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ECOIMMMUNOLOGY OF SWINE BRUCELLOSIS AND STRATEGIES FOR ERADICATION IN NATURALLY INFECTED FARM FROM CASEROS DEPARTAMENT. ARGENTINA.

2015; Frontiers Media; Volume: 6; Linguagem: Inglês

10.3389/conf.fimmu.2015.05.00230

1664-3224

Delgado Guillermo, De La Torre Flor, Correa David, Schaer Juan, Peralta Leticia, Thompson Carolina, Pereyra Norma, Besso Roberto, Calle Daniela, Gualtieri Catalina, Aguirre Nerina, Torioni De Echaide Susana, Arestegui Mirta,

Brucella: diagnosis, epidemiology, treatment

Event Abstract Back to Event ECOIMMMUNOLOGY OF SWINE BRUCELLOSIS AND STRATEGIES FOR ERADICATION IN NATURALLY INFECTED FARM FROM CASEROS DEPARTAMENT. ARGENTINA. Guillermo Delgado1, Flor De La Torre1, David Correa1, Juan M. Schaer1, Leticia Peralta1, Carolina Thompson2, Norma Pereyra1, Roberto Besso1, Daniela S. Calle1, Catalina Gualtieri1, Nerina Aguirre2, Susana M. Torioni De Echaide2 and Mirta B. Arestegui1* 1 Facultad de Ciencias Veterinarias. Universidad Nacional de Rosario., Argentina 2 Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Rafaela, Argentina Brucellosis continues to be a major public and animal health problem in many regions of the world. In Argentina B. suis bv 1 is the specie most frequently isolated in human brucellosis. Its eradication remains as an interdisciplinary challenge and needs the proper understanding of the ecoimmunology of Brucella suis infection in domestic pigs in order to design and implement valid strategies of eradication. The aim of the present work was to study the ecoimmunology of Brucella spp. infection in pigs naturally infected with B. suis from infected herd, through the evaluation of the IgG1 and IgG2 isotypes of immunoglobulin, molecular techniques and segregation of the positive reactors. The study was conducted in swine farm, surveyed as positive for brucellosis using Buffered Agglutination Test (BPAT), from Caseros department in Southern Santa Fe province, Argentina. The starting sampling (S1) included serum from 49 adult pigs that were analyzed to determine the initial intrapredial seroprevalence. Once detected, the serological positive reactors were segregated from herd as part of traditional control and eradication strategies of brucellosis. Between March and November 2014, the infected farm was analyzed using different serological and molecular tests. The second sampling (S2) including 19 adult pigs, was carried out 4 months later from S1. New cases of brucellosis detected by serology in S2 were also segregated from the herd. The third and fifth sampling (S3, S5) included 10 adults pigs each and fourth (S4) 7 respectively. Blood samples were obtained with and without anticoagulant (0.01M sodium citrate) by aseptic puncture of the cranial vena cava and dispensed in two vials. Sera was collected by centrifugation at 1000 xg for 20 min. Serum and blood whole samples were labeled and stored at -20 ºC until assayed. BPAT and RBT, CFT, c-ELISA, and FPA tests were performed according to OIE. IgG1 and IgG2 isotypes were determined using i-ELISA according to OIE (2012) with the following modifications. Two monoclonal antibodies against swine IgG1 and IgG2* (1/1000) and a conjugate anti-mouse IgG – Peroxidase (Jackson®) (1/6000) were used. The ratio of IgG1 and IgG2 isotypes was calculated by dividing OD values for IgG1 by OD values for IgG2. DNA was extracted from blood samples using a standard phenol/chloroform/isoamyl alcohol method (Leal Klevezas et al, 1995). The Brucella spp. gDNA was identified using Polymerase Chain Reaction (PCR) for amplifying a specific region of the eri gene (Leal Klevezas et al., 1995; Sangari and Agüero, 1994), and the species-specific stable IS711 element, localized in the Brucella chromosome (Bricker and Halling, 1994). The OD mean of IgG1/ IgG2 isotypes were compared between pigs of S2 and S3 using GraphPad Prism (version 5), p<0.05 was considered significant. The intrapredial prevalence at S1 was 22.45% (11/49). At S2, after segregation of pigs serologically positive from the farm, brucellosis was detected by different serological tests and by PCR. More