Altered secretion of pregnancy-associated glycoproteins during gestation in bovine somatic clones.
; ; et al
in Theriogenology (2011), 76(6), 1006-1021
Somatic nuclear transfer (NT) in cattle is often accompanied by severe placental anomalies, hypertrophy, and hydrallantois, which induce a high rate of pregnancy losses throughout gestation. These ... [more ▼]
Somatic nuclear transfer (NT) in cattle is often accompanied by severe placental anomalies, hypertrophy, and hydrallantois, which induce a high rate of pregnancy losses throughout gestation. These placental deficits are associated with an abnormal increase of the maternal plasma levels of pregnancy-associated glycoprotein (PAG), produced by the trophoblastic binucleate cells (BNC) of the placenta. The objective of this study was to analyze the origin of the abnormally elevated PAG concentrations in the peripheral circulation of NT recipients during pathological pregnancies. Concentrations of PAG were measured both in maternal blood, in chorionic and cotyledonary tissular extracts from control recipients (after artificial insemination, AI, or in vitro fertilization, IVF) and clone recipients on Day 32, Day 62, and during the third trimester of gestation. Three different radioimmunoassay (RIA) systems were used. One homologous RIA for PSP60, similar to bovine PAG-1 (PAG(67kDa)), and two heterologous RIA with PAG(67kDa) as standard and tracer, and antisera anti-caprine PAG (AS#706 and AS#708). Circulating and tissular concentrations of bovine placental lactogen (bPL), a glycoprotein also produced by BNC, were determined by RIA at the same stages. The number of BNC in the placental tissues was determined by cell counting after immunostaining with anti PSP60 antibody on tissue sections from control and NT pregnancies. Maternal plasma PAG concentrations were not different among groups on Day 32, but they were significantly higher in NT than in control pregnancies on Day 62 with all three RIA and during the third trimester with two RIA (RIA-PSP60 and RIA with AS#708). Circulating bPL concentrations were undetectable on Days 32 and 62 and were not different in the third trimester between NT and control pregnancies. Tissular amounts of PAG on total proteins were not different between the two groups at all stages studied. No difference was determined in the percentage of PSP60-positive BNC in placental tissues between controls and NT on Day 62 and during the third trimester of pregnancy. Western blots of tissular extracts from placenta showed no major molecular weight changes of PAG in NT pregnancies compared to controls. No differences in maternal circulation concentrations or tissular content of bPL were observed between control and NT pregnancies. In conclusion, the specific increase of PAG in maternal plasma concentrations during abnormal NT pregnancies do not result from a higher proportion of BNC, or an increased protein expression of PAG and could be due to changes in the composition of terminal glycosylation which result into a clearance decrease of PAG from the circulation. [less ▲]Detailed reference viewed: 30 (6 ULg)
Sex and PRNP genotype determination in preimplantation caprine embryos
; ; et al
in Reproduction in Domestic Animals (2011), 46(4), 656-663
The objective of this study was to test the accuracy of genotype diagnosis after whole amplification of DNA extracted from biopsies obtained by trimming goat embryos and to evaluate the viability of ... [more ▼]
The objective of this study was to test the accuracy of genotype diagnosis after whole amplification of DNA extracted from biopsies obtained by trimming goat embryos and to evaluate the viability of biopsied embryos after vitrification/warming and transfer. Whole genome amplification (WGA) was performed using Multiple Displacement Amplification (MDA). Sex and prion protein (PRNP) genotypes were determined. Sex diagnosis was carried out by PCR amplification of ZFX/ZFY and Y chromosome-specific sequences. Prion protein genotype determination was performed on codons 142, 154, 211, 222 and 240. Embryos were collected at day 7 after oestrus and biopsied either immediately after collection (blastocysts and expanded blastocysts) or after 24 h of in vitro culture (compacted morulae). Biopsied embryos were frozen by vitrification. Vitrified whole embryos were kept as control. DNA of biopsies was extracted and amplified using MDA. Sex diagnosis was efficient for 97.4% of biopsies and PRNP genotyping was determined in 78.7% of biopsies. After embryo transfer, no significant difference was observed in kidding rate between biopsied and vitrified control embryos, whereas embryo survival rate was different between biopsied and whole vitrified embryos (p = 0.032). At birth, 100% of diagnosed sex and 98.2% of predetermined codons were correct. Offspring PRNP profiles were in agreement with parental genotype. Whole genome amplification with MDA kit coupled with sex diagnosis and PRNP genotype predetermination are very accurate techniques to genotype goat embryos before transfer. These novel results allow us to plan selection of scrapie-resistant genotypes and kid sex before transfer of cryopreserved embryo. [less ▲]Detailed reference viewed: 44 (3 ULg)
The Efficiency of In vitro Ovine Embryo Production Using an Undefined or a Defined Maturation Medium is Determined by the Source of the Oocyte.
