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See detailHigh concentrations of Myeloperoxidase in the equine uterus as an indicator of endometritis
Parrilla Hernandez, Sonia ULg; Ponthier, Jérôme ULg; Franck, Thierry ULg et al

in Theriogenology (in press)

Intra-luminal fluid and excessive abnormal hyper-edema are regularly used for the diagnosis of endometritis in the mare, which is routinely confirmed by the presence of neutrophils on endometrial smears ... [more ▼]

Intra-luminal fluid and excessive abnormal hyper-edema are regularly used for the diagnosis of endometritis in the mare, which is routinely confirmed by the presence of neutrophils on endometrial smears. Studies show a relation between neutrophils and myeloperoxidase (MPO), an enzyme contained in and released by neutrophils during degranulation or after cell lysis. This enzyme has been found in many fluids and tissues and associated with different inflammatory pathologies in the horse. The aims of this study were to assess the presence and concentration of MPO in the equine uterus, and to investigate its relation with neutrophils, and other clinical signs of endometritis. Mares (n=51) were evaluated for presence of intra-luminal fluid and excessive endometrial edema before breeding, and a small volume lavage and cytology samples were obtained. From 69 cycles, supernatant of the uterine flushes was analysed with a specific equine MPO ELISA assay to measure MPO concentration. Cytology samples were used for the diagnosis of endometritis. MPO was present in the uterus of all estrus mares in highly variable concentrations. MPO concentrations were significantly (p<0.05) higher in samples with positive cytologies and in presence of intra-luminal fluid. Occasionally, some samples with negative cytologies showed high MPO concentration, but the opposite was never observed. Cycles presenting hyper-edema weren’t associated to high concentration of MPO, intra-luminal fluid or positive cytology making it a poor diagnostic tool of endometritis. [less ▲]

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See detailIn vitro embryo production in goats: slaughterhouse and laparoscopic ovum pick up 2 (LOPU) derived oocytes have different kinetics and requirements regarding maturation 3 media
Souza-Fabjan, JM; Locatelli, Y; Duffard, N et al

in Theriogenology (2014), 81

A total of 3427 goat oocytes were used in this study in order to identify possible 24 differences during in vitro embryo production from slaughterhouse or laparoscopic ovum 25 pick up (LOPU) oocytes. In ... [more ▼]

A total of 3427 goat oocytes were used in this study in order to identify possible 24 differences during in vitro embryo production from slaughterhouse or laparoscopic ovum 25 pick up (LOPU) oocytes. In Experiment 1, one complex, one semi defined and one simplified 26 in vitro maturation (IVM) media were compared using slaughterhouse oocytes. In 27 Experiment 2, we checked the effect of oocyte origin (slaughterhouse or LOPU) on the 28 kinetics of maturation (18 vs. 22 vs. 26 h) when submitted to semi defined or simplified 29 media. In Experiment 3, we determined the differences in embryo development between 30 slaughterhouse and LOPU oocytes when submitted to both media and then to in vitro 31 fertilization (IVF) or parthenogenetic activation (PA). Embryos from all groups were vitrified 32 and their viability evaluated in vitro after thawing. In Experiment 1, no difference (P>0.05) 33 was detected among treatments for maturation rate (MII; 88% on average), cleavage (72%), 34 blastocyst from the initial number of cumulus oocyte complexes (COC; 46%) or from the 35 cleaved ones (63%), hatching rate (69%) and the total number of blastomeres (187). In 36 Experiment 2, there was no difference of MII rate between slaughterhouse oocytes cultured 37 for 18 or 22 h, whereas the MII rate increased significantly (P<0.05) between 18 and 22 h for 38 LOPU oocytes in the simplified medium. Moreover, slaughterhouse oocytes cultured in 39 simplified medium matured significantly faster than LOPU oocytes at 18 and 22 h (P<0.05). 40 In Experiment 3, cleavage rate was significantly greater (P<0.001) in all four groups of 41 embryos produced by PA than IVF. Interestingly, PA reached similar rates for slaughterhouse 42 oocytes cultured in both media, but improved (P<0.05) cleavage rate of LOPU oocytes. 43 Slaughterhouse oocytes had acceptable cleavage rate after IVF (~67%), whereas LOPU 44 oocytes displayed a lower one (~38%), in contrast to cleavage after PA. The percentage of 45 blastocysts in relation to cleaved embryos was not affected by the origin of the oocytes 46 (P>0.05). Therefore, slaughterhouse oocytes developed a greater proportion of blastocysts than LOPU ones, expressed as the percentage of total COC entering to IVM. Vitrified-thawed 48 blastocysts presented similar survival and hatching rates between the oocyte origin, media or 49 method of activation. In conclusion, slaughterhouse and LOPU derived oocytes may have 50 different IVM kinetics and require different IVM-IVF conditions. Although the IVM and IVF 51 systems still need improvements in order to enhance embryo yield, the in vitro development 52 (IVD) step is able to generate good quality embryos from LOPU derived oocytes. [less ▲]

