Early Pregnancy In Cattle Research Articles

RNA Sequencing Reveals the Involvement of Serum Exosomal miRNAs in Early Pregnancy in Cattle.

Low fertility is the main cause of the low productivity in beef cattle and is mainly associated with a lack of conception after fertilization. The establishment of early pregnancy in cattle is a complex physiological process, and embryo implantation is crucial for the successful establishment of pregnancy. Exosomal miRNAs play an important role in regulating mammalian embryo implantation and development. This study used synchronous estrus technology to extract exosomes from bovine serum at 0, 14, and 21 days of early pregnancy and analyzed the expression profile of exosomal miRNAs through RNA-seq technology. We identified 472 miRNA precursor sequences and 367 mature miRNA sequences in the three sample groups, with the majority of the miRNAs having high abundance. Differentially expressed miRNAs (DEmiRNAs) were screened, and 20 DEmiRNAs were obtained. The differential expression analysis results show that compared to day 0, there were 15 DEmiRNAs in the serum on day 14 and 5 on day 21 of pregnancy. Compared to the 14th day of pregnancy, there were eight DEmiRNAs in the serum on the 21st day of pregnancy. Bioinformatics analysis shows that the target genes of DEmiRNAs regulated the signaling pathways closely related to early pregnancy, including the VEGF, NF-κB, and MAPK signaling pathways. In addition, the newly discovered miRNAs were bta-miR-3604, bta-miR-2889, bta-miR-3432a, and bta-miR-409b. These results provide a theoretical reference for screening the molecular markers for early pregnancy establishment and maternal recognition of pregnancy (MRP) in cattle and new ideas for shortening the calving interval in cows.

Potential of Circulating miRNA Biomarkers and Exosomes for Early Pregnancy Diagnoses in Cattle.

Circulating microRNAs (miRNAs) were investigated as biomarkers for the diagnosis of early pregnancy in cattle. The levels of prospective miRNA biomarkers and the features of extracellular vesicles (EVs) in the blood were evaluated. In Study 1, plasma samples from cows 21 days after artificial insemination (AI) were examined using RT-qPCR to determine the levels of seven circulating miRNAs. Only the levels of miR-126-3p were significantly lower in the pregnant group than in the non-pregnant group. In Study 2, among individuals not pregnant at the first AI, the miRNA levels were compared between the individuals pregnant at the second AI and those who remained non-pregnant. The miR-25 levels were significantly higher in the pregnant group at the second AI than in the pregnant group at the first AI; miR-19b, miR-27b, and miR-29a levels were also high. In the non-pregnant group, changes were absent in the miRNA levels in the same individual between the first and second AIs. In Study 3, Western blotting and RT-qPCR showed the presence of miRNAs in EVs and their levels were lower than in plasma. Thus, circulating miR-126-3p may serve as a biomarker for the diagnosis of early pregnancy in cattle. In addition, the expression of some miRNAs tended to be higher during pregnancy than during non-pregnancy in the same individual, suggesting their potential as an index to determine pregnancy and non-pregnancy rates using a comparative method.

MX2 gene mRNA expression as potential biomarker for early pregnancy diagnosis in cattle

Early pregnancy diagnosis is vital for economic sustainability of dairy farms and maintaining the reproductive efficiency of the herd. There are many techniques including progesterone assay, pregnancy specific proteins and interferon stimulated genes have been explored for early pregnancy diagnosis but, they are associated with varying level of efficacy. In the present experiment, interferon stimulated gene (Myxovirus resistance gene 2/MX2) expression pattern was used as a potential biomarker for early pregnancy in cattle. The association of MX2 gene expression in relation to progesterone assay was studied to explore its potential use as biomarker of early pregnancy. The plasma progesterone concentration in conceived animals on day 7 (2.26±0.19 ng/ml), 17 (5.42±0.35 ng/ml) and 21(6.38±0.39 ng/ml) was recorded to be significantly higher as compared to respective values in non-conceived animals, i.e. 1.55±0.09 ng/ml, 4.14±0.14 ng/ml and 0.81±0.06 ng/ml. The sudden decrement in plasma progesterone concentration after day 17th discriminates conceived and non-conceived animals. MX2 expression levels were observed to spike in blood due to release of interferon tau (τ) after implantation of embryo. The relative mRNA expression of MX2 gene showed a 9.5 to 28.64-fold higher expression on 17 days post insemination in pregnant animals as compared to non-pregnant animals. Thus, MX2 gene can be used as a reliable biomarker for the early detection of pregnancy.

