Genetic characterization of Argentine and Bolivian Creole cattle breeds assessed through microsatellites

In the present report, the polymorphisms from 9 microsatellites were used to assess genetic diversity and relationships in 4 Creole cattle breeds from Argentina and Bolivia, 4 European taurine breeds, and 2 American zebu populations. The Creole populations display a relatively high level of genetic variation as estimated by allelic diversity and heterozygosity, whereas the British breeds displayed reduced levels of genetic diversity. The analysis of molecular variance indicated that 7.8% of variance can be explained by differences among taurine and zebu breeds. Consistent with these results, the first principal component (PC), which comprised the 40% of the total variance, clearly distinguishes these 2 groups. In addition, all constructed phylogenetic trees cluster together Nelore and Brahman breeds with robust bootstrap values. Only 1% of variance was due to difference between American Creole and European taurine cattle. Although this secondary split was supported by the classical genetic distance and the second PC (15%), the topology of trees is not particularly robust. The presence of zebu-specific alleles in Creole cattle allowed estimating a moderate degree of zebu admixture. When these data were compared with mitochondrial and Y chromosomal studies, a clearl pattern of male-mediated introgression was revealed. The results presented here contribute to the understanding of origin and history of the American Creole cattle.Instituto de Genética Veterinari

Genetic characterization of Argentine and Bolivian Creole cattle breeds assessed through microsatellites

In the present report, the polymorphisms from 9 microsatellites were used to assess genetic diversity and relationships in 4 Creole cattle breeds from Argentina and Bolivia, 4 European taurine breeds, and 2 American zebu populations. The Creole populations display a relatively high level of genetic variation as estimated by allelic diversity and heterozygosity, whereas the British breeds displayed reduced levels of genetic diversity. The analysis of molecular variance indicated that 7.8% of variance can be explained by differences among taurine and zebu breeds. Consistent with these results, the first principal component (PC), which comprised the 40% of the total variance, clearly distinguishes these 2 groups. In addition, all constructed phylogenetic trees cluster together Nelore and Brahman breeds with robust bootstrap values. Only 1% of variance was due to difference between American Creole and European taurine cattle. Although this secondary split was supported by the classical genetic distance and the second PC (15%), the topology of trees is not particularly robust. The presence of zebu-specific alleles in Creole cattle allowed estimating a moderate degree of zebu admixture. When these data were compared with mitochondrial and Y chromosomal studies, a clearl pattern of male-mediated introgression was revealed. The results presented here contribute to the understanding of origin and history of the American Creole cattle.Instituto de Genética Veterinari

Brote de mastitis por <i>Prototheca</i> spp. y estudio ambiental del agente

Las algas del género Prototheca son las únicas plantas que pueden causar enfermedades a los humanos y otros mamíferos, principalmente vacas lecheras, a través de una invasión activa y diseminación en los tejidos del hospedador. En este trabajo se reporta un brote de mastitis provocado por Prototheca en un tambo de la Cuenca Mar y Sierras y el posterior estudio del agente en el ambiente. En dicho tambo se detectaba Prototheca con baja frecuencia de aislamiento en leche de tanque. Entre noviembre y diciembre del 2020 se diagnosticó Prototheca en 14 casos de mastitis clínica. A mediados de diciembre se realizó un muestreo del ambiente del tambo y de materia fecal de 23 vacas para detectar las potenciales fuentes de infección y/o reservorios de Prototheca spp. Las muestras se sembraron en medio selectivo para Prototheca (Prototheca isolation medium, PIM). Se realizó un pre-enriquecimiento en caldo PIM y luego repiques en placas de agar PIM. Se analizó la morfología de las colonias y del microorganismo mediante coloraciones con azul de metileno.Trabajo publicado en Cagliada, Maria del Pilar Lilia y Galosi, Cecilia Mónica (comps.). I Congreso de Microbiología Veterinaria. Libro de resúmenes. La Plata: Facultad de Ciencias Veterinarias, 2021.Facultad de Ciencias Veterinaria

Analysis of genetic diversity and population structure in Argentine and Bolivian Creole cattle using five loci related to milk production

Data from five protein-coding loci related to dairy production were used to study the genetic diversity and population structure of Argentine and Bolivian Creole cattle breeds. Genomic DNA was extracted from blood samples of six Creole cattle breeds: Argentine (n = 230), Patagonian (n = 25); "Saavedreño" (n = 140), "Chaqueño Boliviano" (n = 30), "Yacumeño" (n = 27), and "Chusco" (n = 11). κ-casein, β-lactoglobulin, growth hormone and prolactin were measured by PCR-RFLP, while αS1-casein was typed by PCR-ASO. The results are discussed, focusing on: historical origin, recent differentiation and selection events, Zebu gene introgression, and population structure. This work shows that: (i) For the studied genes, the observed gene frequency profiles of Argentine and Bolivian Creole cattle breeds were close to the data reported for Iberian breeds and for other South-American Creole cattle breeds which are historically related; (ii) although Zebu gene introgression has been reported at the studied loci, these breeds seem to be far from the Zebu gene frequency profiles; and (iii) the Argentine and Bolivian Creole cattle showed significant levels of subdivision, but each population has maintained its degree of genetic variability.Facultad de Ciencias VeterinariasInstituto de Genética Veterinari

