-
Journal of Zoo and Wildlife Medicine :... Dec 2017Eighty-two Pasteurellaceae isolates from marsupials characterized by phylogenetic analysis of rpoB gene sequences formed five distinct groups. Twenty-one strains from...
Eighty-two Pasteurellaceae isolates from marsupials characterized by phylogenetic analysis of rpoB gene sequences formed five distinct groups. Twenty-one strains from long-nosed potoroos ( Potorous tridactylus apicalis), spotted-tailed quolls ( Dasyurus maculatus), and eastern quolls ( Dasyurus viverrinus) made up group 1, which classified with Frederiksenia canicola. Group 2, 15 strains from Tasmanian devils ( Sarcophilus harrisii), common wombats ( Vombatus ursinus), common ring-tailed possums ( Pseudocheirus peregrinus), and eastern quolls, grouped with Pasteurella multocida. Three strains from koalas ( Phascolarctos cinereus) formed group 3 and clustered with Lonepinella koalarum. Group 4, 13 common wombat strains only distantly related to other Pasteurellaceae, probably represent a new genus. Finally, 29 strains from Tasmanian devils, spotted-tailed quolls and eastern quolls formed group 5 and clustered with 15 previously described Tasmanian devil strains, belonging to a yet unnamed Pasteurellaceae taxon. The results strongly indicate that Pasteurellaceae bacteria represent a part of the normal oral microbiota in marsupials.
Topics: Animals; Marsupialia; Mouth; Pasteurellaceae; Phylogeny
PubMed: 29297829
DOI: 10.1638/2017-0071.1 -
Methods in Molecular Biology (Clifton,... 2015Species of the family Pasteurellaceae play an important role as primary or opportunistic, predominantly respiratory, pathogens in domestic and wild animals. Some of them...
Species of the family Pasteurellaceae play an important role as primary or opportunistic, predominantly respiratory, pathogens in domestic and wild animals. Some of them cause severe disease with high economic losses in commercial animal husbandry. Hence, rapid and accurate differentiation of Pasteurellaceae is important and signifies a particular challenge to diagnostic laboratories. Identification and differentiation of Pasteurellaceae is mostly done using phenotypic tests or genetic identification based on sequence similarity of housekeeping genes, such as the rrs gene encoding the 16S ribosomal RNA (16S rRNA). Both approaches are time consuming, laborious, and costly, therefore often delaying the final diagnosis of disease or epidemics. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry represents an alternative rapid and reliable method for the differentiation of most members of the family Pasteurellaceae. It is able to differentiate within a few minutes the currently known 18 genera and most of the over 60 species and subspecies of Pasteurellaceae including many members encountered in veterinary diagnostic laboratories. A few closely related species and subspecies that cannot be discriminated by MALDI-TOF are easily identified further by complementary simple tests, such as hemolysis done simultaneously or routinely during pathogen isolation.
Topics: Animal Diseases; Animals; Databases, Factual; Pasteurellaceae; Pasteurellaceae Infections; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 25399101
DOI: 10.1007/978-1-4939-2004-4_18 -
International Journal of Systematic and... Aug 2018The aim of the investigation was to investigate the phylogeny of the 49 type strains of species of Pasteurellaceae and three genomospecies, which are available with...
The aim of the investigation was to investigate the phylogeny of the 49 type strains of species of Pasteurellaceae and three genomospecies, which are available with whole genomic sequences. The genomes were downloaded from National Center for Biotechnological Information and for three species of Avibacterium sequenced in the present investigation. From the predicted protein sequences of proteins, which were conserved in all genomes, 31 proteins were randomly selected for the study. The protein sequences were concatenated for each taxon, and a multiple alignment reconstructed for the 52 taxa. Phylogenetic analysis was performed by using the maximum-likelihood and neighbour-joining methods and confirmed the classification of the genera, which have been classified based on phylogenetic analysis of 16S rRNA gene sequences. The comparison linked [Haemophilus]parainfluezae and [Haemophilus] pittmania with Haemophilus influenzae (type species of genus) although at a much lower level than observed for Haemophilus aegyptius, H. influenzae and Haemophilus haemolyticus. The comparison documented that three, three and nine species of Actinobacillus, Pasteurella and Haemophilus, respectively, are not properly classified at genus level. Similar conclusions have been drawn by 16S rRNA gene sequence comparisons. The highest inter genus pairwise similarity was 88 % based on the comparison of the 31 concatenated protein sequences of the species included in the comparison. The level of intra genus pairwise similarity was also 88 %.