; ; et al
in Reproduction in Domestic Animals (2011), 46(3), 463-470
In vitro oocyte maturation can be influenced by oocyte source and maturation media composition. The aim of the present study was to compare the efficiency of a defined in vitro maturation medium (TCM199 ... [more ▼]
In vitro oocyte maturation can be influenced by oocyte source and maturation media composition. The aim of the present study was to compare the efficiency of a defined in vitro maturation medium (TCM199 supplemented with cysteamine and epidermal growth factor; Cys + EGF) with an undefined medium (TCM199 supplemented with follicle-stimulating hormone and follicular fluid; FSH + FF) for in vitro production (IVP) of ovine embryos, using oocytes obtained by laparoscopic ovum pick-up from FSH-stimulated [n = 11; 158 cumulus-oocyte complexes (COCs)] and non-stimulated (n = 16; 120 COCs) live ewes, as well as abattoir-derived oocytes (170 COCs). The produced blastocysts were vitrified and some of them were transferred to synchronized recipients. The best and the worst final yields of embryo IVP observed in this study were obtained using oocytes from FSH-stimulated ewes matured in FSH + FF (41.3%; 33/80) and in Cys + EGF (19.2%; 15/78) medium, respectively (p < 0.01). No significant differences between both media were attained in the blastocyst development rate or in the final yield of embryo IVP using oocytes from non-stimulated ewes or abattoir-derived oocytes. The overall in vivo survival rate of the transferred vitrified blastocysts was 13.1% (8/61), without significant differences between oocyte sources or maturation media. In conclusion, under the experimental conditions of the present study, TCM199 supplemented with cysteamine and EGF is a convenient defined maturation medium for IVP of embryos from oocytes of live non-stimulated ewes or from oocytes of abattoir-derived ovaries. However, the best final yield of embryo IVP observed in this study was attained when oocytes came from FSH-stimulated donors and TCM199 was supplemented with FSH and follicular fluid [less ▲]Detailed reference viewed: 23 (1 ULg)
Accuracy of transrectal palpation for early pregnancy diagnosis in Egyptian buffaloes.
; ; et al
in Tropical Animal Health & Production (2011), 43(1), 5-7
The present study was undertaken to evaluate the accuracy of transrectal palpation (TRP) for diagnosing early pregnancy in buffaloes and the false diagnoses of the TRP test by using the pregnancy ... [more ▼]
The present study was undertaken to evaluate the accuracy of transrectal palpation (TRP) for diagnosing early pregnancy in buffaloes and the false diagnoses of the TRP test by using the pregnancy-associated glycoprotein radioimmunoassay (PAG-RIA) test. Pregnancy was diagnosed in 168 buffalo-cows once by TRP and PAG-RIA test between days 31 and 55 after breeding. The sensitivity of TRP for detecting pregnant buffalo-cows was 37.5% at days 31-35, increased to 93.8% at days 46-50 and reached 100% at days 51-55 (P < 0.01). All cases of false negative diagnoses (n = 10) had PAG concentration higher than the threshold (>/=1.8 ng/mL) for diagnosing pregnancy. The specificity of TRP for detecting non-pregnant buffalo cows ranged between 90.9%, and 100% between days 31 and 55. All cases of false positive diagnoses (n = 5) made by TRP had PAG concentrations lower than the threshold for diagnosing pregnancy. It could be concluded that TRP is an accurate method for diagnosing pregnant and non-pregnant buffalo cows from day 46 after breeding. [less ▲]Detailed reference viewed: 56 (3 ULg)
The pregnancy-associated glycoproteins. Clinical applications for pregnancy follow-up in ruminant species.
