Well-of-the-well (WOW) versus polyester mesh (PM): a comparison of single-embryo culture systems in bovines

Background: Mexico is innovating in the livestock industry through in vitro generation of bovine embryos with technologies such as well-of-the-well (WOW) and polyester mesh (PM) single-embryo culture systems. These techniques allow to maintain embryos in separate areas of a shared culture medium. Objective: To compare the quantity and quality of bovine embryos produced in WOW and PM culture systems versus the conventional (CG) culture system. Methods: In total, 345 embryos fertilized in vitro were evaluated for blastocyst yield in the three culture systems. To count blastocyst cell numbers, 69 embryos in each system were differentially stained for trophectoderm (TE), inner cell mass (ICM), and apoptotic cells. A qPCR gene expression analysis was performed for embryos in all three systems. Results: The WOW, PM and CG systems developed similar amount of blastocysts (41, 35 and 36%, respectively; p>0.05). Blastocysts in all three systems showed adequate amounts of ICM and apoptotic cells. Blastocysts in the PM system showed a greater number of TE cells [63.7 versus 58.6% in the CG system (p0.05). The ATP5B expression was higher in WOW than in PM (p0.05). The TJP3 expression was higher in PM than in WOW and CG (p<0.05). Expression of ID2 and CLDN4 was higher in WOW than in PM and CG (p<0.05). The biplot graphic from Principal Component Analysis (PCA) revealed that CG was located near degenerated embryos, whereas PM was located near arrested embryos, larger ICM and TE, and TJP3 expression. The WOW was located toward blastocysts, morulae, and expression of CLDN4, ID2 and GNAS. Conclusion: Compared with CG, both the PM and WOW systems are good options for culturing single embryos in the bovine model. Moreover, the PCA results suggest that embryos developed in the WOW system have greater capacity for generating blastocysts with increased ability to form TE and ICM layers, which might improve implantation.

Antecedentes: México está innovando en la industria ganadera a través de la generación in vitro de embriones bovinos con tecnologías de cultivo individual como lo son Pozo dentro de Pozo (WOW) y Malla de Poliéster (PM). Estos mantienen los embriones en áreas separadas mientras comparten un mismo medio de cultivo celular. Objetivo: Comparar la cantidad y calidad de embriones bovinos producidos en los sistemas WOW y PM contra el sistema de cultivo convencional en grupo (CG). Métodos: En total se evaluaron 345 embriones fertilizados in vitro para determinar la producción de blastocistos generados en los tres sistemas. Para contar el número de células por blastocisto, 69 embriones en cada sistema se tiñeron diferencialmente para trofectodermo (TE), masa celular interna (ICM) y células apoptóticas. Se realizó un análisis de expresión génica por qPCR de los embriones obtenidos en los tres sistemas. Resultados: Los sistemas WOW, PM y CG desarrollaron similares cantidades de blastocistos (41, 35 y 36%, respectivamente; p>0,05). Los blastocistos en los tres sistemas mostraron cantidades adecuadas de ICM y células apoptóticas. Los blastocistos en el sistema PM mostraron un mayor número de células TE [63,7% versus 58,6% en el sistema CG (p0,05). La expresión de ATP5B fue mayor en WOW que en PM (p<0,05), pero similar a CG (p<0,05). La expresión de TJP3 fue mayor en PM que en WOW y CG (p<0,05). La expresión de ID2 y CLDN4 fue mayor en WOW que en PM y CG (p<0,05). El gráfico de biplot del análisis de componentes principales reveló que CG se encontró cerca de embriones degenerados, mientras que PM se encontró cerca de embriones en arresto, ICM, TE, y TJP3. El WOW se localizó hacia blastocistos, mórulas y la expresión de CLDN4, ID2 y GNAS. Conclusión: En el modelo bovino los sistemas PM y WOW son buenas opciones para cultivar embriones individuales, ya que se obtienen resultados muy similares a los obtenidos con el sistema CG. Además, los resultados de PCA sugieren que los embriones individuales desarrollados en el sistema WOW generan blastocistos con mayor capacidad de formar TE e ICM, lo que podría mejorar su éxito de implantación.