Topics: Actinobacillus; Amino Acid Sequence; Bacterial Typing Techniques; Base Composition; Base Sequence; DNA, Bacterial; Haemophilus; Pasteurella; Pasteurellaceae; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA
PubMed: 29923825
DOI: 10.1099/ijsem.0.002860 -
Journal of Applied Microbiology Apr 2010The aim of the present investigation was to identify and characterize Pasteurella-like isolates obtained from clinically affected psittacine birds.
AIMS
The aim of the present investigation was to identify and characterize Pasteurella-like isolates obtained from clinically affected psittacine birds.
METHODS AND RESULTS
A total of 37 isolates from psittacine birds tentatively classified with the family Pasteurellaceae were characterized phenotypically. The genetic relationship was investigated by sequencing of partial rpoB and 16S rRNA genes for selected isolates. The results obtained were compared with the data from 16 reference strains. Nine isolates were identified as Gallibacterium spp., 16 as Volucribacter spp. or Volucribacter-like, while 11 isolates were classified as taxon 44 of Bisgaard. A single isolate was identified as Pasteurella multocida.
CONCLUSIONS
Characterization of Pasteurellaceae by traditional methods is often inconclusive because of inconsistent reactions and phenotypic diversity. For the same reason, genotyping is essential to allow proper classification as demonstrated in the present study.
SIGNIFICANCE AND IMPACT OF THE STUDY
Limited information exists on the isolation and significance of Pasteurellaceae associated with clinically affected psittacine birds showing signs of digestive and/or respiratory disorders. The present investigations demonstrated that these organisms are widely distributed among clinically affected birds, but isolation of these taxa cannot be unambiguously correlated with the symptoms observed.
Topics: Animals; Bacterial Proteins; Bird Diseases; Molecular Sequence Data; Pasteurella Infections; Pasteurellaceae; Phenotype; Phylogeny; Psittaciformes; RNA, Ribosomal, 16S
PubMed: 19732214
DOI: 10.1111/j.1365-2672.2009.04518.x -
Journal of Microbiological Methods Apr 2012Species of the family Pasteurellaceae play an important role as primary or opportunistic animal pathogens. In veterinary diagnostic laboratories identification of this...
Species of the family Pasteurellaceae play an important role as primary or opportunistic animal pathogens. In veterinary diagnostic laboratories identification of this group of bacteria is mainly done by phenotypic assays while genetic identification based on housekeeping genes is mostly used for research and particularly important diagnostic samples. MALDI-TOF MS seems to represent a promising alternative to the currently practiced cumbersome, phenotypic diagnostics carried out in many veterinary diagnostic laboratories. We therefore assessed its application for animal associated members of the family Pasteurellaceae. The Bruker Biotyper 3.0 database was complemented with reference spectra of clinically relevant as well as commensal animal Pasteurellaceae species using generally five strains per species or subspecies and tested for its diagnostic potential with additional, well characterized field isolates. About 250 strains comprising 15 genera and more than 40 species and subspecies were included in the study, covering most representatives of the family. A high discrimination at the genus and species level was observed. Problematic discrimination was only observed with some closely related species and subspecies. MALDI-TOF MS was shown to represent a highly potent method for the diagnosis of this group of animal pathogens, combining speed, precision and low running costs.
Topics: Animals; Bacteriological Techniques; Pasteurellaceae; Pasteurellaceae Infections; Sensitivity and Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Veterinary Medicine
PubMed: 22343217
DOI: 10.1016/j.mimet.2012.02.001 -
Infection and Immunity Mar 2020Nasopharyngeal colonization with nontypeable (NTHi) is a prerequisite for developing NTHi-associated infections, including otitis media. Therapies that block NTHi...
Nasopharyngeal colonization with nontypeable (NTHi) is a prerequisite for developing NTHi-associated infections, including otitis media. Therapies that block NTHi colonization may prevent disease development. We previously demonstrated that , a closely related human commensal, can inhibit NTHi colonization and infection of human respiratory epithelium We have now assessed whether (a rodent commensal from the same family) can prevent NTHi colonization and disease using a murine NTHi otitis media model. Otitis media was modeled in BALB/c mice using coinfection with 1 × 10 PFU of influenza A virus MEM H3N2, followed by intranasal challenge with 5 × 10 CFU of NTHi R2866 Spec Mice were pretreated or not with an intranasal inoculation of 5 × 10 CFU 24 h before coinfection. NTHi and viable counts and inflammatory mediators (gamma interferon [IFN-γ], interleukin-1β [IL-1β], IL-6, keratinocyte chemoattractant [KC], and IL-10) were measured in nasal washes and middle ear tissue homogenate. pretreatment decreased the median colonization density of NTHi from 6 × 10 CFU/ml to 9 × 10 CFU/ml ( = 0.0004). Only 1/12 -pretreated mice developed otitis media on day 5 compared to 8/15 mice with no pretreatment (8% versus 53%, = 0.0192). Inflammation, clinical score, and weight loss were also lower in -pretreated mice. We have demonstrated that a single dose of a closely related commensal can delay onset of NTHi otitis media Human challenge studies investigating prevention of NTHi colonization are warranted to reduce the global burden of otitis media and other NTHi diseases.
