J Clin Pathol: first published as 10.1136/jcp.25.11.959 on 1 November 1972. Downloaded from J. clin. Path., 1972, 25, 959-965 Identification and characterization of Moraxella phenylpyruvica J. J. S. SNELL, L. R. HILL, AND S. P. LAPAGE From the National Collection of Type Cultures, Central Public Health Laboratory, London SYNOPSIS Eight strains of Moraxella phenylpyruvica have been isolated from clinical material in the United Kingdom, the first to be reported from this country. They were characterized, together with three strains of M. phenylpyruvica of the National Collection of Type Cultures (NCTC), and com- pared with NCTC strains of eight other Moraxella species. The strains of M. phenylpyruvica formed a homogeneous group which is readily distinguishable from otherMoraxella species. Deamination of phenylalanine is not restricted to M. phenylpyruvica which, however, is urease positive and is stimulated by bile, in contrast to other Moraxella spp. Flamm (1957) described a strain of Moraxella which fluid, nasal swabs, and various lesions) have been he named M. polymorpha, isolated from the cerebro- examined in the National Collection of Type spinal fluid of a child with meningitis. Bovre and Cultures (NCTC), together with the three NCTC Henriksen (1967) examined this strain and found it to strains of this species. During a comparison of these by copyright. agree in a number of characters with eight other strains with eight other Moraxella species, it was Moraxella strains from such clinical sources as the found that the ability to deaminate phenylalanine is urogenital tract, blood cultures, and pus. They not limited to M. phenylpyruvica (Snell and Davey, considered that Flamm's strain belonged to the same 1971). However, M. phenylpyruvica can be distin- species as the other eight strains and suggested that guished from other Moraxella species by several the epithet 'polymorpha' in Moraxella polymorpha other characters as well as deamination of phenyl- was illegitimate and would cause confusion with De alanine. The purpose of the present work is to record Bord's (1939, 1942) Mima polymorpha. The strains the isolation of this little-known organism in the were found to deaminate phenylalanine, a property UK, and to record a number of hitherto undescribed http://jcp.bmj.com/ which was thought to be unusual in the genus characteristics which help to distinguish it from other Moraxella, and B0vre and Henriksen proposed re- Moraxella species. naming this species M. phenylpyruvica (see also Judicial Commission of the International Committee Materials and Methods on Systemic Bacteriology, 1971). Eight strains of M. phenylpyruvica isolated in the ORGANISMS UK from clinical material (urogenital tract, pleural Eleven strains of M. phenylpyruvica were examined on September 26, 2021 by guest. Protected Received for publication 17 August 1972. (Table I): eight field strains sent to the NCIC for Strain No. Original Strain Designation Receivedfrom Source I A152/68 M. P. Jevons, London Vulva 2 A12/70 R. F. Alexander, London Urine 3 A30/70 E. G. Gordon, Worcestershire Pleural fluid 4 A58/70 E. G. Gordon, Worcestershire Vagina 5 A80/70 J. C. Bume, Dartford Nasal swab 6 A81/70 K. B. Rogers, Birmingham Pressure sore 7 A84/70 W. E. Rimington, Doncaster Bartholin cyst 8 A100/71 I. R. Poulter, Warwick Leg ulcer NCTC 10526 2863 American Type Culture Collection, ATCC 23333 Blood NCTC 10750 5542 American Type Culture Collection, ATCC 23334 Blood NCTC 10752 - American Type Culture Collection, ATCC 17958 Cerebrospinal fluid Table I Strains of Moraxella phenylpyruvica examined in the present work 959 J Clin Pathol: first published as 10.1136/jcp.25.11.959 on 1 November 1972. Downloaded from 960 J. J. S. Snell, L. R. Hill, anid S. P. Lapage identification; three NCTC strains including the Strain 7 neotype of M. phenylpyruvica (NCTC 10526) and A 26-year-old woman had for three years a Bartholin Flamm's strain of M. polymorpha (NCTC 10752). cyst which became infected and from which this Strains of eight other Moraxella species were strain was isolated. After excision of the cyst, she examined for comparison and are listed in Table II. fully recovered. Strain 8 Name Number A man with a severe ankle injury developed a leg M. bovis NCTC 8561, 9425, 9426, A52/69' ulcer after seven weeks in plaster. The strain was M. kingii NCTC 10526, 10746 isolated from the ulcer on two occasions before and M. lacunata NCTC 7985, 10747, 10748 M. liquefaciens NCTC 7911, 10358, 10359 during treatment with first fucidin, then ampicillin, M. nonliquefaciens NCTC 7784, 10464, A33/67, A9/69 and finally ultraviolet light. M. osloensis NCTC 10465, 10749, 10755, 10756 M. saccharolytica2 NCTC 10753 M. sp.3 NCTC 10717 BACTERIOLOGICAL INVESTIGATIONS The media and methods used were mainly those Table It Strains of other Moraxella spp. examined in described in Cowan and Steel (I 965). The media were the present work supplemented where indicated with horse blood or 'Strains with designations prefixed by A are field strains identified serum. The microscopic appearance was examined in the NCTC. after 24 hours' growth at 37°C on 5°O (v/v) blood 'Described by Flamm (1956). 