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Current Microbiology Apr 2002Capsular serotype A strains of Pasteurella multocida of avian origin express a 40-kDa lipoprotein (Plp-40) thought to attach the extracellular polysaccharide to the cell... (Comparative Study)
Comparative Study
Capsular serotype A strains of Pasteurella multocida of avian origin express a 40-kDa lipoprotein (Plp-40) thought to attach the extracellular polysaccharide to the cell surface. The objective of the present study was to assess the prevalence of Plp-40 in P. multocida strains of disparate serotypes and host origins, as well as other pathogenic members of the family Pasteurellaceae. Exponential-phase reference and clinical isolates were radiolabeled with [3H]-palmitate, lysed to obtain whole-cell protein fractions, and analyzed using SDS-PAGE and fluorography to assess lipoprotein content. The ability to produce Plp-40 was found to be conserved among certain P. multocida reference and clinical strains of different host origins including avian, human, porcine, bovine, feline, canine, ovine, and cervine, but not rabbit. Production of a 40-kDa lipoprotein was exhibited by all clinical isolates of Pasteurella aerogenes, Pasteurella pneumotropica, Actinobacillus suis, Actinobacillus suis-like organism, and Actinobacillus pleuropneumoniae examined, but not Pasteurella (Mannheimia) haemolytica, Actinobacillus lignieresii, or Haemophilus spp. These data suggest that, while not all Pasteurellaceae are able to produce a 40-kDa lipoprotein under the present experimental conditions, expression is somewhat conserved among diverse isolates of disparate host origins.
Topics: Animals; Bacterial Capsules; Birds; Cats; Cattle; Chickens; Deer; Dogs; Electrophoresis, Polyacrylamide Gel; Ferrets; Humans; Lipoproteins; Molecular Weight; Pasteurella Infections; Pasteurella multocida; Pasteurellaceae; Rabbits; Rats; Serotyping; Sheep; Swine; Turkeys
PubMed: 11910502
DOI: 10.1007/s00284-001-0047-2 -
Molecular Phylogenetics and Evolution Mar 2010A phylogenomic approach was used to generate an amino acid phylogeny for 12 whole genomes representing 10 species in the family Pasteurellaceae. Orthology of genes was...
A phylogenomic approach was used to generate an amino acid phylogeny for 12 whole genomes representing 10 species in the family Pasteurellaceae. Orthology of genes was determined using an approach similar to OrthologID (http://nypg.bio.nyu.edu/orthologid/about.html) and resulted in the generation of a matrix with 3130 genes with 1,194,615 aligned amino acid characters of which 239,504 characters are phylogenetically informative. Phylogenetic analysis of the concatenated matrix using all standard approaches (maximum parsimony, maximum likelihood, and Bayesian analysis) results in a single extremely robust phylogenetic hypothesis for the species examined in this study. Remarkably, no single gene partition gives the same tree as the concatenated analysis. By analyzing partitioned support in the data matrix, we show that there is very little negative support emanating from individual gene partitions to suggest that the concatenated hypothesis is not tenable. The large number of characters in the matrix allows us to test hypotheses concerning missing data and character number in phylogenomic studies, and we conclude that matrices constructed using genome level information are very robust to missing data. We show that a very large number of concatenated gene sequences (>160) are needed to reliably obtain the same topology as the overall analysis.
Topics: Bayes Theorem; DNA, Bacterial; Genes, Bacterial; Genome, Bacterial; Genomics; Likelihood Functions; Models, Genetic; Pasteurellaceae; Phylogeny; Sequence Analysis, DNA
PubMed: 19686857
DOI: 10.1016/j.ympev.2009.08.010 -
Journal of Veterinary Diagnostic... May 2011A set of polymerase chain reaction (PCR) assays for identification of the most important Pasteurellaceae species encountered in cats and dogs were developed. Primers for...
A set of polymerase chain reaction (PCR) assays for identification of the most important Pasteurellaceae species encountered in cats and dogs were developed. Primers for Pasteurella multocida were designed to detect a fragment of the kmt, a gene encoding the outer-membrane protein. Primers specific to Pasteurella canis, Pasteurella dagmatis, and Pasteurella stomatis were based on the manganese-dependent superoxide dismutase gene (sodA) and those specific to [Haemophilus] haemoglobinophilus on species-specific sequences of the 16S ribosomal RNA gene. All the primers were tested on respective reference and control strains and applied to the identification of 47 canine and feline field isolates of Pasteurellaceae. The PCR assays were shown to be species specific, providing a valuable supplement to phenotypic identification of species within this group of bacteria.