Effects of apoptosis by 20α-hydroxysteroid dehydrogenase activity on corpus luteum formation during early pregnancy in cattle

20α-hydroxysteroid dehydrogenase (20α-HSD) is a member of the aldo-keto reductase family. These enzymes play a pivotal role in regulating steroid hormones, such as androgens, oestrogen, and progesterone, and are, therefore, considered vital targets for determining whether a pregnancy is maintained. In this study, we investigated the association between 20α-HSD and apoptosis-related genes in luteal tissues (at 30, 60, and 90 days of gestation) and cells from early gestation in cattle. The corpus luteum altered the number of large lutein cells from 30–90 days. The change in the junction of the connective tissue appeared to affect the density of the corpus luteum. In addition, 20α-HSD was detected in the corpus luteum and cultured cells during early pregnancy, in contrast to the results of previous studies. The overall expression pattern of the 20α-HSD and Casp-3 proteins was lower on day 50 of gestation than on days 30 and 90. However, the 20α-HSD expression gradually increased from 30 to 90 days of gestation. When the 20α-HSD protein was increased to 0.5 μg/ml, 1 μg/ml, and 1.5 μg/ml in the luteal cells collected on day 30 of pregnancy, apoptosis was analysed after 48 hours. 20α-HSD generated in the cells was confirmed, and the concentration gradually decreased as the concentration increased. However, the expression of Casp-3 showed an overall similar expression pattern. Notably, the 20α-HSD and Casp-3 proteins were lowest at 1.5 μg/ml supplemented with 20α-HSD, with higher levels in the cytosol than in the cytoplasm. These results suggest that 20α-HSD plays a role in maintaining normal pregnancy, particularly by regulating the progesterone concentrations during luteal cell development.

Short communication: profiling the expression of Let-7d-5p microRNA in circulating blood of pregnant and nonpregnant cows.

The objective of this work was to determine if specific circulating microRNA (miRNA) differed due to pregnancy status in heifers. Blood samples were collected from heifers 21 d after receiving an in vitro-produced embryo. Pregnancy status was diagnosed 21 d after embryo transfer, equivalent to day 28 of gestation, with rectal ultrasonography. Blood samples from 10 pregnant and 10 nonpregnant heifers were then evaluated for miRNA expression. There were five different miRNAs quantified using delta-delta Ct and qPCR methodology. These miRNAs had previously been associated with early pregnancy in cattle. The miRNA Let-7d-5p was decreased in nonpregnant as compared to pregnant females (P < 0.05). There were no changes in 16-5p, 16-1-3p, 16-2-3p, and 26a-5p associated with pregnancy (P > 0.05). Results demonstrate an opportunity to identify and study the differential expression of miRNAs from the blood of pregnant cows. The Let-7d-5p miRNA is a potential early pregnancy marker and is critical to better understand the early relationships of the cellular and molecular interactions of the cow and embryo.

Ruminant conceptus-maternal interactions: interferon-tau and beyond.