3Described by van Bijsterveld (1970. 1971). nutrient agar. Tests for motility were carried out by the hanging drop method from overnight cultures in 100% (v/v) serum nutrient broth. The colonial CLINICAL DETAILS OF THE FIELD STRAINS appearance was observed after 24 and 48 hours' growth at 37°C on 5% (v/v) blood nutrient agar. Strain 1 Growth at 5°C, 22°C, and 42°C was tested for on A 4-year-old girl, admitted to hospital for investi- blood agar and anaerobic growth on blood agar by copyright. at gation of enuresis, had a mild vulvo-vaginitis. The 37°C in an atmosphere of 9000 (v/v) hydrogen and vaginitis cleared spontaneously. 100% (v/v) carbon dioxide. The strains were tested for growth on nutrient Strain 2 agar, on MacConkey agar, on 0/F medium (Hugh A primigravida with preeclamptic toxaemia was and Leifson, 1953), solidified with 1 % agar, and in found to have excess albumin in the urine. The strain Koser's and Christensen's citrate media. Growth was isolated on one occasion only and a normal with 0.50% (w/v) f-hydroxybutyric acid as a sole delivery took place two weeks later. carbon source was detected on the basal medium of Owens and Keddie (1968). If growth was obtained, http://jcp.bmj.com/ Strain 3 the bacteria were stained with Sudan Black B to test This strain was isolated from pleural fluid of a 67- for the presence of poly-f/-hydroxybutyrate inclusion year-old man, admitted to hospital with diffuse granules. Those strains failing to grow on this pulmonary fibrosis. medium were grown on 10% (v/v) serum nutrient agar containing /3-hydroxybutyric acid and then Strain 4 stained as above. The effect of the following sub- This strain was present in large numbers in a vaginal stances or additives on growth was recorded: 10% on September 26, 2021 by guest. Protected discharge. The normal flora was very scanty. (v/v) serum in nutrient agar; brilliant green in- corporated in blood agar at a concentration of 1 in Strain 5 500,000 (w/v); ox bile in concentrations of 1000 A 14-month-old girl with gastroenteritis yielded this (w/v) and 40% (w/v) and sodium taurocholate in strain from a nasal swab. The gastroenteritis cleared concentrations of 0.5 % (w/v) and 2 % (w/v) in- up rapidly and spontaneously within two days. corporated as ditches in blood agar plates. The strains of M. phenylpyruvica were also tested for Strain 6 stimulation of growth by stearate, oleate, Tween 80, An 11-year-old boy, born with a myelocoele and butyrate, propionate, valerate, acetate, pelargonate, spina bifida, had both faecal and urinary incon- and lecithin, each at a concentration of 01 0% (w/v) tinence. A pressure sore developed in which this in blood agar. strain was present in large numbers together with Minimum inhibitory concentrations (MICs) of Proteus sp. and non-haemolytic and viridans type penicillin were determined by subculture on blood streptococci. agar containing the antibiotic in a range of concen- J Clin Pathol: first published as 10.1136/jcp.25.11.959 on 1 November 1972. Downloaded from Identification and characterization of Moraxella phenylpyruvica 961 trations between 0-0001 and 100 IU/ml. The Oxford were made in either saline-phosphate buffer (SP: strain of Staphylococcus aureus (NCTC 6571) was 0-1 M NaCl + 0-01 M phosphate, pH 7-0) or used as a control and was inhibited at a concentra- standard saline-citrate buffer (SSC: 0-15 M NaCl + tion of0-05 IU penicillin/ml. Tests for the production 0-015 M sodium citrate, pH 7-0). The equations of penicillinase were made by the method of Parker relating Tm to the percentage of guanine + cytosine/ and Lapage (1957), using two strains of Staphylo- total bases (% GC) for these two buffers were: for coccus aureus as controls (NCTC 4136, positive SP buffer: % GC = 2-352 Tm-153 49 (Owen, Hill, control and NCTC 10443, negative control). and Lapage, 1969), and for SSC buffer: % GC = Acid production from glucose was recorded in the 2 44 Tm-169 0 (Marmur and Doty, 1962). O/F medium of Hugh and Leifson (1953), both with and without the addition of 10% (v/v) serum, and Results also on 10% (v/v) serum nutrient agar slopes containing 1 % (w/v) glucose and 0 01 % (w/v) phenol Microscopically, the 11 strains of M. phenylpyruvica red. Catalase production was tested on cultures consisted of non-motile ovals and rods, measuring grown on 10% (v/v) serum nutrient agar, and 0-7p x 1 5-20,u, which were Gram negative, production of oxidase was detected by Kovacs' although all strains showed some cells which retained (1956) method using tetramethyl-p-phenylenedia- the crystal violet dye as intracellular granules.