100 COMPARISON OF PREGNANCY RATES OF FRESH IN VITRO-PRODUCED BOS INDICUS EMBRYOS PRODUCED IN THE SAME LABORATORY BUT COLLECTED AND TRANSFERRED IN PANAMA OR THE DOMINICAN REPUBLIC

2014 ◽  
Vol 26 (1) ◽  
pp. 164
Author(s):  
L. F. Nasser ◽  
S. C. Feliú ◽  
E. Rodríguez ◽  
K. Mojica ◽  
E. G. Oliveira ◽  
...  

Because of Panama's stricter sanitary status, a specialised protocol was developed with the Department of Agriculture in the Dominican Republic to legalize the exchange of biological materials (oocytes/embryos). This protocol allows the team of specialised technicians, currently working in Born® Animal Biotechnology's Panamanian facility, to operate using the same in vitro bovine embryo production system (IVP, In vitro Brasil®) to service Dominican producers. Because the donors are not located at a specific centre with controlled sanitary management, a special protocol was developed in which blood tests were done to certify that the entirety of the herd at each client's farm was free of infectious bovine rhinotracheitis, DBVD, leptospirosis, leucosis, brucellosis, and tuberculosis. As timing during IVP is an essential variable that can have detrimental effects on the final results, precautions were taken to ensure that the oocytes arrived at the Panamanian laboratory facility within 24 h of aspiration. A portable incubator was used to transport oocytes and embryos during the import and export portions of the procedure. A comparison of pregnancy rates based on oocyte source and recipient transfers from September 2012 until May 2013 was analysed with ?2 (Table 1). The number of embryos produced in Panama was significantly higher than in the Dominican Republic, which was likely due to the larger number of donors and oocytes from the Panama herd. However, pregnancy rate was higher in the Dominican Republic likely because of the health status of the Dominican recipients, which were free of the diseases mentioned above. Recipients were the same type and breed and under similar management conditions in both countries. The disease status aspect will be examined with greater numbers of animals in the future. The data suggest that the present IVP and recipient management protocols could serve as a model for other Central American and Caribbean countries under similar management systems. Table 1.In vitro embryo production and pregnancy rates of Bos indicus embryos transferred in Panama or the Dominican Republic (September 2012 through May 2013)

2012 ◽  
Vol 24 (1) ◽  
pp. 196
Author(s):  
M. P. Palhão ◽  
E. R. Oliveira ◽  
M. M. Gioso ◽  
B. C. Carvalho ◽  
L. G. B. Siqueira ◽  
...  

The ovarian follicular population has been used as a parameter to evaluate fertility and also the potential of donors undergoing assisted reproductive procedures in both human medicine and animal practice. There is a high correlation between follicular population and oocyte recovery by ovum pickup (OPU), but the relationship between oocyte recovery, embryo production and pregnancy rates may not be fully understood. The aim of the present study was to evaluate whether the conversion rate of oocytes to embryos and further pregnancies could be positively related to the number of cumulus–oocyte complexes (COC) recovered after OPU in cattle. For this purpose, records of 626 OPU sections from 251 nonlactating Gyr cows (dairy Zebu breed) were analysed. The animals had a good body condition score, were kept in a good feeding pasture (Brachiaria spp.) and were supplemented with corn silage and a mixture of corn, soybeans and vitamin and minerals, according to their nutritional requirements. For each ovarian aspiration, the ovarian follicular wave was previously synchronized with an auricular implant (Norgestomet-Crestar®), IM injections of 2 mg of oestradiol benzoate (Gonadiol®) and 0.25 mg of D-cloprostenol (Sincrocio®). The OPU procedures were performed using an ultrasound device (Aquila Vet, Esaote, São Paulo, Brazil) equipped with a vaginal sector 7.5-MHz probe, disposable 20 G needles and a vacuum pressure of 80 mmHg. The cows were ranked in quartiles regarding the total number of COC recovered. To reduce bias related to the eventual fluctuation of OPU results, for the present analysis the authors used only the recorded OPU session of each cow with the highest number of COC recovered. Viable COC were fertilized with sex-sorted (X) semen of Gyr bulls previously tested for in vitro embryo production. Conversion rates (%) of the total and viable oocytes to embryos, viable oocytes to pregnancy and embryo to pregnancy were evaluated for each quartile. Differences between the first and fourth quartiles were accessed by Fisher's exact test. In the 251 OPU, 4246 total and 3173 viable COC were recovered, resulting in the production of 1001 embryos (31.5%) and 453 pregnancies (45.3%). The cows ranked in the first, second, third and fourth quartiles produced >30 (41.6 ± 10.6), 21 to 30 (25.2 ± 3.0), 12 to 20 (15.9 ± 2.6) and <12 (6.7 ± 3.1) total oocytes. The average viable oocyte (29.1 ± 11.0, 18.1 ± 5.3, 11.1 ± 3.7 and 4.5 ± 2.7, respectively) and embryo production (8.6 ± 5.7, 5.2 ± 3.6, 3.8 ± 2.8 and 1.8 ± 1.8, respectively) were different (P < 0.0001) among all quartiles. Pregnancy rates, however, did not differ (46.0, 44.9, 43.9 and 45.6%, respectively; P > 0.05). Interestingly, the conversion rates (viable oocytes to embryos and viable oocytes to pregnancies) were higher (P < 0.0001 and 0.002) in cows from the last quartile (51.1 and 31.9%) compared with those from the first quartile (23.7 and 14.7%). In conclusion, the number of COC recovered by OPU (and consequently the ovarian follicular count) can further predict the total number of embryos and pregnancies produced, but it is not directly related to the oocyte development potential. Biotran and Fapemig Project CVZ APQ 01654/09 and BPD 0007/10.


