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RNA (New York, N.Y.) May 2018is a Gram-negative bacterium responsible for many important animal diseases. While a number of virulence factors have been identified, very little is known about how...
is a Gram-negative bacterium responsible for many important animal diseases. While a number of virulence factors have been identified, very little is known about how gene expression and protein production is regulated in this organism. Small RNA (sRNA) molecules are critical regulators that act by binding to specific mRNA targets, often in association with the RNA chaperone protein Hfq. In this study, transcriptomic analysis of the strain VP161 revealed a putative sRNA with high identity to GcvB from and serovar Typhimurium. High-throughput quantitative liquid proteomics was used to compare the proteomes of the VP161 wild-type strain, a mutant, and a GcvB overexpression strain. These analyses identified 46 proteins that displayed significant differential production after inactivation of , 36 of which showed increased production. Of the 36 proteins that were repressed by GcvB, 27 were predicted to be involved in amino acid biosynthesis or transport. Bioinformatic analyses of putative GcvB target mRNAs identified a strongly conserved 10 nucleotide consensus sequence, 5'-AACACAACAT-3', with the central eight nucleotides identical to the seed binding region present within GcvB mRNA targets in and Typhimurium. Using a defined set of seed region mutants, together with a two-plasmid reporter system that allowed for quantification of sRNA-mRNA interactions, this sequence was confirmed to be critical for the binding of the GcvB to the target mRNA, .
Topics: Amino Acids; Bacterial Proteins; Binding Sites; Escherichia coli; Gene Expression Profiling; High-Throughput Nucleotide Sequencing; Host Factor 1 Protein; Nucleotide Motifs; Pasteurella multocida; Protein Transport; RNA, Bacterial; RNA, Messenger; RNA, Small Untranslated; Regulon
PubMed: 29440476
DOI: 10.1261/rna.063248.117 -
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 -
BMC Microbiology Feb 2019This study provides biochemical and molecular genetic characteristics of P. multocida isolated from dead saigas in 1988, 2010-2015 on the territory of the Republic of...
BACKGROUND
This study provides biochemical and molecular genetic characteristics of P. multocida isolated from dead saigas in 1988, 2010-2015 on the territory of the Republic of Kazakhstan.
RESULTS
Bacteriological samples taken from carcasses of saiga antelope during mortality events recorded in West Kazakhstan in both 2010 and 2011 and in Kostanay in 2012 and 2015 confirmed the presence of P. multocida, according to morphological and biochemical characterisation. Only in the event of 2015 was the agent proven to be the causative agent of the disease observed, haemorrhagic septicaemia. In the other mortality events it is not certain if the organism was a primary aetiology or an incidental finding as confirmatory pathological investigation was not undertaken. The implemented phylogenetic analysis of ribosomal RNA 16S gene allowed us to identify Pasteurella strains isolated in 2010-2015 as P. multocida subspecies multocida. Capsular typing by PCR showed that the studied strains isolated from dead saiga in 2010, 2011, 2012 and 2015 belonged to serotype B. MLST analysis showed that these strains of P. multocida are of the capsule type B and form one clonal grouping with isolates ST64, ST44, ST45, ST46, ST44, ST47 which isolated from cases of hemorrhagic septicemia of animals in Hungary, Burma, Sri Lanka, Pakistan and Spain. Sixteen virulence genes of the five strains of P. multocida, isolated from saigas were studied using multiplex PCR. ptfA, ompA, ompH, oma87, plpB, fimA, hsf-2, pfhA, exbB, tonB, hgbA, fur, nanB, nanH and pmHAS genes were detected in all strains. The toxA gene was not identified in the studied strains. The phylogenies of these isolates is compared across saiga populations and years and the 2015 isolate was compared to that of an isolate from a disease outbreak in 1988 and the findings suggest that these isolated bacteria are stable commensals, opportunistically pathogenic, being phylogenetically uniform with very little genetic variation notable over the last 4 decades.
CONCLUSION
Isolation, phenotypic and genetic characterization of the P. multocida isolates inform understanding of the epidemiology of infection in saigas and predict virulent potential of these opportunistic bacteria.
