Biochemical characteristics and fatty acid composition of Gilardi rod group 1 bacteria.

Fifteen strains of eugonic, nonoxidative, gram-negative rods isolated primarily from human wounds of the extremities and blood formed a distinct group which was designated Gilardi rod group 1. The phenotypic characteristics of Gilardi rod group 1 were most similar to those of CDC group M-5, with the major difference that nitrite reduction was observed with CDC group M-5. All 15 strains of Gilardi rod group 1 possessed a distinct fatty acid profile which was characterized by large amounts (> 15%) of cis-vaccenic (18:1 omega 7c), palmitic (16:0), myristic (14:0), and lactobacillic (19:0 cyc11,12) acids and moderate amounts (3 to 5%) of lauric (12:0), 3-hydroxylauric (3-OH-12:0), and palmitoleic (16:1 omega 7c) acids. This fatty acid profile is unique compared with the profiles of CDC group M-5 and other bacteria we have tested and is useful for the rapid identification of Gilardi rod group 1 isolates.

Chemical characterization of clinical isolates which are similar to CDC group DF-3 bacteria.

Six clinical isolates, taken from blood or wounds, that had biochemical characteristics most similar to Centers for Disease Control group DF-3 bacteria were examined for cellular fatty acid composition and isoprenoid quinone content to evaluate their chemical relatedness to known bacterial species and groups. The fatty acids were liberated from whole cells by base hydrolysis, methylated, and analyzed by capillary gas-liquid chromatography. The isoprenoid quinones were extracted from lyophilized whole cells and analyzed by reverse-phase high-performance liquid chromatography. All six strains, which were designated group DF-3-like, possessed a distinct fatty acid profile that was characterized by large amounts (greater than 20%) of 13-methyltetradecanoate (i-C15:0) and 12-methyltetradecanoate (a-C15:0), moderate amounts of saturated branched-chain 13-carbon acids (i-C13:0 and a-C13:0) and hexadecanoate (n-C16:0), and small to moderate amounts of both branched- and straight-chain hydroxy acids (i-3-OH-C15:0, 3-OH-C16:0, i-3-OH-C17:0, and 2-OH-C17:0). This fatty acid profile was unique compared with the profiles of group DF-3 and other bacteria we have previously tested and is useful for the rapid identification of group DF-3-like isolates. The isoprenoid quinone content of four group DF-3-like strains was similar, with ubiquinone-9 (Q-9) and Q-10 as their major quinones, while the other two group DF-3-like strains contained Q-7 as their major quinones, with smaller amounts of Q-8 and Q-9.

Regional variations in the "adenine/oxypurine" pool of the heart in normoxia and oxygen deficiency.

The regional distribution of energy-related metabolites was determined for the normal dog heart by using high performance liquid chromatography to analyse the levels of ATP, ADP, AMP, IMP, adenosine, inosine, hypoxanthine/xanthine, uric acid and NAD in tissue samples from 7 defined areas of the myocardium and 2 of the conduction system, and calculating the overall concentration of these metabolites in each area. From 8 of the 20 hearts used, additional samples were taken and allowed to undergo autolysis in vitro at 37 degrees C for up to 2 h before being analysed in the same manner. The total concentration of the metabolites assayed ("adenine/oxypurine" pool) did differ from one area to another, most notably between ventricles (left = 6.08, right = 5.93, apex = 6.08 microM/g wet tissue weight) and atria (left wall = 4.44, left appendage = 4.12, right wall = 4.34, right appendage = 3.57 microM/g wet tissue weight), but for each area remained essentially constant during the period of autolysis studied.

Neisseria lactamicus sp. n., a lactose-fermenting species resembling Neisseria meningitidis.

