Pathological Analysis of a Rabbit that Died from Pasteurellosis and Genetic Analysis of <i>Pasteurella multocida</i> serotype -:1 Isolated from the Case

ABSTRACT Temperature-sensitive (TS) plasmids were generated through chemical mutagenesis of a derivative of the streptomycin resistance parent plasmid pD70, isolated from Mannheimia hemolytica serotype 1. Three TS plasmids which failed to replicate at or above 42°C in M. hemolytica but which were fully functional below 31°C were selected for further analysis. Two of the TS plasmids were shown by sequencing to possess unique single-base-pair mutations. The third TS plasmid contained a unique base pair substitution and a second mutation that had been previously identified. These mutations were clustered within a 200-bp region of the presumed plasmid origin of replication. Site-directed single-nucleotide substitutions were introduced into the wild-type pD70 origin of replication to confirm that mutations identified by sequencing had conferred thermoregulated replication. Deletion analysis on the wild-type pD70 plasmid replicon revealed that approximately 720 bp are necessary for plasmid maintenance. Replication of the TS plasmids was thermoregulated in Pasteurella multocida and Haemophilus somnus as well. To consistently transform H. somnus with TS plasmid, in vitro DNA methylation with commercially available HhaI methyltransferase was necessary to protect against the organism's restriction enzyme HsoI (recognition sequence 5′-GCGC-3′) characterized herein.

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Molecular cloning of haemoglobin-binding protein HgbA in the outer membrane of Actinobacillus pleuropneumoniae

Microbiology ◽

10.1099/mic.0.27046-0 ◽

2004 ◽

Vol 150 (6) ◽

pp. 1723-1734 ◽

Cited By ~ 17

Author(s):

Ramakrishnan Srikumar ◽

Leonie G. Mikael ◽

Peter D. Pawelek ◽

Ali Khamessan ◽

Bernard F. Gibbs ◽

...

Keyword(s):

Outer Membrane ◽

Pasteurella Multocida ◽

Affinity Purification ◽

Pcr Amplification ◽

Actinobacillus Pleuropneumoniae ◽

Sole Source ◽

Genomic Libraries ◽

Iron Deficient ◽

Serotype 1 ◽

Internal Peptide

From the porcine pathogen Actinobacillus pleuropneumoniae cultivated in iron-deficient or haem-deficient media, haemoglobin (Hb)-agarose affinity purification was exploited to isolate an outer-membrane protein of ∼105 kDa, designated HgbA. Internal peptide sequences of purified HgbA were used to design oligonucleotide primers for PCR amplification, yielding amplicons that showed partial sequences with homology to hgbA of Pasteurella multocida. Upon screening two genomic libraries of A. pleuropneumoniae serotype 1 strain 4074, positive clones were assembled into an ORF of 2838 bp. HgbA (946 aa) includes a signal peptide of 23 aa and the deduced HgbA sequence (104 890 Da) also demonstrated a possible Ton box. The promoter region of hgbA from A. pleuropneumoniae serotype 1 showed consensus for −35 and −10 sequences and a putative Fur-binding site. RT-PCR confirmed that hgbA of A. pleuropneumoniae is upregulated in response to diminished levels of iron in the culture medium. While an internally deleted hgbA mutant was unable to use pig Hb as sole source of iron for growth, flow cytometry confirmed its Hb binding; the internally deleted sequences may not be required for Hb binding, but appear necessary for the iron supply from Hb. HgbA is required for growth of A. pleuropneumoniae in the presence of Hb as sole iron source.

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Genetic Analysis of Antibody Responses of Turkeys to Newcastle Disease Virus and Pasteurella multocida Vaccines

Poultry Science ◽

10.3382/ps.0731169 ◽

1994 ◽

Vol 73 (8) ◽

pp. 1169-1174 ◽

Cited By ~ 9

Author(s):

R.E. SACCO ◽

K.E. NESTOR ◽

Y.M. SAIF ◽

H.J. TSAI ◽

N.B. ANTHONY ◽

...

