Early Pregnancy Diagnosis in Bovines: Current Status and Future Directions, Central Institute for Research on Buffaloes, Hisar 125001, India, LLR University of Veterinary and Animal Sciences, Hisar 125005, India, National Dairy Research Institute, Karnal 132001, India. Currently available direct methods for diagnosis of pregnancy include transrectal palpation and B-mode ultrasonography, and both methods are currently used by bovine practitioners to diagnose pregnan… Further purification and characterization of several isoforms from bovine foetal cotyledons found that protein B is actually a 67 kDa PAG [91]. Green, T. E. Parks, M. P. Avalle et al., “The establishment of an ELISA for the detection of pregnancy-associated glycoproteins (PAGs) in the serum of pregnant cows and heifers,”, E. Silva, R. A. Sterry, D. Kolb et al., “Accuracy of a pregnancy-associated glycoprotein ELISA to determine pregnancy status of lactating dairy cows twenty-seven days after timed artificial insemination,”, A. Ayad, N. M. Sousa, J. Sulon, J. L. Hornick, M. Iguer-Ouada, and J. F. Beckers, “Correlation of five radioimmunoassay systems for measurement of bovine plasma pregnancy-associated glycoprotein concentrations at early pregnancy period,”, N. M. de Sousa, M. Zongo, W. Pitala et al., “Pregnancy-associated glycoprotein concentrations during pregnancy and the postpartum period in, A. Karen, S. Darwish, A. Ramoun et al., “Accuracy of ultrasonography and pregnancy-associated glycoprotein test for pregnancy diagnosis in buffaloes,”, N. M. Sousa, A. Ayad, J. F. Beckers, and Z. Gajewski, “Pregnancy-associated glycoproteins (PAG) as pregnancy markers in the ruminants,”, P. R. Graves and T. A. J. Haystead, “Molecular biologist's guide to proteomics,”, J. D. Lippolis and T. A. Reinhardt, “Centennial paper: proteomics in animal science,”, D. A. Colantonio and D. W. Chan, “The clinical application of proteomics,”, F. di Girolamo, E. Boschetti, M. C. M. Chung, F. Guadagni, and P. G. Righetti, “‘Proteomineering’ or not? It can be assayed in milk or plasma. Several methods of pregnancy diagnosis are being practiced in bovine species, yet none qualifies as the ideal pregnancy diagnosis method due to the inherent limitations of sensitivity, accuracy, specificity, speed, and ease of performing the test. Early Pregnancy Diagnosis and Its Importance in Dairy Cattle Management – epashupalan Early pregnancy diagnosis is essential to maintain the farm in a profitable manner. Concentration of estrone sulphate in the maternal body fluids is a useful indicator for the placental functions especially those related to embryonic growth [44]. Early pregnancy factor (EPF, also known as early conception factor—ECF)—a 10.84 kDa protein [50], is present in the sera of pregnant mammalian females, detectable within 6 to 24 hours of fertilization [51] and disappearing within 24 to 48 after death or removal of the embryo [52], EPF is present in the serum up to two-thirds of the gestation [53]. These findings support other findings in cattle which claim that transrectal ultrasonography for pregnancy diagnosis between days 21 and 25 after breeding has sensitivity and specificity of 44.8% and 82.3%, respectively, which further increase to 97.7% and 87.7%, respectively, when conducted between 26 and 33 days after AI [18]. Early identification of non-pregnant cows post breeding improves reproductive efficiency and pregnancy rate in cattle by decreasing the interval between AI services and increasing AI service rate. Therefore early pregnancy diagnosis prior to 21 days post insemination may be unreliable. Both require a great deal of skill and experience. In conclusion, early pregnancy diagnosis can provide opportunities to improve profitability. Pregnancy diagnosis in dairy cattle is traditionally conducted by rectal palpation or transrectal ultrasonography. The progesterone secreted by a functional corpus luteum between 18 and 24 days after service or insemination is an early indication of pregnancy. An early and precise pregnancy diagnosis is an important criterionforbetterreproductivemanagementinlivestocklike cows and bu aloes. Early pregnancy diagnosis is an important aspect for optimizing dairy production, yet none of the present day methods qualifies as an ideal diagnostic due to limitations of accuracy, later stages of applicability, and requirement for elaborate instrumentation and laboratory setup. Conception extends the life of the corpus luteum (CL) by preventing the luteolytic mechanism from being triggered, thus prolonging and maintaining its functional characteristics, ensuring continued high progesterone levels [31]. Pregnancy loss in beef cattle causes both management and economic challenges to a producer. By May 2010, already there were 2 million cattle blood tests conducted for pregnancy detection (http://biotrackingcom.siteprotect.net/about/timeline). Data generated out of sequential blood proteome analysis during pregnancy can have several other applications as well, for example, studying fetal viability, genetic disorders, and so forth. Moor and Rowson [69], the pioneers of sheep embryo transfer, transferred embryos on days 12, 13, and 14 to unmated ewes and suggested interactions between the embryo and uterus that influence the luteal function and result in establishment of pregnancy. Because of the high incidence of embryonic mortality this test should be treated solely as an indication of conception. Recently available tests detect so called early conception factor (ECF) or pregnancy-associated glycoprotein in blood samples. A. abnormal uterine contents (pyometra or mucometra), incorrect handling of the samples and test kit. expressed over a considerable period of time to give ample time for diagnosis, http://biotrackingcom.siteprotect.net/about/timeline, Fertilization, single-cell embryo (zygote) in oviduct, Early cleavages in the oviduct (up to 8 cell stages), activation of embryonic genome, 16–32 cell zona-enclosed embryo progressing into compact morula stage, Formation of a blastocoele with differentiation of embryonic cells, Blastocyst expansion and hatching from the zona pellucid, Blastocyst elongation from a tubular to a filamentous structure, Compiled from available information (Morris and Diskin, 2008 [, “Committee on Reproductive Nomenclature Recommendations for standardizing bovine reproductive terms,”, N. Ayalon, “A review of embryonic mortality in cattle,”, D. Morris and M. Diskin, “Effect of progesterone on embryo survival,”, W. Wisnicky and L. E. Cassida, “A manual method for diagnosis of pregnancy in cattle,”, G. H. Arthur, D. E. Noakes, H. Pearson, and T. J. Parkinson, “Reproduction in the buffalo,” in, O. J. Franco, M. Drost, M.-J. PAGs are used for development of bench-top pregnancy detection methods [110], which are now commercially available as BioPRYN (BioTracking, Russia), DG29 (Genex Cooperative Inc., USA), and IDEXX (IDEXX Laboratories, Inc., USA). In the absence of a single indicator for a particular life process, a combination of the expression patterns of more than one substance can be used for a purposeful analysis as in the quadruple test where levels of four blood constituents (alpha-fetoprotein, human chorionic gonadotropin, unconjugated oestriol, and inhibin-A) have been used to predict the probability of Down’s syndrome in babies [119]. Table 2 describes the work in different labs on the level of progesterone in pregnant and nonpregnant bovines. It has some risks associated with it in terms of potentially causing early embryonic loss. accuracy in detecting pregnancy) of the cow-side milk progesterone (EIA) test was 93.1% in a study by Pieterse et al. The very fact that pregnancy brings about numerous physiological changes in the female body through secretion/altered secretion of various biomolecules, which often are proteins or their metabolites, supports research endeavours aimed at identifying novel proteins as the candidate molecules for pregnancy detection. The early pregnancy diagnosis allows optimizing production and timely management correction, with a greater reproductive output of livestock. Delaying in identifying the non pregnant animal leads to economic loss to the farmers. present in easily accessible body fluids like serum, milk, urine, and vaginal discharge. Transrectal ultrasonography has the added advantage of providing additional information on ovarian structures, identification of twins, and determination of fetal viability, age, and sex [10, 12]. A chemical test for pregnancy termed Early Pregnancy Factor (EPF) or Early Conception Factor (ECF) was proposed and marketed in the 1990’s (Concepto Diagnostics, Knoxville,TN) This molecule was supposedly present in the blood of pregnant cattle within two days after conception The exact nature of … It has some risks associated with it in terms of potentially causing early embryonic … A pregnancy detection test on the same lines as the human quadruple test can be tried where instead of one we can consider protein profiles of more proteins. Copyright © 2013 Ashok K. Balhara et al. Early detection of pregnancy is a benefit to producers, but it is important to remember most early embryonic mortality (pregnancy loss) occurs prior to day 60. accuracy in detecting