Caracterización de cepas de campo de <i>Herpesvirus suino 1</i> aisladas en Argentina

Se caracterizaron cepas argentinas de herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%.The genomic characterization of Suid herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Facultad de Ciencias Veterinaria

Caracterización de cepas de campo de <i>Herpesvirus suino 1</i> aisladas en Argentina

Se caracterizaron cepas argentinas de herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%.The genomic characterization of Suid herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Facultad de Ciencias Veterinaria

Extended phylogeny of the equine arteritis virus sequences including South American sequences

Objective: To perform genetic analysis of the ORF5 of equine arteritis virus (EAV) may provide new insights into the genetic evolution and origin of the Argentinean EAV sequences. Methods: A total of 76 sequences were analyzed by neighbor joining (NJ), maximum parsimony and maximum likelihood algorithms. The analysis of the selective pressures was performed using the Tajima's test. Results: The trees showed similar topologies. Two clades were identified: the first clade was formed by strains isolated mainly from a donkey, whereas the second clade presented four large groups from different geographic regions exclusively from Equus caballus. In this clade, we identified a group formed by South African and another one by South American and European sequences. In the latter, the monophyletic group was formed by seven Argentinean sequences. In the NJ tree, we identified a group formed by six Argentinean sequences. The Tajima's test showed a D value of 1.73663, indicating that the sequences analyzed follow a neutral evolution model. Conclusion: We concluded that the Argentinean sequences have a paraphyletic origin and that the fixation of point mutation might follow the neutral model evolution; however, we identified purifying pressures that may be involved in the differentiation of the EAV sequences.Facultad de Ciencias Veterinaria

Extended phylogeny of the equine arteritis virus sequences including South American sequences

Objective: To perform genetic analysis of the ORF5 of equine arteritis virus (EAV) may provide new insights into the genetic evolution and origin of the Argentinean EAV sequences. Methods: A total of 76 sequences were analyzed by neighbor joining (NJ), maximum parsimony and maximum likelihood algorithms. The analysis of the selective pressures was performed using the Tajima's test. Results: The trees showed similar topologies. Two clades were identified: the first clade was formed by strains isolated mainly from a donkey, whereas the second clade presented four large groups from different geographic regions exclusively from Equus caballus. In this clade, we identified a group formed by South African and another one by South American and European sequences. In the latter, the monophyletic group was formed by seven Argentinean sequences. In the NJ tree, we identified a group formed by six Argentinean sequences. The Tajima's test showed a D value of 1.73663, indicating that the sequences analyzed follow a neutral evolution model. Conclusion: We concluded that the Argentinean sequences have a paraphyletic origin and that the fixation of point mutation might follow the neutral model evolution; however, we identified purifying pressures that may be involved in the differentiation of the EAV sequences.Facultad de Ciencias Veterinaria

Caracterización de cepas de campo de <i>Herpesvirus suino 1</i> aisladas en Argentina

Se caracterizaron cepas argentinas de herpesvirus suino tipo 1 (HVS-1) mediante corte con las enzimas de restricción BamHI, BstEII y XhoI. La presencia del tipo genómico II había sido reportada en la Argentina una sola vez y, hasta el momento, no se han informado nuevos casos. Este estudio reveló una homogeneidad de los sitios BamHI, acorde con el número y el tamaño de los fragmentos. La presencia del fragmento BamHI #7 en los aislamientos argentinos analizados sugiere que éstos pertenecen al tipo salvaje (wild-type). Además, se caracterizó el principal dominio inmunogénico de la glicoproteína E de todas las cepas argentinas y se lo comparó con los de las cepas de referencia y con las secuencias disponibles en la base de datos GenBank. Los porcentajes de similitud obtenidos oscilaron entre 99 y 100%.The genomic characterization of Suid herpesvirus 1 (SHV-1) isolates from Argentina was accomplished by restriction pattern analysis using the BamHI, BstEII and XhoI enzymes. Type II genome has been described only once in Argentina. This study revealed considerable homogeneity of BamHI endonuclease sites in all the strains analyzed, according to the number and size of the fragments. No deletion of BamHI fragment #7 among the Argentinean isolates suggests that these strains are wild-type. In addition, the main antigenic domain of glycoprotein E of all the Argentinean strains, as well as the reference strains and sequences available in the GenBank, were characterized. The similarity percent oscillated between 99 and 100%.Facultad de Ciencias Veterinaria

Detection of equine herpesvirus 1 genome 1B in Argentina

To determine the genomic variation of equine herpesviruses (EHVs) isolated in Argentina between 1979 and the first half of 2004, DNA sequences from all 69 strains isolated were analysed. Sixty strains were recovered from aborted fetuses, one from leucocyte-rich plasma from a horse with respiratory signs and eight from cases of neonatal disease. The DNA was extracted from rabbit kidney epithelial (RK13) cells infected with each strain and digested with three restriction endonucleases (BamHI, Bg/II and KpnI). Two strains could be differentiated using BamHI restriction and were assigned to the EHV-1 1B prototype group. Only one of these two strains was typed EHV-1 1B with Bg/II. DNA digestion with KpnI was ineffective. The results obtained in this study demonstrate that the EHV-1 1B genome has been present in Argentina since at least 1996. The finding of two strains with this electropherotype suggests that there is genomic heterogeneity among Argentinian isolates.Facultad de Ciencias Veterinaria

Extended phylogeny of the equine arteritis virus sequences including South American sequences

