A pregnancy diagnosis is an important standard for control of livestock’s reproduction in paricular dairy cattle. High reproductive performance in dairy animals is a essential condition to realize of high life-time production. Pregnancy diagnosis is crucial to shortening the calving interval by enabling the farmer to identify open animals so as to treat or re-breed them at the earliest opportunity. MicroRNAs are short RNA molecules which are critically involved in regulating gene expression during both health and disease. This study is sought to establish the feasible of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from 12 non-pregnant cows (“open” cows: samples were collected before insemination (non-pregnant state) and after pregnancy check at the indicated time points) on weeks 0, 4, 8, 12 and 16. Using small RNA sequencing we identified a total of 115 miRNAs that were differentially expressed weeks 16 relative to non-pregnancy (“open” cows). Weeks 8, 12 and 16 of pregnancy commonly showed a distinct increase in circulating levels of miR-221 and miR-320a. Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with pregnancy in cattle. Their application in the field of reproductive biology has opened up opportunities for research communities to look for pregnancy biomarker molecules in dairy cattle.

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
CONCLUSION
CONFLICTS OF INTEREST
ACKNOWLEDGEMENTS
AUTHOR CONTRIBUTIONS
AUTHOR’S POSITION AND ORCID NO
REFERENCES

• Hyun-Joo Lim(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea) Corresponding author
• Hyun Jong Kim(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea)
• Ji Hwan Lee(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea)
• Dong Hyun Lim(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea)
• Jun Kyu Son(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea)
• Eun-Tae Kim(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea)
• Gulwon Jang(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea)
• Dong-Hyeon Kim(Dairy Science Division, Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 31000, Korea)