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Journal of Bacteriology Nov 2013Pasteurella multocida is a Gram-negative multispecies pathogen and the causative agent of fowl cholera, a serious disease of poultry which can present in both acute and...
Pasteurella multocida Heddleston serovar 3 and 4 strains share a common lipopolysaccharide biosynthesis locus but display both inter- and intrastrain lipopolysaccharide heterogeneity.
Pasteurella multocida is a Gram-negative multispecies pathogen and the causative agent of fowl cholera, a serious disease of poultry which can present in both acute and chronic forms. The major outer membrane component lipopolysaccharide (LPS) is both an important virulence factor and a major immunogen. Our previous studies determined the LPS structures expressed by different P. multocida strains and revealed that a number of strains belonging to different serovars contain the same LPS biosynthesis locus but express different LPS structures due to mutations within glycosyltransferase genes. In this study, we report the full LPS structure of the serovar 4 type strain, P1662, and reveal that it shares the same LPS outer core biosynthesis locus, L3, with the serovar 3 strains P1059 and Pm70. Using directed mutagenesis, the role of each glycosyltransferase gene in LPS outer core assembly was determined. LPS structural analysis of 23 Australian field isolates that contain the L3 locus revealed that at least six different LPS outer core structures can be produced as a result of mutations within the LPS glycosyltransferase genes. Moreover, some field isolates produce multiple but related LPS glycoforms simultaneously, and three LPS outer core structures are remarkably similar to the globo series of vertebrate glycosphingolipids. Our in-depth analysis showing the genetics and full range of P. multocida lipopolysaccharide structures will facilitate the improvement of typing systems and the prediction of the protective efficacy of vaccines.
Topics: Amino Acid Sequence; Gene Expression Regulation, Bacterial; Genetic Variation; Lipopolysaccharides; Molecular Sequence Data; Pasteurella multocida
PubMed: 23974032
DOI: 10.1128/JB.00779-13 -
PloS One 2015[Pasteurella] pneumotropica biotypes Jawetz and Heyl and [Actinobacillus] muris are the most prevalent Pasteurellaceae species isolated from laboratory mouse. However,...
[Pasteurella] pneumotropica biotypes Jawetz and Heyl and [Actinobacillus] muris are the most prevalent Pasteurellaceae species isolated from laboratory mouse. However, mechanisms contributing to their high prevalence such as the ability to form biofilms have not been studied yet. In the present investigation we analyze if these bacterial species can produce biofilms in vitro and investigate whether proteins, extracellular DNA and polysaccharides are involved in the biofilm formation and structure by inhibition and dispersal assays using proteinase K, DNase I and sodium periodate. Finally, the capacity of the biofilms to confer resistance to antibiotics is examined. We demonstrate that both [P.] pneumotropica biotypes but not [A.] muris are able to form robust biofilms in vitro, a phenotype which is widely spread among the field isolates. The biofilm inhibition and dispersal assays by proteinase and DNase lead to a strong inhibition in biofilm formation when added at the initiation of the biofilm formation and dispersed pre-formed [P.] pneumotropica biofilms, revealing thus that proteins and extracellular DNA are essential in biofilm formation and structure. Sodium periodate inhibited the bacterial growth when added at the beginning of the biofilm formation assay, making difficult the assessment of the role of β-1,6-linked polysaccharides in the biofilm formation, and had a biofilm stimulating effect when added on pre-established mature biofilms of [P.] pneumotropica biotype Heyl and a majority of [P.] pneumotropica biotype Jawetz strains, suggesting that the presence of β-1,6-linked polysaccharides on the bacterial surface might attenuate the biofilm production. Conversely, no effect or a decrease in the biofilm quantity was observed by biofilm dispersal using sodium periodate on further biotype Jawetz isolates, suggesting that polysaccharides might be incorporated in the biofilm structure. We additionally show that [P.] pneumotropica cells enclosed in biofilms were less sensitive to treatment with amoxicillin and enrofloxacin than planktonic bacteria. Taken together, these findings provide a first step in understanding of the biofilm mechanisms in [P.] pneumotropica, which might contribute to elucidation of colonization and pathogenesis mechanisms for these obligate inhabitants of the mouse mucosa.