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See detailEffect of non-sperm cells removal with single layer colloidal centrifugation on myeloperoxidase concentration in post-thaw equine semen
Ponthier, Jérôme ULg; Teague, Sheila; Franck, Thierry ULg et al

in Theriogenology (2013), 80(9), 1082-1087

Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in, and released by, neutrophils during degranulation or after lysis. Post-thaw semen contains MPO and its concentration is associated with ... [more ▼]

Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in, and released by, neutrophils during degranulation or after lysis. Post-thaw semen contains MPO and its concentration is associated with decreased sperm motility. Recently, MPO concentration in post-thaw semen was shown to be associated with presence of non-sperm cells. The aim of this study was to describe the effect of centrifugal fractionation of semen prior to cryopreservation on post-thaw concentrations of non-sperm cells and MPO. The experimental design consisted in freezing semen with or without previous centrifugation through two concentrations of single layer colloid media. Non-sperm cells and MPO concentrations were assessed in pellet and upper layer at each step of the procedure and MPO was detected in cells by immunocytochemistry. Single layer colloid centrifugation decreased non-sperm cells and MPO concentrations in post-thaw semen. The MPO concentration was correlated with concentration of non-sperm cells in the upper layer of the supernatant. In post-thaw semen, with or without previous single layer colloid centrifugation, MPO concentration was correlated with concentration of non-sperm cells. Overall, neutrophils were rarely observed and non-sperm cells were mainly epithelial cells or cellular debris, as demonstrated by MPO immunocytochemistry. Following single layer colloid centrifugation, MPO concentration was decreased, and correlations observed in different samples of the experiments revealed an association between concentrations of MPO and non-sperm cells. At all steps of the semen processing and cryopreservation, MPO immunostaining was clearly identified only on non-sperm cells. Our results demonstrate that non-sperm cells present in fresh semen release MPO during freezing. [less ▲]

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See detailIn vitro oocyte IGF-I priming increases inner cell mass proliferation of in vitro-formed bovine blastocysts
Velazquez, MA; Hadeler, KG; Herrmann, D et al

in Theriogenology (2012), 78(3), 517-527

Studies addressing the effects of supraphysiological levels of IGF-1 on oocyte developmental competence are relevant for unravelling conditions resulting in high bioavailability of IGF-1, such as the ... [more ▼]

Studies addressing the effects of supraphysiological levels of IGF-1 on oocyte developmental competence are relevant for unravelling conditions resulting in high bioavailability of IGF-1, such as the polycystic ovary syndrome (PCOS). This study investigated the effects of supraphysiological levels of IGF-1 during in vivo folliculogenesis on the morula-blastocyst transition in bovine embryos. Compacted morulae were non-surgically collected and frozen for subsequent mRNA expression analysis (IGF1R, IGBP3, TP53, AKT1, SLC2A1, SLC2A3, and SLC2A8), or underwent confocalmicroscopy analysis for protein localization (IGF1R and TP53), or were cultured in vitro for 24 h. In vitro-formed blastocysts were subjected to differential cell staining. The mRNA expression of SLC2A8 was higher in morulae collected from cows treated with IGF-1. Both IGF1R and TP53 protein were present in the plasma membrane and cytoplasm. IGF-1 treatment did not affect protein localization of both IGF1R and TP53. In vitro-formed blastocysts derived from morulae recovered from IGF-1-treated cows displayed a higher number of cells in the inner cell mass (ICM). Total cell number (TCN) of in vitro-formed blastocysts was not affected. A higher mean ICM/TCN proportion was observed in in vitro-formed blastocysts derived from morulae collected from cows treated with IGF-1. The percentage of in vitro-formed blastocysts displaying a low ICM/TCN proportion was decreased by IGF-1 treatment. In vitro-formed blastocysts with a high ICM/TCN proportion were only detected in IGF-1 treated cows. Results show that even a short in vivo exposure of oocytes to a supraphysiological IGF-1 microenvironment can increase ICM cell proliferation in vitro during the morula to blastocyst transition [less ▲]

