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PubMed2006

Antimicrobial susceptibilities of Pasteurella strains isolated from humans.

The in vitro susceptibilities of 192 consecutive clinical strains of Pasteurella spp. isolated between 1996 and 2003 from soft tissue pus (n = 146), respiratory tract specimens (n = 38) and blood (n = 8) were studied by an agar dilution method. All isolates were susceptible to minocycline, cefotaxime, ofloxacin, ciprofloxacin and levofloxacin. Most strains were susceptible to moxifloxacin, amoxicillin, azithromycin and clarithromycin, whereas lower susceptibility rates to telithromycin (89.4%) were observed among respiratory tract isolates.

Anti-Bacterial Agents↗
PubMed2005

Pasteurella multocida meningitis: case report and review of the literature.

Pasteurella multocida forms part of the normal flora in the nasopharynx of many domestic and wild animals. Most human P. multocida infections are soft tissue infections due to animal bites. P. multocida meningitis is a rare condition. We report a case of P. multocida meningitis with a complicated outcome and review the literature of this condition.

Aged↗
PubMed2006

How does Pasteurella multocida respond to the host environment?

Pasteurella multocida is a Gram-negative bacterial pathogen, which causes diseases of economic importance in a wide range of animal species. The response of P. multocida to the host environment has been analysed at the transcription level, using DNA microarrays, and at the protein-expression level, using proteomics techniques. Furthermore, a growing number of P. multocida-directed mutants have been assessed for their ability to cause disease. Although technical impediments mean that it is currently difficult to analyse bacterial responses at the earliest stages of infection, it is clear that during later stages of infection the bacteria encounter host niches that require them to modify the expression of genes involved in central energy metabolism and in the uptake of various nutrients such as iron and amino acids. Furthermore, in vitro experiments have defined the varying bacterial responses to low iron and to different iron sources, including haemoglobin and transferrin. To date, most P. multocida genes shown to be upregulated during infection are involved in nutrient acquisition and metabolic processes, indicating that true virulence genes might be constitutively expressed, upregulated only during initial stages of infection or upregulated at levels below current detection limits.

Adaptation, Physiological↗
PubMed2004

Characterization of two lipoproteins in Pasteurella multocida.

An in vivo expression technology (IVET) system was previously developed and used to identify Pasteurella multocida genes, which are upregulated during infection of the host. Of the many genes identified, two encoded products which showed similarity to the Haemophilus influenzae lipoproteins, protein D and PCP, which have been shown to stimulate heterologous immunity against infection with H. influenzae. Therefore, the lipoprotein homologues in P. multocida, designated GlpQ and PCP, were investigated. GlpQ and PCP were shown to be lipoproteins by demonstrating that post-translational processing of the proteins was inhibited by globomycin. The P. multocida GlpQ homologue showed glycerophosphodiester phosphodiesterase enzyme activity, indicating that it is a functional homologue of other characterized GlpQ enzymes. Using surface immunoprecipitation, PCP was found to be surface exposed, but GlpQ was not. Non-lipidated forms of GlpQ and PCP were expressed and purified from Escherichia coli and used to vaccinate mice. However, mice were not protected from challenge with live P. multocida. The lipoproteins were then expressed in E. coli in the lipidated form and used to vaccinate mice and chickens. Protection against challenge with live P. multocida was not observed.

Animals↗
PubMed2004

Genomic-scale analysis of Pasteurella multocida gene expression during growth within liver tissue of chickens with fowl cholera.

