77 USE OF SEMINAL PLASMA TO IMPROVE REPRODUCTIVE PERFORMANCE IN ALPACAS (VICUGNA PACOS) FOLLOWING NATURAL MOUNTING

2016 ◽  
Vol 28 (2) ◽  
pp. 168
Author(s):  
W. Huanca ◽  
J. Turin ◽  
C. Mamani ◽  
R. Sanchez ◽  
W. F. Huanca ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Reproductive Performance ◽  
Control Experiment ◽  
Conception Rate ◽  
Gnrh Analogue ◽  
Dominant Follicle ◽  
Linear Transducer ◽  
Vicugna Pacos ◽  
Buserelin Acetate ◽  
Natural Mating

Alpacas are classified as induced ovulators, and an external stimulus is required for the occurrence of ovulation. The seminal plasma (SP) of camelids contains a protein identified as β nerve growth factor with the capacity to induce ovulation and corpus luteum formation. Alpacas have a poor reproductive efficiency and high embryo mortality rate. A study was designed to evaluate the use of SP to improve reproductive performance post natural mating (Exp. 1) and with different mounting time (MT; Exp. 2). Experiment 1: Nonpregnant alpacas (n = 117) exhibiting a dominant follicle = 7 mm, detected by transrectal ultrasonography, were bred by natural mating and then assigned randomly to 1 of 3 treatments: T1 (n = 40), 1 mL of SP IM; T2 (n = 39), 1 mL of GnRH analogue (0.0042 mg of buserelin acetate), IM; and T3 (n = 38), control. Experiment 2: Nonpregnant alpacas (n = 180) exhibiting a follicle = 7 mm were assigned randomly to the following treatments: T1 (n = 30): MT 5 min; T2 (n = 30): MT 5 min + 1 mL of SP; T3 (n = 30): MT 10 min; T4 (n = 30): MT 10 min + 1 mL of SP; T5 (n = 30): MT > 15 min; T6 (n = 30): MT > 15 min + 1 mL of SP. The same male was used for mating females in T1 and T2, T3 and T4, and T5 and T6, respectively. Animals were evaluated by ultrasound with an Aloka SSD 500 (Aloka, Tokyo, Japan) and 5.0-MHz linear transducer on Day 25 for pregnancy diagnosis. In Exp. 1 the conception rate was 67.5, 51.3, and 55.3% for T1, T2, and T3, respectively (P > 0.05). In Exp. 2 the conception rate was 46.2, 67.9, 57.1, 73.3, 61.8, and 72.7% for T1, T2, T3, T4, T5, and T6 (P < 0.05) and was significantly different between T1 v. T2 and T3 v. T4. The results suggest that the use of SP could be one method to improve reproductive performance in alpacas. Research was supported by Project N° 405-PNICP-PIAP-UNMSM.

165 EFFECT OF INJECTION OF SEMINAL PLASMA ON OVULATION RATE, CORPUS LUTEUM DEVELOPMENT, AND SENSITIVITY TO PROSTAGLANDIN IN ALPACAS (VICUGNA PACOS)

2015 ◽  
Vol 27 (1) ◽  
pp. 173
Author(s):  
W. F. Huanca ◽  
C. Mamani ◽  
W. Huanca
Keyword(s):  
Corpus Luteum ◽  
Seminal Plasma ◽  
Saline Solution ◽  
Ovulation Rate ◽  
Similar Response ◽  
Linear Transducer ◽  
Vicugna Pacos ◽  
Buserelin Acetate ◽  
T1 And T2 ◽  
Control Application