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See detailThe inseminating bull and plasma pregnancy-associated glycoprotein levels were related to peripheral leukocyte counts during the late pregnancy/early postpartum period in high-producing dairy cows.
Abdelfatah-Hassan, A.; Almeria, S.; Serrano, B. et al

in Theriogenology (2012), 77(7), 1390-1397

The interest in the immunology and endocrinology aspects of the peripartum in dairy cows has been increasing in the last years, since the animal status during the pre- and postpartum period determines the ... [more ▼]

The interest in the immunology and endocrinology aspects of the peripartum in dairy cows has been increasing in the last years, since the animal status during the pre- and postpartum period determines the subsequent productive and reproductive performance of the animal. At parturition, a drop in the immune functions of the peripheral blood polymorph-nuclear cells (PMN) was observed after a peak of Pregnancy-Associated Glycoproteins (PAGs) and recently, the inseminating bull was found to affect the plasma levels of bovine Pregnancy-Associated Glycoproteins (bovine PAGs) in the pregnant Holstein-Friesian dairy cows. The present work was aimed to determine whether changes in leukocyte counts during the peripartum, indicative of the animal immune status, could be related to the inseminating bull and to the levels of plasma pregnancy-associated glycoproteins (PAGs). Ninety six clinically healthy, single pregnant cows were selected in a commercial dairy herd. Six blood samples (four prepartum and two postpartum) were obtained every two weeks, and total and differential blood cell counts were analyzed. Based on procedures of General Linear Model analysis of variance for repeated measures, the inseminating bull affected counts of total leukocytes and lymphocytes (P<0.001; between-subject effects) throughout the peripartum period. In addition, cows with high plasma levels of PAGs (>900 ρg/µL) on Day 262-268 of gestation had higher numbers of total leukocytes and neutrophils throughout the peripartum period (P<0.001; between-subject effects). Young animals (≤ 1 lactation) had higher total leukocyte and lymphocytes counts than older cows (two or more lactations) throughout the study period. These results showed a clear relationship between the inseminating bull and plasma PAG levels and peripheral leukocyte counts during the peripartum period of dairy cows. [less ▲]

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See detailSperm quality analysis in XX, XY and YY males of the Nile tilapia (Oreochromis niloticus).
Gennotte, Vincent ULg; François, Ekniel; Rougeot, Carole ULg et al

in Theriogenology (2012)

In Nile tilapia (Oreochromis niloticus), individuals with atypical sexual genotype are commonly used in farming (use of YY males to produce all-male offsprings), but they also constitute major tools to ... [more ▼]

In Nile tilapia (Oreochromis niloticus), individuals with atypical sexual genotype are commonly used in farming (use of YY males to produce all-male offsprings), but they also constitute major tools to study sex determinism mechanisms. In other species, sexual genotype and sex reversal procedures affect different aspects of biology such as growth, behaviour and reproductive success. The aim of this study was to assess the influence of sexual genotype on sperm quality in Nile tilapia. Milt characteristics were compared in XX (sex-reversed), XY and YY males in terms of gonadosomatic index, sperm count, sperm motility and duration of sperm motility. Sperm motility was measured by computer-assisted sperm analysis (CASA) quantifying several parameters: total motility, progressive motility, curvilinear velocity, straight line velocity, average path velocity and linearity. None of the sperm trait measured differed significantly between the three genotypes. Mean values of gonadosomatic index, sperm concentration and sperm motility duration of XX, XY and YY males respectively ranged from 0.92 to 1.33 %, from 1.69 to 2.22 × 10(9) cells mL-1 and from 18’04’’ to 27’32’’. Mean values of total motility and curvilinear velocity 1 min after sperm activation respectively ranged from 53 to 58 % and from 71 to 76 µm s-1 for the three genotypes. After 3 min of activity, all the sperm motility and velocity parameters dropped by half and continued to slowly decrease thereafter. Seven min after activation, only 9 to 13 % of spermatozoa were still progressive. Our results prove that neither sexual genotype nor hormonal sex reversal treatments affect sperm quality in male Nile tilapias with atypical sexual genotype. [less ▲]