We have recently reported the gene expression profile of Pasteurella multocida during growth in the blood of chickens with fowl cholera. Here we report the gene expression profile of P. multocida during growth in the livers of similarly infected chickens. We compared expression profiles of bacteria harvested from the livers of infected chickens with late-stage fowl cholera with those of bacteria grown in rich medium. Independent analysis of bacterial expression profiles from three individual chickens indicated that 93 P. multocida genes were always differentially expressed during growth in liver tissue. Of these 93 genes, 49 were upregulated and 44 downregulated in the host. Many of the upregulated genes were involved in energy production and conversion (9/49) and carbohydrate transport and metabolism (8/49), and a number of these have been shown to be induced under anaerobic conditions in other species. The downregulated genes were generally of unknown or poorly characterised functions (14/44). Comparison of the differentially regulated gene sets identified for growth in liver with those identified previously for growth in blood allowed the identification of a core set of 13 upregulated and 16 downregulated genes that were differentially regulated in at least five of the six infections studied.

Anaerobiosis↗
PubMedDate not supplied

Construction and virulence of a Pasteurella multocida fhaB2 mutant in turkeys.

Pasteurella multocida is the causative agent of fowl cholera. The organism can occur as a commensally in the naso-pharyngeal region of apparently healthy animals and it can be a primary or secondary pathogen in the disease process of birds. The complete genome of an avian strain of P. multocida has been sequenced and was shown to possess two filamentous hemagglutinin genes designated fhaB1 and fhaB2. Filamentous hemagglutinin transposon mutants of a bovine strain of P. multocida are attenuated in mice. Here, we report the construction of an fhaB2 P. multocida mutant in an avian strain P-1059 (A:3). The fhaB2 mutant and the parent were assessed for virulence in turkeys by intranasal and intravenous challenge. Inactivation of fhaB2 resulted in a high degree of attenuation when turkeys were challenged intranasally and to a lesser degree when intravenously administered. Resistance of the fhaB2 mutant and parent strain to killing by serum complement was similar.

Animals↗
PubMed2006

[Pasteurella dagmatis spondylodiscitis in a diabetic patient].

INTRODUCTION: Healthy pets contact are able to induce unusual and severe diseases. CLINICAL CASE: This case reports the medical history of a dorsal spondylodiscitis in a diabetic patient admitted for a chronic wound of his toes. He had a long-standing history of regular consumption of alcohol and tobacco. Blood bottles and biopsy of intervertebral disc infected discovered Pasteurella dagmatis, commonly colonizing the oropharynx of healthy dogs and cats. In this case, licking of his injured toe by his dog was the likely source of entry of the organism. DISCUSSION: We found no identical cases in the medical literature. Diabetes mellitus and other immunocompromised disorders justify to change behaviours toward domestic animals.

Animals↗
PubMed2006

Characterization of avian strains of Pasteurella multocida by restriction endonuclease and amplified fragment length polymorphism.

Avian strains of Pasteurella multocida were typed by employing restriction endonuclease analysis (REA) and single enzyme-amplified fragment length polymorphism (AFLP) to evaluate their applicability for epidemiological studies of fowl cholera outbreaks. A total of 72 strains isolated from different avian species (chicken, duck, turkey, quail and goose) belonging to various geographical regions of India were characterized. REA using two different enzymes HhaI and HpaII produced 9 and 18 clusters respectively, whereas Single enzyme-AFLP recognized 32 patterns out of 72 strains typed. The study indicated that REA using HpaII is a simple and resource efficient method, however, further typing with more stringent and rapid method like Single enzyme-AFLP, could drastically enhance investigation in epidemiological studies of fowl cholera outbreaks.

Animals↗
PubMed2005

Synergistic action of E. coli endotoxin and Pasteurella multocida type A for the induction of bronchopneumonia in pigs.