Embryonic or fetal loss in cattle is associated with problems that occur during oocyte maturation, early embryonic development, conceptus elongation, maternal recognition of pregnancy (MRP), and/or placental attachment and implantation. Many of these problems manifest as inadequate or asynchronous communication between the developing conceptus and endometrium, resulting in pregnancy failure. This review will provide an overview of how various conceptus-endometrial paracrine signaling systems control the fate of early pregnancy in cattle and other ruminants. We begin by summarizing the actions of interferon-tau, the classic MRP signal in ruminates, and then explore how other secretory factors derived from either the conceptus or endometrium influence establishment and maintenance of pregnancy. Insight into how the endometrium responds to male vs. female conceptuses or conceptuses produced by in vitro methods will also be described. Specific focus will be placed on describing how "omic" technologies and other cutting-edge techniques have assisted with identifying novel conceptus and/or endometrial factors and their functions. Recent findings indicate that the endometrial transcriptome and histotroph are altered by conceptus sex, quality, and origin, suggesting that the endometrium is a sensor of conceptus biochemistry. Although the endometrium has a certain level of flexibility in terms of conceptus-maternal interactions, this interplay is not sufficient to retain some pregnancies. However, new information inspires us to learn more and will help develop technologies that mitigate early embryonic loss and reproductive failure in ruminants and other animals.

A Comparative Study on Certain Blood Metabolites and Hormonal Indices for Detection of Pregnancy of Crossbred Cows of High Altitude: Mizoram

Background: Early detection of pregnancy can perk up the reproductive competence of dairy animals and decreasing the interval between two successive AI. Several methods of pregnancy diagnosis both direct and indirect are being practiced; but to date, none of these technologies qualifies as the ideal pregnancy detection method due to its limitation. The present study was designed to learn about the changes of hormonal and blood metabolites during early bovine pregnancy. Methods: A total of 30 numbers of crossbred cows at 2nd to 5th lactation was selected for the study. Clinico-gynaecological examination was carried out in all the cows and those have devoid of ill health, having a regular estrous cycle and estrus period were inseminated at mid to late estrus. Ten milliliter of blood samples were collected aseptically from all the cows (6 samples from each cow and a total of 180 samples) on day 1, 10, 20, 30, 60 and 90 of post-A.I. From the obtained serum, levels of glucose, cholesterol, BUN, total protein and hormonal profile estrone sulphate, progesterone were analysed. All the cows were observed for the absence of external signs of estrus after AI and pregnancy was confirmed by using ultrasonography and per rectal examination on day 30 to 40 of post insemination. Result: It was observed that level of glucose, BUN, cholesterol and progesterone was significantly increased during early pregnancy (P less than 0.01). While, the serum level of estrone sulphate significantly decreased while advancing the pregnancy. There was positive correlation between the level of progesterone (P less than 0.01) and negative correlation of estrone sulphate (P less than 0.05) with serum glucose, BUN, cholesterol at different days of post A.I. The blood biochemical constituents i.e. glucose, BUN, cholesterol and hormone estrone sulphate, progesterone possibly will used as an ideal marker of early pregnancy in cattle. Thus blood biochemical constituents like glucose, BUN, cholesterol and hormone estrone sulphate, progesterone possibly can be used as a tool for diagnosis of non pregnant dairy cattles with very high accuracy at around 30 days of post insemination.

Selection of early pregnancy specific proteins and development a rapid immunochromatographic test strip in cows

The most of embryo losses occur before the day 16 after artificial insemination, but there is no low cost and easy operation that can detect pregnancy with high accuracy within three weeks post-insemination in cattle. In this study, blood samples were collected at day 18 of the estrous cycle, and days 18, 25 and 35 of pregnancy, and relative levels of interferon stimulated genes (ISGs), Toll-like receptor (TLRs), complement components, early pregnancy factor (EPF) and pregnancy-associated plasma protein A (PAPPA) proteins were analyzed through Western blot. In addition, a colloidal gold immunochromatographic test strip was developed using the selected antibody, and the test was used for early pregnancy diagnosis. The results showed that there were changes in relative levels of plasma ISGs, TLRs, complement components, EPF and PAPPA proteins during early pregnancy in cattle, and complement component 1q (C1q) could be used as an ideal marker to develop a colloidal gold immunochromatographic test strip for early pregnancy diagnosis. In addition, the accuracy of pregnancy diagnosis by this test strip was 91.67% (11/12) for pregnant cows and 80% (8/10) nonpregnant cows at day 18 after insemination. In conclusion, the changes in plasma ISGs, TLRs, complement components, EPF and PAPPA proteins may be related to the maternal systemic immune modulation during early pregnancy, and a colloidal gold immunochromatographic test strip was developed for early pregnancy diagnosis using C1q as the ideal marker in cows. However, this colloidal gold immunochromatographic test strip needs further studies to improve the accuracy.

