Search

Brangus is a composite cattle breed developed with the objective of combining the advantages of Angus and Zebuine breeds (Brahman, mainly) in tropical climates. The aim of this work was to estimate breed composition both genome-wide and locally, at the chromosome level, and to uncover genomic regions evidencing positive selection in the Argentinean Brangus population/nucleus. To do so, we analysed marker data from 478 animals, including Brangus, Angus and Brahman. Average breed composition was 35.0% ± 9.6% of Brahman, lower than expected according to the theoretical fractions deduced by the usual cross-breeding practice in this breed. Local ancestry analysis evidenced that breed composition varies between chromosomes, ranging from 19.6% for BTA26 to 56.1% for BTA5. Using approaches based on allelic frequencies and linkage disequilibrium, genomic regions with putative selection signatures were identified in several chromosomes (BTA1, BTA5, BTA6 and BTA14). These regions harbour genes involved in horn development, growth, lipid metabolism, reproduction and immune response. We argue that the overlapping of a chromosome segment originated in one of the parental breeds and over-represented in the sample with the location of a signature of selection constitutes evidence of a selection process that has occurred in the breed since its take off in the 1950s. In this regard, our results could contribute to the understanding of the genetic mechanisms involved in cross-bred cattle adaptation and productivity in tropical environments.

Canine forms of osteoarthritis (OA) are very similar to those in humans and represent a welfare problem in the dog world population. In this study, we investigated the transcriptomic profile of peripheral blood in German Shepherd dogs with OA in order to identify putative diagnosis biomarkers. The bulk RNA-seq experiment was performed in a cohort of 12 adult dogs, 5 OA-affected and 7 unaffected. Radiographs of the affected dogs revealed signs of progressive OA in hip, elbow and stifle joints. The expression analysis showed 171 differentially expressed genes (DEGs), 113 were upregulated and 58 were downregulated compared to control dogs P (< 0.01). This pool of genes was functional annotated for signaling pathways using PANTHER tools. No overrepresented pathways were found. To gain further insights of the functional role of the DEGs in OA, we set a threshold of log2FoldChange value between -1.5 and 1.5. We ended up with 24 top up- and downregulated transcripts. Prioritization of these DEGs according to their known functional knowledge, revealed five possible candidates for OA biomarkers. The downregulated OSCAR gene encodes the osteoclast associated Ig-like receptor, which is involved in osteoclastogenesis regulation and bone homeostasis. In addition, the upregulated microRNA MIR339-1 and ncRNAs: LOC106559235 (downregulated), LOC102156762 (downregulated) and LOC111096460 (upregulated) are regulatory sequences, stable for gene profiling assessment in blood and related to OA pathogenesis regulation. We suggest OSCAR as the more likely candidate biomarker for OA diagnosis in dogs and, provide evidence of new circulating regulatory sequences differentially expressed in canine OA.

Objective The aim of this study was to investigate the prevalence of cranial cruciate ligament rupture and patellar luxation and the associated risk factors in dogs. Materials and Methods A total of 13,072 clinical records of dogs were reviewed from School Hospital (Faculty of Veterinary Sciences, National University of La Plata). Data of age, breed, sex, body weight, patellar luxation and cranial cruciate ligament rupture condition were registered. Chi-squared and Fisher's exact tests were used to compare the prevalence of cranial cruciate ligament rupture and patellar luxation with the variables and then univariable logistic regression was used to evaluate the risk of having cranial cruciate ligament rupture and patellar luxation. Multivariable logistic regression was used including all variables to assess the odds of having patellar luxation and cranial cruciate ligament rupture. Results Of 13,072 patients treated, 72 and 51 had cranial cruciate ligament rupture and patellar luxation respectively. Sex was not a major risk factor for either condition. Adult (odds ratio [OR] = 8.2) and senior (OR = 4.3) patients had increased risk of having cranial cruciate ligament rupture, while for patellar luxation age was not a risk factor. Groups 2, 3 and 8 were more likely to have cranial cruciate ligament rupture (OR = 5.5, OR = 9.1 and OR = 2.6), and group 11 had lower risk of having patellar luxation (OR = 0.08). Maxi (OR = 2.4) and giant (OR = 6.0) breeds had higher risk of having cranial cruciate ligament rupture, and medium and maxi breeds had higher risk of patellar luxation (OR = 0.05 and OR = 0.3). Multivariate OR test confirmed that age (adult), body size (giant and maxi) and breed group (Group 3) were significantly associated with having cranial cruciate ligament rupture, and age was associated with having patellar luxation. Clinical Significance This is the first epidemiological study of cranial cruciate ligament rupture and patellar luxation in dogs from School Hospital (Faculty of Veterinary Sciences, National University of La Plata). Giant and large adult dogs from the Molossoid and Terrier breeds were more likely to have cranial cruciate ligament rupture, while mixed and large dog breeds showed the lowest risk of having patellar luxation.

Glutathione (GSH) is an antioxidant synthesized from three constitutive amino acids (CAA): cysteine (Cys), glycine (Gly) and glutamate (Glu). Glutathione plays an important role in oocyte maturation, fertilization andearly embryo development. This study aimed to investigate the effect of Cys (0.6 mM), Gly (0.6 mM) and Glu (0.9 mM) supplementation during in vitro fertilization (IVF) of cattle oocytes. In a Pilot Experiment, de novo synthesis of GSH in bovine zygote was evaluated using a modified TALP medium prepared without MEM-essential and MEM-non-essential amino acids (mTALP): mTALP+CAA (constitutive amino acids); mTALP+CAA+5 mMBSO (buthionine sulfoximide); mTALP+Cys+Gly; mTALP+Cys+Glu and mTALP+Gly+Glu. This evidence led us to investigate the impact of CAA supplementation to TALP medium (with essential and non-essential amino acids) on zygote viability, lipid peroxidation, total intracellular GSH content (include reduced and oxidized form; GSH-GSSG), pronuclear formation in zygotes and subsequent embryo development. IVF media contained a) TALP; b) ALP+Cys+Gly+Glu (TALP+CAA); c) TALP+Cys+Gly; d) TALP+Cys+Glu; e) TALP+Gly+Glu, were used. Total GSH-GSSG concentration was increased in TALP, TALP+CAA, and TALP+Cys+Gly. The viability of zygote was similar among treatments. Lipid peroxidation was increased in zygote fertilized with TALP+Cys+Gly; TALP+Cys+Glu;TALP+Gly+Glu and TALP+CAA. The percentage of penetrated oocytes decreased in TALP+CAA and TALP+Cys+Gly. The cleavage rate was lower in TALP+CAA and TALP+Gly+Glu. The percentage of embryos developing to the blastocyst stage was lower in TALP+Cys+Glu and TALP+CAA. In conclusion, we have demonstrated the synthesis of GSH during IVF. However, Cys, Gly and Glu supplementation to TALPmedium had negative effects on embryonic development. Fil: Anchordoquy, Juan Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Ciencias Básicas. Cátedra de Fisiología; Argentina Fil: Lizarraga, Raúl Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Anchordoquy, Juan Mateo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Ciencias Básicas. Cátedra de Fisiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Nikoloff, Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Rosa, Diana Esther. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Ciencias Básicas. Cátedra de Fisiología; Argentina Fil: Fabra, Mariana Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Furnus, Cecilia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina

Debido al Aislamiento Social, Obligatorio y Preventivo (ASPO) por el COVID-19, la Universidad Nacional de La Plata comunicó a sus docentes la necesidad de mantener la continuidad pedagógica en todas sus dependencias. En la Facultad de Ciencias Veterinarias las TIC´S se comenzaron a utilizar ampliamente en los últimos años para mejorar los modelos formativos y organizativos que sustentan el aprendizaje. El objetivo fue describir la experiencia en el desarrollo del curso Genética General durante el ASPO-COVID-19, mediado por TIC´S y analizar el desempeño de las cohortes 2019 (cursada presencial) y 2020 (cursada virtual). Se utilizó el entorno virtual Moodle 3.1 como soporte didáctico. Las Actividades Presenciales Obligatorias se convirtieron en Actividades Virtuales Obligatorias. El total de alumnos en estudio fue de 296 de los cuales 150 corresponden a la cursada del 2019 y 146 a la del primer cuatrimestre 2020. Cuando se compararon las condiciones de cada alumno, no se encontraron diferencias significativas entre cantidad de promocionados, aprobados, desaprobados y ausentes entre los dos grupos. El uso de las TIC´S durante el ASPO nos permitió desarrollar el curso sin mayores inconvenientes; fortaleciendo la relación entre docentes y estudiantes.

