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Streptococcus Canis Sp. Nov.: a Species of Group G Streptococci from Animals

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Streptococcus Canis Sp. Nov.: a Species of Group G Streptococci from Animals INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, July 1986, p. 422425 Vol. 36, No. 3 0020-7713/86/03O422-04$02.OO/O Copyright 0 1986, International Union of Microbiological Societies Streptococcus canis sp. nov.: a Species of Group G Streptococci from Animals LUC A. DEVRIESE ,l* JOZEF HOMMEZ,’ RENATE KILPPER-BALZ,3 AND KARL-HEINZ SCHLEIFER3 Faculty of Veterinary Medicine, University of Gent, B-9000 Gent, Belgium’; Regional Veterinary Investigation Centre, Torhout, Belgium2; and Lehrstuhl fur Mikrobiologie, Technische Universitat, Munich, Federal Republic of Germany3 f3-Hemolytic group G streptococci from cows with mastitis and from dogs with different pathological conditions were characterized and named Streptococcus canis sp. nov. These animal group G strains were differentiated from Streptococcus dysgalactiae (group G strains of human origin, as well as group L strains and certain animal and human group C strains) by simple physiological and biochemical tests. Deoxyribonucleic acid-deoxyribonucleic acid hybridization showed that the S. canis strains differed from S. dysgalactiae, Streptococcus pyogenes, Streptococcus equi, and Streptococcus salivarius. These organisms contain a rare cell wall peptidoglycan type, namely, Lys-Thr-Gly. The specific epithet of S. canis, which has been in use for many years in veterinary textbooks to designate group G strains isolated from dogs, was chosen for this proposed new species because it comprises animal but not human group G streptococci. The type strain is strain STR-T1 (= DSM 20715). Streptococci belonging to Lancefield group G (15) form a Clark et al. (6), led us to examine the taxonomic status of heterogeneous collection of organisms. At least the follow- animal group G streptococci. ing three distinct groups can be found: (i) the large-colony form of P-hemolytic group G streptococci isolated from MATERIALS AND METHODS humans; (ii) large-colony P-hemolytic strains from animals which were first shown to differ from human group G strains Strains. A total of 28 group G strains from cases of bovine in their fibrinolytic activity (3) and later also in several other mastitis on 28 different farms were isolated in 1982, 1983, characteristics (6); and (iii) the “minute P-hemolytic col- and 1984; these strains were designated strains STR-T1, ony” group G strains from humans. STR-T2, STR-T3, STR-T4, STR-T5, STR-T7, STR-T32, Animal group G strains have been associated mainly with STR-T35, STR-T37, STR-T38, STR-T40, STR-T41, STR- genital, skin, and wound infections (2, 10) in dogs and with T42, STR-T43, STR-T44, STR-T45, STR-T47, STR-T50, bovine mastitis (6, 16). The naming of these strains has STR-T51, STR-T52, STR-T53, STR-T54, STR-T55, STR- always been confusing and contradictory. Stafseth et al. (20) T56, STR-T57, STR-T70, STR-T71, and STR-T72. The fol- first used the name “Streptococcus canis” to designate dog lowing three group G strains were isolated from dogs: strain strains belonging to group C. However, these strains had a STR-290 from wound exudate and strains STR-312 and characteristic carbohydrate fermentation pattern which was STR-313 from neonatal septicemia cases. A human group G found later only in animal group G strains (2). Widely used strain (strain G/D 166B) was obtained from D. Groothuis, veterinary textbooks (4,5) refer to group G strains from dogs Public Health Institute, Bilthoven, The Netherlands. Ten as “Streptococcus canis,” but this name has never been group L strains and group G strains from cases of bovine accepted officially. mastitis were used in comparative antibiotic susceptibility Recent nucleic acid hybridization and cell wall composi- tests. S. dysgalactiae DSM 20662T (T = type strain), Strep- tion studies (13) have shown that human large-colony group tococcus equi DSM 20561T, Streptococcus salivarius ATCC G strains are closely related to group L strains, the P- 13419 and ATCC 20560T, and Streptococcus pyogenes hemolytic group C organism “Streptococcus equisimilis,” NCTC 819gT, which were obtained from the Deutsche and a-hemolytic group C strains isolated from cases of Sammlung von Mikroorganismen (Gottingen, Federal Re- bovine mastitis and polyarthritis in lambs. Similar observa- public of Germany), the National Collection of Type Cul- tions led to an emended description of Streptococcus tures (Colindale, United Kingdom), and the American Type dysgalactiae (9) which fits these animal group C P-hemolytic Culture Collection (Rockville, Md.) were used in deoxyribo- “S. equisimilis,” group C a-hemolytic S. dysgalactiae, and nucleic acid (DNA)-DNA hybridization tests. group L strains, as well as human group C “S. equisimilis,” Physiological