The biochemical and serological characteristics of lactose-utilizing strains of Neisseria were determined. These organisms were found in the nasopharynx of man and grew well on Thayer-Martin Selective Medium. They were compared with N. meningitidis to ascertain whether they were variants of this species. Differences between the lactose-using strains and the recognized species of Neisseria were considered significant enough to warrant designation of a new species, Neisseria lactamicus. This group has not been widely recognized as being separate from N. meningitidis; therefore, the normal incidence and clinical significance of these organisms has not been fully established. These organisms are oxidase-positive and positive for beta-D-galactosidase activity; they demonstrate fermentation in King Oxidation-Fermentation Medium; and they produce acid from only glucose, lactose, and maltose, of the 27 substrates incorporated in Cystine Trypticase Agar. Individual strains vary in their ability to grow on Nutrient Agar at both 25 and 37 C and in their pigmentation on Loeffler Medium. Results indicated that these organisms are serologically distinct from the N. meningitidis serogroups. Only 34 of 116 strains of N. lactamicus were smooth and could be tested by slide agglutination. None of the 34 could be grouped as N. meningitidis group A, B, C, D, X, Y, or Z. Thirty-one of these strains could, however, be specifically grouped with antisera prepared with N. lactamicus strains. Cross absorptions confirmed that N. lactamicus is serologically distinguishable from N. meningitidis.

Serological studies of ungroupable Neisseria meningitidis.

Verification that Slaterus' Neisseria meningitidis serotypes X, Y, and Z are groups distinct from each other and from groups A, B, C, and D was made by use of the tube agglutination test on absorbed and unabsorbed antisera. A significant number of meningococcal strains in this country, which could not be classified serologically with standard antisera prepared to Branham's neotype A, B, C, and D strains, were grouped specifically with antisera prepared to the Slaterus types. The strains grouped as X, Y, and Z were from various geographical areas of the United States and were isolated from both carriers and cases. Over a 2-year period, the cultures tested ranged in predominance in descending order as follows: group B, C, Y, X, Z, A, and D. It is recommended that Slaterus' types should be considered as standard groups and follow in alphabetical order with the standard A, B, C, and D groups; i.e., X would be designated as group E, Y as group F, and Z as group G. It was observed that false-grouping cross-reactions could be greatly reduced by reconstituting the lyophilized grouping antisera in 50% glycerol-water. Of 99 cultures which could not be specifically grouped with antisera reconstituted in distilled water, 19 were specifically grouped with antisera reconstituted in 50% glycerol-water.

Fatty acid composition of Neisseria species as determined by gas chromatography.

Duplicate cultures of 53 strains representing 9 species of Neisseria were analyzed by gas-liquid chromatography for cellular fatty acids. N. sicca, N. mucosa, N. flava, N. flavescens, N. perflava, N. subflava, and the several serotypes of N. meningitidis examined were found to comprise a fairly homogeneous group on the basis of the percentages of individual fatty acids present. Lactose-fermenting Neisseria also were in this group. N. catarrhalis, however, contained decanoic acid in addition to the acids occurring in the other species. Moreover, the 18 C-saturated and monoenoic acids together constituted 36% of the total fatty acid composition for N. catarrhalis, while the comparable mean value for the other species was less than 13%.

An unclassified gram-negative rod isolated from the pharynx on Thayer-Martin medium (selective agar).

An oxidase-positive, small gram-negative rod was isolated on Thayer-Martin medium (TM) inoculated with pharyngeal swabs obtained during surveys to detect Neisseria carriers. In one survey, this organism was isolated from 48% of the subjects, and 50 or more colonies were present on the majority of the primary isolation plates. Other characteristics of the organism, which has been given the provisional designation "TM-1," include: delayed production (2 to 10 days) of acid from glucose, formation of gas during nitrate reduction, and the frequent formation of "pits" in the agar surface. On TM, nonpitting colonies of TM-1 are morphologically similar to colonies of Neisseria gonorrhoeae and N. lactamica. Comparison of the characteristics of TM-1 strains with other similar fastidious gram-negative organisms encountered in clinical laboratories indicates that TM-1 is a distinct species. Further studies are required before proper taxonomic placement can be made.