Evidence of cerebellar abiotrophy (CA) was found in a six-month-old Arabian filly with signs of incoordination, head tremor, wobbling, loss of balance and falling over, consistent with a cerebellar lesion. Normal hematology profile blood test and cerebrospinal fluid analysis excluded infectious encephalitis, and serological testing for Sarcocystis neurona was negative. The filly was euthanized. Postmortem X-ray radiography of the cervical cephalic region identified not abnormalities, discounting spinal trauma. The histopathological analysis of serial transverse cerebellar sections by electron microscopy revealed morphological characteristics of apoptotic cells with pyknotic nuclei and degenerate mitochondria, cytoplasmic condensation and areas with absence of Purkinje cells, matching with CA histopathological characteristics. The indirect DNA test for CA was positive in the filly, and DNA test confirmed the CA carrier state in the parents and the recessive inheritance of the disease. To our knowledge this is the first report of a CA case in Argentina., Instituto de Genética Veterinaria, Facultad de Ciencias Veterinarias

Chia, sesame and flax seeds are becoming increasingly more frequent components of functional foods because of their proven health benefits. The identification of plant components in complex matrices is generally performed using DNA typing. This method is a fast and economic tool widely applied to assess the genetic origin and authenticity of products from the food supply chain. The aim of this study was to compare two DNA-based methods, quantitative polymerase chain reaction high-resolution melting (qPCR-HRM) and endpoint PCR, used to identify chia, flax and sesame seeds in laboratory-made and commercial products. For this purpose, an endpoint PCR and qPCR-HRM methods were developed. DNA was extracted from chia, sesame and flax seeds, laboratory-made and commercial products. The analysis of flax, chia and sesame melting profiles using HRM post-PCR analysis allowed the identification of chia, sesame and flax in processed foods. The results obtained showed that qPCR-HRM is a cost-effective and efficient method for the identification, authentication and/or detection of seeds in processed food and complex matrices containing chia, sesame and flax.

The Argentine Polo Horse (AP), an autochthonous breed officially created in Argentina in the early 1980s, is globally recognized as the best equid for playing polo. Their breeding is characterized by the use of cutting-edge assisted reproductive techniques such as large-scale embryo transfer (ET) programs and cloning. The aim of this study was to determine the impact of the use of those reproductive biotechnologies on the genetic structure, variability and reproductive parameters of the AP breed using genealogical data (81,633 pedigree records). In total, 18,077 animals drawn from the last generation (2006–2015) were employed as the reference population (WP), which was further divided into two subsets: animals produced (ET; n = 13,478) and not produced by ET (NOT-ET; n = 4,599). Horses produced by ET showed a significant decrease in generation interval compared with NOT-ET. On the other hand, the number of foals per stallion and broodmare as well as inbreeding (F = 0.89%) and average relatedness (AR=1%) were higher in ET compared with NOT-ET (F = 0.6%; AR = 0.54%), depicting an increased selective intensity. Our analysis also revealed that the effective number of founders and ancestors in ET showed a disproportionate gene contribution and a strong genetic bottleneck as well as the inter-herd fixation index (FST) revealed an increased genetic flow among herds and higher internal relatedness values within ET horses. In conclusion, the use of large-scale ET programs decreased genetic variability (lower effective population size and number of founders and ancestors and higher F, AR and coancestry), increased the genetic flow among herds and decreased the generation interval, contributing to the possibility of higher rates of genetic progress in the AP.

Copy number variation (CNV) has been proved to be widespread in human, animal and plant genomes. Together with single nucleotide polymorphisms (SNPs), CVNs play a key role in genetic diversity. In this study, genome-wide detection of CNVs was performed based on SNP data from 24 Criollo Argentino horses genotyped with the GGP Equine70k array. Overall, 165 CNVs meeting stringent quality control criteria were identified and then aggregated into 87 CNV regions (CNVRs), representing a horse genome coverage of 13.69 Mb. Functional analysis of CNVRs allowed the identification of 337 genes implicated in a wide range of biological functions such as signal transducer activity (olfactory receptors), receptor activities and binding. Furthermore, enrichment analysis showed that the most represented protein classes (over 25%) were immunoglobulin receptor subfamily, immunoglobulins and major histocompatibility complex antigen (beta-2-microglobulin). To the best of our knowledge, this is the first report of CNV in Criollo Argentino horses. Fil: Corbi Botto, Claudia Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Morales Durand, Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Zappa, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Sadaba, Sebastian Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina

El gen ABCB1 codifica para la glicoproteínaP (Pgp), una proteína de membrana que transporta múltiples fármacos fuera de la célula. Este gen se expresa principalmente en la barrera hematoencefálica, cumpliendo también importantes funciones en otros órganos. En caninos, se ha informado una deleción de 4 pb en el exón 4, que genera una terminación prematura y una proteína no funcional. Los animales homocigotos para la mutación presentan neurotoxicidad al administrarles drogas como las avermectinas. Esta mutación ha sido comunicada principalmente en collie, border collie y otras razas de perros pastores. El objetivo del trabajo consistió en desarrollar un método de diagnóstico rápido de la mutación nt230 [del4] del gen ABCB1 basado en pirosecuenciación y validarla en una población local. Se analizaron 72 perros mediante pirosecuenciación y secuenciación directa. Los resultados obtenidos con ambas técnicas evidenciaron 100 % de concordancia. El cálculo de la frecuencia génica del alelo nt230 [del4] (q) mostró diferencias entre las razas tipificadas: collie (n=30 y q=0,32), border collie (n=17 y q=0,06), pastor de Shetland (n=12 y q=0) y otras (n=16 y q=0). El valor promedio obtenido de frecuencia génica del alelo nt230 [del4] resultó menor al reportado en otros países. En conclusión, el método desarrollado podría ser utilizado para la detección temprana de la mutación nt230 [del4] causal de la deficiencia de Pgp, siendo una herramienta muy útil para la reproducción controlada y la prevención de la neurotoxicidad en los pacientes frente a tratamientos con avermectinas y otras drogas sustrato de la Pgp., ABCB1 gene codes for Pglycoprotein (Pgp), a membrane protein that transports multiple drugs out of the cell. It is mainly expressed in the bloodbrain barrier, although it also plays a role in other organs. In dogs, a 4 bp deletion in exon 4 of ABCB1 gene has been reported. This mutation results in a premature stop codon and a nonfunctional protein. Homozygous animals for the deletion present neurotoxicity when administered drugs such as avermectins. This mutation is found mainly in collie, border collie and other related dog breeds. The aim of the present work was to develop a pyrosequencing method for rapid diagnosis of the ABCB1 gene nt230 [del4] mutation and validate it by direct sequencing in a local population. A total of 72 dogs were analyzed, obtaining a 100 % agreement between both methods. The genetic variation of the mutated allele (q) showed differences between the evaluated breeds: collie (n = 30; q = 0.32), border collie (n = 17; q = 0.06), Shetland sheep dog (n=12; q= 0), others (n=13; q = 0). On average, the q value was lower than reported in other countries. Pyrosequencing could be used for early diagnosis of Pgp deficiency, being a very useful tool for controlled reproduction in kennels and for the prevention of neurotoxicity in patients treated with avermectins and other Pgp substrate drugs., Facultad de Ciencias Veterinarias

