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Zusammenfassung Bei der mikrobiologischen Analyse eines Ausbruchs respiratorischer Erkrankungen bei albinotischen Sprague-Dawley-Ratten, die konventionell gehalten wurden, wurde aus pneumonischem Gewebe eine neue Pasteurella species isoliert. Dieser Keim ahnelt phanotypisch Pasteurella gallinarum, gehort aber einer anderen DNS-Homologie-Gruppe an. Kulturen der neuen Species wurden unter den Nummern 11 051 und 11 052 in der National Collection of Type Cultures, London hinterlegt. Die neue Pasteurella sp. erwies sich bei SPF-Ratten nach intranasaler wie nach parenteraler Zufuhrung als hochgradig pneumotrop; sie war jedoch nicht imstande, Erkrankungen auszulosen. Die untersuchte Rattenpopulation war ferner Trager von Pasteurella pneumotropica, ebenso wie zwei andere konventionell gezuchtete Rattenstamme, die zu Vergleichszwecken untersucht wurden. Uber die phanotypische Variabilitat der untersuchten P. pneumotropica-Stamme wird berichtet. Summary Mixed infection of albino rats by Pasteurella pneumotropica and a new pneumotropic Pasteurella species In the microbiological analysis of an outbreak of respiratory disease in albino Sprague-Dawley rats kept under conventional conditions, a new Pasteurella species was isolated from pneumonic tissue. This organism was phenotypically related to Pasteurella gallinarum but was shown to belong to a different DNA homology group. Cultures of the new species have been deposited in the National Collection of Type Cultures, London under numbers 11051 and 11052 respectively. The new Pasteurella sp. was highly pneumotropic in SPF rats after intranasal or parenteral application; it was however not able to induce disease. Furthermore, the rat population investigated carried Pasteurella pneumotropica, as did two other conventional rat strains investigated for comparison. The phenotypic variability of the P. pneumotropica strains studied is reported. Resume Infection mixte de rats albinos par Pasteurella pneumotropica et une nouvelle Pasteurella species pneumotrope Une nouvelle Pasteurella species a ete isolee a partir des tissus pulmonaires lors d'analyses microbiologiques chez des rats Sprague-Dawley gardes conventionnellement et atteints d'affections respiratoires. Ce germe ressemble au phenotype Pasteurella gallinarum mais appartient a un autre groupe ADN homologue. Des cultures ont ete deposees a la National Collection of Type Cultures de Londres sous les numeros 11051 et 11052. La nouvelle Pasteurella sp. s'est revelee hautement pneumotrope lors d'injections intranasales et parenterales chez des rats SPF; elle n'a pas ete capable cependant de declencher des symptomes de maladie. La population de rats examinee etait egalement porteuse de Pasteurella pneumotropica ainsi que deux autres souches de rats elevees conventionellement et examinees comme temoins. La variabilite phenotypique des souches de P. pneumotropica examinees est decrite. Resumen Infeccion mixta en ratas albinas por Pasteurella pneumotropica y una especie Pasteurella pneumotropa nueva En el analisis microbiologico de un brote de enfermedades respiratorias en ratas albinoides Sprague-Dawley, recriadas de forma convencional, se aislo de tejido pneumonico una especie Pasteurella nueva. Este germen se asemeja fenotipicamente a Pasteurella gallinarum, aunque pertenece a otro grupo de homologia ADN. En la National Collection of Type Cultures, Londres, han sido consignados cultivos de la especie nueva. La Pasteurella sp. nueva se mostro como pneumotropa extrema en ratas libres de germenes patogenos especificos tras administracion intranasal o parenteral; sin embargo, no fue capaz de desencadenar enfermedades. La poblacion de ratas examinada tambien era portadora de Pasteurella pneumotropica, a la par que otras dos estirpes de ratas criadas al modo convencional, las cuales se examinaron con fines comparativos. Se informa sobre la variabilidad fenotipica de las estirpes P. pneumotropica estudiadas.

