Differential gene expression in tilapia Oreochromis niloticus (L., 1758) under feeding stress

Feeding stress or suboptimal feeding in fish can have a direct impact on their energy metabolism and feed use efficiency, and thus on productivity. That impact can be quantified objectively by comparing the expression of genes between fish fed ad libitum and fish fed suboptimally. Moreover, some of the genes expressed in animals fed suboptimally can also be used as candidate genes to identify feeding stress in general. To identify some of those genes, a group of 60 tilapia Oreochromis niloticus (L., 1758) was divided into two batches (fed ad libitum or suboptimally for 22 days) with two replicates. For this purpose, we carried out differential display analysis in brain tissue samples using 36 combinations of 3 polyT anchoring primers and 12 random primers. An average of 25 bands was produced per primer pair, with 31 bands expressed differentially which were excised from the gels, sequenced, and annotated with information available on public databases. Some sequences matched genes with different binding activities, regulation of transcription, and those playing a role in synapses. One of the most interesting findings was the ALOX5 gene, which plays a role in the production of lukotrienes, important mediators in inflammatory processes. The identification of genes corresponding to those bands will provide more information about mechanisms that play a role in stressful situations produced by suboptimal feeding, as well as other causes.El estrés alimentario o subalimentación de los peces afecta directamente al metabolismo energético, a la eficacia de utilización del alimento y, por tanto, a su productividad. Este efecto se puede cuantificar de forma objetiva mediante comparaciones de expresión génica con respecto a aquellos peces que han sido alimentados a saciedad. Además, algunos de los genes expresados por los peces subalimentados pueden usarse como genes candidatos para identificar estrés alimentario en general. Para identificar estos genes se dividió un grupo de 60 tilapias Oreochromis niloticus (L., 1758) en dos tratamientos (alimentado ad líbitum o con alimentación subóptima, durante 22 días) con dos réplicas. Se utilizó la técnica de differential display en muestras de cerebro con 36 combinaciones resultado de tres cebadores polyT de anclaje y 12 cebadores aleatorios. Se expresaron una media de 25 bandas por pareja de cebadores y un total de 31 bandas expresadas diferencialmente, se extrajeron, secuenciaron y anotaron con la ayuda de la información disponible en bases de datos públicas. Algunas secuencias coinciden con genes cuya función es de unión a diferentes proteínas y de regulación de la transcripción, y juegan un papel importante en la sinapsis. Es destacable el gen ALOX5, que interviene en la producción de leucotrienos, importantes mediadores de procesos inflamatorios. La identificación de los genes que corresponden a estas bandas aportará conocimiento sobre los mecanismos que intervienen en situaciones de estrés, tanto producidas por una alimentación subóptima como por otras causas.Instituto Español de Oceanografí

DGAT1 p.K232A polymorphism in dairy and dual purpose Italian cattle breeds

The aim of this study was to evaluate the allele frequency distribution at the DGAT1 p.K232A polymorphic site in seven Italian dairy and dual purpose cattle breeds. On the whole, 651 animals belonging Italian Holstein (116), Italian Brown (105), Italian Simmental (95), Valdostana Red Pied (95), Rendena (62), Reggiana (128) and Modenese (50) were genotyped by PCR-RFLP. Sequencing was carried out to confirm results of the genotyping protocol. The DGAT1 p.232K allele was identified in Italian Holstein (25.4%), Reggiana (17.2%), and with very low frequency in Italian Simmental, Valdostana Red Pied and Rendena (1%). In Italian Brown and Modenese, this allele was not detected. These results indicated that this polymorphic site can be considered for association studies only in Italian Holstein and Reggiana breeds. Deviation from Hardy-Weinberg equilibrium was observed in the Reggiana breed (P<0.01) in which there was an excess of heterozygous sires and absence of animals with the p.232KK genotype. This result should be further evaluated because the analysed sires represented almost all bulls available for artificial insemination in this breed. Comparison of allele frequencies at the DGAT1 locus with several other Holstein populations showed a wide range of variability, probably due to different selection strategies adopted