Topics: Animals; Antelopes; Bacterial Typing Techniques; Genes, Bacterial; Hemorrhagic Septicemia; Kazakhstan; Multilocus Sequence Typing; Pasteurella Infections; Pasteurella multocida; Phylogeny; Serogroup; Virulence; Virulence Factors
PubMed: 30744550
DOI: 10.1186/s12866-019-1407-9 -
BMC Veterinary Research Jun 2020Pasteurella multocida is the etiological agent responsible for several diseases in a wide range of hosts around the world and thus, causes serious economic losses. Acute...
BACKGROUND
Pasteurella multocida is the etiological agent responsible for several diseases in a wide range of hosts around the world and thus, causes serious economic losses. Acute septicemia associated with capsular type B P. multocida has recently emerged in Europe and continuous outbreaks of these acute processes have been described in Spain since they were first detected in pigs in 2009 and cattle in 2015. The scarcity of studies on the antimicrobial susceptibility of this capsular type of P. multocida and growing concern about the general increase of antimicrobial resistance mean that studies related to the performance of type B P. multocida against antibiotics are necessary to establish accurate treatments and to monitor antimicrobial resistances.
RESULTS
Seventy-six isolates of P. multocida type B from pigs and cattle with acute septicemia were tested for susceptibility to 10 different antimicrobials. Bovine isolates were susceptible to all the antibiotics we tested except for lincomycin (94.4% of isolates were resistant). However, the antimicrobials we tested were less effective against swine isolates, of which none were susceptible to lincomycin. Furthermore, 29.3% swine isolates were resistant to tetracycline, 27.6% to penicillin, 20.7% to oxytetracycline, 17.3% to chloramphenicol, 15.5% to gentamicin, and 3.4% to enrofloxacin; no resistance to ceftiofur was detected. No multidrug resistant isolates were detected from cattle, while 25.86% of swine isolates were resistant to three or more antibiotic classes.
CONCLUSIONS
In this study, the lower resistance rates and multidrug resistant isolates reported for P. multocida type B derived from cattle compared to those isolated from pigs may be related to the increased use of antibiotics in the porcine industry in Spain. Lincomycin is not recommended for the treatment of acute septicemia in pigs or cattle, rather, the use of ceftiofur, enrofloxacin, or gentamicin is indicated as an emergency treatment in the early stages of disease; once the susceptibility results are known, the use of tetracyclines, penicillin, or chloramphenicol should be prioritized. The increase in multidrug resistant isolates and antimicrobial resistance rates indicates that more attention should be paid to prevention as well as the responsible use of antibiotics.
Topics: Animals; Anti-Bacterial Agents; Cattle; Cattle Diseases; Drug Resistance, Bacterial; Drug Resistance, Multiple; Microbial Sensitivity Tests; Pasteurella Infections; Pasteurella multocida; Sepsis; Spain; Swine; Swine Diseases
PubMed: 32605567
DOI: 10.1186/s12917-020-02442-z -
Future Microbiology Aug 2010Over the past few decades, our understanding of the bacterial protein toxins that modulate G proteins has advanced tremendously through extensive biochemical and... (Review)
Review
Over the past few decades, our understanding of the bacterial protein toxins that modulate G proteins has advanced tremendously through extensive biochemical and structural analyses. This article provides an updated survey of the various toxins that target G proteins, ending with a focus on recent mechanistic insights in our understanding of the deamidating toxin family. The dermonecrotic toxin from Pasteurella multocida (PMT) was recently added to the list of toxins that disrupt G-protein signal transduction through selective deamidation of their targets. The C3 deamidase domain of PMT has no sequence similarity to the deamidase domains of the dermonecrotic toxins from Escherichia coli (cytotoxic necrotizing factor [CNF]1-3), Yersinia (CNFY) and Bordetella (dermonecrotic toxin). The structure of PMT-C3 belongs to a family of transglutaminase-like proteins, with active site Cys-His-Asp catalytic triads distinct from E. coli CNF1.
