INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY,OCt. 1987, p. 310-316 Vol. 37, No. 4 0020-7713/87/04310-07$02.00/0 Copyright 0 1987, International Union of Microbiological Societies

Classification of divergens, Lactobacillus piscicola, and Some Catalase-Negative, Asporogenous, Rod-Shaped from Poultry in a New Genus, Carnobacterium

M. D. COLLINS,l J. A. E. FARROW,l B. A. PHILLIPS,l S. FERUSU,2 AND D. JONES2* Department of Food Microbiology, AFRC Institute of Food Research, Shinfield, Reading RG2 and Department of Microbiology, School of Medicine and School of Biological Sciences, University of Leicester, Leicester LEI 7RH,2 United Kingdom

Biochemical and chemical studies were performed on some atypical lactobacilli from chicken meat in an attempt to clarify their . The present study showed that the majority of the poultry strains could be allocated to the Brochothrix thermosphacta, Lactobacillus divergens, and Lactobacillus piscicola. The majority of the remaining poultry strains fell into two distinct groups which were worthy of separate species status. Lactobacillus carnis was found to be a member of the species L. piscicola. On the basis of biochemical, physiological, and chemical criteria, we suggest that L. divergens, L. piscicola, and the two unidentified poultry taxa be classified in a new genus, Carnobacterium, as Carnobacterium divergens comb. nov., Carnobacterium piscicola comb. nov., Carnobacterium gallinarum sp. nov., and Carnobacterium mobile sp. nov. The type strains of C. gallinarum and C. mobile are NCFB 2766T and NCFB 2765T, respectively.

~ Thornley (22) reported the isolation of some gram- DNA studies. Cells for deoxyribonucleic acid (DNA) stud- positive, catalase-negative, nonsporeforming rods from ies were obtained by cultivation in 1 liter of YGPB medium chicken meat maintained at low temperature. These bacteria at 30°C from a 1% inoculum, using a 16-h culture. On resembled lactobacilli in many respects but differed in their reaching early stationary phase, the cells were chilled in ice inability to grow on acetate media. A subsequent, more water and harvested by centrifugation. Preparation of DNA comprehensive study indicated that these strains formed followed a modification (3) of the method of Garvie (4). DNA three groups, none of which at that time corresponded to any base composition was estimated by thermal denaturation in Lactobacillus species or any other recognized taxon (23). A standard saline citrate as described by Garvie (9,using recent numerical phenetic study that included the “atypical DNA from Leuconostoc mesenteroides NCDO 768 (39.3 lactobacilli” described by Thornley and Sharpe confirmed mol% guanine plus cytosine [G+C]) and Escherichia coli the presence of three major groups (S. Ferusu and D. Jones, NCDO 1984 (K-12) (51.5 mol% G+C) as standards. DNA- J. Gen. Microbiol., in press). One of these was shown to DNA hybridizations were performed by using the membrane correspond to Brochothrix thermosphacta, whereas the tax- filter technique described previously (6). onomic position of the other two groups remained unre- Fatty acid analyses. Fatty acids were extracted from dry solved. In recent years there has been an increasing number cells by acid methanolysis (13), and purified methyl esters of reports of unusual lactobacilli from vacuum-packed meat were examined by gas-liquid chromatography, using an Erba that are also unable to grow on acetate agar (8,12,18). These Science Fractovap series 4160 flame ionization chromato- studies have resulted in the establishment of two new graph. Analyses were performed on a 50-m fused-silica species, Lactobacillus carnis (19) and Lactobacillus capillary column with OV-1 as the stationary phase at 200”C, divergens (12). Unfortunately, these two species were not using helium as carrier gas. The identity of individual esters included in the numerical study of Ferusu and Jones, and as was established by comparison of the retention times with far as we are aware there has been no comparative study of those of standard straight-chain saturated, monounsatu- the poultry isolates and vacuum-packed meat strains. There- rated, cyclopropane ring, and methyl-branched esters fore, in the present study the biochemical and chemical (BAME Mix [4-54361; Field Analytical). The identity of characteristics of the atypical lactobacilli described by unsaturated and cyclopropane ring acids was confirmed by Thornley and Sharpe (23) have been determined in an catalytic hydrogenation (2). attempt to clarify their relationship, if any, to the species L. Murein analyses. Purified walls were prepared from ca. 250 carnis and L. divergens. The recently described salmonid mg of dry cells as described by Schleifer and Kandler (15,- fish pathogen Lactobacillus piscicola was also included in 16). The amino acid composition of complete hydrolysates the study because this species shares many properties with was determined by using an automatic amino acid analyser the atypical lactobacilli from poultry and meat (9). (model 4101; LKB Instruments). The isomers of diaminopi- melic acid were separated by thin-layer chromatography as MATERIALS AND METHODS described by Harper and Davis (7). Biochemical and physiological tests. Biochemical tests were Cultures and cultivation. The test strains used are listed in performed by using the API 10E and API 50CH systems Table 1. Cells for peptidoglycan and lipid analyses were (Analytab Products) according to the manufacturer’s in- grown in YGPB broth (5) at 30°C for 3 days. Cultures were structions. YGA medium (nutrient broth no. 2 [Oxoid Ltd.], checked for purity, harvested by centrifugation, washed 25 g; yeast extract [Oxoid], 3 g; glucose, 5 g; agar no. 3 with distilled water, and freeze-dried. [Oxoid], 15 g; water, 1 liter; pH 6.8) was used for test cultures. These cultures were incubated at 25”C, and API * Corresponding author. STREP basal medium was used for carbohydrate tests.