Topics: Animals; Cat Diseases; Cats; Dog Diseases; Dogs; Pasteurella; Pasteurella Infections; Pasteurellaceae; Pasteurellaceae Infections; Phenotype; Poland; Polymerase Chain Reaction
PubMed: 21908285
DOI: 10.1177/1040638711403434 -
Journal of Microbiological Methods Nov 2013The rodents Pasteurellaceae have to be excluded from the specified pathogen free experimental animal facilities. Despite the biological and economic importance of...
The rodents Pasteurellaceae have to be excluded from the specified pathogen free experimental animal facilities. Despite the biological and economic importance of Pasteurellaceae in relation to experimental animals just a few molecular based methods are available for their detection and identification. The aim of the present investigation was to develop a multiplex PCR assay allowing detection of all rodent Pasteurellaceae and identification of [Pasteurella] pneumotropica biotype Jawetz, [P.] pneumotropica biotype Heyl and [Actinobacillus] muris, as the most prevalent members of the group. For this, a Pasteurellaceae common forward primer located on the 16S rRNA gene was used in conjunction with four different reverse primers specific for [P.] pneumotropica biotype Jawetz, [P.] pneumotropica biotype Heyl, [A.] muris and a common reverse primer for all rodent Pasteurellaceae, all targeting the 16S-23S rRNA internal transcribed spacer sequences. The performance characteristics of the assay were tested against 125 Pasteurellaceae isolates belonging to eleven different species and including 34 strains of [P.] pneumotropica biotype Jawetz, 44 strains of [P.] pneumotropica biotype Heyl and 37 strains of [A.] muris. Additionally, eight other mouse associated bacterial species which could pose a diagnostic problem were included. The assay showed 100% sensitivity and specificity. Identification of the clinical isolates was validated by ITS profiling and when necessary by 16S rRNA gene sequencing. This multiplex PCR represents the first molecular tool able to detect and differentiate in a single assay among the Pasteurellaceae found in laboratory mouse and may become a reliable alternative to the present diagnostic methods.
Topics: Animals; Base Sequence; DNA Primers; DNA, Bacterial; DNA, Ribosomal Spacer; Mice; Molecular Sequence Data; Multiplex Polymerase Chain Reaction; Pasteurellaceae; Phenotype; RNA, Ribosomal, 16S; RNA, Ribosomal, 23S; Rodentia; Sensitivity and Specificity; Sequence Analysis, DNA; Species Specificity
PubMed: 24055385
DOI: 10.1016/j.mimet.2013.09.005 -
Veterinary Microbiology Sep 1993NAD dependent members of the family Pasteurellaceae were cultured from the nasal cavity, surface and cut surface of the tonsils, and from the apical and caudal lobes of...
NAD dependent members of the family Pasteurellaceae were cultured from the nasal cavity, surface and cut surface of the tonsils, and from the apical and caudal lobes of the lungs of 303 slaughterhouse pigs from 5 different herds in order to obtain information on the ecology of these bacteria. The specimens were plated on two different selective agar media using a special dilution technique that resulted in a good separation of individual colonies. Bacteriological results were compared with serological and pathological findings. The bacteriological examination demonstrated that NAD dependent Pasteurellaceae belonging to the taxa previously described could be isolated from the surface and cut surface of the tonsils, and from lungs with and without gross pathologic lesions. Haemophilus parasuis was detected mainly from the nasal cavity, and Actinobacillus pleuropneumoniae mainly from the surface and cut surface of the tonsils (42%). From two herds, 19% and 24% respectively of the animals without antibodies against A. pleuropneumoniae serotypes 1 and 2 harboured the bacteria mainly in the tonsils. This may reflect a very recent infection or may suggest that A. pleuropneumoniae can colonize the tonsils without inducing a serologic reaction. Serological and bacteriological evidence of more than one serotype in the same herd indicates that natural infection with one serotype does not necessarily protect against another.
