Pregnancy diagnosis in cattle by using a new ELISA for pregnancy-associated glycoprotein: preliminary results.
Beckers, Jean-François ; Melo de Sousa, Noelita ; et al
in Proceedings of the 6th European Congress of Bovine Health and Management (ECBHM) (2011, September 07)
In the present study we report the use of a new PAG-ELISA for routine pregnancy diagnosis in cattle. A total of 2777 samples were collected from pregnant or non-pregnant cows and assayed by both classical ... [more ▼]
In the present study we report the use of a new PAG-ELISA for routine pregnancy diagnosis in cattle. A total of 2777 samples were collected from pregnant or non-pregnant cows and assayed by both classical PAG RIA-497 method and newly developed PAG-ELISA (Ref. code E.G.7, CER, Maloie). Pregnancy status was determined on the basis of RIA results. Samples were assumed to be issued from pregnant cows when PAG concentrations were higher than 0.8 ng/mL. PAG concentrations <0.8 ng/mL were considered negative (non-pregnant). PAG concentrations between 0.8 and 1.0 ng/mL were considered doubtful. With regard to ELISA, concentrations below 0.6 were considered negative. Concentrations between 0.6 and 0.8 ng/mL were considered doubtful and finally concentrations under 0.8 ng/mL were assumed as positive. Sensitivity(Se), specificity(Sp) and predictive values (PV) of pregnancy diagnosis were determined. A total of 143(5.2%), 137(4.9%) and 38(1.4%) samples gave doubtful results by RIA, ELISA or both systems, respectively. These samples (318/2777=11.5%) were not used for estimation of Se, Sp, PPV and NPV because clinical confirmation of pregnancy diagnosis could not be established by an additional method (rectal palpation, ultrasound or calving). Taking into account the remaining samples (2459), Se, Sp, PPV and NPV obtained by ELISA were respectively 98.9%, 89.9%, 98.7% and 96.4%. In conclusion, PAG-ELISA can be successfully used for pregnancy diagnosis in cattle. In the near future, further investigations are to be conducted in order to compare results obtained by ELISA test with those obtained by rectal palpation and ultrasound in field conditions. [less ▲]Detailed reference viewed: 53 (5 ULg)
Superovulation in the mare with commercially available pFSH
; Deleuze, Stefan ; Beckers, Jean-François et al
in Reproduction in Domestic Animals (2011), 46(s3), 138
Further studies on Stimufol with larger numbers, should be conducted as it might prove inexpensive and easy to superovulate mares.Detailed reference viewed: 95 (19 ULg)
Clinical implications of induced twin reduction in dairy cattle .
; ; et al
in Theriogenology (2011), 76(3), 512-521
Embryo reduction may prevent the negative effects of twinning in dairy cattle; however, the technique may carry an additional risk of pregnancy loss. The aim of this study was to evaluate the effect on ... [more ▼]
Embryo reduction may prevent the negative effects of twinning in dairy cattle; however, the technique may carry an additional risk of pregnancy loss. The aim of this study was to evaluate the effect on pregnancy maintenance of embryo reduction by manual amnion rupture in unilateral and bilateral twin pregnant cows. A secondary objective was to examine the dynamics of endocrine factors following the treatment. On Day 35-41 of gestation 55 cows bearing two live twin embryos (28 bilateral, 27 unilateral) were randomly assigned to a twin reduction group (n = 27; cows fitted with a progesterone releasing intra-vaginal device for 21 days after manual amnion rupture) or control group (n = 28; untreated cows). Pregnancy loss before Day 90 was recorded in nine control and eleven twin reduction cows (32.1% vs 40.7%, respectively, p = 0.508). Logistic regression models indicated that laterality was the only variable significantly affecting pregnancy loss. The pregnancy loss risk was 8.7 times higher for unilateral than for bilateral twin pregnancies (59.3% vs 14.3%, respectively, P < 0.001) yet was similar in the unilateral control and unilateral twin reduction cows (62.3% vs 53.8%, respectively, P = 0.581). In contrast, four of 14 cows with bilateral twin pregnancies undergoing twin reduction lost their pregnancies while no losses were recorded in control cows with bilateral pregnancies (P = 0.049). A rise in plasma progesterone concentration was detected on the day following treatment in the twin reduction group and concentrations remained high within the first week of treatment. Plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations fell between Day 35-41 and Day 42-48, regardless of treatment. Our findings indicate that embryo reduction by manual amnion rupture did not carry an additional risk of pregnancy loss for unilateral twin pregnancies, whereas it increased the risk of pregnancy failure in bilateral twin pregnancies. However, benefits of preventing cows from delivering twins might also be considered when assessing the success of embryo reduction in bilateral twin pregnancies. [less ▲]Detailed reference viewed: 21 (2 ULg)
In vivo oocyte developmental competence is reduced in lean but not in obese superovulated dairy cows after intraovarian administration of IGF1.
; ; et al
in Reproduction (Cambridge, England) (2011), 142(1), 41-52
The present study investigated the role of IGF1 in lactating lean and non-lactating obese dairy cows by injecting 1 μg IGF1 into the ovaries prior to superovulation. This amount of IGF1 has been linked ... [more ▼]
The present study investigated the role of IGF1 in lactating lean and non-lactating obese dairy cows by injecting 1 μg IGF1 into the ovaries prior to superovulation. This amount of IGF1 has been linked with pregnancy loss in women with the polycystic ovary syndrome (PCOS) and was associated with impaired bovine oocyte competence in vitro. Transcript abundance and protein expression of selected genes involved in apoptosis, glucose metabolism, and the IGF system were analyzed. Plasma concentrations of IGF1 and leptin, and IGF1 in uterine luminal fluid (ULF), were also measured. IGF1 treatment decreased embryo viability in lean cows to the levels observed in obese cows. Obese cows were not affected by IGF1 treatment and showed elevated levels of IGF1 (in both plasma and ULF) and leptin. Blastocysts from lean cows treated with IGF1 showed a higher abundance of SLC2A1 and IGFBP3 transcripts. IGF1 treatment reduced protein expression of tumor protein 53 in blastocysts of lean cows, whereas the opposite was observed in obese cows. IGF1 in plasma and ULF was correlated only in the control groups. Blastocyst transcript abundance of IGF1 receptor and IGFBP3 correlated positively with IGF1 concentrations in both plasma and ULF in lean cows. The detrimental microenvironment created by IGF1 injection in lean cows and the lack of effect in obese cows resemble to a certain extent the situation observed in PCOS patients, where IGF1 bioavailability is increased in normal-weight women but reduced in obese women, suggesting that this bovine model could be useful for studying IGF1 involvement in PCOS. [less ▲]Detailed reference viewed: 10 (2 ULg)
Effect of a combined iodine and selenium supplementation on I and Se status of cows and their calves
Guyot, Hugues ; ; Ramery, Eve et al
in Journal of Trace Elements in Medicine and Biology (2011), 25(2), 118-124
Iodine (I) and selenium (Se) deficiencies are commonly reported in cattle, however, there are also studies regarding a very high iodine supply. The aim of the study was to determine the long-term effect ... [more ▼]
Iodine (I) and selenium (Se) deficiencies are commonly reported in cattle, however, there are also studies regarding a very high iodine supply. The aim of the study was to determine the long-term effect of I and Se supplementation on non-pregnant cows, pregnant cows and their calves. The hypothalamus pituitary axis was investigated (TSH, T4, T3 assays) during a TRH challenge on non-pregnant cows. Twenty-four cows, half of them pregnant, were assigned into 2 diet-groups, one group with a low I (0.45 ppm) and Se (0.15 ppm) diet (LISe), the other with a high I (5.45 ppm) and Se (0.45 ppm) diet (HISe), for a period of 120 days. Nutritional (plasma iodide, urinary I, plasma Se, I content in colostrum and foetal fluids) and functional (thyrotropin, thyroid hormones, glutathione-peroxidase activity in erythrocytes) markers of I and Se status were assayed in dams at regular intervals for 120 days and in their calves at birth. A TRH challenge was performed on 8 non-pregnant cows at day 110 of the trial. At the end of the study, I and Se nutritional markers were higher in dams in the HISe group, compared to the LISe group, except for plasma Se. At birth, I nutritional markers in calves in the HISe group were higher compared to the LISe group. Reactivity of the pituitary–thyroid-axis was not influenced by I and Se supplementation. I and Se supplementation is efficient in improving newborn status. [less ▲]Detailed reference viewed: 70 (17 ULg)
Contrôle de l'involution utérine au moyen de l'écographie transabdominale en mode B : validation de la technique
; ; Farnir, Frédéric et al
in Colloques de la 43e Journée de la Recherche Porcine (2011, February 15)Detailed reference viewed: 20 (2 ULg)
Comparison between blood serum and salivary cortisol concentrations in horses using an adrenocorticotropic hormone challenge
Peeters, Marie ; ; Beckers, Jean-François et al
in Equine Veterinary Journal (2011), 43(4), 487-493Detailed reference viewed: 25 (2 ULg)
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: 14 (5 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: 38 (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: 17 (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: 51 (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: 10 (0 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: 35 (4 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: 123 (15 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: 5 (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: 40 (2 ULg)
Monitoring endocrine profiles for prediction of stillbirth in Holstein-Friesian dairy cattle
; ; et al
Conference (2010, September 03)Detailed reference viewed: 10 (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: 8 (3 ULg)