Antecedentes: O México está inovando na indústria pecuária por meio da geração in vitro de embriões bovinos com tecnologias de cultura de embriões individuais, bem como em poço (WOW) e malha de poliéster (PM). Estes mantêm os embriões em áreas separadas, enquanto compartilham o mesmo meio de cultura de células. Objetivo: Comparar a quantidade e a qualidade de embriões bovinos produzidos nos sistemas de cultura WOW e PM com o sistema convencional de cultura em grupo (CG). Métodos: No total, 345 embriões fertilizados in vitro foram avaliados para determinar a produção de blastocistos gerados nos três sistemas. O número de células por blatocisto foi contado, 69 embriões em cada sistema foram diferencialmente corados para trofectoderme (TE), massa celular interna (ICM) e células apoptóticas. Uma análise de expressão gênica qPCR foi realizada para os embriões obtidos nos três sistemas. Resultados: Os sistemas WOW, PM e CG desenvolveram quantidades semelhantes de blastocistos (41, 35 e 36%, respectivamente; p>0,05). Os blastocistos nos três sistemas mostraram quantidades adequadas de ICM e células apoptóticas. Os blastocistos no sistema PM mostraram um número maior de células TE [63,7 versus 58,6% no sistema CG (p0,05). A expressão de ATP5B foi maior no WOW do que no PM (p<0,05), mas semelhante ao GC (p<0,05). A expressão de TJP3 foi maior no PM do que no WOW e CG (p<0,05). A expressão de ID2 e CLDN4 foi maior no WOW do que no PM e CG (p<0,05). O gráfico biplot da análise de componentes principais revelou que CG foi encontrado próximo a embriões degenerados, enquanto PM foi encontrado próximo a embriões presos, ICM, TE e TJP3. WOW foi encontrado para ter blastocistos, mórulas e a expressão de CLDN4, ID2 e GNAS. Conclusão: Em comparação com o CG, os sistemas PM e WOW são boas opções para a cultura de embriões individuais no modelo bovino. Além disso, os resultados da PCA sugerem que embriões individuais desenvolvidos no sistema WOW têm maior capacidade de desenvolver blastocistos com maior capacidade de formar as camadas TE e ICM, o que poderia melhorar seu sucesso de implantação.

Transcervical Versus Laparotomy Embryo Recovery: What Strategy Is Best for Embryo Bank Formation in the Canindé Goat Conservation Program?

By allowing for the creation of embryo banks, reproductive biotechnologies play an essential role in the preservation of endangered goat breeds' genetic diversity. This study focused on comparing both available embryo collection methods [laparotomy (LAP) vs. nonsurgical embryo recovery (NSER)] in Canindé goats to create an embryo bank for later use in a breed conservation program. Twelve females were superovulated and subjected to either the LAP or NSER technique for embryo recovery. The recovery rate was similar (p > 0.05) between NSER (86.8% ± 5.6%) and LAP (92.8% ± 4.0%). Moreover, there were no differences (p > 0.05) in the number of structures recovered, the viable embryos, and the freezable embryos per goat, respectively, for NSER (11.7 ± 1.3, 11.2 ± 1.5, and 10.2 ± 1.1) and LAP (10.3 ± 1.0, 8.7 ± 0.7, and 8.0 ± 0.8). Overall, 132 structures were collected out of 151 ovulations (∼12.6 ± 1.2 corpora lutea per goat). Finally, the procedure duration time was also similar (p > 0.05) for NSER versus LAP, respectively: 32.3 ± 3.3 versus 30.8 ± 3.9 minutes. In conclusion, the NSER method results proved to be similar to the LAP technique in small-sized Canindé goats. It was noticeable, however, that the NSER technique is simpler and provides the possibility for successive procedures with few health risks and sequels for females. This study may hopefully boost in vivo embryo production programs in the Canindé breed, facilitating the formation of embryo banks and so assuring the availability of genetic diversity before any decline becomes irreversible.