Topics: Animals; Anti-Bacterial Agents; Biofilms; Deoxyribonuclease I; Endopeptidase K; Mice; Microbial Sensitivity Tests; Microscopy, Confocal; Pasteurella pneumotropica; Periodic Acid
PubMed: 26430880
DOI: 10.1371/journal.pone.0138778 -
Antimicrobial Agents and Chemotherapy Jun 2017Animal bite wounds affect more than 5 million Americans annually, resulting in 300,000 emergency department visits, 10,000 hospitalizations, and an untold number of...
Activity of Pexiganan and 10 Comparator Antimicrobials against 234 Isolates, Including 93 Pasteurella Species and 50 Anaerobic Bacterial Isolates Recovered from Animal Bite Wounds.
Animal bite wounds affect more than 5 million Americans annually, resulting in 300,000 emergency department visits, 10,000 hospitalizations, and an untold number of physician office visits. Various forms of topical therapy are empirically self-employed by many patients prior to seeking medical attention. Pexiganan, a 22-amino-acid synthetic cationic analogue of the peptide magainin II, acts by selectively damaging bacterial cell membranes. We determined the MICs for pexiganan and other antimicrobial agents often used for treatment of bite wounds. Most isolates were from U.S. patients, and ∼10% were from European and Canadian patients. The comparator antimicrobials studied were penicillin, amoxicillin-clavulanate, piperacillin-tazobactam, meropenem, clindamycin, doxycycline, moxifloxacin, ceftriaxone, linezolid, and metronidazole. The MICs of pexiganan were 32 μg/ml (against subsp. ), 16 μg/ml ( subsp. , , and ), 8 μg/ml (), 8 μg/ml (), 2 μg/ml (, , and group), 16 μg/ml (), 64 μg/ml (), 4 μg/ml (), 32 μg/ml (), and 64 μg/ml (). The concentration of pexiganan in the cream used was 8,000 μg/ml, more than 60 to 100 times the highest MIC obtained. Pexiganan exhibited a broad range of antimicrobial activity, showing potential for treating animal bite infections. A clinical trial seems warranted.
Topics: Amoxicillin-Potassium Clavulanate Combination; Animals; Anti-Bacterial Agents; Anti-Infective Agents; Antimicrobial Cationic Peptides; Bacteria, Anaerobic; Bites and Stings; Clindamycin; Doxycycline; Fluoroquinolones; Linezolid; Meropenem; Metronidazole; Microbial Sensitivity Tests; Moxifloxacin; Pasteurella; Penicillanic Acid; Penicillins; Piperacillin; Piperacillin, Tazobactam Drug Combination; Thienamycins
PubMed: 28373186
DOI: 10.1128/AAC.00246-17 -
International Journal of Infectious... May 2004Two episodes of peritoneal dialysis-associated peritonitis, which occurred four months apart and were both due to Pasteurella multocida, were noted in a 73 year old... (Review)
Review
OBJECTIVES
Two episodes of peritoneal dialysis-associated peritonitis, which occurred four months apart and were both due to Pasteurella multocida, were noted in a 73 year old woman. This report aims to describe the clinical history of these episodes and the microbiological investigations that were undertaken. The relevant literature will also be discussed.
METHODS AND RESULTS
Basic microbiological tests identified the organism as Pasteurella multocida, and further work at a specialist laboratory classified it as Pasteurella multocida subsp. multocida. Pulsed field gel electrophoresis confirmed that the strains isolated from the two clinical episodes originated from the same clone. A literature search was performed, looking particularly for patients who experienced more than one episode of peritonitis caused by Pasteurella spp, whether due to recurrence or re-infection.
CONCLUSIONS
It is likely that the infection resulted from a domestic cat, as there was evidence of a cat bite to the dialysis tubing in the period between the two episodes. Re-infection with two identical strains of pasteurella is more probable than relapse, for reasons discussed. Strict hygiene and avoiding contact between dialysis tubing and domestic animals must be emphasised to try to prevent pasteurella and other animal-associated infections in this already vulnerable population.