310 VOL. 37, 1987 CARNOBACTERIUM GEN. NOV. 311

Motility was tested by stab inoculation into YGPB medium acetate agar (8, 12, 18, 19). Holzapfel and Gerber (12) modified to contain 0.1% glucose, 0.1% lactose, and 0.2% isolated some atypical heterofermentative bacteria and con- (wthol) agar (Oxoid no. 3). Growth temperatures were cluded that these strains represented a new species of the determined in YGPB medium supplemented with 1% lac- genus Lactobacillus, for which the name L. divergens was tose; readings were made at 48 h, 7 days, and 14 days. Gas proposed. Shaw and Harding (18) isolated some atypical production from glucose was tested in Gibson milk agar and “streptobacteria” from vacuum-packed meat which pos- MRS broth as modified for the arginine test (MRS supple- sessed many properties in common with the isolates de- mented with 0.3% L-arginine monohydrochloride and with scribed by Holzapfel and Gerber (12). In a subsequent study, sodium citrate replacing ammonium citrate) . the isolates described by Shaw and Harding were found to consist of two genetic groups, one of which corresponded to RESULTS L. divergens (19). The second group was considered to represent a new species, for which the name L. carnis was The G+C contents of the DNAs of the test strains are proposed (19). It is also now well established that large shown in Table 1. Strains of Thornley and Sharpe groups 1, numbers of unusual streptobacteria develop on poultry 2, and 3 possessed relatively low G+C ranges of 34.3 to 37.4, stored at low temperatures (1, 17). In the present study, we 34.2 to 36.4, and 35.5 to 37.2 mol%, respectively. These examined the biochemical and genetic inter-relationships of values are similar to those found for the type strains of the the poultry isolates of Thornley and Sharpe, meat strains recently described species L. carnis (35.5 mol%), L. designated L. carnis and L. divergens, and other lactic acid divergens (35.5 mol%), and L. piscicola (34.5 mol%) (Table bacteria. Thornley and Sharpe group 1 strains were found to 1). possess high DNA base sequence relatedness to B. The results of DNA-DNA hybridization experiments are thermosphacta NCDO 1679T consistent with the results of also shown in Table 1. All Thornley and Sharpe group 1 the numerical phenetic study of Ferusu and Jones (in press). strains exhibited high homologies (ca. 60 to 100%) with B. On the basis of biochemical and genetic studies, Thornley thermosphacta NCDO 1676T and negligible relatedness to and Sharpe group 2 strains were found to be heterogeneous. group 2 and 3 strains and the other reference Lactobacillus DNA-DNA hybridizations demonstrated that 16 of these species examined. Thornley and Sharpe group 2 strains strains could be assigned to L. divergens, whereas 15 strains formed three groups on the basis of DNA-DNA hybridiza- were highly related to the type strains of L. carnis and L. tions. Fifteen strains formed a tight group which exhibited piscicola. The recovery of L. carnis and L. piscicola in the high DNA relatedness with L. piscicola NCDO 2762T. L. same DNA homology group was unexpected. Biochemical carnis NCDO 2764T was also a member of this group (Table and chemical data, however, reinforce this relatedness and 1). Sixteen other strains formed a distinct homology group indicate that these species should be reduced to synonymy with high DNA relatedness to L. divergens NCDO 2763T. (the specific epithet piscicola [9] has priority over carnis [19, Two strains (MT44 and 45) were distinct from the above 201). The biochemical and nucleic acid data showed that the homology groups and showed low relatedness