In vivo muscle 31P nuclear magnetic resonance spectroscopy during treatment of halothane-sensitive and halothane-nonsensitive pigs.

In vivo muscle 31P nuclear magnetic resonance spectroscopy was performed on 10 female pigs originating from a homozygous halothane-sensitive line and on 10 female pigs from a homozygous halothane-nonsensitive line. The mean concentration of phosphocreatine in the biceps femoris muscle of the anesthetized pigs decreased to 86% of the initial value after 11 minutes of halothane exposure (3%, oxygen flow 3 L/min). After the next 5.6 minutes, phosphocreatine concentration reached a minimal value of 52%, followed by a mean recovery to 76% of the initial value during the ensuing 11 minutes. Response was not observed in anesthetized homozygous halothane-nonsensitive pigs. Thus, a decrease to 86% of the initial value of phosphocreatine was 100% predictive for homozygous halothane-sensitive pigs with body weight ranging from 10 to 18 kg.

Transfer of several phytopathogenic Pseudomonas species to Acidovorax as Acidovorax avenae subsp. avenae subsp. nov., comb. nov., Acidovorax avenae subsp. citrulli, Acidovorax avenae subsp. cattleyae, and Acidovorax konjaci.

DNA-rRNA hybridizations, DNA-DNA hybridizations, polyacrylamide gel electrophoresis of whole-cell proteins, and a numerical analysis of carbon assimilation tests were carried out to determine the relationships among the phylogenetically misnamed phytopathogenic taxa Pseudomonas avenae, Pseudomonas rubrilineans, "Pseudomonas setariae," Pseudomonas cattleyae, Pseudomonas pseudoalcaligenes subsp. citrulli, and Pseudomonas pseudoalcaligenes subsp. konjaci. These organisms are all members of the family Comamonadaceae, within which they constitute a separate rRNA branch. Only P. pseudoalcaligenes subsp. konjaci is situated on the lower part of this rRNA branch; all of the other taxa cluster very closely around the type strain of P. avenae. When they are compared phenotypically, all of the members of this rRNA branch can be differentiated from each other, and they are, as a group, most closely related to the genus Acidovorax. DNA-DNA hybridization experiments showed that these organisms constitute two genotypic groups. We propose that the generically misnamed phytopathogenic Pseudomonas species should be transferred to the genus Acidovorax as Acidovorax avenae and Acidovorax konjaci. Within Acidovorax avenae we distinguished the following three subspecies: Acidovorax avenae subsp. avenae, Acidovorax avenae subsp. cattleyae, and Acidovorax avenae subsp. citrulli. Emended descriptions of the new taxa are presented.

Revision of Campylobacter, Helicobacter, and Wolinella taxonomy: emendation of generic descriptions and proposal of Arcobacter gen. nov.

Hybridization experiments were carried out between DNAs from more than 70 strains of Campylobacter spp. and related taxa and either 3H-labeled 23S rRNAs from reference strains belonging to Campylobacter fetus, Campylobacter concisus, Campylobacter sputorum, Campylobacter coli, and Campylobacter nitrofigilis, an unnamed Campylobacter sp. strain, and a Wolinella succinogenes strain or 3H- or 14C-labeled 23S rRNAs from 13 gram-negative reference strains. An immunotyping analysis of 130 antigens versus 34 antisera of campylobacters and related taxa was also performed. We found that all of the named campylobacters and related taxa belong to the same phylogenetic group, which we name rRNA superfamily VI and which is far removed from the gram-negative bacteria allocated to the five rRNA superfamilies sensu De Ley. There is a high degree of heterogeneity within this rRNA superfamily. Organisms belonging to rRNA superfamily VI should be reclassified in several genera. We propose that the emended genus Campylobacter should be limited to Campylobacter fetus, Campylobacter hyointestinalis, Campylobacter concisus, Campylobacter mucosalis, Campylobacter sputorum, Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and "Campylobacter upsaliensis." Wolinella curva and Wolinella recta are transferred to the genus Campylobacter as Campylobacter curvus comb. nov. and Campylobacter rectus comb. nov., respectively. Bacteroides gracilis and Bacteroides ureolyticus are generically misnamed and are closely related to the genus Campylobacter. Campylobacter nitrofigilis, Campylobacter cryaerophila, and an unnamed Campylobacter sp. strain constitute a new genus, for which the name Arcobacter is proposed; this genus contains two species, Arcobacter nitrofigilis comb. nov. (type species) and Arcobacter cryaerophilus comb. nov. Wolinella succinogenes so far is the only species of the genus Wolinella. The genus Helicobacter is also emended; Campylobacter cinaedi and Campylobacter fennelliae are included in this genus as Helicobacter cinaedi comb. nov. and Helicobacter fennelliae comb. nov., respectively. The genus "Flexispira," with "Flexispira rappini" as the only species, is closely related to the genus Helicobacter. The free-living, sulfur-reducing campylobacters do not belong to any of these genera; they probably constitute a distinct genus within rRNA superfamily VI.