Role of reproductive fluids and extracellular vesicles in embryo&#x2013;maternal interaction during early pregnancy in cattle.

The coordinated interaction between the developing embryo and the maternal reproductive tract is essential for the establishment and maintenance of pregnancy in mammals. An early cross-talk is established between the oviduct/uterus and the gametes and embryo. This dialogue will shape the microenvironment in which gamete transport, fertilisation, and early embryonic development occur. Due to the small size of the gametes and the early embryo relative to the volume of the oviductal and uterine lumina, collection of tissue and fluid adjacent to these cells is challenging in cattle. Thus, the combination of in vivo and in vitro models seems to be the most appropriate approach to better understand this fine dialogue. In this respect, the aim of this review is to summarise the recent findings in relation to gamete/embryo-maternal interaction during the pre-elongation period.

EVALUATION OF SULFURIC ACID, BARIUM CHLORIDE, AND SEED GERMINATION ASSAY METHODS AS EARLY PREGNANCY DETECTION INSTRUMENTS IN CATTLE

This study aims to validate and re-evaluate the use of H2SO4, BaCl2, and seed germination assay (SGA) methods to detect early pregnancy in cattle. Validation was carried out in order to find out the actual test performance value, which include accuracy, sensitivity, and specificity.Validation test was done by reacting the urine of pregnant and non-pregnant cattle populations with H2SO4 solution diluted at a ratio of 1 : 4, with 1% BaCl2 solution or through germination of mung bean seeds in Petri dishes. The results of this study indicated that the H2SO4 and BaCl2 methods have poor accuracy (21.88% and 28.13%) with the sensitivity of 81.82% and 100% and specificity of 9.43% and 13.21%, respectively. Meanwhile, the SGA method had a fairly good accuracy (74.55%) with sensitivity and specificity of 45.46% and 81.82%, respectively. Based on these results, the H2SO4, BaCl2, and SGA methods are not recommended for detection of early pregnancy in the field.

Bovine scavenger receptor class A (SR-A) exhibit specific patterns of regulation in the endometrium during the oestrous cycle and early pregnancy.

In mammals, tight regulation of maternal endometrial function is critical for pregnancy success. In bovine species, endometrial expression of members of the scavenger receptor class A (SR-A) has been listed in high-throughput analyses, but very little is known about the involvement of these immune factors during implantation in mammals. To provide first insights into the contribution of SR-A to endometrial physiology, we analysed the expression and regulation of all members of SR-A (SR-A1, SR-A3-SR-A6) during the oestrous cycle and early pregnancy in cattle. Levels of SR-A1 were increased on Day 20 of pregnancy, whereas SR-A3 levels were increased on Day 13 of the oestrous cycle and of the pregnancy. Although SR-A4 levels were reduced on Day 20 of the oestrous cycle, they remained high in pregnant animals. SR-A5 levels increased by Day 13 of the oestrous cycle and decreased on Day 20, but remained high in pregnant animals. Interferon-τ does not affect SR-A gene expression, whereas progesterone regulates the expression of the SR-A3 and SR-A5 transcripts. Endometrial SR-A3 appeared significantly higher in cows carrying invitro-produced embryos than in AI cows. Our data suggest that members of the SR-A family are involved in endometrial remodelling and regulation of endometrial gland physiology, both processes being critical for implantation in mammals.