Evidence was obtained to indicate that equine strains of organisms previously described as Actinobacillus suis or hemolytic variants of Actinobacillus equuli might constitute a separate group of organisms provisionally designated taxon 11. Four biovars were noticed within taxon 11. Selected DNA:DNA hybridizations support the classification of the mannitol positive biovar 2 of taxon 11 distinct from porcine A. suis. The final taxonomical position of taxon 11, however, has to await more detailed genetic studies including all biovars of taxon 11. A species name has not been suggested for the same reasons. The present observations also indicate that strains identified as taxon 11 apparently constitute a part of the normal bacterial flora in the oral cavity of horses.

Ten catalase-positive isolates and one catalase-negative isolate that had been assigned to Eikenella corrodens were compared to the nomenclatural type strain regarding selected phenotypic and molecular features and chromosomal deoxyribonucleic acid (DNA) relatedness using the spectrophotometric method. Five catalase-positive human isolates were assigned to the genomic species Eikenella corrodens on the basis of high DNA relatedness levels. Three others, among them strain Chen UB 204, exhibited only moderate degrees of DNA relatedness to the type strain and with each other. Two catalase-positive isolates from dogs were closely interrelated, but yielded only low degrees of DNA binding with Eikenella corrodens and the Eikenella-like human isolates. These findings confirm that the human eikenellas comprise more than one genomic species and that the canine strains represent a distinct taxonomic entity. The differentiation of the strains investigated by conventional phenotypic features, hydrolytic enzyme reactions, and cellular carbohydrate patterns was considered.

Summary Selected strains representing established and newly described taxa in the family Pasteurellaceae were investigated for their cellular lipid and carbohydrate composition to clarify the taxonomic significance of such features. Methylated cellular fatty acids and acetylated derivatives of the cellular carbohydrates were determined by capillary gas chromatography using a flame ionization detector. In part the carbohydrates were identified by mass spectrometry. Phospholipids were determined by thin layer chromatography, the lipoquinones by high pressure liquid chromatography. The cellular fatty acid patterns proved to be uniform with minor variations, but the separation from the Neisseriaceae and from Moraxella was possible. Also the distribution of the phospholipids was uniform within the family. The lipoquinone contents were useful for the discrimination of groups within the family not necessarily reflecting the degree of genomic relatedness. The analysis of the cellular carbohydrates resulted in a common sugar pattern with all members of the family and characteristic carbohydrate profiles discriminating groups, often to the species level. All of the cytochemical features considered were useful for the characterization of the family Pasteurellaceae .

Pasteurella species and related taxa are opportunistic pathogens parasitizing on mucous membranes of higher organisms containing sialic acids. Therefore, sialidase is a virulence factor which up to now has been described to be present in P. haemolytica, P. multocida, and P. volantium. Because of some taxonomic changes and the description of many new species or still unnamed groups, the presence of sialidase and the metabolic successor enzyme, N-acetylneuraminate lyase, was investigated in 65 Pasteurella or Pasteurella-like strains. The detection of enzymes was performed by colorimetry, by paper chromatography and immunoelectrophoresis. Using bovine submaxillary mucin as substrate, sialidases were produced in all strains studied although the activities were different. Most strains but not all were positive in N-acetylneuraminate lyase, too. Taken together, the strains of Pasteurella sensu stricto showed the strongest activities of sialidase, those of the Pasteurella aerogenes complex the lowest. However, because of loss of sialidase activity during subcultivation, there is little feasibility to characterize Pasteurella species by these enzymes.

Summary Twelve human isolates exhibiting the properties of the family Pasteurellaceae but phenotypically deviating from established species, or resembling species of animal origin that are only rarely reported to occur in human materials, were checked for their identities by DNA-DNA hybridization. The collection consisted of one strain of Actinobacillus lignieresii , two strains of Actinobacillus hominis (mannose-positive), two hitherto undescribed Actinobacillus or Actinobacillus -like species, [ Pasteurella ] haemolytica biovar T, CDC group HB-5 (initially oxidase and indole-negative), a new species in the [ Haemophilus ] aphrophilus /[ Haemophilus ] segnis group, a new sucrose-negative and gas-producing Pasteurella -like species, and three strains of Bisgaard's Pasteurella -like taxon 16. Some diagnostically useful features of these unusual human Pasteurellaceae are described.

