephalosporin antibiotics was excellent, with MICs ranging from 0.016 to 0.031mg/mL. In contrast, strains carrying the macrolide resistant elements of the ermA, ermB, and mefA genes and strains showing a high resistance to levofloxacin were also confirmed in this study. These results suggest that beta-hemolytic streptococci, except for S. pyogenes and S. agalactiae, should be reconsidered as a causative pathogen in streptococcal infections.

Isolation and characterization of Streptococcus spp. group B in Nile tilapias (Oreochromis niloticus) reared in hapas nets and earth nurseries in the northern region of Parana State, Brazil

The objective of this study was to isolate and characterize Streptococcus spp. in Nile tilapias (Oreochromis niloticus) reared in net-pens and earth nurseries. Eight intensive tilapia-rearing farms were investigated in north Paraná, Brazil from April 1st 2001 to April 30th 2002. The fish were reared in a system of hapas nets on four farms and in earth nurseries on other four farms. A total of 370 samples were analyzed of material collected from 120 fish (brain, liver, kidney, skin scrapes, ascites liquid and eye) that were sown on BHI agar (Brain Heart Infusion) supplemented with 1% yeast extract and sheep blood. Streptococcus spp. was isolated in 36 of the samples (18 brain, eight liver, eight kidney and two ascites liquid) from 25 fish. Streptococci were isolated in both systems, almost in the same proportion. First the streptococci were characterized by the catalase and esculin test, growth in methylene blue and sodium chloride at 6.5%. They were classified in groups by the Slidex Strepto-Kit (BioMerieux, France). The phenotypic characteristics were determined by the Api 20 Strep microtest system (BioMerieux, France). The 36 Streptococcus spp. samples did not present hemolysis and were classified as Lancefield group B. Further 16 samples were identified as Streptococcus agalactiae and 20 were not identified by the Api 20 Strep, but presented the same biochemical profile described for the reference strain of Streptococcus difficile (ND-2-22).

Severe acute rhabdomyolysis associated with Streptococcus equi infection in four horses

Four Quarter Horses (9 months to 7 years of age) with submandibular lymphadenopathy and firm muscles (palpation of which elicited signs of pain) were evaluated; in general, the horses had a stiff gait, and 3 horses became recumbent. Streptococcus equi was cultured from aspirates of lymph nodes or samples of purulent material collected from the auditory tube diverticula. Once the horses were recumbent, their condition deteriorated rapidly despite aggressive antimicrobial and antiinflammatory treatment, necessitating euthanasia within 24 to 48 hours. One horse did not become recumbent and recovered completely. Among the 4 horses, common clinicopathologic findings included neutrophilia, hyperfibrinogenemia, and high serum activities of creatine kinase and aspartate aminotransferase. Necropsies of the 3 euthanatized horses revealed large, pale areas most prominent in the semimembranosus, semitendinosus, sublumbar, and gluteal muscles that were characterized histologically by severe acute myonecrosis and macrophage infiltration of necrotic myofibers. Streptococcus equi was identified in sections of affected muscle by use of immunofluorescent stains for Lancefield group C carbohydrate and S. equi M protein. In the 4 horses of this report, acute severe rhabdomyolysis without clinical evidence of muscle atrophy or infarction was associated with S. equi infection; rhabdomyolysis was attributed to either an inflammatory cascade resembling streptococcal toxic shock or potentially direct toxic effects of S. equi within muscle tissue.

Molecular Diagnosis of Necrotizing Fasciitis by 16S rRNA Gene Sequencing and Superantigen Gene Detection

We report the use of molecular techniques in the diagnosis of a case of culture-negative necrotizing fasciitis occurring in a 32-year-old female with no significant past medical history and who died within 36 hours of admission. Paraffin-embedded tissue sections from the popliteal fossa region obtained at autopsy showed hemorrhage, necrosis, and mild inflammation by hematoxylin and eosin staining. Tissue gram stain showed numerous gram-positive organisms arranged in clusters. The sequences of the first 500 bp of bacterial 16S rRNA gene amplified from the lesion were identical to a Lancefield group A beta-hemolytic Streptococcus pyogenes. Streptococcal pyrogenic exotoxin A and B superantigen genes were detected and an emm type 1 was determined by polymerase chain reaction and sequencing from the lesion. This confirmed the etiology of the patient's rapid deterioration with multisystem organ failure.

