Tests useful for the identification of Enterococcus strains were applied to a collection of isolates from animal intestines and to reference strains, all of which were capable of growth on 40% bile and in 6.5% NaCl. Most strains could be identified as known species, and their characteristics corresponded, with a few exceptions of minor importance, with those described for Enterococcus hirae, Enterococcus durans, Enterococcus mundtii, Enterococcus gallinarum, Enterococcus avium, and Enterococcus casseliflavus. However, some diagnostically important carbohydrate reactions of Enterococcus faecalis and Enterococcus faecium strains differed from those given in the species descriptions and in recent reports. Production of acid from D-raffinose and D-xylose by E . faecium varied with the host species from which the strains were isolated. E . durans and E. gallinarum were isolated only from poultry, whereas E. avium was found only in mammals.Enterococci are an important group of intestinal bacteria whose taxonomy has undergone important changes in the last few years. The enterococci (sensu Sherman [9]), Streptococcus bovis, and Streptococcus equinus have been classified traditionally as serological group D streptococci. However, nucleic acid studies have shown that Streptococcus faecalis and Streptococcus faecium are only distantly related to S. bovis and S. equinus (5,7). These studies resulted in the proposal to transfer S. faecalis and S. faecium to a new genus, Enterococcus, as E. faecalis and E. faecium (8).Other group D streptococci which belong to the enterococcal MATERIALS AND METHODS Strains.A total of 264 strains (Table 1) were selected solely on the basis of their ability to grow in the presence of 40% bile (bile-esculin agar, Difco Laboratories, Detroit, Mich.) and in 6.5% NaCl in brain heart infusion agar (Oxoid Ltd., London, England). The selection was made from a larger collection of catalase-negative, gram-positive cocci and short rods isolated from the intestines of farm animals on mitis salivarius agar (GIBCO, Paisley, Scotland) or brain heart infusion agar (Oxoid) with 10 pg of oxolinic acid

Carboxyphilic strains which had characteristics commonly attributed to the group D streptococci and especially to Streptococcus bovis were isolated from the ceca of chickens. However, these strains differed from S. bovis in being positive for ribose, melezitose, P-glucuronidase, and alkaline phosphatase, and negative for group D reactions and growth enhancement by C02. These differences were confirmed by deoxyribonucleic acid-deoxyribonucleic acid hybridization tests. The strains form a new species, for which the name Streptococcus c'ecorum is proposed. Strain A60 (=NCDO 2674) is the type strain of this species.During an investigation of the susceptibilities of intestinal bacteria isolated from chickens to antimicrobial agents used in farm animals (G. N . Dutta, D. Vet. Sci. thesis, University of Gent, Gent, Belgium, 1983), a group of carboxyphilic streptococci was found which could not be identified as any known streptococcal species. The strains in this group were able to grow at 45°C and in the presence of 40% bile, two characteristics which are commonly attributed only to the enterococci or group D streptococci. After the tests advocated by Facklam (6) and Sharpe (21) for the separation of fecal streptococci from other streptococci (growth at pH 9.6, in 6.5% NaC1, and at 10 and 45°C) were performed, the strains were tentatively identified as Streptococcus bovis. However, three reactions were in discordance with this identification; the strains failed to react with the Streptococcus group D antisera used, they were able to grow to a limited extent at pH 9.6, and their growth was definitely better in an atmosphere containing 5% C02 than in air.A subsequent biochemical and genetic study showed that these strains should be designated a new species, which is described here. MATERIALS AND METHODSStrains. Strains A2a, A16, A29, A37 through A40, and A60T (T = type strain) were isolated from the ceca of eight chickens from eight farms in Belgium by using Islam medium (12) supplemented with 30 Fg of neomycin per ml and 15 pg of nalidixic acid per ml. The plates were incubated for 48 h in 5% COz in air. S . bovis CCM 5616 was obtained from the Czechoslovak Collection of Microorganisms, Brno. Other reference strains were obtained from the National Collection of Dairy Organisms (NCDO), Shinfield, Reading, England. _____Media and culture incubation. Media were obtained from Oxoid Ltd., Basingstoke, England, unless otherwise stated. Cultures were normally incubated at 37°C.Growth characteristics. Growth on sheep bloodColumbia agar plates was compared after incubation aerobically and in a 5% C 0 2 atmosphere for 16 and 48 h. All of the other tests except the API tests were performed in a 5 to 10% C 0 2 atmosphere.Biochemical reactions. Dextran and levan formation was tested on heart infusion agar supplemented with 5% (wthol) sucrose. Acid production was tested in phenol red broth base (GIBCO, Paisley, Scotland) containing 1% (wthol) carbohydrate. Glycerol fermentation was tested anaerobically on the solidified medium descri...

Strains isolated from pigeons belonging to the coagulase-positive species Staphylococcus intermedius, coagulase-negative Staph. hyicus subsp. chromogenes strains from cattle and pigs, and Staph. aureus strains from poultry, gave weakly positive reactions in DNase plate culture tests and heat-resistant DNase tests. Staph. aureus and Staph. intermedius strains from other sources and coagulase-negative and coagulase-positive Staph. hyicus subsp. hyicus strains reacted strongly in these tests. A standardized plate culture test procedure is proposed and the use of DNase tests in the identification of staphylococci isolated from animals is discussed. 002 1-8847/79/020385 + 09%01.00/0 13851 0 1979 The Society for Applied Bacteriology .i: Standard deviations.? z c m c t l rn E m c1 R z 0

Different nutrient bases, fibrinogen or heated plasma preparations and plasminogen sources were compared in tests for staphylokinase production by Staphylococcus aureus and other staphylococcal species. Media containing a nutrient base, bovine fibrin and dog plasma or serum proved to be more sensitive than the other substrates or combinations of substrates tested. Comparative testing on plates wih and without dog plasminogen or with plasmin inhibitors, was found to be necessary for the differentiation of staphylokinase from protease effects. Staphylokinases were produced by S. aureus, S. hyicus, S. simulans, S. sciuri and S. xylosus strains.

Reproducible and comparable results were obtained in different media containing different nutrients and indicators when representative Staphylococcus strains were tested for acid production from carbohydrates. Only in tests with melezitose, ribose and turanose many discrepant, weak or delayed positive reactions occurred. In tests with other carbohydrates similarly many weak results were visible after 1 day of incubation, but acid reactions were clear and easy to read after 2 days in phenol red broth and Andrade peptone water media, commonly used in the routine identification of other bacterial genera. Important alkalinization effects were observed after prolonged incubation, but not after short periods up to 24 h in peptone-rich test media. Growth was poor and numbers of acid reactions from carbohydrate breakdown detected did not increase when peptone-free media were used.