Background and Aim: Canine degenerative myelopathy (DM) is an autosomal recessive inherited disea... more Background and Aim: Canine degenerative myelopathy (DM) is an autosomal recessive inherited disease that affects different dog breeds. It has an invariably fatal outcome once the clinical symptoms begin. This study aimed to investigate the population behavior of the mutation superoxide dismutase 1 (SOD1) c.118: G˃A responsible for the high risk of developing DM in two populations of German Shepherd dogs from Uruguay and Paraguay.
Materials and Methods: A total of 158 German Shepherd dogs from Uruguay (n = 114) and Paraguay (n = 44) were analyzed. Genomic DNA was extracted from peripheral whole blood. The SOD1 c.118: G˃A mutation was identified by polymerase chain reaction-restriction fragment length polymorphism and subsequently validated using sequencing. Allelic and genotypic frequencies and Hardy–Weinberg equilibrium were calculated for both populations. The rate of clinical progression was evaluated in animals homozygous for the mutation.
Results: The frequencies of allele A associated with a higher risk of DM, were 0.15 and 0.23 in Paraguay and Uruguay, respectively. Paraguay’s population was found to be in Hardy–Weinberg equilibrium (p = 1.00), whereas the population of dogs from Uruguay deviated from equilibrium (p = 0.008). When comparing the populations, no significant difference was observed in the distribution of genotypes (p = 0.26). When evaluating the clinical progression rate, all animals aged >10 years showed clinical symptoms compatible with DM.
Conclusion: This study demonstrated for the first time the presence of the SOD1:c118 G>A mutation in German Shepherd dogs from Uruguay and Paraguay. The frequency detected in Uruguay was significant. Although the frequency was lower in Paraguay, the allele was present. This demonstrates the need to implement genotyping tests as part of a possible DM control program in both countries studied.
espanolEl perro Cimarron es la unica raza canina autoctona en Uruguay. Dentro de un programa de c... more espanolEl perro Cimarron es la unica raza canina autoctona en Uruguay. Dentro de un programa de conservacion y con objeto de conocer la variabilidad genetica de esta raza es importante el analisis del ADN y dentro por consiguiente la utilizacion de marcadores moleculares de ADN entre los que se encuentran los denominados RAPD. El objetivo del presente trabajo es evaluar una serie de 10 marcadores RAPD en muestras de ADN de perros cimarrones mediante la metodologia de PCR. En el 70% de los marcadores se logro amplificacion de loci. Los RAPD 401 y 403 mostraron un mayor numero de loci polimorfico (58.33%), con un indice de similitud del patron de bandas de S=0.79 y 0.86 respectivamente. Los valores obtenidos nos estarian indicando una homogeneidad genomica en la muestra analizada. La utilizacion de esta serie de RAPD en una muestra mayor de perros cimarrones asi como en otras razas nos permitiran realizar estudios comparativos de variabilidad poblacional EnglishThe Cimarron dog is the only canine breed native in Uruguay. In a conservation program of this breed is important to use molecular DNA markers named RAPD to know the genetic variability. The objective of this work is to evaluate a series of 10 RAPD markers in DNA samples of Cimarron dogs with the PCR metodology. There were loci amplification in 70 % of the markers. RAPD 401 and 403 had a higer number of polimorphic loci (58,33 %), with a band sharing frequency of S=0.79 and 0.86 respectively. This numbers would indicate us a genomic homogenity in the analized sample. The use of this RAPD series in a higer cimarron dog sample like en other breeds will let us to do comparative studies of variability in populations.
