Reproductive response of crossbred Bos taurus × Bos indicus cows to biostimulation by pre-pubertal and pubertal teasers.

Although the beneficial effect of biostimulation on reproduction has been reported, the influence of selectivity and social factors on the response to biostimulation has not received sufficient research attention in both Bos indicus and Bos indicus influenced cattle. Furthermore, 'green and cheap' strategies to improve cattle reproduction are currently in demand while Bos indicus influenced cattle with inferior reproductive performance, and farmers with economic limitations are common in tropical zones. Hence, to assess the reproductive response of crossbred taurus × indicus cows to biostimulation by pre-pubertal (PPM) or pubertal (PM) teasers males, two trials of 2 years each were conducted. Trial 1 n = 187 cows (Year 1:85 cows exposed to PPM and Year 2:102 cows exposed to PM). Trial 2 n = 196 cows (Year 1:101 cows exposed to PPM and Year 2:95 cows exposed to PM). The effect of exposing cows to PPM and PM on the intervals calving to first service (ICFS), calving to conception (ICC) and economic cost of days open (ECDO) was analysed using Kruskal-Wallis ANOVA and the effect of exposing cows to PPM and to PM on reproductive status at 90 days (RS90) and proportion of cows requiring hormonal protocols (PRH) was compared using χ2 analysis. Both ICFS and ICC were shorter (p < .0001) for PM-exposed females (96.12 ± 4.1 and 110.93 ± 2.9 days; respectively) compared with those PPM-exposed (134.41 ± 3.3 and 135.64 ± 2.4 days; respectively). With RS90, more (p < .0001) PM-exposed cows (50.7%) were pregnant compared with PPM-exposed cows (16.1%). The PRH was greater (p < .0001) in PPM-exposed cows (79.0%) compared with PM-exposed (27.9%). The ECDO was less (p < .0001) in PM-exposed cows (US$ 142.9 ± 3.8) compared with PPM-exposed (US$ 176.3 ± 2.9). In conclusion, cows exposed to PM had shorter ICFS and ICC compared with cows exposed to PPM. More cows exposed to PM were pregnant after 90 days, and PRH was less than cows exposed to PPM. Cows exposed to PM had a reduced ECDO than those exposed to PPM.

Induction of fertile estrus without the use of steroid hormones in seasonally anestrous Suffolk ewes.

OBJECTIVE: To evaluate the efficacy of treatments based on GnRH, GnRH-PGF2α, and/or intense exposure to novel rams to induce fertile estrus without the use of steroid hormones in seasonally anestrous Suffolk ewes. METHODS: In the first experiment, ewes were treated with one injection of GnRH, two injections of GnRH administered 7 days apart, or a sequence of GnRH-PGF2α-GnRH. In the second experiment anestrous ewes were exposed, for 36 days starting on the day of weaning, to groups of four rams of three different breeds that were alternated every day. Besides exposure to the males, the ewes were injected with saline solution (ME group, n=20), with GnRH (ME-GnRH group, n=20) or with a sequence of GnRH-PGF2α-GnRH (ME-GPG group, n=20). The rams used for male-effect were fitted with aprons to prevent mating, and ewes detected in estrus were bred to selected fertile rams. Ovarian activity was monitored by progesterone determinations in both experiments. RESULTS: In the first experiment sustained induction of ovarian activity was not achieved and no ewe was detected in estrus. In the second experiment induction of sustained ovarian activity was achieved in all groups. Most of the ewes were detected in estrus, 76.7 % of the ewes were mated during a 36-d breeding period and 71.7 % of all the ewes became pregnant during that period. No significant differences between groups were found for any of these variables. However, estrus detection efficiency was higher in the ME-GnRH group than in the ME group (p&lt;0.05). CONCLUSION: An intense male-effect, that included the continuous presence and frequent alternation of several rams of different breeds, was sufficient to induce ovarian activity and fertile estrus in Suffolk ewes during the period of deep anestrus without the use of hormones, although addition of GnRH improved the efficiency of estrus detection.

Subtropical goats ovulate in response to the male effect after a prolonged treatment of artificial long days to stimulate their milk yield.

The objectives of this study were to determine (i) if in subtropical goats that gave birth during mid-December, the exposition to an artificial long-day photoperiod consisting in only 14 hr of light per day can increase the milk yield and (ii) to test whether these females can respond to the male effect at the end of the prolonged photoperiodic treatment. In experiment 1, 17 lactating goats were maintained under natural short days (control group), while another 22 goats were maintained under artificial long days (treated group) consisting in 14 hr light and 10 hr darkness starting at day 10 of lactation. The continuous exposition to an artificial long-day photoperiod produced an increase in the milk yield level during the first 110 days of lactation (time × treatment interaction; p = .01), while none of the milk components were modified due to the photoperiodic treatment (p > .05). In experiment 2, all control and treated anovulatory goats were submitted to the male effect using photostimulated males. All females showed oestrous behaviour within the first 10 days that were in contact with males (100% in both groups; p > .05). Thus, the latency to onset of oestrus did not differ between females from control (58.2 ± 3.0 hr) and treated (62 ± 4.6 hr) groups. Male exposition provoked ovulation independently if females were previously under long days or natural photoperiod (96 vs 100%, respectively; p = .79). It was concluded that exposure to 14 hr of light per day in subtropical goats that gave birth in late autumn stimulates milk yield without preventing the ovulation in response to the male effect at the end of the prolonged photoperiodic treatment.

Appetitive and Consummatory Sexual Behaviors of Rams Treated with Exogenous Testosterone and Exposed to Anestrus Dorper Ewes: Efficacy of the Male Effect.

We determine whether Dorper rams treated with testosterone in the sexual resting season trigger both appetitive and consummatory sexual behaviors and sexual and reproductive outcomes in anestrous ewes at 26° North during March-April. Dorper rams (n = 12, 2.5 years) were randomly divided prior to mating in two groups: GT (treated group; n = 6), treated with testosterone propionate (25 mg i.m. every 3 days × 3 weeks) and GC (control group; n = 6), treated with saline. Thereafter, adult anovulatory Dorper ewes (n = 61) were exposed to the GT-rams while the other group (n = 60) was exposed to the GC-rams. Ram's appetitive sexual behavior was similar (50 %) in both groups, yet the GT-rams exerted 100 % of the consummatory sexual behavior. Sexual and reproductive outcomes of the anestrus females exposed to both male groups considered two phases: 0-15 and 16-25 days from exposure to males. Involved variables were: ewes in estrus, ewes ovulating, duration of estrus, pregnant ewes, and lambed ewes. All the average variables favored those ewes exposed to GT-males: ewes in estrus: 85 versus 40 %; ewes ovulating: 80 versus 53 %; duration of estrus: 37 versus 25 h; pregnant ewes: 60 versus 31 %, and lambed ewes: 57 versus 31 %. Results confirm that adult Dorper rams treated with exogenous testosterone had improved consummatory sexual behavior and induced increased ovulation and pregnancy rates when exposed to anestrus ewes. This feasible and inexpensive testosterone-based protocol through the male effect also enhanced both sexual and reproductive outcomes in previously anestrous Dorper ewes during the natural out-of-season. Results are important to speed up reproductive performance of sheep during the natural anestrous season at this latitude, while also they embrace interesting outcomes from a comparative sexual behavior stand point.

Reproductive performance of postpartum ewes treated with insulin or progesterone hormones in association with ram effect.

The reproductive performance of postpartum Santa Inês (SI) and Morada Nova (MN) ewes treated with insulin or progesterone hormones in association with ram effect was evaluated. Ewes from SI (n = 69) and MN (n = 69) breeds were randomly allocated to three groups of each breed (T1-ram effect only; T2-ram effect + insulin; T3-ram effect + progesterone). Progesterone concentrations (ηg/ml; mean ± SD) before and after introduction of rams (n = 6) were 0.51 ± 0.22 and 3.78 ± 0.68 (T1), 0.65 ± 0.21 and 3.77 ± 0.78 (T2) and 0.52 ± 0.21 and 3.84 ± 0.84 (T3) in SI ewes and 0.74 ± 0.19 and 3.71 ± 0.56 (T1), 0.70 ± 0.21 and 3.79 ± 0.75 (T2) and 0.81 ± 0.14 and 3.87 ± 0.80 (T3) in MN ewes, respectively. Thus, lower progesterone concentrations were found before the introduction of rams (p < .05). After the introduction of rams, preovulatory peaks of LH (ηg/ml) occurred at 28 (T1), 44 (T2) and 48 (T3) hr in SI ewes and at 64 (T1), 40 (T2) and 44 (T3) hr in MN ewes. The mean number of ovulations was similar between groups (p > .05), was 1.3 ± 0.51 (T1), 1.5 ± 0.54 (T2) and 1.6 ± 0.51 (T3) in SI ewes and 1.3 ± 0.51 (T1), 1.6 ± 0.51 (T2) and 1.6 ± 0.51 (T3) in MN ewes. In conclusion, the ram effect alone is effective at inducing and synchronizing oestrus in sheep under postpartum anoestrus, irrespective of hormone treatments.

Reproductive outcomes of anestrous goats supplemented with spineless Opuntia megacantha Salm-Dyck protein-enriched cladodes and exposed to the male effect.

The possible influence of the "male effect" upon reproductive outcomes of adult anestrous goats under marginal rangeland conditions and supplemented with protein-enriched Opuntia megacantha Salm-Dyck was evaluated. Reproductive variables included: estrus percentage (EST, %), estrus latency (ESL, hours), ovulation percentage (OP, %), ovulation rate (OR, units), average largest follicle at ovulation (LFO, mm), largest corpus luteum (LCL, mm), embryo number (EBN, units), and embryo implantation percentage (EIP, %). During early May, anestrous mix-breed adult goats (Criollo x Alpine-Saanen-Nubian; n = 38, 26° N) were randomly distributed to (1) Control (CC; n = 12), (2), Non-enriched Opuntia (NEO; n = 14), and (3) Protein-enriched Opuntia (PEO; n = 12). Neither LW (P > 0.05) nor BCS (P > 0.05) or any of the evaluated ovarian variables differed (P > 0.05) among treatments; EST = 89.66%, ESL = 53.66 h, OP = 70.33%, OR = 1.07 units, LFO = 4.5 mm, LCL = 9.6 mm, EBN = 0.94 embryos, and EIP = 48.66%. Irrespective of nutritional supplementation regime, all goats denoted an increased response to the male effect just in the middle of the anestrous season and managed under marginal grazing conditions during the dry season (May to June; 26° N). The use of the male effect successfully invoked neurophysiological pathways to re-activate ovarian follicular and luteal pathways during the natural anestrous season in the female goat. Yet, such successful physiological scenario was not equally exerted to promote an increased embryo implantation rate; this issue claims further consideration. Therefore, it is essential to align not only the peri-conceptional but also the peri-implantation stages to the best suited environmental conditions in the rangeland, in order to increase both reproductive and economic efficiency while promoting sustainability in those rangeland-based marginal goat production systems.

Biostimulation and pheromones in livestock: A review.

This review examines aspects of the phenomenon of biostimulation in swine, goats, sheep, cattle and deer, to improve the collective knowledge and exploitation of its relevant mechanisms and effects in animal production. The long-term goal is to implement biostimulation strategies that benefit livestock reproduction and production while being both cost-effective and socially acceptable.

Group or individual housing does not reduce socio-sexual and reproductive responses in anestrous goats during the first contact with the photo-stimulated buck.

Herein, we examined the effect of the type of housing (individual vs group housing) of anestrus female goats on plasma cortisol concentrations and socio-sexual behaviors during first contact with photo-stimulated male goats and determined the impact on sexual and reproductive responses after 15 d of contact with photo-stimulated males. Therefore, after weaning, 10 females each were individually and group-housed, respectively. Ten bucks were rendered sexually active by exposure to artificially long days (16 h of light and 8 h of darkness per day) for 2.5 mo. During the naturally increasing photoperiod, 15-mo-old females were exposed and maintained with males. On day 0, during the first contact with males, individually- and group-housed females exhibited similar plasma cortisol concentrations (22.6 vs 27.4 ng/mL, respectively). Likewise, socio-sexual behaviors did not differ between examined groups on day 0. Moreover, the interval from first contact with males to the first estrus, short and normal ovulatory cycles, ovulation, fertility, and prolificacy rates were similar between differently housed females. Furthermore, mounting attempts did not differ between males in contact with either female group. In conclusion, individually- and group-housed anestrus females displayed elevated and similar plasma cortisol concentrations during first contact with photo-stimulated males and similar socio-sexual behaviors and reproductive responses when exposed to photo-stimulated males.

A Circular Economy Approach to Integrate Divergent Ruminant Production Systems: Using Dairy Cow Feed Leftovers to Enhance the Out-of-Season Reproductive Performance in Goats.

Based on a circular economy approach, we evaluated the possible effect of targeted supplementation with leftover feed from dairy cows (i.e., intensive system) on the reproductive performance of crossbred/rangeland goats (i.e., extensive system) in arid Northern Mexico. During the deep-anestrous season (i.e., March-April, 25° North), multiparous goats (n = 38) with a similar body weight (BW) and body condition score (BCS) were randomly assigned to two groups: (1) supplemented group (SG; n = 19; BCS: 1.76 ± 0.07; BW: 43.7 ± 1.8 kg), receiving 400 g goat d-1 of dairy-cow-feed leftovers prior to grazing; and (2) non-supplemented group (NS; n = 19; BCS: 1.76 ± 0.06; BW: 44.3 ± 2.5 kg). Both groups were directed to the rangeland for a period of ≈8 h. While the experimental period lasted 36 d, the experimental breeding considered 11 d (d0-d10). The anovulatory status of goats was ultrasonographically confirmed on days -20, -10, and -1 prior to male-female interaction. Previously, bucks were separated for 3 weeks from the experimental females and received exogenous testosterone every third day (i.e., 50 mg i.m.) prior to mating. With respect to the response variables, namely BW, BCS, blood glucose levels (BGLs), estrus induction (GIE, %), latency to estrus (LTE, h), estrus duration (ED, h), and luteal tissue volume (LTV, mm), no differences (p > 0.05) occurred between experimental groups. However, the response variables, namely goats ovulating (GO, %; 78.9 vs. 47.3), total number of corpuses luteum (TNCL, n; 27 vs. 13), ovulation rate (OR, n; 1.42 vs. 0.73), multiple ovulation (MO, %; 73.3 vs. 55.5), and pregnancy rate on d 36 (PRd36, %, 68.4 vs. 36.8), favored (p < 0.05) the SG over the NS goats. Our results demonstrate that connecting the circularity of two divergent ruminant production systems (i.e., cow-intensive and goat-extensive) by using dairy cows' feed leftovers as a targeted supplementation strategy in anestrous goats under a marginal-rangeland production system enhanced out-of-season reproductive outcomes (i.e., ovulation rate and pregnancy rate), thus benefiting marginal goat producers and their families.

Exposure of rams in sexual rest to sexually activated males in spring increases plasma LH and testosterone concentrations.

Eight stimulating rams, and twelve stimulated rams, were used to determine whether a similar endocrine response to the introduction of sexually active males in spring in a flock of ewes is observed in a flock of rams. The stimulating rams (n = 4) were induced into a sexually active state by exposure to 2 months of long days (16 h light/d) (15 December-15 February). At the end of the long-day period, rams were returned to the natural photoperiod. Control-stimulating rams (n = 4) were kept under the natural photoperiod. On April 20, stimulated rams were divided into 2 groups, and joined with activated (ACT; n = 6) or control stimulating rams (C; n = 6). On the day of ram introduction, stimulated rams were blood sampled for 8 h at 20-min intervals, from 4 h before to 4 h after ram introduction, and next day from 24 to 28 h after ram introduction, and analyzed for plasma LH concentrations, and 10, 20 and 30 days after ram introduction to measure plasma testosterone levels. Mean (±SEM) plasma LH concentrations (ng/ml) of stimulated rams were similar during the 4 h before stimulating-ram introduction (ACT: 0.59 ± 0.03; C: 0.53 ± 0.04; P > 0.05). The introduction of the photoperiod-treated stimulating rams increased LH concentrations of stimulated rams during the 4 h after their introduction (1.14 ± 0.37) compared with the C group (0.51 ± 0.03; P < 0.05), especially during the first hour (ACT: 0.93 ± 0.16; C: 0.49 ± 0.03; P < 0.05), and during the blood sampling period 24-28 h after ram introduction (0.75 ± 0.07 vs. 0.58 ± 0.04; P < 0.05). Before the introduction of stimulating rams, the LH pulse frequencies and amplitudes did not differ between groups; however, LH pulsatility was higher at 4 h (0.58 ± 0.11 pulses/h; P < 0.05), and had trend to be higher 24 h (0.50 ± 0.06) (P = 0.10) after the introduction of the photoperiod-treated stimulating rams compared with the control-stimulating rams (0.29 ± 0.08 and 0.29 ± 0.10, respectively). As for LH pulses, there was an effect of group (P < 0.05) on LH amplitude, which presented a trend to be higher in ACT rams 4 h after ram introduction (1.68 ± 0.30; P < 0.10) and higher 24 h (1.07 ± 0.08; P < 0.05) after ram introduction, compared with LH amplitudes of C rams (0.71 ± 0.06 and 0.82 ± 0.07, respectively). Plasma testosterone concentrations of rams exposed to photoperiod-treated activated rams were higher than those of rams exposed to control-stimulating rams, at 4 h, 20 and 30 days after ram introduction (P < 0.05). In conclusion, sexually active rams in spring are able to stimulate LH and testosterone secretion of other rams in sexual rest, a phenomenon we called "ram-to-ram effect".

The High Testosterone Concentrations of the Bucks Used in the "Male Effect" Is Not a Prerequisite for Obtaining High Ovarian Activity in Goats from Mediterranean Latitudes.

Two experiments were carried out. Firstly, 54 anestrous females were placed in contact with photostimulated males (Photo; n = 27) or with no photostimulated males (Natural; n = 27). Moreover, a group of bucks treated with artificial photoperiod and a group of bucks subjected to natural photoperiod without contact with females was used (Photo Isolated and Natural Isolated, respectively). In the Natural groups, the testosterone concentrations were similar except for three days after the introduction of the bucks to the does (19.72 ± 4.11 vs. 2.05 ± 0.25 ng/mL for Natural and Natural isolated bucks, respectively, p < 0.05). However, no differences were observed in the Photo groups during the entire experiment. The percentage of females showing estrous was higher in the group of females in contact with photostimulated bucks (96 vs. 74%, respectively, p < 0.05). In the second experiment, a GnRH agonist, deslorelin, was used to regulate the testosterone concentrations of the bucks. Seventy anestrous females were divided into five groups depending on the treatment received by the bucks to which they were exposed: photostimulated bucks (Photo group, n = 14); photostimulated bucks but treated with the agonist at the onset of the photoperiod treatment (Photo-Ago Long group, n = 13); photostimulated bucks but treated with the agonist at the end of the photoperiod treatment (Photo-Ago Short group, n = 15); bucks receiving no photostimulation but treated with the agonist at the end of the photoperiod treatment period (Natural-Ago Short group, n = 13) and bucks receiving no photostimulation nor agonist (Natural group, n = 15). The agonist treatment increased testosterone concentrations after the injection, which remained high for the entire experiment (p < 0.05). Six days after the introduction of the bucks to the does, the testosterone concentrations increased only in the Natural group reaching similar concentrations to the other groups (12.17 ± 6.55, 16.69 ± 4.53, 8.70 ± 0.61, 11.03 ± 1.45 and 14.42 ± 3.64 ng/mL for Photo, Photo-Ago Long, Photo-Ago Short, Natural-Ago Short and Natural bucks, respectively, p > 0.05). No differences in reproductive parameters were observed (p > 0.05). These results demonstrate that, at Mediterranean latitudes, anestrous females can stimulate the testosterone concentrations of bucks after a period of isolation. The high testosterone concentrations are not a prerequisite for an adequate response to the male effect.

High body condition score combined with a reduced lambing to ram introduction interval improves the short-term ovarian response of milking Lacaune ewes to the male effect.

The male effect is an effective natural technique to induce off-season ovulation and ultimately mating or artificial insemination in small ruminants. It constitutes an alternative to hormonal treatments in conventional breeding systems and, to shift and organise the yearly production cycle, is currently the only solution complying with European organic standards. However, its associated performances are still heterogeneous, both in terms of the global response and the extent of reproductive synchronisation of the females, due to complex interactions with environmental factors that limit its use on commercial farms. This study was carried out on a French organic farm under commercial conditions to investigate, in the field and across five consecutive years, the main parameters affecting the early ovarian response to a ram effect on Lacaune dairy ewes. While the within-year binary logistic regressions yielded contrasting results, the cross-year mixed-effect binary logistic regression models clearly showed that parameters associated with the nutritional state of the animals have a profound influence on the ovarian response of the ewes. Indeed, the probabilities of a spontaneous resumption of ovarian activity before the ram effect and of an early ovarian response to the ram effect were positively associated with the body condition score, total milk production and the age of the animals, while being negatively associated with the milk production level at the 3rd milk recording. The probabilities of a spontaneous resumption of ovarian activity before the ram effect were positively associated with the interval between lambing and the introduction of the rams. Altogether, these results indicate that the ovarian performances in response to a male effect follow a bell-curve pattern with optimal performances depending upon a complex combination between photoperiodic and nutritional cues. Regarding these latter, this study highlights the major contribution of body reserves and energy balance dynamics.

Biostimulation with the ram effect increases the follicle recruitment, ovulatory diameter, and embryo viability rate in superovulated ewes.

The aim of this study was to compare the preovulatory follicular development and superovulatory outcomes in superovulated ewes which were either stimulated or not stimulated by placement with rams. The treatment regimen to super-stimulate ovarian follicular development was imposed to 28 ewes on "Day 0", from which 14 were stimulated with active rams for 48 h, starting at the time of the fifth FSH dose, with the ram being removed from the pen with the ewes and replaced which other ram every 12 h, using four different rams (group GRE). The other 14 ewes remained isolated from rams throughout the protocol (group GCON). All ewes were administered 133 mg of FSH, into six doses in decreasing quantities, every 12 h. The follicular development and number of ovulations were determined using ultrasonography. Biostimulation resulted in an increased number of large follicles, follicle diameter, and embryo viability rate (viable embryos/recovered structures∗100) was greater in ewes of the GRE than GCON group. The number of corpora lutea, follicular cysts, recovered structures, viable embryos, and degenerated and unfertilized structures was similar in ewes of the GRE and GCON group. Structures were recovered from more GRE than GCON ewes. In conclusion, biostimulation with rams during the last phase of the treatment regimen to induce superovulation enhanced the follicular growth and increased the embryo viability rate in ewes.

Reproductive Resumption in Winter and Spring Related to MTNR1A Gene Polymorphisms in Sarda Sheep.

The aim of the present research was to evaluate the association between the SNPs rs430181568 and rs407388227 located in the MTNR1A gene with the reproductive recovery of Sarda sheep in different months of ram introduction in the flock (February, March, April and May). In order to address this research gap, we selected two farms, each of which consisted of approximately 1000 animals; a total of 800 ewes (400 for each farm) were genotyped for the two single nucleotide polymorphisms rs430181568 and rs407388227 located in the exon 2 of the MTNR1A. These SNPs are completely linked; thus, each genotype of rs430181568 corresponded to the same genotype for rs407388227. Among the genotyped animals, 240 individuals were selected and divided into four homogeneous groups (A, B, C and D) of 60 subjects, each group based on age (range 3−6 years old), body condition score (BCS) (range 2.0−4.0) and genotype (20 ewes carrying CC/CC, 20 CT/CT and 20 TT/TT genotype). The dates of the ram introduction in each group were 15 February, 15 March, 15 April and 15 May, respectively. In all groups, the lambing date and the number of lambs born from 150 to 220 days after the ram introduction were recorded. In all the groups, the genotypes CC/CC and CT/CT of the polymorphism (rs430181568 and rs407388227) showed the greatest fertility (the ratio between the number of lambed ewes and the ewes exposed to the rams) (p < 0.01) and the shortest distance between ram introduction to lambing (p < 0.01), compared with the TT/TT genotype. In conclusion, we determined that the polymorphisms rs430181568 and rs407388227 were associated with reproductive recovery, after ram introduction, even in February and March, months subsequent to the photorefractoriness period.

Sexually active male goats stimulate the endocrine and sexual activities of other males in seasonal sexual rest through the "buck-to-buck effect".

Male goats rendered sexually active by exposure to a photoperiodic treatment are more efficient than untreated goats in stimulating LH secretion and ovulation in seasonally anestrous goats. This phenomenon is called the "male effect." Here, we determined whether sexually active bucks are able to stimulate the endocrine and sexual activities of other bucks in seasonal sexual rest through the phenomenon that we called the "buck-to-buck effect." We used bucks rendered sexually active (SA) during sexual rest by exposure to 2.5 mo of artificial long days (16 h of light per d) and untreated, sexually inactive (SI) bucks. In Experiment 1, we determined the short-term (21 d) LH and testosterone responses of sexually inactive bucks joined with a SA or SI buck. In Experiment 2, we determined the long-term (60 d) testosterone and sexual behavior responses of sexually inactive bucks joined with 2 SA or SI bucks. In Experiment 3, we determined the efficacy of bucks initially exposed to the buck-to-buck effect, the SABB bucks, to thereafter induce a "classical" male effect in seasonally anestrous goats. In Experiments 1 and 2, there was an interaction between time and groups in LH and testosterone plasma concentrations (P < 0.01). In Experiment 1, plasma LH concentrations were greater in bucks joined with a SA buck than in those joined with an SI buck (P < 0.05). In Experiments 1 and 2, testosterone concentrations were greater in bucks joined with SA bucks than in those joined with SI bucks (P < 0.05). In addition, in Experiment 2, the sexually inactive bucks joined with SA bucks displayed more nudging than those joined with SI bucks (P < 0.001). In Experiment 3, kidding rates did not differ between females joined with SA (34 of 40: 85%) or SABB bucks (32 of 40: 80%; P > 0.05). We concluded that the endocrine and sexual activities of bucks during sexual rest can be stimulated by SA bucks. In addition, SABB bucks are able to stimulate the reproductive activity of seasonally anestrous goats.

Variation of ewe olfactory secretome during a ram effect.

Introduction: Under temperate latitudes, reproduction in Ovis aries displays a marked seasonality, governed by the photoperiod. In natural conditions, the transition between sexual rest and sexual activity in both sexes is induced by the decrease of day light. Meanwhile, specific odors emitted by a sexually active male are able to reactivate the gonadotropic axis of anovulatory ewes. This physiological effect is called "male effect", precisely ram effect in the ovine species. We have previously shown that the secreted proteins, namely Olfactory Binding Proteins (OBP), contained in the nasal mucus constitute the olfactory secretome (OS), the composition of which is determined by the status of oestrus cycle of females and differs between sexual rest and sexual activity periods. The objective of this study was to test the hypothesis that exposure to sexually active male can also modify the composition of ewes olfactory secretome during a male effect, as well as hormones produced by the reactivation of the oestrus cycle in sexual activity period under natural conditions. Methods: We have set up a new non-invasive protocol of nasal mucus sampling and collected it from 12 ewes at different times during a ram effect. We analyzed the composition of their olfactory secretome by proteomics, mainly SDS-PAGE and MALDI-TOF mass spectrometry. As post-translational modifications of OBPs were a hallmark of ewes' sexual activity period, we were looking for glycosylation by western-blot and mass spectrometry. Results: The efficiency of male effect was low in stimulated ewes as only 3 females displayed elevated progesterone levels in their blood. Besides, half of control ewes (non-stimulated ones) were cycled. We noticed a common OS profile in ewes in anoestrus, versus OS of cycled ones. A very clear and important result was the apparition of O-GlcNAcylation, previously detected only in sexual activity, after only 30 min of male introduction into the flock. Discussion: This exploratory study paves the way for further experiments with larger flock to confirm and reinforce these results, and for eventually exploiting the nasal mucus as an indicator of females' receptivity to male odors.

The use of photoperiod-treated bucks to induce a "male effect" does not compensate for the negative effects of nutritional restriction of the females in Mediterranean goats.

This work examined the effect of acute nutritional restriction or supplementation one week before male introduction on the reproductive performances of the "male effect" when using photostimulated or control males in goats. On 22 March, 84 anoestrous does were placed with photostimulated bucks or with bucks which had received no treatments. One week before male introduction, the females were provided with different nutritional regimes: Supplemented, restricted or control females. The non-esterified fatty acids (NEFAs) and Insulin Growth Factor-1 (IGF-1) concentrations were measured in the same samples. Fecundity, fertility, prolificacy and productivity were also determined. No interaction between both sources of variation was observed in any of the reproductive variables studied. Treatment of the bucks increased the percentage of females expressing behavioural oestrous associated with ovulation (71% vs 90% for Natural and Photo groups, respectively, P < 0.05). The Supplemented females showed higher ovulation rate than Restricted females (1.77 ± 0.13 vs 1.05 ± 0.05, P < 0.001), fecundity (71% vs 43%, P < 0.05); fertility (76% vs 29%, P < 0.05) and productivity (1.00 ± 0.15 vs 0.29 ± 0.11 kids per female, P < 0.01). In the Supplemented females, the higher reproductive results could be due to the lower NEFAs and higher IGF-1 concentrations at ovulation and at the time of oestrus compared to the Restricted females. Thus, the present experiment results demonstrate that nutrition is an important factor in the response to the "male effect" at Mediterranean latitudes, and its negative effect cannot be counterbalanced by using photostimulated bucks.

Nutritional supplementation improves the sexual response of bucks exposed to long days in semi-extensive management and their ability to stimulate reproduction in goats.

