Early pregnancy; Pregnancy-associated glycoprotein; Progesterone; Radioimmunoassay; Sarda ewe
Abstract :
[en] The aim of the current study was to describe the use of a pool of different antisera raised against pregnancy-associated glycoproteins (PAGs; purified from both ovine and caprine placentas) for early pregnancy diagnosis in ovine species. Sixty-three pluriparous Sarda ewes (Ovis aries) were synchronized. Blood samples were withdrawn on Days 18, 24, 26, 28, 30, and 50 after mating. These samples were assayed for progesterone (radioimmunoassay [RIA] including an extraction step) and for pregnancy-associated glycoproteins (RIA-706 and RIA-srPool). Progesterone concentrations were under 1.0ng/mL in all nonpregnant Sarda ewes. In pregnant ewes, mean progesterone concentrations ranged from 2.4ng/mL (Day 24, single pregnancies) to 4.4ng/mL (Day 28, multiple pregnancies). During all periods of examination, PAGs remained lower than 0.8ng/mL in nonpregnant ewes. On Day 18 of pregnancy, PAG concentrations could be detected in 26 of 43 (60.5%) and in 41 of 43 (95.3%) pregnant ewes using the RIA-706 and RIA-srPool methods, respectively. From Day 24 to Day 50, using both RIA methods, PAGs could be detected in all pregnant ewes. On Day 24, the best threshold for pregnancy diagnosis was obtained by use of RIA-srPool, maximal concentration in nonpregnant ewes being 0.3ng/mL and minimal concentration in pregnant ewes being 4.8ng/mL. In general, progesterone and PAG concentrations were higher in multiple pregnancies than in single pregnancies. However, because of large individual variations, single pregnancies could not be differentiated from multiple pregnancies
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Barbato, Olimpia; University of Perugia Italy
Melo de Sousa, Noelita ; Université de Liège - ULiège > Département de sciences fonctionnelles > Physiologie de la reproduction
Debenedetti, A.; University of Perugia Italy
Canali, C.; University of Perugia Italy
Todini, L.; University of Camerino Italy
Beckers, Jean-François ; Université de Liège - ULiège > Département de sciences fonctionnelles > Physiologie de la reproduction
Language :
English
Title :
Validation of a new pregnancy-associated glycoprotein radioimmunoassay method for the detection of early pregnancy in ewes.
Maciotta N.P.P., Cappio-Borlino A., and Pulina G. Analysis of environmental effects on tests day milk yields of Sarda dairy ewes. J Dairy Sci 82 (1999) 2212-2217
Boscos C.M., Samartzi F.C., Lymberopoulos A.G., Stefanakis A., and Belibasaki S. Assessment of progesterone concentration using enzyme immunoassay for early pregnancy diagnosis in sheep and goats. Reprod Domest Anim 38 (2003) 170-174
Sasser R.C., and Ruder C.A. Detection of early pregnancy in domestic ruminants. J Reprod Fertil 34 Suppl (1987) 261-271
McPhee I.M., and Tiberghien M.P. Assessment of pregnancy in sheep by analysis of plasma progesterone using amplified enzyme immunoassay technique. Vet Rec 121 (1987) 63-65
Susmel P., and Piasentier E. Assessment of pregnancy in Bergamasca ewes by analysis of plasma progesterone. Small Rumin Res 8 (1992) 325-332
Gvozdic D., and Ivkov V. Early pregnancy diagnosis in ewes. Acta Vet Beograde 44 (1994) 215-219
Kalkan C., Cetin H., Kaygusuzoglu E., Yilmaz B., Ciftci M., Yidiz H., Yildiz A., Deveci H., et al. An investigation on plasma progesterone levels during pregnancy and parturition in the Ivesi sheep. Acta Vet Hung 44 (1996) 335-340
