Seropositivity for Bovine Viral Diarrhea and Enzootic Bovine Leukemia viruses in Blanco Orejinegro cattle in Colombia and infection associated-factors / Soropositividade para o vírus da diarréia viral bovina e leucemia bovina enzoótica em bovinos Blanco Orejinegro na Colômbia e fatores associados à infecção

ABSTRACT Serological controls for diseases of viral origin in animal production systems and the identification of factors associated with infections are decisive elements to establish prevention and control measures. The aim of this study was to establish the serological status for Bovine Viral Diarrhea (BVD) and Enzootic Bovine Leukemia (EBL) viruses in Blanco Orejinegro (BON) cattle from Colombia, and to identify the factors associated with seropositivity. A cross-sectional study was conducted with a total of 498 animals of all age groups and physiological states of the BON breed were selected, belonging to 14 herds located in 6 states of Colombia, in which a survey with 27 questions was conducted. By means of the chi-square test, possible factors associated with seropositivity against the 2 viruses were identified. A sample of 4 ml of blood was taken from each animal to extract plasma and make indirect Elisa tests to detect antibodies against both pathogens. General seropositivity of 27,1% was obtained for EBL, finding as factors associated with seropositivity the inadequate disposition of placental tissues after delivery of the cows and the non-performance of serological tests on new animals entering the herd. For BVD, seropositivity obtained was 50,6%, and the factors associated with seropositivity identified were having had a history of the disease in the herd, and using semen from bulls that are not known to be free for the infection. We suggest establishing control measures considering the factors associated with each viral infection to limit their expansion in the BON cattle production systems of Colombia.

RESUMO Os controles sorológicos para doenças de origem viral nos sistemas de produção animal e a identificação de fatores associados à infecção são elementos decisivos para estabelecer medidas de prevenção e controle. O objetivo deste estudo foi estabelecer o status sorológico dos vírus da Diarréia Viral Bovina (BVD) e da Leucemia Bovina Enzoótica (EBL) em bovinos Blanco Orejinegro (BON) da Colômbia e identificar os fatores associados à soropositividade. Foram selecionados 498 animais de todas as faixas etárias e estados fisiológicos da raça BON, de 14 rebanhos localizados em seis regiões da Colômbia, nos quais foi realizada uma pesquisa para identificar possíveis fatores associados à soropositividade contra os dois vírus. Foi retirada uma amostra de 4 ml de sangue de cada animal para extrair plasma e fazer testes ELISA indiretos para detectar anticorpos contra os dois patógenos. Obteve-se soropositividade geral de 27,1% para EBL, encontrando como fatores associados à soropositividade a disposição inadequada dos tecidos placentários após o parto das vacas e a não realização de testes sorológicos em novos animais que entraram no rebanho. Para a BVD, a soropositividade obtida foi de 50,6%, e os fatores associados à soropositividade identificados foram: histórico de doença no rebanho e uso de sêmen de touros que não são reconhecidos como livres da infecção. Sugerimos o estabelecimento de medidas de controle considerando os fatores associados a cada infecção viral para limitar sua expansão nos sistemas de produção de gado BON da Colômbia.

Diversidad haplotípica del exón xii del gen dopamina p-hidroxilasa en tres razas bovinas / Haplothic diversity of exon xii of the dopamine p-hydroxylase gene in three cattle breeds

Los polimorfismos en el gen dopamina p-hidroxilasa (DBH) se asocian con el temperamento en los bovinos. En 16 animales de la raza criolla colombiana Blanco Orejinegro (BON), 20 Brahman (BRA) y 16 de Ganado de lidia (GLI), se evaluaron los polimorfismos del exón xii del gen DBH mediante secuenciación. Se encontraron 13 haplotipos, 6 en BON, 9 en BRA y 2 en GLI. El haplotipo H2 fue el más frecuente (0,615) y se encontró en las tres razas, mientras los haplotipos H6 y H9 fueron compartidos entre BON-BRA y BRA-GLI, respectivamente. La mayor diversidad genética (DG: 0,915 ± 0,04) y nucleotídica (DN: 0,011 ± 0,006) se encontró en la raza BRA, seguida por BON (DG: 0,700 ± 0,12; DN: 0,008 ± 0,004) y GLI (DG: 0,350 ± 0,14; DN: 0,0011 ± 0,001). Si bien el test de D-Tajima fue mayor que cero, no fue significativo (P > 0,05) en BON y BRA, pero en la raza GLI presentó un valor de -1,92 (P < 0,05). El análisis de varianza molecular mostró una variación entre las razas de 23,9% y una estructura poblacional (F^) de 0,23 (P < 0,001). Los mayores valores del coeficiente de coancestría se presentaron entre GLI y BRA (0,52) y entre GLI y BON (0,22). Se concluye que la raza GLI tiene baja diversidad en el gen DBH en comparación con las razas BON y BRA, que hay efectos de la selección natural en BON y BRA mientras que en la raza GLI se evidencia un barrido selectivo reciente y no a favor del temperamento.

