Feeding olive cake silage up to 20% of DM intake in sheep improves lipid quality and health-related indices of milk and ovine halloumi cheese.

This study aimed to evaluate the use of a by-product, olive cake silage (OCS), as a forage replacement in sheep diets for the improvement of fatty acid (FA) content of milk and thus, the lipids of the ovine halloumi cheese produced. Sixty second-parity purebred Chios ewes in mid-lactation were assigned to three diet treatments (2 lots of 10 animals per treatment) receiving 0%, 10%, and 20% of OCS on dry matter basis for 3 weeks (treatments S0, S10, and S20, respectively). Halloumi cheese was manufactured from fresh raw milk of ewes fed the three different diets. Inclusion of OCS in the diets increased linearly the concentration in milk of unsaturated FA up to 20%, monounsaturated FA up to 23%, polyunsaturated FA up to 11%, rumenic acid (CLA cis-9, trans-11) up to 61%, and consequently reduced the atherogenicity and thrombogenicity milk indices by 31% and 27%, for the S10 and S20 treatments, respectively, compared with the control treatment. Moreover, these differences were carried over to the lipid profile of ovine halloumi cheese showing, on average, more than 25% increase of unsaturated, polyunsaturated, and monounsaturated FA, with particularly enhanced oleic and rumenic acid content. These changes resulted in reduced atherogenicity by 29% and 45% and thrombogenicity by 23% and 24% of ovine halloumi cheese made from milk of S10 and S20 diets, respectively. Milk yield, milk fat, or protein content was not affected by S10 or S20 feeding treatments compared to control. Overall, the applied ensiling method of olive cake produces a by-product that can be included as a forage replacement up to 20% of DM intake in Chios sheep without adversely affecting the lactating performance. Furthermore, the present study showed that such substitution improves the lipid quality of milk and related halloumi cheese enriching these ovine dairy products with beneficial to human health fatty acids.

The use of ensiled olive cake in the diets of Friesian cows increases beneficial fatty acids in milk and Halloumi cheese and alters the expression of SREBF1 in adipose tissue.

This study aimed to evaluate the effect of dietary inclusion of ensiled olive cake, a by-product of olive oil production, on milk yield and composition and on fatty acid (FA) profile of milk and Halloumi cheese from cows. Furthermore, the effect of olive cake on the expression of selected genes involved in mammary and adipose lipid metabolism was assessed in a subset of animals. A total of 24 dairy cows in mid lactation were allocated into 2 isonitrogenous and isoenergetic feeding treatments, named the control (CON) diet and the olive cake (OC) diet, in which part of the forages (alfalfa, barley hay, and barley straw) were replaced with ensiled OC as 10% of dry matter according to a 2 × 2 crossover design with two 28-d experimental periods. At the end of the second experimental period, mammary and perirenal adipose tissue samples were collected from 3 animals per group for gene expression analysis by quantitative reverse-transcription PCR. The expression of 11 genes, involved in FA synthesis (ACACA, FASN, G6PDH), FA uptake or translocation (VLDLR, LPL, SLC2A1, CD36, FABP3), FA saturation (SCD1), and transcriptional regulation (SREBF1, PPARG), was evaluated. No significant differences were observed between groups concerning milk yield, fat percentage, protein percentage, and protein yield (kg/d), whereas milk fat yield (kg/d) increased in the OC group. Dietary supplementation with ensiled OC modified the FA profile of milk and Halloumi cheese produced. There was a significant decrease in the concentration of de novo synthesized FA, saturated FA, and the atherogenic index, whereas long-chain and monounsaturated FA concentration was increased in both milk and cheese. Among individual saturated FA, only stearic acid was elevated, whereas among individual monounsaturated FA, increments of oleic acid (C18:1 cis-9) and the sum of C18:1 trans-10 and trans-11 acids were demonstrated in milk and Halloumi cheese produced. Although no diet effect was reported on total polyunsaturated FA, the concentration of CLA cis-9,trans-11 was increased in both milk and Halloumi cheese fat of the OC group. The expression of the genes tested was unaffected apart from an observed upregulation of SREBF1 mRNA expression in perirenal fat from cows fed the OC diet. Milk FA differences observed were not associated with alterations in mammary expression of genes involved in FA synthesis, uptake, translocation, and regulation of lipogenesis. Overall, the inclusion of ensiled OC in cow diets for a 4-wk period improved, beneficially for human health, the lipid profile of bovine milk and Halloumi cheese produced without adversely affecting milk yield and composition or the expression of genes involved in lipid metabolism of mammary and adipose tissues in cows.

