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Experimental model of swine pneumonic pasteurellosis using crude Actinobacillus pleuropneumoniae cytotoxin and Pasteurella multocida given endobronchially.

This study was designed to develop and characterize a swine pneumonic pasteurellosis model by concurrent introduction of Pasteurella multocida type A and Actinobacillus pleuropneumoniae crude cytotoxin. After a series of preliminary experiments, a combination of 4 x 10(9) P. multocida and 4,000 toxic units of A. pleuropneumoniae crude cytotoxin was determined to produce optimal results. A total of 48 pigs were divided into four groups of 12 pigs each. The control group received buffered saline only. Four pigs from each group were randomly selected for necropsy 3, 7 and 14 days postinoculation (PI). Inoculation of pigs with P. multocida and A. pleuropneumoniae cytotoxin (group 1) resulted in moderate to severe pneumonia. Pasteurella multocida was isolated from pneumonic lesions, grossly normal lung, and bronchial lymph nodes of all group 1 pigs throughout the 14 day experimental period. Pathological changes typical of field cases of swine pneumonic pasteurellosis were produced. Pigs inoculated with P. multocida alone (group 2) had pneumonic lesions and P. multocida was reisolated from lungs at three days PI. Pasteurella multocida was not isolated from these pigs at 7 and 14 days PI, except for one pig in which an abscess developed in the thorax. Pulmonary lesions induced by A. pleuropneumoniae crude cytotoxin alone (group 3) were transient and resolved by seven days PI. Group 1 pigs had significantly greater lung lesion volumes than group 2 and 3 pigs at 3, 7 and 14 days PI. Statistical analysis indicated a significant interactive effect of P. multocida and A. pleuropneumoniae cytotoxin on the development of lung lesion volumes at 7 and 14 days PI (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Actinobacillus pleuropneumoniae↗

[Toxin-producing pasteurellas in a cattle herd].

Besides other agents, indole-negative Pasteurellae, producing dermonecrotic Pasteurella toxin, were isolated from the noses of calves in a herd with enzootic bronchopneumonia. In some blood sera, antitoxin was detected. According to their biochemical activities, isolated strains were classified as P. multocida ssp. septica (ornithine-negative), P. avium (biovar 2), and P. canis (biovar 2). However, in DNA-DNA hybridisation tests there was much coincidence with P. multocida. In experimental calves, pneumonic lesions were produced with one of the isolates as well as with the dermonecrotic toxin. Therefore, indole negative toxinogenic Pasteurellae are considered pneumonia causing agents. They should be taken into account in bacteriological diagnostic and for production of herd specific bacterins.

Animals↗

Pasteurella multocida in psittacines: prevalence, pathology, and characterization of isolates.

Although the pathogenicity of Pasteurella multocida for psittacines (parrots and their relatives) has been documented in several case reports, the associated pathologic syndromes have not been well defined nor have the isolates been characterized. In addition, the prevalence of P. multocida in psittacines has not been determined. Three hundred twenty-eight psittacines (253 clinically healthy and 75 clinically ill) were cultured for P. multocida. Pasteurella multocida was not isolated from the pharynx, choana, or cloaca of psittacines. However, in five dead psittacines submitted for necropsy, P. multocida was isolated. These isolates were characterized, and all belonged to either somatic serotype 3 or 4,7. Pasteurella multocida somatic serotype 3 was isolated from psittacines with septicemia, whereas P. multocida somatic serotype 4,7 was isolated from psittacines with cutaneous lesions. The majority (four out of five) of the P. multocida isolates belonged to the subspecies multocida, and all isolates were susceptible to penicillin G, sulfisoxazole, gentamicin, erythromycin, tetracycline, and trimethoprim-sulfamethoxazole but resistant to streptomycin. DNA fingerprints demonstrated that isolates belonging to the same somatic serotype were genetically related. The isolate from a cockatiel that had been caught by a cat belonged to somatic serotype 3 and was not genetically related to the other two isolates belonging to this somatic serotype.

Animals↗

Pasteurella infections of the head and neck.