Regulatory Role of Local Immune Response During the Early Pregnancy in Cattle

Regulatory Role of Local Immune Response During the Early Pregnancy in Cattle

Exploring maternal serum microRNAs during early pregnancy in cattle

Confirmation of the pregnancy establishment at the very earliest day post-insemination increases the reproduction efficiency of high yielding dairy cows and farm profitability by allowing rebreeding of the non-pregnant cows. Inaccuracies in the currently available pregnancy detection tools to detect pregnancy establishment within the first 3 weeks post insemination extends the inter-calving interval and have contributed to the decline in profitability. Thus, development of non-invasive early pregnancy detection biomarkers could be proposed as alternative tools. MicroRNAs (miRNAs), a subclass of small non-coding RNAs are abundantly expressed in virtually all bio fluids circulation and have been associated with various pregnancy-related pathophysiological conditions. The study aimed to determine the expression of circulatory miRNAs in serum samples of pregnant and non-pregnant cows at day 19 and 24 post-insemination. Lactating Holstein-Friesian cows were estrous synchronized and inseminated with frozen semen. Blood samples were taken 19 and 24 days post-insemination. Serum samples were retrospectively categorized according to the pregnancy status of cows diagnosed 35 later using ultrasonography. Total RNA enriched with miRNAs was isolated from pooled (4 animals/pool) serum samples of pregnant and non-pregnant cows and subjected to cDNA synthesis. The expression of circulatory miRNAs was performed using PCR array containing primers 748 mature miRNAs. Results showed that a total of 302 and 316 miRNAs were detected in day 19 pregnant and non-pregnant cows, respectively. Similarly, 356 and 325 miRNAs were detected in day 24 pregnant and non-pregnant cows, respectively. Principal component analysis showed clear separation between pregnant and non-pregnant cows both at 19 and 24 days. We identified 8 and 23 differentially expressed miRNAs in the serum of pregnant cows of day 19 and 24, respectively. Interestingly, miR-433 and 4 other miRNAs (miR-487b, miR-495-3p, miR-376b-3p, and miR-323a-3p), which are homologous to the human pregnancy-associated C14MC miRNAs were among the differentially expressed miRNAs in day 19 and 24 pregnant cows, respectively. The adherens junction and ECM-interaction are among the pathways significantly enriched by predicted target genes of differentially expressed miRNAs. In conclusion, the expression of circulatory miRNAs in maternal blood serum of pregnant and non-pregnant cows showed distinct expression pattern and could suggest their potential involvement in early pregnancy establishment.

Differential expression of interferon-gamma, IL-4 and IL-10 in peripheral blood mononuclear cells during early pregnancy of the bovine

Maternal systemic immune response is regulated by conceptus-derived signals through peripheral blood mononuclear cells (PBMCs) via blood circulation during early pregnancy in cattle. In this study, the PBMCs from day 18 in non-pregnant cows and days 14, 18 and 30 in pregnant cows were used to explore the expression of interferon-gamma (IFN-γ), interleukin 4 (IL-4) and IL-10, and the plasma progesterone (P4) concentration was also determined. The results showed that the expression levels of mRNA and the protein of IFN-γ were lower and that IL-4 and IL-10 were higher in the PBMCs from the pregnant cows than in those of non-pregnant cows. From this study, early pregnancy induced a lower Th1 immunity (IFN-γ) and a higher Th2 immunity (IL-4 and IL-10) in the PBMCs, which may be related to interferon-tau and P4, thereby contributing to successful pregnancy in cattle.