This study aimed to investigate whether Escherichia coli endotoxin (LPS) may predispose the lung to an infection with Pasteurella multocida type A (Pma) and to determine the LPS concentration needed to reproduce clinical signs of bronchopneumonia. Twenty-four hours before inoculating Pma or sterile growth medium, piglets were tracheally instilled with 10, 100 or 400 microg/kg LPS. Cough, body temperature, daily weight gain (DWG) bronchoalveolar lavage fluid (BALF) cells and volume of pneumonic lung were measured. Changes in breathing pattern (Penh) were assessed by whole body barometric plethysmography. No significant changes were observed in Pma-treated or in control animals. Each LPS doses induced DWG reduction while the higher generated a severe subacute interstitial pneumonia causing hyperthermia and an increase in Penh. The combination of the lower LPS doses with Pma produced an asymptomatic bronchopneumonia leading to DWG reduction, rise in Penh and an increase in BALF macrophages and neutrophils. With 400 microg/kg LPS, Pma worsened the inflammatory process as illustrated by cough, hyperthermia, major DWG reduction and by a greater Penh response. Lung lesions consisted of severe exudative bronchopneumonia. We concluded that LPS may negatively influence growth, predispose to persisting lung inflammatory process and promote Pma infection depending on the dose previously administered.

Animals↗
PubMed2005

Identification of avian strains of Pasteurella multocida in India by conventional and PCR assays.

The prevalence of capsular and somatic serotypes were studied among 123 Pasteurella multocida strains isolated from chickens (n=94), ducks (22), quails (4), turkeys (2) and geese (1) from different geographical regions of India. All strains exhibited similar cultural and morphological characteristics. Ninety-two of the isolates belonged to serotype A:1, the most prevalent serotype, with serotypes A:3, A:1,3, D:3 and F:3 having two isolates each. Only one isolate was positive for serotypes A:4 and D:1. Twenty isolates were untyped. A multiplex capsular PCR assay generated amplicons of sizes approximately 460, approximately 1044, approximately 657 and approximately 854 bp in 106 isolates identified as capsular serotype-A, 15 in serotype D and two in serotype F. Capsular types B and E were not detected in any of the avian isolates studied. The present findings suggest that a multiplex capsular PCR assay may be suitable for the rapid initial identification serotypes P. multocida during epidemiological studies of fowl cholera.

Animals↗
PubMed2003

A 39 kDa protein mediates adhesion of avian Pasteurella multocida to chicken embryo fibroblast cells.

To clarify the role of avian Pasteurella multocida capsule in pathogenesis, adhesion of capsulated strains P-1059, X-73 and Pm-18, and noncapsulated strains P-1059B, Pm-1 and Pm-3 to chicken embryo fibroblast (CEF) cells was compared. Number of adherent organisms of the capsulated strains to CEF cells were approximately three times as much as noncapsulated strains indicating that adhesive properties were enhanced by the presence of bacterial capsule. Pretreatments of the bacterial cells with heat, trypsin, or with antiserum caused a marked decrease in adhesion of capsulated strain P-1059 and its noncapsulated variant P-1059B. However, depolymerization of capsular hyaluronic acid with high dose of hyaluronidase enhanced adhesion of these strains. Combined treatments of the bacterial cells with both hyaluronidase and trypsin significantly (P < 0.05) inhibited the adherence of strain P-1059 as compared to the treatment only with trypsin, but strain P-1059B was not affected. SDS-PAGE profiles of crude capsular extract (CCE) prepared from capsulated strain P-1059 and its noncapsulated variant P-1059B grown on dextrose starch agar (DSA) plates by heating at 56 degrees C in a 2.5% NaCl solution demonstrated eight protein bands of 28, 34, 36, 39, 52, 56, 63 and 93 kDa. The 28, 34 and 36 kDa proteins were commonly major for both strains, and the 39 kDa protein was major only for strain P-1059 but poor in strain P-1059B. Outer membrane protein (OMP) profiles were identical with a major protein at 34 kDa and four minor proteins between the two strains. The adhesion of strain P-1059 and strain P-1059B to CEF cells was inhibited significantly (P < 0.01) by treatment with rabbit antisera against P-1059, P-1059B, CCE or 39 kDa protein of strain P-1059 as compared to the treatment with either PBS or with normal rabbit serum. These results indicated that an antigenic 39 kDa protein in the capsule may be responsible for adhesion of avian P. multocida type A strains to CEF cells as a virulence factor.