Topics: Aged; Animals; Cats; Female; Humans; Kidney Failure, Chronic; Pasteurella Infections; Pasteurella multocida; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis
PubMed: 15109592
DOI: 10.1016/j.ijid.2003.10.004 -
Journal of Clinical Microbiology Mar 1981Fifty serotyped isolates each of Pasteurella multocida and Pasteurella haemolytica were tested on the API 20E strip (Analytab Products, Plainview, N.Y.), the Oxi/Ferm...
Fifty serotyped isolates each of Pasteurella multocida and Pasteurella haemolytica were tested on the API 20E strip (Analytab Products, Plainview, N.Y.), the Oxi/Ferm tube (Roche Diagnostics, Nutley, N.J.), and the Minitek system (BBL Microbiology Systems, Cockeysville, Md.). None of the rapid test systems reliable identified these organisms. With the API system, discrepancies between expected and actual results for the oxidase test and nitrate test frequently resulted in misidentification or no identification. The Minitek system misidentified 68% of the P. haemolytica isolates. The Minitek identification of Pasteurella depends on 100% positive xylose reactions, whereas only 56% of the P. haemolytica strains were positive for xylose fermentation. The Oxy/Ferm system, instead of giving a definitive identification, in most instances merely placed Pasteurella in a category of similar organisms.
Topics: Bacteriological Techniques; Evaluation Studies as Topic; Fermentation; Oxidation-Reduction; Pasteurella; Reagent Kits, Diagnostic; Serotyping
PubMed: 7016894
DOI: 10.1128/jcm.13.3.433-437.1981 -
Applied and Environmental Microbiology Sep 2005Pasteurella multocida is a highly infectious, facultative intracellular bacterium which causes fowl cholera in birds. This study reports, for the first time, the...
Pasteurella multocida is a highly infectious, facultative intracellular bacterium which causes fowl cholera in birds. This study reports, for the first time, the observed interaction between P. multocida and free-living amoebae. Amoebal trophozoites were coinfected with fowl-cholera-causing P. multocida strain X-73 that expressed the green fluorescent protein (GFP). Using confocal fluorescence microscopy, GFP expressing X-73 was located within the trophozoite. Transmission electron microscopy of coinfection preparations revealed clusters of intact X-73 cells in membrane-bound vacuoles within the trophozoite cytoplasm. A coinfection assay employing gentamicin to kill extracellular bacteria was used to assess the survival and replication of P. multocida within amoebae. In the presence of amoebae, the number of recoverable intracellular X-73 cells increased over a 24-h period; in contrast, X-73 cultured alone in assay medium showed a consistent decline in growth. Cytotoxicity assays and microscopy showed that X-73 was able to lyse and exit the amoebal cells approximately 18 h after coinfection. The observed interaction between P. multocida and amoebae can be considered as an infective process as the bacterium was able to invade, survive, replicate, and lyse the amoebal host. This raises the possibility that similar interactions occur in vivo between P. multocida and host cells. Free-living amoebae are ubiquitous within water and soil environments, and P. multocida has been observed to survive within these same ecosystems. Thus, our findings suggest that the interaction between P. multocida and amoebae may occur within the natural environment.
Topics: Acanthamoeba; Animals; Green Fluorescent Proteins; Lobosea; Microscopy, Electron, Transmission; Pasteurella multocida; Plasmids
PubMed: 16151138
DOI: 10.1128/AEM.71.9.5458-5464.2005 -
BMC Veterinary Research Apr 2019Pasteurella multocida is responsible for significant economic losses in pigs worldwide. In clinically diseased pigs, most P. multocida isolates are characterised as...
BACKGROUND
Pasteurella multocida is responsible for significant economic losses in pigs worldwide. In clinically diseased pigs, most P. multocida isolates are characterised as subspecies multocida, biovar 2 or 3 and capsular type A or D; however, there is little information regarding subspecies, biovars, and other capsular types of P. multocida isolates in Korea. Here, we provided information covering an extended time period regarding P. multocida in pigs with pneumonia in Korea using phenotypic and genotypic characterisations and data associated with the minimum inhibitory concentrations.
