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Taxonomic subgroups of Pasteurella multocida correlate with clinical presentation.

Pasteurella multocida is a small nonmotile gram-negative coccobacillus that is found in the nasopharynx and gastrointestinal tract of many wild and domesticated animals. In humans it most commonly causes cellulitis and localized superficial skin abscesses following an animal bite or scratch. The respiratory tract is the second most common site of infection for PASTEURELLA: Of the more than 17 species of Pasteurella known, Pasteurella multocida subsp. multocida and Pasteurella multocida subsp. septica are among the most common pathogens in humans. With the use of molecular techniques, distinction between different subspecies of P. multocida can be made more easily and accurately. We used the sequence of the 16S ribosomal DNA (rDNA) and repetitive extragenic palindromic sequence-PCR (REP-PCR) to characterize 20 strains (14 of P. multocida subsp. multocida and 6 of P. multocida subsp. septica; the 16S rDNA is identical for P. multocida subsp. multocida and Pasteurella multocida subsp. gallicida but differs from that of P. multocida subsp. septica) isolated from various anatomic sites. We found excellent correlation between the 16S rDNA sequence (a marker for a small conserved region of the genome), REP-PCR (a marker for a large portion of the genome), and biochemical tests (trehalose and sorbitol). We also found a correlation between the clinical presentation and the taxonomic group, with P. multocida subsp. septica more often associated with wounds than with respiratory infections (67 versus 17%, respectively) (P < 0.05, Z test) and P. multocida subsp. multocida more often associated with respiratory infections than with wounds (71 versus 14%, respectively) (P < 0.05, Z test).

Bacterial Typing Techniques↗

[Clinicobacteriological study of Pasteurella multocida infection as a zoonosis. (2)--Comparisons of P. multocida separated from patients and their pet dogs and cats].

A total of 17 strains of Pasteurella multocida, of which 13 were isolated from patients treated at Nihon University Itabashi Hospital or Nihon University Surugadai Hospital between April, 1984 and March, 1991 and 4 from 1 dog and 3 cats kept by the patients, were evaluated with respect to their biochemical properties, sensitivity to drugs, and serotype. The isolated strains were all considered to be Pasteurella multocida subsp. multocida because of the agreement of their responses to indole, sorbitol and dulcitol with those of this subspecies, except for 1 sorbitol-negative strain of Pasteurella multocida subsp. septica isolated from 1 patient who had been bitten by a cat. All the isolated strains showed high sensitivities to various drugs. The serotype was capsular type A, which is often observed in cats and dogs, in 7 strains, which consisted of 6 of the 7 strains derived from the airway of the patients and 1 of the 6 strains derived from bit or scratch wound. The remaining strains could not be classified. Five morphological types, namely 1, 3, 3.8, 6, and 8 were observed. In 2 patients, Pasteurella multocida subsp. multocida of the same serotype was also isolated from their cats. One of these patients had intimate contact with the cat including kissing. Our findings suggest that: 1) Pasteurella multocida subsp. multocida has been responsible for most conventional cases of Pasteurella multocida infection. 2) Strains isolated from patients differ in the capsular type according to the disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Identification of Pasteurella serotypes by indirect hemagglutination].

The indirect haemagglutination test was used for serological type identification of Pasteurella strains. The Pasteurella multocida strains were tested by parallel application of antigen production methods (common method plus hyaluronidase treatment). Thirty-nine (72.2 per cent) of 54 Pasteurella multocida strains, which had been obtained from cattle and swine, were serologically tested by the common technique of antigen production, their types being identified, while 42 (77.7 per cent) were tested and typed by means of hyaluronidase treatment. Depolimerisation of hyaluronic acid by means of hyaluronidase increased the agglutinability of eight strains (14.8 per cent) which could thus be identified by types. Serological identification of 47 (87.1 per cent) Pasteurella multocida strains was achieved by parallel use of the two antigen production methods. Identification was not possible of two strains. Cross-reaction was recorded from five strains. Most of the Pasteurella multocida strains isolated from cattle were A serotype (54 per cent), while those of swine were B strains (50 per cent). Thirty-six of 40 serologically tested Pasteurella haemolytica strains were serotypes A1, A2, T3, T4, A5, and A6, and most of these were A5 and T4.