Infections caused by Pasteurella occur most frequently after domestic animal bites or scratches and in individuals with agricultural or veterinary contact with animals. A serious Pasteurella infection developed in an agricultural worker following tumor extirpation of a head and neck neoplasm. Review of Pasteurella infections in humans disclosed that 31 of 446 reported infections involved head and neck structures. The most serious of these involved the adjacent central nervous system. Surgical drainage combined with parenteral penicillin remains the treatment of choice in these infections. Aminoglycosides are not effective in treating this organism.

Anti-Bacterial Agents↗

Fatty acid profiles of "Pasteurella" piscicida: comparison with other fish pathogenic gram-negative bacteria.

The fatty acid methyl ester (FAME) profiles of "Pasteurella" piscicida were determined by gas chromatography and subjected to numerical analysis in comparison with those obtained for Vibrio anguillarum, Aeromonas salmonicida and Pasteurella species of clinical origin. The bacterial species studied shared important characteristics with respect to their FAME content: in all of them the saturated and unsaturated fatty acids of 16 carbon atoms were the predominant fatty acids. However, distinguishing features could be detected for each pathogen. Using either single linkage or complete linkage algorithms, strains were divided into four phena that corresponded to the different species, but showed a high degree of correlation among them. Although single linkage discriminated strains better within each phenum, complete linkage was more useful to establish the relationships among clusters. The results obtained support the idea that "Pasteurella" piscicida is related to members of the genera Vibrio and Aeromonas and indicate the need for exhaustive genetic studies to clarify the taxonomic position of this fish pathogen.

Animals↗

Characterization of Pasteurella from gingival scrapings of dogs and cats.

Gingival scrapings of 62 dogs and cats were examined for the presence of Pasteurella. Isolation was performed in a medium supplemented with thiostrepton. Twenty-eight and 37 strains were obtained from 21 dogs and 26 cats, respectively, and classified in recently described species or subspecies of the genus Pasteurella (P.): P. multocida subspecies multocida and septica, P. canis, P. dagmatis and P. stomatis. Twenty-one strains were classified as atypical P. stomatis and one strain obtained from a cat remained unclassified. All strains were susceptible to the antibiotics studied. P. multocida and P. stomatis (including atypical strains) represented 65 and 30% of feline isolates, and 14 and 68% of canine isolates, respectively. Assuming that P. multocida, P. canis and P. dagmatis are potentially pathogenic for humans, and that P. stomatis has a low pathogenicity or non-pathogenic, 77 and 28% of examined cats and dogs harboured one or several pathogenic strains. This difference could explain the fact that Pasteurella infections in man are lower in dog bites rather than cat bites.

Agar↗

[Endocarditis due to Pasteurella sp. Two cases].

Human pasteurellosis is, in general, a locoregional infection due to contact with an animal. Systemic infections are rare and endocarditis is exceptionally described. The authors report two new cases of endocarditis due to Pasteurella spp, they then review 29 other published cases. Pasteurella spp. endocarditis presents as an acute form in 64% of cases and affects the aortic as often as the mitral valves. Contact with an animal is documented in 65% of cases. Pasteurella multocida is the most frequent species in this infection. The total death rate is 40% and can reach 57% of cases in case of immunodepression. The bad prognosis of this infection, justifies an early diagnosis and a rapid and adapted but not yet consensual medicosurgical treatment.

Aged↗

Differentiation between major species of the Actinobacillus--Haemophilus--Pasteurella group by gas chromatography of trifluoroacetic acid anhydride derivatives from whole-cell methanolysates.

A method based on whole-cell methanolysis and trifluoroacetic acid anhydride derivatization was developed for routine laboratory differentiation between isolates from the Actinobacillus--Haemophilus--Pasteurella group. All species, except Haemophilus aphrophilus, contained D-glycero-D-mannoheptose, although in varying concentrations. The distribution of this sugar could be used to distinguish H. aphrophilus from Actinobacillus actinomycetemcomitans, H. paraphrophilus, H. influenzae type b, Pasteurella haemolytica, P. multocida and P. ureae, and also H. influenzae type b from Pasteurellae. The pattern of major sugars in P. ureae and P. haemolytica resembled that of A. actinomycetemcomitans. Major fatty acids of the whole-cell methanolysates provided no basis of interspecies differentiation.

Acetic Anhydrides↗

Diversity of Mannheimia haemolytica and pasteurella trehalosi serotypes from apparently healthy sheep and abattoir specimens in the highlands of Wollo, North East Ethiopia.

