INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY, Apr. 1987, p. 160-162
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Copyright 0 1987, International Union of Microbiological Societies
Streptococcus suis sp. nov., nom. rev.
RENATE KILPPER-BALZ AND KARL HEINZ SCHLEIFER”
Lehrstuhl fur Mikrobiologie, Technische Universitat Miinchen, Arcisstrasse 21, 0-8000Munich 2, Federal
Republic of Germany
Chemotaxonomic and deoxyribonucleic acid homology studies were carried out on various ‘‘Streptococcus
suis” strains to clarify their taxonomic position. The results of the present study indicate that these strains
belong to one species, namely, S. suis (ex Elliott) nom. rev. Strain NCTC 10234 is proposed as the type strain.
“Streptococcus suis” is an important pathogen in pigs. It
was originally isolated from cases of bacteremia and menin-
gitis in piglets (2, 4, 15). More recently, however, “ S . suis”
has been isolated from pigs with respiratory diseases (5, 11).
The name “S. suis” has never been formally proposed or
described and is, therefore, not on the Approved Lists of
Bacterial Names (14).
Originally described strains were found to be serologically
groupable into Lancefield groups R, S, RS, and T, and some
were nongroupable (3). Other authors (11, 15) described
eight different serovars of “ S . suis.” All these strains are
mlorphologicallyand biochemically very similar. In the pres-
ent study, strains of the different Lancefield groups and
serovars were investigated by deoxyribonucleic acid (DNA)
hybridization and cell wall analyses to determine genetic
relationships of these serologically heterogeneous organisms
anid to clarify their taxonomic status.
MATERIALS AND METHODS
The strains used in this study are shown in Table 1. They
were cultivated in a medium consisting of (per liter of
distilled water) 10 g of casein hydrolysate, 10 g of yeast
extract, 5 g of D-glucose, 5 g of sodium chloride, 3 g of
sodium acetate, 0.5 g of L-cysteine hydrochloride, and 1 ml
of Tween 80 at pH 7.2 under microaerophilic conditions at
35°C for 18 h. For the isolation of DNA, cells were harvested
at the end of the exponential growth phase after 2 h of
incubation with penicillin according to the method of Kilp-
per-Balz and Schleifer (7). Methods for the preparation of
filter-bound and radioactive DNA as well as optimal hybrid-
ization conditions and determination of guanine-plus-
cytosine (G+C) contents by thermal denaturation have been
described previously (6, 9). Isolation of cell walls and
determination of peptidoglycan types and cell wall sugar
compositions were done with cells in the stationary phase, as
previously described (12, 13).
RESULTS AND DISCUSSION
The results of DNA-DNA hybridization studies showed
thcat all “S. suis” strains investigated are highly related,
regardless of their Lancefield group or serovar. DNA relat-
* Corresponding author.
160
Vol. 37, No.
edness among different strains averaged more than 80%
(Table 2), confirming their relationship at the species level.
Cell wall composition and values of moles percent G+C
have been summarized in Table 1. Most strains of “ S . suis”
studied contained the lysine-direct peptidoglycan type (di-
rectly cross-linked stem peptides containing lysine as the
diamino acid [13]). This peptidoglycan type was found
among streptococci only in strains of “ S . suis” and S . oralis
(8). However, in contrast to S . oralis, strains of “S. suis”
contained rhamnose and lacked ribitol as major cell wall
polysaccharide constituents. In two strains of “ S . suis”,
namely, strain Henrichsen 11538 (serovar S) and strain
Henrichsen 2651 (Lancefield group RS), at least part of the
stem peptides of their peptidoglycan was not directly cross-
linked but linked by an interpeptide bridge consisting of
alanine (ly~ine-[alanine]~-~; Table 1).These two strains were
genetically closely related to typical strains of “S. suis”
(Table 2).
Because of the reaction with group D antiserum, it had
been suggested that “S. suis” belongs to group D strepto-
cocci together with “ S . bovis” and even enterococci (2, 4,
11). However, our hybridization studies demonstrated that
“S. suis” is not closely related to group D streptococci
(DNA relatedness values between strains of “S.suis” and
Enterococcus faecalis were less than 10%). This is consis-
tent with the numerical taxonomic studies of Bridge and
Sneath (1) and the opinion of other authors who do not
consider the reaction with group D antiserum a specific
characteristic for the identification of “S. suis” (3, 5).
As a result of this study it is confirmed that “ S . suis” is a
genetically homogeneous species. It has not yet been validly
described and is not on the Approved Lists of Bacterial
Names (14). On the basis of our results, the descriptions
given for group R and group S streptococci (2), and the
characterizations given by other authors for R, S, RS, and T
streptococci or “S.suis” (1,3,5,10, l l ) , we propose S. suis
as a valid species of the genus Streptococcus.
Description of Streptococcus suis (ex Elliott 1966) nom. rev.