Study of the factors affecting embryo yields and quality in superovulated Morada Nova ewes that underwent non-surgical uterine flushing.

The present study compared the outcomes of in vivo embryo production in Morada Nova ewes subjected to either 9-day (G-9SOV , n = 21) or 12-day (G-12SOV , n = 21) progesterone (P4 )-based estruses synchronization protocol coupled with superovulatory treatment with decreasing doses of porcine follicle-stimulating hormone (133 mg of pFSH given over 3 days). Non-surgical embryo recovery (NSER) was performed 6-7 days after the onset of oestrus. Total antral follicle count doubled from the first to the sixth pFSH dose in both groups (p < .05). Oestrus responses did not vary between the two groups of animals (95.2%). Corpora lutea (CL) were detected in 85.0% and 60.0% of ewes that previously manifested oestrus behaviour in G-9SOV and G-12SOV respectively. NSER was successfully completed in 86.2% of ewes that had CL (p > .05). The mean number of CL per ewe/successfully flushed donor ewe was greater (p < .05) in G-12SOV (12.3 ± 1.7/12.1 ± 1.9) than in G-9SOV (7.9 ± 1.4/8.2 ± 1.6). Mean numbers of retrieved blastocysts and viable embryos were greater (p > .05) in G-12SOV (5.8 ± 1.9 and 3.7 ± 1.7) than G-9SOV (3.5 ± 1.1 and 0.8 ± 0.3 respectively). The total follicle count (all follicles ≥2 mm in diameter) at the sixth pFSH dose (at P4 -device removal) was positively correlated (p < .05) with the number of CL (r = .95) and viable embryos (r = .91) in G-12SOV . The ewes with ≥10 Cl (48% of all flushed donors) yielded 80.5% of viable embryos. In summary: (a) Morada Nova ewes from G-12SOV group had better superovulatory responses compared with G-9SOV group; (b) total follicle count at the last pFSH dose was a good predictor of superovulatory responses only in the ewes primed with P4 for 12 days; and (c) animals with ≥10 ovulations are main contributors to viable embryo production in Morada Nova ewes.

Fixed-time artificial insemination protocols on brazilian locally adapted breed gilts on ovulatory response and embryo production.

The aim of this study was to use estrus synchronization protocols to favor fixed-time artificial insemination and consequently fixed-time embryo collection, and increase embryo production using eCG, in gits. In a cross over design, nine Piau breed gilts were subjected to 18 days of oral progesterone; P4 group did not receive any further; GnRH group received 25µg of GnRH 104 hours after the final application of P4; and eCG+GnRH group received 1000IU of eCG 24 hours after the final P4 in addition to GnRH for subsequent embryo collection, that was performed six days after first AI, by laparotomy. Artificial insemination was performed after 12 and 24 hours of estrus in P4 group, and 128 and 144 hours in GnRH and eCG+GnRH groups. The number of CL (8.6±3.9; 8.3±2.1; 26.7±15.0) and anovulatory follicles (4.3±3.7; 3.9±3.9; 17.2±9.5) was higher in the eCG+GnRH gilts (P<0.05). However, the use of 1000 IU of eCG reduced (P<0.05) the number of total structures (5.2±3.6; 5.1±3.1; 1.7±2.7), viable embryos (5.0±3.5; 4.8±3.3; 0.4±0.7), freezable embryos (3.6±3.4; 3.3±3.8; 0.1±0.3) and recovery rate (63.7±38.9; 58.6±24.7; 5.38±9.5). P4 and GnRH protocols were effective in the production and recovery of embryos. However, the use of 1000 IU of eCG, 24 hours after P4, was not effective in promoting the production of embryos, although the animals had superovulated.