The prevalence and serotypic diversity of Mannheimia [Pasteurella] haemolytica and Pasteurella trehalosi from nasal swabs, sera and abattoir specimens from sheep in the highlands of Wollo, North East Ethiopia was investigated. Prevalence rates of 83% and 75% of these microorganisms were found in the serum samples and nasal swabs, respectively, from apparently healthy sheep. In a local abattoir, 205 lungs were investigated, 34% of which showed pneumonia, from which samples were collected from 51 lungs and the same number of corresponding tonsils. Mannheimia and Pasteurella species were isolated from 59% of these pneumonic lungs and 69% of the respective tonsils. M. haemolytica serotypes accounted for 41 (59%) and P. trehalosi for 11 (32%) of the isolates from the abattoir specimens. The majority (67%) of isolates from nasal swabs were P. trehalosi, M. haemolytica being isolated f rom 4 (13%) of the swabs. M. glucosida was isolated only from the tonsils. The predominant serotypes of the isolates from both the nasal swabs and the abattoir specimens were M. haemolytica A1 (17%) and P. trehalosi T4 (16%) and T3 (13%). P. trehalosi T15 was less commonly encountered, while M. haemolytica A9 and A13 were not isolated. Studies on sera from 100 sheep indicated that antibodies against M. haemolytica serotype A1 (14%) were most common, followed by A5 and A8 (each 10%) and A9 and P. trehalosi T3 (each 9%) and T4 (8%). Antibodies against M. glucosida or serotype All occurred in 2% of the sera. Multiple serotypes were common in all types of samples. The importance of including in vaccines the most prevalent serotypes involved in the pneumonia of sheep in the area is discussed.

Animals↗

Genotypic heterogeneity of Pasteurella gallinarum as shown by ribotyping and 16S rRNA sequencing.

Forty-five strains mainly isolated from chickens in Zimbabwe and Denmark, two pig and three rat isolates all identified as Pasteurella gallinarum by conventional phenotypic tests were characterized by ribotyping, and selected strains were subsequently analysed by 16S rRNA gene sequencing. High genotypic diversity was observed, the number of ribotypes totalling 24. A major group of 47 isolates including the type strain of P. gallinarum clustered at 56% similarity and included 21 ribotypes. Ribotyping showed that some genotypes of P. gallinarum seem to be globally distributed. The three isolates from rodents did not share even a single common ribotype fragment with strains from birds and the pig isolates. Two avian isolates from Denmark and Zimbabwe and the pig strain showed from 97.6 to 99.8% 16S rRNA sequence similarity with the type strain of P. gallinarum and with type strains of Pasteurella volantium and Pasteurella avium. Two rat strains showed 98.6% 16S rRNA gene sequence similarity with each other, but were only related with P. gallinarum at 93% similarity. These isolates showed the highest similarity with [Actinobacillus] muris at 96.4 to 95.0% similarity. We suggest that conventional identification of P. gallinarum consequently should consider the source of isolation to obtain a correct diagnosis, and that isolation from animals other than fowl should be confirmed by genotypic analysis such as 16S rRNA gene sequence comparison.

Animals↗

Small-subunit rRNA sequences and whole DNA relatedness concur for the reassignment of Pasteurella piscicida (Snieszko et al.) Janssen and Surgalla to the genus Photobacterium as Photobacterium damsela subsp. piscicida comb. nov.

The taxonomic status of Pasteurella piscicida (strain NCIMB 2058T [T = type strain] and a strain isolated from the environment) was investigated by performing phylogenetic analyses of small-subunit rRNA sequences, DNA-DNA hybridization analyses, and biochemical characterization analyses. The results of the phylogenetic analyses and the levels of DNA-DNA complementarity demonstrated conclusively that Pasteurella piscicida is extremely closely related to Photobacterium damsela ATCC 33539T. Since the two taxa exhibited a level of DNA-DNA relatedness of 80%, they are members of the same species. The high level of DNA relatedness and the presence of specific morphological and biochemical characteristics support the hypothesis that two subspecies should be recognized. On the basis of its phylogenetic position, we concluded that Pasteurella piscicida should be renamed Photobacterium damsela subsp. piscicida comb. nov.

Base Sequence↗

A numerical taxonomic study of Actinobacillus, Pasteurella and Yersinia.

