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Ribotype diversity of porcine Pasteurella multocida from Australia.

OBJECTIVE: To use the technique of ribotyping to investigate the genetic diversity of Australian isolates of Pasteurella multocida associated with outbreaks of clinical disease in Australian pigs. DESIGN: One hundred and seven porcine P multocida isolates were analysed by ribotyping using the restriction enzymes HpaII and HindIII. The genetic population structure of the Australian porcine P multocida isolates was determined through statistical analysis of the joint ribotype patterns, and this was then compared with biochemical and epidemiological data available for the population. RESULTS: A total of 25 combined ribotypes were recognised, which were grouped into five ribotype clusters. Despite the deliberate selection of diverse isolates, the study revealed only a limited degree of genetic diversity. Fourteen of the ribotypes contained multiple isolates, and 12 of these ribotypes were present on more than one farm. Three of the seven biovars analysed in the study showed very limited diversity. All fifteen biovar 2 isolates (subsp multocida) were found in a single cluster (III), while all four biovar 8 isolates, which correspond to P multocida subsp gallicida, were allocated by themselves to a single cluster (IV). All nine of the biovar 12 isolates (lactose-positive subsp multocida) were assigned to a single cluster (I), together with the single biovar 14 isolate, which was the only other lactose-positive isolate in the population (ODC-negative). CONCLUSION: A limited number of ribotypes of P multocida are associated with Australian pigs. The majority of these ribotypes are widely distributed across multiple farms, and across multiple states. Individual farms can possess multiple ribotypes of P multocida. Some of the unusual biochemical variants of P multocida present in Australian pigs have a very limited genetic diversity. The nature of pig production in Australia, primarily involving continuous flow systems with few closed herds, has possibly contributed to the widespread distribution of a limited number ribotypes.

Animals↗

Fever and survival in rabbits infected with Pasteurella multocida.

1. Fever and survival rate of New Zealand white rabbits, injected with two doses of live Pasteurella multocida, were compared to determine if the relation between fever and survival observed in reptiles is also seen in a mammal. Since it is known that fever is beneficial in infected reptiles, our experiments were viewed as an initial step in the investigation of a similar potentially beneficial effect in mammals.2. There was a statistically significant correlation between the fever magnitude and survival. As fever increased up to 2.25 degrees C, the survival rate increased. Fevers above this level were associated with a decreased survival rate.3. Antipyretic drugs were administered to half the rabbits. The drugs were ineffective in reducing the fevers produced by injections of large doses of bacteria. Rabbits infused with antipyretics had a decreased mortality rate. The decreased mortality rate may be due to some action of the drugs other than their antipyretic effect.4. In vitro growth of P. multocida at normal (39 degrees C) and febrile (42 degrees C) temperatures was not significantly different. This suggests that the increased survival rate associated with higher fevers was not due to a direct inhibition of the growth of the bacteria by the increased temperature. It may be due to an enhancement of some aspect of the rabbits' immunological defences.5. It remains to be determined whether fever causes a higher survival rate or whether fever is only correlated with survival rate.

Animals↗

[Human respiratory tract infection by Pasteurella multocida subsp. multocida presumably derived from the cat].

Pasteurella multocida is a Gram-negative short rod-shaped bacteria that has been recognized as a pathogen of hemorrhagic septicemia and fowl cholera in the veterinary medicine. Infections by this microorganism as seen in the foreign literature vary widely from local infections due to bites and scratches by animals to general infections such as infections of the respiratory tract, sepsis, and meningitis. In Japan, reported cases of P. multocida infections are predominantly local infections, followed by respiratory infection. Recently, death of diabetic patients due to septicemia by this pathogen has also been reported. In this study, we experienced a case of respiratory tract infection in which the pathogen P. multocida subsp. multocida was suggested to have been transmitted from a pet cat by the agreement of the serotype of the bacterial isolates between the patient and the cat. This case was evaluated from the zoonotic viewpoint. The patient was a 68-year-old male who had been followed up since 1982 with a diagnosis of bronchiectasis. After his referral to our hospital, P. multocida subsp. multocida was isolated from his bloody sputum and, then, from the cat kept by the patient. The tow isolates were identical in terms of the biochemical properties, drug susceptibility profile, and serotype (-:1), and the derivation of P. multocida subsp. multocida infection from cat was established for the first time in this report. The incidence of P. multocida infections is increasing in Japan, and particular attention is considered to be needed about these conditions as zoonoses as indicated in "Preventive Measures against Zoonoses Derived from Pet Animals (Dog, Cat)", an official communication from the Ministry of Health and Welfare to related institutions in 1989. Also, to check whether the patient keeps any pet at the clinical inquiry is a practice of bacteriological importance in all fields of medicine.