with the other remaining group 2 strains were unrelated to the other species reference species examined. Strains of Thornley and Sharpe examined and warrant separate specific status. On the basis group 3 also formed a distinct group on the basis of DNA- of DNA-DNA hybridizations, Thornley and Sharpe group 3 DNA hybridizations and were unrelated to all of the other strains formed a tight group unrelated to other Thornley and strains examined (Table 1). Sharpe isolates and reference Lactobacillus species exam- Chromatographic analysis of the murein hydrolysates re- ined. Group 3 strains were recently found to be a distinct vealed the presence of meso-diaminopimelic acid in all of the taxon in the numerical phenetic study of Ferusu and Jones Thornley and Sharpe strains. Alanine and glutamic acid were (in press). The nucleic acid findings, together with the also detected, which is indicative of a directly cross-linked phenetic distinctiveness of this group, indicate that these murein based on meso-diaminopimelic acid (variation Aly). strains are worthy of separate species status. The cellular fatty acid composition of the Thornley and L. divergens, L. piscicola, L. carnis, and Thornley and Sharpe group 2 and 3 strains together with that of the type Sharpe group 2 and 3 strains possess a number of properties strains of L. carnis, L. divergens, and L. piscicola are shown which are quite different from those of normal lactobacilli. in Table 2. All of the strains possessed predominantly For example, L. divergens, L. carnis, and Thornley and straight-chain saturated and monounsaturated types. Strains Sharpe strains fail to grow on acetate agar broth (14) with a high DNA relatedness to L. divergens NCDO 2763T commonly used for the selective isolation of members of this differed from the other Thornley and Sharpe strains, L. genus. They also differ from traditional heterofermentative piscicola and L. carnis, in possessing substantial levels (ca. Lactobacillus species in producing most of their lactate as 20%) of a CI9 cyclopropane-ring-containingacid. Gas chro- the L-(+)-isomer (12, 19). The cellular fatty acids of these matographic analysis demonstrated that the latter corre- taxa also differ markedly from those reported for other sponded to 9,10-methylenoctadecanoicacid. lactobacilli. It is now established that Lactobacillus species The results of biochemical and physiological tests are possess predominantly straight-chain saturated and mono- shown in Table 3. Strains of Thornley and Sharpe groups 1to unsaturated (and sometimes cyclopropane derivative) acids. 3 were readily separated from each other by the API 50CH All reports to date indicate that the predominant C18:1 isomer system. Strains belonging to Thornley and Sharpe group 2 synthesized is cis-vaccenic acid (A11,12) (10, 11, 24, 25, 27), fell into three groups, two of which were phenotypically and if a C19 cyclopropane ring acid is produced, this is similar to L. divergens and L. piscicolall. carnis. invariably lactobacillic acid (11,12-methylenoctadecanoate) (21,24,25,26,27). L. divergens, L. piscicola, L. carnis, and DISCUSSION Thornley and Sharpe group 2 and 3 strains also synthesize predominantly straight-chain saturated and monounsatur- Over the past few years there have been an increasing ated acids. The major Clgrlisomer produced by these taxa, number of reports of the presence of unusual lactobacilli however, is oleic acid (A9,lO), indicative of quite different from vacuum-packed meat which are unable to grow on unsaturated fatty acid synthase pathway. L. divergens also 312 COLLINS ET AL. INT. J. SYST.BACTEFUOL.