Keyword(s):

Genetic Analysis ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Pasteurella Multocida ◽

Antibody Responses

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USING AMPLIFIED FRAGMENT LENGTH POLYMORPHISM ANALYSIS TO DIFFERENTIATE ISOLATES OF PASTEURELLA MULTOCIDA SEROTYPE 1

Journal of Wildlife Diseases ◽

10.7589/0090-3558-44.2.209 ◽

2008 ◽

Vol 44 (2) ◽

pp. 209-225 ◽

Cited By ~ 5

Author(s):

David S. Blehert ◽

Keynttisha L. Jefferson ◽

Dennis M. Heisey ◽

Michael D. Samuel ◽

Brenda M. Berlowski ◽

...

Keyword(s):

Length Polymorphism ◽

Fragment Length ◽

Pasteurella Multocida ◽

Fragment Length Polymorphism ◽

Amplified Fragment Length Polymorphism ◽

Polymorphism Analysis ◽

Serotype 1

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Streptococclcs Suis Infection in Swine: A Retrospective Study of 256 Cases. Part II. Clinical Signs, Gross and Microscopic Lesions, and Coexisting Microorganisms

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879400600308 ◽

1994 ◽

Vol 6 (3) ◽

pp. 326-334 ◽

Cited By ~ 42

Author(s):

Rachel Y. Reams ◽

Lawrence T. Glickman ◽

Daniel D. Harrington ◽

H. Leon Thacker ◽

Terry L. Bowersock

Keyword(s):

Escherichia Coli ◽

Retrospective Study ◽

Pasteurella Multocida ◽

Bacterial Culture ◽

Clinical Signs ◽

Animal Disease ◽

Diagnostic Laboratory ◽

Serotype 1 ◽

The Central Nervous System ◽

Gross Lesions

A retrospective study of 256 cases of naturally acquired Streptococcus suis infections in swine submitted to the Indiana Animal Disease Diagnostic Laboratory from 1985 to 1989 was undertaken to describe the clinical signs, lesions, and coexisting organisms associated with S. suis serotypes 1–8 and 1/2. Infected pigs generally had clinical signs and gross lesions referable to either the respiratory system or to the central nervous system (CNS), but not both. Neurologic signs were inversely related to gross lesions in the respiratory tract ( R2 = −0.19, P = 0.003), as were respiratory signs and gross lesions in the CNS ( R2 = −0.19, P = 0.003). Suppurative bronchopneumonia was the most common gross lesion observed (55.2%, overall). Fibrinous and/or suppurative pleuritis, epicarditis, pericarditis, arthritis, peritonitis, and polyserositis were also reported. In 68% of the pigs, other bacteria in addition to S. suis were isolated. Escherichia coli (35.0%) and Pasteurella multocida (30.0%) were the most commonly recovered bacterial agents. Mycoplasma and viral agents were identified less often, and their role in the development of streptococcosis was difficult to assess. In pigs infected with serotypes 2–5, 7, 8, and 1/2, suppurative meningitis with suppurative or nonsuppurative encephalitis, suppurative bronchopneumonia, fibrinopurulent epicarditis, multifocal myocarditis, and cardiac vasculitis were the most common microscopic lesions observed, whereas pigs infected with serotype 1 generally presented with suppurative meningitis and interstitial pneumonia. Microscopic lesions were morphologically similar among serotypes and were also similar to those reported with other pyogenic bacteria. The distribution of clinical signs and the gross and microscopic lesions in pigs infected with S. suis varied among serotypes. However, these differences were not statistically significant and could not be used to distinguish between the various serotypes. These findings suggest that in pigs infected with S. suis, suppurative or fibrinopurulent inflammation in brain, heart, lungs, and serosae predominates and that bacterial culture is needed to confirm a diagnosis of streptococcosis in swine and to differentiate this disease from those caused by other pyogenic bacteria.

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Pasteurella multocida Serotype 1 Isolated from a Lesser Snow Goose: Evidence of a Carrier State

Journal of Wildlife Diseases ◽

10.7589/0090-3558-33.2.332 ◽

1997 ◽

Vol 33 (2) ◽

pp. 332-335 ◽

Cited By ~ 10

Author(s):

M. D. Samuel ◽

D. R. Goldberg ◽

D. J. Shadduck ◽

J. I. Price ◽

E. G. Cooch

Keyword(s):

Pasteurella Multocida ◽

Carrier State ◽

Snow Goose ◽

Lesser Snow Goose ◽

Serotype 1

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Antigenic analysis of Pasteurella multocida (serotype 1) by crossed immunoelectrophoresis: characterization of cytoplasmic and cell envelope associated antigens