Aged↗

[Five cases of respiratory infection due to Pasteurella multocida].

Five cases of respiratory infection in which Pasteurella multocida (P. multocida) was isolated from sputum were reported. All of the five patients had animal contact, and three of the five patients suffered from bronchiectasis. No underlying pulmonary disease was found in one case, P. multocida was also isolated from her cat. Patients were treated with ST, MINO + PIPC, ABPC + CFT, SBTPC, AMK + MINO with improvement. Thirty cases of respiratory tract infection due to P. multocida including our cases have been reported in Japan.

Aged↗

[Three cases of Pasteurella multocida infection in the respiratory tract].

Pasteurella multocida (P. multocida) is well recognized as "normal flora" in the upper respiratory tract of cats, dogs and other animals. Recently, various infections due to P. multocida in human have been noted as pulmonary infections in the patients with chronic pulmonary diseases as well as skin abscesses or septicemia after an animal bite or scratch. We report here three cases of respiratory tract infections caused by P. multocida. The first two patients had acute exacerbation of bronchiectasis caused by P. multocida and the other patients with pulmonary emphysema developed pneumonia. These three patients improved by antibiotic therapy. In Japan, P. multocida respiratory tract infection is rare, but it may become more common in the future. Therefore, it seems to be important to take this pathogen into consideration in the management of chronic lung disease.

Adult↗

[A clinical study on patients detected Pasteurella multocida from sputum].

We reported ten cases, (four female and six male), whose sputum cultures positive for Pasteurella multocida from 1990 to 2000. In the past eleven years increasing numbers of cases have appeared in our hospital. The majority of the cases with P. multocida possessed some underlying pulmonary diseases (seven cases, 70%), inactive lung tuberculosis or bronchiectasis. There were compromised hosts such as high ages person, steroids dependent person and diabetes mellitus patients. P. multocida was almost susceptible to antibioticus (penicillin and cephalosporins), although some erythromycin resistant strains were identified. The cats' oral cavities in our two cases were cultured and P. multocida were isolated. In our survey the prevalence of this organism is as high as 85% in cats. Our data suggests that patients who are in the high infection risk category are easily infected to P. multocida.

Adolescent↗

Partial characterization of R-plasmids from Pasteurella multocida isolated from turkeys.

Pasteurella multocida, isolated from turkeys during an outbreak of septicemic disease ("fowl cholera"), was found to be resistant to tetracycline, streptomycin, and sulfonamides. Agarose gel electrophoretic analysis of DNA from these isolates indicated the presence of extrachromosomal elements. Plasmid DNA was isolated by cesium chloride-ethidium bromide density centrifugation. Escherichia coli was transformed to antimicrobic resistance with this DNA. Two plasmids were isolated. One of these plasmids had a buoyant density of 1.7158 g/cm(3) (56.9 mol% guanine plus cytosine) and a molecular weight of 4.4 x 10(6) and conferred resistance to tetracycline, streptomycin, and sulfonamides. The other, having a buoyant density of 1.7198 g/cm(3) (61 mol% guanine plus cytosine) and a molecular weight of 3.44 x 10(6), conferred resistance to streptomycin and sulfonamides. Streptomycin resistance was mediated by streptomycin phosphotransferase. Compatibility group testing indicated that neither plasmid belonged to any of 13 compatibility groups (of conjugal plasmids). Both plasmids were also found to be compatible with three small, nonconjugative resistance plasmids.

Animals↗

Antibiotic susceptibilities of human isolates of Pasteurella multocida.