Animals↗
PubMed2003

Capsule thickness and amounts of a 39 kDa capsular protein of avian Pasteurella multocida type A strains correlate with their pathogenicity for chickens.

Capsule thickness of avian Pasteurella multocida type A strains was determined by transmission electron microscopy after labeling with polycationic ferritin and compared with their pathogenicity for chickens. The capsule thickness of P. multocida strains Pm-18 and X-73 was 81.4 and 50.1 nm on average, respectively. These strains were highly virulent for chicken, whereas the less virulent strains Pm-1 and Pm-3 had a thin and irregular capsule, 21.0 and 29.8 nm on average, respectively. However, the thickest capsule was observed in strain P-1059, 101.2 nm on average, and the strain revealed moderate virulence. The noncapsulated variant P-1059B, which was derived from strain P-1059, revealed low virulence. The six P. multocida strains were examined with regard to protein content on the capsule of organisms. Amounts of total proteins of crude capsular extract (CCE) from capsulated strains were approximately twice those of the noncapsulated strains. The amount of an antigenic 39 kDa protein in the CCE were found to correlate with the capsule thickness, since heavily capsulated strains exhibited the greatest amount, whereas noncapsulated strains including noncapsulated and low virulent variant P-1059B possessed little 39 kDa protein. The results demonstrated that the capsule thickness and the quantity of a 39 kDa capsular protein of avian P. multocida type A strains correlated with their pathogenicity for chickens.

Animals↗
PubMed2004

Pasteurella multocida contains multiple immunogenic haemin- and haemoglobin-binding proteins.

Iron-dependent outer membrane proteins (IROMPs) play an important role in bacterial pathogenesis and present several attributes of potential vaccine candidates. TBLASTN analysis of the Pasteurella multocida Pm70 genome using the same molecules of other bacterial pathogens as a query identified eight putative haemin and haemoglobin receptors for this organism. Quantitative binding assays have demonstrated that the proteins PM0040, PM0236, PM0741, PM1081, PM1428, PM0592 and HgbA bind both haemin and haemoglobin, whereas PM0576 and PM1282 ORFs only bind either haemoglobin or haemin, respectively. Furthermore, Western blot analysis showed that P. multocida-infected mice generate specific antibodies against PM0040, PM0236, PM0741, PM1081, PM1428, PM0592 and HgbA proteins. Nevertheless, inoculation of mice with any single one of these receptors alone did not protect against P. multocida infection.

Animals↗
PubMed2004

Characterisation of bovine strains of Pasteurella multocida and comparison with isolates of avian, ovine and porcine origin.

One hundred and fifty-three bovine Pasteurella multocida strains recovered primarily from cases of pneumonia and mastitis in England and Wales over an 11-year period were characterised by capsular PCR typing, comparison of outer membrane protein (OMP) profiles, and multilocus sequence analysis. All of the strains were of capsular type A with the exception of a single capsular type F isolate. Thirteen distinct OMP profiles (OMP-types) were identified based mainly on molecular mass heterogeneity of the heat-modifiable (OmpA) and porin (OmpH) proteins. However, 85% of the isolates were represented by just five OMP-types and 39% of the strains were of a single OMP-type. Multilocus sequence analysis revealed a limited degree of genetic diversity among bovine P. multocida isolates; strains of the same OMP-type have identical genetic backgrounds and represent distinct clones. Analysis of OMP variation was more discriminating than multilocus sequence analysis because strains of different OMP-types had the same, or similar, genetic backgrounds. The association of a small number of clones with the majority of cases of bovine pneumonia suggests that these clones have an increased capacity to cause disease compared to less frequently recovered clones. Molecular mass heterogeneity of OmpA and OmpH, in strains of the same or similar genetic background, suggests that these proteins are subject to diversifying selection within the host and might play important roles in host-pathogen interactions. Comparison of the OMP profiles of bovine isolates with those of avian, ovine and porcine strains showed that a high proportion of the respiratory tract infections in each of these species are caused by different strains of P. multocida. However, the presence of small numbers of closely related strains in more than one host species suggests that transmission of bacteria between different host species is also a factor in the population biology of P. multocida.