Fragile sites (FS) seem to play a role in genome instability and may be involved in karyotype evo... more Fragile sites (FS) seem to play a role in genome instability and may be involved in karyotype evolution and chromosome aberrations. The majority of common fragile sites are induced by aphidicolin. Aphidicolin was used at two different concentrations (0.15 and 0.30 µM) to study the occurrence of FS in the cattle karyotype. In this paper, a map of aphidicolin induced break points and fragile sites in cattle chromosomes was constructed. The statistical analysis indicated that any band with three or more breaks was significantly damaged (P < 0.05). According to this result, 30 of the 72 different break points observed were scored as fragile sites. The Pearson correlation test showed a positive association between chromosome length and the number of fragile sites (r = 0.54). On the contrary, 21 FS were identified on negative R bands while 9 FS were located on positive R bands. cattle / chromosome / fragile sites / aphidicolin
Summary- Twenty Uruguayan Holstein-Friesian cows were cytogenetically examined, focusing on Xq 3.... more Summary- Twenty Uruguayan Holstein-Friesian cows were cytogenetically examined, focusing on Xq 3.1 fragility manifestation (Fra Xq). Comparative culture media studies were carried out by simultaneously culturing blood samples in TC 199 and RPMI 1640 media. Taking into account that X chromosome aberrations may be interfering with fertility, the cattle population were also evaluated for their reproductive performance. One thousand metaphase spreads were studied in each culture media and showed normal chromosome constitution in most cells. Frequency of Xq fragility detected in TC 199 was 0.9 % while in RPMI 1640 it was 1.8 % (0.05> P> 0.01). According to reproductive performance, repeat breeders manifested 92 % of all Fra Xq detected. The differences in expression in both tissue culture media and the frequencies of the Xq fragility observed in cattle with reproductive problems are discussed. fragile X / cattle / cytogenetics / fertility Résumé- Étude préliminaire sur l’incidence ...
The Uruguayan Cimarron dog is the only canine breed native from Uruguay; it was finally recognize... more The Uruguayan Cimarron dog is the only canine breed native from Uruguay; it was finally recognized as a breed in early 2011 by the "Federation Cynologique Internationale". This has meant an increased interest in the breed by breeders, exhibitors and pet owners in general. This interest has led to a significant increase in the number of animals registered in the genealogical records of the race, from nine hundred and thirty animals in October 1997, to more than 6,000 today. Moreover, these factors have led to different analysis are performed in order to continue with the knowledge of the genetics of this breed. Among the studies currently underway are those related to pharmacogenetics. CYP2D15 is one of the genes that are more involved in the metabolism of many widely used drugs in veterinary medicine, being described different polymorphisms for it. With primers: forward: TCTCTCCCTCTGCCCCAGAG; reverse: CCCAGGCTGTCCATCCTCTC, different polymorphisms were detected for exon 7 o...
La produccion lechera se encuentra asociada a una buena aptitud reproductiva del rodeo, en teoria... more La produccion lechera se encuentra asociada a una buena aptitud reproductiva del rodeo, en teoria se deberia lograr obtener un ternero/vaca por ano con un Intervalo parto concepcion (IPC) menor a los 90 dias. En el Uruguay el IPC en rodeos de produccion lechera es largo debido a diversos factores entre los que principalmente se destaca una pobre deteccion de los celos y una perdida importante por mortalidad embrionaria (Cavestany et al., 2007). En bovinos uno de los factores de origen genetico asociados a mortalidad embrionaria es la inestabilidad en el cromosoma sexual X (sitios fragiles Xq24, Xq31) (Danielak-Czech y Slota., 2004). En dicha region se encuentra identificado un gen (Gen TRO) que codifica para una proteina trofinina que en humanos interviene en la union del blastocisto al epitelio endometrial del utero durante la implantacion embrionaria. Este gen en bovinos ha sido mapeado en la region de inestabilidad cromosomica del X (BTAXq2533) (Asai., 2004). Dado el poco conocim...
Resumen en: Bovine herpesvirus type 1 (BoHV-1) is the etiological agent of Infectious Bovine Rhin... more Resumen en: Bovine herpesvirus type 1 (BoHV-1) is the etiological agent of Infectious Bovine Rhinotracheitis (IBR). Depending on the genotype, it produces a wide var...
La deficiencia en la adhesión leucocitaria bovina (BLAD) y en la enzima arginosuccinato sintetasa... more La deficiencia en la adhesión leucocitaria bovina (BLAD) y en la enzima arginosuccinato sintetasa (Citrulinemia) son enfermedades de herencia autosómica recesiva que han sido descritas a nivel mundial en la raza Holando. El objetivo de este estudio fue optimizar e implementar una metodología de genotipado para la identificación de animales portadores de los alelos causantes de estas enfermedades en una población cohorte de terneras de recría de la raza Holando de la cuenca lechera de Cerro Largo. Las muestras de ADN de 190 terneras Holando fueron extraídas a partir de sangre fresca, siguiendo normas y protocolos del Banco de ADN de la Unidad de Biotecnología INIA Las Brujas. El genotipado fue realizado mediante análisis PCR-RFLP con las enzimas de restricción TaqI para BLAD y Eco47I (AvaII) para Citrulinemia. La confirmación del genotipado fue evaluada mediante secuenciación de los productos amplificados de ambas enfermedades. Se detectó la presencia del alelo mutante para BLAD en u...