Streptococcus suis (su’is. L. n. sus, a pig; L. gen. n. suis, of
a pig). Small ovoid cocci, less than 2 pm in diameter, occur
singly, in pairs, or rarely in short chains. Some tendency
toward rod formation. Nonmotile. Gram positive. Chemo-
organotrophs with fermentative metabolism. Facultatively
anaerobic. Catalase negative. Strains are groupable into
Lancefield groups R, S, RS, and T or are nongroupable.
They can also be subdivided into serovars 1to 8 according to
the method of Windsor and Elliott (15) and Perch et al. (11).
Acid from fermentation of D-glucose, sucrose, lactose,
VOL. 37, 1987 STREPTOCOCCUS SUZS sp. nov., NOM. REV. 161

TABLE 1. Organisms studied, serological and cell wall data, and G + C contents
Lance- Lysine Cell wall polysaccharides containing‘
Organism Strain” field Serovar peptido- mol%
group glycan typeb Rhamnose Glucose Galactose GlcNH2 GalNH2 G + C
“S. suis” NCDO 2644 ( = NCTC 10237) S Direct ND ND ND ND 40.7
“S. suis” Henrichsen S428 ( = NCTC S Direct ND ND ND ND ND
10237)
“S. suis” Henrichsen S735 ( = NCTC R Direct + + + + 40.6
10234)
“S. suis” Henrichsen 4961 Direct ND ND ND ND 39.9
“S. suis” Henrichsen 6407 Direct + (+I + (+I 39.1
“S. suis” Henrichsen 11538 (AMl-2 + + + + 40.8
“S. suis” Henrichsen 2524 Direct + + + + 41.0
“S. suis” Henrichsen 8074 Direct + + + + 40.3
“ S . suis” Henrichsen 14636 Direct + + + + 40.2
“ S . suis” Henrichsen 2651 RS (Awl-2 + + - + 42.0
‘S. suis” CCUG 7984 (= NCTC 10234) R Direct + + + + 38.4
“S. suis” CCUG 7985 (= NCTC 10237) S Direct ND ND ND ND ND
“S. suis” CCUG 7986 (= NCTC 10446) T Direct + + + + 40.0
“ S . suis” CCUG 11673 Direct + + + + 39.9
“S. viriduns” 111 Kiel45527 Direct + + +- (+I 40.0
S . pyogenes NCTC 8198T A (Aldl-3 + - + 38.0
S . oralis NCTC 11427T Direct (+I (+I + (+I 37.0
aNCDO, National Collection of Dairy Organisms, Shinfield, Reading, United Kingdom; Henrichsen, Streptococcus Department, Statens Serum Institut,
Copenhagen, Denmark; CCUG, Culture Collection University of Goteborg, Sweden; Kiel, Streptokokkenzentrale, Kiel, Federal Republic of Germany; NCTC,
National Collection of Type Cultures, Central Public Health Laboratory, London, United Kingdom.
Abbreviations of peptidoglycan types described by Schleifer and Kandler (13).
GlcNH2, Glucosamine; GalNH2, galactosamine; -, absent; + , present; (+), present in minor amounts; ND, not determined.

TABLE 2. DNA-DNA relatedness values obtained at optimal hybridization conditions (25°C below the melting temperature of DNA)
% Relatedness with labeled DNA from:
Source of
filter-bound DNA “S. suis” “S. suis” type 4 “S. suis” “S. suis” S. oralis
NCDO 2644 Henrichsen 6407 CCUG 7986 CCUG 11673 NCTC 11427T
“S. suis” NCDO 2644 100 93 19
“S. suis” type 2 Henrichsen S735 85 89 77 82
“S. suis” type 3 Henrichsen 4961 81 75 80
“S. suis” type 4 Henrichsen 6407 81 100 76 16
“S. suis” type 5 Henrichsen 11538 91 94 22
“S. suis” type 6 Henrichsen 2524 90 73 21
“ S . suis” type 7 Henrichsen 8074 89 98
“S. suis” type 8 Henrichsen 14636 93 100
“S. suis” Henrichsen 2651 100 89 20
“S. suis” CCUG 7984 97 77 100 80 19
“S. suis” CCUG 7985 100 75 83 86
“S. suis” CCUG 7986 81 88 100 92
“S. suis” CCUG 11673 100 84 100 20
“S. viriduns” 111 Kiel 45527 40 39 20
S. oralis NCTC 11427T 15 100
S . pyogenes NCTC 8198= 14 16 15 14 17

maltose, salicin, trehalose, and inulin. No fermentation of
L-arabinose, D-mannitol, D-sorbitol, glycerol, melezitose, or
D-ribose. Variable results in fermentation of raffinose and
melibiose and in production of hyaluronidase. Hydrolysis of
L-arginine, esculin, salicin, starch, and glycogen; no hydro-
lysis of hippurate; no production of acetoin. Acid phospha-
tase and alkaline phosphatase negative; L-ornithine
decarboxylase , N-acetylglucosaminidase, a-galactosidase ,
P-glucuronidase , and leucine arylamidase positive; p-
galactosidase variable. Resistant to optochin. No growth at
10 or 45”C, in 6.5% NaCl or 0.04% tellurite. Some strains are
resistant to 40% bile (5). Many strains produce P-hemolysis
on horse blood agar, and all are a-hemolytic on sheep blood
agar. Type of peptidoglycan is usually lysine direct, in a few
cases also lysine-(alanine)l-2. Cell wall polysaccharides con-
tain rhamnose, glucose, galactose, and glucosamine. G +C
content of DNA is 38 to 42 mol% (T,,J. Important pig
pathogen. The type strain is NCTC 10234 (= CCUG 7984 =
Henrichsen S735).