Acidovorax, a new genus for Pseudomonas facilis, Pseudomonas delafieldii, E. Falsen (EF) group 13, EF group 16, and several clinical isolates, with the species Acidovorax facilis comb. nov., Acidovorax delafieldii comb. nov., and Acidovorax temperans sp. nov.

Pseudomonas facilis and Pseudomonas delafieldii are inappropriately assigned to the genus Pseudomonas. They belong to the acidovorans rRNA complex in rRNA superfamily III (i.e., the beta subclass of the Proteobacteria). The taxonomic relationships of both of these species, two groups of clinical isolates (E. Falsen [EF] group 13 and EF group 16), and several unidentified or presently misnamed strains were examined by using DNA:rRNA hybridization, numerical analyses of biochemical and auxanographic features and of fatty acid patterns, polyacrylamide gel electrophoresis of cellular proteins, and DNA:DNA hybridization. These organisms form a separate group within the acidovorans rRNA complex, and we propose to transfer them to a new genus, Acidovorax. We describe the following three species in this genus: the type species, Acidovorax facilis (formerly Pseudomonas facilis), with type strain LMG 2193 (= CCUG 2113 = ATCC 11228); Acidovorax delafieldii (for the former Pseudomonas delafieldii and most of the EF group 13 strains), with type strain LMG 5943 (= CCUG 1779 = ATCC 17505); and Acidovorax temperans (for several former Pseudomonas and Alcaligenes strains and most of the EF group 16 strains), with type strain CCUG 11779 (= LMG 7169).

Identification of Campylobacter cinaedi isolated from blood and feces of children and adult females.

Five Campylobacter-like organisms isolated from blood and feces were identified by numerical analysis of gel electrophoretic protein profiles and immunotyping as Campylobacter cinaedi. Two of these strains were isolated from adult females; the remaining three strains were isolated from children, two of whom were girls. C. cinaedi has hitherto been isolated only from rectal swabs and blood of homosexual and bisexual males with gastrointestinal symptoms. The results presented extend our knowledge of the features and the habitat of C. cinaedi.

Inter- and intrafamilial similarities of rRNA cistrons of the Pasteurellaceae.

We performed hybridizations between labeled rRNAs from seven representative members of the family Pasteurellaceae and from three other taxa on the one hand and DNAs from 53 strains known or presumed to belong to the Pasteurellaceae on the other hand. The members of the Pasteurellaceae are most closely related to members of the Enterobacteriaceae, the Vibrionaceae, the Aeromonadaceae, and the genus Alteromonas. The family Pasteurellaceae is very heterogeneous. There are at least seven rRNA branches. Several organisms with the same genus name are dispersed over the entire dendrogram. The "Histophilus ovis," [Haemophilus] ducreyi, [Actinobacillus] actinomycetemcomitans, and [Haemophilus] aphrophilus rRNA branches are separate and quite remote from the three authentic genera in this family; this might justify eventual later separate generic status. DNA-rRNA hybridization with suitable, labeled rRNA probes is an excellent method to establish whether an organism belongs in the Pasteurellaceae; e.g., some strains of Bisgaard's taxa 7, 13, and 16 and of the gas-producing "SP" group certainly belong in this family, whereas three bovine lymphangitis organisms (strains NCTC 10547, NCTC 10549, and NCTC 10553), [Haemophilus] piscium ATCC 10801T (T = type strain), and [Pasteurella] piscicida ATCC 17911 belong in the Enterobacteriaceae, the Aeromonadaceae, and the Vibrionaceae, respectively.