Azacytidine (5–azaC) is a substance that inhibits the metilation of DNA, being used in cloning ex... more Azacytidine (5–azaC) is a substance that inhibits the metilation of DNA, being used in cloning experiments, nuclear transference transgenesis and in chromosomal fragility studies. The manifestation of diverse pathologies has been observed in domestic animals born from cloning experiments. These would be associated to an incorrect nuclear reprogramation related to epigenetic metilation phenomena of the DNA. In the present work an induction curve with different final concentrations of 5–azaC (0 uM, 5 uM, 10 uM, 20 uM, 50 uM and 70 uM) were carried out, with the objective of studying the effects on chromosome structure in bovine lymphocyte cultures. The macroculture technique was used for chromosomic analysis, from a blood sample of a male bovine (Bos taurus, Holstein Friesian). A total of 50 metaphases were analyzed from each culture. Significative differences (p<0.01) were found regarding the break points of chromosome X in the control culture with respect to the culture with high...
Bovine presents a complex genome due to high number of repetitive sequences involved in macro and... more Bovine presents a complex genome due to high number of repetitive sequences involved in macro and micro-rearrangements that are essential in the bovids evolution. However, it has a morphologically simple karyotype, with acrocentric autosomes (2n=60,XX and 60,XY). The Robertsonian translocation rob(1;29), which produces embryo mortality, has been widely described in different breeds of Bos taurus but the complexity of its mechanism and its consequences in chromatin changes, are still not clear. Clastogenic agents are used as an approach to study this rearrangement in chromatin structure. Particularly, the aphidicolin (APC) inhibits the eukaryotic DNA polymerase α during replication. This fact permit to identify regions rich in dCTP due to a competition and spreading of the enzyme. Lymphocyte macro culture of 2 female Uruguayan Creole cattle carriers of rob(1;29) were APC-inducted (0,3uM) in one cell cycle. A new break point, rob(1;29)p13/21 (relative distance: p2c/p1=0.45), is locate...
The reproductive disorders in domestic animals often are associated with chromosomal abnormalitie... more The reproductive disorders in domestic animals often are associated with chromosomal abnormalities. In recent years, the study of chromosomal complement of i...
La presencia de fragilidad en el cromosoma X del bovino fue relacionada con problemas reproductiv... more La presencia de fragilidad en el cromosoma X del bovino fue relacionada con problemas reproductivos, sindrome de bakfy en temeros y diversas alteraciones fenotipicas. Recientemente, estudios previos realizados en Holando Uruguayo (Holstein-Friesian) mostraron la presencia de un sitio fragil en la region Xq 3.1 (banda G negativa). En este trabajo, se presenta la frecuencia de expresion espontanea del Fra Xq3.1 en diferentes muestras de bovinos. Las muestras sanguineas se procesaron por la tecnica estandar de cultivo linfocitario en medio completo RPM11640. La realizacion del bandeo de replicacion RBG mostro una banda particular R positiva. La muestra se subdividio en: (A) 46 hembras de diferentes edades con problemas reproductivos (repetidoras de servicio, freemartins, anestro, aborto); (B) 15 hembras de diferentes edades con reproduccion normal; y (C) 8 machos (7 co-mellizos de hembras, 1 con paladar hendido). Se examinaron 2758 placas metafasicas de las cuales 82 presentaron Fra Xg...