The seminal plasma (SP) of camelids contains a protein identified as β Nerve Growth Factor with capacity of induced ovulation and develop a corpus luteum. A study was designed to evaluate the effect of application of SP on the interval of time from injection of stimulus to the ovulation and corpus luteum (CL) size (Experiment 1, n = 24) and on the sensitivity of CL to the injection of prostaglandin (196 µg of tiaprost) (PG) at different periods from ovulation (Experiment 2, n = 86). Exp. 1: Adult female alpacas with presence of a follicle ≥7 mm were assigned to the application of 1 mL of SP via IM (T: n = 12) or application of 50 µg of acetate of busereline IM (T2: n = 12). Exp. 2: Alpacas with presence of a follicle ≥7 mm were induced to ovulation with 50 µg of acetate of busereline or 1 mL IM of SP. Animals were evaluated by ultrasound to confirm the ovulation and were assigned to the following treatment: T1 (n = 8): SP + PG Day 4; T2 (n = 8): buserelin acetate + PG Day 4; T3 (n = 8): SP + PG Day 5; T4 (n = 8): buserelin acetate + PG Day 5; T5 (n = 8): SP + PG Day 6; T6 (n = 8): buserelin acetate + PG Day 6; T7 (n = 8): SP + PG Day 7; T8 (n = 8): buserelin acetate + PG Day 7; T9 (n = 8): SP + PG Day 8; T10 (n = 8): buserelin acetate + PG Day 8 and T11 (n = 6) (Control): Application of 1 mL of saline solution. The animals were evaluated by ultrasound with an Aloka SSD500 (Aloka, Tokyo, Japan) and 7.5-MHz linear transducer each 2 h (Exp. 1) and each 12 h (Exp. 2) after of application of PG. In Exp. 1, the ovulation rate was 95.7% to T1 and T2 and an interval of time between injection of stimulus and ovulation was 27.4 ± 2.5 h and 26.8 ± 1.8 to T1 and T2, respectively. In Exp. 2, luteolysis was 0.0%, 0.0%, 25.0%, 0.0%, 100.0%, 100.0%, 100.0%, 100.0%, 100.0%, 100.0%, and 0.0% for the treatments T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, and T11 respectively. The results suggest that no differences exist between in the ovulation rate and interval to the ovulation between the application of buserelin acetate or SP and that the CL was sensible at Day 5 to the prostaglandin respect SP and with similar response to the sensibility of CL from Day 6 to Day 8.

108 Use of seminal plasma as ovulation inductor in alpacas (Vicugna pacos) embryo recipient and its effect on pregnancy rate

2020 ◽  
Vol 32 (2) ◽  
pp. 181
Author(s):  
W. Huanca ◽  
K. Garcia ◽  
W. F. Huanca ◽  
A. Cordero ◽  
J. Malaga
Keyword(s):  
Pregnancy Rate ◽  
Seminal Plasma ◽  
Body Condition Score ◽  
Gnrh Analogue ◽  
Vicugna Pacos ◽  
Buserelin Acetate ◽  
Chi Squared ◽  
Condition Score ◽  
Equine Chorionic Gonadotropin ◽  
Phosphate Buffered Saline

Alpacas, like other camelids, are classified as induced ovulators because an external stimulus is required for the occurrence of ovulation. Recent studies have identified the β-nerve growth factor (β-NGF), a protein present in the seminal plasma (SP), as responsible for inducing ovulation in this species and having potent luteotropic function. We speculate that SP can be used in reproductive biotechnologies, such as embryo transfer (ET), to increase the number of genetically superior animals produced in breeding programs. The objective of this study was to evaluate the effect of inducing ovulation with SP or a gonadotrophin-releasing hormone (GnRH) analogue on pregnancy rate in recipients of an alpaca ET program. Semen from 5 adult male alpacas was collected with an artificial vagina and diluted 1:1 (v/v) with phosphate-buffered saline (PBS; Gibco-BRL). The diluted semen was centrifuged for 30min at 1200g, and the supernatant or SP was decanted and examined for absence of sperm. Then, the SP was centrifuged again for 20min at 1200×g. The SP was stored at −80°C until the use. Female alpacas (n=38; 6-8 years) with a body condition score of 2.5-3.5 (scale 1-5) were used for the experiment. Animals were evaluated daily by transrectal ultrasonography to determine the presence of a dominant follicle ≥7mm and randomly assigned to one of two groups: (1) GnRH (n=20), 0.04mg of Buserelin acetate IM, or (2) SP (n=18): 1.0mL of seminal plasma IM. Seven days after GnRH or SP treatment (Day 0) the recipients received a fresh embryo. Alpaca donors (n=18) were given GnRH (0.04mg IM) and treated 36h later with 700IU of equine chorionic gonadotropin (ECG). Donors were mated with fertile males 5 days after ECG (Day 0 of recipients), and embryos were recovered 7 days later. Embryos of similar quality were assigned to both groups and transferred nonsurgically to the uterine horn ipsilateral to the corpus luteum. Ultrasonography examinations were performed on Day 2 to confirm ovulation and Day 25 to determine pregnancy in all of the recipients. Data was analysed by chi-squared test. Ovulation rate was not different between groups (100% each). Pregnancy rate was 45% (9 out of 20) and 44% (8 of 18) in GnRH and SP groups, respectively (P=0.77). In conclusion, SP was effective to induce ovulation in alpacas and was able to produce a pregnancy rate similar to that of GnRH as an ovulation-inducing treatment. Thus, SP can be used as an alternative for ET programs in alpacas. Research was funded by the project Role of Seminal Plasma in Reproductive Physiology and Application of Biotechnologies in Camelids (149-2017-CIENCIACTIVA).