Skeletal muscularity and heart function in growing piglets.

Nine female piglets (about 15 kg), originating from a line with a low skeletal muscularity, were compared to 9 female piglets (about 15 kg) originating from a line with a high skeletal muscularity. Electrocardiogram-parameters (frequency rise, ST-elevation, heart rate expressed as mean values per h) were measured for 17 h with an ambulatory Holter-device. Dimensions of longissimus dorsi muscle (depth, circumference, surface, speed of ultrasound) were measured with an ultrasound device based on the pulse echo principle. Parameters, with respect to muscular dimensions, were statistically significant, differing between both lines, whereas, only the ST-elevation was significant, lower in the high skeletal muscularity line. The lower muscular depth was, the higher the ST-elevation and the higher the mean lowest heart rate were, and the lower the speed of ultrasound through the muscle tissue, by the higher the mean frequency rise. Hence, in growing piglets of about 15 kg, an impairment of heart function coincides gradually with a proportional higher development of skeletal muscularity.

Identification of EF group 22 campylobacters from gastroenteritis cases as Campylobacter concisus.

EF (E. Falsen) group 22, a group of Campylobacter strains sorted out by routine immunotyping among unidentified or misidentified human nonoral clinical specimens, was characterized by numerical analysis of gel electrophoretic protein profiles and immunotyping. The protein electrophoretic and immunotyping analyses, DNA:DNA hybridizations, and the DNA base composition demonstrated unambiguously that all EF group 22 strains belong to Campylobacter concisus. EF group 22 strains have DNA binding values of at least 42% with the type strain of C. concisus, showing a considerable degree of genomic heterogeneity. The isolation from blood, esophagus, stomach, duodenum, and feces of humans in association with different gastrointestinal disorders considerably extends the clinical significance of this species. Our results indicate that sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunotyping are excellent tools for the identification of the fastidious C. concisus strains and relatives.

Specific Neisseria gonorrhoeae DNA-probes derived from ribosomal RNA.

Eighteen sequences complementary to less-conserved regions within the 16S and 23S ribosomal ribonucleic acid (rRNA) of Neisseria gonorrhoeae were subcloned or chemically synthesized and used as probes in a dot-spot deoxyribonucleic acid (DNA): DNA hybridization format. Some of these probes exclusively detected Neisseria gonorrhoeae nucleic acid, whereas others also showed hybridization signals with nucleic acid from other bacterial species. Our results indicate that rRNA-derived DNA-probes can be used to differentiate between very closely related taxa without the use of Southern-blot analysis.

Separation of Escherichia adecarboxylata from the "Erwinia herbicola-Enterobacter agglomerans" complex and from the other Enterobacteriaceae by nucleic acid and protein electrophoretic techniques.

The species Escherichia adecarboxylata was examined for DNA relatedness to the "Erwinia herbicola-Enterobacter agglomerans" complex and to other members of the family Enterobacteriaceae. DNA-DNA hybridizations (nitrocellulose filter method) showed that strains received as E. adecarboxylata were highly related to each other (73-100% homology). Three strains of E. agglomerans and one strain of E. herbicola showed, respectively, 77, 96, 97 and 92% relatedness with the labelled DNA of E. adecarboxylata. Two groups (E2 and E3) of "atypical coliforms" previously described by Gavini et al. (1983) showed high reassociation values (76-79% and 80-89%, respectively) with E. adecarboxylata. Most of these strains produced similar or nearly identical protein electrophoregrams. All these strains were therefore classified in E. adecarboxylata. This taxon yielded hybridization values lower than 53% with the previously described phenetic or genetic groups belonging to or related to the "herbicola-agglomerans" complex and values lower than 64% with 56 other species of the Enterobacteriaceae. It was concluded that E. adecarboxylata is a species different from E. agglomerans and the other species of the family Enterobacteriaceae. A new definition of the species E. adecarboxylata is presented.