Clinical, phenotypic, and genotypic evidence for Streptococcus sinensis as the common ancestor of anginosus and mitis groups of streptococci

In 2002, we reported the discovery of a novel species of viridans streptococcus, Streptococcus sinensis. Recently, we reported the isolation of two more strains of S. sinensis. Clinically, S. sinensis is a definite cause of infective endocarditis, a characteristic mainly pertaining to the mitis group of streptococci. Phenotypically, two of the three S. sinensis isolates were Lancefield group F, a characteristic of the anginosus group. However, none of the three strains possess the caramel smell typical of this group of streptococci. Biochemically, S. sinensis was identified in 56% of the time as members of the anginosus group, and in 33% of the time as members of the mitis group. These clinical and phenotypic properties should be governed by the presence/absence or expressivity of particular genes in the S. sinensis genome. Genotypically, phylogenetic analysis using 16S rRNA gene sequences showed that S. sinensis branched out as the first branch in the anginosus group, implying that it is the ancestor of the other members of this group. However, the bootstrap value for S. sinensis clustered with members of the anginosus group is only 47%, meaning that it is often not clustered with members of this group, but the mitis group. Furthermore, the differences in the 16S rRNA gene sequences between S. sinensis and Streptococcus intermedius (3.7%) and those between S. sinensis and Streptococcus gordonii (3.6%) are almost the same. All these indicated that it is very likely that S. sinensis is the common ancestor of the anginosus and mitis groups of streptococci. Complete genome sequencing of S. sinensis and comparative genomics studies on the S. sinensis genome and genomes of members in the anginosus and mitis groups should reveal clues to the underlying genotypic differences that govern the different phenotypic properties of the two groups of streptococci, such as why streptococci of the anginosus group are prone to cause abscess formation but not infective endocarditis as compared to other viridans streptococci.

Streptococcus anginosus group: Clinical significance of an important group of pathogens

The Streptococcus anginosus group was first discovered in the 1950s and over time has evolved to consist of three species designated S. anginosus, S. constellatus , and S. intermedius . Investigation of these bacteria over the last few decades has revealed that infections caused by these gram-positive cocci have significant variability in their clinical presentations. In addition, studies have shown that hemolysis, Lancefield group reaction, and other laboratory techniques commonly used to identify bacteria are inconsistent throughout the S. anginosus group. Clinical review has shown that while the S. anginosus group is found as normal flora in humans, these bacteria are pathogens associated with infection in multiple body sites. In addition, the S. anginosus group has been shown to be strongly associated with abscess formation, and recent studies have attempted to investigate the pathogenic mechanisms of these bacteria. While little antibiotic resistance has been noted, treatment of abscesses often requires surgical drainage.

Balanitis caused by <I>Streptococcus pyogenes</I>: a report of two cases

Streptococcus pyogenes (the Lancefield group A streptococcus) is a cause of pharyngitis and impetigo. However, it has rarely been implicated as a sexually transmitted pathogen. We herein report two cases of severe balanitis due to S. pyogenes in sexually active men. It is postulated that penile cellulitis developed following the invasion of S. pyogenes through a traumatic abrasion acquired during fellatio performed by commercial sex workers. Both patients were treated successfully with oral administration of penicillin.