Objective: To perform genetic analysis of the ORF5 of equine arteritis virus (EAV) may provide new insights into the genetic evolution and origin of the Argentinean EAV sequences. Methods: A total of 76 sequences were analyzed by neighbor joining (NJ), maximum parsimony and maximum likelihood algorithms. The analysis of the selective pressures was performed using the Tajima's test. Results: The trees showed similar topologies. Two clades were identified: the first clade was formed by strains isolated mainly from a donkey, whereas the second clade presented four large groups from different geographic regions exclusively from Equus caballus. In this clade, we identified a group formed by South African and another one by South American and European sequences. In the latter, the monophyletic group was formed by seven Argentinean sequences. In the NJ tree, we identified a group formed by six Argentinean sequences. The Tajima's test showed a D value of 1.73663, indicating that the sequences analyzed follow a neutral evolution model. Conclusion: We concluded that the Argentinean sequences have a paraphyletic origin and that the fixation of point mutation might follow the neutral model evolution; however, we identified purifying pressures that may be involved in the differentiation of the EAV sequences.Facultad de Ciencias Veterinaria

Equine arteritis virus: a new isolate from the presumable first carrier stallion in Argentina and its genetic relationships among the four reported unique Argentinean strains

Equine arteritis virus (EAV) was isolated from a testicle of the presumable first stallion infected with EAV in Argentina. This virus isolate (named LT-LP-ARG) was confirmed by GP5-specific PCR and indirect immunofluorescence assays. The PCR product was sequenced, and the phylogenetic analysis revealed that the LT-LP-ARG strain of EAV forms a monophyletic group, together with other strains previously isolated in our laboratory (LP02 group). However, all Argentinean EAV strains belong to a polyphyletic group. We believe that the virus isolate presented in this report could be the origin of EAV infection in our country.Facultad de Ciencias VeterinariasInstituto de Genética Veterinari

Detection of equine herpesvirus 1 genome 1B in Argentina

To determine the genomic variation of equine herpesviruses (EHVs) isolated in Argentina between 1979 and the first half of 2004, DNA sequences from all 69 strains isolated were analysed. Sixty strains were recovered from aborted fetuses, one from leucocyte-rich plasma from a horse with respiratory signs and eight from cases of neonatal disease. The DNA was extracted from rabbit kidney epithelial (RK13) cells infected with each strain and digested with three restriction endonucleases (BamHI, Bg/II and KpnI). Two strains could be differentiated using BamHI restriction and were assigned to the EHV-1 1B prototype group. Only one of these two strains was typed EHV-1 1B with Bg/II. DNA digestion with KpnI was ineffective. The results obtained in this study demonstrate that the EHV-1 1B genome has been present in Argentina since at least 1996. The finding of two strains with this electropherotype suggests that there is genomic heterogeneity among Argentinian isolates.Facultad de Ciencias Veterinaria

Modelos experimentales para el estudio de la patogenia de la muerte embrionaria en tricomonosis bovina y herpesvirosis equina

Desde hace mucho tiempo, por razones prácticas, científicas y éticas, se han desarrollado modelos experimentales con animales de laboratorio para su aplicación en investigaciones biomédicas. Los modelos animales se definen como “organismos vivientes con una inherente adquisición natural a procesos patológicos inducidos o espontáneos que, de una u otra manera, semejan el mismo fenómeno ocurrido en el hospedador natural” (Márquez, 1997).\nLos animales de laboratorio son modelos muy convenientes y herramientas útiles para utilizar en el estudio de muchas enfermedades infecciosas. En el caso de medicina veterinaria, en especial cuando se trata de enfermedades de grandes animales, el uso del hospedador natural para estudiar aspectos patogénicos e inmunológicos de una infección, muchas veces se torna dificultoso, tanto por las inconveniencias que genera el manejo de estos animales como por el costo que implica.\nDebido al interés y la complejidad en el estudio de las enfermedades infecciosas, hay una búsqueda en la profundización de los conocimientos del proceso intrínseco de la inmunopatogenia que requiere de la creación de modelos animales alternativos a los hospedadores naturales. El objetivo es generar diseños experimentales confiables y reproducibles, desarrollables en medios controlados en espacios reducidos y con menor costo. Entre las múltiples ventajas que derivan de la utilización de modelos experimentales en el estudio de diferentes enfermedades, se mencionan las siguientes como las más importantes:\n• Conocer la historia natural de la enfermedad, cuya etiología, patogenia, sintomatología y evolución pueden mantenerse en condiciones experimentales, sin la influencia de factores extraños que la modifiquen.\n• Reproducir la enfermedad en forma experimental, casi a voluntad, lo que permite disponer de la casuística necesaria.\n• Realizar estudios fisiopatológicos, desarrollando nuevas técnicas diagnósticas para tal enfermedad.\n• Estudiar las enfermedades en animales endocriados lo que permite un amplio campo de investigación en inmunología, patología y genética, entre otras áreas (Cuba Caparó, 1982).\nEn la selección de la especie utilizada como modelo animal es importante tener en cuenta algunas características generales: a) que permita la transferencia de la información, b) bajo costo y disponibilidad permanentes, c) generalización de los resultados, d) facilidad y adaptabilidad a la manipulación experimental, e) que se pueda contar con un número de animales necesarios para realizar el experimento, f) tiempo de vida, edad en que se alcanza la adultez y generación del número de progenies necesarias en poco tiempo, g) consecuencias ecológicas e implicancias éticas de su uso (Klein, 2000).Incluye las palabras de presentación del proyecto a cargo del Dr. Carlos O. Scoppa