Topics: Actinobacillus pleuropneumoniae; Animals; Antibodies, Bacterial; Culture Media; Female; Lung; Male; NAD; Palatine Tonsil; Pasteurellaceae; Pasteurellaceae Infections; Respiratory System; Respiratory Tract Infections; Swine; Swine Diseases
PubMed: 8273273
DOI: 10.1016/0378-1135(93)90093-m -
Laboratory Animals Oct 2009Monitoring of rodents for Pasteurellaceae infection may be carried out by the polymerase chain reaction (PCR). We tested which of 17 rodent Pasteurellaceae strains were... (Comparative Study)
Comparative Study
Monitoring of rodents for Pasteurellaceae infection may be carried out by the polymerase chain reaction (PCR). We tested which of 17 rodent Pasteurellaceae strains were detected by three PCR primer sets. By phylogenetic analysis, 12 strains were assigned to the Rodent cluster and five strains to other clusters, namely the Somnus cluster, Pasteurella sensu stricto, Actinobacillus sensu stricto, the Mannheimia and Rossii cluster. A primer set developed to detect biotype Heyl [Pasteurella] pneumotropica produced amplicons from three strains and appeared specific for this taxon. A primer set developed to detect biotype Jawetz [P.] pneumotropica produced amplicons from the [P.] pneumotropica type strain and two other strains within the Rodent cluster. A primer set as described by Bootz and his co-workers (Bootz F, Kirschnek S, Nicklas W, Wyss SK, Homberger FR. Detection of Pasteurellaceae in rodents by polymerase chain reaction analysis. Lab Anim Sci 1998;48:542-6) for the detection of all Pasteurellaceae indeed detected all bacterial strains examined. Bootz's primer set should be used to monitor rodents for Pasteurellaceae infection by PCR as FELASA recommends the monitoring of rodents for all Pasteurellaceae taxa. Health monitoring reports should specify the primer set(s) used for PCR testing rodents for Pasteurellaceae infection.
Topics: Animals; Cricetinae; DNA Primers; DNA, Bacterial; Environmental Monitoring; Guinea Pigs; Laboratory Animal Science; Mice; Murinae; Pasteurellaceae; Pasteurellaceae Infections; Phylogeny; Polymerase Chain Reaction; Rats; Rodentia
PubMed: 19505934
DOI: 10.1258/la.2009.0070131 -
Canadian Journal of Veterinary Research... Jan 2001Ninety pharyngeal tonsils were collected from 2-year-old American bison (Bison bison) bulls and sampled for members of the Pasteurellaceae family. Particular attention...
Ninety pharyngeal tonsils were collected from 2-year-old American bison (Bison bison) bulls and sampled for members of the Pasteurellaceae family. Particular attention was paid to seasonal incidence and antimicrobial resistance in serotypes and biovariants. Multiple strains of Pasteurella haemolytica (39%), P. trehalosi (68%), P. multocida (34%) and Haemophilus somnus (13%) were cultured from 86 out of the 90 (96%) tonsil samples. Pasteurella trehalosi was the most common and evenly distributed of the organisms recovered. Pasteurella haemolytica was found in fewer numbers than P. trehalosi, but showed an increase in number of isolates recovered with each sampling period. Pasteurella multocida, both A and D capsular types, was recovered from all sampling periods. No serotype pattern was observed in any of the animal groups sampled. One hundred twenty-seven of 147 (86%) of the isolates were resistant to at least 1 antibiotic, 95/147 (65%) to at least 2 different antibiotics, and 16/147 (11%) to at least 3 antibiotics. The most common resistance pattern observed was to neomycin and spectinomycin (73/147) (49%).
Topics: Animals; Bison; Carrier State; Drug Resistance, Microbial; Microbial Sensitivity Tests; Neomycin; Palatine Tonsil; Pasteurellaceae; Pasteurellaceae Infections; Seasons; Serotyping; Spectinomycin
PubMed: 11227200
DOI: No ID Found -
New plasmid tools for genetic analysis of Actinobacillus pleuropneumoniae and other pasteurellaceae.Applied and Environmental Microbiology Oct 2009We have generated a set of plasmids, based on the mobilizable shuttle vector pMIDG100, which can be used as tools for genetic manipulation of Actinobacillus...
We have generated a set of plasmids, based on the mobilizable shuttle vector pMIDG100, which can be used as tools for genetic manipulation of Actinobacillus pleuropneumoniae and other members of the Pasteurellaceae. A tandem reporter plasmid, pMC-Tandem, carrying promoterless xylE and gfpmut3 genes downstream of a multiple-cloning site (MCS), can be used for identification of transcriptional regulators and conditions which favor gene expression from different cloned promoters. The ability to detect transcriptional regulators using the tandem reporter system was validated in A. pleuropneumoniae using the cloned rpoE (sigma(E)) promoter (P). The resulting plasmid, pMCrpoEP, was used to identify a mutant defective in production of RseA, the negative regulator of sigma(E), among a bank of random transposon mutants, as well as to detect induction of sigma(E) following exposure of A. pleuropneumoniae to ethanol or heat shock. pMCsodCP, carrying the cloned sodC promoter of A. pleuropneumoniae, was functional in A. pleuropneumoniae, Haemophilus influenzae, Haemophilus parasuis, Mannheimia haemolytica, and Pasteurella multocida. Two general expression vectors, pMK-Express and pMC-Express, which differ in their antibiotic resistance markers (kanamycin and chloramphenicol, respectively), were constructed for the Pasteurellaceae. Both plasmids have the A. pleuropneumoniae sodC promoter upstream of the gfpmut3 gene and an extended MCS. Replacement of gfpmut3 with a gene of interest allows complementation and heterologous gene expression, as evidenced by expression of the Haemophilus ducreyi nadV gene in A. pleuropneumoniae, rendering the latter NAD independent.