Topics: Administration, Intranasal; Animals; Antibiosis; Carrier State; Colony Count, Microbial; Cytokines; Disease Models, Animal; Haemophilus Infections; Haemophilus influenzae; Influenza A Virus, H3N2 Subtype; Mice, Inbred BALB C; Nasal Mucosa; Nasopharynx; Otitis Media; Pasteurellaceae
PubMed: 31964748
DOI: 10.1128/IAI.00685-19 -
Current Topics in Microbiology and... 2016The pathogen surface is usually the first site of interaction with the host. Histophilus somni was earlier thought to only have an outer membrane on its surface. Now it... (Review)
Review
The pathogen surface is usually the first site of interaction with the host. Histophilus somni was earlier thought to only have an outer membrane on its surface. Now it is known that the surface is composed of many virulence factors, including outer membrane proteins, lipooligosaccharide or endotoxin, a fibrillar network, and an exopolysaccharide. Outer membrane blebs, endotoxin, the fibrillar network, and the exopolysaccharide are also shed from the surface. This review will focus on the surface proteins of this pathogen that may colonize the mucosal surface of ruminants as a commensal or may cause pneumonia, septicemia, myocarditis, thrombotic meningoencephalitis, arthritis, and/or abortion. The major outer membrane protein has been well studied. Since its size and epitopes vary from strain to strain, it may be useful for typing strains. Iron-regulated OMPs have also received much attention because of their role in iron uptake for in vivo growth of H. somni. Other OMPs may be protective, based on passive immunization with monospecific antibodies and active immunization experiments. The surface and shed fibrillar network has been shown to be an immunoglobulin-binding protein in that it binds bovine IgG2 by the Fc portion. Two repeat domains (DR1 and DR2) have cytotoxic Fic motifs. Vaccine studies with recombinant DR2 are promising. Studies of the bacterial genome as well as comparison of surface proteins of different strains from the various H. somni syndromes and carrier states will be discussed and have provided much insight into pathogenesis and protection.
Topics: Animals; Bacterial Outer Membrane Proteins; Cattle; Pasteurellaceae
PubMed: 26728061
DOI: 10.1007/82_2015_5011 -
PloS One 2008A novel fibronectin-binding protein from Pasteurella multocida (PM1665) that binds to the fibronectin type III(9-10) modules via two helix-hairpin-helix motifs has...
A novel fibronectin-binding protein from Pasteurella multocida (PM1665) that binds to the fibronectin type III(9-10) modules via two helix-hairpin-helix motifs has recently been described [1]. This protein shares homology with competence-related DNA-binding and uptake proteins (ComEA and ComE) from Gram-positive and Gram-negative bacteria. Here, we show that recombinant PM1665 (now designated ComE1) also binds to DNA through the same helix-hairpin-helix motifs required for fibronectin-binding. This binding to DNA is non sequence-specific and is confined to double-stranded DNA. We have cloned and expressed ComE1 proteins from five members of the Pasteurellaceae in order to further investigate the function(s) of these proteins. When expressed as recombinant GST-fusion proteins, all of the homologues bound both to fibronectin and to double-stranded DNA. Inactivation of the gene encoding the ComE1 homologue in Actinobacillus pleuropneumoniae indicates major roles for these proteins in at least two processes: natural transformation, and binding of bacteria to fibronectin.
Topics: Animals; Bacterial Proteins; DNA; DNA-Binding Proteins; Fibronectins; Humans; Pasteurellaceae; Recombinant Fusion Proteins; Surface Plasmon Resonance
PubMed: 19098981
DOI: 10.1371/journal.pone.0003991 -
BMC Biochemistry Nov 2011The Gram-negative bacterium Haemophilus influenzae is a glutathione auxotroph and acquires the redox-active tripeptide by import. The dedicated glutathione transporter...