Polymorphisms in the dopamine p-hydroxylase gene (DBH) have been associated with temperament in cattle. In 16 animals of Colombian creole breed Blanco Orejinegro (BON), 20 Brahman (BRA) and 16 fighting bulls (GLI), polymorphisms of exon xii of the DBH gene were evaluated by sequencing. 13 haplotypes were found, 6 in BON, 9 and 2 in BRA and GLI respectively. Haplotype H2 was the most frequent in the whole population (0,615) and it was found in all breeds. Haplotypes H6 and H9 were shared between BON-BRA and BRA-GLI, respectively. The greatest value of genetic (DG: 0.915 ± 0.04) and nucleotide (DN: 0.011 ± 0.006) diversity, was found in BRA, followed by BON (DG: 0.700 ± 0.12; DN: 0.008 ± 0.004) and GLI (DG: 0.350 ± 0.14; DN: 0.0011 ± 0.001). The D-Tajima test was greater than zero, but not significant (P > 0.05) in BON and BRA, GLI presented a value of -1.92 (P < 0.05). The analysis of molecular variance showed a variation between the breeds of 23.9% and a poblacional structure (F$t) of 0.23 (P < 0.001). The highest values of the coancestry coefficient were presented between GLI and BRA (0.52) and between GLI and BON (0.22). It is concluded that the GLI race has low diversity in the DBH gene compared to BON and BRA, that there are effects of neutral selection in BON and BRA breeds, whereas, in the GLI and not in favor of temperament.

Estimation of genetic parameters for age at first calving and calving interval in Blanco Orejinegro (BON) breed cattle populations in Colombia / Estimación de parámetros genéticos para edad al primer parto e intervalo entre partos en poblaciones bovinas de la raza Blanco Orejinegro (BON) en Colombia / Estimação de parâmetros genéticos para idade ao primeiro parto e intervalo de partos em populações bovinas da raça Blanco Orejinegro (BON) na Colômbia

Objective: to estimate genetic parameters for age at first calving (AFC) and calving interval (CI) in 22 herds of Blanco Orejinegro cattle in Colombia. Methods: a total of 1,256 records for AFC and 3,803 for CI, obtained between years 1981 and 2010 were analyzed. The (Co) variances components were estimated by a derivative-free restricted maximum likelihood procedure in a bi-trait animal model. Results: average of AFC and CI were 1,104 ± 141 and 487 ± 147 days, respectively. Heritabilities were 0.15 and 0.13 for AFC and CI, respectively, with -0.43 genetic correlation. The herd and year of birth were included as fixed effects for the AFC, while parity number and the covariate age of dam at farrow were analyzed for CI. All the effects had a significant influence over the CI variance. Conclusions: the values obtained for these traits indicate that selection for calving interval and age at first calving may have a relatively low impact, due to the large environmental effect on the variation of both parameters in these breed populations.

Objetivo: estimar los parámetros genéticos de la edad al primer parto (EPP) y del intervalo entre partos (IEP) en 22 poblaciones bovinas de la raza criolla colombiana Blanco Orejinegro. Métodos: se utilizaron 1.256 registros para EPP y 3.803 registros de IEP, obtenidos entre los años 1981 y 2010. Los componentes de (Co) varianza fueron estimados por máxima verosimilitud restringida libre de derivadas con un modelo animal bicaracterístico. Resultados: los promedios de la EPP y del IEP fueron de 1.104 ± 141 y 487 ± 147 días, respectivamente. Las heredabilidades obtenidas en el análisis fueron de 0.15 y 0.13 para EPP e IEP, respectivamente, con una correlación genética de -0.43. Se evaluaron los efectos fijos de año de nacimiento y hato para la EPP, también fue incluido el orden de parto y la covariable edad de la vaca al parto en el análisis del IEP, los cuales todos tuvieron una influencia significativa sobre la variación de este parámetro. Conclusiones: los valores obtenidos para estas características reproductivas en el presente estudio, indican que la selección para intervalo entre parto y edad al primer parto puede tener un efecto relativamente bajo, debido al amplio efecto ambiental sobre la variación de estos dos parámetros en las poblaciones de esta raza.