Timed artificial insemination in crossbred mares: Reproductive efficiency and costs.

Timed artificial insemination (TAI) has boosted the use of conventional artificial insemination (CAI) by employing hormonal protocols to synchronize oestrus and ovulation. This study aimed to evaluate the efficiency of a hormonal protocol for TAI in mares, based on a combination of progesterone releasing intravaginal device (PRID), prostaglandin (PGF2α ) and human chorionic gonadotropin (hCG); and compare financial costs between CAI and TAI. Twenty-one mares were divided into two groups: CAI group (CAIG; n = 6 mares; 17 oestrous cycles) and TAI group (TAIG; n = 15 mares; 15 oestrous cycles). The CAIG was subjected to CAI, involving follicular dynamics and uterine oedema monitoring with ultrasound examinations (US), and administration of hCG (1,600 IU) when the dominant follicle (DF) diameter's ≥35 mm + uterine oedema + cervix opening. The AI was performed with fresh semen (500 × 106 cells), and embryo was recovered on day 8 (D8) after ovulation. In TAI, mares received 1.9 g PRID on D0. On D10, PRID was removed and 6.71 mg dinoprost tromethamine was administered. Ovulation was induced on D14 (1,600 IU of hCG) regardless of the DF diameter's, and AI was performed with fresh semen (500 × 106 cells). On D30 after AI, pregnancy was confirmed by US. The pregnancy rate was 80.0% in TAIG and 82.3% in CAIG (p > .05). The TAI protocol resulted in 65% reduction in professional transport costs, and 40% reduction in material costs. The TAI was as efficient as CAI, provided reduction in costs and handlings, and is recommended in mares.

Fixed-time artificial insemination protocols on brazilian locally adapted breed gilts on ovulatory response and embryo production

Abstract The aim of this study was to use estrus synchronization protocols to favor fixed-time artificial insemination and consequently fixed-time embryo collection, and increase embryo production using eCG, in gits. In a cross over design, nine Piau breed gilts were subjected to 18 days of oral progesterone; P4 group did not receive any further; GnRH group received 25µg of GnRH 104 hours after the final application of P4; and eCG+GnRH group received 1000IU of eCG 24 hours after the final P4 in addition to GnRH for subsequent embryo collection, that was performed six days after first AI, by laparotomy. Artificial insemination was performed after 12 and 24 hours of estrus in P4 group, and 128 and 144 hours in GnRH and eCG+GnRH groups. The number of CL (8.6±3.9; 8.3±2.1; 26.7±15.0) and anovulatory follicles (4.3±3.7; 3.9±3.9; 17.2±9.5) was higher in the eCG+GnRH gilts (P<0.05). However, the use of 1000 IU of eCG reduced (P<0.05) the number of total structures (5.2±3.6; 5.1±3.1; 1.7±2.7), viable embryos (5.0±3.5; 4.8±3.3; 0.4±0.7), freezable embryos (3.6±3.4; 3.3±3.8; 0.1±0.3) and recovery rate (63.7±38.9; 58.6±24.7; 5.38±9.5). P4 and GnRH protocols were effective in the production and recovery of embryos. However, the use of 1000 IU of eCG, 24 hours after P4, was not effective in promoting the production of embryos, although the animals had superovulated.

Fixed-time artificial insemination protocols on brazilian locally adapted breed gilts on ovulatory response and embryo production