African and European mitochondrial haplotypes in South American Creole cattle

South American Creole cattle are direct descendants of the animals brought to the New World by the Spanish and Portuguese during the 16th century. A portion of the mitochondrial D-loop was sequenced in 36 animals from five Creole cattle populations in Argentina and four in Bolivia. Individuals belonging to the potentially ancestral Spanish breed Retinta were also analysed. Sequence comparisons revealed three main groups: two with the characteristics of European breeds and a third showing the transitions representative of the African taurine breeds. The African sequences were found in two populations from Argentina and three populations from Bolivia, whose only connections go back to colonial times. The most probable explanation for the finding is that animals could have been moved from Africa to Spain during the long-lasting Arabian occupation that started in the seventh century, and from the Iberian Peninsula to America eight centuries later. However, since African haplotypes were not found in the Spanish sample, the possibility of cattle transported directly from Africa cannot be disregarded.Facultad de Ciencias VeterinariasInstituto de Genética Veterinari

Assessing the association of single nucleotide polymorphisms in thyroglobulin gene with age of puberty in bulls

Puberty is a stage of sexual development determined by the interaction of many loci and environmental factors. Identification of genes contributing to genetic variation in this character can assist with selection for early pubertal bulls, improving genetic progress in livestock breeding. Thyroid hormones play an important role in sexual development and spermatogenic function. The objective of this study was to evaluate the association between single nucleotide polymorphisms (SNPs) located in thyroglobulin(TG) gene with age of puberty in Angus bulls. Four SNPs were genotyped in 273 animals using SEQUENOM technology and the association between markers and puberty age was analyzed. Results showed a significant association (P < 0.05) between these markers and puberty age estimated at a sperm concentration of 50 million and a progressive motility of 10%. This is the first report of an association of TG polymorphisms with age of puberty in bulls, and results suggest the importance of thyroidal regulation in bovine sexual development and arrival to puberty.Instituto de Genética Veterinari

Inferencia del origen del bovino criollo cubano a través del análisis de patri- y matrilinajes

Cattle was absent from America before the discovery. Initially, bovine were brought to Greater Antilles (La Española, Puerto Rico, Jamaica and Cuba islands), and in the course of a few years, they were taken from Caribbean islands to the rest of Latin America. Nowadays, Cuban Creole cattle population is about 1300 heads, mainly located in the eastern region of the island. With the aim of analyzing the maternal origin of Cuban Creole cattle and detect possible contemporaneous, male mediated, gene flow, a 240 pb fragment of mitochondrial D-loop (mtDNA) and five microsatellites of Y chromosome (BTY) were studied in 36 dams and 21 sires, respectively. Genetic diversity was evaluated through number of haplotypes, mean number of pairwise differences and nucleotide diversity. The phylogenetic analysis was performed using a median joining. A total of 15 mtDNA haplotypes were detected in the studied population (10 from the European haplogroup T3, 3 from the African T1, 1 from the Nearern East T2, and 1 ambiguous T1-T3). The number of polymorphic sites, the mean nucleotide diversity, and the mean number of pairwise differences were 23, 0.014 and 3.36, respectively. Two patrilinages were detected, both belonging to the Y3 Zebu haplogroup. In conclusion, Cuban Creole cattle population had a mtDNA haplotypic composition similar to the observed in Creole and Mediterranean breeds, what is in concordance with its historical origin. Y chromosome analysis evidenced a male mediated process of zebu introgression.Antes de descubrimiento, no existían bovinos en América. Los primeros, fueron introducidos en la Antillas Mayores (La Española, Puerto Rico, Jamaica y Cuba), y desde allí trasladados al resto de Latinoamérica. Actualmente, existen en Cuba alrededor de 1300 bovinos Criollos, concentrados principalmente en la región oriental. Con el objetivo de analizar el origen materno de esta raza y detectar eventos contemporáneos de flujo gènico por vía paterna, se analizó un fragmento de 240 pb del D-loop mitocondrial (mtADN) y 5 microsatélites del cromosoma Y (BTY), en 36 hembras y 21 machos respectivamente. La diversidad genética se estimó mediante el número de haplotipos, el número de sitios polimórficos, el número de diferencias nucleotídicas entre pares de secuencias y el índice de diversidad nucleotídica, mientras que el análisis filogenètico se realizó utilizando el método de median joining network. Dicho análisis permitió detectar 15 haplotipos mitocondriales (10 del haplogrupo europeo T3,3 del africano T1,1 del cercano oriente T2y 1 ambiguo T1-T3) y 3 haplotipos en el BTY, ambos del haplogrupo cebuíno Y3. En el mtADN se detectaron 23 sitios polimórficos con una diversidad nucleotídica de 0,014 y 3,36 diferencias medias entre pares de secuencias. En conclusión, la población de bovinos Criollos Cubanos presentó una composición haplotípica mitocondrial comparable a la de otras razas criollasy mediterráneas, hecho que concuerda con su origen histórico. El BTY evidenció altos niveles de introgresion paterna de genes del zebú.Instituto de Genética Veterinari