than 80% of pigs from this new infected farm showed levels ≥ 1000UI-CFT and ≥70%I-cELISA. At S3 no seropositive pigs were found, while PCR was able to identify Brucella spp., new cases of brucellosis were still detected in pigs of this farm. The ratio of IgG1 / IgG2 was variable in infected pigs (PCR) according to presence of antibodies (CFT or c-ELISA), mean (±SD) of IgG1/IgG2 ratio, was significantly different (p< 0.05) in seronegative pigs between S2 (0.73 ± 0.29) and S3 (0.43 ±0.15). At S2 in seropositive pigs was (1.82 ± 1.5). These differences probably correlated with the stage of infection in individual animals. The kinetics of antibody production and the type of antibodies produced were key factors for analyzing serological results correctly and may help to predict abortion when pregnant animals are infected and thus allow their elimination before congeners can be contaminated. PCR and segregation of the positive reactors, as described in this work can also predict whether infection persists in a population. The combination of serological tests, molecular techniques and segregation of the positive reactors could be important tools to evaluate the ecoimmunology of swine brucellosis for effective eradication in farms from south of Santa Fe province, in order to complement the drawbacks of the conventional detection methods. * Kindly provided by Dr Javier Dominguez Juncal, INIA Spain. References Bricker, B.J. Halling S.M., 1995. Enhancement of the Brucella AMOS PCR assay for differentiation of B. abortus vaccine strains S19 and RB51. J. Clin. Microbiol.33, 1640-1642. Leal-Klevezas, D.S., Martinez-Vázquez, I.O., López-Merino, A., Pablo, J., Martinez, S.,1995. Single Step PCR for detection of Brucella spp from blood and milk of infected animals. J. Clin. Microbiol. 33, 3087-3090. OIE, World Organization for Animal Health, 2012. Chapter. 2.8.5. Porcine brucellosis. In: Terrestrial Manual, 7th ed. Paris, France, 2, 1106-1112. Sangari, F.J., Agüero, J., 1994. Identification of Brucella abortus B19 vaccine strain by the detection of DNA polymorphism at the ery locus. Vaccine 12, 435–438. Keywords: Swine brucellosis, Ecoimmunology, IgG1/IgG2 isotypes, Brucella suis, Brucellosis eradication Conference: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015. Presentation Type: Poster Presentation Topic: Veterinary and Comparative Immunology Citation: Delgado G, De La Torre F, Correa D, Schaer JM, Peralta L, Thompson C, Pereyra N, Besso R, Calle DS, Gualtieri C, Aguirre N, Torioni De Echaide SM and Arestegui MB (2015). ECOIMMMUNOLOGY OF SWINE BRUCELLOSIS AND STRATEGIES FOR ERADICATION IN NATURALLY INFECTED FARM FROM CASEROS DEPARTAMENT. ARGENTINA.. Front. Immunol. Conference Abstract: IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología. doi: 10.3389/conf.fimmu.2015.05.00230 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 15 Apr 2015; Published Online: 14 Sep 2015. * Correspondence: Dr. Mirta B Arestegui, Facultad de Ciencias Veterinarias. Universidad Nacional de Rosario., Casilda, Santa Fe, 2170, Argentina, maresteg@hotmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Guillermo Delgado Flor De La Torre David Correa Juan M Schaer Leticia Peralta Carolina Thompson Norma Pereyra Roberto Besso Daniela S Calle Catalina Gualtieri Nerina Aguirre Susana M Torioni De Echaide Mirta B Arestegui Google Guillermo Delgado Flor De La Torre David Correa Juan M Schaer Leticia Peralta Carolina Thompson Norma Pereyra Roberto Besso Daniela S Calle Catalina Gualtieri Nerina Aguirre Susana M Torioni De Echaide Mirta B Arestegui Google Scholar Guillermo Delgado Flor De La Torre David Correa Juan M Schaer Leticia Peralta Carolina Thompson Norma Pereyra Roberto Besso Daniela S Calle Catalina Gualtieri Nerina Aguirre Susana M Torioni De Echaide Mirta B Arestegui PubMed Guillermo Delgado Flor De La Torre David Correa Juan M Schaer Leticia Peralta Carolina Thompson Norma Pereyra Roberto Besso Daniela S Calle Catalina Gualtieri Nerina Aguirre Susana M Torioni De Echaide Mirta B Arestegui Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. 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