Canadian Journal of Microbiology ◽

10.1139/m80-118 ◽

1980 ◽

Vol 26 (6) ◽

pp. 676-689

Author(s):

J. L. Bhasin ◽

L. Lapointe-Shaw

Keyword(s):

Pasteurella Multocida ◽

Cell Envelope ◽

Rabbit Serum ◽

Antigenic Analysis ◽

Whole Cells ◽

Crossed Immunoelectrophoresis ◽

Serotype 1 ◽

Determining Factor ◽

Envelope Antigen

The application of crossed immunoelectrophoresis to the analysis of a reference cytoplasm and cell envelope preparation from Pasteurella multocida serotype 1 revealed antigenic complexity not previously found. At least 55 cytoplasmic and 19 cell envelope antigens were clearly distinguished. Variation of anticytoplasm immunoglobulin concentration was a major determining factor in resolving the maximum array of cytoplasmic antigens.The use of intermediate gel modification of crossed immunoelectrophoresis permitted the recognition of antibodies in the preimmune rabbit serum against a number of cytoplasmic antigens and a single cell envelope antigen. This technique also demonstrated that reference cytoplasm obtained by 105 000 × g centrifugation of sonically disrupted pasteurellae and repeatedly washed reference cell envelope preparation contained antigens of either origin in amounts sufficient to elicit antibody responses in the host. Antisera to whole cells in the intermediate gel indicated that formalin killed P. multocida were capable of eliciting immune responses to both reference systems.

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STANDARDIZATION OF EFFECTIVE DOSE OF FOWL CHOLERA VACCINE IN PIGEON IN BANGLADESH

Bangladesh Journal of Veterinary Medicine ◽

10.3329/bjvm.v15i2.35518 ◽

2018 ◽

Vol 15 (2) ◽

pp. 97-105

Author(s):

M. T. Islam ◽

M. H. Ali ◽

A. Chandra ◽

S. Saha ◽

M. A. Islam

Keyword(s):

Effective Dose ◽

Antibody Titer ◽

Pasteurella Multocida ◽

Passive Hemagglutination ◽

Fowl Cholera ◽

Serotype 1 ◽

Group A ◽

The Mean ◽

And Control ◽

Different Doses

An experiment was conducted to determine the effective dose of formalin killed (FK) fowl cholera (FC) vaccines prepared with virulent avian Pasteurella multocida (PM 38) serotype 1 (X-73) collected from the laboratory of the Department of Microbiology and Hygiene, BAU, Mymensingh. To determine the effective dose of vaccine, 7 weeks old 30 pigeons were immunized and each group consists of 5 birds. The groups are represented by A, B, C, D, E and F. The birds belonging to groups (A-E) were vaccinated with different doses of vaccine, after two weeks of first, second immunization and challenge experiment, blood was collected from all vaccinated birds, and serum was analyzed to determine antibody titer against P. multocida by passive hemagglutination test (PHA). The PHA titer after two weeks of first vaccination were 16±3.92, 17.6±3.92, 25.6±3.92, 32±8.76, 35.2±7.84 of group A,B,C,D and E, respectively at the dose of 0.2ml (0.26×108 CFU)/birds, 0.4ml (0.5×108 CFU)/birds, 0.8 ml (1.04×108 CFU)/birds, 1ml (1.3×108 CFU)/birds, respectively. The PHA titer of prevaccination and control birds was <4. The PHA titer after 2 weeks of second vaccination or boostering were 32±8.76, 35.2±7.84, 44.8±7.84, 57.6±18.66, 70.4±15.68, of group A,B,C,D and E, respectively. After 2 weeks of challenge infection, the mean PHA titer were 44.8±7.84, 51.2±7.84, 70.4±15.68, 102.4±15.68 and 140.8±31.34 of group A,B,C,D and E, respectively. In this experiment, the antibody titer of the vaccinated pigeons with 0.4, 0.6, 0.8 and 1ml per bird via intramuscular route were higher than that of the pigeons vaccinated with 0.4ml/bird, 0.6ml/bird, 0.8ml/bird and 1ml/bird were satisfactory in terms of protective potential against P. multocida. For prevention and control of avian pasteurellosis 0.4ml to o.6ml (0.52×108 CFU to 0.78×108 CFU)/birds of vaccine may be used instead of 1ml (1.3×108 CFU)/birds for better immunization of pigeon against fowl cholera infection.