Emended description of Kingella denitrificans (Snell and Lapage 1976): correction of the maltose reaction.

Snell and Lapage proposed that the bacterium designated CDC group TM-1 be named Kingella denitrificans. Their description of the organism indicated that it produced acid from maltose which was in contrast with an earlier report from our laboratory that maltose was not utilized. In an attempt to explain the different findings in the two laboratories, the type strain and two additional strains of K. denitrificans were tested for acid production from maltose in phenol red broth, the rapid fermentation test, fermentation broth with Andrade's indicator, and oxidation-fermentation base. These media were tested without additional supplements and with supplements of horse serum (5% [vol/vol]) or rabbit serum (5% [vol/vol]). The results indicated that K. denitrificans does not produce acid from maltose. It was demonstrated that maltose in media containing serum is converted to glucose from which acid can be produced by K. denitrificans. The supplementation of maltose medium with horse serum by Snell and Lapage apparently had produced a false-positive maltose reaction. The description of K. denitrificans is emended to indicate that maltose is not utilized.

Antimicrobial susceptibility testing of Francisella tularensis with a modified Mueller-Hinton broth.

A modified Mueller-Hinton broth was developed to perform antimicrobial susceptibility tests on Francisella tularensis. Adequate growth of the organism was obtained within 24 h of inoculation, and MICs could be read at that time. We tested 15 selected strains of F. tularensis and five reference quality control strains in this medium with 36 antimicrobial agents. The MICs of the aminoglycosides and tetracycline increased 1 to 3 dilutions in this medium compared with those in the usual medium, but the other antimicrobial agents were not consistently affected by the medium. Even though the medium caused an increase in MICs, the aminoglycosides and tetracyclines remained very active in vitro against F. tularensis. Other antimicrobial agents effective in vitro were chloramphenicol, erythromycin, ceftazidime, moxalactam, cefotaxime, ceftriaxone, and Sch 29482 (a cephalosporin).

Characterization and differentiation of 59 strains of Moraxella urethralis from clinical specimens.

The biochemical characteristics of 59 strains of Moraxella urethralis from clinical specimens, primarily from urine and the female genital tract, were studied. The characteristics included (i) the inability to acidify carbohydrate substrates, (ii) the ability to produce phenylalanine deaminase, (iii) the ability to reduce nitrite, (iv) the lack of urease activity, and (v) the ability of most strains to alkalinize citrate. A means of differentiating M. urethralis from Moraxella osloensis and Moraxella phenylpyruvica was determined.

Cellular fatty acid composition of group IVe, a nonsaccharolytic organism from clinical sources.

The cellular fatty acid composition of a group of gram-negative nonfermentative organisms designated group IVe was studied by gas-liquid chromatography. Strains of this group are isolated most frequently from urine and most closely resemble the Alcaligenes in conventional biochemical tests. On the basis of cellular fatty acids, however, we found these organisms to be strikingly different from Alcaligenes and other gram-negative species with similar phenotypic characteristics. The gas-liquid chromatography procedure offers an additional diagnostic test for rapid identification of unclassified bacteria like group IVe.

Chronic granulomatous disease: fatal septicemia caused by an unnamed gram-negative bacterium.

A 2-year-old boy with proven X-linked chronic granulomatous disease was placed under continuous co-trimoxazole prophylaxis. He remained free of infection for 4 years. At age 6.25 years, he suddenly developed a fever with no localizing signs and died 16 days later in septic shock. A gram-negative, catalase-positive, halophilic, aerobic bacterium was cultured from blood, bone marrow, and ascitic fluid. This organism could not be identified in microbiological laboratories in Europe and the United States. Its biochemical features indicate that it may belong to a species which has not previously been described.

Phenotypic characteristics, fatty acid composition, and isoprenoid quinone content of CDC group IIg bacteria.

