In vitro effects of lipopolysaccharides on bovine uterine contractility.

Metritis is an important disorder in dairy cows during the early postpartum period. Myometrial contractility is a prerequisite for uterine involution; however, very scanty literature is available about the effect of metritis on this process and endocrine responsiveness. This study was aimed to evaluate the effect of inflammation on uterine contractility in vitro, and the inflammation was induced by incubating myometrial strips with lipopolysaccharides (LPS). Myometrial samples were collected from 17 healthy Holstein Friesian cows during caesarean section. Eight longitudinal strips from each cow were incubated in organ baths with LPS concentrations of 0 (LPS0 ), 0.1 (LPS0.1 ), 1 (LPS1 ) and 10 µg/ml (LPS10 ). Spontaneous contractility and contractility induced by increasing concentrations of oxytocin (10-10 - 10-7  mol/L) were recorded during nine 30-min intervals (T1 to T9). The minimum amplitude (minA), maximum amplitude (maxA), mean amplitude (meanA) and area under the curve (AUC) were calculated for each time interval. LPS had an effect (p ≤ .05) on maxA, meanA and AUC. In T1, myometrial strips incubated with LPS0.1 and LPS1 had higher (p ≤ .05) maxA, meanA and AUC than the strips incubated with LPS0 . In T9 without oxytocin, LPS0 led to higher (p ≤ .05) maxA, meanA and AUC than LPS0.1 and LPS1 . In T8 and T9 with oxytocin, LPS1 had lower (p ≤ .05) maxA, meanA and AUC than the other LPS concentrations. Interestingly, the results show that LPS has a transient positive effect on myometrial contractility in vitro and that this effect is dependent on LPS concentration and duration of incubation.

[Gender determination in equine fetuses in early pregnancy using two- and three-dimensional ultrasound]. / Geschlechtsbestimmung bei Pferdefeten mittels zwei- und dreidimensionaler Sonografie in der Frühgravidität.

AIM: The aim of this study was to compare transrectal two-dimensional (2D) and three-dimensional (3D) ultrasound examination with regards to required time and accuracy of fetal sex determination in early pregnant mares. MATERIALS AND METHODS: For this purpose 47 mares were examined transrectally once between days 58 and 115 of gestation. Initially, the fetal sex was determined by identifying the location of the genital tubercle (GT) or external genitalia using 2D-ultrasound. Subsequently, the ultrasound machine was switched to 3D-mode to obtain images for later computer-based evaluation. RESULTS: The gestational period between days 58 and 79 of pregnancy was the most appropriate time for sex determination with 77 % (2D and first 3D evaluation) correct diagnoses. The accuracy of the sex determination could be increased by about 16 % by a second evaluation of the 3D-images in a minimum time interval of 2 weeks. For each mare the additional time needed to perform the 3D-examination and to assess the 3D-images was approximately 6-7 minutes. CONCLUSION AND CLINICAL RELEVANCE: This study demonstrates that the accuracy of transrectal fetal gender determination is higher by using 3D-mode compared to the 2D-ultrasound. For experienced examiners however, 3D ultrasound technology does not offer any decisive advantages and is also more time-consuming.

Effects of Different Freezing Protocols on Motility, Viability, Mitochondrial Membrane Potential, Intracellular Calcium Level, and DNA Integrity of Cryopreserved Equine Epididymal Sperm.

The aim of the present study was to evaluate the effect of different freezing procedures on sperm motion, viability, the acrosome status, mitochondrial membrane potential (MMP), intracellular calcium content, and DNA integrity on epididymal stallion sperm. Therefore, the sperm of 10 healthy stallions was harvested by retrograde flushing after testectomy, diluted with a semen extender containing defined milk proteins and a freezing extender containing egg yolk and glycerol and frozen according to 4 different protocols, using a programmable freezer and a floating rack performing a slow (processes 1 and 2) or a fast cooling rate (processes 3 and 4, respectively). Post-thaw total motility and slow sperm values were lower when using process 4 compared with processes 1 and 2 (P < .05) after 1 hour of incubation. Progressive motility was lower in process 4 compared with process 1 immediately after thawing and after 1 hour of incubation (P < .05). The amount of rapid sperm was lower when using process 4 compared with process 1 immediately after thawing (P < .05). After 1 hour of incubation, the amount of rapid sperm was lower when using process 4 compared with processes 1 and 2 (P < .05). Higher values for viable sperm were seen in processes 1 and 2 compared with process 4 (P < .05) after 1 hour of incubation. Immediately after thawing, more viable sperm with high MMP (hMMP) were observed when using process 3 compared with process 2 (P < .05). After 1 hour of incubation, a significantly higher amount of viable hMMP sperm were detected when using processes 1 and 2 compared with process 4 (P < .05). Process 2 yielded a lower percentage of sperm containing low calcium (lCa) than process 3 immediately after thawing (P < .05). After 1 hour of incubation, the lowest amount of lCa sperm was observed using process 4 (P < .05). The subpopulation of viable/hMMP/lCa sperm was higher when using process 3 compared with process 2 immediately after thawing (P < .05). After 1 hour of incubation, the lowest amount of this subpopulation was detected in process 4 (P < .05). The DNA integrity was similar in all groups. In conclusion, a slow cooling rate with a controlled rate freezer resulted in best sperm quality after thawing. Using a floating rack in nitrogen vapor as an alternative to a programmable freezer, equilibration in a cooled environment is advantageous.

Herbal yeast product, Equi-Strath®, alters the antioxidant status of stallion semen.

Effects of a plasmolysed yeast product enriched with herbs, malt, honey and orange syrup on semen characteristics and oxidative status in stallions were evaluated. Twenty stallions (mean age ±â€¯standard deviation = 9.5 ±â€¯4.5 years) were randomly divided into a treatment group (n = 10) receiving 0.06 mL/kg bodyweight of plasmolysed herbal yeast, and a control group (n = 10) receiving the same amount of placebo daily in the feed for 10 weeks. Ejaculates were collected weekly from all stallions starting at Week 0. Volume, sperm concentration, motility, and velocity were evaluated immediately, 24 and 48 h after cooled storage at 5 °C. At the two storage time points, membrane lipid peroxidation was determined using the BODIPY-C11. Additionally, blood samples were collected at Weeks 0, 1, 5 and 9, and analysed for antioxidant status, consisting of superoxide dismutase, cholesterol, thiobarbituric acid reactive substances, and non-esterified fatty acids. Due to the nature of the data, the Mann-Whitney U test was applied as preliminary analysis. The BODIPY-C11 in the semen was less at 24 h and greater at 48 h after collections in Week 1 to 3 (P < 0.01) and Week 1 to 10 (P <  0.05) compared with Week 0 in the treatment compared to control group. There were no significant differences between groups for all values for other seminal and blood variables evaluated. In conclusion, feed supplementation with plasmolysed herbal yeast temporarily improved the antioxidant status of stallion semen, which might be of benefit for preservation of cooled semen.