; Melo de Sousa, Noelita ; et al
in Revista română de medicină veterinară (2011), 21(2), 117-126
Pregnancy diagnosis is an important part in reproduction management of ruminant and embryonic mortality has a substantial impact on the fertility of demestic animals. This article focus on the use of ... [more ▼]
Pregnancy diagnosis is an important part in reproduction management of ruminant and embryonic mortality has a substantial impact on the fertility of demestic animals. This article focus on the use of pregnancy-associated glycoproteins for pregnancy follow-up. [less ▲]Detailed reference viewed: 25 (1 ULg)
Plasma urea nitrogen in relation to pregnancy rate in dairy sheep
; Beckers, Jean-François ; Melo de Sousa, Noelita et al
in Animal reproduction science (2011), 124(1-2), 69-72
The aim of this field study was to investigate the relationship of plasma urea nitrogen (PUN) with the pregnancy rate in lactating AwassixMerino ewes. One hundred and eighty-five AwassixMerino ewes were ... [more ▼]
The aim of this field study was to investigate the relationship of plasma urea nitrogen (PUN) with the pregnancy rate in lactating AwassixMerino ewes. One hundred and eighty-five AwassixMerino ewes were used in the present study. Ewes were fed a diet containing 17.4% crude protein and were milked twice a day by the milking machine. The ewes were synchronized for estrus by insertion of intravaginal sponges containing 30mg flurogestone acetate for 14 days. At the time of sponge removal each ewe was administered eCG (600IU). All ewes were inseminated twice with fresh semen into the external os of the cervix at 48 and 56h after sponge removal. The day of insemination was considered as Day 0 for calculating the gestational period. Blood samples were collected from each ewe at Days 0, 18 for measurement of PUN concentrations and at Day 22 after AI for measurement of pregnancy-associated glycoprotein (PAG) by radioimmunoassay (RIA). Thirty-eight ewes (20.5%) were confirmed pregnant by PAG-RIA test at Day 22 and by ultrasonography at Day 80. The mean (+/-S.D.) concentration of PUN in all ewes at Day 0 was 12.7+/-4.6mmol/L. There were non-significant differences in the level of PUN between pregnant and non-pregnant ewes at Days 0 (12.2+/-4.2mmol/L vs. 12.8+/-4.7mmol/L, respectively) and 18 (9.6+/-2.9mmol/L vs. 10.4+/-4.0mmol/L, respectively) after AI. Mean PUN concentrations decreased significantly from Day 0 to Day 18 after AI in both pregnant and non-pregnant ewes. By using logistic regression analysis, there was no effect of PUN concentrations on the probability of pregnancy occurrence in the studied ewes (odds ratio: 0.97; 95% confidence interval: 0.9-1.05; P=0.45). In conclusion, there was no evidence of a relationship between PUN concentration and pregnancy rate for lactating AwassixMerino ewes in the present study because of low pregnancy rate observed. [less ▲]Detailed reference viewed: 63 (5 ULg)
Pregnancy-Associated Glycoprotein and Progesterone Concentrations during Pregnancy Failure in Bedouin Goat from the Southwest of Algeria.