El alto grado de adaptabilidad fitness y la variacion encontrada en el color del pelaje de los bo... more El alto grado de adaptabilidad fitness y la variacion encontrada en el color del pelaje de los bovinos Criollos Americanas justifica realizar un analisis genetico a efectos de ser comparados con sus razas ancestrales Ibericas. Los estudios citogeneticos de la unica reserva de bovinos Criollos del Uruguay han comenzado con la busqueda de marcadores citogeneticos. La caracterizacion citogenetica se realizo en 99 animales fenotipicamente normales (73 hembras, 26 machos) de una poblacion de 600. Se cultivo sangre periferica en medio completo Rf MI 1640. Entre los cariotipos normales (2n _ 80,XX. XY1 se encontraron dos relaciones madres/crias con la formula cromosomica 2n = 59/rob (1;29), confirmandose por bandeo cromosomico RBG. El modelo de bandeo C mostro un bloque de heterocromatina (HC) en la region proximal del brazo q de la rob(1;29). La expresion del sitio fragil Xg3.1 se determino en un 6 p, cien de las metafases analizadas (N=710). Se observo un cromosoma Y metacentrico en todo...
Background and Aim: Canine degenerative myelopathy (DM) is an autosomal recessive inherited disea... more Background and Aim: Canine degenerative myelopathy (DM) is an autosomal recessive inherited disease that affects different dog breeds. It has an invariably fatal outcome once the clinical symptoms begin. This study aimed to investigate the population behavior of the mutation superoxide dismutase 1 (SOD1) c.118: G˃A responsible for the high risk of developing DM in two populations of German Shepherd dogs from Uruguay and Paraguay.
Materials and Methods: A total of 158 German Shepherd dogs from Uruguay (n = 114) and Paraguay (n = 44) were analyzed. Genomic DNA was extracted from peripheral whole blood. The SOD1 c.118: G˃A mutation was identified by polymerase chain reaction-restriction fragment length polymorphism and subsequently validated using sequencing. Allelic and genotypic frequencies and Hardy–Weinberg equilibrium were calculated for both populations. The rate of clinical progression was evaluated in animals homozygous for the mutation.
Results: The frequencies of allele A associated with a higher risk of DM, were 0.15 and 0.23 in Paraguay and Uruguay, respectively. Paraguay’s population was found to be in Hardy–Weinberg equilibrium (p = 1.00), whereas the population of dogs from Uruguay deviated from equilibrium (p = 0.008). When comparing the populations, no significant difference was observed in the distribution of genotypes (p = 0.26). When evaluating the clinical progression rate, all animals aged >10 years showed clinical symptoms compatible with DM.
Conclusion: This study demonstrated for the first time the presence of the SOD1:c118 G>A mutation in German Shepherd dogs from Uruguay and Paraguay. The frequency detected in Uruguay was significant. Although the frequency was lower in Paraguay, the allele was present. This demonstrates the need to implement genotyping tests as part of a possible DM control program in both countries studied.
espanolEl perro Cimarron es la unica raza canina autoctona en Uruguay. Dentro de un programa de c... more espanolEl perro Cimarron es la unica raza canina autoctona en Uruguay. Dentro de un programa de conservacion y con objeto de conocer la variabilidad genetica de esta raza es importante el analisis del ADN y dentro por consiguiente la utilizacion de marcadores moleculares de ADN entre los que se encuentran los denominados RAPD. El objetivo del presente trabajo es evaluar una serie de 10 marcadores RAPD en muestras de ADN de perros cimarrones mediante la metodologia de PCR. En el 70% de los marcadores se logro amplificacion de loci. Los RAPD 401 y 403 mostraron un mayor numero de loci polimorfico (58.33%), con un indice de similitud del patron de bandas de S=0.79 y 0.86 respectivamente. Los valores obtenidos nos estarian indicando una homogeneidad genomica en la muestra analizada. La utilizacion de esta serie de RAPD en una muestra mayor de perros cimarrones asi como en otras razas nos permitiran realizar estudios comparativos de variabilidad poblacional EnglishThe Cimarron dog is the only canine breed native in Uruguay. In a conservation program of this breed is important to use molecular DNA markers named RAPD to know the genetic variability. The objective of this work is to evaluate a series of 10 RAPD markers in DNA samples of Cimarron dogs with the PCR metodology. There were loci amplification in 70 % of the markers. RAPD 401 and 403 had a higer number of polimorphic loci (58,33 %), with a band sharing frequency of S=0.79 and 0.86 respectively. This numbers would indicate us a genomic homogenity in the analized sample. The use of this RAPD series in a higer cimarron dog sample like en other breeds will let us to do comparative studies of variability in populations.