TABLE 1. DNA base composition and DNA-DNA hybridization [‘HIDNA from: Strain” Mol % G+C B. thermosphacta NCDO 1676= C. piscicolu C. divergens C. gallinurum C. mobile MT44 OD MT39L OD Opb St MT21 Op MT59Op MT47 Op MT53 Op MT7 37.4 108‘‘ 70 MT8 37.1 107 79 MT9 36.5 73 73 4 8 5 MTlO 37.0 93 66 7 10 10 MTll 36.9 70 61 8 7 4 MT12 36.5 70 70 27 15 9 MT13 37.3 93 65 10 14 10 MT14 37.3 86 77 13 12 MT15 37.4 91 102 MT16L 37.3 106 82 MT16S 37.0 96 75 MT17 37.3 93 74 MT18 34.3 67 47 15 6 MT19 36.4 66 64 MT20 37.1 108 65 MT33 37.3 79 62 c1 37.3 89 80 c2 36.7 101 62 c3 35.8 84 74 c4 36.4 69 64 NCDO 1676T 37.2 100 100 3 13 9 6 12 6 MT1 35.3 12 75 80 30 18 12 MT2 35.2 87 129 20 MT3 35.4 114 128 30 MT5 36.4 93 118 33 MT2 1 35.0 15 100 80 27 21 15 14 MT27 34.3 87 106 38 13 MT29 34.8 78 99 MT30 35.2 91 109 MT31 34.2 81 84 27 22 MT32 34.8 80 102 28 17 MT35 34.6 94 116 28 MT54 34.0 88 103 34 MT57 35.4 74 114 MT58 35.1 93 111 MT59 34.4 72 100 28 18 40 NCDO 1230 34.9 12 4 92 102 37 39 23 L. curnis (NCDO 2764T) 35.5 12 5 94 127 46 37 27 L. piscicolu (NCDO 2762T) 34.5 15 4 115 100 37 38 28 MT4 35.8 32 111 99 MT6 35.8 33 39 83 89 MT22 36.2 36 84 73 MT23 35.0 18 17 38 79 72 25 MT26 35.6 32 81 78 MT28 35.7 23 91 81 MT46 35.0 22 51 107 73 MT47 35.3 28 33 100 73 35 MT48 34.9 22 35 89 66 MT49 35.5 29 90 93 MT5O 35.9 23 84 101 MT5 1 35.6 29 75 85 MT52 35.3 19 80 84 MT53 35.9 20 38 84 100 20 L. divergens (NCDO 2763T) 35.5 14 4 43 39 89 94 27 MT44 34.3 12 5 13 44 33 18 100 21 MT45 34.4 11 5 17 45 30 23 95 MT36 37.2 87 MT37L 37.0 90 MT37S 36.8 103 MT38 36.5 103 MT39L 36.7 17 14 22 21 18 20 100 MT39S 37.2 73 MT40 35.8 96 MT41 35.5 89 Continued on following page VOL. 37, 1987 CARNOBACTERIUM GEN. NOV. 313

TABLE 1-Continued ['HIDNA from:

Strain" Mol % G+C B. thermosphactci NCDO 1676T C. piscicolu C. divergens C. gallinarum C. mobile MT44 Op MT39L Op Op' St MT21 Op MT59Op MT47 Op MT53 Op MT42L 36.6 100 MT42S 36.9 103 MT43L 36.3 87 MT43S 36.2 73 DA2 42.4 12 5 26 21 25 21 20 22 L. murinus (NCDO 2175) 40.3 9 4 13 10 12 10 13 "L. intestinalis" (NCDO 2176) 36.6 8 4 13 14 8 9 12 L. alimentarius (NCDO 2329T) 32.9 10 4 16 16 11 15 13 L. bavaricus (NCDO 2588T) 38.1 11 4 23 20 17 21 22 L. farciminis (NCDO 2380T) 35.6 7 4 15 11 9 8 8 ' MT strains are from poultry (see reference 22): 7 to 20 and 33, Thornley and Sharpe group 1; 1 to 6,21 to 23. 26 to 32, 35, and 44 to 59, Thornley and Sharpe group 2; 36 to 43, Thornley and Sharpe group 3. Strains C1 and C2 were isolated from meat; C3 and C4 were isolated from lamb; NCDO 1230 was isolated from hu- man plasma (ca. 1955); and DA2 was isolated from pate. Op, Optimum conditions; St, stringent conditions. Values of >60% are in boldface type. synthesizes the corresponding cyclopropane ring acid basis of DNA-DNA hybridization (ca. 12% homology). dihydrosterculic acid (9,lO-methylenoctadecanoate) (Table Furthermore , the presence of respiratory quinones (MK-7) 4). The above differences, therefore, lead us to believe that and predominantly iso- and anteiso-methyl-branched fatty L. divergens, L. piscicola (including L. carnis), and acids in B. thermosphacta indicate that these taxa are not Thornley and Sharpe group 2 and 3 strains are not true members of the same family. We therefore formally propose lactobacilli and warrant a separate taxon. L. divergens, L. that L. divergens and L. piscicola (synonym, L. carnis) be piscicola, and Thornley and Sharpe group 2 and 3 strains reclassified in a new genus, Carnobacterium, as Carno- also exhibit low relatedness to B. thermosphacta on the bacterium divergens comb. nov. and Carnobacterium

TABLE 2. Cellular fatty acid composition"

Species % of total assignment of strains c14 I c14 1 C14 0 Clh 1 A9 CIh I clh0 CIS 1 A9 CIR 1 CIR 0 ccyclo-19 A9 C. divergens NCDO 2763T 1.5 2.5 10.5 11.5 1.5 20.0 28.0 2.0 4.0 19.5 MT4 1.o 2 .o 11.0 11.0 1.0 21.0 29.0 1.o 4.5 18.5 MT6 1.5 2.0 11.0 14.5 2.5 19.0 26.0 1.o 3.0 19.5 MT26 1.0 2.0 10.5 11.5 1.5 20.0 28.0 1.o 4.0 20.5 MT46 0.5 1.5 10.0 11.0 1.o 19.5 27.5 1.o 3.5 23.5 MT47 1.o 2.5 10.0 12.0 1.5 20.5 28.5 1.o 3.5 19.5 MT48 1.o 2.5 9.5 12.5 2.0 19.5 29.5 1.o 4.5 18.0 MT49 1.o 1.5 8.5 12.0 2.0 18.5 31.0 0.5 3.5 21.5 MT50 0.5 1.5 10.0 13.5 2.0 19.0 30.0 0.5 3 .O 20.0