RESULTS
The overall prevalence of P. multocida between 2008 and 2016 was 16.8% (240/1430), with 85% of the P. multocida isolates (204/240) coinfected with other respiratory pathogens. Of the 240 isolates, 166 were included in this study; all of these P. multocida isolates were characterised as subspecies multocida and the most prevalent phenotypes were represented by biovar 3 (68.7%; n = 114) and capsular type A (69.9%; n = 116). Additionally, three capsular type F isolates were identified, with this representing the first report of such isolates in Korea. All biovar 1 and 2 isolates were capsular types F and A, respectively. The virulence-associated gene distribution was variable; all capsular type A and D isolates harboured pmHAS and hsf-1, respectively (P < 0.001), with type F (biovar 1) significantly correlated with hsf-1 (P < 0.05) and pfhA (P < 0.01), biovar 2 highly associated with pfhA and pmHAS, and biovar 3 significantly correlated with hsf-1, pmHAS, and hgbB (P < 0.001), whereas biovar 13 was related only to hgbB (P < 0.05). The highest resistance rate was found to be to oxytetracycline (63.3%), followed by florfenicol (16.3%).
CONCLUSIONS
P. multocida subspecies multocida, biovar 3, and capsular type A was the most prevalent isolate in this study, and our findings indicated the emergence of capsular type F in Korea. Moreover, prudent use of oxytetracycline and florfenicol is required because of the identified high resistance rates. Further studies are required for continuous monitoring of the antimicrobial resistance, prevalence, and epidemiological characterisation of P. multocida, and experimental infection models are needed to define the pathogenicity of capsular type F.
Topics: Animals; Bacterial Proteins; Gene Expression Regulation, Bacterial; Pasteurella Infections; Pasteurella multocida; Pneumonia, Bacterial; Republic of Korea; Swine; Swine Diseases
PubMed: 31023320
DOI: 10.1186/s12917-019-1861-5 -
Veterinary Research Mar 2021Fowl cholera caused by Pasteurella multocida exerts a massive economic burden on the poultry industry. Lipopolysaccharide (LPS) is essential for the growth of P....
Fowl cholera caused by Pasteurella multocida exerts a massive economic burden on the poultry industry. Lipopolysaccharide (LPS) is essential for the growth of P. multocida genotype L1 strains in chickens and specific truncations to the full length LPS structure can attenuate bacterial virulence. Here we further dissected the roles of the outer core transferase genes pcgD and hptE in bacterial resistance to duck serum, outer membrane permeability and virulence in ducks. Two P. multocida mutants, ΔpcgD and ΔhptE, were constructed, and silver staining confirmed that they all produced truncated LPS profiles. Inactivation of pcgD or hptE did not affect bacterial susceptibility to duck serum and outer membrane permeability but resulted in attenuated virulence in ducks to some extent. After high-dose inoculation, ΔpcgD showed remarkably reduced colonization levels in the blood and spleen but not in the lung and liver and caused decreased injuries in the spleen and liver compared with the wild-type strain. In contrast, the ΔhptE loads declined only in the blood, and ΔhptE infection caused decreased splenic lesions but also induced severe hepatic lesions. Furthermore, compared with the wild-type strain, ΔpcgD was significantly attenuated upon oral or intramuscular challenge, whereas ΔhptE exhibited reduced virulence only upon oral infection. Therefore, the pcgD deletion caused greater virulence attenuation in ducks, indicating the critical role of pcgD in P. multocida infection establishment and survival.
Topics: Animals; Bacterial Proteins; Ducks; Lipopolysaccharides; Pasteurella Infections; Pasteurella multocida; Poultry Diseases; Transferases
PubMed: 33663572
DOI: 10.1186/s13567-021-00910-4 -
Brazilian Journal of Microbiology :... Dec 2022Swine pasteurellosis is one of the most economically important diseases of pig caused by Pasteurella multocida (P. multocida) capsular types A and D. These organisms are...