Fragile sites (FS) seem to play a role in genome instability and may be involved in karyotype evo... more Fragile sites (FS) seem to play a role in genome instability and may be involved in karyotype evolution and chromosome aberrations. The majority of common fragile sites are induced by aphidicolin. Aphidicolin was used at two different concentrations (0.15 and 0.30 µM) to study the occurrence of FS in the cattle karyotype. In this paper, a map of aphidicolin induced break points and fragile sites in cattle chromosomes was constructed. The statistical analysis indicated that any band with three or more breaks was significantly damaged (P < 0.05). According to this result, 30 of the 72 different break points observed were scored as fragile sites. The Pearson correlation test showed a positive association between chromosome length and the number of fragile sites (r = 0.54). On the contrary, 21 FS were identified on negative R bands while 9 FS were located on positive R bands. cattle / chromosome / fragile sites / aphidicolin
Summary- Twenty Uruguayan Holstein-Friesian cows were cytogenetically examined, focusing on Xq 3.... more Summary- Twenty Uruguayan Holstein-Friesian cows were cytogenetically examined, focusing on Xq 3.1 fragility manifestation (Fra Xq). Comparative culture media studies were carried out by simultaneously culturing blood samples in TC 199 and RPMI 1640 media. Taking into account that X chromosome aberrations may be interfering with fertility, the cattle population were also evaluated for their reproductive performance. One thousand metaphase spreads were studied in each culture media and showed normal chromosome constitution in most cells. Frequency of Xq fragility detected in TC 199 was 0.9 % while in RPMI 1640 it was 1.8 % (0.05> P> 0.01). According to reproductive performance, repeat breeders manifested 92 % of all Fra Xq detected. The differences in expression in both tissue culture media and the frequencies of the Xq fragility observed in cattle with reproductive problems are discussed. fragile X / cattle / cytogenetics / fertility Résumé- Étude préliminaire sur l’incidence ...
The Uruguayan Cimarron dog is the only canine breed native from Uruguay; it was finally recognize... more The Uruguayan Cimarron dog is the only canine breed native from Uruguay; it was finally recognized as a breed in early 2011 by the "Federation Cynologique Internationale". This has meant an increased interest in the breed by breeders, exhibitors and pet owners in general. This interest has led to a significant increase in the number of animals registered in the genealogical records of the race, from nine hundred and thirty animals in October 1997, to more than 6,000 today. Moreover, these factors have led to different analysis are performed in order to continue with the knowledge of the genetics of this breed. Among the studies currently underway are those related to pharmacogenetics. CYP2D15 is one of the genes that are more involved in the metabolism of many widely used drugs in veterinary medicine, being described different polymorphisms for it. With primers: forward: TCTCTCCCTCTGCCCCAGAG; reverse: CCCAGGCTGTCCATCCTCTC, different polymorphisms were detected for exon 7 o...
La produccion lechera se encuentra asociada a una buena aptitud reproductiva del rodeo, en teoria... more La produccion lechera se encuentra asociada a una buena aptitud reproductiva del rodeo, en teoria se deberia lograr obtener un ternero/vaca por ano con un Intervalo parto concepcion (IPC) menor a los 90 dias. En el Uruguay el IPC en rodeos de produccion lechera es largo debido a diversos factores entre los que principalmente se destaca una pobre deteccion de los celos y una perdida importante por mortalidad embrionaria (Cavestany et al., 2007). En bovinos uno de los factores de origen genetico asociados a mortalidad embrionaria es la inestabilidad en el cromosoma sexual X (sitios fragiles Xq24, Xq31) (Danielak-Czech y Slota., 2004). En dicha region se encuentra identificado un gen (Gen TRO) que codifica para una proteina trofinina que en humanos interviene en la union del blastocisto al epitelio endometrial del utero durante la implantacion embrionaria. Este gen en bovinos ha sido mapeado en la region de inestabilidad cromosomica del X (BTAXq2533) (Asai., 2004). Dado el poco conocim...
Resumen en: Bovine herpesvirus type 1 (BoHV-1) is the etiological agent of Infectious Bovine Rhin... more Resumen en: Bovine herpesvirus type 1 (BoHV-1) is the etiological agent of Infectious Bovine Rhinotracheitis (IBR). Depending on the genotype, it produces a wide var...