82 CRYOPRESERVATION OF BOAR EPIDIDYMAL SPERMATOZOA; ADDITION OF SEMINAL PLASMA TO THAWING SOLUTION IMPROVES REPRODUCTIVE PERFORMANCE BY ARTIFICIAL INSEMINATION

2011 ◽  
Vol 23 (1) ◽  
pp. 146
Author(s):  
T. Okazaki ◽  
T. Akiyoshi ◽  
M. Kan ◽  
H. Teshima ◽  
M. Shimada
Keyword(s):  
Litter Size ◽  
Seminal Plasma ◽  
Reproductive Performance ◽  
Artificial Insemination ◽  
Conception Rate ◽  
Maximum Effect ◽  
Dependent Manner ◽  
Epididymal Sperm ◽  
Epididymal Spermatozoa

Epididymal spermatozoa are one of the available male germ cells for cryopreservation. It has been reported that frozen–thawed porcine epididymal spermatozoa have a high fertilization competence in vitro as compared with that in ejaculated one. However, there is little information about reproductive performance, such as conception rate or litter size, after artificial insemination (AI) using frozen–thawed epididymal spermatozoa. Recently, we demonstrated that the addition of seminal plasma to thawing solution improves membrane and acrosomal integrity, and enhanced both in vivo and in vitro fertilizing activity of frozen–thawed ejaculated spermatozoa. Moreover, the injection of seminal plasma to uterus with frozen–thawed spermatozoa significantly increased the number of implantation site (Okazaki et al. 2009 Theriogenology 71, 491–498). Thus, to apply those positive functions of seminal plasma to AI using frozen–thawed epididymal sperm, in this study, we added seminal plasma to thawing solution and then analysed the sperm functions including AI test using frozen–thawed epididymal spermatozoa. Epididymal spermatozoa collected by flushing caudal epididymis were frozen as described in our previous study (Okazaki et al. 2009). Frozen-spermatozoa were thawed in Modena solution with or without different percentages of seminal plasma. Protein tyrosine phosphorylation as a marker of capacitation was detected by western blotting. To examine the reproductive performance, the sows of natural oestrus were artificially inseminated two times (5 × 109 50 mL–1 per injection). When the frozen–thawed ejaculated or epididymal sperm was incubated up to 6 h, the motility of epididymal sperm was significantly higher than that of ejaculated sperm (19.6 v. 37.6%). However, the acrosomal membrane was damaged in epididymal sperm group at 3-h incubation period (15.2 v. 36.0%). The addition of seminal plasma [0, 10, 15, 20% (v/v)] in Modena solution protected the acrosomal injury (3 h; 35.2, 19.5, 15.6, 14.6%) and maintained high rate of motility (6 h; 38.8, 48.8, 62.5, 60.0%) in a dose-dependent manner. Furthermore, the addition of seminal plasma suppressed the expression of the 15 kDa phosphoprotein (early capacitation status), and the maximum effect was detected at 15% (v/v) seminal plasma. When the frozen–thawed epididymal spermatozoa with 15% (v/v) seminal plasma were artificially inseminated to swine (n = 15), the conception rate and the mean number of litter size were increased as compared with control (93 v. 43%, 10.0 v. 5.0). From these results, we concluded that the addition of seminal plasma to thawing solution was a beneficial method for artificial insemination using frozen–thawed epididymal spermatozoa in the pig. This work was supported by the Programme for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry, and JST-Grant (No. 12-068 and No. 12-104).

228 Effect of seminal plasma on the interval to application of equine chorionic gonadotrophin for the recovery of cumulus-oocyte complexes in alpacas (Vicugna pacos)

2020 ◽  
Vol 32 (2) ◽  
pp. 242
Author(s):  
W. F. Huanca ◽  
J. M. Palomino ◽  
J. C. Villanueva ◽  
J. Malaga ◽  
W. Huanca
Keyword(s):  
Seminal Plasma ◽  
Chorionic Gonadotrophin ◽  
Limited Information ◽  
Dominant Follicle ◽  
Follicular Wave ◽  
Β Protein ◽  
Vicugna Pacos ◽  
Group Data ◽  
Group 2