Animals↗

Pasteurella multocida pneumonia.

Pasteurella multocida, a gram-negative coccobacillus which colonizes the nasopharynx and gastrointestinal tract of many animals, is a well known cause of soft tissue infection after animal bites. Human infection can also occur after non-bite animal exposure, usually via inhalation of contaminated secretions. The respiratory tract is the second most common site of Pasteurella infection after soft tissue infection. Most patients with Pasteurella pulmonary infection are elderly with underlying lung disease, either COPD, bronchiectasis, or malignancy. The spectrum of disease includes pneumonia, tracheobronchitis, lung abscess, and empyema. Clinical features of Pasteurella respiratory tract infections are indistinguishable from other pathogens. A history of cat or dog exposure should alert the clinician to consider Pasteurella as a potential pulmonary pathogen in an elderly patient with chronic lung disease. The preferred drug for the treatment of Pasteurella infections is penicillin. Alternately, doxycycline is highly effective against P multocida.

Animals↗

Occurrence of sialidase and N-acetylneuraminate lyase in Pasteurella species.

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.

Animals↗

Taxonomy of pasteurella anatipestifer. 1. DNA base composition and DNA-DNA hybridization analysis.

DNA was isolated from 15 strains of Pasteurella anatipestifer and from one strain each of Moraxella nonliquefaciens, M. bovis, Pasteurella multocida, P. haemolytica, P. gallinarum, P. pneumotropica, and P. ureae. The guanine-plus-cytosine contents of P. anatipestifer ranged from 32 to 35 mole %, whereas those of Moraxella and Pasteurella spp. were much higher, ranging from 40 to 45 mole %. DNA-DNA hybridization analysis revealed that homology of nine P. anatipestifer strains to strains ATCC 11845 and PA 15 was 52 to 100%, whereas homology of Moraxella and Pasteurella strains to these strains was only 3 to 17%. Similarly, homology of P. anatipestifer strains, Moraxella, and Pasteurella species other than P. multocida to P. multocida reference strain P-2192 was low. These results strongly suggest that P. anatipestifer is genetically unrelated to either Pasteurella or Moraxella.

Base Composition↗

Increased frequency of isolation of Pasteurella and Actinobacillus species and related organisms.

Ninety-six clinical isolates of Pasteurella, Actinobacillus and related organisms were submitted to our reference laboratory for identification. The procedures for detecting the 11 identified species, Pasteurella multocida, Pasteurella haemolytica, Pasteurella ureae, Pasteurella pneumotropica, Actinobacillus lignieresii, Actinobacillus equuli, Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Cardiobacterium hominis, and two unnamed species of CDC groups, HB-5 and EF-4, are described and their clinical importance is discussed. These organisms have been increasingly isolated in Japan and are most often associated with respiratory infections and endocarditis.

Actinobacillus↗

Ultrastructural pathology of the upper respiratory tract of rabbits experimentally infected with Pasteurella multocida A:3.

Twenty-four 8 to 9 week-old Pasteurella multocida -free rabbits were divided into three equal groups, the first group was pretreated with hydrocortisone and inoculated intranasally with pasteurella multocida serotype A:3. The second group was inoculated intranasally with P. multocida without hydrocortisone treatment. The third group was inoculated with phosphate buffered saline only and used as a control group. Pasteurella multocida was isolated from the nasal cavity of all infected rabbits in group 1 and 2 and from the trachea of seven rabbits in group 1 and five rabbits in group 2. This study was conducted to observe the ultrastructural changes of the upper respiratory tract of hydrocortisone treated and non-treated rabbits infected with P. multocida serotype A:3. The ultrastructural changes detected in infected rabbits were ciliary destruction and deciliation of the ciliated epithelial cells, cellular swelling, goblet cell hyperplasia and endothelial cell damage. Pasteurella multocida was observed attached to the degenerated cilia, microvilli and mucus. Pasteurella multocida infection was associated with inflammatory responses, which may have caused tissue damage. It is possible that hydrocortisone modulates the severity of infection as an immune suppressor and an inhibitor of goblet cell secretion.

Animals↗

Brain abscess due to Pasteurella multocida.