The aim of this study was to use estrus synchronization protocols to favor fixed-time artificial insemination and consequently fixed-time embryo collection, and increase embryo production using eCG, in gits. In a cross over design, nine Piau breed gilts were subjected to 18 days of oral progesterone; P4 group did not receive any further; GnRH group received 25µg of GnRH 104 hours after the final application of P4; and eCG+GnRH group received 1000IU of eCG 24 hours after the final P4 in addition to GnRH for subsequent embryo collection, that was performed six days after first AI, by laparotomy. Artificial insemination was performed after 12 and 24 hours of estrus in P4 group, and 128 and 144 hours in GnRH and eCG+GnRH groups. The number of CL (8.6±3.9; 8.3±2.1; 26.7±15.0) and anovulatory follicles (4.3±3.7; 3.9±3.9; 17.2±9.5) was higher in the eCG+GnRH gilts (P<0.05). However, the use of 1000 IU of eCG reduced (P<0.05) the number of total structures (5.2±3.6; 5.1±3.1; 1.7±2.7), viable embryos (5.0±3.5; 4.8±3.3; 0.4±0.7), freezable embryos (3.6±3.4; 3.3±3.8; 0.1±0.3) and recovery rate (63.7±38.9; 58.6±24.7; 5.38±9.5). P4 and GnRH protocols were effective in the production and recovery of embryos. However, the use of 1000 IU of eCG, 24 hours after P4, was not effective in promoting the production of embryos, although the animals had superovulated.(AU)

Comparison of cattle embryo collection between Ringer's lactate solution and Dulbecco's phosphatebuffered saline

The objective of this study was to determine the efficiency of Ringer's lactate solution (RL) and RL + 1% fetal bovine serum (FBS) and compare them with the efficiency of Dulbecco's phosphate-buffered saline (D-PBS). Twenty-two Wagyu female cattle were subjected to superovulation and were randomly distributed to form three groups: group 1 ­ uterine flushing with RL (n = 8), group 2 ­ uterine flushing with RL + 1% FBS (n = 7), and group 3 ­ uterine flushing with D-PBS (n = 7, control group). Cows received a CIDR® device containing 1.9 g of progesterone at random stages of the estrous cycle (day 0). Progesterone withdrawal occurred on day 8 in the morning. For heifers, 160 mg of porcine follicle-stimulating hormone (FSH-P) was used and for cows, 200 mg. Prostaglandin F2α was also injected on the eighth day of FSH-P administration. On day 9, in the morning, hCG was administered. Females were superovulated and inseminated twice in a fixed time for embryo transfer. On the 16th day, females were subjected to uterine flushing for embryo collection. We collected 76 embryos from 22 females subjected to superovulation, of which 52 were transferable and 24 had degenerated. The total of embryos collected was 23, 16, and 23 for groups 1, 2, and 3, respectively. The embryo recovery rates per group were 13.86±4.23, 15.39±4.61 and 27.16±13.33%, in groups 1, 2, and 3, respectively. The means for the total structures collected per female were 2.88±0.85, 3.00±1.23, and 4.57±1.72 in groups flushed with RL, RL + 1% FBS, and D-PBS, respectively. We conclude that Ringer's lactate solution and Ringer's lactate solution + 1% of FBS and Dulbecco's phosphate-buffered saline showed no significant differences in terms of embryo quality or quantity, suggesting that Ringer's lactate solution is an alternative for collecting embryos in cattle.

Successful transcervical uterine flushing can be performed without or reduced dose of oestradiol benzoate in cervical relaxation protocol in Dorper ewes.

This study assessed the efficiency of cervical relaxation protocol using none, half or full dose (1.0 mg) of oestradiol benzoate in Dorper ewes subjected to non-surgical embryo recovery (NSER). Thirty-six pluriparous ewes received progestogen sponge (60 mg) for 9 days plus eCG administration (300 IU i.m.) 24 hr before sponge removal. Ewes were not mated and were randomly assigned to receive at 16 hr before NSER 37.5 µg d-cloprostenol i.m. and different doses of oestradiol benzoate: 0.0 mg (0EB group; n = 12); 0.5 mg (0.5EB group; n = 12) or 1.0 mg of oestradiol (1.0EB group, n = 12). All ewes received oxytocin (50 IU) i.v. 20 min before NSER, which was performed 8 days after sponge removal. Corpora lutea were counted by transrectal ultrasonography 24 hr before NSER. After procedure, the ewes were kept in natural breeding period to check their post-NSER fertility. NSER was performed in 91.7% (33/36) of the animals with overall fluid recovery efficiency over 97% (p > .05). The cervical transposing with Hegar dilator was longer (p < .05) in 0EB (4.2 ± 0.3 min) compared to 0.5EB (1.7 ± 0.3 min) and 1.0EB group (1.5 ± 0.3 min). The cervical transposing with mandrel/catheter was longer (p < .05) in 0EB (2.4 ± 0.5 min) than 1.0EB group (1.3 ± 0.5 min). Overall duration of uterine flushing was 25.4 min with structure recovery rate of 43.5%, with no difference among groups (p > .05). The post-NSER fertility was higher (p < .05) in 0.0EB (90%) than 0.5EB group (36.4%). In conclusion, NSER can be successfully performed in Dorper ewes by using a cervical relaxation protocol without oestradiol benzoate.