Eleven strains of eugonic, nonoxidative, gram-negative rods isolated from clinical specimens formed a distinct group that was designated CDC group IIg. Five of the 11 isolates were from wounds. The phenotypic characteristics of CDC group IIg were most similar to those of Weeksella species, with the major difference being that CDC group IIg strains grew on MacConkey agar in 1 to 2 days, did not hydrolyze gelatin, and did not produce urease. All 11 strains of CDC group IIg possessed a distinct fatty acid profile that was characterized by large amounts (19 to 29%) of 18:1 omega 7c, 16:0, and 16:1 omega 7c, moderate amounts (6 to 10%) of 3-OH-14:0 and 14:0, and smaller amounts (1 to 2%) of 18:2, 18:0, and 3-OH-16:0. This fatty acid profile differs from those of Weeksella species by the absence of branched-chain fatty acids. CDC group IIg contains ubiquinone-8, as opposed to menaquinone-6 in Weeksella species. The isolates were susceptible to a variety of antimicrobial agents, including the aminoglycosides, tetracyclines, quinolones, sulfonamides, and polymyxin B.

Exudative pharyngitis possibly due to Corynebacterium pseudodiphtheriticum, a new challenge in the differential diagnosis of diphtheria.

Corynebacterium pseudodiphtheriticum has rarely been reported to cause disease in humans, despite its common presence in the flora of the upper respiratory tract. We report here a case of exudative pharyngitis with pseudomembrane possibly caused by C. pseudodiphtheriticum in a 4-year-old girl. The case initially triggered clinical and laboratory suspicion of diphtheria. Because C. pseudodiphtheriticum can be easily confused with Corynebacterium diphtheriae in Gram stain, clarification of its role in the pathogenesis of exudative pharyngitis in otherwise healthy persons is of public health importance. Simple and rapid screening tests to differentiate C. pseudodiphtheriticum from C. diphtheriae should be performed to prevent unnecessary concern in the community and unnecessary outbreak control measures.

Use of the rapid fermentation test in determining carbohydrate reactions of fastidious bacteria in clinical laboratories.

The rapid fermentation test was used to determine the carbohydrate reactions of some of the fastidious bacteria encountered in clinical laboratories, such as: Haemophilus species, including Haemophilus vaginalis; Actinobacillus actinomycetemcomitans; Cardiobacterium hominis; Kingella species; Corynebacterium species; Propionibacterium species; and Erysipelothrix rhusiopathiae. Results were usually obtained within 4 h by using inocula from 24- or 48-h blood or chocolate agar media.

Cellular fatty acids of Brucella canis and Brucella suis.

The cellular fatty acid composition of Brucella canis and Brucella suis was determined by gas-liquid chromatography. The presence of relatively large amounts of a 19-carbon cyclopropane fatty acid in B. suis was a major distinguishing feature between these organisms. The gas-liquid chromatography test for cellular fatty acids provides an additional criterion for the distinction of antigenically rough strains of B. suis which cannot be differentiated from B. canis by conventional procedures.

Chemical characterization of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf.

The cellular fatty acid, sphingolipid, and isoprenoid quinone compositions of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf were determined, using thin-layer, gas-liquid, and reverse-phase high-performance liquid chromatography. The fatty acid data showed that groups IIe, IIh, and IIf were similar to recognized Flavobacterium species by the presence of relatively large amounts of iso-branched hydroxy and nonhydroxy acids. Groups IIe and IIh were essentially identical in fatty acid composition but were distinguished from group IIf, F. breve, and F. odoratum on the basis of minor qualitative and quantitative differences. All strains tested contained menaquinone 6 as the major isoprenoid quinone, and all lacked sphingolipids. Overall, the chemical data suggest that groups IIe, IIh, and IIf are additional Flavobacterium species and are different from sphingobacteria, which contain sphingolipid and menaquinone 7 as the major quinone.