A numerical taxonomic study of strains of Actinobacillus, Pasteurella and Yersinia, with some allied bacteria, showed 23 reasonably distinct groups. These fell into three major areas. Area A contained species of Actinobacillus and Pasteurella: A. suis, A. equuli, A. lignieresii, P. haemolytica biovar A, P. haemolytica biovar T, P. multocida, A. actinomycetemcomitans, 'P. bettii', 'A. seminis', P. ureae and P. aerogenes. Also included in A was a composite group of Pasteurella pneumotropica and P. gallinarum, together with unnamed groups referred to as 'BLG', 'Mair', 'Ross' and 'aer-2'. Area B contained species of Yersinia: Y. enterocolitica, Y. pseudotuberculosis, Y. pestis and a group 'ent-b' similar to Y. enterocolitica. Area C contained non-fermenting strains: Y. philomiragia, Moraxella anatipestifer and a miscellaneous group 'past-b'. There were also a small number of unnamed single strains.

Actinobacillus↗

Multivariate analysis of quantitative chemical and enzymic characterization data in classification of Actinobacillus, Haemophilus and Pasteurella spp.

Chemotaxonomic data for strains of Actinobacillus, Haemophilus and Pasteurella spp. were analysed using three multivariate statistical strategies: principal components, partial least squares discriminant, and soft independent modelling of class analogy. The species comprised Actinobacillus actinomycetemcomitans. Haemophilus aphrophilus, H. paraphrophilus, H. influenzae, Pasteurella multocida, P. haemolytica and P. ureae. Strains were characterized by cell sugar and fatty acid composition, lysis kinetics during EDTA and EDTA plus lysozyme treatment, and methylene blue reduction. In total 23 quantitative variables were compiled from chemotaxonomic analyses of 25 strains. A. actinomycetemcomitans and H. aphrophilus formed distinct classes which differed from those of H. paraphrophilus, H. influenzae and Pasteurella spp. All characterization variables, except those describing fatty acid content, contributed significantly to inter-species discrimination.

Actinobacillus↗

Effects of experimentally induced Pasteurella haemolytica infection in dairy calves on the pharmacokinetics of flumequine.

The effect of experimental Pasteurella haemolytica infection on the intravenous and intramuscular pharmacokinetics of flumequine was studied in dairy calves. The plasma concentration-time curve of flumequine after intravenous injection of 5 mg/kg bodyweight flumequine of a 10% solution before and after experimental infection, was best described by a three-compartment open model. After intramuscular injection of the same dosage rate of a 3% flumequine suspension is was best described by the one-compartment open model with first-order absorption. The experimental infection by intratracheal administration of infectious bovine rhinotracheitis (IBR)-virus and 5 days later intrapulmonary administration of Pasteurella haemolytica produced a clear temperature rise and signs of disease expressed as Average Health Status. Subsequently, plasma Fe and Zn concentration decreased after infection. The distribution volumes Vc, Vd(area) and Vd(ss) after infection (0.07 +/- 0.04, 1.38 +/- 0.36 and 0.50 +/- 0.11 l/kg, respectively) were smaller than those before infection, but the differences were not significant (P less than or equal to 0.1). The intravenous AUC infinity was significantly increased (21.86 +/- 3.51 to 33.85 +/- 2.97 mg.h/l, P less than or equal to 0.01) and the total body clearance (ClB) significantly decreased (0.24 +/- 0.02 to 0.15 +/- 0.01, P less than or equal to 0.01) after infection. After intramuscular injection of flumequine at 5 mg/kg as a 3% suspension, only the bioavailability, F, was significantly decreased after infection (78.5 +/- 14.3 to 59.7 +/- 21.2%, P less than or equal to 0.02). However, this had no consequences for the dosage regimen used. The urine concentration ratio flumequine:7-hydroxy-flumequine:conjugated flumequine changed from 2:1:10 before infection to 6:1:15 after infection, which indicates that hydroxylation and glucuronidation as metabolic pathways for flumequine were decreased after Pasteurella sp. infection.

Animals↗

Cloning of the gene and characterization of the enzymatic properties of the monomeric alkaline phosphatase (PhoX) from Pasteurella multocida strain X-73.