La deficiencia en la adhesión leucocitaria bovina (BLAD) y en la enzima arginosuccinato sintetasa... more La deficiencia en la adhesión leucocitaria bovina (BLAD) y en la enzima arginosuccinato sintetasa (Citrulinemia) son enfermedades de herencia autosómica recesiva que han sido descritas a nivel mundial en la raza Holando. El objetivo de este estudio fue optimizar e implementar una metodología de genotipado para la identificación de animales portadores de los alelos causantes de estas enfermedades en una población cohorte de terneras de recría de la raza Holando de la cuenca lechera de Cerro Largo. Las muestras de ADN de 190 terneras Holando fueron extraídas a partir de sangre fresca, siguiendo normas y protocolos del Banco de ADN de la Unidad de Biotecnología INIA Las Brujas. El genotipado fue realizado mediante análisis PCR-RFLP con las enzimas de restricción TaqI para BLAD y Eco47I (AvaII) para Citrulinemia. La confirmación del genotipado fue evaluada mediante secuenciación de los productos amplificados de ambas enfermedades. Se detectó la presencia del alelo mutante para BLAD en u...
Azacytidine (5–azaC) is a substance that inhibits the metilation of DNA, being used in cloning ex... more Azacytidine (5–azaC) is a substance that inhibits the metilation of DNA, being used in cloning experiments, nuclear transference transgenesis and in chromosomal fragility studies. The manifestation of diverse pathologies has been observed in domestic animals born from cloning experiments. These would be associated to an incorrect nuclear reprogramation related to epigenetic metilation phenomena of the DNA. In the present work an induction curve with different final concentrations of 5–azaC (0 uM, 5 uM, 10 uM, 20 uM, 50 uM and 70 uM) were carried out, with the objective of studying the effects on chromosome structure in bovine lymphocyte cultures. The macroculture technique was used for chromosomic analysis, from a blood sample of a male bovine (Bos taurus, Holstein Friesian). A total of 50 metaphases were analyzed from each culture. Significative differences (p<0.01) were found regarding the break points of chromosome X in the control culture with respect to the culture with high...
Bovine presents a complex genome due to high number of repetitive sequences involved in macro and... more Bovine presents a complex genome due to high number of repetitive sequences involved in macro and micro-rearrangements that are essential in the bovids evolution. However, it has a morphologically simple karyotype, with acrocentric autosomes (2n=60,XX and 60,XY). The Robertsonian translocation rob(1;29), which produces embryo mortality, has been widely described in different breeds of Bos taurus but the complexity of its mechanism and its consequences in chromatin changes, are still not clear. Clastogenic agents are used as an approach to study this rearrangement in chromatin structure. Particularly, the aphidicolin (APC) inhibits the eukaryotic DNA polymerase α during replication. This fact permit to identify regions rich in dCTP due to a competition and spreading of the enzyme. Lymphocyte macro culture of 2 female Uruguayan Creole cattle carriers of rob(1;29) were APC-inducted (0,3uM) in one cell cycle. A new break point, rob(1;29)p13/21 (relative distance: p2c/p1=0.45), is locate...
The reproductive disorders in domestic animals often are associated with chromosomal abnormalitie... more The reproductive disorders in domestic animals often are associated with chromosomal abnormalities. In recent years, the study of chromosomal complement of i...
La presencia de fragilidad en el cromosoma X del bovino fue relacionada con problemas reproductiv... more La presencia de fragilidad en el cromosoma X del bovino fue relacionada con problemas reproductivos, sindrome de bakfy en temeros y diversas alteraciones fenotipicas. Recientemente, estudios previos realizados en Holando Uruguayo (Holstein-Friesian) mostraron la presencia de un sitio fragil en la region Xq 3.1 (banda G negativa). En este trabajo, se presenta la frecuencia de expresion espontanea del Fra Xq3.1 en diferentes muestras de bovinos. Las muestras sanguineas se procesaron por la tecnica estandar de cultivo linfocitario en medio completo RPM11640. La realizacion del bandeo de replicacion RBG mostro una banda particular R positiva. La muestra se subdividio en: (A) 46 hembras de diferentes edades con problemas reproductivos (repetidoras de servicio, freemartins, anestro, aborto); (B) 15 hembras de diferentes edades con reproduccion normal; y (C) 8 machos (7 co-mellizos de hembras, 1 con paladar hendido). Se examinaron 2758 placas metafasicas de las cuales 82 presentaron Fra Xg...
