Embryo recovery rate in Santa Inês ewes subjected to successive superovulatory treatments with pFSH

The objective of this study was to evaluate the correlation between the number of corpora lutea (CL), as determined by B-mode ultrasonography, and the total number of structures recovery by non-surgical embryo recovery (NSER) from superovulated ewes. Somali (n=18), Santa Inês (n=18), and Morada Nova (n=16) ewes received an intravaginal device with progesterone (0.33g, controlled internal drug release device, CIDR, Zoetis), which was maintained for 9 days. The superovulatory protocol started 60h before device removal, with 6 decreasing doses (25, 25, 15, 15, 10, and 10%) of p-FSH (IM 133mg, Folltropin V, Vetoquinol) injected every 12h. D-Cloprostenol (IM 37.5μg, Prolise, Agener Union) was injected 12h before and at the moment of device removal. Ewes in oestrus were mated by fertile male rams three times after the onset of oestrus with a 12-h interval. Flunixin meglumine (24.9 mg; Banamine, MSD Animal Health) was injected IM on Days 12, 13, and 15. One day before NSER, B-mode ultrasound evaluations were performed using portable equipment (Z5 Vet, Mindray) with a stiffened multifrequency linear probe to evaluate the number of structures present in the ovaries. Embryo recovery was performed 7 days after progesterone CIDR removal (Day 16). The number of CL and the number of recovered structures were determined and their association evaluated using the Pearson correlation test (P<0.05). Only 17 Somali, 16 Santa Inês, and 15 Morada Nova ewes were submitted to NSER; the other 4 ewes could not be flushed because of low cervical dilation or cervical puncture. After B-mode ultrasound evaluation, 168, 217, and 131 CL were observed, with an average of 9.9 (range: 1 to 12), 13.5 (range: 2 to 47), and 8.7 (range: 3 to 18) CL per donor for Somali, Santa Inês, and Morada Nova ewes, respectively. The recovery rates (i.e. structures recovered by the number of CL counted) were 60.1% (101/168), 96.3% (209/217), and 103.5% (135/131) for the 3 breeds, respectively. There was a positive correlation (r=0.69; P<0.01) between the number of recovered structures and number of CL. Those findings confirm results from other studies, in which the recovery rate varied from 35 to 91%. In some cases, recovery rate can be overestimated because of the difficulty in quantifying CL. In conclusion, B-mode ultrasonography can be used to estimate superovulatory response in ewes and thus as a criterion to decide whether a donor will undergo embryo flushing. Financial support for this study was provided by Embrapa (02.13.06.026.00.04) and Fapemig (CVZ-PPM 00201-17).

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Successive in vivo embryo production in Santa Inês sheep

Animal Production Science ◽

10.1071/an18740 ◽

2020 ◽

Vol 60 (4) ◽

pp. 497 ◽

Cited By ~ 4

Author(s):

Pedro Henrique Nicolau Pinto ◽

Mario Felipe Alvarez Balaro ◽

Helena Fabiana Reis de Almeida Saraiva ◽

Viviane Lopes Brair ◽

Vivian Angélico Pereira Alfradique ◽

...

Keyword(s):

Recovery Rate ◽

Doppler Ultrasonography ◽

Corpora Lutea ◽

Colour Doppler ◽

Embryo Production ◽

Previous Response ◽

Embryo Recovery ◽

Colour Doppler Ultrasonography ◽

Santa Inês

Context In vivo embryo production, also called multiple ovulation and embryo transfer, can accelerate genetic gain, and thus improve animal production. However, there are issues limiting a wider use of this biotechnology in sheep livestock. Aims This study aimed to determine (1) whether a previous response to superovulation (SOV) can be used as a criterion to select ewes for in vivo embryo production, (2) whether the intensity of the SOV response (number of corpora lutea, CL) can affect the embryo recovery rate, and (3) whether the number of CL quantified by colour Doppler ultrasonography can be used to calculate the recovery rate. Methods Twenty-five Santa Inês ewes underwent SOV three times (SOV1, SOV2 and SOV3), with 200 mg FSH and natural mating. The number of CL after each SOV was determined by laparoscopy and by colour Doppler ultrasonography. Key results The number of CL significantly decreased (P < 0.05) after SOV1 (7.5 ± 4.8) to 3.0 ± 5.0 at SOV 2 and 2.2 ± 3.5 at SOV3. Strong correlations were observed between SOV2 and SOV3 in terms of numbers of CL (r = 0.86, r2 = 0.74; P < 0.0001) and viable embryos (r = 0.79, r2 = 0.63; P < 00001). However, no correlations were observed between SOV1 and SOV2 or between SOV1 and SOV3. Recovery rate did not differ with the intensity of the SOV response (≤6, 7–10, >10 CL) or between the methods used to quantify CL. Conclusions Ewes did not show the same pattern of response when submitted to successive FSH-based SOV. The intensity of the SOV response did not affect the recovery rate, and the number of CL estimated by colour Doppler ultrasonography can be used to calculate the recovery rate. Implications Selecting sheep embryo donors by a previous SOV response is not always feasible. The recovery rate is homogeneous and it is not affected by the intensity of the SOV response. A nonsurgical technique can be used to assess the recovery rate, improving animal welfare in MOET programs.

