Fertility prediction in dairy goats from Murciano-Granadina breed: The role of sperm evaluation and female traits.

Fertility is one of the most economically important traits in farm animals, due to the direct and indirect costs associated to low pregnancy rates. Thus, one of the priority goals in animal reproduction is to predict the performance that the semen doses will have in vivo based on the quality values obtained in laboratory assays. Attempts have been made for getting a predictive model of fertility of frozen-thawed sperm in dairy goats, but similar studies have not been conducted for chilled goat buck sperm doses that are mostly used for artificial insemination in many countries including Spain. We study how parameters of in vitro sperm quality and characteristics of Murciano-Granadina dairy goats may affect the in vivo fertility obtained after artificial insemination with semen doses chilled at 4 °C. Moreover, this information was used for obtaining predictive models of the fertility. Sixty-three ejaculates from 13 males were used to prepare chilled doses for the insemination of 495 goats over 13 sessions. Fresh and chilled sperm were evaluated for motility and plasma membrane integrity with a computer-assisted sperm analysis system and flow cytometry, respectively. Fertility was determined at parturition, according to the kidding goats. Overall fertility was 59.6%. Pearson's correlation coefficients between in vivo fertility and quality variables of fresh sperm were not significant and were low (below 0.34 in absolute value) for chilled sperm. Females' characteristics had a low negative impact on fertility (correlation coefficients of -0.19 with age, -0.20 with parturitions and -0.11 with total milk yield obtained in the best lactation). Fixed and mixed logistic regression procedures were used trying to explain the fertility results. None of the models accurately predicted fertility, but the best models included the percentage of total motile sperm or average path velocity from fresh semen, age of the females and the session effect (uncontrolled environmental effects). These analyses showed that primiparous goats were 2.42 times more likely to get pregnant than goats that had kidded four or more times. Our field assay data on fertility in Murciano-Granadina dairy goats highlighted the importance of making quality controls of sperm, of choosing the doses presenting high percentages of motile sperm exhibiting regular trajectories and of selecting the youngest goats for AI, after their first kidding. Efforts should continue to obtain better predictive models for improving fertility in goat dairy herds.

Feasibility of selection for male contribution to embryo survival as a way of improving male reproductive performance and semen quality in rabbits.

The objective of this research was assess the male contribution to the number of implanted embryos (IE) and embryo survival (ES) estimated as the rate of variance due to male genetic and permanent environmental effects regarding total variance. In prolific species, the number of embryos and ES at early stages of gestation could be considered as fertility measurements because it indicates the number and rate of fertilized ova which are able to initiate the embryo development. Analyzed traits were ovulation rate (OR) estimated as the number of corpora lutea in both ovaries, IE estimated as the number of implantation sites, and ES calculated as IE/OR. A total of 1477 records from 900 females were used to analyze OR, whereas 1081 records from 855 females and 201 males were used to analyze IE and ES. The number of animals in the pedigree was 1107. The model included the systematic effects of year-season, parity order, lactation, and the random effects of additive genetics and permanent environmental effects due to the female (for OR) or both sexes (for IE and ES). Gibbs sampling was used to estimate posterior distributions of model parameters. The heritabilities of the male contribution to IE and ES were low (0.05 [0.01, 0.10] and 0.07 [0.02, 0.12]) but these estimates are probably biased downward since laparoscopy was only performed on those does that were pregnant at d 12 of gestation, instead of on all mated does. The genetic correlations between all analyzed traits and also between male and female genetic components of IE and ES were inaccurate, and it was not possible to draw any conclusion about them. The proportion of variation due to the male nonadditive genetic plus permanent environmental effects for IE and ES was almost negligible (0.027 [0.001, 0.058] and 0.031 [0.002, 0.068] for IE and ES, respectively), being the repeatability for male contribution on IE and ES around 8 and 10%, respectively.

Genetic selection for ovulation rate and litter size in rabbits: Estimation of genetic parameters and direct and correlated responses.