C. gallinarum MT44 1.0 2.0 13.0 8.5 1.5 26.5 40.5 1.5 5.5 MT45 0.5 2.0 12.5 8.0 1.o 26.0 42.5 1.5 6.0

C. mobile MT37 0.5 1.0 2.0 25.5 2.5 16.0 48.5 1.5 2.5 MT38 0.5 1.o 2.0 25.5 3 .O 17.0 47.5 1.5 2.0 MT39 0.5 1.0 2.0 27.0 2.5 17.5 46.0 1.5 2.0 MT4 1 0.5 1.0 2.0 28.0 3.0 15.5 46.5 2.0 1.5 MT42 0.5 1.5 2.0 27.5 2.5 16.5 46.0 1.5 2.0

C. piscicola NCDO 2762T 1.o 3.0 10.0 11.0 2.0 17.5 51.5 0.5 3.5 MT2 1.0 3.0 11.0 16.0 2.0 14.0 51.0 0.5 1.5 MT5 0.5 2.5 10.5 12.5 1.5 16.5 53.5 0.5 2.0 MT21 1.o 2.5 11.0 13.0 2.0 16.0 51.5 0.5 2.5 MT58 1.0 2.5 10.5 13.5 2.0 15.5 52.0 0.5 2.5 MT59 1.0 2.0 11.0 12.5 2.0 18.5 49.5 0.5 3.0 NCDO 2764b 1.o 2.5 11.0 12.0 2.0 17.5 50.0 1.o 3.0

a Clb,,, Hexadecanoic acid; Cl8,, octadecenoic acid; Ccyclo-Ip,methylenoctadecanoic acid. ' Formerly the type strain of L. ccirnis. 314 COLLINS ET AL. INT. J. SYST.BACTERIOL.

TABLE 3. Biochemical and physiological properties of test strains Reaction" Thornley and Sharpe group 2' Characteristic Thornley and L. carnis L. divergens L. piscicola sh~~~~~~~3' NCDO 2764T NCDO 2763T NCDO 2762' 54, 57 to 59 Acid produced from:': Amidon 3/16 7/15 112 - - Am ygdalin + + + + + Galactose 6/16 10115 + + + + P-Gentiobiose + + + 10112 + + Gluconate 15/16 + +W - + + Inulin - + - + + + 2-Ketogluconate - - - 4/12 - - Lactose 2/16 10115 + - + + Mannitol - + - + + Melezitose 9/16 14/15 +D + + Melibiose - 7/15 - + + a-Methyl-D-glucoside + + + + a-Methyl-D-mannoside + - - + + D-Tagatose - + 1/12 - - D-Turanose 14/15 +W - + +D D-Xylose +D - - Voges-Proskauer"' + + - + + Motility - + - - Gas production from glucose in arginine- MRS are broth' + + 11/12 + + + Growth at temp ("C): 0 + + + + + 30 + + + + + 37 + + 9/12 + + 40 11/16 - - - - 45 - Number of strains positive/total; +, all strains positive; -, all strains negative; +W, weak positive: +D, positive after 7 days. MT strain numbers. Acid from carbohydrates was determined by using the API 5OCH system; readings were made at 7 days. All strains produced acid from cellobiose, D-fructose, D-glucose, maltose, D-mannose, N-acetylglucosamine, ribose, salicin, sucrose, and trehalose; acid was not produced from adonitol, D-arabinose, L-arabinose, D- arabitol, L-arabitol, dulcitol, erythritol, D-fucose, L-fucose, glycogen, inositol, 5-ketogluconate, D-lyxose, P-methylxyloside, raffinose, L-rhamnose, sorbitol, L- sorbose, xylitol and L-xylose. Tests performed on API 10E system; all strains produced arginine dihydrolase and P-galactosidase; all strains were lysine decarboxylase, tryptophan desaminase, urease, ornithine decarboxylase, indole, and H2S negative. All strains failed to produce gas in Gibson milk agar.