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See detailEffect of restraint stess on plasma concentrations of cortisol, progesterone and pregnancy associated-glycoproteins-1 in pregnant heifers during late embryonic development.
Szenci, O.; Karen, A.; Bajcsy, A. C. et al

in Theriogenology (2011), 76(8), 1380-1385

The aim of the present study was to evaluate the effect of restraint stress, which is commonly practised in the field, on plasma concentrations of cortisol, progesterone (P4) and bovine pregnancy ... [more ▼]

The aim of the present study was to evaluate the effect of restraint stress, which is commonly practised in the field, on plasma concentrations of cortisol, progesterone (P4) and bovine pregnancy-associated glycoprotein-1 (boPAG-1) in pregnant heifers between Days 30 to 40 of gestation. Twelve Holstein-Friesian heifers between Days 30 (Day 0 of experiment) and 40 (Day 10 of experiment) of pregnancy in a Hungarian dairy farm were used in the present study. The heifers were exposed to an acute stressor consisting of immobilisation (restraint stress) in a crush for 2 h (Group 1, n = 6) on Day 2 (Hour 48) and for 2 × 2 h (Group 2, n = 6) on Days 2 and 3 (Hour 72) of the experiment. Transrectal ultrasonography (7.5 MHz linear-array rectal transducer) was performed daily from Day 0 to Day 10 of the experiment to detect embryonic heartbeat or the fate of the conceptus. Blood samples were withdrawn before each ultrasonographic examination. Additional blood samples were withdrawn by 1 and 2 h (at Hours 49 and 50 in Groups 1 and 2 and Hours 73 and 74 in Group 2) of the onset of applying the stressor. Plasma cortisol, P4 and boPAG-1 concentrations were measured by radioimmunoassay. Acute restraint stress significantly (P < 0.001) increased the plasma cortisol level in pregnant heifers at 1 h of the exposure to the stressor at Days 2 (48 h) and 3 (72 h) of the experiment. On the other hand, the restraint stress did not affect the concentration of P4 and boPAG-1 concentrations in both groups. In conclusion, restraint stress for 2 h during early pregnancy in heifers increased blood cortisol, but it did not affect the concentrations of P4 and boPAG-1 between Days 30 to 40 of gestation. [less ▲]

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See detailClinical implications of induced twin reduction in dairy cattle
Andreu-Vazquez, C.; Garcia-Ispierto, I.; Lopez-Bejar, M. 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 ▲]

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See detailAltered secretion of pregnancy-associated glycoproteins during gestation in bovine somatic clones.
Constant, Fabienne; Camous, Sylvaine; Chavatte-Palmer, Pascale 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 ▲]

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See detailNeospora caninum and coxiella burnetii seropositivity are related to endocrine pattern changes during gestation in lactating dairy cows.
Garcia-Ispierto, I.; Nogareda, C.; Yaniz, J. L. et al

in Theriogenology (2010), 74(2), 212-230

Q fever is a zoonotic infection caused by Coxiella burnetii that is endemic worldwide. Domestic ruminants are a source of infection for humans. Given the suggestion that the bacterium recrudesces during ... [more ▼]

Q fever is a zoonotic infection caused by Coxiella burnetii that is endemic worldwide. Domestic ruminants are a source of infection for humans. Given the suggestion that the bacterium recrudesces during pregnancy in cattle, this study was designed to determine whether C. burnetii infection affects hormonal patterns, such as progesterone, cortisol, pregnancy associated glycoproteins (PAG), and prolactin during gestation in lactating cows. Possible interactions with Neospora caninum were also explored. The study was performed on 58 gestating non-aborting cows. Blood samples for hormone determinations were collected on Days 40, 90, 120, 150, 180, and 210 of gestation. For antibody determinations, blood was collected at day 40 postinsemination and postpartum. By GLM repeated measures analysis of variance, we established the effects of production and reproductive variables as well as Coxiella and Neospora seropositivity related to changes on cortisol, PAG, progesterone, and prolactin levels. Coxiella antibody levels were significantly related to cortisol, PAG, and plasma progesterone concentrations, whereas Neospora seropositivity was linked to plasma progesterone concentrations. The interaction between Coxiella and Neospora seropositivity was correlated with cortisol and plasma progesterone levels, whereas the interaction seropositivity against C. burnetii-plasma cortisol concentration was related to plasma PAG levels. Finally, an effect of lactation number only was observed on plasma prolactin. Our findings suggest that both the N. caninum and C. burnetii infection or the presence of both modify endocrine patterns throughout gestation. Cows seropositive to both, Neospora and Coxiella, showed higher plasma progesterone levels than the remaining animals examined. Seropositivity to C. burnetii was associated with placental damage and diminishing PAG levels throughout the second half of gestation, along with increased plasma cortisol levels on Day 180 of gestation. [less ▲]