Topics: Bacterial Toxins; Bordetella; Cytotoxins; Deamination; Escherichia coli; GTP-Binding Proteins; Models, Biological; Pasteurella multocida; Protein Structure, Tertiary; Sequence Homology, Amino Acid; Signal Transduction; Yersinia
PubMed: 20722598
DOI: 10.2217/fmb.10.91 -
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 -
Journal of Bacteriology Nov 2009Pasteurella multocida is classified into 16 serotypes according to the Heddleston typing scheme. As part of a comprehensive study to define the structural and genetic...
Pasteurella multocida is classified into 16 serotypes according to the Heddleston typing scheme. As part of a comprehensive study to define the structural and genetic basis of this scheme, we have determined the structure of the lipopolysaccharide (LPS) produced by P. multocida strains M1404 (B:2) and P1702 (E:5), the type strains for serotypes 2 and 5, respectively. The only difference between the LPS structures made by these two strains was the absence of a phosphoethanolamine (PEtn) moiety at the 3 position of the second heptose (Hep II) in M1404. Analysis of the lpt-3 gene, required for the addition of this PEtn residue, revealed that the gene was intact in P1702 but contained a nonsense mutation in M1404. Expression of an intact copy of lpt-3 in M1404 resulted in the attachment of a PEtn residue to the 3 position of the Hep II residue, generating an LPS structure identical to that produced by P1702. We identified and characterized each of the glycosyltransferase genes required for assembly of the serotype 2 and 5 LPS outer core. Monoclonal antibodies raised against serotype 2 LPS recognized the serotype 2/5-specific outer core LPS structure, but recognition of this structure was inhibited by the PEtn residue on Hep II. These data indicate that the serological classification of strains into Heddleston serotypes 2 and 5 is dependent on the presence or absence of PEtn on Hep II.
Topics: Bacterial Proteins; Electrophoresis, Capillary; Ethanolamines; Immunoblotting; Lipopolysaccharides; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Molecular Structure; Pasteurella multocida; Serotyping; Spectrometry, Mass, Electrospray Ionization
PubMed: 19767423
DOI: 10.1128/JB.00787-09 -
Journal of Global Antimicrobial... Mar 2020To date, very few hypervirulent and multiantibiotic-resistant bacterial strains have been reported. This study reports the first hypervirulent and...
OBJECTIVES
To date, very few hypervirulent and multiantibiotic-resistant bacterial strains have been reported. This study reports the first hypervirulent and multiantibiotic-resistant Pasteurella multocida sequence type 342 (ST342) strain (GH161213) isolated from a Pekin duck in China.
METHODS
Minimum inhibitory concentrations (MICs) were determined according to Clinical and Laboratory Standards Institute (CLSI) guidelines (VET01-A4, 2013). Determination of the P. multocida GH161213 median lethal dose (LD) was determined in a mouse model and in ducklings. Plasmid pRCAD0338PM-1 was transferred to Escherichia coli J53 by conjugation. The whole genome sequence of P. multocida GH161213 was obtained using an Illumina HiSeq 2500 system. Antimicrobial resistance genes were analysed using the Comprehensive Antibiotic Resistance Database (CARD).
RESULTS
Pasteurella multocida GH161213 is a hypervirulent strain with an LD of <10 CFU in a mouse model and in ducklings. It also has a high level of multidrug resistance. Strain GH161213 contains a small conjugative plasmid harbouring the floR florfenicol resistance gene. It also contains multiple other antimicrobial resistance mechanisms.
CONCLUSION
The genome sequence of P. multocida GH161213 reveals a multidrug-resistant genotype. This is the first reported hypervirulent and multiantibiotic-resistant P. multocida strain.
Topics: Animals; Anti-Bacterial Agents; Bacterial Proteins; China; Disease Models, Animal; Drug Resistance, Multiple, Bacterial; Ducks; High-Throughput Nucleotide Sequencing; Lethal Dose 50; Mice; Microbial Sensitivity Tests; Pasteurella Infections; Pasteurella multocida; Plasmids; Virulence; Whole Genome Sequencing
PubMed: 31557567
DOI: 10.1016/j.jgar.2019.09.012 -
Microbial Genomics Mar 2022Fowl cholera caused by has re-emerged in Australian poultry production since the increasing adoption of free-range production systems. Currently, autogenous killed...
