Upgrading the fixed-time artificial insemination (FTAI) protocol in Romanian buffaloes.

The present study describes the challenges of assisted reproduction in Romanian buffaloes while increasing the efficacy of artificial insemination by choosing the most suitable method. The modified fixed-time artificial insemination (FTAI) protocol with sexed semen was used to increase the conception rate. This study included a total of 80 buffalo heifers that received ovarian stimulation using the OvSynch protocol. Two groups (n = 40), namely, a control group, in which the classic FTAI method was performed, and an experimental group, in which deep intrauterine AI was performed in cows that had developed a dominant follicle (US+UcFTAI), were randomly selected. The conception rate (CR) was 63.6% in the experimental group, which was statistically higher (P < 0.05) than the control group (30%). The ultrasound examination indicated that, using the OvSynch protocol, 82.5% (33 out of 40) of buffaloes developed a dominant follicle (DF) while the distribution between the warm and cold seasons was 75 and 90%, respectively. The CR was 60% during the hot season and 66.6% during the cold season. At calving, 92.5% female fetuses were born. The improved FTAI method in this study enhanced the results by reducing the waste of sexed semen and maximizing the response to OvSynch, making it a recommendation for practitioners. This study presents preliminary results and highlights that genetic progress is difficult to achieve. A systematic approach is needed in order to choose the most suitable biotechnological method for each farm.

Ovarian response to P4-PGF-FSH treatment in Suffolk sheep and P4-PGF-PMSG synchronization in cross-bred ewes, for IVD and ET protocol.

BACKGROUND: The success of an embryo transfer protocol in sheep depends on many factors, but the choice of drugs for the desired superovulation as well as the conception rate (CR) are most essential. Reproductive activity in sheep is characterized by a seasonality influenced by several factors such as photoperiod, latitude, temperature, nutrition and breed. Reproductive seasonality and nutritional condition are the main factors that influence embryo production in sheep. In sheep, some anatomical peculiarities limit the application of traditional reproductive biotechnologies used in cattle. OBJECTIVES: The aim of this study was to conclude on the effectiveness of a wider on farm in vivo embryo transfer development programme in Suffolk sheep by streamlining hormone therapies and optimizing technique. METHODS: A total number of 60 sheep and three rams were included in this study, divided into two groups (receptors and donors). Donor Suffolk sheep were treated for superovulation using the P4-PGF-FSH multiple ovulation embryo transfer (MOET) protocol, while the cross-bred recipients' group was synchronized with P4-PGF-PMSG. RESULTS: On the first day after superovulation, all ovaries had more than five dominant follicles, while corpora lutea were later observed in 83.3% sheep. The recovery rate was 83.3%, while 72.9% embryos were transferable. Embryos were transferred directly into recipients. Fertility after 30 days was 68.57%, lambing rate was 91.6% and CR was 62.85%. This study showed that veterinary drugs (P4, FSH, LH, PMSG, PGF) used for superovulation optimized by us were capable of producing by this improved technique the optimization of the reproduction indices at embryo-transfer (ET) and to be able to be used successfully. CONCLUSIONS: The application of an MOET protocol has a positive effect in the production of in vivo embryo production (IVD) embryos in Suffolk sheep and can guarantee the success of embryo transfer activity to ewes with lower genetic merit. Our research aimed at representing a model for sheep farms for a rapid improvement of productive traits.

Differences in CASA output according to the chamber type when analyzing frozen-thawed bull sperm.

As demonstrated by some authors, the type of analyzing chamber can greatly influence the results of computer-assisted sperm analysis (CASA). This study aimed to compare three of the disposable chamber types currently available on the market and to determine whether the CASA output may be significantly different among them. The semen from five Fleckvieh bulls was analyzed by CASA using three different disposable chambers: Leja (20µm), MofA (20µm) and Minitube (20µm), at three different time points: immediately after filling the chamber, at 6min, and also at 12min after filling. Sperm concentration was also determined using the Nucleocounter® NC-100™ device and the hemocytometer as standard methods. The results showed higher values in terms of total and progressive sperm motility for MofA compared to the other two chambers immediately after filling (p<0.05), but higher values for Leja and Minitube after 6 and 12min (p<0.05). All three disposable chambers offered lower values for sperm concentration compared to standard methods (Leja: 68.4±4.9×106/mL; MofA: 80.8±9.6×106/mL; Minitube: 67.3±5.4×106/mL; Nucleocounter: 86.5×106/mL; Hemocytometer: 84.0×106/mL). We conclude that for rapid analyses the MofA chambers provide superior results when compared to the other types that we tested. However, when the analysis requires a longer duration, the Minitube type, and especially the Leja type provide a greater degree of confidence. Further, for determining sperm concentration we think that examiners would be more accurate using the Nucleocounter or the hemocytometer and should make use of CASA only when the other methods are not available.