Ranilla M.J., Sulon J., Carro M.D., Mantecon A.R., and Beckers J.F. Plasmatic profiles of pregnancy-associated glycoprotein and progesterone levels during gestation in Churra and Merino sheep. Theriogenology 42 (1994) 537-545
Willard J.M., White D.R., Wesson C.A.R., Stellflug J., and Sasser R.G. Detection of fetal twins in sheep using a radioimmunoassay for pregnancy-specific protein B. J Anim Sci 73 (1995) 960-966
Ranilla M.J., Sulon J., Mantecon A.R., Beckers J.F., and Carro M.F. Plasma pregnancy-associated glycoprotein and progesterone concentrations in pregnant Assaf ewes carrying single and twin lambs. Small Rumin Res 24 (1997) 125-131
Karen A., El Amiri B., Beckers J.F., Sulon J., Taverne M.A.M., and Szenzi O. Comparison of accuracy of transabdominal ultrasonography, progesterone and pregnancy-associated glycoproteins tests for discrimination between single or multiple pregnancy in sheep. Theriogenology 66 (2006) 314-322
Ledezma-Torres R.A., Beckers J.F., and Holtz W. Assessment of plasma profile of pregnancy-associated glycoprotein (PAG) in sheep with a heterologous (anti-caPAG22 + 59) RIA and its potential for diagnosing pregnancy. Theriogenology 66 (2006) 906-912
Vandaele L., Verberckmoes S., El Amiri B., Sulon J., Duchateau L., Van Soom A., et al. Use of homolougous radioimmunoassay (RIA) to evaluate the effect of maternal and foetal parameters on pregnancy-associated glycoprotein (PAG) concentrations in sheep. Theriogenology 63 (2005) 1914-1924
El Amiri B., Sulon J., Karen A., Alvarez A.V., Cogniè Y., Sousa N.M., et al. Measurement of ovine pregnancy-associated glycoprotein (PAG) during early pregnancy in Lacaune sheep. Reprod Domest Anim 42 (2007) 257-262
Xie S., Green J.A., Bao B., Beckers J.F., Valdez K.E., Hakami L., and Roberts R.M. Multiple pregnancy-associated glycoproteins are secreted by day 100 ovine placental tissue. Biol Reprod 57 (1997) 1384-1393
Xie S., Green J., Bixby J.B., Szafranska B., DeMartini J.C., Hecht S., and Roberts R.M. The diversity and evolutionary relationships of the pregnancy-associated glycoproteins, an aspartic proteinase subfamily consisting of many trophoblast-expressed genes. Proc Natl Acad Sci U S A 94 (1997) 12809-12816
Green JA, Xie S, Quan X, Bao B, Gan X, Mathialagan N, Roberts RM. Pregnancy-associated glycoproteins exhibit spatially and temporally distinct expression patterns during pregnancy. Biol Reprod 200, 62, 1624-1632.
Garbayo J.M., Green J.A., Manikkam M., Beckers J.F., Kiesling D.O., Ealy A.D., and Roberts R.M. Caprine pregnancy-associated glycoproteins (PAG): their cloning, expression, and evolutionary relationship to other PAG. Mol Reprod Dev 57 (2000) 311-322
Boiti C., Ceccarelli P., Beghelli V., Daniotti P., and Pennisi F. A RIA method for plasma progesterone determination. Proc Soc Ital Sci Vet 26 (1974) 366-371
Todini L., Malfatti A., Barbato O., Costarelli S., and Debenedetti A. Progesterone plus PMSG priming in seasonally anovulatory lactating Sarda ewes exposed to the ram effect. J Reprod Dev 53 (2007) 437-441
Zoli A.P., Beckers J.F., Wouters-Ballman P., Closset J., Falmagne P., and Ectors F. Purification and characterization of a bovine pregnancy-associated glycoproteins. Biol Reprod 45 (1991) 1-10
Zoli A.P., Guilbault L.A., Delahaut P., Ortiz W.B., and Beckers J.F. Radioimmunoassay of a bovine pregnancy-associated glycoprotein in serum: its application for pregnancy diagnosis. Biol Reprod 46 (1992) 83-92