In confined management systems, well-nourished bucks rendered sexually active by exposure to long days are efficient in fertilizing out-of-season goats. However, underfeeding is common in semi-extensive management systems and may reduce the reproductive efficiency of bucks. The objective of the present study was to determine whether nutritional supplementation improved the sexual activity of bucks submitted to long days in semi-extensive management systems and their ability to stimulate the reproduction of goats in semi-extensive or confined conditions. In experiment 1, three groups of bucks were placed in different flocks and grazed daily with females for 7 h. Each day after grazing, males were separated from females and moved into open pens. One group did not receive any treatment (control group; n = 6). Two other groups were submitted to artificially long days from 15 November to 15 January. From 16 January, one group did not receive nutritional supplementation (long-day group; n = 5), whereas bucks from the other group each received 600 g of a commercial concentrate (long-day+supplementation group; n = 5). The fourth group was kept in confined conditions, exposed to long days and fed alfalfa hay (long-day confined group; n = 6). On 26 March, anovulatory goats from other flocks were assigned to four groups (n = 27 each) and confined separately in open pens. Three bucks of each group were housed with the females. Pregnancy rates were greater in the goats housed with the long-day group than those housed with the control group (P < 0.01). However, pregnancy rates did not differ between the long-day confined group (89%) and long-day+supplementation group (70%; P = 0.09), but these rates were greater than those from the long-day (37%) and control groups (0%; P < 0.05). In experiment 2, two groups of males (n = 3 each) were incorporated into two flocks under semi-extensive management and grazed daily with females for 7 h. One group of males did not receive any treatment (control group). The other group was submitted to long days and nutritional supplementation as in experiment 1 (long-day+supplementation group). Males remained with females during the whole study. The pregnancy rate was greater in the goats joined by males of the long-day+supplementation group (78%) than in those from the control group (0%; P < 0.001). We conclude that long days and nutritional supplementation improve the ability of bucks kept in semi-extensive management to stimulate reproduction of out-of-season goats in confined or semi-extensive management systems.

Steroidome and metabolome analysis in gilt saliva to identify potential biomarkers of boar effect receptivity.

Optimal management of gilt reproduction requires oestrus synchronization. Hormonal treatments are used for this purpose, but there is a growing demand for non-hormonal alternatives, especially in organic farms. The boar effect is an important alternative opportunity to induce and synchronize oestrus without hormones. Before puberty, gilts exhibit a 'waiting period' during which boar exposure could induce and synchronize the first ovulation. We searched for salivary biomarkers of this period of boar effect receptivity to improve detection of the gilts to stimulate with the perspective of enhancing the efficacy of the boar effect. Saliva samples were collected from 30 Large-White×Landrace crossbred gilts between 140 and 175 days of age. Gilts were exposed twice a day to a boar and subjected to oestrus detection from 150 to 175 days of age. Among the 30 gilts, 10 were detected in oestrus 4 to 7 days after the first introduction of the boar and were considered receptive to the boar effect, 14 were detected in oestrus more than 8 days after first boar contact, and six did not show oestrus and were considered non-receptive. Saliva samples from six receptive and six non-receptive gilts were analyzed for steroidome and for metabolome using gas chromatography coupled to tandem mass spectrometry and 1H nuclear magnetic resonance spectroscopy, respectively. Four saliva samples per gilt were analyzed: 25 days and 11 days before boar introduction, the day of boar introduction, 3 days later for receptive gilts or 7 days later for non-receptive gilts. Twenty-nine steroids and 31 metabolites were detected in gilt saliva. Salivary concentrations of six steroids and three metabolites were significantly different between receptive and non-receptive gilts: progesterone and glycolate 25 days before boar introduction, 3α5ß20α- and 3ß5α20ß-hexahydroprogesterone, dehydroepiandrosterone, androstenediol, succinate, and butyrate 11 days before boar introduction, and 3ß5α-tetrahydroprogesterone on the day of boar introduction. Thus, nine potential salivary biomarkers of boar effect receptivity were identified in our experimental conditions. Further studies with higher numbers of gilts and salivary sampling points are necessary to ascertain their reliability.

Photoperiod-Treatment in Mediterranean Bucks Can Improve the Reproductive Performance of the Male Effect Depending on the Extent of Their Seasonality.

This study examines the effectiveness of the photoperiod treatment (extra light for 88 days) to render the bucks sexually active during the seasonal anoestrous in order to induce a male effect, depending on the reproductive seasonality of the breed of the bucks used. In April, 57 anoestrous Blanca Andaluza does were distributed into four groups with three males each: 13 were exposed to control Murciano-Granadina bucks (lower seasonality); 15 were exposed to photostimulated Murciano-Granadina bucks; 14 were exposed to control Blanca Andaluza bucks (higher seasonality), and 15 were exposed to photostimulated Blanca Andaluza bucks. After male introduction, the sexual behaviour of the bucks was assessed, and harness marks recorded doe oestrous behaviour. Ovulation was confirmed from plasma progesterone, and the ovulation rate was assessed by transrectal ultrasonography. Fecundity, fertility, prolificacy and productivity were also determined. All of the does in all of the groups showed ovulation. Interaction between both sources of variation was observed: the percentage of females showing oestrous (p < 0.01) and productivity (p < 0.05) was the lowest in the Blanca Andaluza control group (50% and 0.36 ± 0.17 goat kids born/female, respectively). In conclusion, photoperiod-treated bucks efficiently induce a male effect, but photostimulation could be more necessary for breeds with deep seasonality.