Candida Genus Maximum Incidence in Boar Semen Even after Preservation, Is It Not a Risk for AI though?

There is little information in the literature about the fungal contamination of boar semen and its persistence during storage. The challenge of this study was to perform a mycological screening to identify the yeast in the raw semen at 12/24 h after dilution. The research was done in pig farms in the N-E area of Romania, with maximum biosecurity and state-of-the-art technology. All the examined ejaculates (101) were considered to be normal for each spermogram parameter, with microbiological determinations in T0 at the time of ejaculate collection, T1 at the time of dilution, and T2 at 24 h of storage. Microbiological determinations (mycological spermogram) were performed for quantitative (LogCFU/mL) and qualitative (typification of fungal genera) identification. Bacterial burden (×103 LogCFU/mL) after dilution (T1) decreased drastically (p < 0.0001) compared to the one in the raw semen (T0). After 24 h of storage at 17 °C, the mean value of the bacteriospermia remained constant at an average value of 0.44. Mycospermia had a constant trend at T0 (raw) and T1 (0.149 vs. 0.140) and was slightly higher at T2 (0.236). The difference between T1 vs. T2 (p = 0.0419) was close to the statistical reference value (p = 0.05). Of the total genera identified (24), the fungi had a proportion of 37.4% (9/15) and a ratio of 1:1.6. Regarding the total species (34), the fungi had a frequency of 29.42% (10/24) with a ratio between the fungi and bacteria of 1:2.4. A fertility rate of 86% was observed in the L1 group (50 AI sows with doses and mycospermia from T1), and an 82% rate was observed in the L2 group (50 AI sows with doses and mycospermia from T2). The litter size of L1 was 9.63 piglets and 9.56 for L2. Regarding the total number of piglets obtained between the two groups, there was a slight decrease of 22 piglets in group L2, without statistical differences (p > 0.05). The predominant genera persisted after dilution during a 12 h storage at 17 °C, where yeasts, such as Candida parapsilosis and C. sake were identified in more than 92% of AI doses.

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.

Total Aseptization of Boar Semen, to Increase the Biosecurity of Reproduction in Swine.

The aim of the study was to establish the complete microbiological profile of boar semen (Sus scrofa domesticus) and to choose the most effective antiseptic measures in order to control and optimize AI reproduction in pig farms. One hundred and one semen samples were collected and analyzed from several pig farms. The microbiological profile of ejaculates was determined by evaluating the degree of contamination of fresh semen and after dilution with specific extenders. The bacterial and fungal load of fresh boar semen recorded an average value of 82.41/0.149 × 103 CFU/mL, while after diluting the ejaculates the contamination value was 0.354/0.140 × 103 CFU/mL. Twenty-four germs (15 bacterial and 9 fungal species) were isolated, the most common being Candida parapsilosis/sake (92%) and Escherichia coli (81.2%). Modification of the sperm collection protocol (HPBC) reduced contamination in raw sperm by 49.85% in bacteria (significant (p < 0.00001) and by 9.67% in fungi (non-significant (p < 0.111491). The load in bacteria and filamentous fungi can be controllable, but not in levuras fungi. Some fluconazole-added extenders (12.5 mg%), ensure fungal aseptization, and even an increase in sperm progressivity (8.39%) for at least a 12 h shelf life after dilution. Validation of sperm aseptization was done by maintaining sow fecundity unchanged after AI (insignificant p > 0.05).

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.