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Articles by Efren Diaz-Aparicio
Total Records ( 4 ) for Efren Diaz-Aparicio
  Efren Diaz-Aparicio , Beatriz Arellano-Reynoso , Enrique Herrera Lopez , Marisela Leal Hernandez and Francisco Suarez-Games
  A dairy herd with 17% Brucellosis prevalence was vaccinated or re-vaccinated with RB51. Five groups of ten cows each were classified according to their response to serologic tests. Additionally, one group with infected animals and one with animals originating from a Brucellosis free herd were used as positive and negative controls. A Radial Immunodiffusion test (RID) was used to confirm results. Indirect ELISA was carried out with LPS of B. abortus 2308 (LPS-S) or LPS of B. abortus RB51 (LPS-R) as primary antigens and anti-IgG1, IgG2, IgM and IgA as secondary antigens. When LPS-S was used as antigen, vaccinated and re-vaccinated cows that were positive to card and Rivanol tests, showed an IgG1-type response. Animals that were infected mainly showed an IgG2 type response. When LPS-R was used, none of the animals showed a significant response. Immunized cows that had contact with the wild-type strain showed humoral immune response that was detected as positive serology tests. Therefore, in endemic zones confirmation tests such as RID, Rivanol with titers above 1:100, or ELISA with LPS-S as antigen and IgG2, are essential to differentiate between animals vaccinated with RB51, which have a transitory immune response, from those that are infected.
  Laura Garcia Celis , Jehieli Girela alvarez , Beatriz Arellano-Reynoso , Victor Tenorio Gutierrez , Francisco Aguilar Romero , Pedro Mejia Sanchez , Francisco Suarez-Guemes and Efren Diaz-Aparicio
  There are no assays of experimental infection of sheep with Histophilus somni and their immune response. In order to know the humoral immune response of sheep experimentally infected by respiratory tract with H. somni, 19 male, 6 month-old sheep were immuno-suppressed with dexamethasone, 1 day before being inoculated with Parainfluenza 3 virus and during 5 days after viral inoculation. Seven days after viral inoculation, one animal was slaughtered in order to observe lesions caused by the virus. Another 12 animals were inoculated intratracheally with H. somni (1109 UFC mL 1), leaving 6 animals as controls that received same pathway sterile PSS. Heart and respiratory rates were taken, as well as rectal temperature, during 14 days after challenge. Serum and nasal exudate samples were collected to determine IgG, IgM and IgA levels by indirect ELISA test, using H. somni 40 kDa outer membrane protein as antigen. Two sheep from the inoculated group and one from the control group were humanely euthanized every week during 6 weeks. Samples were collected from lung, tonsils, retropharyngeal and mediastinal lymph nodes; bacteriology studies were carried out in duplicate and DNA was extracted to perform PCR with primers designed for 16S ribosomal region in H. somni. Infected animals had temperature and respiratory rate increase, cough and mucopurulent nasal exudate; areas in the apical lung lobule were consolidated and there were adherences as well as. H. somni was isolated from only one animal from retropharyngeal node. Nevertheless, from nasal exudate samples, 11 of 12 infected sheep came out positive by PCR while the 6 non-infected controls were negative. ELISA test results for IgG were significantly different between infected and control animals at days 4 and 7 after inoculation, while no differences were found in IgM and IgA. It was concluded that in sheep experimentally infected, H. somni caused an IgG isotype humoral immune response. The presence of the bacteria could be detected by PCR in 91.66% of the animals. Even though the strain used in this research was previously passed through sheep, it did not cause lesions or signs that could suggest infection and it could not be recovered by bacteriology in most of the infected sheep.
  Marisela Leal-Hernandez , Enrique Salas Tellez , Alma Nunez Del Arco , Rodolfo A. Ugalde , Victor R. Tenorio-Gutierrez , Rigoberto Hernandez-Castro , Francisco Suarez Guemes and Efren Diaz-Aparicio
  The aim of this research was to evaluate the humoral and cellular response, produced in mice inoculated with Brucella ovis and mutants in virB10 and virB11, measuring IgG1, IgG2a, IgG2b and IgM levels and cytokines production IL-2, IL-4 e IFN-γ, in lymphocyte culture. Mice of 6-weeks-old Balb/c were immunized intraperitoneally with each strain of B. ovis and the mutants with 3x1013 CFU mL-1 in PBS. For the cellular response, they were injected 2 times. In the humoral response the predominant antibodies were IgG2b and IgM in the different strains. The cytokine production with the wild type were indicative of Th2 and low response of IFN-γ and IL-2, that is indicative of an answer Th1 type characteristic of intracellular bacteria. The Reo 198 strain, showed Th1 type response. The mutant ΔvirB11 only presented response to IFN-γ 24 h that decreases after 48 h and with the mutant virB10:: Gm the shown levels of the different cytokine were low, this is indicative that the genes virB10 and virB11 of B. ovis need are complete to induce a suitable immune response. In the virulence in mice only recovered the Reo and wild type strains until 1 week postinoculation and the mutants can recover until 4 and 5 weeks postinoculation. One concludes the immune response by the different strains of B. ovis was moderate and can be attributed to that the murine model not was the more recommended to evaluate to cellular response would presumably have been specific to specie by the poorly or inability response showed when were mice inoculated with the different strains of B. ovis.
  Maria Dolores Fuentes Delgado , Irene V. Vitela Mendoza , Beatriz Arellano-Reynoso , Rigoberto Hernandez Castro , Jose Francisco Morales Alvarez , Carlos Cruz-Vazquez , Francisco Suarez-Guemes and Efren Diaz-Aparicio
  The present study was aimed to detect B. abortus DNA from aborted cows and differentiated its vaccinal and/or field origin. The PCR technique was used to identify and differentiate S19, RB51and field strains. The research was performed in a dairy farm located in the state of Aguascalientes, Mexico, with reproductive problems and abortions. Cows were vaccinated when calves with RB51, normal dose and revaccinated with RB51reduced dose 8 months before starting the study. The number of revaccinations of each cow, was not known since it is customary to revaccinate in one or more occasions. Samples of milk, blood serum and vaginal exudates were collected from 30 cows that had aborted between day 166 and 260 of gestation, all the abortions occurred 22-30 days before sampling; Card, rivanol and radial immunodiffusion tests were performed for Brucellosis diagnosis. For the bacteriological study samples of milk and vaginal swabs were used. The isolated strains were subjected to PCR using specific primers for RB51, S19 and field strains. From the 30 vaginal exudate samples bacteriological analyzed, we isolated smooth B. abortus biotype 1 was isolated from four cows. Thirteen out of the 30 cows vaginal exudates analyzed by PCR revealed the presence of DNA corresponding to RB51 vaccine, the 13 cows that were positive to RB51 through PCR had negative results in both the bacteriological and serological studies.
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