The patient was a 26-year-old man who complained of headache and vomiting. On examination, there was nothing abnormal, but the edge of the right optic papilla was not clear. His temperature was 38.5 degrees C, pulse 96/min, blood pressure 120/80 mmHg. A space-occupying lesion in his fronto-dextra cupular part was found by CT scanning. He had a 12-year history of chronic purulent otitis. The diagnosis was a brain abscess in the fronto-dextra cupular part. The brain abscess was extracted and Pasteurella multocida was isolated from the dark brown pus draining from the abscess. The patient recovered through proper antibiotic therapy based on a sensitivity test. Reports of infections caused by this organism in foreign countries very widely from local infections due to bites and scratches by cats, dogs etc. to general infections such as infections of the respiratory tract, sepsis and meningitis. However, Pasteurella multocida brain abscesses are rare. Pasteurella multocida is a Gram-negative short rod which is best known as part of the mouth flora and as a pathogen causing septicemia in many domestic animals, such as cats, dogs etc.. Infection in man results mainly from animal bites or scratches. It has been reported that Pasteurella multodida can cause human septicemia, meningitis, respiratory tract infection, conjunctivitis and other infections. We isolated a strain of Pasteurella multocida from the pus of a brain abscess following chronic purulent otitis on August 6, 1990.

Adult↗

Characteristics and biotypes of Pasteurella multocida isolated from humans.

Fifty-two isolates of Pasteurella (48 strains of Pasteurella multocida and 4 strains of atypical Pasteurella) were identified by conventional and commercial test systems. All strains fermented glucose, sucrose, and fructose in purple broth base (Difco Laboratories) with bromocresol purple as indicator, although the atypical Pasteurella produced fermentation reactions that were barely perceptible. Eleven different biotypes were identified by fermentation reactions in maltose, mannitol, xylose, sorbitol, and trehalose media. There was a correlation of biotypes to cat bites, with 61% of cat bite isolates falling into biotype A and B. A correlation of biotype and dog bite isolates was not seen. The choice of medium used for fermentation tests was critical as evidenced by the inability of the organisms to grow in a second commercially purchased preparation of purple broth base. The reliability of commercial test systems in identifying Pasteurella was 81% for Oxi/Ferm (Roche Diagnostics, Div. Hoffmann-La Roche, Inc., Nutley, N.J.), 68% for API (Analytab Products, Plainview, N.Y.), and 11% for Minitek (BBL Microbiology Systems, Cockeysville, MD.).

Animals↗

Isolation of Pasteurella spp. from free-ranging American bison (Bison bison).

From November 1991 through March 1992, nasal and pharyngeal swab samples were collected from 45 bison (Bison bison) from Yellowstone National Park, Montana (USA) and cultured for Pasteurella spp. Thirteen isolates of Pasteurella spp. were recovered from 10 (22%) of the animals. Ten isolates were from pharyngeal samples in contrast to three isolates from nasal samples. Pasteurella haemolytica (six biotype T, two biotype A, and two biotype 3) was the predominant Pasteurella species. Five biotype T isolates were serotype 4 and the sixth agglutinated in antisera 3, 4, and 10. Both biotype A isolates were untypable with antisera to recognized type strains. Pasteurella multocida was isolated from the pharyngeal samples of one animal. Two isolates could not be identified to species.

Animals↗

Sharing of Pasteurella spp. between free-ranging bighorn sheep and feral goats.

Pasteurella spp. were isolated from feral goats and free-ranging bighorn sheep (Ovis canadensis canadensis) in the Hells Canyon National Recreation Area bordering Idaho, Oregon, and Washington (USA). Biovariant 1 Pasteurella haemolytica organisms were isolated from one goat and one of two bighorn sheep found in close association. Both isolates produced leukotoxin and had identical electrophoretic patterns of DNA fragments following cutting with restriction endonuclease HaeIII. Similarly Pasteurella multocida multocida a isolates cultured from the goat and one of the bighorn sheep had D type capsules, serotype 4 somatic antigens, produced dermonecrotoxin and had identical HaeIII electrophoretic profiles. A biovariant U(beta) P.haemolytica strain isolated from two other feral goats, not known to have been closely associated with bighorn sheep, did not produce leukotoxin but had biochemical utilization and HaeIII electrophoretic profiles identical to those of isolates from bighorn sheep. It was concluded that identical Pasteurella strains were shared by the goats and bighorn sheep. Although the direction of transmission could not be established, evidence suggests transmission of strains from goats to bighorn sheep. Goats may serve as a reservoir of Pasteurella strains that may be virulent in bighorn sheep; therefore, goats in bighorn sheep habitat should be managed to prevent contact with bighorn sheep. Bighorn sheep which have nose-to-nose contact with goats should be removed from the habitat.