Differentiation between Xanthomonas campestris pv. oryzae, Xanthomonas campestris pv. oryzicola and the bacterial 'brown blotch' pathogen on rice by numerical analysis of phenotypic features and protein gel electrophoregrams.

Thirty-five Xanthomonas campestris pv. oryzae, fourteen X. campestris pv. oryzicola strains and six 'brown blotch' pathogens of rice, all of different geographical origin, were studied by numerical analysis of 133 phenotype features and gel electrophoregrams of soluble proteins, %G + C determinations and DNA:rRNA hybridizations. The following conclusions were drawn. (i) The Xanthomonas campestris pathovars oryzae and oryzicola display clearly distinct protein patterns on polyacrylamide gels and can be differentiated from each other by four phenotype tests. (ii) Both pathovars are indeed members of Xanthomonas which belongs to a separate rRNA branch of the second rRNA superfamily together with the rRNA branches of Pseudomonas fluorescens, Marinomonas, Azotobacter, Azomonas and Frateuria. (iii) 'Brown blotch' strains are considerably different from X. campestris pv. oryzae and oryzicola. They are not members of the genus Xanthomonas, but are more related to the generically misnamed. Flavobacterium capsulatum, Pseudomonas paucimobilis, Flavobacterium devorans and 'Pseudomonas azotocolligans' belonging in the fourth rRNA superfamily. (iv) No correlation was found between the virulence, pathogenic groups or geographical distribution of X. campestris pv. oryzae or oryzicola strains and any phenotypic or protein electrophoretic property or clustering.

Identification of Acetobacter strains isolated from spoiled lactic acid fermented meat food for pets.

Five Acetobacter isolates from lactic acid fermented meat food for pets were characterized by 177 morphological, physiological and biochemical traits. Four isolates were identified as A. pasteurianus, one as A. aceti. It is emphasized that access of such bacteria to lactic acid fermented foods should be avoided.

The catabolism of 3-ketolactose in Agrobacterium.

The existence of a novel enzyme, catalyzing the hydrolysis of 3-ketolactose, is described in the different genetic groups of the genus Agrobacterium. The enzyme differs from the other glycosidases formed by Agrobacterium during growth on lactose. The inducibility of the enzyme could be demonstrated in a strain from biotype 1, but not in a strain from biotype 2. The specific activity of the 3-ketolactose hydrolase is higher in extracts of strains belonging to biotype 2 than to biotype 1. The optimum pH of the enzyme in Tris-HCl buffer is 8.4-8.5. The K(m) value for 3-ketolactose is 2.2 × 10(-2) M. The 3-ketolactose hydrolase is stimulated by Mg(2+) and Mn(2+), and inhibited by Zn(2+). Mercaptoethanol promotes the reaction rate in extracts of strains of biotype 1 but not in extracts of strains of bio-type 2.

Numerical Analysis of Phenotypic Features and Protein Gel Electrophoregrams of a Wide Variety of Acetobacter strains. Proposal for the Improvement of the Taxonomy of the Genus Acetobacter Beijerinck 1898, 215.

Ninety-eight strains, representing all Acetobacter species and subspecies from the Approved Lists of Bacterial Names (Skerman et al., 1980), were examined in a numerical analysis of 177 phenotypic features and compared to ninety-eight Gluconobacter and seven Frateuria strains. Four phenons could be delineated, corresponding to Frateuria (phenon 1), A. aceti subsp. liquefaciens (phenon 2), Gluconobacter (phenon 3) and Acetobacter minus A. aceti subsp. liquefaciens (phenon 4). Acetobacter, Frateuria and Gluconobacter are well- could be distinguished. Comparison of the protein electrophoregrams of Acetobacter strains revealed a fairly high internal homogeneity within phenon 2, subphenons C and D. Strains of the subphenon E gave very divergent protein patterns. The following classificatory changes are proposed within the genus Acetobacter: (1) Acetobacter liquefaciens sp. nov. is proposed for the homogeneous phenon 2, containing all 12 A. aceti subsp. liquefaciens strains (% G + C range of 62.3 to 64.6; IAM 1834 as type strain); (2) for the homogeneous subphenon D containing 8 A. aceti subsp. aceti strains, the name Acetobacter aceti emend, should be retained (% G + C range of 55.9 to 59.5; NCIB 8621 as type strain); (3) for subphenon E, a heterogeneous group, containing a variety of Acetobacter subspecies (all with their type strain) the species name Acetobacter pasteurianus emend, is preserved with LMD 22.1 as type strain; this species has the broad % G + C range of 52.8 to 62.5; (4) for subphenon C, a new species, Acetobacter hansenii sp. nov. is proposed (% G + C range of 58.1 to 62.6, NCIB 8746 as type strain). Minimal descriptions and differentiating keys are provided.