The objective of this research was to investigate the development of a reliable fingerprint from elemental and isotopic signatures of Argentinean honey to assess its geographical provenance. Honey, soil, and water from three regions (Córdoba, Buenos Aires, and Entre Rı́os) were collected. The multielemental composition was determined by ICP-MS. δ13C was measured by isotopic ratio mass spectrometry, whereas the 87Sr/86Sr ratio was determined using thermal ionization mass spectrometry. The data were analyzed by chemometrics looking for the association between the elements, stable isotopes, and honey samples from the three studied areas. Honey samples were differentiated by classification trees and discriminant analysis using a combination of eight key variables (Rb, K/Rb, B, U, 87Sr/86Sr, Na, La, and Zn) presenting differences among the studied regions. The application of canonical correlation analysis and generalized procrustes analysis showed 91.5% consensus between soil, water, and honey samples, in addition to clear differences between studied areas. To the authors’ knowledge, this is the first report demonstrating the correspondence between soil, water, and honey samples using different statistical methods, showing that elemental and isotopic honey compositions are related to soil and water characteristics of the site of origin. Fil: Baroni, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina Fil: Podio, Natalia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina Fil: Badini, Raúl G.. Provincia de Córdoba. Ministerio de Ciencia y Técnica. Centro de Excelencia en Productos y Procesos de Córdoba; Argentina Fil: Inga, Marcela. Provincia de Córdoba. Ministerio de Ciencia y Técnica. Centro de Excelencia en Productos y Procesos de Córdoba; Argentina Fil: Ostera, Hector Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geocronología y Geología Isotopica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geocronología y Geología Isotópica; Argentina Fil: Cagnoni, Mariana Celina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geocronología y Geología Isotopica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geocronología y Geología Isotópica; Argentina Fil: Gautier, Eduardo Amilcar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Hoogewerff, Jurian. University Of Otago; Canadá Fil: Wunderlin, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina

The Brangus breed was developed to combine the superior characteristics of both of its founder breeds, Angus and Brahman. It combines the high adaptability to tropical and subtropical environments, disease resistance, and overall hardiness of Zebu cattle with the reproductive potential and carcass quality of Angus. It is known that the major histocompatibility complex (MHC, also known as bovine leucocyte antigen: BoLA), located on chromosome 23, encodes several genes involved in the adaptive immune response and may be responsible for adaptation to harsh environments. The objective of this work was to evaluate whether the local breed ancestry percentages in the BoLA locus of a Brangus population diverged from the estimated genome-wide proportions and to identify signatures of positive selection in this genomic region. For this, 167 animals (100 Brangus, 45 Angus and 22 Brahman) were genotyped using a high-density single nucleotide polymorphism array. The local ancestry analysis showed that more than half of the haplotypes (55.0%) shared a Brahman origin. This value was significantly different from the global genome-wide proportion estimated by cluster analysis (34.7% Brahman), and the proportion expected by pedigree (37.5% Brahman). The analysis of selection signatures by genetic differentiation (F st ) and extended haplotype homozygosity-based methods (iHS and Rsb) revealed 10 and seven candidate regions, respectively. The analysis of the genes located within these candidate regions showed mainly genes involved in immune response-related pathway, while other genes and pathways were also observed (cell surface signalling pathways, membrane proteins and ion-binding proteins). Our results suggest that the BoLA region of Brangus cattle may have been enriched with Brahman haplotypes as a consequence of selection processes to promote adaptation to subtropical environments. Fil: Goszczynski, Daniel Estanislao. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Corbi Botto, Claudia Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Durand, H. M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Rogberg Muñoz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Unidad Ejecutora de Investigaciones en Producción Animal. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Unidad Ejecutora de Investigaciones en Producción Animal; Argentina Fil: Munilla Leguizamon, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Unidad Ejecutora de Investigaciones en Producción Animal. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Unidad Ejecutora de Investigaciones en Producción Animal; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Cantet, Rodolfo Juan Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Unidad Ejecutora de Investigaciones en Producción Animal. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Unidad Ejecutora de Investigaciones en Producción Animal; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina

Adequate dietary intake of copper (Cu) is required for normal reproductive performance in cattle. The objective of this study was to investigate the pregnancy rates from cattle with deficient, marginal and adequate Cu plasma concentration at the beginning of artificial insemination protocol. Moreover, we determined Cu concentrations present in bovine oviductal fluid (OF), and the effects of Cu on fertilizing ability of bovine spermatozoa. Also, the presence of Cu transporter, SLC31A1 (also known as CTR1), in spermatozoa and in vitro matured oocyte were investigated. We found no differences in pregnancy rates among animals with adequate, marginal, and deficient Cu concentrations measured in plasma at the beginning of fixed-time artificial insemination (FTAI) protocol. Copper concentrations in OF were 38.3 ± 2.17 μg/dL (mean ± SEM) regardless of cupremia levels. The addition of 40 μg/dL Cu to IVF medium enhanced total and progressive motility, sperm viability, functional sperm membrane integrity (HOST), sperm–zona binding, and pronuclear formation. On the other hand, the presence of Cu in IVF medium did not modify acrosome integrity and cleavage rates after IVF, but impaired blastocyst rates. Cu transporter SLC31A1 was detected in bovine spermatozoa in the apical segment of acrosome, and in the oocyte matured in vitro. In conclusion, the results obtained in the present study determined that cupremia levels at the beginning of FTAI protocol did not influence the pregnancy rates at 60 d after insemination. The presence of CTR1 in bovine mature oocyte and spermatozoa, as well as the beneficial effect of Cu on sperm quality would suggest an important role of this mineral during the fertilization process. Fil: Anchordoquy, Juan Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Anchordoquy, Juan Mateo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Pascua, Ana Malen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Nikoloff, Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Furnus, Cecilia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina

In bovine, there are significant differences among and within breeds in the time when bulls reach puberty. Reported data indicate that age at puberty has a strong genetic component and is a multigenic trait. However, despite significant advancements in the field, the gene network controlling puberty is very complex, containing a host of genes that exert their effect on the hypothalamic–pituitary–gonadal (HPG) axis, and several aspects of regulation of the HPG axis are yet to be fully elucidated. The objective of this work was to identify new genes associated with age at puberty. Angus males (N=276) were weighted and scrotal circumference measured every month. When first bull calves reached 26 cm of scrotal circumference, sperm quality was added to the monthly measurements for the next three months. Based on sperm quality, two groups corresponding to the top and bottom 6.5% of the phenotypic distribution curve were selected to perform a genome-wide association study using a selective DNA approach, to be used as an exploratory analysis. Based on the results of this preliminary study, five SNPs located within BTA3, 20 and 24 associated regions were genotyped in the 276 bulls using pyrosequencing methods. This analysis allowed us to confirm the association of a BTA20 region with age at puberty estimated at C 50 million. The associated SNP explained 1.69% of the genetic variance of age at puberty at C 50 million. The associated region contained within and nearby positional candidate genes (ISL1, PELO, FST, SPZ1) that may be involved in spermatogenesis. Our results should encourage further investigation to confirm the role of these genes in order to increase our knowledge on genetic control of bovine puberty. Fil: Fernandez, María Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Liron, Juan Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Prando, Alberto. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Rogberg Muñoz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Baldo A. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Giovambattista, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina

The aim of this study was to investigate the influence of zinc (Zn) on the health of cumulus-oocyte complex (COC) during in vitro maturation (IVM). Experiments were designed to evaluate the effect of Zn added to IVM medium on: DNA integrity, apoptosis, cumulus expansion and superoxide dismutase (SOD) activity of cumulus cells (CC). Also, role of CC on Zn transport during IVM was evaluated on oocyte developmental capacity. DNA damage and early apoptosis were higher in CC matured with 0 μg/ml Zn compared with 0.7, 1.1 and 1.5 μg/ml Zn (p 0.05). Cumulus expansion did not show differences in COC matured with or without Zn supplementation (p 0.05). Superoxide dismutase activity was higher in COC matured with 1.5 μg/ml Zn than with 0 μg/ml Zn (p 0.05). Cleavage and blastocyst rates were recorded after IVM in three maturation systems: intact COCs, denuded oocytes with cumulus cells monolayer (DO + CC) and denuded oocytes (DO). Cleavage rates were similar when COC, DO + CC or DO were matured with 1.5 μg/ml Zn compared with control group (p 0.05). Blastocyst rates were significantly higher in COC than in DO + CC and DO with the addition of 1.5 μg/ml Zn during IVM (p 0.01). Blastocyst quality was enhanced in COC and DO + CC compared with DO when Zn was added to IVM medium (p 0.001). The results of this study indicate that Zn supplementation to IVM medium (i) decreased DNA damage and apoptosis in CC; (ii) increased SOD activity in CC; (iii) did not modify cumulus expansion and cleavage rates after in vitro fertilization; (iv) improved subsequent embryo development up to blastocyst stage; and (v) enhanced blastocyst quality when CC were present either in intact COC or in coculture during IVM.