; ; et al
in Reproduction in Domestic Animals (2010), 45(6), 213-238
Contents Thirteen female Bedouin goats living in arid land of Algeria Sahara desert were used in this study. These goats were pregnant but they sustained an abortion because of unidentified causes. None ... [more ▼]
Contents Thirteen female Bedouin goats living in arid land of Algeria Sahara desert were used in this study. These goats were pregnant but they sustained an abortion because of unidentified causes. None of the goats showed any signs of general disease. Plasma concentrations of caprine pregnancy-associated glycoproteins (cPAGs) and progesterone (P4) were determined during pregnancy using radioimmunoassay. The cPAGs concentration was undetectable (<0.8 ng/ml) throughout the first 2 weeks of gestation. From week 3 after mating, cPAGs concentration was detectable with significant individual variations (p < 0.05) reaching a maximum secretion (436.1 ng/ml). Throughout gestation, cPAGs concentration remained relatively constant but decreased few days before abortion, on an average of 9.2 +/- 1.2 days (n = 11), except for two females where the concentrations decreased later (1-2 days before abortion). One or two peaks of cPAGs concentrations (in 4/13 and in 9/13 females, respectively) have been measured few weeks before abortion (77-124 days after mating), when a decline of cPAGs was detected. The P4 concentration increased after mating, and was high from the first week till the end of pregnancy. The P4 concentration (9.1 +/- 0.9 ng/ml) decreased rapidly (<0.5 ng/ml) after 4 +/- 0.7 days (n = 6) or 9.4 +/- 1.6 days (n = 7) before abortion. A positive relationship (p < 0.01) was found between P4 and cPAGs concentrations during gestation. Results indicate that cPAGs and P4 measurements can be used for monitoring gestation and for abortion prediction. [less ▲]Detailed reference viewed: 140 (16 ULg)
Late embryonic and early fetal mortalities in dairy cows: the incidence and pregnancy protein profiles.
; ; et al
in Proceedings of the 20th International Congress of Hungarian Association for Buiatrics. (2010, October 20)Detailed reference viewed: 14 (2 ULg)
Effect of a corpus luteum with or without a cavity on fertility in dairy cows after prostaglandin treatment
; Melo de Sousa, Noelita ; Beckers, Jean-François et al
in Proceedings of the Hungarian Association for Buiatrics (2010, October)Detailed reference viewed: 11 (1 ULg)
Could Western blot analysis be an alternative to radio-immunoassay for sheep pregnancy associated glycoproteins measurements ?
; ; et al
in Reproduction in Domestic Animals (2010, September 15), 45(Suppl 3), 65
This study prospected whether Western blot analysis could be an alternative to radio-immunoassay (RIA) for sheep pregnancy diagnosis. Blood samples were taken from Boujaâd ewes (n= 9) and Boujaâd x D’man ... [more ▼]
This study prospected whether Western blot analysis could be an alternative to radio-immunoassay (RIA) for sheep pregnancy diagnosis. Blood samples were taken from Boujaâd ewes (n= 9) and Boujaâd x D’man (BD) cross breed (n= 12). The pregnancy associated glycoproeins (PAGs) were weekly monitored in plasma during the whole gestation and the first month postpartum by two RIA systems (El Amiri et al., 2007, Reprod Domest Anim 42:257-62). The highly immunoreactive samples derived from PAG profiles were tested using Western blot based on the same antiserum used in RIA. Each week (from the 13th to the 19th week), two pools of plasma samples (4 ml each) were taken from single and multiple lambing ewes respectively. Samples were submitted to a protein extraction at neutral pH, followed by ammonium sulfate (A.S.) precipitation, dialysis, and lyophilization. The placental extract was used as a check. Results revealed that ewes with multiple lambs presented numerically higher PAG concentrations. In both RIA systems, the high concentrations were recoded around the 19th week of gestation and the maximal concentrations varied from 120 to 700 ng/ml. In Western blot, the placental extracts reacted positively while the plasma samples did not give any positive reaction. In conclusion, the RIA remains the only sensitive method to measure PAGs in plasma. The present study strengthens the need to develop an ELISA kit as an alternative to RIA systems. [less ▲]Detailed reference viewed: 54 (2 ULg)
Monitoring endocrine profiles for prediction of stillbirth in Holstein-Friesian dairy cattle
; ; et al
Conference (2010, September 03)Detailed reference viewed: 12 (3 ULg)
Diagnostico precoz de gestation en Ganado ovino mediante un kit enzimoimmunoanalisis (EIA) de la glicoproteina asociada a la gestation (PAG) plasmatica.
; ; et al
in Proceedings of the XXXV Congreso de la Sociedad Espanola de Ovinotechnia y Caprinotecnia (SEOC) (2010, September)Detailed reference viewed: 20 (3 ULg)
Can we predict troubles during horse clinical examination by a simple test?
Peeters, Marie ; ; et al
Poster (2010, August 03)Detailed reference viewed: 45 (1 ULg)
Preliminary results obtained by ria determination of the proteins associated to pregnancy (PAG) in goat and sheep
; ; Beckers, Jean-François
in Annals of the Romanian Society for Cell Biology (2010), XV(1), 98-104
In goat and sheep, the glycoproteins associated to pregnancy (PAG) are little studied. Their study in these species is very important in the evaluation of prediction of early pregnancy. Even though ... [more ▼]
In goat and sheep, the glycoproteins associated to pregnancy (PAG) are little studied. Their study in these species is very important in the evaluation of prediction of early pregnancy. Even though numerous research has been done in order to investigate the physiological functions of placental proteins, the exact biofunction of the glycoproteins associated to pregnancy is still unknown. The objective of research was the study of the dynamics of PAG in goats and ewes in the first part of pregnancy(1-35 days after mating) and the determination of correlations with the reproductive status of females and to earlier the pregnancy.The experiment was realized on 49 Saanen x Carpatina goats and 72 Merinos of Palas sheep from the biostation of the Research and Development Institute for Goat and Sheep, Palas Constanta. The females in normal reproduction season were monitored for the detection of estrus and mated naturally. The blood was collected by puncture of the jugular vein on days 0, 7, 14, 25 and 35 after mating (day 0). The serum was obtained after centrifuging at 1500 x g for 15 minutes and stored at -20oC till the RIA determination. The RIA of the plasma with the purpose of detecting the concentration of proteins specific to pregnancy was realized in two experimental series in February and December 2008, at the Laboratory for Reproduction Physiology – The Faculty of Veterinary Medicine, Liege, Belgium. The RIA dosing was realized after the application of the method with the preincubation of the serums to test with specific serum (Atg°) and then with the marked antigen (Atg*), considering the high sensitivity of this method to detect the smallest values of PAG from day 0 to day 35 after mating. The diagnosis of the pregnancy state was based on the principle of PAG antibodies binding to the specific antigen, establishing through RIA the quantity of free antigen. Non-pregnancy involves the attachment of the antigen marked by non-specific antibodies. The preliminary results have demonstrated that the sheep PAG were had values ranging between 4.197-15.985 ng/ml and 0.01-3.39 ng/ml in nonpregnant ewes. In the pregnant goats, the PAG concentrations ranged between 16.75±3.44-27.17±2.95 ng/ml, while in the nonpregnant goats, the PAG values were 1.38±0.35 – 2.03±0.51 6 ng/ml. The purpose of the experiments was to find one accurate method for early pregnancy diagnosis in goat and sheep. The conclusion was that ovine pregnancy can be reliably diagnosed on Day 25 after AI and goat pregnancy on Day 30, by using a heterologous radioimmunoassay of PAG. [less ▲]Detailed reference viewed: 13 (0 ULg)
Efficiency of day seven collection of bovine embryosafter superovulation by flushing the oviducts and the uterin horns.
; ; Beckers, Jean-François et al
in Reproduction, Fertility and Development (2010), 22(1), 363Detailed reference viewed: 17 (7 ULg)
Effect of exogenous circulating anti-bPL antibodies on bovine placental lactogen measurements in foetal samples.