non-pregnancy) was only 39.3%. Interferon- These included nine pregnancy-specific spots in day 21 and day 35 serum samples. Many methods of pregnancy diagnosis, both direct and indirect, are being practiced in bovine species; none till date actually qualifies as the ideal pregnancy diagnosis method due to the limitations they inherit. stimulated, are upregulated during early pregnancy [84–86]. Several methods are available to determine if a cow or heifer is pregnant. Of these, the former showed an immune reaction similar to that of Interferon- Rectal palpation is widely used and considered a safe method for pregnancy diagnosis in cattle. Pregnancy should be confirmed later by rectal or ultrasound examination. PAGs are a very complex group of proteins, a fact proven by the already documented 22 distinct cDNA libraries [94]. Currently available state-of-the-art instrumentation and proteomics techniques instil hope for finding molecules—exclusively related to intricate maternal metabolic alterations necessary to align with physiology of early embryonic development and its signalling for maternal recognition of pregnancy and continued survival. This may suggest that these proteins could be pregnancy-specific subunits or fragments of albumin and IgG or carrying differentially expressed small proteins, which may ultimately have potential for pregnancy detection. Nonetheless early or inappropriate palpation of the amniotic vesicle may damage the embryo and cause embryonic mortality. Accuracy Early pregnancy diagnosis in cattle. , a 172 amino acid polypeptide [73], blocks transcription of estrogen receptor alpha and oxytocin receptors in endometrial cells [78], while downregulating the expression of enzymes cyclooxygenase-2 and prostaglandin F synthase [79], thus preventing PGF release necessary for luteolysis. Encouragingly enough, these studies need further investigations for arriving at some sort of pregnancy detection method. Additionally, the biological marker for pregnancy should have the following desired characteristics:(i)specifically upregulated or downregulated during pregnancy,(ii)least affected by nonanimal factors like feed, environment, and drug interactions,(iii)having the ability to reflect age as well as viability of the conceptus,(iv)present in easily accessible body fluids like serum, milk, urine, and vaginal discharge,(v)expressed over a considerable period of time to give ample time for diagnosis,(vi)revealing the result immediately. However, even if oestrus detection is good, not all of these cows will be pregnant. having the ability to reflect age as well as viability of the conceptus. All these experiments have suggested IFN- specifically upregulated or downregulated during pregnancy. Its concentrations increase from day 60 and plateau around day 150 after insemination [42]. vesicle, Common reasons for errors in rectal palpation, Safety King, “Conjugated and unconjugated oestrogens in fetal and maternal fluids of the cow throughout pregnancy,”, R. B. Heap and M. Hamon, “Oestrone sulphate in milk as an indicator of a viable conceptus in cows,”, M. Hamon, I. R. Fleet, R. J. Holdsworth, and R. B. Heap, “The time of detection of oestrone sulphate in milk and the diagnosis of pregnancy in cows,”, M. Hirako, T. Takahashi, and I. Domeki, “Peripheral changes in estrone sulfate concentration during the first trimester of gestation in cattle: comparison with unconjugated estrogens and relationship to fetal number,”, B. S. Prakash and M. L. Madan, “Influence of gestation on oestrone sulphate concentration in milk of zebu and crossbred cows and murrah buffaloes,”, F. Lobago, M. Bekana, H. Gustafsson et al., “Serum profiles of pregnancy-associated glycoprotein, oestrone sulphate and progesterone during gestation and some factors influencing the profiles in ethiopian borana and crossbred cattle,”, S. Aschheim and B. Zondek, “Hypophysenvorderlappenhormon und Ovarialhormon im Harn von Schwangeren,”, J. R. Marshall, C. B. Hammond, G. T. Ross, A. Jacobson, P. Rayford, and W. D. Odell, “Plasma and urinary chorionic gonadotropin during early human pregnancy,”, K. Imakawa, R. V. Anthony, M. Kazemi, K. R. Marotti, H. G. Polites, and R. M. Roberts, “Interferon-like sequence of ovine trophoblast protein secreted by embryonic trophectoderm,”, A. C. Cavanagh, “Identification of early pregnancy factor as chaperonin 10: implications for understanding its role,”, H. Morton, V. Hegh, and G. J. Reliable methods for pregnancy diagnosis is an important criterion for better reproductive management in livestock cows to be least... 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