Preovulatory follicular dynamics, ovulatory response and embryo yield in Lacaune ewes subjected to synchronous estrus induction protocols and non-surgical embryo recovery.

The objective of this study was to assess the effect of the duration of progesterone-based estrus induction protocols on preovulatory follicular dynamics, ovulatory response, and embryo yield after non-surgical embryo recovery (NSER) in Lacaune ewes. Females received acetate medroxyprogesterone intravaginal sponges for six (G-6; n = 14) or nine (G-9; n = 14) days plus d-cloprostenol and eCG 24 h before sponge removal (Day 0). Preovulatory follicular dynamics and the luteal characteristics are evaluated by B-mode and Color-Doppler ultrasonography. NSER was performed five to six days after ovulation. The estrous behavior rate was 85.7% for both groups, and the percentage of ewes that ovulated was 92.9% in G-6 and 100% in G-9. The day of wave emergence (relative to Day 0) did not differ (P > 0.05) between G-6 (-3.0 ±â€¯0.5) and G-9 (-4.2 ±â€¯0.5). The number of follicles of size 4.1-5.0 mm was higher (P < 0.05) in G-9 (1.4 ±â€¯0.2) compared to G-6 (0.8 ±â€¯0.2) during the Days -4 to 0. At NSER, the transcervical penetration rate was 95.2% (20/21) and its duration time was lower (P < 0.05) in G-9 (3.4 ±â€¯0.6 min) than in G-6 (7.2 ±â€¯1.3 min). The number of ovulations and viable embryos was higher (P < 0.05) in G-9 (2.9 ±â€¯0.3 and 1.3 ±â€¯0.4, respectively) than in G-6 (1.9 ±â€¯0.3 and 0.4 ±â€¯0.2, respectively). In conclusion, the 9-day protocol promoted higher ovulation rate and embryo yield; moreover, the cervical dilation treatment allowed NSER in a high percentage of Lacaune ewes.

Recent advances in superovulation in sheep / Recentes avanços na superovulação de ovelhas

Nearly three years ago our research team published a review of intrinsic determinants and predictors of superovulatory responses in ewes (18th International Congress on Animal Reproduction, Tours, France; 26-30 June 2016; Theriogenology 2016;86:130-143). Heres a summary of major advances in superovulatory treatments of ewes reported over the last three years and of some most anticipated future directions. The rate of genetic improvement within a synthetic population of animals is significantly enhanced by an application of superovulation and embryo transfer. However, the sheep industry has progressed rather slowly in terms of implementing these technologies in comparison to other species although sheep have been used in many studies that paved the way to the development of modern assisted reproductive technologies (ARTs). Recently, several improvements in determining ovarian responses and non-invasively collecting/transferring embryos have been reported but some most frustrating problems associated with hormonal ovarian superstimulation (e.g., unpredictable and highly variable individual responses) remain unresolved. This warrants continued studies of both the physiological basis of and novel approaches to superovulatory treatments in sheep.(AU)

Recent advances in superovulation in sheep / Recentes avanços na superovulação de ovelhas