Analysis of genetic diversity and population structure in Argentine and Bolivian Creole cattle using five loci related to milk production

Data from five protein-coding loci related to dairy production were used to study the genetic diversity and population structure of Argentine and Bolivian Creole cattle breeds. Genomic DNA was extracted from blood samples of six Creole cattle breeds: Argentine (n = 230), Patagonian (n = 25); "Saavedreño" (n = 140), "Chaqueño Boliviano" (n = 30), "Yacumeño" (n = 27), and "Chusco" (n = 11). κ-casein, β-lactoglobulin, growth hormone and prolactin were measured by PCR-RFLP, while αS1-casein was typed by PCR-ASO. The results are discussed, focusing on: historical origin, recent differentiation and selection events, Zebu gene introgression, and population structure. This work shows that: (i) For the studied genes, the observed gene frequency profiles of Argentine and Bolivian Creole cattle breeds were close to the data reported for Iberian breeds and for other South-American Creole cattle breeds which are historically related; (ii) although Zebu gene introgression has been reported at the studied loci, these breeds seem to be far from the Zebu gene frequency profiles; and (iii) the Argentine and Bolivian Creole cattle showed significant levels of subdivision, but each population has maintained its degree of genetic variability.Facultad de Ciencias VeterinariasInstituto de Genética Veterinari

Development and in-house validation of a real-time polymerase chain reaction for the detection of Listeria monocytogenes in meat

Listeriosis is a foodborne disease caused by Listeria monocytogenes. The aims of this work were to develop and validate an in-house real-time polymerase chain reaction (RT-PCR) for the detection of L. monocytogenes, and to determine its prevalence in raw ground beef samples from 53 butcheries that also sell ready-to-eat foods. One set of primers and one hydrolysis probe were designed for hly gene detection and then challenged with pure strains. The detection was successful for all L. monocytogenes strains analyzed and negative for all non-L. monocytogenes strains (detection limit, 10 colony forming unit [CFU]/mL). Inclusivity, exclusivity, and analytical accuracy were 100%. L. monocytogenes was detected in 41.5% of raw ground beef samples from the 53 butcheries analyzed. This RT-PCR may be a valuable method for rapid detection of L. monocytogenes in meat.Instituto de Genética VeterinariaFacultad de Ciencias Veterinaria

Development and validation of two SYBR green PCR assays and a multiplex real-time PCR for the detection of Shiga toxin-producing Escherichia coli in meat

Shiga toxin-producing Escherichia coli (STEC) are recognized as food-borne pathogens. We developed and validated two SYBR green PCR (SYBR-PCR) and a real-time multiplex PCR (RT-PCR) to detect stx1 and stx2 genes in meat samples, and compared these techniques in ground beef samples from retail stores. One set of primers and one hydrolysis probe were designed for each stx gene. For RT-PCR, an internal amplification control (IAC) was used. All PCR intra-laboratory validations were performed using pure strains and artificially contaminated ground beef samples. A total of 50 STEC and 30 non-STEC strains were used. Naturally contaminated ground beef samples (n = 103) were obtained from retail stores and screened with SYBR-PCR and RT-PCR, and stxpositive samples were processed for STEC isolation. In the intra-laboratory validation, each PCR obtained a 1 × 102 CFU mL−1 limit of detection and 100% inclusivity and exclusivity. The same results were obtained when different laboratory analysts in alternate days performed the assay. The level of agreement obtained with SYBR-PCR and RT-PCR was kappa=0.758 and 0.801 (P b 0.001) for stx1 and stx2 gene detection, respectively. Two PCR strategies were developed and validated, and excellent performance with artificially contaminated ground beef samples was obtained. However, the efforts made to isolate STEC from retail store samples were not enough. Only 11 STEC strains were isolated from 35 stx-positive ground beef samples identically detected by all PCRs. The combination of molecular approaches based on the identification of a virulence genotypic profile of STEC must be considered to improve isolation.Facultad de Ciencias VeterinariasInstituto de Genética Veterinari