2004 ◽  
Vol 16 (2) ◽  
pp. 212
Author(s):  
L.F. Nasser ◽  
E.L. Reis ◽  
A.M. Oliveira ◽  
G.A. Bo ◽  
P.S. Baruselli

It has been shown recently that treatments with progesterone (P4)-releasing devices combined with estradiol benzoate (EB) plus P4 on Day 0, eCG and PGF on Day 5 and a second application of EB one day after device removal (Day 9) can be used successfully to transfer bovine embryos at a self-appointed time, without the necessity of estrus detection. Although the treatment solved one of the major problems in recipient management, estrus detection, it requires handling the recipients at least five times for treatments and embryo transfer. An experiment was designed to evaluate whether reducing one day of handling, by the administration of eCG and PGF at the time of removal of the P4 device (Day 8), results in comparable pregnancy rates than giving eCG on Day 5. A secondary objective was to determine the effect of injectable P4 at the time of device insertion plus EB treatment. Crossbred Bos taurus×Bos indicus beef heifers (n=301) were randomly assigned to 4 treatment groups in a 2 by 2 factorial design. All Heifers received a P4 device (DIB, Syntex, Argentina) plus 2mg EB i.m. (Syntex) at unknown stages of the estrous cycle (Day 0), with or without 50mg of P4 given i.m. at the same time. Heifers were further subdivided to receive PGF (0.150mg d-cloprostenol, Prolise, Tecnopec, Sao Paulo, Brazil) and 400IU of eCG (Novormon, Syntex) i.m. on Days 5 or 8. In all heifers, DIB devices were removed on Day 8 and 1 mg EB was administered i.m. on Day 9. Day 10 was arbitrarily considered as the day of estrus. On Day 17, heifers were bled for plasma P4 concentrations and examined by ultrasonography to determine the number of CL and their diameter. Heifers that had &gt;1 CL or a single CL with diameter ≥18mm received an in vitro-produced (IVP) embryo by nonsurgical transfer performed by the same veterinarian. Pregnancy rates were determined by ultrasonography 30 days later. The effects of Day of eCG administration (Day 5 or Day 8), P4 of treatment (E2 or E2+P4) and the day-by-P4 treatment interaction on the numbers of CL and plasma P4 were analyzed by ANOVA, and the proportion of recipients selected and pregnant were analyzed using non-parametric tests (NPAR1WAY, SAS). There was no significant effect of P4 treatment or the P4-by-day of eCG interaction in any of the parameters evaluated. However, there was a significant effect of day of eCG administration on plasma P4 concentrations (Day 5=2.4±0.3 v. Day 8=1.7±0.2; P=0.03) and the number of CL (Day 5=1.4±0.1 v. Day 8=1.1±0.0; P=0.02) on Day 17. Furthermore, the proportion of recipients pregnant/treated tended (P=0.1) to be higher in heifers in the Day 5 Group (71/151, 47.0%) than in those in the Day 8 Group (61/150, 40.7%). Although delaying the eCG and PGF administration from Day 5 to Day 8 saves one trip through the chute for treatments, it resulted in lower plasma P4 concentrations and tended to decrease pregnancy rates in bovine embryo recipients synchronized with DIB devices and EB and transferred at a fixed time. Furthermore, the administration of injectable P4 at the time of DIB insertion did not affect pregnancy rates.


