Streptococcus suis Infection in Swine.

A Sixteen Month Study
R. Higgins, M. Gottschalk, K.R. Mittal and M. Beaudoin

fois. Quarante et un pourcent des isolats typables et 33% des nonA total of 349 isolates of Strepto- typables furent isoles en culture pure. coccus suis retrieved from different Les autres isolats etaient retrouves tissues from diseased pigs were avec Pasteurella multocida, Escheriexamined in this study. Only 48% of chia coli, Actinobacillus pleuropneuthem could be categorized as one of moniae, Actinomyces pyogenes et serotypes 1 to 8 and 1/2. Among d'autres streptocoques. Les isolats typable isolates, serotype 2 was the typables provenaient le plus souvent most prevalent (23%), followed by de porcs ages de cinq a dix semaines, serotype 3 (10%). The majority of all alors que les non-typables etaient isolates originated from lungs, men- isoles d'animaux plus ages que 24 inges/brain, and multiple tissues. semaines. Aucune variation mensuelle Forty-one percent of typable isolates et/ou saisonniere de l'isolement de S. and 33% of untypable isolates were suis n'a pu etre detectee. retrieved in pure culture. Other isolates were found in conjunction INTRODUCTION with Pasteurella multocida, Escherichia coli, A ctinobacillus pleuropneuStreptococcus suis is an important moniae, Actinomyces pyogenes, and other streptococci. Typable S. suis pathogen of swine in most countries isolates were more frequently isolated where the pig industry is developed. It from pigs between five and ten weeks has been associated mainly with of age, while untypable isolates were bronchopneumonia, meningitis and mostly found in animals aged more septicemia, and less frequently with than 24 weeks. No obvious monthly endocarditis, arthritis or other condiand/or seasonal variation of the tions (1-6). This microorganism shares prevalence of isolation of S. suis could cell wall antigens with other members of group D of Lancefield (7), but is be detected. genetically not related to them (8). Originally, three serotypes, 1, 2 and 1/2 were recognized, based on the RESUME antigenicity of the capsular material Sur un total de 349 isolats de (7,9). These serotypes corresponded to Streptococcus suis provenant de porcs serogroups S, R and RS respectively, soumis pour une necropsie, seulement as described earlier by de Moor (10). 48% ont pu etre associes a l'un des In 1983, Perch et al (1 1) presented six serotypes 1 a 8 et 1/2. Parmi les isolats new serotypes of S. suis (3 to 8), all ayant ete serotypes, le serotype 2 s'est implicated in a variety of lesions. In Canada, there have been several avere le plus prevalent (23%), suivi du serotype 3 (10%). La plus grande reports on the importance of S. suis partie de tous ces isolats de S. suis serotype 2 and the pathological provenaient des poumons, des menin- changes associated with this microges/cerveau et de plusieurs tissus 'a la organism (4,12-15). The only study

ABSTRACT

which involved all nine serotypes was from Quebec and it consisted of a retrospective study of the importance of S. suis over a nine year period (6). The purpose of the present study was to determine the actual prevalence of the nine recognized serotypes among biochemically identified S. suis isolates, collected from diseased animals during a 16-month period. Information about the site of isolation, age of animals, and seasonal incidence is presented and discussed.
MATERIALS AND METHODS
BACTERIAL STRAINS

types

Reference strains of S. suis sero1 to 8 and 1/2 were kindly supplied by Dr. Jorgen Henrichsen of the Statens Seruminstitut in Denmark. A total of 349 isolates of S. suis were collected between August 1987 and November 1988 from diseased pigs submitted for necropsy at the College of Veterinary Medicine of the University of Montreal, and at the provincial veterinary laboratories at SaintHyacinthe and Sainte-Foy, Quebec. Tissue specimens were cultured on tryptic soy agar (Difco Laboratories, Detroit, Michigan) with 5% bovine blood. Plates were incubated at 370C with 5% CO2. Only streptococci isolated in pure culture or present in a significant number, were submitted for further studies.
BIOCHEMICAL IDENTIFICATION

Alpha-hemolytic streptococci retrieved from different animals were originally subjected to four biochemi-

Groupe de Recherche sur les Maladies Infectieuses Porcines, Faculte de Medecine veterinaire, Universite de Montreal, C.P. 5000, Saint-Hyacinthe (Quebec) J2S 7C6. Submitted March 22, 1989.

