Profiling and identification of pregnancy-associated circulating microRNAs in dairy cattle.

Holstein is one among the dairy cattle which provide higher milk yields than most other cattle breeds. Lack of high-accuracy, reliable methods for early detection of cattle pregnancy reduces overall productivity and constitutes a high economic burden to the dairy industry. The circulating microRNAs (miRNAs) in exosomes could provide information and serve as potential biomarkers for livestock health and disease. However, the complexity of miRNA in response to cattle early pregnancy remains unknown. Hence, we collected blood samples of three healthy dairy cows of normal and 30 days of pregnancy, in order to further characterize the miRNA transcriptome profile. A high-throughput RNA-Seq approach detected 794 known and 2154 novel circulating miRNAs in six libraries. A total of 29 miRNAs in the 30 days of pregnancy group showed significant differences compared to the normal group. Further, bta-miR-450b, bta-miR-146b, bta-miR-26b and bta-miR-27b were up-regulated which shown to be involved in preeclampsia, immune response and mammary gland development. GO enrichment analysis showed these target genes were involved in the metabolic process, signal transducer activity, and membrane etc., while KEGG analysis showed that these genes were enriched in membrane trafficking, chromosome and associated proteins, exosome and G protein-coupled receptors pathways. These results provide an experimental basis to reveal the potential role of miRNAs as biomarkers in early diagnosis of pregnancy and other molecular functions.

Genome-wide association study and prediction of genomic breeding values for fatty-acid composition in Korean Hanwoo cattle using a high-density single-nucleotide polymorphism array.

Genomic selection using high-density single-nucleotide polymorphism (SNP) markers is used in dairy and beef cattle breeds to accurately estimate genomic breeding values and accelerate genetic improvement by enabling selection of animals with high genetic merit. This genome-wide association study (GWAS) aimed to identify genetic variants associated with beef fatty-acid composition (FAC) traits and to evaluate the accuracy of genomic predictions (GPs) for those traits using genomic best linear unbiased prediction (GBLUP), pedigree BLUP (PBLUP), and BayesR models. Samples of the longissimus dorsi muscle of 965 thirty-month-old Hanwoo steers (progeny of 73 proven bulls) were used to investigate 14 FAC traits. Animals were genotyped or imputed using two bovine SNP platforms (50K and 777K), and after quality control, 38,715 (50K) and 633,448 (777K) SNPs were subjected to GWAS and GP study using a cross-validation scheme. SNP-based heritability estimates were moderate to high (0.25 to 0.47) for all studied traits, with some exceptions for polyunsaturated fatty acids. Association analysis revealed that 19 SNPs in BTA19 (98.7 kb) were significantly associated (P < 7.89 × 10-8) with C14:0 and C18:1n-9; these SNPs were in the fatty-acid synthase (FASN) and coiled-coil domain-containing 57 (CCDC57) genes. BayesR analysis revealed that 0.41 to 0.78% of the total SNPs (n = 2,571 to 4,904) explained almost all of the genetic variance; the majority of the SNPs (>99%) had negligible effects, suggesting that the FAC traits were polygenic. Genome partitioning analysis indicated mostly nonlinear and weak correlations between the variance explained by each chromosome and its length, which also reflected the considerable contributions of relatively few genes. The prediction accuracy of breeding values for FAC traits varied from low to high (0.25 to 0.57); the estimates using the GBLUP and BayesR methods were superior to those obtained by the PBLUP method. The BayesR method performed similarly to GBLUP for most of the studied traits but substantially better for those traits that were controlled by SNPs with large effects; this was supported by the GWAS results. In addition, the predictive abilities of the 50K and 777K SNP arrays were almost similar; thus, both are suitable for GP in Hanwoo cattle. In conclusion, this study provides important insight into the genetic architecture and predictive ability of FAC traits in Hanwoo cattle. Our findings could be used in selection and breeding programs to promote production of meat with enhanced nutritional value.

