The name Bacillus azotoformans (ex Pichinoty, de Barjac, Mandel, Greenway, and Garcia 1976) is revived. Strains of this species are motile and produce terminal or subterminal oval endospores in swollen sporangia. Nitrate, nitrite, and nitrous oxide are denitrified with production of nitrogen. The guanine-pluscytosine content of the deoxyribonucleic acid of this organism is 39.8 2 1.2 mol% (mean 2 standard deviation). The type strain is ATCC 29788 (= CCM 2849 = CIP R925).The name Bacillus azotoformans Pichinoty , de Barjac, Mandel, Greenway, and Garcia 1976 does not appear on the Approved Lists of Bacterial Names (9), although it was effectively published (6) and a type strain was designated (7). The purpose of this report is to revive the name Bacillus azotoformans and to effect its valid publication. Although this species has been described previously (6), we provide a formal description, including formerly unpublished fatty acid and phospholipid compositions.Fatty acid and phospholipid composition studies were done by using strains 1 , 13, and 30 (7). These strains were grown for 24 h at 32°C in yeast extract-salts liquid medium; this medium was made by combining 4 g of yeast extract (Institut Pasteur, Lille, France), 3.575 g of Na2HP04 . 12H20, 0.98 g of KH2P04, 0.03 g of MgS04.7H20, 0.5 g of NH4C1, 0.2 ml of a solution containing calcium and various heavy metals chelated with ethylenediaminetetraacetate (8), and distilled water to a volume of 1 liter. Under these conditions, poly-p-hydroxybutyrate is not synthesized. Cells were collected by centrifugation, washed with distilled water, and freeze-dried. The total free lipids were extracted with chloroform-methanol (2:1, vol/vol). After evaporation of the solvents in a vacuum, the residue was extracted with ether. Silica Gel G plates (E. Merck AG, Darmstadt, Germany) were spotted with approximately 10 p, g of each sample and then developed in ascending fashion by using a solution containing petroleum ether (bp 50°C) and diethyl ether (4:1, vol/vol). Staining of the developed plates with Rhodamine B revealed no more than traces of triglycerides. 660Further development with chloroform-methanol-water (65:25:4, vol/vol) and successive staining were performed with ninhydrin and Dittmer reagents. For fatty acid gas chromatography, the lipids were saponified in 5% (wt/vol) KOH that was dissolved in 33% (vol/vol) aqueous methanol at 100°C for 90 min. Fatty acids were methylated by reacting them with diazomethane in diethyl ether and then injected into a 3-m glass column containing 1% OV-1 on Chromosorb W.

We have studied 43 strains of the species Alcaligenes dentrificans, A. odorans, and A. faecalis. Twenty-five of them were isolated by enrichment culture on minimal medium containing an organic acid (L-malate, succinate, tartrate, adipate, or itaconate) and N2O as a respiratory electron acceptor. These constitute a single phenon with the A. dentrificans strain type and 9 other strains isolated from clinical specimens. However, strain 4 differs from the other 34 strains in 12 nutritional characters, in its ability to effect a meta cleavage of diphenols, and by the absence of tetrathionate reductase. The percentages of G + C are the following: strains isolated from soil, 66.4 +/- 1.1; collection strains, 67.0 +/- 1.3. The 5 strains of A. odorans differ from the 34 strains of A. denitrificans (not including strain 4) in their inability to denitrify nitrate and use D-saccharate, adipate, pimelate, suberate, beta-hydroxy-beta-methylglutarate meso-tartrate, azelate, and itaconate. Their percentage of G + C is much lower: 56.1 +/- 0.4. From the nutritional point of view the 3 strains of A. faecalis resemble A. dentrificans. However, they differ from the latter by their inability to grow anaerobically on NO3-, NO2-, N2O, and by a slightly lower percentage of G+ C: 64.3 +/- 0.0. The 43 strains synthesize poly-beta-hydroxybutyric acid. None of them is chemolithotrophic.

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The denitrifying capacity of 15 strains of Bacillus licheniformis was evaluated. In general, N2 production by the cultures on complex media containing NO3- is irregular and quite slow and three of the strains never produce gas. Bacillus licheniformis grows rapidly in anaerobiosis on peptone medium containing NO3- which is reduced to NO2-. None of the strains grow in peptone medium with NO2- or N2O as the respiratory substrate, nor do they grow under an atmosphere of 10% NO-90% N2. Denitrification was studied in cell suspensions using gas chromatography. N2O production from NO3- or NO2- is always weak at best; nitric oxide is reduced to N2O at an appreciable rate. All the strains synthesize nitrate reductase A in anaerobiosis when NO3- is present. In cell extracts, nitrite reductase activity is always negligible or nil with tetramethyl-p-phenylenediamine as an electron donor.

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