TABLE 4. Characteristics useful in differentiating piscicola comb. nov., respectively. We also consider that Carnobacterium species Thornley and Sharpe group 3 isolates and strains MT44 and Reaction" MT45 (group 2) are worthy of separate species status for Characteristic C. C. C. C. which the names Carnobacterium mobile sp. nov. and divergens gallinarum mobile piscicola Carnobacterium gallinarum sp. nov., respectively, are pro- Acid produced fromb: posed. Characteristics useful in the differentiation of Am ygdalin + + Carnobacterium species are summarized in Table 4. Gluconate +(-I + Description of Carnobacterium gen. nov. Carnobacterium Inulin - - (L. gen. n. carnis, of flesh; Gr. dim. n. bakterion, a small Mannitol - - rod; M.L. neut. n. Carnobacterium,flesh rodlet). Nonspore- Melezitose V + forming, straight, slender rods usually occurring singly or as a-Methyl-D-glucoside - + pairs but sometimes as short chains. Cells are gram positive. a-Methyl-D-mannoside - - May or may not be motile. Heterofermentative, producing D-Tagatose - + D-Turanose - + predominantly L-( +)-lactic acid from glucose (12, 19). Gas D-Xylose - + production is variable (dependent on substrate) and fre- Voges-Proskauer + + quently negative. Growth occurs at 10°C; most strains grow Motility - - at 0°C but not at 45°C. No growth in 8% NaC1. Does not 9,lO-Methylenoctadecanoic grow on acetate agar or broth (14). Catalase and benzidine acid' + - negative. Nitrate is not reduced to nitrite. The peptidoglycan '' -(+I, Occasional strain positive; +(-I, occasional strain negative. is of the meso-diaminopimelic acid direct type. Major cellu- ' Reading performed at 7 days. lar fatty acids are of the straight-chain saturated and ' >15% of total cellular fatty acids. monounsaturated types; the C18:1 compound is a A9,10 VOL. 37, 1987 CARNOBACTERIUM GEN. NOV. 315 isomer. Cyclopropane-ring-containingfatty acids may also (midpoint melting temperature [ TJ). Isolated from irradi- be produced. The G+C content of DNA is 33.0-37.2 mol%. ated chicken meat. Found in vacuum-packaged meat and related products The type strain is NCFB 2765T (MT37L). The description stored at low temperatures. Also isolated from salmonid fish of the type strain corresponds to that of the species, except (causative agent of lactobacillosis) but may also occur in that growth occurs at 37”C, acid is produced from P- nonsalmonid species. gentiobiose, and acid is not produced from D-tagatose. The type species of Carnobacterium is Carnobacterium Description of Carnobacterium gallinarum sp. nov. Carno- divergens. bacterium gallinarum (gal.li.na’rum. L. fem. n. gallina, a Description of Carnobacterium divergens (Holzapfel and hen; L. fem. gen. pl. n. gallinarum, of hens). Gram-positive, Gerber 1983) comb. nov. The full description of the type nonsporeforming, straight, slender rods which occur singly species C. divergens is as given by Holzapfel and Gerber or in short chains. Cells are nonmotile. Colonies are white, (12). The description, however, should be extended as convex, shiny, and circular. Growth occurs at 0 and 35°C but follows. Major cellular fatty acids are of the straight-chain not at 40°C. Facultatively anaerobic. L-( +)-Lactic acid is saturated, monounsaturated, and cyclopropane ring types produced. No gas is produced from glucose. Acid is pro- with tetradecanoic, hexadecanoic, hexadecenoic, 9,lO- duced from amygdalin, cellobiose, D-fructose, galactose, octadecenoic, and 9,lO-methylenoctadecanoic acids pre- P-gentiobiose, gluconate, D-glucose, lactose, maltose, D- dominating. The G+C content of DNA ranges from 33.0 to mannose, melezitose, a-methyl-D-glucoside, N-acetylgluc- 36.4 mol%. osamine, D-ribose, salicin, sucrose, D-tagatose, D-turanose, The biochemical