Objetivo: estimar os parâmetros genéticos de idade ao primeiro parto (EPP) e do intervalo entre partos (IEP) em 22 populações bovinas da raça crioula colombiana Blanco Orejinegro. Métodos: foram utilizadas 1.256 e 3.803 dados para IPP e IEP respectivamente, obtidos entre os anos 1981 e 2010. Os componentes de (Co) variância foram estimados por máxima verossimilhança restrita livre de derivadas com um modelo animal bicaracterístico. Resultados: as médias da EPP e IEP foram 1.104 ± 141 días (36.8 ± 4.7 meses) e 487 ± 147 días (16.2 ± 4.9 meses), respectivamente. As herdabilidades obtidas nas análises foram de 0.15 e 0.13 para EPP e IEP respectivamente, com uma correlação genética de -0.43. Foram incluídos os efeitos fixos de ano de parto e rebanho para EPP, também foi analisado o efeito de ordem de parto e a covariável de idade da vaca ao parto para o IEP. Todos os efeitos incluídos na análise foram significativos sobre a variação do IEP. Conclusões: os valores obtidos no presente estúdio para estas características reprodutivas indicam que a selecção para intervalo entre parto e idade ao primeiro parto podem ter um efeito relativamente baixo, devido ao amplio efeito ambiental sobre a variação destes dois parâmetros nas populações desta raça.

Methylation-specific PCR analysis in Col8A1 promoter in Creole cattle carrier of rob(1; 29)

Robertsonian translocation (rob(1;29)) is the most frequent structural chromosomal abnormality in cattle. Heterozygous carriers have a normal phenotype but show a 3-5 percent decrease in fertility. Chromatin decondensation was evaluated similar to the inactive X chromosome when submitted to demethylating agent. Based on this result, and the concept that imprinted genes are essential in embryonic development, we decided to query genes located on BTA1 and BTA29 that could undergo genome imprinting. The collagen typeVIII- alpha 1 (Col8A1) acted on extracellular matrix structural proteins. DNA bisulfite conversion and sequentiation methods were used to compare its differential methylation patterns. It was performed on eight Creole cattle DNA blood samples from normal and rob(1;29) carriers. An in silico screening for CpG islands in its promoter uncovered a single region of 454 bp prone to methylation. BiQ-Analizer software was used to show the selective conversion of unmethylated cytosines to uracils obtaining the following results: unmethylated CpGs: 0.000 (0 cases), methylated CpGs: 0.802 (77 cases) and CpGs not present: 0.198 (19 cases). No differences between samples were observed in this highly methylated region. This technique was successfully applied so it is a straightforward methodology that can be utilized to evaluate different tissue associated to specific gene expression.

Frecuencias Alélicas de Beta-lactoglobulina en ganado criollo Limonero / Allelic Frequency of Beta-lactoglobulin in Limonero Creole Cattle

Con el objeto de caracterizar el gen de la beta-lactoglobulina (BLG) en la raza Criollo Limonero, se utilizó la técnica PCR-RFLP en 163 animales puros de la estación local Carrasquero (Carrasquero-estado Zulia), los genotipos fueron determinados a través de electroforesis en geles de agarosa. Las frecuencias obtenidas del locus de la BLG fueron A (0,22) y B (0,78) y las frecuencias genotípicas fueron AA (0,07 ); AB (0,29) y BB (0,64), la población estudiada se encuentra en equilibrio de Hardy-Weinberg (P<0,05), los resultados indican que la frecuencia alélica del alelo B fue más alta que la de A, siendo esto importante, ya que se han determinado los efectos de esta variante alélica de la BLG sobre la cantidad de grasa y proteínas en la leche, la selección a favor del alelo B en la población conllevará a una mejora en la calidad y rendimiento en la producción de queso, estos resultados representan un valioso aporte al conocimiento de esta raza y de su importancia, ya que, representa una alternativa para sistemas dirigidos a la producción de queso.