Nearly three years ago our research team published a review of intrinsic determinants and predictors of superovulatory responses in ewes (18th International Congress on Animal Reproduction, Tours, France; 26-30 June 2016; Theriogenology 2016;86:130-143). Here’s a summary of major advances in superovulatory treatments of ewes reported over the last three years and of some most anticipated future directions. The rate of genetic improvement within a synthetic population of animals is significantly enhanced by an application of superovulation and embryo transfer. However, the sheep industry has progressed rather slowly in terms of implementing these technologies in comparison to other species although sheep have been used in many studies that paved the way to the development of modern assisted reproductive technologies (ARTs). Recently, several improvements in determining ovarian responses and non-invasively collecting/transferring embryos have been reported but some most frustrating problems associated with hormonal ovarian superstimulation (e.g., unpredictable and highly variable individual responses) remain unresolved. This warrants continued studies of both the physiological basis of and novel approaches to superovulatory treatments in sheep.

The effect of mare's age on multiple ovulation rate, embryo recovery, post-transfer pregnancy rate, and interovulatory interval in a commercial embryo transfer program in Argentina.

Advanced maternal age is an important predisposing factor on the reduction of reproductive efficiency. The aim of this study was to evaluate the effect of donor's age on several reproductive parameters in a commercial equine embryo transfer program. Donors were classified into 3 age groups: Group 1=fillies (3 and 4 years old), Group 2=middle age mares (aged 5-10) and Group 3=old mares (aged 13-25). Embryo recovery, multiple ovulation and pregnancy rates and interovulatory intervals were compared amongst age groups. Group 1 (171/244, 70.1%) and Group 2 (774/1081, 71.6%) had a higher (P<0.005) embryo recovery rate than Group 3 (385/701, 54.9%). Groups 2 and 3 were 2.5 and 3.4 times more likely to have multiple ovulations than Group 1 (P<0.05), respectively. The effect of age group on pregnancy rate was not significant (P>0.05). The interovulatory intervals length was influenced by individual mare (P<0.001), age (P<0.04), Day of flushing (P=0.009) and by month (P<0.012). The overall mean interovulatory interval of Group 1 (16.4±0.17 days) and Group 2 (16.6±0.12 days) was not different (P>0.05), but was shorter than the one of Group 3 (17.4±0.15 days; P<0.04). The embryo recovery rate of flushings from Groups 1 and 2 was influenced by the length of the previous interovulatory interval (P=0.03).

Uso de procedimentos não cirúrgicos para a produção, recuperação e inovulação de embriões em pequenos ruminantes / In vivo embryo production, recovery and transfer in small ruminants by non surgical techniques

O objetivo deste artigo foi descrever técnicas não cirúrgicas associadas à recuperação e inovulação de embriões em caprinos e ovinos. Os procedimentos relativos a estas etapas da produção in vivo de embriões destes animais foram descritos de forma sequencial e detalhada, a fim de permitir sua execução por veterinários especialistas na área.(AU)

The objective of this article was to describe non surgical techniques related to recovery and transfer of sheep and goat embryos. The main procedures for these steps in the in vivo embryo production from these animals were described sequentially and with the requested details to support their performance by veterinarian experts.(AU)

Uso de procedimentos não cirúrgicos para a produção, recuperação e inovulação de embriões em pequenos ruminantes / In vivo embryo production, recovery and transfer in small ruminants by non surgical techniques

O objetivo deste artigo foi descrever técnicas não cirúrgicas associadas à recuperação e inovulação de embriões em caprinos e ovinos. Os procedimentos relativos a estas etapas da produção in vivo de embriões destes animais foram descritos de forma sequencial e detalhada, a fim de permitir sua execução por veterinários especialistas na área.

The objective of this article was to describe non surgical techniques related to recovery and transfer of sheep and goat embryos. The main procedures for these steps in the in vivo embryo production from these animals were described sequentially and with the requested details to support their performance by veterinarian experts.