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Antigenic analysis of Pasteurella multocida (serotype 1) by crossed immunoelectrophoresis: characterization of whole cell associated antigens

Canadian Journal of Microbiology ◽

10.1139/m80-232 ◽

1980 ◽

Vol 26 (12) ◽

pp. 1392-1402

Author(s):

J. L. Bhasin ◽

L. Lapointe-Shaw

Keyword(s):

Pasteurella Multocida ◽

Cell Envelope ◽

Potassium Thiocyanate ◽

Antigenic Analysis ◽

Whole Cell ◽

Heat Stable ◽

Crossed Immunoelectrophoresis ◽

Serotype 1 ◽

Reference Preparation ◽

Antigen Antibody

Crossed immunoelectrophoresis and other related quantitative immunoelectrophoretic techniques have been used to elucidate the antigenic complexity of a reference preparation of capsular extract, potassium thiocyanate extract, lipopolysaccharide, heat-stable antigens, and free endotoxin from Pasteurella multocida serotype 1. The reactions of these cellular fractions, in crossed immunoelectrophoresis, with reference anti-whole cell immunoglobulins disclosed five antigens in the capsular extract, seven in the potassium thiocyanate extract, one to three in the lipopolysaccharide, three in the heat-stable antigens, and five in the free endoxin. Comparison of these reference antigen–antibody systems, in crossed immunoelectrophoresis, with intermediate gel containing either a reference anti-cell envelope or anticytoplasmic immunoglobulins not only revealed the presence of additional antigens but also gave insight into the probable cellular origins (i.e., cell surface, cell envelope, or cytoplasm) of various antigens unveiled by reference anti-whole cell immunoglobulins. Using the principle of tandem crossed immunoelectrophoresis and crossed-line immunoelectrophoresis the immunochemical relationships between the antigenic components of these reference antigen–antibody systems were established.

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Immunogenicity of potassium thiocyanate extracted and electrofocused Pasteurella multocida X-73I antigens in chickens and mice

Canadian Journal of Microbiology ◽

10.1139/m82-182 ◽

1982 ◽

Vol 28 (11) ◽

pp. 1219-1225 ◽

Cited By ~ 4

Author(s):

Kevin L. McKinney ◽

Paul A. Rebers ◽

Richard B. Rimler

Keyword(s):

Antibody Response ◽

Pasteurella Multocida ◽

Potassium Thiocyanate ◽

Particulate Fraction ◽

Supernatant Fraction ◽

Chicken Embryos ◽

Isoelectric Points ◽

Serotype 1 ◽

Soluble Supernatant

The immunogenicity of antigenic fractions obtained by the extraction of Pasteurella multocida strain X-73 (serotype 1) with potassium thiocyanate (KSCN) was determined in chickens and mice. The initial KSCN extract was centrifuged at 105 000 × g, and the antigens were separated into a particulate fraction (40p) and a soluble supernatant fraction (40s). The ultracentrifuged fractions were further resolved by preparative electrofocusing. The 40p fraction was resolved into two subgroups having isoelectric points of 3.5–3.9 and 5.5–6.0; the 40s fraction was resolved into five subgroups ranging in isoelectric points from 4.4 to 9.0. The 40p fractions were antigenically similar and contained lipopolysaccharide (LPS) and protein. The 40s fractions were antigenically distinct from the 40p fractions and from each other; they contained proteins and polysaccharides but no LPS. The 40p antigens were strongly immunogenic in mice and chickens, whereas the 40s antigens were weakly immunogenic in chickens and not immunogenic in mice. The incorporation of Freund's complete adjuvant increased the immunogenicity of the 40s antigens in chickens. The 40p antigens induced greater frequencies of serological responses in chickens than the 40s antigens as detected by counterimmunoelectrophoresis and immunodiffusion. This suggested that the increased protection associated with the 40p antigens may have been the result of better antibody response. The toxicity of all the fractions was evaluated by determination of lethality for 10-day-old chicken embryos because of the sensitivity and reliability of the test. The 40p fraction had an LD50 = 0.38 μg, and the 40s fraction had an LD50 = 2.5 μg. Since the 40s fraction contained no detectable LPS, it is likely that two toxins are present, one which contains LPS and one which does not.

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