The aim of this work was to estimate direct and correlated responses in survival rates in an experiment of selection for ovulation rate (OR) and litter size (LS) in a line of rabbits (OR_LS). From generation 0 to 6 (first selection period), females were selected only for second gestation OR estimated by laparoscopy. From generation 7 to 13 (second selection period), a 2-stage selection for OR and LS was performed. In stage 1, females having the greatest OR at second gestation were selected. In stage 2, selection was for the greatest average LS of the first 2 parities of the females selected in stage 1. Total selection pressure in females was about 30%. The line had approximately 17 males and 75 females per generation. Traits recorded were OR estimated as the number of corpora lutea in both ovaries, number of implanted embryos (IE) estimated as the number of implantation sites, LS estimated as total number of rabbits born recorded at each parity, embryo survival (ES) estimated as IE/OR, fetal survival (FS) estimated as LS/IE, and prenatal survival (PS) estimated as LS/OR. Data were analyzed using Bayesian methodology. The estimated heritabilities of LS, OR, IE, ES, FS, and PS were 0.07, 0.21, 0.10, 0.07, 0.12, and 0.16, respectively. Direct and correlated responses from this study were estimated in each period of selection as the difference between the average genetic values of last and first generation. In the first selection period, OR increased 1.36 ova, but no correlated response was observed in LS due to a decrease on FS. Correlated responses for IE, ES, FS, and PS in the first selection period were 1.11, 0.00, -0.04, and -0.01, respectively. After 7 generations of 2-stage selection for OR and LS, OR increased 1.0 ova and response in LS was 0.9 kits. Correlated responses for IE, ES, FS, and PS in the second selection period were 1.14, 0.02, 0.02, and 0.07, respectively. Two-stage selection for OR and LS can be a promising procedure to improve LS in rabbits.

Selection for ovulation rate in rabbits: direct and correlated responses estimated with a cryopreserved control population.

The aim of this work was to evaluate the response in 10 generations of selection for ovulation rate in rabbits using a cryopreserved control population. Selection was based on the phenotypic value of ovulation rate estimated at d 12 of second gestation by laparoscopy. To produce the control population, embryos from 50 donor females and 18 males, belonging to the base generation of the line selected for ovulation rate, were recovered. A total of 467 embryos (72-h embryos) were vitrified and stored in liquid N(2) for 10 generations. The size of both populations was approximately 10 males and 50 females. The number of records used to analyze the different traits ranged from 99 to 340. Data were analyzed using Bayesian methodology. A difference between the selected and the control populations of 2.1 ova (highest posterior density interval (HPD(95%))[1.3, 2.9]) was observed in ovulation rate (OR), but it was not accompanied by a correlated response in litter size (LS; -0.3; HPD(95%) [-1.1, 0.5]). The number of implanted embryos (IE) increased with selection in 1.0 embryo (HPD(95%) [-0.6, 2.0]), but this increase was not relevant. Prenatal survival, embryonic survival, and fetal survival (FS) were calculated as LS/OR, IE/OR, and LS/IE, respectively. Prenatal survival was reduced with selection (-0.12; HPD(95%) [-0.20, -0.04]), basically because of a decrease in FS (-0.12; HPD(95%) [-0.19, -0.06]). Embryonic survival could have slightly decreased (-0.05; HPD(95%) [-0.12, 0.02]). In summary, comparison with a control population showed that ovulation rate in rabbits increased with selection without any correlated response in litter size, basically because of a decrease in fetal survival.

Selection for ovulation rate in rabbits: genetic parameters and correlated responses on survival rates.

The aim of this work was to evaluate the correlated responses on survival rates after 10 generations of selection for ovulation rate (OR). Selection was based on the phenotypic value of ovulation rate estimated at d 12 of second gestation by laparoscopy. Traits recorded were litter size (LS), estimated as total number of rabbits born per litter in up to 5 parities; OR, estimated as the number of corpora lutea in both ovaries; the number of implanted embryos (IE), estimated as the number of implantation sites; the number of right and left IE (RIE and LIE); ovulatory difference (OD), defined as the difference between the right and the left OR, expressed as an absolute value; implantatory difference (ID), defined as the difference between RIE and LIE, expressed as an absolute value; embryonic survival (ES), calculated as IE/OR; fetal survival (FS), calculated as LS/IE; prenatal survival (PS), calculated as LS/OR. A total of 1,081 records were used to analyze ES, and 770 were used to analyze FS and PS. The number of records used to analyze the other traits ranged from 1,079 for ID to 3,031 for LS. Data were analyzed using Bayesian methodology. Genetic parameters of OR, OD, and LS were estimated in a previous paper. Estimated heritabilities of IE, ID, ES, FS, and PS were 0.11, 0.03, 0.09, 0.24, and 0.14, respectively. Estimated repeatabilities of IE, ID, and ES were 0.22, 0.12, and 0.20. Estimated phenotypic correlations of OR with ES, FS, and PS were -0.07, -0.26, and -0.28, respectively. Their estimated genetic correlations with FS and PS were negative (probability of being negative 1.00 and 0.98, respectively). Nothing can be said about the sign of the genetic correlation between OR and ES. Ovulation rate was phenotypically uncorrelated with ID. Their estimated genetic correlation was positive (probability of being positive 0.91). The genetic correlation of ID with PS and LS was not accurately estimated. Phenotypic and genetic correlations between LS and survival rates were positive (probability of being positive 1.00). In 10 generations of selection, FS decreased around 1% per generation. No correlated response in ES was observed. In summary, the decrease in FS in rabbits selected for OR seemed to be responsible for the lack of correlated response observed in LS.