ACKNOWLEDGMENTS
We are grateful to J. Henrichsen, Statens Serum Institute,
Copenhagen, Denmark, for providing us with strains of serovars 1to
8.
This work was supported by a grant from the Deutsche
Forschungsgemeinschaft .
LITERATURE CITED
1. Bridge, P. D., and P. H. A. Sneath. 1983. Numerical taxonomy
of Streptococcus. J. Gen. Microbiol. 129565-597.
2. Deibel, R. H., and H. W. Seeiey, Jr. 1974. Genus I. Streptococ-
1.62 KILPPER-BALZ AND SCHLEIFER
cus Rosenbach 1884, 22, p. 490-509. In R. E. Buchanan and
N. E. Gibbons (ed.), Bergey's manual of determinative bacte-
riology, 8th ed. The Williams & Wilkins Co., Baltimore.
3. De Moor, C. E. 1963. Septicaemic infections in pigs, caused by
haemolytic streptococci of new Lancefield groups designated R,
S and T. Antonie van Leeuwenhoek J. Microbiol. Serol.
29:272-280.
4. Elliott, S. D. 1966. Streptococcal infection in young pigs. I. An
immunochemical study of the causative agent (PM streptococ-
cus). J. Hyg. 64205-212.
5. Hommez, J., L. A. Devriese, J. Henrichsen, and F. Castryck.
1986. Identification and characterization of Streptococcus suis.
Vet. Microbiol. 11:349-355.
6. Kilpper, R., U. Buhl, and K. H. Schleifer. 1980. Nucleic acid
homology studies between Peptococcus saccharolyticus and
various anaerobic and facultatively anaerobic Gram-positive
cocci. FEMS Microbiol. Lett. 8:205-210.
7. Kilpper-Balz, R., and K. H. Schleifer. 1981. DNA-rRNA hybrid-
ization studies among staphylococci and some other Gram-
positive bacteria. FEMS Microbiol. Lett. 10:357-362.
8. Kilpper-Balz, R., P. Wenzig, and K. H. Schleifer. 1985. Molec-
ular relationships and classification of some viridans strepto-
cocci as Streptococcus oralis and emended description of Strep-
tococcus oralis (Bridge and Sneath 1982). Int. J. Syst. Bacteriol.
INT. J. SYST.BACTERIOL.
35482488.
9. Kilpper-Balz, R., B. L. Williams, R. Lutticken, and K. H.
Schleifer. 1984. Relatedness of "Streptococcus milieri'' with
Streptococcus anginosus and Streptococcus constellatus. Syst.
Appl. Microbiol. 5494-500.
10. Perch, B., E. Kjems, P. Slot, and K. B. Pedersen. 1981. Bio-
chemical and serological properties of R, S, and RS strepto-
cocci. Acta Pathol. Microbiol. Scand. Sect. B 89:167-171.
11. Perch, B., K. B. Pedersen, and J. Henrichsen. 1983. Serology of
capsulated streptococci pathogenic for pigs: six new serotypes
of Streptococcus suis. J. Clin. Microbiol. 17:993-996.
12. Schleifer, K. H., and 0. Kandler. 1967. Zur chemischen Zusam-
mensetzung der Zellwand der Streptokokken. I. Die Amino-
sauresequenz des Mureins von Streptococcus thermophilus and
Streptococcus faecalis. Arch. Mikrobiol. 57:335-364.
13. Schleifer, K. H., and 0. Kandler. 1972. Peptidoglycan types of
bacterial cell walls and their taxonomic implications. Bacteriol.
Rev. 36:407477.
14. Skerman, V. B. D., V. McGowan, and P. H. A. Sneath. 1980.
Approved lists of bacterial names. Int. J. Syst. Bacteriol.
30:225420.
15. Windsor, R. S., and S. D. Elliott. 1975. Streptococcal infection
in young pigs. IV. An outbreak of streptococcal meningitis in
weaned pigs. J. Hyg. 7569-78.