Animals↗

Studies on the occurrence of Gram-negative bacteria in ticks: Ixodes ricinus as a potential vector of Pasteurella.

A total of 372 Ixodes ricinus ticks (101 females, 122 males, and 149 nymphs) collected by flagging in 6 mixed woodlands of eastern Poland were examined by culture for the presence of internal Gram-negative bacteria other than Borrelia burgdorferi. Adult ticks were examined in pools of 2 specimens each and nymphs were examined in pools of 3-5 specimens each. Ticks were disinfected in 70 % ethanol and homogenized in 0.85% NaCl. The diluted homogenate was inoculated onto 3 kinds of agar media: buffered charcoal yeast extract (BCYE-alpha) for isolation of fastidious Gram-negative bacteria, eosin methylene blue agar (EMB) for isolation of enterobacteria, and tryptic soya agar for isolation of all other non-fastidious Gram-negative bacteria. The Gram-negative isolates were identified with the API Systems 20E and NE microtests. A total of 9 species of Gram-negative bacteria were identified, of which the commonest were strains determined as Pasteurella pneumotropica/haemolytica, which were isolated on BCYE-alpha agar from ticks collected in all 6 examined woodlands. The total number of these strains (49) exceeded the total number of all other strains of Gram-negative bacteria recovered from ticks (30). Of the total number of examined ticks, the minimum infection rate with Pasteurella pneumotropica/haemolytica was highest in females (18.8%), and slightly lower in males (12.3%) and nymphs (10%). Besides Pasteurella pneumotropica/haemolytica, the following species of Gram-negative bacteria were isolated from examined ticks: Pantoea agglomerans, Serratia marcescens, Serratia plymuthica on EMB agar and Aeromonas hydrophila, Burkholderia cepacia, Chromobacterium violaceum, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia on tryptic soya agar. Minimal infection rates with these bacteria were low, ranging from 0.7-5.9%. Of the isolated bacteria, Chromobacterium violaceum, Pasteurella pneumotropica/haemolytica, Pseudomonas aeruginosa, and Serratia marcescens are potentially pathogenic for man and/or animals. In particular, the common occurrence of Pasteurella pneumotropica/haemolytica in Ixodes ricinus ticks poses a potential risk of pasteurellosis for humans and animals exposed to tick bites.

Animals↗

Demonstration of Pasteurella-specific immunoglobulin E in bovine serum.

On the basis of recent observations that immunoglobulin (Ig) E antibodies specific for bacterial antigens occur in the serum of persons with chronic respiratory tract disease, we used bovine epsilon chain-specific antiserum to investigate the possibility that IgE antibodies are induced in cattle infected with Pasteurella. Using enzyme-linked immunosorbent assay and Western blotting techniques, we studied bovine sera to detect and quantitate the presence of IgE antibodies specific for antigens of Pasteurella. Immunoglobulin E antibodies reactive with whole formalinized P haemolytica, potassium thicyanate, and saline solution extracts were detected in serum of calves with bronchopneumonia, feedlot steers with interstitial pneumonia, as well as nonaffected penmates, and adult dairy cows. The role of parenteral vaccination in eliciting an IgE response was examined in healthy calves; vaccination with a Pasteurella bacterin failed to induce an IgE response. Adsorption studies were done to demonstrate the specificity of the antibodies for Pasteurella. Enzyme-linked immunosorbent assay absorbance values were significantly decreased by adsorption with P haemolytica, whereas adsorption with other gram-negative bacteria only moderately decreased serum absorbance values. To begin identification of the antigen(s) to which the IgE binds, Western blotting of P haemolytica extract with sera from calves with bronchopneumonia was done. A dense band of protein (approximately 60,000 daltons) reacted strongly with IgE in the highest titer sera. These results indicate that Pasteurella-specific IgE antibodies are not readily induced by parenteral vaccination, but can be found in serum of some cattle, possibly induced by existing or previous infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Pathogenicity of a serotype 12:A Pasteurella multocida in hydrocortisone treated and nontreated rabbits.

