scholarly journals Use of ELISA to Detect Toxigenic Pasteurella Multocida in Atrophic Rhinitis in Swine

1992 ◽  
Vol 4 (4) ◽  
pp. 419-422 ◽  
Author(s):  
Terry L. Bowersock ◽  
Tom Hooper ◽  
Ronald Pottenger
Keyword(s):  
Tissue Culture ◽  
Pasteurella Multocida ◽  
Atrophic Rhinitis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cytotoxicity Assays ◽  
Highly Sensitive ◽  
Nasal Secretions ◽  
Toxigenic Strains ◽  
Swine Herds ◽  
Toxigenic Pasteurella Multocida

The use of an enzyme-linked immunosorbent assay (ELISA) as a means of detecting dermone-crotoxin-producing strains of Pasteurella multocida was investigated. The assay was evaluated as a means to identify toxigenic P. multocida isolates recovered from nasal secretions of swine with atrophic rhinitis. The sensitivity and specificity of the ELISA for detecting dermonecrotoxin-producing P. multocida strains were compared to those of mouse-inoculation and cytotoxicity assays. The ELISA was highly sensitive and more specific than animal inoculation or tissue culture assay and is thus a more effective method for screening swine herds for the presence of toxigenic strains of P. multocida. The ELISA is a rapid, effective, economical way to identify toxigenic P. multocida isolates.

scholarly journals Detection of Toxigenic Pasteurella Multocida Infections in Swine Herds by Assaying Antibodies in Sow Colostrum

1996 ◽  
Vol 8 (4) ◽  
pp. 455-459 ◽  
Author(s):  
K. Levonen ◽  
P. L. Frandsen ◽  
J. Seppänen ◽  
P. Veijalainen
Keyword(s):  
Pasteurella Multocida ◽  
Screening Method ◽  
Clinical Signs ◽  
Atrophic Rhinitis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Causative Organism ◽  
Serologic Screening ◽  
Swine Herds ◽  
Toxigenic Pasteurella Multocida ◽  
Pig Health

Toxigenic Pasteurella multocida is the causative agent of progressive atrophic rhinitis (PAR), a serious respiratory infection of swine. Diagnosis of the disease has hitherto been based on clinical signs, pathologic findings, and subsequent isolation of the agent. The best Finnish pig breeding herds participating in the Finnish Pig Health Scheme have been surveyed for PAR since 1963, and the disease has been eradicated from these herds. In this study, a total of 5,650 colostrum samples from 188 Finnish Pig Health Scheme herds were analyzed with a new serologic screening method: an enzyme-linked immunosorbent assay (ELISA) able to detect antibodies to the toxin of P. multocida (PMT). Although the herds had been continuously controlled for PAR, 1 herd with PMT antibodies was found. The positive reactions in the ELISA were confirmed by isolating the causative organism. The origin of the infection also appeared to be obvious. The serologic ELISA is a suitable method for the detection and screening of toxigenic P. multocida-infected pig herds.

scholarly journals Molecular Fingerprinting of Pasteurella Multocida Associated with Progressive Atrophic Rhinitis in Swine Herds

1994 ◽  
Vol 6 (4) ◽  
pp. 442-447 ◽  
Author(s):  
Ian A. Gardner ◽  
Rick Kasten ◽  
Graeme J. Eamens ◽  
Kurt P. Snipes ◽  
Randall J. Anderson
Keyword(s):  
Pasteurella Multocida ◽  
Uv Light ◽  
Type A ◽  
Atrophic Rhinitis ◽  
Capsular Type ◽  
Molecular Fingerprinting ◽  
Whole Cell ◽  
Type D ◽  
Progressive Atrophic Rhinitis ◽  
Swine Herds

Ninety-six nasal isolates of Pasteurella multocida from swine herds with progressive atrophic rhinitis were characterized by restriction endonuclease analysis (REA) of whole-cell DNA, ribotyping, and plasmid analysis. For REA, bacterial DNA was digested with SmaI and electrophoresed in 0.7% agarose, and fragments were visualized with UV light. For ribotyping, EcoRI-digested and electrophoresed restriction fragments of whole-cell DNA were transferred to nitrocellulose membranes, hybridized with γ-32P-labeled Escherichia coli ribosomal RNA, and visualized by autoradiography. Phenotypes of isolates were toxigenic capsular type D ( n = 51), nontoxigenic type D ( n = 28), nontoxigenic type A ( n = 16), and toxigenic type A ( n = 1). Plasmids of various sizes were evident in 92.2% and 17.9% of toxigenic and nontoxigenic D strains, respectively, but were absent from all type A strains. Among the 4 phenotypes, there were 17 REA profiles and 6 ribotypes. For 3 of 17 REA patterns, multiple ribotypes were evident, and several REA types were evident in 5 of 6 ribotypes. Thirty-seven isolates of toxigenic capsular type D from Australian herds were either SmaI type B or C and ribotype 2, whereas 14 toxigenic D isolates from the USA and other countries were more heterogeneous (7 REA types and 6 ribotypes). The fingerprinting results provided evidence in support of the hypothesis of a single source infection in Australia associated with the introduction of breeding pigs from overseas.

scholarly journals Two-Color Hybridization Assay for Simultaneous Detection of Bordetella bronchiseptica and Toxigenic Pasteurella multocida from Swine

1998 ◽  
Vol 36 (11) ◽  
pp. 3342-3346 ◽  
Author(s):  
Karen B. Register ◽  
Ruby M. Lee ◽  
Cindy Thomson
Keyword(s):  
Pasteurella Multocida ◽  
Simultaneous Detection ◽  
Experimental Studies ◽  
Clinical Signs ◽  
Bordetella Bronchiseptica ◽  
Hybridization Assay ◽  
Atrophic Rhinitis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Primary Isolation ◽  
Comparison Of The Results

