16 research outputs found
Evaluation and Comparison of the Ability of Online Available Prediction Programs to Predict True Linear B-cell Epitopes
No full textThis work deals with the use of predictors to identify useful B-cell linear epitopes to develop immunoassays. Experimental techniques to meet this goal are quite expensive and time consuming. Therefore, we tested 5 free, online prediction methods (AAPPred, ABCpred, BcePred, BepiPred and Antigenic) widely used for predicting linear epitopes, using the primary structure of the protein as the only input. We chose a set of 65 experimentally well documented epitopes obtained by the most reliable experimental techniques as our true positive set. To compare the quality of the predictor methods we used their positive predictive value (PPV), i.e. the proportion of the predicted epitopes that are true, experimentally confirmed epitopes, in relation to all the epitopes predicted. We conclude that AAPPred and ABCpred yield the best results, as compared with the other programs and with a random prediction procedure. Our results also indicate that considering the consensual epitopes predicted by several programs does not improve the PPV.Fil: Costa, Juan Gabriel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; ArgentinaFil: Faccendini, Pablo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; ArgentinaFil: Sferco, Silvano Juan. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Física; ArgentinaFil: Lagier, Claudia Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Química Rosario; ArgentinaFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; Argentin
Latex-protein complexes from an acute phase recombinant antigen of Toxoplasma gondii for the diagnosis of recently acquired toxoplasmosis
No full textThe synthesis and characterization of latex-protein complexes (LPC), from the acute phase recombinant antigen P35 (P35Ag) of Toxoplasma gondii and "core-shell" carboxylated or polystyrene (PS) latexes (of different sizes and charge densities) is considered, with the aim of producing immunoagglutination reagents able to detect recently acquired Toxoplasmosis. Physical adsorption (PA) and chemical coupling (CC) of P35Ag onto latex particles at different pH were investigated. Greater amounts of adsorbed protein were obtained on PS latexes than on carboxylated latexes, indicating that hydrophobic forces govern the interactions between the protein and the particle surface. In the CC experiments, the highest amount of bound protein was obtained at pH 6, near the isoelectric point of the protein (IP = 6.27). At this pH, it decreased both the repulsion between particle surface and protein, and the repulsion between neighboring molecules. The LPC were characterized and the antigenicity of the P35Ag protein coupled on the particles surface was evaluated by Enzyme-Linked ImmunoSorbent Assay (ELISA). Results from ELISA showed that the P35Ag coupled to the latex particles surface was not affected during the particles sensitization by PA and CC and the produced LPC were able to recognize specific anti-P35Ag antibodies present in the acute phase of the disease.Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); ArgentinaFil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); ArgentinaFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral; ArgentinaFil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentin
Synthesis and characterization of carboxyl and acetal latexes by emulsion polymerization. Application to the production of latex-protein complexes for detecting chagas disease
No full textMonodisperse polymer particles with carboxyl and acetal functionalities were synthesized through a two-step emulsion polymerization process. In the first step, latex particles were synthesized by batch emulsion polymerization of styrene (St); and in the second step, the functional monomers (methacrylic acid or acrolein diethyl acetal) were copolymerized with St onto the previously formed polystyrene particles. The synthesized “core-shell” latexes were used as support for their sensitization (by covalent coupling) with two antigenic recombinant proteins of Trypanosoma cruzi (RP1 and CP1). Polymer latexes and latex-protein complexes were characterized by measuring their colloidal stability, average particle size, shell thickness and protein thickness through conductimetric titration, dynamic light scattering, turbidimetry and scanning electron microscopy.Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico Para la Industria Química; ArgentinaFil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico Para la Industria Química; ArgentinaFil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico Para la Industria Química; ArgentinaFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biologica; ArgentinaFil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico Para la Industria Química; Argentin
Immunoagglutination test to diagnose Chagas disease: comparison of different latex-antigen complexes
No full textOBJETIVE To evaluate the diagnostic performance of novel latex–protein complexes obtained from different antigens of Trypanosoma cruzi through immunoagglutination test using a panel of T. cruzipositive sera, leishmaniasis-positive sera and negative sera for both parasites.