A Pasteurella multocida isolate of 12:A serotype from a rabbit caused typical pulmonary pasteurellosis and death in pasteurella-free rabbits by intranasal exposure. Rabbits stressed with hydrocortisone and inoculated with 12:A Pasteurella multocida organisms developed a higher prevalence of pneumonia than rabbits not treated with hydrocortisone. Typical 12:A Pasteurella multocida was isolated from nasal cavity, trachea, and lungs and was most prevalent in nasal cavities. Surviving rabbits developed serum agar gel precipitating antibody beginning 15 days post-inoculation. The data showed that the 12:A Pasteurella multocida isolate was pathogenic, caused mortality, colonized the respiratory tract, and stimulated systemic immune response by producing serum agar gel precipitating antibody.

Animals↗

[Aerogenous immunostimulation of weaner swine against experimentally induced respiratory infections with Pasteurella multocida].

The clearance of Pasteurella, Bordetella and Haemophilus from the lung of weaners was increased by aerogenous immunostimulations using a lysate mixture consisting of 12 strains of 8 bacterial species, as well as live and inactivated temperature sensitive mutants of Pasteurella multocida. The severity of experimental pneumonias caused by Pasteurella has also been decreased by this treatment. The effect of the lysate was shown to be unspecific and short, as antibody production against Pasteurella was not induced and no booster effect of subsequent aerogenous immunizations with Pasteurella antigens could be detected.

Animals↗

PASTEURELLA SEPTICA INFECTIONS IN HUMANS.

Five instances of infection by Pasteurella septica, an organism which closely resembles H. influenzae on methods of presumptive bacteriology, are reported from an acute treatment general hospital practice over a period of three months. This infection decidedly outnumbers those caused by other Pasteurella species in North America. The organism is apt to be identified as H. influenzae, Friedlander's bacillus or Mima polymorpha. Unlike these, it shows extreme sensitivity to penicillin, sometimes combined with resistance to antibiotics that inhibit or kill these other organisms.While in the past Pasteurella septica infection was considered to be characterized by persistent local infection following domestic animal scratches or bites, it is clear that it is more often found in cases of respiratory tract infection and peripheral septic disease. In three of the five cases reported, exposure to animals was the source of infection.Because mixed infection with confusingly similar organisms may occur, lack of awareness of extreme penicillin sensitivity of Pasteurella septica may account for persistent failure of antibiotic therapy.

Animals↗

Further characterization of Pasteurella haemolytica-like bacteria isolated from swine enteritis.

DNA-DNA hybridization studies were conducted on six Pasteurella haemolytica-like (PHL) organisms recovered from cases of swine enteritis. Chromosomal-enriched fractions of PHL organisms served as the source of DNA for Southern blots or as whole-chromosomal DNA probes. Under stringent hybridization conditions, chromosomal DNA probes of a prototype PHL (strain 6213A) organism distinguished other PHL organisms from Pasteurella haemolytica types A1 and T3, Pasteurella multiocida types A:1 and A:3, Escherichia coli, Pseudomonas aeruginosa, Actinobacillus pleuropneumoniae type 1, and Salmonella cholerasuis. The guanine-cytosine content of the DNA of three PHL strains was 41.2 to 42.8 mol % as calculated from the thermal denaturation midpoint temperatures. The PHL strains are Gram-negative, nonmotile, beta-hemolytic, pleomorphic, oxidase-positive, urease- and indole-negative, fermentative rods with the key characteristics of the species Pasteurella haemolytica. None of the PHL strains reacted with the type-specific antisera of P. haemolytica types 1 through 12 as tested by an agglutination procedure. These swine strains differed in their biochemical differentiation from P. haemolytica types A1 and T3 in that all produced acid from M-inositol and failed to grow on MacConkey agar. Acid production from trehalose and L-arabinose was variable with PHL strains. Leukotoxicity of PHL strains was evaluated by a colorimetric micro-titration assay. Sterile culture supernatants of three of five PHL strains were toxic to bovine neutrophils. Results of these studies suggest that the PHL organisms may belong to a new group of organisms under the genus Pasteurella.

Animals↗