Selection for ovulation rate in rabbits: genetic parameters, direct response, and correlated response on litter size.

The aim of this work was to evaluate the response to 10 generations of selection for ovulation rate. Selection was based on the phenotypic value of ovulation rate, estimated at d 12 of the second gestation by laparoscopy. Selection pressure was approximately 30%. Line size was approximately 20 males and 80 females per generation. Traits recorded were ovulation rate at the second gestation, estimated by laparoscopy as the number of corpora lutea in both ovaries; ovulation rate at the last gestation, estimated postmortem; ovulation rate, analyzed as a single trait including ovulation rate at the second gestation and ovulation rate at the last gestation; right and left ovulation rates; ovulatory difference, estimated as the difference between the right and left ovulation rates; litter size, estimated as the total number of kits born and the number of kits born alive, both recorded at each parity. Totals of 1,477 and 3,031 records from 900 females were used to analyze ovulation rate and litter size, respectively, whereas 1,471 records were used to analyze ovulatory difference, right ovulation rate, and left ovulation rate. Data were analyzed using Bayesian methodology. Heritabilities of ovulation rate, litter size, number of kits born alive, right ovulation rate, left ovulation rate, and ovulatory difference were 0.16, 0.09, 0.08, 0.09, 0.04 and 0.03, respectively. Phenotypic correlations of ovulation rate with litter size, number of kits born alive, and ovulatory difference were 0.09, 0.01, and 0.14, respectively. Genetic correlations of ovulation rate with litter size and with number of kits born alive were estimated with low accuracy, and there was not much evidence for the sign of the correlation. The genetic correlation between ovulation rate and ovulatory difference was positive (P = 0.91). In 10 generations of selection, ovulation rate increased in 1.32 oocytes, with most of the response taking place in the right ovary (1.06 oocytes), but there was no correlated response on litter size (-0.15 kits). In summary, the direct response to selection for ovulation rate was relevant, but it did not modify litter size because of an increase in prenatal mortality.

In vivo development of vitrified rabbit embryos: effects on prenatal survival and placental development.

The aim of this work is to study the effect of the vitrification procedure on prenatal survival and on placental development at the end of gestation in rabbits (Oryctolagus cuniculus). One hundred eighty-one females were slaughtered at 72 h of gestation. Morphologically normal embryos recovered at 72 h of gestation were kept at room temperature until transfer or vitrification. Vitrified embryos (320 embryos) were transferred into a total of 24 does and fresh embryos (712 embryos) were transferred into a total of 43 does. Females were induced to ovulate 72 h before transfer when fresh embryos were transferred and 60 to 63 h before transfer when vitrified embryos were transferred. Each recipient doe received eight embryos into the left oviduct and eight embryos into the right oviduct. The number of implanted embryos was estimated by laparoscopy as number of implantation sites at Day 14 of gestation. Recipient females were slaughtered by stunning and exsanguination 25 d after the transfer, and fetuses were classified according to their status. Live fetuses and fetal and maternal placenta were weighed Pregnancy rate was defined as the total number of females having at least one live fetus at Day 28 of gestation divided by the total number of females. Prenatal survival was estimated as live fetuses at Day 28 of gestation divided by the number of transferred embryos. The pregnancy rate after transfer of vitrified embryos (92%) was similar to that achieved with fresh embryos (86%), but prenatal survival was lower for vitrified than for fresh embryos (53% vs. 34%). We did not find differences in embryo survival from 72 h to implantation. Transfer of vitrified embryos reduced fetal survival from implantation to Day 28 (57% vs. 82%). Differences in the number of live fetuses at Day 28 of gestation were mainly due to the higher fetal mortality observed soon after implantation in pregnancies resulting from the transfer of vitrified embryos. A higher percentage of decidual reactions and atrophic maternal placentas (27.5% vs. 8.3%) and also of atrophic fetal and maternal placentas (12.1% vs. 5.3%) were observed after transfer of vitrified embryos. Both treatments showed similar percentage of dead fetuses (3.3% vs. 4%). Maternal placenta of the fetuses from fresh embryos was 15% heavier than maternal placenta of fetuses from vitrified embryos.