Topics: Base Sequence; Cloning, Molecular; DNA, Bacterial; Genes, Bacterial; Genes, Reporter; Genetic Vectors; Molecular Biology; Molecular Sequence Data; Pasteurellaceae; Plasmids; Promoter Regions, Genetic; Sequence Analysis, DNA; Transcription, Genetic
PubMed: 19666733
DOI: 10.1128/AEM.00809-09 -
Proceedings of the National Academy of... Mar 2004The DNA-uptake signal sequence (USS) of the bacterium Haemophilus influenzae is highly over-represented in its genome (1,471 copies of the core sequence AAGTGCGGT), and...
The DNA-uptake signal sequence (USS) of the bacterium Haemophilus influenzae is highly over-represented in its genome (1,471 copies of the core sequence AAGTGCGGT), and DNA fragments containing USS are preferentially taken up by competent cells. Because this bias favors uptake of conspecific DNA, USSs are often considered a kind of mate recognition system in bacteria, acting as species-specific barriers against uptake of unrelated DNA. However, the H. influenzae USS is highly over-represented in the genomes of three otherwise-divergent Pasteurellaceae species (Pasteurella multocida, Haemophilus somnus, and Actinobacillus actinomycetemcomitans, 927, 1,205, and 1,760 copies, respectively), suggesting that USSs do not always limit exchange. USSs in all these genomes are mainly in coding regions and show no orientation bias around the chromosome, weakening proposed USS functions in transcription termination and chromosome replication. Alignment of homologous genes was used to determine evolutionary relationships between individual USSs. Most H. influenzae USSs were found to have perfect or imperfect homologs (USS at the same location) in at least one other species, and most USSs in the other species had perfect or imperfect homologs in H. influenzae. These homologies suggest that the use of a common USS is due to inheritance of the USS-based uptake system from a common ancestor of the Pasteurellaceae, and it indicates that individual USSs can be evolutionarily stable elements of their genomes. The pattern is consistent with a molecular drive model of USS evolution, with new USSs arising by mutation and preferentially spread to new genomes by the biased DNA-uptake system.
Topics: Base Pairing; Base Sequence; Biological Transport; DNA, Bacterial; Evolution, Molecular; Haemophilus; Pasteurellaceae; Sequence Alignment; Sequence Homology, Nucleic Acid
PubMed: 15070749
DOI: 10.1073/pnas.0306366101 -
Laboratory Animals Jul 1995Thirty Pasteurellaceae strains isolated from gerbil, guineapig, hamster, mouse, muskrat and rat were reinvestigated and reclassified after comparison with reference...
Thirty Pasteurellaceae strains isolated from gerbil, guineapig, hamster, mouse, muskrat and rat were reinvestigated and reclassified after comparison with reference strains. Strains originally described as Pasteurella pneumotropica were reclassified as [Pasteurella] pneumotropica Heyl biotype (7), [P.] pneumotropica Jawetz biotype (1), Pasteurella dagmatis (1) or Taxon 22 (2). Strains previously reported as Actinobacillus sp. were reclassified as [P.] pneumotropica biotype Jawetz (3), P. dagmatis (3) or Taxon 6 (7). Strains earlier described as Pasteurella gallinarum were renamed as SP group pasteurella (4) or Taxon 25 (2). Some of these reclassified Pasteurellaceae have not been reported previously in rodents. The present findings underline the importance of extended characterization of isolates and comparison with references strains to avoid misclassification within the family Pasteurellaceae Pohl 1981.
Topics: Actinobacillus; Animals; Bacterial Typing Techniques; Cricetinae; Guinea Pigs; Mice; Pasteurella; Pasteurellaceae; Phenotype; Rats; Rodentia; Serotyping
PubMed: 7564217
DOI: 10.1258/002367795781088342