Seventeen human strains of Pasteurella multocida, biochemically similar to, if not identical with, isolates of animal origins, were tested for susceptibility to 16 antimicrobial agents utilizing a microtiter broth dilution technique. Ten of these isolates were also tested against 11 antibiotics by disk diffusion. The most active drugs with respect to the median minimal inhibitory concentration (micrograms per milliliter) were tetracycline (0.09), penicillin G (0.78), ampicillin (0.78), carbenicillin (1.56), cephalothin (1.56), and chloramphenicol (1.56). With the exception of tetracycline, the median minimal inhibitory concentration and minimal bactericidal concentration values were equal or differed by no more than a factor of two. The semisynthetic penicillins clindamycin, erythromycin, and aminoglycosides had relatively low activities, suggesting that these agents would be poor choices for the treatment of P. multocida infections.

Anti-Bacterial Agents↗

Lack of in vitro efficacy of oral forms of certain cephalosporins, erythromycin, and oxacillin against Pasteurella multocida.

The in vitro susceptibility of human isolates of Pasteurella multocida to oral antimicrobial agents from our current study and from a review of the literature suggests that dicloxacillin (oxacillin), erythromycin, clindamycin, cephalexin, cefaclor, and cefadroxil should not be used for empiric therapy of animal bite wounds. Agents that were consistently active against P. multocida were penicillin, ampicillin, amoxicillin-clavulanic acid, tetracycline, minocycline, chloramphenicol, trimethoprim-sulfamethoxazole, and cefuroxime. Possible reasons for the confusion regarding the activity of oral cephalosporins are addressed.

Animals↗

Resistance to antimicrobial agents and prevalence of R plasmids in Pasteurella multocida from turkeys.

One hundred fifty-three isolates of Pasteurella multocida, representing the causative agent of 95% of all known outbreaks of fowl cholera occurring in California meat and breeder turkeys from August 1985 through February 1987, were examined for susceptibility to antimicrobial agents. Of the 153 isolates, 6 were shown to be resistant to one or more antimicrobial agents. Of the six resistant isolates, five contained R plasmids. All but one of the R plasmids were small (6 to 7 megadaltons) and nonconjugative, encoding resistance to tetracycline or kanamycin, streptomycin, and sulfonamides; the other was large (70 megadaltons) and conjugative, transferring resistance to kanamycin, streptomycin, sulfonamides, and tetracycline to P. multocida and Escherichia coli. The three plasmids encoding resistance to tetracycline alone appeared identical.

Animals↗

Evidence of a dormant but infective state of the fish pathogen Pasteurella piscicida in seawater and sediment.

The stability of Pasteurella piscicida strains in seawater and sediment microcosms at different temperatures (6 and 20 degrees C) was investigated during a 1-month period. Three strains of P. piscicida showed similar survival kinetics. By a standard plate count method they survived in water and sediment for only 6 to 12 days, depending on the strain and type of microcosm. During this starvation period, the metabolic activity of the cells was reduced by more than 80%. Culturable cells of each P. piscicida strain persisted better in sediment than in water, as well as at 20 degrees C compared to 6 degrees C. However, in all the microcosms, the acridine orange direct counts remained at about 10(5) cells per ml during the experimental period, which demonstrated that P. piscicida possesses a capacity to enter a viable but not culturable state. Moreover, dormant cells were always resuscitated by the addition of fresh medium to the microcosms, since we recovered numbers of culturable cells similar to the acridine orange direct counts. These resuscitated cells exhibited the same respiration rate as that seen prior to the start of the experiments. Although the biochemical, physiological, and serological characteristics; lipopolysaccharides; membrane proteins; and plasmid content of P. piscicida strains were unaffected during the starvation conditions, the dormant cells were smaller (dwarf cells) and had increased surface hydrophobicity. The starved cells maintained their infectivity and pathogenic potential for fish, with 50% lethal doses similar to those of the original strains.

Animals↗

Iron uptake by Pasteurella piscicida and its role in pathogenicity for fish.