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See detailWhen is a cow in estrus? Clinical and practical aspects.
Roelofs, Judith; Lopez-Gatius, Fernando; Hunter, R. H. F. et al

in Theriogenology (2010)

Good detection of estrus is critically important in dairy husbandry. Incorrect detection of estrus is related to loss of profit due to extended calving intervals, milk loss, veterinary costs, etc ... [more ▼]

Good detection of estrus is critically important in dairy husbandry. Incorrect detection of estrus is related to loss of profit due to extended calving intervals, milk loss, veterinary costs, etc. Detection of estrus remains a major problem despites enormous progress in the knowledge of reproductive physiology of the cow and in development of estrus detection aids. To achieve good estrus detection, many factors have to be taken into account. On one hand a cow has to express estrus and on the other hand the farmer has to detect it. Combined action of several hormones causes physiological changes that lead to ovulation and an environment in the uterus that allows sperm to fertilize the egg. Besides these internal actions, a number of external changes can be observed. When using visual observations, time of the day and time spend on observation have a great impact on detection rates. Many devices are available to aid in estrus detection, such as pedometers, mount devices, temperature, and hormone measurements. Expression of estrus can be influenced by many factors. Heritability, number of days postpartum, lactation number, milk production, and health are known to influence estrus expression. Environmental factors like nutrition, season, housing, herd size, etc. also play a role in estrus expression. To evaluate estrus detection, record keeping is very important; a number of formulas can be used to assess detection efficiency. Besides the farmer, the veterinarian and inseminator can play an important role in estrus confirmation and good insemination strategy. In the end, the time of ovulation and the age of the egg at sperm penetration is critical for conception. Therefore, emphasis in research needs to be on the timing of insemination relative to ovulation, and thus on the detection of ovulation [less ▲]

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See detailSubclinical infection and periodic shedding of equid herpesvirus 3
Barrandeguy, Maria; Vissani, A.; Pont Lezica, F. et al

in Theriogenology (2010), 74

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See detailEfficiency of embryonic development after Intra-Follicular and Intra-Oviductal transfer of in vitro and in vivo matured horse oocytes
Deleuze, Stefan ULg; Goudet, Ghylène; Caillaud, Maud et al

in Theriogenology (2009), 72(2), 203

In vivo techniques, such as intra-oviductal Oocyte Transfer (OT) and Intra-Follicular Oocyte Transfer (IFOT) can be considered as alternatives to bypass the lack of efficient superovulation treatments and ... [more ▼]

In vivo techniques, such as intra-oviductal Oocyte Transfer (OT) and Intra-Follicular Oocyte Transfer (IFOT) can be considered as alternatives to bypass the lack of efficient superovulation treatments and the inadequacy of conventional in vitro fertilization techniques in the horse. We compared embryo production following transfer of in vivo recovered oocytes (1) into a recipient’s oviduct or (2) into her preovulatory follicle either immediately after ovum pick up or (3) after in vitro maturation. Recipients were inseminated with fresh semen of a stallion with a known normal fertility. Ten days after surgery, rates of embryos collected in excess to the number of ovulations were calculated and compared for each group. Embryo collection rates were 32.5% (13/40), 5.5% (3/55) and 12.8% (6/47) for OT, post-IVM and immediate IFOT respectively. OT significantly yielded more embryos than immediate and post-IVM IFOT did. We also showed that in vitro matured oocytes could succesfully be used for IFOT. Our results also suggest that improvement of the IFOT technique could turn it into an inexpansive and easy to perform procedure that could be an answer to the inefficiency of superovulation treatments in the mare. [less ▲]

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See detailFirst birth of an animal from an extinct subspecies (Capra pyrenaica pyrenaica) by cloning
Folch, J.; Cocero, M. J.; Chesne, P. et al

in Theriogenology (2009), 71

Two experiments have been performed to clone the bucardo, an extinct wild goat. The karyoplasts were thawed fibroblasts derived from skin biopsies, obtained and cryopreserved in 1999 from the last living ... [more ▼]