Greenwood F.C., Hunter W.M., and Glover J.S. The preparation of 131-I labeled human growth hormone of high specific radioactivity. Biochemistry 89 (1963) 114-123
Perenyi Z., Szenci O., Sulon J., Drion P.V., and Beckers J.F. Comparison of the ability of three radioimmunoassays to detect pregnancy-associated glycoproteins in bovine plasma. Reprod Domest Anim 37 (2002) 100-104
Perenyi Z., Szenci O., Drion P.V., Banga-Mboko H., Sousa N.M., El Amiri B., and Beckers J.F. Aspartic proteinase members secreted by the ruminant placenta: specificity of three radioimmunoassay systems for the measurement of pregnancy-associated glycoproteins. Reprod Domest Anim 37 (2002) 324-329
Garbayo J.M., Remy B., Alabart J.L., Folch J., Wattiez R., Falmagne P., and Beckers J.F. Isolation and partial characterization of a pregnancy-associated glycoprotein family from the goat placenta. Biol Reprod 58 (1998) 109-115
El Amiri B., Remy B., Sousa N.M., Joris B., Otthiers N.G., Perényi Z., et al. Isolation and partial characterization of three pregnancy-associated glycoproteins from the ewe placenta. Mol Reprod Dev 64 (2003) 199-206
El Amiri B., Remy B., Sousa N.M., and Beckers J.F. Isolation and characterization of eight pregnancy-associated glycoproteins present at high levels in the ovine placenta between day 60 and day 100 of gestation. Reprod Nutr Dev 44 (2004) 169-181
Vaitukaitis J., Robbins J.B., Nieschlag E., and Ross G.T. A method for producing specific antisera with small doses of immunogen. J Clin Endocrinol Metab 33 (1971) 988-991
Ayad A., Sousa N.M., Sulon J., Iguer-Ouada M., and Beckers J.F. Comparison of five radioimmunoassay systems for PAG measurement: ability to detect early pregnancy in cows. Reprod Domest Anim 42 (2007) 433-440
Szenci O., Beckers J.F., Sulon J., Sasser G., Taverne M.A.M., Varga J., et al. Comparison of ultrasonography, bovine pregnancy specific B, and bovine pregnancy-associated glycoprotein 1 test for pregnancy detection in dairy cows. Theriogenology 50 (1998) 77-88
Wallace J.M., Aitken R.P., Cheyne M.A., and Humblot P. Pregnancy-specific protein B and progesterone concentrations in relation to nutritional regimen, placental mass and pregnancy outcome in growing adolescent ewes carrying singleton fetuses. J Reprod Fertil 109 (1997) 53-58
Karen A., Beckers J.F., Sulon J., Melo de Sousa N., Szbados K., Reczigel J., and Szenzi O. Early pregnancy diagnosis in sheep by progesterone and pregnancy-associated glycoprotein tests. Theriogenology 59 (2003) 1941-1948
Karen A., Beckers J.F., Sulon J., el Amiri B., Szabados K., Ismail S., et al. Evaluation of false transrectal ultrasonographic pregnancy diagnosis in sheep by measuring the plasma level of pregnancy-associated glycoproteins. Reprod Nutr Dev 43 (2003) 577-586
Patel O.V., Sulon J., Beckers J.F., Takahashi T., Hirako M., Sasaki N., and Domeki I. Plasma bovine pregnancy-assaciated glycoprotein concentrations throughout gestation in relationship to fetal number in the cow. Eur J Endocrinol 137 (1997) 423-428
Mukasa-Mugerwa E., and Viviani P. Progesterone concentrations in peripheral plasma of Menz sheep during gestation and parturition. Small Rumin Res 8 (1992) 47-53
Quirke J.F., Hanrahan J.P., and Gosling J.P. Plasma progesterone levels throughout the oestrus cycle and release of LH at oestrus in sheep with different ovulation rate. J Reprod Fertil 55 (1979) 37-44