We evaluated the iron uptake mechanisms in Pasteurella piscicida strains as well as the effect of iron overload on the virulence of these strains for fish. With this aim, the capacity of the strains to obtain iron from transferrin and heme compounds as well as their ability to overcome the inhibitory activity of fish serum was analyzed. All the P. piscicida strains grew in the presence of the iron chelator ethylene-diamine-di (O-hydroxyphenyl acetic acid) or of human transferrin, which was used by a siderophore-mediated mechanism. The chemical tests and cross-feeding assays showed that P. piscicida produced a siderophore which was neither a phenolate nor a hydroxamate. Cross-feeding assays as well as preliminary chromatographic analysis suggest that this siderophore may be chemically related to multocidin. All the P. piscicida isolates utilized hemin and hemoglobin as an iron source, since the virulence of the strains increased when the fish were preinoculated with these compounds. This effect was stronger in the avirulent strains (50% lethal dose was reduced by 4 logs when fish were pretreated with hemin or hemoglobin). Only the pathogenic P. piscicida isolates were resistant to the bactericidal action of the fresh fish serum. The nonpathogenic strains grew in fish serum only when it was heat-inactivated or when it was supplemented with ferric ammonium citrate, hemin, or hemoglobin. In all the strains, at least three iron-regulated outer membrane proteins (IROMPs) (105, 118, and 145 kDa) were increased when the strains were cultured in iron-restricted medium.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Vibrio sp. strain NM 10, isolated from the intestine of a Japanese coastal fish, has an inhibitory effect against Pasteurella piscicida.

Vibrio sp. strain NM 10 with an inhibitory activity against Pasteurella piscicida K-III was isolated from the intestine of a spotnape ponyfish (Leiognathus nuchalis). This bacterium efficiently produced an antibacterial substance after growth at 20 degrees C for 24 h on 1/5 PYBG agar prepared with 50% seawater at pHs of 7.5 to 9.0. The antibacterial substance was heat labile and proteinaceous, with a molecular mass of less than 5 kDa, possibly a bacteriocin or a bacteriocin-like substance.

Animals↗

Isolation and sequencing of a temperate transducing phage for Pasteurella multocida.

A temperate bacteriophage (F108) has been isolated through mitomycin C induction of a Pasteurella multocida serogroup A strain. F108 has a typical morphology of the family Myoviridae, presenting a hexagonal head and a long contractile tail. F108 is able to infect all P. multocida serogroup A strains tested but not those belonging to other serotypes. Bacteriophage F108, the first P. multocida phage sequenced so far, presents a 30,505-bp double-stranded DNA genome with cohesive ends (CTTCCTCCCC cos site). The F108 genome shows the highest homology with those of Haemophilus influenzae HP1 and HP2 phages. Furthermore, an F108 prophage attachment site in the P. multocida chromosome has been established to be inside a gene encoding tRNA(Leu). By using several chromosomal markers that are spread along the P. multocida chromosome, it has been demonstrated that F108 is able to perform generalized transduction. This fact, together with the absence of pathogenic genes in the F108 genome, makes this bacteriophage a valuable tool for P. multocida genetic manipulation.

Animals↗

Immunization of mice with components of Pasteurella multocida.

Log-phase cells of Pasteurella multocida strain P-1059 were used to prepare isolated culture filtrate, cell wall, and cytoplasmic components. Culture filtrate was further separated by column chromatography. A portion of cytoplasm and culture filtrate was conjugated to ferritin by means of metaxylylene diisocyanate. Cell walls induced more protection in mice than the conjugated or unconjugated cytoplasm or culture filtrate. The cell walls caused edema and erythema when given intradermally in rabbits, whereas cytoplasm and culture filtrate produced dermal necrosis. The first of four chromatographically separated fractions of culture filtrate was possibly more immunogenic in mice than cell walls. This fraction was less reactive intradermally in rabbits than cell walls but more reactive than the other fractions.

Acrylates↗

Serological response to Pasteurella multocida NanH sialidase in persistently colonized rabbits.