METHODS Complexes’ behaviour using total parasite homogenate (TPH), two simple recombinant proteins (RP1 and RP5) and two chimeric recombinant proteins (CP1 and CP2) was comparatively evaluated. The area under ROC curves was used as an index of accuracy. Sensitivity, specificity and discrimination efficiency were assessed.
RESULTS All recombinant antigens showed higher specificity than TPH. The lower specificity of TPH was mainly due to cross-reacting peptides between T. cruzi and Leishmania spp. In turn, all performance indicators were higher for CP1 and CP2 than for RP1 and RP5. The carboxylated latex– CP2 (C2–CP2) complex was able to detect antibodies against T. cruzi. The values of area under ROC curve (0.96), sensitivity (92.3%, 95% CI: 79.4–100.0%) and specificity (84.0%, 95% CI: 67.6– 100.0%) indicate that the assay could be used as a screening test. conclusion The C2–CP2 complex could be an important tool to carry out sero-epidemiological studies.Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); ArgentinaFil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); ArgentinaFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; ArgentinaFil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentin
Synthesis of latex-antigen complexes from single and multiepitope recombinant proteins. Application in immunoagglutination assays for the diagnosis of Trypanosoma cruzi infection
No full textThe physical adsorption and the chemical coupling of recombinant proteins of Trypanosoma cruzi onto polystyrene and core-shell carboxylated particles were respectively investigated with the ultimate aim of producing latex-protein complexes to be used in an immunoagglutination assay able to detect the Chagas disease. To this effect, two single proteins (RP1 and RP5) and a multiepitope protein derived from three antigenic peptides (CP2) were evaluated, and sensitizations were carried out at different pHs. The maximum physical adsorption was produced at pHs close to the protein isoelectric point (i.e., pH 6 for RP5 and pH 5 for RP1 and CP2). High fractions of antigens were chemically bound to the carboxyl groups, and the highest surface density of linked protein was also observed at pHs close to the protein isoelectric point. The three latex-protein complexes obtained by covalent coupling at such pHs were tested with sera from a panel of 16 infected and 16 non-infected patients. In the immunoagglutination assays, the latex-CP2 complex produced the best discrimination between positive and negative sera.Fil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); ArgentinaFil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); ArgentinaFil: Caudana, Pamela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); ArgentinaFil: Vega, Jorge Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); ArgentinaFil: Marcipar, Ivan Sergio. Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe; ArgentinaFil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico Para la Industria Química (i); Argentin
Synthesis and Characterization of Latex-protein Complexes from Different Antigens of Toxoplasma gondii for Immunoagglutination Assays
No full textThe acute phase recombinant protein of Toxoplasma gondii P22Ag was expressed and purified and thehomogenate of the parasite was obtained from an infected mouse. These antigens were used to producelatex-protein complexes (LPC) through physical adsorption and chemical coupling onto different latexes,with the aim of producing immunoagglutination (IA) reagents able to detect recently acquiredtoxoplasmosis. Polystyrene and ?core-shell? latexes where employed, exhibiting varied particle size,functionality (carboxyl or epoxy), and charge density. In sensitization experiments for producing LPC, therecombinant protein showed better coupling efficiency onto the particles surface than the homogenateand this could be explained by the complex mixture of the homogenate, which includes a large numberof proteins of different molecular mass, isoelectric points, and hydrophobicity. The synthesized LPC wereemployed in IA assays. To this effect, the agglutination reaction was followed by measuring the changesin the optical absorbance by turbidimetry. Experiments against control sera were performed to evaluatethe performance of various LPC and it was observed that the IA test based on P22Ag and thecarboxylated latex of 350 nm of particle diameter allowed a good discrimination between acute sera andchronic/negative ones. The proposed test is cheap, rapid, and easy to implement.Fil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Costa, Juan Gabriel. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentin
Assessment of Cross-reactive Host-pathogen Antibodies in Patients With Different Stages of Chronic Chagas Disease
No full textTrypanosoma cruzi infection has been shown to induce humoral autoimmune
responses against host antigens tissues. Particularly, antibodies cross-reacting with myocardial antigens may play a role in the development of the severe forms of chronic Chagas disease. The aim of this study was to determine the association between clinical stage of the disease and the presence of autoantibodies in patients with chronic Chagasic disease.La infección por Trypanosoma cruzi induce una respuesta autoinmunitaria humoral contra diferentes antígenos del huésped. En especial, los anticuerpos que presentan reactividad cruzada con antígenos del miocardio tienen un papel importante en el desarrollo de las formas graves de la cardiopatía chagásica crónica. En este trabajo se analiza la asociación del estadio clínico de la enfermedad con la presencia de autoanticuerpos en pacientes con cardiopatía chagásica crónica.Fil: Vicco, Miguel Hernán. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Ferini, Facundo. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas;Fil: Rodeles Antonelli, Luz María. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Cardona, Paula. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; ArgentinaFil: Bontempi, Iván. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; ArgentinaFil: Lioi, Susana. Universidad Nacional de Rosario; ArgentinaFil: Beloscar, Juan. Universidad Nacional de Rosario; ArgentinaFil: Nara, Takeshi. Universidad de Juntendo; JapónFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; ArgentinaFil: Bottasso, Oscar Adelmo. Universidad Nacional de Rosario. Facultad de Ciencias Médicas. Instituto de Inmunología; Argentin
Desarrollo de un inmunógeno prototípico para el control de mastitis bovinas causadas por Staphylococcus aureus
No full textTesis para la obtención del grado académico de Doctora en Ciencias Biológicas, de la Universidad Nacional del Litoral, en 2012Staphylococcus aureus es el patógeno más frecuentemente aislado de casos de mastitis bovina en Argentina y el mundo. Las particulares características de las infecciones intramamarias por S. aureus, han llevado a la búsqueda de alternativas para coadyuvar al control de esta enfermedad, siendo una de ellas la inmunización. El objetivo general de este trabajo fue desarrollar una vacuna prototípica contra mastitis bovinas causadas por S. aureus. Se compararon las respuestas inmunes generadas en bovinos por bacterinas de S. aureus productoras de polisacárido capsular 5 (CP5), formuladas con Al(OH3), o Iscom Matrix. La formulación con Iscom Matrix potenció la producción de anticuerpos específicos en sangre y leche, con elevada capacidad opsónica. Posteriormente, se compararon las respuestas inmunes generadas en bovinos por una bacterina y un lisado de S. aureus CP5 formulados con Iscom Matrix. La formulación con lisado bacteriano mostró un mejor desempeño en cuanto a la producción de anticuerpos específicos y funcionales, y la expresión de citoquinas con potencial importancia en el control de infecciones por S. aureus. Finalmente, se obtuvo un inmunógeno prototípico compuesto por lisado de S. aureus CP5, suplementado con los antígenos recombinantes proteína de unión a fibronectina A (FnBP-A), clumpling factor A (ClfA) y β-toxina, formulado con Iscom Matrix. La inmunización de vaquillonas con el inmunógeno multicomponente potenció la respuesta de anticuerpos en sangre y leche dirigidos hacia los antígenos recombinantes. Los mismos resultaron eficientes en la opsonización del microorganismo y en la neutralización de la actividad de la β-toxina nativa de S. aureus in vitro.Staphylococcus aureus is the most frequently isolated pathogen from bovine mastitis in Argentina and worldwide. The peculiar pathogenic characteristics of S. aureus lead to the development of alternatives measures, like immunization, to control this disease. The aim of this study was to develop a prototype vaccine against S. aureus bovine mastitis.