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Effect of season and frequency of embryo collections on superovulatory response and embryo recovery in Santa Inês hair sheep

Small Ruminant Research ◽

10.1016/j.smallrumres.2021.106441 ◽

2021 ◽

pp. 106441

Author(s):

Monique de Albuquerque Lagares ◽

Fabiana Cristina Varago ◽

Valéria Spyridion Moustacas ◽

Valentim Arabicano Gheller ◽

Rafael Romero Nicolino ◽

...

Keyword(s):

Hair Sheep ◽

Embryo Recovery ◽

Santa Inês

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Leptospirosis and embryo recovery rate in mares: Table 1

Veterinary Record ◽

10.1136/vr.e296 ◽

2012 ◽

Vol 170 (2) ◽

pp. 60-60 ◽

Cited By ~ 5

Author(s):

A. Pinna ◽

G. Martins ◽

W. Lilenbaum

Keyword(s):

Recovery Rate ◽

Embryo Recovery

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Embryo Recovery Rate in a Mare Affected by Cushing's Syndrome

Veterinary Research Communications ◽

10.1023/b:verc.0000014231.45688.ac ◽

2003 ◽

Vol 27 ◽

pp. 619-621 ◽

Cited By ~ 1

Author(s):

D. Panzani ◽

I. Vannozzi ◽

M. Sgorbini ◽

M. Corazza ◽

A. Rota ◽

...

Keyword(s):

Cushing’S Syndrome ◽

Recovery Rate ◽

Cushing's Syndrome ◽

Embryo Recovery

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171IMPROVEMENT IN EMBRYO RECOVERY USING UTERINE DOUBLE FLUSHING

Reproduction Fertility and Development ◽

10.1071/rdv16n1ab171 ◽

2004 ◽

Vol 16 (2) ◽

pp. 207 ◽

Cited By ~ 1

Author(s):

A. Castro Neto ◽

B.V. Sanches ◽

S.H.V. Perri ◽

M.M. Seneda ◽

J.F. Garcia

Keyword(s):

Recovery Rate ◽

Foley Catheter ◽

Test Group ◽

Control Group ◽

Recovery Rates ◽

Mean Values ◽

Embryo Recovery ◽

Group A ◽

Uterine Flushing ◽

The Mean

The aim of the present study was to evaluate the effects of uterine double flushing on embryo recovery rates (total structures and viable embryos), after a resting period with the uterus filled with PBS. There were 210 embryo recovery procedures conducted using the uterine double flushing method, and the results were compaired with 432 conventional single flushing procedures. All procedures were conducted with Limousin (n=403) and Guzerá (n=239) cows, following the same superovulation protocol. Cyclic donors received a progestagen implant (CIDR) and 1mL of oestradiol benzoate (Estrogin®) at Day 0. Between Day 5 and Day 9 animals received doses ranging from 200 to 300UI (zebu cows) and 300 to 500UI (taurine cows) of FSH (Pluset® – Serono) in decreasing doses. Between Day 6 and Day 8 PGF2a (Ciosin®) was administered, followed by withdrawal of the progestagen after 24h. Artificial insemination was performed between 14 and 26h after the beginning of treatment. For the double flushing procedure, after the first uterine flushing of both horns with 1L DPBS (Nutricell), a Foley catheter was positioned in the uterine body in order to fill the uterus with the same solution (80 to 150mL). After this procedure the catheter was closed with a disposable 5-mL syringe gasket, and the animals were allowed to rest in the surroundings of the work place for 30min. After this period, a second flush was performed in order to recover the remaning liquid used during manipulation of the uterus. Animals from the control group (group A) were subjected to a single uterine flushing procedure. From 210 double flushing procedures (group B – test), 1409 viable embryos were produced, classified as grades I, II, III and IV (IETS), (average of 6.7 embryos per procedure), whereas, in the 432 single flushing procedure (group A – control), 1993 embryos were produced (average of 4.6). Statistical analysis showed the increase of viable embryo recovery rate. When consecutive double flushing was performed, the average of recovered embryos increased from 8.3 to 12.7 (P<0.05) in Limousin cows, and from 7.9 to 11.5 (P<0.05) in Guzerá cows. Comparing recovery after single flushing with that after double flushing, the mean number of viable embryos increased from 4.7 to 6.9 (P<0.05) in Limousin cows and from 4.5 to 6.4 (P<0.05) in Guzerá cows. In order to assure the nonexistence of negligence effects or operator influence on results, the mean values of total embryo recovery rate after single flushing (control group) was compared to the mean values of the same rate after the first uterine flushing on test group. Results indicated no difference between recovery rates. The present work showed the viability of using the uterine double flushing procedure for improvement of embryo recovery rates in cattle.