tests. Growth characteristics were investi- group L, and group G strains. The older description of S. gated in brain heart infusion medium (Oxoid Ltd., dysgalactiae given in Bergey ’s Manual of Determinative Basingstoke, United Kingdom), in Todd-Hewitt broth (Ox- Bacteriology, 8th ed. (7), applied only to the animal a- oid), and on 5% sheep blood agar containing tryptose blood hemolytic group C strains. agar base (Oxoid). Resistance to 6.5% NaCl was tested in However, it has been found that several characteristics of brain heart infusion medium, and resistance to 40% bile was animal group G strains in our collection did not correspond tested in bile esculin agar (Difco Laboratories, Detroit, to the characteristics given in the new description of S. Mich.) which was incubated for 48 h. Formation of acid, dysgalactiae. These observations, as well as the description reduction, and coagulation of litmus milk was studied in of the biotypes of animal and human group G strains by litmus milk medium (Difco). Decarboxylation of L-tyrosine was tested as described by Sharpe (19). Hyaluronidase activity was determined on blood agar that was swab inoc- ulated with a Pasteurella multocida capsule type A strain * Corresponding author. and fibrinolysin on nutrient agar (Oxoid) containing 28 mg of 422 VOL. 36, 1986 STREPTOCOCCUS CANIS SP. NOV. 423 TABLE 1. Characteristics of 31 S. canis strains" human fibrinogen per liter or 6 mg of bovine fibrinogen per No. of strains liter. Pyrrolidonylarylamidase and P-glucuronidase activities Characteristic positivelno. of were tested by using media and reagents formulated by API strains tested System (La Balme les Grottes, France) and Rosco Catalase activity ................................. 013 1 (Taastrup, Denmark), and L-alanine aminopeptidase activity Growth in 6.5% NaCl ............................ 013 1 was tested with Bactident Aminopeptidase (E. Merck AG, Growth in 40% bile .............................. 013 1 Darmstadt , Federal Republic of Germany). Esculin degrada- CAMP reaction .................................. 013 1 tion was tested by using a API-20 Strep kit (API) and on agar Hippurate reduction. ............................. 013 1 slants containing (per liter) 0.1 g of esculin, 0.1 g of ammo- Esculin degradation in agar medium ............. 31/31 nium ferric citrate, 10 g of tryptone, 5 g of peptone, 5 g of Esculin degradation in API ...................... 31 (27 weak)/31 yeast extract, and 15 g of agar (pH 7.3). Starch hydrolysis Fibrinol y sin ...................................... 013 1 was evaluated by inoculating Mueller-Hinton agar (Oxoid) Tyrosine decarboxylation ........................ 013 1 and flooding the plates with iodine after 24 h. Hippurate Hyaluronidase activity ........................... 1 (weak)/31b Starch hydrolysis ................................ 013 1 hydrolysis, the Voges-Proskauer reaction, a- and P-galac- L-Alanine aminopeptidase activity ............... 15/15 tosidases, alkaline phosphatase, leucine arylamidase, argi- Voges-Proskauer test ............................ 013 1 nine dehydrolase, and acid production from carbohydrates Alkaline phosphatase activity .................... 3 113 1 were tested by using API-20 Strep and API-50 CH systems. Leucine arylamidase activity. .................... 3 113 1 Antibiotic susceptibility testing. Overnight brain heart infu- Arginine dehydrolase activity .................... 3113 1 sion cultures were diluted 20-fold in buffered saline and Pyrrolidonylarylamidase activity ................. 013 1 inoculated onto Mueller-Hinton agar (Oxoid) and onto Iso- a-Galactosidase activity.. ........................ 2313 1' Sensitest agar (Oxoid) supplemented with 5% bovine blood P-galactosidase activity .......................... 28/31' or laked horse blood and doubling dilutions of penicillin G, P-glucuronidase activity. ......................... 4/31' lincomycin hydrochloride, or trimethoprim. The results Acid produced from: N-Acet ylglucosamine .......................... 919 were read after 24 h at 37°C. These antibiotics were chosen Adonitol ....................................... 019 for quantitative testing because agar diffusion tests carried Am ygdaline .................................... 019 out on Iso-Sensitest agar enriched with 7% lysed horse blood L- Arabinose .................................... 013 1 by using Rosco susceptibility test tablets showed that the D- Arabinose ................................... 019 inhibition zone diameters of these antibiotics were larger D- Arabitol ..................................... 019 with the animal group G strains than with group L S. Arbutin ........................................ 919 dysgalactiae strains. Cellobiose ..................................... 819' Serological tests. For serological tests we used the capil- Dulcitol ........................................ 019
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