Bordetella bronchiseptica and toxigenicPasteurella multocida are the etiologic agents of swine atrophic rhinitis. Methods currently used for their identification are time-consuming and suffer from a lack of sensitivity. We describe a colony lift-hybridization assay for detection of B. bronchiseptica and toxigenic P. multocida that can be performed with a single colony lift derived from a primary isolation plate without the need for pure subcultures of suspect bacteria. Membranes are hybridized simultaneously to probes derived from the B. bronchiseptica alcA gene and the P. multocida toxA gene. A multicolor development procedure permits sequential detection of bound probes. The assay was tested with 84 primary isolation plates generated from nasal swabs from swine with clinical signs of atrophic rhinitis. Comparison of the results from the colony lift-hybridization assay with those from conventional testing, based on a combination of colony morphology, biochemical reactions, mouse lethality, and enzyme-linked immunosorbent assay, indicated that the colony lift assay has superior sensitivity and comparable specificity. This technique has wide application for diagnostic and experimental studies.

scholarly journals Evaluation of Techniques for the Detection of Toxigenic Pasteurella Multocida Strains from Pigs

1998 ◽  
Vol 10 (2) ◽  
pp. 169-173 ◽  
Author(s):  
Jose Antonio Amigot ◽  
Montserrat Torremorell ◽  
Carlos Pijoan
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Pasteurella Multocida ◽  
Complete Agreement ◽  
Atrophic Rhinitis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serotype A ◽  
Field Isolates ◽  
Dermonecrotic Toxin ◽  
Positive Results

Recently acquired field isolates and archived isolates from our collection of Pasteurella multocida were analyzed for production of dermonecrotic toxin. Detection of the toxin was carried out using a fetal lung feline (FLF) cell line and a commercial enzyme-linked immunosorbent assay (ELISA) kit. The dermonecrotic toxin gene ( ToxA) was also detected using a polymerase chain reaction (PCR) technique. Results from the 3 methods were compared. Field isolates (group 1) came from a commercial herd that had clinical signs of atrophic rhinitis. Fifty-six (17.9%) strains were isolated from 312 nasal swabs. Thirty-five of these strains belonged to serotype A and the rest (21/56), although probably serotype D, were not characterized further. All of these strains were toxin negative based on both the ELISA and FLF cell culture results. Five isolates gave faint bands in the PCR reaction, and the rest (51/56) were PCR negative. PCR and ELISA were also performed from the initial swab cultures (mixed cultures); 7 samples gave faint PCR bands, but ELISA results were all negative. Archived strains (group 2) had been isolated from clinical cases of atrophic rhinitis and from cases of pulmonary pasteurellosis. A total of 76 strains were analyzed; 46 were serotype A, and the rest (30) were serotype D. ELISA and FLF cell culture tests were negative for all serotype A strains; however, 3 strains showed faint bands in the PCR reaction. Fourteen serotype D strains showed positive results in both the ELISA and the FLF cell culture tests. PCR from these samples also gave positive results showing a strong band in the gel. However, 4 strains that were ELISA and FLF cell culture negative showed a faint band in the PCR reaction. The 3 methods gave similar results in the detection of the P. multocida dermonecrotic toxin. However, complete agreement among the tests was achieved only when strong PCR bands were considered positive. This is the first report that demostrates the use of FLF cell line for the detection of toxigenic P. multocida.

Identification of toxigenic Pasteurella multocida in atrophic rhinitis of pigs by in vitro characterisation

1988 ◽  
Vol 65 (4) ◽  
pp. 120-123 ◽  
Author(s):  
GJ EAMENS ◽  
PD KIRKLAND ◽  
MJ TURNER ◽  
JA GARDNER ◽  
MP WHITE ◽  
...  
Keyword(s):  
Pasteurella Multocida ◽  
Atrophic Rhinitis ◽  
Toxigenic Pasteurella Multocida

scholarly journals Molecular characterization of Pasteurella multocida isolates from swine lungs by Randomly Amplified Polymorphic DNA

2015 ◽  
Vol 46 (1) ◽  
pp. 119-125
Author(s):  
Cristiane Silva Chitarra ◽  
Mayara Inácio Vincenzi da Silva ◽  
Laila Natasha Santos Brandão ◽  
Francielle Cristina Kagueyama ◽  
Stefhano Luis Candido ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Pasteurella Multocida ◽  
Economic Losses ◽  
Clinical Samples ◽  
Atrophic Rhinitis ◽  
Randomly Amplified Polymorphic Dna ◽  
Swine Industry ◽  
Mato Grosso ◽  
Swine Herds

ABSTRACT: Swine respiratory diseases such as atrophic rhinitis and bronchopneumonia caused by Pasteurella (P.) multocida cause important economic losses to the modern swine industry. The purpose of this study was to characterize P. multocida strains isolated from swine lungs by RAPD (Randomly Amplified Polymorphic DNA) to demonstrate their genetic diversity. Ninety-four samples of fragments from lungs with pneumonia and sixty one samples without pneumonia were collected in slaughterhouses in Mato Grosso during the period from December 2009 to March 2010. Clinical cases in 2012 and 2013 were also included in this study. Among the lung fragments with macroscopic lesions, without macroscopic lesions and clinical samples, 40.42%, 4.49% and 100% were positive for P. multocida, respectively. Bacterial identification culturing was confirmed by PCR (polymerase chain reaction) by means of the amplification of the gene kmt1. RAPD technique was performed for 46 isolates, and in every isolate, a total of 7 to 11 amplification bands were detected, composed of 8 clusters based on genetic similarity. Thus, treatment, control and preventive measures should consider the genetic diversity of P. multocida populations in swine herds in order to improve the development of new protocols to produce antimicrobials and vaccines.

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