Pasteurella multocida is a mucosal pathogen that colonizes the upper respiratory system of rabbits. Respiratory infections can result, but the bacteria can also invade the circulatory system, producing abscesses or septicemia. P. multocida produces extracellular sialidase activity, which is believed to augment colonization of the respiratory tract and the production of lesions in an active infection. Previously, it was demonstrated that some isolates of P. multocida contain two unique sialidase genes, nanH and nanB, that encode enzymes with different substrate specificities (S. Mizan, A. D. Henk, A. Stallings, M. Meier, J. J. Maurer, and M. D. Lee, J. Bacteriol. 182:6874-6883, 2000). We developed a recombinant antigen enzyme-linked immunosorbent assay (ELISA) based on the NanH sialidase of P. multocida and demonstrated that rabbits that were experimentally colonized with P. multocida produce detectable anti-NanH immunoglobulin M (IgM) and IgG in serum, although they demonstrated no clinical signs of pasteurellosis. In addition, clinically ill pet rabbits infected with P. multocida possessed IgM and/or IgG antibody against NanH. The NanH ELISA may be useful for the diagnosis of P. multocida infections in sick rabbits as well as for screening for carriers in research rabbit colonies.

Animals↗

Effect of ovalbumin aerosol exposure on colonization of the porcine upper airway by Pasteurella multocida and effect of colonization on subsequent immune function.

Seventy-three piglets were weaned at 1 week of age, randomly assigned to 10 groups (A to J), accommodated in stainless steel exposure chambers, and exposed continuously to a controlled environment containing aerosolized ovalbumin. The concentrations of ovalbumin dust were as follows (milligrams per cubic meter): A and F, 16.6; B and G, 8.4; C and H, 4.2; D and I, 2.1; E and J, 0. At weekly intervals, the pigs were bled via venipuncture and anesthetized for nasal lavage and tonsilar biopsies performed for subsequent bacteriologic analysis. At 2 weeks of age, the pigs in groups A to E were challenged with toxigenic Pasteurella multocida (10(8) CFU pig(-1)), and at 6 weeks of age, the pigs were euthanatized. At postmortem, the extent of turbinate atrophy was assessed on the snout sections by using a morphometric index. Exposure to aerial ovalbumin resulted in a dose-dependent increase in serum antiovalbumin immunoglobulin G (IgG; P < 0.001) and serum antiovalbumin IgA (P < 0.001). Exposure also caused a significant increase in the numbers of P. multocida organisms isolated from the upper respiratory tract (P < 0.001) and a corresponding increase in turbinate atrophy, as judged by the morphometric index (P < 0.001). Concurrent challenge with P. multocida and ovalbumin resulted in a significant decrease in both the IgG and IgA responses to ovalbumin (P < 0.001). These results show that ovalbumin exposure increases pig susceptibility to P. multocida colonization and that toxigenic P. multocida modifies the serum IgG and IgA responses to ovalbumin in the pig. Both of these effects may enhance the virulence of this respiratory pathogen and so influence the pathogenesis of atrophic rhinitis in pigs.

Aerosols↗

Immune mechanism in Pasteurella multocida-infected mice.

Various immunizing procedures were tested for their effect on the growth of Pasteurella multocida in specific pathogen-free CD-1 mice. Two injections with killed vaccine incorporated into Freund complete adjuvant gave excellent protection against parenteral challenge, but less to control the growth of an aerogenic inoculum. Protection with adjuvant-treated preparation was always superior to that seen with non-adjuvant-treated vaccines. A killed saline-suspended vaccine introduced aerogenically marginally increased resistance to small (2 to 5 mean lethal doses) aerogenic challenges. Live mycobacterium bovis (BCG) vaccine immunopotentiated a single dose of heat-killed P. multocida vaccine in terms of the protective immune response against a subsequent footpad challenge. Hyperimmune mouse serum, administered intraperitoneally, intramuscularly, or intravenously 1 to 7 days prior to subcutaneous challenge with 500 to 5,000 P. multocida, was highly protective. Mice injected in the thigh with 0.2 ml of hyperimmune serum prior to footpad challenge frequently developed a severe inflammatory response with local swelling within 48 h; these lesions often became severly abscessed with time. The passively protected, footpad-infected mice developed active immunity against a subsequent challenge given into the opposite footpad.

Administration, Intranasal↗