Two experiments have been performed to clone the bucardo, an extinct wild goat. The karyoplasts were thawed fibroblasts derived from skin biopsies, obtained and cryopreserved in 1999 from the last living specimen, a female, which died in 2000. Cytoplasts were mature oocytes collected from the oviducts of superovulated domestic goats. Oocytes were enucleated and coupled to bucardo's fibroblasts by electrofusion. Reconstructed embryos were cultured for 36h or 7d and transferred to either Spanish ibex or hybrid (Spanish ibex malex domestic goat) synchronized recipients. Embryos were placed, according to their developmental stage, into the oviduct or into the uterine horn ipsilateral to an ovulated ovary. Pregnancy was monitored through their plasmatic PAG levels. In Experiment 1, 285 embryos were reconstructed and 30 of them were transferred at the 3- to 6-cells stage to 5 recipients. The remaining embryos were further cultured to day 7, and 24 of them transferred at compact morula/blastocyst stage to 8 recipients. In Experiment 2, 154 reconstructed embryos were transferred to 44 recipients at the 3- to 6-cells stage. Pregnancies were attained in 0/8 and 7/49 of the uterine and oviduct-transferred recipients, respectively. One recipient maintained pregnancy to term, displaying very high PAG levels. One morphologically normal bucardo female was obtained by caesarean section. The newborn died some minutes after birth due to physical defects in lungs. Nuclear DNA confirmed that the clone was genetically identical to the bucardo's donor cells. To our knowledge, this is the first animal born from an extinct subspecies. [less ▲]

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See detailPregnancy patterns during the early fetal period in high producing dairy cows treated with GnRH or progesterone.
Bech-Sabat, G.; Lopez-Gatius, F.; Garcia-Ispierto, I. et al

in Theriogenology (2009), 71

In order to explore pregnancy patterns in high producing dairy cows treated with GnRH or progesterone at pregnancy diagnosis (Days 28-34), two consecutive experiments were designed. In Experiment 1, cows ... [more ▼]

In order to explore pregnancy patterns in high producing dairy cows treated with GnRH or progesterone at pregnancy diagnosis (Days 28-34), two consecutive experiments were designed. In Experiment 1, cows bearing a single embryo were randomly assigned to a PRID (n=40; cows fitted with a progesterone releasing intra-vaginal device for 28 days), GnRH (n=40; cows receiving GnRH) or Control (n=26; untreated cows) group. PRID treatment led to a rise in plasma progesterone concentrations in the 7 days following the onset of treatment compared to the other two groups. In Experiment 2, in which we also examined twin pregnancies, animals were randomly assigned to PRID (n=312) or GnRH (n=294) treatment groups. Treatments were the same as described for Experiment 1. Logistic regression procedures revealed that in cows with a single corpus luteum, the probability of pregnancy loss between the first (Days 28-34) and second (Days 65-62) pregnancy diagnosis decreased by a factor of 0.51 in the PRID group compared to the GnRH group. However, in cows with two or more corpora lutea, PRID treatment increased the likelihood of pregnancy loss by a factor of three, compared to GnRH treatment. In cows carrying twins, the conceptus reduction rate was higher (P=0.02) for the GnRH (36%) than for the PRID (16.4%) group. Formation of a new corpus luteum was recorded in 17.7% of cows in the GnRH group. Our results indicate that compared to GnRH treatment, progesterone treatment given at pregnancy diagnosis in high producing dairy cows, reduced by a factor of 0.51 and increased by a factor of 3 the probability of pregnancy loss in cows with a single or with two or more corpora lutea, respectively, and reduced the conceptus reduction rate in cows carrying twins. The practical implications of our findings are that in herds with a high incidence of early fetal loss of a non-infectious nature, treatment at the time of pregnancy diagnosis with PRID in cows with one corpus luteum and with GnRH in cows with two or more corpora lutea should offer considerable benefits. [less ▲]

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See detailComparison of accuracy of ultrasonography, progesterone and pregnancy-associated 4 glycoprotein tests for pregnancy diagnosis in semi-domesticated reindeer
Savela, Hannele; Vahtiala, S.; Lindeberg, H. et al

in Theriogenology (2009), 72

The aim of the study was to compare transrectal ultrasound with progesterone (P4) and pregnancy-associated glycoproteins (PAG) as pregnancy detection methods for semi-domesticated reindeer in field ... [more ▼]