Immune responses induced in heifers by S. aureus bacterins from a capsular polisacharide 5 (CP5) producing strain, formulated either with Al(OH)3 or Iscom Matrix™, were compared. Formulation of experimental immunogen with Iscom MatrixTM strengthened humoral in blood and milk, with an augmented opsonic capacity. In addition, immune responses generated in heifers by either a S. aureus CP5 bacterin or lysate, formulated with Iscom MatrixTM were evaluated. The bacterial lysate formulation showed a better performance regarding production of specific and functional antibodies and induction of cytokines involved in early immune responses to the immunogen. Finally, a prototype immunogen composed by S. aureus CP5 lysate, supplemented with recombinant antigens fibronectin binding protein A (FnBP-A), clumping factor A (ClfA) and β-toxin, adjuvanted with Iscom MatrixTM, was obtained. Heifers’ immunization with the multicomponent immunogen strengthened antibody responses against recombinant antigens in blood and milk. These antibodies were efficient in opsonizing S. aureus and blocking native β-toxin hemolytic activity in vitro.EEA RafaelaFil: Camussone, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentin
P35 and P22 Toxoplasma gondii antigens abbreviate regions to diagnose acquired toxoplasmosis during pregnancy: toward single-sample assays
No full textBackground: P35 and P22 Toxoplasma gondii proteins are recognized by specific IgG at the early infection stage, making them ideal for acute toxoplasmosis pregnancy control. Both proteins have been studied to discriminate between acute and chronic toxoplasmosis. However, results were hardly comparable because different protein obtainment procedures led to different antigens, the referencepanels used were not optimally typified, and avidity tests were either not performed or narrowly examined. Methods: We bioinformatically predicted P35 andP22 regions with the highest density of epitopes, and expressed them in pET32/BL21DE3 alternative expression system, obtaining the soluble proteins rP35a and rP22a. We assessed their diagnostic performance using pregnant woman serum samples typified as: not infected, NI (IgG−, IgM−), typical-chronic, TC (IgM−, IgG+), presumably acute, A (IgG+, IgM+, low-avidity IgG), and recentlychronic, RC (IgG+, IgM+, high-avidity IgG).Fil: Costa, Juan Gabriel. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Peretti, Leandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Garcia, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Peverengo, Luz. Universidad Nacional del Litoral; ArgentinaFil: Gonzalez, Veronica Doris Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Gugliotta, Luis Marcelino. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Dalla Fontana, María. Laboratorio Central de la Provincia de Santa Fe; ArgentinaFil: Lagier, Claudia Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; ArgentinaFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin
Immune response and functional role of antibodies raised in heifers against a Staphylococcus aureus CP5 lysate and recombinant antigens vaccine formulated with Iscom Matrix adjuvant
No full textStaphylococcus aureus is the most frequently isolated pathogen from bovine intramammary infections worldwide. Commercially available vaccines for mastitis control are composed either of S. aureus lysates or inactivated whole-cells formulated with traditional adjuvants. We recently showed the ability of a S. aureus CP5 lysate vaccine adjuvanted with Iscom Matrix to generate a longer lasting specific antibody response in blood and milk, with improved opsonic capacity, compared with a S. aureus CP5 whole-cell formulation. The aim of the present study was to obtain an experimental immunogen composed of lysed cells of a CP5 S. aureus strain supplemented with recombinant clumping factor A, fibronectin binding protein A and -toxin formulated with Iscom Matrix, characterize the immune response generated when immunizing pregnant heifers and assess the functional role of antibodies raised against this immunogen in experimental models. Both a lysate vaccine and a lysate + recombinant antigens vaccine elicited antibodies that promoted neutrophil phagocytosis and inhibited internalization into mammary epithelial cells, in vitro. Incorporation of defined antigenic molecules to the lysate formulation elicited a strong specific humoral immune response against both lysate and recombinant antigens and was associated with higher expression of regulatory and pro-inflammatory cytokines. In addition, antibodies were efficient for blocking S. aureus binding to bovine fibrinogen and fibronectin, and neutralizing -toxin effect in vitro, placing these antigens as candidates to be included in a formulation directed to prevent staphylococcal bovine mastitis.Fil: Camussone, Cecilia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Pujato, Nazarena. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Renna, Maria Sol. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Cs.veterinarias. Instituto de Ciencias Veterinarias del Litoral; ArgentinaFil: Veaute, Carolina Melania Isabel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; ArgentinaFil: Morein, B.. Uppsala University. Department of Clinical Virology; SueciaFil: Marcipar, Ivan Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Tecnología Inmunológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Calvinho, Luis Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Cs.veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentin