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Embryo recovery from the oviduct in superovulated ewes: a method to improve MOET systems

Czech Journal of Animal Science ◽

10.17221/374-cjas ◽

2008 ◽

Vol 53 (No. 4) ◽

pp. 145-151

Author(s):

J.P. Ramon-Ugalde ◽

J. Folch ◽

M.J. Cocero ◽

R.E. Piña-Aguilar ◽

J.L. Alabart

Keyword(s):

Recovery Rate ◽

Uterine Horn ◽

Conception Rate ◽

The Other ◽

Embryo Recovery ◽

Intramuscular Injections ◽

Other Hand

The efficiency of embryo recovery in a superovulatory treatment was studied by perfusing the oviduct or the uterine horn in 3.5 and 7 days after sponge withdrawal, respectively. Eighty-four and seventy ewes of Aragonesa breed were used as embryo donors and receptors, respectively. The donors were distributed in 3 replications. The oestrus was synchronized with the insertion of FGA intravaginal sponges (30 mg) for 13 days. Six intramuscular injections of 18 mg pFSH were applied in decreasing doses at 12 hour intervals starting 48 hours after sponge withdrawal. The recovery rate (RR) (83.2 vs. 75.8%), the viability rate (VR) (73.5 vs. 47.2%) and the number of viable embryos (VE) per donor ewe (5.9 ± 0.79 vs. 3.0 ± 0.37) were higher (<I>P</I> < 0.01) in ewes whose perfusion was done directly in oviducts. The interval from sponge withdrawal to oestrus had no influence on RR in any groups; however it had an effect on the VR in 3.5 and 7 days old embryos. Embryos from ewes showing the oestrus within 20 hours after sponge withdrawal had a higher viability. On the other hand, using morphologic criteria, 26.5% and 52.7% of 3.5 and 7 days old embryos, respectively, were discarded before transfer. The percentage of ewes in each group that became pregnant was similar in both groups. We conclude that in superovulated ewes the embryo recovery rate, viability and number of viable embryos are higher in embryos recovered from the oviduct compared to those recovered from the uterus, without decreasing the conception rate and the viability rate is higher in ewes that show the oestrus 20 hours before sponge withdrawal.

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108 COMPARATIVE EFFICIENCY OF GONADOTROPIN-RELEASING HORMONE AND LUTEINIZING HORMONE IN THE INDUCTION OF OVULATION IN SUPEROVULATED ALPACAS

Reproduction Fertility and Development ◽

10.1071/rdv26n1ab108 ◽

2014 ◽

Vol 26 (1) ◽

pp. 168

Author(s):

H. W. Vivanco-Mackie ◽

M. D. Ponce Salazar ◽

M. M. Gonzales ◽

M. A. Tapia

Keyword(s):