Chromosomal abnormalities in the sex chromosome pair (ECAX and ECAY) are widely associated with reproductive problems in horses. However, a large proportion of these abnormalities remains undiagnosed due to the lack of an affordable diagnostic tool that allows for avoiding karyotyping tests. Hereby, we developed an STR (single-tandem-repeat)- based molecular method to determine the presence of the main sex chromosomal abnormalities in horses in a fast, cheap and reliable way. The frequency of five ECAXIinked (LEX026, LEX003, TKY38, TKY270 and UCDE0502) and two ECAY-Iinked (EcaYH12 and SRY) markers was characterized in 261 Purebred Spanish Horses to determine the efficiency of the methodology developed to be used as a chromosomal diagnostic tool. All the microsatellites analyzed were highly polymorphic, with a sizeable number of alleles (polymorphic information content > 0.5). Based on this variability, the methodology showed 100% sensitivity and 99.82% specificity to detect the most important sex chromosomal abnormalities reported in horses (chimerism, Turner’s syndrome and sex reversal syndromes). The method was also validated with 100% efficiency in 10 individuals previously diagnosed as chromosomally aberrant. This STR screening panel is an efficient and reliable molecular-cytogenetic tool for the early detection of sex chromosomal abnormalities in equines that could be included in breeding programs to save money, effort and time of veterinary practitioners and breeders., Facultad de Ciencias Veterinarias

The case of an identification for a small bone found in a blood sausage is described. The similitude with a rodent bone leads the consumer to demand the government organism responsible of food quality control. Bone and blood sausage DNA analysis was performed in order to determinate the origin species and the possibility of food contamination and/or adulteration. DNA sequence analysis confirmed the pig origin of both samples with an identity higher than 98%., Instituto de Genética Veterinaria

Escherichia coli O157:H7 is an enteric pathogen associated with food safety threats and with significant morbidity and mortality worldwide. In Argentina, post-enteric hemolytic uremic syndrome (HUS) is endemic, with >70% of cases associated with E. coli O157 infection. To date the biological basis behind the severity among E. coli O157 infections is unknown. However, single nucleotide polymorphism (SNP) typing has helped to define nine E. coli O157:H7 clades, of which clade 8 strains are associated with severe disease cases. The aim of this study was to characterize a collection of 20 STEC O157:H7 strains isolated between 2011 and 2013 from ground beef and different environmental samples from butcher shops of Argentina. All strains harbored the eae, ehxA, fliCH7, efa, iha, and toxB genes, with stx2a/stx2c as the predominant genotype (75%). The XbaI-PFGE analysis showed that the E. coli O157 strains had high genetic diversity. Nine strains were grouped in four XbaI-PFGE clusters, whereas 11 strains showed unique XbaI-PFGE patterns. In contrast, the SNP analysis allowed us to separate the strains in two distinct lineages representing clade 8 (70%) and clade 6 (30%). Our results show the molecular characterization of E. coli O157 strains isolated from ground beef and environmental samples from Argentinean butcher shops., Instituto de Genética Veterinaria

Canine chronic superficial keratitis (CSK) is an inflammatory corneal disease that primarily occurs in German shepherd dogs (GSDs). Several studies support the hypothesis that CSK is an immune-mediated disease. To investigate the genetic factors associated with CSK development, the upstream regulatory regions (URRs) of the DLA-DRB, -DQA and -DQB genes were genotyped in 60 dogs, including 32 CSK animals. LD analysis identified two blocks (r2 ≤ 45), with two DLA-DRB1 and five DLA-DQB1 haplotypes. Analysis of DLA-URR alleles/haplotypes showed a significant association between DQB1*-154 [C/T] (p = 0.016) and CSK, suggesting that the T variant may increase the risk for developing CSK disease (OR = 3, 95% CI = 1.25–7.68). When haplotype associations were performed, the URR-DQB*CATT haplotype was significantly associated with CSK (p = 0.016), increasing the risk of develop this disease over two-fold (OR = 3, 95%, CI = 1.25–7.68). These results showed that dogs homozygous at DRB1*69 [C/T] had a risk for developing CSK disease that was over four times the risk for heterozygotes. This genetic association supports the previous clinical, histological and pharmacological studies that suggest that CSK is an immune-mediated disease, and this association could potentially be used to identify susceptible animals. Fil: Barrientos, Laura Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Zapata, Gustavo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Crespi, Julian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Posik, Diego Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: It, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina

Equine lentivirus receptor-1 (ELR1) has been characterized as the specific functional receptor that mediates equine infectious anemia virus (EIAV) entrance to horse macrophages. This receptor is tumor necrosis factor receptor superfamily member 14 (TNFRSF14). The aim of this study was to investigate the occurrence of allelic variants in the coding sequence of equine TNFRSF14 gene by screening for single-nucleotide polymorphisms (SNPs) in different equine populations. Forty seven horse samples were randomly selected from a reservoir of EIAV-seropositive and seronegative samples collected from different outbreaks and regions of Argentina. DNA samples were scanned via PCR and direct sequencing of exon 3 and exon 5 of TNFRSF14 gene. A total of 21 SNPs were identified, of which 11 were located in coding sequences. Within exon 5, four SNPs caused nonsynonymous substitutions, while two other SNPs caused synonymous substitutions in crucial residues (Ser112 and Thr114) implicated in the interaction with EIAV. Despite some of exon 5 variants occurred exclusively in EIAV-positive or EIAV-negative horses, critical residues for the function of the mature protein were conserved, accounting for selective pressures in favor of preserving the specific function of TNFRSF members and the host immune response. To our knowledge, this is the first report of the existence of allelic variations involving some crucial amino acid residues in horse ELR1. Further, it could be an initial step to test the possible functional relevance and relationship of these variants with EIAV infection and disease progression as well as to develop preventive strategies. Fil: Corbi Botto, Claudia Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Sadaba, Sebastian Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Zappa, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina

Dog fecal samples were collected at the crime scene and from the shoes of the suspectto see whether they could be linked. DNA was genotyped using a 145 bp fragmentcontaining a 60 bp hotspot region of the mitochondrial DNA (mtDNA) control region.Once the species origin was identified, sequences were aligned with the 23 caninehaplotypes defined, showing that evidence and reference had 100% identity withhaplotype 5. The frequency of haplotype 5 and the exclusion power of the referencepopulation were 0.056 and 0.89, respectively. The forensic index showed that it was 20times more likely that the evidence belonged to the reference dog than to some otherunknown animal. The results support that the mtDNA hypervariable region 1 (HV1) isa good alternative for typing in trace or degraded casework samples when the STRpanel fails, and demonstrate the utility of domestic animal samples to give additionalinformation to solve human legal cases. Fil: Barrientos, Laura Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Crespi, Julian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Fameli, Alberto Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina Fil: Posik, Diego Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Morales, Hernan Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina

Adequate dietary intake of manganese (Mn) is required for normal reproductive performance in cattle. This study was carried out to investigate the effect of Mn during in vitro maturation of bovine cumulus–oocyte complexes (COC) on apoptosis of cumulus cells, cumulus expansion, and superoxide dismutase (SOD) activity in the COC. The role of cumulus cells on Mn transport and subsequent embryo development was also evaluated. Early apoptosis decreased in cumulus cells matured with Mn compared with medium alone. Cumulus expansion did not show differences in COC matured with or without Mn supplementation. SOD activity was higher in COC matured with 6 ng/ml Mn than with 0 ng/ml Mn. Cleavage rates were higher in COC and denuded oocytes co-cultured with cumulus cells, either with or without Mn added to in vitro maturation (IVM) medium. Regardless of the presence of cumulus cells during IVM, the blastocyst rates were higher when 6 ng/ml Mn was supplemented into IVM medium compared with growth in medium alone. Blastocyst quality was enhanced when COC were matured in medium with Mn supplementation. The results of the present study indicated that Mn supplementation to IVM medium enhanced the ‘health’ of COC, and improved subsequent embryo development and embryo quality. Fil: Anchordoquy, Juan Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Laboratorio de Nutricion Mineral y Fisiologia Reproductiva; Argentina Fil: Anchordoquy, Juan Mateo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Laboratorio de Nutricion Mineral y Fisiologia Reproductiva; Argentina Fil: Sirini, Matias Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Laboratorio de Nutricion Mineral y Fisiologia Reproductiva; Argentina Fil: Testa, Juan Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de la Plata. Facultad de Cs.veterinarias. Departamento de Cs.basicas. Laboratorio de Nutricion Mineral y Fisiologia Reproductiva; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Furnus, Cecilia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina

Foodborne pathogens can cause acute and chronic diseases and produce a wide range of symptoms. Since the consumption of ground beef is a risk factor for infections with some bacterial pathogens, we performed a comprehensive evaluation of butcher shops, implemented improvement actions for both butcher shops and consumers, and verified the impact of those actions implemented. A comprehensive evaluation was made and risk was quantified on a 1-100 scale as high-risk (1-40), moderate-risk (41-70) or low-risk (71-100). A total of 172 raw ground beef and 672 environmental samples were collected from 86 butcher shops during the evaluation (2010-2011) and verification (2013) stages of the study. Ground beef samples were analyzed for mesophilic aerobic organisms, Escherichia coli and coagulase-positive Staphylococcus aureus enumeration. Salmonella spp., E. coli O157:H7, non-O157 Shiga toxin-producing E. coli (STEC), and Listeria monocytogenes were detected and isolated from all samples. Risk quantification resulted in 43 (50.0%) high-risk, 34 (39.5%) moderate-risk, and nine (10.5%) low-risk butcher shops. Training sessions for 498 handlers and 4,506 consumers were held. Re-evaluation by risk quantification and microbiological analyses resulted in 19 (22.1%) high-risk, 42 (48.8%) moderate-risk and 25 (29.1%) low-risk butcher shops. The count of indicator microorganisms decreased with respect to the 2010-2011 period. After the implementation of improvement actions, the presence of L. monocytogenes, E. coli O157:H7 and stx genes in ground beef decreased. Salmonella spp. was isolated from 10 (11.6%) ground beef samples, without detecting statistically significant differences between both study periods (evaluation and verification). The percentage of pathogens in environmental samples was reduced in the verification period (Salmonella spp., 1.5%; L. monocytogenes, 10.7%; E. coli O157:H7, 0.6%; non-O157 STEC, 6.8%). Risk quantification was useful to identify those relevant facts in butcher shops. The reduction of contamination in ground beef and the environment was possible after training handlers based on the problems identified in their own butcher shops. Our results confirm the feasibility of implementing a comprehensive risk management program in butcher shops, and the importance of information campaigns targeting consumers. Further collaborative efforts would be necessary to improve foodstuffs safety at retail level and at home., Facultad de Ciencias Veterinarias, Instituto de Genética Veterinaria

The main goal of this study was to propose a reliable method to verify the geographical origin of meat, establishing the influence of soil and water on its isotopic and elemental composition. Thus, beef meat, soil, and water samples were collected from three major cattle-producing regions of Argentina (Buenos Aires, Córdoba, and Entre Ríos). Multielemental composition was determined on these three matrices by inductively coupled plasma mass spectrometry (ICP-MS), δ(13)C and δ(15)N by isotope-ratio mass spectrometry (IRMS), and the (87)Sr/(86)Sr ratio by thermal ionization mass spectrometry (TIMS). Soil and drinking water samples could be characterized and clearly differentiated by combining the isotopic ratios and elements, demonstrating differences in geology and climatic conditions of three regions. Similarly, meat originating at each sampling area was characterized and differentiated using only five key variables (Rb, Ca/Sr, δ(13)C, δ(15)N, and (87)Sr/(86)Sr). Generalized procrustes analysis (GPA), using the three studied matrices (soil, water, and meat) shows consensus between them and clear differences between studied areas. Furthermore, canonical correlation analysis (CCA) demonstrates significant correlation between the chemical-isotopic profile of meat with those corresponding to both soil and water (r(2) = 0.93, p0.001; and r(2) = 0.83, p0.001, respectively). So far, there are clear coincidences between the meat fingerprint and those from soil/water where cattle grew, presenting a good method to establish beef provenance. To the authors' knowledge this is the first report linking the influence of soil and water all together on the composition of beef, presenting the basis for the authentication of Argentinean beef, which could be extended to meat from different provenances.

Our main goal was to investigate if robust chemical fingerprints could be developed for three Argentinean red wines based on organic, inorganic, and isotopic patterns, in relation to the regional soil composition. Soils and wines from three regions (Mendoza, San Juan, and Córdoba) and three varieties (Cabernet Sauvignon, Malbec, and Syrah) were collected. The phenolic profile was determined by HPLC-MS/MS and multielemental composition by ICP-MS; (87)Sr/(86)Sr and δ(13)C were determined by TIMS and IRMS, respectively. Chemometrics allowed robust differentiation between regions, wine varieties, and the same variety from different regions. Among phenolic compounds, resveratrol concentration was the most useful marker for wine differentiation, whereas Mg, K/Rb, Ca/Sr, and (87)Sr/(86)Sr were the main inorganic and isotopic parameters selected. Generalized Procrustes analysis (GPA) using two studied matrices (wine and soil) shows consensus between them and clear differences between studied areas. Finally, we applied a canonical correlation analysis, demonstrating significant correlation (r = 0.99; p0.001) between soil and wine composition. To our knowledge this is the first report combining independent variables, constructing a fingerprint including elemental composition, isotopic, and polyphenol patterns to differentiate wines, matching part of this fingerprint with the soil provenance.

Genetic markers have been used to assess the association of economically important traits with cattle under intensive feeding conditions; however, there is still the need to ascertain the usefulness of these markers under extensive production systems. Bovine chromosome 5 has been widely studied because several QTLs have been detected. Microsatellite BP1 neighboring the Myogenic factor 5 gene (Myf5), and microsatellites ETH10, IGF1 and RM029 near Insulin-like Growth Factor 1 gene (IGF1), were selected to establish their association with BLUPs (Best Linear Unbiased Predictor) for direct Birth Weight (dBW), direct Weaning Weight (dWW) and maternal Weaning Weight (mWW). Two herds were used for this objective, one commercial and the other experimental. Associations (P ≤ 0.05) between dWW and all BTA5 loci (BP1, ETH10, IGF1, and RM029) were detected. Additional associations were observed between mWW and BP1. dBW was significantly associated (P ≤ 0.05) with ETH10 genotypes and with the interaction IGF1*Herd. In particular region near BP1 could be contributing to the rare positive correlation between dWW and mWW previously found in the INTA Balcarce Station experimental herd. We confirmed marker associations with growth traits in two BTA5 regions close to previously reported QTL obtained in intensive feeding conditions; these regions affect dBW, dWW and mWW in a pasture-based system. Fil: Rogberg Muñoz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Melucci, Lilia Magdalena. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina Fil: Prando, Alberto José. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Villegas Castagnasso, Egle Etel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Ripoli, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Baldo, Andres. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Añon, Maria Cristina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina

Glutathione (GSH) concentration increases in bovine oocytes during in vitro maturation (IVM). The constitutive amino acids involved in GSH synthesis are glycine (Gly), glutamate (Glu) and cysteine (Cys). The present study was conducted to investigate the effect of the availability of glucose, Cys, Gly and Glu on GSH synthesis during IVM. The effect of the amino acid serine (Ser) on intracellular reduced/oxidized glutathione (GSH/GSSG) content in both oocytes and cumulus cells was also studied. Cumulus-oocyte complexes (COC) of cattle obtained from ovaries collected from an abattoir were matured in synthetic oviduct fluid (SOF) medium containing 8 mg/ml bovine serum albumin-fatty acid-free (BSA-FAF), 10 microg/ml LH, 1 microg/ml porcine FSH (pFSH) and 1 microg/ml 17 beta-estradiol (17beta-E2). GSH/GSSG content was measured using a double-beam spectrophotometer. The COC were cultured in SOF supplemented with 1.5mM or 5.6mM glucose (Exp. 1); with or without Cys+Glu+Gly (Exp. 2); with the omission of one constitutive GSH amino acid (Exp. 3); with 0.6mM Cys or Cys+Ser (Exp. 4). The developmental capacity of oocytes matured in IVM medium supplemented with Cys and the cell number per blastocyst were determined (Exp. 5). The results reported here indicate (1) no differences in the intracellular GSH/GSSG content at any glucose concentrations. Also, cumulus cell number per COC did not differ either before or after IVM (Exp. 1). (2) Glutathione content in oocytes matured in SOF alone were significantly different from oocytes incubated with SOF supplemented with Cys+Glu+Gly (Exp. 2). (3) Addition of Cys to maturation medium, either with or without Gly and Glu supplementation resulted in an increase of GSH/GSSG content. However, when Cys was omitted from the IVM medium intracellular GSH in oocytes or cumulus cells was less but not significantly altered compared to SOF alone (Exp. 3). (4) Glutathione content in both oocytes and cumulus cells was significantly reduced by incubation with 5mM Ser (Exp.4). (5) There was a significant increase in cleavage and blastocyst rates when Cys was added to maturation medium. In contrast, the cleavage, morula and blastocyst rates were significantly different when 5mM Ser was added to maturation media. There was also a significant difference in mean cell number per blastocyst, obtained from oocytes matured with 5mM Ser (Exp. 5). This study provides evidence that optimal embryo development in vitro is partially dependent on the presence of precursor amino acids for intracellular GSH production. Moreover, the availability of Cys might be a critical factor for GSH synthesis during IVM in cattle oocytes. Greater Ser concentration in IVM medium altered "normal" intracellular GSH in both oocytes and cumulus cells with negative consequences for subsequent developmental capacity.

Breed assignment has proved to be useful to control meat trade and protect the value of special productions. Meat-related frauds have been detected in China; therefore, 95 SNPs selected from the ISAG core panel were evaluated to develop an automated and technologically updated tool to screen breed label fraud in the Chinese meat market. A total of 271 animals from four Chinese yellow cattle (CYC) populations, six Bos taurus breeds, two Bos indicus and one composite were used. The allocation test distinguished European, Japanese and Zebu breeds, and two Chinese genetic components. It correctly allocated Japanese Black, Zebu and British breeds in 100, 90 and 89% of samples, respectively. CYC evidenced the Zebu, Holstein and Limousin introgression. The test did not detect CYC components in any of the 25 samples from Argentinean butchers. The method could be useful to certify Angus, Hereford and Japanese Black meat, but a modification in the panel would be needed to differentiate other breeds. Fil: Rogberg Muñoz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Wei, S.. Chinese Academy of Agricultural Sciences; China Fil: Ripoli, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Guo, B. L.. Chinese Academy of Agricultural Sciences; China Fil: Carino, Mónica Herminia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Liron, Juan Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Prando, Alberto José. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Vaca, Roberto Jose Antonio. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina Fil: Wei, Y. M.. Chinese Academy of Agricultural Sciences; China Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria ; Argentina

El presente libro nace como producto de una convocatoria para Libros de Cátedra de la Universidad Nacional de la Plata. La propuesta, avalada por la Facultad de Ciencias Veterinarias, permitirá introducir al lector en los principales aspectos de la identificación genética de animales y/o sus productos derivados, conocer los principales métodos de genotipificación utilizados en genética forense, reconocer los lineamientos de estandarización y acreditación de los laboratorios de genética forense y por ultimo fortalecer las capacidades de los futuros profesionales en el área de genética. Entre los autores de esta obra se destacan profesionales de distintas entidades académicas: Universidades Nacionales (UNCPBA, UNLP), del exterior (Universidade Federal de Minas Gerais. Belo Horizonte, Brasil; Universidad de Western Cape Sudáfrica), laboratorios de servicios nacionales (Laboratorio de ADN, Facultad de Ciencias Veterinarias, UNCPBA, Tandil, de la CICPBA y el CONICET); internacionales (Laboratório de Genética, Escola de Veterinária y el Myleus Biotechnology Research Team, Belo Horizonte, Brasil) y profesionales del Cuerpo de Instructores de la Fiscalía General del Departamento Judicial Azul y de Junín de la Provincia de Buenos Aires. Los coordinadores del presente libro son profesionales responsables del dictado en la carrera de grado de Médico Veterinario de la UNLP, de un curso optativo de Genética Veterinaria Forense de la UNLP, y miembros del Servicio Externo de Diagnóstico Genético en Animales Domésticos del Instituto de Genética Veterinaria (IGEVET, UNLP-CONICET)., Facultad de Ciencias Veterinarias

In order to clarify the historical origin and phylogeographic affinities of Creole cattle matrilineages throughout the American continent, we analysed published D-loop mtDNA sequences (n = 454) from Creole, Iberian and African cattle breeds. The Western European T3 haplogroup was the most common in American Creole cattle (63.6%), followed by the African T1 (32.4%) and the Near Eastern T2 haplogroups (4%). None of the sequences were found in Bos indicus types. Within the African T1 haplogroup there were two subclades, T1a and T1*, whose geographic distribution in America was clearly disjointed. T1a is a highly divergent clade originally reported for Creole cattle from Brazil and the Lesser Antilles, but whose geographic distribution in Africa remains unknown. In contrast, lineages attributable to T1* are restricted in America to the region colonized by the Spaniards. We propose a new hypothesis for the origins of Creole cattle that summarizes all previously published historical and genetic data. While the African T1* fraction in Creole cattle may have arrived in America through the Iberian breeds, the divergent T1a lineages may have been introduced by Portuguese and other European crowns from some unknown, not-yet-sampled African location. Additional molecular studies will be required for pinpointing the specific African regional source. Fil: Liron, Juan Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina Fil: Bravi, Claudio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; Argentina Fil: Mirol, Patricia Monica. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Peral Garcia, Pilar. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Giovambattista, Guillermo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Four PCR-multiplexes of mini-STR loci have been used to get a nuclear DNA profile from different severely degraded casework biological specimens that generated negative PCR results or partial profiles when commercial STR kits were employed. Mini-STR technology allowed to retrieve additional genetic information with very high efficiency especially for those STR loci with allele sizes less than 150 bp. Our data indicate that the mini-STR technology is an effective strategy to improve DNA profiling from severely degraded casework human DNA samples that are refractory to amplification of DNA fragments bigger than 200–300 bp.

Age at puberty is an important component of reproductive performance in cattle, so it is important to identify genes that contribute to the regulation of the onset of puberty and polymorphisms that explain differences between bulls. In a previous study, we found putative associations between age at puberty in Angus bulls and single-nucleotide polymorphisms (SNPs) in Chromosomes 1 and X. In the present work we aimed to confirm these findings in a larger 5 sample of Angus bulls (n ¼ 276). Four SNPs located in these regions were genotyped using SEQUENOM technology and the genotypes obtained were tested for association with age at puberty. The results showed that SNPs rs135953349 and rs110604205 on BTA1 were still significantly associated with age of puberty estimated at progressive sperm motility of 10% (P , 0.05). The association previously found on Chromosome X could not be confirmed. Analysis of the bovine genome revealed that the associated region (99.17–99.99 Mb) contained four predicted loci: myelodysplasia syndrome 10 1 (MDS1) and ecotropic virus integration site 1 (EVI1) complex locus (MECOM), eGF-like and EMI domain-containing 1 pseudogene-like (LOC100337483), microRNA mir-551b (MIR551B) and mCG140927-like (LOC100139843). The results obtained could contribute to the understanding of puberty regulation and could be useful for further identification and annotation of gene function in the context of reproduction. Fil: Fernandez, María Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Prando, Alberto. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Rogberg Muñoz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Baldo, Andrés. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Liron, Juan Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina

Methods for individual identification are usually employed for traceability, whereas breed identification is useful to detect commercial frauds. In this study, Chinese Yellow Cattle (CYC) samples plus data from six Bos taurus breeds, two Bos indicus breeds, and one composite breed were used to develop an allocation test based on 22 microsatellites. The test allowed discriminating all foreign breeds from the CYC, although some CYC individuals were wrongly allocated as Limousin or Holstein, probably due to the recent introduction of these breeds into China. In addition, CYC evidenced a previously reported Zebu cline (south–north) and a possible structure within the B. taurus component that should be confirmed. An independent test performed with meat samples of unknown breed origin from Argentina allocated 92% of them to either Angus, Hereford, or their crossbreed, but none was identified as CYC. We conclude that the test is a suitable tool to certify meat of foreign breed origin and to detect adulterations of CYC beef labeled as imported meat. Fil: Rogberg Muñoz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Wei, S.. Chinese Academy of Agricultural Science; China Fil: Ripoli, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Guo, B. L.. Chinese Academy of Agricultural Science; China Fil: Carino, M. H.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Castillo, Nadia Sabiela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Villegas Castagnasso, Egle Etel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Liron, Juan Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Morales Durand, H. F.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Melucci, L.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina Fil: Villarreal, E.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Wei, Y. M.. Chinese Academy of Agricultural Science; China Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina

Short tandem repeats (STR)s have been the eligible markers for forensic animal genetics, despite single-nucleotide polymorphisms (SNP)s became acceptable. The technology, the type, and amount of markers could limit the investigation in degraded forensic samples. The performance of a 32-SNP panel genotyped through OpenArraysTM (real-time PCR based) was evaluated to resolve cattle-specific forensic cases. DNA from different biological sources was used, including samples from an alleged instance of cattle rustling. SNPs and STRs performance and repeatability were compared. SNP call rate was variable among sample type (average = 80.18%), while forensic samples showed the lowest value (70.94%). The repeatability obtained (98.7%) supports the used technology. SNPs had better call rates than STRs in 12 of 20 casework samples, while forensic index values were similar for both panels. In conclusion, the 32-SNPs used are as informative as the standard bovine STR battery and hence are suitable to resolve cattle rustling investigations. Fil: Fernandez, María Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Rogberg Muñoz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Liron, Juan Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Goszczynski, Daniel Estanislao. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Ripoli, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Carino, Mónica H.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Instituto de Genética Veterinaria "ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina

Body temperature responses and the timing of abortions were evaluated in pregnant bitches with the anti-progestin aglepristone. Fifteen purebred and crossbred, 25-45 days pregnant, were included in this study and seven untreated bitches at the same stage of pregnancy served as controls. Treated bitches were administered two applications of aglepristone (10 mg/kg SC) 24 h apart for pregnancy termination. Pregnancy termination was confirmed by ultrasonographic assessment. Body temperature was rectally measured three times a day for 6 days beginning 24 h before treatment or pregnancy diagnosis in the treated and control bitches, respectively. Additionally, serum progesterone concentrations were assessed at time points during the study in the treated bitches. Pregnancy was terminated in 14 treated bitches in a mean+/-S.E.M. of 4.3+/-0.7 days after treatment. Control bitches remained pregnant. In the treated bitches, but not in the controls, body temperature significantly decreased 24 h after the beginning of the treatments (P0.01) and then gradually returned to pre-treatment values. Correlation between the day of mean minimum body temperature and the day of pregnancy termination was low (0.07;0.05). Progesterone did not show significant change throughout the study. Body temperature does not seem to be a suitable variable to clinically monitor the aborting effect of aglepristone. Decrease of body temperature after aglepristone treatment could represent further evidence of its hypothalamic effects.

Sequence information was obtained on the variation of the ELA-DRB upstream regulatory region (URR) after polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) cloning and sequencing of ≈ 220 bp upstream of the first exon of horse DRB genes. The sequence of the proximal URR of equine DRB is composed of highly conserved sequence motifs, showing the presence of the W, X, Y, CAAT and TATA conserved boxes of major histocompatibility complex (MHC) class II promoters. Five different polymorphic horse DRB promoter sequences were detected in five horse breeds. The results demonstrate the existence of polymorphism in the nucleotide sequences of the ELA-DRB URR, located in the functionally important conserved consensus sequences, the X2 box, the Y box and the TATA box, while conservation were observed in X1 and CAAT boxes. The nucleotide diversity among horse URRs was intermediate between that seen within human and mouse DRB promoters, suggesting the existence of another important source of variability in ELA-DRB genes. In addition, phylogenetic comparisons, identity analysis and sequence organization suggested that the reported sequences would correspond to an expressed ELA-DRB locus. However, further information about the functional significance of these promoter polymorphisms will probably be acquired through expression studies on the different sequences. Fil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina

We developed a simple, quick assay in order to discriminate forensic samples among human, and common domestic and livestock species of the Pampean region, Argentina. A mitochondrial cytochrome b fragment amplified with universal primers was separately digested with three restriction enzymes (AluI, HaeIII, and HinfI) and the resulting fragments were resolved through electrophoresis in polyacrylamide gels. This PCR-RFLP method allowed us to identify the target species and worked on degraded samples. The assay was successfully applied in livestock robbery cases in Argentine, and may be useful when attempting a first assessment as to the specific status of a forensic evidence. © 2004 Elsevier Ireland Ltd. All rights reserved. Fil: Bravi, Claudio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina Fil: Liron, Juan Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina Fil: Mirol, Patricia Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. University of Leeds. School of Biology; Reino Unido Fil: Ripoli, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Centro de Investigaciones en Genética Básica y Aplicada; Argentina

We explore different designs to estimate both nuclear and mitochondrial human DNA (mtDNA) content based on the detection of the 5′ nuclease activity of the Taq DNA polymerase using fluorogenic probes and a real-time quantitative PCR detection system. Human mtDNA quantification was accomplished by monitoring the real-time progress of the PCR-amplification of two different fragment sizes (113 and 287 bp) within the hypervariable region I (HV1) of the mtDNA control region, using two fluorogenic probes to specifically determine the mtDNA copy of each fragment size category. This mtDNA real-time PCR design has been used to assess the mtDNA preservation (copy number and degradation state) of DNA samples retrieved from 500 to 1500 years old human remains that showed low copy number and highly degraded mtDNA. The quantification of nuclear DNA was achieved by real-time PCR of a segment of the X–Y homologous amelogenin ( AMG ) gene that allowed the simultaneous estimation of a Y-specific fragment (AMGY: 112 bp) and a X-specific fragment (AMGX: 106 bp) making possible not only haploid or diploid DNA quantitation but also sex determination. The AMG real-time PCR design has been used to quantify a set of 57 DNA samples from 4–5 years old forensic bone remains with improved sensitivity compared with the slot–blot hybridization method. The potential utility of this technology to improve the quality of some PCR-based forensic and ancient DNA studies (microsatellite typing and mtDNA sequencing) is discussed.

Polymorphisms at Major Histocompatibility Complex (MHC) genes have been associated with resistance/susceptibility to infectious diseases in domestic animals. The aim of this investigation was to evaluate whether polymorphisms of the DRA gene the Equine Lymphocyte Antigen is associated with susceptibility to Equine Arteritis Virus (EAV) infection in horses in Argentina. The equine DRA gene was screened for polymorphisms using Pyrosequencing® Technology which allowed the detection of three ELA-DRA exon 2 alleles. Neither allele frequencies nor genotypic differentiation exhibited any statistically significant (P-values = 0.788 and 0.745) differences between the EAV-infected and no-infected horses. Fisher’s exact test and OR calculations did not show any significant association. As a consequence, no association could be established between the serological condition and ELA-DRA. Fil: Kalemkerian, Paula Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Metz, German Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina Fil: Peral Garcia, Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Echeverria, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina Fil: Giovambattista, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina Fil: Diaz, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Genética Veterinaria "Ingeniero Fernando Noel Dulout"; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina

In this study, we have analysed the nucleotide sequence variation of the 12S rRNA mitochondrial gene (648–1601 bp) from five different populations (Spanish Caucasian, Autochthonous from the Basque Country, Chinese, New Guinea Highlander and Africans of Benin) by full sequencing of two overlapping PCR fragments using d-rhodamine cycle sequencing coupled with an ABI377 sequencer. Preliminary data indicate different patterns of sequence variation between Africans and non-Africans. Africans are much more polymorphic than non-African populations, which have only a very restricted subset of haplotypes. Furthermore, the greater part of Africans analysed showed two specific nucleotide substitutions (769G→A and 1018G→A) that were not observed in non-African individuals. In conclusion, the mtDNA 12SRNA gene in combination with other systems could be an interesting ethnic marker that could help to differentiate between African and non-African maternal lineages.