The newly established bovine endometrial gland cell line (BEGC) forms gland acini in vitro and is only IFNτ-responsive (MAPK42/44 activation) after E2 and P4-pre-incubation

IntroductionUterine glands (UG) are crucial for the establishment of ruminant pregnancy and influenced (orchestrated manner) by estrogen (E2), progesterone (P4) and interferon tau (IFNτ). In the study we established a bovine endometrial glandular cell line (BGEC) and tested its functional reactivity (signaling) to IFNτ. MethodsBGEC was characterized by light microscopy (LM), epithelial markers (ezrin, CK18) [immunofluorescence (IF)/immunohistochemistry (IHC)] and ultrastructure (TEM/SEM) (apical microvilli). In vitro formation of gland acini and transepithelial-electric-resistance (TEER) measurements (EVOM) were done. The expression of mRNA-transcripts (RT-PCR) of steroid receptors (PR, PGRMC1/2, ESR1/2) and the IFNτ-system (IFNAR1/2, IRF1, 2, 9) was checked. BEGC was stimulated with IFNτ (10 ng/ml;1000 ng/ml) (15 min) after steroid pre-treatment [10 pg/ml E2 (two days)/20 ng/ml P4 (two days)]. Activation of MAPK42/44;STAT1 was evaluated (densitometrical Western Blot). ResultsBGEC cells expressed epithelial markers and possessed apical microvilli. High TEER-values could be measured (2320–2620 ohm/cm2). The assembled BEGC acini (25 days) were similar to UG in vivo (markers/ultrastructure). All transcripts (steroid receptors/IFNτ-system) could be detected in BEGC (mRNA). MAPK42/44 were significantly activated after E2/P4 pre-treatment and IFNτ stimulation (10 ng/ml) (p < 0.05), whilst 1000 ng/ml IFNτ did not activate MAPK42/44. Neither a STAT1 (by IFNτ) nor an activation (MAPK42/44;STAT1) by IFNτ-only was observed. DiscussionBGEC retains its epithelial phenotype in culture and forms gland acini in vitro thereby confirming its glandular character. Cells were only reactive to (low) IFNτ concentrations when pre-treated with steroids thereby closely resembling implantation physiology in vivo. BEGC can be used as a bovine implantation model to study embryo-maternal communication during early pregnancy in cattle.

Analysis of Specific Proteins for the Detection of Early Pregnancy in Cattle, A Review

Analysis of Specific Proteins for the Detection of Early Pregnancy in Cattle, A Review

Effects of early pregnancy on uterine lymphocytes and endometrial expression of immune-regulatory molecules in dairy heifers.

Natural killer (NK) cells are essential for establishment of human and rodent pregnancies. The function of these and other cytotoxic T cells (CTL) is controlled by stimulatory and inhibitory signaling. A role for cytotoxic cells during early pregnancy in cattle has not been described, but regulation of their function at the fetal-maternal interface is thought to be critical for conceptus survival. The hypothesis that the relative abundance of CTL and expression of inhibitory signaling molecules is increased by the conceptus during early pregnancy was tested. The proportions of lymphoid lineage cells and expression of inhibitory signaling molecules in the endometrium during early pregnancy in dairy heifers were determined using flow cytometry, immunofluorescence, and real-time PCR on days 17 and 20 of pregnancy and day 17 of the estrous cycle. Results revealed an increased percentage of NKp46+ and CD8+ cells in the uterus of pregnant heifers. Furthermore, a large percentage of uterine immune cells coexpressed these proteins. Compared to cyclic heifers, CD45+ uterine cells from pregnant heifers exhibited greater degranulation. Endometrium from pregnant heifers had greater mRNA abundance for the inhibitory molecules, CD274 and lymphocyte activating gene 3 (LAG3), and greater cytotoxic T lymphocyte-associated protein 4 (CTLA4), molecules that can interact with receptors on antigen-presenting cells and induce lymphocyte tolerance. This study demonstrates a dynamic regulation of both cytotoxic immune cells and tolerogenic molecules during the peri-implantation period that may be required to support establishment of pregnancy and placentation.