Variants in the 3' untranslated region of the ovine acetyl-coenzyme A acyltransferase 2 gene are associated with dairy traits and exhibit differential allelic expression.

The acetyl-CoA acyltransferase 2 (ACAA2) gene encodes an enzyme of the thiolase family that is involved in mitochondrial fatty acid elongation and degradation by catalyzing the last step of the respective ß-oxidation pathway. The increased energy needs for gluconeogenesis and triglyceride synthesis during lactation are met primarily by increased fatty acid oxidation. Therefore, the ACAA2 enzyme plays an important role in the supply of energy and carbon substrates for lactation and may thus affect milk production traits. This study investigated the association of the ACAA2 gene with important sheep traits and the putative functional involvement of this gene in dairy traits. A single nucleotide substitution, a T to C transition located in the 3' untranslated region of the ACAA2 gene, was used in mixed model association analysis with milk yield, milk protein yield and percentage, milk fat yield and percentage, and litter size at birth. The single nucleotide polymorphism was significantly associated with total lactation production and milk protein percentage, with respective additive effects of 6.81 ± 2.95 kg and -0.05 ± 0.02%. Additionally, a significant dominance effect of 0.46 ± 0.21 kg was detected for milk fat yield. Homozygous TT and heterozygous CT animals exhibited higher milk yield compared with homozygous CC animals, whereas the latter exhibited increased milk protein percentage. Expression analysis from age-, lactation-, and parity-matched female sheep showed that mRNA expression of the ACAA2 gene from TT animals was 2.8- and 11.8-fold higher in liver and mammary gland, respectively. In addition, by developing an allelic expression imbalance assay, it was estimated that the T allele was expressed at an average of 18% more compared with the C allele in the udder of randomly selected ewes. We demonstrated for the first time that the variants in the 3' untranslated region of the ovine ACAA2 gene are differentially expressed in homozygous ewes of each allele and exhibit allelic expression imbalance within heterozygotes in a tissue-specific manner, supporting the existence of cis-regulatory DNA variation in the ovine ACAA2 gene. This is the first study reporting differential allelic imbalance expression of a candidate gene associated with milk production traits in dairy sheep.

Short communication: Identification of variation in the ovine prolactin gene of Chios sheep with a cost-effective sequence-based typing assay.

The present study identified single nucleotide polymorphisms (SNP) in the coding and untranslated regions of the ovine prolactin gene of Chios sheep. By developing a cost-effective direct sequence-based typing assay, around 600 bp of reliable sequencing data and clear identification of heterozygous positions was achieved. Five SNP were found, located in exons 2 (KC764410:g.567G>A, g.625C>T, g.683C>A) and 3 (KC764410:g.2015C>A, g.2101G>A), whereas the remaining exons were monomorphic. The identified SNP were synonymous, with the exception of the g.567G>A SNP, which results in an Arg to His amino acid change. As the sequencing cost of the sequence-based typing assay was 20 orders of magnitude lower compared with a standard Sanger method, the assay was also used as a genotyping tool. The identified polymorphism was genotyped for 247 ewes and was subsequently used in mixed model association analyses of milk yield, milk fat content, and litter size at birth. The association analysis revealed a significant dominance effect of 0.17 ± 0.07 of the g.2015C>A SNP on milk fat percentage, whereas a dominance effect of -21.33 ± 10.51 of the same SNP on total lactation milk yield was also estimated. The g.2015C>A SNP explained 2.47 and 3.68% of the total phenotypic variance of milk yield and milk fat percentage, respectively, whereas the corresponding values for the animal variance were 7.14 and 11.75%. A suggestive association of the nonsynonymous g.567G>A SNP with litter size at birth was also detected.