The nitrogen requirements of Gluconobacter, Acetobacter and Frateuria.

The nitrogen requirements of 96 Gluconobacter, 55 Acetobacter and 7 Frateuria strains were examined. Only some Frateuria strains were able to grow on 0.5% yeast extract broth or 0.5% peptone broth. In the presence of D-glucose or D-mannitol as a carbon source, ammonium was used as the sole source of nitrogen by all three genera. With ethanol, only a few Acetobacter strains grew on ammonium as a sole nitrogen source. Single L-amino acids cannot serve as a sole source of carbon and nitrogen for growth of Gluconobacter, Acetobacter or Frateuria. The single L-amino acids which were used by most strains as a sole nitrogen source for growth are: asparagine, aspartic acid, glutamine, glutamic acid, proline and alanine. Some Acetobacter and Gluconobacter strains deaminated alanine, asparagine, glutamic acid, threonine, serine and proline. No Frateuria strain was able to develop on cysteine, glycine, threonine or tryptophan as a sole source of nitrogen for growth. An inhibitory effect of valine may explain the absence of growth on this amino acid. No amino acid is "essential" for Gluconobacter, Acetobacter or Frateuria.

Gluconobacters from honey bees.

Fifty-six Gluconobacter strains and one Acetobacter strain were isolated from honey bees and their environment in three different regions in Belgium and identified phenotypically. Polyacrylamide gel electrophoresis of the soluble cell proteins showed that two different types exist within the Gluconobacter isolates: strains from type A were found in samples of the three regions, whereas strains from type B were only isolated in two of the three regions. Both types could occur in bees from the same region, from several hives of one bee keeper and from one hive. Strains from type A were almost identical with collection strain G. oxydans subsp. suboxydans NCIB 9018, whereas strains from type B constituted a new protein electrophoretic type within the genus Gluconobacter. Although Gluconobacter is apparently associated with honey bees, it is not known whether it is important or required for the bees or any hive product.

Cytochromes c-556 from three genetic races of Agrobacterium. Purification and comparison of their properties.

1. The soluble cytochromes c-556 from three strains of Agrobacterium tumefaciens, B6, II Chrys and Apple 185 have been purified to homogeneity. The strains are representative members of the three main genetic races of Agrobacterium. The purity of the final preparations was established by electrophoresis with an without sodium dodecyl sulphate, by analytical isoelectric focusing and ultracentrifugation, and by N-terminal analysis. 2. Properties of these cytochromes were compared wih those of cytochrome c-556 from A. tumefaciens, strain B2a, a member of the same genetic race as strain B6. The four cytochromes are monohaem proteins with molecular weights of about 12300 (determined by four different methods). The isoelectric points of those from strains B6 and B2a are identical at pH 5.5, but they differ from the cytochromes of the other genetic races: cytochrome c-556 from strain Apple 185 is more acidic (ph 5.2) and that from strain II Chrys more basic (pH 6.2). The cytochromes from strains b6 and B2a have very similar but not identical amino acid compositions; both of them differ more from Apple 185 than from II Chrys c-556. 3. Comparison of the tryptic, chymotryptic and thermolytic fingerprints of cytochrome c-556 from strains B2a and II Chrys reveals strong homology between the primary structures of these cytochromes. Therefore and because of the sequence identity of the first eight residues, the cytochromes c-556 from strains II Chrys, B6 and B2a are most likely C-terminal haem-bound, of the same type as the cytochrome c' from photosynthetic bacteria.