Alvarez Oxiley, Andrea Vivian ; Melo de Sousa, Noelita ; Hornick, Jean-Luc et al
in Acta Veterinaria Scandinavica (2010), 52
BACKGROUND: The involvement of placental lactogen (PL) in the regulation of foetal growth has been investigated in different species by in vivo immunomodulation techniques. However, when circulating ... [more ▼]
BACKGROUND: The involvement of placental lactogen (PL) in the regulation of foetal growth has been investigated in different species by in vivo immunomodulation techniques. However, when circulating antibodies are present together with the hormone, the procedure for hormonal measurement becomes considerably complex. The aim of this study was the immunoneutralization of bovine placental lactogen (bPL) concentrations in bovine foetal circulation by direct infusion of rabbit anti-bPL purified immunoglobulins (IgG) via a foetal catheter (in vivo study). The ability of a RIA based on guinea pig anti-bPL antiserum, for the measurement of bPL concentrations in samples containing exogenous rabbit anti-bPL immunoglobulins, was also analyzed in in vitro and in vivo conditions. METHODS: Six bovine foetuses were chronic cannulated on the aorta via the medial tarsal artery. Infusion of rabbit anti-bPL IgG was performed during late gestation. Pooled rabbit anti-bPL antisera had a maximal neutralization capacity of 25 microg bPL/mL of immunoglobulin. Interference of rabbit anti-bPL immunoglobulin with radioimmunoassay measurement using guinea pig anti-bPL as primary antibody was first evaluated in vitro. Polyclonal anti-bPL antibodies raised in rabbit were added in foetal sera to produce 100 samples with known antibodies titers (dilutions ranging from 1:2,500 till 1:1,280,000). RESULT(S): Assessment of the interference of rabbit anti-bPL antibody showed that bPL concentrations were significantly lower (P < 0.05) in samples added with dilutions of rabbit antiserum lower than 1:80,000 (one foetus) or 1:10,000 (four foetuses). It was also shown that the recovery of added bPL (12 ng/mL) was markedly reduced in those samples in which exogenous rabbit anti-bPL were added at dilutions lower than 1:20,000. Concentrations of foetal bPL were determined in samples from cannulated foetuses. In foetuses 1 and 6, bPL concentrations remained almost unchanged (<5 ng/mL) during the whole experimental period. In Foetus 3, bPL concentrations decreased immediately after IgG infusion and thereafter, they increased until parturition. CONCLUSION(S): The use of a bPL RIA using a guinea pig anti-bPL as primary antiserum allowed for the measurement of bPL concentrations in foetal plasma in presence of rabbit anti-bPL IgG into the foetal circulation. Long-term foetal catheterization allowed for the study of the influence of direct infusion of anti-bPL IgG on peripheral bPL concentrations in bovine foetuses. [less ▲]Detailed reference viewed: 114 (17 ULg)
Comparison of accuracy of sonography, rectal palpation, and pregnancy-associated glycoproteins tests for pregnancy diagnosis in cows
; ; Melo de Sousa, Noelita et al
in Proceedings of the XIth Middle European Buiatrics Congress (2010)Detailed reference viewed: 43 (2 ULg)
Prediction of stillbirth by monotoring endocrine and metabolic parameters in dairy cattle
; ; et al
in Reproduction in Domestic Animals (2010), 45(Suppl 3), 109Detailed reference viewed: 19 (2 ULg)
Le système de contrôle du chant des passériformes : un modèle d’étude de la plasticité neuronale
Boseret, Géraldine ; Beckers, Jean-François
in Annales de Médecine Vétérinaire (2010), 154(1), 48-60
In many songbirds species, wherein we can find the domestic canary (Serinus canaria), singing is generally produced either to defend a territory or to attract a mate. The Song Control System is a neural ... [more ▼]
In many songbirds species, wherein we can find the domestic canary (Serinus canaria), singing is generally produced either to defend a territory or to attract a mate. The Song Control System is a neural specialization, mostly located in telencephalic regions of the brain and associated to the control of song learning, perception and production. External factors such as testosterone (T), photoperiod and social cues have been described to modulate singing behaviour. Parallel to the song behaviour, some of the song control system nuclei (HVC, RA and Area X) demonstrate a puzzling amount of seasonal plasticity. Their volumes varies seasonally based on changes in cell spacing, neuropile size, dendritic arborisation and in the case on the nidopallial nucleus HVC on the incorporation of newborn neurons. We present here a review of this curious phenomenon of adult neuroplasticity associated to a complex behaviour, which doesn’t occur as far as we know in the mammalian adult brain [less ▲]Detailed reference viewed: 91 (9 ULg)
Monitoring endocrine profiles for prediction of stillbirth in Holstein-Friesian dairy cattle
; ; et al
Poster (2010)Detailed reference viewed: 29 (9 ULg)