2015 ◽  
Vol 27 (1) ◽  
pp. 210
Author(s):  
L. F. Feres ◽  
L. S. A. Camargo ◽  
M. P. Palhao ◽  
F. Z. Brandao ◽  
J. H. M. Viana

Improving in vitro culture systems to optimize embryo yield has been a major research goal. The relationship between the efficiency of embryo production systems and the pregnancy outcomes, however, remain controversial. The aim of the present study was to evaluate the likelihood of pregnancy of in vitro-produced embryos derived from batches with different relative efficiency indexes. Data of 702 ovum pick-up (OPU) and in vitro embryo production (IVEP) sessions, and of 2456 embryo transfers, recorded from 2008 to 2012, were evaluated. All donors were from the same herd, and were of the same breed (Gir, Bos indicus), as well as the semen used for IVF. The cumulus-oocycte complex (COC) recovery and IVEP were performed by the same team, in a single IVF laboratory, and using standard medium and procedures. Only data from embryos transferred as fresh were used, and records from 97 OPU/IVEP sessions in which no embryo was produced, or embryos were frozen or discharged due to lack of recipients, were discharged. The remaining 605 sessions were stratified in quartiles (I to IV, each one corresponding to 25% of total data) according to COC production of the donors, or stratified in ranges (0–25%, 26–50%, 51–75%, and 76–100%) according to COC quality (percentage of viable COC or of grade I COC) and to embryo production efficiency endpoints (cleavage rate, blastocyst rate). Pregnancy rates were compared among quartiles or ranges by the chi-square method. On average, the Gir donors produced 24.8 ± 0.6 COC per OPU, from which 14.4 ± 0.4 were classified as viable (57.8%), and 3.2 ± 0.1 as grade I (12.9%). On average 6.1 ± 0.2 embryos (morulas and blastocysts) were produced per OPU per donor, and mean pregnancy rate was 30.9%. As expected, donors with greater total COC yield (quartile I) also produced more viable oocytes (25.5 ± 0.7 v. 15.7 ± 0.3, 10.5 ± 0.2 and 5.8 ± 0.2), more COC grade I (4.8 ± 0.4 v. 3.9 ± 0.3, 2.6 ± 0.2 and 1.6 ± 0.1), and more embryos (9.0 ± 0.4 v. 6.9 ± 0.3, 5.0 ± 0.2 and 3.3 ± 0.1) than donors from quartiles II, III, or IV, respectively (P < 0.0001). Nevertheless, there was no difference (P > 0.05) in pregnancy rates for embryos produced from donors ranked in the different quartiles (30.9 v. 29.3, 31.5, and 30.5% for quartiles I to IV, respectively). Similarly, there was no difference (P > 0.05) in the pregnancy rate of embryos derived from OPU sessions in which there was a high or low percentage of viable or grade I COC. In vitro production efficiency (cleavage and blastocyst rates) also had no effect (P > 0.05) on further pregnancy rates. In conclusion, these results suggest that there is no relationship among the average number or quality of the COC recovered by OPU, the efficiency of IVEP, and the likelihood of pregnancy of in vitro-derived embryos.Research was supported by Fazendas do Basa, CNPq, and Fapemig.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 37-37
Author(s):  
Nicola Oosthuizen ◽  
Pedro Levy Piza Fontes ◽  
G Cliff Lamb

Abstract Bos indicus and Bos taurus cattle diverged from an evolutionary standpoint more than 110,000 years ago. Since then, Bos indicus cattle have undergone genetic adaptations beyond the commonly discussed increased thermo-tolerance and parasite resistance. Several physiological differences exist between Bos indicus and Bos taurus cattle, and it is important to consider these differences when establishing reproductive management strategies. It has been well described that Bos indicus cattle have a delayed onset of puberty and longer periods of postpartum anestrus, yet through the utilization of estrus synchronization protocols these challenges can be attenuated. However, when Bos indicus females are exposed to these protocols, they are known to have smaller dominant follicles, lower expression of estrus, and decreased pregnancy rates to artificial insemination (AI) when compared to Bos taurus females. These factors can be overcome through the utilization of estradiol and progesterone based synchronization protocols, which improve follicular dynamics and yield acceptable pregnancy rates to assisted reproductive technologies in cattle adapted to tropical or subtropical conditions. However, the use of estrogens for synchronization purposes is not permitted in the United States, and cattle producers need to rely on GnRH-based protocols. Another key difference between subspecies, is that Bos indicus females have greater antral follicle counts than Bos taurus females, which proves beneficial for in vitro embryo production. Therefore, an opportunity exists to explore the greater productivity of donors in embryo production in order to improve genetics in herds that utilize these breeds.


2011 ◽  
Vol 75 (9) ◽  
pp. 1640-1646 ◽  
Author(s):  
J.H.F. Pontes ◽  
F.A. Melo Sterza ◽  
A.C. Basso ◽  
C.R. Ferreira ◽  
B.V. Sanches ◽  
...  

2014 ◽  
Vol 26 (1) ◽  
pp. 185
Author(s):  
A. Nagele ◽  
E. Gomes ◽  
A. Ruiz ◽  
L. F. Nasser ◽  
S. Feliu ◽  
...  