The carbohydrate patterns, isoprenoid quinones, fatty acids and phospholipids of the species of the genus Pasteurella sensu stricto were investigated to evaluate their taxonomic significance and their applicability for the identification of these bacteria. Forty-six representative strains of the 11 species of Pasteurella were examined. The data obtained indicated that the carbohydrate patters are species or subspecies specific and may, therefore, become an important and useful diagnostic tool. Fatty acids and phospholipids showed a feature characteristic of the members of the genus and the isoprenoid quinones exhibited a mostly genus-specific feature with remarkable quantitative differences.

Clostridium La 1 obtained from a Clostridium kluyveri culture was compared with a typical C. kluyvery strain (DSM 555). The former grows on cortonate and is unable to use ethanol-acetate as carbon sources. The latter grows on crotonate only after long adaptation periods. Resting cells of both strains show also pronounced differences in the fermentation of crotonate. This holds even for C. kluyveri grown on crotonate. Besides several other differences the most striking is that there is no hybridization between the DNA of both strains. Crotonate seems not to be a very special carbon source since C. butyricum and C. pasteurianum grow on crotonate medium supplemented by peptone and yeast extract.

The phylogenetic relationships among strains of Pasteurella multocida representing Carter's serogroups A, B, (C), D and E, the type strain (which also represents serogroup A, and biovar 4), the indole-negative strain Schutze HS, and two “dog-type” strains (biovar 6) were investigated by DNA:DNA hybridization using the optical method. The genome DNAs of the “dog-type” strains were almost identical; they displayed, however, only 20 % binding with the DNA of the type strain of P. multocida , and even lower or no measurable binding with the DNAs of the other strains tested. The taxon hitherto classified as biovar 6 of P. multocida is therefore considered as a distinct species; yet the data so far available do not rule out that it belongs to a genus other than Pasteurella sensu stricto . — The remaining strains exhibited high genome DNA relatedness, with between 64 and 98% DNA binding. The present data do not rule out the existence of molecular subspecies in P. multocida .

Ten epidemiologically independent Capnocytophaga strains from clinical materials constituted three distinct DNA homology groups corresponding to the established species, Capnocytophaga ochracea, sputigena, and gingivalis. The three groups exhibited only low, or even insignificant degrees of genomic relatedness mutually, and to a reference culture of CDC group DF-2. All of five Capnocytophaga ochracea strains considered were involved in topic infections whereas four strains, isolated from blood belonged to either Capnocytophaga sputigena or Capnocytophaga gingivalis. However, Capnocytophaga ochracea and Capnocytophaga sputigena could not be discriminated on the basis of DNA base composition, genome size, or phenotypic features including cellular fatty acids, lipoquinones, and various fermentation and hydrolase reactions. The need for additional phenotypic criteria characterizing species of Capnocytophaga is underlined by the present findings.

SUMMARY Five species and two subspecies of nonmotile and coccoid Achromobacteres are described, including the organisms hitherto known as Bacterium anitratum Schaub and Hauber and Alcaligenes metalcaligenes Castellani and Chalmers. They are Achromobacter mucosus (von Lingelsheim 1906) Mannheim and Stenzel 1962, A. conjunctivae Mannheim and Stenzel 1962, A. haemolyticus Mannheim and Stenzel 1962, A. haemolyticus subsp. alcaligenes Mannheim and Stenzel 1962, A. metalcaligenes (Castellani and Chalmers 1919) Bergey et al. 1925, and A. citroalcaligenes Mannheim and Stenzel 1962.

Summary The type strain of Eikenella corrodens ( Eiken 1958) Jackson and Goodman 1972 and eleven epidemiologically independent clinical isolates recovered from periodontal locations, putrid wounds, abscesses, and bacteraemias were investigated for their genomic relationships by DNA-DNA hybridization with the renaturation method, genome molecular complexity, DNA base composition and some phenotypic features. The bacterial strains studied were interrelated at or above the 80% DNA binding level, their chromosomal DNAs exhibiting a mean molecular mass of 1.7 × 10 9 daltons and a mean guanine plus cytosine content of 55.1 mol%. Variations in colonial morphology, hemolytic activity on sheep blood agar, reduction of nitrates, oxidation of carbohydrates, lipase, leucine, valine, and cystine aminopeptidase and acid phosphatase activities occurred among closely interrelated strains. The definition of the species and current identification keys must be emended accordingly.