Animals↗
PubMed2004

Characterization of a 39kDa capsular protein of avian Pasteurella multocida using monoclonal antibodies.

The role of a 39kDa protein of avian Pasteurella multocida in pathogenesis of fowl cholera was investigated using monoclonal antibodies (Mabs). Mabs were prepared by immunization of BALB/c mice with a crude capsular extract (CCE) of P. multocida strain P-1059 (serovar A:3). Totally eight hybridomas producing Mab were obtained. Immunoblot analysis of the hybridomas revealed that all the Mabs recognized a 39kDa protein of CCE. Treatment of CCE antigen with proteinase K or periodic acid indicated that the epitope recognized was proteinaceous. The Mabs reacted with a major 39kDa protein of CCE from encapsulated strains but not with any protein of non-capsulated strains indicating that a direct correlation between encapsulation and the 39kDa protein. Immunoelectron microscopy on strain P-1059 and the non-capsulated derivative P-1059B (serovar -:3) reacting with the Mabs and gold-labeled anti-mouse IgG indicated that the protein is associated with the capsule. The Mabs significantly inhibited the adherence of encapsulated P. multocida strains to chicken embryo fibroblast cells, but only slightly that of non-capsulated strains. Mice passively immunized with the Mabs were protected from lethal challenge with virulent strains P-1059 and X-73 (serovar A:1). Thus the capsular 39kDa protein was determined to be an adherence factor and a cross-protective antigen of avian P. multocida type A strains.

Animals↗
PubMed2004

Differentiation of Pasteurella multocida isolates from cases of atrophic rhinitis in pigs from Zimbabwe by RAPD and ribotyping.

Atrophic rhinitis in pigs is rarely reported in Southern Africa. To determine the relationship between Pasteurella multocida clones from clinical cases of atrophic rhinitis, twenty-one strains were characterised by selected phenotypic and genotypic methods. Biochemical analysis classified 18 strains as P. multocida subspecies multocida, whilst the remainder were grouped into separate unassigned biotypes. Capsular groups A (16/21) and D (l/21) were found among the isolates by PCR. Four ribotype patterns were obtained following HpaII ribotyping, whilst random amplification of polymorphic DNA (RAPD) revealed three main clusters. However, subclusters were also noted for each RAPD cluster. Our results indicate that RAPD offers a better discrimination of strains than ribotyping and that none of the phenotypic characters were directly related to the genotypic clusters.

Animals↗
PubMed2004

Invasion of chicken embryo fibroblast cells by avian Pasteurella multocida.

Invasion of chicken embryo fibroblast (CEF) cells by the virulent encapsulated Pasteurella multocida strains P-1059 (serovar A:3) and X-73 (serovar A:1) and an avirulent noncapsulated derivative P-1059B (serovar -:3) was investigated. The number of intracellular bacteria increased for all the strains after 2, 4 and 6 h post-inoculation to CEF cells. By 6 h post-inoculation, the number of invaded bacteria of encapsulated strains was significantly higher than noncapsulated strain and reached 150- and 112-fold for strains P-1059 and X-73, respectively, while it was 9-fold for strain P-1059B as compared to the number of invaded bacteria recovered after 2 h post-inoculation. Electron microscopy of invasion by encapsulated strains showed that the bacteria were adhering to CEF cells membrane after 1 h of inoculation. By 4-h, one or two bacteria were detected within membrane-bound vacuoles of the intracellular space. The number of intracellular bacteria markedly increased at 14 h post-inoculation. Invasion of all strains was inhibited significantly when the monolayers were treated with periodic acid (P<0.001) or trypsin (P<0.05). The treatment of bacteria with hyaluronidase did not affect invasion. The present results indicate that avian P. multocida capsular type A strains are invasive and that the receptor on CEF cell surface might be glycoprotein.

Animals↗