El alto grado de adaptabilidad fitness y la variacion encontrada en el color del pelaje de los bo... more El alto grado de adaptabilidad fitness y la variacion encontrada en el color del pelaje de los bovinos Criollos Americanas justifica realizar un analisis genetico a efectos de ser comparados con sus razas ancestrales Ibericas. Los estudios citogeneticos de la unica reserva de bovinos Criollos del Uruguay han comenzado con la busqueda de marcadores citogeneticos. La caracterizacion citogenetica se realizo en 99 animales fenotipicamente normales (73 hembras, 26 machos) de una poblacion de 600. Se cultivo sangre periferica en medio completo Rf MI 1640. Entre los cariotipos normales (2n _ 80,XX. XY1 se encontraron dos relaciones madres/crias con la formula cromosomica 2n = 59/rob (1;29), confirmandose por bandeo cromosomico RBG. El modelo de bandeo C mostro un bloque de heterocromatina (HC) en la region proximal del brazo q de la rob(1;29). La expresion del sitio fragil Xg3.1 se determino en un 6 p, cien de las metafases analizadas (N=710). Se observo un cromosoma Y metacentrico en todo...
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Papers by S. Llambí
Materials and Methods: A total of 158 German Shepherd dogs from Uruguay (n = 114) and Paraguay (n = 44) were analyzed. Genomic DNA was extracted from peripheral whole blood. The SOD1 c.118: G˃A mutation was identified by polymerase chain reaction-restriction fragment length polymorphism and subsequently validated using sequencing. Allelic and genotypic frequencies and Hardy–Weinberg equilibrium were calculated for both populations. The rate of clinical progression was evaluated in animals homozygous for the mutation.
Results: The frequencies of allele A associated with a higher risk of DM, were 0.15 and 0.23 in Paraguay and Uruguay, respectively. Paraguay’s population was found to be in Hardy–Weinberg equilibrium (p = 1.00), whereas the population of dogs from Uruguay deviated from equilibrium (p = 0.008). When comparing the populations, no significant difference was observed in the distribution of genotypes (p = 0.26). When evaluating the clinical progression rate, all animals aged >10 years showed clinical symptoms compatible with DM.
Conclusion: This study demonstrated for the first time the presence of the SOD1:c118 G>A mutation in German Shepherd dogs from Uruguay and Paraguay. The frequency detected in Uruguay was significant. Although the frequency was lower in Paraguay, the allele was present. This demonstrates the need to implement genotyping tests as part of a possible DM control program in both countries studied.
Keywords: degenerative myelopathy, genetic disease, German Shepherd dog, superoxide dismutase 1 gene.
Materials and Methods: A total of 158 German Shepherd dogs from Uruguay (n = 114) and Paraguay (n = 44) were analyzed. Genomic DNA was extracted from peripheral whole blood. The SOD1 c.118: G˃A mutation was identified by polymerase chain reaction-restriction fragment length polymorphism and subsequently validated using sequencing. Allelic and genotypic frequencies and Hardy–Weinberg equilibrium were calculated for both populations. The rate of clinical progression was evaluated in animals homozygous for the mutation.
Results: The frequencies of allele A associated with a higher risk of DM, were 0.15 and 0.23 in Paraguay and Uruguay, respectively. Paraguay’s population was found to be in Hardy–Weinberg equilibrium (p = 1.00), whereas the population of dogs from Uruguay deviated from equilibrium (p = 0.008). When comparing the populations, no significant difference was observed in the distribution of genotypes (p = 0.26). When evaluating the clinical progression rate, all animals aged >10 years showed clinical symptoms compatible with DM.
Conclusion: This study demonstrated for the first time the presence of the SOD1:c118 G>A mutation in German Shepherd dogs from Uruguay and Paraguay. The frequency detected in Uruguay was significant. Although the frequency was lower in Paraguay, the allele was present. This demonstrates the need to implement genotyping tests as part of a possible DM control program in both countries studied.
Keywords: degenerative myelopathy, genetic disease, German Shepherd dog, superoxide dismutase 1 gene.