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Can J Vet Res 1990; 54: 170-173

suis was Isolation of typable S.4. Of the 349 isolates biochemically Other isolates were found in conjuncidentified as S.T. NaCl and acetoin negative. mannitol and production of acetoin. according to the tissues or organs from which they originated Origin aUntypable No. an isolate biochemically compatible with S.5% of of 182 (52%) isolates were untypable. The pleuropneumoniae. trehalose.Compared to other serotypes. suiSa S. Distribution of typable and untypable isolates of Streptococcus suis. suis were serotyped by the coagglutination technique (16). A total aged more than 24 weeks.were untypable except one of serotype strated that all isolates which did not 5 and one of serotype 6. All untypable isolates were tested by the following 11 tests: growth in 6. RESULTS In fetuses. The (possible septicemia) (Table II).b 6 5 4 3 2 67 9 6 2 8 26 36 Lungs 1 1 51 1 1 5 10 Meninges/ brain 31 3 1 1 1 19 Multiple tissuesc 1 18 1 2 5 Heart 3 2 2 Joints 3 1 1 2 Genital tract 24 2 3 2 1 3 3 1 5 Others aNumber of isolates (some isolates were retrieved in two different sites) bU. A positive reaction was characterized by a distinct clumping.T. only 6. = Untypable clsolates found in three or more different tissues. glycerol. suis were retrieved in pure culture S.5) 8 (2) 79 (23) 35 (10) 11 (3) 3 (1) 7 (2) 10 (3) 12 (3. Twenty-three percent of all isolates were of serotype Escherichia coli. One drop of the coagglutination reagent was mixed on a glass slide with an equal volume of bacterial suspension. typable isolates were retrieved from The majority of S. SEROTYPING Streptococci biochemically identified as S. suis was defined as follows: absence of growth in the presence of NaCl. A ctinobacillus 2 and 10% were serotype 3.a No.TABLE I. The coagglutination test is routinely used for serotyping S. Controls consisted of nonlabelled staphylococcal cell suspension. suis. Fetuses were included in this category 1 1 1/2 5 2 2 Total 160 72 58 27 7 7 44 cal tests in diagnostic laboratories: growth in 6. usually occurring within a few seconds. as well as staphylococcal cell suspension coated with normal rabbit serum.5%) and three (1%) isolates were mostly found in animals isolates respectively (Table I). were sent to our research laboratory for serotyping. of isolates of S. The prevalence of serotypes 1 and 5 was the and ten weeks of age. Tests were performed as previously described (6). against serotypes 1 to 8. sucrose. and trehalose and salicin positive. and seven of the nine tests with a reaction as expected. were confirmed as S. followed by serotype 1 isolates were found in meninges/ brain and multiple tissues animals less than one week old. Distribution of the 349 Streptococcus suis isolates. Prevalence of different bacterial microorganisms isolated in conjunction with Streptococcus suis in tissues from diseased pigs No.17). production of acid from trehalose and salicin. suis by all the other biochemical from the different tissues (Table III).14. serotype 2 as well as untypable isolates dered positive.T.5 or less. acetoin negative. Reactions of 2+ or more were consi. this age belonged to serotype 8. suis in North America (1. All isolates. sorbitol. suis with: No other bacterium (pure culture) Typable 69 (41%) 49 27 Untypable 60 (33%) 28 Pasteurella multocida 48 Escherichia coli 11 14 Actinobacillus pleuropneumoniae 11 10 Actinomyces pyogenes 10 1 Other streptococci 30 15 Other bacteria aSome isolates were retrieved in conjunction with two or three different bacterial microorganisms 171 . grow in the presence of NaCl and were Forty-one percent of typable isoacetoin negative. all isolates of S. The reaction was recorded within 3 min and was scored on a 0 to 4+ basis depending on the rapidity and intensity of the reaction. according to serotyping Serotype 1 1/2 2 3 4 5 6 7 8 U. of isolates (%) 2 (0. and production of acetoin. arginine dihydrolase.5% NaCl. tests. lactose.5) 182 (52) TABLE II. suis isolates this age group (Table IV). Actinomycespyoremainder of the typable isolates were genes and other streptococci. distributed among the other serotypes more frequent in animals between five with percentages of 3. of isolates by serotypea 8 7 U.5% NaCl. inulin. TABLE III. suis were retrieved from multiple tissues and Biochemical identification demon. only 48% could be tion with the common bacterial serotyped with one of the antisera pathogens: Pasteurella multocida. as well as salicin and lates and 33% of untypable isolates of trehalose positive. were more frequently isolated from multiple tissues and meninges/brain. Antisera were prepared in rabbits against reference strains of serotypes 1 to 8. The two originated from lungs. production of acid from salicin. Because of possible variations in test reactions. other typable isolate from piglets at Serotype 2 was present in all sites.15. while untypable lowest with two (0.