Development of a real-time PCR assay on the Roche Light-Cycler for the detection of erm and mef erythromycin resistance genes in β-haemolytic streptococci

Sir, Although erythromycin resistance in streptococci is predominantly due to target site modification by rRNA methylases (encoded by erm genes) or drug efflux (encoded by mef genes), novel resistance mechanisms continue to emerge. Furthermore, susceptibility to macrolides, lincosamides, streptogramins and ketolides varies depending on the distribution of different erythromycin resistance mechanisms. Therefore, phenotypic and genotypic erythromycin resistance surveillance data are required as an adjunct in determining therapy for streptococcal infections. In the UK, erythromycin resistance in Lancefield group B (GBS), C (GCS) and G (GGS) streptococci is increasing, but data on the molecular basis of resistance are limited. PCR with agarose gel analysis or microwell-format probe hybridization can be used to detect erm and mef genes; however, both require additional handling steps and gel analysis lacks the specificity of sequencing or hybridization analyses. Real-time PCR has the dual benefits of rapid amplification and specific detection, occurring simultaneously in the same tube, reducing the risks of contamination. The aim of this study was to develop a real-time PCR assay to detect both erm and mef genes and characterize erythromycin resistance genes in GBS, GCS and GGS isolated from clinical samples. This approach has not been used before in detecting erm and/or mef genes in streptococci. Positive control strains containing erm and mef genes used were: 02C1064 [Streptococcus pyogenes, mef(A/E)]; 02C1110 [S. pyogenes, erm(A) subclass erm(TR)]; RN1389 [Staphylococcus aureus, erm(A)]; JH2-2 [Enterococcus faecalis, erm(B)] and RN4220 [S. aureus, erm(C)]. Sixty-one erythromycinresistant streptococci (14 GBS, four GCS and 43 GGS) collected from patients with community-acquired infections at two different hospitals during two different 6 month periods were also tested.

The prevention of neonatal group B streptococcal disease: a report by a working group of the Medical Screening Society

Streptococcus agalactiae, or Lancefield group B streptococcus (GBS), is the most frequent cause of serious bacterial sepsis, including neonatal meningitis, in UK neonates. Early-onset neonatal GBS infection, but not late-onset, can be prevented by screening to identify high-risk pregnancies and administering penicillin during delivery. A vaccine has been developed as an alternative means of prevention but it is awaiting a randomized trial before being available for general use. In this review we examine the published literature to assess the morbidity and mortality attributable to neonatal GBS infection, quantify the screening performance of the two alternative modes of screening (microbiological and risk factor based), review the evidence on the efficacy of the vaccine, and estimate the numbers of deaths and cases of serious disability that each strategy in turn might prevent in the UK, in order to assess the most effective means of prevention for the UK.

Streptococcus castoreus sp. nov., isolated from a beaver (Castor fiber)

A previously undescribed, Gram-positive, catalase-negative, Streptococcus-like organism originating from a European beaver (Castor fiber) was subjected to a taxonomic study. The organism displayed beta-haemolytic activity and gave a positive reaction with Lancefield group A antisera. Based on the results of biochemical testing, the organism was tentatively identified as a member of the genus Streptococcus, but it did not correspond phenotypically to any recognized species of this genus. Comparative 16S rRNA gene sequencing studies confirmed this assignment, with the bacterium forming a hitherto unknown subline within the genus. Sequence divergence values of greater than 3 % from other reference streptococcal species, however, demonstrated that the unidentified coccus-shaped organism represents a hitherto unknown species. Based on phenotypic and molecular phylogenetic evidence, it is therefore proposed that the unknown organism from a beaver be classified as a novel species, Streptococcus castoreus sp. nov. The type strain is M605815/03/2(T) (=CCUG 48115(T)=CIP 108205(T)).

Biochemical analysis, cpn60 and 16S rDNA sequence data indicate that Streptococcus suis serotypes 32 and 34, isolated from pigs, are Streptococcus orisratti

Streptococcus suis serotypes have traditionally been identified by morphology, biochemical profiling and serotyping. Analysis of the sequences of 16S rRNA and cpn60 genes of the 35 characterized serotypes of S. suis led to the observation that two serotypes 32 and 34, are significantly distinct from other S. suis serotypes and may represent a distinct species. Here we present DNA sequence data and biochemical profiles which indicate that S. suis serotypes 32 and 34, isolated from pigs, are clustered with Streptococcus orisratti, a Voges-Proskauer negative, alpha-haemolytic, aesculin-hydrolytic, Lancefield group A streptococcus isolated from the teeth of rats.