Alpacas as other camelids are induced ovulators and require an external stimulus-mounting by the male-for ovulation. NGF-β, protein present in the seminal plasma (SP) is reported as being responsible for stimulating ovulation. However, limited information exists on the effect of ovulation inducers in the new follicular wave emergence with application on protocols of superstimulation. The aim of the study was to determine the effect of two ovulation induced techniques and two different times on the interval to application of equine chorionic gonadotrophin (eCG) for the ovarian superstimulation for the recovery of COCs by ovum pick- up (OPU). Alpacas were evaluated by transrectal ultrasonography with an Aloka SSD 500 ultrasound and transducer (7.5MHz) to determine the presence of a dominant follicle=7mm. A 2×2 experimental design was used with adult female alpacas, 6 to 8 years old, which were assigned to one of the treatments when a dominant follicle=7mm was present. Alpacas were assigned randomly 1 of 2 groups (Day 0) in which follicular ovulation was induced by seminal plasma 1mL IM (SP, n=13; group 1) with application of 650IU of eCG at 36h (n=6) or 48h (n=7) post-application of SP, or by GnRH with 0.008mg of Buserelin IM (GnRH, n=10; group 2) with application of 650IU eCG at 36h (n=5) or 48h (n=5) post-application of GnRH. On Day 7, COCs were counted and the OPU procedure was performed in every group. Data were analysed using ANOVA. Results are presented in Table 1. In conclusion, both inducers of ovulation and both times of application of eCG were effective for producing follicles of 7mm and COCs acceptable for recovery in Alpacas. Table 1.Number of follicles=7mm (top) and number of COCs recovered (mean±s.e.m.) Outcome and group 36 h 48 h Total Follicles GnRH 6.8±1.93 13.4±4.34 10.10±2.50a SP 5.5±2.23 5.29±1.15 5.38±1.25b Total 6.09±1.44 8.67±2.18 7.43±1.33 COCs GnRH 5±0.97 3±1.26 4.33±0.74a SP 1±0.41 0.33±0.22 0.67±0.25b Total 3.29±0.67x 1.4±0.6y 2.5±0.53 a,bValues within columns with different letters differ significantly (P=0.05). x,yValues within columns with different letters differ significantly (P=0.05). This research funded by CIENCIACTIVA-CONCYTEC as part of the project title “Role of seminal plasma in reproductive physiology and application of biotechnologies in camelids” (149-2017).

100 Use of Seminal Plasma, Human Chorionic Gonadotropin, and Follicular Ablation on the Interval to Follicular Wave Emergency and Dominant Follicle in Alpacas (Vicugna pacos)

2018 ◽  
Vol 30 (1) ◽  
pp. 189
Author(s):  
W. F. Huanca ◽  
F. Y. Hilari ◽  
J. C. Villanueva ◽  
M. Uchuari ◽  
W. Huanca
Keyword(s):  
Chorionic Gonadotropin ◽  
Seminal Plasma ◽  
Post Treatment ◽  
Dominant Follicle ◽  
Ultrasound Guided ◽  
External Stimulation ◽  
Follicular Wave ◽  
Vicugna Pacos ◽  
Treatment Data ◽  
Treatment Table

Alpacas, as other camelids, are inducer ovulators and FIO/NGF-β, a protein present in the seminal plasma (SP) is reported as the responsible of the ovulation (Kershaw-Young et al. 2012 Reprod. Fertil. Dev. 24, 1093-1097, 10.1071/RD12039). However, limited and controversial information exists regarding characteristics of follicular wave in alpacas post-induction of ovulation with SP or other stimulus. The experiment was designed to determine the effect of 3 external stimulations on the interval to follicular wave emergence and the interval to dominant follicle. Adult female alpacas between 5 and 6 years old were assigned to 1 of 3 treatments: (1) SP (n = 6): 1 mL of SP IM; (2) hCG (n = 5): 1000 IU of hCG (Pregnyl, Organon-Holland, Amsterdam, the Netherlands), via IM; or (3) follicular ablation (FA, n = 6): animals were induced by ultrasound-guided ablation of the dominant follicle ≥7 mm. Alpacas from treatments 1 and 2 were examined by ultrasonography (Aloka SSD 500, transducer 7.5 MHz; Aloka, Tokyo, Japan) at 1- to 2-h intervals between 22 and 30 h after treatment or until ovulation occurred, whichever occurred first. All animals were evaluated by ultrasonography every day from Day 2 to Day 7 post-treatment and after that on Days 9, 12, and 15 post-treatment. Data from one alpaca (FA group) was excluded because of problems in the timing of ablation. Therefore, the total number of alpaca used was 16 (SP = 6, hCG = 5, and FA = 5). Results of the effect in external stimulation were analysed using ANOVA. In conclusion, interval to the emergence of a new follicular wave on the detection of follicles ≥3 mm and interval to dominant follicle ≥7 mm differed for FA compared with hCG but not compared with SP treatment. Table 1.Follicular wave emergence (mean ± SEM) under 3 external stimulations: seminal plasma (SP), hCG, or follicular ablation (FA)