Swine pasteurellosis is one of the most economically important diseases of pig caused by Pasteurella multocida (P. multocida) capsular types A and D. These organisms are commensals and opportunistic pathogens in the upper respiratory tract in pig. In the present study, we extracted whole outer membrane proteins (OMP) from P. multocida capsular types A and D and were mixed together in the ratio of 1:1 forming bivalent outer-membrane proteins. The bivalent OMP was adsorbed onto aluminum hydroxide nanoparticles. The size of aluminum hydroxide nanoparticles adsorbed outer membrane protein was found to be in the range of 125 to 130 nm. We observed that aluminum hydroxide nanoparticles adjuvanted bivalent OMP-based vaccine elicited quicker immune kinetics in terms of IgG response as compared to aluminum hydroxide microparticles adjuvanted bivalent bacterin vaccine against P. multocida capsular type A and D.
Topics: Swine; Animals; Pasteurella multocida; Aluminum Hydroxide; Vaccines, Combined; Bacterial Outer Membrane Proteins; Pasteurella Infections; Nanoparticles
PubMed: 35922692
DOI: 10.1007/s42770-022-00795-1 -
European Journal of Medical Research Dec 2011Pasteurella species, widely known as indigenous organisms in the oral and gastrointestinal floras of many wild and domestic animals, are important pathogens in both...
A case of wound dual infection with Pasteurella dagmatis and Pasteurella canis resulting from a dog bite -- limitations of Vitek-2 system in exact identification of Pasteurella species.
BACKGROUND
Pasteurella species, widely known as indigenous organisms in the oral and gastrointestinal floras of many wild and domestic animals, are important pathogens in both animals and humans. Human infections due to Pasteurella species are in most cases associated with infected injuries following animal bites. We encountered a rare case of dual infections caused by different two Pasteurella species occurred in a previously healthy 25-year-old female sustaining injury by a dog-bite.
METHODOLOGY
Exudates from the open wound of her dog-bite site, together with the saliva of the dog were submitted for bacteriological examination. Predominantly appearing grayish-white smooth colonies with almost the same colonial properties but slightly different glistening grown on chocolate and sheep blood agar plates were characterized morphologically by Gram's stain, biochemically by automated instrument using Vitek 2 system using GN cards together with commercially available kit system, ID-Test HN-20 rapid panels, and genetically by sequencing the 16S rRNA genes of the organism using a Taq DyeDeoxy Terminator Cycle Sequencing and a model 3100 DNA sequencer instrument.
RESULTS
The causative isolates from the dog-bite site were finally identified as P. canis and P. dagmatis from the findings of the morphological, cultural, and biochemical properties together with the comparative sequences of the 16S rRNA genes. Both the isolates were highly susceptible to many antibiotics and the patient was successfully treated with the administration of so-called the first generation cephalosporin, cefazolin followed by so-called the third generation cephalosporin, cefcapene pivoxil. The isolate from the dog was subsequently identified as P. canis, the same species as the isolate from the patient.
CONCLUSIONS
To the best of our knowledge, this was the second report of a dual infection with Pasteurella species consisting of P. dagmatis and P. canis resulting from a dog-bite, followed by the first report of dual infections due to P. dagmatis and P. multocida in 1988. Our isolate finally identified as P. dagmatis was misidentified as P. pneumotripica by means of the Vitek 2 system. The species name "P. dagmatis" was not included in the database of the system. It is also important for routine clinical microbiology laboratories to know the limitation of the automated Vitek 2 system for the accurate identification of Pasteurella species especially P. dagmatis. It should be emphasized that there still exists much room for improvement in Vitek 2 system. Significant improvement of Vitek 2 system especially in the identification of Pasteurella species is urgently desired.
Topics: Adult; Animals; Anti-Bacterial Agents; Bacterial Typing Techniques; Bites and Stings; Cefazolin; Cephalosporins; Dogs; Female; Humans; Pasteurella; Pasteurella Infections; Wound Infection
PubMed: 22112359
DOI: 10.1186/2047-783x-16-12-531