Understanding the uterine environment in early pregnancy in cattle: How have the omics en-hanced our knowledge?

Understanding the uterine environment in early pregnancy in cattle: How have the omics en-hanced our knowledge?

Expression of genes associated with luteolysis in peripheral blood mononuclear cells during early pregnancy in cattle.

The conceptus-derived signals that initiate maternal recognition of pregnancy act primarily on the endometrium to inhibit the development of luteolysis, thus modifying the expression of genes in the corpus luteum. The involvement of peripheral blood mononuclear cells (PBMCs) in the formation of this anti-luteolytic mechanism during early pregnancy is uncertain. In this study, PBMCs from non-pregnant and early-pregnant cows were sampled to explore the expression of genes associated with luteolysis, including AKR1B1 (aldo-keto reductase family 1, member B1; a bovine prostaglandin F synthase), PTGFR (PGF2α receptor), OXT (oxytocin), PTGES (PGE synthase), PTGER1 (PGE2 receptor 1), and PGR (progesterone receptor). OXT and PTGFR transcript abundance was low in PBMCs at Day 18 in pregnant individuals. PGR and PTGER1 mRNA abundance was significantly higher at Day 30 in pregnant individuals. AKR1B1 and PTGES transcript abundance was significantly higher at Day 18 in PBMCs from non-pregnant individuals, yet AKR1B1 and PTGES protein abundance was elevated at Day 30 in pregnant individuals-although AKR1B1 dimer may be significantly higher at Day 18 in non-pregnant PBMCs. In conclusion, changes in bovine PBMC gene expression are associated with luteolysis during early pregnancy, which implicate the influence of circulating blood components in controlling luteolysis. Mol. Reprod. Dev. 83: 509-515, 2016. © 2016 Wiley Periodicals, Inc.

Circulating miRNA signatures of early pregnancy in cattle.

BackgroundLow fertility remains a leading cause of poor productivity in dairy cattle. In this context, there is significant interest in developing novel tools for accurate early diagnosis of pregnancy. MicroRNAs (miRNAs) are short RNA molecules which are critically involved in regulating gene expression during both health and disease. MiRNAs have been shown to regulate ovarian function, uterine receptivity, embryonic development and placental function. Circulating miRNAs can provide useful biomarkers of tissue function and disease; importantly, differential miRNA profiles have been linked to pregnancy and preeclampsia in humans. This study sought to establish the potential of circulating miRNAs as biomarkers of early pregnancy in cattle.ResultsWe applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from eight non-pregnant heifers on Days 0, 8 and 16 of the oestrous cycle and 11 heifers on Days 16 and 24 of pregnancy. We sequenced a total of 46 samples and generated 9.2 million miRNA reads per sample. There were no differences in miRNA read abundance between any of the pregnant and non-pregnant time-points (FDR > 0.1). As a complementary approach, we analysed sample pools (3–4 samples/pool) corresponding to Days 0, 8 and 16 of the oestrous cycle and Day 24 of pregnancy (n = 3 pools/group) using Qiagen PCR arrays. A total of 16 miRNAs were differentially expressed (FDR < 0.1) in plasma between pregnant and non-pregnant animals. RT-qPCR validation using the same plasma samples confirmed that miR-26a was differentially upregulated on Day 16 pregnant relative to non-pregnant heifers (1.7-fold; P = 0.043), whereas miR-1249 tended to be upregulated in Day 16 pregnant heifers (1.6-fold; P = 0.081). Further validation in an independent group of heifers confirmed an increase in plasma miR-26a levels during early pregnancy, which was significant only on Day 24 (2.0-fold; P = 0.027).ConclusionsThrough genome-wide analyses we have successfully profiled plasma miRNA populations associated with early pregnancy in cattle. We have identified miR-26a as a potential circulating biomarker of early pregnancy.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2529-1) contains supplementary material, which is available to authorized users.