Divergent selection for uterine capacity in rabbits. II. Correlated response in litter size and its components estimated with a cryopreserved control population.

Our objective was to evaluate the correlated responses to selection for litter size and its components after 10 generations of divergent selection for uterine capacity (UC). A total of 294 intact females from the 11th and 12th generations of divergent selection for high and low UC and from a cryopreserved control population was used (139, 112, and 43 females, respectively). Uterine capacity was assessed as litter size in unilaterally ovariectomized females. Traits recorded on females for up to five parities were litter size (LS) and number born alive (NBA). Laparoscopy was performed in all females at d 12 of their second parity, and the ovulation rate (OR) and number of implanted embryos (IE) were recorded in these females. Embryo survival (ES = IE/OR), fetal survival (FS = LS/IE), and prenatal survival (PS = LS/OR) were computed. Correlated responses in LS and in its components were inferred using Bayesian methods. Correlated responses in LS were asymmetric. The divergence between high and low lines was 2.35 kits, mainly because of a higher correlated response in the low line (1.88 kits). The lower LS in the low line was associated with a lower PS (control - low = 0.14), because of decreases in ES and FS.

Divergent selection for uterine capacity in rabbits. III. Responses in uterine capacity and its components estimated with a cryopreserved control population.

This work evaluated the response to 10 generations of divergent selection for uterine capacity (UC) in rabbits to determine whether this response was symmetric by contrasting both lines against a cryopreserved control population. Animals came from the 13th generation of an experiment of divergent selection for UC and from a cryopreserved control population. The two UC lines were divergently selected for 10 generations, and selection was relaxed from the 11th generation until the 13th generation. Uterine capacity was estimated as litter size (LS) in unilaterally ovariectomized (ULO) does. To create the control population, embryos from the base generation were vitrified and stored in liquid N2 for 10 generations. Data from 461 pregnancies produced by 134 ULO does were used: 62 does from the high UC line, 55 females from the low UC line, and 17 females from the control line. The following traits were analyzed: ovulation rate (OR); number of implanted embryos (IE); (UC), estimated as total number of rabbits born; number born alive (NBA); prenatal survival (PS), estimated as UC/OR; embryo survival (ES), estimated as IE/OR; and fetal survival (FS), estimated as UC/IE. Ovulation rate, IE, PS, ES, and FS were measured by laparoscopy only in the second parity. Uterine capacity and NBA were measured over four parities. Responses in UC and its components were estimated as differences between the selected lines and the control line using a Bayesian approach. Selection for UC led to differences of 1.01 kits between the high and low lines, but this response was asymmetric. No differences were found between the high and control lines (high - control = -0.08), whereas the low and control lines differed by 1.08 kits, with a probability of the difference being greater than zero of 0.98. Difference between the high and low lines and between the control and low lines was one-half of the difference reported for correlated response in LS in previous studies. No differences in OR were detected among lines. The control and low lines differed by 1.06 IE, with a probability of the difference being higher than zero of 0.84. Prenatal survival for the low line was less than that of the control line. In summary, selection for UC was asymmetric, which was mainly due to a correlated response in PS. Response in UC was one-half of the difference reported for correlated response in LS in previous studies.

The effect of divergent selection for uterine capacity on fetal and placental development at term in rabbits: maternal and embryonic genetic effects.