Effects of Medicinal Herb Extracts on In vitro Ruminal Methanogenesis, Microbe Diversity and Fermentation System.

This study was aimed to evaluate the in vitro effects of medicinal herb extracts (MHEs) on ruminal fermentation characteristics and the inhibition of protozoa to reduce methane production in the rumen. A fistulated Hanwoo was used as a donor of rumen fluid. The MHEs (T1, Veratrum patulum; T2, Iris ensata var. spontanea; T3, Arisaema ringens; T4, Carduus crispus; T5, Pueraria thunbergiana) were added to the in vitro fermentation bottles containing the rumen fluid and medium. Total volatile fatty acid (tVFA), total gas production, gas profiles, and the ruminal microbe communities were measured. The tVFA concentration was increased or decreased as compared to the control, and there was a significant (p<0.05) difference after 24 h incubation. pH and ruminal disappearance of dry matter did not show significant difference. As the in vitro ruminal fermentation progressed, total gas production in added MHEs was increased, while the methane production was decreased compared to the control. In particular, Arisaema ringens extract led to decrease methane production by more than 43%. In addition, the result of real-time polymerase chain reaction indicted that the protozoa population in all added MHEs decreased more than that of the control. In conclusion, the results of this study indicated that MHEs could have properties that decrease ruminal methanogenesis by inhibiting protozoa species and might be promising feed additives for ruminants.

The In vitro Effects of Nano-encapsulated Conjugated Linoleic Acid on Stability of Conjugated Linoleic Acid and Fermentation Profiles in the Rumen.

This study was aimed to evaluate the stability of conjugated linoleic acids (CLAs) by nano-encapsulation against in vitro ruminal biohydrogenation by microbial enzymatic conversion. CLAs (free fatty acid form of CLA [CLA-FFA], nano-encapsulated CLA-FFA, triglyceride form of CLA [CLA-TG], and nano-encapsulated CLA-TG) were used in the in vitro fermentation experiments. When Butyrivibrio fibrisolvens (B. fibrisolvens) was incubated with CLA-FFAs, the concentrations of cis-9, trans-11 CLA and vaccenic acid (VA) slightly was decreased and increased by nano-encapsulation, respectively. When B. fibrisolvens was incubated with CLA-TG, the concentrations of cis-9, trans-11 CLA and VA decreased, but these were increased when B. fibrisolvens was incubated with nano-encapsulated CLA-TG. The nano-encapsulation was more effective against the in vitro biohydrogenation activity of B.fibrisolvens incubated with CLA-FFA than with CLA-TG. In the in vitro ruminal incubation test, the total gas production and concentration of total volatile fatty acids incubated with nano-encapsulated CLA-FFA and CLA-TG were increased significantly after 24 h incubation (p<0.05). Nano-encapsulated CLA-FFA might, thus, improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, nano-encapsulated CLA-FFA increased the population of Fibrobacter succinogenes and decreased the population of B. fibrisolvens population. These results indicate that nano-encapsulation could be applied to enhance CLA levels in ruminants by increasing the stability of CLA without causing adverse effects on ruminal fermentation.

Deciphering the genetic blueprint behind Holstein milk proteins and production.

Holstein is known to provide higher milk yields than most other cattle breeds, and the dominant position of Holstein today is the result of various selection pressures. Holstein cattle have undergone intensive selection for milk production in recent decades, which has left genome-wide footprints of domestication. To further characterize the bovine genome, we performed whole-genome resequencing analysis of 10 Holstein and 11 Hanwoo cattle to identify regions containing genes as outliers in Holstein, including CSN1S1, CSN2, CSN3, and KIT whose products are likely involved in the yield and proteins of milk and their distinctive black-and-white markings. In addition, genes indicative of positive selection were associated with cardiovascular disease, which is related to simultaneous propagation of genetic defects, also known as inbreeding depression in Holstein.