Early diagnosis of pregnancy in cattle is an important factor determining the profitability of breeding. It allows to shorten the intercalving period, by accelerating the next insemination procedure or diagnosing the cause of infertility. Palpation of the reproductive system by rectal examination isthe method used for more than 100 years it involves palpation of the uterine horns to detect the fetal vesicle, fetus water and the embryo itself. The earliest period when a fetal bladder can be detected is the 28th day after insemination in heifers and 32-35 in multiparous. The method is fast, cheap, does not require additional equipment, and the result is immediate. The ability to carry out the proper and effective recognition with this method is not easy and requires theoretical and practical preparation. The method also involves the risk of causing fetal damage and, as a result, loss of pregnancy and deterioration of the cow’s well-being. The modernization of the method is the introduction of an ultrasound for rectal examination, which allows to increase the efficiency of diagnosis and shorten the time from insemination to the examination. Diagnosis of early pregnancy through rectal examination, although it has been used for a long time, has not lost its importance or popularity. Although new methods of diagnosing pregnancy still appear in today’s practice, they do not affect its importance.

The molecular mechanism underlying embryonic implantation is vital to understand the correct communications between endometrium and developing conceptus during early stages of pregnancy. This study’s objective was to determine molecular changes in the uterine endometrial proteome during the preimplantation and peri-implantation between 9 days (9D), 12 days (12D), and 16 days (16D) of pregnant Polish Large White (PLW) gilts. 2DE-MALDI-TOF/TOF and ClueGOTM approaches were employed to analyse the biological networks and molecular changes in porcine endometrial proteome during maternal recognition of pregnancy. A total of sixteen differentially expressed proteins (DEPs) were identified using 2-DE gels and MALDI-TOF/TOF mass spectrometry. Comparison between 9D and 12D of pregnancy identified APOA1, CAPZB, LDHB, CCT5, ANXA4, CFB, TTR upregulated DEPs, and ANXA5, SMS downregulated DEPs. Comparison between 9D and 16D of pregnancy identified HP, APOA1, ACTB, CCT5, ANXA4, CFB upregulated DEPs and ANXA5, SMS, LDHB, ACTR3, HP, ENO3, OAT downregulated DEPs. However, a comparison between 12D and 16D of pregnancy identified HP, ACTB upregulated DEPs, and CRYM, ANXA4, ANXA5, CAPZB, LDHB, ACTR3, CCT5, ENO3, OAT, TTR down-regulated DEPs. Outcomes of this study revealed key proteins and their interactions with metabolic pathways involved in the recognition and establishment of early pregnancy in PLW gilts.

During the early stages of pregnancy, the uterine endometrium undergoes dramatic morphologic and functional changes accompanied with dynamic variation in gene expression. Pregnancy-stage specific differentially expressed gene (DEG)-transcript-probes were investigated and identified by comparing endometrium transcriptome at 9th day (9D), 12th day (12D) and 16th day (16D) of early pregnancy in Polish large-white (PLW) gilts. Endometrium comparisons between 9D-vs-12D, 9D-vs-16D and 12D-vs-16D of early pregnancy identified 6049, 374 and 6034 highly significant DEG-transcript-probes (p < 0.001; >2 FC). GO term enrichment analysis identified commonly shared upregulated endometrial DEG-transcript-probes (p < 0.001; >2 FC), that were regulating the gene functions of anatomic structure development and transport (TG), DNA-binding and methyltransferase activity (ZBTB2), ion-binding and kinase activity (CKM), cell proliferation and apoptosis activity (IL1B). Downregulated DEG-transcript-probes (p < 0.001; >2 FC) were involved in regulating the gene functions of phosphatase activity (PTPN11), TC616413 gene-transcript and Sus-scrofa LOC100525539. Moreover, blastn comparison of microarray-probes sequences against sus-scrofa11 assembly identified commonly shared upregulated endometrial DEG-transcript-probes (E < 0.06; >2 FC), that were regulating the gene functions of reproduction and growth (SELENOP), cytoskeleton organization and kinase activity (CDC42BPA), phosphatase activity (MINPP1), enzyme-binding and cell-population proliferation (VAV3), cancer-susceptibility candidate gene (CASC4), cytoskeletal protein-binding (COBLL1), ion-binding, enzyme regulator activity (ACAP2) Downregulated endometrial DEG-transcript-probes (E < 0.06; >2FC) were involved in regulating the gene functions of signal-transduction (TMEM33), catabolic and metabolic processes (KLHL15). Microarray validation experiment on selected candidate genes showed complementarity to significant endometrial DEG-transcript-probes responsible for the regulation of immune response (IL1B, S100A11), lipid metabolism (FABP3, PPARG), cell-adhesion (ITGAV), angiogenesis (IL1B), intercellular transmission (NMB), cell-adhesion (OPN) and response to stimuli (RBP4) was confirmed by RT-PCR. This study provides a clue that identified pregnancy-stage specific microarray transcript probes could be considered as candidate genes for recognition and establishment of early pregnancy in the pig.

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Assisted reproductive techniques in cattle, such as artificial insemination (AI) and embryo transfer (ET), are widely used. Despite many years of methodological improvements, the pregnancy rate (PR) in cows has not increased in direct proportion with their development. Among the possibilities to increase the PR is the use of certain steroids and nonsteroidal anti-inflammatory drugs (NSAIDs). The antiluteolytic effect of NSAIDs is achieved by blocking cyclooxygenase, which is involved in the conversion of arachidonic acid to prostaglandins. This article compares the PRs obtained after treatment with the commonly used NSAIDs in cattle, including flunixin meglumine, carprofen, meloxicam, ibuprofen, aspirin, and sildenafil. Studies on the effectiveness of certain steroid drugs on the PR have also been described. The results were not always consistent, and so comparisons between studies were made. In conclusion, flunixin meglumine seems to be an option, and can be recommended for improving ET results, especially in situations of high exposure or susceptibility to stress. Its administration under all circumstances, however, might be pointless and will not lead to the desired effect.