Summary: The respiratory quinones of 73 strains of Gram-positive bacteria including spore-forming rods, lactic-acid bacteria and actinomycetes were examined. Menaquinones with seven isoprenoid units (MK-7) were the main quinone type found in representatives of the genus Bacillus and in Sporolactobacillus inulinus. However, a strain of B. thuringiensis produced MK-8 in addition to MK-7, and strains of B. lentus and B. pantothenticus appeared to produce MK-9 and MK-8, respectively, with no MK-7. In the Clostridia and lactic-acid bacteria, no quinones were found, except in Pediococcus cerevisiae NCTC 8066 and Lactobacillus casei subsp. rhamnosus ATCC 7469, which contained menaquinones, and Streptococcus faecalis NCTC 775 and HIM 478-1, which contained demethylmenaquinones, in relatively low concentrations. Menaquinones were also found in the actinomycetes (except Actinomyces odontolyticus and Bifidobacterium bifidum which did not produce any quinones) and in Protaminobacter alboflavus ATCC 8458, the so-called Actinobacillus actinoides ATCC 15900 and Noguchia granulosis NCTC 10559.

The taxonomy of five typical human isolates of Pasteurella ureae , one strain of Actino-bacillus hominis , and three murine isolates which had been designated as Pasteurella ureae in published reports were re-examined. Their taxonomic relationships were investigated by both conventional phenotypic characterization and by DNA/DNA hybridization using the renaturation method. The human Pasteurella ureae strains were highly homogeneous in their phenotypes and in their DNA reassociation. The strain of Actinobacillus hominis studied was genetically distinct from Pasteurella ureae , but was located, like Pasteurella ureae , in the Actinobacillus group. The remaining strains exhibited only low DNA relatedness with Pasteurella ureau and each other; this agreed with their phenotypic divergence. Two of the murine isolates were identified as indole-negative variant strains of Pasteurella pneumotropica sensu stricto (i.e., type Jawetz), or of the type Heyl of Pasteurella pneumotropica , respectively. The remaining murine isolate appears to represent a hitherto unrecognized species of Pasteurellaceae. So far, there is no evidence for the occurrence of Pasteurella ureae outside the human host.

So-called Moraxella (or Pasteurella) anatipestifer and members of the Flavobacterium/Cytophaga group exhibit remarkable common features: lack of flagellation, low guanine + cytosine content of the chromosomal DNA, production of menaquinones and branched-chain fatty acids, absence of carbohydrate fermentation, and similar patterns of hydrolytic enzymes. Using the renaturation method of DNA:DNA hybridization two urease-negative European isolates and the urease-positive type strain (which was isolated in the United States) of M. P. anatipestifer were shown to have about 85% of their genome DNA base sequences in common; they may represent two subspecies. The type strain of this species was neither measurably related to the type species of the genus Moraxella nor to selected members of the family Pasteurellaceae (Pohl 1981). On the other hand, low but significant degrees of DNA binding between selected strains of so-called M. anatipestifer, Cytophaga marinoflava, Flavobacterium meningosepticum, F. odoratum and F. pectinovorum were observed. On the basis of these findings the transfer of the so-called M. anatipestifer to the Flavobacterium/Cytophaga group (family Cytophagaceae) is proposed. More detailed investigations are required to establish its relationship at the genus level.

Deoxyribonucleic acid from Legionella pneumophila showed little or no relatedness to deoxyribonucleic acids from members of the families Cytophagaceae and “Pasteurellaceae” (not on Approved Lists of Bacterial Names [Skerman et al., ed., Int. J. Syst. Bacteriol 30:225–420, 1980]) or from Kingella kingae. These data support the recent recognition of L. pneumophila as a new species.