characteristics of the type strain are trehalose, and D-xylose. Acid is not produced from adonitol, given in Table 3. Acid is produced from gluconate and D-arabinose, L-arabinose, D-arabitol, L-arabitol, dulcitol, melezitose. Acid is not produced from amidon, galactose, or erythritol, D-fucose, L-fucose, glycogen, inositol, inulin, lactose. Growth occurs at 40°C. 2-ketogluconate, 5-ketogluconate, D-lyxose, mannitol, meli- Description of Carnobacterium piscicola (Hiu, Holt, biose, a-methyl-D-mannoside, P-methyl+-xyloside, raf- Sriranganathan, Seidler, and Fryer 1984) comb. nov. The full finose, L-rhamnose, sorbitol, L-sorbose, xylitol, and L- description of C. piscicola is as given by Hiu et al. (9) but xylose. Acid from amidon variable. All strains hydrolyze should be extended and emended as follows. Under certain esculin and are arginine dihydrolase and P-galactosidase conditions exhibits heterofermentative properties. Gas is positive. All strains are lysine decarboxylase, ornithine produced from glucose in arginine-MRS broth. Major cellu- decarboxylase, tryptophan desaminase, and urease nega- lar fatty acids are of the straight-chain saturated and tive. Indole and H2S are not produced. Voges-Proskauer monounsaturated types with tetradecanoic, hexadecanoic, positive. hexadecenoic, and 9,lO-octadecenoic acids predominating. The cell wall peptidoglycan contains meso-diaminopimelic The G+C content of DNA ranges from 33.7 to 36.4 mol%. acid. The cellular fatty acids are of the straight-chain satu- The biochemical characteristics of the type strain are rated and monounsaturated types with tetradecanoic, given in Table 3. Acid is produced from galactose, lactose, hexadecanoic, and 9,lO-octadecenoic acids predominating. melibiose, melezitose, and D-turanose. Acid is not produced The G+C content of DNA ranges from 34.3 to 36.4 mol% from amidon. (Tm).Isolated from ice slush from around chicken carcasses. Description of Carnobacterium mobile sp. nov. Carno- The type strain is NCFB 2766T (MT44). The description of bacterium mobile (mo’bi.le. L. neut. adj. mobile, movable or the type strain corresponds to that of the species, except that motile). Gram-positive, nonsporeforming, straight, slender acid is not produced from amidon. rods which occur singly or in short chains. Cells are motile. Colonies are white, convex, shiny, and circular. Growth LITERATURE CITED occurs at 0 and 35°C (some strains do not grow at 37°C) but not at 40°C. Facultatively anaerobic. L-(+)-Lactic acid is 1. Barnes, E. M. 1976. Microbiological problems of poultry at refrigerator temperatures-a review. J. Sci. Food Agric. 27: produced heterofermentatively. Gas production from glu- 777-782. cose is observed for most strains in arginine-MRS broth. 2. Brian, B. L., and E. W. Gardner. 1968. A simple procedure for Acid is produced from cellobiose, D-fructose, galactose, detecting the presence of cyclopropane fatty acids in bacterial D-glucose, inulin, maltose, D-mannose, N-acetylglucos- lipids. Appl. Microbiol. 16549-552. amine, D-ribose, salicin, sucrose, and trehalose. Acid is not 3. Farrow, J. A. E., D. Jones, B. A. Phillips, and M. D. Collins. produced from adonitol, amidon, amygdalin, D-arabinose, 1983. Taxonomic studies on some group D streptococci. J. Gen. L-arabinose, D-arabitol, L-arabitol, dulcitol, erythritol, D- Microbiol. 129: 1423-1432. fucose, L-fucose, glycogen, gluconate, inositol, 5- 4. Garvie, E. I. 1976. Hybridization between the deoxyribonucleic ketogluconate, lactose, D-lyxose, mannitol, melezitose, acids of some strains of heterofermentative . Int. J. Syst. Bacteriol. 26:116-122. melibiose, a-methyl-D-glucoside, a-methy1-D-mannoside, P- 5. Garvie, E. I. 1978. 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