Dataset Information

A Serological Survey on Swine Brucellosis Using Standard Procedures, Dot Blot, and Western Blot in Finisher Pigs in Central-North Italy.

ABSTRACT: In recent years, Brucella suis has been sporadically reported in Italy in domestic and wild swine. Since standard serological tests can determine false positive results, the development of alternative tests with improved sensitivity and specificity is rather essential. We analyzed 1212 sera collected at slaughterhouse from healthy pigs belonging to 62 farms of North-Central Italy. Sera were tested by Rose Bengal Test, Complement Fixation Test, and subsequently by a Dot Blot (DB) and Western Blot assays (WB). Only one serum resulted positive to all tests, indicating that swine brucellosis has a very limited spread. DB and WB could represent a support to the available serological tests; however, further studies to validate these tests are needed. In the presence of reemerging diseases, a prompt and continuous monitoring design is necessary to acquire epidemiological information for the subsequent application of specific health emergency plans.

SUBMITTER: Bertelloni F

PROVIDER: S-EPMC6313843 | biostudies-other | 2018 Oct

REPOSITORIES: biostudies-other

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A Serological Survey on Swine Brucellosis Using Standard Procedures, Dot Blot, and Western Blot in Finisher Pigs in Central-North Italy.

Bertelloni Fabrizio F Forzan Mario M Turchi Barbara B Sagona Simona S Mazzei Maurizio M Felicioli Antonio A Fratini Filippo F Cerri Domenico D

Veterinary sciences 20181003 4

In recent years, <i>Brucella suis</i> has been sporadically reported in Italy in domestic and wild swine. Since standard serological tests can determine false positive results, the development of alternative tests with improved sensitivity and specificity is rather essential. We analyzed 1212 sera collected at slaughterhouse from healthy pigs belonging to 62 farms of North-Central Italy. Sera were tested by Rose Bengal Test, Complement Fixation Test, and subsequently by a Dot Blot (DB) and Weste ...[more]

PMID: 30282958

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Project description:Western blotting has proven to be an important technique in analysis of receptor-ligand interactions (i.e., by ligand blotting) and for identifying molecules mediating cell attachment (i.e., by cell blotting). Conventional ligand blotting and cell blotting methods employ nondynamic (static) incubation conditions, whereby molecules or cells of interest are placed in suspension and overlaid on membranes. However, many cell-cell and cell-matrix adhesive interactions occur under fluid shear conditions, and shear stress itself mediates and/or facilitates the engagement of these physiologically appropriate receptors and ligands. Notably, shear forces critically influence the adhesion of circulating cells and platelets to vessel walls in physiologic cell migration and hemostasis, as well as in inflammatory and thrombotic disorders, cancer metastasis, and atherosclerosis. Use of nondynamic blotting conditions to analyze such interactions can introduce bias, overtly missing relevant effectors and/or exaggerating the relative role(s) of nonphysiologic adhesion molecules. To address this shortfall, we have developed a new technique for identifying binding interactions under fluid shear conditions, the "blot rolling assay." Using this method, molecules in a complex mixture are resolved by gel electrophoresis, transferred to a membrane that is rendered semi-transparent, and the membrane is then incorporated into a parallel-plate flow chamber apparatus. Under controlled flow conditions, cells or particles bearing adhesion proteins of interest are then introduced into the chamber and interactions with individual immobilized molecules (bands) can be visualized in real-time. The substrate molecule(s) supporting adhesion under fluid shear can then be identified by staining with specific antibodies or by excising the relevant band(s) and performing mass spectrometry or microsequencing of the isolated material. This method thus allows for the identification, within a complex mixture and without prior isolation or purification, of both known and previously uncharacterized adhesion molecules operational under dynamic conditions.

Dataset Information

A Serological Survey on Swine Brucellosis Using Standard Procedures, Dot Blot, and Western Blot in Finisher Pigs in Central-North Italy.

Publications

A Serological Survey on Swine Brucellosis Using Standard Procedures, Dot Blot, and Western Blot in Finisher Pigs in Central-North Italy.

Similar Datasets

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