Fowl cholera caused by has re-emerged in Australian poultry production since the increasing adoption of free-range production systems. Currently, autogenous killed whole-cell vaccines prepared from the isolates previously obtained from each farm are the main preventative measures used. In this study, we use whole-genome sequencing and phylogenomic analysis to investigate outbreak dynamics, as well as monitoring and comparing the variations in the lipopolysaccharide (LPS) outer core biosynthesis loci of the outbreak and vaccine strains. In total, 73 isolates from two different free-range layer farms were included. Our genomic analysis revealed that all investigated isolates within the two farms (layer A and layer B) carried LPS type L3, albeit with a high degree of genetic diversity between them. Additionally, the isolates belonged to five different sequence types (STs), with isolates belonging to ST9 and ST20 being the most prevalent. The isolates carried ST-specific mutations within their LPS type L3 outer core biosynthesis loci, including frameshift mutations in the outer core heptosyltransferase gene () (ST7 and ST274) or galactosyltransferase gene () (ST20). The ST9 isolates could be separated into three groups based on their LPS outer core biosynthesis loci sequences, with evidence for potential phase variation mechanisms identified. The potential phase variation mechanisms included a tandem repeat insertion in and a single base deletion in a homopolymer region of . Importantly, our results demonstrated that two of the three ST9 groups shared identical rep-PCR (repetitive extragenic palindromic PCR) patterns, while carrying differences in their LPS outer core biosynthesis loci region. In addition, we found that ST9 isolates either with or without the tandem repeat insertion were both associated with a single outbreak, which would indicate the importance of screening more than one isolate within an outbreak. Our results strongly suggest the need for a metagenomics culture-independent approach, as well as a genetic typing scheme for LPS, to ensure an appropriate vaccine strain with a matching predicted LPS structure is used.
Topics: Australia; Cholera; Disease Outbreaks; Farms; Glycosyltransferases; Humans; Lipopolysaccharides; Pasteurella Infections; Pasteurella multocida; Phase Variation
PubMed: 35266868
DOI: 10.1099/mgen.0.000772 -
Iranian Biomedical Journal Jan 2021Pasteurella multocida is a Gram-negative, non-motile, non-spore forming, and aerobic/anaerobic cocobacillus known as the causative agent of human and animal diseases....
BACKGROUND
Pasteurella multocida is a Gram-negative, non-motile, non-spore forming, and aerobic/anaerobic cocobacillus known as the causative agent of human and animal diseases. Humans can often be affected by cat scratch or bite, which may lead to soft tissue infections and in rare cases to bacteremia and septicemia. Commercial vaccines against this agent include inactivated, live attenuated, and non-pathogenic bacteria. Current vaccines have certain disadvantages such as reactogenicity or reversion to virulence. Therefore, the aim of this study was to reach a multi-epitope vaccine candidate that could be serotype independent and covers most incident serotypes of P. multocida.
METHODS
In this study, reverse vaccinology strategy was used to identify potentially immunogenic and protective epitopes. First, multiple alignments of different sequences of Pasteurella lipoprotein E (PlpE) from various serotypes of P. multocida were analyzed to identify the conserved regions. Bioinformatics tools were then applied to predict and select epitopes for further studies.
RESULTS
Three different conserved immunogenic regions were selected according to the selected criteria, and their various sequential orders were evaluated structurally by in silico tools to find the best order.
CONCLUSION
In searching the epitopes of PlpE to design a new vaccine candidate against pasteurellosis, we found the region 1 + region 2 + region 3 (without any linker between regions) of epitope, including the regions of PlpE protein of P. multocida, as the appropriate serotype independent vaccine candidate against pasteurellosis.
Topics: Bacterial Outer Membrane Proteins; Bacterial Vaccines; Computational Biology; Computer Simulation; Epitope Mapping; Epitopes; Hydrophobic and Hydrophilic Interactions; Immunogenicity, Vaccine; Lipoproteins; Molecular Structure; Pasteurella Infections; Pasteurella multocida; Serogroup; Vaccines, Subunit
PubMed: 33129238
DOI: 10.29252/ibj.25.1.41