In order to characterize the beta-lactoglobulina gene (BLG) in the Limonero Creole cattle through PCR-RFLP technique, 163 purebreed animals were used from the Carrasquero local station (Carrasquero-Zulia State), genotypes were determined through gel electroforesis in agarosa. Gene and genotypic frequencies obtained were A (0.22) and B (0.78) and AA (0.07), AB (0.24) and BB (0.64) respectively, the population is in equilibrium of Hardy-Weinberg with (P<0.05), the results indicate that the alelic frequency of was more high B but that A, being this important one, since the effects of the variants of the BLG on the amount of fat and proteins in milk have been determined, the selection in favor of allele B in the population will entail to an improvement in the quality and yield in the cheese production, these results represent a valuable contribution the knowledge of this race and its importance, since, represents an alternative for systems directed to the cheese production.

Polymorphism of locus DRB3.2 in populations of Creole Cattle from Northern Mexico

The polymorphism of locus BoLA-DRB3.2 of the Major Histocompatibility Complex was evaluated in two northern Mexican Creole cattle populations, Chihuahua (n = 47) and Tamaulipas (n = 51). The BoLA-DRB3.2 locus was typed by amplification and digestion with restriction endonuclease enzymes (PCR-RFLP). Fifty-two alleles were detected (28 previously reported and 24 new ones). In the Chihuahua population, 18 alleles and 5.5 effective alleles were found, while in the Tamaulipas population there were 34 and 10.8, respectively. The allele frequencies ranged from 0.011 to 0.383 in Chihuahua and from 0.010 to 0.206 in Tamaulipas. The frequencies of the new alleles in both cattle populations were low (0.010 to 0.053). The expected heterozygosity was 0.827 and 0.916, respectively, for the Chihuahua and Tamaulipas populations. Both populations presented a heterozygote deficit: [Chihuahua FIS = 0.1 (p = 0.019) and Tamaulipas FIS = 0.317 (p < 0.001)]. In conclusion, this study showed that the Mexican Creole cattle have many low-frequency alleles, several of which are exclusive to these populations. Genetic distances obtained show that the Mexican Creole cattle population is composed of independent populations, far apart from other South American Creole populations.

INTROGRESIÓN GENÉTICA DE Bos indicus (BOVIDAE) EN BOVINOS CRIOLLOS COLOMBIANOS DE ORIGEN Bos taurus / Genetic Introgression of Bos indicus (Bovidae) in Colombian Creole Cattle Bos taurus

El presente trabajo evidencia desde el punto de vista citogenético la introgresión genética, de origen paterno, de Bos indicus en ganado criollo colombiano descendiente de Bos taurus. Para este estudio se realizó el análisis cariológico de la morfología del cromosoma Y a partir de muestras de sangre heparinizada de 67 bovinos machos pertenecientes a siete razas criollas colombianas. Se reporta la presencia de cuatro ejemplares pertenecientes a la raza Romosinuano (40%) y 10 toros de la raza Casanareña (100%) con cromosoma Y de tipo acrocéntrico característico de Bos indicus, lo cual estaría evidenciando un alto grado de introgresión genética, en estas dos razas, posiblemente originada por la intensiva introducción de sementales de la raza Cebú en la ganadería criolla colombiana. En las otras cinco razas (Blanco Orejinegro (BON), Chino santandereano, Costeño con cuernos, Hartón del valle y Sanmartinero), los toros presentaron el cromosoma Y submetacéntrico, característico de Bos taurus.

This work evidenced, using a cytogenetics approach, that Bos indicus exerted a genetic introgression of paternal origin on Creole Colombian cattle descendent from Bos taurus. Analysis of chromosome Y morphology was carried out in heparinized blood samples of 67 bulls belonging to seven Colombian breeds. We report 4 sires belonging to the Romosinuano breed (40%) and 10 bulls of the Casanareño breed (100%) with acrocentric Y chromosome which is characteristic of Bos taurus. This finding indicates a high degree of genetic introgression in these two breeds probably caused by the continuous input of zebu stallions in the Colombian Creole breeds. In other five Creole breeds (Blanco Orejinegro -BON-, Chino Santandereano, Costeño con Cuernos, Hartón del Valle and Sanmartinero), the bulls had a submetacentric Y chromosome characteristic of Bos taurus.