We here describe a multiplex–PCR method to generate six overlapping short amplicons (100–130 bp) in two separate PCR reactions of non-overlapping fragments for full sequencing of the whole hypervariable region I (HV1). The performance of this multiplex system has been evaluated not only on ancient bone remains (5000–4000 BP) but also on different forensic samples with highly degraded DNA (bone remains, hair shafts, …) that yielded negative PCR results with the mtDNA amplification strategies usually employed in forensic genetics. The multiplex–PCR methodology described in this study also illustrates a potential high throughput strategy to reconstruct the sequence of both coding and non-coding regions of the mtDNA genome from ancient DNA.

The sequence of the proximal upstream regulatory region (URR) of the bovine DRB genes was amplified using oligonucleotide primers designed from the consensus among DRB sequences from different species. The obtained DNA sequence was 234 bp long and composed of highly conserved sequence motifs, showing the same organization as the HLA-DRB, H2-IAb, H2-IEb and ELA-DRB genes. In this paper we report the sequence of the proximal upstream regulatory region (URR) of the bovine DRB genes. These are the first sequence data on the upstream regulatory regions of MHC class II genes in cattle. The reverse oligonucleotide primer (5'-GAGAAATACAGACACACCATGC-3') was designed from the consensus among DRB sequences from different species: BoLA-DRB (accession numbers U77067-68, U78548 and D45357), SLA-DRB (M55165-6), HLA-DRB (S72812 and L07838-40), H2-IEb and H2-IAb (X86151-6). As a forward primer we used the oligonucleotide proposed by Turco et al. (1990) (5'-TGTTTCAGAAAAGGACCTTC-3'), which was designed from the consensus among HLA-DRB sequences corresponding to the promoter region of the DRB genes. A polymerase chain reaction (PCR) was carried out in a total volume of 25 μl comprising 2.5 mM MgCl 2 , 20 mM Tris-HCl (pH = 8.4), 50 mM KCl, 100 μM of each dNTP, 0.5 μM of each primer, 1.0 unit Taq polymerase (Gibco BRL, Life-Technologies, Grand Island, NY) and 50-100 ng DNA template. The genomic DNA was extracted from whole blood from one animal of the Saavedreno Creole breed. The amplification profile consisted of 1 min at 94 °C, followed by 30 cycles of 45 s at 94 °C, 45 s at 55 °C and 45 s at 72 °C, with a final extension of 3 min at 72 °C. The amplification products were cloned into a dT-tailed pGEM-T easy vector (Promega, Madison, WI), and three clones of each PCR product were sequenced on an Applied Biosystems 377 automated sequencer (BioResource Center, Cornell University, Ithaca, NY), using a T7 universal primer. All the sequenced clones exhibited 100% sequence similarity to each other. The nucleotide sequence of the proximal upstream control region of the BoLA-DRB genes (GenBank accession number AY040327) was 234 bp long and composed of highly conserved sequence motifs that included, from the 5' to the 3' direction, W, X, Y, CCAAT and TATA-like boxes (Fig. 1), showing the same organization of the conserved regulatory elements as the HLA-DRB, H2-IAb, H2-IEb and ELA-DRB genes (e.g. Louis et al., 1993, 1994; Singal et al., 1993; Singal & Qiu, 1994). Furthermore, the BoLA-DRB URR nucleotide sequence had higher identity with HLA-DRB sequences than with HLA-DQB sequences (data not shown). This suggests that the sequence AY040327 corresponds to a BoLA-DRB promoter. However, we are still unable to assign this sequence to a specific BoLA-DRB gene, as at least three BoLA-DRB genes have been reported to exist (Andersson et al., 1986; Muggli-Cocket & Stone, 1988). BoLA-DRB3 is the most expressed DRB gene in cattle, while DRB1 is expressed at a low level and DRB2 is a pseudogene (Burke et al., 1991).

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 Veterinarias, Instituto de Genética Veterinaria

One of the major causes of embryo failure on in vitro-produced cattle embryos is their increased rate of chromosomal abnormalities. It was demonstrated that they can be increased by the pre- and post-environmental conditions of culture, such as the culture media and supplementation and culture atmosphere among others. Furthermore, it was suggested that the percentage of chromosomal abnormalities detected could be used as an indirect methodology to evaluate the embryo stress during culture. The use of oviduct cells in co-culture was employed as a technique that improves the embryo quality in different culture systems in cattle, probably by modulating the variations in the embryo environment or detoxifying the culture media. For that reason, we aimed to determine if the use of an oviducal co-culture system could reduce the percentage of chromosomal abnormalities in in vitro-produced cattle embryos. Oviducal cells were obtained from abattoir oviducts, and plated and cultured in TCM-199 media following standard procedures. Cumulus-oocyte complexes were also obtained from the abattoir and matured in a standard media (TCM199, with glutamine, pyruvate, FSH, LH, and antibiotics) for 20 h (38.5°C, 5% CO2). Matured oocytes were fertilized in IVF-TALP with 1 × 106 spermatozoa/mL (previously selected through a 45–90% Percoll™ gradient) for 18 h using the same conditions. Presumptive zygotes (n = 653) were divided into 2 groups and cultured in the mSof medium for 72 h, with (SOV) or without (SOF) the addition of oviducal cells. In total, 108 embryos were successfully karyotyped following our standard laboratory techniques. Chromosomal numerical abnormalities were evaluated by direct observation at 1250× magnification in a bright-field microscope using a simple Giemsa staining. The percentage of diploid embryos (54/58 in SOV and 47/53 in SOF) and abnormal embryos (4/58 in SOV and 6/53 in SOF) were nonsignificant (P > 0.05; Z-score test for 2 population proportions). Our results suggest that the use of oviducal cells as a co-culture supplementation in in vitro-produced cattle embryos do not improve the percentage of chromosomal abnormalities detected.

Genetic variation in the equine leucocyte antigen-DRB (ELA-DRB) second exon was investigated using polymerase chain reaction (PCR) amplification, restriction fragment length polymorphism (RFLP) of PCR products (PCR-RFLP) and deoxyribonucleic acid (DNA) sequencing. Eight distinct PCR-RFLP patterns could be identified in the studied Argentine Creole (AC) horses. The number of observed patterns per individual ranged from four to six, thus confirming the presence of multiple DRB copies in AC horses. Three PCR-RFLP alleles and three new sequences were identified. The estimated rates of synonymous and non-synonymous substitutions among ELA-DRB exon 2 sequences were higher within the antigen recognition site (ABS) than on the non-ABS. Phylogenetic analysis showed that the nucleotide sequences clustered in two main groups, while some sequences were not included in either group. Finally, the identification of the number of alleles per animal, the phylogenetic and segregation analyses allowed us to explain the number of ELA-DRB loci. However, it was not possible to identify specific alleles with specific loci.

Contrary to highly selected commercial breeds, indigenous domestic breeds are composed of semi-wild or feral populations subjected to reduced levels of artificial selection. As a consequence, many of these breeds have become locally adapted to a wide range of environments, showing high levels of phenotypic variability and increased fitness under natural conditions. Genetic analyses of three loci associated with milk production (alpha(S1)-casein, kappa-casein and prolactin) and the locus BoLA-DRB3 of the major histocompatibility complex indicated that the Argentinean Creole cattle (ACC), an indigenous breed from South America, maintains high levels of genetic diversity and population structure. In contrast to the commercial Holstein breed, the ACC showed considerable variation in heterozygosity (H(e)) and allelic diversity (A) across populations. As expected, bi-allelic markers showed extensive variation in He whereas the highly polymorphic BoLA-DRB3 showed substantial variation in A, with individual populations having 39-74% of the total number of alleles characterized for the breed. An analysis of molecular variance (AMOVA) of nine populations throughout the distribution range of the ACC revealed that 91.9-94.7% of the total observed variance was explained by differences within populations whereas 5.3-8.1% was the result of differences among populations. In addition, the ACC breed consistently showed higher levels of genetic differentiation among populations than Holstein. Results from this study emphasize the importance of population genetic structure within domestic breeds as an essential component of genetic diversity and suggest that indigenous breeds may be considered important reservoirs of genetic diversity for commercial domestic species.