The aim of the study was to compare transrectal ultrasound with progesterone (P4) and pregnancy-associated glycoproteins (PAG) as pregnancy detection methods for semi-domesticated reindeer in field conditions. Female reindeer (n=195) were scanned transrectally by a 7.5 MHz linear array transducer and blood sampled either in Dec 2005 (n=33), Dec 2006 (n=92) or Jan 2007 (n=70) during early- or mid-gestation. Plasma levels of P4 and PAG were assessed by RIA. Based on calving records, the sensitivity, specificity, predictive values and the overall accuracy of the three tests were calculated. The overall calving rate calculated from the calving records was 86.2%. The overall accuracy of transrectal ultrasound was 99.5%. The sensitivity and specificity of transrectal ultrasound were 99.4% and 100%, respectively. In the plasma P4 test, the threshold level of 5.5 nmol/L gave the highest overall accuracy (93.3%). The sensitivity of the P4 test decreased from 96.4% to 81.5%, when the threshold level increased from 5.0 nmol/L to 8.0 nmol/L, while the specificity remained at 85.2% over the range of these cut-off values. The overall accuracy of the plasma PAG test decreased from 96.4% to 64.1% when the plasma PAG threshold level increased from 0.5 ng/mL to 3.5 ng/mL, whereas sensitivity decreased from 99.4% to 58.3%. Specificity increased from 77.8% to 100% when the plasma PAG threshold level reached 3.0 ng/mL. Transrectal ultrasound showed higher diagnostic values than plasma P4-RIA and PAG-RIA in diagnosing pregnancy of reindeer, with the advantage that diagnoses can be made in real-time in field conditions [less ▲]

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See detailValidation of a new pregnancy-associated glycoprotein radioimmunoassay method for the detection of early pregnancy in ewes.
Barbato, Olimpia; Melo de Sousa, Noelita ULg; Debenedetti, A. et al

in Theriogenology (2009), 72

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 ... [more ▼]

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 [less ▲]

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See detailDetection of viral genomes of caprine arthritis-encephalitis virus (CAEV) in semen and in genital tract tissues of male goat
Ali Al Ahmad, M. Z.; Fieni, F.; Pellerin, J. L. et al

in Theriogenology (2008)

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See detailLack of risk of transmission of caprine arthritis-encephalitis virus (CAEV) after an appropriate embryo transfer procedure
Ali Al Ahmad, M. Z.; Chebloune, Y.; Bouzar, Amel ULg et al

in Theriogenology (2008)

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See detailSet up of a serum-free culture system for bovine embryos: embryo development and quality before and after transient transfer
George, F.; Daniaux, C.; Génicot, G. et al

in Theriogenology (2008), 69

It is well known that serum in culture medium negatively affects blastocyst quality. The objective of this work was to develop and test a serum-free culture medium which could improve embryo quality ... [more ▼]

It is well known that serum in culture medium negatively affects blastocyst quality. The objective of this work was to develop and test a serum-free culture medium which could improve embryo quality, measured by the resistance to freezing, lipid and glutathione content of the resulting blastocysts, as well as the ability of the blastocysts to elongate after transient transfer to recipient cows. In a first experiment we showed that adding a mixture of insulin, transferrin and selenium to serum-free Synthetic Oviduct Fluid medium (SOF–ITS) improved embryo development and quality. In the second experiment, the addition of BSA to SOF–ITS further improved blastocyst development. Moreover, a reduction in lipid content of morulae was observed in SOF–ITS–BSA by comparison with morulae cultured with serum (SOF–FCS). The resistance to freezing measured by hatching rates 24 h postthawing was also improved for blastocysts with a diameter between 160 and 180 mm cultured in SOF–ITS–BSA by comparison to those produced with serum. In order to evaluate the redox potential of the embryos, reduced glutathione content (GSH) was evaluated both before and after cryopreservation. A significant decrease in glutathione was observed after freezing, whatever the culture medium, but no difference was observed between culture conditions. Transient transfers were performed and elongated D- 13 embryos were recovered. Elongation was more pronounced and the embryonic disk more often visible in embryos cultured in SOF–ITS–BSA than in embryos cultured with FCS. In conclusion, the serum-free system we developed to produce in vitro bovine embryos meets the developmental and qualitative requirements for a large-scale use. [less ▲]

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