A. suis was monitored over a oneyear period. multocida. All nine recognized serotypes were found among our typable isolates.5 42 28. The absence of serotype 1 in four to eight week old piglets has also been confirmed by us (unpublished data). DISCUSSION This study demonstrated that the absence of growth in the presence of 6. This study was not designed as a prevalence study. TABLE V. Interestingly. of isolates 28 20 16 20 15 23 (%) (10. More significantly. The prevalence of typable S. This appears to be an indication of the greater pathogenicity or virulence of this serotype over others. This percentage was 60% for untypable isolates. suis serotype 2 cases in the colder and hence more environmentally confining months of the year.5) (12) (7. The number of untypable isolates had generally a prevalence proportional to that of typable isolates in other tissues. were untypable (6).5) (8. More than half of the 349 isolates (52%) of S. Also. suis isolates. the Conseil des Recherches en Peche et Agro-alimentaire du Quebec (2021). This work was supported in part by the Fonds pour la Formation de Chercheurs et l'Aide a la Recherche (89 EQ 4101). The finding that 41% of typable isolates and 33% of untypable isolates of S. and the Ministere de l'Enseignement superieur et de la Science.5 5. This could indicate that some serotypes are related to certain age groups as is serotype 1.19. suis infections. This agrees with a report from Iowa (25). The high prevalence of S. and they have undertaken studies on the pathogenicity of this serotype (21). ERICKSON ED. Also. suis Typable Untypable 2 1 14 15. are sufficient for a presumptive identification of S. The serotyping of all the so-called untypable isolates is a prerequisite for further studies on epidemiology.23). the failure to produce acetoin. from August 1987 to July 1988 (Table V).5) Month No. and it reinforces the importance of the establishment of an official serotyping system. The number of untypable isolates from multiple tissues and meninges was important and this could include some new serotypes whose virulence could be comparable to that of serotype 2. The presence of microorganisms in conjunction with the isolation of S. 75% of typable isolates were from pigs between one and 18 weeks of age. Sylvie Lacasse and Marie-Andree Lussier. pleuropneumoniae. and the production of acid from salicin and trehalose.13. so statistical analysis could not be performed and it was not possible to determine the presence or absence of monthly and/ or seasonally significant variations.1 1). suis was meninges.5) (9) (10.5) 172 . Contrary to a report from the United States (24) in which the most common source of S.5 The monthly incidence of isolation of S. But other reports from England (27) and the United States (24) indicated that there was no seasonal incidence of the disease. this study did not detect any particular monthly and/ or seasonal variations in the number of S. This is in accordance with previous results obtained in Quebec (6).22. suis isolates belonged to the above serotypes (3).5 5 3 6. suis by pathologists. over a nine year period.5 46 1 5 9. and to a systematic search in diagnostic laboratories. suis could not be serotyped with antisera against the recognized serotypes 1 to 8.TABLE IV. 185: 666-668. pyogenes and others) has also been well documented (3. the total number of isolates from lungs outnumbered those from other sites.13). This agrees with the previous reports (5. This is more than in an earlier report from our laboratory in which 24% of isolates from diseased pigs.6. suis isolations. suis (A. Monthly incidence of isolation of Streptococcus suis over a one-year period (August 1987 to July 1988) Month August 1987 September 1987 October 1987 November 1987 December 1987 January 1988 No. suis. Finally. 27% and 28% respectively. REFERENCES 1. Other reports from Canada (18). suis were recovered in pure culture from the different specimens cultured agrees with previous results (6). One Canadian report (13) mentioned that there was an apparent concentration of S. suis serotype 2 in diseased pigs is also consistent with other reports (3. The highest rate of isolation was in June 1988 (12%) and the lowest rate was in February 1988 (5%). ACKNOWLEDGMENTS The authors would like to acknowledge the invaluable assistance of Monique Gaumond.20).5 20 16. Distribution of Streptococcus suis isolates according to the age of animals Age (weeks) <1 1 to 4 5 to 10 llto 18 19to24 > 24 Fetuses Not determined Percentage of isolates of S. Scandinavian workers showed a higher prevalence of serotype 7 over all others including serotype 2 (5. P. and 1 / 2. DOSTER AR.5) February 1988 March 1988 April 1988 May 1988 June 1988 July 1988 28 24 28 33 20 (5) (10.5) (5. suis isolates was greater in pigs aged from five to ten weeks. of isolates 14 (%) (7.6. Isolation of Streptococcus suis from swine in Nebraska. Belgium (19) and the United States (20) reported percentages of untypable isolates of 64%. J Am Vet Med Assoc 1984. POKORNY TS.5% NaCl. The increase in the number of untypable isolates could be attributed to a greater awareness of S.5) (6) (7. diagnosis and control of S. authors from France have reported that 197 of 200 S. Power (26) noted that the number of meningitis cases increased with sudden changes in weather. The percentage of untypable isolates originating from pigs aged more than 24 weeks was almost eight times higher than that of typable S.