It has been previously demonstrated (IETS 2011) that Panama is applying the biotechnology of in vitro embryo production (IVP) to their bovine reproduction management systems. The present work demonstrates the ability to apply the IVP technology across 2 distant country borders. Herein, we demonstrate that a country (Dominican Republic; DR) that does not have a bovine IVP laboratory can take advantage of fresh bovine IVP embryos for transfer using distant IVP facilities in another country (Panama; ~1500 km away). The objective of this study was to demonstrate that a model system for large-scale commercial in vitro bovine embryo production for beef and dairy producers, that do not have IVP technology in their home country, could be developed producing comparable results. As the same laboratory provides IVP services to the both countries, a special sanitary protocol was developed in order to legalize the exchange of biological materials (oocytes or embryos). The data obtained in DR was compared to Panamanian client data because identical conditions were utilised for IVP. Cattle production systems were similar, as Brahman (a Zebu type of cattle) is the most popular breed in both countries. Oocytes were collected from 10 different herds in Panama and 4 different herds in DR. The oocytes were transported in an oocyte transporter in both instances. However, oocytes from DR were transported in InVitro Brasil™ maturation medium from 12 to 18 h and in Panama from 6 to 12 h before they were placed in a standard CO2 incubator. In both cases, the oocytes were matured for 24 h before fertilization with conventionally frozen Brahman semen in InVitro Brasil™ fertilization medium, followed by culture for up to 7 days in InVitro Brasil™ embryo culture medium. The embryos were transferred on Day 7, either in Panama or DR. They were transported by car in Panama and via airplane back to DR. A comparison of oocyte number and quality, cleavage, embryo production, and pregnancy rate, was made using the same in vitro production system for Brahman donors from September 2012 until May 2013. The difference between sites in the relative number of viable oocytes, relative number of cleaved oocytes among viable oocytes, relative number of embryos produced among cleaved oocytes, and relative number of embryos produced among viable oocytes was tested using Fisher's exact test. Pregnancy rate was analysed with chi-squared. We realise these results represent field data; however, we believe the present work is a significant step in demonstrating the potential for wide commercial-scale dissemination of IVP technology between distant countries. The number of embryos produced in Panama was slightly, but significantly, higher than those produced in DR; this is likely due to the larger number of donors and oocytes from the Panamanian herds. However, the pregnancy rate was higher in DR, likely due to the health status of DR recipients. These data illustrate that IVP using Brahman donors could be used as a tool to improve and spread superior genetics. Furthermore, this technique can serve as a model for other Central American and Caribbean countries under similar management systems. Table 1.Panama and the Dominican Republic in vitro Brahman embryo production and pregnancy (September 2012 through May 2013)


2010 ◽  
Vol 74 (8) ◽  
pp. 1349-1355 ◽  
Author(s):  
J.H.F. Pontes ◽  
K.C.F. Silva ◽  
A.C. Basso ◽  
A.G. Rigo ◽  
C.R. Ferreira ◽  
...  

Zygote ◽  
2016 ◽  
Vol 24 (6) ◽  
pp. 890-899 ◽  
Author(s):  
A.L.S. Guimarães ◽  
S.A. Pereira ◽  
M. N. Diógenes ◽  
M.A.N. Dode

SummaryThe aim of this study was to evaluate the effect of adding a combination of insulin, transferrin and selenium (ITS) and l-ascorbic acid (AA) during in vitro maturation (IVM) and in vitro culture (IVC) on in vitro embryo production. To verify the effect of the supplements, cleavage and blastocyst rates, embryo size and total cell number were performed. Embryonic development data, embryo size categorization and kinetics of maturation were analyzed by chi-squared test, while the total cell number was analyzed by a Kruskal–Wallis test (P < 0.05). When ITS was present during IVM, IVC or the entire culture, all treatments had a cleavage and blastocyst rates and embryo quality, similar to those of the control group (P < 0.05). Supplementation of IVM medium with ITS and AA for 12 h or 24 h showed that the last 12 h increased embryo production (51.6%; n = 220) on D7 compared with the control (39.5%; n = 213). However, no improvement was observed in blastocyst rate when less competent oocytes, obtained from 1–3 mm follicles, were exposed to ITS + AA for the last 12 h of IVM, with a blastocyst rate of 14.9% (n = 47) compared with 61.0% (n = 141) in the control group. The results suggest that the addition of ITS alone did not affect embryo production; however, when combined with AA in the last 12 h of maturation, there was improvement in the quantity and quality of embryos produced. Furthermore, the use of ITS and AA during IVM did not improve the competence of oocytes obtained from small follicles.


1997 ◽  
Vol 47 (1) ◽  
pp. 265 ◽  
Author(s):  
E. Behboodi ◽  
A. Gutiérrez-Adán ◽  
G.B. Anderson

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