BACKGROUND
The Gram-negative bacterium Haemophilus influenzae is a glutathione auxotroph and acquires the redox-active tripeptide by import. The dedicated glutathione transporter belongs to the ATP-binding cassette (ABC)-transporter superfamily and displays more than 60% overall sequence identity with the well-studied dipeptide (Dpp) permease of Escherichia coli. The solute binding protein (SBP) that mediates glutathione transport in H. influenzae is a lipoprotein termed GbpA and is 54% identical to E. coli DppA, a well-studied member of family 5 SBP's. The discovery linking GbpA to glutathione import came rather unexpectedly as this import-priming SBP was previously annotated as a heme-binding protein (HbpA), and was thought to mediate heme acquisition. Nonetheless, although many SBP's have been implicated in more than one function, a prominent physiological role for GbpA and its partner permease in heme acquisition appears to be very unlikely. Here, we sought to characterize five representative GbpA homologs in an effort to delineate the novel GbpA-family of glutathione-specific family 5 SBPs and to further clarify their functional role in terms of ligand preferences.
RESULTS
Lipoprotein and non-lipoprotein GbpA homologs were expressed in soluble form and substrate specificity was evaluated via a number of ligand binding assays. A physiologically insignificant affinity for hemin was observed for all five GbpA homologous test proteins. Three out of five test proteins were found to bind glutathione and some of its physiologically relevant derivatives with low- or submicromolar affinity. None of the tested SBP family 5 allocrites interacted with the remaining two GbpA test proteins. Structure-based sequence alignments and phylogenetic analysis show that the two binding-inert GbpA homologs clearly form a separate phylogenetic cluster. To elucidate a structure-function rationale for this phylogenetic differentiation, we determined the crystal structure of one of the GbpA family outliers from H. parasuis. Comparisons thereof with the previously determined structure of GbpA in complex with oxidized glutathione reveals the structural basis for the lack of allocrite binding capacity, thereby explaining the outlier behavior.
CONCLUSIONS
Taken together, our studies provide for the first time a collective functional look on a novel, Pasteurellaceae-specific, SBP subfamily of glutathione binding proteins, which we now term GbpA proteins. Our studies strongly implicate GbpA family SBPs in the priming step of ABC-transporter-mediated translocation of useful forms of glutathione across the inner membrane, and rule out a general role for GbpA proteins in heme acquisition.
Topics: Amino Acid Sequence; Bacteria; Bacterial Proteins; Glutathione; Membrane Transport Proteins; Molecular Sequence Data; Pasteurellaceae; Phylogeny; Sequence Alignment; Species Specificity
PubMed: 22087650
DOI: 10.1186/1471-2091-12-59 -
Current Topics in Microbiology and... 2016Histophilus somni is a commensal and an opportunistic bacterial pathogen associated with multisystemic diseases in cattle and sheep. Some strains of H. somni isolated... (Review)
Review
Histophilus somni is a commensal and an opportunistic bacterial pathogen associated with multisystemic diseases in cattle and sheep. Some strains of H. somni isolated from the genital tract of cattle are biochemically and serologically similar to the pathogenic strains, but are relatively innocuous. Several virulence factors/mechanisms have been identified in H. somni, of which the phase-variable lipooligosaccharide, induction of apoptosis of host cells, intraphagocytic survival, and immunoglobulin Fc-binding proteins have been well characterized. The genomes of H. somni pneumonia strain 2336 and preputial strain 129Pt have also been sequenced, and comparative analyses of these genomes have provided novel insights into the role of horizontal gene transfer in the evolution of the respective strains. Continued analyses of the genomes of H. somni strains and comparing them to the newly sequenced genomes of other bacteria facilitated the identification of a putative integrative and conjugative element (designated ICEHso2336) encoding tetracycline resistance. Comparative genomics also showed that the uptake signal sequence (5'-AAGTGCGGT) of Haemophilus influenzae is abundant in H. somni and provided a genetic basis for the recalcitrance of some strains of this species to natural transformation. The post-genomic era for H. somni offered an opportunity for the functional characterization of genes identified by computational methods. This opportunity has been realized to a great extent by transcriptomic studies that have identified several small noncoding RNAs and new genes. These new discoveries and developments are expected to stimulate further in-depth investigations of H. somni, especially from the systems biology viewpoint.
Topics: Base Sequence; Genomics; Molecular Sequence Data; Mutagenesis; Pasteurellaceae; Plasmids; Transcriptome
PubMed: 26728065
DOI: 10.1007/82_2015_5009