The aim of this work was to study the effects of the genotype of the dam, the embryo, or their interactions on prenatal growth by performing double-reciprocal embryo transfers between two lines of rabbits divergently selected for uterine capacity. Females from high (n = 53) and low (n = 48) lines were slaughtered at 72 h of gestation, and recovered embryos were transferred to the oviducts of recipient does from the high (n = 23) and low (n = 19) lines. Each recipient doe received eight embryos from the high line into one oviduct and eight embryos from the low line into the other. Recipient does were slaughtered on d 28 of gestation. The percentages of live fetuses at 28 d of gestation were 89.2 and 74% for high and low recipient lines, respectively. Length and weight of the empty uterine horn and weight of the full uterine horn were not affected by either the recipient or by donor line. Fetal weight was affected by the recipient line but not by the donor line. Fetuses gestated in high recipient does were 7% heavier (P < 0.10) than those in the low recipient does. There was a donor and a donor x recipient interaction effect on fetal placental weight. Fetal placental weight was heavier (7%, P < 0.01) for embryos from the low line. Embryos from the high line gestated in low-line uteri showed a lower fetal placenta weight than did low-line embryos gestated in high-line uteri and low-line uteri (P < 0.05). Linear regression coefficients of fetal weight at term on fetal placental weights differed (P < 0.05) for the high and low donors (4.33 +/- 0.28 and 3.41 +/- 0.29 respectively). A significant effect of the donor genotype on individual placental length was observed (P < 0.05), which might have resulted from a smaller individual placental length of low-line embryos gestated high-line uteri (P < 0.10). Neither donor nor recipient lines affected maternal placental weight or available space for fetuses. Fetuses and their fetal placentae were heavier when receiving more than four blood vessels than when receiving less than three blood vessels (13 and 17% respectively, P < 0.05). Neither recipient nor donor genotype affected the number of blood vessels arriving at each live fetus. Thus, fetal weight depends on the maternal genotype, whereas fetal placental weight depends on the embryo genotype in these two lines of rabbits divergently selected for uterine capacity.

The effect of divergent selection for uterine capacity on prenatal survival in rabbits: maternal and embryonic genetic effects.

The aim of this work was to determine whether prenatal survival depends on the genotype of the mother or of the embryo and to identify the critical periods for prenatal mortality in two lines of rabbits divergently selected by high (H) and low (L) uterine capacity. Does from H (n = 124) and L (n = 115) lines were slaughtered at 72 h of gestation. Embryos recovered at 72 h of gestation were transferred to the oviducts of recipient does from the H (n = 23) and L (n = 19) lines. Each recipient does received eight embryos from the H line into one oviduct and eight embryos from the L line into the other. Recipient does were slaughtered on d 28 of gestation. No differences were found between lines in the embryo recovery either in ovulation rate (OR) or in fertilization rate of ova recovered. Recovery rate was higher for the H line (0.80 vs. 0.72, P < 0.01). The number of embryos recovered, fitting ovulation rate as a covariate, was also higher for the H line (9.74 vs. 8.78, P < 0.05). The H line showed a more advanced embryonic stage of development, having a higher percentage of blastocysts (PB) and a lower percentage of compact morulae (PCM) (38% vs. 20%, P < 0.001 for PB, and 51% vs. 64%, P < 0.01 for PCM). The percentage of early morulae was low and similar in both lines. Neither donor nor recipient lines affected embryonic survival from 72 h to 7 d of gestation. Fetal survival was affected by the recipient line (P < 0.05). An interaction between donor and recipient was found. Embryos from the H donor line had a better fetal survival rate than embryos from the L donor line (P < 0.05) in H recipient females. Within L recipient females, embryos from H and L donor lines showed similar fetal survival. Fetal survival was divided into early (from d 7 to 17 of gestation) and late (from d 17 to 28 of gestation). The high recipient line showed a higher early fetal survival than the L recipient line (P < 0.05). The same effect was observed for late fetal survival, but the difference between H and L recipient lines was lower (P < 0.10). Thus, fetal survival depends mainly on the maternal genotype, and the embryo genotype only affects fetal survival when embryo transfer is performed to a favorable maternal environment. Selection for uterine capacity in rabbits leads to modification of early embryonic survival and of early and late fetal survival, but differences are higher for early than for late fetal survival.

Evaluation of electrical stunning methods using alternative frequencies in commercial rabbits.

The stages of recovery and spontaneous physical activity were evaluated using five methods of electrical stunning with variable voltage and frequency in an experiment with commercial rabbits (n=50). The recovery stages, evaluated by video recording, were: the tonic phase; the clonic phase; the corneal reflex; the resumption to rhythmic breathing, the response to painful stimuli and the hindleg posture. In a second rabbit sample (n=96) the muscular pH of the m. biceps femori and the pH of the m. longissimus dorsi were recorded. The levels of stunning were acceptable in all rabbits, with small variations among methods, especially with respect to the last stage of recovery. The period of insensibility before the appearance of the rhythmic breathing was an average of 30.14 ± 0.5 s long. There was no significant effect on meat pH at 24 h.