70 Effect of Ovulation Stimulus on Biochemical Composition of the Oviduct of Alpaca (Vicugna pacos)

2018 ◽  
Vol 30 (1) ◽  
pp. 173
Author(s):  
F. Y. Hilari ◽  
J. C. Villanueva ◽  
W. F. Huanca ◽  
B. Lira ◽  
W. Huanca
Keyword(s):  
Biochemical Composition ◽  
Ovulation Induction ◽  
Control Group ◽  
Gnrh Analogue ◽  
Adult Females ◽  
Vicugna Pacos ◽  
Buserelin Acetate ◽  
Biochemical Analyzer ◽  
Oviduct Fluid

Experiments were designed to determine the effect of ovulation stimulus on the biochemical composition of oviduct fluid in alpacas (Vicugna pacos). Twenty-four adult females were used to determine profiles of protein, albumin, glucose, cholesterol, and triglycerides in oviduct fluid. The animals were stimulated with 1 mL of seminal plasma (SP), 0.042 mg of a gonadotropin-releasing hormone (GnRH) analogue (buserelin acetate; Conceptal, Intervet, Holland), or PBS as control group. The oviduct was removed inmediately after slaughter on Day 2 or 4 after ovulation induction and washed with 3 mL of PBS solution. Determination of the biochemical composition was carried out with specific kits from the FAR Diagnostics Italia laboratory using a semiautomatic biochemical analyzer of the Sinowa China brand, which is based on spectrophotometric evaluation. The results are presented in Table 1. Our results suggest that the method of induction of ovulation affected the biochemical composition of oviduct fluid, including concentrations of albumin, cholesterol, total protein, triglyceride, and glucose. Table 1.Biochemical composition of oviduct fluid (mean ± SEM).

THE ACCUMULATIVE EFFECT OF FINNSHEEP BREEDING IN CROSSBREEDING SCHEMES: REPRODUCTIVE PERFORMANCE

1988 ◽  
Vol 68 (1) ◽  
pp. 69-81 ◽  
Author(s):  
MOHAMED H. FAHMY ◽  
JACQUES J. DUFOUR
Keyword(s):  
Body Weight ◽  
Mortality Rate ◽  
Key Words ◽  
Litter Size ◽  
Reproductive Performance ◽  
Conception Rate ◽  
Ovulation Rate ◽  
Genetic Groups ◽  
Linear Regressions ◽  
Size At Birth

Reproductive performance and body weight were studied on 361 ewes, representing Finnsheep (F), DLS (a population of 1/2 Dorset, 1/4 Leicester, 1/4 Suffolk) and seven combinations ranging from 1/8 to 7/8 Finnsheep breeding. Conception rate in yearlings was 61.5% for DLS compared to 89.0% for F with the crosses being intermediate. Conception rate in older ewes was similar in the different genetic groups (avg. 94%). Ovulation rate and litter size at birth of DLS ewes were 1.72 and 1.44 lambs, which was less than half those of F ewes (3.51 and 2.86 lambs, respectively). Both traits increased progressively with an increase in F breeding in crosses and with advances in age. DLS ewes weaned 1.22 lambs compared to 2.03 lambs for F ewes and 1.84 lambs for 4/8 F ewes. The heaviest litters at weaning (31.7 kg) were raised by 4/8 F ewes, followed by 7/8 F (30.8 kg) while those raised by DLS ewes weighed 23.0 kg and F ewes 29.1 kg. Percentage of ova lost per ewe mated averaged 24% and ranged between 18% (DLS and 1/8 F) and 29% (6/8 F). About 3.6% of lambs were born dead and a further 13.8% died before weaning. Preweaning mortality rate was highest in F (22.9%) and lowest in 3/8 F (9.4%). Average kilograms of lambs weaned per ewe exposed was highest in 4/8 F (27.6 kg) followed by F (26.0 kg), whereas that of DLS was the lowest at 18.1 kg. The 4/8 F cross showed 25% heterosis in kg of lambs weaned per ewe exposed and 52.5% increase over DLS. Significant positive linear regressions were calculated for ovulation rate, litter size and preweaning mortality rate on proportion of Finnsheep breeding in crosses. The relation was quadratic for percent ova lost and lamb mortality at weaning. Yearling DLS females weighted 36 kg compared to 44 kg for F yearlings. However, at 5 yr of age DLS ewes weighed 62 kg, 5 kg heavier than F ewes. The heaviest ewes at all ages were the 4/8 F (45 kg at 1 yr, 65 kg at 5 yr). Key words: Reproduction, DLS sheep, Finnsheep, crossbreeding, heterosis, repeatabilities

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