Gnrh Agonist ◽

Recovery Rate ◽

Pearson Correlation ◽

Animal Health ◽

Gonadotropin Releasing Hormone ◽

Corpora Lutea ◽

Peruvian Andes ◽

Releasing Hormone ◽

Induction Of Ovulation ◽

Embryo Recovery

Alpacas are induced ovulators, responding to copulation and (or) exogenous application of ovulation-inducing hormones. The objective of this study was to determine the efficiency of the injection of a gonadotropin-releasing hormone (GnRH) agonist versus LH in the induction of ovulation and the presence and size of non-ovulated follicles at the time of embryo collection and its relationship to the yield of transferable embryos in superovulated alpacas. Twenty-one adult (3 to 7 years old) female alpacas under extensive grazing at 4300 m elevation in the Peruvian Andes that had been synchronized and treated for superovulation were induced to ovulate 6 days after the application of the superovulatory hormone (1000 IU of eCG, Folligon®, Intervet International BV, Boxmeer, the Netherlands) by mating with fertile males and injection immediately after copulation of either an IM dose of 0.0084 mg of buserelin acetate (Buserelina Zoovet®, Laboratorio Zoovet, Santa Fe, Argentina) to 10 alpacas (T1) or an IM dose of 5-mg Armour standard of LH (Lutropin®, Bioniche Animal Health, Belleville, ON, Canada) to 11 alpacas (T2). All alpacas had a second mating 12 h after the first mating and were subjected to ovarian inspection by ultrasonography and previous embryo collection by nonsurgical transcervical embryo flushing 6.5 days after the first mating. On average, the embryo recovery rate for T1 was 34.6% and there were 7.8 ± 3.99 corpora lutea (CL), 2.7 ± 4.08 collected embryos, and 3.6 ± 2.95 total, 0.5 ± 0.85 small (<6 mm), 1.8 ± 1.99 medium (≥6 but ≤14 mm), and 1.3 ± 2.11 large (≥15 mm) non-ovulated follicles. For T2, the embryo recovery rate was 59.4% and there were 6.73 ± 1.49 CL, 4.0 ± 2.57 collected embryos, and 0.64 ± 0.81 total, 0.0 ± 0.0 small, 0.36 ± 0.67 medium, and 0.27 ± 0.47 large non-ovulated follicles. The differences between treatments were nonsignificant (P > 0.05) for all the parameters; however, there was a clear tendency for a better recovery rate, more embryos collected, and lower number of non-ovulated follicles in T2. The Pearson correlation coefficient between the presence of large follicles in the ovaries at the time of embryo collection and the total number of embryos collected was negative (r = –0.26) and highly significant (P ≤ 0.001). The use of LH for ovulation induction of superovulated alpacas seems to be more advisable than the use of GnRH agonist; further comparisons with larger number of observations per treatment are recommended. This study was financed by the Peruvian Fund for Innovation, Science and Technology (FINCYT).

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157 EFFECT OF DENSITY GRADIENT SELECTION ON EMBRYO RECOVERY RATES OF FRESH AND COOLED STALLION SEMEN

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab157 ◽

2013 ◽

Vol 25 (1) ◽

pp. 227

Author(s):

F. Papa ◽

M. Carmo ◽

P. Papa ◽

J. Dell'Aqua ◽

M. Alvarenga

Keyword(s):

Plasma Membrane ◽

Density Gradient ◽

Recovery Rate ◽

Ovulation Induction ◽

Membrane Integrity ◽

Recovery Rates ◽

Plasma Membrane Integrity ◽

Embryo Recovery ◽

Fresh Semen ◽

Sperm Recovery

The aim of this study was to improve the spermatic kinetic parameters from stallions with poor quality of fresh and chilled semen by the use of density gradient Equipure™ (Nidacon, Mölndal, Sweden). Semen from 5 Quarter Horse stallions aged 8 and 16 years with history of low embryo recovery rates were used. The kinetics sperm evaluation was performed by computerized semen analysis (CASA) and plasma membrane integrity with fluorescent probes. The average motility parameters for fresh semen before selection were total motility (MT) 60%, progressive motility (PM) 30%, and plasma membrane integrity (IMP) 60% and for cooled semen (24 h at 5°C) were: MT 50%, MP 18%, and 50% IMP. For the group of fresh and cooled semen with no density gradient selection (NS), mares were inseminated with 1 billion viable sperm diluted in skim milk extender in the uterine body, 24 h after ovulation induction with 1 mg of deslorelin. For EquipureTM selection group (SE) semen was concentrated through Spermfilter membrane™ and resuspended in 5 mL of BotuSemenTM (BotupharmaTM, Brazil). In a 15 mL conic tube 5 mL of EquipureTM was added and another part containing 5 mL of the resuspended sperm was slowly added over the EquipureTM column. The 15 mL conic tube was centrifuged at 400g for 20 min. After centrifugation, the sperm pellet was carefully aspirated and to the pellet was resuspended in 4 mL of BotucrioTM. The sperm recovery rate with EquipureTM was 40%. Deep uterine AI was performed 24 h after ovulation induction with 1 mg of deslorelin. Data were analyzed by ANOVA followed by Tukey’s test (P < 0.05). The analysis of semen after EquipureTM selection resulted in average: 75, 35, 65, and 40%, respectively, for MT, MP, and IMP index and sperm recovery. The embryo recovery rate from the 5 stallions showed the following results: Stallion 1 (fresh semen): 12 mares (NS)/4 embryos (33%) and for group SE 22 mares/16 embryos, (72%; P ≤ 0.05). Stallion 2 (fresh semen): 9 mares (NS)/4 embryos, (44%) and for group SE, 12 mares/8 embryos (66%; P > 0.1). Stallion 3 (fresh semen): 4 mares (NS)/1 embryo (25%) and 7 mares for SE/4 embryos (57%; P > 0.1). Stallion 4 (chilled semen): 4 mares (NS)/0 embryo 0% and for SE group 8 mares/6 embryos 75%; P ≤ 0.001). Stallion 5 (chilled semen): 10 mares (NS)/2 embryos (20%) and for SE 6 mares/3 embryos (50%; P > 0.1). For the overall results, 39 inseminations were performed on the no selected group and 11 embryos recovered (28%) for the selected group 55 inseminations and 37 embryos recovered (67%; P < 0.01). The results clearly showed that selection by sperm density gradient EquipureTM was a very effective technique that allowed an improvement on semen quality and fertility. The authors acknowledge support from FAPESP and Botupharma.