We have identified a new phoX gene encoding the monomeric alkaline phosphatase from Pasteurella multocida X-73. This gene was not found in the published genome sequence of Pasteurella multocida pm70. Characterization of the recombinant PhoX of Pasteurella multocida X-73 showed that it is a monomeric enzyme, activated by Ca(2+) and possibly secreted by the Tat pathway. These features distinguish phosphatases of the PhoX family from those of the PhoA family. All proteins of the PhoX family were found to contain a conserved motif that shares significant sequence homology with the calcium-binding site of a phosphotriesterase known as diisopropylfluorophosphatase. Site-directed mutagenesis revealed that D527 of PhoX might be the ligand bound to the catalytic calcium. This is the first report on identification of homologous sequences between PhoX and the phosphotriesterase and on the potential calcium-binding site of PhoX.

Alkaline Phosphatase↗

Characterization of some previously unclassified "Pasteurella" spp. obtained from the oral cavity of dogs and cats and description of a new species tentatively classified with the family Pasteurellaceae Pohl 1981 and provisionally called taxon 16.

The taxonomic relationship of 23 unclassified canine and feline strains of Pasteurellaceae and five strains received as Pasteurella spp. or Haemophilus influenzae-murium was investigated by phenotypic and genetic characterization. Eleven strains were classified with four recently described species of genus Pasteurella sensu stricto. Fourteen canine and feline strains formed a homogeneous group, tentatively designated taxon 16. Both phenotypic characters and mol% G + C in DNA and genome size indicate classification of taxon 16 with genus Pasteurella. DNA/DNA hybridizations, however, failed to locate taxon 16 on genus level with accepted or proposed species of the family Pasteurellaceae Pohl 1981. Two additional species obtained from rats and mice remained unclassified, in addition to a human isolate obtained from a dog-bite. The necessity of detailed phenotypic characterization within the family Pasteurellaceae Pohl 1981 needs to be stressed.

Animals↗

Synergistic effect in viral-bacterial infection: combined infection of the murine respiratory tract with Sendai virus and Pasteurella pneumotropica.

Synergism was demonstrated between Sendai virus and Pasteurella pneumotropica in the respiratory tract of mice showing no evidence of previous infection with either agent. Mice aerosol challenged with P. pneumotropica invariably eliminated the viable organism from their lungs within 72 h. In contrast, intrapulmonary killing was delayed in animals previously infected with Sendai virus. Maximum synergism was observed when virus infection preceded bacterial challenge by 6 days. At this time, a mortality rate of 37% was observed as compared with 0, 10, 20, and 10%, respectively, in those animals in which the virus infection preceded bacterial challenge by 1, 3, 9, and 12 days. Previous immunization with Sendai virus completely prevented virus infection and thus the synergistic effect. Synergism with endogenous flora was also noted. Six days after virus infection an endogenous Pasteurella sp. began to proliferate in the bronchopulmonary tissues. Up to 10(4) colony-forming units per lung were recovered but no animals died of the endogenous Pasteurella infection.

Aerosols↗

Metabolism of carbohydrates by Pasteurella pseudotuberculosis.

Cell-free extracts of Pasteurella pseudotuberculosis and P. pestis catalyzed a rapid and reversible exchange of electrons between pyridine nucleotides. Although the extent of this exchange approximated that promoted by the soluble nicotinamide adenine dinucleotide (phosphate) transhydrogenase of Pseudomonas fluorescens, the reaction in the pasteurellae was associated with a particulate fraction and was not influenced by adenosine-2'-monophosphate. The ability of P. pseudotuberculosis to utilize this system for the maintenance of a large pool of nicotinamide adenine dinucleotide phosphate could not be correlated with significant participation of the Entner-Doudoroff path or catabolic use of the hexose-monophosphate path during metabolism of glucose. As judged by the distribution of radioactivity in metabolic pyruvate, glucose and gluconate were fermented via the Embden-Meyerhof and Entner-Doudoroff paths, respectively. With the exception of hexosediphosphatase, all enzymes of the three paths were detected, although little or no gluconokinase or phosphogluconate dehydrase was present unless the organisms were cultivated with gluconate. The significance of these findings is discussed with respect to the regulation of carbohydrate metabolism in the pasteurellae, related enteric bacteria, and P. fluorescens.

Carbon Isotopes↗