HENDERSEN LM. 17. PERCH B. LAMONT MH. 22. VAN LEENGOED LAMG. HENRICHSEN J. 9. SANFORD SE. Serology of capsulated streptocci pathogenic for pigs: Six new serotypes of Streptococcus suis. Proc Am Assoc Vet Lab Diagnost 1983. 11. 3. 51: 481485. 20. Can J Vet Res 1987. Rev. Central nervous system lesions. An outbreak of streptococcal meningitis in weaned pigs. An immunochemical study of the causative agent (PM Streptococcus). Can J Vet Res 1986. 7: 315-321. 24. ST JOHN VS. KOTT B. Antonie van Leeuwenhoek 1963. HOLTER JA. 23: 95-97. KIERSTEAD M. MITCHELL WR. BRETON J. Streptococcus suis type 2 associated diseases in swine: Observations of a one-year study. CORNELL WD. Rec MMd Vet 1985. Isolation and serotype prevalence of Streptococcus suis in seven midwestern states. 102: 215-216. Infection porcine a Streptococcus suis dans le departement des C6tes-du-Nord. 40: 1640-1641. nov. pathologic and bacteriologic data from a two year study. Can J Vet Res 1987. DEVRIESE LA. 161: 353-359. Vet Q 1985. 18: 1351-1354. 16. 50: 338-341. 15. J Clin Microbiol 1983. 12. Streptococcus suis infections in pigs in the Netherlands (part I). TOUIL F. CASTRYCK F. ROSENDAL S. Can Vet J 1982. 29: 272-280. 64: 205-212. 173 . MADDUX RL. 26.2. Proc Am Assoc Swine Pract 1989: 365-367. Streptococcus suis type 2 infections in pigs. Vet Rec 1980. 75: 69-78. Streptococcal meningitis in pigs: Results of a five-year survey. J Hyg 1966. Acta Vet Scand 1988. Am J Vet Res 1979. Cardiac lesions. WINDSOR RS. SANFORD SE. 4. WERHEIJEN ERM. Iowa State University Veterinary Diagnostic Laboratory. MITTAL KR. BRETON J. Gross and histopathological findings in unusual lesions caused by Streptococcus suis in pigs II. NADEAU M. Methodes d'identification et repartition des differents serotypes rencontres. Vet Rec 1978. KOEHNE G. S and T. 181: 673-676. 5. DE MOOR CE. Int J Syst Bacteriol 1987. HENRICHSEN J. Proc Am Assoc Vet Lab Diagnost 1985. 8. 107: 467-469. 10. J Hyg Camb 1975. BOETNER AG. WINDSOR RS. 26: 393-402. MITCHELL WR. 7. ELLIOTT SD. VECHT U. J Am Vet Med Assoc 1982. 19. GINDER M. 51: 486-489. Nom. Streptococcus suis infections in Danish pigs and experimental infection with Streptococcus suis serotype 7. 17: 171-177. ROSENDAL S. SCHLEIFER KH. ELLIOT SD. Meningitis in pigs caused by Streptococcus suis type II. 27. HENRICHSEN J. Isolation of Streptococcus suis from diseased pigs in Canada. Streptococcus suis type 2 infection in swine in Ontario: A review of clinical and pathological presentations. HOFFMAN LJ. Streptococcus suis in slaughter pigs and abattoir workers. Cases of swine pneumonia and meningitis associated with Streptococcus suis. 26: 121-130. LE MENEC M. LARSON DJ. 11: 349-355. J Clin Microbiol 1983. 18. The significance of Streptococcus suis in swine disease: Clinical. HIGGINS R. Proc Am Assoc Vet Lab Diagnost 1984. Vet Microbiol 1986. Can J Vet Res 1986. Identification and serotyping of Haemophilus pleuropneumoniae by coagglutination test. Streptococcus suis isolated from pigs in Finland. HIGGINS R. 13. 37: 160-162. 14. 6. HOMMEZ J. Streptococcal infections in young pigs. BILLEHANSEN V. I. Lancefield group R streptococci associated with pneumonia in swine. KOBISCH M. KILPPER-BALZ R. 23. 101: 378-379. 29: 9-13. Gross and histopathological findings in unusual lesions caused by Streptococcus suis in pigs I. HENRICHSEN J. SANFORD SE. IV. POWER JB. Septicaemic infections in pigs caused by haemolytic streptococci of new Lancefield groups designated R. 50: 537-539. Streptococcus suis sp. 25. 95: 233-239. SANDERS P. Identification and characterization of Streptococcus suis. TILKER AME. 17: 993-996. Vet Rec 1977. WILKINS ML. WINDSOR RS. 21. Vet Microbiol 1988. EDWARDS PT. HILT L. PEDERSEN KB.. WILCOCK B. KURL DN. SIHVONEN L. 28: 201-2 10. LARIVIERE S. Streptococcal infection in young pigs. Acta Pathol Microbiol Immunol Scand [B] 1987. Purulent meningitis in Iowa swine. Isolation of Streptococcus suis using a selective medium.