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407 SUPERSTIMULATORY RESPONSE WITH DECREASING DOSES OF FOLLICLE-STIMULATING HORMONE v. A SINGLE DOSE OF FOLLICLE-STIMULATING HORMONE - EQUINE CHORIONIC GONADOTROPIN DURING BREEDING AND NONBREEDING SEASON IN SHEEP

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab407 ◽

2010 ◽

Vol 22 (1) ◽

pp. 360

Author(s):

M. I. Cueto ◽

F. Pereyra-Bonnet ◽

P. Silvestre ◽

A. E. Gibbons

Keyword(s):

Single Dose ◽

Breeding Season ◽

Recovery Rate ◽

Follicle Stimulating Hormone ◽

Ovulation Rate ◽

Corpora Lutea ◽

Nonbreeding Season ◽

Embryo Recovery ◽

The One ◽

Stimulating Hormone

The aim of the study was to assess possible variations in superovulatory yields due to different FSH treatments at 2 times of the year. Superovulation and embryo recovery were performed during the breeding (n = 63) andnonbreeding (n = 46) seasons in Merino ewes located at 41°S latitude. Animals were kept under the same conditions, housed outdoors in a sheltered and covered pen, and were fed a liveweight maintenance ration. All animals received 60-mg medroxyprogesterone acetate intravaginal sponges (Progespon®, Syntex, Buenos Aires, Argentina) for 14 days. Ewes were then randomly assigned to 2 different superovulatory treatments: classic (n = 74) and one shot (n = 35) in both seasons. Classic superovulatory treatment consisted of 7 decreasing doses (2 × 48 mg, 2 × 24 mg, 2 × 20 mg, and1 × 16 mg NIH-FSH-P1)ofFSH (Folltropin®-V, Bioniche, Belleville, Ontario, Canada), administered twice daily from 48 h before to 24 h after pessary removal. A dose of eCG (300 IU; Novormon®, Syntex) was administered at progestagen removal. One shot superovulatory treatment consisted of a single dose of FSH (70 mg NIH-FSH-P1) plus 300 UI of eCG injected at pessary withdrawal. Embryo donors were inseminated by laparoscopy with frozen-thawed semen (100 × 106 spz) 12 h after the onset of estrus. Surgical embryo recovery was done on Day 7 after sponge withdrawal and embryos were graded for quality according to morphology (Grade 1 = excellent or good; Grade 2 = fair; Grade 3 = poor; and Grade 4 = dead or degenerated; IETS 1998). A 2 × 2 factorial ANOVA was used to test the main effects (season and superovulatory treatment) and interactions. There were no significant differences in the proportion of responding ewes (>3 corpora lutea), ovulation rate, and recovered Grades 1 to 2 embryos between the breeding and nonbreeding season (Table 1; P > 0.05). However, number of recovered ova/embryos and ova/embryo recovery rate were higher during the breeding season compared with the nonbreeding season, whereas the percentage of nonfertilized ova was lower in the breeding season than in the nonbreeding season (P < 0.05). Analysis of data comparing superovulatory treatments showed that the proportion of responding ewes, ovulation rate, recovered embryos, and recovered Grades 1 to 2 embryos were lower for the one shot treatment than for the classic treatment (P < 0.05). Embryo recovery rate and nonfertilization rate did not differ between treatments (P > 0.05). It was concluded that there was an increase in the number of total recovered ova/embryos during the breeding season compared with the nonbreeding season, although the number of recovered good-quality embryos was not affected. The use of multiple FSH injections produced a higher number of total recovered and viable embryos in Merino sheep than the one shot superovulatory treatment. Table 1.Embryo yields in ewes submitted to superovulation

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